Extra-respiratory comorbidities and transplantation in the French cystic fibrosis registry.

Expert Rev Respir Med. 2019 Jul 03;:1-4

Authors: Mainbourg S, Durieu I, Dehillotte C, Reynaud Q

Abstract
Background: To better understand the potential role of lung transplantation (LT) in accelerating or slowing down extrarespiratory cystic fibrosis (CF) complications, we compare the respective prevalence of extrapulmonary CF comorbidities in transplanted and nontransplanted adult CF patients. Methods: About 20% of the adult CF patients included in the French CF registry have undergone transplantation. We analyzed all adults included in the French CF registry in 2014 and compared them according to their transplant status, irrespective of the year of transplantation, to describe the prevalence of CF-related diabetes (CFRD), end-stage renal disease and osteoporosis (Chi-squared and Fisher's exact tests). Results: A total of 3339 adult CF patients were included in the French CF Registry and 673 (20.2%) were transplanted. Prevalence of CFRD was significantly higher in transplanted patients compared to nontransplanted patients irrespective of the age group; the prevalence of end-stage renal disease and osteoporosis was also significantly higher in transplanted patients. Conclusion: These results demonstrate the high prevalence of extrapulmonary comorbidities in transplanted CF adult patients particularly CFRD. We highlight the need for specific surveillance and prevention for transplanted CF patients who were already treated for extra espiratory comorbidities before lung transplantation, due to the potential aggravation of their comorbidities after transplantation.

PMID: 31267788 [PubMed - as supplied by publisher]

Mild cystic fibrosis in carriers of two nonsense mutations - a case of genetic compensation response?

J Cyst Fibros. 2019 Jun 29;:

Authors: Tümmler B

PMID: 31266708 [PubMed - as supplied by publisher]

Exercising our options: comparing effects of exercise and positive expiratory pressure on mucociliary clearance.

Eur Respir J. 2019 Apr;53(4):

Authors: Donaldson SH

PMID: 31000670 [PubMed - indexed for MEDLINE]

Proteostasis Takes Center Stage in Pulmonary Fibrosis.

Am J Respir Cell Mol Biol. 2019 Jun;60(6):605-606

Authors: Budinger GRS

PMID: 30849235 [PubMed - indexed for MEDLINE]

Long-Term CFTR Modulators and Changes in Hemoglobin.

Ann Am Thorac Soc. 2019 Mar;16(3):305-306

Authors: Middleton PG

PMID: 30821493 [PubMed - indexed for MEDLINE]

Ion the Prize: Defining the Complexities of Airway Epithelial Cell Ion Transport Functions.

Am J Respir Cell Mol Biol. 2019 Jun;60(6):618-620

Authors: Nguyen JP, Hirota JA

PMID: 30768913 [PubMed - indexed for MEDLINE]

CFTR is not a gluten lover either.

EMBO J. 2019 Jan 15;38(2):

Authors: Vachel L, Muallem S

PMID: 30573671 [PubMed - indexed for MEDLINE]

The Emerging Role of Neutrophil Extracellular Traps in Respiratory Disease.

Chest. 2019 Jun 29;:

Authors: Twaddell SH, Baines KJ, Grainge C, Gibson PG

Abstract
Neutrophil extracellular traps (NETs) are extrusions of intracellular DNA and attached granular material which enable bacterial killing. NETs are increasingly recognised for their role in the pathogenesis of respiratory disease. NETs are composed of a complex mix of intracellular derived material which neutrophils organise within the cytoplasm then expel in a non-directed manner in the vicinity of invading organisms. Combined, these trap and destroy multiple genera of microbes including bacteria, fungi, viruses and protozoans, limiting infection especially where phagocytosis is not possible. Initially NET formation was thought to be a terminal event for neutrophils, however it is now apparent that some neutrophils survive this process becoming anuclear and may drive ongoing tissue damage. NETs are now known to be directly cytotoxic to lung epithelium and endothelium and their excessive production is seen in pneumonia and acute lung injury as well as several chronic diseases, including COPD, asthma and cystic fibrosis. NETs also appear to play a role in both tumour defence and dissemination depending on the local microenvironment and the specific tumour subtype. It is becoming increasingly apparent that NET formation can be a positive or negative influence on multiple respiratory pathologies and that simply globally reducing or increasing NET formation is unlikely to be a therapeutic success. Rather, as our understanding grows, it is likely that targeted NET up or downregulation along with destruction or protection of already formed NETs may be become an additional point of intervention for respiratory physicians.

PMID: 31265835 [PubMed - as supplied by publisher]

[Intestinal microbiota, nutrients and probiotics viewed from the «gut-lung» axis].

Vopr Pitan. 2019;88(3):13-22

Authors: Zolnikova OY, Ivaschkin KV, Bueverova EL, Ivaschkin VT

Abstract
Disturbance of the bronchopulmonary system are among the most common and socially significant diseases, so, the prevention and treatment of these disorders are the priority tasks of practical health care. Being based on the accumulated literature data on the interaction of the intestinal microflora and respiratory tract, the role of symbiotic bacteria of the intestinal biotope has been discussed in the respiratory diseases' pathogenesis. The aim of the work was to analyze the results of experimental and clinical studies confirming the effect of intestinal microflora on the development and progression of respiratory diseases. The analysis of the available data on the risk reducing of occurrence, duration and severity of symptoms of bronchial asthma when taking probiotics, both in childhood and in the adult population, has been carried out. The effectiveness of the probiotic microorganisms' intake for the treatment of chronic obstructive pulmonary disease, pneumonia, viral infection, cystic fibrosis, and lung cancer has been analyzed. The main possible molecular mechanisms of the symbiotic bacteria prevention of the bronchopulmonary diseases development have been discussed in the article. Conclusion. The probiotics usage in the complex treatment of bronchopulmonary diseases demonstrates encouraging results. Its potential may be useful in the treatment of various lung diseases. However, a number of questions have been related to the individual selection of specific strains, the dosage and duration of use to achieve sustained remission for a patient.

PMID: 31265771 [PubMed - in process]

Normal Calcium-Activated Anion Secretion in a Mouse Selectively Lacking TMEM16A in Intestinal Epithelium.

Front Physiol. 2019;10:694

Authors: Vega G, Guequén A, Johansson MEV, Arike L, Martínez-Abad B, Scudieri P, Pedemonte N, Millar-Büchner P, Philp AR, Galietta LJ, Hansson GC, Flores CA

Abstract
Calcium-activated anion secretion is expected to ameliorate cystic fibrosis, a genetic disease that carries an anion secretory defect in exocrine tissues. Human patients and animal models of the disease that present a mild intestinal phenotype have been postulated to bear a compensatory calcium-activated anion secretion in the intestine. TMEM16A is calcium-activated anion channel whose presence in the intestinal epithelium is contradictory. We aim to test the functional expression of TMEM16A using animal models with Cftr and/or Tmem16a intestinal silencing. Expression of TMEM16A was studied in a wild type and intestinal Tmem16a knockout mice by mRNA-seq, mass-spectrometry, q-PCR, Western blotting and immunolocalization. Calcium-activated anion secretion was recorded in the ileum and proximal colon of these animals including intestinal Cftr knockout and double mutants with dual Tmem16a and Cftr intestinal ablation. Mucus homeostasis was studied by immune-analysis of Mucin-2 (Muc2) and survival curves were recorded. Tmem16a transcript was found in intestine. Nevertheless, protein was barely detected in colon samples. Electrophysiological measurements demonstrated that the intestinal deletion of Tmem16a did not change calcium-activated anion secretion induced by carbachol or ATP in ileum and proximal colon. Muc2 architecture was not altered by Tmem16a silencing as was observed when Cftr was deleted from mouse intestine. Tmem16a silencing neither affected animal survival nor modified the lethality observed in the intestinal Cftr-null mouse. Our results demonstrate that TMEM16A function in the murine intestine is not related to electrogenic calcium-activated anion transport and does not affect mucus homeostasis and survival of animals.

PMID: 31263421 [PubMed]

Azithromycin Polarizes Macrophages to an M2 Phenotype via Inhibition of the STAT1 and NF-κB Signaling Pathways.

J Immunol. 2019 Jul 01;:

Authors: Haydar D, Cory TJ, Birket SE, Murphy BS, Pennypacker KR, Sinai AP, Feola DJ

Abstract
Azithromycin is effective at controlling exaggerated inflammation and slowing the long-term decline of lung function in patients with cystic fibrosis. We previously demonstrated that the drug shifts macrophage polarization toward an alternative, anti-inflammatory phenotype. In this study we investigated the immunomodulatory mechanism of azithromycin through its alteration of signaling via the NF-κB and STAT1 pathways. J774 murine macrophages were plated, polarized (with IFN-γ, IL-4/-13, or with azithromycin plus IFN-γ) and stimulated with LPS. The effect of azithromycin on NF-κB and STAT1 signaling mediators was assessed by Western blot, homogeneous time-resolved fluorescence assay, nuclear translocation assay, and immunofluorescence. The drug's effect on gene and protein expression of arginase was evaluated as a marker of alternative macrophage activation. Azithromycin blocked NF-κB activation by decreasing p65 nuclear translocation, although blunting the degradation of IκBα was due, at least in part, to a decrease in IKKβ kinase activity. A direct correlation was observed between increasing azithromycin concentrations and increased IKKβ protein expression. Moreover, incubation with the IKKβ inhibitor IKK16 decreased arginase expression and activity in azithromycin-treated cells but not in cells treated with IL-4 and IL-13. Importantly, azithromycin treatment also decreased STAT1 phosphorylation in a concentration-dependent manner, an effect that was reversed with IKK16 treatment. We conclude that azithromycin anti-inflammatory mechanisms involve inhibition of the STAT1 and NF-κB signaling pathways through the drug's effect on p65 nuclear translocation and IKKβ.

PMID: 31263039 [PubMed - as supplied by publisher]

Dissociation of systemic and mucosal autoimmunity in cystic fibrosis.

J Cyst Fibros. 2019 Jun 28;:

Authors: Theprungsirikul J, Skopelja-Gardner S, Meagher RE, Clancy JP, Zemanick ET, Ashare A, Rigby WFC

Abstract
BACKGROUND: Pseudomonas aeruginosa accounts for ~80% of cystic fibrosis (CF) airway infection. It shows a remarkable correlation with presence of autoantibody to bactericidal/permeability-increasing protein (BPI), which is not understood. In this study, we sought to better understand the characteristics of systemic and mucosal autoimmunity and their relation to humoral immunity to P. aeruginosa.
METHODS: Antibody titers and isotypes to BPI and P. aeruginosa were characterized in sera and bronchoalveolar lavage (BAL) of adult and pediatric CF patients (n = 131), by ELISA and/or immunoblot.
RESULTS: Serum BPI autoantibodies were common (~43%) in adult while rare (≪5%) in pediatric (≤18 yrs) CF patients. Serum BPI IgG autoantibodies were of high avidity and strongly correlated with anti-P. aeruginosa IgG responses. A parallel relationship was observed with IgA, but not IgG, responses in adult and pediatric CF patient in the BAL. Thus, BAL IgA anti-BPI antibodies were independent of age and correlated with the presence of BPI cleavage in BAL.
CONCLUSIONS: IgG and IgA autoreactivity to BPI in CF patients was demonstrated in serum and BAL, respectively, and correlated with the isotype of the antibody response to P. aeruginosa. The co-occurrence of anti-BPI and anti-P. aeruginosa IgA in the BAL, but not serum, of pediatric CF patients suggests that BPI tolerance is broken in the P. aeruginosa-infected airway and that serologic IgG autoantibodies are later induced, potentially through a separate pathway. The relationship between P. aeruginosa, BPI cleavage, and IgA autoantibodies in the BAL suggests a role for cryptic epitope generation in the breaking of tolerance.

PMID: 31262645 [PubMed - as supplied by publisher]

CFTR regulates B cell activation and lymphoid follicle development.

Respir Res. 2019 Jul 01;20(1):133

Authors: Polverino F, Lu B, Quintero JR, Vargas SO, Patel AS, Owen CA, Gerard NP, Gerard C, Cernadas M

Abstract
BACKGROUND: Cystic fibrosis (CF) is an inherited disorder caused by mutations in the CF transmembrane conductance regulator (CFTR) gene that promotes persistent lung infection and inflammation and progressive loss of lung function. Patients with CF have increased lung lymphoid follicles (LFs) and B cell-activating factor of tumor necrosis factor family (BAFF) that regulates B cell survival and maturation. A direct role for CFTR in B cell activation and disease pathogenesis in CF remains unclear.
METHODS: The number of LFs, BAFF+, TLR4+ and proliferation marker Ki67+ B cells in lung explants or resections from subjects with CF and normal controls was quantified by immunostaining. The role of CFTR in B cell activation and LF development was then examined in two independent cohorts of uninfected CFTR-deficient mice (Cftr -/-) and wild type controls. The number of lung LFs, B cells and BAFF+, CXCR4+, immunoglobulin G+ B cells was examined by immunostaining. Lung and splenocyte B cell activation marker and major histocompatibility complex class II (MHC class II) expression was quantified by flow cytometry. Inflammatory cytokine levels were measured in supernatants from isolated B cells from Cftr -/- and wild type mice stimulated in vitro with Pseudomonas aeruginosa lipopolysaccharide (LPS).
RESULTS: There was a significant increase in well-formed LFs in subjects with CF compared to normal controls. Increased B cell activation and proliferation was observed in lung LFs from CF subjects as was quantified by a significant increase in B cell BAFF, TLR4 and Ki67 expression. Uninfected Cftr -/- mice had increased lung LFs and BAFF+ and CXCR4+ B cells compared to wild type controls. Lung B cells isolated from uninfected Cftr -/- mice demonstrated increased MHC class II expression. In vitro, isolated B cells from Cftr -/- mice produced increased IL-6 when stimulated with LPS compared to wild type controls.
CONCLUSIONS: These data support a direct role for CFTR in B cell activation, proliferation and inflammatory cytokine production that promotes lung LF follicle development in cystic fibrosis.

PMID: 31262295 [PubMed - in process]

The Prevalence of Cigarette and E-cigarette Smoking Among Students in Central and Eastern Europe-Results of the YUPESS Study.

Int J Environ Res Public Health. 2019 Jun 28;16(13):

Authors: Brożek GM, Jankowski M, Lawson JA, Shpakou A, Poznański M, Zielonka TM, Klimatckaia L, Loginovich Y, Rachel M, Gereová J, Golonko J, Naumau I, Kornicki K, Pepłowska P, Kovalevskiy V, Raskiliene A, Bielewicz K, Krištúfková Z, Mróz R, Majek P, Lubanski J, Kaleta D, Pinkas J, Zejda JE

Abstract
Electronic cigarettes (e-cigarettes) are an alternative to traditional tobacco cigarette smoking. The aim of this study was to assess the prevalence of cigarette smoking and e-cigarette use among university students from Central and Eastern Europe and to investigate personal characteristics associated with cigarette and e-cigarette smoking. A questionnaire-based cross-sectional survey was performed between 2017-2018 among university students in five European countries: Belarus, Lithuania, Poland, Russia, and Slovakia. The questionnaire included 46 questions related to the frequency and habits of traditional cigarettes and e-cigarettes use. Completed questionnaires were obtained from 14,352 students (8800 medical; aged 20.9 ± 2.4 years) with an overall response rate of 72.2%. Two-thirds of the respondents had smoked a traditional tobacco cigarette and 43.7% had used an e-cigarette. Overall current smoking status included 12.3% traditional cigarette smokers, 1.1% e-cigarette users, and 1.8% were dual users with the remainder being non-smokers. Smoking status differed between the research centres (p < 0.001). Females were less likely to try either cigarettes (OR = 0.83) or e-cigarettes (OR = 0.62) and were less likely to be current cigarette (OR = 0.64), e-cigarette (OR = 0.34), or dual users (OR = 0.33) than males. Perception of e-cigarettes significantly differed between smokers and non-smokers (p < 0.001). Among university students, cigarettes are more popular than e-cigarettes.

PMID: 31261697 [PubMed - in process]

Characterisation of the Major Extracellular Proteases of Stenotrophomoas maltophilia and Their Effects on Pulmonary Antiproteases.

Pathogens. 2019 Jun 28;8(3):

Authors: Molloy K, Smith SG, Cagney G, Dillon ET, Greene CM, McElvaney NG

Abstract
Stenotrophomonas maltophilia is an emerging global opportunistic pathogen that has been appearing with increasing prevalence in cystic fibrosis (CF). A secreted protease from S. maltophilia has been reported as its chief potential virulence factor. Here, using the reference clinical strain S. maltophilia K279a, the major secreted proteases were identified. Protein biochemistry and mass spectrometry were carried out on K279a culture supernatant. The effect of K279a culture supernatant on cleavage and anti-neutrophil elastase activity of the three majors pulmonary antiproteases was quantified. A deletion mutant of S. maltophilia lacking expression of a protease was constructed. The serine proteases StmPR1, StmPR2 and StmPR3, in addition to chitinase A and an outer membrane esterase were identified in culture supernatants. Protease activity was incompletely abrogated in a K279a-ΔStmPR1: Erm mutant. Wild type K279a culture supernatant degraded alpha-1 antitrypsin (AAT), secretory leucoprotease inhibitor (SLPI) and elafin, important components of the lung's innate immune defences. Meanwhile SLPI and elafin, but not AAT, retained their ability to inhibit neutrophil elastase. StmPR3 together with StmPR1 and StmPR2, is likely to contribute to protease-mediated innate immune dysfunction in CF.

PMID: 31261656 [PubMed]

SpartaABC: a web server to simulate sequences with indel parameters inferred using an approximate Bayesian computation algorithm.

Nucleic Acids Res. 2017 07 03;45(W1):W453-W457

Authors: Ashkenazy H, Levy Karin E, Mertens Z, Cartwright RA, Pupko T

Abstract
Many analyses for the detection of biological phenomena rely on a multiple sequence alignment as input. The results of such analyses are often further studied through parametric bootstrap procedures, using sequence simulators. One of the problems with conducting such simulation studies is that users currently have no means to decide which insertion and deletion (indel) parameters to choose, so that the resulting sequences mimic biological data. Here, we present SpartaABC, a web server that aims to solve this issue. SpartaABC implements an approximate-Bayesian-computation rejection algorithm to infer indel parameters from sequence data. It does so by extracting summary statistics from the input. It then performs numerous sequence simulations under randomly sampled indel parameters. By computing a distance between the summary statistics extracted from the input and each simulation, SpartaABC retains only parameters behind simulations close to the real data. As output, SpartaABC provides point estimates and approximate posterior distributions of the indel parameters. In addition, SpartaABC allows simulating sequences with the inferred indel parameters. To this end, the sequence simulators, Dawg 2.0 and INDELible were integrated. Using SpartaABC we demonstrate the differences in indel dynamics among three protein-coding genes across mammalian orthologs. SpartaABC is freely available for use at http://spartaabc.tau.ac.il/webserver.

PMID: 28460062 [PubMed - indexed for MEDLINE]

Update on the current modalities used to screen high risk youth for prediabetes and/or type 2 diabetes mellitus.

Ann Pediatr Endocrinol Metab. 2019 Jun;24(2):71-77

Authors: Brar PC

Abstract
The modalities currently employed to screen for type 2 diabetes mellitus (T2DM)/prediabetes are HbA1c, fasting plasma glucose (FPG), and 2-hour plasma glucose (PG) during an oral glucose tolerance test (OGTT). The purpose of this review is to highlight the positive qualities and pitfalls of these diagnostic modalities and reflect on the most reasonable and effective approach to screen high risk youth. Given its inherent preanalytical advantages, glycated hemoglobin (HbA1c) continues to be the preferred diagnostic modality used by pediatricians to screen high risk youth. However, when the three aforementioned tests are performed in youths of different races/ethnicities, discrepant results for T2DM/prediabetes are observed. The prevalence rates for T2DM vary from 0.53% in Chinese youth (including youth of all body mass indexes) to 18.3% in high-risk, overweight, obese Korean youth. Moreover, the FPG is abnormal (&gt;100 less than &lt;126 mg/dL) in 15% of Korean youth versus 8.7% of Chinese youth. The prevalence rates for prediabetes are 1.49% in Chinese youth versus 21% in Emirati youth (HbA1c, 5.7%-6.4%). The coefficient of agreement, k, between these screening tests for T2DM are fair, 0.45-0.5 across all youth. However, using HbA1c as a comparator, the agreement is weak with FPG (k=0.18 in German youth versus k=0.396 in Korean youth). The American Diabetes Association (ADA) Standards of Medical Care Guidelines define "high risk youth" who need to be tested for T2DM and/or prediabetes. OGTT and HbA1c do not always detect T2DM in similar individuals. HbA1c may not be an ideal test for screening Hispanic and African American youth. FPG and OGTT are suitable screening tests for youth of ethnic minorities and those with cystic fibrosis or hemoglobinopathies. Performing all three tests either together or sequentially may be the only way to encompass all youth who have aberrations in different aspects of glucose homeostasis.

PMID: 31261470 [PubMed]

Gastrointestinal, Pancreatic, and Hepatic Manifestations of Cystic Fibrosis in the Newborn.

Neoreviews. 2019 Jan;20(1):e12-e24

Authors: Galante G, Freeman AJ

Abstract
Gastrointestinal, pancreatic, and hepatic signs and symptoms represent the most common presentation of early disease among patients with cystic fibrosis and may be the initial indication of disease. Regardless of whether cystic fibrosis is diagnosed early by newborn screening or later by clinical course, the impact of gastrointestinal, pancreatic, and hepatic manifestations on early life is nearly ubiquitous. Conditions strongly linked with cystic fibrosis, such as meconium ileus and pancreatic insufficiency, must be recognized and treated early to optimize both short- and long-term care. Similarly, less specific conditions such as reflux, poor weight gain, and cholestasis are frequently encountered in infants with cystic fibrosis. In this population, these conditions may present unique challenges in which early interventions may have significant influence on both short- and long-term morbidity and mortality outcomes.

PMID: 31261070 [PubMed - in process]

Real-Time, Semi-Automated Fluorescent Measurement of the Airway Surface Liquid pH of Primary Human Airway Epithelial Cells.

J Vis Exp. 2019 Jun 13;(148):

Authors: Saint-Criq V, Haq IJ, Gardner AI, Garnett JP, Ward C, Brodlie M, Gray MA

Abstract
In recent years, the importance of mucosal surface pH in the airways has been highlighted by its ability to regulate airway surface liquid (ASL) hydration, mucus viscosity and activity of antimicrobial peptides, key parameters involved in innate defense of the lungs. This is of primary relevance in the field of chronic respiratory diseases such as cystic fibrosis (CF) where these parameters are dysregulated. While different groups have studied ASL pH both in vivo and in vitro, their methods report a relatively wide range of ASL pH values and even contradictory findings regarding any pH differences between non-CF and CF cells. Furthermore, their protocols do not always provide enough details in order to ensure reproducibility, most are low throughput and require expensive equipment or specialized knowledge to implement, making them difficult to establish in most labs. Here we describe a semi-automated fluorescent plate reader assay that enables the real-time measurement of ASL pH under thin film conditions that more closely resemble the in vivo situation. This technique allows for stable measurements for many hours from multiple airway cultures simultaneously and, importantly, dynamic changes in ASL pH in response to agonists and inhibitors can be monitored. To achieve this, the ASL of fully differentiated primary human airway epithelial cells (hAECs) are stained overnight with a pH-sensitive dye in order to allow for the reabsorption of the excess fluid to ensure thin film conditions. After fluorescence is monitored in the presence or absence of agonists, pH calibration is performed in situ to correct for volume and dye concentration. The method described provides the required controls to make stable and reproducible ASL pH measurements, which ultimately could be used as a drug discovery platform for personalized medicine, as well as adapted to other epithelial tissues and experimental conditions, such as inflammatory and/or host-pathogen models.

PMID: 31259916 [PubMed - in process]

Enabling Synthesis of ABBV-2222, A CFTR Corrector for the Treatment of Cystic Fibrosis.

Org Lett. 2019 Jul 01;:

Authors: Greszler SN, Shelat B, Voight EA

Abstract
An enabling preclinical synthetic route to cystic fibrosis candidate ABBV-2222 is described. Two stereoselective steps provide access to an aminochroman intermediate with excellent control, and a late-stage demethylation/difluoromethylation sequence provides efficient access to the target molecule.

PMID: 31259557 [PubMed - as supplied by publisher]