Expression, purification, and circular dichroism analysis of human CDK9

The human cyclin-dependent kinase 9 (CDK9) protein was expressed in E coli BL21 using the pET23a vector at 30 degrees C. Several milligrams of protein were purified from soluble fraction using ionic exchange and ATP-affinity chromatography. The structural quality of recombinant CDK9 and the estimation of its secondary structure were obtained by circular dichroism. Structural models of CDK9 presented 26% of helices in agreement with the spectra by circular dichroism analysis. This is the first report on human CDK9 expression in Escherichia coli and structure analysis and provides the first step for the development of CDK9 inhibitors. (c) 2006 Elsevier B.V. All rights reserved.

Superconducting boundary conditions for mesoscopic circular samples

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

Temperature-dependent vortex matter in a superconducting mesoscopic circular sector

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

Vortices in a mesoscopic superconducting circular sector

In the present paper we develop an algorithm to solve the time dependent Ginzburg-Landau equations, by using the link variables technique, for circular geometries. In addition, we evaluate the Helmholtz and Gibbs free energy, the magnetization, and the number of vortices. This algorithm is applied to a circular sector. We evaluate the superconduting-normal magnetic field transition, the magnetization, and the superconducting density. Our results point out that, as we reduce the superconducting area, the nucleation field increases. Nevertheless, as the angular width of the circular sector goes to small values the asymptotic behavior is independent of the sample area. We also show that the value of the first nucleation field is approximately the same independently of the form of the circular sector. Furthermore, we study the nucleation of giant and multivortex states for the different shapes of the present geometry.

Low-altitude, near-polar and near-circular orbits around Europa

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

Treatment of segmental tibial defects using acute bone shortening followed by gradual lengthening with circular external fixator

The aim of this study was to clinically and radiographically evaluate acute bone shortening followed by gradual lengthening in the treatment of large segmental tibia defects induced in seven clinically normal dogs. A circular external fixator was assembled with one proximal 5/8-circle ring, one middle ring and one distal ring connected with three rods. Thirty per cent of the tibia and fibula were removed in the middle and distal parts of the diaphyses, between the middle and distal rings. Acute bone shortening with compression of proximal and distal segments was performed. A subperiosteal osteotomy was performed between the half-ring and middle ring. Bone distraction started 7 days after surgery; after lengthening, the apparatus was left in place for 14 weeks for consolidation of regenerated bone. The frame was removed at the end of this period, and the dogs observed for four more weeks. Functional results were considered excellent in two, good in three and fair in the other two dogs. Bone regeneration within the distraction gap was obtained 14 weeks after neutral fixation period. We concluded that acute bone shortening followed by gradual lengthening by Ilizarov method can be used to treat extensive tibial defects in dogs, although it presents limb temporary abnormal limb shape and unequal length as early disadvantages.

A stability transition at mildly acidic pH in the alpha-hemolysin (alpha-toxin) from Staphylococcus aureus

The effects of mildly acidic conditions on the free energy of unfolding (Delta G(u)(buff)) of the pore-forming alpha-hemolysin (alpha HL) from Staphylococcus aureus were assessed between pH 5.0 and 7.5 by measuring intrinsic tryptophan fluorescence, circular dichroism and elution time in size exclusion chromatography during urea denaturation, Decreasing the pH from 7.0 to 5.0 reduced the calculated Delta G(u)(buff) from 8.9 to 4.2 kcal moI(-1), which correlates with an increased rate of pore formation previously observed over the same pH range, It is proposed that the lowered surface pH of biological membranes reduces the stability of alpha HL thereby modulating the rate of pore formation. (C) 1999 Federation of European Biochemical Societies.

Selectivity in the mechanism of action of antimicrobial mastoparan peptide Polybia-MP1

Many potent antimicrobial peptides also present hemolytic activity, an undesired collateral effect for the therapeutic application. Unlike other mastoparan peptides, Polybia-MP1 (IDWKKLLDAAKQIL), obtained from the venom of the social wasp Polybia paulista, is highly selective of bacterial cells. The study of its mechanism of action demonstrated that it permeates vesicles at a greater rate of leakage on the anionic over the zwitterionic, impaired by the presence of cholesterol or cardiolipin; its lytic activity is characterized by a threshold peptide to lipid molar ratio that depends on the phospholipid composition of the vesicles. At these particular threshold concentrations, the apparent average pore number is distinctive between anionic and zwitterionic vesicles, suggesting that pores are similarly formed depending on the ionic character of the bilayer. To prospect the molecular reasons for the strengthened selectivity in Polybia-MP1 and its absence in Mastoparan-X, MD simulations were carried out. Both peptides presented amphipathic alpha-helical structures, as previously observed in Circular Dichroism spectra, with important differences in the extension and stability of the helix; their backbone solvation analysis also indicate a different profile, suggesting that the selectivity of Polybia-MP1 is a consequence of the distribution of the charged and polar residues along the peptide helix, and on how the solvent molecules orient themselves according to these electrostatic interactions. We suggest that the lack of hemolytic activity of Polybia-MP1 is due to the presence and position of Asp residues that enable the equilibrium of electrostatic interactions and favor the preference for the more hydrophilic environment.

Effects of Resinous Monomers Used in Restorative Dental Modeling on the Cohesive Strength of Composite Resin

Purpose: The purpose of this study was to evaluate the cohesive strength of the composite using different resinous monomers to lubricate instruments used in the Restorative Dental Modeling Insertion Technique (RDMIT).Materials and Methods: The composite specimens were made by using a prefabricated Teflon device. Different resinous monomers were used at the interface to lubricate the instruments, for a total of 72 specimens divided into 6 groups: 1. control group, no resinous monomer was used; 2. Composite Wetting Resin; 3. C & B Liquid; 4. Scotchbond Multi-Purpose Adhesive; 4. Adper Single Bond Adhesive; 6. Prime & Bond NT. Specimens were submitted to the circular area tensile test to evaluate the cohesive strength at the composite interfaces. Data were analyzed using ANOVA and Tukey's test (alpha = 0.05).Results: ANOVA showed a value of p < 0.0001, which indicated that there were significant differences among the groups. The means (SD) for the different groups were: Adper Single Bond Adhesive: 26 (12) a; control group: 28 (3) ab; Prime & Bond NT: 32 (12) ab; Composite Wetting Resin: 36 (9) abc; C&B Liquid: 38 (7) bc; Scotchbond Multi-Purpose Adhesive: 46 (10) c. Groups denoted with the same letters were not significantly different. Only Scotchbond Multi-Purpose Adhesive, used for direct restorations, had a statistically significantly higher bond strength than the control group, Adper Single Bond Adhesive, and Prime & Bond NT. Adper Single Bond with Adhesive showed a statistically significantly lower mean value than C & B Liquid.Conclusion: The results of this study indicate that the resinous monomers used for lubricating the instruments in the RDMIT did not alter the mechanical properties of the composite, and therefore did not reduce the cohesive bond strength at the composite interfaces.

Semiclassical approach to the decay of protons in circular motion under the influence of gravitational fields

We investigate the possible decay of protons in geodesic circular motion around neutral compact objects. Weak and strong decay rates and the associated emitted powers are calculated using a semiclassical approach. Our results are discussed with respect to distinct ones in the literature, which consider the decay of accelerated protons in electromagnetic fields. A number of consistency checks are presented along the paper.

Mechanism of Action and Relationship Between Structure and Biological Activity of Ctx-Ha: A New Ceratotoxin-like Peptide from Hypsiboas albopunctatus

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Further monoterpene chromane esters from Peperomia obtusifolia: VCD determination of the absolute configuration of a new diastereomeric mixture

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

Structure and absolute configuration of a secolignan from Peperomia blanda

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