The Experts below are selected from a list of 1317 Experts worldwide ranked by ideXlab platform
Yuri L. Boteon - One of the best experts on this subject based on the ideXlab platform.
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manipulation of lipid metabolism during normothermic machine perfusion effect of Defatting therapies on donor liver functional recovery
Liver Transplantation, 2019Co-Authors: Yuri L. Boteon, Lorraine Wallace, Amanda P. C. S. Boteon, Joseph Attard, Gary M ReynoldsAbstract:BACKGROUND Strategies to increase steatotic donor livers' utilisation are required to tackle the mortality on the transplant waiting list. We aimed to test the efficacy of pharmacological enhancement of the lipid metabolism of human livers during ex-situ normothermic machine perfusion to promote Defatting and improve the functional recovery of the organs. METHODS Ten livers discarded because of steatosis were allocated to a Defatting group that had the perfusate supplemented with a combination of drugs to enhance lipid metabolism or a control group that received perfusion fluid with vehicle only. Steatosis was assessed using tissue homogenate and histological analysis. Markers for lipid oxidation and solubilisation, oxidative injury, inflammation and biliary function were evaluated by ELISA, immunohistochemistry and in-gel protein detection. RESULTS Treatment reduced tissue triglycerides by 38% and macrovesicular steatosis by 40% over 6 hours. This effect was driven by increased solubilisation of triglycerides (p=0.04); and mitochondrial oxidation as assessed by increased ketogenesis (p=0.008) and adenosine triphosphate synthesis (p=0.01) associated with raised levels of the enzymes ACOX-1, CPT1A and acetyl-CoA synthetase. Concomitantly, defatted livers exhibited enhanced metabolic functional parameters such as urea production (p=0.03), lower vascular resistance, lower release of alanine aminotransferase (p=0.049) and higher bile production (p=0.008) with a higher bile pH (p=0.03). The treatment downregulated the expression of markers for oxidative injury, activation of immune cells (CD-14; CD-11b) and reduced the release of inflammatory cytokines in the perfusate (TNF-α; IL-1β). CONCLUSION Pharmacological enhancement of intracellular lipid metabolism during normothermic machine perfusion decreased the lipid content of human livers within 6 hours. It also improved the intracellular metabolic support to the organs leading to successful functional recovery and decreased expression of markers of reperfusion injury. This article is protected by copyright. All rights reserved.
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Defatting of fat loaded primary human hepatocytes (PHH).
2018Co-Authors: Yuri L. Boteon, Lorraine Wallace, Amanda P. C. S. Boteon, Darius F. Mirza, Hynek Mergental, Ricky H. Bhogal, Simon AffordAbstract:Panel A: The positive are of oil red O of the Defatting treatment group was reduced by 28% in comparison with the vehicle control group over 24 hours and 54% over 48 hours. Panel B: Intracellular triglyceride levels of the Defatting treatment group were reduced by 32% within 24 hours of treatment and 35% within 48 hours, in comparison with the fatty vehicle control group. Panel C: Oil red O staining picture of PHH of the Defatting group at the end of the 48 hours of treatment. There is a predominance of small lipid droplets in the cytoplasm of the cells and the nucleus is in its usual position. Series 2: shows a series of oil red O staining pictures from PHH in culture at different time points of the experiments. Panel D shows lean cells in culture, after the incubation with fatty acids they become loaded with fat (panel E). Those fat loaded PHH were then incubated with only the vehicle of the drugs for 48 hours and the lipid content decreased over time (panel F) or had the Defatting treatment that showed the significant higher decrease in the area of oil red O (panel G). Data report the median of three separate experiments performed in quadruplicate and errors bars the interquartile range. Comparisons performed using two-tailed t-test. * = p
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Study design.
2018Co-Authors: Yuri L. Boteon, Lorraine Wallace, Amanda P. C. S. Boteon, Darius F. Mirza, Hynek Mergental, Ricky H. Bhogal, Simon AffordAbstract:Series 1: Isolated primary human hepatocytes (PHH) were left in standard media for 2 days and then received media supplemented with fatty acids. After 2 days of fat loading the fatty PHH were allocated to the Defatting treatment group where the media was supplemented with the Defatting cocktail of drugs, and the control groups, the standard control group and the vehicle control group that received vehicle only. Lean hepatocytes were kept in standard culture conditions throughout the experimental period. The experimentation period lasted for two days thereafter. Series 2: Human intra-hepatic endothelial cells (HIEC) and cholangiocytes were immuno-magnetically separated with Dynabeads conjugated with cell-specific monoclonal antibody. The cells were left in culture for 2 days in standard media to reach confluence and then were allocated to the intervention group that received the Defatting cocktail and the control groups, the standard control group and the vehicle control group that had the media supplemented with the vehicle only. The experimentation period lasts for two days thereafter.
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Assessment of the cytotoxicity of the Defatting cocktail to human cells of the liver via MTT assay.
2018Co-Authors: Yuri L. Boteon, Lorraine Wallace, Amanda P. C. S. Boteon, Darius F. Mirza, Hynek Mergental, Ricky H. Bhogal, Simon AffordAbstract:Panel A: the toxicity of the Defatting cocktail was tested in primary human hepatocytes and results showed a significant improvement of 11% in viability of the Defatting treatment group compared with the fatty vehicle control group. Panel B: Treatment of human intra-hepatic endothelial cells (HIEC) with the drugs had no effect on cell viability compared with the control groups. Panel C: treatment of cholangiocytes with the Defatting cocktail did not demonstrate any cytotoxic effect to the cells and indicated a slight improvement in viability compared to the control groups. Panels D and E: Phase contrast microscopy showing representative images of HIEC (Panel D) and cholangiocytes (Panel E) at different time points. No gross modifications in cell integrity were observed in either cell type which was consistent and supportive of the MTT data. Data report the median of three separate experiments performed in quadruplicate and errors bars the interquartile range. Comparisons performed using two-tailed t-test. * = p
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Release of total ketones in the supernatants.
2018Co-Authors: Yuri L. Boteon, Lorraine Wallace, Amanda P. C. S. Boteon, Darius F. Mirza, Hynek Mergental, Ricky H. Bhogal, Simon AffordAbstract:Fat loaded primary human hepatocytes that had the Defatting treatment showed an increase in cell culture supernatant levels of total ketone bodies of 1.22-fold over 24 hours and 1.40-fold over 48 hours. Data reports the median of three separate experiments performed in quadruplicate and errors bars the interquartile range. Comparisons performed using two-tailed t-test. * = p
Joseph Attard - One of the best experts on this subject based on the ideXlab platform.
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manipulation of lipid metabolism during normothermic machine perfusion effect of Defatting therapies on donor liver functional recovery
Liver Transplantation, 2019Co-Authors: Yuri L. Boteon, Lorraine Wallace, Amanda P. C. S. Boteon, Joseph Attard, Gary M ReynoldsAbstract:BACKGROUND Strategies to increase steatotic donor livers' utilisation are required to tackle the mortality on the transplant waiting list. We aimed to test the efficacy of pharmacological enhancement of the lipid metabolism of human livers during ex-situ normothermic machine perfusion to promote Defatting and improve the functional recovery of the organs. METHODS Ten livers discarded because of steatosis were allocated to a Defatting group that had the perfusate supplemented with a combination of drugs to enhance lipid metabolism or a control group that received perfusion fluid with vehicle only. Steatosis was assessed using tissue homogenate and histological analysis. Markers for lipid oxidation and solubilisation, oxidative injury, inflammation and biliary function were evaluated by ELISA, immunohistochemistry and in-gel protein detection. RESULTS Treatment reduced tissue triglycerides by 38% and macrovesicular steatosis by 40% over 6 hours. This effect was driven by increased solubilisation of triglycerides (p=0.04); and mitochondrial oxidation as assessed by increased ketogenesis (p=0.008) and adenosine triphosphate synthesis (p=0.01) associated with raised levels of the enzymes ACOX-1, CPT1A and acetyl-CoA synthetase. Concomitantly, defatted livers exhibited enhanced metabolic functional parameters such as urea production (p=0.03), lower vascular resistance, lower release of alanine aminotransferase (p=0.049) and higher bile production (p=0.008) with a higher bile pH (p=0.03). The treatment downregulated the expression of markers for oxidative injury, activation of immune cells (CD-14; CD-11b) and reduced the release of inflammatory cytokines in the perfusate (TNF-α; IL-1β). CONCLUSION Pharmacological enhancement of intracellular lipid metabolism during normothermic machine perfusion decreased the lipid content of human livers within 6 hours. It also improved the intracellular metabolic support to the organs leading to successful functional recovery and decreased expression of markers of reperfusion injury. This article is protected by copyright. All rights reserved.
Lorraine Wallace - One of the best experts on this subject based on the ideXlab platform.
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manipulation of lipid metabolism during normothermic machine perfusion effect of Defatting therapies on donor liver functional recovery
Liver Transplantation, 2019Co-Authors: Yuri L. Boteon, Lorraine Wallace, Amanda P. C. S. Boteon, Joseph Attard, Gary M ReynoldsAbstract:BACKGROUND Strategies to increase steatotic donor livers' utilisation are required to tackle the mortality on the transplant waiting list. We aimed to test the efficacy of pharmacological enhancement of the lipid metabolism of human livers during ex-situ normothermic machine perfusion to promote Defatting and improve the functional recovery of the organs. METHODS Ten livers discarded because of steatosis were allocated to a Defatting group that had the perfusate supplemented with a combination of drugs to enhance lipid metabolism or a control group that received perfusion fluid with vehicle only. Steatosis was assessed using tissue homogenate and histological analysis. Markers for lipid oxidation and solubilisation, oxidative injury, inflammation and biliary function were evaluated by ELISA, immunohistochemistry and in-gel protein detection. RESULTS Treatment reduced tissue triglycerides by 38% and macrovesicular steatosis by 40% over 6 hours. This effect was driven by increased solubilisation of triglycerides (p=0.04); and mitochondrial oxidation as assessed by increased ketogenesis (p=0.008) and adenosine triphosphate synthesis (p=0.01) associated with raised levels of the enzymes ACOX-1, CPT1A and acetyl-CoA synthetase. Concomitantly, defatted livers exhibited enhanced metabolic functional parameters such as urea production (p=0.03), lower vascular resistance, lower release of alanine aminotransferase (p=0.049) and higher bile production (p=0.008) with a higher bile pH (p=0.03). The treatment downregulated the expression of markers for oxidative injury, activation of immune cells (CD-14; CD-11b) and reduced the release of inflammatory cytokines in the perfusate (TNF-α; IL-1β). CONCLUSION Pharmacological enhancement of intracellular lipid metabolism during normothermic machine perfusion decreased the lipid content of human livers within 6 hours. It also improved the intracellular metabolic support to the organs leading to successful functional recovery and decreased expression of markers of reperfusion injury. This article is protected by copyright. All rights reserved.
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Defatting of fat loaded primary human hepatocytes (PHH).
2018Co-Authors: Yuri L. Boteon, Lorraine Wallace, Amanda P. C. S. Boteon, Darius F. Mirza, Hynek Mergental, Ricky H. Bhogal, Simon AffordAbstract:Panel A: The positive are of oil red O of the Defatting treatment group was reduced by 28% in comparison with the vehicle control group over 24 hours and 54% over 48 hours. Panel B: Intracellular triglyceride levels of the Defatting treatment group were reduced by 32% within 24 hours of treatment and 35% within 48 hours, in comparison with the fatty vehicle control group. Panel C: Oil red O staining picture of PHH of the Defatting group at the end of the 48 hours of treatment. There is a predominance of small lipid droplets in the cytoplasm of the cells and the nucleus is in its usual position. Series 2: shows a series of oil red O staining pictures from PHH in culture at different time points of the experiments. Panel D shows lean cells in culture, after the incubation with fatty acids they become loaded with fat (panel E). Those fat loaded PHH were then incubated with only the vehicle of the drugs for 48 hours and the lipid content decreased over time (panel F) or had the Defatting treatment that showed the significant higher decrease in the area of oil red O (panel G). Data report the median of three separate experiments performed in quadruplicate and errors bars the interquartile range. Comparisons performed using two-tailed t-test. * = p
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Study design.
2018Co-Authors: Yuri L. Boteon, Lorraine Wallace, Amanda P. C. S. Boteon, Darius F. Mirza, Hynek Mergental, Ricky H. Bhogal, Simon AffordAbstract:Series 1: Isolated primary human hepatocytes (PHH) were left in standard media for 2 days and then received media supplemented with fatty acids. After 2 days of fat loading the fatty PHH were allocated to the Defatting treatment group where the media was supplemented with the Defatting cocktail of drugs, and the control groups, the standard control group and the vehicle control group that received vehicle only. Lean hepatocytes were kept in standard culture conditions throughout the experimental period. The experimentation period lasted for two days thereafter. Series 2: Human intra-hepatic endothelial cells (HIEC) and cholangiocytes were immuno-magnetically separated with Dynabeads conjugated with cell-specific monoclonal antibody. The cells were left in culture for 2 days in standard media to reach confluence and then were allocated to the intervention group that received the Defatting cocktail and the control groups, the standard control group and the vehicle control group that had the media supplemented with the vehicle only. The experimentation period lasts for two days thereafter.
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Assessment of the cytotoxicity of the Defatting cocktail to human cells of the liver via MTT assay.
2018Co-Authors: Yuri L. Boteon, Lorraine Wallace, Amanda P. C. S. Boteon, Darius F. Mirza, Hynek Mergental, Ricky H. Bhogal, Simon AffordAbstract:Panel A: the toxicity of the Defatting cocktail was tested in primary human hepatocytes and results showed a significant improvement of 11% in viability of the Defatting treatment group compared with the fatty vehicle control group. Panel B: Treatment of human intra-hepatic endothelial cells (HIEC) with the drugs had no effect on cell viability compared with the control groups. Panel C: treatment of cholangiocytes with the Defatting cocktail did not demonstrate any cytotoxic effect to the cells and indicated a slight improvement in viability compared to the control groups. Panels D and E: Phase contrast microscopy showing representative images of HIEC (Panel D) and cholangiocytes (Panel E) at different time points. No gross modifications in cell integrity were observed in either cell type which was consistent and supportive of the MTT data. Data report the median of three separate experiments performed in quadruplicate and errors bars the interquartile range. Comparisons performed using two-tailed t-test. * = p
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Release of total ketones in the supernatants.
2018Co-Authors: Yuri L. Boteon, Lorraine Wallace, Amanda P. C. S. Boteon, Darius F. Mirza, Hynek Mergental, Ricky H. Bhogal, Simon AffordAbstract:Fat loaded primary human hepatocytes that had the Defatting treatment showed an increase in cell culture supernatant levels of total ketone bodies of 1.22-fold over 24 hours and 1.40-fold over 48 hours. Data reports the median of three separate experiments performed in quadruplicate and errors bars the interquartile range. Comparisons performed using two-tailed t-test. * = p
Gary M Reynolds - One of the best experts on this subject based on the ideXlab platform.
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manipulation of lipid metabolism during normothermic machine perfusion effect of Defatting therapies on donor liver functional recovery
Liver Transplantation, 2019Co-Authors: Yuri L. Boteon, Lorraine Wallace, Amanda P. C. S. Boteon, Joseph Attard, Gary M ReynoldsAbstract:BACKGROUND Strategies to increase steatotic donor livers' utilisation are required to tackle the mortality on the transplant waiting list. We aimed to test the efficacy of pharmacological enhancement of the lipid metabolism of human livers during ex-situ normothermic machine perfusion to promote Defatting and improve the functional recovery of the organs. METHODS Ten livers discarded because of steatosis were allocated to a Defatting group that had the perfusate supplemented with a combination of drugs to enhance lipid metabolism or a control group that received perfusion fluid with vehicle only. Steatosis was assessed using tissue homogenate and histological analysis. Markers for lipid oxidation and solubilisation, oxidative injury, inflammation and biliary function were evaluated by ELISA, immunohistochemistry and in-gel protein detection. RESULTS Treatment reduced tissue triglycerides by 38% and macrovesicular steatosis by 40% over 6 hours. This effect was driven by increased solubilisation of triglycerides (p=0.04); and mitochondrial oxidation as assessed by increased ketogenesis (p=0.008) and adenosine triphosphate synthesis (p=0.01) associated with raised levels of the enzymes ACOX-1, CPT1A and acetyl-CoA synthetase. Concomitantly, defatted livers exhibited enhanced metabolic functional parameters such as urea production (p=0.03), lower vascular resistance, lower release of alanine aminotransferase (p=0.049) and higher bile production (p=0.008) with a higher bile pH (p=0.03). The treatment downregulated the expression of markers for oxidative injury, activation of immune cells (CD-14; CD-11b) and reduced the release of inflammatory cytokines in the perfusate (TNF-α; IL-1β). CONCLUSION Pharmacological enhancement of intracellular lipid metabolism during normothermic machine perfusion decreased the lipid content of human livers within 6 hours. It also improved the intracellular metabolic support to the organs leading to successful functional recovery and decreased expression of markers of reperfusion injury. This article is protected by copyright. All rights reserved.
Heidi Yeh - One of the best experts on this subject based on the ideXlab platform.
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improvement of steatotic rat liver function with a Defatting cocktail during ex situ normothermic machine perfusion is not directly related to liver fat content
PLOS ONE, 2020Co-Authors: Siavash Raigani, Martin L. Yarmush, Cailah Carroll, Stephanie Griffith, Casie A Pendexter, Ivy A Rosales, Hany Deirawan, Rafic Beydoun, Korkut Uygun, Heidi YehAbstract:There is a significant organ shortage in the field of liver transplantation, partly due to a high discard rate of steatotic livers from donors. These organs are known to function poorly if transplanted but make up a significant portion of the available pool of donated livers. This study demonstrates the ability to improve the function of steatotic rat livers using a combination of ex situ machine perfusion and a "Defatting" drug cocktail. After 6 hours of perfusion, defatted livers demonstrated lower perfusate lactate levels and improved bile quality as demonstrated by higher bile bicarbonate and lower bile lactate. Furthermore, Defatting was associated with decreased gene expression of pro-inflammatory cytokines and increased expression of enzymes involved in mitochondrial fatty acid oxidation. Rehabilitation of marginal or discarded steatotic livers using machine perfusion and tailored drug therapy can significantly increase the supply of donor livers for transplantation.
