Crystal structure of piratoxin-I: A calcium-independent, myotoxic phospholipase A(2)-homologue from Bothrops pirajai venom

The crystal structure of Piratoxin-I (PrTX-I) a Lys49 homologue isolated from the venom of Bothrops pirajai has been determined and refined at 2.8 Angstrom to a crystallographic residual of 19.7% (R-free = 29.7%). Amino-acid sequence differences between catalytically active phospholipases and PrTX-I in the putative Ca2+-binding loop, specifically the substitutions Tyr28-->Asn, Gly32-->Leu and Asp49-->Lys, result in an altered conformation of this loop, the analysis of the position of the E-amino group of Lys49 in the PrTX-I structure indicates that it fills the site normally occupied by the calcium ion in the catalytically active phospholipases, In contrast to the homologous monomeric Lys49 variant from Agkistrodon piscivorus piscivorus (App), PrTX-I is present as a dimer in the crystalline state, as observed in the structures of myotoxin II from Bothrops asper and Bothropstoxin I from Bothrops jararacussu. The two molecules in the asymmetric unit in the crystal structure of PrTX-I are related by a nearly perfect two-fold symmetry axis, yet the dimeric structure is radically different from the dimeric structure of the phospholipase from Crotalus atrox. In the C. atrox structure the dimer interface occludes the active sites, whereas in the PrTX-I structure they are exposed to solvent, (C) 1998 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.

Cutoff-independent regularization of four-fermion interactions for color superconductivity

We implement a cutoff-independent regularization of four-fermion interactions to calculate the color-superconducting gap parameter in quark matter. The traditional cutoff regularization has difficulties for chemical potentials mu of the order of the cutoff Lambda, predicting in particular a vanishing gap at mu similar to Lambda. The proposed cutoff-independent regularization predicts a finite gap at high densities and indicates a smooth matching with the weak coupling QCD prediction for the gap at asymptotically high densities.

Model-independent measurement of t-channel single top quark production in p(p)over-bar collisions at,root s=1.96 TeV

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Model-independent measurement of the W-boson helicity in top-quark decays at D0

We present the first model-independent measurement of the helicity of W bosons produced in top quark decays, based on a 1 fb(-1) sample of candidate t (t) over bar events in the dilepton and lepton plus jets channels collected by the D0 detector at the Fermilab Tevatron p (p) over bar Collider. We reconstruct the angle theta(*) between the momenta of the down-type fermion and the top quark in the W boson rest frame for each top quark decay. A fit of the resulting cos theta(*) distribution finds that the fraction of longitudinal W bosons f(0)=0.425 +/- 0.166(stat)+/- 0.102(syst) and the fraction of right-handed W bosons f(+)=0.119 +/- 0.090(stat)+/- 0.053(syst), which is consistent at the 30% C.L. with the standard model.

Structure of a calcium-independent phospholipase-like myotoxic protein from Bothrops asper venom

Myotoxin II, a myotoxic calcium-independent phospholipase-like protein isolated from the venom of Bothrops asper, possesses no detectable phospholipase activity. The crystal structure has been determined and refined at 2.8 Angstrom to an R factor of 16.5% (F>3 sigma) with excellent stereochemistry. Amino-acid differences between catalytically active phospholipases and myotoxin LI in the Ca2+-binding region, specifically the substitutions Tyr28-->Asn, Gly32-->Leu and Asp49-->Lys, result in an altered local conformation. The key difference is that the epsilon-amino group of Lys49 fills the site normally occupied by the calcium ion in catalytically active phospholipases. In contrast to the homologous monomeric Lys49 variant from Agkistrodon piscivorus piscivorus, myotoxin II is present as a dimer both in solution and in the crystalline state. The two molecules in the asymmetric unit are related by a nearly perfect twofold axis, yet the dimer is radically different from the dimer formed by the phospholipase from Crotalus atrox. Whereas in C. atrox the dimer interface occludes the active sites, in myotoxin II they are exposed to solvent.

The use of somatosensory information during the acquisition of independent upright stance

This study investigated developmental changes in the use of a contact surface during the acquisition of upright posture. Standing infants were longitudinally examined at four developmental epochs: pulling to stand (PS); standing alone (SA); walking onset (WO); and 1.5 months post-walking (PW). The results revealed that as standing experience increased the force applied to the contact surface by the hand and the body sway decreased. Applied force and body sway were consistently related in the anterior-posterior direction (r approximate to 0.65). Temporally, body sway led applied force (approximate to 45 ms) at the PS, SA, and WO developmental periods. However, at PW, the temporal relationship reversed and applied force led body sway (approximate to 140 ms). These results indicate that initially infants use surface contact for mechanical purposes but later for orientation information that affords prospective control of posture.

ON THE INDEPENDENT-PARTICLE APPROXIMATION OF GAUGE-THEORIES - A SIMPLE EXAMPLE

In this work the independent particle model formulation is studied as a mean-field approximation of gauge theories using the path integral approach in the framework of quantum electrodynamics in 1 + 1 dimensions. It is shown how a mean-field approximation scheme can be applied to fit an effective potential to an independent particle model, building a straightforward relation between the model and the associated gauge field theory. An example is made considering the problem of massive Dirac fermions on a line, the so called massive Schwinger model. An interesting result is found, indicating a behaviour of screening of the charges in the relativistic limit of strong coupling. A forthcoming application of the method developed to confining potentials in independent quark models for QCD is in view and is briefly discussed.

Adaptive control charts: A Markovian approach for processes subject to independent disturbances

Recent studies have shown that adaptive X control charts are quicker than traditional X charts in detecting small to moderate shifts in a process. In this article, we propose a joint statistical design of adaptive X and R charts having all design parameters varying adaptively. The process is subjected to two independent assignable causes. One cause changes the process mean and the other changes the process variance. However, the occurrence of one kind of assignable cause does not preclude the occurrence of the other. It is assumed that the quality characteristic is normally distributed and the time that the process remains in control has exponential distribution. Performance measures of these adaptive control charts are obtained through a Markov chain approach. (c) 2005 Elsevier B.V. All rights reserved.

Maximal lactate steady state concentration independent of pedal cadence in active individuals

The maximal lactate steady state (MLSS) is defined as the highest blood lactate concentration that can be maintained over time without a continual blood lactate accumulation. The objective of the present study was to analyze the effects of pedal cadence (50 vs. 100 rev min(-1)) on MLSS and the exercise workload at MLSS (MLSSworkload) during cycling. Nine recreationally active males (20.9 +/- 2.9 years, 73.9 +/- 6.5 kg, 1.79 +/- 0.09 m) performed an incremental maximal load test (50 and 100 rev min(-1)) to determine anaerobic threshold (AT) and peak workload (PW), and between two and four constant submaximal load tests (50 and 100 rev min(-1)) on a mechanically braked cycle ergometer to determine MLSSworkload and MLSS. MLSSworkload was defined as the highest workload at which blood lactate concentration did not increase by more than 1 mM between minutes 10 and 30 of the constant workload. The maximal lactate steady state intensity (MLSSintensity) was defined as the ratio between MLSSworkload and PW. MLSSworkload (186.1 +/- 21.2 W vs. 148.2 +/- 15.5 W) and MLSSintensity (70.5 +/- 5.7% vs. 61.4 +/- 5.1%) were significantly higher during cycling at 50 rev min(-1) than at 100 rev min(-1), respectively. However, there was no significant difference in MLSS between 50 rev min(-1) (4.8 +/- 1.6 mM) and 100 rev min(-1) (4.7 +/- 0.8 mM). We conclude that MLSSworkload and MLSSintensity are dependent on pedal cadence (50 vs. 100 rev min(-1)) in recreationally active individuals. However, this study showed that MLSS is not influenced by the different pedal cadences analyzed.

Calcium-Independent Membrane Damage by Venom Phospholipases A(2)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

K-ATP(+) Channels-Independent Analgesic Action of Crotalus durissus cumanensis venom

The effect was investigated of the K+ channel blocker, glibenclamide, on the ability of Crotalus durissus cumanensis venom (CDCM) to promote peripheral antinociception. This was measured by formalin-induced nociception in male Swiss mice. CDCM (200 and 300 mu g/kg) produced an antinociceptive effect during phase 2 in the formalin test. The effect of CDCM (200 mu g/kg) was unaffected by the ATP-sensitive K+ channel blocker glibenclamide (2 mg/kg). These results suggest that CDCM is effective against acute pain. However, the ATP-sensitive K+ channels pathway is not contributable to the antinoeiceptive mechanism of CDCM.

Heme-Dependent and Independent Soluble Guanylate Cyclase Activators and Vasodilation

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