Mammalia

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    Protocols in Current Issue
    Real-time Three-dimensional Tracking of Endocytic Vesicles
    Authors:  Stephanie Duhamel and Kossay Zaoui, date: 10/20/2020, view: 203, Q&A: 0
    Endocytic trafficking and recycling are fundamental cellular processes that control essential functions such as signaling protein complexes transport and membrane identity. The small GTPase Rabs are indispensable component of the endosomal recycling ...
    Derivation of Induced Pluripotent Stem Cells from Human Fibroblasts Using a Non-integrative System in Feeder-free Conditions
    Authors:  Alvaro A. Beltran, Sarahi G. Molina and Adriana S. Beltran, date: 10/20/2020, view: 136, Q&A: 0
    Induced pluripotent stem cells (iPSCs) are genetically reprogrammed somatic cells that exhibit features identical to those of embryonic stem cells (ESCs). Multiple approaches are available to derive iPSCs, among which the Sendai virus is the most ...
    Quantification of Salivary Charged Metabolites Using Capillary Electrophoresis Time-of-flight-mass Spectrometry
    Authors:  Masahiro Sugimoto, Sana Ota, Miku Kaneko, Ayame Enomoto and Tomoyoshi Soga, date: 10/20/2020, view: 112, Q&A: 0
    Salivary metabolomics have provided the potentials to detect both oral and systemic diseases. Capillary electrophoresis time-of-flight-mass spectrometry (CE-TOFMS) enables the identification and quantification of various charged metabolites. This ...
    Staining and Quantitative Analysis of Myelinating Oligodendrocytes in the Mouse Grey Matter
    Authors:  Matthew Swire and Charles ffrench-Constant, date: 10/20/2020, view: 108, Q&A: 0
    Oligodendrocytes generate distinct patterns of myelination throughout the CNS. Variations in myelination along axons may enable neurons to fine-tune conduction velocities and alter signal synchronisation. Here we outline a staining protocol ...
    Analyzing (Re)Capping of mRNA Using Transcript Specific 5' End Sequencing
    Authors:  Daniel del Valle Morales and Daniel R. Schoenberg, date: 10/20/2020, view: 170, Q&A: 0
    The 5′ cap is a ubiquitous feature of eukaryotic mRNAs. It is added in the nucleus onto newly synthesized pre-mRNA, and in the cytoplasm onto mRNAs after decapping or endonuclease cleavage. Cytoplasmic recapping can occur after loss of the cap at ...
    Estimation of the Minimum Number of Replication Origins Per Chromosome in any Organism
    Author:  Marcelo S. da Silva, date: 10/20/2020, view: 173, Q&A: 0
    Eukaryote nuclear genomes predominantly replicate through multiple replication origins. The number of replication origins activated per chromosome during the S-phase duration may vary according to many factors, but the predominant one is replication ...
    Real-time Three-dimensional Tracking of Endocytic Vesicles
    Authors:  Stephanie Duhamel and Kossay Zaoui, date: 10/20/2020, view: 203, Q&A: 0
    [Abstract] Endocytic trafficking and recycling are fundamental cellular processes that control essential functions such as signaling protein complexes transport and membrane identity. The small GTPase Rabs are indispensable component of the endosomal recycling machinery. The Rabs bind to effectors to mediate their functions, such as protein sorting and ...
    Derivation of Induced Pluripotent Stem Cells from Human Fibroblasts Using a Non-integrative System in Feeder-free Conditions
    Authors:  Alvaro A. Beltran, Sarahi G. Molina and Adriana S. Beltran, date: 10/20/2020, view: 136, Q&A: 0
    [Abstract] Induced pluripotent stem cells (iPSCs) are genetically reprogrammed somatic cells that exhibit features identical to those of embryonic stem cells (ESCs). Multiple approaches are available to derive iPSCs, among which the Sendai virus is the most effective at reprogramming different cell types. Here we describe a rapid, efficient, safe, and ...
    Quantification of Salivary Charged Metabolites Using Capillary Electrophoresis Time-of-flight-mass Spectrometry
    Authors:  Masahiro Sugimoto, Sana Ota, Miku Kaneko, Ayame Enomoto and Tomoyoshi Soga, date: 10/20/2020, view: 112, Q&A: 0
    [Abstract] Salivary metabolomics have provided the potentials to detect both oral and systemic diseases. Capillary electrophoresis time-of-flight-mass spectrometry (CE-TOFMS) enables the identification and quantification of various charged metabolites. This method has been employed to biomarker discoveries using human saliva samples, especially for various ...
    Staining and Quantitative Analysis of Myelinating Oligodendrocytes in the Mouse Grey Matter
    Authors:  Matthew Swire and Charles ffrench-Constant, date: 10/20/2020, view: 108, Q&A: 0
    [Abstract] Oligodendrocytes generate distinct patterns of myelination throughout the CNS. Variations in myelination along axons may enable neurons to fine-tune conduction velocities and alter signal synchronisation. Here we outline a staining protocol permitting the assessment of the number and length of myelin sheaths formed by oligodendrocyte in the mouse ...
    Analyzing (Re)Capping of mRNA Using Transcript Specific 5' End Sequencing
    Authors:  Daniel del Valle Morales and Daniel R. Schoenberg, date: 10/20/2020, view: 170, Q&A: 0
    [Abstract] The 5′ cap is a ubiquitous feature of eukaryotic mRNAs. It is added in the nucleus onto newly synthesized pre-mRNA, and in the cytoplasm onto mRNAs after decapping or endonuclease cleavage. Cytoplasmic recapping can occur after loss of the cap at the native 5′ end, or downstream within the body of the mRNA. The identification and location of ...
    Estimation of the Minimum Number of Replication Origins Per Chromosome in any Organism
    Author:  Marcelo S. da Silva, date: 10/20/2020, view: 173, Q&A: 0
    [Abstract] Eukaryote nuclear genomes predominantly replicate through multiple replication origins. The number of replication origins activated per chromosome during the S-phase duration may vary according to many factors, but the predominant one is replication stress. Several studies have applied different approaches to estimate the number and map the ...
    TetR Regulated in vivo Repression Technology to Identify Conditional Gene Silencing in Genetically Engineerable Bacteria Using Vibrio cholerae Murine Infections as Model System
    Authors:  Franz G. Zingl, Fabian Mitterer, Himadri B. Thapa and Stefan Schild, date: 10/05/2020, view: 199, Q&A: 0
    [Abstract] Investigation of bacterial gene regulation upon environmental changes is still a challenging task. For example, Vibrio cholerae, a pathogen of the human gastrointestinal tract, faces diverse transient conditions in different compartments upon oral ingestion. Genetic reporter systems have been demonstrated to be extremely powerful tools to ...
    Using RNA Sequencing and Spike-in RNAs to Measure Intracellular Abundance of lncRNAs and mRNAs
    Authors:  Megan D. Schertzer, McKenzie M. Murvin and J. Mauro Calabrese, date: 10/05/2020, view: 225, Q&A: 0
    [Abstract] Long noncoding RNAs (lncRNAs) play essential roles in normal physiology and in disease but their mechanisms of action can be challenging to identify. For mechanistic studies, it is often useful to know a lncRNA’s intracellular abundance, i.e., approximately how many molecules of the lncRNA are present in a typical cell of a cell-type of ...
    An Operant Conditioning Model Combined with a Chemogenetic Approach to Study the Neurobiology of Food Addiction in Mice
    Authors:  Elena Martín-García, Laura Domingo-Rodriguez and Rafael Maldonado, date: 10/05/2020, view: 265, Q&A: 0
    [Abstract] The study of food addiction comprises 3 hallmarks that include the persistence to response without an outcome, the strong motivation for palatable food, and the loss of inhibitory control over food intake that leads to compulsive behavior in addicted individuals. The complex multifactorial nature of this disorder and the unknown neurobiological ...
    Radioactive Assay of in vitro Glutamylation Activity of the Legionella pneumophila Effector Protein SidJ
    Authors:  Alan G. Sulpizio, Jung-Ho Shin, Marena E. Minelli and Yuxin Mao, date: 10/05/2020, view: 278, Q&A: 0
    [Abstract] The Legionella effector protein SidJ has recently been identified to perform polyglutamylation on another Legionella effector, SdeA, ablating SdeA’s activity. SidJ is a kinase-like protein that requires the small eukaryotic protein calmodulin to perform glutamylation. Glutamylation is a relatively uncommon type of ...



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