Filtration assay in Corbicula fluminea (Müller, 1774) for assessing the toxicity of PCPs

[Resumo]: Os produtos de coidado persoal (PCPs) son contaminantes emerxentes presentes de forma habitual nos medios acuáticos. Teñen efectos negativos para o ecosistema e a saúde humana actuando principalmente como disruptores endócrinos. Ademais adoitan ser bioacumulados en organismos como Corbicula fluminea, un bivalvo bentónico invasor e filtrador empregado frecuentemente como bioindicador de contaminación mediante biomarcadores moleculares. Neste traballo emprégase a inhibición da filtración como biomarcador, para testar os efectos tóxicos dos PCPs, cuantificándoa a través da porcentaxe de remoción de algas e a taxa de aclaramento per cápita (CRmáx). A porcentaxe de remoción das algas foi do 100% nos primeiros 30 minutos, o que condicionou a CRmáx dos individuos. Aparentemente o TCS e a BP-4 non teñen efectos na taxa de filtración de C. fluminea, resultado que podería estar causado polo deseño do método experimental.[Resumen]: Los productos de cuidado personal (PCPs) son contaminantes emergentes presentes de forma habitual en los medios acuáticos. Tienen efectos negativos para el ecosistema y la salud humana actuando principalmente como disruptores endócrinos. Además suelen ser bioacumulados en organismos como Corbicula fluminea, un bivalvo bentónico invasor y filtrador empleado frecuentemente como bioindicador de contaminación mediante biomarcadores moleculares. En este trabajo se ha usado la inhibición da filtración como biomarcador, para testar los efectos tóxicos de los PCPs, cuantificándola a través del porcentaje de remoción de algas y la tasa de aclarado per cápita (CRmáx). El porcentaje de remoción das algas fue del 100% en los primeiros 30 minutos, lo que condicionó la CRmáx de los individuos. Aparentemente el TCS y la BP-4 no tienen efectos en la tasa de filtración de C. fluminea, resultado que podría estar causado por el diseño del método experimental.[Abstract]: Personal care products (PCPs) are emerging contaminants commonly found in aquatic environments. They have negative effects for the ecosystem and human health acting mainly as endocrine disruptors. Moreover, they are usually bioaccumulated in organisms such as Corbicula fluminea, an invasive benthic bivalve and filter feeder 2 frequently used as a bioindicator of contamination by molecular biomarkers. In this work, the inhibition of filtration as a biomarker has been used to test the toxic effects of PCPs, quantifying it through the percentage of algae removal and the clearence rate per capita (CRmax). The percentage of removal of algae was 100% in the first 30 minutes, which conditioned the CRmax of the individuals. Apparently, the TCS and the BP-4 have no effects on the filtration rate of C. fluminea, a result that could be caused by the design of the experimental method.Traballo fin de grao (UDC.CIE). Bioloxía. Curso 2018/201

Modeling the Breeding Habitat of Charadrius Alexandrinus (Linnaeus, 1758) in Galician Coast From Satellite Images

[Resumo] Os rápidos cambios ambientais que se están a producir a nivel global afectan de forma directa á distribución e abundancia das especies. Isto converte á conservación nun dos principais obxectivos da ecoloxía moderna. Detectar a tempo estes cambios é fundamental á hora de establecer medidas efectivas de conservación. A teledetección representa unha ferramenta imprescindible, pero aínda pouco desenvolta, para o estudo a diferentes escalas espaciais e temporais, do hábitat reprodutor de especies ameazadas. Neste traballo modelizamos o hábitat reprodutor da píllara das dunas (Charadrius alexandrinus), a cal presenta tendencias poboacionais que indican un forte declive a curto prazo en España. A partir do arquivo histórico da misión Landsat, obtivéronse índices espectrais de vexetación e contido en auga que permitiron caracterizar anualmente cada praia de posible nidificación. Estas variables foron incluídas como preditores nun modelo lineal xeneralizado mixto, co obxectivo de explicar a variación especiotemporal no número de parellas da píllara das dunas en Galicia nun período de 22 anos (1999-2020). Os resultados confirman que as variables preditoras obtidas por teledetección, que caracterizan a vexetación, o contido en auga ou a reflectividade do solo, presentan unha alta capacidade explicativa da variación no número de parellas nidificantes de píllara das dunas na costa galega (R2 marxinal = 0,5; R2 condicional = 0,757). Das 11 variables non correlacionadas incluídas no modelo, 7 resultaron significativas, 4 fan referencia ao contido en auga (MSAVI_mean, NDWI_min, MNDWI_range e NDWI2_sd) e as 3 restantes están relacionadas co estado da vexetación e a diferenciación entre esta e o solo nu (MSAVI_mean, EVI_max e EVI2_max). Este traballo confirma a utilidade e o potencial da teledetección para modelizar as variacións espaciotemporais en parámetros chave da nidificación da píllara das dunas na costa de Galicia.[Resumen] Los rápidos cambios ambientales que se están produciendo a nivel global afectan de forma directa a la distribución y la abundancia de las especies. Esto convierte a la conservación en uno de los principales objetivos de la ecología moderna. Detectar a tiempo estos cambios es fundamental a la hora de establecer medidas eficaces para la conservación. La teledetección representa una herramienta imprescindible pero todavía poco desarrollada, para el estudio a diferentes escalas espaciales y temporales, del hábitat reproductor de especies ameazadas. En este trabajo modelizamos el hábitat reproductor del chorlitejo patinegro (Charadrius alexandrinus), el cual presenta tendencias poblacionales que indican un fuerte declive a corto plazo en España. A partir del archivo histórico de Landsat, se obtuvieron índices espectrales de vegetación y contenido en agua que permitieron caracterizar anualmente cada playa de posible nidificación. Estas variables fueron incluidas como predictores en un modelo lineal generalizado mixto, con el objetivo de explicar la variación espaciotemporal en el número de parejas de chorlitejo patinegro en Galicia en un período de 22 años (1999-2020). Los resultados confirman que las variables predictoras obtenidas por teledetección, que caracterizan la vegetación, el contenido en agua o la reflectividad del suelo, presentan una alta capacidad explicativa de la variación en el número de parejas de chorlitejo patinegro en la costa gallega (R2 marginal = 0,5; R2 condicional = 0,757). De las 11 variables no correlacionadas incluidas en el modelo, 7 resultaron significativas, 4 hacen referencia al contenido en agua (MSAVI_mean, NDWI_min, MNDWI_range e NDWI2_sd) y las 3 restantes están relacionadas con el estado de la vegetación y la diferenciación entre esta y el suelo desnudo (MSAVI_mean, EVI_max e EVI2_max). Este trabajo confirma la utilidad y el potencial de la teledetección para modelizar las variaciones espaciotemporales en parámetros clave de la nidificación del chorlitejo patinegro en la costa de Galicia.[Abstract] The rapid environmental changes taking place globally directly affect the distribution and abundance of species. This makes conservation one of the main objectives of modern ecology. Detecting these changes on time is essential when establishing effective measures for conservation. Remote sensing represents an essential but still underdeveloped tool for the study at different spatial and temporal scales of the reproductive habitat of threatened species. In this study we model the breeding habitat of the Kentish Plover (Charadrius alexandrinus), which shows population trends that indicate a strong short-term decline in Spain. From Landsat historical archive, spectral indices of vegetation and water content were obtained that allowed each beach of posible nesting to be characterized annually. These variables were included as predictors in a generalized linear mixed model, with the aim of explaining spatio-temporal variation in the number of pairs of Kentish Plover in Galicia over a 22 years period (1999-2020). The results confirm that the predictor variables obtained by remote sensing, which caracterize the vegetation, the water content or the soil reflectance, have a hight explanatory capacity of the variation in the number of pairs of Kentish Plover on the Galician coast (marginal R2 = 0.5; conditional R2 = 0.757). Of the 11 uncorrelated variables included in the model, 7 resulted singnificant, 4 of them refer to the water content (MSAVI_mean, NDWI_min, MNDWI_range e NDWI2_sd) and the remaining 3 are related to the state of vegetation and the differentiation between it and the bare soil (MSAVI_mean, EVI_max e EVI2_max). This study confirms the usefulness and potential of remote sensing to model spatio-temporal variation in key parameters of Kentish Plover nesting on the Galcian coast.Traballo fin de mestrado (UDC.CIE). Biodiversidade terrestre. Curso 2021/202

Riboflavin kinase and pyridoxine 5'-phosphate oxidase complex formation envisages transient interactions for FMN cofactor delivery

Enzymes catalysing sequential reactions have developed different mechanisms to control the transport and flux of reactants and intermediates along metabolic pathways, which usually involve direct transfer of metabolites from an enzyme to the next one in a cascade reaction. Despite the fact that metabolite or substrate channelling has been widely studied for reactant molecules, such information is seldom available for cofactors in general, and for flavins in particular. Flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) act as cofactors in flavoproteins and flavoenzymes involved in a wide range of physiologically relevant processes in all type of organisms. Homo sapiens riboflavin kinase (RFK) catalyses the biosynthesis of the flavin mononucleotide cofactor, and might directly interplay with its flavin client apo-proteins prior to the cofactor transfer. Non-etheless, none of such complexes has been characterized at molecular or atomic level so far. Here, we particularly evaluate the interaction of riboflavin kinase with one of its potential FMN clients, pyridoxine-5′-phosphate oxidase (PNPOx). The interaction capacity of both proteins is assessed by using isothermal titration calorimetry, a methodology that allows to determine dissociation constants for interaction in the micromolar range (in agreement with the expected transient nature of the interaction). Moreover, we show that; i) both proteins become thermally stabilized upon mutual interaction, ii) the tightly bound FMN product can be transferred from RFK to the apo-form of PNPOx producing an efficient enzyme, and iii) the presence of the apo-form of PNPOx slightly enhances RFK catalytic efficiency. Finally, we also show a computational study to predict likely RFK-PNPOx binding modes that can envisage coupling between the FMN binding cavities of both proteins for the potential transfer of FMN

Caracterización funcional de AIFM3 (Apoptosis-inducing factor mitochondrion-associated 3) mediante inactivación del gen por la técnica CRISPR-Cas9 y sobreexpresión de la proteína en bacterias y células eucariotas

El factor inductor de apoptosis homólogo a AIF (AIFL) es un miembro de la familia de proteínas AIF (factor inductor de apoptosis) del que se posee escasa información tanto a nivel estructural como funcional. Con el objeto de estudiar la función de esta proteína se generó una línea celular humana KO para el gen que la codifica, aifm3, por medio de la técnica de edición genómica CRIPSR/Cas9. La expresión ubicua del factor sugiere una posible función esencial en la célula, que se vio sustentada por la observación de un crecimiento significativamente ralentizado en la línea celular silenciada. En paralelo, se estudió la hipotética función pro-apoptótica del factor, diseñando vectores de expresión tanto para células procariotas como eucariotas con el fin de sobreexpresar el gen aifm3 en Escherichia coli y en la línea celular humana HEK293T. La sobreexpresión en bacterias se demostró mediante Western Blot, mientras que no se observó expresión ni efectos significantes en las células humanas transfectadas. La optimización de la expresión en bacterias permitirá, en estudios posteriores, diseñar un protocolo de purificación para su caracterización tanto bioquímica (mediante ensayos cinéticos) como estructural (por técnicas de cristalización)

Beyond a platform protein for the degradosome assembly: the apoptosis-inducing factor as an efficient nuclease involved in chromatinolysis

The Apoptosis-Inducing Factor (AIF) is a moonlighting flavoenzyme involved in the assembly of mitochondrial respiratory complexes in healthy cells, but also able to trigger DNA cleavage and parthanatos. Upon apoptotic-stimuli, AIF redistributes from the mitochondria to the nucleus, where upon association with other proteins such as endonuclease CypA and histone H2AX, it is proposed to organize a DNA–degradosome complex. In this work, we provide evidence for the molecular assembly of this complex as well as for the cooperative effects among its protein components to degrade genomic DNA into large fragments. We have also uncovered that AIF has nuclease activity that is stimulated in the presence of either Mg2+ or Ca2+. Such activity allows AIF by itself and in cooperation with CypA to efficiently degrade genomic DNA. Finally, we have identified TopIB and DEK motifs in AIF as responsible for its nuclease activity. These new findings point, for the first time, to AIF as a nuclease able to digest nuclear dsDNA in dying cells, improving our understanding of its role in promoting apoptosis and opening paths for the development of new therapeutic strategies

SETBP1 overexpression is a novel leukemogenic mechanism that predicts adverse outcome in elderly patients with acute myeloid leukemia

3 Tables. 7 Figures. The online version of this article contains a data supplement.Acute myeloid leukemias (AMLs) result from multiple genetic alterations in hematopoietic stem cells. We describe a novel t(12;18)(p13;q12) involving ETV6 in a patient with AML. The translocation resulted in overexpression of SETBP1 (18q12), located close to the breakpoint. Overexpression of SETBP1 through retroviral insertion has been reported to confer growth advantage in hematopoietic progenitor cells. We show that SETBP1 overexpression protects SET from protease cleavage, increasing the amount of full-length SET protein and leading to the formation of a SETBP1–SET-PP2A complex that results in PP2A inhibition, promoting proliferation of the leukemic cells. The prevalence of SETBP1 overexpression in AML at diagnosis (n = 192) was 27.6% and was associated with unfavorable cytogenetic prognostic group, monosomy 7, and EVI1 overexpression (P < .01). Patients with SETBP1 overexpression had a significantly shorter overall survival, and the prognosis impact was remarkably poor in patients older than 60 years in both overall survival (P = .015) and event-free survival (P = .015). In summary, our data show a novel leukemogenic mechanism through SETBP1 overexpression; moreover, multivariate analysis confirms the negative prognostic impact of SETBP1 overexpression in AML, especially in elderly patients, where it could be used as a predictive factor in any future clinical trials with PP2A activators.This work was supported by the Ministerio de Educación y Ciencia, Ministerio de Ciencia e Innovación (PI081687, M.D.O.; and SAF2007-61827, C.B.), Departamento de Salud del Gobierno de Navarra (14/2008), ISCIII-RTICC (RD06/0020/0078), and Fundación para la Investigación Médica Aplicada y UTE (Spain).Elizabeth Guruceaga, of the Unit of Proteomics,Genomics and Bioinformatics of Center for Applied Medical Research, for the bioinformatics analysis, and Enrique Andreu for useful discussionPeer reviewe

The apoptosis-inducing factor family: Moonlig htingproteins in the crosstalk between mitochondria and nuclei

14 pags., 4 figs.In Homo sapiens, the apoptosis-inducing factor (AIF) family is represented bythree different proteins, known as AIF, AMID and AIFL, that have in commonthe mitochondrial localisation in healthy cells, the presence of FAD- andNADH-dependent domains involved in an -albeit yet not well understood- oxi-doreductase function and their capability to induce programmed cell death.AIF is the best characterised family member, while the information aboutAMID and AIFL is much scarcer. Nonetheless, available data support differentroles as well as mechanisms of action of their particular apoptogenic and redoxdomains regarding both pro-apoptotic and anti-apoptotic activities. Moreover,diverse cellular functions, to date far from fully clarified, are envisaged for thetranscripts corresponding to these three proteins. Here, we review the so faravailable knowledge on the moonlighting human AIF family from their molec-ular properties to their relevance in health and disease, through the evaluationof their potential cell death and redox functions in their different subcellularlocations. This picture emerging from the current knowledge of the AIF familyenvisages its contribution to regulate signalling and transcription machineriesin the crosstalk among mitochondria, the cytoplasm and the nucleusGovernment of Aragón-FEDER, Grant/Award Number: E35_20R; SpanishMinistry of Science and Innovation-StateResearch Agency (MICINN), Grant/Award Number: PID2019-103901GB-I00Peer reviewe

Pyridoxal 5′-Phosphate Biosynthesis by Pyridox-(am)-ine 5′-Phosphate Oxidase: Species-Specific Features

Enzymes reliant on pyridoxal 5′-phosphate (PLP), the metabolically active form of vitamin B6, hold significant importance in both biology and medicine. They facilitate various biochemical reactions, particularly in amino acid and neurotransmitter metabolisms. Vitamin B6 is absorbed by organisms in its non-phosphorylated form and phosphorylated within cells via pyridoxal kinase (PLK) and pyridox-(am)-ine 5′-phosphate oxidase (PNPOx). The flavin mononucleotide-dependent PNPOx enzyme converts pyridoxine 5′-phosphate and pyridoxamine 5′-phosphate into PLP. PNPOx is vital for both biosynthesis and salvage pathways in organisms producing B6 vitamers. However, for those depending on vitamin B6 as a nutrient, PNPOx participates only in the salvage pathway. Transferring the PLP produced via PNPOx to client apo-enzymes is indispensable for their catalytic function, proper folding and targeting of specific organelles. PNPOx activity deficiencies due to inborn errors lead to severe neurological pathologies, particularly neonatal epileptic encephalopathy. PNPOx maintains PLP homeostasis through highly regulated mechanisms, including structural alterations throughout the catalytic cycle and allosteric PLP binding, influencing substrate transformation at the active site. Elucidation at the molecular level of the mechanisms underlying PNPOx activity deficiencies is a requirement to develop personalized approaches to treat related disorders. Finally, despite shared features, the few PNPOx enzymes molecularly and functionally studied show species-specific regulatory properties that open the possibility of targeting it in pathogenic organisms

Los grandes olvidados del Reino Animal: el rompecabezas de los Invertebrados

En este trabajo se pretende integrar los conocimientos adquiridos en el máster y a través de la investigación de la literatura con la experiencia incomparable de los Prácticum, culminando en una serie de reflexiones sobre ambos períodos así como alguna propuesta de mejora.En primer lugar, se analizan desde un punto de vista crítico dos actividades realizadas en el máster: (1) Ideas alternativas en ciencias experimentales de la asignatura “Fundamentos de diseño instruccional y metodologías de aprendizaje en la especialidad de Física y Química y Biología y Geología” del 1er semestre; y (2) Práctica 5. Disección de órganos de la asignatura “Diseño, organización y desarrollo de actividades para el aprendizaje de Biología y Geología” del 2do semestre. El ensamblaje de ambas, tras una posterior adaptación al contexto, resultó en la propuesta didáctica que fue implementada a lo largo del Prácticum 3.En segundo lugar, se expone dicha propuesta didáctica en detalle, incorporando el análisis tanto de la literatura consultada previamente como de los resultados obtenidos. Aunque se explicará en profundidad en el apartado 5. Actividades, a continuación se resume brevemente: a. Contenido: se trata el tema de “Los Animales Invertebrados” de 1º de la E.S.O. b. Metodología: se emplea la técnica rompecabezas de Aronson et al. (1978), en combinación con el uso de Tecnologías de la Información y la Comunicación (TIC) c. Estructura: tiene lugar a lo largo de cuatro sesiones, una sesión previa de introducción, dos sesiones de desarrollo de la metodología escogida, y una sesión final de conclusiónPor último, se recogen varias conclusiones sobre el máster y los Prácticum, resaltando lo más provechoso, argumentando posibles puntos débiles y explorando algunas propuestas para su continuo proceso de mejora.<br /

SETBP1 overexpression is a novel leukemogenic mechanism that predicts adverse outcome in elderly patients with acute myeloid leukemia

3 Tables. 7 Figures. The online version of this article contains a data supplement.Acute myeloid leukemias (AMLs) result from multiple genetic alterations in hematopoietic stem cells. We describe a novel t(12;18)(p13;q12) involving ETV6 in a patient with AML. The translocation resulted in overexpression of SETBP1 (18q12), located close to the breakpoint. Overexpression of SETBP1 through retroviral insertion has been reported to confer growth advantage in hematopoietic progenitor cells. We show that SETBP1 overexpression protects SET from protease cleavage, increasing the amount of full-length SET protein and leading to the formation of a SETBP1–SET-PP2A complex that results in PP2A inhibition, promoting proliferation of the leukemic cells. The prevalence of SETBP1 overexpression in AML at diagnosis (n = 192) was 27.6% and was associated with unfavorable cytogenetic prognostic group, monosomy 7, and EVI1 overexpression (P < .01). Patients with SETBP1 overexpression had a significantly shorter overall survival, and the prognosis impact was remarkably poor in patients older than 60 years in both overall survival (P = .015) and event-free survival (P = .015). In summary, our data show a novel leukemogenic mechanism through SETBP1 overexpression; moreover, multivariate analysis confirms the negative prognostic impact of SETBP1 overexpression in AML, especially in elderly patients, where it could be used as a predictive factor in any future clinical trials with PP2A activators.This work was supported by the Ministerio de Educación y Ciencia, Ministerio de Ciencia e Innovación (PI081687, M.D.O.; and SAF2007-61827, C.B.), Departamento de Salud del Gobierno de Navarra (14/2008), ISCIII-RTICC (RD06/0020/0078), and Fundación para la Investigación Médica Aplicada y UTE (Spain).Elizabeth Guruceaga, of the Unit of Proteomics,Genomics and Bioinformatics of Center for Applied Medical Research, for the bioinformatics analysis, and Enrique Andreu for useful discussionPeer reviewe