Next-Generation Sequencing has recently been introduced to efficiently and simultaneously detect genetic variations in acute myeloid leukemia. However, its implementation in the clinical routine ...raises new challenges focused on the diversity of assays and variant reporting criteria. To overcome this challenge, the PETHEMA group established a nationwide network of reference laboratories aimed to deliver molecular results in the clinics. We report the technical cross-validation results for next-generation sequencing panel genes during the standardization process and the clinical validation in 823 samples of 751 patients with newly diagnosed or refractory/relapse acute myeloid leukemia. Two cross-validation rounds were performed in seven nationwide reference laboratories in order to reach a consensus regarding quality metrics criteria and variant reporting. In the pre-standardization cross-validation round, an overall concordance of 60.98% was obtained with a great variability in selected genes and conditions across laboratories. After consensus of relevant genes and optimization of quality parameters the overall concordance rose to 85.57% in the second cross-validation round. We show that a diagnostic network with harmonized next-generation sequencing analysis and reporting in seven experienced laboratories is feasible in the context of a scientific group. This cooperative nationwide strategy provides advanced molecular diagnostic for acute myeloid leukemia patients of the PETHEMA group.
Alzheimer’s disease (AD) may manifest retinal changes preceding brain pathology. A transversal case-control study utilized spectral-domain OCT angiography (SD-OCTA) and Angio-Tool software 0.6a to ...assess retinal vascular structures and OCT for inner and outer retina thickness in the APPNL-F/NL-F AD model at 6, 9, 12, 15, 17, and 20 months old. Comparisons to age-matched wild type (WT) were performed. The analysis focused on the three vascular plexuses using AngiooTool and on retinal thickness, which was represented with the Early Treatment Diabetic Retinopathy Study (ETDRS) sectors. Compared to WT, the APPNL-F/NL-F group exhibited both vascular and structural changes as early as 6 months persisting and evolving at 15, 17, and 20 months. Significant vascular alterations, principally in the superficial vascular complex (SVC), were observed. There was a significant decrease in the vessel area and the total vessel length in SVC, intermediate, and deep capillary plexus. The inner retina in the APPNL-F/NL-F group predominantly decreased in thickness while the outer retina showed increased thickness in most analyzed time points compared to the control group. There are early vascular and structural retinal changes that precede the cognitive changes, which appear at later stages. Therefore, the natural history of the APPNL-F/NL-F model may be more similar to human AD than other transgenic models.
Toxic elements, such as mercury (Hg) and arsenic (As), are major pollutants in aquatic environments, posing ecological threats to living organisms due to their toxicity and bioaccumulation. This ...paper investigated whether zebrafish response to Hg and As displayed day/night differences. Fish were exposed to either 35 μg/L of mercury chloride for 6 h or 65 mg/L of sodium arsenate for 4 h, at two different times of the day: mid-light (day; ML) and mid-darkness (night; MD). Fish were video-recorded to investigate their behavioural response and at the end of each trial, gills and liver samples were collected for gene expression measurement. Gills, liver and brain samples were also obtained to determine Hg and As concentration. A control group (non-exposed) was video-recorded and sampled too. The effect of Hg and As on zebrafish swimming activity and the expression of antioxidant and metallothionein genes was time-of-day-dependent, with a stronger response being observed during the day than at night. However, the neurobehavioural effect of Hg was more affected by the time of exposure than the effect of As. In addition, Hg concentration in the gills was significantly higher in zebrafish exposed at ML than at MD. Altogether, these findings suggest that zebrafish response to Hg and As is time-of-day-dependent and remark the importance of considering toxicity rhythms when using this fish species as a model in toxicological research.
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•Zebrafish response to Hg and As exhibited day/night differences.•Hg affected activity levels in zebrafish exposed during the day but not at night.•Transcriptome responses to both Hg and As were mostly stronger during the day.•Hg concentration in zebrafish gills was higher after exposure during the day.
In Alzheimer’s disease (AD), transgenic mouse models have established links between abnormalities in the retina and those in the brain. APPNL-F/NL-F is a murine, humanized AD model that replicates ...several pathological features observed in patients with AD. Research has focused on obtaining quantitative parameters from optical coherence tomography (OCT) in AD. The aim of this study was to analyze, in a transversal case-control study using manual retinal segmentation via SD-OCT, the changes occurring in the retinal layers of the APPNL/F-NF/L AD model in comparison to C57BL/6J mice (WT) at 6, 9, 12, 15, 17, and 20 months of age. The analysis focused on retinal thickness in RNFL-GCL, IPL, INL, OPL, and ONL based on the Early Treatment Diabetic Retinopathy Study (ETDRS) sectors. Both APPNL-F/NL-F-model and WT animals exhibited thickness changes at the time points studied. While WT showed significant changes in INL, OPL, and ONL, the AD model showed changes in all retinal layers analyzed. The APPNL-F/NL-F displayed significant thickness variations in the analyzed layers except for the IPL compared to related WT. These thickness changes closely resembled those found in humans during preclinical stages, as well as during mild and moderate AD stages, making this AD model behave more similarly to the disease in humans.
An enhanced database (RetroDB) of the Spanish wind energy resources, derived from a high spatial resolution integration with the WRF model, is proposed and evaluated. RetroDB provides hourly capacity ...factor (CF) values for the Spanish regions, along the period of 2007–2020, with an unprecedented spatial resolution. RetroDB estimates were benchmarked based on the ERA5 global reanalysis. A comprehensive evaluation study of both RetroDB and ERA5 estimates was conducted using surface and tall mast measurements, along with actual CF values. The extent to which RetroDB and ERA5 reproduced the CF spatial variability, distribution, and ramp distribution were specifically addressed. The results showed no differences between the global and regional reanalysis performance regarding nationally aggregated wind energy estimates. Nevertheless, RetroDB clearly shows a superior performance reproducing the wind speeds’ and CFs’ spatial and temporal distributions. This was found to be related to the higher reliability of RetroDB reproducing the aloft winds in complex topographic areas. Overall, the results clearly indicate that, in areas such as the study region, where the wind resources are mostly associated with topographic enhancements, high spatial resolution regional reanalyses are preferable over relative coarse reanalyses (e.g., ERA5), particularly for wind energy integration studies. RetroDB database is made publicly available.
Reduced or null expression of E-cadherin protein is a frequent cause of diffuse gastric cancer (DGC). More than 50% of patients with DGC present somatic variants in
gene.
The objectives of this study ...were to study E-cadherin expression and identify variants in the
gene in gastric tumors of patients with DGC.
We studied 18 Mexican DGC patients who attended a hospital of the Mexican Social Security Institute; E-cadherin expression was determined by immunohistochemistry, and variants were identified by Sanger sequencing in promoter and coding regions. Predictive analysis was performed using PolyPhen-2 and HOPE software.
We found that 56% of DGC patients showed reduced expression of E-cadherin. All patients carried
variants; overall, 12 different
variants were identified. Predictive analysis revealed that the rs114265540 variant was probably damaging, with a value of 0.985, indicating a functional impact on the E-cadherin protein. Variants rs34939176 and rs33964119 were identified as risk factors for DGC (odds' ratios OR = 31.3, 95% CI 6.3-154.0, p < 0.001; OR = 6.1, 95% CI 2.0-19.0, p < 0.001, respectively) given their elevated frequency and by comparing it with those reported for MXL population in the 1000 Genomes Project database.
In this Mexican population, the percentage of diffuse gastric tumors with reduced expression of E-cadherin was similar to that reported in other populations. All gastric tumors of DGC patients studied had somatic
gene variants; however, the rs114265540, rs34939176, and rs33964119 variants were importantly related to DGC.
Posttranslational histone modifications and the dynamics of histone variant H2A.Z are key mechanisms underlying the floral transition. In yeast, SWR1-C and NuA4-C mediate the deposition of H2A.Z and ...the acetylation of histone H4, H2A and H2A.Z, respectively. Yaf9 is a subunit shared by both chromatin-remodeling complexes. The significance of the two Arabidopsis YAF9 homologues, YAF9A and YAF9B, is unknown.
To get an insight into the role of Arabidopsis YAF9 proteins in plant developmental responses, we followed physiological, genetic, genomic, epigenetic, proteomics and cell biology approaches.
Our data revealed that YAF9A and YAF9B are histone H3 readers with unequally redundant functions. Double mutant yaf9a yaf9b plants display pleiotropic developmental phenotypic alterations as well as misregulation of a wide variety of genes. We demonstrated that YAF9 proteins regulate flowering time by both FLC-dependent and independent mechanisms that work in parallel with SWR1-C. Interestingly, we show that YAF9A binds FLC chromatin and that YAF9 proteins regulate FLC expression by modulating the acetylation levels of H2A.Z and H4 but not H2A.Z deposition.
Our work highlights the key role exerted by YAF9 homologues in the posttranslational modification of canonical histones and variants that regulate gene expression in plants to control development.
Key points
Neurotransmitter release requires a tight coupling between synaptic vesicle exocytosis and endocytosis with dynamin being a key protein in that process.
We used imaging techniques to ...examine the time course of endocytosis at mouse motor nerve terminals expressing synaptopHluorin, a genetically encoded reporter of the synaptic vesicle cycle.
We separated two sequential phases of endocytosis taking place during the stimulation train: early and late endocytosis.
Freshly released synaptic vesicle proteins are preferentially retrieved during the early phase, which is very sensitive to dynasore, an inhibitor of dynamin GTPase activity.
Synaptic vesicle proteins pre‐existing at the plasma membrane before the stimulation are preferentially retrieved during the late phase, which is very sensitive to myristyl trimethyl ammonium bromide (MitMAB), an inhibitor of the dynamin–phospholipid interaction.
Synaptic endocytosis is essential at nerve terminals to maintain neurotransmitter release by exocytosis. Here, at the neuromuscular junction of synaptopHluorin (spH) transgenic mice, we have used imaging to study exo‐ and endocytosis occurring simultaneously during nerve stimulation. We observed two endocytosis components, which occur sequentially during stimulation. The early component of endocytosis apparently internalizes spH molecules freshly exocytosed. This component was sensitive to dynasore, a blocker of dynamin 1 GTPase activity. In contrast, this early component was resistant to myristyl trimethyl ammonium bromide (MiTMAB), a competitive agent that blocks dynamin binding to phospholipid membranes. The late component of endocytosis is likely to internalize spH molecules that pre‐exist at the plasma membrane before stimulation starts. This component was blocked by MiTMAB, perhaps by impairing the binding of dynamin or other key endocytic proteins to phospholipid membranes. Our study suggests the co‐existence of two sequential synaptic endocytosis steps taking place during stimulation that are susceptible to pharmacological dissection: an initial step, preferentially sensitive to dynasore, that internalizes vesicular components immediately after they are released, and a MiTMAB‐sensitive step that internalizes vesicular components pre‐existing at the plasma membrane surface. In addition, we report that post‐stimulus endocytosis also has several components with different sensitivities to dynasore and MiTMAB.
El término Industria 4.0 se ha convertido en prioridad y objeto de estudio para empresas y centros de investigación pero aún se encuentra dentro de sus primeras etapas de implementación. Además, las ...compañías enfrentan dificultades al desarrollar soluciones para Industria 4.0, sin estar seguras de cómo afrontar sus requerimientos básicos. El diseño de una arquitectura de referencia aborda explícitamente este problema, apoya a los profesionales en la implementación de soluciones siendo la base del desarrollo y proporciona un soporte ante los desafíos que la Industria 4.0 representa. Por lo tanto, la contribución de este documento se centra en diseñar una arquitectura de referencia para sistemas y aplicaciones en Industria 4.0 basada en computación en la nube y análisis de datos, mostrando su viabilidad a través de la implementación en un caso de uso: Agricultura 4.0. Mediante esta arquitectura, ingenieros e investigadores podrán enfrentar los desafíos actuales de la producción inteligente, así como investigar, desarrollar e implementar soluciones (aplicaciones y sistemas) guiadas, estandarizadas y a costos accesibles, que cumplan los requerimientos que gobiernan Industria 4.0.
Breast cancer expressing the estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor-2 (HER2) is known as triple-positive (TPBC). TPBC represents 9–11% of breast ...cancer cases worldwide and is a heterogeneous subtype. Notably, TPBC presents a therapeutic challenge due to the crosstalk between the hormonal (ER and PR) and HER2 pathways. Patients with TPBC are treated with trastuzumab (TTZ); however, several patients treated with TTZ tend to relapse. The present study aimed to investigate the effect of the PR on inhibitory effect of TTZ on cell viability. BT474 cells (a model of TPBC) and BT474 PR-silenced cells were treated with either TTZ, progesterone (Pg), the PR antagonist mifepristone (RU486) or estradiol (E2) alone or in combination for 144 h (6 days). Cell viability assays and western blotting were subsequently performed. The results showed that Pg and E2 interfered with the inhibitory effect of TTZ on cell viability and this effect was potentiated when both hormones were combined. Pg was revealed to act through the PR, mainly activating the PR isoform B (PR-B) and inducing the protein expression levels of CDK4 and cyclin D1; however, it did not reactivate the HER2/Akt pathway. By contrast, E2 was able to increase PR isoform A (PR-A) expression, which was inhibited by Pg. Notably, in most of the experiments, RU486 did not antagonize the effects of Pg. In conclusion, Pg and E2 may interfere with the inhibitory effect of TTZ on cell viability through PR-B activation and PR-A inactivation.
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