In vitro development of cloned bovine embryos produced by handmade cloning using somatic cells from distinct levels of cell culture confluence

The relationship between the level of cell confluence near the plateau phase of growth and blastocyst yield following somatic cell cloning is not well understood. We examined the effect of distinct cell culture confluence levels on in vitro development of cloned bovine embryos. In vitro-matured bovine oocytes were manually bisected and selected by DNA staining. One or two enucleated hemi-cytoplasts were paired and fused with an adult skin somatic cell. Cultured skin cells from an adult Nellore cow harvested at three distinct culture confluence levels (70-80, 80-90, and > 95%) were used for construction of embryos and hemi-embryos. After activation, structures were cultured in vitro as one embryo (1 x 100%) or as aggregates of two hemi-embryos (2 x 50%) per microwell. Fusion, cleavage and blastocyst rates were compared using the chi(2) test. The fusion rate for hemi-embryos (51.4%) was lower than for embryos (67.6%), with no influence of degree of cell confluence. However, blastocyst rates improved linearly (7.0, 17.5, and 29.4%) with increases in cell confluence. We conclude that degree of cell culture confluence significantly influences subsequent embryo development; use of a cell population in high confluence (> 90%) for nuclear transfer significantly improved blastocyst yield after cloning.

Serum-Starved Apoptotic Fibroblasts Reduce Blastocyst Production but Enable Development to Term after SCNT in Cattle

Cell cycle synchronization by serum starvation (SS) induces apoptosis in somatic cells. This side effect of SS is hypothesized to negatively affect the outcome of somatic cell nuclear transfer (SCNT). We determined whether apoptotic fibroblasts affect SCNT yields. Serum-starved, adult, bovine fibroblasts were stained with annexin V-FITC/propidium iodide to allow apoptosis detection by flow cytometry. Positive and negative cells sorted by fluorescence activated cell sorting (FACS) and an unsorted control group were used as nuclear donors for SCNT. Reconstructed embryos were cultured in vitro and transferred to synchronized recipients. Apoptosis had no effect on fusion and cleavage rates; however, it resulted in reductions in blastocyst production and quality measured by apoptotic index. However, reconstructed embryos with apoptotic cells resulted in pregnancy rates similar to that of the control on day 30, and generated one live female calf. In conclusion, we showed that apoptotic cells present in serum-starved cultures negatively affect embryo production after SCNT without compromising full-term development. Further studies will evaluate the ability of the oocyte to reprogram cells in specific phases of apoptosis.

Modeling Two-Oscillator Circadian Systems Entrained by Two Environmental Cycles

Several experimental studies have altered the phase relationship between photic and non-photic environmental, 24 h cycles (zeitgebers) in order to assess their role in the synchronization of circadian rhythms. To assist in the interpretation of the complex activity patterns that emerge from these ""conflicting zeitgeber'' protocols, we present computer simulations of coupled circadian oscillators forced by two independent zeitgebers. This circadian system configuration was first employed by Pittendrigh and Bruce (1959), to model their studies of the light and temperature entrainment of the eclosion oscillator in Drosophila. Whereas most of the recent experiments have restricted conflicting zeitgeber experiments to two experimental conditions, by comparing circadian oscillator phases under two distinct phase relationships between zeitgebers (usually 0 and 12 h), Pittendrigh and Bruce compared eclosion phase under 12 distinct phase relationships, spanning the 24 h interval. Our simulations using non-linear differential equations replicated complex non-linear phenomena, such as ""phase jumps'' and sudden switches in zeitgeber preferences, which had previously been difficult to interpret. Our simulations reveal that these phenomena generally arise when inter-oscillator coupling is high in relation to the zeitgeber strength. Manipulations in the structural symmetry of the model indicated that these results can be expected to apply to a wide range of system configurations. Finally, our studies recommend the use of the complete protocol employed by Pittendrigh and Bruce, because different system configurations can generate similar results when a ""conflicting zeitgeber experiment'' incorporates only two phase relationships between zeitgebers.

Phase-disorder-induced double resonance of neuronal activity

It is well known that resonance can be induced by external noise or diversity. Here we show that resonance can be induced even by a phase disorder in coupled excitable neurons with subthreshold activity. In contrast to the case of identical phase, we find that phase disorder plays an active role in enhancing neuronal activity. We also uncover that the presence of phase disorder can induce a double resonance phenomenon: phase disorder and coupling strength both can enhance neuronal firing activity. A physical theory is formulated to help understand the mechanism behind this double resonance phenomenon.

Bicoherence in electrostatic turbulence driven by high magnetohydrodynamic activity in Tokamak Chauffage Alfven Bresilien

During some discharges in Tokamak Chauffage Alfven Bresilien [R. M. O. Galvao et al., Plasma Phys. Controlled Fusion 43, 1181 (2001)] high magnetohydrodynamic activity may appear with a peaked frequency spectrum. Whenever this peak occurs, the ambient broadband electrostatic turbulence is remarkably modified, synchronizing into the dominant magnetic fluctuation frequency and presenting high bicoherence in the whole plasma edge with a maximum bicoherence inside the plasma. A phenomenological model is introduced to investigate this driven turbulence bicoherence, consisting of nonlinearly coupled phase-randomized drift modes with time-periodic external driving at the dominant magnetic fluctuation frequency. The bicoherence spectrum of this model can mimic features of the experimental results. (C) 2009 American Institute of Physics. [DOI: 10.1063/1.3099701]

Electrostatic turbulence driven by high magnetohydrodynamic activity in Tokamak Chauffage Alfven Bresilien

In Tokamak Chauffage Alfven Bresilien [R. M. O. Galvao , Plasma Phys. Controlled Fusion 43, 1181 (2001)], high magnetohydrodynamic (MHD) activity may appear spontaneously or during discharges with a voltage biased electrode inserted at the plasma edge. The turbulent electrostatic fluctuations, measured by Langmuir probes, are modulated by Mirnov oscillations presenting a dominant peak with a common frequency around 10 kHz. We report the occurrence of phase locking of the turbulent potential fluctuations driven by MHD activity at this frequency. Using wavelet cross-spectral analysis, we characterized the phase and frequency synchronization in the plasma edge region. We introduced an order parameter to characterize the radial dependence of the phase-locking intensity. (c) 2008 American Institute of Physics.

Deformed Gaussian-orthogonal-ensemble description of small-world networks

The study of spectral behavior of networks has gained enthusiasm over the last few years. In particular, random matrix theory (RMT) concepts have proven to be useful. In discussing transition from regular behavior to fully chaotic behavior it has been found that an extrapolation formula of the Brody type can be used. In the present paper we analyze the regular to chaotic behavior of small world (SW) networks using an extension of the Gaussian orthogonal ensemble. This RMT ensemble, coined the deformed Gaussian orthogonal ensemble (DGOE), supplies a natural foundation of the Brody formula. SW networks follow GOE statistics until a certain range of eigenvalue correlations depending upon the strength of random connections. We show that for these regimes of SW networks where spectral correlations do not follow GOE beyond a certain range, DGOE statistics models the correlations very well. The analysis performed in this paper proves the utility of the DGOE in network physics, as much as it has been useful in other physical systems.

Finding Quasi-Optimal Network Topologies for Information Transmission in Active Networks

This work clarifies the relation between network circuit (topology) and behaviour (information transmission and synchronization) in active networks, e.g. neural networks. As an application, we show how one can find network topologies that are able to transmit a large amount of information, possess a large number of communication channels, and are robust under large variations of the network coupling configuration. This theoretical approach is general and does not depend on the particular dynamic of the elements forming the network, since the network topology can be determined by finding a Laplacian matrix (the matrix that describes the connections and the coupling strengths among the elements) whose eigenvalues satisfy some special conditions. To illustrate our ideas and theoretical approaches, we use neural networks of electrically connected chaotic Hindmarsh-Rose neurons.

Soil nitrous oxide emissions in long-term cover crops-based rotations under subtropical climate

It has been shown that cover crops can enhance soil nitrous oxide (N(2)O) emissions, but the magnitude of increase depends on the quantity and quality of the crop residues. Therefore, this study aimed to evaluate the effect of long-term (19 and 21 years) no-till maize crop rotations including grass [black oat (Avena strigosa Schreb)] and legume cover crops [vetch (Vigna sativa L), cowpea (Vigna unguiculata L. Walp), pigeon pea (Cajanus cajan L. Millsp.) and lablab (Dolichos lablab)] on annual soil N(2)O emissions in a subtropical Acrisol in Southern Brazil. Greater soil N(2)O emissions were observed in the first 45 days after the cover crop residue management in all crop rotations, varying from -20.2 +/- 1.9 to 163.9 +/- 24.3 mu g N m(-2) h(-1). Legume-based crop rotations had the largest cumulative emissions in this period, which were directly related to the quantity of N (r(2) = 0.60, p = 0.13)and inversely related to the lignin:N ratio(r(2) = 0.89,p = 0.01) of the cover crop residues. After this period, the mean fluxes were smaller and were closely related to the total soil N stocks (r(2) = 0.96, p = 0.002). The annual soil N(2)O emission represented 0.39-0.75% of the total N added by the legume cover crops. Management-control led soil variables such as mineral N (NO(3)(-) and NH(4)(+)) and dissolved organic C influenced more the N(2)O fluxes than environmental-related variables as water-filled pore space and air and soil temperature. Consequently, the synchronization between N mineralization and N uptake by plants seems to be the main challenge to reduce N(2)O emissions while maintaining the environmental and agronomic services provided by legume cover crops in agricultural systems. (C) 2009 Elsevier B.V. All rights reserved.

Clinical relationships and ethical problems in primary care, Sao Paulo, SP, Brazil

Objectives: Main Objective: to identify ethical problems in primary care according to nurses` and doctors` perceptions. Secondary Objective: to know ethical issues of patient-professional relationships in primary care. Design: Synthesis to integrate and reinterpret primary results of qualitative studies. Setting: Primary healthcare centers, Sao Paulo, SP, Brazil. Participants and/or context: Incidental sample of 34 nurses and 36 medical doctors working in primary healthcare centers selected by convenience. Methods: Individual, semi-structured interviews to identity situations considered as sources of ethical problems. The sample is socially representative of primary care health centers and professionals. Data collection assured discourse saturation. Hermeneutic-dialectical discourse analysis was used to study the results. Results: Patient-professional relationships and team work were the main sources of ethical problems. The most important problems were patient information, privacy, confidentiality, interpersonal relationship, linkage and patient autonomy. These issues reflect the recent changes in clinical relation ships and show the peculiarities of primary care with its continuous care which lasts a long time. Healthcare involves multiprofessional team work in the midst of the patient claims for autonomy. Good care of patients needs requires a relationship based on communication and cooperation, and includes feelings and values, with communication skills. Conclusions: Ethical problems in primary care are common situations. For quality and humane primary care the relationship should consist of dialogue, trust and cooperation. (C) 2009 Elsevier Espana, S.L. All rights reserved.

The cultural adaptation and validation of the ""Burn Specific Health Scale-Revised"" (BSHS-R): Version for Brazilian burn victims

Background: The Burns Specific Health Scale-Revised (BSHS-R) is of easy application, can be self-administered, and it is considered a good scale to evaluate various important life aspects of burn victims. Objectives: To translate and culturally adapt the BSHS-R into the Brazilian-Portuguese language and to evaluate the internal consistency and convergent validity of the translated BSHS-R. Methods: The cultural adaptation of the BSHS-R included translation and back-translation, discussions with professionals and patients to ensure conceptual equivalence, semantic evaluation, and pre-test of the instrument. The Final Brazilian-Portuguese Version (FBPV) of the BSHS-R was tested on a group of 115 burn patients for internal consistency and validity of construct (using the Rosenberg Self-Esteem Scale (RSES) and the Beck Depression Inventory (BDI)). Results: All values of Cronbach`s alpha were greater than. 8, demonstrating that the internal consistency of the FBPV was very high. Self-esteem was highly correlated with affect and body image (r = .59, p < .001), and with interpersonal relationships (T = .51, p < .001). Correlations between the domains of the FBPV and the BDI were all negative but larger in magnitude than the correlations with RSES. Depression was highly correlated with affect and body image (r = -77, p < .001), and with interpersonal relationships (r = -67, p < .001). Conclusions: The results showed that the adapted version of the BSHS-R into Brazilian-Portuguese fulfills the validity and reliability criteria required from an instrument of health status assessment for burn patients. (C) 2008 Elsevier Ltd and ISBI. All rights reserved.

Cultural meaning of quality of life: perspectives of Brazilian burn patients

Purpose. This study was designed to explore the cultural meaning and dimensions of quality of life from the perspective of Brazilian burn patients. Method. A qualitative research approach was used. Nineteen burn patients and their close relatives participated in this ethnographic study. Data were collected by means of direct observation and semi-structured interviews, conducted in a hospital outpatient clinic and during visits to patients` homes. The following inter-related phases guided the analysis process: reading of the material and data reduction, data display, conclusion outlining and verification. Results. Participants reported that the quality of life is related to autonomy and the ability to work. The dimensions of quality of life included: resuming work and functional ability, body image, having leisure and interpersonal relationships. Their descriptions revealed their feelings and attitudes about resuming their previous activities and social lives, particularly concerning the work. Conclusion. For burn patients, quality of life is associated with the concept of normality, the satisfactory performance of social roles in the context of family life and the social world. The results showed the importance of the sociocultural dimension in the concept of quality of life for persons undergoing burn rehabilitation.

Chaotic communication based on the particle-in-a-box electronic circuit

A secure communication system based on the error-feedback synchronization of the electronic model of the particle-in-a-box system is proposed. This circuit allows a robust and simple electronic emulation of the mechanical behavior of the collisions of a particle inside a box, exhibiting rich chaotic behavior. The required nonlinearity to emulate the box walls is implemented in a simple way when compared with other analog electronic chaotic circuits. A master/slave synchronization of two circuits exhibiting a rich chaotic behavior demonstrates the potentiality of this system to secure communication. In this system, binary data stream information modulates the bifurcation parameter of the particle-in-a-box electronic circuit in the transmitter. In the receiver circuit, this parameter is estimated using Pecora-Carroll synchronization and error-feedback synchronization. The performance of the demodulation process is verified through the eye pattern technique applied on the recovered bit stream. During the demodulation process, the error-feedback synchronization presented better performance compared with the Pecora-Carroll synchronization. The application of the particle-in-a-box electronic circuit in a secure communication system is demonstrated.

Experimental study of a Tollmien-Schlichting wave interacting with a shallow 3D roughness element

The paper is devoted to an experimental study of the effect of a shallow 3D roughness element on the evolution of a 2D Tollmien-Schlichting wave in a Blasius boundary layer. The experiments were carried out under controlled disturbance conditions on an airfoil section which could provide a long run with zero pressure gradient flow. A pneumatically driven slit source was used to introduce the Tollmien-Schilichting wave upstream of the lower branch of the neutral stability curve. A few wavelengths downstream, the T-S wave interacts with a cylindrical roughness element. The height of the roughness was slowly oscillating in time, which allows a continuous measurement of the T-S wave response downstream the roughness. The oscillation frequency was approximately 1500 times lower than the frequency of the studied Tollmien-Schlichting wave and therefore, behaved as a steady roughness with respect to the T-S wave. Hot wire anemometry was used to measure wall normal profiles and spanwise scans close to the maximum of the eigenfunction of the T-S wave. The oscillation of the roughness and the synchronization of all-equipments permitted the use of ensemble average techniques. Two different amplitudes of T-S waves with a non-dimensional frequency of F120E-06 were studied. They show a strong amplification of the disturbances in a small spanwise wave number range. The analysis of the wall normal T-S profiles suggests the growth of oblique modes.

A 2-D Delaunay refinement algorithm using an initial prerefinement from the boundary mesh

An improvement to the quality bidimensional Delaunay mesh generation algorithm, which combines the mesh refinement algorithms strategy of Ruppert and Shewchuk is proposed in this research. The developed technique uses diametral lenses criterion, introduced by L. P. Chew, with the purpose of eliminating the extremely obtuse triangles in the boundary mesh. This method splits the boundary segment and obtains an initial prerefinement, and thus reducing the number of necessary iterations to generate a high quality sequential triangulation. Moreover, it decreases the intensity of the communication and synchronization between subdomains in parallel mesh refinement.