Role of C3G in the differentiation and maturation of megakaryocytes

[ES] C3G es un activador de varios miembros de la superfamilia Ras, principalmente de Ras1 y R-Ras. Estudios anteriores sugieren que C3G juega un papel en diferentes procesos de diferenciación celular, como la diferenciación de células de neuroblastoma a neuronas o la diferenciación adipocítica, actuando a través de su principal diana Rap1. La implicación de C3G en la diferenciación y maduración megacariocítica se ha estudiado mediante el uso de dos modelos experimentales: líneas celulares hematopoyéticas K562 y HEL transfectadas con distintas construcciones de C3G que lo sobreexpresan o silencian y ratones transgénicos para C3G y C3GΔCat (mutante de C3G delecionado en el dominio catalítico o GEF), donde ambos transgenes se encuentran expresados bajo el promotor del gen PF4 (platelet factor 4), exclusivo de megacariocitos y plaquetas. Los estudios en líneas celulares, utilizando PMA como estímulo de diferenciación, han revelado que la proteína C3G tiene un papel regulador positivo en la expresión de marcadores megacariocíticos CD41 y CD61, en el incremento en la poliploidia celular mediado por p21 y en la adquisición de determinadas características morfológicas, todos ellos cambios que contribuyen al compromiso hacia el linaje megacariocítico. Dichos cambios se producen a través de las vías de señalización de Rap1 y PI3K y son independientes de la activación a través de PKC. Además, PMA, promueve la fosforilación de C3G en la Tirosina-504, lo cual se encuentra relacionado con la activación sostenida de la vía de señalización Rap1-ERK1/2 que ocurre en etapas tardías de la maduración megacariocítica. Los estudios en los modelos transgénicos para C3G y C3GΔCat, han confirmado lo observado en las líneas celulares. Células procedentes de la médula ósea de los ratones transgénicos para C3G, pero no procedentes de los ratones C3GΔCat, muestran un incremento en la expresión de CD41 y CD61 tras el tratamiento con trombopoyetina. Además, la expresión transgénica de C3G incrementa el porcentaje de megacariocitos maduros con alto contenido en ADN. Adicionalmente, mediante el uso de explantos de la médula ósea se observó un incremento en la migración de los megacariocitos maduros desde el nicho osteoblástico al vascular y una mejora en la producción de proplaquetas en los ratones tgC3G frente a sus correspondientes controles. Por el contrario los megacariocitos procedentes de explantos de médula ósea de ratones C3GCat presentaron una menor migración, así como una menor formación de proplaquetas. Estos resultados sugieren una participación de C3G en diferentes aspectos clave de la megacariopoyesis a través de un mecanismo mediado por su actividad GEF

Etiopathogenesis and pathophysiology of cholestasis

Normal hepatobiliary function depends on an adequate bile flow from the liver through the biliary tree to the gallbladder, where bile is stored and concentrated, and from the gallbladder to the duodenum when it is required for the digestive process. Interruption of this secretory function results in partial or complete cholestasis, which is accompanied by important repercussions due to the lack of bile acids in the intestine and their regurgitation from hepatocytes to blood together with potentially toxic compounds that are normally eliminated in bile. The presence of active and selective transporter proteins located at both poles of the plasma membrane of hepatocytes, cholangiocytes, and epithelial cells of the ileal mucosa, together with the ability of hepatocytes to synthesize bile acids from cholesterol, enables the so-called bile acid enterohepatic circulation, which is essential in liver and gastrointestinal tract physiology. The presence in the ducts of the biliary tree of agents reducing their luminal diameter by external compression or space-occupying obstacles, either in the duct wall or its lumen, can result in total or partial obstructive cholestasis. The clinical impact and management of cholestasis are different depending on the intrahepatic or extrahepatic location of the obstacle. Thus, surgical interventions can often be helpful in removing extrahepatic obstructions and restoring normal bile flow to the duodenum. In contrast, hepatocyte or cholangiocyte damage, either global, restricted to subcellular compartments, or more specifically affecting the elements of the canalicular secretory machinery, may result in hepatocellular cholestasis or cholangiopathies. In these cases, bile flow interruption is usually partial and, except for extremely severe cases when liver transplantation is required, these patients often treated with pharmacological agents, such as ursodeoxycholic acid (UDCA) and rifampicin. The present review gathers updated information on the etiopathogenesis and pathophysiological aspects of different types of cholestasis

C3G, through its GEF activity, induces megakaryocytic differentiation and proplatelet formation

[Background]: Megakaryopoiesis allows platelet formation, which is necessary for coagulation, also playing an important role in different pathologies. However, this process remains to be fully characterized. C3G, an activator of Rap1 GTPases, is involved in platelet activation and regulates several differentiation processes. [Methods]: We evaluated C3G function in megakaryopoiesis using transgenic mouse models where C3G and C3GΔCat (mutant lacking the GEF domain) transgenes are expressed exclusively in megakaryocytes and platelets. In addition, we used different clones of K562, HEL and DAMI cell lines with overexpression or silencing of C3G or GATA-1. [Results]: We found that C3G participates in the differentiation of immature hematopoietic cells to megakaryocytes. Accordingly, bone marrow cells from transgenic C3G, but not those from transgenic C3GΔCat mice, showed increased expression of the differentiation markers CD41 and CD61, upon thrombopoietin treatment. Furthermore, C3G overexpression increased the number of CD41+ megakaryocytes with high DNA content. These results are supported by data obtained in the different models of megakaryocytic cell lines. In addition, it was uncovered GATA-1 as a positive regulator of C3G expression. Moreover, C3G transgenic megakaryocytes from fresh bone marrow explants showed increased migration from the osteoblastic to the vascular niche and an enhanced ability to form proplatelets. Although the transgenic expression of C3G in platelets did not alter basal platelet counts, it did increase slightly those induced by TPO injection in vivo. Moreover, platelet C3G induced adipogenesis in the bone marrow under pathological conditions. [Conclusions]: All these data indicate that C3G plays a significant role in different steps of megakaryopoiesis, acting through a mechanism dependent on its GEF activity.This work was supported by grants from the Spanish Ministry of Economy and Competitiveness [SAF2013–48210-C2–1-R and SAF2016–76588-C2–2-R to CG, SAF2013–48210-C2–2-R and SAF2016–76588-C2–1-R to AP], and by two grants from the Council of Education of Junta de Castilla y León, Spain [SA157A12–1 and SA017U16 to CG]. All funding was cosponsored by the European FEDER Program

LCA of greywater management within a water circular economy restorative thinking framework

Greywater reuse is an attractive option for the sustainable management of water under water scarcity circumstances, within a water circular economy restorative thinking framework. Its successful deployment relies on the availability of low cost and environmentally friendly technologies. The life cycle assessment (LCA) approach provides the appropriate methodological tool for the evaluation of alternative treatments based on environmental decision criteria and, therefore, it is highly useful during the process conceptual design. This methodology should be employed in the early design phase to select those technologies with lower environmental impact. This work reports the comparative LCA of three scenarios for greywater reuse: photocatalysis, photovoltaic solar-driven photocatalysis and membrane biological reactor, in order to help the selection of the most environmentally friendly technology. The study has been focused on the removal of the surfactant sodium dodecylbenzenesulfonate, which is used in the formulation of detergents and personal care products and, thus, widely present in greywater. LCA was applied using the Environmental Sustainability Assessment methodology to obtain two main environmental indicators in order to simplify the decision making process: natural resources and environmental burdens. Energy consumption is the main contributor to both indicators owing to the high energy consumption of the light source for the photocatalytic greywater treatment. In order to reduce its environmental burdens, the most desirable scenario would be the use of solar light for the photocatalytic transformation. However, while the technological challenge of direct use of solar light is approached, the environmental suitability of the photovoltaic solar energy driven photocatalysis technology to greywater reuse has been demonstrated, as it involves the smallest environmental impact among the three studied alternatives.Financial support from the Spanish Ministry of Economy and Competitiveness and from FEDER funds for projects CTM2013-43539-R, CTM2015-69845-R and CTQ2015-66078-R(MINECO/FEDER, UE) is gratefully acknowledged. Sara Dominguez and Jara Laso would also like to express their gratitude for the FPI postgraduate research grants (BES-2013-064055 and BES-2014-069368)

C3G promotes a selective release of angiogenic factors from activated mouse platelets to regulate angiogenesis and tumor metastasis

[EN]Previous observations indicated that C3G (RAPGEF1) promotes α-granule release, evidenced by the increase in P-selectin exposure on the platelet surface following its activation. The goal of the present study is to further characterize the potential function of C3G as a modulator of the platelet releasate and its implication in the regulation of angiogenesis. Proteomic analysis revealed a decreased secretion of anti-angiogenic factors from activated transgenic C3G and C3GΔCat platelets. Accordingly, the secretome from both transgenic platelets had an overall pro-angiogenic effect as evidenced by an in vitro capillary-tube formation assay with HUVECs (human umbilical vein endothelial cells) and by two in vivo models of heterotopic tumor growth. In addition, transgenic C3G expression in platelets greatly increased mouse melanoma cells metastasis. Moreover, immunofluorescence microscopy showed that the pro-angiogenic factors VEGF and bFGF were partially retained into α-granules in thrombin- and ADP-activated mouse platelets from both, C3G and C3GΔCat transgenic mice. The observed interaction between C3G and Vesicle-associated membrane protein (Vamp)-7 could explain these results. Concomitantly, increased platelet spreading in both transgenic platelets upon thrombin activation supports this novel function of C3G in α-granule exocytosis. Collectively, our data point out to the co-existence of Rap1GEF-dependent and independent mechanisms mediating C3G effects on platelet secretion, which regulates pathological angiogenesis in tumors and other contexts. The results herein support an important role for platelet C3G in angiogenesis and metastasis

Influence of radiation and TiO2 concentration on the hydroxyl radicals generation in a photocatalytic LED reactor. Application to dodecylbenzenesulfonate degradation

One of the main issues associated to the development of photocatalysis is the lack of adequate indexes that allow the comparison of the results obtained in different experimental setup designs. The hydroxyl radicals (o OH) generation rate is a key factor to determine the overall oxidation rate. In this work, using a Light Emitting Diodes (LEDs) reactor aimed to maximize light ef?ciency and minimize energy consumption, the o OH generated have been determined as a function of the radiation and catalyst concentration following an indirect method based on the reaction between o OH and dimethyl sulfoxide (DMSO) to produce formaldehyde. Finally, the methodology has been applied to analyze the degradation kinetics of the anionic surfactant dodecylbenzenesulfonate (DBS), frequently used in shampoo formulations and detergents for washing machines. We propose a method based on the indirect determination of o OH radicals generation rate that allows the assessment and comparison of the kinetics of photocatalytic oxidation of pollutants.Financial support from the Spanish Ministry of Economy and Competitiveness and from FEDER funds (projects CTM2012-33917 and CTQ2012-31639) are gratefully acknowledged

Aportaciones a los estudios económico-administrativos. Reflexiones teóricas y evidencias empíricas

La economía y la administración están inmersas en el universo de las ciencias sociales, áreas del conocimiento que comprenden dominios diferentes, pero también convergen y se intersectan en un objeto de estudio: las organizacienones y sus agregados (industrias o sectores). Desde una perspectiva social, los estudios económico-administrativos informan sobre cómo hacer eficientes a las organizaciones y competitivos a los sectores con la finalidad de impactar positivamente en la justicia social (por ejemplo: mediante productos de mejor calidad a mejores precios). El auge de este tipo de estudios está extendido a fenómenos clave en el mundo de hoy, tales como: la ventaja competitiva, la innovación, la administración de la cadena de suministros y la administración verde, entre otros.El objetivo de este libro es difundir las aportaciones hechas a los estudios económico-administrativos, ya sea como reflexiones teóricas o a modo de evidencia empírica, pues la conjunción de varias ciencias ayuda a comprender fenómenos complejos mejor que una disciplina en forma aislada. Más que la homogeneidad de teorías o hegemonía de unas sobre otras, la investigación interdisciplinaria construye un acercamiento a la diversidad de puntos de vista: se buscan factores, métodos o lenguajes comunes alrededor de un objeto de estudio (Fernández, 2009; Martínez, 1997), llegándose a conformar una comunidad científica con antecedentes disciplinarios diferentes, pero con cierta unidad, relaciones y acciones recíprocas (Kuhn, 1995). Así, en este texto el objeto de estudio es el funcionamiento e interacción de las organizaciones, como integrantes de los sectores económicos, en los mercados locales y globales. La comunidad científica está conformada por investigadores y egresados del Doctorado en Ciencias Económico-Administrativas, el cual nace en 2006 en la Universidad Autónoma del Estado de México, a cargo de la Dependencia de Educación Superior (DES) Económico-Administrativa

Cabbage and fermented vegetables : From death rate heterogeneity in countries to candidates for mitigation strategies of severe COVID-19

Large differences in COVID-19 death rates exist between countries and between regions of the same country. Some very low death rate countries such as Eastern Asia, Central Europe, or the Balkans have a common feature of eating large quantities of fermented foods. Although biases exist when examining ecological studies, fermented vegetables or cabbage have been associated with low death rates in European countries. SARS-CoV-2 binds to its receptor, the angiotensin-converting enzyme 2 (ACE2). As a result of SARS-CoV-2 binding, ACE2 downregulation enhances the angiotensin II receptor type 1 (AT(1)R) axis associated with oxidative stress. This leads to insulin resistance as well as lung and endothelial damage, two severe outcomes of COVID-19. The nuclear factor (erythroid-derived 2)-like 2 (Nrf2) is the most potent antioxidant in humans and can block in particular the AT(1)R axis. Cabbage contains precursors of sulforaphane, the most active natural activator of Nrf2. Fermented vegetables contain many lactobacilli, which are also potent Nrf2 activators. Three examples are: kimchi in Korea, westernized foods, and the slum paradox. It is proposed that fermented cabbage is a proof-of-concept of dietary manipulations that may enhance Nrf2-associated antioxidant effects, helpful in mitigating COVID-19 severity.Peer reviewe

Nrf2-interacting nutrients and COVID-19 : time for research to develop adaptation strategies

There are large between- and within-country variations in COVID-19 death rates. Some very low death rate settings such as Eastern Asia, Central Europe, the Balkans and Africa have a common feature of eating large quantities of fermented foods whose intake is associated with the activation of the Nrf2 (Nuclear factor (erythroid-derived 2)-like 2) anti-oxidant transcription factor. There are many Nrf2-interacting nutrients (berberine, curcumin, epigallocatechin gallate, genistein, quercetin, resveratrol, sulforaphane) that all act similarly to reduce insulin resistance, endothelial damage, lung injury and cytokine storm. They also act on the same mechanisms (mTOR: Mammalian target of rapamycin, PPAR gamma:Peroxisome proliferator-activated receptor, NF kappa B: Nuclear factor kappa B, ERK: Extracellular signal-regulated kinases and eIF2 alpha:Elongation initiation factor 2 alpha). They may as a result be important in mitigating the severity of COVID-19, acting through the endoplasmic reticulum stress or ACE-Angiotensin-II-AT(1)R axis (AT(1)R) pathway. Many Nrf2-interacting nutrients are also interacting with TRPA1 and/or TRPV1. Interestingly, geographical areas with very low COVID-19 mortality are those with the lowest prevalence of obesity (Sub-Saharan Africa and Asia). It is tempting to propose that Nrf2-interacting foods and nutrients can re-balance insulin resistance and have a significant effect on COVID-19 severity. It is therefore possible that the intake of these foods may restore an optimal natural balance for the Nrf2 pathway and may be of interest in the mitigation of COVID-19 severity

Omecamtiv mecarbil in chronic heart failure with reduced ejection fraction, GALACTIC‐HF: baseline characteristics and comparison with contemporary clinical trials

Aims: The safety and efficacy of the novel selective cardiac myosin activator, omecamtiv mecarbil, in patients with heart failure with reduced ejection fraction (HFrEF) is tested in the Global Approach to Lowering Adverse Cardiac outcomes Through Improving Contractility in Heart Failure (GALACTIC‐HF) trial. Here we describe the baseline characteristics of participants in GALACTIC‐HF and how these compare with other contemporary trials. Methods and Results: Adults with established HFrEF, New York Heart Association functional class (NYHA) ≥ II, EF ≤35%, elevated natriuretic peptides and either current hospitalization for HF or history of hospitalization/ emergency department visit for HF within a year were randomized to either placebo or omecamtiv mecarbil (pharmacokinetic‐guided dosing: 25, 37.5 or 50 mg bid). 8256 patients [male (79%), non‐white (22%), mean age 65 years] were enrolled with a mean EF 27%, ischemic etiology in 54%, NYHA II 53% and III/IV 47%, and median NT‐proBNP 1971 pg/mL. HF therapies at baseline were among the most effectively employed in contemporary HF trials. GALACTIC‐HF randomized patients representative of recent HF registries and trials with substantial numbers of patients also having characteristics understudied in previous trials including more from North America (n = 1386), enrolled as inpatients (n = 2084), systolic blood pressure < 100 mmHg (n = 1127), estimated glomerular filtration rate < 30 mL/min/1.73 m2 (n = 528), and treated with sacubitril‐valsartan at baseline (n = 1594). Conclusions: GALACTIC‐HF enrolled a well‐treated, high‐risk population from both inpatient and outpatient settings, which will provide a definitive evaluation of the efficacy and safety of this novel therapy, as well as informing its potential future implementation