RESUMEN
BACKGROUND AND AIMS: Alcohol-associated liver disease (ALD) accounts for 70% of liver-related deaths in Europe, with no effective approved therapies. Although mitochondrial dysfunction is one of the earliest manifestations of alcohol-induced injury, restoring mitochondrial activity remains a problematic strategy due to oxidative stress. Here, we identify methylation-controlled J protein (MCJ) as a mediator for ALD progression and hypothesize that targeting MCJ may help in recovering mitochondrial fitness without collateral oxidative damage. APPROACH AND RESULTS: C57BL/6 mice [wild-type (Wt)] Mcj knockout and Mcj liver-specific silencing (MCJ-LSS) underwent the NIAAA dietary protocol (Lieber-DeCarli diet containing 5% (vol/vol) ethanol for 10 days, plus a single binge ethanol feeding at day 11). To evaluate the impact of a restored mitochondrial activity in ALD, the liver, gut, and pancreas were characterized, focusing on lipid metabolism, glucose homeostasis, intestinal permeability, and microbiota composition. MCJ, a protein acting as an endogenous negative regulator of mitochondrial respiration, is downregulated in the early stages of ALD and increases with the severity of the disease. Whole-body deficiency of MCJ is detrimental during ALD because it exacerbates the systemic effects of alcohol abuse through altered intestinal permeability, increased endotoxemia, and dysregulation of pancreatic function, which overall worsens liver injury. On the other hand, liver-specific Mcj silencing prevents main ALD hallmarks, that is, mitochondrial dysfunction, steatosis, inflammation, and oxidative stress, as it restores the NAD + /NADH ratio and SIRT1 function, hence preventing de novo lipogenesis and improving lipid oxidation. CONCLUSIONS: Improving mitochondrial respiration by liver-specific Mcj silencing might become a novel therapeutic approach for treating ALD.
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Hepatopatías Alcohólicas , Animales , Ratones , Ratones Endogámicos C57BL , Hepatopatías Alcohólicas/metabolismo , Hígado/metabolismo , Etanol/efectos adversos , Mitocondrias/metabolismo , Chaperonas Moleculares/metabolismo , Proteínas Mitocondriales/metabolismoRESUMEN
Metabolic-associated steatotic liver disease (MASLD) encompasses a wide spectrum of metabolic conditions associated with an excess of fat accumulation in the liver, ranging from simple hepatic steatosis to cirrhosis and hepatocellular carcinoma. Finding appropriate tools to study its development and progression is essential to address essential unmet therapeutic and staging needs. This review discusses advantages and shortcomings of different dietary, chemical and genetic factors that can be used to mimic this disease and its progression in mice from a hepatic and metabolic point of view. Also, this review will highlight some additional factors and considerations that could have a strong impact on the outcomes of our model to end up providing recommendations and a checklist to facilitate the selection of the appropriate MASLD preclinical model based on clinical aims.
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Carcinoma Hepatocelular , Hígado Graso , Neoplasias Hepáticas , Animales , Ratones , Cirrosis HepáticaRESUMEN
OBJECTIVES: Fibromyalgia (FM) is a disease treated with various therapeutic approaches that have limited success. Pulsed electromagnetic field therapy has been proposed as a possible solution to reduce several symptoms. This study aims to analyse the therapeutic effects of transcranial low-intensity magnetic stimulation (LIMS) in women diagnosed with FM at 2, 12 and 24 weeks from the last LIMS administration treatment session. METHODS: 560 women (53.7 ± 11.3 years) diagnosed with FM according to the ACR 2016 criteria were randomly allocated in two groups: 280 received standard pharmacological treatment and 280 received the same treatment plus eight sessions of LIMS, 20 minutes long, once a week. The variables analysed were the widespread pain index (WPI), symptoms severity score (SS score) and the Spanish-validated version of the FM impact questionnaire (S-FIQ). The evaluations were performed at the beginning of LIMS treatment and at 2, 12 and 24 weeks after the end of the last LIMS treatment session. RESULTS: From the second week after the last LIMS session, there was significant improvement (p <0.001) in the variables WPI, SS score and S-FIQ. This improvement was maintained throughout the 24 weeks of monitoring after the last intervention. The age of the patients and the severity of the symptoms at the time of diagnosis did not affect the improvement observed in the three variables studied. CONCLUSIONS: Treatment with LIMS for eight weeks resulted in significant improvement in FM diagnostic variables, which was maintained up to 24 weeks after the last treatment session. This therapy could be recommended as a part of a multimodal approach for FM treatment.
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Fibromialgia , Femenino , Fibromialgia/diagnóstico , Fibromialgia/terapia , Humanos , Dolor , Dimensión del Dolor/métodos , Calidad de Vida , Índice de Severidad de la Enfermedad , Encuestas y Cuestionarios , Estimulación Magnética TranscranealRESUMEN
The transcription factor, early growth response-1 (EGR-1), is involved in the regulation of cell differentiation, proliferation, and apoptosis in response to different stimuli. EGR-1 is described to be involved in pancreatic endoderm differentiation, but the regulatory mechanisms controlling its action are not fully elucidated. Our previous investigation reported that exposure of mouse embryonic stem cells (mESCs) to the chemical nitric oxide (NO) donor diethylenetriamine nitric oxide adduct (DETA-NO) induces the expression of early differentiation genes such as pancreatic and duodenal homeobox 1 (Pdx1). We have also evidenced that Pdx1 expression is associated with the release of polycomb repressive complex 2 (PRC2) and P300 from the Pdx1 promoter; these events were accompanied by epigenetic changes to histones and site-specific changes in the DNA methylation. Here, we investigate the role of EGR-1 on Pdx1 regulation in mESCs. This study reveals that EGR-1 plays a negative role in Pdx1 expression and shows that the binding capacity of EGR-1 to the Pdx1 promoter depends on the methylation level of its DNA binding site and its acetylation state. These results suggest that targeting EGR-1 at early differentiation stages might be relevant for directing pluripotent cells into Pdx1-dependent cell lineages.
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Endodermo , Células Madre Embrionarias de Ratones , Animales , Diferenciación Celular/genética , Células Madre Embrionarias , Endodermo/metabolismo , Ratones , Óxido Nítrico/metabolismoRESUMEN
This is a meeting report of the 3rd Translational Hepatology Meeting held in Alicante, Spain, in October 2021. The meeting, which was organized by the Spanish Association for the Study of the Liver (AEEH), provided an update on the recent advances in the field of basic and translational hepatology, with a particular focus on the molecular and cellular mechanisms and therapeutic targets involved in metabolic-associated fatty liver disease (MAFLD), metabolic-associated steatohepatitis (MASH), cirrhosis and end-stage hepatocellular carcinoma (HCC).
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Carcinoma Hepatocelular , Gastroenterología , Neoplasias Hepáticas , Enfermedad del Hígado Graso no Alcohólico , Humanos , Carcinoma Hepatocelular/terapia , Carcinoma Hepatocelular/complicaciones , Neoplasias Hepáticas/terapia , Neoplasias Hepáticas/complicaciones , Enfermedad del Hígado Graso no Alcohólico/complicaciones , Enfermedad del Hígado Graso no Alcohólico/terapia , Enfermedad del Hígado Graso no Alcohólico/patología , Cirrosis Hepática/complicaciones , Cirrosis Hepática/patologíaRESUMEN
BACKGROUND & AIMS: Perturbations of intracellular magnesium (Mg2+) homeostasis have implications for cell physiology. The cyclin M family, CNNM, perform key functions in the transport of Mg2+ across cell membranes. Herein, we aimed to elucidate the role of CNNM4 in the development of non-alcoholic steatohepatitis (NASH). METHODS: Serum Mg2+ levels and hepatic CNNM4 expression were characterised in clinical samples. Primary hepatocytes were cultured under methionine and choline deprivation. A 0.1% methionine and choline-deficient diet, or a choline-deficient high-fat diet were used to induce NASH in our in vivo rodent models. Cnnm4 was silenced using siRNA, in vitro with DharmaFECT and in vivo with Invivofectamine® or conjugated to N-acetylgalactosamine. RESULTS: Patients with NASH showed hepatic CNNM4 overexpression and dysregulated Mg2+ levels in the serum. Cnnm4 silencing ameliorated hepatic lipid accumulation, inflammation and fibrosis in the rodent NASH models. Mechanistically, CNNM4 knockdown in hepatocytes induced cellular Mg2+ accumulation, reduced endoplasmic reticulum stress, and increased microsomal triglyceride transfer activity, which promoted hepatic lipid clearance by increasing the secretion of VLDLs. CONCLUSIONS: CNNM4 is overexpressed in patients with NASH and is responsible for dysregulated Mg2+ transport. Hepatic CNNM4 is a promising therapeutic target for the treatment of NASH. LAY SUMMARY: Cyclin M4 (CNNM4) is overexpressed in non-alcoholic steatohepatitis (NASH) and promotes the export of magnesium from the liver. The liver-specific silencing of Cnnm4 ameliorates NASH by reducing endoplasmic reticulum stress and promoting the activity of microsomal triglyceride transfer protein.
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Proteínas Portadoras/metabolismo , Proteínas de Transporte de Catión/metabolismo , Hepatocitos/metabolismo , Magnesio , Enfermedad del Hígado Graso no Alcohólico , Animales , Transporte Biológico/efectos de los fármacos , Células Cultivadas , Modelos Animales de Enfermedad , Descubrimiento de Drogas , Estrés del Retículo Endoplásmico/efectos de los fármacos , Regulación de la Expresión Génica , Humanos , Magnesio/sangre , Magnesio/metabolismo , Ratones , Enfermedad del Hígado Graso no Alcohólico/metabolismo , Enfermedad del Hígado Graso no Alcohólico/patologíaRESUMEN
Pancreatic heterotopia is defined as pancreatic tissue outside its normal location in the body and anatomically separated from the pancreas. In this work we have analyzed the stomach glandular epithelium of Gata4 flox/flox ; Pdx1-Cre mice (Gata4KO mice). We found that Gata4KO glandular epithelium displays an atypical morphology similar to the cornified squamous epithelium and exhibits upregulation of forestomach markers. The developing gastric units fail to form properly, and the glandular epithelial cells do not express markers of gastric gland in the absence of GATA4. Of interest, the developing glands of the Gata4KO stomach express pancreatic cell markers. Furthermore, a mass of pancreatic tissue located in the subserosa of the Gata4KO stomach is observed at adult stages. Heterotopic pancreas found in Gata4-deficient mice contains all three pancreatic cell lineages: ductal, acinar, and endocrine. Moreover, Gata4 expression is downregulated in ectopic pancreatic tissue of some human biopsy samples. © 2019 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
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Células Epiteliales/patología , Factor de Transcripción GATA4/genética , Páncreas/patología , Animales , Diferenciación Celular/genética , Diferenciación Celular/fisiología , Mucosa Gástrica/metabolismo , Regulación del Desarrollo de la Expresión Génica/genética , Humanos , Ratones Transgénicos , Organogénesis/fisiologíaRESUMEN
The secondary reconstruction of flexor tendons is in most cases very demanding and tedious. The indications, selection of the correct surgical procedure, operative technique and further treatment have to be individually adjusted and are mostly very difficult. Due to the previous operations unpleasant surprises may occur intraoperatively, which must be recognized and treated by the surgeon. Nevertheless, a significant improvement of the function of the whole hand can be achieved for most patients, e.g. by a two-stage flexor tendon transplantation or other techniques described in this article.
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Traumatismos de los Dedos , Traumatismos de la Mano , Procedimientos Ortopédicos , Traumatismos de los Tendones , Traumatismos de los Dedos/cirugía , Mano , Traumatismos de la Mano/cirugía , Humanos , Traumatismos de los Tendones/cirugíaRESUMEN
AIMS/HYPOTHESIS: Bisphenol-A (BPA) is a widespread endocrine-disrupting chemical that has been associated with type 2 diabetes development. Low doses of BPA modify pancreatic beta cell function and induce insulin resistance; some of these effects are mediated via activation of oestrogen receptors α (ERα) and ß (ERß). Here we investigated whether low doses of BPA regulate the expression and function of ion channel subunits involved in beta cell function. METHODS: Microarray gene profiling of isolated islets from vehicle- and BPA-treated (100 µg/kg per day for 4 days) mice was performed using Affymetrix GeneChip Mouse Genome 430.2 Array. Expression level analysis was performed using the normalisation method based on the processing algorithm 'robust multi-array average'. Whole islets or dispersed islets from C57BL/6J or oestrogen receptor ß (ERß) knockout (Erß-/-) mice were treated with vehicle or BPA (1 nmol/l) for 48 h. Whole-cell patch-clamp recordings were used to measure Na+ and K+ currents. mRNA expression was evaluated by quantitative real-time PCR. RESULTS: Microarray analysis showed that BPA modulated the expression of 1440 probe sets (1192 upregulated and 248 downregulated genes). Of these, more than 50 genes, including Scn9a, Kcnb2, Kcnma1 and Kcnip1, encoded important Na+ and K+ channel subunits. These findings were confirmed by quantitative RT-PCR in islets from C57BL/6J BPA-treated mice or whole islets treated ex vivo. Electrophysiological measurements showed a decrease in both Na+ and K+ currents in BPA-treated islets. The pharmacological profile indicated that BPA reduced currents mediated by voltage-activated K+ channels (Kv2.1/2.2 channels) and large-conductance Ca2+-activated K+ channels (KCa1.1 channels), which agrees with BPA's effects on gene expression. Beta cells from ERß-/- mice did not present BPA-induced changes, suggesting that ERß mediates BPA's effects in pancreatic islets. Finally, BPA increased burst duration, reduced the amplitude of the action potential and enlarged the action potential half-width, leading to alteration in beta cell electrical activity. CONCLUSIONS/INTERPRETATION: Our data suggest that BPA modulates the expression and function of Na+ and K+ channels via ERß in mouse pancreatic islets. Furthermore, BPA alters beta cell electrical activity. Altogether, these BPA-induced changes in beta cells might play a role in the diabetogenic action of BPA described in animal models.
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Compuestos de Bencidrilo/farmacología , Diabetes Mellitus Tipo 2/metabolismo , Receptor beta de Estrógeno/metabolismo , Células Secretoras de Insulina/efectos de los fármacos , Células Secretoras de Insulina/metabolismo , Fenoles/farmacología , Animales , Receptor alfa de Estrógeno/metabolismo , Islotes Pancreáticos/efectos de los fármacos , Islotes Pancreáticos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Potasio/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Sodio/metabolismoRESUMEN
Diabetes is a chronic metabolic disorder that affects 415 million people worldwide. This pathology is often associated with long-term complications, such as critical limb ischemia (CLI), which increases the risk of limb loss and mortality. Mesenchymal stromal cells (MSCs) represent a promising option for the treatment of diabetes complications. Although MSCs are widely used in autologous cell-based therapy, their effects may be influenced by the constant crosstalk between the graft and the host, which could affect the MSC fate potential. In this context, we previously reported that MSCs derived from diabetic patients with CLI have a defective phenotype that manifests as reduced fibrinolytic activity, thereby enhancing the thrombotic risk and compromising patient safety. Here, we found that MSCs derived from diabetic patients with CLI not only exhibit a prothrombotic profile but also have altered multi-differentiation potential, reduced proliferation, and inhibited migration and homing to sites of inflammation. We further demonstrated that this aberrant cell phenotype is reversed by the platelet-derived growth factor (PDGF) BB, indicating that PDGF signaling is a key regulator of MSC functionality. These findings provide an attractive approach to improve the therapeutic efficacy of MSCs in autologous therapy for diabetic patients.
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Diabetes Mellitus/genética , Inflamación/genética , Células Madre Mesenquimatosas/metabolismo , Proteínas Proto-Oncogénicas c-sis/genética , Animales , Diferenciación Celular/genética , Proliferación Celular/genética , Tratamiento Basado en Trasplante de Células y Tejidos/métodos , Células Cultivadas , Complicaciones de la Diabetes/genética , Complicaciones de la Diabetes/patología , Complicaciones de la Diabetes/terapia , Diabetes Mellitus/metabolismo , Diabetes Mellitus/patología , Diabetes Mellitus/terapia , Humanos , Inflamación/patología , Inflamación/terapia , Ratones , Ratones SCID , Osteogénesis/genética , Fenotipo , Proteínas Proto-Oncogénicas c-sis/uso terapéutico , Transducción de Señal , Cicatrización de Heridas/genéticaRESUMEN
BACKGROUND: Alcoholic fermentation of fruits has generated novel products with high concentrations of bioactive compounds and moderate alcohol content. The aim of this study was to evaluate the potential effect on cardiovascular risk factors of the regular consumption by healthy humans of a beverage obtained by alcoholic fermentation and pasteurization of orange juice. RESULTS: Thirty healthy volunteers were enrolled in a randomized controlled study. The experimental group (n = 15) drank 500 mL orange beverage (OB) per day for 2 weeks (intervention phase), followed by a 3-week washout phase. Blood samples were collected at baseline (E-T0) and at the end of the intervention (E-T1) and washout (E-T2) phases. Controls (n = 15) did not consume OB during a 2-week period. OB intake significantly increased oxygen radical absorbance capacity (43.9%) and reduced uric acid (-8.9%), catalase (CAT) (-23.2%), thiobarbituric acid reactive substances (TBARS) (-30.2%) and C-reactive protein (-2.1%) (E-T1 vs. E-T0). These effects may represent longer-term benefits, given the decreased uric acid (-8.9%), CAT (-34.6%), TBARS (-48.4%) and oxidized low-density lipoprotein (-23.9%) values recorded after the washout phase (E-T2 vs. E-T0). CONCLUSION: The regular consumption of OB improved antioxidant status and decreased inflammation state, lipid peroxidation and uric acid levels. Thus OB may protect the cardiovascular system in healthy humans and be considered a novel functional beverage. © 2017 Society of Chemical Industry.
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Antioxidantes/metabolismo , Bebidas/análisis , Citrus sinensis/metabolismo , Jugos de Frutas y Vegetales/microbiología , Inflamación/dietoterapia , Peroxidación de Lípido , Pichia/metabolismo , Adulto , Biomarcadores/metabolismo , Proteína C-Reactiva/metabolismo , Catalasa/metabolismo , Citrus sinensis/química , Citrus sinensis/microbiología , Femenino , Fermentación , Frutas/química , Frutas/metabolismo , Frutas/microbiología , Jugos de Frutas y Vegetales/análisis , Voluntarios Sanos , Humanos , Inflamación/genética , Inflamación/metabolismo , Lipoproteínas LDL/metabolismo , Masculino , Persona de Mediana Edad , Estrés Oxidativo , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismoRESUMEN
Efficient induction of defined lineages in pluripotent stem cells constitutes the determinant step for the generation of therapeutically relevant replacement cells to potentially treat a wide range of diseases, including diabetes. Pancreatic differentiation has remained an important challenge in large part because of the need to differentiate uncommitted pluripotent stem cells into highly specialized hormone-secreting cells, which has been shown to require a developmentally informed step-by-step induction procedure. Here, in the framework of using induced pluripotent stem cells (iPSCs) to generate pancreatic cells for pancreatic diseases, we have generated and characterized iPSCs from Pdx1-GFP transgenic mice. The use of a GFP reporter knocked into the endogenous Pdx1 promoter allowed us to monitor pancreatic induction based on the expression of Pdx1, a pancreatic master transcription factor, and to isolate a pure Pdx1-GFP+ population for downstream applications. Differentiated cultures timely expressed markers specific to each stage and end-stage progenies acquired a rather immature beta-cell phenotype, characterized by polyhormonal expression even among cells highly expressing the Pdx1-GFP reporter. Our findings highlight the utility of employing a fluorescent protein reporter under the control of a master developmental gene in order to devise novel differentiation protocols for relevant cell types for degenerative diseases such as pancreatic beta cells for diabetes.
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Diferenciación Celular/genética , Proteínas de Homeodominio/genética , Células Madre Pluripotentes Inducidas/citología , Páncreas/crecimiento & desarrollo , Transactivadores/genética , Animales , Células Madre Embrionarias/citología , Genes Reporteros , Proteínas Fluorescentes Verdes/genética , Ratones , Ratones Transgénicos , Páncreas/citología , Regiones Promotoras GenéticasRESUMEN
Consistent evidence from both experimental and human studies indicates that Type 2 diabetes mellitus (T2DM) is a complex disease resulting from the interaction of genetic, epigenetic, environmental, and lifestyle factors. Nutrients and dietary patterns are important environmental factors to consider in the prevention, development and treatment of this disease. Nutritional genomics focuses on the interaction between bioactive food components and the genome and includes studies of nutrigenetics, nutrigenomics and epigenetic modifications caused by nutrients. There is evidence supporting the existence of nutrient-gene and T2DM interactions coming from animal studies and family-based intervention studies. Moreover, many case-control, cohort, cross-sectional cohort studies and clinical trials have identified relationships between individual genetic load, diet and T2DM. Some of these studies were on a large scale. In addition, studies with animal models and human observational studies, in different countries over periods of time, support a causative relationship between adverse nutritional conditions during in utero development, persistent epigenetic changes and T2DM. This review provides comprehensive information on the current state of nutrient-gene interactions and their role in T2DM pathogenesis, the relationship between individual genetic load and diet, and the importance of epigenetic factors in influencing gene expression and defining the individual risk of T2DM.
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Diabetes Mellitus Tipo 2/genética , Dieta , Regulación de la Expresión Génica , Nutrigenómica , Animales , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/patología , Dieta/efectos adversos , Epigénesis Genética , Predisposición Genética a la Enfermedad , Variación Genética , Humanos , Células Secretoras de Insulina/metabolismo , Células Secretoras de Insulina/patología , Nutrigenómica/métodosRESUMEN
Nitric oxide (NO) delays mouse embryonic stem cell (mESC) differentiation by regulating genes linked to pluripotency and differentiation. Nevertheless, no profound study has been conducted on cell differentiation regulation by this molecule through signaling on essential biological functions. We sought to demonstrate that NO positively regulates the pluripotency transcriptional core, enforcing changes in the chromatin structure, in addition to regulating cell proliferation, and signaling pathways with key roles in stemness. Culturing mESCs with 2 µM of the NO donor diethylenetriamine/NO (DETA/NO) in the absence of leukemia inhibitory factor (LIF) induced significant changes in the expression of 16 genes of the pluripotency transcriptional core. Furthermore, treatment with DETA/NO resulted in a high occupancy of activating H3K4me3 at the Oct4 and Nanog promoters and repressive H3K9me3 and H3k27me3 at the Brachyury promoter. Additionally, the activation of signaling pathways involved in pluripotency, such as Gsk3-ß/ß-catenin, was observed, in addition to activation of PI3 K/Akt, which is consistent with the protection of mESCs from cell death. Finally, a decrease in cell proliferation coincides with cell cycle arrest in G2/M. Our results provide novel insights into NO-mediated gene regulation and cell proliferation and suggest that NO is necessary but not sufficient for the maintenance of pluripotency and the prevention of cell differentiation. J. Cell. Biochem. 117: 2078-2088, 2016. © 2016 Wiley Periodicals, Inc.
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Diferenciación Celular/fisiología , Puntos de Control de la Fase G2 del Ciclo Celular/fisiología , Expresión Génica/fisiología , Puntos de Control de la Fase M del Ciclo Celular/fisiología , Células Madre Embrionarias de Ratones/metabolismo , Óxido Nítrico/metabolismo , Transducción de Señal/fisiología , Animales , Diferenciación Celular/efectos de los fármacos , Línea Celular , Puntos de Control de la Fase G2 del Ciclo Celular/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Puntos de Control de la Fase M del Ciclo Celular/efectos de los fármacos , Ratones , Células Madre Embrionarias de Ratones/citología , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal/efectos de los fármacos , Triazenos/farmacologíaRESUMEN
Previously, we reported that alcoholic fermentation enhanced flavanones and carotenoids content of orange juice. The aim of this work was to evaluate the influence of pasteurization on the qualitative and quantitative profile of bioactive compounds and the antioxidant capacity of fermented orange juice. Ascorbic acid (203 mg/L), total flavanones (647 mg/L), total carotenoids (7.07 mg/L) and provitamin A (90.06 RAEs/L) values of pasteurized orange beverage were lower than those of fermented juice. Total phenolic remained unchanged (585 mg/L) and was similar to that of original juice. The flavanones naringenin-7-O-glucoside, naringenin-7-O-rutinoside, hesperetin-7-O-rutinoside, hesperetin-7-O-glucoside and isosakuranetin-7-O-rutinoside, and the carotenoids karpoxanthin and isomer, neochrome, lutein, ζ-carotene, zeaxanthin, mutatoxanthin epimers, ß-cryptoxanthin and auroxanthin epimers were the major compounds. Pasteurization produced a decrease in antioxidant capacity of fermented juice. However, TEAC (5.45 mM) and ORAC (6353 µM) values of orange beverage were similar to those of original orange juice. The novel orange beverage could be a valuable source of bioactive compounds with antioxidant capacity and exert potential beneficial effects.
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Antioxidantes/análisis , Citrus sinensis/química , Manipulación de Alimentos , Jugos de Frutas y Vegetales/análisis , Calor , Ácido Ascórbico/análisis , Carotenoides/análisis , Estudios de Evaluación como Asunto , Fermentación , Flavanonas/análisis , Flavonoides/análisis , Frutas/química , Glucósidos/análisis , Hesperidina/análogos & derivados , Hesperidina/análisis , Pasteurización , Fenoles/análisis , Provitaminas/análisis , Vitamina A/análisisRESUMEN
BACKGROUND & AIMS: Hepatocellular carcinoma (HCC) associated macrophages accelerate tumor progression by growth factor release. Therefore, tumor-associated macrophages (TAM) and their initiated signaling cascades are potential therapeutic targets. Aiming at understanding anticancer effects of systemic HCC therapy, we investigated the impact of sorafenib on macrophage function, focusing on macrophage-related growth factor secretion. METHODS: Macrophage markers, cytokine and growth factor release were investigated in CSF-1 (M1) or GMCSF (M2) maturated monocyte-derived macrophages. Macrophages were treated with sorafenib (1.2-5.0 µg/ml) and culture supernatants were transferred to hepatoma cell cultures to assess growth propagation. Insulin-like growth factor (IGF) signaling was blocked with NVP-AEW541 to confirm the role of IGF-1 in macrophage-driven hepatoma cell propagation. Macrophage activation was followed by ELISA of serum soluble mCD163 in sorafenib-treated patients with HCC. RESULTS: Alternative macrophages (M2), which showed higher IGF-1 (p=0.022) and CD163 mRNA (p=0.032) expression compared to classical macrophages (M1), increased hepatoma growth. This effect was mediated by M2-conditioned culture media. In turn, sorafenib lowered mCD163 and IGF-1 release by M2 macrophages, which decelerated M2 macrophage driven HuH7 and HepG2 proliferation by 47% and 64%, respectively. IGF-receptor blockage with NVP-AEW541 reduced growth induction by M2-conditioned culture media in a dose dependent manner. A transient mCD163 reduction during sorafenib treatment indicated a coherent M2 macrophage inhibition in patients with HCC. CONCLUSIONS: Sorafenib alters macrophage polarization, reduces IGF-1-driven cancer growth in vitro and partially inhibits macrophage activation in vivo. Thus macrophage modulation might contribute to the anti-cancer activity of sorafenib. However, more efficient macrophage-directed therapies are required.
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Carcinoma Hepatocelular , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Neoplasias Hepáticas , Factor Estimulante de Colonias de Macrófagos/metabolismo , Macrófagos , Niacinamida/análogos & derivados , Compuestos de Fenilurea/farmacología , Antígenos CD/metabolismo , Antígenos de Diferenciación Mielomonocítica/metabolismo , Antineoplásicos/farmacología , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patología , Línea Celular Tumoral , Humanos , Factor I del Crecimiento Similar a la Insulina/metabolismo , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , Activación de Macrófagos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Niacinamida/farmacología , Receptores de Superficie Celular/metabolismo , Receptores de Factores de Crecimiento Endotelial Vascular/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Sorafenib , Quinasas raf/metabolismoRESUMEN
Mouse embryonic stem cells (mESCs) are capable of both self-renewal and multilineage differentiation; thus, they can be expanded in vivo or in vitro and differentiated to produce different cell types. Despite their biological and medical interest, many physiological properties of undifferentiated mESCs, such as ion channel function, are not fully understood. Ion channels are thought to be involved in cell proliferation and differentiation. The aim of this study was to characterize functional ion channels in cultured undifferentiated mESCs and their role in cell proliferation. L-type voltage-activated Ca(2+) channels sensitive to nifedipine and small-conductance Ca(2+)-activated K(+) (SK) channels sensitive to apamin were identified. Ca(2+)-activated K(+) currents were blocked by millimolar concentrations of tetraethylammonium. The effects of Ca(2+) channel and Ca(2+)-activated K(+) channel blockers on the proliferation of undifferentiated mESCs were investigated by bromodeoxyuridine (BrdU) incorporation. Dihydropyridine derivatives, such as nifedipine, inhibited cell growth and BrdU incorporation into the cells, whereas apamin, which selectively blocks SK channels, had no effect on cell growth. These results demonstrate that functional voltage-operated Ca(2+) channels and Ca(2+)-activated K(+) channels are present in undifferentiated mESCs. Moreover, voltage-gated L-type Ca(2+) channels, but not SK channels, might be necessary for proliferation of undifferentiated mESCs.
Asunto(s)
Canales de Calcio Tipo L/metabolismo , Proliferación Celular/fisiología , Células Madre Embrionarias de Ratones/metabolismo , Canales de Potasio de Pequeña Conductancia Activados por el Calcio/metabolismo , Animales , Bloqueadores de los Canales de Calcio/farmacología , Línea Celular , Proliferación Celular/efectos de los fármacos , Transporte Iónico/efectos de los fármacos , Transporte Iónico/fisiología , Ratones , Células Madre Embrionarias de Ratones/citología , Canales de Potasio de Pequeña Conductancia Activados por el Calcio/antagonistas & inhibidores , Tetraetilamonio/farmacologíaRESUMEN
UNLABELLED: Alternatively polarized macrophages (MÏ) shape the microenvironment of hepatocellular carcinoma (HCC) and temper anticancer immune responses. We investigated if sorafenib alters the HCC microenvironment by restoring classical macrophage polarization and triggering tumor-directed natural killer (NK) cell responses. In vivo experiments were conducted with sorafenib (25 mg/kg)-treated C57BL/6 wildtype as well as hepatitis B virus (HBV) and lymphotoxin transgenic mice with and without HCC. Monocyte-derived MÏ or tumor-associated macrophages (TAM) isolated from HCC tissue were treated with sorafenib (0.07-5.0 µg/mL) and cocultured with autologous NK cells. MÏ and NK cell activation was analyzed by flow cytometry and killing assays, respectively. Cytokine and growth factor release was measured by enzyme-linked immunosorbent assay. Short-term administration of sorafenib triggered activation of hepatic NK cells in wildtype and tumor-bearing mice. In vitro, sorafenib sensitized MÏ to lipopolysaccharide, reverted alternative MÏ polarization and enhanced IL12 secretion (P = 0.0133). NK cells activated by sorafenib-treated MÏ showed increased degranulation (15.3 ± 0.2% versus 32.0 ± 0.9%, P < 0.0001) and interferon-gamma (IFN-γ) secretion (2.1 ± 0.2% versus 8.0 ± 0.2%, P < 0.0001) upon target cell contact. Sorafenib-triggered NK cell activation was verified by coculture experiments using TAM. Sorafenib-treated MÏ increased cytolytic NK cell function against K562, Raji, and HepG2 target cells in a dose-dependent manner. Neutralization of interleukin (IL)12 or IL18 as well as inhibition of the nuclear factor kappa B (NF-κB) pathway reversed NK cell activation in MÏ/NK cocultures. CONCLUSION: Sorafenib triggers proinflammatory activity of TAM and subsequently induces antitumor NK cell responses in a cytokine- and NF-κB-dependent fashion. This observation is relevant for HCC therapy, as sorafenib is a compound in clinical use that reverts alternative polarization of TAM in HCC. (HEPATOLOGY 2013;57:2358-2368).
Asunto(s)
Antineoplásicos/farmacología , Carcinoma Hepatocelular/tratamiento farmacológico , Neoplasias Hepáticas/tratamiento farmacológico , Activación de Macrófagos/efectos de los fármacos , Niacinamida/análogos & derivados , Compuestos de Fenilurea/farmacología , Animales , Antineoplásicos/uso terapéutico , Apoptosis , Carcinoma Hepatocelular/inmunología , Citocinas/metabolismo , Evaluación Preclínica de Medicamentos , Humanos , Células Asesinas Naturales/efectos de los fármacos , Neoplasias Hepáticas/inmunología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Ratones Endogámicos C57BL , FN-kappa B/metabolismo , Niacinamida/farmacología , Niacinamida/uso terapéutico , Compuestos de Fenilurea/uso terapéutico , SorafenibRESUMEN
The aims of this study were to evaluate the discernibility of the LIR (lateral intercondylar ridge) and the LBR (lateral bifurcate ridge) and show their reliability in femoral tunnel placement in ACL (anterior cruciate ligament) reconstruction. Additionally, their position to the femoral axis, their course, and the ACL footprint were analyzed. For this study, 235 human femora were evaluated. Of these, 166 specimens originated from the Museum of Natural History (group A), and 69 were obtained from fixed cadavers at the Anatomic Institute (group B). The femoral footprint and the osseous landmarks were identified macroscopically and labeled in the photographs. A coordinate system was outlined, and the dimensions, position, and orientation of the femoral footprint of the ACL were measured. The LBR was found in 24.7% of the specimens in group A and in only 13.2% of the specimens in group B. The LIR was found in 97.9% and 85.3% of the specimens in groups A and B, respectively. The area of the ACL footprint was 127.21 ± 32.54 mm(2) in group A and 119.58 ± 34.84 mm(2) in group B. The shapes and angles of the osseous landmarks near the line of Blumensaat were highly variable. The LBR is an unreliable intraoperative landmark for arthroscopic ACL reconstruction due to its low incidence. Other anatomical structures, such as the LIR or the osteochondral border, may be more helpful and reliable landmarks to guide proper tunnel placement.
Asunto(s)
Puntos Anatómicos de Referencia/anatomía & histología , Ligamento Cruzado Anterior/anatomía & histología , Fémur/anatomía & histología , Reconstrucción del Ligamento Cruzado Anterior , HumanosRESUMEN
Tomato (Solanum lycopersicum L.) is a widely cultivated horticultural crop. It belongs to the Solanaceae family and is known for its high concentration of essential nutrients, including vitamins, minerals, and bioactive compounds with antioxidant properties. The Mediterranean countries, including Italy, Spain, and Greece, have a diverse range of tomato landraces. Assessing the nutritional and bioactive composition of different tomato varieties and their ripening stages is crucial to determine their suitability for the market. Therefore, the aim of this study was to investigate the effect of ripening on nutritional composition (including carotenoids and polyphenols content) and antioxidant activities of fruits of three specific tomato varieties grown in Spain: Josefina and Karelya, which are cherry-like tomatoes, and Muchamiel, a type of salad tomato. In addition to evaluating their characteristics and composition (including carotenoids and polyphenol content), the antioxidant activities of these varieties at three different ripening stages were quantified. As expected, the results reveal that, as the tomatoes matured, their antioxidant capacity increased along with higher levels of carotenoids and polyphenols. Interestingly, cherry-like tomatoes showed a higher antioxidant activity than the salad tomatoes. This investigation emphasizes the role of fruit ripening in increasing carotenoid levels, which contribute to the antioxidant activity of three tomato varieties.