Use of phycoerythrin and allophycocyanin for fluorescence resonance energy transfer analyzed by flow cytometry: advantages and limitations.

BACKGROUND: This study validates the use of phycoerythrin (PE) and allophycocyanin (APC) for fluorescence energy transfer (FRET) analyzed by flow cytometry. METHODS: FRET was detected when a pair of antibody conjugates directed against two noncompetitive epitopes on the same CD8alpha chain was used. FRET was also detected between antibody conjugate pairs specific for the two chains of the heterodimeric alpha (4)beta(1) integrin. Similarly, the association of T-cell receptor (TCR) with a soluble antigen ligand was detected by FRET when anti-TCR antibody and MHC class I/peptide complexes (<<tetramers>>) were used. RESULTS: FRET efficiency was always less than 10%, probably because of steric effects associated with the size and structure of PE and APC. Some suggestions are given to take into account this and other effects (e.g., donor and acceptor concentrations) for a better interpretation of FRET results obtained with this pair of fluorochromes. CONCLUSIONS: We conclude that FRET assays can be carried out easily with commercially available antibodies and flow cytometers to study arrays of multimolecular complexes.

Embryonic gene expression of Coregonus palaea (whitefish) under pathogen stress as analyzed by high-throughput RNA-sequencing.

Most fishes produce free-living embryos that are exposed to environmental stressors immediately following fertilization, including pathogenic microorganisms. Initial immune protection of embryos involves the chorion, as a protective barrier, and maternally-allocated antimicrobial compounds. At later developmental stages, host-genetic effects influence susceptibility and tolerance, suggesting a direct interaction between embryo genes and pathogens. So far, only a few host genes could be identified that correlate with embryonic survival under pathogen stress in salmonids. Here, we utilized high-throughput RNA-sequencing in order to describe the transcriptional response of a non-model fish, the Alpine whitefish Coregonus palaea, to infection, both in terms of host genes that are likely manipulated by the pathogen, and those involved in an early putative immune response. Embryos were produced in vitro, raised individually, and exposed at the late-eyed stage to a virulent strain of the opportunistic fish pathogen Pseudomonas fluorescens. The pseudomonad increased embryonic mortality and affected gene expression substantially. For example, essential, upregulated metabolic pathways in embryos under pathogen stress included ion binding pathways, aminoacyl-tRNA-biosynthesis, and the production of arginine and proline, most probably mediated by the pathogen for its proliferation. Most prominently downregulated transcripts comprised the biosynthesis of unsaturated fatty acids, the citrate cycle, and various isoforms of b-cell transcription factors. These factors have been shown to play a significant role in host blood cell differentiation and renewal. With regard to specific immune functions, differentially expressed transcripts mapped to the complement cascade, MHC class I and II, TNF-alpha, and T-cell differentiation proteins. The results of this study reveal insights into how P. fluorescens impairs the development of whitefish embryos and set a foundation for future studies investigating host pathogen interactions in fish embryos.

Influence of carbon source and the fermentation process on levan production by Zymomonas mobilis analyzed by the surface response method

The aim of this study is to assess sugar cane juice and sucrose as substrates, the batch and fed batch processes and their interaction in the levan production using a complete factorial design. Zymomonas mobilis was cultivated in different sugar cane juice and sucrose concentrations in two fermentation processes at 25 °C for 20 h. A complete factorial design (2³) was used to analyze the effects of the type and concentration of the substrate, as well as the batch and fed batch processes. A complete second factorial design (2²) was used to observe the importance of sugar cane juice. The results indicated that the batch process improved the levan production reaching 40.14 g/L. The addition of sugar cane juice was not statistically significant for levan formation, however sugar cane juice stimulated biomass, sorbitol and ethanol production. The best medium for levan production was 150 g/L sucrose in batch.

Graphics for engineers, architects, and builders : a manual for designers, an a text - book for scientific schools. Trusses and arches analyzed and discussed by graphical methods

UANL

Graphics for engineers, architects, and builders : a manual for designers, and a text-book for technical school, trussesand arches analyzed and descussed by graphical methods

UANL

The development of organized structures in polyethylene crystallized from a sheared melt, analyzed by WAXS and TEM

The development of global orientation and morphological features in linear polyethylene crystallizing from a sheared melt are studied using in-situ time-resolving wide angle X-ray scattering (WAXS) and ex-situ transmission electron microscopy. It is found that samples subjected to a shear rate above a critical value of ~1s-1 result in macroscopically oriented structures in the crystallized sample. This critical shear rate appears to be independent of the differences in molecular weight distribution of the samples studied although the morphologies which develop are sensitive to quite small differences in molecular weight distributions. The presence of shish kebabs in the morphology is shown to differ markedly according to variations in the upper molecular weight fraction of the molecular weight distribution, even though the resulting global orientation does not. The WAXS also reveals that areas which evidence no row nucleated structures still realize high degrees of molecular orientation. It is proposed that the formation of shish kebab or lamellar morphologies in these samples is dependent on the critical density of contiguous elongated crystallization nuclei rather than any specific global criteria.

Characteristics of TMD noise analyzed by electrovibratography

Studying joint noise is an important parameter for diagnosing temporomandibular dysfunction. In this study, eight groups (n=9) were formed according to joint dysfunction classification, provided by employing vibration analysis equipment. Parameters for analyzing joint noise were: total vibration energy, peak amplitude, and peak frequency. Mouth opening range was also analyzed. Statistical analysis results for each parameter were significant at 1 %. Each analyzed group presented different noise characteristics. This allowed for inclusion of the groups within a determined value category. The patient group with normal condyle/disk relationship always presented the lowest values. The type of joint noise was characterized by analyzing total integral noise, peak amplitude, peak frequency, and mouth opening. Analyzing joint noise using electrovibratography suggests the type of joint dysfunction and may help to establish a diagnosis, as well as a treatment plan.

Mechanosensitivity of astrocytes on optimized polyacrylamide gels analyzed by quantitative morphometry

Cells are able to detect and respond to mechanical cues from their environment. Previous studies have investigated this mechanosensitivity on various cell types, including neural cells such as astrocytes. In this study, we have carefully optimized polyacrylamide gels, commonly used as compliant growth substrates, considering their homogeneity in surface topography, mechanical properties, and coating density, and identified several potential pitfalls for the purpose of mechanosensitivity studies. The resulting astrocyte response to growth on substrates with shear storage moduli of G` = 100 Pa and G` = 10 kPa was then evaluated as a function of coating density of poly-D-lysine using quantitative morphometric analysis. Astrocytes cultured on stiff substrates showed significantly increased perimeter, area, diameter, elongation, number of extremities and overall complexity if compared to those cultured on compliant substrates. A statistically significant difference in the overall morphological score was confirmed with an artificial intelligence-based shape analysis. The dependence of the cells` morphology on PDL coating density seemed to be weak compared to the effect of the substrate stiffness and was slightly biphasic, with a maximum at 10-100 mu g ml(-1) PDL concentration. Our finding suggests that the compliance of the surrounding tissue in vivo may influence astrocyte morphology and behavior.

Genetic parameters for milk yield analyzed by test-day models in Murrah buffaloes in Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Performance of aerodynamic baffles in cylindrical grinding analyzed on the basis of air layer pressure and speed

Over the years, grinding has been considered one of the most important manufacturing processes. Grinding is a high precision process, and the loss of a single workpiece in this stage of the production is unacceptable, fir the value added to the material is very high due to many processes it has already undergone prior to grinding. This study aims to contribute toward the development of an experimental methodology whereby the pressure and speed of the air layer produced by the high rotation of the grinding wheel is evaluated with and without baffles, i.e., in an optimized grinding operation and in a traditional one. Tests were also carried out with steel samples to check the difference in grinding wheel wear with and without the use of baffles.

Genotype by environment interaction for 450-day weight of Nelore cattle analyzed by reaction norm models

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Effect of fluid resins on the surface roughness and topography of resin composite restorations analyzed by atomic force microscope

The aim of the study was to verify the influence of surface sealants on the surface roughness of resin composite restorations before and after mechanical toothbrushing, and evaluate the superficial topography using atomic force microscope. Five surface sealers were used: Single Bond, Opti Bond Solo Plus, Fortify, Fortify Plus and control, without any sealer agent. The lowest values of surface roughness were obtained for control, Single Bond and Fortify groups before toothbrushing. Fortify and Fortify Plus were the sealer agents that support the abrasive action caused by the toothbrushing although Fortify Plus group remained with high values of surface roughness. The application of specific surface sealants could be a useful clinical procedure to maintain the quality of resin-based composite restorations. (C) 2010 Elsevier Ltd. All rights reserved.