Sensitivity of CFP/YFP and GFP/mCherry pairs to donor photobleaching on FRET determination by fluorescence lifetime imaging microscopy in living cells

Fluorescent protein-based FRET is a powerful method for visualizing protein-protein interactions and biochemical reactions in living cells. It can be difficult, however, to avoid photobleaching when observing fluorescent cells under the microscope, especially those expressing CFP. We compared the sensitivity of two protein-based FRET pairs to light-induced fluorescence changes in the donor, on FRET determination by fluorescence lifetime imaging microscopy (FLIM). Thanks to the very low excitation light levels of the time- and space-correlated single photon counting (TSCSPC) method, FLIM acquisitions were achieved without donor photobleaching. Here, we show that photobleaching of CFP by a mercury lamp under the microscope induced a decrease in the mean fluorescence lifetime, which interfered with FRET determination between CFP and YFP. Importantly, the range of light-induced variation of the mean fluorescence lifetime of CFP was not proportional to the decrease in the steady state fluorescence intensity and varied from cell to cell. The choice of the CFP/YFP pair therefore requires that the cells be observed and analyzed at very low light levels during the whole FRET experiment. In contrast, the GFP/mCherry pair provided an accurate FRET measurement by FLIM, even if some GFP photobleaching took place. We thus demonstrate that CFP can be an unreliable donor for FRET determination in living cells, due to its photosensitivity properties. We demonstrate that the GFP/mCherry pair is better suited for FRET measurement by FLIM in living cells than the CFP/YFP pair.     

Cysteine tagging for MS-based proteomics

Amino acid-tagging strategies are widespread in proteomics. Because of the central role of mass spectrometry (MS) as a detection technique in protein sciences, the term "mass tagging" was coined to describe the attachment of a label, which serves MS analysis and/or adds analytical value to the measurements. These so-called mass tags can be used for separation, enrichment, detection, and quantitation of peptides and proteins. In this context, cysteine is a frequent target for modifications because the thiol function can react specifically by nucleophilic substitution or addition. Furthermore, cysteines present natural modifications of biological importance and a low occurrence in the proteome that justify the development of strategies to specifically target them in peptides or proteins. In the present review, the mass-tagging methods directed to cysteine residues are comprehensively discussed, and the advantages and drawbacks of these strategies are addressed. Some concrete applications are given to underline the relevance of cysteine-tagging techniques for MS-based proteomics.     

Study of the impact of a new hurdle technology composed of electro‐activated solution and low heat treatment on the canned pea and corn quality and microbial safety

The objective of this work was to evaluate the quality of sterilised canned pea and corn in electro‐activated brine solutions at moderate temperatures. The lowest change in vitamin C was associated with the lowest heat treatment, while the short treatment time resulted in significant changes in texture and colour of vegetables. Best texture profile was obtained with the acid electro‐activated brine solution for pea and corn. The neutral electro‐activated brine solution resulted in a less firm texture for pea and corn. The green colour and brightness of canned pea were higher when neutral solution was used (a* = ?8.4 ± 0.3) than for the acidic one (a* = ?3.7 ± 0.6). The yellowness of corn was better with the neutral electro‐activated brine solution (b* = 36.32 ± 1.24) than with the acidic one (b* = 28.44 ± 2.39). Thirty‐three percent decrease of energy consumption was obtained using the electro‐activation technology.     

Bidirectional HMM-based Arabic POS tagging

In this work, we will present a new concept of POS tagging that will be implemented for the Arabic language. Indeed, we will see that in Arabic there are a numerous cases where the determination of the morpho-syntactic state of a word depends on the states of the subsequent words, which represents the theoretical foundation of the approach: how to consider, in addition of the past elements, the future ones. We will then demonstrate how the POS tagging in its statistical application: the HMM, is based mainly on the past elements, and how to combine both direct and reverse taggers to tag the same sequence of words in both senses. Thus, we will propose a hypothesis for the result selecting. In the practical part, we will present, in general, the used resource and the changes made on it. Then we will explain the experiment steps and the parameters collected and presented on graphics, that we will discuss later to lead to the final conclusion.     

Gaussian beam conversion using an axicon

Abstract

The paper presents a theoretical investigation of laser beam tailoring using a transparent diffracting phase plate. It is shown that a linear axicon is able to transform a Gaussian beam either into a super-Gaussian profile, a ring-shaped profile or a doughnut profile. All these different intensity distributions are easily obtained by only changing the width of the incident Gaussian beam.     

Moment-Based Angular Difference Estimation Between Two Tomographic Projections in 2D and 3D

This paper introduces a new method for estimating the angular difference between two tomographic projections belonging to a set of projections taken at unknown directions in 2D and 3D. Our method relies on the projection neighbor selection in projection moment space, the calculation of the angular differences between these neighboring projections using moment properties and a projection moment neighborhood graph. The accuracy and the robustness of our method are shown on a test database including fifty 2D and 3D gray-level images at different resolutions and with different levels of noise.     

Development of the Analysis of Fecal Stanols in the Oyster Crassostrea gigas and Identification of Fecal Contamination in Shellfish Harvesting Areas

The objective of this work was to study the effects of washing and purification steps on qualitative and quantitative analysis of fecal stanols in the oyster Crassostrea gigas using either single or a combination of lipid purification steps on silica gel or aminopropyl bonded silica gel (NH₂) or a washing step. Among the three analytical pathways compared, the two including water extraction or NH₂ purification did not lead to higher recoveries and decreased repeatabilities of extractions compared to the single purification on silica gel. This latter led to similar recoveries (ca. 80 %) and repeatabilities (ca. 10 %) for both spiked standards (coprostanol and sitostanol). This analytical pathway has been applied to oysters collected in a harvesting area in Brittany (France) where fecal contaminations are important and allowed to quantify eight stanols in oysters. The relative proportions of fecal stanols of these oysters were combined with principal component analysis in order to investigate the usefulness of their stanol fingerprints to record a fecal contamination and to distinguish its source between human, porcine and bovine contaminations. Oysters non-fecally contaminated by Escherichia coli did not present specific stanol fingerprints while oysters fecally contaminated had a bovine fingerprint, suggesting a contamination of these samples by bovine sources. As a consequence, the method developed here allows the use of stanol fingerprints of oysters as a microbial source tracking tool that can be applied to shellfish harvesting areas subjected to fecal contaminations in order to identify the different sources of contamination and improve watershed management.     

Evaluation of ascorbic acid and sugar degradation products during fruit dessert processing under conventional or ohmic heating treatment

The effects of conventional heating (C) or ohmic heating (O) treatment on the degradation of sugar and ascorbic acid (AA) were evaluated in chunky fruit desserts prepared with apple puree (Golden Delicious var.) and chunky peach (Panavi var.) pieces. Concentrations of 5-HMF, furfural (F), 3-hydroxy-2-pyrone (3H2P) and 2-furoic acid (FA) were determined to assess the importance of oxidative and/or thermal reactions during processing. Depending on the thermal processing, 5-HMF and F levels in samples ranged from not detected to 3260 and from not detected to 570 μg/100 g FW respectively. Production of 3H2P and FA ranged from 480 to 2670 and from 84 to 420 μg/100 g FW respectively. Results showed that the impact of final heating treatment on the formation of AA degradation products (3H2P and FA) was minor, if any. Thus, AA degradation during apple products processing mainly depended on oxidative reactions pathway. Likewise, ohmic heating processing had very little effect on the formation of 5-HMF and F, in opposite to conventional heating.     

Physiological and biochemical markers in the process of resistance and/or tolerance of heavy metals in the abandoned mining area of Sidi Kamber,Skikda, Algeria

This paper reports the biomonitoring of metallic pollution in the mine of Sidi Kamber Skikda town, to determine the resistance and/or tolerance to Zn, Pb, Cd and Cu in metalliferous plants. During the period of March–May, 2015, soil and plants samples were collected from three different stations in the study area. The total heavy metal fraction was determined by ICP/MS, whereas biochemical markers were determined by spectrophotometric techniques. The results obtained showed high levels of heavy metals in soil and plants compared to international standards. The translocation factor revealed that the studied plants can transfer and/or accumulate metals in their roots or their aerial parts as a function of characteristics of each metal, soil contents and physiological role in the plant. The contents of chlorophyll, proline, total sugars and total proteins are directly influenced by changes in bioavailable contents of heavy metals. It has been found that the plants studied and biomarkers are better suited for phytoremediation and biomonitoring of heavy metals pollution.     

A Multiphase Pore Scale Network Model of Gas Exchange in Apple Fruit

A multiphase pore scale network model was developed to describe mass transfer in apple fruit. The 3D microscale geometry of the tissue was reconstructed from synchrotron radiation tomography images. Individual cells and pores were identified using a watershed segmentation procedure on a Euclidean distance map of the tissue microstructure. Further morphological characteristics of each individual pore, including its volume, connections to the neighbors and the connected area between the pore and its neighbors, were determined. The tissue was represented by a network of nodes (simplified individual pores and cells) that were interconnected by tubes. The transport of the respiratory gases O₂ and CO₂ between two nodes was modelled using diffusion laws and irreversible thermodynamics, while respiration was taken into account in the individual cellular nodes. A numerical procedure was applied to simulate the gas transport within the discrete network and to compute the local diffusivities of the links in the network. The predicted overall gas diffusivities compared well to experimental data and results computed from a microscale continuum model, thereby validating the pore scale network model. This approach is a computationally attractive alternative to a continuum multiphase approach for modelling gas transport in fruit.