Pancreatic ductal adenocarcinoma (PDAC) is largely resistant to standard treatments leading to poor patient survival. The expression of plasma membrane calcium ATPase-4 (PMCA4) is reported to ...modulate key cancer hallmarks including cell migration, growth, and apoptotic resistance. Data-mining revealed that PMCA4 was over-expressed in pancreatic ductal adenocarcinoma (PDAC) tumors which correlated with poor patient survival. Western blot and RT-qPCR revealed that MIA PaCa-2 cells almost exclusively express PMCA4 making these a suitable cellular model of PDAC with poor patient survival. Knockdown of PMCA4 in MIA PaCa-2 cells (using siRNA) reduced cytosolic Ca2+ (Ca2+i) clearance, cell migration, and sensitized cells to apoptosis, without affecting cell growth. Knocking down PMCA4 had minimal effects on numerous metabolic parameters (as assessed using the Seahorse XF analyzer). In summary, this study provides the first evidence that PMCA4 is over-expressed in PDAC and plays a role in cell migration and apoptotic resistance in MIA PaCa-2 cells. This suggests that PMCA4 may offer an attractive novel therapeutic target in PDAC.
The microenvironment of pancreatic ductal adenocarcinoma (PDAC) is characterized by an extensive desmoplastic reaction, an alteration of the tumour stroma where activated pancreatic stellate cells ...(PSC) produce large deposits of extracellular matrix where hyaluronan (HA) stands out. The large deposits of HA have been shown to increase interstitial pressure and collapse blood vessels creating an impenetrable barrier for chemotherapy drugs. Clinical trials attempting to disrupt large deposits of HA have failed to improve chemotherapy outcomes. There are a limited number of studies that have investigated the functional role of HA in PDAC, therefore this study attempted to understand the role of exogenous and endogenous HA in two cancer hallmarks, proliferation, and migration. We used conditioned media rich in HA derived from PSCs and commercial HA to analyse the exogenous effects on PDAC proliferation and migration. This study also characterised the endogenous production of HA, the proteins involved in HA turnover and how endogenous HA affects PDAC cell proliferation and migration. Proliferation was assessed by the cell counting kit and sulforhodamine assays and migration was analysed by gap closure assays. HA production was assessed with an HAELISA-like assay and HA in PDAC cells was assessed by an immunofluorescence-like assay. The proteins involved in HA turnover were analysed by immunoblotting, cell membrane enrichment kits, immunofluorescence and siRNA knock downs. We found that PSC-conditioned media did not affect PDAC cell proliferation. Despite PSC-conditioned media increasing migration, we found that this effect was independent of HA. Exogenous HA did not affect PDAC cell proliferation. However exogenous HA had a differential effect on migration. BXPC3 cells expressing variants of HA receptors CD44 and RHAMM, increased migration with exogenous high molecular weight HA. This was abolished by blocking hyaluronidases (HYALs), HA degrading enzymes, with dextran sulphate. PDAC cells differentially produced HA. Disrupting endogenous HA by removing pericellular HA or inhibiting HA synthesis differentially decreased the migration of BXPC3 cells but did on affect PDAC cell proliferation. The combination of these two treatments had an additive effect in decreasing migration. Knocking down hyaluronan synthase 2 (HAS2) had a similar effect and greatly reduced migration in BXPC3 cells. This study provides evidence of differentially expressed HA turnover systems that can affect migration and proliferation in PDAC. Further studies investigating the role of HA in PDAC could provide insight into why the recent therapeutic attempts to target HA in the stroma have failed.
Pancreatic ductal adenocarcinoma (PDAC) is largely resistant to standard treatments leading to poor patient survival. The expression of plasma membrane calcium ATPase-4 (PMCA4) is reported to ...modulate key cancer hallmarks including cell migration, growth, and apoptotic resistance. Data-mining revealed that PMCA4 was over-expressed in pancreatic ductal adenocarcinoma (PDAC) tumors which correlated with poor patient survival. Western blot and RT-qPCR revealed that MIA PaCa-2 cells almost exclusively express PMCA4 making these a suitable cellular model of PDAC with poor patient survival. Knockdown of PMCA4 in MIA PaCa-2 cells (using siRNA) reduced cytosolic Ca
(Ca
) clearance, cell migration, and sensitized cells to apoptosis, without affecting cell growth. Knocking down PMCA4 had minimal effects on numerous metabolic parameters (as assessed using the Seahorse XF analyzer). In summary, this study provides the first evidence that PMCA4 is over-expressed in PDAC and plays a role in cell migration and apoptotic resistance in MIA PaCa-2 cells. This suggests that PMCA4 may offer an attractive novel therapeutic target in PDAC.
Introducción. La digitalización y el entorno tecnológico han permitido nuevas vías de comunicación para el feminismo que ha logrado una cobertura mediática que no hubiera sido posible sin Internet. ...El presente estudio pretende conocer cómo se estructuran los mensajes del movimiento feminista frente al reto de la convergencia a través del estudio de caso de la acción #MásMujeres implementada por la Asociación de Mujeres Cineastas y de Medios Audiovisuales (CIMA) llevada a cabo durante las ediciones de los Premios Goya desde 2018 a 2020. Metodología. La investigación recurre al análisis de contenido para identificar la cobertura mediática y el tratamiento informativo del evento, así como la difusión del contenido llevado a cabo a través de las redes sociales. Los resultados son validados a partir de una entrevista en profundidad aplicada a la responsable de la Asociación. Resultados. Los datos demuestran que la cobertura del evento ha ido disminuyendo con el paso del tiempo y que la difusión de contenido en redes sociales ha sido desigual a lo largo de los años, a pesar de las reacciones por parte de los públicos. Conclusiones y discusión. Del análisis se puede concluir que la alfombra roja responde a una herramienta oportuna de comunicación a la hora de difundir mensajes propios de la reivindicación social feminista pero que requiere de una planificación estratégica.
The space industry is currently witnessing two concurrent trends: the increased modularity and miniaturization of technologies and the deployment of constellations of distributed satellite systems. ...As a consequence of the first trend, the relevance of small satellites in line with the "cheaper and faster"philosophy is increasing. The second one opens up completely new horizons by enabling the design of architectures aimed at improving the performance, reliability, and efficiency of current and future space missions. The EU H2020 ONION project ("Operational Network of Individual Observation Nodes") has leveraged on the concept of fractionated and federated satellite systems (FFSS) to develop and design innovative mission architectures resulting in a competitive advantage for European earth observation (EO) systems. Starting from the analysis of emerging needs in the European EO market, the solutions to meet these needs are identified and characterized by exploring FFSS. In analogy with terrestrial networks, these systems envision the distribution of satellite functionalities amongst multiple cooperating spacecrafts (nodes of a network), possibly independent, and flying on different orbits. FFSS are considered by many as the future of space-based infrastructures, as they offer a pragmatic, progressive, and scalable approach to improve existing and future space missions. This paper summarizes the main results of the ONION project and the high-level design of the marine weather forecast mission for polar regions.
Prostate cancer (CaP) bone metastasis is an early event that remains inactive until later-stage progression. Reduced levels of circulating androgens, due to andropause or androgen deprivation ...therapies, alter androgen receptor (AR) coactivator expression. Coactivators shift the balance towards enhanced AR-mediated gene transcription that promotes progression to androgen-resistance. Disruptions in coregulators may represent a molecular switch that reactivates latent bone metastasis. Changes in AR-mediated transcription in androgen-sensitive LNCaP and androgen-resistant C4-2 cells were analyzed for AR coregulator recruitment in co-culture with Saos-2 and THP-1. The Saos-2 cell line derived from human osteosarcoma and THP-1 cell line representing human monocytes were used to display osteoblast and osteoclast activity. Increased AR activity in androgen-resistant C4-2 was due to increased AR expression and SRC1/TIF2 recruitment and decreased SMRT/NCoR expression. AR activity in both cell types was decreased over 90% when co-cultured with Saos-2 or THP-1 due to dissociation of AR from the SRC1/TIF2 and SMRT/NCoR coregulators complex, in a ligand-dependent and cell-type specific manner. In the absence of androgens, Saos-2 decreased while THP-1 increased proliferation of LNCaP cells. In contrast, both Saos-2 and THP-1 decreased proliferation of C4-2 in absence and presence of androgens. Global changes in gene expression from both CaP cell lines identified potential cell cycle and androgen regulated genes as mechanisms for changes in cell proliferation and AR-mediated transactivation in the context of bone marrow stroma cells.
•Decreased corepressor expression change AR in androgen-resistance prostate cancer.•Bone stroma-derived cells change AR coregulator recruitment in prostate cancer.•Bone stroma cells change cell proliferation in androgen-resistant cancer cells.•Global gene expression in CaP cells is modified by bone stroma cells in co-cultures.•Potential new multi-subunit coactivator complexes for AR in CaP bone metastasis.
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•Sonochemical-assisted synthesis Cr-TiO2@Fe3O4 was proposed as a novel method.•Cr-doped TiO2 was effective for the photodegradation of MG dye.•99% of the catalyst was recovered by a ...simple magnetic field.•The TD-DFT model predicts the by-products of the photocatalytic reaction.
Cr-TiO2 supported on Fe3O4 material was synthesized using a sonochemical method. By TEM images it was possible to observe cubic morphologies and by XRD and Raman spectroscopy the anatase phase was evaluated, showing that no subsequent thermal treatment was necessary. XPS analysis shows a decrease in the intensity of the Fe3O4 signals when the TiO2 is added to create the material discussed in this paper. Also, by this technique, it was possible to confirm the presence of Cr over the doped materials. In the photocatalysis test. 100% of color removal of the Malachite Green dye (10 mg L−1) was achieved after 300 min of reaction time under solar radiation in a cylindrical reactor with constant bubbling, and only 60% of TOC removal was reached. Using the Time Dependent – Density Functional Theory (TD-DFT) model, was possible to predict the by-products of the photocatalytic reaction of the Malachite Green dye, such as the leuco carbinol Malachite Green, 4-(dimethylamino) benzophenone and 4-4-(dimethylamino) phenyl methyl-N,N-dimethylaniline Malachite and it its compared with the experimental UV–vis spectra.
Ischemia-reperfusion (IR) injury is one of the leading causes of early graft dysfunction in liver transplantation. Techniques such as ischemic preconditioning protect the graft through the activation ...of the hypoxia-inducible factors (HIF), which are downregulated by the EGLN family of prolyl-4-hydroxylases, a potential biological target for the development of strategies based on pharmacological preconditioning. For that reason, this study aims to evaluate the effect of the EGLN inhibitor sodium (
)-2-hydroxyglutarate (
)-2HG on liver IR injury in Wistar rats.
Twenty-eight female Wistar rats were divided into the following groups: sham (SH,
= 7), non-toxicity (HGTox,
= 7, 25 mg/kg of (
)-2HG, twice per day for two days), IR (
= 7, total liver ischemia: 20 minutes, reperfusion: 60 minutes), and (
)-2HG+IR (HGIR,
= 7, 25 mg/kg of (
)-2HG, twice per day for two days, total liver ischemia as the IR group). Serum ALT, AST, LDH, ALP, glucose, and total bilirubin were assessed. The concentrations of IL-1β, IL-6, TNF, malondialdehyde, superoxide dismutase, and glutathione peroxidase were measured in liver tissue, as well as the expression of
,
, and
, determined by RT-qPCR. Sections of liver tissue were evaluated histologically, assessing the severity of necrosis, sinusoidal congestion, and cytoplasmatic vacuolization.
The administration of (
)-2HG did not cause any alteration in the assessed biochemical markers compared to SH. Preconditioning with (
)-2HG significantly ameliorated IR injury in the HGIR group, decreasing the serum activities of ALT, AST, and LDH, and the tissue concentrations of IL-1β and IL-6 compared to the IR group. IR injury decreased serum glucose compared to SH. There were no differences in the other biomarkers assessed. The treatment with (
)-2HG tended to decrease the severity of hepatocyte necrosis and sinusoidal congestion compared to the IR group. The administration of (
)-2HG did not affect the expression of
but decreased the expression of both
and
compared to the SH group, suggesting that the HIF-1 pathway is not involved in its mechanism of hepatoprotection. In conclusion, (
)-2HG showed a hepatoprotective effect, decreasing the levels of liver injury and inflammation biomarkers, without evidence of the involvement of the HIF-1 pathway. No hepatotoxic effect was observed at the tested dose.
Ischemia-reperfusion (IR) injury is the main cause of delayed graft function in solid organ transplantation. Hypoxia-inducible factors (HIFs) control the expression of genes related to ...preconditioning against IR injury. During normoxia, HIF-α subunits are marked for degradation by the egg-laying defective nine homolog (EGLN) family of prolyl-4-hydroxylases. The inhibition of EGLN stabilizes HIFs and protects against IR injury. The aim of this study was to determine whether the EGLN inhibitors sodium (
)-2-hydroxyglutarate (
)-2HG and succinic acid (SA) have a nephroprotective effect against renal IR injury in Wistar rats.
(
)-2HG was synthesized in a 22.96% yield from commercially available L-glutamic acid in a two-step methodology (diazotization/alkaline hydrolysis), and its structure was confirmed by nuclear magnetic resonance and polarimetry. SA was acquired commercially. (
)-2HG and SA were independently evaluated in male and female Wistar rats respectively after renal IR injury. Rats were divided into the following groups: sham (SH), nontoxicity (
)-2HG: 12.5 or 25 mg/kg; SA: 12.5, 25, or 50 mg/kg, IR, and compound+IR (
)-2HG: 12.5 or 25 mg/kg; SA: 12.5, 25, or 50 mg/kg; independent SH and IR groups were used for each assessed compound. Markers of kidney injury (BUN, creatinine, glucose, and uric acid) and liver function (ALT, AST, ALP, LDH, serum proteins, and albumin), proinflammatory cytokines (IL-1β, IL-6, and TNF-α), oxidative stress biomarkers (malondialdehyde and superoxide dismutase), and histological parameters (tubular necrosis, acidophilic casts, and vascular congestion) were assessed. Tissue HIF-1α was measured by ELISA and Western blot, and the expression of Hmox1 was assessed by RT-qPCR.
(
)-2HG had a dose-dependent nephroprotective effect, as evidenced by a significant reduction in the changes in the BUN, creatinine, ALP, AST, and LDH levels compared with the IR group. Tissue HIF-1α was only increased in the IR group compared to SH; however, (
)-2HG caused a significant increase in the expression of Hmox1, suggesting an early accumulation of HIF-1α in the (
)-2HG-treated groups. There were no significant effects on the other biomarkers. SA did not show a nephroprotective effect; the only changes were a decrease in creatinine level at 12.5 mg/kg and increased IR injury at 50 mg/kg. There were no effects on the other biochemical, proinflammatory, or oxidative stress biomarkers.
None of the compounds were hepatotoxic at the tested doses. (
)-2HG showed a dose-dependent nephroprotective effect at the evaluated doses, which involved an increase in the expression of Hmox1, suggesting stabilization of HIF-1α. SA did not show a nephroprotective effect but tended to increase IR injury when given at high doses.
Phthalates and bisphenols are ubiquitous environmental pollutants with the ability to perturb different systems. Specifically, they can alter the endocrine system, and this is why they are also known ...as endocrine-disrupting compounds (EDCs). Interestingly, they are related to the development and progression of breast cancer (BC), but the threshold concentrations at which they trigger that are not well established. Objectives: The aim of this study was to compare the concentration measures of parent EDCs in three groups of women (without BC, with BC, and BC survivors) from two urban populations in Mexico, to establish a possible association between EDCs and this disease. We consider the measure of the parent compounds would reflect the individual’s exposure. Methods: The levels of di-ethyl-hexyl-phthalate (DEHP), butyl-benzyl-phthalate (BBP), di-n-butyl phthalate (DBP) and di-ethyl-phthalate (DEP), bisphenol A (BPA) and bisphenol S (BPS) were determined by gas chromatograph-mass spectrometry in 102 subjects, including 37 women without any pathological disease, 46 patients with BC and 19 women survivals of BC of Mexico and Toluca City. Results: All phthalates were detected in 100% of women, two of them were significantly higher in patients with different BC subtypes in Mexico City. Differential increases were observed mainly in the serum concentration of phthalates in women with BC compared to women without disease between Mexico and Toluca City. In addition, when performing an analysis of the concentrations of phthalates by molecular type of BC, DEP and BBP were found mainly in aggressive and poorly differentiated types of BC. It should be noted that female BC survivors treated with anti-hormonal therapy showed lower levels of BBP than patients with BC. BPA and BPS were found in most samples from Mexico City. However, BPS was undetectable in women from Toluca City. Discussion: The results of our study support the hypothesis of a positive association between exposure to phthalates and BC incidence.
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