Epiregulin (EREG) is upregulated through an IL-1β autocrine loop in Caco-2 Epithelial Cells with Reduced CFTR Function.

J Cell Biochem. 2017 Nov 01;:

Authors: Massip-Copiz M, Clauzure M, Valdivieso ÁG, Santa-Coloma TA

Abstract
CFTR is a cAMP-regulated chloride channel, whose mutations produce cystic fibrosis. The impairment of CFTR activity increases the intracellular Cl(-) concentration, which in turn produces an increased interleukin-1β (IL-1β) secretion. The secreted IL-1β then induces an autocrine positive feedback loop, further stimulating IL-1β priming and secretion. Since IL-1β can transactivate the epidermal growth factor receptor (EGFR), we study here the levels of expression for different EGFR ligands in Caco-2/pRS26 cells (expressing shRNA against CFTR resulting in a reduced CFTR expression and activity). The epiregulin (EREG), amphiregulin (AREG) and heparin binding EGF like growth factor (HBEGF) mRNAs, were found overexpressed in Caco-2/pRS26 cells. The EREG mRNA had the highest differential expression and was further characterized. In agreement with its mRNA levels, Western blots (WB) showed increased EREG levels in CFTR-impaired cells. In addition, EREG mRNA and protein levels were stimulated by incubation with exogenous IL-1β and inhibited by the Interleukin 1 receptor type I (IL1R1) antagonist IL1RN, suggesting that the overexpression of EREG is a consequence of the autocrine IL-1β loop previously described for these cells. In addition, the JNK inhibitor SP600125, and the EGFR inhibitors AG1478 and PD168393, also had an inhibitory effect on EREG expression, suggesting that EGFR, activated in Caco-2/pRS26 cells, is involved in the observed EREG upregulation. In conclusion, in Caco-2 CFTR-shRNA cells, the EGFR ligand EREG is overexpressed due to an active IL-1β autocrine loop that indirectly activates EGFR, constituting new signaling effectors for the CFTR signaling pathway, downstream of CFTR, Cl(-) and IL-1β. This article is protected by copyright. All rights reserved.

PMID: 29091309 [PubMed - as supplied by publisher]

Physical exercise training for cystic fibrosis.

Cochrane Database Syst Rev. 2017 Nov 01;11:CD002768

Authors: Radtke T, Nevitt SJ, Hebestreit H, Kriemler S

Abstract
BACKGROUND: Physical exercise training may form an important part of regular care for people with cystic fibrosis. This is an update of a previously published review.
OBJECTIVES: To assess the effects of physical exercise training on exercise capacity by peak oxygen consumption, pulmonary function by forced expiratory volume in one second, health-related quality of life and further important patient-relevant outcomes in people with cystic fibrosis.
SEARCH METHODS: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group Trials Register which comprises references identified from comprehensive electronic database searches and handsearches of relevant journals and abstract books of conference proceedings.Date of the most recent search: 04 May 2017.We searched ongoing trials registers (clinicaltrials.gov and the WHO ICTRP). Date of most recent search: 10 August 2017.
SELECTION CRITERIA: All randomised and quasi-randomised controlled clinical trials comparing exercise training of any type and a minimum duration of two weeks with conventional care (no training) in people with cystic fibrosis.
DATA COLLECTION AND ANALYSIS: Two authors independently selected studies for inclusion, assessed methodological quality and extracted data. The quality of the evidence was assessed using the GRADE system.
MAIN RESULTS: Of the 83 studies identified, 15 studies which included 487 participants, met the inclusion criteria. The numbers in each study ranged from nine up to 72 participants; two studies were in adults, seven were in children and adolescents and six studies included all age ranges. Four studies of hospitalised participants lasted less than one month and 11 studies were outpatient-based, lasting between two months and three years. The studies included participants with a wide range of disease severity and employed differing levels of supervision with a mixture of types of training. There was also wide variation in the quality of the included studies.This systematic review shows very low- to low-quality evidence from both short- and long-term studies that in people with cystic fibrosis aerobic or anaerobic physical exercise training (or a combination of both) has a positive effect on aerobic exercise capacity, pulmonary function and health-related quality of life. No study reported on mortality; two studies reported on adverse events (moderate-quality evidence); one of each study reported on pulmonary exacerbations (low-quality evidence) and diabetic control (very low-quality evidence). Although improvements were not consistent between studies and ranged from no effects to clearly positive effects, the most consistent effects of the heterogeneous exercise training modalities and durations were found for maximal aerobic exercise capacity (in four out of seven studies) with unclear effects on forced expiratory volume in one second (in two out of 11 studies) and health-related quality of life (in two out of seven studies).
AUTHORS' CONCLUSIONS: Evidence about the efficacy of physical exercise training in cystic fibrosis from 15 small studies with low to moderate methodological quality is limited. Exercise training is already part of regular outpatient care offered to most people with cystic fibrosis, and since there is some evidence for beneficial effects on aerobic fitness and no negative side effects exist, there is no reason to actively discourage this. The benefits from including physical exercise training in an individual's regular care may be influenced by the type and duration of the training programme. High-quality randomised controlled trials are needed to comprehensively assess the benefits of exercise programmes in people with cystic fibrosis and the relative benefits of the addition of aerobic versus anaerobic versus a combination of both types of physical exercise training to the care of people with cystic fibrosis.

PMID: 29090734 [PubMed - as supplied by publisher]

Activation of TPA-response element present in human Lemur Tyrosine Kinase 2 (lmtk2) gene increases its expression.

Biochem Biophys Rep. 2017 Dec;12:140-150

Authors: Dey I, Bradbury NA

Abstract
Regulatory elements present in the promoter of a gene drive the expression of the gene in response to various stimuli. Lemur Tyrosine Kinase 2 (LMTK2) is a membrane-anchored Serine/Threonine kinase involved in endosomal protein trafficking and androgen signaling amongst other processes. Previous studies have shown this protein to be of therapeutic importance in cystic fibrosis and prostate cancer. However, nothing is known about the endogenous expression of this protein and its regulation. In this study, we analyzed the gene encoding human LMTK2, to look for possible regulatory elements that could affect its expression. Interestingly, the human lmtk2 gene contains a consensus TPA (12- O-Tetradecanoylphorbol-13-acetate)-responsive element (TRE) in the region preceding its start codon. The element with the sequence TGAGTCA modulates LMTK2 expression in response to treatment with TPA, a synthetic Protein Kinase C (PKC) activator. It serves as the binding site for c-Fos, a member of the Activator Protein -1 (AP-1) transcription factor complex, which is transactivated by PKC. We observed that TPA, at low concentrations, increases the promoter activity of LMTK2, which leads to a subsequent increase in the mRNA transcript and protein levels. This modulation occurs through binding of the AP-1 transcription factor complex to the lmtk2 promoter. Thus, our current study has established LMTK2 as a TPA-responsive element-containing gene, which is upregulated downstream of PKC activation. Considering the involvement of LMTK2 in intracellular processes as well as pathological conditions, our findings demonstrate a way to modulate intracellular LMTK2 levels pharmacologically for potentially therapeutic purposes.

PMID: 29090275 [PubMed]

Neonatal Gastrointestinal Perforations: the 10-Year Experience of a Reference Hospital.

Indian J Surg. 2017 Oct;79(5):431-436

Authors: Saraç M, Bakal Ü, Aydın M, Tartar T, Orman A, Taşkın E, Canpolat Ş, Kazez A

Abstract
The aim of this study was to present our experiences with, as well as the factors that affect, the treatment and outcome of patients with neonatal gastrointestinal perforations (GIPs). Thirty-eight newborn cases that were operated on for GIP in our hospital's tertiary newborn intensive care unit between January 2005 and December 2015 were retrospectively evaluated. The patients were divided into the two following groups: group 1, perforations related to necrotizing enterocolitis (NEC), and group 2, non-NEC perforations. In total, 38 patients (16 males, 22 females) participated in this study. The perforations were related to NEC in 12 patients (group 1; 31.6 %), and the other 26 patients (group 2; 68.4 %) were classified as non-NEC perforation cases. The incidence of neonatal GIP was 0.53 % in all newborn patients, while the incidence of perforation in NEC cases was 20 %. Of all patients, 25 (65.7 %) were premature. Non-NEC pathologies were the most common cause of GIP (68.4 %) and included stomach perforation related to a nasogastric catheter (n = 5), volvulus (n = 4), intestinal atresia (n = 3), esophageal atresia and tracheoesophageal fistula (n = 2), cystic fibrosis (n = 2), Hirschprung's disease (n = 2), appendicitis (n = 2), congenital stomach anterior wall weakness (n = 1), duplication cyst (n = 1), invagination (n = 1), incarcerated inguinal hernia (n = 1), and idiopathic causes (n = 2). Primary surgical repair was performed in all cases without a conservative approach. The mortality rate related to GIP in newborn cases was 47.3 %. While the mortality rate in group 1 was 66.6 %, it was statistically insignificantly lower in group 2 (38.4 %) (p > 0.05). In group 1, the mortality rate of those with intestinal and colorectal perforations was 45.6 and 20 %, respectively (p > 0.05). Non-NEC pathologies are the most frequent causes of GIP in newborns, and primary surgical repair is the primary treatment choice for neonatal GIP. However, GIP remains one of the most significant causes of mortality in newborns. While the prognosis for neonatal colon perforation is good, that for stomach and jejunoileal perforations is worse.

PMID: 29089704 [PubMed - in process]

Differential Effects of Angelicin Analogues on NF-κB Activity and IL-8 Gene Expression in Cystic Fibrosis IB3-1 Cells.

Mediators Inflamm. 2017;2017:2389487

Authors: Lampronti I, Manzione MG, Sacchetti G, Ferrari D, Spisani S, Bezzerri V, Finotti A, Borgatti M, Dechecchi MC, Miolo G, Marzaro G, Cabrini G, Gambari R, Chilin A

Abstract
The angelicin analogue 4,6,4'-trimethylangelicin (TMA) was recently reported as a strong inhibitor of nuclear factor-κB (NF-κB) activity and of the expression of the interleukin-8 (IL-8) gene in bronchial epithelial cells in which the inflammatory response has been challenged with P. aeruginosa, the most common bacterium found in the airways of patients affected by cystic fibrosis (CF). These findings encouraged us to analyze new synthetic analogues of TMA in order to evaluate their biological activities on human bronchial epithelial CF IB3-1 cells and to find more potent anti-NF-κB agents exhibiting only minor antiproliferative effects. Analogues able to inhibit NF-κB/DNA interaction at lower concentration than TMA were found and selected to investigate their biological activity on IB3-1 cells induced with TNF-α. In this biological system, NF-κB-mediated IL-8 gene expression was investigated. Some analogues showed similar activity to the lead compound TMA. Other analogues displayed higher activities; in particular, the most interesting compounds showing relevant anti-inflammatory effects were found to cause 56-83% reduction of IL-8 mRNA expression at low concentrations (1-10 μM), without changes in cell proliferation pattern, demonstrating their potential interest for a possible development of anti-inflammatory therapy of cystic fibrosis.

PMID: 29089668 [PubMed - in process]

Evolved Aztreonam Resistance Is Multifactorial and Can Produce Hypervirulence in Pseudomonas aeruginosa.

MBio. 2017 Oct 31;8(5):

Authors: Jorth P, McLean K, Ratjen A, Secor PR, Bautista GE, Ravishankar S, Rezayat A, Garudathri J, Harrison JJ, Harwood RA, Penewit K, Waalkes A, Singh PK, Salipante SJ

Abstract
While much attention has been focused on acquired antibiotic resistance genes, chromosomal mutations may be most important in chronic infections where isolated, persistently infecting lineages experience repeated antibiotic exposure. Here, we used experimental evolution and whole-genome sequencing to investigate chromosomally encoded mutations causing aztreonam resistance in Pseudomonas aeruginosa and characterized the secondary consequences of resistance development. We identified 19 recurrently mutated genes associated with aztreonam resistance. The most frequently observed mutations affected negative transcriptional regulators of the mexAB-oprM efflux system and the target of aztreonam, ftsI While individual mutations conferred modest resistance gains, high-level resistance (1,024 µg/ml) was achieved through the accumulation of multiple variants. Despite being largely stable when strains were passaged in the absence of antibiotics, aztreonam resistance was associated with decreased in vitro growth rates, indicating an associated fitness cost. In some instances, evolved aztreonam-resistant strains exhibited increased resistance to structurally unrelated antipseudomonal antibiotics. Surprisingly, strains carrying evolved mutations which affected negative regulators of mexAB-oprM (mexR and nalD) demonstrated enhanced virulence in a murine pneumonia infection model. Mutations in these genes, and other genes that we associated with aztreonam resistance, were common in P. aeruginosa isolates from chronically infected patients with cystic fibrosis. These findings illuminate mechanisms of P. aeruginosa aztreonam resistance and raise the possibility that antibiotic treatment could inadvertently select for hypervirulence phenotypes.IMPORTANCE Inhaled aztreonam is a relatively new antibiotic which is being increasingly used to treat cystic fibrosis patients with Pseudomonas aeruginosa airway infections. As for all antimicrobial agents, bacteria can evolve resistance that decreases the effectiveness of the drug; however, the mechanisms and consequences of aztreonam resistance are incompletely understood. Here, using experimental evolution, we have cataloged spontaneous mutations conferring aztreonam resistance and have explored their effects. We found that a diverse collection of genes contributes to aztreonam resistance, each with a small but cumulative effect. Surprisingly, we found that selection for aztreonam resistance mutations could confer increased resistance to other antibiotics and promote hypervirulence in a mouse infection model. Our study reveals inherent mechanisms of aztreonam resistance and indicates that aztreonam exposure can have unintended secondary effects.

PMID: 29089424 [PubMed - in process]

The challenge of measuring intra-individual change in fatigue during cancer treatment.

Qual Life Res. 2017 Feb;26(2):259-271

Authors: Moinpour CM, Donaldson GW, Davis KM, Potosky AL, Jensen RE, Gralow JR, Back AL, Hwang JJ, Yoon J, Bernard DL, Loeffler DR, Rothrock NE, Hays RD, Reeve BB, Smith AW, Hahn EA, Cella D

Abstract
PURPOSE: To evaluate how well three different patient-reported outcomes (PROs) measure individual change.
METHODS: Two hundred and fourteen patients (from two sites) initiating first or new chemotherapy for any stage of breast or gastrointestinal cancer participated. The 13-item FACIT Fatigue scale, a 7-item PROMIS(®) Fatigue Short Form (PROMIS 7a), and the PROMIS(®) Fatigue computer adaptive test (CAT) were administered monthly online for 6 months. Reliability of measured change was defined, under a population mixed effects model, as the ratio of estimated systematic variance in rate of change to the estimated total variance of measured individual differences in rate of change. Precision of individual measured change, the standard error of measurement of change, was given by the square root of the rate-of-change sampling variance. Linear and quadratic models were examined up to 3 and up to 6 months.
RESULTS: A linear model for measured change showed the following by 6 and 3 months, respectively: PROMIS CAT (0.363 and 0.342); PROMIS SF (0.408 and 0.533); FACIT (0.459 and 0.473). Quadratic models offered no noteworthy improvement over linear models. Both reliability and precision results demonstrate the need to improve the measurement of intra-individual change.
CONCLUSIONS: These results illustrate the challenge of reliably measuring individual change in fatigue with a level of confidence required for intervention. Optimizing clinically useful measurement of intra-individual differences over time continues to pose a challenge for PROs.

PMID: 27469506 [PubMed - indexed for MEDLINE]

High attainment of optimal nutritional and growth status observed among Greek pediatric cystic fibrosis patients: results from the GreeCF study.

J Pediatr Endocrinol Metab. 2017 Oct 26;30(11):1169-1176

Authors: Poulimeneas D, Petrocheilou A, Grammatikopoulou MG, Kaditis AG, Loukou I, Doudounakis SE, Laggas D, Vassilakou T

Abstract
BACKGROUND: Pediatric cystic fibrosis (CF) patients suffer high rates of undernutrition, subject to several parameters. We aimed to assess growth and nutritional status of Greek children and adolescents with CF.
METHODS: Eighty-four patients (35 boys) formed the sample. Anthropometrics and FEV1 were measured, growth and weight status were assessed. Body mass index (BMI), arm circumference (MUAC), fat (MUAFA) and muscle (MUAMA) were calculated.
RESULTS: In the total sample, 6.0% of the patients were underweight, 4.8% stunted, 8.3% wasted and 17.9% in nutritional failure, whereas 59.5% attained the ideal BMI for CF. FEV1 positively associated with BMI (B=0.03, p≤0.003), weight (B=0.03, p≤0.003) and MUAMA z-scores (B=0.04, p≤0.005). Meconium ileus negatively associated with FEV1 (B=-14.17, p≤0.003) and stature (B=-0.65, p≤0.043). Pancreatic insufficiency negatively influenced MUAC and MUAFA z-scores (p≤0.05 for both).
CONCLUSIONS: The examined CF patients appear to be thriving. Unlike published research, the participants' sex, gene mutation and acquisition of pathogens did not affect growth.

PMID: 29087958 [PubMed - in process]

Pulmonary ventilation imaging in asthma and cystic fibrosis using oxygen-enhanced 3D radial ultrashort echo time MRI.

J Magn Reson Imaging. 2017 Oct 31;:

Authors: Zha W, Kruger SJ, Johnson KM, Cadman RV, Bell LC, Liu F, Hahn AD, Evans MD, Nagle SK, Fain SB

Abstract
BACKGROUND: A previous study demonstrated the feasibility of using 3D radial ultrashort echo time (UTE) oxygen-enhanced MRI (UTE OE-MRI) for functional imaging of healthy human lungs. The repeatability of quantitative measures from UTE OE-MRI needs to be established prior to its application in clinical research.
PURPOSE: To evaluate repeatability of obstructive patterns in asthma and cystic fibrosis (CF) with UTE OE-MRI with isotropic spatial resolution and full chest coverage.
STUDY TYPE: Volunteer and patient repeatability.
POPULATION: Eighteen human subjects (five asthma, six CF, and seven normal subjects).
FIELD STRENGTH/SEQUENCE: Respiratory-gated free-breathing 3D radial UTE (80 μs) sequence at 1.5T.
ASSESSMENT: Two 3D radial UTE volumes were acquired sequentially under normoxic and hyperoxic conditions. A subset of subjects underwent repeat acquisitions on either the same day or ≤15 days apart. Asthma and CF subjects also underwent spirometry. A workflow including deformable registration and retrospective lung density correction was used to compute 3D isotropic percent signal enhancement (PSE) maps. Median PSE (MPSE) and ventilation defect percent (VDP) of the lung were measured from the PSE map.
STATISTICAL TESTS: The relations between MPSE, VDP, and spirometric measures were assessed using Spearman correlations. The test-retest repeatability was evaluated using Bland-Altman analysis and intraclass correlation coefficients (ICC).
RESULTS: Ventilation measures in normal subjects (MPSE = 8.0%, VDP = 3.3%) were significantly different from those in asthma (MPSE = 6.0%, P = 0.042; VDP = 21.7%, P = 0.018) and CF group (MPSE = 4.5%, P = 0.0006; VDP = 27.2%, P = 0.002). MPSE correlated significantly with forced expiratory lung volume in 1 second percent predicted (ρ = 0.72, P = 0.017). The ICC of the test-retest VDP and MPSE were both ≥0.90. In all subject groups, an anterior/posterior gradient was observed with higher MPSE and lower VDP in the posterior compared to anterior regions (P ≤ 0.0021 for all comparisons).
DATA CONCLUSION: 3D radial UTE OE-MRI supports quantitative differentiation of diseased vs. healthy lungs using either whole lung VDP or MPSE with excellent test-retest repeatability.
LEVEL OF EVIDENCE: 2 Technical Efficacy: Stage 1 J. Magn. Reson. Imaging 2017.

PMID: 29086454 [PubMed - as supplied by publisher]

Prevalence of Fecal Incontinence in Adults with Cystic Fibrosis.

Dig Dis Sci. 2017 Oct 30;:

Authors: Benezech A, Desmazes-Dufeu N, Baumstarck K, Bouvier M, Coltey B, Reynaud-Gaubert M, Vitton V

Abstract
BACKGROUND: Patients with cystic fibrosis (CF) are deemed at risk of developing urinary incontinence (UI) due to repeated coughing and other factors causing increased pressure on the pelvic floor. Fecal incontinence (FI) is probably derived from the same mechanism, but only very few data are available on its frequency.
AIMS: The aim of this study was to determine the prevalence of FI in an adult population with CF.
METHODS: This retrospective study was conducted from January 2012 to June 2014. Patients were recruited from Marseille referral center for adult CF. They were asked to fill in a self-completed anonymous questionnaire for symptom assessment of UI and FI. Clinical data and a detailed history of CF were also recorded.
RESULTS: A total of 155 out of 190 patients (92 females) of mean age 30.5 ± 11 years completed the survey. Seventy-three patients (47%) were lung transplanted. Forty patients (25.8%) reported FI with a mean St Mark's score of 4.9 ± 2. Thirty-five patients (22.6%) reported UI. Eighteen patients (11.6%) reported both FI and UI. FI was significantly more frequent in older patients (34.27 vs. 29.54 years, p = 0.03) and in patients with associated UI (p = 0.001). No relationship was found between respiratory, bacterial, nutritional status, transplantation, pancreatic status, practice of physiotherapy, delivery history, and FI.
CONCLUSIONS: The high prevalence of FI in CF and its negative impacts need to integrate this symptom in the overall treatment of this pathology. The systematic early detection of FI may allow its rapid management and limit their consequences.

PMID: 29086331 [PubMed - as supplied by publisher]

Safety of intravenous tobramycin in combination with a variety of anti-pseudomonal antibiotics in children with cystic fibrosis.

SAGE Open Med. 2017;5:2050312117736694

Authors: Deschamp AR, Pettit RS, Donaldson JA, Slaven JE, Davis SD

Abstract
OBJECTIVES: Previous studies have examined renal safety of once daily intravenous tobramycin in individuals with cystic fibrosis. This has been mainly in combination with ceftazidime in an adolescent or adult population. In this report, we describe our institutional experience of once daily intravenous tobramycin in combination with a variety of second anti-pseudomonal antibiotics in children with cystic fibrosis.
METHODS: We present a retrospective review including children with cystic fibrosis, who were admitted for a pulmonary exacerbation from January 2009 to December 2011, and treated using intravenous tobramycin. A literature review of once daily intravenous aminoglycoside dosing in cystic fibrosis was performed to compare our results to existing literature.
RESULTS: A total of 35 subjects were divided into once daily dosing (n = 20) versus multiple daily dosing (n = 15) groups. Mean age was 11.3 years (± 5.7) for the once daily dosing group and 13.1 years (± 4.4) for the multiple daily dosing group (p = 0.34). All subjects had normal baseline serum creatinine at admission (once daily dosing 0.49 ± 0.14 mg/dL vs multiple daily dosing 0.62 ± 0.23 mg/dL, p = 0.07). All subjects received intravenous tobramycin, and most received piperacillin-tazobactam as their second anti-pseudomonal antibiotic (once daily dosing 45% and multiple daily dosing 40%). There was no significant change in serum creatinine in either group during antibiotic treatment (once daily dosing 0.08 ± 0.12 mg/dL vs. multiple daily dosing 0.06 ± 0.10 mg/dL, p = 0.43). All subjects had significant improvement in lung function following intravenous antibiotic therapy.
CONCLUSION: We show that both once daily dosing and multiple daily dosing of intravenous tobramycin in combination with a variety of second anti-pseudomonal antibiotics were safe in terms of nephrotoxicity in children with cystic fibrosis. These findings are important given existing literature mainly examines once daily tobramycin in combination with ceftazidime, a cephalosporin, and the majority of our patients were on tobramycin with piperacillin-tazobactam, an extended spectrum penicillin plus beta-lactam. This contributes new information not previously examined in a pediatric cystic fibrosis population.

PMID: 29085640 [PubMed]

Klotho Inhibits Interleukin-8 Secretion from Cystic Fibrosis Airway Epithelia.

Sci Rep. 2017 Oct 30;7(1):14388

Authors: Krick S, Baumlin N, Aller SP, Aguiar C, Grabner A, Sailland J, Mendes E, Schmid A, Qi L, David NV, Geraghty P, King G, Birket SE, Rowe SM, Faul C, Salathe M

Abstract
Chronic inflammation is a hallmark of cystic fibrosis (CF) and associated with increased production of transforming growth factor (TGF) β and interleukin (IL)-8. α-klotho (KL), a transmembrane or soluble protein, functions as a co-receptor for Fibroblast Growth Factor (FGF) 23, a known pro-inflammatory, prognostic marker in chronic kidney disease. KL is downregulated in airways from COPD patients. We hypothesized that both KL and FGF23 signaling modulate TGF β-induced IL-8 secretion in CF bronchial epithelia. Thus, FGF23 and soluble KL levels were measured in plasma from 48 CF patients and in primary CF bronchial epithelial cells (CF-HBEC). CF patients showed increased FGF23 plasma levels, but KL levels were not different. In CF-HBEC, TGF-β increased KL secretion and upregulated FGF receptor (FGFR) 1. Despite increases in KL, TGF-β also increased IL-8 secretion via activation of FGFR1 and Smad 3 signaling. However, KL excess via overexpression or supplementation decreased IL-8 secretion by inhibiting Smad 3 phosphorylation. Here, we identify a novel signaling pathway contributing to IL-8 secretion in the CF bronchial epithelium with KL functioning as an endocrine and local anti-inflammatory mediator that antagonizes pro-inflammatory actions of FGF23 and TGF-β.

PMID: 29085059 [PubMed - in process]

CF Airway Microbiome: Overturning the Old, Opening the Way for the New.

J Bacteriol. 2017 Oct 30;:

Authors: O'Toole GA

Abstract
The genetic disease cystic fibrosis (CF) is associated with chronic airway infections that are a proximal cause of death in many patients with this affliction. Classic microbiology studies focusing on canonical pathogens resulted in the development of a common set of views regarding the nature of the airway infections associated with this disease, and these ideas have influenced everything from the way infections are treated to how clinical trials for new CF-targeted antibiotics are designed and the focus of CF-related research topics. Recent culture-independent studies have prompted us to rethink, and in some cases discard, some of these long-held views. In this piece, I argue that an updated view of the complicated chronic infections associated with CF, thanks in large part to culture-independent studies of sputum and bronchoalveolar lavage samples, should be leveraged to develop new strategies to treat these recalcitrant infections.

PMID: 29084859 [PubMed - as supplied by publisher]

Functional classification of gill ionocytes and spatiotemporal changes in their distribution after transfer from seawater to fresh water in Japanese seabass.

J Exp Biol. 2017 Oct 30;:

Authors: Inokuchi M, Nakamura M, Miyanishi H, Hiroi J, Kaneko T

Abstract
Spatiotemporal changes in branchial ionocyte distribution were investigated following transfer from seawater (SW) to fresh water (FW) in Japanese seabass. The mRNA expression levels of cystic fibrosis transmembrane conductance regulator (CFTR) and Na(+)/K(+)/2Cl(-) cotransporter 1a (NKCC1a) in the gills rapidly decreased after transfer to FW, whereas Na(+)/H(+) exchanger 3 (NHE3) and Na(+)/Cl(-) cotransporter 2 (NCC2) expressions were upregulated following the transfer. By quadruple-color whole-mount immunofluorescence staining with anti-Na(+)/K(+)-ATPase, anti-NHE3, anti-CFTR and T4 (anti-NKCC1a/NCC2) antibodies, we classified ionocytes into one SW-type and two FW-types; NHE3 cell and NCC2 cell. Time-course observation after transfer revealed an intermediate type between SW-type and FW-type NHE3 ionocytes, suggesting functional plasticity of ionocytes. Finally, on the basis of the ionocyte classification of Japanese seabass, we observed the location of ionocyte subtypes on frozen sections of the gill filaments stained by triple-color immunofluorescence staining. Our observation indicated that SW-type ionocytes transformed into FW-type NHE3 ionocytes and at the same time shifted their distribution from filaments to lamellae. On the other hand, FW-specific NCC2 ionocytes appeared mainly in the filaments. Taken together, these findings indicated that ionocytes originated from undifferentiated cells in the filaments and expanded their distribution to the lamellae during FW acclimation.

PMID: 29084852 [PubMed - as supplied by publisher]

Use of Calgary and microfluidic BioFlux systems to test the activity of fosfomycin and tobramycin alone and in combination against cystic fibrosis Pseudomonas aeruginosa biofilms.

Antimicrob Agents Chemother. 2017 Oct 30;:

Authors: Díez-Aguilar M, Morisini MI, Köksal E, Oliver A, Ekkelenkamp M, Cantón R

Abstract
Pseudomonas aeruginosa is a major cause of morbidity and mortality in chronically infected cystic fibrosis patients. Novel in vitro biofilm models, which reliably predict therapeutic success of antimicrobial therapies, should be implemented. The activity of fosfomycin, tobramycin and fosfomycin-tobramycin combination was tested against 6 susceptible P. aeruginosa strains isolated from respiratory samples of cystic fibrosis patients by using two in vitro biofilm models: a closed system (Calgary device) and an open model based on microfluidics (BioFlux). All but one of the isolates formed biofilm. The fosfomycin and tobramycin minimal biofilm inhibitory concentrations (MBIC) were 1,024->1,024 μg/ml and 8-32 μg/ml, respectively. According to fractional inhibitory concentration analysis, the combination behaved synergistically in all the isolates except in the P. aeruginosa ATCC 27853 strain.The dynamic formation of the biofilm was also studied with the BioFlux system and the MIC and MBIC of each antibiotic were tested. For the combination, the lowest tobramycin concentration that was synergistic with fosfomycin was used. The captured images were analyzed measuring the intensity of colored pixels, which is proportional to the biofilm biomass. A statistically significant difference was found when comparing the intensity of the inoculum with the intensity in the microchannel where the MBIC of tobramycin or fosfomycin or their combination was used (p<0.01) but not when applying the MIC (p>0.01).Fosfomycin-tobramycin demonstrated to be synergistic against cystic fibrosis P. aeruginosa strains in biofilm models, both when testing with the Calgary and the microfluidic BioFlux systems. These results support the clinical use of this combination.

PMID: 29084746 [PubMed - as supplied by publisher]

The Escherichia coli rhaSR-PrhaBAD Inducible Promoter System Allows Tightly Controlled Gene Expression over a Wide Range in Pseudomonas aeruginosa.

Appl Environ Microbiol. 2016 Nov 15;82(22):6715-6727

Authors: Meisner J, Goldberg JB

Abstract
The araC-ParaBAD inducible promoter system is tightly controlled and allows gene expression to be modulated over a wide range in Escherichia coli, which has led to its widespread use in other bacteria. Although anecdotal evidence suggests that araC-ParaBAD is leaky in Pseudomonas aeruginosa, neither a thorough analysis of this inducible promoter system in P. aeruginosa nor a concerted effort to identify alternatives with improved functionality has been reported. Here, we evaluated the functionality of the araC-ParaBAD system in P. aeruginosa Using transcriptional fusions to a lacZ reporter gene, we determined that the noninduced expression from araC-ParaBAD is high and cannot be reduced by carbon catabolite repression as it can in E. coli Modulating translational initiation by altering ribosome-binding site strength reduced the noninduced activity but also decreased the maximal induced activity and narrowed the induction range. Integrating the inducible promoter system into the posttranscriptional regulatory network that controls catabolite repression in P. aeruginosa significantly decreased the noninduced activity and increased the induction range. In addition to these improvements in the functionality of the araC-ParaBAD system, we found that the lacI(q)-Ptac and rhaSR-PrhaBAD inducible promoter systems had significantly lower noninduced expression and were inducible over a broader range than araC-ParaBAD We demonstrated that noninduced expression from the araC-ParaBAD system supported the function of genes involved in antibiotic resistance and tryptophan biosynthesis in P. aeruginosa, problems that were avoided with rhaSR-PrhaBAD. rhaSR-PrhaBAD is tightly controlled, allows gene expression over a wide range, and represents a significant improvement over araC-ParaBAD in P. aeruginosa IMPORTANCE: We report the shortcomings of the commonly used Escherichia coli araC-ParaBAD inducible promoter system in Pseudomonas aeruginosa, successfully reengineered it to improve its functionality, and show that the E. coli rhaSR-PrhaBAD system is tightly controlled and allows inducible gene expression over a wide range in P. aeruginosa.

PMID: 27613678 [PubMed - indexed for MEDLINE]

Whole-lung torsion complicating double lung transplantation: CT features.

Diagn Interv Imaging. 2016 Sep;97(9):927-8

Authors: David A, Liberge R, Corne F, Frampas E

PMID: 27421675 [PubMed - indexed for MEDLINE]

Pulmonary function disparities exist and persist in Hispanic patients with cystic fibrosis: A longitudinal analysis.

Pediatr Pulmonol. 2017 Oct 30;:

Authors: McGarry ME, Neuhaus JM, Nielson DW, Burchard E, Ly NP

Abstract
BACKGROUND: Hispanic patients with cystic fibrosis (CF) have decreased life expectancy compared to non-Hispanic white patients. Pulmonary function is a main predictor of life expectancy in CF. Ethnic differences in pulmonary function in CF have been understudied. The objective was to compare longitudinal pulmonary function between Hispanic and non-Hispanic white patients with CF.
METHODS: This cohort study of 15 018 6-25 years old patients in the CF Foundation Patient Registry from 2008 to 2013 compared FEV1 percent predicted and longitudinal change in FEV1 percent predicted in Hispanic to non-Hispanic white patients. We used linear mixed effects models with patient-specific slopes and intercepts, adjusting for 14 demographic and clinical variables. We did sub-analyses by CFTR class, F508del copies, and PERT use.
RESULTS: Hispanic patients had lower FEV1 percent predicted (79.9%) compared with non-Hispanic white patients (85.6%); (-5.8%, 95%CI -6.7% to -4.8%, P < 0.001), however, there was no difference in FEV1 decline over time. Patients on PERT had a larger difference between Hispanic and non-Hispanic white patients in FEV1 percent predicted than patients not on PERT (-6.0% vs -4.1%, P = 0.02). The ethnic difference in FEV1 percent predicted was not statistically significant between CFTR classes (Class I-III: -6.1%, Class IV-V: -5.9%, Unclassified: -5.7%, P > 0.05) or between F508del copies (None: -7.6%, Heterozygotes: -5.6%, Homozygotes: -5.3%, P > 0.05).
CONCLUSIONS: Disparities in pulmonary function exist in Hispanic patients with CF early in life and then persist without improving or worsening over time. It is valuable to investigate the factors contributing to pulmonary function in Hispanic patients with CF.

PMID: 29082671 [PubMed - as supplied by publisher]

Anellovirus loads are associated with outcomes in pediatric lung transplantation.

Pediatr Transplant. 2017 Oct 29;:

Authors: Blatter JA, Sweet SC, Conrad C, Danziger-Isakov LA, Faro A, Goldfarb SB, Hayes D, Melicoff E, Schecter M, Storch G, Visner GA, Williams NM, Wang D

Abstract
Anelloviruses are DNA viruses ubiquitously present in human blood. Due to their elevated levels in immunosuppressed patients, anellovirus levels have been proposed as a marker of immune status. We hypothesized that low anellovirus levels, reflecting relative immunocompetence, would be associated with adverse outcomes in pediatric lung transplantation. We assayed blood samples from 57 patients in a multicenter study for alpha- and betatorquevirus, two anellovirus genera. The primary short-term outcome of interest was acute rejection, and longer-term outcomes were analyzed individually and as "composite" (death, chronic rejection, or retransplant within 2 years). Patients with low alphatorquevirus levels at 2 weeks post-transplantation were more likely to develop acute rejection within 3 months after transplant (P = .013). Low betatorquevirus levels at 6 weeks and 6 months after transplant were associated with death (P = .047) and the composite outcome (P = .017), respectively. There was an association between low anellovirus levels and adverse outcomes in pediatric lung transplantation. Alphatorquevirus levels were associated with short-term outcomes (ie, acute rejection), while betatorquevirus levels were associated with longer-term outcomes (ie, death, or composite outcome within 2 years). These observations suggest that anelloviruses may serve as useful biomarkers of immune status and predictors of adverse outcomes.

PMID: 29082660 [PubMed - as supplied by publisher]

Bone mineral density is related to lung function outcomes in young people with cystic fibrosis-A retrospective study.

Pediatr Pulmonol. 2017 Oct 30;:

Authors: Smith N, Lim A, Yap M, King L, James S, Jones A, Ranganathan S, Simm P

Abstract
INTRODUCTION: Improvements in the medical management of cystic fibrosis (CF) in recent years have resulted in increased prevalence of long-term sequelae of the condition, such as low bone mineral density (BMD) and hence an increased risk of fractures in later life. Aim To explore the interaction between BMD and lung function, nutrition, and genotype.
METHODS: This study was a retrospective audit of 202 children with CF from August 2000 to January 2016 to investigate associations between BMD Z-scores with clinical status, nutrition, and genetics using dual-energy absorptiometry X-ray data from the Royal Children's Hospital Melbourne, Australia.
RESULTS: Severity of both lung disease (P < 0.0001) and nutritional status (P < 0.05) was found to be strongly associated with BMD Z-scores.
CONCLUSIONS: This is the biggest study to date to provide further evidence that the severity of pulmonary disease is related to BMD in CF patients and therefore screening guidelines for bone health in children with CF should target individuals with the poorest clinical status.

PMID: 29082636 [PubMed - as supplied by publisher]