The members of the superfamily of Transient Receptor Potential (TRP) ion channels are physiologically important molecules that have been studied for many years and are still being intensively ...researched. Among the vanilloid TRP subfamily, the TRPV4 ion channel is an interesting protein due to its involvement in several essential physiological processes and in the development of various diseases. As in other proteins, changes in its function that lead to the development of pathological states, have been closely associated with modification of its regulation by different molecules, but also by the appearance of mutations which affect the structure and gating of the channel. In the last few years, some structures for the TRPV4 channel have been solved. Due to the importance of this protein in physiology, here we discuss the recent progress in determining the structure of the TRPV4 channel, which has been achieved in three species of animals (
,
, and
), highlighting conserved features as well as key differences among them and emphasizing the binding sites for some ligands that play crucial roles in its regulation.
Wastewater treatment plants (WWTPs), linking human fecal residues and the environment, are considered as hotspots for the spread of antimicrobial resistance (AMR). In order to evaluate the role of ...WWTPs and underlying operational parameters for the removal of AMR, the presence and removal efficiency of a selected set of 6 antimicrobial resistance genes (ARGs) and 2 mobile genetic elements (MGEs) was evaluated by means of qPCR in influent and effluent samples from 62 Dutch WWTPs. The role of possible factors impacting the concentrations of ARGs and MGEs in the influent and their removal was identified through statistical analysis. ARGs and the class I integron-integrase gene (intI1) were, on average, removed to a similar extent (1.76 log reduction) or better (+0.30–1.90 logs) than the total bacteria (measured as 16S rRNA gene). In contrast, broad-host-range plasmids (IncP-1) had a significantly increased (p < 0.001) relative abundance after treatment. The presence of healthcare institutions in the area served did only slightly increase the concentrations of ARGs or MGEs in influent. From the extended panel of operational parameters, rainfall, increasing the hydraulic load of the plant, most significantly (p < 0.05) affected the treatment efficiency by decreasing it on average −0.38 logs per time the flow exceeded the average daily flow. Our results suggest that overall, WWTP treatments do not favor the proliferation of the assessed resistance genes but might increase the relative abundance of broad-host-range plasmids of the IncP-1 type.
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•Sulfonamide (sul1) and macrolide (ermB) ARGs are predominant ARGs in the influent.•Healthcare institutions in the catchment didn't increase the influent's ARGs loads.•WWTP treatment reduced ARGs by 1.8–2.7 logs.•Relative abundance of IncP-1 plasmids was increased (p < 0.001) after WWTP treatment.•Rainfall increasing the WWTP hydraulic load significantly reduced the removal of ARGs.
The mesenchymal phenotype of glioblastoma multiforme (GBM), the most frequent and malignant brain tumor, is associated with the worst prognosis. The epithelial-mesenchymal transition (EMT) is a cell ...plasticity mechanism involved in GBM malignancy. In this study, we determined 17β-estradiol (E2)-induced EMT by changes in cell morphology, expression of EMT markers, and cell migration and invasion assays in human GBM-derived cell lines. E2 (10 nM) modified the shape and size of GBM cells due to a reorganization of actin filaments. We evaluated EMT markers expression by RT-qPCR, Western blot, and immunofluorescence.We found that E2 upregulated the expression of the mesenchymal markers, vimentin, and N-cadherin. Scratch and transwell assays showed that E2 increased migration and invasion of GBM cells. The estrogen receptor-α (ER-α)-selective agonist 4,4',4''-(4-propyl-1H-pyrazole-1,3,5-triyl)trisphenol (PPT, 10 nM) affected similarly to E2 in terms of the expression of EMT markers and cell migration, and the treatment with the ER-α antagonist methyl-piperidino-pyrazole (MPP, 1 μM) blocked E2 and PPT effects. ER-β-selective agonist diarylpropionitrile (DNP, 10 nM) and antagonist 4-2-phenyl-5,7-bis(trifluoromethyl)pyrazole1,5-apyrimidin-3-ylphenol (PHTPP, 1 μM) showed no effects on EMT marker expression. These data suggest that E2 induces EMT activation through ER-α in human GBM-derived cells.
Glioblastomas (GBM) are the most frequent and aggressive brain tumors. In these malignancies, progesterone (P4) promotes proliferation, migration, and invasion. The P4 metabolite allopregnanolone ...(3α-THP) similarly promotes cell proliferation in the U87 human GBM cell line. Here, we evaluated global changes in gene expression of U87 cells treated with 3α-THP, P4, and the 5α-reductase inhibitor, finasteride (F). 3α-THP modified the expression of 137 genes, while F changed 90. Besides, both steroids regulated the expression of 69 genes. After performing an over-representation analysis of gene ontology terms, we selected 10 genes whose products are cytoskeleton components, transcription factors, and proteins involved in the maintenance of DNA stability and replication to validate their expression changes by RT-qPCR. 3α-THP up-regulated six genes, two of them were also up-regulated by F. Two genes were up-regulated by P4 alone, however, such an effect was blocked by F when cells were treated with both steroids. The remaining genes were regulated by the combined treatments of 3α-THP + F or P4 + F. An in-silico analysis revealed that promoters of the six up-regulated genes by 3α-THP possess cyclic adenosine monophosphate (cAMP) responsive elements along with CCAAT/Enhancer binding protein alpha (CEBPα) binding sites. These findings suggest that P4 and 3α-THP regulate different sets of genes that participate in the growth of GBMs.
Epithelial-mesenchymal transition (EMT) is an essential mechanism contributing to glioblastoma multiforme (GBM) progression, the most common and malignant brain tumor. EMT is induced by signaling ...pathways that crosstalk and regulate an intricate regulatory network of transcription factors. It has been shown that downstream components of 17β-estradiol (E2) and transforming growth factor β (TGF-β) signaling pathways crosstalk in estrogen-sensitive tumors. However, little is known about the interaction between the E2 and TGF-β signaling components in brain tumors. We have investigated the relationship between E2 and TGF-β signaling pathways and their effects on EMT induction in human GBM-derived cells. Here, we showed that E2 and TGF-β negatively regulated the expression of estrogen receptor α (ER-α) and Smad2/3. TGF-β induced Smad2 phosphorylation and its subsequent nuclear translocation, which E2 inhibited. Both TGF-β and E2 induced cellular processes related to EMT, such as morphological changes, actin filament reorganization, and mesenchymal markers (N-cadherin and vimentin) expression. Interestingly, we found that the co-treatment of E2 and TGF-β blocked EMT activation. Our results suggest that E2 and TGF-β signaling pathways interact through ER-α and Smad2/3 mediators in cells derived from human GBM and inhibit EMT activation induced by both factors alone.
•5α-DHP promotes proliferation of glioblastoma cells similarly to progesterone (P4)•5α-DHP induction of proliferation is blocked by P4 receptor (PR) antagonist, RU486.•5α-DHP promotes migration of ...glioblastoma cells through the PR.
Glioblastomas (GBMs) are the most common and deadliest intracranial tumors. Steroid hormones, such as progesterone (P4), at physiological concentrations, promote proliferation, and migration of human GBM cells in vivo and in vitro. Neuronal and glial cells, but also GBMs, metabolize P4 and synthesize different active metabolites such as 5α-dihydroprogesterone (5α-DHP). However, their contribution to GBM malignancy remains unknown. Here, we determined the 5α-DHP effects on the number of cells, proliferation, and migration of the U87 and U251 human GBM-derived cell lines. Of the tested concentrations (1 nM-1 µM), 5α-DHP 10 nM significantly increased the number of U87 and U251 cells from day 2 of treatment, and proliferation (at day 3) in a similar manner as P4 (10 nM). The treatment with the progesterone receptor (PR) antagonist RU486 (mifepristone), blocked the effects of 5α-DHP on the number of cells and proliferation. Besides, in U251 and LN229 GBM cells, 5α-DHP promoted cell migration (from 12 to 24 h). We also determined that GBM cells expressed the 3α-hydroxysteroid oxidoreductases (3α-HSOR), which reversibly reduce 5α-DHP to allopregnanolone (3α-THP). These data indicate that 5α-DHP induces proliferation and migration of human GBM through the activation of PR.
Glioblastomas (GBM) are the most frequent and aggressive brain tumors due to their recurrence and resistance to current therapies. These characteristics are associated with the presence of glioma ...stem cells (GSCs), mainly identified by the detection of the membrane antigens CD133 and CD15. The main source of GSCs has been biopsies of tumors. However, alternatives are sought from cell lines because more homogeneous populations can be obtained with high yields. This chapter describes a method for the enrichment and characterization of GSCs from cell lines derived from human GBM by selective culture with serum-free neural stem cell medium and growth factors. The technique offers alternatives for the enrichment and characterization of GSCs, that could contribute to a better understanding of the biology of GBMs.
AbstractBackground and aims. Secreted frizzled-related protein 5 (SFRP5) was recently described as a new adipokine protective for hepatic steatosis and other obesity-related complications in the ...mouse model. To date, SFRP5 expression in non-alcoholic fatty liver disease (NAFLD) has not been fully assessed in humans. We measured circulating SFRP5 levels and its expression in liver and adipose tissue, and evaluated its association with NAFLD in morbidly obese women. Material and methods. Fifty-four morbidly obese women undergoing bariatric surgery were included in the study. Liver biopsies were used for histology and hepatic triglyceride content quantification. Circulating SFRP5 levels were measured through enzyme-linked immunoabsorbent assay, and SFRP5 expression was performed in hepatic and adipose tissue (subcutaneous and visceral). Results. Although circulating SFRP5 levels showed a tendency to decrease with NAFLD progression, no significant differences were observed among non-alcoholic steatosis, steatohepatitis, and control subjects. Hepatic SFRP5 expression showed a negative correlation with hepatic triglyceride content (r = -0.349, P = 0.016 for mRNA and r = -0.291, P = 0.040 for SRFP5 protein) and ALT serum levels (r = -0.437, P = 0.001 for SRFP5 protein). In addition, hepatic SFRP5 protein levels were significantly lower in NASH than in control subjects (P = 0.006). Conclusion. This is the first study reporting an association of hepatic SFRP5 expression with NAFLD in humans.
This work presents several interactive learning materials developed by the Group of Educational Innovation Manufacturing Engineering of the University of Las Palmas de Gran Canaria. This work ...presents the methodology followed, the main results achieved, the first classroom experiences, their evolution, new materials that are being developed, and the conclusions obtained from this line of work.
Background: Superficial venous thrombosis (SVT) is a common clinical condition caused by inflammation and the presence of a thrombus inside a superficial vein. It has traditionally been considered a ...benign and banal disorder, although it can progress or can be associated with thromboembolic disease of deep territories in up to 20%, asymptomatic or symptomatic pulmonary embolism (PE), especially if it affects the main trunk of the internal saphenous vein. The impact of deep vein thrombosis on the quality of life and its sequelae have long been described in the literature; however, they have not been studied in superficial vein thrombosis. Objectives: We aimed to evaluate the risk factors, management, and complications of SVT and its impact on the quality of life of our patients. Methods: Observational, prospective, single-center study to evaluate the management of SVT. The ultrasound (US) was performed initially on symptomatic patients, during treatment with low-molecular-weight heparin (LMWH), at a follow-up, and at the end of 45 days of treatment. A quality-of-life questionnaire was administered to determine the risk factors, management, and complications of SVT at the moment of diagnosis and at the end of treatment. We included patients referred from the emergency department to a monographic consultation for thromboembolic disease, over 18 years of age with a diagnosis of acute SVT symptomatic, without contraindication to initiate anticoagulation. Results: In total, 63 patients were evaluated between October 2020 and April 2022. The mean age was 65.8 years (SD 13.5), of which 35 were women (55.6%), 39 presented cardiovascular risk factors (61.9%), 25 had a history of previous personal venous thromboembolism (VTE) (39.7%), and 10 had obesity (15.9%), 47 had chronic venous insufficiency or varicose veins (74.9%). During follow-up with ultrasound, 39.7% had partial revascularization, and at discharge, 63.5% had permeabilized the thrombosis against 19% who had residual thrombosis or progression of thrombosis. There was a positive correlation between mobility parameters and improvement in the performance of daily activities (rho = 0.35; p = 0.012) and with improvement in pain/discomfort (rho = 0.37; p = 0.007). An improvement in pain parameters was statistically significantly related to a global assessment health perception (rho = 0.48; p < 0.001). Anxiety and depression parameters were related to a global assessment health perception (rho = 0.462; p = 0.001) and to an overall improvement at 12 months (rho = 0.45; p = 0.001). CONCLUSIONS: Superficial venous thrombosis (SVT) is a highly prevalent disease, which is traditionally considered banal and has good evolution, with heterogeneous management in clinical practice and limited information on patient selection for therapies, current treatment routes, and drug use, as well as outcomes. In recent years, the importance of this entity has become evident due to its frequency in clinical practice, its risk of complications, and the impact it has on the quality of life. This study’s results emphasize the importance of the diagnosis, treatment, and follow-up of superficial venous thrombosis.
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