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Instituto de Salud Carlos III, Comunidad Autónoma de Madrid, Depto. de Biología Celular, Depto. de Fisiología, Fac. de Ciencias Biológicas, TRUE, pub

2018 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved. En esta publicación han participado: 1Department of Internal Medicine III, University Hospital, RWTH, Aachen, Germany; 2Department of Immunology, Ophtalmology & ORL,Complutense University School of Medicine, Madrid, Spain; 3 12 de Octubre Health Research Institute (imas12), Madrid, Spain; 4Department of Hepatobiliary Surgery, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing Jiangsu, China; 5Department of Anesthesiology and Pain Management, Shanghai East Hospital, Tongji University, Shanghai, China; 6Institute of Pathology, Braunschweig Hospital, Braunschweig, Germany, Background & Aims: Caspase 8 (CASP8) is the apical initiator caspase in death receptor-mediated apoptosis. Strong evidence for a link between death receptor signaling pathways and cholestasis has recently emerged. Herein, we investigated the role of CASP8-dependent and independent pathways during experimental cholestasis. Methods: Liver injury was characterized in a cohort of human sera (n = 28) and biopsies from patients with stage IV primary biliary cholangitis. In parallel, mice with either specific deletion of Casp8 in liver parenchymal cells (Casp8Dhepa) or hepatocytes (Casp8Dhep), and mice with constitutive Ripk3 (Ripk3 / ) deletion, were subjected to surgical ligation of the common bile duct (BDL) from 2 to 28 days. Floxed (Casp8fl/fl) and Ripk3+/+ mice were used as controls. Moreover, the pan-caspase inhibitor IDN-7314 was used, and cell death mechanisms were studied in primary isolated hepatocytes. Results: Overexpression of activated caspase 3, CASP8 and RIPK3 was characteristic of liver explants from patients with primary biliary cholangitis. Twenty-eight days after BDL, Casp8Dhepa mice showed decreased necrotic foci, serum aminotransferase levels and apoptosis along with diminished compensatory proliferation and ductular reaction. These results correlated with a decreased inflammatory profile and ameliorated liver fibrogenesis. A similar phenotype was observed in Ripk3-/- mice. IDN-7314 treatment decreased CASP8 levels but failed to prevent BDL-induced cholestasis, independently of CASP8 in hepatocytes. Conclusion: These findings show that intervention against CASP8 in liver parenchymal cells – specifically in cholangiocytes– might be a beneficial option for treating obstructive cholestasis, while broad pan-caspase inhibition might trigger undesirable side effects. Lay summary: Loss of caspase 8 – a protein involved in programmed cell death – in liver parenchymal cells protects against experimental cholestasis. Therefore, specific pharmacological interve, IZKF, Ministerio de Economía, Comercio y Empresa, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Objectives To investigate whether hypertension could modify the function of adrenergic, nitrergic, and sensory innervations involved in the electrical field stimulation-induced response in mesenteric arteries from female rats. Methods Vascular reactivity experiments were performed in endothelium-denuded mesenteric arteries from normotensive, Wistar–Kyoto and spontaneously hypertensive female rats; protein expression was measured by western blot; nitric oxide release was measured by fluorometry; calcitonin gene-related peptide and noradrenaline release were determined by enzyme immunoassay. Results The electrical field stimulation-induced contractions were significantly lower in segments from spontaneously hypertensive rats than those of Wistar–Kyoto rats. Hypertension did not modify either the response or release of noradrenaline. Preincubation with the nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester increased the electrical field stimulation-induced contractions only in segments from Wistar–Kyoto rats. The relaxation induced by the nitric oxide donor sodium nitroprusside was similar in segments from both strains. The electrical field stimulation-induced nitric oxide release was decreased in segments from spontaneously hypertensive rats. The calcitonin gene-related peptide receptor antagonist CGRP(8–37) did not alter the electrical field stimulation-induced contractions in segments from Wistar–Kyoto rats but increased them in segments from spontaneously hypertensive rats. The calcitonin gene-related peptide-induced relaxation was increased in segments from spontaneously hypertensive rats. The expression of the 15-kDa active form of RAMP1 was increased in segments from spontaneously hypertensive rats. Conclusion In contrast to male rats, electrical field stimulation-induced contractions are decreased in hypertensive female rats. Nitrergic innervation plays a role in the development and/or maintenance of hypertension, whereas sensory innervation is a, Direccion General de Ciencia y Tecnologia, Instituto de Salud Carlos III Spanish Government, Depto. de Biología Celular, Fac. de Odontología, TRUE, pub

Background & aims: E-cyclins control the transition of quiescent cells into the cell cycle. Two E-cyclins, CcnE1 and CcnE2, have been described, but their specific contributions to cell cycle reentry in vivo are poorly understood. Liver regeneration following partial hepatectomy is an excellent in vivo model for the study of cell cycle reentry of quiescent cells. We investigated the relevance of E-cyclins in directing resting hepatocytes into the cell cycle after partial hepatectomy using CcnE1 and CcnE2 knockout mice. Methods: Partial hepatectomy (70%) was performed in CcnE1 (E1(-/-)) and CcnE2 (E2(-/-)) knockout and wild-type mice. Liver regeneration was monitored by cell cycle markers for G(1)/S phase, S phase, and M phase as well as by determining the liver/body weight ratio after partial hepatectomy. Ploidy of hepatocytes was determined by fluorescence-activated cell sorting and fluorescent in situ hybridization. Results: CcnE1 deletion resulted in normal liver regeneration with a slight delay of the G(1)/S-phase transition and a defect in endoreplication of otherwise polyploid hepatocytes. Surprisingly, E2(-/-) mice displayed accelerated and sustained DNA synthesis after partial hepatectomy, excessive endoreplication in hepatocytes, and a liver mass that was 45% greater than that of wild-type mice after termination of the regeneration process. CcnE2 depletion induced overexpression of CcnE1 and prolonged cdk2 kinase activity after partial hepatectomy. Conclusions: CcnE2 has an unexpected role in repressing CcnE1; the phenotype of E2(-/-) mice appears to result from CcnE1 overexpression and cdk2 hyperactivation. CcnE1 and CcnE2 therefore have nonredundant functions for S-phase entry and endoreplication during liver regeneration., Deutsche Forschungsgemeinschaft (DFG), National Cancer Institute (NCI), Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Biliary excretion is the main route of disposal of bilirubin and impaired excretion results in jaundice, a well recognisable symptom of liver disease. Conjugation of bilirubin in the liver is essential for its clearance. The glucuronidation of bilirubin is catalysed by the microsomal UDP-glucuronosyltransferase UGT1A1. Patients with Crigler-Najjar syndrome type 1 and Gunn rats, mutant strain of the Wistar rats, bear an autosomal recessive disorder resulting in hyperbilirubinemia. The aim of this work is to add new data about activity of UGT1A1 during the perinatal period and adult life. The results showed that activity of UGT1A1 is detectable from day 22 of the gestation. After birth, activity of UGT1A1 gradually increases and reaches the levels of adult life. Furthermore, bilirubin azopigments have been separated and characterized by thin layer chromatography. We have found that concentration of samples by evaporation and ulterior storing at -20 degrees C seemed to be suitable for the maintenance of samples., Spanish Social Security Funds for Research, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Hepatocyte transplantation would offer an attractive alternative to liver transplantation in the treatment of inborn errors of liver metabolism. However, a major problem in most transplantation studies to date has been the limited growth of transplanted cells in the recipient organ. We performed a strategy for selective proliferation of transplanted cells by interfering with the proliferative capacity of resident hepatocytes, using the pyrrolizidine alkaloid retrorsine and then transplanting liver cells in conjunction with repeated administration of triiodothyronine, an inducer of hepatocyte proliferation in rats. In the present study, foetal and adult syngeneic hepatocyte transplantation into spleen was performed in retrorsine-treated hyperbilirubinemic Gunn rats. In parallel, repeated injections of triiodothyronine were given to recipients. Rats were sacrificed at 1, 7, 30 and 90 days after transplantation and blood and bile samples were taken to assess the functionality of transplanted cells. The proliferative activity of transplanted hepatocytes was evaluated using proliferating cell nuclear antigen labelling index. In summary, both adult and foetal hepatocyte transplantation were effective in correcting a metabolic abnormality in Gunn rats for as long as 3 months. The RS/T3 model, as a measure to increase graft function, could represent an important advance to future clinical application of hepatocyte transplantation., Spanish Social Security Funds for Research, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Instituciones participantes: 1Department of Internal Medicine III, University Hospital, RWTH Aachen; 2Department of Gastroenterology and Hepatology, University Hospital Duisburg-Essen, Essen, Germany; 3Nutrition, Metabolism & Genomics group, Wageningen University, Division of Human Nutrition, Wageningen, The Netherlands; 4Norwich Medical School, University of East Anglia, Norwich, United Kingdom; 5Proteomics Facility, University Hospital, RWTH Aachen; 6Institute of Pathology, University Hospital, RWTH Aachen, Germany; and 7Howard Hughes Medical Institute and University of Massachusetts Medical School, Worcester, Massachusetts, BACKGROUND & AIMS: c-Jun N-terminal kinase (JNK) 1 and JNK2 are expressed in hepatocytes and have overlapping and distinct functions. JNK proteins are activated via phosphorylation in response to acetaminophen- or carbon tetrachloride (CCl4)- induced liver damage; the level of activation correlates with the degree of injury. SP600125, a JNK inhibitor, has been reported to block acetaminophen-induced liver injury. We investigated the role of JNK in drug-induced liver injury (DILI) in liver tissue from patients and in mice with genetic deletion of JNK in hepatocytes. METHODS: We studied liver sections from patients with DILI (due to acetaminophen, phenprocoumon, nonsteroidal antiinflammatory drugs, or autoimmune hepatitis) or patients without acute liver failure (controls) collected from a DILI Biobank in Germany. Levels of total and activated (phosphorylated) JNK were measured by immunohistochemistry and Western blotting. Mice with hepatocyte-specific deletion of Jnk1 (Jnk1Dhepa) or combination of Jnk1 and Jnk2 (JnkDhepa), as well as Jnk1-floxed C57BL/6 (control) mice, were given injections of CCl4 (to induce fibrosis) or acetaminophen (to induce toxic liver injury). We performed gene expression microarray and phosphoproteomic analyses to determine mechanisms of JNK activity in hepatocytes. RESULTS: Liver samples from DILI patients contained more activated JNK, predominantly in nuclei of hepatocytes and in immune cells, than healthy tissue. Administration of acetaminophen to JnkDhepa mice produced a greater level of liver injury than that observed in Jnk1Dhepa or control mice, based on levels of serum markers and microscopic and histologic analysis of liver tissues. Administration of CCl4 also induced stronger hepatic injury in JnkDhepa mice, based on increased inflammation, cell proliferation, and fibrosis progression, compared with Jnk1Dhepa or control mice. Hepatocytes from JnkDhepa mice given acetaminophen had an increased oxidative stress response, leading to de, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Acetaminophen (APAP) hepatotoxicity induces endoplasmic reticulum (ER) stress which triggers the unfolded protein response (UPR) in hepatocytes. However, the mechanisms underlying ER stress remain poorly understood, thus reducing the options for exploring new pharmacological therapies for patients with hyperacute liver injury. Eight-to-twelve-week-old C57BL/6J Xbp1-floxed (Xbp1f/f) and hepatocyte-specific knockout Xbp1 mice (Xbp1∆hepa) were challenged with either high dose APAP [500 mg/kg] and sacrificed at early (1-2 h) and late (24 h) stages of hepatotoxicity. Histopathological examination of livers, immunofluorescence and immunohistochemistry, Western blot, real time (RT)-qPCR studies and transmission electron microscopy (TEM) were performed. Pharmacological inhibition of XBP1 using pre-treatment with STF-083010 [STF, 75 mg/kg] and autophagy induction with Rapamycin [RAPA, 8 mg/kg] or blockade with Chloroquine [CQ, 60 mg/kg] was also undertaken in vivo. Cytoplasmic expression of XBP1 coincided with severity of human and murine hyperacute liver injury. Transcriptional and translational activation of the UPR and sustained activation of JNK1/2 were major events in APAP hepatotoxicity, both in a human hepatocytic cell line and in a preclinical model. Xbp1∆hepa livers showed decreased UPR and JNK1/2 activation but enhanced autophagy in response to high dose APAP. Additionally, blockade of XBP1 splicing by STF, mitigated APAP-induced liver injury and without non-specific off-target effects (e.g., CYP2E1 activity). Furthermore, enhanced autophagy might be responsible for modulating CYP2E1 activity in Xbp1∆hepa animals. Genetic and pharmacological inhibition of Xbp1 specifically in hepatocytes ameliorated APAP-induced liver injury by enhancing autophagy and decreasing CYP2E1 expression. These findings provide the basis for the therapeutic restoration of ER stress and/or induction of autophagy in patients with hyperacute liver injury., Ministerio de Economía, Comercio y Empresa, German Research Foundation, Ministerio de Ciencia, Innovación y Universidades, Fundación Científica de la Asociación Española Contra el Cancer, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Department of Medicine. Division of Liver Diseases. Mount Sinai School of Medicine. New York, USA, Liver disease is a major cause of illness and death worldwide. A central component in the complex network leading to the development of alcoholic liver disease is the activation of Kupffer cells by endotoxin and other soluble mediators. Alcohol consumption induces a state of "leaky gut increasing plasma and liver endotoxin levels. When Kupffer cells become activated, they interact with a complex of proteins located on the extracellular membrane signaling to produce a wide array of soluble factors, including cytokines, chemokines, growth factors, cyclooxygenase and lipoxygenase metabolites, and reactive oxygen species such as superoxide anion, hydrogen peroxide, and nitric oxide, all of which provide physiologically diverse and pivotal paracrine effects on all other liver cell types and, ultimately, liver injury. Kupffer cells are also central to the liver homeostatic response to injury as upon cellular degenerative changes, they immediately respond to the insult and release mediators to orchestrate inflammatory and reparative responses. Thus, the homeostatic responses are initiated by Kupffer cell-derived mediators at the cellular level and underlie the liver s defense and reparative mechanisms against injury. In order to understand better the role of Kupffer cells in the onset of liver injury, animal models in which Kupffer cells are inactivated, and cell culture settings (e.g. co-cultures) are being used with promising results that advance our understanding of alcoholic liver disease., National Institute of Diabetes and Digestive and Kidney Diseases (US Public Health Service), Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

2016 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved. Instituciones participantes: 1Department of Internal Medicine III, University Hospital, RWTH Aachen, Aachen, Germany; 2Merck Serono Geneva Research Centre, Case postale 54, chemin des Mines 9, Geneva CH-1211 20, Switzerland; 3Dutch Molecular Pathology Center, Department of Pathobiology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3508 TB Utrecht, The Netherlands; 4University Medical Center Groningen, Department of Pediatrics, University of Groningen, NL-9713 Groningen, The Netherlands, Background & Aims: During liver inflammation, triggering fibrogenesis and carcinogenesis immune cells play a pivotal role. In the present study we investigated the role of CCL5 in human and in murine models of chronic liver inflammation leading to hepatocellular carcinoma (HCC) development. Methods: CCL5 expression and its receptors were studied in well-defined patients with chronic liver disease (CLD) and in two murine inflammation based HCC models. The role of CCL5 in inflammation, fibrosis, tumor initiation and progression was analyzed in different cell populations of NEMODhepa/CCL5-/- animals and after bone marrow transplantation (BMT). For therapeutic intervention Evasin-4 was injected for 24 h or 8 weeks. Results: In CLD patients, CCL5 and its receptor CCR5 are overexpressed – an observation confirmed in the Mdr2-/- and NEMODhepa model. CCL5 deletion in NEMODhepa mice diminished hepatocyte apoptosis, compensatory proliferation and fibrogenesis due to reduced immune cell infiltration. Especially, CD45+/Ly6G+granulocytes, CD45+/CD11b+/Gr1.1+/F4/80+ proinflammatory monocytes, CD4+ and CD8+ T cells were decreased. One year old NEMODhepa/CCL5-/- mice displayed smaller and less malignant tumors, characterized by reduced proliferative capacity and less pronounced angiogenesis. We identified hematopoietic cells as the main source of CCL5, while CCL5 deficiency did not sensitise NEMODhepa hepatocytes towards TNFa induced apoptosis. Finally, therapeutic intervention with Evasin-4 over a period of 8 weeks ameliorated liver disease progression. Conclusion: We identified an important role of CCL5 in human and functionally in mice with disease progression, especially HCC development. A novel approach to inhibit CCL5 in vivo thus appears encouraging for patients with CLD. Lay summary: Our present study identifies the essential role of the chemoattractive cytokine CCL5 for liver disease progression and especially hepatocellular carcinoma development in men and mice. Finally, IZKF (UKA, RWTH Aachen), Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Individuals exhibiting an intermediate alcohol drinking pattern in conjunction with signs of metabolic risk present clinical features of both alcohol-associated and metabolic-associated fatty liver diseases. However, such combination remains an unexplored area of great interest, given the increasing number of patients affected. In the present study, we aimed to develop a preclinical DUAL (alcohol-associated liver disease plus metabolic-associated fatty liver disease) model in mice. C57BL/6 mice received 10% vol/vol alcohol in sweetened drinking water in combination with a Western diet for 10, 23, and 52 weeks (DUAL model). Animals fed with DUAL diet elicited a significant increase in body mass index accompanied by a pronounced hypertrophy of adipocytes, hypercholesterolemia, and hyperglycemia. Significant liver damage was characterized by elevated plasma alanine aminotransferase and lactate dehydrogenase levels, extensive hepatomegaly, hepatocyte enlargement, ballooning, steatosis, hepatic cell death, and compensatory proliferation. Notably, DUAL animals developed lobular inflammation and advanced hepatic fibrosis. Sequentially, bridging cirrhotic changes were frequently observed after 12 months. Bulk RNA-sequencing analysis indicated that dysregulated molecular pathways in DUAL mice were similar to those of patients with steatohepatitis. Conclusion: Our DUAL model is characterized by obesity, glucose intolerance, liver damage, prominent steatohepatitis and fibrosis, as well as inflammation and fibrosis in white adipose tissue. Altogether, the DUAL model mimics all histological, metabolic, and transcriptomic gene signatures of human advanced steatohepatitis, and therefore serves as a preclinical tool for the development of therapeutic targets., Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

E-type cyclins E1 (CcnE1) and E2 (CcnE2) are regulatory subunits of cyclin-dependent kinase 2 (Cdk2) and thought to control the transition of quiescent cells into the cell cycle. Initial findings indicated that CcnE1 and CcnE2 have largely overlapping functions for cancer development in several tumor entities including hepatocellular carcinoma (HCC). In the present study, we dissected the differential contributions of CcnE1, CcnE2, and Cdk2 for initiation and progression of HCC in mice and patients. To this end, we tested the HCC susceptibility in mice with constitutive deficiency for CcnE1 or CcnE2 as well as in mice lacking Cdk2 in hepatocytes. Genetic inactivation of CcnE1 largely prevented development of liver cancer in mice in two established HCC models, while ablation of CcnE2 had no effect on hepatocarcinogenesis. Importantly, CcnE1-driven HCC initiation was dependent on Cdk2. However, isolated primary hepatoma cells typically acquired independence on CcnE1 and Cdk2 with increasing progression in vitro, which was associated with a gene signature involving secondary induction of CcnE2 and up-regulation of cell cycle and DNA repair pathways. Importantly, a similar expression profile was also found in HCC patients with elevated CcnE2 expression and poor survival. In general, overall survival in HCC patients was synergistically affected by expression of CcnE1 and CcnE2, but not through Cdk2. Our study suggests that HCC initiation specifically depends on CcnE1 and Cdk2, while HCC progression requires expression of any E-cyclin, but no Cdk2., German Research Foundation, Ramón y Cajal Fellowship, Ministerio de Economía, Comercio y Empresa. Retos, Genomics Facility and by the Confocal Microscopy Facility of the Interdisciplinary Center for Clinical Research (IZKF), Faculty of Medicine at RWTH Aachen University, Depto. de Inmunología, Oftalmología y ORL, Fac. de Medicina, TRUE, pub

Background. Portal vein embolization is performed weeks before extended hepatic resections to increase the future liver remnant and prevent posthepatectomy liver failure. Portal vein embolization performed closer to the operation also could be protective, but worsening of portal hyper-perfusion is a major concern. We determined the hepatic hemodynamic effects of a portal vein embolization performed 24 hours prior to hepatic operation. Methods. An extended (90%) hepatectomy was performed in swine undergoing (portal vein embolization) or not undergoing (control) a portal vein embolization 24 hours earlier (n = 10/group). Blood tests, hepatic and systemic hemodynamics, hepatic function (plasma disappearance rate of indocyanine green), liver histology, and volumetry (computed tomographic scanning) were assessed before and after the hepatectomy. Hepatocyte proliferating cell nuclear antigen expression and hepatic gene expression also were evaluated. Results. Swine in the control and portal vein embolization groups maintained stable systemic hemodynamics and developed similar increases of portal blood flow (302 ± 72% vs 486 ± 92%, P = .13). Portal pressure drastically increased in Controls (from 9.4 ± 1.3 mm Hg to 20.9 ± 1.4 mm Hg, P < .001), while being markedly attenuated in the portal vein embolization group (from 11.4 ± 1.5 mm Hg to 16.1 ± 1.3 mm Hg, P = .061). The procedure also improved the preservation of the hepatic artery blood flow, liver function, and periportal edema. These effects occurred in the absence of hepatocyte proliferation or hepatic growth and were associated with the induction of the vasoprotective gene Klf2. Conclusion. Portal vein embolization preconditioning represents a potential hepato-protective strategy for extended hepatic resections. Further preclinical studies should assess its medium-term effects, including survival. Our study also supports the relevance of hepatic hemodynamics as the main pathogenetic factor of post-hepatectomy liver fa, Mineco, ISCIII-Fondos FEDER, Sociedad Española de Trasplante Hepático, Depto. de Farmacología y Toxicología, Fac. de Veterinaria, TRUE, pub

Background Eosinophilic esophagitis (EoE) is a chronic non-IgE-mediated allergic disease of the esophagus. An unbiased proteomics approach was performed to investigate pathophysiological changes in esophageal epithelium. Additionally, an RNAseq-based transcriptomic analysis in paired samples was also carried out. Methods Total proteins were purified from esophageal endoscopic biopsies in a cohort of adult EoE patients (n = 25) and healthy esophagus controls (n = 10). Differentially accumulated (DA) proteins in EoE patients compared to control tissues were characterized to identify altered biological processes and signaling pathways. Results were also compared with a quantitative proteome dataset of the human esophageal mucosa. Next, results were contrasted with those obtained after RNAseq analysis in paired samples. Finally, we matched up protein expression with two EoE-specific mRNA panels (EDP and Eso-EoE panel). Results A total of 1667 proteins were identified, of which 363 were DA in EoE. RNA sequencing in paired samples identified 1993 differentially expressed (DE) genes. Total RNA and protein levels positively correlated, especially in DE mRNA-proteins pairs. Pathway analysis of these proteins in EoE showed alterations in immune and inflammatory responses for the upregulated proteins, and in epithelial differentiation, cornification and keratinization in those downregulated. Interestingly, a set of DA proteins, including eosinophil-related and secreted proteins, were not detected at the mRNA level. Protein expression positively correlated with EDP and Eso-EoE, and corresponded with the most abundant proteins of the human esophageal proteome. Conclusions We unraveled for the first time key proteomic features involved in EoE pathogenesis. An integrative analysis of transcriptomic and proteomic datasets provides a deeper insight than transcriptomic alone into understanding complex disease mechanisms., European Regional Development Fund, Ministerio de Ciencia e Innovación (MCIN), Ministerio de Trabajo y Economía Social, Comunidad de Madrid, Asociacion Española de Gastroenterología, Instituto de Salud Carlos III, Depto. de Biología Celular, Fac. de Ciencias Biológicas, TRUE, pub

(1) Background: Transition is a planned movement of paediatric patients to adult healthcare systems, and its implementation is not yet established in all inflammatory bowel disease (IBD) units. The aim of the study was to evaluate the impact of transition on IBD outcomes. (2) Methods: Multicentre, retrospective and observational study of IBD paediatric patients transferred to an adult IBD unit between 2017–2020. Two groups were compared: transition (≥1 joint visit involving the gastroenterologist, the paediatrician, a programme coordinator, the parents and the patient) and no-transition. Outcomes within one year after transfer were analysed. The main variable was poor clinical outcome (IBD flare, hospitalisation, surgery or any change in the treatment because of a flare). Predictive factors of poor clinical outcome were identified with multivariable analysis. (3) Results: A total of 278 patients from 34 Spanish hospitals were included. One hundred eighty-five patients (67%) from twenty-two hospitals (65%) performed a structured transition. Eighty-nine patients had poor clinical outcome at one year after transfer: 27% in the transition and 43% in the no-transition group (p = 0.005). One year after transfer, no-transition patients were more likely to have a flare (36% vs. 22%; p = 0.018) and reported more hospitalisations (10% vs. 3%; p = 0.025). The lack of transition, as well as parameters at transfer, including IBD activity, body mass index < 18.5 and corticosteroid treatment, were associated with poor clinical outcome. One patient in the transition group (0.4%) was lost to follow-up. (4) Conclusion: Transition care programmes improve patients’ outcomes after the transfer from paediatric to adult IBD units. Active IBD at transfer impairs outcomes., Depto. de Medicina, Depto. de Salud Pública y Materno - Infantil, Fac. de Medicina, TRUE, pub, Descuento UCM

This research was funded by Instituto de Salud Carlos III, grant number PI19/00792 (L.A.L.-F.) and Juan Rodes program JR19/00005 (E.L.-M.), by Instituto de Investigación Sanitaria Gregorio Marañón, grant number 2021-II-postdoc-01 (S.S.-M.), and by Consejería de Educación, Universidades, Ciencia y Portavocía Comunidad de Madrid, grant number PEJ-2021-AI/BMD-21866 (P.Z.-C.). The study was co-funded by the European Union., The genetic polymorphisms rs2395185 and rs2097432 in HLA genes have been associated with the response to anti-TNF treatment in inflammatory bowel disease (IBD). The aim was to analyze the association between these variants and the long-term response to anti-TNF drugs in pediatric IBD. We performed an observational, multicenter, ambispective study in which we selected 340 IBD patients under 18 years of age diagnosed with IBD and treated with anti-TNF drugs from a network of Spanish hospitals. Genotypes and failure of anti-TNF drugs were analyzed using Kaplan-Meier curves and Cox logistic regression. The homozygous G allele of rs2395185 and the C allele of rs2097432 were associated with impaired long-term response to anti-TNF drugs in children with IBD after 3 and 9 years of follow-up. Being a carrier of both polymorphisms increased the risk of anti-TNF failure. The SNP rs2395185 but not rs2097432 was associated with response to infliximab in adults with CD treated with infliximab but not in children after 3 or 9 years of follow-up. Conclusions: SNPs rs2395185 and rs2097432 were associated with a long-term response to anti-TNFs in IBD in Spanish children. Differences between adults and children were observed in patients diagnosed with CD and treated with infliximab., European Union, Consejería de Educación, Universidades, Ciencia y Portavocía Comunidad de Madrid, Instituto de Salud Carlos III, Juan Rodes program, Instituto de Investigación Sanitaria Gregorio Marañón, Depto. de Medicina, Fac. de Medicina, TRUE, pub

Comisión Europea, Depto. de Nutrición y Ciencia de los Alimentos, Fac. de Farmacia, FALSE, pub, Descuento UCM

This article aims to review the work of the German anatomist Joseph Disse (1852-1912), specifically with regard to the contents of his ,, Ueber die Lymphbahnen der Laugethierleber". In this he described a thin space, until that moment not referred, located between the hepatocyte and the sinusoidal membrane (sinusoids)., Depto. de Anatomía y Embriología, Depto. de Cirugía, Fac. de Medicina, TRUE, pub

Aims: Children with HIV exhibit chronic inflammation and immune dysfunction despite antiretroviral therapy (ART). Strategies targeting persistent inflammation are needed to improve health in people living with HIV. The gut microbiota likely interacts with the immune system, but the clinical implications of modulating the dysbiosis by nutritional supplementation are unclear. Methods: Pilot, double-blind, randomized placebo-controlled trial in which 24 HIV-infected on ART were randomized to supplementation with a daily mixture of symbiotics, omega-3/6 fatty acids and amino acids, or placebo four weeks, in combination with ART. We analyzed inflammatory markers and T-cell activation changes and their correlations with shifts in fecal microbiota. Results: Twenty-four HIV-infected children were recruited and randomized to receive a symbiotic nutritional supplement or placebo. Mean age was 12 ± 3.9 years, 62.5% were female. All were on ART and had HIV RNA < 50/mL. We did not detect changes in inflammatory (IL-6, IL-7, IP-10), microbial translocation (sCD14), mucosal integrity markers (IFABP, zonulin) or the kynurenine to tryptophan ratio, or changes in markers of the adaptive immune response in relation to the intervention. However, we found correlations between several key bacteria and the assessed inflammatory and immunological parameters, supporting a role of the microbiota in immune modulation in children with HIV. Conclusions: In this exploratory study, a four-week nutritional supplementation had no significant effects in terms of decreasing inflammation, microbial translocation, or T-cell activation in HIV-infected children. However, the correlations found support the interaction between gut microbiota and the immune system., Instituto de Salud Carlos III (ISCIII)/ FEDER, Instituto de Salud Carlos III, Spanish Society of Microbiology and Infectious Diseases (SEIMC)/Spanish Ministry of Science and Innovation/Instituto de Salud Carlos III/FEDER, Depto. de Medicina, Depto. de Salud Pública y Materno - Infantil, Fac. de Medicina, TRUE, pub

Nowadays, the food industry has developed novel gluten free extruded snack type products made from pulses, which could be good candidates to promote pulse consumption as a sustainable food product, while also satisfying the consumer’s demand. Snack type products are a large part of the young people’s diets and impact health outcomes, so it is essential to offer them snacks with a better nutritional profile. In this study, 81 Spanish millennial consumers tasted “in situ” six different gluten free snacks based on pulse flour (lentil and chickpea) marketed in Spain. The aim of the present study was: (a) to evaluate the Spanish millennial consumers’ acceptance level of new pulse snack type products; (b) to evaluate the segmentation of the millennial consumers and understand the difference between the segments; (c) to evaluate the potential relationship between their nutrition food labelling and consumers’ acceptance. In general, the lentil formulations (with more protein, more fat and less fiber) obtained higher scores than those of the chickpea. In addition, a multidimensional statistical analysis, preference mapping, and a statistical analysis of agglomerative hierarchical clustering were performed. Consumers were grouped into three clusters based on their preferences, allowing a detailed study of consumer acceptance of the selected snacks. Cluster 1 like less the samples with less salt, and, on the contrary, these samples were preferred by Cluster 3. Cluster 2 is a group who like lentil snacks, regardless of their flavoring. It was observed that the consumer segments differ at least in their preference for saltiness. The findings of this study also showed that the nutritional composition of the analyzed snacks (as appears in nutrition labelling) was associated with Spanish millennial consumers’ acceptance and could provide valuable information to develop new snacks targeted at specific market niches, such as millennials. These data provide valuable insights when tryi, Universidad Complutense de Madrid, Depto. de Nutrición y Ciencia de los Alimentos, Fac. de Farmacia, TRUE, pub

Artificial sweeteners are additives widely used in our diet. Although there is no consensus, current evidence indicates that sucralose and saccharin could influence the gut microbiota. The aim of this study was to analyze the existing scientific evidence on the effects of saccharin and sucralose consumption on gut microbiota in humans. Different databases were used with the following search terms: sweeteners, non-caloric-sweeteners, sucralose, splenda, saccharin, sugartwin, sweet’n low, microbiota, gut microbiota, humans, animal model, mice, rats, and/or in vitro studies. In vitro and animal model studies indicate a dose-dependent relationship between the intake of both sweeteners and gut microbiota affecting both diversity and composition. In humans, long-term study suggests the existence of a positive correlation between sweetener consumption and some bacterial groups; however, most short-term interventions with saccharin and sucralose, in amounts below the ADI, found no significant effect on those groups, but there seems to be a different basal microbiota-dependent response of metabolic markers. Although studies in vitro and in animal models seem to relate saccharin and sucralose consumption to changes in the gut microbiota, more long-term studies are needed in humans considering the basal microbiota of participants and their dietary and lifestyle habits in all population groups. Toxicological and basal gut microbiota effects must be included as relevant factors to evaluate food safety and nutritional consequences of non-calorie sweeteners. In humans, doses, duration of interventions, and number of subjects included in the studies are key factors to interpret the results., Depto. de Genética, Fisiología y Microbiología, Depto. de Nutrición y Ciencia de los Alimentos, Fac. de Ciencias Biológicas, Fac. de Farmacia, TRUE, pub

CRUE-CSIC (Acuerdos Transformativos 2022), Diagnosis of sinusoidal obstruction syndrome (SOS) after hematopoietic cell transplantation (HCT) is based on clinical criteria including weight gain, ascites, hepatomegaly, and jaundice.[1] However, clinical and histological features and prognosis of SOS after liver transplantation (LT) seem to differ from SOS after HCT.[2, 3] We aimed to determine the characteristics and outcomes of SOS after LT., Depto. de Medicina, Fac. de Medicina, TRUE, pub

CRUE-CSIC (Acuerdos Transformativos 2022), Objective non-functioning gastrin-producing Neuroendocrine Neoplasms (NEN) of the duodenum are rare tumors of the gastrointestinal tract without a clinical syndrome due to gastrin-production. Their incidence has significantly increased in the last few years especially as an incidental finding during endoscopic studies. The aim of this study is to describe the characteristics of this emergent neoplasm, to provide more information on this rare pathology and its possible prognostic factors. Methods we performed a retrospective observational study based on the duodenal-NENs samples with positive staining for gastrin, registered in the Pathology Department of University Hospital 12-de-Octubre (Madrid, Spain) between 2000 and 2017. We excluded all those clinically functional [(Zollinger-Ellison-Syndrome) and/or gastrinemia >1000pg/ml], with a previous diagnosis of multiple endocrine neoplasia(MEN-syndrome) or a synchronous neoplasia. Clinicopathological and therapeutic variables, follow-up time, recurrence and mortality data were collected. Results 21 patients were included. Most of the tumors were diagnosed incidentally as a single small polypoid lesion, they were limited to mucosa/submucosa and had a low histological-grade. 4 patients (19%) presented metastatic involvement at diagnosis (lymphatic and/or hepatic). These 4 patients also had a high or intermediate mitotic grade and infiltration further than submucosa. Local resection was used in most cases as curative treatment. The median follow-up was 25 months. There were 2 relapses and 2 deaths attributed to the tumor with a 5-year disease-free-survival of 81%. Conclusions The majority of these tumors were diagnosed at a local stage and had a good prognosis with local treatment. Nevertheless, given the potential metastatic risk, a close follow-up and extensive study is necessary, especially in those with aggressive pathological factors such as deep infiltration or high-histological-grade., Depto. de Medicina, Depto. de Medicina Legal, Psiquiatría y Patología, Fac. de Medicina, TRUE, pub

CRUE-CSIC (Acuerdos Transformativos 2022), Embolization with microspheres is a therapeutic strategy based on the selective occlusion of the blood vessels feeding a tumor. This procedure is intraarterially performed in the clinical setting for the treatment of liver cancer. The practice has evolved over the last decade through the incorporation of drug loading ability, biodegradability and imageability with the subsequent added functionality for the physicians and improved clinical outcomes for the patients. This review highlights the evolution of the embolization systems developed through the analysis of the marketed embolic microspheres for the treatment of malignant hepatocellular carcinoma, namely the most predominant form of liver cancer. Embolic microspheres for the distinct modalities of embolization (i.e., bland embolization, chemoembolization and radioembolization) are here comprehensively compiled with emphasis on material characteristics and their impact on microsphere performance. Moreover, the future application of the embolics under clinical investigation is discussed along with the scientific and regulatory challenges ahead in the field., Ministerio de Ciencia e Innovación (MICINN), Universidad Complutense de Madrid/Banco de Santander, Depto. de Farmacia Galénica y Tecnología Alimentaria, Fac. de Farmacia, TRUE, pub

CRUE-CSIC (Acuerdos Transformativos 2022), Schwannomas are peripheral nerve sheath tumors. Due to their low incidence, few cases of colorectal schwannomas have been published, which increases the diagnostic challenge. The aim of this case report is to discuss the role of transvaginal ultrasound in different areas than the gynecological disorders, when on hands of properly trained professionals that perform systematized procedures. A 56-year-old woman consulted for postmenopausal genital bleeding. During transvaginal ultrasound, a colonic solid, hypervascularized mass of 23 × 26 mm was visualized. As a result of this incidental finding, the patient underwent a sigmoidectomy, with a final diagnosis of intestinal schwannoma. Transvaginal ultrasound is today one of the most useful and accurate diagnostic tools in the assessment of gynecological disorders. However,the proximity of other pelvic structures makes it possible to evaluate the presence of nongynecological conditions. This fact should encourage gynecologists to systematize the transvaginal ultrasound procedure., Depto. de Salud Pública y Materno - Infantil, Fac. de Medicina, TRUE, pub

Amyloidoses are characterized by the accumulation and aggregation of misfolded proteins into fibrils in different organs, leading to cell death and consequent organ dysfunction. The specific substitution of Leu 75 for Pro in Apolipoprotein A-I protein sequence (ApoA-I; L75P-ApoA-I) results in late onset amyloidosis, where deposition of extracellular protein aggregates damages the normal functions of the liver. In this work, we describe that the autophagic process is inhibited in the presence of the L75P-ApoA-I amyloidogenic variant in stably transfected human hepatocyte carcinoma cells. The L75P-ApoA-I amyloidogenic variant alters the redox status of the cells, resulting into excessive mitochondrial stress and consequent cell death. Moreover, L75P-ApoA-I induces an impairment of the autophagic flux. Pharmacological induction of autophagy or transfection-enforced overexpression of the pro-autophagic transcription factor EB (TFEB) restores proficient proteostasis and reduces oxidative stress in these experimental settings, suggesting that pharmacological stimulation of autophagy could be a promising target to alleviate ApoA-I amyloidosis., Depto. de Fisiología, Fac. de Medicina, TRUE, pub

CRUE-CSIC (Acuerdos Transformativos 2022), Background Single-anastomosis duodeno-ileal bypass with sleeve gastrectomy (SADI-S) is a simplification of the duodenal switch (DS) in which the alimentary limb is eliminated, and the common channel is lengthened from 200 to 300 cm. Short-term results have demonstrated that SADI-S is safe and reproducible and that weight loss and comorbidities resolution are comparable to biliopancreatic diversion or DS. Objective To analyze the long-term outcomes of SADI-S. Methods From May 2007 to December 2015, 164 patients were consecutively submitted to a one-step SADI-S. The mean age was 47 years, and the mean body mass index (BMI) was 45.8 kg/m2. A total of 101 patients had type 2 diabetes, 91 arterial hypertension, 81 obstructive apnea, and 118 dyslipidemia. Limb length was 200 cm in 50 cases, 250 cm in 99, and 300 cm in 15. Results There was no mortality. One patient had a gastric leak, and 2 patients had an anastomotic leak. A total of 25% of the patients were lost to follow-up at 10 years. Excess weight loss and total weight loss were 87% and 38% at 5 years and 80% and 34% at 10 years. A total of 12 patients were submitted to revisional surgery for hypoproteinemia. Preoperatively 41 diabetics were under insulin treatment; at 5 years, 7 remained with insulin and 12 at 10 years. Mean glycemia was 104 mg/dL at 5 years and 118 mg/dL at 10 years. Mean HbA1c was 5.51% at 5 years and 5.86 at 10 years. Conclusion In the long term, SADI-S offers satisfactory weight loss and comorbidities resolution., Depto. de Medicina, Fac. de Medicina, TRUE, pub

La infección por virus de la hepatitis C (VHC) es una de las principales comorbilidades en pacientes infectados por virus de la inmunodeficiencia humana (VIH) en España. Sin tratamiento da lugar a cirrosis y sus complicaciones en un número muy importante de pacientes, especialmente en la era en que el tratamiento antirretroviral de gran efectividad ha permitido evitar las complicaciones por el síndrome de inmunodeficiencia adquirida y prolongar la supervivencia de los pacientes infectados por el VIH. La erradicación de la infección crónica por el VHC evita la progresión de la hepatopatía y la morbimortalidad asociada, tanto en pacientes monoinfectados por VHC como en coinfectados por VIH y VHC. Hasta el año 2014 el tratamiento de la infección crónica por VHC consistía en pautas basadas en interferón (IFN). Este tratamiento tenía una efectividad limitada y alta toxicidad y en pacientes infectados por VIH su efectividad era aún menor. En los últimos años ha habido un gran avance en los tratamientos de la infección crónica por VHC basado en el desarrollo de los antivirales de acción directa (AAD) que han permitido diseñar pautas libres de IFN con una alta eficacia y seguridad en los ensayos clínicos pivotales. En estos estudios, aunque hubo algunos específicamente diseñados en población con VIH y VHC, la representación de pacientes coinfectados ha sido limitada y con pacientes muy seleccionados por lo que es difícil extrapolar los resultados a la población general con coinfección VIH/VHC...

Entre las principales complicaciones asociadas a la obesidad destacan la resistencia a insulina y la alteración del metabolismo glucídico y lipídico. Esta situación puede llevar consigo el desarrollo de comorbilidades como, por ejemplo, la enfermedad de hígado graso no alcohólico (EHGNA) o la diabetes mellitus tipo 2 (DM2), que presentan características comunes. La EHGNA es la patología hepática más frecuente en los países de Occidente, cuya prevalencia se encuentra en continuo crecimiento. Esta manifestación se define principalmente por la presencia de esteatosis en más del 5% de los hepatocitos y su progresión puede causar esteatohepatitis no alcohólica (EHNA). Aunque hay factores implicados en la progresión de esteatosis a EHNA que están bien caracterizados, como lipotoxicidad, estrés oxidativo y activación del sistema inmune, muchos otros todavía no se conocen. Debido a la falta de tratamiento de esta enfermedad, es necesario profundizar en el conocimiento de nuevos mediadores, como los microRNAs (miRNAs),y abordajes terapéuticos, como la terapia génica, para impedir el desarrollo y la progresión de la EHGNA...

El dolor abdominal agudo (DAA) es uno de los motivos de consulta más frecuentes en los servicios de urgencias hospitalarios (SUH). La apendicitis aguda (AA) es uno de los principales diagnósticos diferenciales a tener en cuenta en la edad pediátrica y en los adolescentes, siendo la causa más frecuente de cirugía urgente en dicho grupo etario. A pesar de los avances, el diagnóstico sigue siendo difícil, especialmente en las edades más tempranas. La utilidad de la historia clínica y la exploración física puede ser menor en comparación con la población adulta ya que los niños se asocian a una mayor frecuencia de presentación clínica atípica. Se han publicado diversas escalas clínico-analíticas como herramientas de ayuda para el proceso diagnóstico de los pacientes con sospecha de AA, reduciendo el tiempo necesario para el diagnóstico, el número de pruebas de imagen y apendicectomías inadecuadas. La escala de Alvarado (EA), la más utilizada hasta el momento, combina síntomas y signos conjuntamente con datos analíticos como la leucocitosis y la neutrofilia y, en función de la puntuación, da recomendaciones sobre el alta, la observación y la necesidad de intervención quirúrgica. Sin embargo, la precisión de estas escalas en la población pediátrica no está bien evaluada. En la actualidad, también se están desarrollando líneas de investigación basadas en nuevos biomarcadores como el APPY1 Test, que es una aproximación multimarcador a pie de cama que podría servir para descartar AA de una forma rápida y segura...

Los pacientes infectados perinatalmente por el virus de la inmunodeficiencia humana (PVIH) tienen una larga historia de exposición al virus y al tratamiento antirretroviral (TAR) desde una edad muy temprana, incluso algunos de manera intrauterina. Los efectos del virus y su tratamiento sobre un sistema inmune inmaduro podrían producir alteraciones permanentes en la respuesta inmune. En adultos que viven con el VIH, a pesar del TAR y el control de la viremia, la infección crónica conduce a un estado proinflamatorio persistente que desdencadena un proceso de envejecimiento acelerado y aumento de comorbilidades. Estas alteraciones inmunológicas podrían ser más acusadas en el caso de los pacientes infectados por vía perinatal. Las primeras evidencias sugieren que los PVIH presentan una mayor prevalencia de comorbilidades, cuando se comparan con individuos no infectados de su misma edad. Entre las comorbilidades más comunes asociadas a la infección están las enfermedades cardiovasculares, los tumores, las alteraciones metabólicas, las alteraciones neurocognitivas, la enfermedad renal y la enfermedad hepática. Esta memoria se centra en la enfermedad hepática en el contexto de la infección por el VIH de transmisión perinatal, en relación con la coinfección por el virus de la hepatitis C (VHC), así como en el contexto de la enfermedad metábolica y los potenciales mecanismos subyacentes en relación con el sistema inmunológico...

La creciente incidencia de la obesidad y la enfermedad del hígado graso no alcohólico (EHGNA/NASH) representa una seria amenaza para la salud mundial, pudiendo conducir a un aumento en el número de casos de enfermedad hepática avanzada, cirrosis y en última instancia derivar en carcinoma hepatocelular (CHC). El gen p21/CDKN1A es un regulador del ciclo celular que está involucrado en procesos celulares esenciales tales como: la detención del ciclo celular, la apoptosis, la diferenciación, la senescencia y la reparación del ADN. El objetivo principal de este estudio fue determinar el papel específico de p21/CDKN1A en la enfermedad hepática crónica...