• 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

Biochemistry

Proximity Ligation Assay for the Investigation of the Intramolecular Interaction of ELMO1 Authors:  Wai Wa Ray Chan, Dik Long Dennis Chau, Wen Li and Kwok-Fai Lau, date: 12/05/2019, view: 59, Q&A: 0
Intramolecular interaction is a common mechanism that regulates protein activities. Conventionally, such interactions are investigated by classical in vitro biochemical assays. Here, we describe a protocol for studying the intramolecular interaction of cell motility and engulfment 1 (ELMO1) in mammalian cells by using proximity ligation ...
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Cancer Biology

An Optimized Method to Isolate Human Fibroblasts from Tissue for Ex Vivo Analysis Authors:  Sara Waise, Rachel Parker, Matthew J. J. Rose-Zerilli, David M. Layfield, Oliver Wood, Jonathan West, Christian H. Ottensmeier, Gareth J. Thomas and Christopher J. Hanley, date: 12/05/2019, view: 49, Q&A: 0
Despite their involvement in many physiological and pathological processes, fibroblasts remain a poorly-characterized cell type. Analysis of primary fibroblasts while maintaining their in vivo phenotype is challenging: standard methods for fibroblast isolation require cell culture in vitro, which is known to alter phenotypes. ...
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DNase I Chromatin Accessibility Analysis Authors:  Brook S. Nepon-Sixt and Mark G. Alexandrow, date: 12/05/2019, view: 48, Q&A: 0
Chromatin consists of compacted DNA in complex with proteins and contributes to the organization of DNA and its stability. Furthermore, chromatin plays key roles in regulating cellular processes such as DNA replication, transcription, DNA repair, and mitosis. Chromatin assumes more compact (inaccessible) or decondensed (accessible) conformations ...
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Enhanced-ice-COLD-PCR for the Sensitive Detection of Rare DNA Methylation Patterns in Liquid Biopsies Authors:  Florence Mauger and Jörg Tost, date: 12/05/2019, view: 41, Q&A: 0
In the context of precision medicine, the identification of novel biomarkers for the diagnosis of disease, prognosis, predicting treatment outcome and monitoring of treatment success is of great importance. The analysis of methylated circulating-cell free DNA provides great promise to complement or replace genetic markers for these applications, ...
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Cell Biology

Quantification of Mitochondrial Dynamics in Fission Yeast Authors:  Leeba Ann Chacko and Vaishnavi Ananthanarayanan, date: 12/05/2019, view: 50, Q&A: 0
Mitochondria are double-membraned organelles responsible for several functions in the cell including energy production, calcium signaling, and cellular metabolism. An equilibrium between fission and fusion events of mitochondria is required for their proper functioning. Mitochondrial morphologies have been quantified in yeast using image ...
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Microbiology

Herpes Simplex Virus Type 1 Propagation, Titration and Single-step Growth Curves Authors:  Linda Grosche, Katinka Döhner, Alexandra Düthorn, Ana Hickford-Martinez, Alexander Steinkasserer and Beate Sodeik, date: 12/05/2019, view: 52, Q&A: 0
Given the endemic seroprevalence of herpes simplex viruses (HSV), its associated human diseases, and the emergence of acyclovir-resistant strains, there is a continuous need for better antiviral therapies. Towards this aim, identifying mechanistic details of how HSV-1 manipulates infected cells, how it modulates the immune responses, and how it ...
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Quantification of Nitric Oxide and Reactive Oxygen Species in Leishmania-infected J774A.1 Macrophages as a Response to the in vitro Treatment with a Natural Product Compound Authors:  Kalliopi Karampetsou, Olga S. Koutsoni and Eleni Dotsika, date: 12/05/2019, view: 40, Q&A: 0
Leishmaniasis is a parasitic disease caused by the obligatory intracellular protozoa Leishmania spp. Current therapeutic options are limited and thus, drug discovery against leishmaniasis is very important. Nevertheless, there is a great difficulty to develop therapeutic strategies against the disease because the parasite deploys various ...
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Detection of in vivo Protein Interactions in All Bacterial Components by Förster Resonance Energy Transfer with the Superfolder mTurquoise2 ox-mNeongreen FRET Pair Authors:  Nils Y. Meiresonne, Elisa Consoli, Laureen M.Y. Mertens and Tanneke den Blaauwen, date: 12/05/2019, view: 51, Q&A: 0
This protocol was developed to qualitatively and quantitatively detect protein-protein interactions in all compartments of Escherichia coli by Förster Resonance Energy Transfer (FRET) using the Superfolder mTurquoise2 ox-mNeonGreen FRET pair (sfTq2ox-mNG). This FRET pair has more than twice the detection range for FRET ...
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Intracellular cAMP Measurements in Candida albicans Biofilms Authors:  Liuliu Jiang, Shengyan Chen, Kairui Sun, Peng Zhou and Xin Wei, date: 12/05/2019, view: 44, Q&A: 0
Candida albicans is the most common cause of fungal infections worldwide. Infection by C. albicans is closely associated with its ability to form a biofilm, closely packed communities of cells attached to the surfaces of human tissues and implanted devices, in or on the host. When tested for susceptibility to antifungals, such as ...
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Neuroscience

Intra-arterial Drug Delivery to the Ischemic Brain in Rat Middle Cerebral Artery Occlusion Model Authors:  Fereshteh Azedi, Masoud Mehrpour, Somaieh Kazemnejad, Kazem Mousavizadeh, Amir Hassan Zarnani and Mohammad Taghi Joghataei, date: 12/05/2019, view: 44, Q&A: 0
Rat transient middle cerebral artery occlusion (tMCAO) model is one of the most commonly used animal models in ischemic stroke studies. In the model, increasing safety and efficacy of therapeutic agent administration, such as stem cells and drugs directly to the ischemic brain using the internal carotid artery (ICA) is essential, because using the ...
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Trial-based Discrimination Procedure for Studying Drug Relapse in Rats Authors:  Veronica A Lennon, Megan B Brenner, Sophia J Weber, Lauren E Komer and Rajtarun Madangopal, date: 12/05/2019, view: 97, Q&A: 0
In abstinent drug addicts, cues formerly associated with drug-taking experiences gain relapse-inducing potency (‘incubate’) over time. Animal models of incubation may help in developing treatments for relapse prevention. However, these models have primarily focused on the role of conditioned stimuli (CSs) signaling drug delivery and not ...
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Mitochondrial Respiratory Measurements in Patient-derived Fibroblasts Authors:  Prashant Mishra, Ting Zhang, Ming Guo and David Chan, date: 12/05/2019, view: 43, Q&A: 0
Mitochondrial dysfunction is associated with a number of human diseases. As an example, we recently established in vivo Drosophila models of IBMPFD (Inclusion body myopathy, Paget disease, and frontotemporal dementia), and uncovered that human disease mutations of the p97/VCP (Valosin Containing Protein) gene behave as ...
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Evoked Potential Recordings of Auditory Brainstem Activity in the Mouse: An Optimized Method for the Assessment of Hearing Function of Mice Author:  Neil J. Ingham, date: 12/05/2019, view: 40, Q&A: 0
Hearing loss is a common sensory deficiency suffered by millions worldwide. It is a heterogeneous condition and genetics plays a critical role in its etiology. Gene variants can fundamentally alter hearing function, or predispose the auditory system towards loss of function resulting from other factors. In mouse studies of hearing loss and gene ...
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Plant Science

Localizing Genome Segments and Protein Products of a Multipartite Virus in Host Plant Cells Authors:  Marie-Stephanie Vernerey, Elodie Pirolles, Stephane Blanc and Anne Sicard, date: 12/05/2019, view: 61, Q&A: 0
A founding paradigm in virology is that the spatial unit of the viral replication cycle is an individual cell. This concept applied to multipartite viruses–which have a genome composed of two or more nucleic acid segments, each individually encapsulated–implies that all segments constituting a viral genome need to coinfect the same host cell for ...
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Systems Biology

Mapping the Mechanome–A Protocol for Simultaneous Live Imaging and Quantitative Analysis of Cell Mechanoadaptation and Ingression Authors:  Vina D. L. Putra, Iman Jalilian, Madeline Campbell, Kate Poole, Renee Whan, Florence Tomasetig and Melissa L. Knothe Tate, date: 12/05/2019, view: 50, Q&A: 0
Mechanomics, the mechanics equivalent of genomics, is a burgeoning field studying mechanical modulation of stem cell behavior and lineage commitment. Analogous to mechanical testing of a living material as it adapts and evolves, mapping of the mechanome necessitates the development of new protocols to assess changes in structure and function in ...
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