• Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Current Issue in 2019

Volume: 9 Issue: 17

Biochemistry

Photoaffinity Labeling of Respiratory Complex I in Bovine Heart Submitochondrial Particles by Photoreactive [125I] amilorides Authors:  Masatoshi Murai and Hideto Miyoshi, date: 09/05/2019, view: 131, Q&A: 0
The architecture of quinone/inhibitor-access channel in proton-translocating NADH-quinone oxidoreductase (respiratory complex I) was modeled by X-ray crystallography and cryo-EM, however, it remains debatable whether the channel model reflects the physiologically relevant state present throughout the catalytic cycle. Using photoreactive [125 ...
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Cancer Biology

A Robust, One-step FRET Assay for Human Heparanase Authors:  Jyothi C. Sistla and Umesh R. Desai, date: 09/05/2019, view: 159, Q&A: 0
Heparanase, an endo-β-D-glucuronidase, cleaves cell surface and extracellular matrix heparan sulfate (HS) chains at distinct sites and plays important biological roles including modulation of cell growth and metastasis. Although a number of different types of heparanase assays have been reported to date, most are labor intensive, complex ...
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Cell Biology

In vitro Intestinal Epithelial Wound-healing Assays Using Electric Cell-Substrate Impedance Sensing Instrument Authors:  Olivier Merlin-Zhang, Junsik Sung and Emilie Viennois, date: 09/05/2019, view: 200, Q&A: 0
Here, we describe an in vitro epithelial wound-healing assay using Electric Cell-Substrate Impedance Sensing (ECIS) technology. The ECIS technology is a real time cell growth assay based on a small (250 μm diameter) active gold electrode which resistance is measured continuously. When intestinal epithelial cells reach confluency on the ...
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Developmental Biology

Estrous Cycle Monitoring in Mice with Rapid Data Visualization and Analysis Authors:  Leanna K. Pantier, Jiang Li and Catherine A. Christian, date: 09/05/2019, view: 312, Q&A: 0
The estrous cycle provides a readout of reproductive health in female laboratory rodents, and estrous cycle stage can be an important physiological variable. Accurate assessment of estrous cycle stage is also important in producing timed pregnancies for developmental studies. Here, we provide a protocol for evaluation of estrous cycle stage ...
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A Widely Applicable Urea-based Fluorescent/Colorimetric mRNA in situ Hybridization Protocol Author:  Chiara Sinigaglia, date: 09/05/2019, view: 226, Q&A: 0
In situ hybridization methods are routinely employed to detect nucleic acid sequences, allowing to localize gene expression or to study chromosomal organization in their native context. These methods rely on the pairwise binding of a labeled probe to the target endogenous nucleic acid sequence–the hybridization step, followed by detection ...
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Immunology

A Refined Protocol for Identifying Citrulline-specific Monoclonal Antibodies from Single Human B Cells from Rheumatoid Arthritis Patient Material Authors:  Khaled Amara, Lena Israelsson, Ragnhild Stålesen, Peter Sahlström, Johanna Steen, Vivianne Malmström and Caroline Grönwall, date: 09/05/2019, view: 149, Q&A: 0
We describe here a detailed, refined protocol for the generation of citrulline-specific monoclonal antibodies from single human B cells from rheumatoid arthritis (RA) patients. This protocol provides a detailed guide of the procedure starting from single B cells of your choice and followed by amplification of the variable region of immunoglobulin ...
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Isolation of Neutrophil Nuclei for Use in NETosis Assays Author:  Dave Boucher, date: 09/05/2019, view: 262, Q&A: 0
Neutrophils are critical immune cells that protect our body against invading pathogens. They generate antibacterial DNA structures called neutrophil extracellular traps (NET). Recently we identified a new mechanism that enables NET formation. We observed that following recognition of lipopolysaccharides, inflammatory caspases cleave Gasdermin D ...
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Microbiology

Development of an Efficient Transformation System for Halotolerant Yeast Debaryomyces hansenii CBS767 Authors:  Anu P. Minhas and Dipanwita Biswas, date: 09/05/2019, view: 137, Q&A: 0
Debaryomyces hansenii is one of the most osmotolerant and halotolerant yeasts. Further, its association with traditional cheese and meat products imparting special flavors to these products project this yeast with enormous biotechnological potential in the agrofood sector. However, lack of an efficient transformation system in D. hansenii ...
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Yeast Single-cell RNA-seq, Cell by Cell and Step by Step Authors:  Mariona Nadal-Ribelles, Saiful Islam, Wu Wei, Pablo Latorre, Michelle Nguyen, Eulàlia de Nadal, Francesc Posas and Lars M. Steinmetz, date: 09/05/2019, view: 197, Q&A: 0
Single-cell RNA-seq (scRNA-seq) has become an established method for uncovering the intrinsic complexity within populations. Even within seemingly homogenous populations of isogenic yeast cells, there is a high degree of heterogeneity that originates from a compact and pervasively transcribed genome. Research with microorganisms such as yeast ...
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High Resolution Respirometry in Candida albicans Authors:  Lucian Duvenage, Carol A. Munro and Campbell W. Gourlay, date: 09/05/2019, view: 123, Q&A: 0
Many Candida species, such as the opportunistic human pathogen Candida albicans, are Crabtree-Negative yeasts and are therefore highly dependent on the energy generated through oxidative phosphorylation. Respiration control is linked to a range of aspects of C. albicans cell physiology that appear to be important for ...
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Neuroscience

A Novel Dual Lentiviral CRISPR-based Transcriptional Activation System for Gene Expression Regulation in Neurons Authors:  Katherine E. Savell, Faraz A. Sultan and Jeremy J. Day, date: 09/05/2019, view: 513, Q&A: 0
Robust and efficient gene expression control enables the study of a gene’s function in the central nervous system. Advances in CRISPR-based technology provide new avenues not only for gene editing, but for complex transcriptional control. Here, we describe a protocol to generate high-titer lentiviruses with neuron-optimized CRISPR-activation ...
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Co-culture of Human Stem Cell Derived Neurons and Oligodendrocyte Progenitor Cells Authors:  Stephanie Dooves, Aishwarya G. Nadadhur, Lisa Gasparotto and Vivi M. Heine, date: 09/05/2019, view: 267, Q&A: 0
Crosstalk between neurons and oligodendrocytes is important for proper brain functioning. Multiple co-culture methods have been developed to study oligodendrocyte maturation, myelination or the effect of oligodendrocytes on neurons. However, most of these methods contain cells derived from animal models. In the current protocol, we co-culture ...
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A High-throughput and Pathophysiologically Relevant Astrocyte-motor Neuron Co-culture Assay for Amyotrophic Lateral Sclerosis Therapeutic Discovery Authors:  Matthew J. Stopford, Scott P. Allen and Laura Ferraiuolo, date: 09/05/2019, view: 392, Q&A: 0
Amyotrophic lateral sclerosis (ALS) is an adult onset neurological disorder characterized by loss of motor neurons leading to progressive muscle wasting and eventually death. Astrocytes play a key role in disease pathogenesis. However, the ability to study astrocytic support towards motor neurons in ALS has been limited by a lack of sustainable ...
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Plant Science

PEG-mediated, Stable, Nuclear and Chloroplast Transformation of Cyanidioschizon merolae Authors:  Maksymilian Zienkiewicz, Tomasz Krupnik, Anna Drożak and Kinga Kania, date: 09/05/2019, view: 122, Q&A: 0
The ability to achieve nuclear or chloroplast transformation in plants has been a long standing goal, especially in microalgae research. Over past years there has been only little success, but transient and stable nuclear transformation has been achieved in multiple species. Our newly developed method allows for relatively simple transformation of ...
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