scholarly journals Oleate prevents palmitate-induced mitochondrial dysfunction in chondrocytes

2020 ◽  
Author(s):  
Maria Vaquez-Mosquera ◽  
Mercedes Fernandez-Moreno ◽  
Estefania Cortes-Pereira ◽  
Sara Relaño ◽  
Andrea Dalmao-Fernandez ◽  
...  
Keyword(s):  
Mitochondrial Dysfunction ◽  
Lipid Droplets ◽  
Coupling Efficiency ◽  
Human Chondrocytes ◽  
Metabolic Phenotype ◽  
Systemic Factors ◽  
Atp Bioluminescence ◽  
Bioluminescence Assay ◽  
Atp Bioluminescence Assay

Abstract BACKGROUND : The clear association between obesity and osteoarthritis (OA) in joints not subjected to mechanical overload, together with the relationship between OA and metabolic syndrome (MS), suggests that there are systemic factors related to metabolic disorders that are involved in the metabolic phenotype of OA. The aim of this work is to study the effects of palmitate (PA) and oleate (OL), as the most abundant fatty acids (FA) present in the diet and serum, on cellular metabolism in an " in vitro " model of human chondrocytes. METHODS :.The Seahorse XF96 Analyzer was used tomeasure the mitochondrial, glycolytic function and the contribution of the mitochondrial oxidative phosphorilation system (OXPHOS) and glycolysis to the production of ATP, in the T/C-28a2 chondrocyte treated with to PA, OL and palmitate/oleate (PA/OL) ratio 1:2. Subsequently ATP bioluminescence assay kit was used for ATP quantification. To detect the presence of lipid droplets, two types of stains were performed and the amount of Triglycerides was quantified spectrophotometrically. RESULTS : PA, but not OL, produces mitochondrial dysfunction observed with a lower rate of OCR intended for the synthesis of ATP, coupling efficiency, maximal respiration and spare respiratory capacity. Glycolytic function showed lower rates for both glycolytic capacity and glycolytic reserve when cells were incubated withFA in relation to basal condition (BC). The production rate of ATP from OXPHOS showed lower values in chondrocytes incubated with any of the FAs. The evaluation of possible formation of Lipid droplets (LD) showed a significant increase of these structures in FA conditions, being significantly higher when the cells were incubated with OL. CONCLUSIONS : PA and OL show antagonistic effects in human chondrocytes; while increased levels of PA induce mitochondrial dysfunction and hinder the response of chondrocytes through the glycolytic pathway, OL shows a cytoprotective effect through which promotes the formation of triglycerides-rich LD, as well as the incorporation of PA.

scholarly journals Oleate Prevents Palmitate-Induced Mitochondrial Dysfunction in Chondrocytes

2021 ◽  
Vol 12 ◽  
Author(s):  
Maria Eugenia Vázquez-Mosquera ◽  
Mercedes Fernández-Moreno ◽  
Estefanía Cortés-Pereira ◽  
Sara Relaño ◽  
Andrea Dalmao-Fernández ◽  
...  
Keyword(s):  
Mitochondrial Dysfunction ◽  
Lipid Droplets ◽  
Coupling Efficiency ◽  
Metabolic Phenotype ◽  
Atp Production ◽  
Close Relationship ◽  
Systemic Factors ◽  
Relationship Of ◽  
The Relationship

The association between obesity and osteoarthritis (OA) in joints not subjected to mechanical overload, together with the relationship between OA and metabolic syndrome, suggests that there are systemic factors related to metabolic disorders that are involved in the metabolic phenotype of OA. The aim of this work is study the effects of palmitate and oleate on cellular metabolism in an “in vitro” model of human chondrocytes. The TC28a2 chondrocyte cell line was used to analyze the effect of palmitate and oleate on mitochondrial and glycolytic function, Adenosine triphosphate (ATP) production and lipid droplets accumulation. Palmitate, but not oleate, produces mitochondrial dysfunction observed with a lower coupling efficiency, maximal respiration and spare respiratory capacity. Glycolytic function showed lower rates both glycolytic capacity and glycolytic reserve when cells were incubated with fatty acids (FAs). The production rate of total and mitochondrial ATP showed lower values in chondrocytes incubated with palmitic acid (PA). The formation of lipid droplets increased in FA conditions, being significantly higher when the cells were incubated with oleic acid (OL). These results may help explain, at least in part, the close relationship of metabolic pathologies with OA, as well as help to elucidate some of the factors that can define a metabolic phenotype in OA.

scholarly journals Rapid Antibiotic Combination Testing for Carbapenem-Resistant Gram-Negative Bacteria within Six Hours Using ATP Bioluminescence

2018 ◽  
Vol 62 (9) ◽  
Author(s):  
Yiying Cai ◽  
Celene L. Seah ◽  
Hui Leck ◽  
Tze-Peng Lim ◽  
Jocelyn Q. Teo ◽  
...  
Keyword(s):  
Predictive Accuracy ◽  
Roc Curves ◽  
Gram Negative Bacteria ◽  
Content Type ◽  
Carbapenem Resistant ◽  
Atp Bioluminescence ◽  
Bioluminescence Assay ◽  
Atp Bioluminescence Assay ◽  
Selection Of

ABSTRACTTo guide the timely selection of antibiotic combinations against carbapenem-resistant Gram-negative bacteria (CR-GNB), anin vitrotest with a short turnaround time is essential. We developed anin vitroATP bioluminescence assay to determine effective antibiotic combinations against CR-GNB within 6 h. We tested 42 clinical CR-GNB strains (14Acinetobacter baumannii, 14Pseudomonas aeruginosa, and 14Klebsiella pneumoniaestrains) against 74 single antibiotics and two-antibiotic combinations. Bacteria (approximately 5 log10CFU/ml) were incubated with an antibiotic(s) at 35°C; ATP bioluminescence was measured at 6 h and 24 h; and the measurements were compared to viable counts at 24 h. Receiver operating characteristic (ROC) curves were used to determine the optimal luminescence thresholds (TRLU) for distinguishing between inhibitory and noninhibitory combinations. The areas under the 6-h and 24-h ROC curves were compared using the DeLong method. Prospective validation of the established thresholds was conducted using 18 additional CR-GNB. The predictive accuracy ofTRLUfor the 6-h ATP bioluminescence assay was 77.5% when all species were analyzed collectively. Predictive accuracies ranged from 73.7% to 82.7% when each species was analyzed individually. Upon comparison of the areas under the 6-h and 24-h ROC curves, the 6-h assay performed significantly better than the 24-h assay (P< 0.01). Predictive accuracy remained high upon prospective validation of the 6-h ATP assay (predictive accuracy, 79.8%; 95% confidence interval [CI], 77.6 to 81.9%), confirming the external validity of the assay. Our findings indicate that our 6-h ATP bioluminescence assay can provide guidance for prospective selection of antibiotic combinations against CR-GNB in a timely manner and may be useful in the management of CR-GNB infections.

Use of a Rapid Microbial ATP Bioluminescence Assay to Detect Contamination on Beef and Pork Carcasses†

1995 ◽  
Vol 58 (7) ◽  
pp. 770-775 ◽  
Author(s):  
GREGORY R. SIRAGUSA ◽  
CATHERINE N. CUTTER ◽  
WARREN J. DORSA ◽  
MOHAMMAD KOOHMARAIE
Keyword(s):  
Plate Count ◽  
Assay Sensitivity ◽  
Standard Plate Count ◽  
Critical Control Points ◽  
Processing Plants ◽  
Atp Bioluminescence ◽  
Bioluminescence Assay ◽  
Commercial Processing ◽  
Atp Bioluminescence Assay

A new microbial ATP bioluminescence assay was shown to be an accurate and rapid method to determine the levels of generic bacterial contamination on beef (n = 400 and pork (n = 320) carcasses sampled in commercial processing plants. Based on in vitro fecal dilution studies, the rapid microbial ATP (R-mATP) assay is as accurate as the standard plate count method for estimating bacteria in bovine or porcine fecal samples. The correlations (r) between the R-mATP assay and the standard aerobic plate count for beef and pork carcasses sampled in commercial processing were 0.91 and 0.93, respectively. A segmented-model statistical approach to determine the lower limits of assay sensitivity was developed. By using this model to analyze the in-plant data, the R-mATP test responded in a linear fashion to levels of microbial contamination of &gt; log10 2.0 aerobic CFU/cm2 on beef carcasses and of &gt; log10 3.2 aerobic CFU/cm2 for pork carcasses. The R-mATP assay requires approximately 5 min to complete, including sampling. Given the rapidity and accuracy of the assay, processors interested in monitoring critical control points in the slaughter process could potentially use the R-mATP assay to monitor microbiological prevention and intervention procedures for minimizing carcass contamination.

scholarly journals ATP-bioluminescence assay as an alternative for hygiene-monitoring procedures of stainless steel milk contact surfaces

2006 ◽  
Vol 37 (3) ◽  
pp. 345-349 ◽  
Author(s):  
Patrícia Dolabela Costa ◽  
Nélio José Andrade ◽  
Sebastião César Cardoso Brandão ◽  
Frederico José Vieira Passos ◽  
Nilda de Fátima Ferreira Soares
Keyword(s):  
Stainless Steel ◽  
Atp Bioluminescence ◽  
Bioluminescence Assay ◽  
Contact Surfaces ◽  
Atp Bioluminescence Assay

Detection of fungi and control of disinfection by ATP-bioluminescence assay

2003 ◽  
Vol 28 (1) ◽  
pp. 16-22 ◽  
Author(s):  
Malalanirina Rakotonirainy ◽  
Jozef Hanus ◽  
Sylvette Bonassies-Termes ◽  
Cécile Heraud ◽  
Bertrand Lavédrine
Keyword(s):  
Atp Bioluminescence ◽  
Bioluminescence Assay ◽  
Atp Bioluminescence Assay ◽  
And Control
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