Overexpression of miRNA-25-3p inhibits Notch1 signaling and TGF-β-induced collagen expression in hepatic stellate cells.

Sci Rep. 2019 Jun 12;9(1):8541

Authors: Genz B, Coleman MA, Irvine KM, Kutasovic JR, Miranda M, Gratte FD, Tirnitz-Parker JEE, Olynyk JK, Calvopina DA, Weis A, Cloonan N, Robinson H, Hill MM, Al-Ejeh F, Ramm GA

Abstract
During chronic liver injury hepatic stellate cells (HSCs), the principal source of extracellular matrix in the fibrotic liver, transdifferentiate into pro-fibrotic myofibroblast-like cells - a process potentially regulated by microRNAs (miRNAs). Recently, we found serum miRNA-25-3p (miR-25) levels were upregulated in children with Cystic Fibrosis (CF) without liver disease, compared to children with CF-associated liver disease and healthy individuals. Here we examine the role of miR-25 in HSC biology. MiR-25 was detected in the human HSC cell line LX-2 and in primary murine HSCs, and increased with culture-induced activation. Transient overexpression of miR-25 inhibited TGF-β and its type 1 receptor (TGFBR1) mRNA expression, TGF-β-induced Smad2 phosphorylation and subsequent collagen1α1 induction in LX-2 cells. Pull-down experiments with biotinylated miR-25 revealed Notch signaling (co-)activators ADAM-17 and FKBP14 as miR-25 targets in HSCs. NanoString analysis confirmed miR-25 regulation of Notch- and Wnt-signaling pathways. Expression of Notch signaling pathway components and endogenous Notch1 signaling was downregulated in miR-25 overexpressing LX-2 cells, as were components of Wnt signaling such as Wnt5a. We propose that miR-25 acts as a negative feedback anti-fibrotic control during HSC activation by reducing the reactivity of HSCs to TGF-β-induced collagen expression and modulating the cross-talk between Notch, Wnt and TGF-β signaling.

PMID: 31189969 [PubMed - in process]

Cryo-electron microscopy of membrane proteins.

Methods. 2018 09 01;147:176-186

Authors: Thonghin N, Kargas V, Clews J, Ford RC

Abstract
Membrane proteins represent a large proportion of the proteome, but have characteristics that are problematic for many methods in modern molecular biology (that have often been developed with soluble proteins in mind). For structural studies, low levels of expression and the presence of detergent have been thorns in the flesh of the membrane protein experimentalist. Here we discuss the use of cryo-electron microscopy in breakthrough studies of the structures of membrane proteins. This method can cope with relatively small quantities of sample and with the presence of detergent. Until recently, cryo-electron microscopy could not deliver high-resolution structures of membrane proteins, but recent developments in transmission electron microscope technology and in the image processing of single particles imaged in the microscope have revolutionized the field, allowing high resolution structures to be obtained. Here we focus on the specific issues surrounding the application of cryo-electron microscopy to the study of membrane proteins, especially in the choice of a system to keep the protein soluble.

PMID: 29702228 [PubMed - indexed for MEDLINE]

Augmentation of CFTR function in human bronchial epithelial cells via SLC6A14-dependent amino acid uptake: Implications for treatment of Cystic Fibrosis.

Am J Respir Cell Mol Biol. 2019 Jun 12;:

Authors: Ahmadi S, Wu YS, Li M, Ip W, Lloyd-Kuzik A, Di Paola M, Du K, Xia S, Lew A, Bozoky Z, Forman-Kay J, Bear CE, Gonska T

Abstract
RATIONALE: SLC6A14 mediated L-arginine transport has been shown to augment the residual anion channel activity of the major mutant, F508del-CFTR, in the murine gastrointestinal tract. It is not yet known if this transporter augments residual and pharmacological corrected F508del-CFTR in primary airway epithelia.
OBJECTIVE: To determine the role of L-arginine uptake via SLC6A14 in modifying F508del-CFTR channel activity in airway cells from CF patients.
METHODS: Human bronchial epithelial cells from lung explants of non-CF (HBE) and CF patients (CF-HBE) were used for H3-flux, airway surface liquid (ASL) and Ussing chamber studies. We used α-methyltryptophan (α-MT) as specific inhibitor for SLC6A14. CFBE41o-, a commonly used CF airway cell line, was employed for studying the mechanism of the functional interaction between SLC6A14 and F508del-CFTR.
MAIN RESULTS: SLC6A14 is functionally expressed in CF-HBE cells. L-arginine uptake via SLC6A14 augmented F508del-CFTR function at baseline and after treatment with lumacaftor. SLC6A14 mediated L-arginine uptake also increased the ASL in CF-HBE cells. Using CFBE41o cells we showed that the positive SLC6A14 effect was mainly dependent on the nitric oxide synthase activity, nitrogen oxides including nitric oxide (NO) and phosphorylation by protein kinase G. These finding were confirmed in CF-HBE as iNOS inhibition abrogated the functional interaction between SLC6A14 and pharmacological corrected F508del-CFTR.
CONCLUSION: SLC6A14-mediated L-arginine transport augments residual F508del-CFTR channel function via a non-canonical, NO pathway. This effect is enhanced with increasing pharmacological rescue of F508del-CFTR to the membrane. The current study demonstrates how endogenous pathways can be utilized for the development of companion therapy in CF.

PMID: 31189070 [PubMed - as supplied by publisher]

Cystic fibrosis presentation in del. F508 and p. Tyr109Glyfs compound heterozygote CFTR state: a case report.

Croat Med J. 2019 Jun 13;60(3):246-249

Authors: Turkalj M, Matišić V, Šimić A, Juginović A, Erceg D, Tješić Drinković D, Höppner W, Primorac D

Abstract
The diagnosis of cystic fibrosis (CF) is commonly confirmed by molecular genetics with the presence of specific mutations of cystic fibrosis transmembrane conductance regulator (CFTR) gene. We report a case of cystic fibrosis (CF) in a 15-year-old female patient who is a compound heterozygote for CFTR gene, with delta F508 and Tyr109Glyfs mutations detected. This is the first detailed description of such a case in the medical literature. The primary CF presentation occurred at the age of 9 in the form of gastrointestinal symptoms including greasy, bulky, and foul-smelling stool. The patient exhibited delayed growth, with her height and weight being below the 5th centile for age according to the World Health Organization growth curves. Pancreatic enzyme supplement treatment was started immediately, alongside high-fat and high-calorie diet, resulting in patient's recovery and development. DNA analysis of CFTR gene demonstrated the presence of del. F508 mutation and a rare combining deletion and insertion mutation p. Tyr109Glyfs. The combination of the two mutations is very rare in CF patients and is therefore valuable to document this case in order to provide information on disease progression, therapy options, and outcomes. With standard treatment and early diagnosis, the patient is currently doing well and is not restricted by the disease in her daily and sports activities.

PMID: 31187952 [PubMed - in process]

Protein Misfolding Diseases and Therapeutic Approaches.

Curr Protein Pept Sci. 2019 Jun 09;:

Authors: Yadav K, Yadav A, Priyanka, Pandey VP, Dwivedi UN

Abstract
Protein folding is the process by which a polypeptide chain acquires its functional, native 3D structure. Protein misfolding, on the other hand, is a process in which proteins fails to fold into its native functional conformation. This misfolding of proteins may lead to precipitation of number of serious diseases such as Cystic fibrosis (CF), Alzheimer's disease (AD), Parkinson's disease (PD), and amyotrophic lateral sclerosis (ALS) etc. Protein quality-control (PQC) systems, consisting of molecular chaperones, proteases and regulatory factors, help in protein folding and prevent its aggregation. At the same time PQC syatems also do sorting and removal of improperly folded polypeptides. Among the major types of PQC systems involved in protein homeostasis are cytosolic, endoplasmic reticulum (ER) and mitochondrial ones. The cytosol PQC system includes a large number of component chaperones, such as nascent-polypeptide-associated complex (NAC), Hsp40, Hsp70, prefoldin and T Complex Protein-1 (TCP-1) ring complex (TRiC). Protein misfolding diseases caused due to defective cytosolic PQC system includes diseases involving keratin/collagen proteins, cardiomyopathies, phenylketonuria, PD and ALS. The components of PQC system of endoplasmic reticulum (ER) includes Binding immunoglobulin Protein (BiP), calnexin (CNX), calreticulin (CRT), glucose-regulated protein Grp94, the thiol-disulphide oxidoreductases, protein disulphide isomerase (PDI) and ERp57. ER-linked misfolding diseases include CF and familial neurohypophyseal diabetes insipidus (FNDI). The components of mitochondrial PQC system include mitochondrial chaperones such as the Hsp70, the Hsp60/Hsp10 and a set of proteases having AAA+ domains similar to the proteasome that are situated in the matrix or the inner membrane. Protein misfolding diseases caused due to defective mitochondrial PQC system include medium-chain acyl-CoA dehydrogenase (MCAD)/short-chain acyl-CoA dehydrogenase (SCAD) deficiency diseases, hereditary spastic paraplegia. Therapeutic approaches towards the treatment of various protein misfolding diseases using molecular, chemical and pharmacological classes of chaperones are also discussed.

PMID: 31187709 [PubMed - as supplied by publisher]

Exotoxin A-PLGA nanoconjugate vaccine against Pseudomonas aeruginosa infection: protectivity in murine model.

World J Microbiol Biotechnol. 2019 Jun 11;35(6):94

Authors: Safari Zanjani L, Shapouri R, Dezfulian M, Mahdavi M, Shafiee Ardestani M

Abstract
Pseudomonas aeruginosa is the major infectious agent of concern for cystic fibrosis (CF) patients. Therefore, it is necessary to develop appropriate strategies for preventing colonization by this bacterium and/or neutralizing virulence factors. In this study, we formulated the encapsulation of exotoxin A into PLGA nanoparticles. The biological activities of the nanovaccine candidate were also characterized. Based on the results, ETA-PLGA can act as a suitable immunogen to stimulate the humoral and cellular immune response. The antibodies raised against ETA-PLGA significantly decreased bacterial titer in the spleens of the immunized mice after challenge with PAO1 strain, compared to the control groups. The encapsulation of PLGA into ETA led to a significantly higher production of INF-γ, TNF-α, IL-4, and IL-17A cytokine responses compared to the ETA group. ETA-PLGA enhanced IgG responses in immunized mice compared to ETA antigen. We concluded that encapsulation of Pseudomonas aeruginosa ETA to PLGA nanoparticles can increase its functional activity by decreasing the bacterial dissemination.

PMID: 31187291 [PubMed - in process]

Novel AAV-mediated gene delivery system corrects CFTR function in pigs.

Am J Respir Cell Mol Biol. 2019 Jun 11;:

Authors: Cooney AL, Thornell IM, Singh BK, Shah VS, Stoltz DA, McCray PB, Zabner J, Sinn PL

Abstract
Cystic fibrosis (CF) is an autosomal recessive disease caused by a mutant cystic fibrosis transmembrane conductance regulator (CFTR) gene and characterized by chronic bacterial lung infections and inflammation. Complementation with functional CFTR normalizes anion transport across the airway surface. Adeno-associated virus (AAV) is a useful vector for gene therapy because of its low immunogenicity and ability to persist for months to years. However, episomal expression may decrease following cell division and ultimately require readministration. To overcome this, we designed an integrating AAV-based CFTR expressing vector termed PB/AAV carrying CFTR flanked by the terminal repeats (TRs) of the piggyBac transposon. With co-delivery of piggyBac transposase, PB/AAV can integrate into the host genome. Because of the AAV packaging constraints, careful consideration was required to ensure that the vector would package and express its CFTR cDNA cargo. In this short-term study, PB/AAV-CFTR was aerosolized to the airways of CF pigs in the absence of the transposase. Two weeks later, transepithelial Cl- current was restored in freshly excised tracheal and bronchus tissue. Additionally, we observed an increase in tracheal airway surface liquid (ASL) pH and bacterial killing as compared to untreated CF pigs. ASL from primary airway cells cultured from treated CF pigs exhibited increased pH correlating with decreased viscosity. Together, these results show that complementing CFTR in CF pigs with PB/AAV rescues the anion transport defect in a large animal CF model. Delivery of this integrating viral vector system to airway progenitor cells could lead to persistent, life-long expression in vivo.

PMID: 31184507 [PubMed - as supplied by publisher]

Hemoptysis in Pediatric Patients.

Cureus. 2019 Mar 23;11(3):e4305

Authors: Naum R, Speed B

Abstract
Hemoptysis is defined as the expectoration of blood or blood-tinged sputum. Blood-tinged sputum is a rare finding in the pediatric population. Finding the cause and treatment of the hemoptysis in pediatric patients is largely dependent on the history. In children, the most common causes of hemoptysis are infection and tracheostomy-related complications. Other causes include aberrant bronchial circulation, aspiration of foreign bodies, and bronchiectasis associated with cystic fibrosis. Due to the rarity of hemoptysis in pediatric patients, diagnosis and management of these patients can be difficult. It is important to refer to case reports and literature to best manage these patients. We report a case of a 3-year-old male patient who presented to the emergency department (ED) with a one-day history of hemoptysis. He presented with his adopted mother who was unable to provide a comprehensive past medical or family history other than stating that the patient has had recurrent bronchial infections since his adoption. She stated that the patient had only one episode of hemoptysis just prior to arrival. The patient did not appear to be in any respiratory distress and did not have any episodes of hemoptysis while in the ED. Due to his afebrile status and lack of evidence of current bleeding, the only intervention administered was an albuterol breathing treatment. He responded well to the breathing treatment and was discharged home with instructions to follow up with his primary care provider.

PMID: 31183285 [PubMed]

Pseudo-Bartter syndrome in children with cystic fibrosis.

Clin Case Rep. 2019 Jun;7(6):1123-1126

Authors: Faraji-Goodarzi M

Abstract
Cystic fibrosis (CF) is an autosomal recessive genetic disorder. We report a case of a boy aged 4.5 years with cystic fibrosis, presenting under-weightness, hypocalcemia, metabolic alkalosis, hypokalemia, and hyponatremia. Sweat analysis of the patients concluded pseudo-Bartter syndrome, which was successfully treated using antibiotics, physiotherapy, fluids, vitamin supplements, and pancreatic enzyme therapy.

PMID: 31183080 [PubMed]

Enhanced avidity from a multivalent fluorescent antimicrobial peptide enables pathogen detection in a human lung model.

Sci Rep. 2019 Jun 10;9(1):8422

Authors: Akram AR, Avlonitis N, Scholefield E, Vendrell M, McDonald N, Aslam T, Craven TH, Gray C, Collie DS, Fisher AJ, Corris PA, Walsh T, Haslett C, Bradley M, Dhaliwal K

Abstract
Rapid in situ detection of pathogens coupled with high resolution imaging in the distal human lung has the potential to provide new insights and diagnostic utility in patients in whom pneumonia is suspected. We have previously described an antimicrobial peptide (AMP) Ubiquicidin (fragment UBI29-41) labelled with an environmentally sensitive fluorophore that optically detected bacteria in vitro but not ex vivo. Here, we describe further chemical development of this compound and demonstrate that altering the secondary structure of the AMP to generate a tri-branched dendrimeric scaffold provides enhanced signal in vitro and ex vivo and consequently allows the rapid detection of pathogens in situ in an explanted human lung. This compound (NBD-UBIdend) demonstrates bacterial labelling specificity for a broad panel of pathogenic bacteria and Aspergillus fumigatus. NBD-UBIdend demonstrated high signal-to-noise fluorescence amplification upon target engagement, did not label host mammalian cells and was non-toxic and chemically robust within the inflamed biological environment. Intrapulmonary delivery of NBD-UBIdend, coupled with optical endomicroscopy demonstrated real-time, in situ detection of bacteria in explanted whole human Cystic Fibrosis lungs.

PMID: 31182770 [PubMed - in process]

Regulating, measuring and modelling the viscoelasticity of bacterial biofilms.

J Bacteriol. 2019 Jun 10;:

Authors: Charlton SGV, White MA, Jana S, Eland LE, Jayathilake PG, Burgess JG, Chen J, Wipat A, Curtis TP

Abstract
Biofilms occur in a broad range of environments with heterogeneous physico-chemical conditions, such as in bio-remediation plants, on surfaces of biomedical implants and in the lungs of cystic fibrosis patients. In these scenarios, biofilms are subjected to shear forces, but the mechanical integrity of these aggregates often prevents their disruption or dispersal. Their physical robustness is the result of the multiple bio-polymers secreted by constituent microbial cells which are also responsible for numerous biological functions. A better understanding of the role of these bio-polymers and their response to dynamic forces is therefore crucial for understanding the interplay between biofilm structure and function. In this manuscript, we review experimental techniques in rheology, which help quantify the viscoelasticity of biofilms, and modelling approaches from soft matter physics, that can assist our understanding of the rheological properties. We describe how these methods could be combined with synthetic biology approaches to control and investigate the effects of secreted polymers on the physical properties of biofilms. We argue that without an integrated approach of the three disciplines the links between genetics, composition and interaction of matrix biopolymers and the viscoelastic properties of biofilms will be much harder to uncover.

PMID: 31182499 [PubMed - as supplied by publisher]

Antisense oligonucleotide eluforsen is safe and improves respiratory symptoms in F508DEL cystic fibrosis.

J Cyst Fibros. 2019 Jun 07;:

Authors: Drevinek P, Pressler T, Cipolli M, De Boeck K, Schwarz C, Bouisset F, Boff M, Henig N, Paquette-Lamontagne N, Montgomery S, Perquin J, Tomkinson N, den Hollander W, Elborn JS

Abstract
BACKGROUND: Eluforsen is an antisense oligonucleotide designed to bind to the mRNA region around the F508-encoding deletion and restore the cystic fibrosis transmembrane conductance regulator (CFTR) protein function in the airway epithelium. We assessed the safety and tolerability, pharmacokinetics and exploratory measures of efficacy of inhaled eluforsen in cystic fibrosis (CF) patients homozygous for the F508del-CFTR mutation.
METHODS: This randomised, double-blind, placebo-controlled, dose escalation 1b study recruited adult CF subjects with a FEV1 > 70% predicted in four single ascending dose cohorts and four multiple ascending dose cohorts. Primary objectives were safety and tolerability. Secondary endpoints included pharmacokinetics, percent predicted forced expiratory volume in 1 s (ppFEV1), and Cystic Fibrosis Questionnaire-Revised (CFQ-R) Respiratory Symptom Score (RSS).
RESULTS: Single and multiple doses of inhaled eluforsen up to 50 mg were safe and well tolerated. A maximum tolerated dose was not established. Systemic exposure was low in all cohorts and lung function remained stable throughout the study. Three of four eluforsen-treated groups in the MAD study demonstrated an improvement in CFQ-R RSS at end of treatment with adjusted mean change from baseline values ranging from 6.4 to 12.7 points. In comparison, there was a mean decrease of 6.5 points in the placebo group from baseline to end of treatment.
CONCLUSIONS: Inhaled eluforsen up to 50 mg dosed 3 times per week for 4 weeks was safe and well tolerated, showed low systemic exposure, and demonstrated improvement in CFQ-R RSS, a relevant measure of clinical benefit in CF patients.

PMID: 31182369 [PubMed - as supplied by publisher]

Liver Ultrasound Patterns in Children with Cystic Fibrosis Correlate with Non-Invasive Tests of Liver Disease.

J Pediatr Gastroenterol Nutr. 2019 Jun 07;:

Authors: Ling SC, Ye W, Leung DH, Navarro OM, Weymann A, Karnsakul W, Freeman AJ, Magee JC, Narkewicz MR

Abstract
OBJECTIVES: Early identification of children with cystic fibrosis (CF) at risk for severe liver disease (CFLD) would enable targeted study of preventative therapies. There is no gold standard test for CFLD. Ultrasonography (US) is used to identify CFLD, but with concerns for its diagnostic accuracy. We aim to determine if differences in standard blood tests, imaging variables and non-invasive liver fibrosis indices correlate with liver US patterns and thus provide supportive evidence that a heterogeneous US liver pattern reflects clinically relevant liver disease.
METHODS: We studied baseline research abdominal US and bloodwork from 244 children with pancreatic insufficient CF, ages 3-12y, enrolled in a prospective study of the ability of US to predict CF cirrhosis (PUSH study). Children with a heterogeneous liver pattern on US (HTG, n = 62) were matched 1:2 in design with children with normal US (NL, n = 122). Analyses included children with nodular (NOD, n = 22) and homogeneous hyperechoic (HMG, n = 38) livers.
RESULTS: Univariate analysis showed significant differences between US groups for standard blood tests, spleen size, and non-invasive liver fibrosis indices. Multivariable models discriminated NOD vs. NL with excellent accuracy (AUROC 0.96). Models also distinguish HTG vs. NL (AUROC 0.76), NOD vs. HTG (0.78), and HMG vs NL (0.79).
CONCLUSION: Liver US patterns in children with CF correlate with platelet count, spleen size and indices of liver fibrosis. Multivariable models of these biomarkers have excellent discriminating ability for NL vs NOD, and good ability to distinguish other US patterns, suggesting that US patterns correlate with clinically relevant liver disease.

PMID: 31181021 [PubMed - as supplied by publisher]

New Algorithm for the Integration of Ultrasound into Cystic Fibrosis Liver Disease Screening.

J Pediatr Gastroenterol Nutr. 2019 Jun 07;:

Authors: Sellers ZM, Lee LW, Barth RA, Milla C

Abstract
OBJECTIVES: Liver nodularity occurs across the spectrum of cystic fibrosis liver disease (CFLD), from regenerative nodules to cirrhosis, and can occur without liver enzyme abnormalities. Our aims were to determine if incorporating abdominal ultrasound (US) with annual laboratory testing improves the detection of CFLD and establish CF-specific thresholds for liver screening labs.
METHODS: CF patients ≥6 years-old and exocrine pancreatic insufficient had an US with Doppler and shear wave elastography. Patients were divided into Normal, Echogenic, or Nodular groups, based on US findings. Results were compared to AST, ALT, Platelets, APRI (AST to Platelet Ratio Index), FIB-4 (Fibrosis-4), and GPR (GGT to Platelet Ratio). Receiver operator curve, sensitivity, specificity, positive predictive value, negative predictive value, and optimal cut-off with Youden's Index were calculated.
RESULTS: From 82 patients, incorporation of US identified more nodular livers than using labs alone. The Nodular group had significantly greater median AST (44), ALT (48), GGT (46), APRI (0.619), FIB-4 (0.286), GPR (1.431). Optimal cut-offs to detect liver nodularity in CF were AST >33, ALT >45, GGT >21, Platelets <230, APRI >0.367, FIB-4 >0.222, GPR >0.682. Using GGT, APRI, and GPR we generated an algorithm to direct the use of US in CFLD screening.
CONCLUSIONS: Using modified serum lab thresholds, addition of liver fibrosis indices, and/or abdominal US can increase detection of liver nodularity in CF. A combination of GGT, GPR, and APRI can help direct which CF children should undergo US evaluation. These tools may improve earlier identification of fibrosis and/or cirrhosis in CF patients.

PMID: 31181020 [PubMed - as supplied by publisher]

Lactation Consultation Sustains Breast Milk Intake in Infants with Cystic Fibrosis.

J Pediatr Gastroenterol Nutr. 2019 Jun 07;:

Authors: Miller T, Antos NJ, Brock LA, Wade T, Goday PS

Abstract
Guidelines recommend breastfeeding in cystic fibrosis (CF) but breastfeeding rates decline rapidly in CF. We initiated a quality improvement project to improve breastfeeding in CF by incorporating International Board-Certified Lactation Consultants (IBCLC) into the initial CF-diagnosis visit to support mothers who were breastfeeding at diagnosis. In the pre-intervention group, only 8/14 (57%) continued to provide BM after the 1st visit, while post-intervention, 16/17 (94%) mothers continued to do so. (p = 0.03). The duration of any (or partial) breastfeeding increased to an average of 7.7 months from an average of 6.4 months pre-intervention (p = 0.45). The weight z score and weight-for-length z score at diagnosis showed no change at 6 or 12 months (all comparisons p value > 0.05). We conclude that mothers who met with the IBCLC were less likely to quit breastfeeding and hypothesize that lactation support to mothers can prolong the duration of breastfeeding in infants with CF.

PMID: 31181018 [PubMed - as supplied by publisher]

Randomised cross-over trial evaluating the short-term effects of non-invasive ventilation as an adjunct to airway clearance techniques in adults with cystic fibrosis.

BMJ Open Respir Res. 2019;6(1):e000399

Authors: Stanford G, Parrott H, Bilton D, Agent P, Banya W, Simmonds N

Abstract
Introduction: Non-invasive ventilation (NIV) is used in cystic fibrosis (CF) to support airway clearance techniques (ACTs) by augmenting tidal volumes and reducing patient effort. However, the evidence base for this is limited. We hypothesised that NIV, in addition to usual ACT, would increase sputum clearance. In addition, we investigated ease of sputum clearance (EoC), work of breathing (WoB) and NIV tolerability.
Methods: Adults with CF (16+ years) at the end of hospitalisation for a pulmonary exacerbation were randomised to a cross-over trial of NIV-supported ACT or ACT alone in two consecutive days. No other changes to standard care were made. The primary outcome was the total 24-hour expectorated sputum wet weight after the intervention. Spirometry was completed pre-treatment and post-treatment. Oxygen saturations were measured pre-treatment, during treatment and post-treatment. EoC and WoB were assessed using Visual Analogue Scale.
Results: 14 subjects completed the study (7 male, mean age 35 [SD 17] years, mean forced expiratory volume in 1 s [FEV1] 49 [20] % predicted). The difference between treatment regimens was -0.98 g sputum (95% CI -11.5 to 9.6, p=0.84) over 24 hours. During treatment oxygen saturations were significantly higher with NIV-supported ACT (mean difference 2.0, 95% CI 0.9 to 2.6, p=0.0004). No other significant differences were found in post-treatment FEV1, EoC, WoB, oxygen saturations or subject preference.
Conclusions: There was no difference in treatment effect between NIV-supported ACT and ACT alone, although the study was underpowered. Oxygen saturations were significantly higher during NIV-supported ACT, but with no effect on post-treatment saturations. NIV was well tolerated.
Trial registration number: NCT01885650.

PMID: 31179002 [PubMed]

Inhaled hypertonic saline for 3-6-year-olds with cystic fibrosis.

Lancet Respir Med. 2019 Jun 06;:

Authors: Southern KW, Sinha IP

PMID: 31178424 [PubMed - as supplied by publisher]

Inhaled hypertonic saline in preschool children with cystic fibrosis (SHIP): a multicentre, randomised, double-blind, placebo-controlled trial.

Lancet Respir Med. 2019 Jun 06;:

Authors: Ratjen F, Davis SD, Stanojevic S, Kronmal RA, Hinckley Stukovsky KD, Jorgensen N, Rosenfeld M, SHIP Study Group

Abstract
BACKGROUND: Inhaled hypertonic saline enhances mucociliary clearance, improves lung function, and reduces pulmonary exacerbations in people with cystic fibrosis older than age 6 years. We aimed to assess the effect of inhaled hypertonic saline on the lung clearance index (LCI2·5)-a measure of ventilation inhomogeneity-in children aged 3-6 years with cystic fibrosis.
METHODS: The Saline Hypertonic in Preschoolers (SHIP) Study was a randomised, double-blind, placebo-controlled trial at 25 cystic fibrosis centres in Canada and the USA. Eligible participants were aged 36-72 months; had a confirmed diagnosis of cystic fibrosis; were able to comply with medication use, study visits, and study procedures; and were able to complete at least two technically acceptable trials of multiple breath washout (MBW). Participants were randomly assigned (1:1) via a web-based data entry system that confirmed enrolment eligibility to inhaled 7% hypertonic saline or 0·9% isotonic saline nebulised twice daily (for no more than 15 min per dose) for 48 weeks. Permuted block randomisation was done separately for participants aged 36-54 months and those aged 55-72 months to ensure approximate balance by treatment group in the two age groups. The primary endpoint was the change in the LCI2·5 measured by nitrogen MBW from baseline to week 48. All study sites were trained and certified in MBW. Analysis was by intention to treat. This study is registered with Clinicaltrials.gov, number NCT02378467.
FINDINGS: Between April 21, 2015, and Aug 4, 2017, 150 participants were enrolled and randomly assigned, 76 to the hypertonic saline group and 74 to the isotonic saline group. Overall 89% of the MBW tests produced acceptable data. At 48 weeks, treatment with hypertonic saline was associated with a significant decrease (ie, improvement) in LCI2·5 compared with isotonic saline (mean treatment effect -0·63 LCI2·5 units [95% CI -1·10 to -0·15]; p=0·010). Six participants in the hypertonic saline group had ten serious adverse events and eight participants in the isotonic saline group had nine serious adverse events. The serious adverse events reported were cough (two patients [3%] in the hypertonic saline group vs three [4%] in the isotonic saline group), gastrostomy tube placement or rupture (two [3%] vs one [1%]), upper gastrointestinal disorders (one [1%] vs two [3%]), distal intestinal obstruction syndrome (one [1%] vs one [1%]), and decreased pulmonary function (none vs one [1%]). None of these serious adverse events was judged to be treatment related.
INTERPRETATION: Inhaled hypertonic saline improved the LCI2·5 in children aged 3-6 years, and could be a suitable early intervention in cystic fibrosis.
FUNDING: Cystic Fibrosis Foundation.

PMID: 31178421 [PubMed - as supplied by publisher]

Type I interferon induced by TLR2-TLR4-MyD88-TRIF-IRF3 controls Mycobacterium abscessus subsp. abscessus persistence in murine macrophages via nitric oxide.

Int J Med Microbiol. 2019 Jun 01;:

Authors: Ruangkiattikul N, Rys D, Abdissa K, Rohde M, Semmler T, Tegtmeyer PK, Kalinke U, Schwarz C, Lewin A, Goethe R

Abstract
Mycobacterium abscessus (MAB) is an emerging, rapidly growing non-tuberculous Mycobacterium causing therapy-resistant pulmonary disease especially in patients with cystic fibrosis (CF). Smooth and rough colony type MAB can be isolated from infected patients whereby rough colony type MAB are more often associated with severe disease. Disease severity is also associated with an alternated type I interferon (IFN-I) response of the MAB-infected patients. However the relevance of this response for the outcome of MAB infection is still unknown. In this study, we analyzed the IFNβ expression of murine macrophages infected with a MAB rough colony strain (MAB-R) isolated from a patient with progressive CF and compared it to macrophages infected with the MAB smooth colony type reference strain (MAB-S). We found that MAB-R infected macrophages expressed significantly more IFNβ mRNA and protein than MAB-S infected macrophages. Higher IFNβ induction by MAB-R was associated with higher TNF expression and intracellular killing while low IFNβ induction was associated with lower TNF expression and persistence of MAB-S. IFNβ induction was independent of the intracellular cGAS-STING recognition pathway. MAB appeared to be recognized extracellularly and induced IFNβ expression via TLR2-TLR4-MyD88-TRIF-IRF3 dependent pathways. By using macrophages lacking the IFN-I receptor we demonstrate that MAB induced IFN-I response essentially contributed to restricting MAB-R and MAB-S infections by activating macrophage Nos2 expression and nitric oxide production. Thus IFN-I seem to influence the intrinsic ability of macrophages to control MAB infections. As MAB persists over long time periods in susceptible patients, our findings suggest that virulence of MAB strains is promoted by an insufficient IFN-I response of the host.

PMID: 31178418 [PubMed - as supplied by publisher]

Study of a Cohort of Patients With Cystic Fibrosis and Isolation of Scedosporium spp.

Arch Bronconeumol. 2019 Jun 06;:

Authors: Erro Iribarren M, Girón Moreno RM, Diab Cáceres L, Pastor Sanz MT, Buendía Moreno B, Alarcón Cavero T, Granja Torrecillas S, Zurita Cruz ND, Ancochea Bermúdez J

Abstract
INTRODUCTION: In recent years an increase in the prevalence of colonization and infection by Scedosporium spp. in patients with cystic fibrosis (CF) has been observed. In this article, we study the frequency of isolation of Scedosporium spp. in an adult CF Unit, analyzing characteristics of the patients and predisposing factors.
METHODS: A retrospective observational study was conducted in 87 adult CF patients in whom the presence of positive culture for Scedosporium spp. was tested for a 5-year period (January 2012-July 2017). We recorded the following clinical variables: age, sex, body mass index, genotype, presence of pancreatic insufficiency, bacterial colonization, lung function, other complications, exacerbations and treatment, and the modified Bhalla score from the last high-resolution computed tomography. Results were analyzed with IBM SPSS Statistics Version 22.0 software.
RESULTS: Scedosporium spp. was isolated in 25.3% of patients. In the bivariate analysis, these patients showed a higher rate of Pseudomonas aeruginosa infection, worse score in the Bhalla classification (highlighting the following items: bronchiectasis, mucus plugs and bronchial generations), a slight decrease in the lung diffusion capacity and more frequently received inhaled antibiotics. In the logistic regression multivariate analysis, only the bronchial generations item was significant.
CONCLUSION: Scedosporium spp. must be considered an emerging opportunistic pathogen in patients with CF whose clinical involvement, risk factors or need for treatment is unknown.

PMID: 31178266 [PubMed - as supplied by publisher]