11 research outputs found
The co-repressor mSin3A is a functional component of the REST-CoREST repressor complex
No full textThe repressor REST/NRSF restricts expression of a large set of genes to neurons by suppressing their expression in non-neural tissues. We find that REST repression involves two distinct repressor proteins. One of these, CoREST, interacts with the COOH-terminal repressor domain of REST (Andres, M. E., Burger, C., Peral-Rubio, M. J., Battaglioli, E., Anderson, M. E., Grimes, J., Dallmanm J., Ballas, N., and Mandel, G. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 9873-9878). Here we show that the co-repressor mSin3A also interacts with REST. The REST-mSin3A association involves the NH2-terminal repressor domain of REST and the paired amphipathic helix 2 domain of mSin3A. REST forms complexes with endogenous mSin3A in mammalian cells, and both mSin3A and CoREST interact with REST in intact mammalian cells. REST repression is blocked in yeast lacking Sin3 and rescued in its presence. In mammalian cells, repression by REST is reduced when binding to mSin3A is inhibited. In mouse embryos, the distribution of mSin3A and REST transcripts is largely coincident. The pattern of CoREST gene expression is more restricted, suggesting that mSin3A is required constitutively for REST repression, whereas CoREST is recruited for more specialized repressor functions
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Regulation of Neuronal Traits by a Novel Transcriptional Complex
No full textAbstractThe transcriptional repressor, REST, helps restrict neuronal traits to neurons by blocking their expression in nonneuronal cells. To examine the repercussions of REST expression in neurons, we generated a neuronal cell line that expresses REST conditionally. REST expression inhibited differentiation by nerve growth factor, suppressing both sodium current and neurite growth. A novel corepressor complex, CoREST/HDAC2, was shown to be required for REST repression. In the presence of REST, the CoREST/HDAC2 complex occupied the native Nav1.2 sodium channel gene in chromatin. In neuronal cells that lack REST and express sodium channels, the corepressor complex was not present on the gene. Collectively, these studies define a novel HDAC complex that is recruited by the C-terminal repressor domain of REST to actively repress genes essential to the neuronal phenotype
COVID-19 vaccine platforms: Delivering on a promise?
No full textThe emergence of the novel SARS-CoV-2 and COVID-19 has brought into sharp focus the need for a vaccine to prevent this disease. Vaccines have saved millions of lives since their introduction to the public over 200 years ago. The potential for vaccination reached new heights in the mid-20th century with the development of technologies that expanded the ability to create novel vaccines. Since then, there has been continued technological advancement in vaccine development. The resulting platforms provide the promise for solutions for many infectious diseases, including those that have been with us for decades as well as those just now emerging. Each vaccine platform represents a different technology with a unique set of advantages and challenges, especially when considering manufacturing. Therefore, it is essential to understand each platform as a separate product and process with its specific quality considerations. This review outlines the relevant platforms for developing a vaccine for SARS-CoV-2 and discusses the advantages and disadvantages of each
New high-pressure phase and equation of state of Ce2Zr2O8
No full textIn this paper we report a new high-pressure rhombohedral phase of Ce2Zr2O8 observed in high-pressure angle-dispersive x-ray diffraction and Raman spectroscopy studies up to nearly 12 GPa. The ambient-pressure cubic phase of Ce2Zr2O8 transforms to a rhombohedral structure beyond 5 GPa with a feeble distortion in the lattice. The pressure evolution of the unit-cell volume showed a change in compressibility above 5 GPa. The unit-cell parameters of the high-pressure rhombohedral phase at 12.1 GPa are a h = 14.6791(3) Å, c h = 17.9421(5) Å, and V = 3348.1(1) Å3. The structure relations between the parent cubic (P213) and rhombohedral (P32) phases were obtained via group-subgroup relations. All the Raman modes of the cubic phase showed linear evolution with pressure, with the hardest one at 197 cm¿1. Some Raman modes of the high-pressure phase have a non-linear evolution with pressure, and softening of one low-frequency mode with pressure is found. The compressibility, equation of state, and pressure coefficients of Raman modes of Ce2Zr2O8 are also reported. © 2012 American Institute of PhysicsErrandonea, D.; Kumar, R.; Achary, S.; Gomis Hilario, O.; Manjón Herrera, FJ.; Shukla, R.; Tyagi, A. (2012). New high-pressure phase and equation of state of Ce2Zr2O8. Journal of Applied Physics. 111:535191-5351910. doi:10.1063/1.3692807S5351915351910111Wuensch, B. (2000). Connection between oxygen-ion conductivity of pyrochlore fuel-cell materials and structural change with composition and temperature. Solid State Ionics, 129(1-4), 111-133. doi:10.1016/s0167-2738(99)00320-3Loong, C. K., Richardson, J. W., & Ozawa, M. (1995). Crystal Phases, Defects, and Dynamics of Adsorbed Hydroxyl Groups and Water in Pure and Lanthanide-Modified Zirconia: A Neutron-Scattering Study. Journal of Catalysis, 157(2), 636-644. doi:10.1006/jcat.1995.1329Ewing, R. C. (1999). Nuclear waste forms for actinides. 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Survey of peptide quantification methods and comparison of their reproducibility: A case study using oxytocin
No full textUSP's peptide reference standards content is typically determined using an HPLC assay against an external standard for which the purity was determined by a mass balance approach. To explore the use of other analytical methods, the USP Biologics Department conducted a multi-laboratory collaborative study. The study determined the inter-laboratory variability for peptide quantitation using the following methods: HPLC assay, quantitative nuclear magnetic resonance (qNMR) spectroscopy, or amino acid analysis (AAA). The three methods were compared with regard to their suitability for quantitation of the nonapeptide oxytocin. In this study, the HPLC assay method using the same peptide bulk material as the standard showed the lowest inter-lab variability. The coefficient of variation (%CV) was calculated without counting the uncertainty associated with the purity assignment of the standard with mass balance. The proton qNMR method is a direct measurement of the peptide against an internal standard, which is not difficult to perform under common laboratory conditions. Because of the simpler operation and shorter analytical time, qNMR as a primary method for peptide reference standard value assignment deserves further exploration
Unraveling Mechanisms of Transcriptional Repression: Novel Insights from Brinker
No full textTranscriptional repressors bind cis-regulatory elements of target genes in a sequence specific manner. To antagonize transcription, repressors primarily function by recruiting accessory proteins, co-repressors, which in turn largely function by modifying chromatin structure. Although a repressor could function by recruiting just a single co-repressor, many recruit more than one, with Brinker (Brk) from Drosophila recruiting the co-repressors, CtBP and Groucho (Gro), in addition to possessing a third repression domain, 3R. Previous studies indicated that Gro is sufficient for Brk to repress target genes in the wing imaginal disc, questioning why it should need to recruit CtBP, a ’short-range’ co-repressor compared to Gro that can function over longer distances. To resolve this I have used genomic engineering to generate a series of endogenous brk mutants that are unable to recruit Gro, CtBP and/or have the 3R domain deleted. Analysis of these mutants reveals that while the recruitment of Gro is necessary and is almost sufficient for Brk to make a morphologically wild-type fly, it is insufficient during oogenesis where Brk must utilize CtBP and 3R to pattern the egg-shell appropriately. Gro insufficiency during oogenesis can be explained by its downregulation in Brk-expressing cells through phosphorylation downstream of EGFR signaling, thus making it unavailable for Brk which must then resort to CtBP and/or 3R for repressive activity. The present study dissects the mechanism of activity of a transcription factor and its co-repressors and is the first to do so in multicellular eukaryotes in a physiologically relevant manner; additionally its findings provide a better understanding of why transcription factors in general may utilize more than one co-repressor
THE DETERMINANTS OF MIGRATION PROSPECT : SPATIAL ECONOMETRIC APPLICATION TO THE MOROCCAN DATA
No full textThe adepts of international migration have attempted to develop models that can be usedto examine the underlying factors that push the migrants to take the decision of leavingthe country of origin. The have tried to analyse the way in which the difference in thesocioeconomic determinants between spatial entities (countries, regions and provinces)may affect the decision of people to immigrate. This work has recommended first to verifyempirically the validation of the theory of migratory basins within one country and toidentify variables that clarify the intent of emigration. With respect to the Moroccancontext, this work presents a two-fold interest. On the methodological level, thiscontribution is, to our knowledge, the first application of the micro econometric spatialapproach on the data collected during our investigation. At the analytical level, thefindings of this work complement those of the few descriptive studies. Subsequently, theresults of this research should be used to reflect deeply upon the public policies related toemigration
WHO Expert Committee on Biological Standardization Introduction
No full textAfdeling Klinische Chemie en Laboratoriumgeneeskunde (AKCL
Expresión del receptor tipo toll 4 en células deciduales estromales humanas asociadas a complicaciones del embarazo
No full text107 p. CdLa Pre-eclampsia (PE) es una enfermedad multisistémica sin causa conocida que afecta entre el 5-7% de mujeres embarazadas, siendo una de las principales causas de morbilidad y mortalidad materno-fetal en todo el mundo. La asociación entre infección e inflamación es considerada la causa principal de complicaciones como la PE, con aumento de citoquinas favoreciendo la disfunción endotelial en placenta. Las células deciduales estromales (DSC) constituyen el principal componente celular de la decidua y protegen al feto frente a bacterias Gram negativas y señales de peligro que emanan de inflamación severa con o sin infección. A través de la expresión del receptor tipo Toll 4 (TLR4) las DSC regulan la respuesta inmune innata durante la infección. El objetivo fue analizar el comportamiento inmunomodulador in vitro de las DSC a partir de PE.
Materiales y métodos: estudio analítico de casos y controles. Las DSC fueron aisladas y cultivadas a partir de tejido placentario, de pacientes con parto vaginal sin complicaciones (control, n=6) y parto por cesárea con pre-eclampsia (PE, n=6). La caracterización fenotípica de las DSC se efectuó mediante análisis por citometría de flujo. La expresión del ARN mensajero del TLR4 se cuantificó por RT-PCR en tiempo real. Finalmente, la determinación de citoquinas se realizó por medio de ELISA a partir de sobrenadantes de cultivos celulares.
Resultados: la expresión de antígenos característicos fue similar en las DSC estudiadas (CD10, CD29, CD44, CD73 y CD90). La expresión génica del TLR4, no presentó diferencia significativa (p=0,136). Las DSC control secretaron mayor cantidad de IL-6 (media=3631,7pg/mL) en comparación con las de PE (760pg/mL). La secreción de IL-10 presento diferencia significativa (control=4pg/mL, PE11,2pg/mL). Conclusiones: las DSC control y PE no presentaron diferencias significativas en la expresión del TLR4, se sugiere un papel protector a nivel placentarioPre-eclampsia (PE) is a multisystemic disease with no known cause that affects 5-7% of pregnant women, being one of the main causes of maternal-fetal morbidity and mortality worldwide. The association between infection and inflammation is considered the main cause of complications such as PE, with increased cytokines favoring endothelial dysfunction in the placenta. Decidual stromal cells (DSC) constitute the main cellular component of the decidua and protect the fetus against Gram-negative bacteria and warning signs that emanate from severe inflammation with or without infection. Through the expression of Toll-like receptor 4 (TLR4), DSCs regulate the innate immune response during infection. The objective was to analyze the in vitro immunomodulatory behavior of DSCs from PE.
Materials and methods: analytical case and control study. The DSCs were isolated and cultured from placental tissue, from patients with uncomplicated vaginal delivery (control, n = 6) and cesarean delivery with pre-eclampsia (PE, n = 6). The phenotypic characterization of the DSC was carried out by means of flow cytometry analysis. The expression of the TLR4 messenger RNA was quantified by RT-PCR in real time. Finally, the determination of cytokines was performed by means of ELISA from supernatants of cell cultures.
Results: the expression of characteristic antigens was similar in the DSC studied (CD10, CD29, CD44, CD73 and CD90). Gene expression of TLR4 did not present a significant difference (p = 0.136). Control DSCs secreted a higher amount of IL-6 (mean = 3631.7pg / mL) compared to PE (760pg / mL). The secretion of IL-10 showed significant difference (control = 4pg / mL, PE11.2pg / mL). Conclusions: control DSC and PE did not present significant differences in the expression of the TLR4; a protective role at the placental level is suggested.TABLA DE CONTENIDO
INTRODUCCIÓN .......................................................................................... 14
1. PARTICIPANTES EN EL DESARROLLO DEL EMBARAZO .................... 15
1.1 CÉLULAS DECIDUALES ESTROMALES ........................................... 15
1.1.1 Fenotipo antigénico de las DSC .................................................... 16
1.2 INTERFASE MATERNO-FETAL ......................................................... 19
1.3 COMPLICACIONES DEL EMBARAZO ............................................... 22
1.4 PRE-ECLAMPSIA ................................................................................ 24
1.5 ESTRUCTURA Y SEÑALIZACIÓN DEL RECEPTOR TIPO TOLL – 4 31
1.5.1. Expresión de TLR4 en placentas asociadas a patologías del embarazo ............................................................................................... 35
1.6 CITOQUINAS EN EL EMBARAZO ...................................................... 38
1.7 INFECCIÓNES DURANTE EL EMBARAZO ....................................... 41
1.7.1 Enfermedad periodontal y complicaciones del embarazo ............ 44
1.7.2 Infección urinaria y pre-eclampsia ................................................ 47
2. OBJETIVOS .............................................................................................. 50
2.1 OBJETIVO GENERAL ......................................................................... 50
2.2 OBJETIVOS ESPECÍFICOS ............................................................... 50
3. MATERIALES Y MÉTODOS ..................................................................... 51
3.1 DEFINICIÓN DE SUJETOS DE ESTUDIO .......................................... 51
3.1.1 Aspectos éticos del proyecto ......................................................... 51
3.1.2 Criterios de inclusión ..................................................................... 52
3.1.3 Criterios de exclusión ................................................................... 52
3.1.4 Obtención de las muestras ............................................................ 52
3.2 CULTIVO CELULAR ............................................................................ 53
3.2.1 Obtención, purificación y cultivo de las DSC ................................. 53
3.2.2 Recolección de sobrenadantes ..................................................... 54
3.3 CITOMETRÍA DE FLUJO .................................................................... 54
3.3.1 Análisis del fenotipo antigénico de las DSC .................................. 54
3.4 ENSAYO RT-PCR EN TIEMPO REAL ................................................ 55
3.4.1 Preparación del cultivo de DSC ..................................................... 55
3.4.2 Extracción de ARN total ................................................................ 55
3.4.3 RT-PCR en tiempo real ................................................................. 56
3.5 PRUEBA DE ELISA ............................................................................. 57
3.5.1 Detección de citoquinas ................................................................ 57
3.6 ANÁLISIS ESTADÍSTICO .................................................................... 57
4. RESULTADOS .......................................................................................... 58
4.1 CONDICIÓN CLÍNICA DE LAS PARTICIPANTES .............................. 58
4.2 CARACTERIZACIÓN FENOTIPICA DE LAS DSC .............................. 59
4.2.1 Aislamiento de DSC obtenidas de placentas ................................. 59
4.2.2. Fenotipo antigénico de DSC cultivadas ........................................ 60
4.3 EXPRESIÓN GÉNICA DEL TLR4 POR LAS DSC .............................. 62
4.3.1 Extracción e integridad del ARN .................................................... 62
4.3.2 Eficiencia de primers ..................................................................... 63
4.3.3 Expresión del TLR4 ....................................................................... 64
4.4 SECRECIÓN DE CITOQUINAS .......................................................... 70
4.4.1 Secreción de IL-6 por las células DSC en cultivo, control y PE ..... 70
4.4.2 Secreción de IL-10 por las células DSC en cultivo, control y PE ... 71
5. DISCUSIÓN .............................................................................................. 72
6. CONCLUSIONES ..................................................................................... 85
7. REFERENCIAS ........................................................................................ 86Ej. 1MaestríaMagister en Investigación en enfermedades Infecciosa
