• Volume 11, 2021
  • Volume 10, 2020
  • 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
Image Credit: BioProtoc.3987

Past Issues in 2021

Volume: 11 Issue: 8

Biochemistry

Quantitative Characterization of the Amount and Length of (1,3)-β-D-glucan for Functional and Mechanistic Analysis of Fungal (1,3)-β-D-glucan Synthase Authors:  Abhishek Chhetri, Anna Loksztejn and Kenichi Yokoyama, date: 04/20/2021, view: 713, Q&A: 0

(1,3)-β-d-Glucan synthase (GS) is an essential enzyme for fungal cell wall biosynthesis that catalyzes the synthesis of (1,3)-β-d-glucan, a major and vital component of the cell wall. GS is a proven target of antifungal antibiotics including FDA-approved echinocandin derivatives; however, the function and mechanism of GS remain largely

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Optimized Recombinant Production of Secreted Proteins Using Human Embryonic Kidney (HEK293) Cells Grown in Suspension Authors:  Silvia Faravelli, Matteo Campioni, Martina Palamini, Anselmo Canciani, Antonella Chiapparino and Federico Forneris, date: 04/20/2021, view: 896, Q&A: 0

Recombinant proteins are an essential milestone for a plethora of different applications ranging from pharmaceutical to clinical, and mammalian cell lines are among the currently preferred systems to obtain large amounts of proteins of interest due to their high level of post-translational modification and manageable large-scale production. In

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Cancer Biology

A Robust Mammary Organoid System to Model Lactation and Involution-like Processes Authors:  Elsa Charifou, Jakub Sumbal, Zuzana Koledova, Han Li and Aurélie Chiche, date: 04/20/2021, view: 721, Q&A: 0

The mammary gland is a highly dynamic tissue that changes throughout reproductive life, including growth during puberty and repetitive cycles of pregnancy and involution. Mammary gland tumors represent the most common cancer diagnosed in women worldwide. Studying the regulatory mechanisms of mammary gland development is essential for understanding

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Cell Biology

AIMTOR, a BRET Biosensor for Live Recording of mTOR Activity in Cell Populations and Single Cells Authors:  Nathalie Bouquier, Enora Moutin, Julie Perroy and Vincent Ollendorff, date: 04/20/2021, view: 663, Q&A: 0

Mammalian target of rapamycin (mTOR) controls many crucial cellular functions, including protein synthesis, cell size, energy metabolism, lysosome and mitochondria biogenesis, and autophagy. Consequently, deregulation of mTOR signaling plays a role in numerous pathological conditions such as cancer, metabolic disorders and neurological diseases.

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Developmental Biology

Production of Phenotypically Uniform Human Cerebral Organoids from Pluripotent Stem Cells Authors:  Adam Sivitilli, Parisa Ghiasi and Liliana Attisano, date: 04/20/2021, view: 609, Q&A: 0

Recent advances in stem cell technology have allowed researchers to generate 3D cerebral organoids (COs) from human pluripotent stem cells (hPSCs). Indeed, COs have provided an unprecedented opportunity to model the developing human brain in a 3D context, and in turn, are suitable for addressing complex neurological questions by leveraging

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Development of a Chemical Reproductive Aging Model in Female Rats Authors:  Nayara Pestana-Oliveira, Ruither O. G. Carolino, Bruna Kalil-Cutti, Cristiane M. Leite, Litamara C. Dalpogeto, Bruna Balbino De Paula, Jonh P. Collister and Janete Anselmo-Franci, date: 04/20/2021, view: 703, Q&A: 0

Women are born with an abundant but finite pool of ovarian follicles, which naturally and progressively decreased during their reproductive years until menstrual periods stop permanently (menopause). Perimenopause represents the transition from reproductive to non-reproductive life. It is usually characterized by neuroendocrine, metabolic and

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Microbiology

A Sensitive and Specific PCR-based Assay to Quantify Hepatitis B Virus Covalently Closed Circular (ccc) DNA while Preserving Cellular DNA Authors:  Benno Zehnder, Stephan Urban and Thomas Tu, date: 04/20/2021, view: 519, Q&A: 0

Hepatitis B virus (HBV) is the major cause of liver diseases and liver cancer worldwide. After infecting hepatocytes, the virus establishes a stable episome (covalently closed circular DNA, or cccDNA) that serves as the template for all viral transcripts. Specific and accurate quantification of cccDNA is difficult because infected cells contain

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Measuring Cytosolic Translocation of Mycobacterium marinum in RAW264.7 Macrophages with a CCF4-AM FRET Assay Authors:  Javier Aguilera and Jianjun Sun, date: 04/20/2021, view: 459, Q&A: 0
The CCF4-AM Förster resonance energy transfer (FRET) assay is a sensitive approach to measure bacterial cytosolic translocation in live cells. The FRET pair hydroxycoumarin (donor) and fluorescein (acceptor) are linked by a CCF4-AM β-lactam ring, the substrate of β-lactamase. The exogenously added, neutral charged-FRET reagent can diffuse across ...
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Single Cell Analysis and Sorting of Aspergillus fumigatus by Flow Cytometry Authors:  Gareth Howell and Robert-Jan Bleichrodt, date: 04/20/2021, view: 534, Q&A: 0

Experimental results in fungal biology research are usually obtained as average measurements across whole populations of cells, whilst ignoring what is happening at the single cell level. Microscopy has allowed us to study single-cell behavior, but it has low throughput and cannot be used to select individual cells for downstream experiments. Here

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Screening for Lysogen Activity in Therapeutically Relevant Bacteriophages Authors:  Fernando L. Gordillo Altamirano and Jeremy J. Barr, date: 04/20/2021, view: 754, Q&A: 2

Lysogenic phages can integrate into their bacterial host’s genome, potentially transferring any genetic information they possess including virulence or resistance genes, and are therefore routinely excluded from therapeutic applications. Lysogenic behavior is typically seen in phages that create turbid plaques or possess subpar bactericidal

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Neuroscience

A Method to Quantify Drosophila Behavioral Activities Induced by GABAA Agonist Muscimol Author:  M. Rezaul Karim, date: 04/20/2021, view: 358, Q&A: 0

Muscimol is a psychoactive isoxazole derived from the mushroom Amanita muscaria. As a potent GABAA receptor agonist, muscimol suppresses the activity of the central nervous system, reduces anxiety and induces sleep. We investigated the effects of muscimol on Drosophila behavior. Drosophila behavioral assays are powerful tools that are used to

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Single or Repeated Ablation of Mouse Olfactory Epithelium by Methimazole Authors:  Sofia Håglin, Staffan Bohm and Anna Berghard, date: 04/20/2021, view: 410, Q&A: 0

Odor-detecting olfactory sensory neurons residing in the nasal olfactory epithelium (OE) are the only neurons in direct contact with the external environment. As a result, these neurons are subjected to chemical, physical, and infectious insults, which may be the underlying reason why neurogenesis occurs in the OE of adult mammals. This feature

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Single-unit Recording in Awake Behaving Non-human Primates Authors:  Mengxi Yun, Masafumi Nejime and Masayuki Matsumoto, date: 04/20/2021, view: 401, Q&A: 0

Non-human primates (NHPs) have been widely used as a species model in studies to understand higher brain functions in health and disease. These studies employ specifically designed behavioral tasks in which animal behavior is well-controlled, and record neuronal activity at high spatial and temporal resolutions while animals are performing the

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Transfection and Activation of CofActor, a Light and Stress Gated Optogenetic Tool, in Primary Hippocampal Neuron Cultures Authors:  Wyatt P. Bunner, Rachel Dodson, Robert M. Hughes and Erzsebet M. Szatmari, date: 04/20/2021, view: 451, Q&A: 0

Proteins involved in neurodegeneration can be coupled with optogenetic reagents to create rapid and sensitive reporters to provide insight into the biochemical processes that mediate the progression of neurodegenerative disorders, including Alzheimer’s Disease (AD). We have recently developed a novel optically-responsive tool (the

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Systems Biology

Computational Analysis and Phylogenetic Clustering of SARS-CoV-2 Genomes Authors:  Bani Jolly and Vinod Scaria, date: 04/20/2021, view: 786, Q&A: 0

COVID-19, the disease caused by the novel SARS-CoV-2 coronavirus, originated as an isolated outbreak in the Hubei province of China but soon created a global pandemic and is now a major threat to healthcare systems worldwide. Following the rapid human-to-human transmission of the infection, institutes around the world have made efforts to generate

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