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Puntos para una lectura rapida • Uno de los principales objetivos de la medicina maternofetal durante el seguimiento gestacional es la deteccion de anomalias fetales, tanto defectos morfologicos como funcionales o geneticos. • En los ultimos anos, un gran avance cientifico ha obligado a replantear la estrategia de cribado: la posibilidad de detectar ADN libre circulante en el plasma materno. • El desarrollo de tecnicas de secuenciacion masiva o de “nueva generacion” permite analizar este ADN fetal libre en la sangre materna. • Uno de los principales puntos a favor de la prueba del ADN fetal en sangre materna es su alto valor predictivo positivo (de hasta el 99% para la trisomia 21 frente al 4% del triple screening) y un valor predictivo negativo tambien muy alto, en torno al 99,9% para el sindrome de Down, y ligeramente inferior para las otras aneuploidias. • Es importante tambien tener en cuenta que la ecografia de primer trimestre, entre las 11,2 y las 14 semanas, aporta informacion amplia y permite el estudio anatomico precoz, asi como realizar cribados de riesgo para otras enfermedades, principalmente la preeclampsia, por lo que no parece recomendable dejar de realizarla. • En caso de que la prueba de ADN fetal informe de alto riesgo, este debera confirmarse mediante el analisis del cariotipo de la muestra obtenida mediante una prueba invasiva, ya sea biopsia corial o amniocentesis.

The aim of this work was to report the detection of a putative novel Ehrlichia strain associated with the tick Amblyomma triste. Free-living adult ticks determined as A. triste were collected by drag-sampling in Argentina and Uruguay. Molecular detection of Ehrlichia agents was performed targeting three different loci: 16S rRNA gene, dsb gene and a fragment of groESL heat shock operon. In total, 164 adults of A. triste (38 from INTA E.E.A Delta del Parana in Argentina and 126 from Toledo Chico in Uruguay) were analyzed. One tick (0.6%) collected in INTA E.E.A. Delta del Parana (Argentina) was positive. The phylogenetic analyses show that the Ehrlichia strain found in this study (named Ehrlichia sp. strain Delta) represents an independent lineage within the genus Ehrlichia, close to E. chaffeensis and E. muris. This is also the first report of an Ehrlichia agent infecting the tick A. triste. The medical and veterinary significance of Ehrlichia sp. strain Delta remains to be demonstrated. However, it is important to mention that adults of A. triste are aggressive to humans and domestic mammals. Therefore, the potential role of A. triste in the transmission of Ehrlichia agents to humans or domestic animals across its distributional range should be highlighted, even more considering that Ehrlichia sp. strain Delta is phylogenetically related to the zoonotic E. chaffeensis, which is recognized as pathogenic to both humans and animals.

Extracellular vesicles (EVs) are cell-derived vesicles important in intercellular communication that play an essential role in host-pathogen interactions, spreading pathogen-derived as well as host-derived molecules during infection. Pathogens can induce changes in the composition of EVs derived from the infected cells and use them to manipulate their microenvironment and, for instance, modulate innate and adaptive inflammatory immune responses, both in a stimulatory or suppressive manner. Gastric cancer is one of the leading causes of cancer-related deaths worldwide and infection with Helicobacter pylori (H. pylori) is considered the main risk factor for developing this disease, which is characterized by a strong inflammatory component. EVs released by host cells infected with H. pylori contribute significantly to inflammation, and in doing so promote the development of disease. Additionally, H. pylori liberates vesicles, called outer membrane vesicles (H. pylori-OMVs), which contribute to atrophia and cell transformation in the gastric epithelium. In this review, the participation of both EVs from cells infected with H. pylori and H. pylori-OMVs associated with the development of gastric cancer will be discussed. By deciphering which functions of these external vesicles during H. pylori infection benefit the host or the pathogen, novel treatment strategies may become available to prevent disease.

RESUMEN Objetivos Identificar las causas de accidentes y enfermedades profesionales de buzos mariscadores en las caletas de Coquimbo, Los Vilos y Tongoy en la región de Coquimbo (Chile), en el periodo 2008-2018. Métodos Se aplicó una encuesta de 44 preguntas para determinar las causas o factores que influyen en los accidentes y enfermedades profesionales de los buzos mariscadores en la región. Se recopiló información, experiencias y observaciones de los protagonistas. Para su análisis se utilizaron la tabla de doble entradas, los estadísticos de asociación X2 y C de contingencia. Resultados De las 52 encuestas realizadas, 28 buzos mariscadores han tenido accidentes y enfermedades profesionales (53%). El segmento que no completa la enseñanza básica concentra el 46% de los accidentes. También muestra incidencia, la profundidad a la cual se realiza el buceo, el consumo de alcohol y tabaco, el no uso de las tablas de descompresión y el no haber recibido capacitación. Conclusiones Los tipos de accidente o enfermedades profesionales asociadas de mayor incidencia son el síndrome de descompresión y la asfixia. Ambos casos presentan una alta asociación con el consumo de alcohol y el no uso de las tablas de descomprensión. ABSTRACT Objectives To identify the causes of accidents and occupational diseases of shell-fish divers in the small coves de Coquimbo, Los Vilos, and Tongoy, in the Coquimbo Region, Chile, in the period 2008-2018. Methods A structured survey of 44 questions was applied to determine the causes or factors that influence accidents and occupational diseases of shellfish divers in the region; collecting information, experiences and observations of the protagonists. The double entry table, the X2 and C contingency association statistics were used for the data analysis. Results Of the 52 surveys carried out, 28 shellfish divers have had accidents and occupational diseases (53%). The segment that does not complete basic education concentrates 46% of accidents. It also shows incidence, the depth at which diving is done, the consumption of alcohol and tobacco, the non-use of decompression tables and the lack of training. Conclusions The type of accident or associated occupational disease with the highest incidence is decompression syndrome and suffocation, both of which have a high association with alcohol consumption and non-use of decompression tables.

Funder: Instituto Nacional de Salud, Bogotá, Colombia, Salmonella Typhi (S. Typhi) is the causative agent of typhoid fever; a systemic disease affecting ~20 million people per year globally. There are little data regarding the contemporary epidemiology of typhoid in Latin America. Consequently, we aimed to describe some recent epidemiological aspects of typhoid in Colombia using cases reported to the National Public Health Surveillance System (Sivigila) between 2012 and 2015. Over the four-year reporting period there were 836 culture confirmed cases of typhoid in Colombia, with the majority (676/836; 80.1%) of reported cases originated from only seven departments. We further characterized 402 S. Typhi isolates with available corresponding data recovered from various departments of Colombia through antimicrobial susceptibility testing and molecular subtyping. The majority (235/402; 58.5%) of these typhoid cases occurred in males and were most commonly reported in those aged between 10 and 29 years (218/402; 54.2%); there were three (0.74%) reported fatalities. The overwhelming preponderance (339/402; 84.3%) of S. Typhi were susceptible to all tested antimicrobials. The most common antimicrobial to which the organisms exhibited non-susceptibility was ampicillin (30/402;7.5%), followed by nalidixic acid (23/402, 5.7%). Molecular subtyping identified substantial genetic diversity, which was well distributed across the country. Despite the diffuse pattern of S. Typhi genotypes, we identified various geographical hotspots of disease associated with local dominant genotypes. Notably, we found limited overlap of Colombian genotypes with organisms reported in other Latin American countries. Our work highlights a substantial burden of typhoid in Colombia, characterized by sustained transmission in some regions and limited epidemics in other departments. The disease is widely distributed across the country and associated with multiple antimicrobial susceptible genotypes that appear to be restricted to Colombia. This study provides a current perspective for typhoid in Latin America and highlights the importance of pathogen-specific surveillance to add insight into the limited epidemiology of typhoid in this region.

Genome editing has proven to be highly potent in the generation of functional gene knockouts in dividing cells. In the CNS however, efficient technologies to repair sequences are yet to materialize. Reprogramming on the mRNA level is an attractive alternative as it provides means to perform in situ editing of coding sequences without nuclease dependency. Furthermore, de novo sequences can be inserted without the requirement of homologous recombination. Such reprogramming would enable efficient editing in quiescent cells (e.g., neurons) with an attractive safety profile for translational therapies. In this study, we applied a novel molecular-barcoded screening assay to investigate RNAtrans-splicing in mammalian neurons. Through three alternative screening systems in cell culture and in vivo, we demonstrate that factors determiningtrans-splicing are reproducible regardless of the screening system. With this screening, we have located the most permissivetrans-splicing sequences targeting an intron in the Synapsin I gene. Using viral vectors, we were able to splice full-length fluorophores into the mRNA while retaining very low off-target expression. Furthermore, this approach also showed evidence of functionality in the mouse striatum. However, in its current form, thetrans-splicing events are stochastic and the overall activity lower than would be required for therapies targeting loss-of-function mutations. Nevertheless, the herein described barcode-based screening assay provides a unique possibility to screen and map large libraries in single animals or cell assays with very high precision.

Use of glyburide in gestational diabetes (GDM) has raised concerns about fetal and neonatal side effects, including increased birth weight. Placental nutrient transport is a key determinant of fetal growth, however the effect of glyburide on placental nutrient transporters is largely unknown. We hypothesized that glyburide treatment in GDM pregnancies is associated with increased expression of nutrient transporters in the syncytiotrophoblast plasma membranes. We collected placentas from GDM pregnancies who delivered at term and were treated with either diet modification (n = 15) or glyburide (n = 8). Syncytiotrophoblast microvillous (MVM) and basal (BM) plasma membranes were isolated and expression of glucose (glucose transporter 1; GLUT1), amino acid (sodium-coupled neutral amino acid transporter 2; SNAT2 and L-type amino acid transporter 1; LAT1) and fatty acid (fatty acid translocase; FAT/CD36, fatty acid transporter 2 and 4; FATP2, FATP4) transporters was determined by Western blot. Additionally, we determined GLUT1 expression by confocal microscopy in cultured primary human trophoblasts (PHT) after exposure to glyburide. Birth weight was higher in the glyburide-treated group as compared to diet-treated GDM women (3764 ± 126 g vs. 3386 ± 75 g; p 0.05). GLUT1 expression was increased in both MVM (+50%; p 0.01) and BM (+75%; p 0.01). In contrast, MVM FAT/CD36 (-65%; p = 0.01) and FATP2 (-65%; p = 0.02) protein expression was reduced in mothers treated with glyburide. Glyburide increased membrane expression of GLUT1 in a dose-dependent manner in cultured PHT. This data is the first to show that glyburide increases GLUT1 expression in syncytiotrophoblast MVM and BM in GDM pregnancies, and may promote transplacental glucose delivery contributing to fetal overgrowth.

Resumen Introduccion Salmonella spp. es un enteropatogeno que se transmite a los humanos a traves de alimentos o agua contaminada. En 1997, el Grupo de Microbiologia del Instituto Nacional de Salud de Colombia inicio el programa de vigilancia de enfermedad diarreica aguda y fiebre tifoidea, que incluye Salmonella spp. Este informe presenta los resultados fenotipicos y genotipicos de los aislamientos recuperados de 2005 a 2011 como parte de la vigilancia. Metodos Un total de 4.010 aislamientos de Salmonella spp. fueron analizados por serotipificacion con el esquema Kauffmann-White-LeMinor, patrones de sensibilidad antimicrobiana y de electroforesis en gel de campos pulsados (PFGE). Resultados Se identificaron un total de 93 serovares, con 9 predominantes, Typhimurium, Enteritidis, Typhi, Dublin, Panama, Derby, Braenderup, Saintpaul y Uganda. Salmonella spp. presento altos porcentajes de resistencia a tetraciclina y acido nalidixico. El 52,4% (2.101/4.010) de los aislamientos fueron sensibles a todos los antibioticos. La multirresistencia se observo en el 54,9% de los aislamientos de Typhimurium, representada por 81 combinaciones. Por PFGE se analizaron 51,9% aislamientos (2.083/4.010) de 34 serovares, generando 828 patrones electroforeticos XbaI. De estos, 8 se reportaron en al menos 2 paises en Latinoamerica. Conclusion La vigilancia de Salmonella spp. permite conocer la distribucion de los serovares, su resistencia y la identificacion de clones endemicos en Colombia, aportando bases para un tratamiento optimo en las infecciones generadas por este patogeno y en el diseno de programas para disminuir la dispersion de aislamientos multirresistentes.

Introduction Salmonella is an enteropathogen acquired through contaminated food or water. In Colombia, Salmonella spp. is included in the national surveillance of Acute Diarrhoeal Diseases and typhoid fever initiated in 1997. This report shows the phenotype and genotype results obtained from 2005 to 2011. Methods A total of 4010 isolates of Salmonella enterica were analysed by serotyping with Kauffmann–White–LeMinor, antimicrobial resistance patterns, and pulse-field gel electrophoresis (PFGE). Results A total of 93 serovars were identified, of which, Typhimurium, Enteritidis, Typhi, Dublin, Panama, Derby, Braenderup, Saintpaul, and Uganda were prominent. The highest levels of resistance were found for tetracycline and nalidixic acid. Susceptibility was observed in 52.4% (2101/4010) of the isolates. Multi-resistance was recorded in 54.9% of Typhimurium isolates, with 81 different combinations. Using PFGE, 51.9% (2083/4010) isolates were analysed in 34 serovars, and 828 electrophoretic patterns were obtained. From these, 8 patterns were found in at least two Latin-American countries. Conclusion The surveillance of Salmonella spp. provides information on the serovar distribution, antimicrobial resistance, and clonal distribution in Colombia, as well as information to treat this disease and control the spread of antimicrobial bacterial resistance.

Introduction Cytoreductive surgery plus intraperitoneal hyperthermic chemotherapy (HIPEC) has recently been established as the treatment of choice for selected patients with peritoneal carcinomatosis of colonic origin. Until recently, the simultaneous presence of peritoneal and hepatic dissemination has been considered a contraindication for surgery. The aim of this paper is to analyze the morbidity, mortality and survival of patients with simultaneous peritoneal and hepatic resection with HIPEC for peritoneal carcinomatosis secondary to colon cancer. Methods Between January 2010 and January 2015, 61 patients were operated on, 16 had simultaneous peritoneal and hepatic dissemination (group RH+), and 45 presented only peritoneal dissemination (group RH−). Results There were no differences between the groups in terms of demographic data, length of surgery and extension of peritoneal disease. Postoperative grade III–V complications were significantly higher in the RH+ group (56.3 vs 26.6%; P =0.032). For the whole group, mortality rate was 3.2% (two patients in group RH−, and none in group RH+). Patients with liver resection had a longer postoperative stay (14.4 vs 23.1 days) ( P =.027). Median overall survival was 33 months for RH−, and 36 for RH+ group. Median disease-free survival was 16 months for RH−, and 24 months for RH+ group. Conclusions Simultaneous peritoneal cytoreduction and hepatic resection resulted in a significantly higher Clavien grade III–V morbidity and a longer hospital stay, although the results are similar to other major abdominal interventions. The application of multimodal oncological and surgical treatment may obtain similar long-term survival results in both groups.

Resumen Introduccion La citorreduccion con quimioterapia intraperitoneal hipertermica (HIPEC) se ha postulado como tratamiento de eleccion en pacientes seleccionados con carcinomatosis peritoneal por cancer de colon. La presencia simultanea de diseminacion peritoneal y hepatica ha sido considerada una contraindicacion para esta cirugia. El objetivo del presente estudio es analizar la morbimortalidad y supervivencia de los pacientes con carcinomatosis peritoneal por cancer de colon, intervenidos mediante una citorreduccion peritoneal y reseccion hepatica simultanea con HIPEC. Metodos Entre enero de 2010 y enero de 2015 se intervinieron 61 pacientes, 45 pacientes con carcinomatosis peritoneal (grupo RH−) y 16 con carcinomatosis peritoneal y metastasis hepaticas (grupo RH+). Resultados No hubo diferencias significativas entre los 2 grupos en los datos demograficos, ASA, duracion de la intervencion, ni extension de la enfermedad peritoneal. Las complicaciones postoperatorias Clavien III-V fueron significativamente superiores en el grupo RH+ (56,3 vs. 26,6%; p = 0,03). La mortalidad global de la serie fue del 3,2% (2 pacientes en el grupo RH− y ninguno en el grupo RH+). Los pacientes con reseccion hepatica presentaron una estancia hospitalaria significativamente mas larga (14,4 vs. 23,1 dias; p = 0,027). La mediana estimada de supervivencia global fue de 33 meses para RH− y de 36 meses para RH+, y la de supervivencia libre de enfermedad fue de 16 meses para RH− y de 24 para RH+. Conclusiones La citorreduccion peritoneal con reseccion hepatica simultanea presenta una morbilidad postoperatoria y una estancia hospitalaria significativamente mayores, aunque las cifras son similares a las de otras cirugias abdominales mayores. La aplicacion de un tratamiento oncologico y quirurgico multimodal permite obtener resultados de supervivencia similares en ambos grupos.

// Aroa Baragano Raneros 1 , Alfredo Minguela Puras 2 , Ramon M. Rodriguez 1 , Enrique Colado 3 , Teresa Bernal 3 , Eduardo Anguita 4 , Adela Vasco Mogorron 2 , Alberto Chaparro Gil 4 , Jose Ramon Vidal-Castineira 1 , Leonardo Marquez-Kisinousky 1 , Paula Diaz Bulnes 1 , Amelia Martinez Marin 5 , Maria Carmen Garcia Garay 6 , Beatriz Suarez-Alvarez 1, * , Carlos Lopez-Larrea 1, * 1 Department of Immunology, Hospital Universitario Central de Asturias, Oviedo, Spain 2 Immunology Service, Instituto Murciano de Investigacion Biosanitaria (IMIB), Hospital Clinico Universitario Virgen de la Arrixaca, Murcia, Spain 3 Department of Hematology, Hospital Universitario Central de Asturias, Oviedo, Spain 4 Hematology Department, Hospital Clinico San Carlos, Instituto de Investigacion Sanitaria San Carlos (IdISSC), Department of Medicine, Universidad Complutense de Madrid (UCM), Madrid, Spain 5 Hematology Service, Hospital Clinico Universitario Virgen de la Arrixaca, Murcia, Spain 6 Hematology Service, Hospital General Universitario Santa Lucia, Cartagena, Murcia, Spain * These authors have contributed equally to this work Correspondence to: Carlos Lopez-Larrea, email: inmuno@hca.es Beatriz Suarez-Alvarez, email: bsuarez@hca.es Keywords: acute myeloid leukemia (AML), DNA methylation, NKG2DL, NKG2D, TIMP3 Received: September 16, 2016 Accepted: March 16, 2017 Published: March 29, 2017 ABSTRACT Acute myeloid leukemia (AML) is a disease with great morphological and genetic heterogeneity, which complicates its prognosis and treatment. The hypomethylating agents azacitidine (Vidaza ® , AZA) and decitabine (Dacogen ® , DAC) have been approved for the treatment of AML patients, but their mechanisms of action are poorly understood. Natural killer (NK) cells play an important role in the recognition of AML blasts through the interaction of the activating NKG2D receptor with its ligands (NKG2DL: MICA/B and ULBPs1-3). However, soluble NKG2DL (sNKG2DL) can be released from the cell surface, impairing immune recognition. Here, we examined whether hypomethylating agents modulate the release of sNKG2DL from AML cells. Results demonstrated that AZA- and DAC-treated AML cells reduce the release of sNKG2DL, preventing downregulation of NKG2D receptor on the cell surface and promoting immune recognition mediated by NKG2D-NKG2DL engagement. We show that the shedding of MICA, MICB and ULBP2 is inhibited by the increased expression of TIMP3, an ADAM17 inhibitor, after DAC treatment. The TIMP3 gene is highly methylated in AML cells lines and in AML patients (25.5%), in which it is significantly associated with an adverse cytogenetic prognosis of the disease. Overall, TIMP3 could be a target of the demethylating treatments in AML patients, leading to a decrease in MICA, MICB and ULBP2 shedding and the enhancement of the lytic activity of NK cells through the immune recognition mediated by the NKG2D receptor.

Macrophage activation and polarization are closely linked with metabolic rewiring, which is required to sustain their biological functions. These metabolic alterations allow the macrophages to adapt to the microenvironment changes associated with inflammation or tissue damage (hypoxia, nutrient imbalance, oxidative stress, etc.) and to fulfill their highly energy-demanding proinflammatory and anti-microbial functions. This response is integrated via metabolic sensors that coordinate these metabolic fluxes with their functional requirements. Here we review how the metabolic and phenotypic plasticity of macrophages are intrinsically connected with the hypoxia stress sensors and the unfolded protein response in the endoplasmic reticulum, and how these molecular pathways participate in the maladaptive polarization of macrophages in human pathology and chronic inflammation.

[EN] Introduction To characterize a carbapenem-resistant Enterobacter cloacae complex isolate recovered from a patient from Ukraine. Methods The isolate was sent to a regional reference laboratory for molecular characterization by whole genome sequencing. Susceptibility assays, carbapenemase identification, imipenem hydrolysis and clonality were performed. Results The isolate showed resistance or reduced susceptibility to all β-lactam agents tested. Genome analysis led to the identification of an NDM-1-producing E. cloacae complex strain that was assigned to a new multilocus sequence type, ST932. The blaNDM-1 enzyme was located in a conjugative IncX3 plasmid of ca. 50 kb. In addition, blaCMH-3, a recently described AmpC β-lactamase sequence, which has not previously been reported in Europe, was also detected and its genetic environment was studied. Conclusion To our knowledge, this is the first reported case in Europe of an E. cloacae complex strain that produces both blaNDM-1 and blaCMH-3., [ES] Introducción Caracterizar una cepa de Enterobacter cloacae complex resistente a los carbapenémicos detectado en un paciente procedente de Ucrania. Métodos Se envió la cepa a un laboratorio regional de referencia para proceder a su caracterización molecular mediante secuenciación del genoma completo. Se realizaron estudios de susceptibilidad, identificación de carbapenemasas, hidrólisis del imipenem y de clonalidad. Resultados El aislado mostró resistencia o sensibilidad reducida a todos los betalactámicos estudiados. El análisis molecular permitió la identificación de una cepa de E. cloacae complex productor de NDM-1 que se asignó a un nuevo secuenciotipo, ST932. Se localizó la enzima blaNDM-1 en plásmido conjugativo IncX3 de 50 kb. Además, se detectó la enzima blaCMH-3, una nueva betalactamasa de tipo AmpC descrita recientemente pero no detectada con anterioridad en Europa, llevándose a cabo el estudio de su entorno genético. Conclusión Según nuestros conocimientos, esta es la primera descripción una cepa de E. cloacae complex productora de blaNDM-1 junto con blaCMH-3 en Europa.

17β-estradiol is a neuronal survival factor against oxidative stress that triggers its protective effect even in the absence of classical estrogen receptors. The polymodal transient receptor potential vanilloid subtype 1 (TRPV1) channel has been proposed as a steroid receptor implied in tissue protection against oxidative damage. We show here that TRPV1 is sufficient condition for 17β-estradiol to enhance metabolic performance in injured cells. Specifically, in TRPV1 expressing cells, the application of 17β-estradiol within the first 3 h avoided H2O2-dependent mitochondrial depolarization and the activation of caspase 3/7 protecting against the irreversible damage triggered by H2O2. Furthermore, 17β-estradiol potentiates TRPV1 single channel activity associated with an increased open probability. This effect was not observed after the application of 17α-estradiol. We explored the TRPV1-Estrogen relationship also in primary culture of hippocampal-derived neurons and observed that 17β-estradiol cell protection against H2O2-induced damage was independent of estrogen receptors pathway activation, membrane started and stereospecific. These results support the role of TRPV1 as a 17β-estradiol-activated ionotropic membrane receptor coupling with mitochondrial function and cell survival., Graphical Abstract TRPV1 is sufficient condition for 17β-estradiol resistance against oxidative stress-induced cell death. The cell death induced by oxidative stress could be characterized by a kinetic model of three state: alive, vulnerable and dead. In particular, the transition from alive to vulnerable state involves the depolarization of mitochondrial membrane potential, and the evolution to cell death the activation of caspases. Only in TRPV1 expressing cells, the initial application of 17β-estradiol protects against the irreversible damage triggered by oxidative stress, by inhibition of the transition from alive to vulnerable state. This mechanism includes blocking oxidative stress-dependent mitochondrial depolarization and the activation of caspases 3/7. Furthermore, 17β-estradiol potentiates TRPV1 activity associated with an increased open probability. Despite the relationship between the activation of TRPV1 and the maintenance of the mitochondrial membrane potential should be clarified, it could include the increase of the calcium buffer capacity of the mitochondria and/or the overexpression of anti-apoptotic proteins such as the Bcl-2 family as previously reported. Finally, the 17β-estradiol cell protection was independent of estrogen receptors, and was membrane started and stereospecific. These results support the role of TRPV1 as a 17β-estradiol ionotropic receptor being critical to cell survival.

Graft-versus-host disease (GVHD) is the main complication after allogeneic hematopoietic stem cell transplantation. We previously unveiled a correlation between proportions of C-C motif chemokine receptor 7 (CCR7)+ T cells in the apheresis and the risk of developing GVHD. We wanted to evaluate in vivo whether apheresis with low proportion of CCR7+ cells or treatment with an anti-human CCR7 monoclonal antibody (mAb) were suitable strategies to prevent or treat acute GVHD in preclinical xenogeneic models. Therapeutic anti-CCR7 mAb was the most effective strategy in both prophylactic and therapeutic settings where antibody drastically reduced in vivo lymphoid organ infiltration of donor CCR7+ T cells, extended lifespan and solved clinical signs. The antibody neutralized in vitro migration of naive and central memory T cells toward CCR7 ligands and depleted target CCR7+ subsets through complement activation. Both mechanisms of action spared CCR7− subsets, including effector memory and effector memory CD45RA+ T cells which may mediate graft versus leukemia effect and immunity against infections. Accordingly, the numbers of donor CCR7+ T cells in the apheresis were not associated to cytomegalovirus reactivation or the recurrence of the underlying disease. These findings provide a promising new strategy to prevent and treat acute GVHD, a condition where new specific, safety and effective treatment is needed.

[Objectives] This study aimed to isolate and characterise extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL-E) isolates from animals and wastewater in Tunisia., [Methods] ESBL-E from wastewater (n = 123 samples), faeces of healthy animals (poultry, sheep, goats and calves) (n = 140) and raw milk from healthy cows (n = 42) and goats (n = 20) were investigated. Antimicrobial susceptibility was determined according to CLSI recommendations. The blaTEM, blaSHV, blaCTX-M and blaOXA-48 genes were analysed by PCR and sequencing. Phylogenetic groups were determined by PCR for Escherichia coli isolates. The clonality of E. coli and Klebsiella pneumoniae isolates was determined by XbaI-PFGE and MLST., [Results] A total of 81 E. coli, 20 K. pneumoniae, 4 Enterobacter cloacae, 1 Citrobacter freundii and 1 Citrobacter braakii were isolated. The blaCTX-M-1 and blaCTX-M-15 genes were predominant in E. coli and K. pneumoniae isolates. E. cloacae and C. braakii isolates harboured the blaSHV-12 gene. The C. freundii isolated from wastewater carried blaCTX-M-15, blaTEM-1 and blaOXA-204. E. coli isolates belonged to phylogroups A (37), B1 (25), B2 (7) and D (12). Seventy-eight E. coli isolates were typeable by PFGE and were classified into 34 pulsotypes. The K. pneumoniae isolates belonged to 11 pulsotypes. The E. coli isolates belonged to sequence types ST131, ST224, ST162, ST845, ST5204, ST69, ST141 and ST10. The K. pneumoniae isolates belonged to ST405, ST147, ST564, ST307, ST152, ST45, ST661 and ST1564., [Conclusion] This is the first report of O25b-B23-CTX-M-27-ST131 E. coli isolates and of C. freundii carrying blaCTX-M-15, blaTEM-1 and blaOXA-204 in Tunisia., This study was supported by the Tunisian Ministry of Higher Education and Scientific Research (Tunisia) and the University of Seville (Seville, Spain).

Relapse into drug use is a significant problem for people recovering from addiction. The ability that conditioned cues have to reinstate and reinvigorate drug-seeking is potentiated over time (incubation of seeking), posing an additional difficulty for maintaining abstinence. While the prefrontal cortex has been involved in the incubation phenomenon and the extracellular matrix, perineuronal nets (PNNs) in particular, may play a vital role in brain plasticity associated to drug relapse, there are no comparative analyses between different drug classes and natural reinforcers. Here, we compare the effects of early (1 day) and protracted (30 days) withdrawal from to cocaine, heroin and sucrose self-administration on the total density and density per intensity range of PNNs of different territories of the prefrontal cortex of male Lewis rats. Our results show that cocaine self-administration increases the density of PNNs in the dorsal prelimbic, infralimbic and ventral orbitofrontal cortices, while protracted withdrawal reversesthis effect in the dorsal prelimbic cortex. Also, heroin self-administration increases the density of PNNs in the infralimbic cortex and ventral orbitofrontal cortices, but this effect is lost after 30 days of withdrawal in the infralimbic cortex. Finally, the self-administration of sucrose-sweetened water or the protracted withdrawal from this powerful reinforcer does not affect any of the PNN parameters analysed. Our results show that two different drugs of abuse (but not a natural reward) with specific pharmacological and physiological actions, differentially modulate PNNs in specific areas of the rodent prefrontal cortex with potential implications for the incubation of seeking phenomenon.

19 p.-6 fig., In recent years, the macrophage colony-stimulating factor (M-CSF) and granulocyte-macrophage CSF (GM-CSF) cytokines have been identified as opposing regulators of the inflammatory program. However, the two cytokines are simultaneously present in the inflammatory milieu, and it is not clear how cells integrate these signals. In order to understand the regulatory networks associated with the GM/M-CSF signaling axis, we analyzed DNA methylation in human monocytes. Our results indicate that GM-CSF induces activation of the inflammatory program and extensive DNA methylation changes, while M-CSF-polarized cells are in a less differentiated state. This inflammatory program is mediated via JAK2 associated with the GM-CSF receptor and the downstream extracellular signal-regulated (ERK) signaling. However, PI3K signaling is associated with a negative regulatory loop of the inflammatory program and M-CSF autocrine signaling in GM-CSF-polarized monocytes. Our findings describe the regulatory networks associated with the GM/M-CSF signaling axis and how they contribute to the establishment of the inflammatory program associated with monocyte activation., This work was supported by grants from the Plan Nacional de I+D+I 2013– 2016 ISCIII (Institute of Health Carlos III; PI16/01318, PI17/01244, PI17/ 0119, PI16/1900, and PI19/00184); the Gobierno del Principado de Asturias; the PCTI-Plan de Ciencia, Tecnología e Innovación 2013-2017 (grant IDI/ 2018/144); FEDER ‘‘Funding Program of the European Union’’; the Red Española de Investigación Renal (REDinREN) (RD16/0009/0020, RD016/0009/002, and RD016/0009/001); the Agencia Estatal de Investigación (AEI) (ayuda Juan de la Cierva-Incorporaciόn; IJCI-2017-33347 to R.M.R.); and the Instituto de Salud Carlos III (Contratos Sara Borrell; CD16/00033 to C.H.). CIC bioGUNE support was provided by the Basque Department of Industry, Tourism and Trade (Etortek and Elkartek programs), the Innovation Technology Department of Bizkaia County, the CIBERehd Network, and Spanish MINECO, the Severo Ochoa Excellence Accreditation (SEV-2016-0644).

The risk of developing gastric cancer is strongly linked to Helicobacter pylori (H. pylori) infection. Alternatively, autophagy is a conserved response that is important in cellular homeostasis and provides protection against bacterial infections. Although H. pylori is typically considered an extracellular bacterium, several reports indicate that it internalizes, possibly to avoid exposure to antibiotics. Mechanisms by which H. pylori manipulates host cell autophagic processes remain unclear and, importantly, none of the available studies consider a role for the secreted H. pylori virulence factor gamma-glutamyltranspeptidase (HpGGT) in this context. Here, we identify HpGGT as a novel autophagy inhibitor in gastric cells. Our experiments revealed that deletion of HpGGT increased autophagic flux following H. pylori infection of AGS and GES-1 gastric cells. In AGS cells, HpGGT disrupted the late stages of autophagy by preventing degradation in lysosomes without affecting lysosomal acidification. Specifically, HpGGT impaired autophagic flux by disrupting lysosomal membrane integrity, which leads to a decrease in lysosomal cathepsin B activity. Moreover, HpGGT was necessary for efficient internalization of the bacteria into gastric cells. This important role of HpGGT in internalization together with the ability to inhibit autophagy posits HpGGT as a key virulence factor in the development of gastric cancer.

Tissue engineering is a promising strategy to promote heart regeneration after a myocardial infarction (MI). In this study, we investigated the reparative potential of a system that combines adipose-derived stem cells (ADSCs) with microparticles (MPs) loaded with neuregulin (NRG), named ADSC-NRG-MPs, on a rat MI model. First, cells were attached to the surface of MPs encapsulating NRG and coated with a 1:1 mixture of collagen and poly-d-lysine. One week after in vivo administration, the system favored the shift of macrophage expression from a pro-inflammatory to a regenerative phenotype. At long-term, the adhesion of ADSCs to MPs resulted in an increased cell engraftment, with cells being detectable in the tissue up to three months. In consonance, better tissue repair was observed in the animals treated with cells attached to MPs, which presented thicker left ventricles than the animals treated with ADSCs alone. Moreover, the presence of NRG in the system promoted a more complete regeneration, reducing the infarct size and stimulating cardiomyocyte proliferation. Regarding vasculogenesis, the presence of ADSCs and NRG-MPs alone stimulated vessel formation when compared to the control group, but the combination of both induced the largest vasculogenic effect, promoting the formation of both arterioles and capillaries. Importantly, only when ADSCs were administered adhered to MPs, they were incorporated into newly formed vessels. Collectively, these findings demonstrate that the combination of ADSCs, MPs and NRG favored a synergy for inducing a greater and more complete improvement in heart regeneration and provided strong evidence to move forward with preclinical studies with this strategy.

The Recreovia program provides free physical activity (PA) classes in public spaces in Bogota, Colombia. The purpose of this study was to assess the effectiveness of the Recreovia program in increasing PA among users of nine parks in Bogota. This study was a natural experiment conducted between 2013 and 2015 in Bogota. Community members and park users living nearby three groups of parks were compared: Group 1 were parks implementing new Recreovias (n=3), Group 2 were control parks (n=3) without Recreovias, and Group 3 were parks with existing Recreovias. Individuals in the "intervention" group were exposed to newly implemented Recreovia programs in parks near their homes. Measurements were collected at baseline and 6-8months after the intervention started. A total of 1533 participants were enrolled in the study: 501 for the existing Recreovias (included in a cross-sectional assessment) and 1032 participants (from the new Recreovias and control parks) included in the cross-sectional and pre-post study. Most participants were low income females. Twenty-three percent of the intervention group started participating in the program. Users of existing Recreovias were significantly more active and less likely to be overweight/obese compared to new Recreovia users at baseline. No changes on PA were found when comparing the intervention and control groups. Recreovias may have potential for increasing PA at the population level in urban areas given their rapid scalability, the higher levels of PA observed among program users, and its potential to reach women, low-income, less educated populations, and the overweight and obese.

Multi-sectoral strategies to promote active recreation and physical activity in public spaces are crucial to building a “culture of health”. However, studies on the sustainability and scalability of these strategies are limited. This paper identifies the factors related to the sustainability and scaling up of two community-based programs offering physical activity classes in public spaces in Colombia: Bogota's Recreovia and Colombia's “Healthy Habits and Lifestyles Program- HEVS ”. Both programs have been sustained for more than 10 years, and have benefited 1455 communities. We used a mixed-methods approach including semi-structured interviews, document review and an analysis of data regarding the programs' history, characteristics, funding, capacity building and challenges. Interviews were conducted between May–October 2015. Based on the sustainability frameworks of Shediac-Rizkallah and Bone and Scheirer, we developed categories to independently code each interview. All information was independently analyzed by four of the authors and cross-compared between programs. Findings showed that these programs underwent adaptation processes to address the challenges that threatened their continuation and growth. The primary strategies included flexibility/adaptability, investing in the working conditions and training of instructors, allocating public funds and requesting accountability, diversifying resources, having community support and champions at different levels and positions, and carrying out continuous advocacy to include physical activity in public policies. Recreovia and HEVS illustrate sustainability as an incremental, multi-level process at different levels. Lessons learned for similar initiatives include the importance of individual actions and small events, a willingness to start small while dreaming big, being flexible, and prioritizing the human factor.

Human chorionic gonadotropin (hCG) is a key autocrine/paracrine regulator of placental syncytiotrophoblast, the transport epithelium of the human placenta. Syncytiotrophoblast hCG secretion is modulated by the partial pressure of oxygen (pO2), reactive oxygen species (ROS) and potassium (K+) channels. Here we test the hypothesis that K+ channels mediate the effects of pO2 and ROS on hCG secretion. Placental villous explants from normal term pregnancies were cultured for 6 days at 6% (normoxia), 21% (hyperoxia) or 1% (hypoxia) pO2. On days 3-5, explants were treated with 5mM 4-aminopyridine (4-AP) or tetraethylammonium (TEA), blockers of pO2-sensitive voltage-gated K+ (KV) channels, or ROS (10-1000μM H2O2). hCG secretion and lactate dehydrogenase (LDH) release, a marker of necrosis, were determined daily. At day 6, hCG and LDH were measured in tissue lysate and 86Rb (K+) efflux assessed to estimate syncytiotrophoblast K+ permeability. hCG secretion and 86Rb efflux were significantly greater in explants maintained in 21% pO2 than normoxia. 4-AP/TEA inhibited hCG secretion to a greater extent at 21% than 6% and 1% pO2, and reduced 86Rb efflux at 21% but not 6% pO2. LDH release and tissue LDH/hCG were similar in 6%, 21% and 1% pO2 and unaffected by 4-AP/TEA. H2O2 stimulated 86Rb efflux and hCG secretion at normoxia but decreased 86Rb efflux, without affecting hCG secretion, at 21% pO2. 4-AP/TEA-sensitive K+ channels participate in pO2-sensitive hCG secretion from syncytiotrophoblast. ROS effects on both hCG secretion and 86Rb efflux are pO2-dependent but causal links between the two remain to be established.

Although the importance of DNA methylation-dependent gene expression to neuronal plasticity is well established, the dynamics of methylation and demethylation during the induction and expression of synaptic plasticity have not been explored. Here, we combined electrophysiological, pharmacological, molecular, and immunohistochemical approaches to examine the contribution of DNA methylation and the phosphorylation of Methyl-CpG-binding protein 2 (MeCP2) to synaptic plasticity. We found that, at twenty minutes after theta burst stimulation (TBS), the DNA methylation inhibitor 5-aza-2-deoxycytidine (5AZA) impaired hippocampal long-term potentiation (LTP). Surprisingly, after two hours of TBS, when LTP had become a transcription-dependent process, 5AZA treatment had no effect. By comparing these results to those in naive slices, we found that, at two hours after TBS, an intergenic region of the RLN gene was hypomethylated and that the phosphorylation of residue S80 of MeCP2 was decreased, while the phosphorylation of residue S421 was increased. As expected, 5AZA affected only the methylation of the RLN gene and exerted no effect on MeCP2 phosphorylation patterns. In summary, our data suggest that tetanic stimulation induces critical changes in synaptic plasticity that affects both DNA methylation and the phosphorylation of MeCP2. These data also suggest that early alterations in DNA methylation are sufficient to impair the full expression of LTP.

Using a non-experimental, transverse, co-relational design, we assess the relationship between optimism and quality of life (QoL) in adults aged between 18 and 65 years. Two questionnaires were administered: WHOQOLBREF, which assessed QoL, and an instrument that assessed trait–state optimism. A total of 1190 subjects from various public and private high schools, institutions and commercial centers in the city of Antofagasta, Chile participated. The mean sample age was 40.0 years and 50.0% of respondents were male. There is a relationship between QoL and state optimism, with males demonstrating a higher QoL. Differences in optimism were observed only in age groups in which younger persons scored less than older persons. The results obtained indicate that there is a partial relationship between QoL and optimism, with state optimism having a significant effect on QoL.

Workshops are an important part of the IFPA annual meeting. At IFPA Meeting 2010 there were twelve themed workshops, six of which are summarized in this report. 1. The immunology workshop focused on normal and pathological functions of the maternal immune system in pregnancy. 2. The transport workshop dealt with regulation of ion and water transport across the syncytiotrophoblast of human placenta. 3. The epigenetics workshop covered DNA methylation and its potential role in regulating gene expression in placental development and disease. 4. The vascular reactivity workshop concentrated on methodological approaches used to study placental vascular function. 5. The workshop on epitheliochorial placentation covered current advances from in vivo and in vitro studies of different domestic species. 6. The proteomics workshop focused on a variety of techniques and procedures necessary for proteomic analysis and how they may be implemented for placental research.