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Past Issues in 2015

Volume: 5 Issue: 5

Immunology

Isolation of Splenic Dendritic Cells Using Fluorescence-activated Cell Sorting Authors: Simon J Tavernier, Fabiola Osorio, Sophie Janssens and Bart N Lambrecht, date: 03/05/2015, view: 13621, Q&A: 0

The spleen is a vastly vasculated organ and consists of a complex organized network of innate and adaptive immune cells. This permits the specialized functions of the spleen such as antibacterial and antifungal immunity and iron metabolism among others (Mebius and Kraal, 2005). Different dendritic cell (DC) subsets reside in the spleen and can be ...

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Microbiology

Differentiation of Naturally Produced Extracellular Membrane Vesicles from Lipid Aggregation by Glucuronoxylomannan Immunogold Transmission Electron Microscopy in Bacillus subtilis Authors: Lisa Brown, Geoff Perumal and Arturo Casadevall, date: 03/05/2015, view: 8158, Q&A: 0

Recently, membrane vesicle (MV) production was described in Gram-positive bacteria, which harbor a variety of components such as toxins, antibiotic resistance proteins, proteases, DNA, and immune modulators. Free lipids have the ability to form micelles, thus it is important to rule out spontaneous association of lipids into vesicle-like ...

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Vesicle Isolation from Bacillus subtilis Biofilm Authors: Lisa Brown, Anne Kessler and Arturo Casadevall, date: 03/05/2015, view: 10462, Q&A: 0

Bacterial biofilms are associated clinically with many bacterial infections including those caused by bacteria such as Pseudomonas aeruginosa and Staphylococcus aureus. In recent years, extracellular vesicles produced by bacteria have been isolated from biofilm communities. Vesicles have been described in depth and can ...

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A Gas Chromatography-Mass Spectrometry-Based Two Stage Assay for Measurement of in vitro myo-Inositol 3-phosphate Synthase (INO1) Activity Authors: James I. MacRae and Malcolm J. McConville, date: 03/05/2015, view: 8031, Q&A: 0

This method describes an in vitro assay for measuring INO1 enzyme activity (the conversion of glucose 6-phosphate to myo-inositol 3-phosphate) in cell-free extracts. The method was first described for Plasmodium falciparum cells in MacRae et al. (2014) and consists of two parts: Part 1 describes the assay itself ...

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Neuroscience

Thirty-Second Net Stressor Task in Adult Zebrafish Authors: Steven Tran and Robert Gerlai, date: 03/05/2015, view: 7320, Q&A: 0

Zebrafish have become a popular animal model for behavioral neuroscience (Gerlai, 2014). Recent studies have demonstrated that brief experimental handling prior to euthanizing animals can subsequently alter biological measures quantified post-mortem (e.g. cortisol levels) (Ramsay et al., 2009; Tran et al., 2014). Here we ...

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Plant Science

RNA Editing Detection by Direct Sequencing Authors: Mehdi Jabnoune, David Secco, Cécile Lecampion, Christophe Robaglia, Qingyao Shu and Yves Poirier, date: 03/05/2015, view: 7239, Q&A: 0

RNA editing is a widespread post-transcriptional phenomenon through which primary RNA sequences are altered by nucleotide insertion/deletion or base conversion. It occurs in a variety of organisms and cooperates with alternative splicing in increasing both proteomic and transcriptomic complexity. We describe here a method allowing RNA editing ...

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Isolation of Polysome-bound mRNA from Rice Solid Tissues Amenable for RT-PCR and Profiling Experiments Authors: Mehdi Jabnoune, David Secco, Cécile Lecampion, Christophe Robaglia, Qingyao Shu and Yves Poirier, date: 03/05/2015, view: 11568, Q&A: 0

Polysome profile analysis is a frequently performed task in translational control research that not only enables direct monitoring of the efficiency of translation but can easily be extended with a wide range of downstream applications such as Northern and western blotting, genome-wide microarray analysis or qRT-PCR. Here, we describe a method for ...

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An Efficient Procedure for Protoplast Isolation from Mesophyll Cells and Nuclear Fractionation in Rice Authors: Mehdi Jabnoune, David Secco, Cécile Lecampion, Christophe Robaglia, Qingyao Shu and Yves Poirier, date: 03/05/2015, view: 19259, Q&A: 1

Plant protoplasts, a proven physiological and versatile cell system, are widely used in high-throughput analysis and functional characterization of genes. Green protoplasts have been successfully used in investigations of plant signal transduction pathways related to hormones, metabolites and environmental challenges. This protocol, adapted from ...

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Measurement of Cellular Redox in Pollen with Redox-Sensitive GFP (roGFP) Using Live Cell Imaging Authors: Wei-Jie Huang and Wei-Hua Tang, date: 03/05/2015, view: 8318, Q&A: 0

Redox homeostasis is a fundamental property of living cells and responds actively to both cellular metabolism and external stimulus. The development of redox-sensitive GFP (roGFP) enables dynamic monitoring of changes in cellular redox poise (Hanson et al., 2014). When excited alternatively at 405 nm and 488 nm, these probes exhibit ...

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Citrus Fruit Ascorbic Acid Extraction and Quantification by HPLC Authors: Enriqueta Alós, Joanna Lado, María Jesús Rodrigo and Lorenzo Zacarías, date: 03/05/2015, view: 15834, Q&A: 0

Citrus are among the most relevant sources of vitamin C (ascorbic acid + dehydroascorbic acid). Recent studies have revealed that it increases in the peel as fruit ripens and remains constant or even decreases in the pulp tissue. Moreover, important differences on ascorbic acid content exist among citrus varieties in both tissues. Here we describe ...

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Extraction of Small RNA and qPCR Validation of miRNAs in Vigna mungo Authors: Sujay Paul, Anirban Kundu and Amita Pal, date: 03/05/2015, view: 8680, Q&A: 0

Small RNAs like microRNAs (miRNAs), small interfering RNAs (siRNAs) and other noncoding RNAs including snRNA and snoRNA have tremendous impact on eukaryotic gene regulation. Extraction of high quality small RNAs is an important prerequisite for experimental analyses of miRNAs. This will prevent RNA degradation and remove associated contaminations ...

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