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https://hdl.handle.net/10171/7301
2024-03-28T23:49:46ZMonitoring Caenorhabditis elegans molting in a conventional luminometer
https://hdl.handle.net/10171/69226
Title: Monitoring Caenorhabditis elegans molting in a conventional luminometer
Abstract: Molting is an essential developmental process in Caenorhabditis elegans. However, the study of
molting in the worm has been limited by the lack of automated techniques that allow monitoring
the process in a simple way. In 2015, Olmedo et al. published an automated method to monitor
the timing of each larval stage and molt in C. elegans using bioluminescence. This new method
has greatly contributed to the study of molting in this organism but requires the use of a highsensitivity luminometer, which many laboratories do not have. We have adapted the method to a
conventional luminometer, so that it can be used by most laboratories that work with C. elegans
and do not have high-sensitivity equipment.
• A customization of a method to study molting in C. elegans using a conventional luminometer
instead of a high-sensitivity one.
• This adaptation allows most laboratories to use their routine luminometers to study molting
in C. elegans.
• Although the use of a high-sensitivity luminometer, as proposed by Olmedo et al., remains
the gold standard for studying molting, this adaptation is suitable for studying significant
differences in molting and the duration of larval stages between different strains of C. elegans.2023-01-01T00:00:00ZAntifibrogenic and apoptotic effects of Ocoxin in cultured rat hepatic stellate cells
https://hdl.handle.net/10171/69211
Title: Antifibrogenic and apoptotic effects of Ocoxin in cultured rat hepatic stellate cells
Abstract: Ocoxin is a nutritional supplement that has been shown to exert antioxidant and immunomodulatory responses in patients
with chronic hepatitis C. The present work aimed to determine the efects of Ocoxin on activated hepatic stellate cells (HSC),
the cell type mainly responsible for collagen deposition in the fbrotic liver. Ocoxin was found to reduce the survival of a
cell line of immortalized non-tumoral rat HSC in a dose–response fashion and to diminish collagen type I levels. This latter
efect was observed even at doses not afecting cell survival, pointing to an antifbrogenic action for the supplement. The
decrease in viability exerted by Ocoxin on HSC correlated with an increase in histone-associated fragments in the cytoplasm
and with increased activity of caspase-3, indicating the induction of apoptosis. To determine the molecular mechanisms
mediating Ocoxin-induced apoptosis, the activation of members of the MAPK family was analyzed. Incubation of HSC
with Ocoxin caused a transient and dramatic enhancement on ERK, JNK, and p38 MAPK phosphorylation levels. Using
specifc inhibitors for these enzymes, p38 MAPK was identifed as a key mediator of the apoptotic efect of Ocoxin on HSC.2022-01-01T00:00:00ZEndothelial NOX5 overexpression induces changes in the cardiac gene profile: potential impact in myocardial infarction?
https://hdl.handle.net/10171/69202
Title: Endothelial NOX5 overexpression induces changes in the cardiac gene profile: potential impact in myocardial infarction?
Abstract: Cardiovascular diseases and the ischemic heart disease specifically constitute the main cause of death worldwide. The ischemic heart disease may lead to myocardial infarction, which in turn triggers numerous mechanisms and pathways involved in cardiac repair and remodeling. Our goal in the present study was to characterize the effect of the NADPH oxidase 5 (NOX5) endothelial expression in healthy and infarcted knock-in mice on diverse signaling pathways. The mechanisms studied in the heart of mice were the redox pathway, metalloproteinases and collagen pathway, signaling factors such as NFκB, AKT or Bcl-2, and adhesion molecules among others. Recent studies support that NOX5 expression in animal models can modify the environment and predisposes organ response to harmful stimuli prior to pathological processes. We found many alterations in the mRNA expression of components involved in cardiac fibrosis as collagen type I or TGF-β and in key players of cardiac apoptosis such as AKT, Bcl-2, or p53. In the heart of NOX5-expressing mice after chronic myocardial infarction, gene alterations were predominant in the redox pathway (NOX2, NOX4, p22phox, or SOD1), but we also found alterations in VCAM-1 and β-MHC expression. Our results suggest that NOX5 endothelial expression in mice preconditions the heart, and we propose that NOX5 has a cardioprotective role. The correlation studies performed between echocardiographic parameters and cardiac mRNA expression supported NOX5 protective action.2023-01-01T00:00:00ZMatrix metalloproteinase-10 deficiency delays atherosclerosis progression and plaque calcification
https://hdl.handle.net/10171/68977
Title: Matrix metalloproteinase-10 deficiency delays atherosclerosis progression and plaque calcification
Abstract: Background and aims: Matrix metalloproteinases (MMPs) have been implicated in atherosclerosis and vascular calcification. Among them, we reported that MMP10 is present in human atheroma, associated with atherosclerosis. However, it remains unclear whether MMP10 is involved in atherogenesis and vascular calcification.
Methods: MMP10 was measured in serum from patients with subclinical atherosclerosis and analyzed in carotid endarterectomies by immunostaining. ApoE-deficient mice (Apoe-/-) were crossed to MMP10-deficient (Mmp10-/-) mice and followed up to 20 months. Plaque area and composition were assessed by histology and immunohistochemistry. Inflammatory markers were measured in atherosclerotic plaques by RT-qPCR, and leukocyte subpopulations were analyzed by flow cytometry. In vitro calcification assays were performed in aortic vascular smooth muscle cells (VSMC).
Results: MMP10 serum levels were associated with coronary calcification in subjects with subclinical atherosclerosis. Immunostaining revealed MMP10 expression in human atheromas, spatially associated with calcification areas, and complicated plaques released higher amounts of MMP10 than non-diseased segments. Interestingly, vascular MMP10 expression was confined to the atherosclerotic lesion in Apoe-/- mice, and Apoe-/-Mmp10-/- showed a substantial reduction in atherosclerotic lesion size, macrophage content and plaque calcification. Reduced local and systemic inflammatory markers could be demonstrated in Apoe-/-Mmp10-/- by gene expression and flow cytometry analysis. Calcium phosphate deposition and vascular calcification markers were downregulated in VSMC from Apoe-/-Mmp10-/- mice.
Conclusions: Delayed plaque progression and altered cellular composition in the absence of MMP10 suggests that MMP10 plays a role in atherosclerosis, favoring inflammation, development and complication of the plaque.2018-01-01T00:00:00Z