Deepening of lipidome annotation by associating cross-metathesis reaction with mass spectrometry: application to an in vitro model of corneal toxicity

2021 ◽  
Author(s):  
Romain Magny ◽  
Anne Regazzetti ◽  
Karima Kessal ◽  
Christophe Baudouin ◽  
Stéphane Mélik-Parsadaniantz ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
In Vitro Model ◽  
Metathesis Reaction ◽  
Cross Metathesis ◽  
Corneal Toxicity ◽  
Vitro Model ◽  
Mass Spectrometry Application

scholarly journals In-Depth Lipidome Annotation Through an Operatively Simple Method Combining Cross-Metathesis Reaction and Tandem Mass Spectrometry

2021 ◽  
Author(s):  
Romain MAGNY ◽  
Anne Regazzetti ◽  
Karima Kessal ◽  
Christophe Baudouin ◽  
Stéphane Mélik-Parsadaniantz ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Tandem Mass ◽  
Metathesis Reaction ◽  
Simple Method ◽  
Cross Metathesis ◽  
Corneal Toxicity ◽  
Vitro Model ◽  
Structure Annotation

The in-depth knowledge of lipid biological functions calls for a comprehensive lipid structure annotation that implies implementing a method to locate fatty acids unsaturations. To address this challenge we have associated Grubbs' cross metathesis reaction and liquid chromatography hyphenated to tandem mass spectrometry. The pretreatment of lipids containing samples by Grubbs' catalyst and an appropriate alken generates substituted lipids through cross-metathesis reaction under mild, chemoselective and highly reproducible conditions. A systematic LC-MS/MS analysis of the reaction mixture allows locating unambigouslt the double bounds in fatty acid side chains. This method has en successfully applied at a nanomole scale to commerical standard mixtures as well as in lipid extracts from an in vitro model of corneal toxicity.<br><div><a></a><a></a><a></a><a></a></div><div><a></a><a></a><a></a> </div>

scholarly journals In-Depth Lipidome Annotation Through an Operatively Simple Method Combining Cross-Metathesis Reaction and Tandem Mass Spectrometry

2021 ◽  
Author(s):  
Romain MAGNY ◽  
Anne Regazzetti ◽  
Karima Kessal ◽  
Christophe Baudouin ◽  
Stéphane Mélik-Parsadaniantz ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Tandem Mass ◽  
Metathesis Reaction ◽  
Simple Method ◽  
Cross Metathesis ◽  
Corneal Toxicity ◽  
Vitro Model ◽  
Structure Annotation

The in-depth knowledge of lipid biological functions calls for a comprehensive lipid structure annotation that implies implementing a method to locate fatty acids unsaturations. To address this challenge we have associated Grubbs' cross metathesis reaction and liquid chromatography hyphenated to tandem mass spectrometry. The pretreatment of lipids containing samples by Grubbs' catalyst and an appropriate alken generates substituted lipids through cross-metathesis reaction under mild, chemoselective and highly reproducible conditions. A systematic LC-MS/MS analysis of the reaction mixture allows locating unambigouslt the double bounds in fatty acid side chains. This method has en successfully applied at a nanomole scale to commerical standard mixtures as well as in lipid extracts from an in vitro model of corneal toxicity.<br><div><a></a><a></a><a></a><a></a></div><div><a></a><a></a><a></a> </div>

scholarly journals Exploratory Metabolomic Analysis Based on Reversed-Phase Liquid Chromatography–Mass Spectrometry to Study an In Vitro Model of Hypoxia-Induced Metabolic Alterations in HK-2 Cells

2021 ◽  
Vol 22 (14) ◽  
pp. 7399
Author(s):  
Samuel Bernardo-Bermejo ◽  
Elena Sánchez-López ◽  
Lei Tan ◽  
Selma Benito-Martínez ◽  
Zhengjin Jiang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
In Vitro Model ◽  
Reversed Phase ◽  
Liquid Chromatography Mass Spectrometry ◽  
Reversed Phase Liquid Chromatography ◽  
Metabolic Alterations ◽  
Phase Liquid ◽  
Vitro Model

Oxygen deficiency in cells, tissues, and organs can not only prevent the proper development of biological functions but it can also lead to several diseases and disorders. In this sense, the kidney deserves special attention since hypoxia can be considered an important factor in the pathophysiology of both acute kidney injury and chronic kidney disease. To provide better knowledge to unveil the molecular mechanisms involved, new studies are necessary. In this sense, this work aims to study, for the first time, an in vitro model of hypoxia-induced metabolic alterations in human proximal tubular HK-2 cells because renal proximal tubules are particularly susceptible to hypoxia. Different groups of cells, cultivated under control and hypoxia conditions at 0.5, 5, 24, and 48 h, were investigated using untargeted metabolomic approaches based on reversed-phase liquid chromatography–mass spectrometry. Both intracellular and extracellular fluids were studied to obtain a large metabolite coverage. On the other hand, multivariate and univariate analyses were carried out to find the differences among the cell groups and to select the most relevant variables. The molecular features identified as affected metabolites were mainly amino acids and Amadori compounds. Insights about their biological relevance are also provided.

An in vitro model for investigation of vitamin A effects on wound healing

2021 ◽  
pp. 1-6
Author(s):  
Hoda Keshmiri Neghab ◽  
Mohammad Hasan Soheilifar ◽  
Gholamreza Esmaeeli Djavid
Keyword(s):  
Wound Healing ◽  
Endothelial Cell ◽  
Vitamin A ◽  
In Vitro Model ◽  
Cellular Proliferation ◽  
Endothelial Cell Migration ◽  
Normal Fibroblast ◽  
Anti Inflammatory ◽  
Vitro Model

Abstract. Wound healing consists of a series of highly orderly overlapping processes characterized by hemostasis, inflammation, proliferation, and remodeling. Prolongation or interruption in each phase can lead to delayed wound healing or a non-healing chronic wound. Vitamin A is a crucial nutrient that is most beneficial for the health of the skin. The present study was undertaken to determine the effect of vitamin A on regeneration, angiogenesis, and inflammation characteristics in an in vitro model system during wound healing. For this purpose, mouse skin normal fibroblast (L929), human umbilical vein endothelial cell (HUVEC), and monocyte/macrophage-like cell line (RAW 264.7) were considered to evaluate proliferation, angiogenesis, and anti-inflammatory responses, respectively. Vitamin A (0.1–5 μM) increased cellular proliferation of L929 and HUVEC (p < 0.05). Similarly, it stimulated angiogenesis by promoting endothelial cell migration up to approximately 4 fold and interestingly tube formation up to 8.5 fold (p < 0.01). Furthermore, vitamin A treatment was shown to decrease the level of nitric oxide production in a dose-dependent effect (p < 0.05), exhibiting the anti-inflammatory property of vitamin A in accelerating wound healing. These results may reveal the therapeutic potential of vitamin A in diabetic wound healing by stimulating regeneration, angiogenesis, and anti-inflammation responses.

Präsidentenposter: Regulation der Expression der β-Defensine hBD-2 und hBD-3 in einem in vitro Model der Candida Ösophagitis: NF-κB, MAPK/AP-1 und EGFR

2009 ◽  
Vol 47 (09) ◽  
Author(s):  
N Steubesand ◽  
K Kiehne ◽  
R Pahl ◽  
G Brunke ◽  
UR Fölsch ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Vitro Model

A new in vitro model for pre-transplantation diagnosis of frozen/thawed human ovarian tissue

2011 ◽  
Vol 71 (05) ◽  
Author(s):  
M Salama ◽  
K Winkler ◽  
KF Murach ◽  
S Hofer ◽  
L Wildt ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Ovarian Tissue ◽  
Human Ovarian Tissue ◽  
Vitro Model
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