TBL1 is required for the mesenchymal phenotype of transformed breast cancer cells

The epithelial-to-mesenchymal transition (EMT) and its reversion (MET) are related to tumor cell dissemination and migration, tumor circulating cell generation, cancer stem cells, chemoresistance, and metastasis formation. To identify chromatin and epigenetic factors possibly involved in the process of EMT, we compare the levels of expression of epigenetic genes in a transformed human breast epithelial cell line (HMEC-RAS) versus a stable clone of the same cell line expressing the EMT master regulator ZEB1 (HMEC-RAS-ZEB1). One of the factors strongly induced in the HMEC-RAS-ZEB1 cells was Transducin beta-like 1 (TBL1), a component of the NCoR complex, which has both corepressor and coactivator activities. We show that TBL1 interacts with ZEB1 and that both factors cooperate to repress the promoter of the epithelial gene E-cadherin (CDH1) and to autoactivate the ZEB1 promoter. Consistent with its central role, TBL1 is required for mesenchymal phenotypes of transformed breast epithelial and breast cancer cell lines of the claudin-low subtype. Importantly, a high expression of the TBL1 gene correlates with poor prognosis and increased proportion of metastasis in breast cancer patients, indicating that the level of TBL1 expression can be used as a prognostic marker.Ministerio de Economía y Competitividad BFU2014-53543-P, BFU2017-85420-RJunta de Andalucía BIO-32

MRGBP, a member of the NuA4 complex, inhibits DNA double-strand break repair

The repair of DNA breaks takes place in the context of chromatin and thus involves the activity of chromatin remodelers. The nucleosome acetyltransferase of H4 (NuA4) remodeler complex enables DNA break repair by relaxing flanking chromatin. Here, we show that MRG domain binding protein (MRGBP), a member of this complex, acts as a general inhibitor of DNA double-strand break repair. Upon its downregulation, repair is generally increased. This is particularly evident for the stimulation of early events of homologous recombination. Thus, MRGBP has an opposing role to the main catalytic subunits of the NuA4 complex. Our data suggest that MRGBP acts by limiting the activity of this complex in DNA repair, specifically by narrowing the extent of DNA-end resection.Ministerio de Economía y Competitividad SAF2016-74855-

Golgi localisation of GMAP210 requires two distinct cis-membrane binding mechanisms

<p>Abstract</p> <p>Background</p> <p>The Golgi apparatus in mammals appears as a ribbon made up of interconnected stacks of flattened cisternae that is positioned close to the centrosome in a microtubule-dependent manner. How this organisation is achieved and retained is not well understood. GMAP210 is a long coiled-coil cis-Golgi associated protein that plays a role in maintaining Golgi ribbon integrity and position and contributes to the formation of the primary cilium. An amphipathic alpha-helix able to bind liposomes <it>in vitro </it>has been recently identified at the first 38 amino acids of the protein (amphipathic lipid-packing sensor motif), and an ARF1-binding domain (Grip-related Arf-binding domain) was found at the C-terminus. To which type of membranes these two GMAP210 regions bind <it>in vivo </it>and how this contributes to GMAP210 localisation and function remains to be investigated.</p> <p>Results</p> <p>By using truncated as well as chimeric mutants and videomicroscopy we found that both the N-terminus and the C-terminus of GMAP210 are targeted to the cis-Golgi <it>in vivo</it>. The ALPS motif was identified as the N-terminal binding motif and appeared concentrated in the periphery of Golgi elements and between Golgi stacks. On the contrary, the C-terminal domain appeared uniformly distributed in the cis-cisternae of the Golgi apparatus. Strikingly, the two ends of the protein also behave differently in response to the drug Brefeldin A. The N-terminal domain redistributed to the endoplasmic reticulum (ER) exit sites, as does the full-length protein, whereas the C-terminal domain rapidly dissociated from the Golgi apparatus to the cytosol. Mutants comprising the full-length protein but lacking one of the terminal motifs also associated with the cis-Golgi with distribution patterns similar to those of the corresponding terminal end whereas a mutant consisting in fused N- and C-terminal ends exhibits identical localisation as the endogenous protein.</p> <p>Conclusion</p> <p>We conclude that the Golgi localisation of GMAP210 is the result of the combined action of the two N- and C-terminal domains that recognise different sub-regions of the cis-GA. Based on present and previous data, we propose a model in which GMAP210 would participate in homotypic fusion of cis-cisternae by anchoring the surface of cisternae via its C-terminus and projecting its distal N-terminus to bind the rims or to stabilise tubular structures connecting neighbouring cis-cisternae.</p

Perceptions on the management, exposure, biosafety and handling of cytostatics in the nursing staff of a private health institution in the Autonomous City of Buenos Aires

Artículo[ES] Introducción: El profesional de enfermería oncológica es formado con conocimientos sobre el cáncer, modalidades de tratamiento, escenarios de cuidado del paciente. El riesgo a la exposición resultante de la manipulación de los citostáticos puede afectar la salud de los agentes sanitarios. Objetivo: describir las medidas de bioseguridad para la manipulación de citostáticos y los signos clínicos y síntomas producto de la exposición a estos medicamentos en el personal de enfermería de una institución de salud privada de la Ciudad Autónoma de Buenos Aires. Métodos: Se realizó un estudio observacional, descriptivo, de corte transversal, retrospectivo. La muestra estuvo conformada por 31 profesionales de enfermería. Como instrumento recolección de datos se utilizó el propuesto por Villa y Varela-Díaz (2020), este instrumento registra datos sociodemográficos, laborales, de salud y sobre medidas de bioseguridad. Resultados: La muestra estuvo confirmada por un 74,2% del sexo femenino, la edad promedio fue de 37,67±6,79, el 58,1% era Licenciado/a en Enfermería, con una experiencia promedio en oncología de 4,06±4,09. El 96,8 % de los participantes administraba citostáticos y el 51,6 % participaba en el desecho. Los principales síntomas reportados fueron la cefalea y el dolor abdominal con 64,5% y 45,2% respectivamente. El 41,9% refiere la realización de exámenes paraclínicos y control por parte de la institución. Conclusiones: El personal de enfermería está expuesto a altos riesgos laborales cuando se trata de citostáticos. Se requiere el cumplimiento de los protocolos el manejo y descarte, con la finalidad de elevar los estándares de seguridad del paciente y seguridad laboral. [EN] Introduction: The oncology nurse practitioner is trained with knowledge about cancer, treatment modalities, patient care scenarios. The risk to exposure resulting from handling cytostatics can affect the health of healthcare workers. Objective: to describe the biosafety measures for handling cytostatics and the clinical signs and symptoms resulting from exposure to these drugs in the nursing staff of a private health care institution in the Autonomous City of Buenos Aires. Methods: An observational, descriptive, retrospective, cross-sectional, retrospective study was carried out. The sample consisted of 31 nursing professionals. The data collection instrument used was the one proposed by Villa and Varela-Díaz (2020), which records sociodemographic, occupational, health and biosafety measures data. Results: The sample was 74.2% female, the average age was 37.67±6.79, 58.1% had a Bachelor’s degree in Nursing, with an average experience in oncology of 4.06±4.09. Of the participants, 96.8% administered cytostatics and 51.6% were involved in disposal. The main symptoms reported were headache and abdominal pain with 64.5% and 45.2% respectively. 41.9% referred to paraclinical examinations and control by the institution. Conclusions: Nursing personnel are exposed to high occupational risks when dealing with cytostatics. Compliance with protocols for handling and disposal is required in order to raise patient safety and occupational safety standards.S

Phytochrome b nuclear bodies respond to the low red to far-red ratio and to the reduced irradiance of canopy shade in Arabidopsis

The current consensus is that plant responses to canopy shade involve the perception of low red to far-red ratios (R:FRs) by phytochrome B (phyB), which leads to the direct activation of auxin synthesis genes by PHYTOCHROME INTERACTING FACTORs (PIFs). In addition to its effect on R:FRs, shade also reduces irradiance, but whether shade-induced drops in irradiance affect phyB activity has not been demonstrated. To address this issue, we investigated whether irradiance and R:FRs have similar effects on the nuclear distribution of phyB in petiole cells of light-grown plants. Under high-irradiance white light, phyB formed large nuclear bodies. Lowering irradiance without changing R:FRs or lowering R:FRs by adding far-red light led to the appearance of small nuclear bodies containing phyB. Large nuclear bodies remained but with some concomitant reduction in diameter. The appearance of small nuclear bodies was rapid, stable, and reversible upon the return to high irradiance and high R:FRs. High levels of red light but not of blue light were enough to restrain the formation of small phyB nuclear bodies. Irradiance was effective within the range found in natural canopies and even under relatively low R:FRs. The promotion of leaf hyponasty by lowering irradiance was impaired in phyB and pif mutants, as previously reported for the response to R:FRs. The expression of auxin-related genes showed a similar hierarchy of response to low R:FRs and low irradiance. We propose that phyB is able to perceive not only the low R:FRs, but also the low irradiance of shade.Fil: Trupkin, Santiago Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; ArgentinaFil: Legris, Martina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Buchovsky, Ana Sabrina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; ArgentinaFil: Tolava Rivero, María Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; ArgentinaFil: Casal, Jorge Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones Fisiológicas y Ecológicas Vinculadas a la Agricultura; Argentin

Inhibiting arginine methylation as a tool to investigate cross-talk with methylation and acetylation post-translational modifications in a glioblastoma cell line

Glioblastomas (GBM) are the most common grade 4 brain tumours; patients have very poor prognosis with an average survival of 15 months after diagnosis. Novel research lines have begun to explore aberrant protein arginine methylation (ArgMe) as a possible therapeutic target in GBM and ArgMe inhibitors are currently in clinical trials. Enzymes known as protein arginine methyltransferases (PRMT1-9) can lead to mono-or di-ArgMe, and in the latter case symmetric or asymmetric dimethylation (SDMA and ADMA, respectively). Using the most common GBM cell line, we have profiled the expression of PRMTs, used ArgMe inhibitors as tools to investigate post-translational modifications cross-talk and measured the effect of ArgMe inhibitors on cell viability. We have identified novel SDMA events upon inhibition of ADMA in GBM cells and spheroids. We have observed cross-talk between ADMA and lysine acetylation in GBM cells and platelets. Treatment of GBM cells with furamidine, a PRMT1 inhibitor, reduces cell viability in 2D and 3D models. These data provide new molecular understanding of a disease with unmet clinical needs

Cambios inducidos por la radiación UV en películas de carboximetilcelulosa y alcohol polivinílico en presencia de benzoato de sodio

La carboximetilcelulosa sódica (CMC) es un derivado de la celulosa con propiedades filmogénicas, utilizada en el área de alimentos. El alcohol polivinílico (PVOH) es un polímero sintético semicristalino obtenido mediante hidrólisis del acetato de polivinilo. El foto entrecruzamiento de las películas con UV constituye una tecnología que permite mejorar las propiedades de los materiales. Para este tratamiento es necesario incorporar durante el procesamiento de la matriz fotoiniciadores como el benzoato de sodio (BS), para producir radicales que inicien las reacciones de reticulación. Además el BS funcionaliza el material otorgándole propiedades antimicrobianas. Los objetivos del trabajo fueron: i) desarrollar películas puras y compuestas a base de PVOH y CMC, estudiando sus propiedades fisicoquímicas, mecánicas, térmicas y micro estructurales; n) analizar las interacciones establecidas entre CMC y PVOH y los cambios inducidos por el tratamiento con radiación UV en presencia de benzoato de sodio (BS); ni) evaluar la capacidad antimicrobiana de las películas activas. Análisis térmicos, rayos X, SEM y FTIR evidenciaron los cambios micro estructurales producido en la matriz y proporcionaron información acerca de la modificación inducida ya sea por la presencia de BS o por el tratamiento con UV. Los espectros de FTIR reflejaron cierto grado de interacción polímero-polímero a nivel molecular en las regiones amorfas, especialmente en las películas compuestas fotoentrecruzadas con BS. La incorporación de este compuesto activo combinado con UV fue positiva debido a la inhibición del crecimiento de un amplio espectro de microorganismos (E. coli, Salmonella spp, Penicilium sp. y Canclicla spp.). Asimismo, desde el punto de vista físico-químico y estructural el tratamiento de las películas compuestas modificó significativamente su morfología evidenciada por SEM, obteniendo materiales más insolubles en agua. El desarrollo de películas funcionalizadas permitió obtener un material potencialmente aplicable en el envasado de alimentos.Facultad de Ciencias ExactasCentro de Investigación y Desarrollo en Criotecnología de AlimentosFacultad de Ingenierí

Trypanosomatid-caused conditions: State of the art of therapeutics and potential applications of lipid-based nanocarriers

Trypanosomatid-caused conditions (African trypanosomiasis, Chagas disease, and leishmaniasis) are neglected tropical infectious diseases that mainly affect socioeconomically vulnerable populations. The available therapeutics display substantial limitations, among them limited efficacy, safety issues, drug resistance, and, in some cases, inconvenient routes of administration, which made the scenarios with insufficient health infrastructure settings inconvenient. Pharmaceutical nanocarriers may provide solutions to some of these obstacles, improving the efficacy–safety balance and tolerability to therapeutic interventions. Here, we overview the state of the art of therapeutics for trypanosomatid-caused diseases (including approved drugs and drugs undergoing clinical trials) and the literature on nanolipid pharmaceutical carriers encapsulating approved and non-approved drugs for these diseases. Numerous studies have focused on the obtention and preclinical assessment of lipid nanocarriers, particularly those addressing the two currently most challenging trypanosomatid-caused diseases, Chagas disease, and leishmaniasis. In general, in vitro and in vivo studies suggest that delivering the drugs using such type of nanocarriers could improve the efficacy–safety balance, diminishing cytotoxicity and organ toxicity, especially in leishmaniasis. This constitutes a very relevant outcome, as it opens the possibility to extended treatment regimens and improved compliance. Despite these advances, last-generation nanosystems, such as targeted nanocarriers and hybrid systems, have still not been extensively explored in the field of trypanosomatid-caused conditions and represent promising opportunities for future developments. The potential use of nanotechnology in extended, well-tolerated drug regimens is particularly interesting in the light of recent descriptions of quiescent/dormant stages of Leishmania and Trypanosoma cruzi, which have been linked to therapeutic failure.Fil: Muraca, Giuliana Sabrina. Universidad Nacional de La Plata. Facultad de Ciencas Exactas. Laboratorio de Investigación y Desarrollo de Bioactivos; Argentina. Ministerio de Salud. Administración Nacional de Medicamentos, Alimentos y Tecnología Médica; ArgentinaFil: Rivero Berti, Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; ArgentinaFil: Sbaraglini, Maria Laura. Universidad Nacional de La Plata. Facultad de Ciencas Exactas. Laboratorio de Investigación y Desarrollo de Bioactivos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Fávaro, Wagner J.. Universidade Estadual Do Campinas. Instituto de Biologia. Departamento de Biologia Estructural y Funcional.; BrasilFil: Durán, Nelson. Universidade Estadual Do Campinas. Instituto de Biologia. Departamento de Biologia Estructural y Funcional.; Brasil. Universidad Federal do Abc; BrasilFil: Castro, Guillermo Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Fermentaciones Industriales. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Fermentaciones Industriales; ArgentinaFil: Talevi, Alan. Universidad Nacional de La Plata. Facultad de Ciencas Exactas. Laboratorio de Investigación y Desarrollo de Bioactivos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentin

Differential β₂-adrenergic receptor expression defines the phenotype of non-tumorigenic and malignant human breast cell lines

Breast cancer is the most frequent malignancy in women. Several reports demonstrated that adrenergic receptors (ARs) are involved in breast cancer. Here we observed that epinephrine (Epi), an endogenous AR agonist, caused opposite effects in non-tumorigenic (MCF-10A and HBL-100) and tumor cells (MCF-7 and MDA-MB-231). Thus, Epi, in non-tumor breast cells, as well as isoproterenol (β-agonist), in all cell lines, maintained a benign phenotype, decreasing cell proliferation and migration, and stimulating cell adhesion. β-AR expression and cAMP levels were higher in MCF-10A than in MCF-7 cells. β₂-AR knock-down caused a significant increase of cell proliferation and migration, and a decrease of cell adhesion both in basal and in Iso-stimulated conditions. Coincidently, β₂-AR over-expression induced a significant decrease of cell proliferation and migration, and an increase of cell adhesion. Therefore, β₂-AR is implied in cell phenotype and its agonists or antagonists could eventually complement cancer therapy.Fil: Gargiulo, Lucía. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Copsel, Sabrina Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Farmacología; ArgentinaFil: Rivero, Ezequiel Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Galés, Céline. Inserm; FranciaFil: Sénard, Jean Michel. Inserm; FranciaFil: Luthy, Isabel Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Davio, Carlos Alberto. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Farmacología; ArgentinaFil: Bruzzone, Ariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentin

Serological and molecular survey of hepatitis E virus in cats and dogs in Spain

Hepatitis E virus (HEV) is an emerging zoonotic pathogen that is currently recognized as one of themajor causes of acute human hepatitis worldwide. In Europe, the increasing number of hepatitis E cases is mainly associated with the consumption of animal food products or contact with infected animals. Dogs and cats have been suggested as a zoonotic source of HEV infection. The aim of this study was to assess Orthohepevirus circulation, including HEV-A, HEV-B and HEV-C species, in sympatric urban cats and dogs in southern Spain. Between 2017 and 2020, blood samples were collected from 144 stray cats and 152 dogs, both strays and pets. The presence of antibodies againstHEV were tested using a double-antigen sandwich ELISA and seropositive simples were further analysed bywestern blot.ART-PCR was performed to detect RNAof Orthohepevirus species (HEV-A,HEV-B andHEV-C).Atotal of 19 (6.4%; 95%CI: 3.6-9.2) of the 296 animals tested showed anti-HEV antibodies by ELISA. Seropositivity was significantly higher in dogs (9.9%; 15/152; 95%CI: 5.1-14.6) than in cats (2.8%; 4/144; 95%CI: 0.1-5.5). Ten of the 18 ELISA-positive animals that could be further analysed by western blot, reacted against HEV-3 and/or HEV-C1 antigens, which suggest circulation of both genotypes in urban cats and dogs in the study area. However, HEV-A, HEV-B and HEV-C RNA were not detected in any of the tested sera. This is the first study to assess HEV circulation in both stray cats and dogs in Europe. Our results provide evidence of HEV exposure in sympatric urban cat and dog populations in southern Spain. Further studies are needed to determine the role of these species in the epidemiology of HEV