Identification of T. gondii myosin light chain-1 as a direct target of TachypleginA-2, a small-molecule inhibitor of parasite motility and invasion

This work was supported by US Public Health Service grant AI054961 (GEW/NJW), a University Research Fellowship from the Royal Society (NJW) and funding for the mass spectrometry analysis was provided by the Vermont Genetics Network/NIH Grant 8P20GM103449 from the INBRE program of the NIGMS.Motility of the protozoan parasite Toxoplasma gondii plays an important role in the parasite's life cycle and virulence within animal and human hosts. Motility is driven by a myosin motor complex that is highly conserved across the Phylum Apicomplexa. Two key components of this complex are the class XIV unconventional myosin, TgMyoA, and its associated light chain, TgMLC1. We previously showed that treatment of parasites with a small-molecule inhibitor of T. gondii invasion and motility, tachypleginA, induces an electrophoretic mobility shift of TgMLC1 that is associated with decreased myosin motor activity. However, the direct target(s) of tachypleginA and the molecular basis of the compound-induced TgMLC1 modification were unknown. We show here by ''click'' chemistry labelling that TgMLC1 is a direct and covalent target of an alkyne-derivatized analogue of tachypleginA. We also show that this analogue can covalently bind to model thiol substrates. The electrophoretic mobility shift induced by another structural analogue, tachypleginA-2, was associated with the formation of a 225.118 Da adduct on S57 and/or C58, and treatment with deuterated tachypleginA-2 confirmed that the adduct was derived from the compound itself. Recombinant TgMLC1 containing a C58S mutation (but not S57A) was refractory to click labelling and no longer exhibited a mobility shift in response to compound treatment, identifying C58 as the site of compound binding on TgMLC1. Finally, a knock-in parasite line expressing the C58S mutation showed decreased sensitivity to compound treatment in a quantitative 3D motility assay. These data strongly support a model in which tachypleginA and its analogues inhibit the motility of T. gondii by binding directly and covalently to C58 of TgMLC1, thereby causing a decrease in the activity of the parasite's myosin motor. Publisher PDFPeer reviewe

Measurement of the 1/E Dependence of the 7-Li(p,n)7-Be Total Reaction Cross Section

This work was supported by the National Science Foundation Grant NSF PHY 78-22774 A02 & A03 and by Indiana Universit

Cornerstones of Sampling of Operator Theory

This paper reviews some results on the identifiability of classes of operators whose Kohn-Nirenberg symbols are band-limited (called band-limited operators), which we refer to as sampling of operators. We trace the motivation and history of the subject back to the original work of the third-named author in the late 1950s and early 1960s, and to the innovations in spread-spectrum communications that preceded that work. We give a brief overview of the NOMAC (Noise Modulation and Correlation) and Rake receivers, which were early implementations of spread-spectrum multi-path wireless communication systems. We examine in detail the original proof of the third-named author characterizing identifiability of channels in terms of the maximum time and Doppler spread of the channel, and do the same for the subsequent generalization of that work by Bello. The mathematical limitations inherent in the proofs of Bello and the third author are removed by using mathematical tools unavailable at the time. We survey more recent advances in sampling of operators and discuss the implications of the use of periodically-weighted delta-trains as identifiers for operator classes that satisfy Bello's criterion for identifiability, leading to new insights into the theory of finite-dimensional Gabor systems. We present novel results on operator sampling in higher dimensions, and review implications and generalizations of the results to stochastic operators, MIMO systems, and operators with unknown spreading domains

Limaria hians (Mollusca : Limacea): a neglected reef-forming keystone species

1. A key component of physical habitat along braided river systems is the exposed riverine sediment within the active zone. The relatively unmanaged, gravel-bed Fiume Tagliamento, Italy, provides the focus for exploring two ecologically important properties of exposed riverine sediments: their within-patch and between-patch variability in calibre. 2. To characterize between-patch variation in exposed riverine sediments, replicate (within-patch) samples were obtained from three geomorphologically distinct locations along 130 km of the river: bar heads along the margin of the low-flow channel, the heads of major bars across the exposed surface of the active zone, and floodplain surfaces. A photographic technique enabled rapid and consistent field sampling of the coarse sediments at bar heads along the low-flow channel margin and on major bars across the dry bed. 3. A downstream decrease in particle size and an increase in within-patch heterogeneity in sediment size were observed within bar head sediments along the margin of the low-flow channel. Comparisons between major bar and low-flow channel samples revealed greatest within-patch variability in individual sediment size indices (D50, A- and B-axes of the larger particles) at headwater sites, greatest between-patch variability in the three measured indices in the central reaches, and lowest between-patch variability at downstream sites. However, there was a distinct increase in the overall heterogeneity in particle size, which was sustained across all patches, in a downstream direction. 4. There was a clear downstream decrease in the size of floodplain sediments in the headwaters, but thereafter there was no distinct downstream trend in any of the calculated particle size indices. 5. The geomorphological controls on the observed patterns and the potential ecological significance of the patterns, particularly for plant establishment, are discussed in relation to the relative relief of the active zone, and the highly variable hydrological and climatic regime along the river

The presence of GC-AG introns in Neurospora crassa and other euascomycetes determined from analyses of complete genomes: implications for automated gene prediction

AbstractA combination of experimental and computational approaches was employed to identify introns with noncanonical GC-AG splice sites (GC-AG introns) within euascomycete genomes. Evaluation of 2335 cDNA-confirmed introns from Neurospora crassa revealed 27 such introns (1.2%). A similar frequency (1.0%) of GC-AG introns was identified in Fusarium graminearum, in which 3 of 292 cDNA-confirmed introns contained GC-AG splice sites. Computational analyses of the N. crassa genome using a GC-AG intron consensus sequence identified an additional 20 probable GC-AG introns in this fungus. For 8 of the 47 GC-AG introns identified in N. crassa a GC donor site is also present in a homolog from Magnaporthe grisea, F. graminearum, or Aspergillus nidulans. In most cases, however, homologs in these fungi contain a GT-AG intron or no intron at the corresponding position. These findings have important implications for fungal genome annotation, as the automated annotations of euascomycete genomes incorrectly identified intron boundaries for all of the confirmed and probable GC-AG introns reported here

Application of Pulsed Field Gel Electrophoresis to Determine γ-ray-induced Double-strand Breaks in Yeast Chromosomal Molecules

The frequency of DNA double-strand breaks (dsb) was determined in yeast cells exposed to γ-rays under anoxic conditions. Genomic DNA of treated cells was separated by pulsed field gel electrophoresis, and two different approaches for the evaluation of the gels were employed: (1) The DNA mass distribution profile obtained by electrophoresis was compared to computed profiles, and the number of DSB per unit length was then derived in terms of a fitting procedure; (2) hybridization of selected chromosomes was performed, and a comparison of the hybridization signals in treated and untreated samples was then used to derive the frequency of dsb

Biological anthropology in the Indo-Pacific Region: New approaches to age-old questions

Biological anthropological research, the study of both modern and past humans, is a burgeoning field in the Indo-Pacific region. It is becoming increasingly apparent that the unique environments of the Indo-Pacific have resulted in an archaeological record that does not necessarily align with those in the northern hemisphere. New, regionally-specific archaeological models are being developed, and biological anthropological research has an important role to play in establishing past human experience within these models. In the Indo-Pacific, research using ancient and modern human tissues is adding insight into global processes of prehistoric settlement and migrations, subsistence change and human biosocial adaptation. This review synthesises current themes in biological anthropology in this region. It highlights the diverse methods and approaches used by biological anthropologists to address globally-relevant archaeological questions. In recent decades a collaborative approach between archaeologists, biological anthropologists and local communities has become the norm in the region. The many positive outcomes of this multi-disciplinary approach are highlighted here through the use of regionally-specific case studies. This review ultimately aims to stimulate further collaborations between archaeologists, biological anthropologists and the communities in the region, and demonstrate how the evidence from Indo-Pacific research may be relevant to global archaeological models

Long-term evolution of orbits about a precessing oblate planet. 3. A semianalytical and a purely numerical approach

Construction of a theory of orbits about a precessing oblate planet, in terms of osculating elements defined in a frame of the equator of date, was started in Efroimsky and Goldreich (2004) and Efroimsky (2005, 2006). We now combine that analytical machinery with numerics. The resulting semianalytical theory is then applied to Deimos over long time scales. In parallel, we carry out a purely numerical integration in an inertial Cartesian frame. The results agree to within a small margin, for over 10 Myr, demonstrating the applicability of our semianalytical model over long timescales. This will enable us to employ it at the further steps of the project, enriching the model with the tides, the pull of the Sun, and the planet's triaxiality. Another goal of our work was to check if the equinoctial precession predicted for a rigid Mars could have been sufficient to repel the orbits away from the equator. We show that for low initial inclinations, the orbit inclination reckoned from the precessing equator of date is subject only to small variations. This is an extension, to non-uniform precession given by the Colombo model, of an old result obtained by Goldreich (1965) for the case of uniform precession and a low initial inclination. However, near-polar initial inclinations may exhibit considerable variations for up to +/- 10 deg in magnitude. Nevertheless, the analysis confirms that an oblate planet can, indeed, afford large variations of the equinoctial precession over hundreds of millions of years, without repelling its near-equatorial satellites away from the equator of date: the satellite inclination oscillates but does not show a secular increase. Nor does it show secular decrease, a fact that is relevant to the discussion of the possibility of high-inclination capture of Phobos and Deimos

Strangeness Enhancement in p+Ap+A and S+AS+A Interactions at SPS Energies

The systematics of strangeness enhancement is calculated using the HIJING and VENUS models and compared to recent data on $\,pp\,$, $\,pA\,$ and $\,AA\,$ collisions at CERN/SPS energies ($200A\,\, GeV\,$). The HIJING model is used to perform a {\em linear} extrapolation from $pp$ to $AA$. VENUS is used to estimate the effects of final state cascading and possible non-conventional production mechanisms. This comparison shows that the large enhancement of strangeness observed in $S+Au$ collisions, interpreted previously as possible evidence for quark-gluon plasma formation, has its origins in non-equilibrium dynamics of few nucleon systems. % Strangeness enhancement %is therefore traced back to the change in the production dynamics %from $pp$ to minimum bias $pS$ and central $SS$ collisions. A factor of two enhancement of $\Lambda^{0}$ at mid-rapidity is indicated by recent $pS$ data, where on the average {\em one} projectile nucleon interacts with only {\em two} target nucleons. There appears to be another factor of two enhancement in the light ion reaction $SS$ relative to $pS$, when on the average only two projectile nucleons interact with two target ones.Comment: 29 pages, 8 figures in uuencoded postscript fil

Study of the B^0 Semileptonic Decay Spectrum at the Upsilon(4S) Resonance

We have made a first measurement of the lepton momentum spectrum in a sample of events enriched in neutral B's through a partial reconstruction of B0 --> D*- l+ nu. This spectrum, measured with 2.38 fb**-1 of data collected at the Upsilon(4S) resonance by the CLEO II detector, is compared directly to the inclusive lepton spectrum from all Upsilon(4S) events in the same data set. These two spectra are consistent with having the same shape above 1.5 GeV/c. From the two spectra and two other CLEO measurements, we obtain the B0 and B+ semileptonic branching fractions, b0 and b+, their ratio, and the production ratio f+-/f00 of B+ and B0 pairs at the Upsilon(4S). We report b+/b0=0.950 (+0.117-0.080) +- 0.091, b0 = (10.78 +- 0.60 +- 0.69)%, and b+ = (10.25 +- 0.57 +- 0.65)%. b+/b0 is equivalent to the ratio of charged to neutral B lifetimes, tau+/tau0.Comment: 14 page, postscript file also available at http://w4.lns.cornell.edu/public/CLN