Splicing Modulation as a Promising Therapeutic Strategy for Lysosomal Storage Disorders: The Mucopolysaccharidoses Example

This article belongs to the Special Issue New Insights into Lysosomal Storage Disorders and Other Rare Genetic Diseases.Over recent decades, the many functions of RNA have become more evident. This molecule has been recognized not only as a carrier of genetic information, but also as a specific and essential regulator of gene expression. Different RNA species have been identified and novel and exciting roles have been unveiled. Quite remarkably, this explosion of novel RNA classes has increased the possibility for new therapeutic strategies that tap into RNA biology. Most of these drugs use nucleic acid analogues and take advantage of complementary base pairing to either mimic or antagonize the function of RNAs. Among the most successful RNA-based drugs are those that act at the pre-mRNA level to modulate or correct aberrant splicing patterns, which are caused by specific pathogenic variants. This approach is particularly tempting for monogenic disorders with associated splicing defects, especially when they are highly frequent among affected patients worldwide or within a specific population. With more than 600 mutations that cause disease affecting the pre-mRNA splicing process, we consider lysosomal storage diseases (LSDs) to be perfect candidates for this type of approach. Here, we introduce the overall rationale and general mechanisms of splicing modulation approaches and highlight the currently marketed formulations, which have been developed for non-lysosomal genetic disorders. We also extensively reviewed the existing preclinical studies on the potential of this sort of therapeutic strategy to recover aberrant splicing and increase enzyme activity in our diseases of interest: the LSDs. Special attention was paid to a particular subgroup of LSDs: the mucopolysaccharidoses (MPSs). By doing this, we hoped to unveil the unique therapeutic potential of the use of this sort of approach for LSDs as a whole.This research was partially funded by the FCT (FCT/PTDC/BBB-BMD/6301/2014 and EXPL/BTM-SAL/0659/2021), the Portuguese Society for Metabolic Disorders (Sociedade Portuguesa de Doenças Metabólicas, SPDM—Bolsa SPDM de apoio à investigação Dr Aguinaldo Cabral 2018 (2019DGH1629/SPDM2018I&D) and 2019 (2020DGH1834)), the Sanfilippo Children’s Foundation (SCF Incubator Grant 2019: 2019DGH1656/SCF2019I&D) and the MPS Society (2019DGH1642).info:eu-repo/semantics/publishedVersio

A evasão nas turmas de EJA : desafios para professores e alunos

Trabalho de Conclusão de Curso (especialização)—Universidade Aberta do Brasil, Universidade de Brasília, Faculdade de Educação, Curso de Especialização em Educação na Diversidade e Cidadania, com Ênfase em EJA, 2010.O presente projeto, “Evasão nas turmas de EJA: Desafios para professores e alunos” foi construído a partir da necessidade de intervenção verificada na Escola Municipal Valparaiso II, visando acima de tudo, senão resolver, amenizar situações-problema e paralelamente, apontar caminhos que possam direcionar novas práticas, no que diz respeito à redução do elevado índice de evasão que se constata a cada semestre letivo nas turmas de EJA. Tal fato apresenta-se como um grande desafio para os professores que buscam compreender que fatores levam os alunos a se evadirem e o que pode ser feito para reverter essa realidade, priorizando o diálogo e atividades conjuntas de professores, alunos e demais funcionários, dentro de uma perspectiva transdisciplinar, na tentativa de diminuir a evasão nas turmas de EJA

Neurological Disease Modeling Using Pluripotent and Multipotent Stem Cells: A Key Step towards Understanding and Treating Mucopolysaccharidoses

(This article belongs to the Special Issue Molecular Mechanisms in Lysosomal Storage Diseases: From Pathogenesis to Therapeutic Strategies 2.0)ReviewDespite extensive research, the links between the accumulation of glycosaminoglycans (GAGs) and the clinical features seen in patients suffering from various forms of mucopolysaccharidoses (MPSs) have yet to be further elucidated. This is particularly true for the neuropathology of these disorders; the neurological symptoms are currently incurable, even in the cases where a disease-specific therapeutic approach does exist. One of the best ways to get insights on the molecular mechanisms driving that pathogenesis is the analysis of patient-derived cells. Yet, not every patient-derived cell recapitulates relevant disease features. For the neuronopathic forms of MPSs, for example, this is particularly evident because of the obvious inability to access live neurons. This scenario changed significantly with the advent of induced pluripotent stem cell (iPSC) technologies. From then on, a series of differentiation protocols to generate neurons from iPSC was developed and extensively used for disease modeling. Currently, human iPSC and iPSC-derived cell models have been generated for several MPSs and numerous lessons were learnt from their analysis. Here we review most of those studies, not only listing the currently available MPS iPSC lines and their derived models, but also summarizing how they were generated and the major information different groups have gathered from their analyses. Finally, and taking into account that iPSC generation is a laborious/expensive protocol that holds significant limitations, we also hypothesize on a tempting alternative to establish MPS patient-derived neuronal cells in a much more expedite way, by taking advantage of the existence of a population of multipotent stem cells in human dental pulp to establish mixed neuronal and glial cultures.Funding: This work was partially supported by the Portuguese Society for Metabolic Disorders, SPDM (Bolsa SPDM de apoio à investigação Dr. Aguinaldo Cabral 2018; 2019DGH1629/SPDM2018I&D), Sanfilippo Children’s Foundation (2019DGH1656/SCF2019I&D), and Fundação para a Ciência e a Tecnologia, FCT: EXPL/BTM-SAL/0659/2021; UIDB/00211/2020—Centro de Estudos de Ciência Animal/Center for the Study of Animal Science; LA/P/0059/2020—Laboratório Associado para Ciência Animal e Veterinária/Associate Laboratory for Animal and Veterinary Sciences.info:eu-repo/semantics/publishedVersio

Non-canonical Wnt signaling regulates junctional mechanocoupling during angiogenic collective cell migration

© 2019, Carvalho et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.Morphogenesis of hierarchical vascular networks depends on the integration of multiple biomechanical signals by endothelial cells, the cells lining the interior of blood vessels. Expansion of vascular networks arises through sprouting angiogenesis, a process involving extensive cell rearrangements and collective cell migration. Yet, the mechanisms controlling angiogenic collective behavior remain poorly understood. Here, we show this collective cell behavior is regulated by non-canonical Wnt signaling. We identify that Wnt5a specifically activates Cdc42 at cell junctions downstream of ROR2 to reinforce coupling between adherens junctions and the actin cytoskeleton. We show that Wnt5a signaling stabilizes vinculin binding to alpha-catenin, and abrogation of vinculin in vivo and in vitro leads to uncoordinated polarity and deficient sprouting angiogenesis in Mus musculus. Our findings highlight how non-canonical Wnt signaling coordinates collective cell behavior during vascular morphogenesis by fine-tuning junctional mechanocoupling between endothelial cells.Research was supported by European Research Council starting grant (679368), the H2020-Twinning grant (692322), the Fundação para a Ciência e a Tecnologia funding (grants: IF/00412/2012; EXPL-BEX-BCM-2258–2013; PRECISE-LISBOA-01–0145-FEDER-016394; UID/BIM/50005/2019, a project funded by Fundação para a Ciência e a Tecnologia (FCT)/ Ministério da Ciência,Tecnologia e Ensino Superior (MCTES) through Fundos do Orçamento de Estado; and a grant from the Fondation Leducq (17CVD03); and personal fellowships: BD/52224/2013​​ to JRC, BD/105856/2014 to PB, and BD/128375/2017 to CF) and LISBOA-01–0145-FEDER-007391, project cofunded by FEDER, through POR Lisboa 2020 - Programa Operacional Regional de Lisboa, PORTUGAL 2020, and Fundação para a Ciência e a Tecnologia.info:eu-repo/semantics/publishedVersio

The disease modelling value of baby teeth: A new way to unlock knowledge about a special group of genetic disorders

Mucopolysaccharidoses (MPS), are a group of genetic, metabolic, and rare diseases investigated since the early years of the 20th century. One of the first steps to collect information about the underlying mechanisms of those disorders is the development and analysis of in vitro models. Furthermore, those models provide an appropriate platform for the evaluation of future therapeutics. Among all the possible disease cell models, patient-derived ones are those which allow us to get better disease insights. However, finding the best cell type that recapitulates diseaserelevant features is not always easy: two systems largely involved in MPS pathology are the brain and the musculoskeletal ones, which reflects an issue once both are hard to access. Here, our main goal is to establish an innovative non-invasive method to generate disease-relevant cell models from stem cells from deciduous (baby) teeth (SHED), which may then be differentiated into our MPS-target cell lines. So far, we have already implemented and optimized the protocol for collection, isolation, establishment and cryopreservation of those stem cells. Then, our rationale is simple: for those obtained from MPS patients suffering from multisystemic disease with marked musculoskeletal alterations, we are using a chondrogenesis differentiation protocol. For those derived from patients with neurological pathology, we will establish mixed neuronal/glial cultures. As soon as we can get the SHED-derived differentiated cells, various cellular and molecular processes from our target disorders may be unveiled and used as a target for possible future therapeutics. Acknowledgements This work is partially supported by the Portuguese Society for Metabolic Disorders (SPDM - Bolsa SPDM de apoio à investigação Dr. Aguinaldo Cabral 2018;2019DGH1629/SPDM2018I&D), Sanfilippo Children's Foundation (2019DGH1656/SCF2019I&D) and FCT (EXPL/BTM-SAL/0659/2021).This work is partially supported by the Portuguese Society for Metabolic Disorders (SPDM - Bolsa SPDM de apoio à investigação Dr. Aguinaldo Cabral 2018;2019DGH1629/SPDM2018I&D), Sanfilippo Children's Foundation (2019DGH1656/SCF2019I&D) and FCT (EXPL/BTM-SAL/0659/2021),N/

Emoções de mães de bebês prematuros: a perspectiva de profissionais da saúde

This article analyzed the emotional aspects of mothers of premature babies and the mother- baby interactions from the testimonials of health professionals of a public maternity. Five participants of the Kangaroo Program underwent a series of interviews, during the hospitalization of the babies. From what was said, three categories were analyzed- mother’s emotions concerning the time when the baby was given birth, in which the participants realized how stressing giving birth to a premature baby can be and how anxious mothers feel, also scared and hopeless when faced to such frailty and instability of the baby. Secondly, emotional aspects of mothers during the hospitalization of the baby, emerged feelings of anxiety, fear, insecurity and overcoming difficulties and to the interviewed, in this period, mothers go through an unstable mood that varies from an anxious and insecure personality to a hopeful and secure one. Finally, in the category of interactions between mother-baby in the Kangaroo mother care method, all the participants affirmed that this method facilitates such interactions.Key words: health staff, mothers of premature babies, Kangaroo mother care method.Este artigo analisou aspectos emocionais de mães de prematuros e as interações mãe-bebê a partir das falas de profissionais da saúde de uma maternidade pública. Cinco integrantes do Programa Canguru, foram submetidos a um roteiro de entrevista, durante a hospitalização dos bebês. Das falas, três categorias foram analisadas: Emoções maternas diante do nascimento do filho, na qual os participantes perceberam o nascimento prematuro como estressante e ansiogênico para as mães, gerando susto, medo, ansiedade e desesperança face à fragilidade e à instabilidade do bebê. Na segunda, Aspectos emocionais maternos durante a hospitalização do bebê, emergiram sentimentos como ansiedade, medo, insegurança e superação de dificuldades. Para os entrevistados, nesse período, as mães vivenciam um processo emocional que oscila da ansiedade e da insegurança a um estado de esperança e segurança. Por fim, na categoria Interações entre a díade mãe-bebê no método canguru, todos os participantes afirmaram que esse método facilita tais interações.Palavras-chave: equipe de saúde, mães de prematuros, método canguru

ANÁLISE PRELIMINAR DA PAISAGEM COSTEIRA DO LITORAL DO ESTADO DO RIO GRANDE DO NORTE, NE DO BRASIL.

Esta pesquisa visa fazer um levantamento da atual realidade da linha de costa do litoral do Estado do Rio Grande do Norte, no Nordeste do Brasil, entre as praias de Sagi (divisa do RN/ PB) e a praia de Tibau (divisa do RN/CE), identificando os pontos que estão em processos mais erosivos, e buscando uma solução para cada realidade de cada praia pesquisada. Nesta etapa, definiremos os nomes de todas as praias do nosso litoral, bem como situaremos os pontos que já são críticos, pelo menos do ponto de vista erosional e atuais. Neste processo identificamos mudanças na paisagem costeira provenientes tanto da ação antrópica, como da ação natural das praias. Pretendemos com isso, fornecer as referidas prefeituras litorâneas do Estado, os resultados e propostas técnicas, indicando soluções possíveis para a recuperação da paisagem costeira

LICENCIAMENTO AMBIENTAL NOS ESTADOS BRASILEIROS: OS PROCEDIMENTOS SERIAM OS MESMOS PARA UMA RODOVIA?

Highways consist of structures whose importance is attributed to the ability to provide circulation between places of people and cargo. This typology of enterprises affects the environment differently according to the characteristics of the project and the environmental in which they are inserted. Therefore, they are considered activities potentially causing environmental degradation, and therefore, subject to Environmental Licensing (EL) by the competent environmental agency. Faced with the legal possibility of different procedures for each state, the article aims to compare the state Environmental Licensing process of a hypothetical state highway with a length of 50 km in seven Brazilian states: Bahia, Espírito Santo, Minas Gerais, Paraná, Rio de Janeiro, Rio Grande do Sul and Tocantins. The methodology applied involved documental and content analysis of legislation and the website of licensing agencies, in order to gather and compare information and data of EL process. In each state, the environmental agency, the state environmental council and the legislation guiding the EL process were raised, as well as the need for an Environmental Impact Assessment (EIA) and an Environmental Impact Study (EIS) for the hypothetical highway. A difference can be noted in the classification of the polluting potential and the size of the activity in the different states. However, all states point to the need to carry out the EIA for the analysis of the project's environmental feasibility. However, not all of them adhere to the EIS as an environmental study, leaving the possibility of more simplified studies. We hope that the comparisons and discussions carried out in this article can serve as a guide for the process of improving Environmental Licensing and Environmental Impact Assessment in Brazil.Rodovias consistem em estruturas cuja importância é atribuída à capacidade de propiciar a circulação entre lugares de pessoas e carga. Empreendimentos desta tipologia impactam o ambiente de forma diferenciada de acordo com as características do projeto e do meio em que estão inseridos. Por isso, são consideradas atividades potencialmente causadoras de degradação ambiental, sendo assim, passíveis de Licenciamento Ambiental (LA) junto ao órgão ambiental competente. Frente à possibilidade legal de procedimentos diferenciados para cada estado, o artigo tem como objetivo comparar o processo de LA estadual de uma rodovia estadual hipotética com extensão de 50 km em sete estados brasileiros: Bahia, Espírito Santo, Minas Gerais, Paraná, Rio de Janeiro, Rio Grande do Sul e Tocantins. A metodologia aplicada envolveu análise documental e de conteúdo de legislações e da página eletrônica dos órgãos licenciadores estaduais, visando levantar e comparar informações e dados relacionados ao processo de LA. Levantou-se em cada estado o órgão ambiental, o conselho estadual de meio ambiente e a legislação norteadora do LA, assim como, anecessidade da Avaliação de Impacto Ambiental (AIA) e do Estudo de Impacto Ambiental (EIA) para a rodovia hipotética. Pode-se notar uma diferença na classificação do potencial poluidor e do porte da atividade nos diferentes estados, mas todos eles apontam para a necessidade de realização da AIA para a análise da viabilidade do empreendimento, porém, nem todos se atêm ao EIA como estudo ambiental, deixando possibilidade de estudos mais simplificados. Espera-se que as comparações e discussões realizadas neste artigo possam servir como orientação para o processo de melhoria do Licenciamento e Avaliação de Impacto Ambiental no Brasil.

PERFIL DE PESSOAS COM E SEM COMORBIDADES ACOMETIDAS POR REAÇÕES HANSÊNICAS

This study aimed to identify clinical andepidemiological characteristics in patients with and withoutcomorbidities from leprosy and leprosy reactions. It isquantitative, descriptive and correlational research, undertakenthrough 65 interviews in a specialized center in Rio Grande doNorte, Brazil, with data collected between October 2013 andMarch 2014. The men represented most of the group withoutcomorbidities, while women and older adults represented thegroup with comorbidities. High frequencies in the two groupspoint to the Lepromatous form as the most common, andto some degree of disability. There was positive correlationbetween number of other drugs versus other comorbiditiesand negative correlation between number of people wholive with the interviewee versus the number of comorbiditieswhich the interviewee has. Late diagnosis was evidenced dueto high frequencies of disabilities, as well as groups at risk forleprosy reactions and comorbidities, made up of women, olderadults, people with low educational level, people who did notwork and those who used a higher number of medications.Estudio cuyo propósito fue identificar característicasclínicas y epidemiológicas en pacientes con y sin comorbilidadespor enfermedad de Hansen y reacciones a esa enfermedad.Investigación cuantitativa, descriptiva y correlacional, realizadapor 65 entrevistas en un centro de referencia. Los datos fueronobtenidos entre octubre de 2013 y marzo de 2014. Había máshombres en grupo sin comorbilidades, en cuanto las mujeres ylos ancianos representaron más el grupo con comorbilidades.Altas frecuencias en los dos grupos apuntan para la formaVirchowiana como la con más incidencia y para algun gradode incapacidad. Hubo correlación positiva entre cantidad deotros medicamentos versus otras comorbilidades y negativaentre cantidad de personas que viven con el entrevistadoversus número de comorbilidades que este presenta. Fueronevidenciados diagnósticos tardíos en función de altasfrecuencias de incapacidades, así como grupos de riesgopara reacciones hansénicas y comorbilidades, compuesto pormujeres, ancianos, personas con poca escolaridad, que notrabajaban y que utilizaban mayor cantidad de medicamentos.Objetivou-se identificar características clínicas e epidemiológicas em pacientes com e sem comorbidadespor hanseníase e reações hansênicas. Pesquisa quantitativa, descritiva e correlacional, realizada por 65 entrevistasem um centro de referência potiguar, com coleta de dados entre outubro de 2013 e março de 2014. Os homensrepresentaram mais o grupo sem comorbidades, enquanto as mulheres e os idosos o grupo com comorbidades.Altas frequências nos dois grupos apontam para a forma Virchowiana como mais incidente e para algum grau deincapacidade. Houve correlação positiva entre quantidade de outros medicamentos versus outras comorbidadese negativa entre quantidade de pessoas que moram com o entrevistado versus número de comorbidades que estepossui. Foram evidenciados diagnósticos tardios em virtude de altas frequências de incapacidades, bem como gruposde risco para reações hansênicas e comorbidades, composto por mulheres, idosos, pessoas com baixa escolaridade,que não trabalhavam e que utilizavam maior quantidade de medicamentos