Crystal structure of a myotoxic Asp49-phospholipase A(2) with low catalytic activity: Insights into Ca2+ -independent catalytic mechanism

A myotoxic Asp49-phospholipase A(2) (Asp49-PLA(2)) with low catalytic activity (BthTX-II from Bothrops jararacussu venom) was crystallized and the molecular-replacement solution has been obtained with a dimer in the asymmetric unit. The quaternary structure of BthTX-II resembles the myotoxic Asp49-PLA2 PrTX-III (piratoxin III from B. pirajai venom) and all non-catalytic and myotoxic dimeric Lys49-PLA(2)s. Despite of this, BthTX-II is different from the highly catalytic and non-myotoxic BthA-I (acidic PLA(2) from B. jararacussu) and other Asp49-PLA(2)s. BthTX-II structure showed a severe distortion of calcium-binding loop leading to displacement of the C-terminal region. Tyr28 side chain, present in this region, is in an opposite position in relation to the same residue in the catalytic activity Asp49-PLA(2)s, making a hydrogen bond with the atom 0 delta 2 of the catalytically active Asp49, which should coordinate the calcium. This high distortion may also be confirmed by the inability of BthTX-II to bind Na+ ions at the Ca2+-binding loop, despite of the crystallization to have occurred in the presence of this ion. In contrast, other Asp49-PLA(2)s which are able to bind Ca2+ ions are also able to bind Na+ ions at this loop. The comparison with other catalytic, non-catalytic and inhibited PLA(2)s indicates that the BthTX-II is not able to bind calcium ions; consequently, we suggest that its low catalytic function is based on an alternative way compared with other PLA(2)s. (c) 2008 Elsevier B.V All rights reserved.

Role of PKC and CaV1.2 in Detrusor Overactivity in a Model of Obesity Associated with Insulin Resistance in Mice

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Diversity and distribution of crustaceans from 13 sheltered sandy beaches along Sao Sebastiao Channel, south-eastern Brazil

The crustacean species composition in the intertidal zones of 13 sheltered unconsolidated marine beaches in south-eastern Brazil is described. Fifty-three crustacean species were collected, adding 46 species to the total reported by previous studies in the same region. Decapods dominated the community, in contrast to exposed sandy beaches where peracarids normally predominate. The species were distributed irregularly among the beaches. Richness varied markedly among sites, and was positively related to a combination of factors such as fine sand grains, high organic matter content, and relatively low silt-clay content. The presence of rock fragments enabled both rocky shore and sandy beach crustaceans to occur oil the same beaches. Richness and abundance of crustaceans showed no clear relationship to sediment grain size and slope, in contrast to the norm for exposed sandy beaches. The dominance of the tanaid Kalliapseudes schubarti in sonic areas may be a result of organic matter pollution in the region. These beaches showed higher species richness than typical sheltered and exposed sandy beaches, indicating that this sheltered, highly heterogeneous seascape is an important area for conservation.

Modal analysis of anisotropic diffused-channel waveguide by a scalar finite element method

A procedure to model optical diffused-channel waveguides is presented in this work. The dielectric waveguides present anisotropic refractive indexes which are calculated from the proton concentration. The proton concentration inside the channel is calculated by the anisotropic 2D-linear diffusion equation and converted to the refractive indexes using mathematical relations obtained from experimental data, the arbitrary refractive index profile is modeled by a. nodal expansion in the base functions. The TE and TM-like propagation properties (effective index) and the electromagnetic fields for well-annealed proton-exchanged (APE) LiNbO3 waveguides are computed by the finite element method.

Insights into metal ion binding in phospholipases A(2): ultra high-resolution crystal structures of an acidic phospholipase A(2) in the Ca2+ free and bound states

The electrophile Ca2+ is an essential multifunctional co-factor in the phospholipase A(2) mediated hydrolysis of phospholipids. Crystal structures of an acidic phospholipase A(2) from the venom of Bothrops jararacussu have been determined both in the Ca2+ free and bound states at 0.97 and 1.60 angstrom resolutions, respectively. In the Ca2+ bound state, the Ca2+ ion is penta-coordinated by a distorted pyramidal cage of oxygen and nitrogen atoms that is significantly different to that observed in structures of other Group I/II phospholipases A(2). In the absence of Ca2+, a water molecule occupies the position of the Ca2+ ion and the side chain of Asp49 and the calcium-binding loop adopts a different conformation. (c) 2005 Elsevier SAS. All rights reserved.

Structure of a Lys49 phospholipase A(2) homologue isolated from the venom of Bothrops nummifer (jumping viper)

Lys49-Phospholipase A(2) (Lys49-PLA(2)) homologues damage membranes by a Ca2+-independent mechanism which does not involve catalytic activity, We have solved the structure of myotoxin-I, a Lys49-PLA(2) homologue isolated from the venom of Bothrops nummifer (jumping viper) at 2.4 Angstrom resolution using molecular replacement techniques. The final model has been refined to a final R-factor of 18.4% (R-free = 23.2%), and shows excellent geometry, the myotoxin-I from Bothrops nummifer is dimeric in the crystalline state as has been observed for other Lys49-PLA(2) homologues. In addition, a continuous electron density in the active site and substrate binding channel could be successfully modeled as a fatty-acid molecule. (C) 1999 Elsevier B.V. Ltd, All rights reserved.

Activation of Ca2+-independent membrane-damaging activity in Lys49-phospholipase A(2) promoted by amphiphilic molecules

Association of class-II phospholipase A(2) (PLA(2)) with aggregated phospholipid substrate results in elevated levels of the Ca2+-dependent hydrolytic activity. The Asp49 residue participates in coordination of the Ca2+ ion cofactor, however, in Lys49-PLA(2) homologues (Lys49-PLA(2)S), substitution of the Asp49 by Lys results in loss of Ca2+ binding and lack of detectable phospholipid hydrolysis. Nevertheless, Lys49-PLA2S cause Ca2+-independent damage of liposome membranes. Bothropstoxin-I is a homodimeric Lys49-PLA(2) from the venom of Bothrops jararacussu, and in fluorescent marker release and dynamic light scattering experiments with DPPC liposomes we demonstrate activation of the Ca2+-independent membrane damaging activity by similar to4 molecules of sodium dodecyl sulphate (SDS) per protein monomer. Activation is accomparlied by significant changes in the intrinsic tryptophan fluorescence emission (ITFE) and near UV circular dichroism (UVCD) spectra of the protein. Subsequent binding of 7-10 SDS molecules results in further alterations in the ITFE and far UVCD spectra. Reduction in the rate of N-bromosuccinimide modification of Trp77 at the dimer interface suggests that initial binding of SDS to this region accompanies the activation of the membrane damaging activity. 1-anilinonaphthalene-8-sulphonic acid binding studies indicate that subsequent SDS binding to the active site is concomitant with the second structural transition. These results provide insights in the structural basis of amphiphile/protein coupling in class-II PLA(2)s. (C) 2004 Published by Elsevier B.V.

Study of the mechanism of action of anoplin, a helical antimicrobial decapeptide with ion channel-like activity, and the role of the amidated C-terminus

Anoplin, an antimicrobial, helical decapeptide from wasp venom, looses its biological activities by mere deamidation of its C-terminus. Secondary structure determination, by circular dichroism spectroscopy in amphipathic environments, and lytic activity in zwitterionic and anionic vesicles showed quite similar results for the amidated and the carboxylated forms of the peptide. The deamidation of the C-terminus introduced a negative charge at an all-positive charged peptide, causing a loss of amphipathicity, as indicated by molecular dynamics simulations in TFE/water mixtures and this subtle modification in a peptide's primary structure disturbed the interaction with bilayers and biological membranes. Although being poorly lytic, the amidated form, but not the carboxylated, presented ion channel-like activity on anionic bilayers with a well-defined conductance step; at approximately the same concentration it showed antimicrobial activity. The pores remain open at trans-negative potentials, preferentially conducting cations, and this situation is equivalent to the interaction of the peptide with bacterial membranes that also maintain a high negative potential inside. Copyright (C) 2007 European Peptide Society and John Wiley & Sons, Ltd.

Measurement of the B-s(0) lifetime in the exclusive decay channel B-s(0)-> J/psi phi

Using the exclusive decay B-s(0)-->J/psi(mu(+)mu(-))phi(K+K-), we report the most precise single measurement of the B-s(0) lifetime. The data sample corresponds to an integrated luminosity of approximately 220 pb(-1) collected with the D0 detector at the Fermilab Tevatron Collider in 2002-2004. We reconstruct 337 signal candidates, from which we extract the B-s(0) lifetime, tau(B-s(0))=1.444(-0.090)(+0.098)(stat)+/-0.020(sys) ps. We also report a measurement for the lifetime of the B-0 meson using the exclusive decay B-0-->J/psi(mu(+)mu(-))K-*0(892)(K(+)pi(-)). We reconstruct 1370 signal candidates, obtaining tau(B-0)=1.473(-0.050)(+0.052)(stat)+/-0.023(sys) ps, and the ratio of lifetimes, tau(B-s(0))/tau(B-0)=0.980(-0.071)(+0.076)(stat)+/-0.003(sys).

Positronium atom scattering by H-2 in a coupled-channel framework

The scattering of ortho-positronium (Ps) by H-2 has been investigated using a three-Ps-state (Ps(1s,2s, 2p)H-2(X (1)Sigma(g)(+))) coupled-channel model and using the Born approximation for higher excitations and ionization of Ps and B (1)Sigma(u)(+) and b (3)Sigma(u)(+) excitations of H-2. We employ a recently proposed time-reversal-symmetric non-local electron-exchange model potential. We present a calculational scheme for solving the body-frame fixed-nuclei coupled-channel scattering equations for Ps-H-2, which simplifies the numerical solution technique considerably. Ps ionization is found to have the leading contribution to target-elastic and all target-inelastic processes. The total cross sections at low and medium energies are in good agreement with experiment.

Collision of positronium with atoms, molecules and ions using a modified coupled-channel framework

Scattering of positronium (Ps) from atoms (H, He, Ne, Ar), molecule (H(2)) and ion (He(+)) have been investigated using a coupled-channel (CC) formalism with a regularised non-local exchange potential. The advantage of using such a regularized exchange potential in the close-coupling formalism and the normalizability aspect of the solution at low energies with a minimum effective coupling are discussed. Results for the elastic and total scattering cross-sections, resonance and binding energies in Ps-H, and pick-off annihilation results in Ps-He are found to be in excellent agreement with measurements and variational predictions. (C) 2000 Elsevier B.V. B.V. All rights reserved.

Measurement of the top quark mass in the lepton plus jets channel using the ideogram method

A measurement of the top quark mass using events with one charged lepton, missing transverse energy, and jets in the final state, collected by the D0 detector from p (p) over bar collisions at root s=1.96 TeV at the Fermilab Tevatron collider, is presented. A constrained fit is used to fully reconstruct the kinematics of the events. For every event a top quark mass likelihood is calculated taking into account all possible jet assignments and the probability that an event is signal or background. Lifetime-based identification of b jets is employed to enhance the separation between t (t) over bar signal and background from other physics processes and to improve the assignment of the observed jets to the quarks in the t (1) over bar hypothesis. We extract a multiplicative jet energy scale (JES) factor in situ, greatly reducing the systematic effect related to the jet energy measurement. In a data sample with an integrated luminosity of 425 pb(-1), we observe 230 candidate events, with an estimated background of 123 events, and measure m(t)=173.7 +/- 4.4(stat+JES)(-2.0)(+2.1)(syst) GeV. This result represents the first application of the ideogram technique to the measurement of the top quark mass in lepton+jets events.

Measurement of the p(p)over-bar -> t(t)over-bar + X production cross section at root s=1.96 TeV in the fully hadronic decay channel

A measurement of the top quark pair production cross section in proton antiproton collisions at an interaction energy of root s=1.96 TeV is presented. This analysis uses 405 +/- 25 pb(-1) of data collected with the D0 detector at the Fermilab Tevatron Collider. Fully hadronic t (t) over bar decays with final states of six or more jets are separated from the multijet background using secondary vertex tagging and a neural network. The t (t) over bar cross section is measured as sigma(t (t) over bar)=4.5(-1.9)(+2.0)(stat)(-1.1)(+1.4)(syst)+/- 0.3(lumi) pb for a top quark mass of m(t)=175 GeV/c(2).

Measurement of the top quark mass in the dilepton channel

We present a measurement of the top quark mass in the dilepton channel based on approximately 370 pb(-1) of data collected by the DO experiment during Run R of the Fermilab Tevatron collider. We employ two different methods to extract the top quark mass. We show that both methods yield consistent results using ensemble tests of events generated with the DO Monte Carlo simulation. We combine the results from the two methods to obtain a top quark mass m(t) = 178.1 +/- 8.2 GeV. The statistical uncertainty is 6.7 GeV and the systematic uncertainty is 4.8 GeV. (c) 2007 Elsevier B.V. All rights reserved.

Search for the standard model Higgs boson in the p(p)over-bar -> ZH -> v(v)over-barb(b)over-bar channel

We report a search for the standard model (SM) Higgs boson based on data collected by the D0 experiment at the Fermilab Tevatron Collider, corresponding to an integrated luminosity of 260 pb(-1). We study events with missing transverse energy and two acoplanar b jets, which provide sensitivity to the ZH production cross section in the nu nu bb channel, and to WH production when the lepton from the W ->center dot nu decay is undetected. The data are consistent with the SM background expectation, and we set 95% C.L. upper limits on sigma(pp -> ZH/WH) x B(H -> bb) from 3.4/8.3 to 2.5/6.3 pb, for Higgs-boson masses between 105 and 135 GeV.