scholarly journals The Effect of Rivaroxaban on CYP4F2 and Transcription Factors’ Activity in HUVECs

2021 ◽  
Vol 11 (22) ◽  
pp. 10851
Author(s):  
Ieva Ciapiene ◽  
Vacis Tatarunas ◽  
Agne Giedraitiene ◽  
Vaidotas Zvikas ◽  
Valdas Jakstas ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Transcription Factors ◽  
Protein Concentration ◽  
Culture Medium ◽  
Hepatic Metabolism ◽  
Esi Ms ◽  
Protein Levels ◽  
M 10 ◽  
The Impact

Interindividual variabilities between patients taking the anticoagulant rivaroxaban are a result of hepatic metabolism by CYP 450 enzymes. The objective of this study was to evaluate the impact of rivaroxaban on CYP4F2 and transcription factors’ activity in HUVECs. Rivaroxaban and its metabolites were detected by UPLC-ESI-MS and UPLC-QTOF-MS. CYP4F2, HNF4α, PXR and CAR expressions were determined in HUVECs by qPCR; CYP4F2 protein concentration was determined by ELISA. Rivaroxaban metabolites (M-1, M-2, M-5, M-8, M-10, M-11 and M-18) were detected in endothelial cells’ culture medium. Increasing concentrations of rivaroxaban determined lower 13-docosenamide concentrations. Rivaroxaban and dexamethasone reduced the expression of CYP4F2 when hsa-miR-24-3p—both CYP4F2 expression and CYP4F2 protein levels in HUVECs. The expression of the transcription factors HNF4α, PXR and CAR was not detected in HUVECs.

scholarly journals Mitochondria maintain maturation and secretion of lipoprotein lipase in the endoplasmic reticulum

2006 ◽  
Vol 396 (1) ◽  
pp. 173-182 ◽  
Author(s):  
Karin Osibow ◽  
Sasa Frank ◽  
Roland Malli ◽  
Rudolf Zechner ◽  
Wolfgang F. Graier
Keyword(s):  
Endothelial Cells ◽  
Endoplasmic Reticulum ◽  
Protein Folding ◽  
Lipoprotein Lipase ◽  
Chinese Hamster ◽  
Protein Levels ◽  
Dependent Activity ◽  
Dependent Protein ◽  
The Impact ◽  
Reversible Reduction

Considering the physiological Ca2+ dynamics within the ER (endoplasmic reticulum), it remains unclear how efficient protein folding is maintained in living cells. Thus, utilizing the strictly folding-dependent activity and secretion of LPL (lipoprotein lipase), we evaluated the impact of ER Ca2+ content and mitochondrial contribution to Ca2+-dependent protein folding. Exhaustive ER Ca2+ depletion by inhibition of sarcoplasmic/endoplasmic reticulum Ca2+-ATPases caused strong, but reversible, reduction of cell-associated and released activity of constitutive and adenovirus-encoded human LPL in CHO-K1 (Chinese-hamster ovary K1) and endothelial cells respectively, which was not due to decline of mRNA or intracellular protein levels. In contrast, stimulation with the IP3 (inositol 1,4,5-trisphosphate)-generating agonist histamine only moderately and transiently affected LPL maturation in endothelial cells that paralleled a basically preserved ER Ca2+ content. However, in the absence of extracellular Ca2+ or upon prevention of transmitochondrial Ca2+ flux, LPL maturation discontinued upon histamine stimulation. Collectively, these data indicate that Ca2+-dependent protein folding in the ER is predominantly controlled by intraluminal Ca2+ and is largely maintained during physiological cell stimulation owing to efficient ER Ca2+ refilling. Since Ca2+ entry and mitochondrial Ca2+ homoeostasis are crucial for continuous Ca2+-dependent protein maturation in the ER, their pathological alterations may result in dysfunctional protein folding.

scholarly journals A method for differentiating human induced pluripotent stem cells toward functional cardiomyocytes in 96-well microplates

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Novin Balafkan ◽  
Sepideh Mostafavi ◽  
Manja Schubert ◽  
Richard Siller ◽  
Kristina Xiao Liang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Small Molecules ◽  
Pluripotent Stem Cells ◽  
Culture Medium ◽  
Electrophysiological Properties ◽  
Human Cardiomyocytes ◽  
Protein Levels ◽  
Incremental Improvement ◽  
Induced Pluripotent ◽  
The Impact

Abstract The capacity of pluripotent stem cells both for self-renewal and to differentiate into any cell type have made them a powerful tool for studying human disease. Protocols for efficient differentiation towards cardiomyocytes using defined, serum-free culture medium combined with small molecules have been developed, but thus far, limited to larger formats. We adapted protocols for differentiating human pluripotent stem cells to functional human cardiomyocytes in a 96-well microplate format. The resulting cardiomyocytes expressed cardiac specific markers at the transcriptional and protein levels and had the electrophysiological properties that confirmed the presence of functional cardiomyocytes. We suggest that this protocol provides an incremental improvement and one that reduces the impact of heterogeneity by increasing inter-experimental replicates. We believe that this technique will improve the applicability of these cells for use in developmental biology and mechanistic studies of disease.

scholarly journals Cell Type Dependent Suppression of Inflammatory Mediators by Myocardin Related Transcription Factors

2021 ◽  
Vol 12 ◽  
Author(s):  
Li Liu ◽  
Elisabeth Bankell ◽  
Catarina Rippe ◽  
Björn Morén ◽  
Karin G. Stenkula ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Transcription Factors ◽  
Vascular Inflammation ◽  
Arterial Disease ◽  
Suppressive Effect ◽  
Cell Type ◽  
Human Coronary Artery ◽  
Anti Inflammatory ◽  
The Impact

Myocardin related transcription factors (MRTFs: MYOCD/myocardin, MRTF-A, and MRTF-B) play a key role in smooth muscle cell differentiation by activating contractile genes. In atherosclerosis, MRTF levels change, and most notable is a fall of MYOCD. Previous work described anti-inflammatory properties of MRTF-A and MYOCD, occurring through RelA binding, suggesting that MYOCD reduction could contribute to vascular inflammation. Recent studies have muddled this picture showing that MRTFs may show both anti- and pro-inflammatory properties, but the basis of these discrepancies remain unclear. Moreover, the impact of MRTFs on inflammatory signaling pathways in tissues relevant to human arterial disease is uncertain. The current work aimed to address these issues. RNA-sequencing after forced expression of myocardin in human coronary artery smooth muscle cells (hCASMCs) showed reduction of pro-inflammatory transcripts, including CCL2, CXCL8, IL6, and IL1B. Side-by-side comparison of MYOCD, MRTF-A, and MRTF-B in hCASMCs, showed that the anti-inflammatory impact was shared among MRTFs. Correlation analyses using human arterial transcriptomic datasets revealed negative correlations between MYOCD, MRTFA, and SRF, on the one hand, and the inflammatory transcripts, on the other. A pro-inflammatory drive from lipopolysaccharide, did not change the size of the suppressive effect of MRTF-A in hCASMCs on either mRNA or protein levels. To examine cell type-dependence, we compared the anti-inflammatory impact in hCASMCs, with that in human bladder SMCs, in endothelial cells, and in monocytes (THP-1 cells). Surprisingly, little anti-inflammatory activity was seen in endothelial cells and monocytes, and in bladder SMCs, MRTF-A was pro-inflammatory. CXCL8, IL6, and IL1B were increased by the MRTF-SRF inhibitor CCG-1423 and by MRTF-A silencing in hCASMCs, but depolymerization of actin, known to inhibit MRTF activity, had no stimulatory effect, an exception being IL1B. Co-immunoprecipitation supported binding of MRTF-A to RelA, supporting sequestration of this important pro-inflammatory mediator as a mechanism. Dexamethasone treatment and silencing of RelA (by 76 ± 1%) however only eliminated a fraction of the MRTF-A effect (≈25%), suggesting mechanisms beyond RelA binding. Indeed, SRF silencing suggested that MRTF-A suppression of IL1B and CXCL8 depends on SRF. This work thus supports an anti-inflammatory impact of MRTF-SRF signaling in hCASMCs and in intact human arteries, but not in several other cell types.

Angiogenesis Is Suppressed by Junctional Adhesion Molecule A Through Transcription Factors HIF-1a- and Id1-Dependent Modulation of VEGF/VEGFR2 Expression

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 618-618
Author(s):  
Meghna Ulhas Naik ◽  
Vesselina Cooke ◽  
Sharmila Chatterjee ◽  
Ulhas P Naik
Keyword(s):  
Endothelial Cells ◽  
Transcription Factors ◽  
Growth Factor ◽  
Tumor Growth ◽  
Vascular Permeability ◽  
Adhesion Molecule ◽  
Vascular Endothelial ◽  
Protein Levels ◽  
Murine Melanoma ◽  
Signaling Axis

Abstract Abstract 618 The process of angiogenesis is associated with a number of human pathologies. Vascular endothelial growth factor (VEGF) is a potent angiogenic growth factor. VEGF expression is upregulated during ischemia and tumor growth. VEGF is also expressed in healthy adult vascular tissue. However, VEGF-dependent angiogenesis is normally suppressed in healthy adults. The mechanism of this suppression is not known. Here we show that junctional adhesion molecule A (JAM-A), a tight junction protein, is an endogenous suppressor of VEGF-induced angiogenesis. Using gene-targeted Jam-A null mice, we found that murine melanoma tumor growth and associated angiogenesis were significantly augmented (P<0.001) in Jam-A null mice compared to wild-type mice. Additionally, Jam-A null mice showed significantly enhanced (P<0.004) vascular permeability as assessed by a Miles assay using Evans blue dye. Furthermore, VEGF-, but not FGF-2-dependent angiogenesis is significantly augmented (P<0.001) in the absence of Jam-A as assessed by Matrigel plug and aortic ring sprouting, two well-accepted in vivo and ex vivo angiogenic assays, respectively. Vascular endothelial cells isolated from Jam-A null mouse aorta showed significantly enhanced (P<0.05) cell migration and tube-like structure formation (P<0.05) in response to VEGF. Additionally, we found the plasma levels of VEGF in Jam-A null mice is significantly (P<0.000001) and age-dependently increased compared to WT mice. Furthermore, both mRNA and protein levels of VEGF and its receptor VEGFR2 were significantly increased (P<0.001) in Jam-A null endothelial cells, suggesting that the VEGF/VEGFR2 signaling axis is enhanced in the absence of Jam-A. To further confirm this finding, we injected anti-VEGFR2 (DC101) into the Jam-A null mice inoculated with murine melanoma (B16F0) cells. Inhibition of VEGF/VEGFR-2 signaling by DC101 significantly reduced (P<0.00003) tumor growth and associated angiogenesis in Jam-A null mice. Additionally, vascular permeability observed in Jam-A null mice was completely abrogated upon DC101 treatment. When tested if the expression of soluble Flt (sFlt), which is known to trap VEGF, is downregulated in the absence of Jam-A, we found no significant difference in sFlt mRNA expression in Jam-A null endothelial cells compared to WT. In order to determine the mechanism of this upregulation of the VEGF signaling axis, we tested the expression of hypoxia inducible factor 1a (HIF1-a) and inhibitor of DNA binding 1 (Id1), two transcription factors known to upregulate VEGF and VEGFR2 gene expression. Interestingly, mRNA and protein levels of both HIF1-a and Id1 were significantly augmented (P<0.02) in endothelial cells lacking Jam-A. Consistent with this finding, the overexpression of JAM-A in HUVECs attenuated the levels of Id1. These results suggest that JAM-A suppresses VEGF/VEGFR2 expression in endothelial cells by attenuating HIF1-a and Id1 expression, thus suppressing adult angiogenesis. During pathological conditions such as ischemia and tumor growth, it is possible that JAM-A levels are downregulated, thus supporting pathological angiogenesis. Disclosures: No relevant conflicts of interest to declare.

PTEN expression in endothelial cells is down-regulated by uPAR to promote angiogenesis

2015 ◽  
Vol 114 (08) ◽  
pp. 379-389 ◽  
Author(s):  
Matthias Unseld ◽  
Anastasia Chilla ◽  
Clemens Pausz ◽  
Rula Mawas ◽  
Johannes Breuss ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
In Vitro Assays ◽  
Soluble Form ◽  
Dependent Manner ◽  
Drug Interference ◽  
Protein Levels ◽  
Novel Target ◽  
The Impact

SummaryThe tumour suppressor phosphatase and tensin homologue (PTEN), mutated or lost in many human cancers, is a major regulator of angiogenesis. However, the cellular mechanism of PTEN regulation in endothelial cells so far remains elusive. Here, we characterise the urokinase receptor (uPAR, CD87) and its tumour-derived soluble form, suPAR, as a key molecule of regulating PTEN in endothelial cells. We observed uPAR-deficient endothelial cells to express enhanced PTEN mRNA- and protein levels. Consistently, uPAR expression in endogenous negative uPAR cells, down-regulated PTEN and activated the PI3K/Akt pathway. Additionally, we found that integrin adhesion receptors act as trans-membrane signaling partners for uPAR to repress PTEN transcription in a NF-κB-dependent manner. Functional in vitro assays with endothelial cells, derived from uPAR-deficient and PTEN heterozygous crossbred mice, demonstrated the impact of uPAR- dependent PTEN regulation on cell motility and survival. In an in vivo murine angiogenesis model uPAR-deficient PTEN heterozygous animals increased the impaired angiogenic phenotype of uPAR knockout mice and were able to reverse the high invasive potential of PTEN heterozygots. Our data provide first evidence that endogenous as well as exogenous soluble uPAR down-regulated PTEN in endothelial cells to support angiogenesis. The uPAR-induced PTEN regulation might represent a novel target for drug interference, and may lead to the development of new therapeutic strategies in anti-angiogenic treatment.

scholarly journals NADP-Isocitrate Dehydrogenase fromPseudomonas nautica: Kinetic Constant Determination and Carbon Limitation Effects on the Pool of Intracellular Substrates

1998 ◽  
Vol 64 (12) ◽  
pp. 4958-4964 ◽  
Author(s):  
Sylvie O. Roy ◽  
Ted T. Packard
Keyword(s):  
Isocitrate Dehydrogenase ◽  
Protein Concentration ◽  
Marine Bacterium ◽  
Culture Medium ◽  
Kinetic Constant ◽  
Exponential Phase ◽  
Carbon Limitation ◽  
Protein Levels ◽  
Constant Determination ◽  
Michaelis Constants

ABSTRACT Variations of intracellular concentrations of isocitrate and NADP+ were measured throughout all growth phases of the marine bacterium Pseudomonas nautica. The intracellular isocitrate concentration tracked the intracellular protein concentration throughout all phases of growth. It rapidly increased in early exponential phase to a maximum and fell to nearly zero in parallel with pyruvate exhaustion in the culture medium. The intracellular NADP+ and protein concentrations increased in parallel during the exponential phase but were poorly correlated. Even after carbon exhaustion, the intracellular NADP+concentration stayed high, as did protein levels. The results demonstrated that the intracellular isocitrate concentration, but not the intracellular NADP+ concentration, was affected by the carbon availability in the culture. They also suggest that, because of its variability, isocitrate, but not NADP+, plays the larger role in the control of the respiratory CO2production rate (R CO2 ). From initial rate studies, bisubstrate Michaelis constants and the dissociation constant were determined for NADP+-specific isocitrate dehydrogenase (IDH) from P. nautica. These studies support the hypothesis that the mechanism of IDH’s activity involves the ordered addition of the substrates,d-isocitrate and NADP+. Furthermore, the results support the use of a bisubstrate enzyme kinetic equation to model R CO2 in P. nautica.

Pengaruh Lama Pengasinan Terhadap Kadar Protein Putih Telur Itik

2014 ◽  
Vol 1 (1) ◽  
pp. 36 ◽  
Author(s):  
Siti Fatimah ◽  
Muji Rahayu ◽  
Siti Aminah
Keyword(s):  
Protein Level ◽  
Egg White ◽  
Special Treatment ◽  
Egg White Protein ◽  
Protein Levels ◽  
Duck Egg ◽  
Egg Period ◽  
Graph Presentation ◽  
And Control ◽  
The Impact

Background : Egg  is one of the animal protein source, which has delicious taste, easy to digest and highly nutritious. Besides its affordable price, its supply availability is unquestionable as well. However, due to its short storability, it requires special treatment, such as preserving, to store it for long period. One way to preserve the egg is by pickling egg, which generally requires seven to ten days of marinating. During the process of marinating, there will be a visual change of egg white and yolk. Their structures  will be more solid (the occurrence of thickening process) because salinization will lead to protein denaturalization. Consequently, it has an influence as well towards the content of egg white protein of duck egg. This study is aimed to explore the impact of various time of pickling egg towards egg white protein of duck egg. Method  : The study where takes place in a laboratories, is a true experimental study for the reason that the researcher must provide intervention, hence all of potentially confounding variables are manageable. Samples that had been used in this study are duck eggs which were bought from North Brebes. This study is expected to generate data from four various time of pickling egg and control (no treatment). Since there are four samples, accordingly the number of data resulted are twenty. The resulted data will be descriptively presented in table, graph, presentation, and narration. Result  : Protein level examination within duck white egg shows changes  in protein levels that occurs in every variation of pickling egg time, where the average results of the assay of duck egg white protein is 14.94% without treatment (control), in five days of pickling time is 13.68%, in seven days of pickling time is 13.29%, in nine days of pickling time is 12.87% and eleven days of pickling time is 12.78%. Conclusion  : There is a significant impact among the period of pickling time to the protein level degradation of duck white egg. Keywords : Duck egg, period of pickling time, level protein of duck white egg.

Effects of Food Protein Levels on Haematological Parameters of Growing Japanese Quails (Coturnix Coturnix Japonica Temminck & Schlegel, 1849)

2020 ◽  
Vol 6 (6) ◽  
pp. 94-100
Author(s):  
Serge-Olivier Konan KOUASSI ◽  
◽  
Yves Bénarèce Tra DJE BI ◽  
Soualio KAMAGATE ◽  
Mathieu Nahounou BLEYERE ◽  
...  
Keyword(s):  
Crude Protein ◽  
Hematological Parameters ◽  
Coturnix Japonica ◽  
Food Protein ◽  
Coturnix Coturnix Japonica ◽  
Coturnix Coturnix ◽  
Protein Levels ◽  
Dietary Crude Protein ◽  
Japanese Quails ◽  
The Impact

The study aims to determine the impact of dietary crude protein levels on the hematological parameters of Japanese quail from growth to ovipositor. To this end, five feeds with different crude protein levels (18, 20, 22, 24 and 26%) were supplied to 700 quails of three weeks of age. These were subdivided into six batches, including three batches of females and three batches of males for each feed group. After subjecting the quails to diets containing the different protein levels, four samples were taken at the fourth, fifth, sixth and seventh week of age. The samples taken were analyzed using an SYSMEX XN 350 automated hematological analyzer. The results of this investigation indicated that non-significant differences (P > 0.05) were observed in hematological parameters in both female and male quails. This study showed that dietary crude protein levels had no impact on the health status of Japanese quails. Keywords: Japanese quails, Crude protein, Hematological parameters.

Fluoxetine Protects against Dendritic Spine Loss in Middle-aged APPswe/PSEN1dE9 Double Transgenic Alzheimer’s Disease Mice

2020 ◽  
Vol 17 (1) ◽  
pp. 93-103 ◽  
Author(s):  
Jing Ma ◽  
Yuan Gao ◽  
Wei Tang ◽  
Wei Huang ◽  
Yong Tang
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Dendritic Spines ◽  
Dendritic Spine ◽  
Early Stage ◽  
Learning Ability ◽  
Middle Aged ◽  
Protein Levels ◽  
Spine Pathology ◽  
The Impact

Background: Studies have suggested that cognitive impairment in Alzheimer’s disease (AD) is associated with dendritic spine loss, especially in the hippocampus. Fluoxetine (FLX) has been shown to improve cognition in the early stage of AD and to be associated with diminishing synapse degeneration in the hippocampus. However, little is known about whether FLX affects the pathogenesis of AD in the middle-tolate stage and whether its effects are correlated with the amelioration of hippocampal dendritic dysfunction. Previously, it has been observed that FLX improves the spatial learning ability of middleaged APP/PS1 mice. Objective: In the present study, we further characterized the impact of FLX on dendritic spines in the hippocampus of middle-aged APP/PS1 mice. Results: It has been found that the numbers of dendritic spines in dentate gyrus (DG), CA1 and CA2/3 of hippocampus were significantly increased by FLX. Meanwhile, FLX effectively attenuated hyperphosphorylation of tau at Ser396 and elevated protein levels of postsynaptic density 95 (PSD-95) and synapsin-1 (SYN-1) in the hippocampus. Conclusion: These results indicated that the enhanced learning ability observed in FLX-treated middle-aged APP/PS1 mice might be associated with remarkable mitigation of hippocampal dendritic spine pathology by FLX and suggested that FLX might be explored as a new strategy for therapy of AD in the middle-to-late stage.

scholarly journals Mechanosensation and Mechanotransduction by Lymphatic Endothelial Cells Act as Important Regulators of Lymphatic Development and Function

2021 ◽  
Vol 22 (8) ◽  
pp. 3955
Author(s):  
László Bálint ◽  
Zoltán Jakus
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Fate ◽  
Molecular Mechanisms ◽  
Critical Role ◽  
Mechanical Forces ◽  
Lymphatic Endothelial Cells ◽  
Lymphatic Development ◽  
And Function ◽  
The Impact

Our understanding of the function and development of the lymphatic system is expanding rapidly due to the identification of specific molecular markers and the availability of novel genetic approaches. In connection, it has been demonstrated that mechanical forces contribute to the endothelial cell fate commitment and play a critical role in influencing lymphatic endothelial cell shape and alignment by promoting sprouting, development, maturation of the lymphatic network, and coordinating lymphatic valve morphogenesis and the stabilization of lymphatic valves. However, the mechanosignaling and mechanotransduction pathways involved in these processes are poorly understood. Here, we provide an overview of the impact of mechanical forces on lymphatics and summarize the current understanding of the molecular mechanisms involved in the mechanosensation and mechanotransduction by lymphatic endothelial cells. We also discuss how these mechanosensitive pathways affect endothelial cell fate and regulate lymphatic development and function. A better understanding of these mechanisms may provide a deeper insight into the pathophysiology of various diseases associated with impaired lymphatic function, such as lymphedema and may eventually lead to the discovery of novel therapeutic targets for these conditions.

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