Structural and functional basis for p38-MK2 activated Rsk signalling in Toll-Like receptor-stimulated dendritic cells

Rsk kinases play important roles in several cellular processes such as proliferation, metabolism, and migration. Until recently, Rsk activation was thought to be exclusively initiated by Erk1/2, but in dendritic cells (DC) Rsk is also activated by p38 mitogen-activated protein (MAP) kinase via its downstream substrates, MK2/3. How and why this noncanonical configuration of the MAP kinase pathway is adopted by these key immune cells are not known. We demonstrate that the Erk1/2-activated C-terminal kinase domain of Rsk is dispensable for p38-MK2/3 activation and show that compared with fibroblasts, a greater fraction of p38 and MK2/3 is located in the cytosol of DC prior to stimulation, suggesting a partial explanation for the operation of the noncanonical pathway of Rsk activation in these cells. p38/MK2/3-activated Rsk phosphorylated downstream targets and is physiologically important because in plasmacytoid DC (pDC) stimulated with Toll-like receptor 7 (TLR7) agonists, Erk1/2 activation is very weak relative to p38. As a result, Rsk activation is entirely p38 dependent. We show that this unusual configuration of MAP kinase signaling contributes substantially to production of type I interferons, a hallmark of pDC activation

p90Rsk is not involved in cytostatic factor arrest in mouse oocytes

Vertebrate oocytes arrest in metaphase of the second meiotic division (MII), where they maintain a high cdc2/cyclin B activity and a stable, bipolar spindle because of cytostatic factor (CSF) activity. The Mos–MAPK pathway is essential for establishing CSF. Indeed, oocytes from the mos−/− strain do not arrest in MII and activate without fertilization, as do Xenopus laevis oocytes injected with morpholino oligonucleotides directed against Mos. In Xenopus oocytes, p90Rsk (ribosomal S6 kinase), a MAPK substrate, is the main mediator of CSF activity. We show here that this is not the case in mouse oocytes. The injection of constitutively active mutant forms of Rsk1 and Rsk2 does not induce a cell cycle arrest in two-cell mouse embryos. Moreover, these two mutant forms do not restore MII arrest after their injection into mos−/− oocytes. Eventually, oocytes from the triple Rsk (1, 2, 3) knockout present a normal CSF arrest. We demonstrate that p90Rsk is not involved in the MII arrest of mouse oocytes

SENSIBILIZAÇÃO DOS ALUNOS DA EDUCAÇÃO BÁSICA QUANTO À CONSCIENTIZAÇÃO AMBIENTAL

A Educação Ambiental é algo que está presente durante toda a nossa vida, podendo então ser trabalhada na escola desde muito cedo para prática da sensibilização quanto às questões ambientais. A natureza vem sendo agredida pelos seres humanos, e a possível solução para sanar essa destruição é a educação. Assim o ensino pode ser pautado em valores sociais tais como a sustentabilidade, responsabilidade, ética e também quanto a questões da cidadania, buscando a formação integral do ser humano num convívio pacífico com o meio ambiente em todas as suas instâncias. É necessário que esta prática seja realmente efetiva na vida do educando, pois a mesma se dá nas vivências e nas atitudes cotidianas de cada um, onde está impregnado o desperdício, descuido e a falta de responsabilidade para com a natureza e seus recursos naturais, com a água, o ar e o solo, estes tão importantes para a vida na Terra. Uma forma para o processo de sensibilização quanto à conscientização ambiental é a reciclagem do lixo que produzimos – papel, plástico, metal, vidro, orgânico, dentre outros. Para o ensino pautado nas questões ambientais acontecer efetivamente nas escolas, este pode ser abordado de maneira interdisciplinar, ou seja, integrar todas as disciplinas numa só perspectiva. Portanto, o docente (regente) é um dos responsáveis por um planejamento mais dinâmico junto aos professores de outros componentes curriculares, a fim de obter um estudo mais abrangente e significativo para os seus alunos, o que se tornará uma prática e mudança futura de nossa sociedade

The PDK1-Rsk signaling pathway controls Langerhans cell proliferation and patterning

Langerhans cells (LC), the dendritic cells of the epidermis, are distributed in a distinctive regularly spaced array. In the mouse, the LC array is established in the first few days of life from proliferating local precursors, but the regulating signaling pathways are not fully understood. We found that mice lacking the kinase phosphoinositide-dependent kinase 1 selectively lack LC. Deletion of the phosphoinositide-dependent kinase 1 target kinases, ribosomal S6 kinase 1 (Rsk1) and Rsk2, produced a striking perturbation in the LC network: LC density was reduced 2-fold, but LC size was increased by the same magnitude. Reduced LC numbers in Rsk1/2?/? mice was not due to accelerated emigration from the skin but rather to reduced proliferation at least in adults. Rsk1/2 were required for normal LC patterning in neonates, but not when LC were ablated in adults and replaced by bone marrow–derived cells. Increased LC size was an intrinsic response to reduced LC numbers, reversible on LC emigration, and could be observed in wild type epidermis where LC size also correlated inversely with LC density. Our results identify a key signaling pathway needed to establish a normal LC network and suggest that LC might maintain epidermal surveillance by increasing their “footprint” when their numbers are limite

Especialização do Ministério Público nos Estados na Temática Educação: Extensão, Características e Dinâmica Institucional

O trabalho investiga a especialização do Ministério Público (MP) nos vinte e seis estados, na temática Educação, um fenômeno que decorre tanto de obrigações legais como de decisões institucionais de cada estado, em parte relacionadas a orientações nacionais emanadas do Conselho Nacional do Ministério Público (CNMP). No presente estudo, a unidade de análise é cada órgão especializado do MP dos Estados, classificados conforme o grau de especialização, o tipo, as atribuições funcionais que reúnem e a amplitude de atuação, dentre outros elementos. As fontes são as normas de organização, portais de transparência e entrevistas complementares aplicadas aos membros e servidores, quando necessário. Como resultados, propomos um modelo de classificação e análise da especialização temática na área da educação e analisamos os achados à luz da literatura estabelecida. Com isso, o trabalho permite avançar na compreensão teórica e no conhecimento da realidade institucional do MP, um dos principais protagonistas do sistema de justiça no marco dos direitos sociais da Constituição de 1988, que lhe assegurou poderes e autonomias sem precedentes. A atuação do MP, neste marco, impacta diretamente o processo de políticas públicas e, como consequência, interfere nas disputas políticas sobre a distribuição de bens e direitos na sociedade brasileira. Estudar a especialização do MP permite desvelar também as orientações político-institucionais do MP que, assim definidas, orientam a atuação de seus membros em direção a temáticas tidas como prioritárias ou, ao menos, entendidas como passíveis de um direcionamento específico via política institucional

Reducing the Clinical and Public Health Burden of Familial Hypercholesterolemia A Global Call to Action

Q1Q1Artículo completoE1-E13IMPORTANCE Familial hypercholesterolemia (FH) is an underdiagnosed and undertreated genetic disorder that leads to premature morbidity and mortality due to atherosclerotic cardiovascular disease. Familial hypercholesterolemia affects 1 in 200 to 250 people around the world of every race and ethnicity. The lack of general awareness of FH among the public and medical community has resulted in only 10% of the FH population being diagnosed and adequately treated. The World Health Organization recognized FH as a public health priority in 1998 during a consultation meeting in Geneva, Switzerland. The World Health Organization report highlighted 11 recommendations to address FH worldwide, from diagnosis and treatment to family screening and education. Research since the 1998 report has increased understanding and awareness of FH, particularly in specialty areas, such as cardiology and lipidology. However, in the past 20 years, there has been little progress in implementing the 11 recommendations to prevent premature atherosclerotic cardiovascular disease in an entire generation of families with FH. OBSERVATIONS In 2018, the Familial Hypercholesterolemia Foundation and the World Heart Federation convened the international FH community to update the 11 recommendations. Two meetings were held: one at the 2018 FH Foundation Global Summit and the other during the 2018 World Congress of Cardiology and Cardiovascular Health. Each meeting served as a platform for the FH community to examine the original recommendations, assess the gaps, and provide commentary on the revised recommendations. The Global Call to Action on Familial Hypercholesterolemia thus represents individuals with FH, advocacy leaders, scientific experts, policy makers, and the original authors of the 1998 World Health Organization report. Attendees from 40 countries brought perspectives on FH from low-, middle-, and high-income regions. Tables listing country-specific government support for FH care, existing country-specific and international FH scientific statements and guidelines, country-specific and international FH registries, and known FH advocacy organizations around the world were created. CONCLUSIONS AND RELEVANCE By adopting the 9 updated public policy recommendations created for this document, covering awareness; advocacy; screening, testing, and diagnosis; treatment; family-based care; registries; research; and cost and value, individual countries have the opportunity to prevent atherosclerotic heart disease in their citizens carrying a gene associated with FH and, likely, all those with severe hypercholesterolemia as well

TheFamily of RSK Proteins (Genetic aspects of coffin-lowry syndrome, involving RSK2, and functional studies on RSK2 and two related proteins, RSK1 and RSK3)

Les retards mentaux liés au chromosome X peuvent être syndromiques (MRXS) ou non-syndromiques (MRX). Dans le cas du syndrome de Coffin-Lowry (CLS), une forme de MRXS, le retard mental est associé à des anomalies notamment squelettiques. Des mutations de perte de fonction de RSK2 sont la cause de ce syndrome. Chez l'homme, RSK2 fait partie d'une famille de quatre kinases de la voie Ras/ERK-MAPK. Nous avons identifié une mutation dans le gène RSK2 dans une famille MRX, ce qui élargit le spectre phénotypique des mutations dans ce gène. Nous avons aussi montré que les techniques de western blot et de test kinase in vitro peuvent être utilisées pour le diagnostic moléculaire de CLS. Ces techniques associées à une recherche de mutations dans le promoteur du gène RSK2, ont suggéré une probable hétérogénéité génétique en ce qui concerne ce syndrome. Elles ont aussi permis l'identification de deux mutations inhabituelles d'épissage que nous avons étudié en détail. Afin de mieux comprendre les fonctions des RSKs, nous avons généré des anticorps reconnaissant spécifiquement les protéines RSK1, 2 et 3. Ces anticorps ont permis de déterminer qu' alors que RSK3 est présente de manière uniforme dans le cytoplasme et le noyau des cellules, RSK1 est principalement détectée dans des zones bien délimitées du noyau, les "speckles". Nous avons utilisé ces anticorps ainsi que les techniques de northern blot et d'hybridation in situ afin de déterminer l'expression tissulaire des RSKs. RSK1, 2 et 3 étaient toutes exprimées dans un grand nombre de tissus. Cependant, uniquement RSK2 est fortement exprimée dans certains régions du cerveau adulte impliquées dans des processus de mémoire, ce qui pourrait expliquer le déficit cognitif observé chez les patients CLS. Enfin, nous avons généré des souris invalidées pour l'expression des gènes Rsk1 et Rsk3, qui avec les animaux invalidés pour l'expression de Rsk2, seront utiles pour l'identification des fonctions spécifiques et redondantes des RSKs.Mental retardation (MR) affects 1 to 1.5% of the population. X-linked mental retardation is divided into two classes: syndromic (MRXS) and nonsyndromic or nonspecific (MRX). The Coffin-Lowry syndrome (CLS) is a form of MRXS in which the cognitive deficit is associated to growth retardation and skeletal malformations. CLS is caused by loss of function mutations in the RSK2 gene encoding the RSK2 protein. In humans, RSK2 is member of a family of four highly related serine/threonine kinases (RSK1-4) acting in the Ras/ERK-MAPK signaling pathway and involved in various cellular processes. We found a mutation in the RSK2 gene in an MRX family, extending the phenotypic variability in patients carrying RSK2 mutations. We also showed that western blotting and in vitro kinase assays are efficient tests for molecular diagnosis of CLS. These tests along with a high scale mutational screening in the promoter region of RSK2, indicated that genetic heterogeneity in CLS should not be excluded. Western blotting allowed also the identification of two unusual splicing mutations that were studied in detail. To better understand the functions of RSK proteins, we generated polyclonal antibodies recognizing specifically RSK1, 2 and 3. These antibodies were used to determine that whereas RSK3 was uniformly distributed in the cytoplasmic and nuclear compartments, RSK1 was mainly detected in nuclear speckles, suggesting a putative role of RSK1 in splicing processes. We have also used these antibodies, as well as northern blotting and in situ hybridization, to study the tissue expression of RSKs. RSK1, 2 and 3 were all widely expressed. However, only RSK2 was detected in some brain areas involved in memory processes, providing a possible explanation for the cognitive deficit observed in CLS patients. Finally, we have generated Rsk1 and Rsk3 knockout mice which will be useful, along with the Rsk2 knockout animals, for the identification of specific as well as redundant functions of RSKs.STRASBOURG-Sc. et Techniques (674822102) / SudocSudocFranceF

Identification de nouveaux gènes de retard mental par la caractérisation de translocations (X;Autosome) (KIAA1202 et CDKL3, deux nouveaux gènes candidats pour le retard mental)

Les retards mentaux (RM) concernent 1 à 2% de la population et représentent un véritable problème de santé publique. Ils se définissent comme une limitation significative à la fois du fonctionnement intellectuel général et des capacités d adaptation. Ils sont divisés en deux groupes : les RM syndromiques (RM associé à d autres signes cliniques) et les RM non syndromiques (RM isolé). L objectif de ma thèse était d identifier de nouveaux gènes responsables de RM lié au chromosome X (RMLX) non syndromiques et de comprendre leur implication dans le RM. Pour cela, nous avons étudié plusieurs patientes présentant une translocation (X;Autosome) associée à un RM. Pour deux patientes présentant un RM associé à une t(X;19) ou une t(X;8), nous avons mis en évidence l interruption du même gène, KIAA1202, au niveau du point de cassure sur le chromosome X. Nous avons également identifié une mutation faux sens dans une famille présentant le RMLX syndromique Stocco dos Santos. Des analyses complémentaires suggèrent que KIAA1202 aurait un rôle dans le fonctionnement et/ou le développement du cerveau. La protéine Shroom4 codée par le gène KIAA1202 est un nouveau membre de la famille de protéines APX/Shroom et interviendrait dans le remodelage du cytosquelette d actine. Pour les autres patientes analysées, aucun gène n était directement interrompu par la translocation sur le chromosome X. Pour la patiente MRX30513 présentant un RM associé à une t(X;5), nous avons mis en évidence l inactivation fonctionnelle d un gène autosomique, CDKL3, au niveau du point de cassure sur le chromosome 5. Des analyses d expression chez la souris suggèrent que Cdkl3 aurait un rôle dans le cerveau. Pour trois autres patientes, le point de cassure autosomique est en cours de caractérisation. Enfin, pour la dernière patiente étudiée, nous avons observé une délétion en plus de la translocation. En conclusion, cette stratégie a permis d identifier deux excellents gènes candidats pour le RM : KIAA1202 et CDKL3.Mental retardation (MR) affects 1 to 2% of the population and thus represents an important public health problem. MR is defined by a subaverage intellectual functioning associated with limitation in adaptive skills. Generally, MR is subdivided into two groups: syndromic mental retardation (MR associated with additional clinical symptoms) and non syndromic mental retardation (isolated MR). The objective of my thesis work was to identify new genes responsible for non syndromic X-linked mental retardation (XLMR) and to understand their involvement in the disease. For that purpose, we have performed a study on several females patients presenting mental retardation associated with a (X;Autosome) translocation. For two patients presenting mental retardation associated with a (X;19) or a (X;8) translocation, we highlighted the disruption of the same gene, KIAA1202, by the translocation breakpoint on chromosome X. We also identified a missense mutation in one family presenting the Stocco dos Santos XLMR syndrome. Further studies suggested that KIAA1202 would have a role in brain development and/or functioning. The protein Shroom4 encoded by the KIAA1202 gene is a new member of the APX/Shroom proteins family and would be involved in the actin cytoskeleton remodeling. For the others patients analysed, no gene was directly disrupted by the translocation on the X chromosome. For patient MRX30513, presenting a mental retardation associated with a (X;5) translocation, we revealed functional inactivation of an autonomal gene, CDKL3, by the breakpoint on chromosome 5. Expression analyses in mouse suggested that Cdkl3 would have a role in brain. For three others patients, the characterisation of the breakpoint on the autosome is in progress. Finally, for the last patient, we observed a deletion in addition to the translocation. In conclusion, this strategy allowed us to identify two excellent new mental retardation candidate genes: KIAA1202 and CDKL3.STRASBOURG-Sc. et Techniques (674822102) / SudocSudocFranceF

Identification des déficits moléculaires, cellulaires et cognitifs chez le modèle souris du retard mental causé par la mutation du gène Rsk2

Le syndrome de Coffin-Lowry (CLS), une déficience intellectuelle liée à l'X, est causée par des mutations du gène RPS6KA3 codant pour la kinase RSK2 régulée par les facteurs de croissance.Pour comprendre les conséquences du déficit en RSK2 dans l'hippocampe nous avons effectué une comparaison des profils d'expression génique d'hippocampes de souris Rsk2-KO et WT. Elle a révélé l'expression différentielle de 100 gènes, codant pour des protéines agissant dans diversprocessus biologiques. Nous avons analysé les conséquences de la dérégulation de l'un de ces gènes Gria2 codant pour GluR2, une sous-unité du récepteur glutamate AMPA. Un niveau d'expression doublé de GluR2 a été relevé dans l'hippocampe des souris Rsk2-KO et les études électrophysiologiques y ont révélé une réduction des transmissions AMPAR et NMDAR. L activité de ERK1/2 était aussi anormalement augmentée dans l'hippocampe des souris Rsk2-KO, ainsi que leniveau de P-Sp1. Ensemble, mes résultats ont suggéré que la surexpression de GluR2 dans les neurones déficients en RSK2, était causée par une augmentation de l'activité transcriptionnelle de Sp1 sur le gène Gria2, qui, elle-même, est le résultat de l augmentation anormale de l activité de ERK1 / 2.Coffin Lowry Syndrome (CLS), an X-linked form of intellectual disability, is caused by mutations of the RPS6KA3 gene encoding the growth factor regulated kinase RSK2. To understand the consequences of RSK2 deficiency in the hippocampus we performed a comparison of the hippocampal gene expression profiles from Rsk2-KO and WT mice. It revealed differentialexpression of 100 genes, encoding proteins acting in various biological pathways. We further analyzed the consequences of deregulation of one of these genes, Gria2 encoding GluR2, a subunit of the glutamate AMPAR. An abnormal two-fold increased expression of GluR2 was found in the hippocampus of Rsk2-KO mice. Electrophysiology studies showed a reduction of basal AMPAR and NMDAR mediated transmission, in the hippocampus of Rsk2-KO mice. Activity of ERK1/2 was also abnormally increased in the adult hippocampus of Rsk2-KO mice. P-Sp1 level was also significantly higher in RSK2 deficient cells. Together, my results suggested that over expression of GluR2 in RSK2 deficient cells, is caused by increased Sp1 transcriptional activity on the Gria2 gene, which, itself, is the result of ERK1/2 increased signaling.STRASBOURG-Bib.electronique 063 (674829902) / SudocSudocFranceF

A syndromic form of X-linked mental retardation: the Coffin-Lowry syndrome

Coffin-Lowry syndrome (CLS, OMIM 303600) is an X-linked inherited disorder characterised in male patients by growth and psychomotor retardation, hypotonia and progressive skeletal changes. Typically, male patients are of short stature and exhibit a characteristic coarse face with a prominent forehead, orbital hypertelorism, downslanting palpebral fissures, thick lips, a thick nasal septum with anteverted nares, and irregular or missing teeth. Their large and soft hands with lax skin and tapering fingers, are usually a strong diagnostic feature. Some patients present with additional complications including, sensorineural deafness, seizures, drop episodes and cardiac disease. There is some variability in the mental development of affected males, but most of the males who receive appropriate care appear to be moderately affected. A majority of carrier females have only minimal findings (mild facial coarsening, tapering fingers and obesity). Early diagnosis of CLS is essential for proper management, including survey of some specific complications already mentioned, and for genetic counselling. Establishing the diagnosis in very young children is often much more difficult than in older patients since physical characteristics are milder and not specific. Loss of function mutations in the gene encoding the growth-factor induced protein kinase ribosomal S6 kinase are responsible for CLS. A mutation has been detected in about 50% of patients with clinical features suggestive of CLS, and over 80 different mutations have so far been identified. They are distributed throughout the gene, the vast majority being unique to single families and a high proportion appear to be de novo events. Some missense mutations are associated with milder phenotypes. In one family, a missense mutation was associated solely with mild mental retardation and no other clinical feature. CONCLUSION: Coffin-Lowry syndrome is a well characterised entity and a detailed clinical examination usually allows diagnosis. However, recognising it in very young children is often difficult since physical characteristics are mild and not specific. In addition, most cases are sporadic. Screening for ribosomal S6 kinase mutations is essential in most cases to confirm the diagnosis as well as for genetic counselling