Hippocampal Pyk2 regulates specific social skills: Implications for schizophrenia.

Pyk2 has been shown previously to be involved in several psychological and cognitive alterations related to stress, Huntington's disease, and Alzheimer's disease. All these disorders are accompanied by different types of impairments in sociability, which has recently been linked to improper mitochondrial function. We hypothesize that Pyk2, which regulates mitochondria, could be associated with the regulation of mitochondrial dynamics and social skills. In the present manuscript, we report that a reduction of Pyk2 levels in mouse pyramidal neurons of the hippocampus decreased social dominance and aggressivity. Furthermore, social interactions induced robust Pyk2-dependent hippocampal changes in several oxidative phosphorylation complexes. We also observed that Pyk2 levels were increased in the CA1 pyramidal neurons of schizophrenic subjects, occurring alongside changes in different direct and indirect regulators of mitochondrial function including DISC1 and Grp75. Accordingly, overexpressing Pyk2 in hippocampal CA1 pyramidal cells mimicked some specific schizophrenia-like social behaviors in mice. In summary, our results indicate that Pyk2 might play a role in regulating specific social skills likely via mitochondrial dynamics and that there might be a link between Pyk2 levels in hippocampal neurons and social disturbances in schizophrenia.

Epidemiological and clinical management aspects of pneumonias diagnosed in the emergency department in elderly patients in Spain: Results of the EDEN-29 study.

OBJECTIVES: To estimate the incidence of pneumonia diagnosis in elderly patients in Spanish emergency departments (ED), need for hospitalization, adverse events and predictive capacity of biomarkers commonly used in the ED. METHODS: Patients ≥65 years with pneumonia seen in 52 Spanish EDs were included. We recorded in-hospitaland 30-day mortality as adverse events, as well as intensive care unit (ICU) admission among hospitalizedpatients. Association of 10 predefined variables with adverse events was calculated and expressed as odds ratio (OR) with 95% confidence interval (CI), as well as predictive capacity of 5 commonly used biomarkers in the ED (leukocytes, hemoglobin, C-reactive protein, glucose, creatinine) was investigated using area under the receiver operating characteristic curve (AUC-ROC). RESULTS: 591 patients with pneumonia attended in the ED were included (annual incidence of 18,4 per 1000 inhabitants). A total of 78.0% were hospitalized. Overall, 30-day mortality was 14.2% and in-hospital mortality was 12.9%. Functional dependency was associated with both events (OR=4.453, 95%CI=2.361-8.400; and OR=3.497, 95%CI=1.578-7.750, respectively) as well as severe comorbidity (2.344, 1.363-4.030, and 2.463, 1.252-4.846, respectively). Admission to the ICU during hospitalization occurred in 3.5%, with no associated factors. The predictive capacity of biomarkers was only moderate for creatinine for ICU admission (AUC-ROC=0.702, 95% CI=0.536-0.869) and for leukocytes for post-discharge adverse event (0.669, 0.540-0.798). CONCLUSIONS: Pneumonia is a frequent diagnosis in elderly patients consulting in the ED. Their functional dependence and comorbidity is the factor most associated with adverse events. The biomarkers analyzed do not have a good predictive capacity for adverse events.

Sex- and age-related patterns in the use of analgesics in older patients in the emergency department.

BACKGROUND: Treatment of acute pain in older patients is a common challenge faced in emergency departments (EDs). Despite many studies that have investigated chronic analgesic use in the elderly, data on patterns of acute use, especially in EDs, of analgesics according to patient characteristics is scarce. OBJECTIVE: To investigate sex- and age-related patterns of analgesic use in the Spanish EDs and determine differences in age-related patterns according to patient sex. DESIGN: A secondary analysis of the Emergency Department and Elderly Needs (EDEN) multipurpose cohort. SETTING: Fifty-two Spanish EDs (17% of Spanish EDs covering 25% of Spanish population). PARTICIPANTS: All patients' ≥65 years attending ED during 1 week (April 1-7, 2019). Patient characteristics recorded included age, sex, chronic treatment with non-steroidal anti-inflammatory drugs (NSAIDs) and opiates, comorbidity, dependence, dementia, depression, ability to walk and previous falls. Analgesics used in the ED were categorized in three groups: non-NSAID non-opioids (mainly paracetamol and metamizole, PM), NSAIDs, and opiates. OUTCOME MEASURES: Frequency of analgesic use was quantified, and the relationship between sex and age and analgesic use (in general and for each analgesic group) was assessed by unadjusted and adjusted logistic regression and restricted cubic spline models. Interaction between sex and age was explored. MAIN RESULTS: We included 24 573 patients, and 6678 (27.2%) received analgesics in the ED: 5551 (22.6%) PM, 1661 (6.8%) NSAIDs and 937 (3.8%) opiates (1312 received combinations). Analgesics were more frequently used in women (adjusted OR = 1.076, 95%CI = 1.014-1.142), as well as with NSAID (1.205, 1.083-1.341). Analgesic use increased with age, increasing PM and decreasing NSAIDs use. Opiate use remained quite constant across age and sex. Interaction of sex with age was present for the use of analgesics in general ( P  = 0.006), for PM ( P  < 0.001) and for opiates ( P  = 0.033), with higher use of all these analgesics in women. CONCLUSION: Use of analgesics in older individuals in EDs is mildly augmented in women and increases with age, with PM use increasing and NSAIDs decreasing with age. Conversely, opiate use is quite constant according to sex and age. Age-related patterns differ according to sex, with age-related curves of women showing higher probabilities than those of men to receive any analgesic, PM or opiates.

Delirium risk screening and assessment among older patients in general wards and the emergency department: a best practice implementation project.

OBJECTIVES: The aim of this project was to improve compliance with evidence-based criteria regarding risk of delirium and the assessment of delirium among older patients in the general hospitalization wards and the emergency department. INTRODUCTION: More than 50% of older hospitalized patients experience delirium. Some studies have highlighted the need to implement an orientation protocol in the emergency department and to continue this in the general wards, with the aim of decreasing the delirium rate among older patients admitted to hospital. METHODS: The project followed the JBI evidence implementation framework. We conducted a baseline audit, a half-way audit, and final audit of 50 patients at risk of delirium admitted to the emergency department and the general wards, respectively. The audits measured compliance with eight criteria informed by the available evidence. RESULTS: In the final audit, three of the eight criteria achieved more than 50% compliance in the general wards: pressure injury screening (96%); monitoring changes (74%); and performing interventions (76%). In the emergency department, worse results were reported because of the service conditions. The exception was the criterion on the training of nurses on the topic, with 98%. The integration of a tool to screen for delirium in older patients in the hospital's electronic clinical history records increased the percentage of compliance with audit criteria regarding the use of the scale and delirium detection (rising from 0% to 32% in the final audit in the general wards). CONCLUSION: Through the implementation of this project, validated and evidence-based evaluation will ensure that nurses are supported through appropriate measures to reduce patient confusion and aggression resulting from delirium.

[Estudio cuasi-experimental sobre intervenciones enfermeras en la valoración, manejo y prevención del síndrome confusional agudo.] / Quasi-experimental study on nursing interventions in the assessment, management and prevention of acute confusional syndrome.

OBJECTIVE: Acute confusional syndrome (ACS) is one of the complications with the highest morbidity and mortality in hospitalization units, but it is a reversible situation if detected early, representing a clear challenge for nursing. The objectives of this study were to assess the interventions carried out by nurses for the identification and non-pharmacological preventive measures applied in acute confusional syndrome and relate them to the years of professional experience and training received. METHODS: A quasi-experimental, prospective and analytical study was carried out through a self-administered structured questionnaire pre-post intervention (extracted from the JBI PACES program-Practical Application of Clinical Evidence System) on the identification and preventive measures applied in ACS. A total of 520 questionnaires (pre and post assessment) were distributed to nurses from the emergency department and the internal medicine unit of the Miguel Servet University Hospital in Zaragoza (Aragón, Spain) from January 2021 to April 2022. Statistical analysis carried out with the program Jamovi®2.3.13. RESULTS: 180 correctly completed questionnaires (94 pre and 86 post) were received. For 100%, the ACS supposed an extra workload and significant differences were found between the ability to manage ACS with the years of professional experience (p≤0.028). 97.2% of the nurses applied non-pharmacological interventions. CONCLUSIONS: Despite being perceived as an extra burden in daily work, nurses perform non-pharmacological prevention for the management of ACS. It is necessary to improve training to provide guidance strategies.

OBJETIVO: El síndrome confusional agudo (SCA) es una de las complicaciones con mayor morbimortalidad en las unidades de hospitalización, pero es una situación reversible si se detecta a tiempo, representando un claro desafío para la enfermería. Los objetivos de este estudio fueron valorar previa y posteriormente las intervenciones realizadas por las enfermeras para la identificación y la adopción de las medidas preventivas no farmacológicas aplicadas en el síndrome confusional agudo, así como relacionarlas con los años de experiencia profesional y la formación recibida. METODOS: Se realizó un estudio cuasi experimental, prospectivo y analítico a través de cuestionario estructurado autoadministrado pre-post intervención (extraído de la JBI PACES program-Practical Application of Clinical Evidence System) sobre la identificación y las medidas preventivas aplicadas en el SCA. Se distribuyeron un total de quinientos veinte cuestionarios (valoración pre y post) a enfermeras del servicio de Urgencias y la unidad de medicina interna del Hospital Universitario Miguel Servet de Zaragoza (Aragón, España) de enero de 2021 a abril de 2022. El análisis estadístico se realizó con el programa Jamovi® 2.3.13. RESULTADOS: Se recibieron ciento ochenta cuestionarios cumplimentados correctamente (noventa y cuatro pre y ochenta y seis post). Para el 100%, el SCA supuso una carga de trabajo extra y se hallaron diferencias estadísticamente significativas entre la capacidad de manejo de SCA con los años de experiencia profesional (p≤0,028). El 97,2% de las enfermeras aplicaron intervenciones no farmacológicas. CONCLUSIONES: A pesar de percibirse como una carga extra en el trabajo diario, las enfermeras realizan prevenciones no farmacológicas para el manejo del SCA. Es necesario mejorar la formación para proporcionar estrategias de orientación.

Development of Beta-Amyloid-Specific CAR-Tregs for the Treatment of Alzheimer's Disease.

BACKGROUND: Alzheimer's disease (AD) is a neurodegenerative disease that remains uncured. Its pathogenesis is characterized by the formation of ß-amyloid (Aß) plaques. The use of antigen-specific regulatory T cells (Tregs) through adoptive transfer has shown promise for the treatment of many inflammatory diseases, although the effectiveness of polyspecific Tregs is limited. Obtaining a sufficient number of antigen-specific Tregs from patients remains challenging. AIMS AND METHODS: To address this problem, we used an antibody-like single-chain variable fragment from a phage library and subsequently generated a chimeric antigen receptor (CAR) targeting ß-amyloid. RESULTS: The ß-amyloid-specific CARs obtained were stimulated by both recombinant and membrane-bound Aß isolated from the murine brain. The generated CAR-Tregs showed a normal Treg phenotype, were antigen-specific activatable, and had suppressive capacity. CONCLUSION: This study highlights the potential of CAR technology to generate antigen-specific Tregs and presents novel approaches for developing functional CARs.

Loss of TAZ after YAP deletion severely impairs foregut development and worsens cholestatic hepatocellular injury.

BACKGROUND: We previously showed that loss of yes-associated protein 1 (YAP) in early liver development (YAPKO) leads to an Alagille syndrome-like phenotype, with failure of intrahepatic bile duct development, severe cholestasis, and chronic hepatocyte adaptations to reduce liver injury. TAZ, a paralog of YAP, was significantly upregulated in YAPKO hepatocytes and interacted with TEA domain family member (TEAD) transcription factors, suggesting possible compensatory activity. METHODS: We deleted both Yap1 and Wwtr1 (which encodes TAZ) during early liver development using the Foxa3 promoter to drive Cre expression, similar to YAPKO mice, resulting in YAP/TAZ double knockout (DKO) and YAPKO with TAZ heterozygosity (YAPKO TAZHET). We evaluated these mice using immunohistochemistry, serum biochemistry, bile acid profiling, and RNA sequencing. RESULTS: DKO mice were embryonic lethal, but their livers were similar to YAPKO, suggesting an extrahepatic cause of death. Male YAPKO TAZHET mice were also embryonic lethal, with insufficient samples to determine the cause. However, YAPKO TAZHET females survived and were phenotypically similar to YAPKO mice, with increased bile acid hydrophilicity and similar global gene expression adaptations but worsened the hepatocellular injury. TAZ heterozygosity in YAPKO impacted the expression of canonical YAP targets Ctgf and Cyr61, and we found changes in pathways regulating cell division and inflammatory signaling correlating with an increase in hepatocyte cell death, cell cycling, and macrophage recruitment. CONCLUSIONS: YAP loss (with or without TAZ loss) aborts biliary development. YAP and TAZ play a codependent critical role in foregut endoderm development outside the liver, but they are not essential for hepatocyte development. TAZ heterozygosity in YAPKO livers increased cell cycling and inflammatory signaling in the setting of chronic injury, highlighting genes that are especially sensitive to TAZ regulation.

The GRP78-PERK axis contributes to memory and synaptic impairments in Huntington's disease R6/1 mice.

Increasing evidence indicates that a key factor in neurodegenerative diseases is the activation of the unfolded protein response (UPR) caused by an accumulation of misfolded proteins in the endoplasmic reticulum (ER stress). Particularly, in Huntington's disease (HD) mutant huntingtin (mHtt) toxicity involves disruption of the ER-associated degradation pathway and loss of the ER protein homeostasis leading to neuronal dysfunction and degeneration. Besides the role of the UPR in regulating cell survival and death, studies that demonstrate the contribution of sustained UPR activation, particularly of PERK signaling, in memory disturbances and synaptic plasticity deficiencies are emerging. Given the contribution of hippocampal dysfunction to emotional and cognitive deficits seen in HD, we have analyzed the involvement of ER stress in HD memory alterations. We have demonstrated that at early disease stages, ER stress activation manifested as an increase in GRP78 and CHOP is observed in the hippocampus of R6/1 mice. Genetic reduction of GRP78 expression resulted in preventing hippocampal-dependent memory alterations but no motor deficits. Accordingly, hippocampal neuropathology namely, dendritic spine loss and accumulation of mHtt aggregates was ameliorated by GRP78 reduction. To elucidate the signaling pathways, we found that the inactivation of PERK by GSK2606414 restored spatial and recognition memories in R6/1 mice and rescued dendritic spine density in CA1 pyramidal neurons and protein levels of some specific immediate early genes. Our study unveils the critical role of the GRP78/PERK axis in memory impairment in HD mice and suggests the modulation of PERK activation as a novel therapeutic target for HD intervention.

Generation of Chimeric Antigen Receptors against Tetraspanin 7.

Adoptive transfer of antigen-specific regulatory T cells (Tregs) has shown promising results in the treatment of autoimmune diseases; however, the use of polyspecific Tregs has limited effects. However, obtaining a sufficient number of antigen-specific Tregs from patients with autoimmune disorders remains challenging. Chimeric antigen receptors (CARs) provide an alternative source of T cells for novel immunotherapies that redirect T cells independently of the MHC. In this study, we aimed to generate antibody-like single-chain variable fragments (scFv) and subsequent CARs against tetraspanin 7 (TSPAN7), a membrane protein highly expressed on the surface of pancreatic beta cells, using phage display technology. We established two methods for generating scFvs against TSPAN7 and other target structures. Moreover, we established novel assays to analyze and quantify their binding abilities. The resulting CARs were functional and activated specifically by the target structure, but could not recognize TSPAN7 on the surface of beta cells. Despite this, this study demonstrates that CAR technology is a powerful tool for generating antigen-specific T cells and provides new approaches for generating functional CARs.

PD-1/CTLA-4 Blockade Leads to Expansion of CD8+PD-1int TILs and Results in Tumor Remission in Experimental Liver Cancer.

Background: Checkpoint inhibitors act on exhausted CD8+ T cells and restore their effector function in chronic infections and cancer. The underlying mechanisms of action appear to differ between different types of cancer and are not yet fully understood. Methods: Here, we established a new orthotopic HCC model to study the effects of checkpoint blockade on exhausted CD8+ tumor-infiltrating lymphocytes (TILs). The tumors expressed endogenous levels of HA, which allowed the study of tumor-specific T cells. Results: The induced tumors developed an immune-resistant TME in which few T cells were found. The few recovered CD8+ TILs were mostly terminally exhausted and expressed high levels of PD-1. PD-1/CTLA-4 blockade resulted in a strong increase in the number of CD8+ TILs expressing intermediate amounts of PD-1, also called progenitor-exhausted CD8+ TILs, while terminally exhausted CD8+ TILs were almost absent in the tumors of treated mice. Although transferred naïve tumor-specific T cells did not expand in the tumors of untreated mice, they expanded strongly after treatment and generated progenitor-exhausted but not terminally exhausted CD8+ TILs. Unexpectedly, progenitor-exhausted CD8+ TILs mediated the antitumor response after treatment with minimal changes in their transcriptional profile. Conclusion: In our model, few doses of checkpoint inhibitors during the priming of transferred CD8+ tumor-specific T cells were sufficient to induce tumor remission. Therefore, PD-1/CTLA-4 blockade has an ameliorative effect on the expansion of recently primed CD8+ T cells while preventing their development into terminally exhausted CD8+ TILs in the TME. This finding could have important implications for future T-cell therapies.

Environmental factors affecting rapid shear in fibers from the passage of a bullet.

In forensic casework, examination of garment damage can provide insight into the mechanism of the specific cause of fiber failure. Different methods of damage impart differing physical characteristics on individual fibers. These alterations are determined by a multitude of factors, among them increased temperature of affected fibers. The process of rapid shear occurs in thermoplastic materials following high-speed impact. It results in distinct features caused by excessive heat generated through the interaction, which is unable to dissipate at a rate that would leave the fibers unchanged. Rapid shear characteristics can be differentiated from other fracture patterns through non-destructive microscopical methods and with a minimal sample size. Fabric samples were shot under heated, chilled, and water-saturated conditions, using ammunition of varying velocities. Analyses performed on the defects were conducted using stereomicroscopy, polarized light microscopy and scanning electron microscopy. Globular-shaped fiber ends, characteristics attributed specifically to rapid shear, were observed in all nylon samples. Through this study, it was determined that the environmental conditions employed did not affect fiber end changes associated with rapid shear.

A cationic motif upstream Engrailed2 homeodomain controls cell internalization through selective interaction with heparan sulfates.

Engrailed2 (En2) is a transcription factor that transfers from cell to cell through unconventional pathways. The poorly understood internalization mechanism of this cationic protein is proposed to require an initial interaction with cell-surface glycosaminoglycans (GAGs). To decipher the role of GAGs in En2 internalization, we have quantified the entry of its homeodomain region in model cells that differ in their content in cell-surface GAGs. The binding specificity to GAGs and the influence of this interaction on the structure and dynamics of En2 was also investigated at the amino acid level. Our results show that a high-affinity GAG-binding sequence (RKPKKKNPNKEDKRPR), upstream of the homeodomain, controls En2 internalization through selective interactions with highly-sulfated heparan sulfate GAGs. Our data underline the functional importance of the intrinsically disordered basic region upstream of En2 internalization domain, and demonstrate the critical role of GAGs as an entry gate, finely tuning homeoprotein capacity to internalize into cells.

Potter Deformation Sequence Caused by 17q12 Deletion: A Lethal Constellation.

17q12 deletion syndrome causes developmental abnormalities of the kidneys, pancreas, genital tract, and neurodevelopment, and it has a wide range of phenotypes ranging from fetal demise to normal adulthood with minimal renal impairment. Here we describe a rare case of 17q12 deletion diagnosed prenatally, complicated by anhydramnios and Potter sequence. The baby was born but necessitated life-saving interventions due to pulmonary and renal insufficiency and ultimately succumbed to multi-organ failure. We present full autopsy results describing findings linked to 17q12 deletion, including severe bilateral multicystic renal dysplasia, pancreatic hypoplasia, and cysts adjacent to the Fallopian tubes. We also describe pulmonary hypoplasia and Potter facies as consequences of anhydramnios. We correlate these findings to our current understanding of molecular signals altered by 17q12 deletion, notably affecting HNF1B and LHX1 genes, which are known to mediate renal and genitourinary tract development.

Splenectomy induces biochemical remission and regeneration in experimental murine autoimmune hepatitis.

Autoimmune hepatitis (AIH) is a chronic immune-mediated inflammatory liver disease. It is known that AIH originates not from the spleen but from the liver itself. Nonetheless, most details of the etiology and pathophysiology are unknown. We induced experimental murine AIH (emAIH) in NOD/Ltj mice by single administration of a replication-deficient adenovirus and performed splenectomy during late-stage disease. Biochemical disease remission occurred, which was characterized by improvement in transaminase levels. The causes of this remission included a shift in the transcriptomic signature of serum proteins toward regeneration. At the cellular level, there was a marked decrease in activated CD8+ T cells and an increase in intrahepatic regulatory T cells (Tregs). Here, intrahepatic Treg numbers correlated with biochemical remission. Notably, an imbalance in the T-cell/B-cell ratio was observed, with a disproportionate increase in total B cells. In summary, intrahepatic increases in Tregs, biochemical remission, and regeneration could be induced by splenectomy in the late stage of emAIH.

YAP1 activation and Hippo pathway signaling in the pathogenesis and treatment of intrahepatic cholangiocarcinoma.

Intrahepatic cholangiocarcinoma (iCCA), the second most common primary liver cancer, is a highly lethal epithelial cell malignancy exhibiting features of cholangiocyte differentiation. iCCAs can potentially develop from multiple cell types of origin within liver, including immature or mature cholangiocytes, hepatic stem cells/progenitor cells, and from transdifferentiation of hepatocytes. Understanding the molecular mechanisms and genetic drivers that diversely drive specific cell lineage pathways leading to iCCA has important biological and clinical implications. In this context, activation of the YAP1-TEAD dependent transcription, driven by Hippo-dependent or -independent diverse mechanisms that lead to the stabilization of YAP1 is crucially important to biliary fate commitment in hepatobiliary cancer. In preclinical models, YAP1 activation in hepatocytes or cholangiocytes is sufficient to drive their malignant transformation into iCCA. Moreover, nuclear YAP1/TAZ is highly prevalent in human iCCA irrespective of the varied etiology, and significantly correlates with poor prognosis in iCCA patients. Based on the ubiquitous expression and diverse physiologic roles for YAP1/TAZ in the liver, recent studies have further revealed distinct functions of active YAP1/TAZ in regulating tumor metabolism, as well as the tumor immune microenvironment. In the current review, we discuss our current understanding of the various roles of the Hippo-YAP1 signaling in iCCA pathogenesis, with a specific focus on the roles played by the Hippo-YAP1 pathway in modulating biliary commitment and oncogenicity, iCCA metabolism, and immune microenvironment. We also discuss the therapeutic potential of targeting the YAP1/TAZ-TEAD transcriptional machinery in iCCA, its current limitations, and what future studies are needed to facilitate clinical translation.

Undercarboxylated Osteocalcin: A Promising Target for Early Diagnosis of Cardiovascular and Glycemic Disorders in Patients with Metabolic Syndrome: A Pilot Study.

Lifestyle changes are causing an exponential increase in the prevalence of obesity and metabolic syndrome (MetS) worldwide. The most frequent complications of these are the development of diabetes (T2D) and cardiovascular disease (CVD). Accurate tools are needed to classify the cardiovascular risk (CVR) in the MetS population. In recent years, numerous biomarkers of bone metabolism have been associated with CVR. The aim of this study was to determine the levels of undercarboxylated osteocalcin (ucOC) in a cohort of patients with MetS and to analyse its association with MetS parameters and CVR as well as with T2D prevalence. A longitudinal study was conducted in which a MetS population was followed for one year. Weight change, adherence to the Mediterranean diet (MedDiet), ucOC levels, MetS parameters and CVR were analysed and CVR was calculated using different scores. Our results showed a decrease of CVR associated with a better adherence to the MetDiet resulting in higher HDL-C and ucOC levels though the improvement of MetS risk factors. This bone protein appeared as a potential biomarker to classify CVR in the MetS population, especially for MetS patients without prevalent T2D. Furthermore, ucOC serum levels could be good predictors of T2D prevalence.

Hepatitis E virus genotype 3 microbiological surveillance by the Spanish Reference Laboratory: geographic distribution and phylogenetic analysis of subtypes from 2009 to 2019.

BackgroundHepatitis E virus genotype 3 (HEV-3) is widely distributed throughout Europe, with incidence of infections increasing in many countries. Belgium, Bulgaria, France, Germany, Italy, the Netherlands and the United Kingdom have reported the distribution of HEV-3 subtypes in cohorts of patients with hepatic disease.AimTo describe the distribution of the HEV-3 subtypes in Spain at national and autonomous community (AC) levels between 2009 and 2019. The study was also extended to Andorra.MethodsOf 5,197 samples received by the National Reference Laboratory during the study, 409 were HEV-RNA-positive. Among these, 294 (71.9%) were further typed based on an ORF2 sequence fragment, or, for a subset of 74, based on the full-coding genome sequence.ResultsHEV-3 was detected in 291 samples. The dominant subtype in Spain was HEV-3f (88.3%; 257/291), which occurred in all ACs, with no change in detection level over time. Within this subtype, three subclusters were characterised: HEV-3f-B, HEV-3f-A1 and HEV-3f-A2. The second most common HEV subtype was the recently described HEV-3m (7%; 21/291), with two subclusters identified: HEV-3m-A, which has been known since 2010, and HEV-3m-B, since 2014. The third most encountered subtype was HEV-3c (4.1%; 12/291), with a frequency not increasing over time, unlike observations in some European countries.ConclusionThe importance of the surveillance of HEV-3 subtype and subcluster circulation is yet to be assessed. This surveillance together with the comprehensive epidemiological characterisation of clinical cases, could support the identification of sources of transmission and the establishment of control measures nationally and internationally.

NOTCH-YAP1/TEAD-DNMT1 Axis Drives Hepatocyte Reprogramming Into Intrahepatic Cholangiocarcinoma.

BACKGROUND & AIMS: Intrahepatic cholangiocarcinoma (ICC) is a devastating liver cancer with extremely high intra- and inter-tumoral molecular heterogeneity, partly due to its diverse cellular origins. We investigated clinical relevance and the molecular mechanisms underlying hepatocyte (HC)-driven ICC development. METHODS: Expression of ICC driver genes in human diseased livers at risk for ICC development were examined. The sleeping beauty and hydrodynamic tail vein injection based Akt-NICD/YAP1 ICC model was used to investigate pathogenetic roles of SRY-box transcription factor 9 (SOX9) and yes-associated protein 1 (YAP1) in HC-driven ICC. We identified DNA methyltransferase 1 (DNMT1) as a YAP1 target, which was validated by loss- and gain-of-function studies, and its mechanism addressed by chromatin immunoprecipitation sequencing. RESULTS: Co-expression of AKT and Notch intracellular domain (NICD)/YAP1 in HC yielded ICC that represents 13% to 29% of clinical ICC. NICD independently regulates SOX9 and YAP1 and deletion of either, significantly delays ICC development. Yap1 or TEAD inhibition, but not Sox9 deletion, impairs HC-to-biliary epithelial cell (BEC) reprogramming. DNMT1 was discovered as a novel downstream effector of YAP1-TEAD complex that directs HC-to-BEC/ICC fate switch through the repression of HC-specific genes regulated by master regulators for HC differentiation, including hepatocyte nuclear factor 4 alpha, hepatocyte nuclear factor 1 alpha, and CCAAT/enhancer-binding protein alpha/beta. DNMT1 loss prevented NOTCH/YAP1-dependent HC-driven cholangiocarcinogenesis, and DNMT1 re-expression restored ICC development following TEAD repression. Co-expression of DNMT1 with AKT was sufficient to induce tumor development including ICC. DNMT1 was detected in a subset of HCs and dysplastic BECs in cholestatic human livers prone to ICC development. CONCLUSION: We identified a novel NOTCH-YAP1/TEAD-DNMT1 axis essential for HC-to-BEC/ICC conversion, which may be relevant in cholestasis-to-ICC pathogenesis in the clinic.

Bi-allelic hydroxymethylbilane synthase inactivation defines a homogenous clinico-molecular subtype of hepatocellular carcinoma.

BACKGROUND & AIMS: Acute intermittent porphyria (AIP), caused by heterozygous germline mutations of the heme synthesis pathway enzyme HMBS (hydroxymethylbilane synthase), confers a high risk of hepatocellular carcinoma (HCC) development. Yet, the role of HMBS in liver tumorigenesis remains unclear. METHODS: Herein, we explore HMBS alterations in a large series of 758 HCC cases, including 4 patients with AIP. We quantify the impact of HMBS mutations on heme biosynthesis pathway intermediates and we investigate the molecular and clinical features of HMBS-mutated tumors. RESULTS: We identify recurrent bi-allelic HMBS inactivation, both in patients with AIP acquiring a second somatic HMBS mutation and in sporadic HCC with 2 somatic hits. HMBS alterations are enriched in truncating mutations, in particular in splice regions, leading to abnormal transcript structures. Bi-allelic HMBS inactivation results in a massive accumulation of its toxic substrate porphobilinogen and synergizes with CTNNB1-activating mutations, leading to the development of well-differentiated tumors with a transcriptomic signature of Wnt/ß-catenin pathway activation and a DNA methylation signature related to ageing. HMBS-inactivated HCC mostly affects females, in the absence of fibrosis and classical HCC risk factors. CONCLUSIONS: These data identify HMBS as a tumor suppressor gene whose bi-allelic inactivation defines a homogenous clinical and molecular HCC subtype. LAY SUMMARY: Heme (the precursor to hemoglobin, which plays a key role in oxygen transport around the body) synthesis occurs in the liver and involves several enzymes including hydroxymethylbilane synthase (HMBS). HMBS mutations cause acute intermittent porphyria, a disease caused by the accumulation of toxic porphyrin precursors. Herein, we show that HMBS inactivation is also involved in the development of liver cancers with distinct clinical and molecular characteristics.

Importance of the use of pre- and intra-operative imaging as a tool for planning foreign body removal in the floor of the mouth: a case report / Importancia del uso de la imagenología pre e intraoperatoria como herramienta para la planificación de remoción de cuerpo extraño en piso de boca: presentación de un caso

Introduction: Body piercings consist of small holes made with a needle in different parts of the skin or body to introduce a jewel or decorative element. In the oral cavity, most piercings are placed in the tongue. However, some complications may occur, and surgical techniques must be used for their removal. These complications present a certain degree of difficulty due to their position and may challenge the ability of the clinician to access the specific anatomical location. The different imaging techniques, from simple radiography to intraoperative techniques such as image intensifiers, have become an extremely useful tool for locating an object in the three dimensions of space, allowing safe location and extraction. Objective: The aim of this study is to report the case of a complication of a body piercing in the oral cavity and how the use of imaging was decisive for surgical planning and for the quick and effective resolution of the case. Material and Methods: A 14-year-old female patient came looking for treatment. Her mother reported the onset of the condition after the insertion of a needle-like metallic object while performing an artistic perforation in the lingual region. Since the girl was unable to extract the object, she sought medical advice at the Carlos Arvelo Military Hospital in Caracas, Venezuela. Subsequently, an imaging study was performed by means of a Computed Tomography to locate the metallic object. It was observed that the foreign body had migrated to the floor of the mouth/sublingual region, requiring the area to be surgically approached. It was also decided to use an intraoperative image intensifier. The removal of the object was performed satisfactorily. Conclusion: The extraction of foreign bodies placed in the lingual and sublingual region represents a challenge for the clinician due to the number of important anatomical structures that pass through that area. This makes clinicians plan their surgical removal using pre- and intraoperative imaging, to find a less traumatic location, reduce surgical time as well as the risk of damaging adjacent anatomical structures.

Introducción: Los body piercings consisten en producir perforaciones con una aguja en diferentes localizaciones de la piel con el fin de introducir una joya o elemento decorativo a través del agujero producido previamente. En la cavidad oral, la lengua es el sitio de mayor elección; sin embargo, en ciertas ocasiones suelen presentarse ciertas complicaciones, por lo cual se debe recurrir a técnicas quirúrgicas para su remoción presentando cierto grado de dificultad para su localización y la capacidad del clínico para acceder al espacio anatómico. Las diferentes técnicas imagenológicas desde una radiografía simple hasta técnicas intraoperatorias como los intensificadores de imágenes se han convertido en una herramienta sumamente útil para la ubicación de un objeto en las tres dimensiones del espacio, permitiendo una localización y extracción segura para el clínico y el paciente. Objetivo: El propósito de este trabajo es reportar un caso de una complicación de esta práctica en la región bucal y cómo el uso de la imagenología fue determinante para la planificación quirúrgica y la resolución del caso de manera rápida y efectiva. Material y Métodos: Se presenta a consulta una paciente femenina de 14 años de edad, quien madre refiere inicio de enfermedad actual posterior a introducirse objeto metálico tipo aguja de compás con la finalidad de realizar perforación artística en región lingual, al no poder extraer dicho objeto, acude a evaluación en el Hospital Militar Carlos arvelo de Caracas; posterior se realiza estudio imagenológico tipo Tomografía Computarizada para la ubicación del objeto metálico donde se observa que el cuerpo extraño habría migrado hacia el piso de boca/región sublingual, teniendo la necesidad de abordar quirúrgicamente la zona y elegir el uso de intensificador de imagen transoperatorio para retiro del mismo el cual se dio de manera satisfactoria. Conclusión: Los cuerpos extraños desplazados a la región lingual y sublingual representan un desafío para el clínico al momento de extraerlos, esto se debe a la cantidad de estructuras anatómicas importantes que pasan por dicha zona, lo que hace que el clínico opte en su planificación quirúrgica por el uso de imagenología pre y transoperatoria obteniendo así una localización menos traumática, menor tiempo operatorio y menor riesgo de lesión de estructuras anatómicas adyacentes.