The role of non-invasive modalities for assessing inflammation in patients with non-cystic fibrosis bronchiectasis.

Cytokine. 2017 Aug 29;:

Authors: Tsikrika S, Dimakou K, Papaioannou AI, Hillas G, Thanos L, Kostikas K, Loukides S, Papiris S, Koulouris N, Bakakos P

Abstract
INTRODUCTION: Bronchiectasis is a heterogeneous entity, taking into account clinical characteristics, inflammatory response, effectiveness of treatment and frequency of exacerbations. In stable state non-cystic fibrosis (non-CF) bronchiectasis, little is known about non-invasive techniques used for evaluating airway inflammation in obstructive airway diseases.
OBJECTIVES: We sought to evaluate the associations between induced sputum and clinical/radiologic characteristics, and the differences between biomarkers expressing Th1 and Th2 response in patients with non-CF bronchiectasis and to compare our findings with a previously studied population of patients with asthma and COPD.
METHODS: We evaluated prospectively collected data from subjects with bronchiectasis. Comparisons were made between clinical, radiographic and physiologic characteristics, as well as induced sputum markers using appropriate statistical tools. We compared the levels of sputum markers with those of a previously studied cohort of asthma and COPD patients.
RESULTS: We enrolled 40 subjects (21men, mean age 63.5yrs) with bronchiectasis. Fifteen subjects (37.5%) had a neutrophilic phenotype, 7 (17.5%) had an eosinophilic phenotype, 3 (12.5%) had a mixed neutrophilic-eosinophilic phenotype and 15 (37.5%) had a paucigranulocytic phenotype. Subjects with sputum neutrophilia had more severe bronchiectasis in HRCT and higher levels of IL-8 in sputum, whereas subjects with eosinophilia had higher levels of FeNO, greater bronchodilator reversibility and higher sputum IL-13. Sputum IL-8 levels were higher in subjects exhibiting frequent exacerbations and correlated with neutrophils in sputum (r=0.799), the extent of bronchiectasis in HRCT (r=0.765) and post-bronchodilator FEV1 (r=-0.416). Sputum IL-13 levels correlated with sputum eosinophils (r=0.656) and bronchodilator reversibility (r=0.441). Neutrophilic bronchiectasis exhibited comparable IL-8 levels to COPD, whereas eosinophilic bronchiectasis showed significantly lower IL-13 levels compared to asthma.
CONCLUSIONS: Sputum cell counts and IL-8 and IL-13 correlate with distinct clinical and functional measurements of disease severity and therefore may have a role for non-invasively assessing inflammation in non-cystic fibrosis bronchiectasis.

PMID: 28863927 [PubMed - as supplied by publisher]

Cas9/gRNA targeted excision of cystic fibrosis-causing deep-intronic splicing mutations restores normal splicing of CFTR mRNA.

PLoS One. 2017;12(9):e0184009

Authors: Sanz DJ, Hollywood JA, Scallan MF, Harrison PT

Abstract
Cystic Fibrosis is an autosomal recessive disorder caused by mutations in the CFTR gene. CRISPR mediated, template-dependent homology-directed gene editing has been used to correct the most common mutation, c.1521_1523delCTT / p.Phe508del (F508del) which affects ~70% of individuals, but the efficiency was relatively low. Here, we describe a high efficiency strategy for editing of three different rare CFTR mutations which together account for about 3% of individuals with Cystic Fibrosis. The mutations cause aberrant splicing of CFTR mRNA due to the creation of cryptic splice signals that result in the formation of pseudoexons containing premature stop codons c.1679+1634A>G (1811+1.6kbA>G) and c.3718-2477C>T (3849+10kbC>T), or an out-of-frame 5' extension to an existing exon c.3140-26A>G (3272-26A>G). We designed pairs of Cas9 guide RNAs to create targeted double-stranded breaks in CFTR either side of each mutation which resulted in high efficiency excision of the target genomic regions via non-homologous end-joining repair. When evaluated in a mini-gene splicing assay, we showed that targeted excision restored normal splicing for all three mutations. This approach could be used to correct aberrant splicing signals or remove disruptive transcription regulatory motifs caused by deep-intronic mutations in a range of other genetic disorders.

PMID: 28863137 [PubMed - in process]

Another Brick in the Wall: Lung Clearance Index and Lower Airways Pathology in Preschool Cystic Fibrosis.

Ann Am Thorac Soc. 2017 Sep;14(9):1389-1390

Authors: Rayment JH, Ratjen F

PMID: 28862497 [PubMed - in process]

Association between lung function, physical activity level and postural evaluation variables in adult patients with cystic fibrosis.

Clin Respir J. 2017 Sep 01;:

Authors: Cherobin IA, Dalcin PTR, Ziegler B

Abstract
INTRODUCTION: With the evolution of the cystic fibrosis (CF) disease, the decline of lung function associated with metabolic disorders and malnutrition, causes alterations in respiratory mechanics, musculoskeletal disorders and thoracic deformities, bringing injury to the individual's quality of life.
OBJECTIVE: To verify the association between lung function, physical activity level and postural evaluation variables in adults with CF.
METHODS: All patients underwent clinical evaluation and spirometry. The International Physical Activity Questionnaire (IPAQ) and an accelerometer were used to verify the physical activity level (PA). Photogrammetry was used with the aid of the Software of postural evaluation (SAPO) and, for complementary comparisons, the 6-minute walk test (6MWT) was used.
RESULTS: Twenty-eight adult subjects with CF, mean age of 25.1 ± 6.3 years and mean expiratory volume in the first second (FEV1 ) of 47.1 ± 20.9% of the predicted group participated in the study. The FEV1 correlated with the parameters obtained by the accelerometer (r = 0.723, p = 0.000), postural evaluation (r = -0.483, p = 0.005) and 6MWT (r = 0.439, p = 0.019), but there was no correlation with the data obtained by IPAQ (r = -0.282; p = 0.073). The time in which each individual remained in moderate to vigorous physical activity correlated with parameters of postural evaluation (thoracic kyphosis r = -0.484, p = 0.031, cervical lordosis r = 0.531, p = 0.016), 6MWT (r = 0.564; p = 0.010) and with the total METS obtained by IPAQ (r = 0.451, p = 0.046).
CONCLUSION: Lung function in patients with CF disease is associated with higher thoracic kyphosis, shorter time in moderate and vigorous PA, and shorter distance covered in 6MWT. The accelerometer has been shown to be the best instrument for assessing PA in this public. This article is protected by copyright. All rights reserved.

PMID: 28862396 [PubMed - as supplied by publisher]

Early follow-up of lung disease in infants with cystic fibrosis using the raised volume rapid thoracic compression technique and computed tomography during quiet breathing.

Pediatr Pulmonol. 2017 Sep 01;:

Authors: Gauthier R, Cabon Y, Giroux-Metges MA, Du Boisbaudry C, Reix P, Bourgeois ML, Chiron R, Molinari N, Saguintaah M, Amsallem F, Matecki S

Abstract
BACKGROUND: Among the different techniques used to monitor lung disease progression in infants with CF diagnosed by Newborn screening (NBS), raised volume-rapid thoracic compression (RVRTC) remains a promising tool. However, the need of sedation and positive pressure ventilation considerably limits its clinical use. We recently described a semi-quantitative method to evaluate air trapping by chest tomography during quite breathing without sedation (CTqb score). This parameter is the radiological sign of airway obstruction and could be also used for lung disease follow-up in infants with CF. However, its discriminative power compared with RVRTC and correlation with lung function parameters are not known.
OBJECTIVES: To compare the discriminative powers of the CTqb score and RVRTC parameters and to determine their correlation during the first year of life of infants with CF.
METHODS: In this multicenter longitudinal study, infants with CF diagnosed by NBS underwent RVRTC and CT during quite breathing at 10 ± 4 weeks (n = 30) and then at 13 ± 1 months of age (n = 28).
RESULTS: All RVRTC parameters and the CTqb score remained stable between evaluations. The CTqb score showed a higher discriminative power than forced expiratory volume in 0.5 s (FEV0.5 ; the main RVRTC parameter) at both visits (66% and 50% of abnormal values vs 30% and 28%, respectively). No correlation was found between CTqb score and, the different RVRTC parameters or the plethysmographic functional residual capacity, indicating that they evaluate different aspect of CF lung disease.

PMID: 28861941 [PubMed - as supplied by publisher]

Assessment of selected media supplements to improve F/HN lentiviral vector production yields.

Sci Rep. 2017 Aug 31;7(1):10198

Authors: Gélinas JF, Davies LA, Gill DR, Hyde SC

Abstract
The development of lentiviral-based therapeutics is challenged by the high cost of current Good Manufacturing Practices (cGMP) production. Lentiviruses are enveloped viruses that capture a portion of the host cell membrane during budding, which then constitutes part of the virus particle. This process might lead to lipid and protein depletion in the cell membrane and affect cell viability. Furthermore, growth in suspension also causes stresses that can affect virus production yields. To assess the impact of these issues, selected supplements (Cholesterol Lipid Concentrate, Chemically Defined Lipid Concentrate, Lipid Mixture 1, Gelatin Peptone N3, N-Acetyl-L-Cysteine and Pluronic F-68) were assayed in order to improve production yields in a transient transfection production of a Sendai virus F/HN-pseudotyped HIV-1-based third generation lentiviral vector in FreeStyle 293 (serum-free media) in suspension. None of the supplements tested had a significant positive impact on lentiviral vector yields, but small non-significant improvements could be combined to increase vector production in a cell line where other conditions have been optimised.

PMID: 28860488 [PubMed - in process]

The endothelium in hypoxic pulmonary vasoconstriction.

J Appl Physiol (1985). 2017 Aug 31;:jap.00120.2017

Authors: Grimmer B, Kuebler WM

Abstract
Hypoxic pulmonary vasoconstriction (HPV) in combination with hypercapnic pulmonary vasoconstriction redistributes pulmonary blood flow from poorly aerated to better ventilated lung regions by an active process of local vasoconstriction. Impairment of HPV results in ventilation-perfusion mismatch and is commonly associated with various lung diseases including pneumonia, sepsis, or cystic fibrosis. Although several regulatory pathways have been identified, considerable knowledge gaps persist, and a unifying concept of the signaling pathways that underlie HPV and their impairment in lung diseases has not yet emerged. In the past, conceptual models of HPV have focused on pulmonary arterial smooth muscle cells (PASMC) acting as sensor and effector of hypoxia in the pulmonary vasculature. In contrast, the endothelium was considered a modulating bystander in this scenario. For an ideal design, however, the oxygen sensor in HPV should be located in the region of gas exchange, i.e. in the alveolar capillary network. This concept requires the retrograde propagation of the hypoxic signal along the endothelial layer of the vascular wall and subsequent contraction of PASMC in upstream arterioles that is elicited via a temporospatially tightly controlled endothelial-smooth muscle cell crosstalk. The present review summarizes recent work that provides proof-of-principle for the existence and functional relevance of such signaling pathway in HPV that involves important roles for connexin 40, epoxyeicosatrienoic acids, sphingolipids, and cystic fibrosis transmembrane conductance regulator. Of translational relevance, implication of these molecules provides for novel mechanistic explanations for impaired ventilation/perfusion matching in patients with pneumonia, sepsis, cystic fibrosis and presumably various other lung diseases.

PMID: 28860164 [PubMed - as supplied by publisher]

MUC5B mucin bundles from the submucosal glands coated with the MUC5AC mucin clear normal trachea.

Biochem Biophys Res Commun. 2017 Aug 28;:

Authors: Ermund A, Meiss LN, Rodriguez-Pineiro AM, Bähr A, Nilsson HE, Trillo-Muyo S, Ridley C, Thornton DJ, Wine JJ, Hebert H, Klymiuk N, Hansson GC

Abstract
To understand the mucociliary clearance system, mucins were visualized by light, confocal and electron microscopy, and mucus was stained by Alcian blue and tracked by video microscopy on tracheal explants of newborn piglets. We observed long linear mucus bundles that appeared at the submucosal gland openings and were transported cephalically. The mucus bundles were shown by mass spectrometry and immunostaining to have a core made of MUC5B mucin and were coated with MUC5AC mucin produced by surface goblet cells. The transport speed of the bundles was slower than the airway surface liquid flow. We suggest that the goblet cell MUC5AC mucin anchors the mucus bundles and thus controls their transport. Normal clearance of the respiratory tree of pigs and humans, both rich in submucosal glands, is performed by thick and long mucus bundles.

PMID: 28859985 [PubMed - as supplied by publisher]

Sinus bacteriology in patients with cystic fibrosis or primary ciliary dyskinesia: A systematic review.

Am J Rhinol Allergy. 2017 Sep 01;31(5):293-298

Authors: Møller ME, Alanin MC, Grønhøj C, Aanæs K, Høiby N, von Buchwald C

Abstract
BACKGROUND: A correlation exists between the microbial flora of the upper and lower airways in patients with cystic fibrosis (CF) or with primary ciliary dyskinesia (PCD). The sinuses can function as a bacterial reservoir where gram-negative bacteria adapt to the airways and repeatedly are aspirated to and colonize the lungs according to the theory of the united (unified) airways. Whereas the pattern of bacterial flora in the lower airways has been extensively studied, the upper airways have drawn limited attention.
OBJECTIVE: Our aim was to review the literature that reported bacterial flora in the sinuses and nasal cavities of patients with CF or PCD.
METHODS: A number of medical literature data bases were systematically searched between January 1960 and July 2016. We applied the following inclusion criteria: a minimum of one case of PCD (or Kartagener syndrome) or CF, and microbiology analyses from the nose or paranasal sinuses.
RESULTS: We included 46 studies (1823 patients) from 16 countries. Staphylococcus aureus was found in 30% of the noses and sinuses of patients with CF. Other common bacteria found included Pseudomonas aeruginosa, coagulase negative staphylococci, and Haemophilus influenzae. In PCD, H. influenzae was the most common bacteria (28%), followed by Streptococcus pneumoniae and P. aeruginosa. If studies that included nonsurgical swab and blowing samples were excluded, then P. aeruginosa was the most common bacterium in patients with CF (34%) and in patients with PCD (50%), followed by S. aureus and H. influenza.
CONCLUSION: S. aureus, P. aeruginosa, coagulase negative staphylococci, and H. influenzae dominated in the upper airways of patients with CF. In patients with PCD, H. influenzae, S. pneumoniae, and P. aeruginosa dominated. When studies that included swab and blowing samples were excluded, P. aeruginosa was the most common bacterium in both groups. Direct comparisons among the studies were restricted due to very heterogeneous methods, and a better standardization of procedures and outcomes is needed.

PMID: 28859703 [PubMed - in process]

The role of multispecies social interactions in shaping Pseudomonas aeruginosa pathogenicity in the cystic fibrosis lung.

FEMS Microbiol Lett. 2017 Aug 15;364(15):

Authors: O'Brien S, Fothergill JL

Abstract
Pseudomonas aeruginosa is a major pathogen in the lungs of cystic fibrosis (CF) patients. However, it is now recognised that a diverse microbial community exists in the airways comprising aerobic and anaerobic bacteria as well as fungi and viruses. This rich soup of microorganisms provides ample opportunity for interspecies interactions, particularly when considering secreted compounds. Here, we discuss how P. aeruginosa-secreted products can have community-wide effects, with the potential to ultimately shape microbial community dynamics within the lung. We focus on three well-studied traits associated with worsening clinical outcome in CF: phenazines, siderophores and biofilm formation, and discuss how secretions can shape interactions between P. aeruginosa and other commonly encountered members of the lung microbiome: Staphylococcus aureus, the Burkholderia cepacia complex, Candida albicans and Aspergillus fumigatus. These interactions may shape the evolutionary trajectory of P. aeruginosa while providing new opportunities for therapeutic exploitation of the CF lung microbiome.

PMID: 28859314 [PubMed - in process]

Detection of misidentifications of species from the Burkholderia cepacia complex and description of a new member, the soil bacterium Burkholderia catarinensis sp. nov.

Pathog Dis. 2017 Aug 31;75(6):

Authors: Bach E, Sant'Anna FH, Magrich Dos Passos JF, Balsanelli E, de Baura VA, Pedrosa FO, de Souza EM, Passaglia LMP

Abstract
The correct identification of bacteria from the Burkholderia cepacia complex (Bcc) is crucial for epidemiological studies and treatment of cystic fibrosis infections. However, genome-based identification tools are revealing many controversial Bcc species assignments. The aim of this work is to re-examine the taxonomic position of the soil bacterium B. cepacia 89 through polyphasic and genomic approaches. recA and 16S rRNA gene sequence analysis positioned strain 89 inside the Bcc group. However, based on the divergence score of seven concatenated allele sequences, and values of average nucleotide identity, and digital DNA:DNA hybridization, our results suggest that strain 89 is different from other Bcc species formerly described. Thus, we propose to classify Burkholderia sp. 89 as the novel species Burkholderia catarinensis sp. nov. with strain 89T (=DSM 103188T = BR 10601T) as the type strain. Moreover, our results call the attention to some probable misidentifications of Bcc genomes at the National Center for Biotechnology Information database.

PMID: 28859310 [PubMed - in process]

Release from Xenopus oocyte prophase I meiotic arrest is independent of a decrease in cAMP levels or PKA activity.

Development. 2016 Jun 01;143(11):1926-36

Authors: Nader N, Courjaret R, Dib M, Kulkarni RP, Machaca K

Abstract
Vertebrate oocytes arrest at prophase of meiosis I as a result of high levels of cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) activity. In Xenopus, progesterone is believed to release meiotic arrest by inhibiting adenylate cyclase, lowering cAMP levels and repressing PKA. However, the exact timing and extent of the cAMP decrease is unclear, with conflicting reports in the literature. Using various in vivo reporters for cAMP and PKA at the single-cell level in real time, we fail to detect any significant changes in cAMP or PKA in response to progesterone. More interestingly, there was no correlation between the levels of PKA inhibition and the release of meiotic arrest. Furthermore, we devised conditions whereby meiotic arrest could be released in the presence of sustained high levels of cAMP. Consistently, lowering endogenous cAMP levels by >65% for prolonged time periods failed to induce spontaneous maturation. These results argue that the release of oocyte meiotic arrest in Xenopus is independent of a reduction in either cAMP levels or PKA activity, but rather proceeds through a parallel cAMP/PKA-independent pathway.

PMID: 27122173 [PubMed - indexed for MEDLINE]

Lung transplantation in cystic fibrosis patients with difficult to treat lung infections.

Curr Opin Pulm Med. 2017 Aug 30;:

Authors: Dupont L

Abstract
PURPOSE OF REVIEW: In cystic fibrosis (CF) patients with end-stage pulmonary disease, lung transplantation (LTx) remains a life-extending therapy with good outcome in most patients. Despite early concern about chronic pretransplantation infections in the context of posttransplantation immunosuppression, typical CF-associated organisms such as Pseudomonas aeruginosa turned out to be quite well manageable and associated with favorable outcomes in transplanted CF patients, even in patients with highly resistant strains. However, the situation is less evident with other pathogens.
RECENT FINDINGS: Burkholderia cenocepacia is associated with reduced survival and regarded as a contraindication for LTx in most centers, other Burkholderia species are less problematic. Other resistant Gram-negative bacteria and methicillin-resistant staphylococcus aureus in CF patients are not regarded as a contraindication. Nontuberculous mycobacteria disease in CF patients does not preclude successful recovery after LTx, although postoperative complications can be expected in patients with Mycobacterium abscessus and specific management is indicated. Fungal species should be treated aggressively to limit morbidity after transplantation.
SUMMARY: Despite its complexity, LTx is safe in most CF patients, with good outcomes if the pathogens that are present are identified and adequately treated.

PMID: 28858970 [PubMed - as supplied by publisher]

Inflammatory Diseases and Growth: Effects on the GH-IGF Axis and on Growth Plate.

Int J Mol Sci. 2017 Aug 31;18(9):

Authors: Cirillo F, Lazzeroni P, Sartori C, Street ME

Abstract
This review briefly describes the most common chronic inflammatory diseases in childhood, such as cystic fibrosis (CF), inflammatory bowel diseases (IBDs), juvenile idiopathic arthritis (JIA), and intrauterine growth restriction (IUGR) that can be considered, as such, for the changes reported in the placenta and cord blood of these subjects. Changes in growth hormone (GH) secretion, GH resistance, and changes in the insulin-like growth factor (IGF) system are described mainly in relationship with the increase in nuclear factor-κB (NF-κB) and pro-inflammatory cytokines. Changes in the growth plate are also reported as well as a potential role for microRNAs (miRNAs) and thus epigenetic changes in chronic inflammation. Many mechanisms leading to growth failure are currently known; however, it is clear that further research in the field is still warranted.

PMID: 28858208 [PubMed - in process]

A double echo ultra short echo time (UTE) acquisition for respiratory motion-suppressed high resolution imaging of the lung.

Magn Reson Med. 2017 Aug 30;:

Authors: Delacoste J, Chaptinel J, Beigelman-Aubry C, Piccini D, Sauty A, Stuber M

Abstract
PURPOSE: Magnetic resonance imaging is a promising alternative to computed tomography for lung imaging. However, organ motion and poor signal-to-noise ratio, arising from short T2*, impair image quality. To alleviate these issues, a new retrospective gating method was implemented and tested with an ultra-short echo time sequence.
METHODS: A 3D double-echo ultra-short echo time sequence was used to acquire data during free breathing in ten healthy adult subjects. A self-gating method was used to reconstruct respiratory motion suppressed expiratory and inspiratory images. These images were objectively compared to uncorrected data sets using quantitative end-points (pulmonary vessel sharpness, lung-liver interface definition, signal-to-noise ratio). The method was preliminarily tested in two cystic fibrosis patients who underwent computed tomography.
RESULTS: Vessel sharpness in expiratory ultra-short echo time data sets with second echo motion detection was significantly higher (13% relative increase) than in uncorrected images while the opposite was observed in inspiratory images. The method was successfully applied in patients and some findings (e.g., hypointense areas) were similar to those from computed tomography.
CONCLUSION: Free breathing ultra-short echo time was successfully implemented, allowing flexible image reconstruction of two different respiratory states. Objective improvements in image quality were obtained with the new method and initial feasibility in a clinical setting was demonstrated. Magn Reson Med, 2017. © 2017 International Society for Magnetic Resonance in Medicine.

PMID: 28856720 [PubMed - as supplied by publisher]

Feasibility of quantitative regional ventilation and perfusion mapping with phase-resolved functional lung (PREFUL) MRI in healthy volunteers and COPD, CTEPH, and CF patients.

Magn Reson Med. 2017 Aug 30;:

Authors: Voskrebenzev A, Gutberlet M, Klimeš F, Kaireit TF, Schönfeld C, Rotärmel A, Wacker F, Vogel-Claussen J

Abstract
PURPOSE: In this feasibility study, a phase-resolved functional lung imaging postprocessing method for extraction of dynamic perfusion and ventilation parameters using a conventional 1H lung MRI Fourier decomposition acquisition is introduced.
METHODS: Time series of coronal gradient-echo MR images with a temporal resolution of 288 to 324 ms of two healthy volunteers, one patient with chronic thromboembolic hypertension, one patient with cystic fibrosis, and one patient with chronic obstructive pulmonary disease were acquired at 1.5 T. Using a sine model to estimate cardiac and respiratory phases of each image, all images were sorted to reconstruct full cardiac and respiratory cycles. Time to peak (TTP), homogenous ventilation and perfusion maps, and fractional ventilation flow-volume loops were calculated.
RESULTS: For the volunteers, homogenous ventilation and perfusion TTP maps were obtained. The chronic thromboembolic hypertension patient showed increased perfusion TTP in hypoperfused regions in visual agreement with dynamic contrast-enhanced MRI, which improved postpulmonary endaterectomy surgery. Cystic fibrosis and chronic obstructive pulmonary disease patients showed a pattern of increased homogenous ventilation and perfusion TTP in regions of hypoventilation and decreased perfusion. Fractional ventilation flow-volume loops of the chronic obstructive pulmonary disease patient were smaller in comparison with the healthy volunteer, and showed regional differences in visual agreement with functional small airways disease and emphysema on CT.
CONCLUSIONS: This study shows the feasibility of phase-resolved functional lung imaging to gain quantitative information regarding regional lung perfusion and ventilation without the need for ultrafast imaging, which will be advantageous for future clinical translation. Magn Reson Med, 2017. © 2017 International Society for Magnetic Resonance in Medicine.

PMID: 28856715 [PubMed - as supplied by publisher]

Chaperones rescue the energetic landscape of mutant CFTR at single molecule and in cell.

Nat Commun. 2017 Aug 30;8(1):398

Authors: Bagdany M, Veit G, Fukuda R, Avramescu RG, Okiyoneda T, Baaklini I, Singh J, Sovak G, Xu H, Apaja PM, Sattin S, Beitel LK, Roldan A, Colombo G, Balch W, Young JC, Lukacs GL

Abstract
Molecular chaperones are pivotal in folding and degradation of the cellular proteome but their impact on the conformational dynamics of near-native membrane proteins with disease relevance remains unknown. Here we report the effect of chaperone activity on the functional conformation of the temperature-sensitive mutant cystic fibrosis channel (∆F508-CFTR) at the plasma membrane and after reconstitution into phospholipid bilayer. Thermally induced unfolding at 37 °C and concomitant functional inactivation of ∆F508-CFTR are partially suppressed by constitutive activity of Hsc70 and Hsp90 chaperone/co-chaperone at the plasma membrane and post-endoplasmic reticulum compartments in vivo, and at single-molecule level in vitro, indicated by kinetic and thermodynamic remodeling of the mutant gating energetics toward its wild-type counterpart. Thus, molecular chaperones can contribute to functional maintenance of ∆F508-CFTR by reshaping the conformational energetics of its final fold, a mechanism with implication in the regulation of metastable ABC transporters and other plasma membrane proteins activity in health and diseases.The F508 deletion (F508del) in the cystic fibrosis transmembrane conductance regulator (CFTR) is the most common CF causing mutation. Here the authors show that cytosolic chaperones shift the F508del channel conformation to the native fold by kinetic and thermodynamic remodelling of the gating energetics towards that of wild-type CTFR.

PMID: 28855508 [PubMed - in process]

Cystic Fibrosis.

Dtsch Arztebl Int. 2017 Aug 21;114(33-34):564-574

Authors: Naehrig S, Chao CM, Naehrlich L

Abstract
BACKGROUND: Universal screening of newborn babies for cystic fibrosis was launched in Germany on 1 September 2016. Here we present up-to-date information on the diagnosis, treatment, and prognosis of this disease.
METHODS: This article is based on relevant publications retrieved by a selective search in PubMed, along with guidelines from Germany and abroad and systematic reviews.
RESULTS: Cystic fibrosis is caused by a gene mutation leading to dysfunction of the cystic fibrosis transmembrane conductance regulator (CFTR) protein. It affects multiple organ systems-the lungs, pancreas, upper airways, liver, intestine, and reproductive organs-to varying degrees. Its incidence among newborn babies in Germany is between 1 in 3300 and 1 in 4800. Its diagnosis requires both clinical evidence (positive newborn screening, sibling[s] with cystic fibrosis, clinical signs) and the demonstration of CFTR dysfunction by an elevated chloride concentration in sweat, and/or two disease-causing mutations, and/or abnormal electrophysiological findings (nasal potential difference measurement, intestinal short-circuit current measurement). Patients should be cared for by specialized cystic fibrosis centers in close cooperation with their primary care physicians. The median life span of patients with this disease has risen steadily to the current value of 40 years. Aside from symptomatic treatment, the first mutation- specific treatments have recently become available.
CONCLUSION: Early diagnosis and optimized treatment prolong the lives of persons with cystic fibrosis and improve their quality of life. Causally directed treatment for all patients and their effects on the course of disease are now central issues for further research.

PMID: 28855057 [PubMed - in process]

Role of Streptococcus pneumoniae infection in chronic obstructive pulmonary disease patients in Italy.

Ther Adv Respir Dis. 2017 Aug 01;:1753465817728479

Authors: Mantero M, Aliberti S, Azzari C, Moriondo M, Nieddu F, Blasi F, Di Pasquale M

Abstract
BACKGROUND: The aim of this study was to determine the incidence of exacerbations due to Streptococcus pneumoniae in chronic obstructive pulmonary disease (COPD) patients during stable state.
METHODS: We conducted a prospective, observational, cohort study including stable COPD patients, who were evaluated at least every 4 months over a 24-month period at the Respiratory Unit of the IRCCS Policlinico Hospital in Milan, Italy, from 2012 to 2015. Sputum samples were collected at enrollment during stable state to evaluate the frequency of S. pneumoniae colonization and in case of an acute exacerbation to evaluate the incidence of pneumococcal infection.
RESULTS: A total of 79 stable patients with moderate to very severe COPD were enrolled. A total of 217 samples were collected, and 27% ( n = 59) of those were positive for S. pneumoniae. A total of four exacerbations due to S. pneumoniae occurred during follow up (0.31 per 100 person/month). Among positive samples of S. pneumoniae, 109 serotypes were identified. The most frequent serotypes in moderate-to-severe COPD patients during both stable state and exacerbation were 19F (12%), 18 (10%), 19A and 9V (9%) and 35 F (7%). Only 32% of COPD patients were effectively vaccinated for S. pneumoniae with PPV23 vaccine.
CONCLUSION: The most frequent S. pneumoniae serotypes in COPD patients are 19F, 18, 19A, 9V and 35 F, and that almost 50% of S. pneumoniae strains could be covered by PCV13 in adult COPD patients.

PMID: 28854845 [PubMed - as supplied by publisher]

Profile of the ProAxsis Active Neutrophil Elastase immunoassay for precision medicine in chronic respiratory disease.

Expert Rev Mol Diagn. 2017 Aug 31;:

Authors: Keir HR, Fong CJ, Dicker AJ, Chalmers JD

Abstract
INTRODUCTION: Neutrophil elastase (NE) is a 29kDa serine protease released from the azurophilic granules of neutrophils. It may be directly involved in the pathogenesis and disease progression in cystic fibrosis, bronchiectasis and COPD through the degradation of airway elastin and by impairing host defence. Areas covered: Measurement of NE activity has emerged as a promising biomarker strategy in inflammatory lung disease. The authors review studies where NE activity has been linked with clinical outcomes such as lung function decline, exacerbation frequency or other cross-sectional and longitudinal markers of disease severity. In this article the evidence for NE measurement, and the strengths and weaknesses of a commercially available immunoassay which can specifically detect NE activity in human biological samples such as sputum and bronchoalveolar lavage are reviewed. Expert commentary: NE is a promising biomarker for stratifying severity disease. NE also appears to be responsive to antibiotic and other treatments, potentially therefore allowing it to be used as an indicator of treatment response in clinical trials.

PMID: 28854829 [PubMed - as supplied by publisher]