Emerging Roles of microRNAs in Cystic Fibrosis — From Pathogenesis to Development of New Therapies

As essential components of the regulatory system of gene expression, microRNAs (miRNAs) have been shown to influence development, severity, prognosis, and/or progression of a variety of inherited diseases. Differential expression studies have evidenced an impact of miRNAs on lung disease development in chronic obstructive pulmonary disease (COPD), asthma, lung inflammation, consequences of smoke exposure and airway allergy in human and in animal models of the diseases. Recent clinical and cell-based studies have revealed specific alterations of miRNA expression in cystic fibrosis (CF). Here we critically review the major findings concerning altered miRNA expression in CF airway epithelium, in particular with respect to CF transmembrane conductance regulator (CFTR) expression, innate immunity, and epithelial differentiation. Finally, we explore strategies to exploit these changes with the aim of innovative therapeutic benefits

Decreased renal accumulation of aminoglycoside reflects defective receptor-mediated endocytosis in cystic fibrosis and Dent's disease

The clinical use of aminoglycoside (AG) antibiotics is limited by their renal toxicity, which is caused by drug accumulation in proximal tubule (PT) cells. Clinical studies reported that renal clearance of AG is enhanced in cystic fibrosis (CF) patients, which might reflect the role of CFTR in PT cell endocytosis. In order to assess the role of chloride transporters on the renal handling of AG, we investigated gentamicin uptake and renal accumulation in mice lacking functional CFTR (Cftr ∆F/∆F) or knock-out for the Cl−/H+ exchanger ClC-5 (Clcn5 Y/− ). The latter represent a paradigm of PT dysfunction and defective receptor-mediated endocytosis. As compared with controls, Cftr ∆F/∆F and Clcn5 Y/− mice showed a 15% to 85% decrease in gentamicin accumulation in the kidney, respectively, in absence of renal failure. Studies on primary cultures of Cftr ∆F/∆F and Clcn5 Y/− mouse PT cells confirmed the reduction in gentamicin uptake, although colocalization with endosomes and lysosomes was maintained. Quantification of endocytosis in PT cells revealed that gentamicin, similar to albumin, preferentially binds to megalin. The functional loss of ClC-5 or CFTR was reflected by a decrease of the endocytic uptake of gentamicin, with a more pronounced effect in cells lacking ClC-5. These results support the concept that CFTR, as well as ClC-5, plays a relevant role in PT cell endocytosis. They also demonstrate that the functional loss of these two chloride transporters is associated with impaired uptake of AG in PT cells, reflected by a decreased renal accumulation of the dru

Análise da Transformação da Paisagem na Vertente Oeste do Alto Curso do ARROIO GUABIROBA/ SANANDUVA-RS, DE 1996 A 2015

Estudar a transformação da paisagem a partir da análise temporal e espacial significa compreender as relações físicas e sociais de um determinado espaço geográfico, pois, através desta análise é possível identificar os fatores que influenciaram e influenciam na sua modificação. A pesquisa busca analisar as transformações ocorridas na paisagem por meio do uso e cobertura do solo da vertente oeste no alto e médio curso do arroio Guabiroba no município de Sananduva- RS, entre os anos de 1996 e 2015. Sendo assim, buscou-se contextualizar histórico e espacialmente a transformação da paisagem a partir do povoamento do local, apontando os principais usos do solo ao longo do tempo histórico em escala local. Como produto da análise constatou-se que a vegetação exótica tomou espaço de algumas áreas que em 1996 eram mata nativa e agricultura. No contexto geral na área de estudo a agricultura predomina incentivado pela modernização agrícola. Desse modo, a modernização agrícola acelerou o processo de transformação da paisagem e promoveu a redução da vegetação natural na área de estudo

Azithromycin fails to reduce increased expression of neutrophil-related cytokines in primary-cultured epithelial cells from cystic fibrosis mice

AbstractBackgroundBeneficial effects of azithromycin in cystic fibrosis (CF) have been reported, however, its mechanism of action remains unclear. The present study aimed at investigating the effect of azithromycin on CF airway epithelial cells.MethodsPrimary cultures of purified tracheal epithelial cells from F508del and normal homozygous mice were established. Responses to lipopolysaccharide from Pseudomonas aeruginosa (LPS, 0.1 µg/ml) on mRNA expression of neutrophil-related chemokines, pro- and anti-inflammatory cytokines were investigated in the presence or the absence of azithromycin (1 µg/ml).ResultsCF airway epithelial cells showed upregulation of MIP-2 and KC responses to LPS, and azithromycin failed to downregulate these responses. In contrast, in CF cells, azithromycin increased KC and TNF-α expression under non-stimulated and LPS-stimulated conditions, respectively. In non-CF cells, the macrolide potentiated the LPS response on MIP-2 and on IL-10.ConclusionsAirway epithelial cells contribute to the dysregulated immune processes in CF. Azithromycin rather stimulates cytokine expression in CF airway epithelial cells

Entre caminhos e descaminhos, a objetivação do ser mais como um pilar para a pesquisa em Ensino de Ciências e Biologia

O Núcleo de Estudos em Ensino de Genética, Biologia e Ciências (NUEG) tem se dedicado, ao longo das duas últimas décadas, às atividades de Pesquisa, Ensino e Extensão, realizando estudos e investigações baseados em aspectos sociais, históricos e culturais que envolvem a produção e disseminação de conhecimentos sobre formação de professores e ensino e aprendizagem de Ciências e Biologia. A partir dos pressupostos freireanos da dialogicidade, problematização e humanização, o grupo vem construindo uma perspectiva teórico-metodológica problematizadora, cuja orientação coletiva baseia-se em cinco questões crítico-reflexivas. Compreendemos que essa práxis investigativa está comprometida com a formação permanente de educadores/pesquisadores

Sweat Proteomics in Cystic Fibrosis: Discovering Companion Biomarkers for Precision Medicine and Therapeutic Development

peer reviewedIn clinical routine, the diagnosis of cystic fibrosis (CF) is still challenging regardless of international consensus on diagnosis guidelines and tests. For decades, the classical Gibson and Cooke test measuring sweat chloride concentration has been a keystone, yet, it may provide normal or equivocal results. As of now, despite the combination of sweat testing, CFTR genotyping, and CFTR functional testing, a small fraction (1–2%) of inconclusive diagnoses are reported and justifies the search for new CF biomarkers. More importantly, in the context of precision medicine, with a view to early diagnosis, better prognosis, appropriate clinical follow-up, and new therapeutic development, discovering companion biomarkers of CF severity and phenotypic rescue are of utmost interest. To date, previous sweat proteomic studies have already documented disease-specific variations of sweat proteins (e.g., in schizophrenia and tuberculosis). In the current study, sweat samples from 28 healthy control subjects and 14 patients with CF were analyzed by nanoUHPLC-Q-Orbitrap-based shotgun proteomics, to look for CF-associated changes in sweat protein composition and abundance. A total of 1057 proteins were identified and quantified at an individual level, by a shotgun label-free approach. Notwithstanding similar proteome composition, enrichment, and functional annotations, control and CF samples featured distinct quantitative proteome profiles significantly correlated with CF, accounting for the respective inter-individual variabilities of control and CF sweat. All in all: (i) 402 sweat proteins were differentially abundant between controls and patients with CF, (ii) 68 proteins varied in abundance between F508del homozygous patients and patients with another genotype, (iii) 71 proteins were differentially abundant according to the pancreatic function, and iv) 54 proteins changed in abundance depending on the lung function. The functional annotation of pathophysiological biomarkers highlighted eccrine gland cell perturbations in: (i) protein biosynthesis and trafficking, (ii) CFTR proteostasis and membrane stability, and (iii) cell-cell adherence, membrane integrity, and cytoskeleton crosstalk. Cytoskeleton-related biomarkers were of utmost interest because of the consistency between variations observed here in CF sweat and variations previously documented in other CF tissues. From a clinical stance, nine candidate biomarkers of CF diagnosis (CUTA, ARG1, EZR, AGA, FLNA, MAN1A1, MIA3, LFNG, SIAE) and seven candidate biomarkers of CF severity (ARG1, GPT, MDH2, EML4 (F508del homozygous), MGAT1 (pancreatic insufficiency), IGJ, TOLLIP (lung function impairment)) were deemed suitable for further verification.MucoSweatOmic

Dysregulated Proinflammatory and Fibrogenic Phenotype of Fibroblasts in Cystic Fibrosis

Morbi-mortality in cystic fibrosis (CF) is mainly related to chronic lung infection and inflammation, uncontrolled tissue rearrangements and fibrosis, and yet the underlying mechanisms remain largely unknown. We evaluated inflammatory and fibrosis responses to bleomycin in F508del homozygous and wild-type mice, and phenotype of fibroblasts explanted from mouse lungs and skin. The effect of vardenafil, a cGMP-specific phosphodiesterase type 5 inhibitor, was tested in vivo and in culture. Responses of proinflammatory and fibrotic markers to bleomycin were enhanced in lungs and skin of CF mice and were prevented by treatment with vardenafil. Purified lung and skin fibroblasts from CF mice proliferated and differentiated into myofibroblasts more prominently and displayed higher sensitivity to growth factors than those recovered from wild-type littermates. Under inflammatory stimulation, mRNA and protein expression of proinflammatory mediators were higher in CF than in wild-type fibroblasts, in which CFTR expression reached similar levels to those observed in other non-epithelial cells, such as macrophages. Increased proinflammatory responses in CF fibroblasts were reduced by half with submicromolar concentrations of vardenafil. Proinflammatory and fibrogenic functions of fibroblasts are upregulated in CF and are reduced by vardenafil. This study provides compelling new support for targeting cGMP signaling pathway in CF pharmacotherapy

SERVIÇOS E PRODUTOS DO SIBi/USP: descrição dos processos essenciais, gerenciais e de apoio

O Modelo de Gestão do Sistema Integrado de Bibliote\ud cas da Universidade de São Paulo\ud (SIBi/USP) incorpora conceitos e ferramentas de ger\ud enciamento que permeiam as\ud organizações modernas para garantir melhores índice\ud s de desempenho sistêmico e prestar\ud serviços com qualidade e eficiência aos usuários. C\ud omo parte do Modelo está a\ud identificação e a descrição detalhada dos processos\ud de trabalho do Sistema, assim como o\ud estabelecimento de alguns indicadores de desempenho\ud . A partir do\ud trabalho inicial, foi\ud elaborado um levantamento complementar para identif\ud icação dos processos que\ud abrangessem o conjunto do Sistema de forma ampla. O\ud s dados foram dispostos em\ud planilhas para melhor visualização, especialmente q\ud uanto aos macro processos, processos,\ud sub processos e atividades. Os processos foram sepa\ud rados em essenciais, gerenciais e de\ud apoio, além de elencadas as atividades pertinentes\ud a cada um deles, bem como as\ud instruções técnicas e fluxos de trabalho. Foram est\ud abelecidos alguns indicadores, tendo por\ud referência os da IFLA já estudados por outro Grupo\ud de Trabalho. Daquele estudo quatro\ud indicadores foram testados e validados pelo SIBi/US\ud P por meio de aplicação piloto em\ud algumas das Bibliotecas do Sistema e outros foram d\ud efinidos no estudo atual. Com isso foi\ud possível mapear os processos e as atividades desenv\ud olvidas pelo conjunto de Bibliotecas\ud sendo que cada Biblioteca, em função de sua especia\ud lidade e especificidade pode adequar\ud o seu mapeamento. A definição de um núcleo básico d\ud e indicadores objetiva viabilizar a\ud concretização da missão e dos objetivos em consonân\ud cia com a política do SIBI/USPThe Management Model of the Integrated Library Syst\ud em, University of São Paulo\ud (SIBi/USP) incorporates concepts and management too\ud ls that permeate modern\ud organizations to ensure better systemic performance\ud indexes and provide services\ud with quality and efficiency to users. Is part of th\ud e model, the identification and the\ud detailed description of the essential processes, ma\ud nagement and support, as well as\ud the establishment of some performance indicators. F\ud rom the initial work, we designed\ud a complementary survey to identify the processes co\ud vering the whole of the System.\ud Data were arranged in sheets for easy viewing, espe\ud cially with regard to macro \ud processes, processes, sub processes and activities.\ud The processes were divided into\ud essential, managerial and support. At that time, we\ud re listed the activities under each\ud processes and their technical instructions and work\ud flows. It were established some\ud indicators, using model IFLA already studied by oth\ud er Working Group. In this study\ud four indicators have been tested and validated by S\ud IBi/USP through pilot application\ud in some of the Library System and others were defin\ud ed in the current study. It was\ud possible to map the processes and activities undert\ud aken by all Libraries of SIBi/USP,\ud and each of them, depending on their specialty and\ud specificity can do its own\ud mapping. The definition of a core of indicators can\ud enable the achievement of the\ud mission and objectives in line with the policy of S\ud IBI/US

Entre caminhos e descaminhos, a objetivação do ser mais como um pilar para a pesquisa em Ensino de Ciências e Biologia

O Núcleo de Estudos em Ensino de Genética, Biologia e Ciências (NUEG) tem se dedicado, ao longo das duas últimas décadas, às atividades de Pesquisa, Ensino e Extensão, realizando estudos e investigações baseados em aspectos sociais, históricos e culturais que envolvem a produção e disseminação de conhecimentos sobre formação de professores e ensino e aprendizagem de Ciências e Biologia. A partir dos pressupostos freireanos da dialogicidade, problematização e humanização, o grupo vem construindo uma perspectiva teórico-metodológica problematizadora, cuja orientação coletiva baseia-se em cinco questões crítico-reflexivas. Compreendemos que essa práxis investigativa está comprometida com a formação permanente de educadores/pesquisadores

HLA-G expression and regulation during Pseudomonas aeruginosa infection in cystic fibrosis patients

Deregulated immune response fails to control biofilm-forming bacteria, as Pseudomonas aeruginosa, in the lungs of cystic fibrosis (CF) patients. HLA-G is an immune-modulatory molecule involved in respiratory diseases and infections