Wnt/β-Catenin Signaling Enhances Cyclooxygenase-2 (COX2) Transcriptional Activity in Gastric Cancer Cells

BACKGROUND: Increased expression of the cyclooxygenase-2 enzyme (COX2) is one of the main characteristics of gastric cancer (GC), which is a leading cause of death in the world, particularly in Asia and South America. Although the Wnt/β-catenin signaling pathway has been involved in the transcriptional activation of the COX2 gene, the precise mechanism modulating this response is still unknown. METHODOLOGY/PRINCIPAL FINDINGS: Here we studied the transcriptional regulation of the COX2 gene in GC cell lines and assessed whether this phenomenon is modulated by Wnt/β-catenin signaling. We first examined the expression of COX2 mRNA in GC cells and found that there is a differential expression pattern consistent with high levels of nuclear-localized β-catenin. Pharmacological treatment with either lithium or valproic acid and molecular induction with purified canonical Wnt3a significantly enhanced COX2 mRNA expression in a dose- and time-dependent manner. Serial deletion of a 1.6 Kbp COX2 promoter fragment and gain- or loss-of-function experiments allowed us to identify a minimal Wnt/β-catenin responsive region consisting of 0.8 Kbp of the COX2 promoter (pCOX2-0.8), which showed maximal response in gene-reporter assays. The activity of this pCOX2-0.8 promoter region was further confirmed by site-directed mutagenesis and DNA-protein binding assays. CONCLUSIONS/SIGNIFICANCE: We conclude that the pCOX2-0.8 minimal promoter contains a novel functional T-cell factor/lymphoid enhancer factor (TCF/LEF)-response element (TBE Site II; -689/-684) that responds directly to enhanced Wnt/β-catenin signaling and which may be important for the onset/progression of GC

La extensión al servicio de la enseñanza del ranquel

En este artículo se expone el desarrollo del trabajo de extensión realizado con los ranqueles, pueblo originario de la región pampeana, en la revitalización de su lengua ancestral. Para ello nos referiremos a los estudios sobre el ranquel y al cambio operado en los grupos indígenas en lo que hace a las actitudes hacia su propia lengua a partir del año 1994, en que la Reforma Constitucional les otorga diferentes derechos, entre ellos, el derecho a aprender a leer y a escribir en su lengua nativa. A partir de entonces, y a pedido de los ranqueles, el Ministerio de Cultura y Educación de La Pampa nombra dos docentes pertenecientes a este grupo étnico quienes, a partir de 1996, dictan cursos de ranquel en distintos lugares de la provincia. Sin embargo, no existían materiales didácticos que los ayudaran en el aula. Ante esta situación, en 2006 se inician contactos con los maestros ranqueles que dan por resultado la implementación de un proyecto que se desarrolla entre 2010 y 2012. En el año 2013 se presenta el segundo proyecto que actualmente (agosto, 2016) se encuentra en su fase final. Intentaremos contar los inicios del trabajo cooperativo entre los universitarios –profesores y estudiantes- y los docentes ranqueles, la metodología de trabajo, la participación y gestión desarrollada a lo largo de los dos proyectos, los resultados en la formación de los estudiantes, y la evaluación de los resultados obtenidos

Enhanced CRAd Activity Using Enhancer Motifs Driven by a Nucleosome Positioning Sequence

Cancer development involves changes driven by the epigenetic machinery, including nucleosome positioning. Recently, the concept that adenoviral replication may be driven by tumor specific promoters (TSPs) gained support, and several conditionally replicative adenoviruses (CRAd) exhibited therapeutic efficacy in clinical trials. Here, we show for the first time that placing a nucleosome positioning sequence (NPS) upstream of a TSP combined with Wnt-responsive motifs (pART enhancer) enhanced the TSP transcriptional activity and increased the lytic activity of a CRAd. pART enhanced the transcriptional activity of the gastrointestinal cancer (GIC)-specific REG1A promoter (REG1A-pr); moreover, pART also increased the in vitro lytic activity of a CRAd whose replication was driven by REG1A-Pr. The pART enhancer effect in vitro and in vivo was strictly dependent on the presence of the NPS. Indeed, deletion of the NPS was strongly deleterious for the in vivo antitumor efficacy of the CRAd on orthotopically established pancreatic xenografts. pART also enhanced the specific activity ofmother heterologous promoters; moreover, the NPS was also able to enhance the responsiveness of hypoxia- and NFκ B-response elements. We conclude that NPS could be useful for gene therapy approaches in cancer as wellas other diseases.Fil: Bravo, Soraya. Universidad Andrés Bello; ChileFil: Núñez Aguilera, Felipe Javier. 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: Cruzat, Fernando. Universidad de Concepción; ChileFil: Cafferata, Eduardo Gustavo Alfredo. 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: De Ferrari, Giancarlo V. Universidad Andrés Bello; ChileFil: Montecino, Martín. Universidad Andrés Bello; ChileFil: Podhajcer, Osvaldo Luis. 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; Argentin

Spatio temporal population dynamics of the invasive diatom Didymosphenia geminate in central-southern Chilean rivers

We document the distribution of Didymosphenia geminata in central-southern Chilean rivers and identify the chemical and physical factors associated with its presence/absence (p/a). Repeated surveys in five successive years provided evidence that D. geminata could be nearing a biogeographic equilibrium in the region. D. geminata databases from extensive biological and environmental surveys in 187 rivers,within ten catchments, south of 38°S commenced in November 2010 and ran through May 2013. In addition, data from two other field surveys were used. The sites evenly distributed latitudinally were climatically characterized. The recent sampling program, following a published species distribution model, was designed to explore D. geminata distribution within thirteen catchments (34°S–48°S). An extensive river survey in 2014 (spring-summer) and in 2015 (autumn) included the p/a, and relative abundance of D. geminata cells in phytobenthos and in the drift. These p/a results showed that the probability of re-encountering D. geminata cells at sites where the species was previously found was significantly highwhile the probability of finding D. geminata cells at sites previously without the species was significantly low. This suggests that the distribution of D. geminata cells among suitable habitats was nearing completion. The relative abundance of D. geminata cells in the phytobenthos versus in the drift indicates seasonality with higher proportion of cells in the phytobenthos during the spring-summer than during the autumn. During the final surveys, principal component analysis of chemical and physical characteristics of rivers showed significant differences between rivers with and without D. geminata. Based on our observations of the distribution of D. geminata cells among rivers with suitable habitat conditions and the fluctuating rate of spread between rivers,we conclude that D. geminata is probably in the ending stage of its spatial demographic expansion in Chile surmounting the different barriers of the invasive process

Widespread loss of the silencing epigenetic mark H3K9me3 in astrocytes and neurons along with hippocampal-dependent cognitive impairment in C9orf72 BAC transgenic mice

Background: Hexanucleotide repeat expansions of the G4C2 motif in a non-coding region of the C9ORF72 gene are the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Tissues from C9ALS/FTD patients and from mouse models of ALS show RNA foci, dipeptide-repeat proteins, and notably, widespread alterations in the transcriptome. Epigenetic processes regulate gene expression without changing DNA sequences and therefore could account for the altered transcriptome profiles in C9ALS/FTD; here, we explore whether the critical repressive marks H3K9me2 and H3K9me3 are altered in a recently developed C9ALS/FTD BAC mouse model (C9BAC). Results: Chromocenters that constitute pericentric constitutive heterochromatin were visualized as DAPI- or Nucblue-dense foci in nuclei. Cultured C9BAC astrocytes exhibited a reduced staining signal for H3K9me3 (but not for H3K9me2) at chromocenters that was accompanied by a marked decline in the global nuclear level of this mark. Similar depletion of H3K9me3 at chromocenters was detected in astrocytes and neurons of the spinal cord, motor cortex, and hippocampus of C9BAC mice. The alterations of H3K9me3 in the hippocampus of C9BAC mice led us to identify previously undetected neuronal loss in CA1, CA3, and dentate gyrus, as well as hippocampal-dependent cognitive deficits. Conclusions: Our data indicate that a loss of the repressive mark H3K9me3 in astrocytes and neurons in the central nervous system of C9BAC mice represents a signature during neurodegeneration and memory deficit of C9ALS/FTD. © 2020 The Author(s).Indexación: Scopu

Novel approaches for immune reconstitution and adaptive immune modeling with human pluripotent stem cells

Pluripotent stem cells have the capacity to generate all cell lineages, and substantial progress has been made in realizing this potential. One fascinating but as yet unrealized possibility is the differentiation of pluripotent stem cells into thymic epithelial cells. The thymus is a primary lymphoid organ essential for naïve T-cell generation. T cells play an important role in adaptive immunity, and their loss or dysfunction underlies in a wide range of autoimmune and infectious diseases. T cells are generated and selected through interaction with thymic epithelial cells, the functionally essential element of thymus. The ability to generate functional thymic epithelial cells from pluripotent stem cells would have applications in modeling human immune responses in mice, in tissue transplantation, and in modulating autoimmune and infectious disease

T. brucei Infection Reduces B Lymphopoiesis in Bone Marrow and Truncates Compensatory Splenic Lymphopoiesis through Transitional B-Cell Apoptosis

African trypanosomes of the Trypanosoma brucei species are extracellular protozoan parasites that cause the deadly disease African trypanosomiasis in humans and contribute to the animal counterpart, Nagana. Trypanosome clearance from the bloodstream is mediated by antibodies specific for their Variant Surface Glycoprotein (VSG) coat antigens. However, T. brucei infection induces polyclonal B cell activation, B cell clonal exhaustion, sustained depletion of mature splenic Marginal Zone B (MZB) and Follicular B (FoB) cells, and destruction of the B-cell memory compartment. To determine how trypanosome infection compromises the humoral immune defense system we used a C57BL/6 T. brucei AnTat 1.1 mouse model and multicolor flow cytometry to document B cell development and maturation during infection. Our results show a more than 95% reduction in B cell precursor numbers from the CLP, pre-pro-B, pro-B, pre-B and immature B cell stages in the bone marrow. In the spleen, T. brucei induces extramedullary B lymphopoiesis as evidenced by significant increases in HSC-LMPP, CLP, pre-pro-B, pro-B and pre-B cell populations. However, final B cell maturation is abrogated by infection-induced apoptosis of transitional B cells of both the T1 and T2 populations which is not uniquely dependent on TNF-, Fas-, or prostaglandin-dependent death pathways. Results obtained from ex vivo co-cultures of living bloodstream form trypanosomes and splenocytes demonstrate that trypanosome surface coat-dependent contact with T1/2 B cells triggers their deletion. We conclude that infection-induced and possibly parasite-contact dependent deletion of transitional B cells prevents replenishment of mature B cell compartments during infection thus contributing to a loss of the host's capacity to sustain antibody responses against recurring parasitemic waves

Phytoplankton activity and standing crop in an impoundment of central Chile

Seasonal and spatial variations of the primary productivity were measured in Rapel reservoir, Chile in 1976 and 1977. Chlorophyll a, phytoplankton and several environmental factors were also measured. The results show that primary productivity began to increase in September, and remained at high levels until the following June near the dam. The same pattern, slightly displaced in time (October - May) was observed near the main river inflow areas. Standing crops were high at the beginning and at the end of those periods. The photosynthetic behavior of the phytoplankton was shown to be affected mainly by absolute light availability and the dimension of the illuminated water column. The phytoplankton composition and standing crop seems to be controlled by water temperature and flow. © 1982 IRL Press Ltd