scholarly journals Usefulness of a Non-invasive Reporter System for Monitoring Reprogramming State in Pig Cells: Results of a Cell Fusion Experiment

2010 ◽  
Vol 56 (4) ◽  
pp. 363-369 ◽  
Author(s):  
Akio OZAWA ◽  
Eri AKASAKA ◽  
Satoshi WATANABE ◽  
Mitsutoshi YOSHIDA ◽  
Kazuchika MIYOSHI ◽  
...  
Keyword(s):  
Cell Fusion ◽  
Reporter System ◽  
Non Invasive ◽  
A Cell

Quantitative cell fusion: the fusion sensitivity (FS) potential

1981 ◽  
Vol 49 (1) ◽  
pp. 87-97
Author(s):  
D. Rohme
Keyword(s):  
Cell Line ◽  
Cell Fusion ◽  
Cell Lines ◽  
Dose Response ◽  
Sendai Virus ◽  
Biological Significance ◽  
Diploid Cell ◽  
Mammalian Cell Lines ◽  
Virus Dose ◽  
A Cell

The dose response of Sendai virus-induced cell fusion was studied in 10 mammalian cell lines, comprising 5 continuous and 5 diploid cell lines originating from 5 species. The extent of fusion was calculated using a parameter directly proportional to the number of fusion events (t-parameter). At lower levels of fusion the dose response was found to be based on the same simple kinetic rules in all cell lines and was defined by the formula: t = FS. FAU/(I + FS. FAU), where FS (fusion sensitivity) is a cell-specific constant of the fusion rate and FAU (fusion activity units) is the virus dose. The FS potential of a cell line was determined as the linear regression coefficient of the fusion index (t/(I - t)) on the virus dose. At higher levels of fusion, when the fusion extent reached cell-line-specific maximal levels, the dose response was not as uniform. In general, and particularly in the cases of the diploid cell lines, these maximal levels were directly proportional to the FS potentials. Thus, it was concluded that the FS potential is the basic quantitative feature, which expresses the cellular fusion efficiency. The fact that FS varied extensively between cell lines, but at the same time apparently followed certain patterns (being higher in continuous compared to diploid cell lines and being related to the species of origin of the cells), emphasizes it biological significance as well as its possible usefulness in studies of the efficiency of various molecular interactions in the cell membrane/cytoskeleton system.

scholarly journals Detection of Amyotrophic Lateral Sclerosis (ALS) via Acoustic Analysis

2018 ◽  
Author(s):  
Raquel Norel ◽  
Mary Pietrowicz ◽  
Carla Agurto ◽  
Shay Rishoni ◽  
Guillermo Cecchi
Keyword(s):  
Disease Progression ◽  
Acoustic Analysis ◽  
Voice Quality ◽  
Non Invasive ◽  
Frequent Monitoring ◽  
In The Wild ◽  
A Cell ◽  
Speech Features ◽  
Severity Of The Disease ◽  
Lateral Sclerosis

ABSTRACTALS is a fatal neurodegenerative disease with no cure. Experts typically measure disease progression via the ALSFRS-R score, which includes measurements of various abilities known to decline. We propose instead the use of speech analysis as a proxy for ALS progression. This technique enables 1) frequent non-invasive, inexpensive, longitudinal analysis, 2) analysis of data recorded in the wild, and 3) creation of an extensive ALS databank for future analysis. Patients and trained medical professionals need not be co-located, enabling more frequent monitoring of more patients from the convenience of their own homes. The goals of this study are the identification of acoustic speech features in naturalistic contexts which characterize disease progression and development of machine models which can recognize the presence and severity of the disease. We evaluated subjects from the Prize4Life Israel dataset, using a variety of frequency, spectral, and voice quality features. The dataset was generated using the ALS Mobile Analyzer, a cell-phone app that collects data regarding disease progress using a self-reported ALSFRS-R questionnaire and several active tasks that measure speech and motor skills. Classification via leave-five-subjects-out cross-validation resulted in an accuracy rate of 79% (61% chance) for males and 83% (52% chance) for females.

scholarly journals A Cell-Based High-Throughput Assay for the Screening of Small-Molecule Inhibitors of p53–MDM2 Interaction

2011 ◽  
Vol 16 (4) ◽  
pp. 450-456 ◽  
Author(s):  
Jing Li ◽  
Shuyong Zhang ◽  
Linghuan Gao ◽  
Ying Chen ◽  
Xin Xie
Keyword(s):  
High Throughput ◽  
Luciferase Reporter ◽  
Reporter System ◽  
Direct Inhibition ◽  
A Cell ◽  
Luciferase Reporter System ◽  
Novel Strategy ◽  
High Throughput Assay ◽  
Two Hybrid ◽  
Rule Out

The p53 tumor suppressor is a potent transcription factor that regulates cell growth inhibition and apoptosis. The oncoprotein MDM2 suppresses p53 activity by direct inhibition of its transcriptional activity and enhances the degradation of p53 via the ubiquitin–proteosome pathway. Overexpression of MDM2, found in many human tumors, impairs p53-mediated cell death effectively. Inhibition of the p53–MDM2 interaction can stabilize p53 and may offer a novel strategy for cancer therapy. To search for new inhibitors of the p53–MDM2 interaction, the authors developed a cell-based high-throughput assay system based on mammalian two-hybrid technology. They also used a dual-luciferase reporter system to rule out false- positive hits due to the cytotoxic effect of compounds. Using this assay, they screened a library consisting of 3840 compounds and identified one compound that activates p53 pathway and induces growth arrest in tumor cells.

scholarly journals The Anticoagulant Nafamostat Potently Inhibits SARS-CoV-2 S Protein-Mediated Fusion in a Cell Fusion Assay System and Viral Infection In Vitro in a Cell-Type-Dependent Manner

Viruses ◽  
2020 ◽  
Vol 12 (6) ◽  
pp. 629 ◽  
Author(s):  
Mizuki Yamamoto ◽  
Maki Kiso ◽  
Yuko Sakai-Tagawa ◽  
Kiyoko Iwatsuki-Horimoto ◽  
Masaki Imai ◽  
...  
Keyword(s):  
Cell Fusion ◽  
Assay System ◽  
Dependent Manner ◽  
Cell Type ◽  
S Protein ◽  
Candidate Drug ◽  
A Cell ◽  
Transmembrane Serine Protease ◽  
Nafamostat Mesylate

Although infection by SARS-CoV-2, the causative agent of coronavirus pneumonia disease (COVID-19), is spreading rapidly worldwide, no drug has been shown to be sufficiently effective for treating COVID-19. We previously found that nafamostat mesylate, an existing drug used for disseminated intravascular coagulation (DIC), effectively blocked Middle East respiratory syndrome coronavirus (MERS-CoV) S protein-mediated cell fusion by targeting transmembrane serine protease 2 (TMPRSS2), and inhibited MERS-CoV infection of human lung epithelium-derived Calu-3 cells. Here we established a quantitative fusion assay dependent on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) S protein, angiotensin I converting enzyme 2 (ACE2) and TMPRSS2, and found that nafamostat mesylate potently inhibited the fusion while camostat mesylate was about 10-fold less active. Furthermore, nafamostat mesylate blocked SARS-CoV-2 infection of Calu-3 cells with an effective concentration (EC)50 around 10 nM, which is below its average blood concentration after intravenous administration through continuous infusion. On the other hand, a significantly higher dose (EC50 around 30 μM) was required for VeroE6/TMPRSS2 cells, where the TMPRSS2-independent but cathepsin-dependent endosomal infection pathway likely predominates. Together, our study shows that nafamostat mesylate potently inhibits SARS-CoV-2 S protein-mediated fusion in a cell fusion assay system and also inhibits SARS-CoV-2 infection in vitro in a cell-type-dependent manner. These findings, together with accumulated clinical data regarding nafamostat’s safety, make it a likely candidate drug to treat COVID-19.

scholarly journals Targeted disruption of genes for gp138, a cell-fusion-related protein in Dictyostelium discoideum, revealed the existence of a third gene

1996 ◽  
Vol 38 (3) ◽  
pp. 271-279 ◽  
Author(s):  
Nobuyuki Yamaguchi ◽  
Mikihiko Higa ◽  
Kazuhiro Aiba ◽  
Hui Fang ◽  
Yoshimasa Tanaka ◽  
...  
Keyword(s):  
Dictyostelium Discoideum ◽  
Cell Fusion ◽  
Related Protein ◽  
Targeted Disruption ◽  
A Cell

scholarly journals Human cord blood CD34+ progenitor cells acquire functional cardiac properties through a cell fusion process

2011 ◽  
Vol 300 (5) ◽  
pp. H1875-H1884 ◽  
Author(s):  
Daniele Avitabile ◽  
Alessia Crespi ◽  
Chiara Brioschi ◽  
Valeria Parente ◽  
Gabriele Toietta ◽  
...  
Keyword(s):  
Cord Blood ◽  
Cell Fusion ◽  
Fluorescent Protein ◽  
Lentiviral Vector ◽  
Fusion Process ◽  
Resting Potential ◽  
Human Cord Blood ◽  
Neonatal Cardiomyocytes ◽  
Cord Blood Cd34 ◽  
A Cell

The efficacy of cardiac repair by stem cell administration relies on a successful functional integration of injected cells into the host myocardium. Safety concerns have been raised about the possibility that stem cells may induce foci of arrhythmia in the ischemic myocardium. In a previous work ( 36 ), we showed that human cord blood CD34+ cells, when cocultured on neonatal mouse cardiomyocytes, exhibit excitation-contraction coupling features similar to those of cardiomyocytes, even though no human genes were upregulated. The aims of the present work are to investigate whether human CD34+ cells, isolated after 1 wk of coculture with neonatal ventricular myocytes, possess molecular and functional properties of cardiomyocytes and to discriminate, using a reporter gene system, whether cardiac differentiation derives from a (trans)differentiation or a cell fusion process. Umbilical cord blood CD34+ cells were isolated by a magnetic cell sorting method, transduced with a lentiviral vector carrying the enhanced green fluorescent protein (EGFP) gene, and seeded onto primary cultures of spontaneously beating rat neonatal cardiomyocytes. Cocultured EGFP+/CD34+-derived cells were analyzed for their electrophysiological features at different time points. After 1 wk in coculture, EGFP+ cells, in contact with cardiomyocytes, were spontaneously contracting and had a maximum diastolic potential (MDP) of −53.1 mV, while those that remained isolated from the surrounding myocytes did not contract and had a depolarized resting potential of −11.4 mV. Cells were then resuspended and cultured at low density to identify EGFP+ progenitor cell derivatives. Under these conditions, we observed single EGFP+ beating cells that had acquired an hyperpolarization-activated current typical of neonatal cardiomyocytes (EGFP+ cells, −2.24 ± 0.89 pA/pF; myocytes, −1.99 ± 0.63 pA/pF, at −125 mV). To discriminate between cell autonomous differentiation and fusion, EGFP+/CD34+ cells were cocultured with cardiac myocytes infected with a red fluorescence protein-lentiviral vector; under these conditions we found that 100% of EGFP+ cells were also red fluorescent protein positive, suggesting cell fusion as the mechanism by which cardiac functional features are acquired.

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