Quality characteristics and fertilizing ability of ram sperm subpopulations separated by partition in an aqueous two-phase system

Centrifugal countercurrent distribution (CCCD) in an aqueous two-phase system (TPS) is a resolute technique revealing sperm heterogeneity and for the estimation of the fertilizing potential of a given semen sample. However, separated sperm subpopulations have never been tested for their fertilizing ability yet. Here, we have compared sperm quality parameters and the fertilizing ability of sperm subpopulations separated by the CCCD process from ram semen samples maintained at 20 degrees C or cooled down to 5 degrees C. Total and progressive sperm motility was evaluated by computer-assisted analysis using a CASA system and membrane integrity was evaluated by flow cytometry by staining with CFDA/Pl. The capacitation state, staining with chlortetracycline, and apoptosis-related markers, such as phosphatidylserine (PS) translocation detected with Annexin V. and DNA damage detected by the TUNEL assay, were determined by fluorescence microscopy. Additionally, the fertilizing ability of the fractionated subpopulations was comparative assessed by zona binding assay (ZBA). CCCD analysis revealed that the number of spermatozoa displaying membrane and DNA alterations was higher in samples chilled at 5 degrees C than at 20 degrees C. which can be reflected in the displacement to the left of the CCCD profiles. The spermatozoa located in the central and right chambers (more hydrophobic) presented higher values (P<0.01) of membrane integrity, lower PS translocation (P<0.05) and DNA damage (P<0.001) than those in the left part of the profile, where apoptotic markers were significantly increased and the proportion of viable non-capacitated sperm was reduced. We have developed a new protocol to recover spermatozoa from the CCCD fractions and we proved that these differences were related with the fertilizing ability determined by ZBA, because we found that the number of spermatozoa attached per oocyte was significantly higher for spermatozoa recovered from the central and right chambers, in both types of samples. This is the first time, to our knowledge that sperm recovered from a two-phase partition procedure are used for fertilization assays. These results open up new possibilities for using specific subpopulations of sperm for artificial insemination or in vitro fertilization, not only regarding better sperm quality but also certain characteristics such as subpopulations enriched in spermatozoa bearing X or Y chromosome that we have already isolated or any other feature. (C) 2011 Elsevier B.V. All rights reserved.

Enantioselective analysis of unbound tramadol, O-desmethyltramadol and N-desmethyltramadol in plasma by ultrafiltration and LC-MS/MS: Application to clinical pharmacokinetics

This study describes the enantioselective analysis of unbound and total concentrations of tramadol and its main metabolites O-desmethyltramadol (M1) and N-desmethyltramadol (M2) in human plasma. Sample preparation was preceded by an ultrafiltration step to separate the unbound drug. Both the ultrafiltrate and plasma samples were submitted to liquid/liquid extraction with methyl t-butyl ether. Separation was performed on a Chiralpak (R) AD column and tandem mass spectrometry consisting of an electrospray ionization source, positive ion mode and multiple reaction monitoring was used as the detection system. Linearity was observed in the following ranges: 0.2-600 and 0.5-250 ng/mL for analysis of total and unbound concentrations of the tramadol enantiomers, respectively, and 0.1-300 and 0.25-125 ng/mL for total and unbound concentrations of the M1 and M2 enantiomers, respectively. The lower limits of quantitation were 0.2 and 0.5 ng/mL for analysis of total and unbound concentration of each tramadol enantiomer, respectively, and 0.1 and 0.25 ng/mL for total and unbound concentrations of M1 and M2 enantiomers, respectively. Intra- and interassay reproducibility and inaccuracy did not exceed 15%. Clinical application of the method to patients with neuropathic pain showed plasma accumulation of (+)-tramadol and (+)-M2 after a single oral dose of racemic tramadol. Fractions unbound of tramadol, M1 or M2 were not enantioselective in the patients investigated. (C) 2011 Elsevier B.V. All rights reserved.

Strategies in identifying individuals in a segregant population of common bean and implications of genotype x environment interaction in the success of selection

The objective of this study was to compare the BLUP selection method with different selection strategies in F-2:4 and assess the efficiency of this method on the early choice of the best common bean (Phaseolus vulgaris) lines. Fifty-one F-2:4 progenies were produced from a cross between the CVIII8511 x RP-26 lines. A randomized block design was used with 20 replications and one-plant field plots. Character data on plant architecture and grain yield were obtained and then the sum of the standardized variables was estimated for simultaneous selection of both traits. Analysis was carried out by mixed models (BLUP) and the least squares method to compare different selection strategies, like mass selection, stratified mass selection and between and within progeny selection. The progenies selected by BLUP were assessed in advanced generations, always selecting the greatest and smallest sum of the standardized variables. Analyses by the least squares method and BLUP procedure ranked the progenies in the same way. The coincidence of the individuals identified by BLUP and between and within progeny selection was high and of the greatest magnitude when BLUP was compared with mass selection. Although BLUP is the best estimator of genotypic value, its efficiency in the response to long term selection is not different from any of the other methods, because it is also unable to predict the future effect of the progenies x environments interaction. It was inferred that selection success will always depend on the most accurate possible progeny assessment and using alternatives to reduce the progenies x environments interaction effect.

Use of Hg-Electroplated-Pt Ultramicroelectrode for Determining Elemental Sulphur in Naphtha Samples

This paper describes the applicability of a Hg-electroplated-Pt ultramicroelectrode in the quantification of elemental sulphur in naphtha samples by square-wave voltammetry. A reproducible deposition methodology was studied and is reported in this paper. This methodology is innovative and relies on the quality of the mercury stock solution to obtain reproducible surfaces required for the analytical methodology. All analyses were performed using a Hg-electroplated-Pt ultramicroelectrode (Hg-Pt UME) due to the low sensibility of such devices to ohmic drops in resistive solutions. The responses of the peak areas in voltammetric experiments were linear in all of the range studied. The method developed here is accurate and reproducible, with a detection limit of 0.010 mg L-1 and a good recovery range for both standard solutions of elemental sulphur (85 to 99%) and real naphtha sample (79%). These results attest to the potential for the application of this electroanalytical methodology in determining elemental sulphur in naphtha samples containing mercaptans and disulphides.

Photoluminescence in silicon/silicon oxide films produced by the Pulsed Electron Beam Ablation technique

In this work we report studies of the photoluminescence emission in samples based on Si/SiOx films deposited by the Pulsed Electron Beam Ablation (PEBA) technique. The samples were prepared at room temperature using targets with different Si/SiO2 concentrations. The samples were characterized using X-ray Absorption Edge Spectroscopy (XANES) at the Si-K edge, Raman spectroscopy, Photoluminescence (PL) and X-ray Photoelectron Spectroscopy (XPS). The concentration of a-Si and nc-Si in the film was dependent on the silicon concentration in the target. It was also observed that the PL is strongly dependent on the structural amorphous/crystalline arrangement. Crown Copyright (C) 2011 Published by Elsevier B.V. All rights reserved.

Costs of most frequent nursing activities in highly dependent hospitalized patients

This quantitative study aimed to identify the costs of the most frequent nursing activities in highly dependent hospitalized patients at a medical clinic. The non-probabilistic convenience sample corresponded to 607 observations regarding oral feeding activities (OF), blood pressure verification (BP)/heart rate (HR), body temperature checking (BTC), performance of intimate hygiene and management of feeding probe. The costs identified corresponded to R$2.40 (SD+/-2.64) for OF feeding; R$1.26 (SD+/-0.48) to verify the BP/HR; R$1.17 (SD+/-0.46) for BTC; R$15.59 (SD+/-8.62) to perform intimate hygiene and R$5.95 (SD+/-2.13) for management of feeding probe. This study will facilitate cost management, with a view to avoiding waste related to unnecessary resource consumption and establish a correlation between costs and care delivery results. Supported by Pro-Reitoria de Pesquisa, Universidade de Sao Paulo, Brazil.

Apoptotic Cells Contribute to Melanoma Progression and This Effect is Partially Mediated by the Platelet-Activating Factor Receptor

There is evidence that the platelet-activating factor receptor (PAFR) is involved in the clearance of apoptotic cells by macrophages, and that this is associated with anti-inflammatory phenotype. Our group has previously shown that coinjection of a large number of apoptotic cells can promote tumor growth from a subtumorigenic dose of melanoma cells. Here, we studied the involvement of the PAFR in the tumor growth promoting effect of apoptotic cells. A sub-tumorigenic dose of melanoma cells (Tm1) was coinjected with apoptotic Tm1 cells, subcutaneously in the flank of C57Bl/6 mice, and the volume was monitored for 30 days. Animals received the PAFR antagonists, WEB2170 or PCA4248 (5 mg/kg body weight) or vehicle, by peritumoral daily injection for 5 days. Results showed that PAFR antagonists significantly inhibited the tumor growth induced by the coinjection of a subtumorigenic dose of melanoma cells together with apoptotic cells. This was accompanied by inhibition of early neutrophil and macrophage infiltration. Addition of (platelet-activating factor) to this system has no significant effect. PAFR antagonists did not affect the promoting effect of carrageenan. We suggest that the recognition of apoptotic cells by phagocytes leads to activation of PAFR pathways, resulting in a microenvironment response favorable to melanoma growth.

Spin-polarized transport in II-VI diluted magnetic semiconductors superlattices

We studied the spin-polarized charge densities in II-VI-based diluted magnetic superlattices formed of p-doped ZnTe:Mg/ZnTe:TM/ZnTe:Mg non-magnetic/magnetic/non-magnetic layers, with TM standing for transition metal. The calculations were performed within a self-consistent k.p method, in which are also taken into account the exchange correlation effects in the local density approximation. Our results show a limit for the width of the non-magnetic layer for which the difference between the opposite spin charge densities is maximized, indicating the best conditions to obtain full polarization by varying the TM content. We also discuss these effects in the calculated photoluminescence spectra. Our findings point to the possibility of engineering the spin-polarized charge distribution by varying the widths of the magnetic and non-magnetic layers and/or varying the TM concentration in the magnetic layers, thus providing a guide for future experiments. (c) 2012 Elsevier B.V. All rights reserved.

Materials prepared from biopolyethylene and curaua fibers: Composites from biomass

Composites of high-density biopolyethylene (HDBPE) obtained from ethylene derived from sugarcane ethanol and curaua fibers were formed by first mixing in an internal mixer followed by thermopressing. Additionally, hydroxyl-terminated polybutadiene (LHPB), which is usually used as an impact modifier, was mainly used in this study as a compatibilizer agent. The fibers, HDBPE and LHPB were also compounded using an inter-meshing twin-screw extruder and, subsequently, injection molded. The presence of the curaua fibers enhanced some of the properties of the HDBPE, such as its flexural strength and storage modulus. SEM images showed that the addition of LHPB improved the adhesion of the fiber/matrix at the interface, which increased the impact strength of the composite. The higher shear experienced during processing probably led to a more homogeneous distribution of fibers, making the composite that was prepared through extruder/injection molding more resistant to impact than the composite processed by the internal mixer/thermopressing. (c) 2012 Elsevier Ltd. All rights reserved.

Intervention Based Exclusively on Stage- Matched Printed EducationalMaterials Regarding Healthy Eating Does Not Result in Changes to Adolescents' Dietary Behavior

Purpose. To assess the impact of a six-month stage-based intervention on fruit and vegetable intake, regarding perceived benefits and barriers, and self-efficacy among adolescents. Design. Randomized treatment-control, pre-post design. Subjects/ Setting. Schools were randomized between control and experimental groups. 860 adolescents from ten public schools in Bras ' ilia, Federal District, Brazil were evaluated at baseline; 771 (81%) completed the study. Intervention. Experimental group received monthly magazines and newsletters aimed at promotion of healthy eating. Measures. Self-reported fruit and vegetable intake, stages of change, self-efficacy and decisional balance scores were evaluated at baseline and post-intervention in both groups. Analysis. The effectiveness of the intervention was evaluated using the analysis of covariance model (ANCOVA) and repeated measurement analysis by means of weighted least squares. Comparison between the proportions of adolescents who advanced through the stages during the intervention was performed using the Mantel-Haenszel chi-square test. Results. After adjusting for sex and age, study variables showed no modifications through the proposed intervention. There was no statistical difference in participant mobility in the intervention and control groups between the stages of change, throughout the study. Conclusion. A nutritional intervention based exclusively on distribution of stage-matched printed educational materials was insufficient to change adolescents' dietary behavior.

Clinical Diagnosis of Pulp Inflammation Based on Pulp Oxygenation Rates Measured by Pulse Oximetry

Introduction: The objective of this study was to investigate correlations between pulp oxygenation rates (%SpO(2)) and clinical diagnoses of reversible pulpitis (RP), irreversible pulpitis (IP), or pulp necrosis (PN). Methods: Sixty patients who presented with a tooth with endodontic pathology were grouped according to a clinical diagnosis of either RP (n = 20), IP (n = 20), or PN (n = 20). The clinical diagnosis was based on the patient's dental history, periapical radiographs, clinical inspection, and percussion and thermal sensitivity testing. Pulse oximetry (PO) was used to determine pulp oxygenation rates. For every patient, one additional endodontically treated tooth (negative control [NC], n = 60) and one additional healthy tooth with healthy pulp status (positive control [PC], n = 60) were evaluated. Analysis of variance, the Tukey HSD test, and the Student's t test were used for statistical analysis. Results: The mean % SpO(2) levels were as follows: RP: 87.4% (standard deviation [SD] +/- 2.46), IP: 83.1% (SD +/- 2.29), PN: 74.6% (SD +/- 1.96), PC: 92.2% (SD +/- 1.84), and NC: 0% (SD +/- 0.0). There were statistically significant differences between RP, IP, and PM compared with NC and PC and between RP, IP, and PN (all P <= .01). Conclusions: The evaluation of pulp oxygenation rates by PO may be a useful tool to determine the different inflammatory stages of the pulp to aid in endodontic diagnosis. (JEndod 2012;38:880-883)

Spinal cord injury as a trigger to develop periodic leg movements during sleep: an evolutionary perspective

The primary trigger to periodic limb movement (PLM) during sleep is still unknown. Its association with the restless legs syndrome (RLS) is established in humans and was reported in spinal cord injury (SCI) patients classified by the American Spinal Injury Association (ASIA) as A. Its pathogenesis has not been completely unraveled, though recent advances might enhance our knowledge about those malfunctions. PLM association with central pattern generator (CPG) is one of the possible pathologic mechanisms involved. This article reviewed the advances in PLM and RLS genetics, the evolution of CPG functioning, and the neurotransmitters involved in CPG, PLM and RLS. We have proposed that SCI might be a trigger to develop PLM.

Seasonal changes in the pigment composition of natural population of Gracilaria domingensis (Gracilariales, Rhodophyta)

The concentrations of phycobiliproteins (phycoerythrin and phycocyanin), chlorophyll-a and total soluble proteins were determined monthly in three strains (red, green and brown) of Gracilaria domingensis (Kützing) Sonder ex Dickie, collected from natural populations on the coast of Rio Grande do Norte, Brazil. In all the strains, pigment and protein concentrations were higher in the months of less sunlight and greater nitrogen availability and decreased gradually with increased sunlight and decreased nutrient concentration. The red strain showed higher concentrations of phycoerythrin and total soluble proteins. The difference in the concentration of biochemical components over the course of the year indicates species acclimation to different environmental conditions.

Mapeamento do índice de vegetação da diferença normalizada em lavoura de algodão

Os dados de sensoriamento remoto em campo podem fornecer informações detalhadas sobre a variabilidade de parâmetros biofísicos ligados à produtividade em grandes áreas e apresentam potencial para o monitoramento destes parâmetros, ao longo de todo o ciclo de desenvolvimento da cultura. Este trabalho objetivou mapear a variabilidade espacial do índice de vegetação da diferença normalizada (NDVI) e seus componentes, em duas lavouras comerciais de algodão (Gossipium hirsutum L.), utilizando sensor óptico ativo, em nível terrestre. Os dados foram coletados utilizando-se sensor instalado em um pulverizador autopropelido agrícola. Um receptor GPS foi acoplado ao sensor, para a obtenção das coordenadas dos pontos de amostragem. As leituras foram realizadas em faixas espaçadas em 21,0 m, aproveitando-se as passadas do veículo no momento da pulverização de agroquímicos, e os dados submetidos à análise estatística clássica e geoestatística. Mapas de distribuição espacial das variáveis foram elaborados pela interpolação por krigagem. Observou-se maior variabilidade espacial do NDVI e da reflectância espectral da vegetação na região do infravermelho próximo (IVP) (880 nm) e do visível (590 nm) na lavoura com maior estresse fisiológico, devido ao ataque do percevejo castanho [Scaptocoris castanea (Hem.: Cydnidae)], em relação à lavoura sadia.

Multi-objective evolutionary algorithm for single and multiple fault service restoration in large-scale distribution systems

Network reconfiguration for service restoration (SR) in distribution systems is a complex optimization problem. For large-scale distribution systems, it is computationally hard to find adequate SR plans in real time since the problem is combinatorial and non-linear, involving several constraints and objectives. Two Multi-Objective Evolutionary Algorithms that use Node-Depth Encoding (NDE) have proved able to efficiently generate adequate SR plans for large distribution systems: (i) one of them is the hybridization of the Non-Dominated Sorting Genetic Algorithm-II (NSGA-II) with NDE, named NSGA-N; (ii) the other is a Multi-Objective Evolutionary Algorithm based on subpopulation tables that uses NDE, named MEAN. Further challenges are faced now, i.e. the design of SR plans for larger systems as good as those for relatively smaller ones and for multiple faults as good as those for one fault (single fault). In order to tackle both challenges, this paper proposes a method that results from the combination of NSGA-N, MEAN and a new heuristic. Such a heuristic focuses on the application of NDE operators to alarming network zones according to technical constraints. The method generates similar quality SR plans in distribution systems of significantly different sizes (from 3860 to 30,880 buses). Moreover, the number of switching operations required to implement the SR plans generated by the proposed method increases in a moderate way with the number of faults.