Integrin-linked kinase can facilitate syncytialization and hormonal differentiation of the human trophoblast-derived BeWo cell line

<p>Abstract</p> <p>Background</p> <p>In the fusion pathway of trophoblast differentiation, stem villous cytotrophoblast cells proliferate and daughter cells differentiate and fuse with existing syncytiotrophoblast to maintain the multi-nucleated layer. Integrin-linked kinase (ILK) is highly expressed in 1st and 2nd trimester villous cytotrophoblast cells, yet barely detectable in syncytiotrophoblast, thus we examined the potential role of ILK in aiding trophoblast fusion.</p> <p>Methods</p> <p>The temporal/spatial expression and activity of ILK were determined in BeWo cells undergoing syncytialization by immunoblot and immunofluorescence analyses. BeWo cells were also transfected with pEGFP expression vectors containing wildtype or two mutant ILK cDNA constructs. The incidence of cell fusion in transfected cells grown under syncytialization conditions was then scored by the presence or absence of E-cadherin immunostaining. Beta-hCG expression in transfected cells, a marker of syncytiotrophoblast hormonal differentiation, was also similarly assessed.</p> <p>Results</p> <p>ILK catalytic activity increased and ILK began to increasingly localize to BeWo cell nuclei during syncytialization in correlation with increased pAkt and Snail protein expression. Syncytialization was also significantly elevated (p < 0.05) in BeWo cells expressing constitutively active (ca)-ILK vs cells containing empty vector or dn-ILK. Furthermore, cytoplasmic Beta-hCG expression markedly increased (p < 0.05) in cells expressing wt- and ca-ILK.</p> <p>Conclusion</p> <p>ILK-facilitated syncytialization is dependent, at least in part, on ILK catalytic activity while hormonal differentiation appears dependent on both ILK-associated protein interactions and catalytic activity. This study demonstrates that ILK plays a novel role in BeWo syncytialization and differentiation, perhaps through an ILK-Akt-Snail pathway, and implicates ILK in the same process in villous cytotrophoblasts in vivo.</p

Soil nitrous oxide emissions from grassland: potential inhibitor effect of hippuric acid

In grassland systems, cattle and sheep urine patches are recognized as nitrous oxide (N2O) emission hot spots due to the high urinary nitrogen (N) concentrations. Hippuric acid (HA) is one of the constituents of ruminant urine that has been reported as a natural inhibitor of soil N2O emissions. The aim of this study was to examine the potential for elevated ruminant urine HA concentrations to reduce N2O emissions, in situ, on an acidic heavy clay soil under poorly drained conditions (WFPS > 85%). A randomized complete block design experiment with three replications and four treatments was conducted using the closed-static-flux chamber methodology. The four treatments were applied inside the chambers: control with no artificial urine application (C), control artificial urine (U), and enriched artificial urine containing two rates of HA (55.8 and 90 mM, U+HA1, U+HA2). Soil inorganic-N, soil dissolved organic carbon (DOC), soil pH as well as N2O and methane (CH4) fluxes were monitored over a 79-day period. Although N2O emissions were not affected by the HA enriched urine treatments, U+HA2 positively affected the retention of N as NH4+ until day 3, when the soil pH dropped to values <5. Subsequently, as a consequence of rainfall events and soil acidification, it is likely that leaching or sorption onto clay reduced the efficacy of HA, masking any treatment differential effect on N2O emissions. Moreover, CH4 fluxes as well as DOC results reflected the soil anaerobic conditions which did not favour nitrification processes. Further research is needed to determine the fate of HA into the soil which might clarify the lack of an in situ effect of this compoun

Behavioral interactions and trophic overlap between invasive signal crayfish Pacifastacus leniusculus (Decapoda, Astacidae) and native fishes in iberian rivers

The signal crayfish Pacifastacus leniusculus Dana, 1852 is a successful invasive species in the Iberian Peninsula. This is of particular conservation concern, as fish fauna is highly endemic and also threatened within this ecoregion. The aim of this study was to analyze behavioral interactions and trophic overlap between signal crayfish and native fishes in Iberian rivers (northern Spain). Video cameras were used to record fish "dominance/evasion" after spatial encounters with crayfish. Diet composition and isotopic signatures (¿13C and ¿15N) were compared to evaluate the niche overlap. Fish were dominant in 25% of the encounters with juvenile crayfish, whereas this percentage was only 4% with adult crayfish. Observations showed a high fish "evasion" response for Pyrenean stone loach Barbatula quignardi (B¿cescu-Me¿ter, 1967) (>30%). Dietary results showed a high trophic overlap between signal crayfish with the pelagic Pyrenean minnow Phoxinus bigerri Kottelat, 2007 and the benthic loach. However, the isotopic niche overlap was low, with brown trout Salmo trutta L., 1758 showing the highest area (only 0.1 ¿2). Overall, our findings suggest that interferences may occur with native species for food (i.e., benthic invertebrates). Consequently, measures should be applied to control invasive crayfish in Iberian rivers

Mechanism-Based Screen for G1/S Checkpoint Activators Identifies a Selective Activator of EIF2AK3/PERK Signalling

Human cancers often contain genetic alterations that disable G1/S checkpoint control and loss of this checkpoint is thought to critically contribute to cancer generation by permitting inappropriate proliferation and distorting fate-driven cell cycle exit. The identification of cell permeable small molecules that activate the G1/S checkpoint may therefore represent a broadly applicable and clinically effective strategy for the treatment of cancer. Here we describe the identification of several novel small molecules that trigger G1/S checkpoint activation and characterise the mechanism of action for one, CCT020312, in detail. Transcriptional profiling by cDNA microarray combined with reverse genetics revealed phosphorylation of the eukaryotic initiation factor 2-alpha (EIF2A) through the eukaryotic translation initiation factor 2-alpha kinase 3 (EIF2AK3/PERK) as the mechanism of action of this compound. While EIF2AK3/PERK activation classically follows endoplasmic reticulum (ER) stress signalling that sets off a range of different cellular responses, CCT020312 does not trigger these other cellular responses but instead selectively elicits EIF2AK3/PERK signalling. Phosphorylation of EIF2A by EIF2A kinases is a known means to block protein translation and hence restriction point transit in G1, but further supports apoptosis in specific contexts. Significantly, EIF2AK3/PERK signalling has previously been linked to the resistance of cancer cells to multiple anticancer chemotherapeutic agents, including drugs that target the ubiquitin/proteasome pathway and taxanes. Consistent with such findings CCT020312 sensitizes cancer cells with defective taxane-induced EIF2A phosphorylation to paclitaxel treatment. Our work therefore identifies CCT020312 as a novel small molecule chemical tool for the selective activation of EIF2A-mediated translation control with utility for proof-of-concept applications in EIF2A-centered therapeutic approaches, and as a chemical starting point for pathway selective agent development. We demonstrate that consistent with its mode of action CCT020312 is capable of delivering potent, and EIF2AK3 selective, proliferation control and can act as a sensitizer to chemotherapy-associated stresses as elicited by taxanes

EDUCORE project: a clinical trial, randomised by clusters, to assess the effect of a visual learning method on blood pressure control in the primary healthcare setting

<p>Abstract</p> <p>Background</p> <p>High blood pressure (HBP) is a major risk factor for cardiovascular disease (CVD). European hypertension and cardiology societies as well as expert committees on CVD prevention recommend stratifying cardiovascular risk using the SCORE method, the modification of lifestyles to prevent CVD, and achieving good control over risk factors. The EDUCORE (Education and Coronary Risk Evaluation) project aims to determine whether the use of a cardiovascular risk visual learning method - the EDUCORE method - is more effective than normal clinical practice in improving the control of blood pressure within one year in patients with poorly controlled hypertension but no background of CVD;</p> <p>Methods/Design</p> <p>This work describes a protocol for a clinical trial, randomised by clusters and involving 22 primary healthcare clinics, to test the effectiveness of the EDUCORE method. The number of patients required was 736, all between 40 and 65 years of age (n = 368 in the EDUCORE and control groups), all of whom had been diagnosed with HBP at least one year ago, and all of whom had poorly controlled hypertension (systolic blood pressure ≥ 140 mmHg and/or diastolic ≥ 90 mmHg). All personnel taking part were explained the trial and trained in its methodology. The EDUCORE method contemplates the visualisation of low risk SCORE scores using images embodying different stages of a high risk action, plus the receipt of a pamphlet explaining how to better maintain cardiac health. The main outcome variable was the control of blood pressure; secondary outcome variables included the SCORE score, therapeutic compliance, quality of life, and total cholesterol level. All outcome variables were measured at the beginning of the experimental period and again at 6 and 12 months. Information on sex, age, educational level, physical activity, body mass index, consumption of medications, change of treatment and blood analysis results was also recorded;</p> <p>Discussion</p> <p>The EDUCORE method could provide a simple, inexpensive means of improving blood pressure control, and perhaps other health problems, in the primary healthcare setting;</p> <p>Trial registration</p> <p>The trial was registered with ClinicalTrials.gov, number NCT01155973 [<url>http://ClinicalTrials.gov</url>].</p

EnviroPlaNet Project: A systematic monitoring of atmospheric deposition of microplastics in Spain

SETAC EUROPE 33RD ANNUAL MEETING 30 APRIL-4 MAY 2023 | DUBLIN, IRELANDPlastic pollution is a global concern. The world's production continues to increase yearly, reaching more than 367 million tons in 2020 [1]. Part of these plastics are directly manufactured as microplastics (MPs) while another fraction becomes MPs as a result of the degradation processes of larger plastics into microsized particles. In this context, MPs have already been found in the hydrosphere, biosphere, cryosphere, lithosphere and, recently, atmosphere. However, the presence of MPs in the atmosphere has been poorly studied. The objective of this work was the systematic monitoring of the deposition of MPs in areas of Spain with different characteristics (population, economic activities, and climate) using a common method for sampling, quantification, and identification during the four seasons of a one-year sampling campaign with the purpose of clarifying the role of the atmosphere in the fate and dispersion of MPs. This was possible thanks to the collaboration of several Spanish research groups in the project Thematic Network of Micro- and Nanoplastics in the EnvironmentPeer reviewe

Lipid (per) oxidation in mitochondria:an emerging target in the ageing process?

Lipids are essential for physiological processes such as maintaining membrane integrity, providing a source of energy and acting as signalling molecules to control processes including cell proliferation, metabolism, inflammation and apoptosis. Disruption of lipid homeostasis can promote pathological changes that contribute towards biological ageing and age-related diseases. Several age-related diseases have been associated with altered lipid metabolism and an elevation in highly damaging lipid peroxidation products; the latter has been ascribed, at least in part, to mitochondrial dysfunction and elevated ROS formation. In addition, senescent cells, which are known to contribute significantly to age-related pathologies, are also associated with impaired mitochondrial function and changes in lipid metabolism. Therapeutic targeting of dysfunctional mitochondrial and pathological lipid metabolism is an emerging strategy for alleviating their negative impact during ageing and the progression to age-related diseases. Such therapies could include the use of drugs that prevent mitochondrial uncoupling, inhibit inflammatory lipid synthesis, modulate lipid transport or storage, reduce mitochondrial oxidative stress and eliminate senescent cells from tissues. In this review, we provide an overview of lipid structure and function, with emphasis on mitochondrial lipids and their potential for therapeutic targeting during ageing and age-related disease

A nationwide monitoring of atmospheric microplastic deposition

8 Pág.Plastic production continues to increase every year, yet it is widely acknowledged that a significant portion of this material ends up in ecosystems as microplastics (MPs). Among all the environmental compartments affected by MPs, the atmosphere remains the least well-known. Here, we conducted a one-year simultaneous monitoring of atmospheric MPs deposition in ten urban areas, each with different population sizes, economic activities, and climates. The objective was to assess the role of the atmosphere in the fate of MPs by conducting a nationwide quantification of atmospheric MP deposition. To achieve this, we deployed collectors in ten different urban areas across continental Spain and the Canary Islands. We implemented a systematic sampling methodology with rigorous quality control/quality assurance, along with particle-oriented identification and quantification of anthropogenic particle deposition, which included MPs and industrially processed natural fibres. Among the sampled MPs, polyester fibres were the most abundant, followed by acrylic polymers, polypropylene, and alkyd resins. Their equivalent sizes ranged from 22 μm to 398 μm, with a median value of 71 μm. The particle size distribution of MPs showed fewer large particles than expected from a three-dimensional fractal fragmentation pattern, which was attributed to the higher mobility of small particles, especially fibres. The atmospheric deposition rate of MPs ranged from 5.6 to 78.6 MPs m-2 day-1, with the higher values observed in densely populated areas such as Barcelona and Madrid. Additionally, we detected natural polymers, mostly cellulosic fibres with evidence of industrial processing, with a deposition rate ranging from 6.4 to 58.6 particles m-2 day-1. There was a positive correlation was found between the population of the study area and the median of atmospheric MP deposition, supporting the hypothesis that urban areas act as sources of atmospheric MPs. Our study presents a systematic methodology for monitoring atmospheric MP deposition.The authors acknowledge the support provided by Spanish Network of Plastics in the Environment, EnviroPlaNet (www.enviroplanet.net) and the financial support provided by the Spanish Government, Ministerio de Ciencia e Innovación, grants PID2020-113769RB-C21/C22. The authors would like to thank the Interdepartmental Investigation Research Service of the Universidad Autónoma de Madrid (SIDI-UAM and Segainvex) for the use of their infrastructures and their technical support. J.G.S. thanks ACIISI for the contract from the Viera y Clavijo program at the University of La Laguna (85 % co-financed by the European Social Fund).Peer reviewe

Molecular mechanisms of cell death: recommendations of the Nomenclature Committee on Cell Death 2018.

Over the past decade, the Nomenclature Committee on Cell Death (NCCD) has formulated guidelines for the definition and interpretation of cell death from morphological, biochemical, and functional perspectives. Since the field continues to expand and novel mechanisms that orchestrate multiple cell death pathways are unveiled, we propose an updated classification of cell death subroutines focusing on mechanistic and essential (as opposed to correlative and dispensable) aspects of the process. As we provide molecularly oriented definitions of terms including intrinsic apoptosis, extrinsic apoptosis, mitochondrial permeability transition (MPT)-driven necrosis, necroptosis, ferroptosis, pyroptosis, parthanatos, entotic cell death, NETotic cell death, lysosome-dependent cell death, autophagy-dependent cell death, immunogenic cell death, cellular senescence, and mitotic catastrophe, we discuss the utility of neologisms that refer to highly specialized instances of these processes. The mission of the NCCD is to provide a widely accepted nomenclature on cell death in support of the continued development of the field