260 research outputs found
Interplay of multivalency and optical properties of quantum dots: implications for sensing and actuation in living cells.
Educação, conflito e convivĂȘncia democrĂĄtica
Após uma caracterização sucinta da actual condição pós-moderna, serão desenvolvidas
algumas questÔes que, pela sua relevùncia actual no campo da educação,
merecem ser revisitadas de modo crĂtico, nomeadamente, e num primeiro momento,
a escola, o conflito e a convivĂȘncia. Num segundo momento, apontar-se-ĂŁo algumas
caracterĂsticas da escola que fazem dela uma organização com alguma perversidade,
hipocrisia e irracionalidade. O Ășltimo aspecto a ser tratado irĂĄ compreender
a escola como organização comunicacional ou organização convivencial, onde os
conceitos de disciplina, violĂȘncia, conflito e convivĂȘncia assumem um sentido mais
profundamente democrĂĄtico.After a succinct characterization of the current postmodern condition, some
questions might be developed, because of its current relevance in the education
field, they must to be reviewed in a critical way, namely, and firstly, the school, the
conflict and the conviviality. Secondly, this article underlines some schoolâs
characteristics that turns it into an organization with a little perversity, hypocrisy
and irrationality. The last aspect to be treated is the understanding of the school as
a communicative or convivial organization, in which the concepts of discipline,
violence, conflict and conviviality assume a more democratic orientation.Enseguida a una caracterizaciĂłn sucinta de la actual condiciĂłn pos-moderna,
serĂĄn desarrolladas algunas cuestiones que, por su relevancia actual en el campo
de la educaciĂłn, merecen ser visitadas de modo crĂtico, nombradamente, y en
primero momento, la escuela, el conflicto y la convivencia. En segundo momento,
se apuntarĂĄn algunas caracterĂsticas de la escuela que la hacen una organizaciĂłn
con alguna perversidad, hipocresĂa e irracionalidad. El Ășltimo aspecto a ser tratado irĂĄ comprender la escuela como organizaciĂłn de comunicaciĂłn u
organizaciĂłn de convivencia, donde los conceptos de disciplina, violencia,
conflicto e convivencia asumen un sentido mĂĄs profundamente democrĂĄtico.(undefined
MicroRNAs expression, chromosomal alterations and immunoglobulin variable Heavy chain hypermutations in Mantle Cell Lymphomas
The contribution of microRNAs (miR) to the pathogenesis of mantle cell lymphoma (MCL) is not well known.We investigated
the expression of 86 mature miRs mapped to frequently altered genomic regions in MCL in CD5+/CD5 normal B cells, reactive
lymph nodes, and purified tumor cells of 17 leukemic MCL, 12 nodal MCL, and 8MCL cell lines. Genomic alterations of the
tumors were studied by single nucleotide polymorphism arrays
and comparative genomic hybridization. Leukemic and nodal
tumors showed a high number of differentially expressed miRs
compared with purified normal B cells, but only some of them
were commonly deregulated in both tumor types. An unsupervised
analysis of miR expression profile in purified leukemic
MCL cells revealed two clusters of tumors characterized by
different mutational status of the immunoglobulin genes,
proliferation signature, and number of genomic alterations.
The expression of most miRs was not related to copy number
changes in their respective chromosomal loci. Only the levels
of miRs included in the miR-17-92 cluster were significantly
related to genetic alterations at 13q31. Moreover, overexpression
of miR-17-5p/miR-20a from this cluster was associated
with high MYC mRNA levels in tumors with a more aggressive
behavior. In conclusion, the miR expression pattern of MCL is
deregulated in comparison with normal lymphoid cells and
distinguishes two subgroups of tumors with different biological
features.Postprint (updated version
Identification of Methylated Genes Associated with Aggressive Clinicopathological Features in Mantle Cell Lymphoma
Background: Mantle cell lymphoma (MCL) is genetically characterized by the t(11;14)(q13;q32) translocation and a high number of secondary chromosomal alterations. The contribution of DNA methylation to MCL lymphomagenesis is not well known. We sought to identify epigenetically silenced genes in these tumours that might have clinical relevance. Methodology/Principal Findings: To identify potential methylated genes in MCL we initially investigated seven MCL cell lines treated with epigenetic drugs and gene expression microarray profiling. The methylation status of selected candidate genes was validated by a quantitative assay and subsequently analyzed in a series of primary MCL (n=38). After pharmacological reversion we identified 252 potentially methylated genes. The methylation analysis of a subset of these genes (n=25) in the MCL cell lines and normal B lymphocytes confirmed that 80% of them were methylated in the cell lines but not in normal lymphocytes. The subsequent analysis in primary MCL identified five genes (SOX9,HOXA9,AHR,NR2F2 ,and ROBO1) frequently methylated in these tumours. The gene methylation events tended to occur in the same primary neoplasms and correlated with higher proliferation, increased number of chromosomal abnormalities, and shorter survival of the patients. Conclusions: We have identified a set of genes whose methylation degree and gene expression levels correlate with aggressive clinicopathological features of MCL. Our findings also suggest that a subset of MCL might show a CpG island methylator phenotype (CIMP) that may influence the behaviour of the tumours
The GTPase RalA Regulates Different Steps of the Secretory Process in Pancreatic ÎČ-Cells
BACKGROUND: RalA and RalB are multifuntional GTPases involved in a variety of cellular processes including proliferation, oncogenic transformation and membrane trafficking. Here we investigated the mechanisms leading to activation of Ral proteins in pancreatic beta-cells and analyzed the impact on different steps of the insulin-secretory process. METHODOLOGY/PRINCIPAL FINDINGS: We found that RalA is the predominant isoform expressed in pancreatic islets and insulin-secreting cell lines. Silencing of this GTPase in INS-1E cells by RNA interference led to a decrease in secretagogue-induced insulin release. Real-time measurements by fluorescence resonance energy transfer revealed that RalA activation in response to secretagogues occurs within 3-5 min and reaches a plateau after 10-15 min. The activation of the GTPase is triggered by increases in intracellular Ca2+ and cAMP and is prevented by the L-type voltage-gated Ca2+ channel blocker Nifedipine and by the protein kinase A inhibitor H89. Defective insulin release in cells lacking RalA is associated with a decrease in the secretory granules docked at the plasma membrane detected by Total Internal Reflection Fluorescence microscopy and with a strong impairment in Phospholipase D1 activation in response to secretagogues. RalA was found to be activated by RalGDS and to be severely hampered upon silencing of this GDP/GTP exchange factor. Accordingly, INS-1E cells lacking RalGDS displayed a reduction in hormone secretion induced by secretagogues and in the number of insulin-containing granules docked at the plasma membrane. CONCLUSIONS/SIGNIFICANCE: Taken together, our data indicate that RalA activation elicited by the exchange factor RalGDS in response to a rise in intracellular Ca2+ and cAMP controls hormone release from pancreatic beta-cell by coordinating the execution of different events in the secretory pathway
Latin American anaphylaxis registry
Background: Recent data about clinical features, triggers and management of anaphylaxis in Latin America is lacking. Objective: To provide updated and extended data on anaphylaxis in this region. Method: An online questionnaire was used, with 67 allergy units involved from 12 Latin-American countries and Spain. Among data recorded, demographic information, clinical features, severity, triggering agents, and treatment were received. Results: Eight hundred and seventeen anaphylactic reactions were recorded. No difference in severity, regardless of pre-existing allergy or asthma history was found. Drug induced anaphylaxis (DIA) was most frequent (40.6%), followed by food induced anaphylaxis (FIA) (32.9%) and venom induced anaphylaxis (VIA) (12%). FIA and VIA were more common in children-adolescents. Non-steroidal anti-inflammatory drugs (NSAIDs) and beta-lactam antibiotics (BLA) were the most frequent drugs involved. Milk (61.1% of FIA) and egg (15.4% of FIA) in children, and shellfish (25.5% of FIA), fresh fruits (14.2% of FIA), and fish (11.3% of FIA) in adults were the most common FIA triggers. Fire ants were the most frequent insect triggers, and they induced more severe reactions than triggers of FIA and DIA (p < 0.0001). Epinephrine was used in 43.8% of anaphylaxis episodes. After Emergency Department treatment, epinephrine was prescribed to 13% of patients. Conclusions: Drugs (NSAIDs and BLA), foods (milk and egg in children and shellfish, fruits and fish in adults) and fire ants were the most common inducers of anaphylaxis. Epinephrine was used in less than half of the episodes emphasizing the urgent need to improve dissemination and implementation of anaphylaxis guidelines.RevisiĂłn por pare
Temporal resolution of proteinâprotein interactions in the live-cell plasma membrane
We have recently devised a method to quantify interactions between a membrane protein (âbaitâ) and a fluorophore-labeled protein (âpreyâ) directly in the live-cell plasma membrane (Schwarzenbacher et al. Nature Methods 5:1053â1060 2008). The idea is to seed cells on surfaces containing micro-patterned antibodies against the exoplasmic domain of the bait, and monitor the co-patterning of the fluorescent prey via fluorescence microscopy. Here, we characterized the time course of bait and prey micropattern formation upon seeding the cells onto the micro-biochip. Patterns were formed immediately after contact of the cells with the surface. Cells were able to migrate over the chip surface without affecting the micropattern contrast, which remained constant over hours. On single cells, bait contrast may be subject to fluctuations, indicating that the bait can be released from and recaptured on the micropatterns. We conclude that interaction studies can be performed at any time-point ranging from 5Â min to several hours post seeding. Monitoring interactions with time opens up the possibility for new assays, which are briefly sketched in the discussion section
New Algorithm to Determine True Colocalization in Combination with Image Restoration and Time-Lapse Confocal Microscopy to Map Kinases in Mitochondria
The subcellular localization and physiological functions of biomolecules are closely related and thus it is crucial to precisely determine the distribution of different molecules inside the intracellular structures. This is frequently accomplished by fluorescence microscopy with well-characterized markers and posterior evaluation of the signal colocalization. Rigorous study of colocalization requires statistical analysis of the data, albeit yet no single technique has been established as a standard method. Indeed, the few methods currently available are only accurate in images with particular characteristics. Here, we introduce a new algorithm to automatically obtain the true colocalization between images that is suitable for a wide variety of biological situations. To proceed, the algorithm contemplates the individual contribution of each pixel's fluorescence intensity in a pair of images to the overall PearsoĆs correlation and Manders' overlap coefficients. The accuracy and reliability of the algorithm was validated on both simulated and real images that reflected the characteristics of a range of biological samples. We used this algorithm in combination with image restoration by deconvolution and time-lapse confocal microscopy to address the localization of MEK1 in the mitochondria of different cell lines. Appraising the previously described behavior of Akt1 corroborated the reliability of the combined use of these techniques. Together, the present work provides a novel statistical approach to accurately and reliably determine the colocalization in a variety of biological images
Epigenetic Activation of SOX11 in Lymphoid Neoplasms by Histone Modifications
Recent studies have shown aberrant expression of SOX11 in various types of aggressive B-cell neoplasms. To elucidate the molecular mechanisms leading to such deregulation, we performed a comprehensive SOX11 gene expression and epigenetic study in stem cells, normal hematopoietic cells and different lymphoid neoplasms. We observed that SOX11 expression is associated with unmethylated DNA and presence of activating histone marks (H3K9/14Ac and H3K4me3) in embryonic stem cells and some aggressive B-cell neoplasms. In contrast, adult stem cells, normal hematopoietic cells and other lymphoid neoplasms do not express SOX11. Such repression was associated with silencing histone marks H3K9me2 and H3K27me3. The SOX11 promoter of non-malignant cells was consistently unmethylated whereas lymphoid neoplasms with silenced SOX11 tended to acquire DNA hypermethylation. SOX11 silencing in cell lines was reversed by the histone deacetylase inhibitor SAHA but not by the DNA methyltransferase inhibitor AZA. These data indicate that, although DNA hypermethylation of SOX11 is frequent in lymphoid neoplasms, it seems to be functionally inert, as SOX11 is already silenced in the hematopoietic system. In contrast, the pathogenic role of SOX11 is associated with its de novo expression in some aggressive lymphoid malignancies, which is mediated by a shift from inactivating to activating histone modifications
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