Two major mechanisms of intracellular protein degradation, autophagy and the ubiquitin-proteasome pathway, operate in mammalian cells. PTEN, which is frequently mutated in glioblastomas, is a tumor ...suppressor gene that encodes a dual specificity phosphatase that antagonizes the phosphatidylinositol 3-kinase class I/AKT/mTOR pathway, which is a key regulator of autophagy. Here, we investigated in U87MG human glioma cells the role of PTEN in the regulation of autophagy and the ubiquitin-proteasome pathway, because both are functionally linked and are relevant in cancer progression. Since U87MG glioma cells lack a functional PTEN, we used stable clones that express, under the control of a tetracycline-inducible system (Tet-on), wild-type PTEN and two of its mutants, G129E-PTEN and C124S-PTEN, which, respectively, lack the lipid phosphatase activity only and both the lipid and the protein phosphatase activities of this protein. Expression of PTEN in U87MG glioma cells decreased proteasome activity and also reduced protein ubiquitination. On the contrary, expression of PTEN increased the autophagic flux and the lysosomal mass. Interestingly, and although PTEN negatively regulates the phosphatidylinositol 3-kinase class I/AKT/mTOR signaling pathway by its lipid phosphatase activity, both effects in U87MG cells were independent of this activity. These results suggest a new mTOR-independent signaling pathway by which PTEN can regulate in opposite directions the main mechanisms of intracellular protein degradation.
GTPBP3 is an evolutionary conserved protein presumably involved in mitochondrial tRNA (mt-tRNA) modification. In humans, GTPBP3 mutations cause hypertrophic cardiomyopathy with lactic acidosis, and ...have been associated with a defect in mitochondrial translation, yet the pathomechanism remains unclear. Here we use a GTPBP3 stable-silencing model (shGTPBP3 cells) for a further characterization of the phenotype conferred by the GTPBP3 defect. We experimentally show for the first time that GTPBP3 depletion is associated with an mt-tRNA hypomodification status, as mt-tRNAs from shGTPBP3 cells were more sensitive to digestion by angiogenin than tRNAs from control cells. Despite the effect of stable silencing of GTPBP3 on global mitochondrial translation being rather mild, the steady-state levels and activity of Complex I, and cellular ATP levels were 50% of those found in the controls. Notably, the ATPase activity of Complex V increased by about 40% in GTPBP3 depleted cells suggesting that mitochondria consume ATP to maintain the membrane potential. Moreover, shGTPBP3 cells exhibited enhanced antioxidant capacity and a nearly 2-fold increase in the uncoupling protein UCP2 levels. Our data indicate that stable silencing of GTPBP3 triggers an AMPK-dependent retrograde signaling pathway that down-regulates the expression of the NDUFAF3 and NDUFAF4 Complex I assembly factors and the mitochondrial pyruvate carrier (MPC), while up-regulating the expression of UCP2. We also found that genes involved in glycolysis and oxidation of fatty acids are up-regulated. These data are compatible with a model in which high UCP2 levels, together with a reduction in pyruvate transport due to the down-regulation of MPC, promote a shift from pyruvate to fatty acid oxidation, and to an uncoupling of glycolysis and oxidative phosphorylation. These metabolic alterations, and the low ATP levels, may negatively affect heart function.
Patients with non-ST-elevation acute coronary syndrome (NSTE-ACS) are often managed conservatively. Clinical practice guidelines recommend treating these patients with the same pharmacological drugs ...as those who receive invasive treatment. We analyze the use of new antiplatelet drugs (NADs) and other recommended treatments in people discharged following an NSTE-ACS according to the treatment strategy used, comparing the medium-term prognosis between groups.
Prospective observational multicenter registry study in 1717 patients discharged from hospital following an ACS; 1143 patients had experienced an NSTE-ACS. We analyzed groups receiving the following treatment: No cardiac catheterization (NO CATH): n = 134; 11.7%; Cardiac catheterization without revascularization (CATH-NO REVASC): n = 256; 22.4%; percutaneous coronary intervention (PCI): n = 629; 55.0%; and coronary artery bypass graft (CABG): n = 124; 10.8%. We assessed major adverse cardiovascular events (MACE), all-cause mortality, and hemorrhagic complications at one year.
NO CATH was the oldest, had the most comorbidities, and was at the highest risk for ischemic and hemorrhagic events. Few patients who were not revascularized with PCI received NADs (NO CATH: 3.7%; CATH-NO REVASC: 10.6%; PCI: 43.2%; CABG: 3.2%; p<0.001). Non-revascularized patients also received fewer beta-blockers, angiotensin-converting enzyme (ACE) inhibitors, angiotensin II receptor blockers (ARB), and statins (p<0.001). At one year, MACE incidence in NO CATH group was three times that of the other groups (30.1%, p<0.001), and all-cause mortality was also much higher (26.3%, p<0.001). There were no significant differences in hemorrhagic events. Belonging to NO CATH group was an independent predictor for MACE at one year in the multivariate analysis (HR 2.72, 95% CI 1.29-5.73; p = 0.008).
Despite current invasive management of NSTE-ACS, patients not receiving catheterization are at very high risk for under treatment with recommended drugs, including NADs. Their medium-term prognosis is poor, with high mortality. Patients treated with PCI receive better pharmacological management, with high use of NADs.
Information regarding inborn error of immunity (IEI) as a risk factor for severe COVID-19 is scarce. We aimed to determine if paediatric patients with moderate/severe IEI got COVID-19 at the same ...level as the general population, and to describe COVID-19 expression.
We included patients with moderate/severe IEI aged 0–21 years old: cross-sectional study (June2020) to determine the prevalence of COVID-19; prospective study (January2020-January2021) including IEI patients with COVID-19. Assays used: nasopharyngeal swab SARS-CoV-2 PCR and SARS-CoV-2-specific immunoglobulins.
Seven from sixty-five patients tested positive (prevalence: 10.7% (7%–13%)) after the first SARS-COV-2 wave and 13/15 patients diagnosed with COVID-19 had an asymptomatic/mild course.
In our area, prevalence of COVID-19 in moderate/severe IEI paediatric patients after the first wave was slightly higher than in the general population. The majority of patients presented a benign course, suggesting a possible protective factor related with age despite IEI.
There is an increasing trend toward understanding the impact of non-
yeasts on the winemaking process. Although
is the predominant species at the end of fermentation, it has been recognized that the ...presence of non-
species during alcoholic fermentation can produce an improvement in the quality and complexity of the final wines. A previous work was developed for selecting the best combinations between
and five non-
(
, and
) native yeast strains from D.O. "Vinos de Madrid" at the laboratory scale. The best inoculation strategies between
and non-
strains were chosen to analyze, by real-time quantitative PCR (qPCR) combined with the use of specific primers, the dynamics of inoculated populations throughout the fermentation process at the pilot scale using the Malvar white grape variety. The efficiency of the qPCR system was verified independently of the samples matrix, founding the inoculated yeast species throughout alcoholic fermentation. Finally, we can validate the positive effect of selected co-cultures in the Malvar wine quality, highlighting the sequential cultures of
CLI 918/
CLI 889 and
CLI 920/
CLI 889 and, mixed and sequential cultures of
9-6C combined with
CLI 889.
This study aims to determine whether using hsTn results in medium-term prognostic differences in patients with unstable angina and NSTEMI. This multicenter, prospective registry study included ...consecutive patients who underwent hsTn assays and were discharged with a diagnosis of NSTEACS. Medium-term incidence of MACE was similar in patients with unstable angina and NSTEMI (p=0.79), but cardiovascular (p=0.012) and all-cause mortality (p=0.002) in NSTEMI patients was over twice that of patients with UA.
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The introduction of high-sensitivity troponin (hsTn) assays has reduced the diagnosis of unstable angina (UA) in favor of non-ST elevation myocardial infarction (NSTEMI) in the context of non-ST elevation acute coronary syndrome (NSTEACS). It is unclear whether the detection of these hsTn levels affects the prognosis and therefore whether a different therapeutic approach is warranted. This study aims to determine whether using hsTn results in medium-term prognostic differences in patients with UA and NSTEMI.
This multicenter, prospective registry study included consecutive patients who underwent hsTn assays and were discharged with a diagnosis of NSTEACS. Patients were followed for two years. Outcomes were the occurrence of major adverse cardiovascular events (MACE: cardiovascular death, non-fatal myocardial infarction, and non-fatal ischemic stroke), major bleeding, and all-cause mortality.
Patients with UA and NSTEMI did not show differences in terms of the invasive interventions received, the coronary artery disease diagnosed, the type of revascularization performed, or the proportion presenting MACE (UA 18.1% vs. NSTEMI 18.9%; p = 0.79). However, patients with NSTEMI had higher cardiovascular mortality at two years (UA 4% vs. NSTEMI 9.2%; p = 0.012), as well as, all-cause mortality (UA vs. 7.9% vs. NSTEMI 16.4%; p = 0.002).
Medium-term incidence of MACE was similar in patients with UA and NSTEMI, but cardiovascular and all-cause mortality in NSTEMI patients was over twice that of patients with UA.
The DOLAM trial revealed that switching from triple antiretroviral therapy (three-drug regimen; 3DR) to dolutegravir plus lamivudine (two-drug regimen; 2DR) was virologically non-inferior to ...continuing 3DR after 48 weeks of follow-up. Weight increased with 2DR relative to 3DR but it did not impact on metabolic parameters.
Multiomics plasma profile was performed to gain further insight into whether this therapy switch might affect specific biological pathways. DOLAM (EudraCT 201500027435) is a Phase 4, randomized, open-label, non-inferiority trial in which virologically suppressed persons with HIV treated with 3DR were assigned (1:1) to switch to 2DR or to continue 3DR for 48 weeks. Untargeted proteomics, metabolomics and lipidomics analyses were performed at baseline and at 48 weeks. Univariate and multivariate analyses were performed to identify changes in key molecules between both therapy arms.
Switching from 3DR to 2DR showed a multiomic impact on circulating plasma concentration of N-acetylmuramoyl-L-alanine amidase (Q96PD5), insulin-like growth factor-binding protein 3 (A6XND0), alanine and triglyceride (TG) (48:0). Correlation analyses identified an association among the up-regulation of these four molecules in persons treated with 2DR.
Untargeted multiomics profiling studies identified molecular changes potentially associated with inflammation immune pathways, and with lipid and glucose metabolism. Although these changes could be associated with potential metabolic or cardiovascular consequences, their clinical significance remains uncertain. Further work is needed to confirm these findings and to assess their long-term clinical consequences.
As more restrictions on tobacco marketing communication are implemented, tobacco marketing has persisted through smoking in films. Our aims were to assess changes in tobacco imagery exposure in ...Spanish top-grossing films before and after the banning of tobacco advertising in Spain, and to determine whether the depiction of smoking characters has changed over the years.
A repeated cross-sectional study measured the tobacco content in the 10 Spanish top-grossing films in 2005, 2010 and 2015 (n=30) before and after a complete tobacco advertising ban. We conducted a descriptive and regression analysis of changes in tobacco impressions by year.
The 30 films contained 1378 tobacco occurrences (90.2% positive for tobacco) with a median length of eight seconds onscreen. Total tobacco occurrences deemed positive for tobacco interests significantly increased in 2010 and 2015 compared to 2005. However, we observed decreased odds of tobacco brands appearances (OR=0.25; p<0.001) in 2010 and of implied tobacco use (OR=0.44; p=0.002), and tobacco brands appearances (OR=0.36; p<0.001) in 2015 compared to 2005. There was a change of pattern in the type of role smokers played from a leading role to a supporting one (p<0.001). The population reach of positive for tobacco occurrence in Spanish top-grossing films decreased from 15.9 (95% CI: 15.86-15.86) per 1000 spectators in 2005 to 0.8 (95% CI: 0.82-0.82) in 2015.
The implementation of a ban on complete tobacco product advertising was followed by a decrease in tobacco incidents across top-grossing Spanish films. Yet, exposure to smoking in films is still unacceptably high.
Microbial Desalination Cell (MDC) represents an innovative technology which accomplishes simultaneous desalination and wastewater treatment without external energy input. MDC technology could be ...employed to provide freshwater with low-energy input, for example, in remote areas where organic wastes (i.e., urban or industrial) are available. In addition, MDC technology has been proposed as pre-treatment in conventional reverse osmosis plants, with the aim of saving energy and avoiding greenhouse gases related to conventional desalination processes. The use of oxygen reduction (i.e. O2 + 2H2O + 4e− → 4 OH−, E0′ = 0.815 V, pH = 7) was usually implemented as cathodic reaction in most of the MDCs reported in literature, whereas other strategies based on liquid catholytes have been also proposed, for example, ferro-ferricyanide redox couple (i.e. Fe(CN)63- + 1e− → Fe(CN)64-, E0 = 0.37 V). As the MDC designs in the literature and operation modes (i.e., batch, continuous, semi-continuous, etc.) are quite different, the available MDC studies are not directly comparable. For this reason, the main objective of this work was to have a proper comparison of two similar MDCs operating with two different catholyte strategies, and compare performance and desalination efficiencies. In this sense, this study compares the desalination performance of two laboratory-scale MDCs located in two different locations for brackish water and sea water using two different strategies. The first strategy consisted of an air cathode for efficient oxygen reduction, while the second strategy was based on a liquid catholyte with Fe3+/Fe2+ solution (i.e., ferro-ferricyanide complex). Both strategies achieved desalination efficiency above 90% for brackish water. Nominal desalination rates (NDR) were in the range of 0.17–0.14 L·m−2·h−1 for brackish and seawater with air diffusion cathode MDC, respectively, and 1.5–0.7 L·m−2·h−1 when using ferro-ferricyanide redox MDC. Organic matter present in wastewater was effectively removed at 0.9 and 1.1 kg COD·m−3·day−1 using the air diffusion cathode MDC for brackish and sea water, respectively, and 7.1 and 19.7 kg COD·m−3·day−1 with a ferro-ferricyanide redox MDC. Both approaches used a laboratory MDC prototype without any energy supply (excluding pumping energy). Pros and cons of both strategies are discussed for subsequent upscaling of MDC technology.
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