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https://www.readbyqxmd.com/read/28065310/analysis-of-postcytokinetic-roles-of-cytokinetic-components-in-caenorhabditis-elegans
#1
Y Chai, D Tian, W Li, G Ou
Cytokinesis requires the interplay between the cytoskeleton and plasma membrane. Emerging evidence indicates that some cytokinetic components are essential for the postcytokinetic events such as epithelium organization and neural development. We have recently developed live cell imaging and conditional knockout techniques to visualize cytokinetic proteins in Caenorhabditis elegans Q neuroblasts and separate their postcytokinetic functions from cytokinetic ones. Here we describe how the fluorescent reporter strains and conditional knockout C...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28063600/rna-in-situ-hybridization-characterization-of-non-enzymatic-derived-bovine-intervertebral-disc-cell-lineages-suggests-progenitor-cell-potential
#2
Petra Kraus, Rachel Yerden, Victoria Kocsis, Thomas Lufkin
Degeneration of the intervertebral disc (IVD) is a meritorious target for therapeutic cell based regenerative medicine approaches, however, controversy over what defines the precise identity of mature IVD cells and lack of single cell based quality control measures is of concern. Bos taurus and human IVDs are histologically more similar than is Mus musculus. The mature bovine IVD is well suited as model system for technology development to be translated into therapeutic cell based regenerative medicine applications...
January 4, 2017: Acta Histochemica
https://www.readbyqxmd.com/read/28060340/reprogramming-mouse-embryonic-fibroblasts-with-transcription-factors-to-induce-a-hemogenic-program
#3
Michael G Daniel, Carlos-Filipe Pereira, Jeffrey M Bernitz, Ihor R Lemischka, Kateri Moore
This protocol details the induction of a hemogenic program in mouse embryonic fibroblasts (MEFs) via overexpression of transcription factors (TFs). We first designed a reporter screen using MEFs from human CD34-tTA/TetO-H2BGFP (34/H2BGFP) double transgenic mice. CD34(+) cells from these mice label H2B histones with GFP, and cease labeling upon addition of doxycycline (DOX). MEFS were transduced with candidate TFs and then observed for the emergence of GFP(+) cells that would indicate the acquisition of a hematopoietic or endothelial cell fate...
December 16, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28060261/study-of-endoplasmic-reticulum-and-mitochondria-interactions-by-in-situ-proximity-ligation-assay-in-fixed-cells
#4
Emily Tubbs, Jennifer Rieusset
Structural interactions between the endoplasmic reticular (ER) and mitochondrial membranes, in domains known as mitochondria-associated membranes (MAM), are crucial hubs for cellular signaling and cell fate. Particularly, these inter-organelle contact sites allow the transfer of calcium from the ER to mitochondria through the voltage-dependent anion channel (VDAC)/glucose-regulated protein 75 (GRP75)/inositol 1,4,5-triphosphate receptor (IP3R) calcium channeling complex. While this subcellular compartment is under intense investigation in both physiological and pathological conditions, no simple and sensitive method exists to quantify the endogenous amount of ER-mitochondria contact in cells...
December 10, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28013130/biosynthesis-of-uniformly-labeled-13-c-and-14-c-arachidonic-acid-in-mortierella-alpina
#5
Jin V Lee, Ran Furman, Paul H Axelsen
Arachidonic acid (ARA) is one of the most abundant polyunsaturated fatty acids (PUFAs) in the mammalian brain. Many enzymatically- and nonenzymatically-produced metabolic products have important and potent pharmacological properties. However, uniformly isotope labeled forms of ARA are not commercially available for studying the metabolic fates of ARA. This study describes a simple and efficient protocol for the biosynthesis of U-(13)C-ARA from U-(13)C-glucose, and U-(14)C-ARA from U-(14)C-glucose by Mortierella alpina...
March 2017: Bioresource Technology
https://www.readbyqxmd.com/read/27965839/cognitive-behavioural-therapy-for-psychopathology-in-relatives-of-missing-persons-study-protocol-for-a-pilot-randomised-controlled-trial
#6
Lonneke I M Lenferink, Ineke Wessel, Jos de Keijser, Paul A Boelen
BACKGROUND: It is hypothesized that the grieving process of relatives of missing persons is complicated by having to deal with uncertainty about the fate of their loved one. We developed a cognitive behavioural therapy (CBT) with mindfulness that focuses on dealing with this uncertainty. In this article, we elucidate the rationale of a pilot randomised controlled trial (RCT) for testing the feasibility and potential effectiveness of this CBT for reducing symptoms of psychopathology in relatives of missing persons...
2016: Pilot and Feasibility Studies
https://www.readbyqxmd.com/read/27956115/future-perspectives-in-adult-stem-cell-turnover-implications-for-endocrine-physiology-and-disease
#7
Clara V Alvarez, Fernando Oroz-Gonjar, Montserrat Garcia-Lavandeira
No abstract text is available yet for this article.
December 9, 2016: Molecular and Cellular Endocrinology
https://www.readbyqxmd.com/read/27924563/high-throughput-in-vitro-identification-of-direct-mapk-erk-substrates
#8
Rona Grossman, Ze'ev Paroush
Phosphorylation mediated by cellular protein kinases is an effective mechanism employed by an organism to regulate central processes such as cell-cycle progression, metabolic pathways, cytoskeletal function, cell migration and differentiation. Thus, for example, various signaling pathways utilize sequential phosphorylation events to relay external cues from the cell surface to the nucleus, where eventually gene expression profiles are altered and, consequently, changes in cell fates and function are induced...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27906170/generating-high-purity-cardiac-and-endothelial-derivatives-from-patterned-mesoderm-using-human-pluripotent-stem-cells
#9
Nathan J Palpant, Lil Pabon, Clayton E Friedman, Meredith Roberts, Brandon Hadland, Rebecca J Zaunbrecher, Irwin Bernstein, Ying Zheng, Charles E Murry
Human pluripotent stem cells (hPSCs) provide a valuable model for the study of human development and a means to generate a scalable source of cells for therapeutic applications. This protocol specifies cell fate efficiently into cardiac and endothelial lineages from hPSCs. The protocol takes 2 weeks to complete and requires experience in hPSC culture and differentiation techniques. Building on lessons taken from early development, this monolayer-directed differentiation protocol uses different concentrations of activin A and bone morphogenetic protein 4 (BMP4) to polarize cells into mesodermal subtypes that reflect mid-primitive-streak cardiogenic mesoderm and posterior-primitive-streak hemogenic mesoderm...
January 2017: Nature Protocols
https://www.readbyqxmd.com/read/27830548/fetal-liver-stem-progenitor-cell-transplantation-a-model-to-study-tissue-mass-replacement-and-cell-based-therapies
#10
Mladen I Yovchev, Michael Oertel
Liver transplantation is the only therapeutic treatment for patients with end-stage liver diseases. However, donor organ scarcity is the major limitation, and therefore, alternative strategies are urgently needed. The ultimate goal for successful cell-based therapies is the ability of transplanted cells to efficiently engraft and reconstitute injured liver mass. To evaluate the repopulation capacity of transplanted cells, it is essential to identify their specific characteristics, as well as to study the mechanism(s) Through which transplanted donor cells replace tissue mass in hepatic microenvironments, using well-established cell transplantation models...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27820711/when-protocols-become-fairy-tales-and-gods-remain-buried-under-excerpts-from-the-diary-of-forensic-experts-at-ground-zero-during-the-mega-quake-that-hit-nepal
#11
Jenash Acharya, Rijen Shrestha, Pramod Kumar Shrestha, Tanuj Kanchan, Kewal Krishan
More than a year has passed since the mega quake hit Nepal. Although a lot is written about the lacunae in disaster preparedness, lessons learnt, and public health concerns after the aftermath, nothing much has been written about the fate of the deceased. Although saving takes priority after a disaster, the management of dead bodies also requires immediate attention because the process of identification becomes more difficult and expensive as time passes. This article shares the firsthand experiences of forensic experts at ground zero during the quake that may be useful to forensic experts responsible for handling such situations worldwide...
November 4, 2016: American Journal of Forensic Medicine and Pathology
https://www.readbyqxmd.com/read/27815897/synchronization-and-desynchronization-of-cells-by-interventions-on-the-spindle-assembly-checkpoint
#12
Mohamed Jemaà, Gwenola Manic, Ilio Vitale
Cell cycle checkpoints are surveillance mechanisms that sequentially and continuously monitor cell cycle progression thereby contributing to the preservation of genetic stability. Among them, the spindle assembly checkpoint (SAC) prevents the occurrence of abnormal divisions by halting the metaphase to anaphase transition following the detection of erroneous microtubules-kinetochore attachment(s). Most synchronization strategies are based on the activation of cell cycle checkpoints to enrich the population of cells in a specific phase of the cell cycle...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27807896/transient-cnp-expression-by-early-progenitors-causes-cre-lox-based-reporter-lines-to-map-profoundly-different-fates
#13
Reshmi Tognatta, Wenjing Sun, Sandra Goebbels, Klaus-Armin Nave, Akiko Nishiyama, Susanne Schoch, Leda Dimou, Dirk Dietrich
NG2 expressing oligodendroglial precursor cells are ubiquitous in the central nervous system and the only cell type cycling throughout life. Previous fate mapping studies have remained inconsistent regarding the question whether NG2 cells are capable of generating certain types of neurons. Here, we use CNP-Cre mice to map the fate of a sub-population of NG2 cells assumed to be close to differentiation. When crossing these mice with the ROSA26/YFP Cre-reporter line we discovered large numbers of reporter-expressing pyramidal neurons in the piriform and dorsal cortex...
February 2017: Glia
https://www.readbyqxmd.com/read/27805614/detection-of-the-ph-dependent-activity-of-escherichia-coli-chaperone-hdeb-in-vitro-and-in-vivo
#14
Jan-Ulrik Dahl, Philipp Koldewey, James C A Bardwell, Ursula Jakob
Bacteria are frequently exposed to environmental changes, such as alterations in pH, temperature, redox status, light exposure or mechanical force. Many of these conditions cause protein unfolding in the cell and have detrimental impact on the survival of the organism. A group of unrelated, stress-specific molecular chaperones have been shown to play essential roles in the survival of these stress conditions. While fully folded and chaperone-inactive before stress, these proteins rapidly unfold and become chaperone-active under specific stress conditions...
October 23, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27796959/techniques-for-the-reprogramming-of-exogenous-stem-progenitor-cell-populations-towards-a-mammary-epithelial-cell-fate
#15
Gilbert H Smith, Corinne A Boulanger
This chapter considers the techniques necessary and required for the reprogramming of exogenous stem/progenitor cell populations towards a mammary epithelial cell fate. The protocols describe how to isolate cells from alternate mouse organs such as testicles of male mice and mix them with mammary cells to generate chimeric glands comprised of male and female epithelial cells that are fully competent. During the reformation of mammary stem cell niches by dispersed epithelial cells, in the context of the intact epithelium-free mammary stroma, non-mammary cells are sequestered and reprogrammed to perform mammary epithelial cell functions including those ascribed to mammary stem/progenitor cells...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27782412/communication-thermal-unimolecular-decomposition-of-syn-ch3choo-a-kinetic-study
#16
Thanh Lam Nguyen, Laura McCaslin, Michael C McCarthy, John F Stanton
The thermal decomposition of syn-ethanal-oxide (syn-CH3CHOO) through vinyl hydrogen peroxide (VHP) leading to hydroxyl radical is characterized using a modification of the HEAT thermochemical protocol. The isomerization step of syn-CH3CHOO to VHP via a 1,4 H-shift, which involves a moderate barrier of 72 kJ/mol, is found to be rate determining. A two-dimensional master equation approach, in combination with semi-classical transition state theory, is employed to calculate the time evolution of various species as well as to obtain phenomenological rate coefficients...
October 7, 2016: Journal of Chemical Physics
https://www.readbyqxmd.com/read/27744003/neural-organoids-for-disease-phenotyping-drug-screening-and-developmental-biology-studies
#17
Brigham J Hartley, Kristen J Brennand
Human induced pluripotent stem cells (hiPSCs) can theoretically yield limitless supplies of cells fated to any cell type that comprise the human organism, making them a new tool by which to potentially overcome caveats in current biomedical research. In vitro derivation of central nervous system (CNS) cell types has the potential to provide material for drug discovery and validation, safety and toxicity assays, cell replacement therapy and the elucidation of previously unknown disease mechanisms. However, current two-dimensional (2D) CNS differentiation protocols do not faithfully recapitulate the spatial organization of heterogeneous tissue, nor the cell-cell interactions, cell-extracellular matrix interactions, or specific physiological functions generated within complex tissue such as the brain...
October 12, 2016: Neurochemistry International
https://www.readbyqxmd.com/read/27737438/assessment-of-scaffolding-properties-for-chondrogenic-differentiation-of-adipose-derived-mesenchymal-stem-cells-in-nasal-reconstruction
#18
Serban San-Marina, Ayushman Sharma, Stephen G Voss, Jeffrey R Janus, Grant S Hamilton
Importance: Nasal reconstruction in patients who are missing a significant amount of structural nasal support remains a difficult challenge. One challenge is the deficiency of cartilage left within the nose as a consequence of rhinectomy or a midline destructive disease. Historically, the standard donor source for large quantities of native cartilage has been costal cartilage. Objective: To enable the development of protocols for new mesenchymal stem cell technologies as alternative procedures with reduced donor site morbidity, risk of infection and extrusion...
October 13, 2016: JAMA Facial Plastic Surgery
https://www.readbyqxmd.com/read/27735934/self-organization-of-human-embryonic-stem-cells-on-micropatterns
#19
Alessia Deglincerti, Fred Etoc, M Cecilia Guerra, Iain Martyn, Jakob Metzger, Albert Ruzo, Mijo Simunovic, Anna Yoney, Ali H Brivanlou, Eric Siggia, Aryeh Warmflash
Fate allocation in the gastrulating embryo is spatially organized as cells differentiate into specialized cell types depending on their positions with respect to the body axes. There is a need for in vitro protocols that allow the study of spatial organization associated with this developmental transition. Although embryoid bodies and organoids can exhibit some spatial organization of differentiated cells, methods that generate embryoid bodies or organoids do not yield consistent and fully reproducible results...
November 2016: Nature Protocols
https://www.readbyqxmd.com/read/27734355/transplantation-as-a-quantitative-assay-to-study-mammalian-male-germline-stem-cells
#20
Aileen R Helsel, Jon M Oatley
In mammals, the activities of spermatogonial stem cells (SSCs) provide the foundation for continual spermatogenesis throughout a male's reproductive lifetime. At present, the defining characteristics of SSCs and mechanisms controlling their fate decisions are not well understood. Transplantation is a definitive functional measure of stem cell capacity for male germ cells that can be used as an assay to provide an unequivocal quantification of the SSC content in an experimental cell population. Here, we discuss the procedure for mice and provide protocols for preparing donor germ cell suspensions from testes directly or primary cultures of spermatogonia for transplantation, enriching for SSCs, preparing recipient males, microinjection into recipient testes, and considerations for experimental design...
2017: Methods in Molecular Biology
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