Políticas públicas para erradicação do trabalho escravo contemporâneo no Brasil: um estudo sobre a dinâmica das relações entre os atores governamentais e não-governamentais

O estudo tem como objetivo identificar na evolução das políticas públicas para erradicação do trabalho escravo os diferentes atores e a dinâmica das relações entre eles. A ocorrência da escravidão contemporânea pôde se dar a partir da contribuição de alguns fatores estruturais e conjunturais, tais como o processo de aprofundamento do capitalismo e de modernização conservadora no país e especificamente na agricultura e relações políticas, sociais e históricas que perpetuam a enorme concentração fundiária brasileira. Além disso, algumas relações pessoais, sociais e políticas de intermediação de interesses entre Estado e sociedade, tais como clientelismo e patronagem e redes de políticas, de modo geral e de forma mais específica nas políticas agrárias, também interferem no desenvolvimento dos processos de políticas públicas e dentre elas nas políticas de combate ao trabalho escravo. Desse modo, a dissertação tem como problema a investigação da dinâmica das relações entre atores governamentais e nãogovernamentais na formulação e implantação das políticas públicas de erradicação ao trabalho escravo no Brasil. Para tanto, o estudo foi realizado por meio de pesquisa bibliográfica, documental e de campo, tendo entrevistado os seguintes atores políticos: MTE, MPT, OIT, CPT, ONG Repórter Brasil, GPTEC e OAB. Os dados foram analisados pelo método de análise de conteúdo, sob um viés qualitativo. Os resultados da pesquisa permitiram identificar a formação de múltiplas redes entre os atores governamentais e não-governamentais envolvidos nesta questão, demonstrando certa divisão entre as redes que atuam lutando pelo combate ao trabalho escravo e outras que se posicionam como uma certa resistência a esse combate, devido a interesses econômicos e políticos, revelando, assim, um jogo de forças que ora apresenta avanços e conquistas, ora mostra retrocessos ou estagnação na luta contra a escravidão contemporânea brasileira.

Avaliação da eficiência de co-transformação de soja (Glycine max (L.) Merrill) via bombardeamento de partículas utilizando um gene de seleção e um gene de interesse em plamídeos diferentes

Visando aumentar a resistência a moléstias fúngicas, o presente trabalho teve como objetivo introduzir um gene (chit1) que codifica uma quitinase do fungo Metarhizium anisopliae em cultivares de soja [Glycine max (L.) Merrill]. A co-transformação foi a estratégia escolhida, visando a obtenção de plantas livres de transgenes marcadores na progênie das plantas transformadas. A co-transformação foi realizada via biolística, tendo como tecido-alvo conjuntos de embriões somáticos globulares das cultivares MG/BR46 Conquista e IAS-5. O plasmídeo pGusHyg, que contém o gene repórter gusA e o gene marcador hpt, foi bombardeado concomitantemente com o plasmídeo pMOG463chit1, que porta o gene chit1. Os conjuntos de embriões bombardeados foram transferidos para meio seletivo contendo higromicina, visando a obtenção de material estavelmente transformado. Os conjuntos embriogênicos higromicina-resistentes foram transferidos seqüencialmente para meios de proliferação D-20 (sem higromicina), maturação e regeneração. No total, foram obtidos 387 e 380 embriões histodiferenciados das cultivares MG/BR46 Conquista e IAS-5, respectivamente. Plantas transgênicas adultas e férteis foram regeneradas. Para avaliar a eficiência da estratégia de cotransformação, foram realizadas análises moleculares de embriões histodiferenciados e de plantas regeneradas. Os resultados obtidos neste trabalho permitiram o cálculo da taxa de co-transformação de 44% para os embriões histodiferenciados da cultivar MG/BR46 Conquista e de 50% para plantas de IAS-5. Não existem, até o momento, relatos de trabalhos em soja utilizando embriões somáticos globulares em proliferação como alvo para estudos de co-transformação.

Profissão: repórter esportivo - das narrativas da nação ao ofício do jornalista contemporâneo

Essa dissertação tem por objetivo investigar de que maneira o repórter esportivo contemporâneo se relaciona com o futebol e como essa relação interfere no seu ofício e na construção do texto jornalístico. Mais especificamente, busca identificar as práticas adotadas e as estratégias narrativas utilizadas na construção de seus textos. O estudo dos elementos presentes na prática jornalística tem como perspectiva que a mídia participou ativamente da construção do imaginário futebolístico nacional e é na contemporaneidade a grande mediadora entre o público e os eventos esportivos. É sob este prisma que se pretende apurar como se dá a relação com o trabalho do profissional de imprensa que se identifica com seu objeto, analisar qual o lugar da emoção na composição da notícia esportiva e verificar que valores imbuem esse profissional na construção do texto jornalístico. A análise tem como corpus entrevistas de história oral temática com seis jornalistas que atuam ou atuaram como repórteres esportivos em jornais diários da cidade do Rio de Janeiro, todos com expressiva trajetória no meio esportivo, o que lhes confere credibilidade e autoridade públicas para protagonizar a constituição da memória da imprensa esportiva brasileira. O trabalho é constituído por três etapas: 1) a contextualização – quando se relaciona futebol e formação social brasileira, imprensa e jornalismo esportivo -; 2) o estudo do jornalismo como ofício a partir das teorias da Sociologia das Profissões e das estratégias narrativas utilizadas no texto noticioso esportivo nos jornais; e 3) a análise qualitativa das entrevistas temáticas.

Business diplomacy: the compass rose of foreign markets

With transnational corporations (TNCs) around the world today numbering over 60,000 and more than 800,000 affiliates working abroad, it is easy to understand how modern day international business could have transformed into a major global player serving at the axis of politics, social and environmental responsibility. Additionally, with accountability to a large variety of both public and private stakeholders, all exerting significant power and influence, today’s global corporate structure is reinventing modern international relations, and in some cases, dominating it. (Muldoon 2005) This transformative nature of globalization today can also serve as a source of friction among this growing chorus of players and is bringing irreversible change to these relationships and how they impact and influence business around the world. (Muldoon 2005) From the largest to the smallest international corporation seeking to expand into new international markets, the challenges that come with corporate ambition can mean the difference between success and failure and they find a home at the intersection of international relations, diplomacy and economics. To successfully navigate these challenges, especially in emerging economies, a company must now factor in more than just the “bottom line” and address complex issues that include human rights differences, environmental regulations, labor rights and values of each country. (Henisz, 2014) Combined with modern-day mobility achieved through technology and the Internet, corporations today have a great capacity to reach targeted audiences and establish a presence, but it is this same technology that also allows for immediate response to any corporate action. This constant, 24-hour news cycle, where everyone is made to be a real-time reporter through social media, has created a situation that demonstrably necessitates the ability to not only 3 respond immediately, but also to have real-time understanding of the challenges faced by a corporation as it looks toward global expansion. International Business Diplomacy, or simply Business Diplomacy as it will be referred to in this paper, combines all of these nuanced factors into a relatively new discipline that offers companies looking to expand into new markets, guidelines and directives so that they can more strategically map corporate direction, limit risk and achieve their objectives. This paper will examine the history of diplomacy and how the concept of statecraft became intertwined with the increasing globalization of business. Following a scholarly examination of how modern Business Diplomacy came into being, and the unique challenges that come with its application, particularly the liabilities needed to be overcome, this paper will apply the concept to the Brazilian aerospace manufacturer Embraer, tracking its strategic emergence from a small, regionally focused aircraft producer to global leader in the regional and executive jet market platforms. It will then examine Embraer’s entrance into the Chinese market, where the company suffered from several missteps and eventually had to refocus its business model from commercial to executive jets. Finally, as globalization continues to “emancipate international business from its institutional and social constraints,” (Muldoon 2005) this paper will address how the relatively new and emerging discipline of Business Diplomacy is continuing to mature and grow in stature and influence through the proposition of a new challenge or “liability” that corporations must also overcome as they expand into new markets. Through the analysis of Embraer in China, this paper will introduce the Liability of Governance to the lexicon of Business Diplomacy and propose specific steps that a company can undertake to avoid it.

Functionalization of dendrimers for improved gene delivery to mesenchymal stem cell

Disease, injury, and age problems compromise human quality of life and continuously motivate the search for new and more efficacious therapeutic approaches. The field of Tissue Regeneration and Engineering has greatly evolved over the last years, mainly due to the combination of the important advances verified in Biomaterials Science and Engineering with those of Cell and Molecular Biology. In particular, a new and promising area arose – Nanomedicine – that takes advantage of the extremely small size and especial chemical and physical properties of Nanomaterials, offering powerful tools for health improvement. Research on Stem Cells, the self-renewing progenitors of body tissues, is also challenging to the medical and scientific communities, being expectable the appearance of new and exciting stem cell-based therapies in the next years. The control of cell behavior (namely, of cell proliferation and differentiation) is of key importance in devising strategies for Tissue Regeneration and Engineering. Cytokines, growth factors, transcription factors and other signaling molecules, most of them proteins, have been identified and found to regulate and support tissue development and regeneration. However, the application of these molecules in long-term regenerative processes requires their continuous presence at high concentrations as they usually present short half-lives at physiological conditions and may be rapidly cleared from the body. Alternatively, genes encoding such proteins can be introduced inside cells and be expressed using cell’s machinery, allowing an extended and more sustained production of the protein of interest (gene therapy). Genetic engineering of stem cells is particularly attractive because of their self-renewal capability and differentiation potential. For Tissue Regeneration and Engineering purposes, the patient’s own stem cells can be genetically engineered in vitro and, after, introduced in the body (with or without a scaffold) where they will not only modulate the behavior of native cells (stem cell-mediated gene therapy), but also directly participate in tissue repair. Cells can be genetically engineered using viral and non-viral systems. Viruses, as a result of millions of years of evolution, are very effective for the delivery of genes in several types of cells, including cells from primary sources. However, the risks associated with their use (like infection and immunogenic reactions) are driving the search for non-viral systems that will efficiently deliver genetic material into cells. Among them, chemical methods that are promising and being investigated use cationic molecules as carriers for DNA. In this case, gene delivery and gene expression level remain relatively low when primary cells are used. The main goal of this thesis was to develop and assess the in vitro potential of polyamidoamine (PAMAM) dendrimers based carriers to deliver genes to mesenchymal stem cells (MSCs). PAMAM dendrimers are monodispersive, hyperbranched and nanospherical molecules presenting unique characteristics that make them very attractive vehicles for both drug and gene delivery. Although they have been explored for gene delivery in a wide range of cell lines, the interaction and the usefulness of these molecules in the delivery of genes to MSCs remains a field to be explored. Adult MSCs were chosen for the studies due to their potential biomedical applications (they are considered multipotent cells) and because they present several advantages over embryonic stem cells, such as easy accessibility and the inexistence of ethical restrictions to their use. This thesis is divided in 5 interconnected chapters. Chapter I provides an overview of the current literature concerning the various non-viral systems investigated for gene delivery in MSCs. Attention is devoted to physical methods, as well as to chemical methods that make use of polymers (natural and synthetic), liposomes, and inorganic nanoparticles as gene delivery vectors. Also, it summarizes the current applications of genetically engineered mesenchymal stem cells using non-viral systems in regenerative medicine, with special focus on bone tissue regeneration. In Chapter II, the potential of native PAMAM dendrimers with amine termini to transfect MSCs is evaluated. The level of transfection achieved with the dendrimers is, in a first step, studied using a plasmid DNA (pDNA) encoding for the β-galactosidase reporter gene. The effect of dendrimer’s generation, cell passage number, and N:P ratio (where N= number of primary amines in the dendrimer; P= number of phosphate groups in the pDNA backbone) on the level of transfection is evaluated, being the values always very low. In a second step, a pDNA encoding for bone morphogenetic protein-2, a protein that is known for its role in MSCs proliferation and differentiation, is used. The BMP-2 content produced by transfected cells is evaluated by an ELISA assay and its effect on the osteogenic markers is analyzed through several classical assays including alkaline phosphatase activity (an early marker of osteogenesis), osteocalcin production, calcium deposition and mineralized nodules formation (late osteogenesis markers). Results show that a low transfection level is enough to induce in vitro osteogenic differentiation in MSCs. Next, from Chapter III to Chapter V, studies are shown where several strategies are adopted to change the interaction of PAMAM dendrimers with MSCs cell membrane and, as a consequence, to enhance the levels of gene delivery. In Chapter III, generations 5 and 6 of PAMAM dendrimers are surface functionalized with arginine-glycine-aspartic acid (RGD) containing peptides – experiments with dendrimers conjugated to 4, 8 and 16 RGD units were performed. The underlying concept is that by including the RGD integrin-binding motif in the design of the vectors and by forming RGD clusters, the level of transfection will increase as MSCs highly express integrins at their surface. Results show that cellular uptake of functionalized dendrimers and gene expression is enhanced in comparison with the native dendrimers. Furthermore, gene expression is dependent on both the electrostatic interaction established between the dendrimer moiety and the cell surface and the nanocluster RGD density. In Chapter IV, a new family of gene delivery vectors is synthesized consisting of a PAMAM dendrimer (generation 5) core randomly linked at the periphery to alkyl hydrophobic chains that vary in length and number. Herein, the idea is to take advantage of both the cationic nature of the dendrimer and the capacity of lipids to interact with biological membranes. These new vectors show a remarkable capacity for internalizing pDNA, being this effect positively correlated with the –CH2– content present in the hydrophobic corona. Gene expression is also greatly enhanced using the new vectors but, in this case, the higher efficiency is shown by the vectors containing the smallest hydrophobic chains. Finally, chapter V reports the synthesis, characterization and evaluation of novel gene delivery vectors based on PAMAM dendrimers (generation 5) conjugated to peptides with high affinity for MSCs membrane binding - for comparison, experiments are also done with a peptide with low affinity binding properties. These systems present low cytotoxicity and transfection efficiencies superior to those of native dendrimers and partially degraded dendrimers (Superfect®, a commercial product). Furthermore, with this biomimetic approach, the process of gene delivery is shown to be cell surface receptor-mediated. Overall, results show the potential of PAMAM dendrimers to be used, as such or modified, in Tissue Regeneration and Engineering. To our knowledge, this is the first time that PAMAM dendrimers are studied as gene delivery vehicles in this context and using, as target, a cell type with clinical relevancy. It is shown that the cationic nature of PAMAM dendrimers with amine termini can be synergistically combined with surface engineering approaches, which will ultimately result in suitable interactions with the cytoplasmic membrane and enhanced pDNA cellular entry and gene expression. Nevertheless, the quantity of pDNA detected inside cell nucleus is always very small when compared with the bigger amount reaching cytoplasm (accumulation of pDNA is evident in the perinuclear region), suggesting that the main barrier to transfection is the nuclear membrane. Future work can then be envisaged based on the versatility of these systems as biomedical molecular materials, such as the conjugation of PAMAM dendrimers to molecules able to bind nuclear membrane receptors and to promote nuclear translocation.

Compreensão do típico ideal de técnicos de enfermagem acerca da sistematização da assistência de enfermagem

This study aimed to understand the typical ideal of the nursing technician about the systematization of nursing care in the light of the theoretical framework of Alfred Schutz. It is a comprehensive phenomenological research, using the theoretical framework of Alfred Schutz. For the unveiling of the phenomenon (the typical ideal of the nursing technician about the systematization of nursing care), the search process was configured from the proposed guiding principles for a research methodology based on the work itself of Schutz held by Zeferino (2010) in his PhD. For data collection, we used the focus group technique, counting on the collaboration of thirteen practical nurses working in a university hospital in Rio Grande do Norte, who responded positively to the inclusion criteria: working in the study hospital, performing care direct to patients. Forty-four subjects showed interest in participating, being held a draw for selection of the research sample, consisting of 14 professionals, one of whom did not attend the gathering of data collection. The focus group, entitled "What I think about the systematization of nursing care", took place on February 15th, 2013, totaling 101 minutes. It was performed according to the Experiential Education Humanescent using building posters as projective technique, from the key question: "What is the systematization of nursing care for you?". In order to understand some of the biographical situation of the participants, a questionnaire was administered to study participants. From the agreement of the subjects, the focus group was recorded and photographed with the cooperation of one reporter and two other employees. We used Microsoft Word 2010 to perform the transcript of the meeting and Microsoft Excel 2010 for synthesizing the results via a spreadsheet. The study followed the ethical and legal principles that govern scientific research on humans, recommended in Resolution nº 196/96, it was approved by Opinion Embodied Ethics Committee in Research of UFRN (Federal University of Rio Grande do Norte), nº 98 424, of August 31th, 2012, CAAE No. 05906912.0.0000.5537. The analysis of the nursing staff speeches, along with the contemplation of their posters and their written descriptions, allowed from the guiding principles of Zeferino (2010), in light of the reference of Alfred Schutz, unveiling the typical ideal of nursing technicians about the systematization of nursing care, passing four themes: typing of the concept of systematization of nursing care; benefits, which resulted in the reasons to believe in the positivity of this working tool; experienced problems, revealing the world of everyday life of nursing professionals, and possibilities for improvement. It was concluded that the nursing technicians are unaware of the systematization of nursing care. However, they typify a very positive perception about the same, especially with regard to improvements that may foster care

Cellular and molecular characterization of an embryonic cell line (BME26) from the tick Rhipicephalus (Boophilus) microplus

The cellular and molecular characteristics of a cell line (BME26) derived from embryos of the cattle tick Rhipicephalus (Boophilus) microplus were studied. The cells contained glycogen inclusions, numerous mitochondria, and vesicles with heterogeneous electron densities dispersed throughout the cytoplasm. Vesicles contained lipids and sequestered palladium meso-porphyrin (Pd-mP) and rhodamine-hemoglobin, suggesting their involvement in the autophagic and endocytic pathways. The cells phagocytosed yeast and expressed genes encoding the antimicrobial peptides (microplusin and defensin). A cDNA library was made and 898 unique mRNA sequences were obtained. Among them, 556 sequences were not significantly similar to any sequence found in public databases. Annotation using Gene Ontology revealed transcripts related to several different functional classes. We identified transcripts involved in immune response such as ferritin, serine proteases, protease inhibitors,. antimicrobial peptides, heat shock protein, glutathione S-transferase, peroxidase, and NADPH oxidase. BME26 cells transfected with a plasmid carrying a red fluorescent protein reporter gene (DsRed2) transiently expressed DsRed2 for up to 5 weeks. We conclude that BME26 can be used to experimentally analyze diverse biological processes that occur in R. (B.) microplus such as the innate immune response to tick-borne pathogens. (C) 2008 Elsevier Ltd. All rights reserved.

A heterogeneidade discursiva em revistas de divulgação científica

This research aiming at show an interpretative description about the form and function of Scientific Publication (SP) discursive genre, in two magazines of national circulation. We analyzed subjects published from 2004 to 2006, in the magazines Revista do Professor and Revista Nova Escola. We see at SP subjects reported discourses, into its two main presentation forms of other voices: direct discourse and indirect discourse. We have established some aims, first, we analyzing different forms to mark the discursive heterogeneity, by the reason the writer conceptualize an image of his/her interlocutor. The second one, we intend to look at the differences between marked heterogeneity according to the writer production, journalists and researchers, and finally, we investigate more or less occurrence of cited discourse, in what is concerned with different perspectives at communities that produce this kind of text. As theoretical background to our discussions we followed socio-historical perspective, its language and subject discourse conceptualizations. We did it mainly based on Bakhtin s works (1929; 1995; 2003). We were also based on theoretical discussions about discursive heterogeneity by Authier-Revuz (1990; 1998; 2004) and Maingueneau (1993; 2001). At analyzing the social dimensions of our data, we identified as relevant elements in the construction of the subjects (stories) the image that the writer (reporter) did/construct about his/her interlocutor as well as the use of different strategies, for example: the text produced by the journalists frequently use of direct discourse forms, while texts produced by researchers are almost fulfilled by indirect discourse. Beside this, texts are different in their social voices that are in their discourse. In the case of text produced by journalist are predominant the discursive scene of the school agents: teachers, students, parents, among others. Otherwise, in the texts produced by researchers already-said utterances, that in their majority of times, come from scientific discourse

Indicadores de cálcio e de voltagem codificados geneticamente na detecção de potenciais de ação e inputs sinápticos em cultura de neurônios hipocampais

Recently, genetically encoded optical indicators have emerged as noninvasive tools of high spatial and temporal resolution utilized to monitor the activity of individual neurons and specific neuronal populations. The increasing number of new optogenetic indicators, together with the absence of comparisons under identical conditions, has generated difficulty in choosing the most appropriate protein, depending on the experimental design. Therefore, the purpose of our study was to compare three recently developed reporter proteins: the calcium indicators GCaMP3 and R-GECO1, and the voltage indicator VSFP butterfly1.2. These probes were expressed in hippocampal neurons in culture, which were subjected to patchclamp recordings and optical imaging. The three groups (each one expressing a protein) exhibited similar values of membrane potential (in mV, GCaMP3: -56 ±8.0, R-GECO1: -57 ±2.5; VSFP: -60 ±3.9, p = 0.86); however, the group of neurons expressing VSFP showed a lower average of input resistance than the other groups (in Mohms, GCaMP3: 161 ±18.3; GECO1-R: 128 ±15.3; VSFP: 94 ±14.0, p = 0.02). Each neuron was submitted to current injections at different frequencies (10 Hz, 5 Hz, 3 Hz, 1.5 Hz, and 0.7 Hz) and their fluorescence responses were recorded in time. In our study, only 26.7% (4/15) of the neurons expressing VSFP showed detectable fluorescence signal in response to action potentials (APs). The average signal-to-noise ratio (SNR) obtained in response to five spikes (at 10 Hz) was small (1.3 ± 0.21), however the rapid kinetics of the VSFP allowed discrimination of APs as individual peaks, with detection of 53% of the evoked APs. Frequencies below 5 Hz and subthreshold signals were undetectable due to high noise. On the other hand, calcium indicators showed the greatest change in fluorescence following the same protocol (five APs at 10 Hz). Among the GCaMP3 expressing neurons, 80% (8/10) exhibited signal, with an average SNR value of 21 ±6.69 (soma), while for the R-GECO1 neurons, 50% (2/4) of the neurons had signal, with a mean SNR value of 52 ±19.7 (soma). For protocols at 10 Hz, 54% of the evoked APs were detected with GCaMP3 and 85% with R-GECO1. APs were detectable in all the analyzed frequencies and fluorescence signals were detected from subthreshold depolarizations as well. Because GCaMP3 is the most likely to yield fluorescence signal and with high SNR, some experiments were performed only with this probe. We demonstrate that GCaMP3 is effective in detecting synaptic inputs (involving Ca2+ influx), with high spatial and temporal resolution. Differences were also observed between the SNR values resulting from evoked APs, compared to spontaneous APs. In recordings of groups of cells, GCaMP3 showed clear discrimination between activated and silent cells, and reveals itself as a potential tool in studies of neuronal synchronization. Thus, our results indicate that the presently available calcium indicators allow detailed studies on neuronal communication, ranging from individual dendritic spines to the investigation of events of synchrony in neuronal networks genetically defined. In contrast, studies employing VSFPs represent a promising technology for monitoring neural activity and, although still to be improved, they may become more appropriate than calcium indicators, since neurons work on a time scale faster than events of calcium may foresee

A novel mutation (Q40P) in PAX8 associated with congenital hypothyroidism and thyroid hypoplasia: Evidence for phenotypic variability in mother and child

Congenital hypothyroidism associated with thyroid hypoplasia can be caused by several genetic defects, including mutations in the TSH beta -subunit, the TSH receptor, the G(A)alpha -subunit, and the transcription factor PAX8. Four girls with sporadic congenital hypothyroidism and hypoplastic thyroid glands were analyzed for mutations in PAX8 and TTF2 (FKHL15). Mutations in the coding region of the TSH beta -subunit gene, the TSH receptor gene, and exons 8 and 9 of G(mu)alpha had been excluded previously. Serum TSH concentrations were 150 mU/liter or more, TG levels were within normal limits, and thyroid autoantibodies were absent. Technetium scintigraphies did not reveal the presence of thyroid tissue, but ultrasonography documented hypoplastic, normally located glands.One patient was found to harbor a heterozygous transversion 119A -->C in exon 3 of PAX8 replacing a conserved glutamine by proline in the paired box domain (Q40P). Analysis of her family members revealed that her mother, who has a thyroid gland of normal size and mild, adult-onset autoimmune hypothyroidism, is also heterozygous for this mutation. Functional analyses of the PAX8 Q40P mutation showed impaired binding to a PAX8 response element and absent transactivation of a thyroid peroxidase promoter luciferase reporter gene.These findings confirm the important role of PAX8 in the development of the thyroid, but they indicate that PAX8 gene mutations may have a variable penetrance or expressivity. The absence of mutations in the coding sequences of the analyzed genes in the three other patients supports the concept that the pathogenesis of congenital hypothyroidism associated with thyroid hypoplasia is diverse.

MKK3/6-p38 MAPK signaling is required for IL-1 beta and TNF-alpha-induced RANKL expression in bone marrow stromal cells

Coupled bone turnover is directed by the expression of receptor-activated NF-kappa B ligand (RANKL) and its decoy receptor, osteoprotegerin (OPG). Proinflammatory cytokines, such as interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) induce RANKL expression in bone marrow stromal cells. Here, we report that IL-1 beta and TNF-alpha-induced RANKL requires p38 mitogen-activating protein kinase (MAPK) pathway activation for maximal expression. Real-time PCR was used to assess the p38 contribution toward IL-1 beta and TNF-alpha-induced RANKL mRNA expression. Steady-state RANKL RNA levels were increased approximately 17-fold by IL-1 beta treatment and subsequently reduced similar to 70%-90% when p38 MAPK was inhibited with SB203580. RANKL mRNA stability data indicated that p38 MAPK did not alter the rate of mRNA decay in IL-1 beta-induced cells. Using a RANKL-luciferase cell line receptor containing a 120-kB segment of the 5' flanking region of the RANKL gene, reporter expression was stimulated 4-5-fold by IL-1 beta or TNF-alpha treatment. IL-1 beta-induced RANKL reporter expression was completely blocked with specific p38 inhibitors as well as dominant negative mutant constructs of MAPK kinase-3 and -6. In addition, blocking p38 signaling in bone marrow stromal cells partially inhibited IL-1 beta and TNF-alpha-induced osteoclastogenesis in vitro. Results from these studies indicate that p38 MAPK is a major signaling pathway involved in IL-1 beta and TNF-alpha-induced RANKL expression in bone marrow stromal cells.

The promoter of a gene encoding an isoflavone reductase-like protein in coffee (Coffea arabica) drives a stress-responsive expression in leaves

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Functional and Ultrastructural Analysis of Annexin A1 and Its Receptor in Extravasating Neutrophils during Acute Inflammation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)