1,720,984 research outputs found
Molecular basis of cystic fibrosis: from bench to bedside
No full textCystic fibrosis (CF), is an autosomal recessive disease affecting different organs. The lung disease, characterized by recurrent and chronic bacterial infection and inflammation since infancy, is the main cause of morbidity and precocious mortality of these individuals. The innovative therapies directed to repair the defective CF gene should account for the presence of more than 200 disease-causing mutations of the CF transmembrane conductance regulator (CFTR) gene. The review will recall the different experimental approaches in discovering CFTR protein targeted molecules, such as the high throughput screening on chemical libraries to discover correctors and potentiators of CFTR protein, dual-acting compounds, read-through molecules, splicing defects repairing tools, CFTR "amplifiers"
Revealing the microRNA-transcription factors network in cystic fibrosis:from microRNA therapeutics to precision medicine (CF-miRNA-THER) FFC 7/2018
No full textThe project by Gambari et al. proposes to identify miRNAs that regulate (i) CFTR, (ii) transcription factors that regulate CFTR, and (iii) factors that enhance CFTR cell surface expression. Next, they will design and synthesize PNAs and PNA masks to inhibit these miRNAs and their interactions with specific mRNAs with important roles in CF, respectively; the effect of these PNA molecules and combinations thereof on gene expression and the secretome will be assessed. MiRNAs in various CF body fluids will be quantified. Overall the project aims to develop new therapeutics to enhance CFTR expression/function that may be used as adjunct therapies for people with CF
Cystic fibrosis carriers have higher neonatal immunoreactive trypsinogen values than non-carriers
No full textFollowing cystic fibrosis (CF) neonatal screening implementation, a high frequency of heterozygotes has been reported among neonates with elevated immunoreactive trypsinogen (IRT) and normal sweat chloride levels. We studied the relationship between normal IRT values and CF heterozygosity: 10,000 neonates were screened for CF by IRT measurement and tested for 40 CF mutations; the 294 carriers detected were coupled with newborns negative to the same genetic testing, and the two groups' IRT levels compared. Heterozygotes had higher IRT levels than their controls (mean 35.32 vs. 27.58 microg/L, P<0.001). Even within normal trypsinogen range, the probability of being a CF carrier increases with neonatal IRT concentration
Induction of IL-6 gene expression in a CF bronchial epithelial cell line by Pseudomonas aeruginosa is dependent on transcription factors belonging to the Sp1 superfamily
No full textCystic fibrosis (CF) is a genetic disease characterized by chronic bacterial lung infection, most commonly sustained by Pseudomonas aeruginosa. Upon infection, elevated concentrations of pro-inflammatory cytokines (i.e. IL-6 and IL1beta) and chemokines (i.e. IL-8 and GROgamma) are found in the bronchoalveolar fluid of CF patients. We report in this paper that: (a) IL-8, IL-6, IL-1beta, ICAM-1, and GRO-gamma genes are upregulated following infection of CF bronchial epithelial IB3-1 cells with P. aeruginosa; (b) Sp1 transcription factor activity is induced following infection of the cystic fibrosis IB3-1 and CuFi-1 cell lines; (c) inhibition of Sp1 activity using transcription factor decoy molecules leads to inhibition of the expression of IL-6 gene. From the theoretical point of view, our results demonstrate that Sp1 transcription factor activity is induced following infection of CF cells with P. aeruginosa, and that this effect is important in the activation of IL-6 gene transcription. From the practical point of view, our data sustain the potential use of decoy molecules targeting the transcription factor Sp1 to control a relevant molecule involved in the inflammatory process associated with the cystic fibrosis airway pathology
Silencing of genes coding for transcription factors: biological effects of decoy oligonucleotides on cystic fibrosis bronchial epithelial cells.
No full textChronic pulmonary inflammation in the lungs of patients affected by cystic fibrosis is characterized by massive intra-bronchial recruitment of neutrophils, initiated and sustained upon interaction of pathogens (including Pseudomonas aeruginosa) with surface bronchial cells, associated with release of chemokines, including interleukins IL-8 and IL-6. This process leads to progressive tissue damage. In order to develop novel therapeutic strategies altering this process, the transcription factor (TF) decoy approach was applied, based on the intracellular delivery of double stranded oligodeoxynucleotides (ODNs) mimicking the binding sites of transcription factors and causing inhibition of the binding of TF-related proteins to regulatory sequences present in the promoter of specific genes. Since the promoters IL-8 and IL-6 contain consensus sequences for NF-kB and Sp1, IB3-1 cells were transfected with double stranded TF “decoy” ODNs targeting NF-kB and Sp1. Inhibition of Sp1 activity using transcription factor decoy molecules leads to inhibition of the expression of IL-6 gene; in addition, NF-kB decoy ODNs produced inhibition of transcription of IL-8. In conclusion, these results suggest that intracellular delivery of “decoy” molecules aimed to compete with the NF-kB and Sp1 transcription factors is a promising strategy to obtain selective inhibition of expression of IL-8 and IL-6 genes stimulated in epithelial cells of the respiratory tract upon surface interaction with bacteria
Transcription Factor Oligodeoxynucleotides to NF-{kappa}B Inhibit Transcription of IL-8 in Bronchial Cells.
No full textChronic pulmonary inflammation in patients affected by cystic fibrosis (CF) is characterized by massive bronchial infiltrates of neutrophils, which is sustained by the interaction of pathogens (e.g. Pseudomonas aeruginosa) with surface bronchial cells. To explore new treatment options focused on the reduction of neutrophil chemotaxis, we applied the transcription factor (TF) decoy approach, based on the intracellular delivery of double stranded oligodeoxynucleotides (ODNs) causing inhibition of the binding of TF-related proteins to the different consensus sequences in the promoter of specific genes. In CF bronchial IB3-1 cells, P.aeruginosa induced transcription of the neutrophil chemokines IL-8 and GRO-gamma of the adhesion molecule ICAM-1, and of the cytokines IL-1beta and IL-6. Since consensus sequences for the TF, NF-kappaB, is contained in the promoters of all these genes, IB3-1, CuFi-1, Beas-2B and CaLu-3 cells were transfected with double stranded TF "decoy" ODNs mimicking different NF-kappaB consensus sequences. IL-8 NF-kappaB decoy ODN partially inhibited the P.aeruginosa-dependent transcription of IL-8, GRO-gamma and IL-6, whereas decoy ODNs to both HIV-1 LTR and Igk produced a strong, 80 to 85% inhibition of transcription of IL-8, without reducing that of GRO-gamma, ICAM-1, IL-1beta and IL-6. In conclusion, intracellular delivery of "decoy" molecules aimed to compete with the TF, NF-kappaB, is a promising strategy to obtain inhibition of of IL-8 gene transcription
Expression of miR-93 and IL-8 during Pseudomonas aeruginosa infection of the human bronchial cystic fibrosis IB3-1 cells
No full textIn this study we have first analyzed by microarray and RT-qPCR the miR-profile of cystic fibrosis IB3-1 cells infected with Pseudomonas aeruginosa, demonstrating miR-93 as highly expressed and reduced during infection, in association with increased expression of the IL-8 gene. Sequence analysis shows that the 3’UTR region of the IL-8 mRNA is a potential target of miR-93 and highly conserved throughout molecular evolution. The involvement of miR-93 in IL-8 gene expression was demonstrated by the finding that down-modulation of miR-93 following P.aeruginosa infection was counteracted by pre-miR-93 transfection. In addition, in uninfected IB3-1 cells treated with antagomiR-93, IL-8 was up-regulated. Since miR-93 is located in the 13th intron of the gene of MCM7 (minichromosome maintenance protein 7), the expression of MCM7 gene was studied in parallel with miR-93 and the miR-93 target IL-8 mRNA. After P.aeruginosa infection, a decrease of both MCM7 and miR-93 transcripts occurs in parallel with increase of IL-8 production. The decrease of MCM7/miR-93 transcription was associated with decrease of nuclear recruitment of MYC and E2F-1 transcription factors, involved in the regulation of the MCM7 gene promoter. To our knowledge, this is the first observation of a possible link between micro RNA expression and IL-8 induction in cystic fibrosis cells infected by Pseudomonas aeruginosa. The results reported in the present paper, therefore, suggest that in addition to transcriptional NF-kB dependent up-regulation of IL-8 gene expression, IL-8 protein secretion might depend from interactions of the IL-8 mRNA with inhibitory micro RNAs, including miR-93
Antibacterial and anti-inflammatory activity of a temporin B peptide analogue on an in vitro model of cystic fibrosis
No full textNatural peptides with antimicrobial properties are deeply investigated as tools to fight bacteria resistant to common antibiotics. Small peptides, as those belonging to the temporin family, are very attractive because their activity can easily be tuned after small modification to their primary sequence. Structure-activity studies previously reported by us allowed the identification of one peptide, analogue of temporin B, TB_KKG6A, showing, unlike temporin B, antimicrobial activity against both Gram-positive and Gram-negative bacteria. In this paper, we investigated the antimicrobial and anti-inflammatory activity of the peptide TB_KKG6A against Pseudomonas aeruginosa. Interestingly, we found that the peptide exhibits antimicrobial activity at low concentrations, being able to downregulate the pro-inflammatory chemokines and cytokines interleukin (IL)-8, IL-1β, IL-6 and tumor necrosis factor-α produced downstream infected human bronchial epithelial cells. Experiments were carried out also with temporin B, which was found to show pro-inflammatory activity. Details on the interaction between TB_KKG6A and the P. aeruginosa LPS were obtained by circular dichroism and fluorescence studies
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
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