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FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts

p53 over expression in yeast results in cell death with typical markers of apoptosis such as DNA fragmentation and phosphatidylserine externalization. We aimed to substitute/supplement classical fluorescent techniques (TUNEL, Annexin V, ROS detection) usually used to detect biochemical changes occur...

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Autores principales: Mihoubi, Wafa, Sahli, Emna, Gargouri, Ali, Amiel, Caroline
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5507515/
https://www.ncbi.nlm.nih.gov/pubmed/28704406
http://dx.doi.org/10.1371/journal.pone.0180680
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author Mihoubi, Wafa
Sahli, Emna
Gargouri, Ali
Amiel, Caroline
author_facet Mihoubi, Wafa
Sahli, Emna
Gargouri, Ali
Amiel, Caroline
author_sort Mihoubi, Wafa
collection PubMed
description p53 over expression in yeast results in cell death with typical markers of apoptosis such as DNA fragmentation and phosphatidylserine externalization. We aimed to substitute/supplement classical fluorescent techniques (TUNEL, Annexin V, ROS detection) usually used to detect biochemical changes occurring during yeast apoptosis mediated by p53 over expression and the effect of anti-apoptotic purified molecules from Nigel (Nigella sativa) extracts on these same yeasts by the label free technique of FTIR spectroscopy. The comparison of the entire IR spectra highlighted clear modifications between apoptotic p53-expressing yeasts and normal ones. More precisely, DNA damage was detected by the decrease of band intensities at 1079 and 1048 cm(-1). While phosphatidylserine exposure was followed by the increase of νsCH(2) and νasCH(2) bands of unsaturated fatty acids that were exhibited at 2855 and 2926 cm(-1), and the appearance of the C = O ester functional group band at 1740 cm(-1). In a second step, this FTIR approach was used to estimate the effect of a purified fraction of the Nigel extract. The modulation of band intensities specific to DNA and membrane status was in agreement with apoptosis supression in presence of the Nigel extracts. FTIR spectroscopy is thus proven to be a very reliable technique to monitor the apoptotic cell death in yeast and to be used as a means of evaluating the biomolecules effect on yeast survival.
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spelling pubmed-55075152017-07-25 FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts Mihoubi, Wafa Sahli, Emna Gargouri, Ali Amiel, Caroline PLoS One Research Article p53 over expression in yeast results in cell death with typical markers of apoptosis such as DNA fragmentation and phosphatidylserine externalization. We aimed to substitute/supplement classical fluorescent techniques (TUNEL, Annexin V, ROS detection) usually used to detect biochemical changes occurring during yeast apoptosis mediated by p53 over expression and the effect of anti-apoptotic purified molecules from Nigel (Nigella sativa) extracts on these same yeasts by the label free technique of FTIR spectroscopy. The comparison of the entire IR spectra highlighted clear modifications between apoptotic p53-expressing yeasts and normal ones. More precisely, DNA damage was detected by the decrease of band intensities at 1079 and 1048 cm(-1). While phosphatidylserine exposure was followed by the increase of νsCH(2) and νasCH(2) bands of unsaturated fatty acids that were exhibited at 2855 and 2926 cm(-1), and the appearance of the C = O ester functional group band at 1740 cm(-1). In a second step, this FTIR approach was used to estimate the effect of a purified fraction of the Nigel extract. The modulation of band intensities specific to DNA and membrane status was in agreement with apoptosis supression in presence of the Nigel extracts. FTIR spectroscopy is thus proven to be a very reliable technique to monitor the apoptotic cell death in yeast and to be used as a means of evaluating the biomolecules effect on yeast survival. Public Library of Science 2017-07-12 /pmc/articles/PMC5507515/ /pubmed/28704406 http://dx.doi.org/10.1371/journal.pone.0180680 Text en © 2017 Mihoubi et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Mihoubi, Wafa
Sahli, Emna
Gargouri, Ali
Amiel, Caroline
FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts
title FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts
title_full FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts
title_fullStr FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts
title_full_unstemmed FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts
title_short FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts
title_sort ftir spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by nigella sativa extracts
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5507515/
https://www.ncbi.nlm.nih.gov/pubmed/28704406
http://dx.doi.org/10.1371/journal.pone.0180680
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