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Background: Even though liver kinase B1 (LKB1) is usually described as a tumor suppressor in a wide variety of tissues, it has been shown that LKB1 aberrant expression is associated with bad prognosis in Hepatocellular Carcinoma (HCC). Methods: Herein we have overexpressed LKB1 in human hepatoma cells and by using histidine pull-down assay we have investigated the role of the hypoxia-related post-translational modification of Small Ubiquitin-related Modifier (SUMO)ylation in the regulation of LKB1 oncogenic role. Molecular modelling between LKB1 and its interactors, involved in regulation of LKB1 nucleocytoplasmic shuttling and LKB1 activity, was performed. Finally, high affinity SUMO binding entities-based technology were used to validate our findings in a pre-clinical mouse model and in clinical HCC. Findings: We found that in human hepatoma cells under hypoxic stress, LKB1 overexpression increases cell viability and aggressiveness in association with changes in LKB1 cellular localization. Moreover, by using site-directed mutagenesis, we have shown that LKB1 is SUMOylated by SUMO-2 at Lys178 hampering LKB1 nucleocytoplasmic shuttling and fueling hepatoma cell growth. Molecular modelling of SUMO modified LKB1 further confirmed steric impedance between SUMOylated LKB1 and the STe20-Related ADaptor cofactor (STRADα), involved in LKB1 export from the nucleus. Finally, we provide evidence that endogenous LKB1 is modified by SUMO in pre-clinical mouse models of HCC and clinical HCC, where LKB1 SUMOylation is higher in fast growing tumors. Interpretation: Overall, SUMO-2 modification of LKB1 at Lys178 mediates LKB1 cellular localization and its oncogenic role in liver cancer. Fund: This work was supported by grants from NIH (US Department of Health and Human services)-R01AR001576-11A1 (J.M.M and M.L.M-C.), Gobierno Vasco-Departamento de Salud 2013111114 (to M.L.M.-C), ELKARTEK 2016, Departamento de Industria del Gobierno Vasco (to M.L.M.-C), MINECO: SAF2017–87301-R and SAF2014–52097-R integrado en el Plan Estatal de Investigación Cientifica y Técnica y Innovación 2013–2016 cofinanciado con Fondos FEDER (to M.L.M.-C and J.M.M., respectively), BFU2015–71017/BMC MINECO/FEDER, EU (to A.D.Q. and I.D.M.), BIOEF (Basque Foundation for Innovation and Health Research): EITB Maratoia BIO15/CA/014; Instituto de Salud Carlos III:PIE14/00031, integrado en el Plan Estatal de Investigación Cientifica y Técnica y Innovacion 2013–2016 cofinanciado con Fondos FEDER (to M.L.M.-C and J.M.M), Asociación Española contra el Cáncer (T.C.D, P·F-T and M.L.M-C), Daniel Alagille award from EASL (to T.C.D), Fundación Científica de la Asociación Española Contra el Cancer (AECC Scientific Foundation) Rare Tumor Calls 2017 (to M.L.M and M.A), La Caixa Foundation Program (to M.L.M), Programma di Ricerca Regione-Università 2007–2009 and 2011–2012, Regione Emilia-Romagna (to E.V.), Ramón Areces Foundation and the Andalusian Government (BIO-198) (A.D.Q. and I.D.M.), ayudas para apoyar grupos de investigación del sistema Universitario Vasco IT971–16 (P.A.), MINECO:SAF2015–64352-R (P.A.), Institut National du Cancer, FRANCE, INCa grant PLBIO16–251 (M.S.R.), MINECO - BFU2016–76872-R to (E.B.). Work produced with the support of a 2017 Leonardo Grant for Researchers and Cultural Creators, BBVA Foundation (M.V-R). Finally, Ciberehd_ISCIII_MINECO is funded by the Instituto de Salud Carlos III. We thank MINECO for the Severo Ochoa Excellence Accreditation to CIC bioGUNE (SEV-2016-0644). Funding sources had no involvement in study design; in the collection, analysis, and interpretation of data; in the writing of the report; and in the decision to submit the paper for publication. Keywords: LKB1, SUMO, HCC, SIRT1, STRADα

Background:Even though liver kinase B1 (LKB1) is usually described as a tumor suppressor in a wide variety of tissues, it has been shown that LKB1 aberrant expression is associated with bad prognosis in Hepatocellular Carcinoma (HCC). Methods:Herein we have overexpressed LKB1 in human hepatoma cells and by using histidine pull-down assay we have investigated the role of the hypoxia-related post-translational modification of Small Ubiquitin-related Modifier (SUMO)ylation in the regulation of LKB1 oncogenic role. Molecular modelling between LKB1 and its interactors, involved in regulation of LKB1 nucleocytoplasmic shuttling and LKB1 activity, was performed. Finally, high affinity SUMO binding entities-based technology were used to validate our findings in a pre-clinical mouse model and in clinical HCC. Findings:We found that in human hepatoma cells under hypoxic stress, LKB1 overexpression increases cell viability and aggressiveness in association with changes in LKB1 cellular localization. Moreover, by using site-directed mutagenesis, we have shown that LKB1 is SUMOylated by SUMO-2 at Lys178 hampering LKB1 nucleocytoplasmic shuttling and fueling hepatoma cell growth. Molecular modelling of SUMO modified LKB1 further confirmed steric impedance between SUMOylated LKB1 and the STe20-Related ADaptor cofactor (STRADα), involved in LKB1 export from the nucleus. Finally, we provide evidence that endogenous LKB1 is modified by SUMO in pre-clinical mouse models of HCC and clinical HCC, where LKB1 SUMOylation is higher in fast growing tumors. Interpretation:Overall, SUMO-2 modification of LKB1 at Lys178 mediates LKB1 cellular localization and its oncogenic role in liver cancer. Keywords:HCC; LKB1; SIRT1; STRADα; SUMO.

The most important sources of pollution in urban rivers are the sewage system discharges, disturbing the quality and behavior patterns of water resources. The Tunjuelo River is one of the most polluted and important stream from Bogota city, receiving numerous wastewater discharges. Therefore, its sanitation has become one of the main objectives of the environmental authority. However, in order to take action is necessary to understand the water quality behavior specifically loads patterns. This article shows the results of the load multiplier factors for total suspended solids (TSS) and biochemical oxygen demand (BOD), these factors were calculated to understand the behavior of wastewater discharges and at different points on the river. Finally, each discharge has its own characteristics that affect differently the behavior of the river downstream, resulting in a distortion in the flow pattern in most of the sampled points. Resumen Las fuentes puntuales de contaminación más importantes en los ríos urbanos son las descargas directas del sistema de alcantarillado, además estas afectan la calidad y el patrón de comportamiento de las fuentes hídricas. El río Tunjuelo es una de las corrientes de agua más importante y contaminada de la ciudad de Bogotá, pues recibe una gran cantidad de vertimientos. Por lo anterior su saneamiento se ha convertido en uno de los principales objetivos de la autoridad ambiental, sin embargo, para poder tomar acciones es necesario entender su comportamiento. En el presente artículo se muestran los resultados de los factores multiplicadores de carga para sólidos suspendidos totales (SST) y demanda bioquímica de oxígeno (DBO), puesto que estos son el instrumento económico para el cálculo de la tasa retributiva en Colombia, los cuales fueron calculados para entender el comportamiento de las descargas de aguas residuales y el de diferentes puntos sobre el río. Finalmente, se puede observar que cada descarga tiene características propias que afectan de diferente manera el comportamiento del río aguas abajo, generando como resultado una distorsión en el patrón de flujo de los factores multiplicadores en la mayoría de puntos muestreados.

Poca información existe en la literatura acerca de la calidad química referente a subproductos de desinfección (SPD) y su relación con la microbiología del agua embotellada. Por tanto, se evaluó el contenido de trihalometanos (THM) y de ácidos haloacéticos (AHA) como principales SPD en siete marcas de agua embotellada del mercado colombiano, al igual que la presencia de indicadores microbiológicos, enterobacterias, aerobios mesófilos, hongos y levaduras. Los resultados mostraron valores máximos de 135 y 140 mg/l de THM y AHA totales, así como incumplimiento del 28% de la norma propuesta por la FDA. Se encontró la presencia de alguno de los indicadores microbiológicos en el 69% de las muestras e incumplimiento de la norma colombiana de agua potable en el 30%. La relación entre la cantidad de SPD y la calidad microbiológica fue diversa, observándose un escenario recomendable de baja concentración de SPD y microorganismos en dos de las marcas evaluadas. Finalmente, se requiere mayor información para analizar el efecto de la presencia de levaduras como indicador de cambios organolépticos en el agua y su posible relación con la proliferación de otro tipo de microorganismos.

Mass spectrometry (MS) has become the method of choice for the large-scale analysis of protein ubiquitylation. There exist a number of proposed methods for mapping ubiquitin sites, each with different pros and cons. We present here a protocol for the MS analysis of the ubiquitin-proteome captured by TUBEs and subsequent data analysis. Using dedicated software and algorithms, specific information on the presence of ubiquitylated peptides can be obtained from the MS search results. In addition, a quantitative and functional analysis of the ubiquitylated proteins and their interacting partners helps to unravel the biological and molecular processes they are involved in.

Automatic analysis of increasingly growing literature repositories including data integration to other databases is a powerful tool to propose hypothesis that can be used to plan experiments to validate or disprove the hypothesis. Furthermore, it provides means to evaluate the redundancy of research line in comparison to the published literature. This is potentially beneficial for those developing research in a specific disease which are interested in exploring a particular pathway or set of genes/proteins. In the scope of the integrating book a case will be made addressing proteostasis factors in cancer. The maintenance of proteome homeostasis, known as proteostasis, is a process by which cells regulate protein translation, degradation, subcellular localization, and protein folding and consists of an integrated network of proteins. The ubiquitin-proteasome system plays a key role in essential biological processes such as cell cycle, DNA damage repair, membrane trafficking, and maintaining protein homeostasis. Cells maintain proteostasis by regulating protein translation, degradation, subcellular localization, and protein folding. Aberrant proteostasis leads to loss-of-function diseases (cystic fibrosis) and gain-of-toxic-function diseases (Alzheimer's, Parkinson's, and Huntington's disease). Cancer therapy on the other hand explores inhibition of proteostasis factors to trigger endoplasmic reticulum stress with subsequent apoptosis. Alternatively therapies target deubiquitinases and thereby regulate tumor promoters or suppressors. Furthermore, mutations in specific proteostasis factors are associated with higher risk for specific cancers, e.g., BRCA mutations in breast cancer. This chapter discusses proteostasis protein factors' association with cancer from a literature mining perspective.

This paper presents an exploratory study of pulp mill bleaching effluent treatment by a biological–photocatalytic coupled system. A fungus, Trametes pubescens, immobilized on polyurethane foam was used to inoculate the biological pre-treatment system. The pretreated effluent was then exposed to a photocatalytic treatment in which two catalysts (TiO2 and ZnO) and two supports (aluminum foil and Luffa cylindrica) were tested. Catalyst characterization was carried out by means of X-ray Diffraction (XRD) and Scanning Electron Microscopy (SEM). Information about crystalline structure, chemical composition, morphology, homogeneity and distribution on the support surface area was obtained. The overall biological–photocatalytic coupled system achieved degradation of 96% of initial total organic carbon (TOC), 97% of 2-chlorophenol (2-CP), 90% of 2,4-dichlorophenol (2,4-CP) and 99% of 2,4,6-trichlorophenol (2,4,6-TCP). This approach of synergistic coupling of T. pubescens and a semiconductor photocatalyst appears to be a viable alternative for the treatment of these non-biodegradable effluents.

The aim of this study was to evaluate the effect of the macrophyte waterhyacinth on the water quality of a reservoir in Colombia. Postinvasion water quality assessments were performed during three different hydrological periods and compared with preinvasion data. The results show that water quality has improved, especially in terms of transparency, showing an increase in Secchi disk values from 0.3 to 2 m (1 to 6.6 ft). The analysis strongly suggests that the improvement in water quality is due to the filtering and precipitation effects provided by waterhyacinth. This case highlights the dilemma of managing an invasive species that provides a beneficial ecological service while posing a high ecological risk. Nomenclature: Waterhyacinth, Eichhornia crassipes (Mart.) Solms.

Numerical and computational modelling of flow and pollutant dynamics in urban drainage systems is becoming more and more integral to planning and design. The main aim of integrated flow and pollutant models is to quantify the efficiency of different measures at reducing the amount of pollutants discharged into receiving water bodies and minimise the consequent negative water quality impact. The open source toolbox CITY DRAIN developed in the Matlab/Simulink© environment, which was designed for integrated modelling of urban drainage systems, is used in this work. The goal in this study was to implement and test computational routines for representing sediment and pollutant loads in order to evaluate catchment surface pollution. Tested models estimate the accumulation, erosion and transport of pollutants—aggregately—on urban surfaces and in sewers. The toolbox now includes mathematical formulations for accumulation of pollutants during dry weather period and their wash-off during rainfall events. The experimental data acquired in a previous research project carried out by the Environmental Engineering Research Centre (CIIA) at the Universidad de los Andes in Bogotá (Colombia) was used for the calibration of the models. Different numerical approaches were tested for their ability to calibrate to the sediment transport conditions. Initial results indicate, when there is more than one peak during the rainfall event duration, wash-off processes probably can be better represented using a model based on the flow instead of the rainfall intensity. Additionally, it was observed that using more detailed models (compared with an instantaneous approach) for representing pollutant accumulation do not necessarily lead to better results.

In developing countries, lack of sanitation coverage and continuously growing populations are increasing the pressures on receiving waters. In the context of Bogotá (Colombia), this paper presents an overview of earlier, recent and ongoing research towards improved management of urban drainage systems using an integrated modelling framework. Research results have shown that there is a need to assess the urban drainage system as one entity, when considering pollution control objectives, and optimum management and operation. This holistic approach offers an opportunity to investigate the interactions among sub-systems and the impact of the whole system on the river water quality. En países en vía de desarrollo, la falta de infraestructura de saneamiento básico y el número creciente de habitantes en sus centros urbanos han venido aumentando las demandas sobre los recursos hídricos disponibles. En el contexto de Bogotá (Colombia), este artículo emplea un marco de modelación integrada y presenta una revisión de estudios que han buscado una mejora en el manejo de los sistemas de drenaje urbano. Es sabido que hay una necesidad de evaluar estos sistemas como una única entidad cuando se busca propender por el control de la contaminación, su manejo y operación óptima. Esta visión holística ofrece la oportunidad de estudiar las interacciones entre los sub-sistemas y el impacto de la totalidad del sistema en la calidad del agua de los cuerpos receptores.

In developing countries, lack of sanitation coverage and continuously growing populations are increasing the pressures on receiving waters. In the context of Bogotá (Colombia), this paper presents an overview of earlier, recent and ongoing research towards improved management of urban drainage systems using an integrated modelling framework. Research results have shown that there is a need to assess the urban drainage system as one entity, when considering pollution control objectives, and optimum management and operation. This holistic approach offers an opportunity to investigate the interactions among sub-systems and the impact of the whole system on the river water quality. / En países en vía de desarrollo, la falta de infraestructura de saneamiento básico y el número creciente de habitantes en sus centros urbanos han venido aumentando las demandas sobre los recursos hídricos disponibles. En el contexto de Bogotá (Colombia), este artículo emplea un marco de modelación integrada y presenta una revisión de estudios que han buscado una mejora en el manejo de los sistemas de drenaje urbano. Es sabido que hay una necesidad de evaluar estos sistemas como una única entidad cuando se busca propender por el control de la contaminación, su manejo y operación óptima. Esta visión holística ofrece la oportunidad de estudiar las interacciones entre los sub-sistemas y el impacto de la totalidad del sistema en la calidad del agua de los cuerpos receptores.

Un sistema de microfiltración a escala laboratorio fue estudiado como alternativa de potabilización para el agua de tres diferentes embalses de la Sabana de Bogotá, Colombia. El desempeño del sistema fue evaluado basándose en la eficiencia de eliminación de parámetros clásicos, en el comportamiento del flujo de permeado y en el grado de recuperación del flujo después de la limpieza de la membrana. Ensayos adicionales fueron realizados adicionando CAP al agua de alimentación. Valores de presión de trabajo más bajos resultaron en velocidades de ensuciamiento menores y la adición de CAP redujo la tendencia al ensuciamiento irreversible de la membrana. El sistema de microfiltración directa es una alternativa potencial para la potabilización de agua superficial, pero estudios más profundos son indispensables para corroborar a largo término la idoneidad del proceso. / A microfiltration lab-scale system was studied as alternative to water purification of three reservoirs from the Sabana of Bogotá, Colombia. The system performance was evaluated based on the efficiency of classic parameters elimination, the flux behavior and the degree of flux recovery after membrane cleaning. Additional tests were made adding CAP to the feeding water. Lower work pressures lead to lower fouling membrane rates and the CAP addition reduced the irreversible fouling. Direct microfiltration system is a potential alternative for superficial water purification, but deeper studies are required to corroborate the process suitability to long term.

By controlling HIFα hydroxylation and stability, the prolyl hydroxylase domain (PHD)-containing proteins are essential to the maintenance of oxygen homeostasis; therefore these enzymes are tightly regulated. Small ubiquitin-like modifier (SUMO) is a 10-kDa protein readily conjugated to lysine residues of the targeted proteins in a process termed SUMOylation. In this study, we introduce SUMO conjugation as a novel regulator of PHD3 (also known as EGLN3). PHD3 SUMOylation occurs at a cluster of four lysines at the C-terminal end of the protein. Furthermore, PHD3 SUMOylation by SUMO2 or SUMO3 contributes to PHD3-mediated repression of HIF1-dependent transcriptional activity. Interestingly, PHD3-SUMO conjugation does not affect PHD3 hydroxylase activity or HIF1α stability, providing new evidence for a dual role of PHD3 in HIF1 regulation. Moreover, we show that hypoxia modulates PHD3-SUMO conjugation and that this modification inversely correlates with HIF1 activation. PHD3 SUMOylation highlights a new and additional layer of regulation that is likely required to fine-tune HIF function.

Studying postubiquitylation events has always been a difficult task due to the labile nature of these posttranslational modifications. When utilized in tandem, ubiquitin-binding entities (TUBEs) not only increase up to thousand times the affinity for poly-ubiquitin chains but also protect ubiquitylated proteins from the action of the proteasome and de-ubiquitylating enzymes.