SATB2 organizes the 3D genome architecture of cognition in cortical neurons.

The DNA-binding protein SATB2 is genetically linked to human intelligence. We studied its influence on the three-dimensional (3D) epigenome by mapping chromatin interactions and accessibility in control versus SATB2-deficient cortical neurons. We find that SATB2 affects the chromatin looping between enhancers and promoters of neuronal-activity-regulated genes, thus influencing their expression. It also alters A/B compartments, topologically associating domains, and frequently interacting regions. Genes linked to SATB2-dependent 3D genome changes are implicated in highly specialized neuronal functions and contribute to cognitive ability and risk for neuropsychiatric and neurodevelopmental disorders. Non-coding DNA regions with a SATB2-dependent structure are enriched for common variants associated with educational attainment, intelligence, and schizophrenia. Our data establish SATB2 as a cell-type-specific 3D genome modulator, which operates both independently and in cooperation with CCCTC-binding factor (CTCF) to set up the chromatin landscape of pyramidal neurons for cognitive processes.

Machine learning combining multi-omics data and network algorithms identifies adrenocortical carcinoma prognostic biomarkers.

Background: Rare endocrine cancers such as Adrenocortical Carcinoma (ACC) present a serious diagnostic and prognostication challenge. The knowledge about ACC pathogenesis is incomplete, and patients have limited therapeutic options. Identification of molecular drivers and effective biomarkers is required for timely diagnosis of the disease and stratify patients to offer the most beneficial treatments. In this study we demonstrate how machine learning methods integrating multi-omics data, in combination with system biology tools, can contribute to the identification of new prognostic biomarkers for ACC. Methods: ACC gene expression and DNA methylation datasets were downloaded from the Xena Browser (GDC TCGA Adrenocortical Carcinoma cohort). A highly correlated multi-omics signature discriminating groups of samples was identified with the data integration analysis for biomarker discovery using latent components (DIABLO) method. Additional regulators of the identified signature were discovered using Clarivate CBDD (Computational Biology for Drug Discovery) network propagation and hidden nodes algorithms on a curated network of molecular interactions (MetaBase™). The discriminative power of the multi-omics signature and their regulators was delineated by training a random forest classifier using 55 samples, by employing a 10-fold cross validation with five iterations. The prognostic value of the identified biomarkers was further assessed on an external ACC dataset obtained from GEO (GSE49280) using the Kaplan-Meier estimator method. An optimal prognostic signature was finally derived using the stepwise Akaike Information Criterion (AIC) that allowed categorization of samples into high and low-risk groups. Results: A multi-omics signature including genes, micro RNA's and methylation sites was generated. Systems biology tools identified additional genes regulating the features included in the multi-omics signature. RNA-seq, miRNA-seq and DNA methylation sets of features revealed a high power to classify patients from stages I-II and stages III-IV, outperforming previously identified prognostic biomarkers. Using an independent dataset, associations of the genes included in the signature with Overall Survival (OS) data demonstrated that patients with differential expression levels of 8 genes and 4 micro RNA's showed a statistically significant decrease in OS. We also found an independent prognostic signature for ACC with potential use in clinical practice, combining 9-gene/micro RNA features, that successfully predicted high-risk ACC cancer patients. Conclusion: Machine learning and integrative analysis of multi-omics data, in combination with Clarivate CBDD systems biology tools, identified a set of biomarkers with high prognostic value for ACC disease. Multi-omics data is a promising resource for the identification of drivers and new prognostic biomarkers in rare diseases that could be used in clinical practice.

MAP kinase ERK5 modulates cancer cell sensitivity to extrinsic apoptosis induced by death-receptor agonists.

Death receptor ligand TRAIL is a promising cancer therapy due to its ability to selectively trigger extrinsic apoptosis in cancer cells. However, TRAIL-based therapies in humans have shown limitations, mainly due inherent or acquired resistance of tumor cells. To address this issue, current efforts are focussed on dissecting the intracellular signaling pathways involved in resistance to TRAIL, to identify strategies that sensitize cancer cells to TRAIL-induced cytotoxicity. In this work, we describe the oncogenic MEK5-ERK5 pathway as a critical regulator of cancer cell resistance to the apoptosis induced by death receptor ligands. Using 2D and 3D cell cultures and transcriptomic analyses, we show that ERK5 controls the proteostasis of TP53INP2, a protein necessary for full activation of caspase-8 in response to TNFα, FasL or TRAIL. Mechanistically, ERK5 phosphorylates and induces ubiquitylation and proteasomal degradation of TP53INP2, resulting in cancer cell resistance to TRAIL. Concordantly, ERK5 inhibition or genetic deletion, by stabilizing TP53INP2, sensitizes cancer cells to the apoptosis induced by recombinant TRAIL and TRAIL/FasL expressed by Natural Killer cells. The MEK5-ERK5 pathway regulates cancer cell proliferation and survival, and ERK5 inhibitors have shown anticancer activity in preclinical models of solid tumors. Using endometrial cancer patient-derived xenograft organoids, we propose ERK5 inhibition as an effective strategy to sensitize cancer cells to TRAIL-based therapies.

R-loop landscapes in the developing human brain are linked to neural differentiation and cell-type specific transcription.

Here, we construct genome-scale maps for R-loops, three-stranded nucleic acid structures comprised of a DNA/RNA hybrid and a displaced single strand of DNA, in the proliferative and differentiated zones of the human prenatal brain. We show that R-loops are abundant in the progenitor-rich germinal matrix, with preferential formation at promoters slated for upregulated expression at later stages of differentiation, including numerous neurodevelopmental risk genes. RNase H1-mediated contraction of the genomic R-loop space in neural progenitors shifted differentiation toward the neuronal lineage and was associated with transcriptomic alterations and defective functional and structural neuronal connectivity in vivo and in vitro. Therefore, R-loops are important for fine-tuning differentiation-sensitive gene expression programs of neural progenitor cells.

Homologs of Ancestral CNNM Proteins Affect Magnesium Homeostasis and Circadian Rhythmicity in a Model Eukaryotic Cell.

Biological rhythms are ubiquitous across organisms and coordinate key cellular processes. Oscillations of Mg2+ levels in cells are now well-established, and due to the critical roles of Mg2+ in cell metabolism, they are potentially fundamental for the circadian control of cellular activity. The identity of the transport proteins responsible for sustaining Mg2+ levels in eukaryotic cells remains hotly debated, and several are restricted to specific groups of higher eukaryotes. Here, using the eukaryotic minimal model cells of Ostreococcus tauri, we report two homologs of common descents of the Cyclin M (CNNM)/CorC protein family. Overexpression of these proteins leads to a reduction in the overall magnesium content of cells and a lengthening of the period of circadian gene expression rhythms. However, we observed a paradoxical increase in the magnesium content of the organelle fraction. The chemical inhibition of Mg2+ transport has a synergistic effect on circadian period lengthening upon the overexpression of one CNNM homolog, but not the other. Finally, both homologs rescue the deleterious effect of low extracellular magnesium on cell proliferation rates. Overall, we identified two CNNM proteins that directly affect Mg2+ homeostasis and cellular rhythms.

The ERK5/NF-κB signaling pathway targets endometrial cancer proliferation and survival.

Endometrial cancer (EC) is the most common type of gynecologic cancer in women of developed countries. Despite surgery combined with chemo-/radiotherapy regimens, overall survival of patients with high-risk EC tumors is poor, indicating a need for novel therapies. The MEK5-ERK5 pathway is activated in response to growth factors and to different stressors, including oxidative stress and cytokines. Previous evidence supports a role for the MEK5-ERK5 pathway in the pathology of several cancers. We investigated the role of ERK5 in EC. In silico analysis of the PanCancer Atlas dataset showed alterations in components of the MEK5-ERK5 pathway in 48% of EC patients. Here, we show that ERK5 inhibition or silencing decreased EGF-induced EC cell proliferation, and that genetic deletion of MEK5 resulted in EC impaired proliferation and reduced tumor growth capacity in nude mice. Pharmacologic inhibition or ERK5 silencing impaired NF-kB pathway in EC cells and xenografts. Furthermore, we found a positive correlation between ERK5 and p65/RELA protein levels in human EC tumor samples. Mechanistically, genetic or pharmacologic impairment of ERK5 resulted in downregulation of NEMO/IKKγ expression, leading to impaired p65/RELA activity and to apoptosis in EC cells and xenografts, which was rescued by NEMO/IKKγ overexpression. Notably, ERK5 inhibition, MEK5 deletion or NF-kB inhibition sensitized EC cells to standard EC chemotherapy (paclitaxel/carboplatin) toxicity, whereas ERK5 inhibition synergized with paclitaxel to reduce tumor xenograft growth in mice. Together, our results suggest that the ERK5-NEMO-NF-κB pathway mediates EC cell proliferation and survival. We propose the ERK5/NF-κB axis as new target for EC treatment.

Out of Africa: Juvenile Dispersal of Black-Shouldered Kites in the Emerging European Population.

Knowledge of animal dispersal patterns is of great importance for the conservation and maintenance of natural populations. We here analyze juvenile dispersal of the poorly studied Black-shouldered Kite (Elanus caeruleus) monitored in southwestern Spain in an ongoing long-term study initiated in 2003. The European population of Black-shouldered kites is thought to be a recent one funded by colonizing African birds, as no kites have been found in the European fossil record, and the breeding population has progressively expanded to the North in the late 20th and 21st centuries. We obtained information on movements behavior during dispersal from 47 juveniles Kites after marking 384 nestlings with wing tags and three nestlings with radio transmitter. We have tested two competing hypotheses (i.e., the Resources Competition Hypothesis and the Wandering Hypothesis (WH)) that may explain the leptokurtic distribution of the natal dispersal distance in Elanus. After independence, juvenile females dispersed farther from the natal areas than males, as is common in birds. On average, males and females dispersed from their natal areas over 9 (i.e., 26.15 km) and 15 (i.e., 43.79 km) breeding territories, respectively. A male and two females dispersed further than 100 km from their natal nest. Our results indicated some evidence supporting the competition-for-resources hypotheses since nestlings hatched from high quality territories stayed closer from natal areas than nestlings hatched from low quality territories and also nestlings hatched first within the brood also tend to recruit closer to their natal area than later hatched nestlings which tend to disperse further away from their natal area. The information provided by these crucial demographic parameters will be used for the elaboration of future conservation plans for the management of this colonizing species in Europe.

Neuron-specific chromosomal megadomain organization is adaptive to recent retrotransposon expansions.

Regulatory mechanisms associated with repeat-rich sequences and chromosomal conformations in mature neurons remain unexplored. Here, we map cell-type specific chromatin domain organization in adult mouse cerebral cortex and report strong enrichment of Endogenous Retrovirus 2 (ERV2) repeat sequences in the neuron-specific heterochromatic B2NeuN+ megabase-scaling subcompartment. Single molecule long-read sequencing and comparative Hi-C chromosomal contact mapping in wild-derived SPRET/EiJ (Mus spretus) and laboratory inbred C57BL/6J (Mus musculus) reveal neuronal reconfigurations tracking recent ERV2 expansions in the murine germline, with significantly higher B2NeuN+ contact frequencies at sites with ongoing insertions in Mus musculus. Neuronal ablation of the retrotransposon silencer Kmt1e/Setdb1 triggers B2NeuN+ disintegration and rewiring with open chromatin domains enriched for cellular stress response genes, along with severe neuroinflammation and proviral assembly with infiltration of dendrites . We conclude that neuronal megabase-scale chromosomal architectures include an evolutionarily adaptive heterochromatic organization which, upon perturbation, results in transcriptional dysregulation and unleashes ERV2 proviruses with strong neuronal tropism.

ERK5 Inhibition Induces Autophagy-Mediated Cancer Cell Death by Activating ER Stress.

Autophagy is a highly conserved intracellular process that preserves cellular homeostasis by mediating the lysosomal degradation of virtually any component of the cytoplasm. Autophagy is a key instrument of cellular response to several stresses, including endoplasmic reticulum (ER) stress. Cancer cells have developed high dependency on autophagy to overcome the hostile tumor microenvironment. Thus, pharmacological activation or inhibition of autophagy is emerging as a novel antitumor strategy. ERK5 is a novel member of the MAP kinase family that is activated in response to growth factors and different forms of stress. Recent work has pointed ERK5 as a major player controlling cancer cell proliferation and survival. Therefore small-molecule inhibitors of ERK5 have shown promising therapeutic potential in different cancer models. Here, we report for the first time ERK5 as a negative regulator of autophagy. Thus, ERK5 inhibition or silencing induced autophagy in a panel of human cancer cell lines with different mutation patterns. As reported previously, ERK5 inhibitors (ERK5i) induced apoptotic cancer cell death. Importantly, we found that autophagy mediates the cytotoxic effect of ERK5i, since ATG5-/- autophagy-deficient cells viability was not affected by these compounds. Mechanistically, ERK5i stimulated autophagic flux independently of the canonical regulators AMPK or mTORC1. Moreover, ERK5 inhibition resulted in ER stress and activation of the Unfolded Protein Response (UPR) pathways. Specifically, ERK5i induced expression of the ER luminal chaperone BiP (a hallmark of ER stress), the UPR markers CHOP and ATF4, and the spliced form of XBP1. Pharmacological inhibition of UPR with chemical chaperone TUDC, or ATF4 silencing, resulted in impaired ERK5i-mediated UPR, autophagy and cytotoxicity. Overall, our results suggest that ERK5 inhibition induces autophagy-mediated cancer cell death by activating ER stress. Since ERK5 inhibition sensitizes cancer cells and tumors to chemotherapy, future work will determine the relevance of UPR and autophagy in the combined use of chemotherapy and ERK5i to tackle Cancer.

Influence of the Computer-Aided Static Navigation Technique and Mixed Reality Technology on the Accuracy of the Orthodontic Micro-Screws Placement. An In Vitro Study.

To analyze the effect of a computer-aided static navigation technique and mixed reality technology on the accuracy of orthodontic micro-screw placement. Material and methods: Two hundred and seven orthodontic micro-screws were placed using either a computer-aided static navigation technique (NAV), a mixed reality device (MR), or a conventional freehand technique (FHT). Accuracy across different dental sectors was also analyzed. CBCT and intraoral scans were taken both prior to and following orthodontic micro-screw placement. The deviation angle and horizontal deviation were then analyzed; these measurements were taken at the coronal entry point and apical endpoint between the planned and performed orthodontic micro-screws. In addition, any complications resulting from micro-screw placement, such as spot perforations, were also analyzed across all dental sectors. Results: The statistical analysis showed significant differences between study groups with regard to the coronal entry-point (p < 0.001). The NAV study group showed statistically significant differences from the FHT (p < 0.001) and MR study groups (p < 0.001) at the apical end-point (p < 0.001), and the FHT group found significant differences from the angular deviations of the NAV (p < 0.001) and MR study groups deviations (p = 0.0011). Different dental sectors also differed significantly. (p < 0.001) Additionally, twelve root perforations were observed in the FHT group, while there were no root perforations in the NAV group. Conclusions: Computer-aided static navigation technique enable more accurate orthodontic micro-screw placement and fewer intraoperative complications when compared with the mixed reality technology and conventional freehand techniques.

Deep-coverage spatiotemporal proteome of the picoeukaryote Ostreococcus tauri reveals differential effects of environmental and endogenous 24-hour rhythms.

The cellular landscape changes dramatically over the course of a 24 h day. The proteome responds directly to daily environmental cycles and is additionally regulated by the circadian clock. To quantify the relative contribution of diurnal versus circadian regulation, we mapped proteome dynamics under light:dark cycles compared with constant light. Using Ostreococcus tauri, a prototypical eukaryotic cell, we achieved 85% coverage, which allowed an unprecedented insight into the identity of proteins that facilitate rhythmic cellular functions. The overlap between diurnally- and circadian-regulated proteins was modest and these proteins exhibited different phases of oscillation between the two conditions. Transcript oscillations were generally poorly predictive of protein oscillations, in which a far lower relative amplitude was observed. We observed coordination between the rhythmic regulation of organelle-encoded proteins with the nuclear-encoded proteins that are targeted to organelles. Rhythmic transmembrane proteins showed a different phase distribution compared with rhythmic soluble proteins, indicating the existence of a circadian regulatory process specific to the biogenesis and/or degradation of membrane proteins. Our observations argue that the cellular spatiotemporal proteome is shaped by a complex interaction between intrinsic and extrinsic regulatory factors through rhythmic regulation at the transcriptional as well as post-transcriptional, translational, and post-translational levels.

Distribución del género Tityus Koch, 1836 (Scorpiones: Buthidae) en la ciudad de La Plata (Argentina) / Distribution of the genus Tityus Koch, 1836 (Scorpiones: Buthidae) in the city of La Plata (Argentina)

Resumen En la ciudad de La Plata se encuentra el género Tityus con registros desde 1943. Se estudió la distribución territorial del género Tityus en esta ciudad y su expansión en los últimos 15 años a partir de las consultas recibidas en el Laboratorio de Aracnología del CEPAVE, provenientes de particulares e instituciones públicas y privadas. Se recibieron 189 consultas desde el año 2005 al 2020. Se utilizó el programa QGIS para geolocalizar cada escorpión con capas de trazado urbano y desagües pluviales. Para el análisis espacial de distribución se consideró un home range de una hectárea por individuo para el cálculo de área y en los casos de áreas de influencias solapadas durante el mismo año se consideró el área como la unión de las mismas. El género Tityus en la ciudad de La Plata está representado por las especies T. carrilloi y T. confluens ambas de interés sanitario y de hábitos sinantrópicos, que ocupan dos zonas diferentes bien definidas. El análisis de la dispersión independiente en las dos zonas indicaría que podrían usar los desagües para dispersarse, y al ser inconexos estos no habría flujo de escorpiones entre ambas zonas. La colonización de estas especies en la zona en estudio se vio afianzada para T. confluens a partir del año 2005 y para T. carrilloi a partir del 2011. Los mayores registros son en los meses cálidos, desde enero a abril, siendo este último el de mayor valor. Septiembre es el único mes sin denuncia. La tasa de expansión calculada fue de 4.42 ha/año.

Abstract In the city of La Plata occurs the genus Tityus with records dating back to 1943. The territorial distribution of the genus Tityus in this city and its expansion was studied based on inquiries received at the CEPAVE Arachnology Laboratory from individuals and public and private institutions. 189 inquiries were received from 2005 to 2020. The QGIS program was used to geolocate each scorpion with urban layout layers and storm drains. For the spatial analysis of distribution, a home range of one hectare per individual was considered to calculate the area and in the cases of areas of overlapping influences during the same year, the area was considered as their union. The genus Tityus in the city of La Plata is represented by the species T. carrilloi and T. confluens, both of health interest and of synanthropic habits, which occupy two different well-defined areas. The analysis of the independent dispersion in the two zones would indicate that they could use the drains to disperse, and since they are unconnected there would be no flow of scorpions between both zones. The colonization of these species in the study area was strengthened for T. confluens as of 2005 and for T. carrilloi as of 2011. The highest records are in the warm months, from January to April, the latter being of higher value. September is the only month without complaints. The expansion rate calculated was 4.42 ha/year.

Influence of the Computer-Aided Static Navigation Technique on the Accuracy of the Orthodontic Micro-Screws Placement: An In Vitro Study.

To analyze the influence of the computer-aided static navigation technique on the accuracy of placement of orthodontic micro-screws. One hundred and thirty-eight orthodontic micro-screws were randomly assigned to the following study groups: Group A. orthodontic micro-screw placement using a computer-aided static navigation technique (n = 69); B. orthodontic micro-screw placement using the conventional freehand technique (n = 69). In addition, the accuracy in the canine-premolar, premolar and molar sectors was analyzed in each study group. Cone-beam computed tomography and intraoral scans were taken both prior and subsequent to orthodontic micro-screw placement. The images were then uploaded using a 3D implant planning software, where the deviation and horizontal angles were analyzed using a multivariate linear model. These measurements were taken at the coronal entry point and apical endpoint between the planned orthodontic micro-screws. In addition, any complications resulting from micro-screw placement, such as spot perforations, were also analyzed in all dental sectors. The statistical analysis showed significant differences between the two study groups with regard to the coronal entry-point, apical end-point (p < 0.001) and angular deviations (p < 0.001) between the computer-aided static navigation technique and freehand technique study groups. Moreover, statistically significant differences were showed between the different dental sectors (p < 0.001). Additionally, twelve root perforations were observed at the conventional free hand technique study group while there were no root perforations in the computer-aided static navigation technique study group. The results showed that the computer-aided static navigation technique enables a more accurate orthodontic micro-screw placement with less intraoperative complications when compared with the conventional freehand technique.

Sex differences in behavioral traits related with high sensitivity to the reinforcing effects of cocaine.

Cocaine is the most prevalent illegal stimulant drug in Europe among the adult population. Its abuse is characterized by a faster substance abuse disorder (SUD) development than other drugs, with high vulnerability to relapse. However, there does not exist an effective treatment for cocaine dependence. Sex differences have been reported in psychological disorders including SUD. For this reason, it is essential to identify risk factors that predict susceptibility or resilience to cocaine addiction for the development of effective prevention strategies considering sex differences. In the present study, the main objective was to determine more sensitive phenotypes to the conditioned reinforcing effects of cocaine in both sexes. Anxiety-like behavior and the locomotor response to novelty were evaluated in the elevated plus maze, and despair in the tail suspension test, as well as vulnerability traits linked with a high sensitivity to the reinforcing effects of a subthreshold dose of cocaine (1 mg/kg) in the conditioned place preference (CPP) paradigm in male and female mice. Our results indicated that only female mice with high anxiety, low locomotor response to novelty or low despair levels acquired CPP induced by cocaine, while male mice with low anxiety, high locomotor response to novelty or high despair levels presented a higher susceptibility to the rewarding effects of cocaine than others. These sex differences in the results reveal an opposite pattern in males and females on the relationship between anxiety- and depressive-like behaviors and cocaine vulnerability, demonstrating the need to include female mice in preclinical studies.

[Predictors of complex urethral surgery in anterior urethral stricture disease.] / Predictores de cirugía uretral compleja en estenosis de la uretra anterior.

OBJECTIVE: To determine the clinical-surgical factors associated with Complex Urethral Surgery (CUC) in anterior urethral stenosis. MATERIAL AND METHOD: This is a cross-sectional study with retrospective data recording, including all male patients who under went anterior urethroplasty between 2011 and 2018. CUC included two or more grafts urethroplasty, excisional augmented anastomotic urethroplasty, combined flaps and grafts urethroplasty and stages surgery. The data were collected from the electronic medical record, recording the demographic data, background of previous treatments as well as the characteristics of the stenosis (etiology, anatomical location, length, number of strictures, among others). A univariate and multivariate analysis were conducted using the chi-squared test and logistic regression to identify the variables related to CUC. RESULTS: The data of 665 patients met the inclusion criteria were analyzed. The mean age was 56.1 years, 27.5% were smokers, 32.5% had received some previous treatment, and dilatations were the most common procedure. The most prevalent etiology was iatrogenic, followed by idiopathic in a 61.1 and 20.3% respectively. Bulbar urethral stricture were the most common location (56.2%) while the mean length of the stenosis was 4.8 cm. After univariate and multivariate analysis, previous dilations (HR 2.6), multifocality (2.51), lengthof stenosis (>4 cm) (HR 1.49) and the hypospadias etiology (HR 11.9) were independent predictors for CUC (p<0.05)CONCLUSIONS: Hypospadias was the only etiology factor that predicts the need for CUC. Regarding radiological findings, extensive and multifocal stenosis, were predictors of complex surgery. History of previous dilations were also predictors of CUC.

OBJETIVO: Determinar los factores predictores asociados a Cirugía Uretral Compleja (CUC) en los casos de estenosis de la uretra anterior.MATERIAL Y MÉTODO: Estudio transversal con registro de datos retrospectivo, incluye a todos los pacientes masculinos a quienes se les practicó una plastía de la uretra anterior entre 2011 y 2018. Como CUC se consideró a la Uretroplastia con dos o más injertos, la anastomosis término terminal ampliada, la uretroplastia combinada y la cirugía por estadíos. Los datos se recabaron de la historia clínica electrónica consignándose aquellos demográficos, antecedente de tratamientos previos, así como las características de la estenosis (etiología, ubicación anatómica, longitud, número de estenosis entre otras). Se realizó un análisis univariado y multivariado para identificar variables predictoras de CUC. RESULTADOS: Se analizaron los datos de 665 pacientes con criterios de inclusión válidos. La media de edad fue de 56,1 años, 27,5% eran fumadores crónicos, 32,5% habían recibido algún tratamiento previo, siendo las dilataciones el procedimiento más común (17,6%), la etiología más prevalente fue la iatrógena seguida por la idiopática en un 61,1 y un 20,3% respectivamente, la ubicación más común fue la uretra bulbar en 56,2% mientras que la media de longitud de la estenosis fue de 4,8 cm. En el análisis univariado y multivariable el antecedente de dilataciones (HR 2,6), la multifocalidad (HR 2,51), la longitud de la estenosis (>4 cm) (HR 1,49) y la etiología hipospadias (HR 11,9) resultaron ser factores predictores independientes para CUC (p<0,05). CONCLUSIONES: La hipospadia fue dentro de las etiologías analizadas la única que predice la necesidad de CUC. En lo que respecta a antecedentes y hallazgos radiológicos, las dilataciones y las estenosis extensas y multifocales respectivamente fueron predictores de CUC.

Environmental Enrichment Induces Epigenomic and Genome Organization Changes Relevant for Cognition.

In early development, the environment triggers mnemonic epigenomic programs resulting in memory and learning experiences to confer cognitive phenotypes into adulthood. To uncover how environmental stimulation impacts the epigenome and genome organization, we used the paradigm of environmental enrichment (EE) in young mice constantly receiving novel stimulation. We profiled epigenome and chromatin architecture in whole cortex and sorted neurons by deep-sequencing techniques. Specifically, we studied chromatin accessibility, gene and protein regulation, and 3D genome conformation, combined with predicted enhancer and chromatin interactions. We identified increased chromatin accessibility, transcription factor binding including CTCF-mediated insulation, differential occupancy of H3K36me3 and H3K79me2, and changes in transcriptional programs required for neuronal development. EE stimuli led to local genome re-organization by inducing increased contacts between chromosomes 7 and 17 (inter-chromosomal). Our findings support the notion that EE-induced learning and memory processes are directly associated with the epigenome and genome organization.

The anti-cancer drug ABTL0812 induces ER stress-mediated cytotoxic autophagy by increasing dihydroceramide levels in cancer cells.

ABTL0812 is a first-in-class small molecule with anti-cancer activity, which is currently in clinical evaluation in a phase 2 trial in patients with advanced endometrial and squamous non-small cell lung carcinoma (NCT03366480). Previously, we showed that ABTL0812 induces TRIB3 pseudokinase expression, resulting in the inhibition of the AKT-MTORC1 axis and macroautophagy/autophagy-mediated cancer cell death. However, the precise molecular determinants involved in the cytotoxic autophagy caused by ABTL0812 remained unclear. Using a wide range of biochemical and lipidomic analyses, we demonstrated that ABTL0812 increases cellular long-chain dihydroceramides by impairing DEGS1 (delta 4-desaturase, sphingolipid 1) activity, which resulted in sustained ER stress and activated unfolded protein response (UPR) via ATF4-DDIT3-TRIB3 that ultimately promotes cytotoxic autophagy in cancer cells. Accordingly, pharmacological manipulation to increase cellular dihydroceramides or incubation with exogenous dihydroceramides resulted in ER stress, UPR and autophagy-mediated cancer cell death. Importantly, we have optimized a method to quantify mRNAs in blood samples from patients enrolled in the ongoing clinical trial, who showed significant increased DDIT3 and TRIB3 mRNAs. This is the first time that UPR markers are reported to change in human blood in response to any drug treatment, supporting their use as pharmacodynamic biomarkers for compounds that activate ER stress in humans. Finally, we found that MTORC1 inhibition and dihydroceramide accumulation synergized to induce autophagy and cytotoxicity, phenocopying the effect of ABTL0812. Given the fact that ABTL0812 is under clinical development, our findings support the hypothesis that manipulation of dihydroceramide levels might represents a new therapeutic strategy to target cancer.Abbreviations: 4-PBA: 4-phenylbutyrate; AKT: AKT serine/threonine kinase; ATG: autophagy related; ATF4: activating transcription factor 4; Cer: ceramide; DDIT3: DNA damage inducible transcript 3; DEGS1: delta 4-desaturase, sphingolipid 1; dhCer: dihydroceramide; EIF2A: eukaryotic translation initiation factor 2 alpha; EIF2AK3: eukaryotic translation initiation factor 2 alpha kinase 3; ER: endoplasmic reticulum; HSPA5: heat shock protein family A (Hsp70) member 5; MAP1LC3B: microtubule associated protein 1 light chain 3 beta; MEF: mouse embryonic fibroblast; MTORC1: mechanistic target of rapamycin kinase complex 1; NSCLC: non-small cell lung cancer; THC: Δ9-tetrahydrocannabinol; TRIB3: tribbles pseudokinase 3; XBP1: X-box binding protein 1; UPR: unfolded protein response.

In vitro evaluation of the dissolving effect of carbonated beverages (Coca-Cola®) and enzyme-based solutions on enteroliths obtained from horses: pilot study / Avaliação in vitro do efeito dissolvente de bebidas carbonatadas (Coca-Cola®) e soluções à base de enzimas sobre enterólitos obtidos de cavalos: estudo piloto

Enteroliths are concretions of minerals that cause partial or total obstruction of the intestinal lumen, resulting in recurrent and chronic colic in horses. This pilot study aimed to evaluate the in vitro solvent effect of carbonated beverages (Coca-Cola® and Coca-Cola® Zero), and papain and cellulase enzymes (Robinson Pharma®, Santa Ana, CA, USA) on enteroliths obtained from horses. Six 51-grams-samples of six enteroliths were assigned to six treatments of immersion solutions: T1, Coca-Cola®; T2: Coca-Cola® Zero; T3: distilled water + papain (90 mg) and cellulase (120 mg); T4: Coca-Cola® + papain and cellulase; T5: Coca-Cola® Zero + papain and cellulase; and, CT: distilled water (control). The volume for immersion in the assigned solution was 150 mL, at a pH of 7.1, using an incubation shaker (Heidolph®, Germany) at 37ºC and 25 rpm, for 72 h. The evaluation periods of the dissolution percentage (difference between the initial weight and final weight of the samples), were 0, 3, 12, 24, 36, 48, 60, and 72 h. After 72 h of immersion, solutions T4, T5, and T1 presented 47, 38.8, and 14.9% of dissolution, respectively. The other solutions did not have major differences with CT (control). Under the in vitro conditions of this pilot study, papain and cellulase enzymes potentiated the dissolving effect of the carbonated solutions on the enteroliths obtained from horses. Further studies are suggested since the existing literature is on the dissolution of phytobezoars and not of enteroliths.(AU)

Enterólitos são concreções de minerais que causam obstrução parcial ou total do lume intestinal, resultando em cólica crônica e recorrente nos cavalos. Este estudo piloto teve como objetivo avaliar in vitro o efeito dissolvente sobre os enterólitos das bebidas carbonatadas (Coca-Cola® e Coca-Cola® Zero) e a solução à base das enzimas papaína e celulase (Robinson Pharma®, Santa Ana, CA, USA). Seis (6) amostras de seis (6) enterólitos de 51gramas de peso foram distribuídas em seis tratamentos de imersão: T1: Coca-Cola®; T2: Coca-Cola® Zero; T3: água destilada + papaína (90 mg) e celulase (120 mg); T4: Coca-Cola® + papaína e celulase; T5: Coca-Cola® Zero + papaína e celulase; e, CT: água destilada (controle). O volume das soluções de imersão foi de 150 mL, com pH de 7.1, usando um shaker de incubação (Heidolph®, Germany) com 37ºC e 25 rpm, durante 72 horas. A avaliação dos períodos da porcentagem de dissolução (diferenças entre o peso inicial e o peso final das amostras) foram 0, 3, 12, 24, 36, 48, 60 e 72 h. Depois de 72 h de imersão, as soluções T4, T5 e T1 apresentaram 47, 38,8 e 14,9% de dissolução, respectivamente. As outras soluções não tiveram diferenças com relação ao CT (controle). Nas condições in vitro deste estudo piloto, as enzimas papaína e celulase potenciam o efeito dissolvente das bebidas carbonatadas sobre os enterólitos obtidos de cavalos. Mais estudos são sugeridos, uma vez que só existe literatura sobre a dissolução de fitobezoares e não de enterólitos.(AU)

Author Correction: The effector AWR5 from the plant pathogen Ralstonia solanacearum is an inhibitor of the TOR signalling pathway.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

STK11 (LKB1) missense somatic mutant isoforms promote tumor growth, motility and inflammation.

Elucidating the contribution of somatic mutations to cancer is essential for personalized medicine. STK11 (LKB1) appears to be inactivated in human cancer. However, somatic missense mutations also occur, and the role/s of these alterations to this disease remain unknown. Here, we investigated the contribution of four missense LKB1 somatic mutations in tumor biology. Three out of the four mutants lost their tumor suppressor capabilities and showed deficient kinase activity. The remaining mutant retained the enzymatic activity of wild type LKB1, but induced increased cell motility. Mechanistically, LKB1 mutants resulted in differential gene expression of genes encoding vesicle trafficking regulating molecules, adhesion molecules and cytokines. The differentially regulated genes correlated with protein networks identified through comparative secretome analysis. Notably, three mutant isoforms promoted tumor growth, and one induced inflammation-like features together with dysregulated levels of cytokines. These findings uncover oncogenic roles of LKB1 somatic mutations, and will aid in further understanding their contributions to cancer development and progression.