Water regulation of actinomycin-D binding to DNA: the interplay among drug affinity, DNA long-range conformation, and hydration

Actiaomycin-D (actD) binds to natural DNA at two different classes of binding sites, weak and strong. The affinity for these sites is highly dependent on DNA se(sequence and solution conditions, and the interaction appears to be purely entropic driven Although the entropic character of this reaction has been attributed to the release of water molecules upon drug to DNA complex formation, the mechanism by which hydration regulates actD binding and discrimination between different classes of binding sites on natural DNA is still unknown. In this work, we investigate the role of hydration on this reaction using the osmotic stress method. We skew that the decrease of solution water activity, due to the addition of sucrose, glycerol ethylene glycol, and betaine, favors drug binding to the strong binding sites on DNA by increasing both the apparent binding affinity Delta G, and the number of DNA base pairs apparently occupied by the bound drug n(bp/actD). These binding parameters vary linearly with the logarithm of the molar fraction of water in solution log(X-w), which indicates the contribution of water binding to the energetic of the reaction. It is demonstrated that the hydration change measured upon binding increases proportionally to the apparent size of the binding site n(bp/uctD). This indicates that n(bp/actD) measured from the Scatchard plod is a measure of the size of the DNA molecule changing conformation due to ligand binding. We also find that the contribution of DNA deformation, gauged by n(bp/act) to the total free energy of binding Delta G, is given by Delta G = Delta G(local) + n(bp/actD) x delta G(DNA), where Delta G(local), = -8020 +/- 51 cal/mol of actD bound and delta G(DNa) = -24.1 +/- 1.7cal/mol of base pair at 25 degrees C. We interpret Delta G(local), as the energetic contribution due to the direct interactions of actD with the actual tetranucleotide binding site, and it n(bp/actB) X delta G(DNA) as that due to change inconformation, induced by binding, of it n(bp/actD) DNA base pairs flanking the local site. This interpretation is supported by the agreement found between the value of delta G(DNA) and the torsional free energy change measured independently. We conclude suggesting an allosteric model for ligand binding to DNA, such that the increase in binding affinity is achieved by increasing the relaxation of the unfavorable free energy of binding storage at the local site through a larger number of DNA base pairs. The new aspect on this model is that the size of the complex is not fixed but determined by solutions conditions, such as water activity, which modulate the energetic barrier to change helix conformation. These results may suggest that long-range allosteric transitions of duplex DNA are involved in the inhibition of RNA synthesis by actD, and more generally, in the regulation of transcription. (C) 2000 John Wiley & Sons, Inc.

Viscoelastic and Thermal Properties of Collagen-Xanthan Gum and Collagen-Maltodextrin Suspensions During Heating and Cooling

The purpose of this work was to investigate the viscoelastic properties of aqueous suspensions of crude collagen powder extracted from bovine hides and nonsubmitted to the hydrolysis reaction that leads to gelatin. The studied variables included the collagen concentration and the addition of xanthan gum or maltodextrin at varied concentrations during heating/cooling of the mixtures. Differential scanning calorimetry thermograms showed that the addition of polysaccharides decreased the endothermic peak areas observed at the denaturation temperature of collagen. The rheological properties of the pure collagen suspensions were highly dependent on concentration: 4% and 6% collagen suspensions presented a great increase in the storage modulus after heating/cooling, whereas for concentrations of 8% and 10% G' decreased during heating and did not recover its original value after heating/cooling. The frequency sweeps showed that the thermal treatment was responsible by the strengthening of the interactions that formed the polymer network. Addition of 0.1% xanthan gum to collagen suspensions increased the gel strength, especially after heating/cooling of the system, whereas increasing gum concentration to 0.3% resulted in a weaker gel, which could indicate thermodynamic incompatibility between the biopolymers. Mixtures of collagen and maltodextrin resulted in more fluid structures than those obtained with pure collagen at the same collagen concentration and the range of temperatures in which these mixtures behaved as a gel decreased with increasing concentrations of both collagen and maltodextrin, suggesting incompatibilities between the biopolymers.

Daily sperm production and testicular morphometry in goats according to external scrotal conformation

The present study aimed to compare testicular histology and the testicular cell population as well as spermatogenic efficiency in goats with different scrotal conformations. Eighteen goats were divided into 3 groups: Group I - goats without bipartition of the scrotum, Group II - animals with bipartition of the scrotum up to 50% of the testicular length, Group III - goats with scrotal bipartition more than 50% of the testicular length. In goats in Groups I, II and III, the values for the volume density of seminiferous epithelium were 68.9 +/- 0.6%, 71.5 +/- 2.8% and 73.4 +/- 4.7% (P < 0.05), the height of the seminiferous epithelium were 60.2 +/- 4.9 mu m, 61.0 +/- 5.0 mu m and 73.1 +/- 6.6 mu m (P < 0.05), total length of seminiferous tubules found for Groups I, II and III were 2091.9 +/- 27 m, 2172.5 +/- 24.1 m, and 2340.1 14 m (P < 0.05), number of Sertoli and Leydig cells were 1.8 +/- 0.4 x 10(9) and 1.4 +/- 0.1 x 10(9), 2.2 +/- 0.4 and 2.2 +/- 0.7 x 10(9), and 2.5 +/- 0.1 10(9) and 2.3 +/- 0.510(9)(P < 0.05)and daily sperm production observed were 2.1 0.3 x 109, 2.8 0.4 x 109, and 3.1 0.7 x 109 (P < 0.05). In conclusion, goats with greater scrotal bipartition have a greater capacity to produce reproductive cells that is reflected in a greater reproductive potential. (C) 2011 Elsevier B.V. All rights reserved.

Cooling of pacu (Piaractus mesopotamicus) embryos at various stages of development for 6 or 10 hours

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Anomalous metamagnetic-like transition in a FeRh/Fe3Pt interface occurring at T approximate to 120 K in the field-cooled-cooling curves for low magnetic fields

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Conformation and lytic activity of eumenine mastoparan: A new antimicrobial peptide from wasp venom

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Study on the behavior of the Minimum quantity lubricant - MQL technique under different lubricating and cooling conditions when grinding ABNT 4340 steel

The behavior of the minimum quantity lubricant (MQL) technique was analyzed under different lubricating and cooling conditions when grinding ABNT 4340 steel. The comparative analysis of the residual stress values showed that residual compressive stresses were obtained under all the lubrication/cooling conditions and types of abrasive tools employed. The highest residual compressive stress obtained with the aluminum oxide grinding wheel with MQL under the condition of V= 30m/s for air and V= 40ml/h for lubricant was -376MPa against the -160MPa attained with conventional cooling, representing a 135% increase in residual compressive stress. The results show that method and quantity of lubricant and cooling are factors that influence the grinding process.

An evaporative and desiccant cooling system for air conditioning in humid climates

Evaporative cooling operates using water and air as working fluids. It consists in water evaporation, through the passage of an airflow, thus decreasing the air temperature. This system has a great potential to provide thermal comfort in places where air humidity is low, being, however, less efficient where air humidity is high. A way to solve this problem is to use dehumidifiers to pre-conditioning the process air. This paper presents a system that can be used in humid climates coupling desiccant dehumidification equipment to evaporative coolers. The paper shows, initially, the main characteristics of the evaporative cooling and of the adsorption dehumidification systems. Later on the coupled systems, in which occurs a dehumidification by adsorption in a counter flow rotary heat exchanger following the evaporate cooling of the air in evaporative coolers, are analyzed. The thermodynamic equations of state are also presented. Following, this paper analyzes some operation parameters such as: reactivation temperature, R/P relationship (reactivation air flow/ process air flow) and the thermodynamic conditions of the entering air flow. The paper shows the conditions for the best operation point, with regard to thermal comfort conditions and to the energy used in the process. In addition this paper presents an application of the system in different climate characteristics of several tropical and equatorial cities. Copyright © 2005 by ABCM.

A structural conformation study of aquatic humic acid

In the present work, aquatic humic substances (HS) were extracted by use of adsorbent XAD 8 and the acid humic fraction (AH) was separated throught acidification. After being purified by Hyphan resin and dialyze, the aquatic AH was characterized using Fourier-transform infrared spectroscopy and elemental analysis. The influence of the aquatic HA and electrolyte concentrations, pH and aquatic AH-metal complexation time on the conformation was investigated using UV/Vis spectroscopic studies, employing the equation suggested by Doty and Steiner. The results indicated that the acid humic flexible macromolecule assumes a condensed form at acid and alkaline pH. Other factors favoring condensed conformations are longer metal complexation time (ageing) and higher aquatic AH and electrolyte concentrations. Thus considering the strong influence of the investigated parameters in the structural conformation of the humic macromolecule, we conclude that studies using UV/Vis spectroscopy to estimate the concentration, aromaticity, humification degree of the aquatic AH and so on, require rigorous control over the experimental conditions employed to provide a correct interpretation of the analytical results. ©2006 Sociedade Brasileira de Química.

Grinding of hardened steels using optimized cooling

Grinding - the final machining process of a workpiece - requires large amounts of cutting fluids for the lubrication, cooling and removal of chips. These fluids are highly aggressive to the environment. With the technological advances of recent years, the worldwide trend is to produce increasingly sophisticated components with very strict geometric and dimensional tolerances, good surface finish, at low costs, and particularly without damaging the environment. The latter requirement can be achieved by recycling cutting fluids, which is a costly solution, or by drastically reducing the amount of cutting fluids employed in the grinding process. This alternative was investigated here by varying the plunge velocity in the plunge cylindrical grinding of ABNT D6 steel, rationalizing the application of two cutting fluids and using a superabrasive CBN (cubic boron nitride) grinding wheel with vitrified binder to evaluate the output parameters of tangential cutting force, acoustic emission, roughness, roundness, tool wear, residual stress and surface integrity, using scanning electron microscopy (SEM) to examine the test specimens. The performance of the cutting fluid, grinding wheel and plunge velocity were analyzed to identify the best machining conditions which allowed for a reduction of the cutting fluid volume, reducing the machining time without impairing the geometric and dimensional parameters, and the surface finish and integrity of the machined components.

Power losses analysis and cooling system design of three topologies of multilevel inverters

This paper presents the comparison of three topologies of multilevel inverters applied to drive an induction motor of 500 kVA/4.16 kV. The multilevel inverters analyzed are: a neutral point clamped inverter, a symmetrical cascaded multilevel inverter and a hybrid asymmetrical cascaded multilevel inverter. The performance indexes used in the comparison are total harmonic distortion, first order distortion factor, semiconductors power losses distribution and heat-sink volume. The comparison is developed with the purpose of finding the efficiency and the heat-sink volume where the three systems present the same output filter. ©2008 IEEE.

Analysis of agonist and antagonist effects on thyroid hormone receptor conformation by hydrogen/deuterium exchange

Thyroid hormone receptors (TRs) are ligand-gated transcription factors with critical roles in development and metabolism. Although x-ray structures of TR ligand-binding domains (LBDs) with agonists are available, comparable structures without ligand (apo-TR) or with antagonists are not. It remains important to understand apo-LBD conformation and the way that it rearranges with ligands to develop better TR pharmaceuticals. In this study, we conducted hydrogen/deuterium exchange on TR LBDs with or without agonist (T 3) or antagonist (NH3). Both ligands reduce deuterium incorporation into LBD amide hydrogens, implying tighter overall folding of the domain. As predicted, mass spectroscopic analysis of individual proteolytic peptides after hydrogen/ deuterium exchange reveals that ligand increases the degree of solvent protection of regions close to the buried ligand-binding pocket. However, there is also extensive ligand protection of other regions, including the dimer surface at H10-H11, providing evidence for allosteric communication between the ligand-binding pocket and distant interaction surfaces. Surprisingly, Cterminal activation helix H12, which is known to alter position with ligand, remains relatively protected from solvent in all conditions suggesting that it is packed against the LBD irrespective of the presence or type of ligand. T 3, but not NH3, increases accessibility of the upper part of H3-H5 to solvent, and we propose that TR H12 interacts with this region in apo-TR and that this interaction is blocked by T 3 but not NH3.Wepresent data from site-directed mutagenesis experiments and molecular dynamics simulations that lend support to this structural model of apo-TR and its ligand-dependent conformational changes. (Molecular Endocrinology 25: 15-31, 2011). Copyright © 2011 by The Endocrine Society.

Influence of optimized lubrication-cooling and minimum quantity lubrication on the cutting forces, on the geometric quality of the surfaces and on the micro-structural integrity of hardened steel parts

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

Analysis on turning Ti-6Al-4V with different cooling methods

The aim of this paper is to optimize the machining of Ti-6Al-4V alloy, by studying the chip formation, roughness and tool wear for different cooling conditions. The results were compared between cooling methods, minimal quantity of fluid (MQF) and flooding, and also without fluid for the tool H13A. The turning of Ti-6Al-4V has shown good results on roughness (0, 8μm) and tool life, which was 11% lower with MQF than with the flooding method. The tool wear causes variation of the shear angle, which promotes strength hardening of the chip. As a result, the machined surface could be damaged. The use of the cutting fluid helps to save the cutting edge and could reduce the strength hardening. Nevertheless, it could also facilitate the formation of built-up edge. The nucleation of alpha lamellar colonies can occur due to a combination of deformation rates and temperature, mainly when the flooding is used, but it's not conclusive. The lamellar colonies were also found with the MQF in some regions, however, this structure did not show hardness variation compared to equiaxial. For all this reasons, the machining parameters might be carefully chosen.