Towards the Development of AgoKirs: New Pharmacological Activators to Study Kir2.x Channel and Target Cardiac Disease.

Int J Mol Sci. 2020 Aug 11;21(16):

Authors: van der Schoor L, van Hattum EJ, de Wilde SM, Harlianto NI, van Weert AJ, Bloothooft M, van der Heyden MAG

Abstract
Inward rectifier potassium ion channels (IK1-channels) of the Kir2.x family are responsible for maintaining a stable negative resting membrane potential in excitable cells, but also play a role in processes of non-excitable tissues, such as bone development. IK1-channel loss-of-function, either congenital or acquired, has been associated with cardiac disease. Currently, basic research and specific treatment are hindered by the absence of specific and efficient Kir2.x channel activators. However, twelve different compounds, including approved drugs, show off-target IK1 activation. Therefore, these compounds contain valuable information towards the development of agonists of Kir channels, AgoKirs. We reviewed the mechanism of IK1 channel activation of these compounds, which can be classified as direct or indirect activators. Subsequently, we examined the most viable starting points for rationalized drug development and possible safety concerns with emphasis on cardiac and skeletal muscle adverse effects of AgoKirs. Finally, the potential value of AgoKirs is discussed in view of the current clinical applications of potentiators and activators in cystic fibrosis therapy.

PMID: 32796537 [PubMed - as supplied by publisher]

Recipient Age Impacts Long-Term Survival in Adult Cystic Fibrosis Subjects after Lung Transplantation.

Ann Am Thorac Soc. 2020 Aug 14;:

Authors: Sethi J, Bugajski A, Patel KN, Davis NM, Wille KM, Qureshi MR, Banday MM, Lin M, Emani V, Weill D, Tumin D, Hayes D, Sharma NS

Abstract
RATIONALE: Lung transplant is an effective treatment option providing survival benefit in patients with cystic fibrosis (CF). Several studies have suggested survival benefit in adults as compared to pediatric CF patients undergoing lung transplant. However, it remains unclear if this age-related disparity persists in adult CF subjects.
OBJECTIVE: We investigated the impact of age at transplant on post-transplant outcomes in adult CF patients.
METHODS: The United Network of Organ Sharing (UNOS) Registry was queried for all adult patients with CF who underwent lung transplantation between 1992 and 2016. Pertinent baseline characteristics, demographics, clinical parameters, and outcomes were recorded. The patients were divided into two groups based on age at transplant: 18- 29 years, and ≥ 30 years. The primary endpoint was survival time. Assessment of post-transplant survival was performed using Kaplan-Meier and log-rank tests with multivariable Cox proportional hazards analysis to adjust for confounding variables.
RESULTS: A total of 3881 CF patients underwent lung transplantation between 1992 and 2016; mean age was 31.0 (± 9.3) years. The 18-29 years old at transplant cohort consisted of 2002 subjects and the ≥30-year-old cohort had 1879 subjects. Survival analysis demonstrated significantly higher survival in subjects in the ≥ 30 years cohort [9.47 years (95% CI; 8.7-10.2)] compared to the 18-29 years cohort [5.21 (95% CI; 4.6-5.8)]. After adjusting for confounders, survival remained higher in recipients aged ≥ 30 years [HR 0.44 (95%CI 0.2 - 0.9)]. Mortality due to allograft failure was significantly lower in CF patients aged ≥ 30 years [28% vs 36.5% OR 0.7 (95% CI 0.6-0.8)], while the incidence of malignancy was higher in the ≥30 years cohort [8% vs 2.9% OR 3.0 (95% CI 1.9-4.6)].
CONCLUSION: Age at transplant influences lung transplant outcomes in CF recipients. CF subjects aged ≥30 years at transplant have superior survival compared to adult CF subjects transplanted between ages 18-29 years.  .

PMID: 32795188 [PubMed - as supplied by publisher]

Fibroblast Growth Factor Receptor 4 Deficiency Mediates Airway Inflammation in the Adult Healthy Lung?

Front Med (Lausanne). 2020;7:317

Authors: Easter M, Garth J, Harris ES, Shei RJ, Helton ES, Wei Y, Denson R, Zaharias R, Rowe SM, Geraghty P, Faul C, Barnes JW, Krick S

Abstract
Fibroblast growth factor receptor (FGFR) 4 has been shown to mediate pro-inflammatory signaling in the liver and airway epithelium in chronic obstructive pulmonary disease. In past reports, FGFR4 knockout (Fgfr4 -/- ) mice did not show any lung phenotype developmentally or at birth, unless FGFR3 deficiency was present simultaneously. Therefore, we wanted to know whether the loss of FGFR4 had any effect on the adult murine lung. Our results indicate that adult Fgfr4 -/- mice demonstrate a lung phenotype consisting of widened airway spaces, increased airway inflammation, bronchial obstruction, and right ventricular hypertrophy consistent with emphysema. Despite downregulation of FGF23 serum levels, interleukin (IL) 1β and IL-6 in the Fgfr4 -/- lung, and abrogation of p38 signaling, primary murine Fgfr4 -/- airway cells showed increased expression of IL-1β and augmented secretion of IL-6, which correlated with decreased airway surface liquid depth as assessed by micro-optical coherence tomography. These findings were paralleled by increased ERK phosphorylation in Fgfr4 -/- airway cells when compared with their control wild-type cells. Analysis of a murine model with constitutive activation of FGFR4 showed attenuation of pro-inflammatory mediators in the lung and airway epithelium. In conclusion, we are the first to show an inflammatory and obstructive airway phenotype in the adult healthy murine Fgfr4 -/- lung, which might be due to the upregulation of ERK phosphorylation in the Fgfr4 -/- airway epithelium.

PMID: 32793609 [PubMed]

Early IL-2 treatment of mice with Pseudomonas aeruginosa pneumonia induced PMN-dominating response and reduced lung pathology.

APMIS. 2020 Aug 13;:

Authors: Calum H, Moser C, Jensen PØ, Bjarnsholt T, Givskov M, Høiby N

Abstract
BACKGROUND: IL-2 is a pro-inflammatory and a Th1 inducing cytokine, which is important for activation of the cell-mediated immunity. IL-2 in serum and sputum has been observed to be reduced in cystic fibrosis (CF) patients. The present IL-2 treatment study of Pseudomonas aeruginosa (PA) lung infected mice was performed in order to evaluate the effect of IL-2 supplement.
METHODS: One hundred-and-twenty female BALB/c mice were divided into three groups: 1) IL-2 treatment/infection (TIG), 2) non-treatment/infection (NTIG), and 3) IL-2 treatment/non-infection (TNIG). The mice were challenged intra-tracheally with PA (PAO579) embedded in seaweed alginate to resemble the biofilm mode of growth. At day 0 and 1, the treatment groups received IL-2 s.c. Mice were killed on day 1 or 2, and cytokine production, lung pathology, and quantitative lung bacteriology was estimated.
RESULTS: IL-2 treatment of infected mice reduced the number of mice with signs of macroscopic lung pathology at day 2 (p<0.05). The reduced macroscopic pathology was paralleled by a reduced IL-1b and TNF-a. In contrast, an increased PMN response at day 2 was observed in the IL-2 treated mice (p<0.01). This was preceded by a significantly higher degree of microscopic inflammation at day 1 (p<0.02). The IL-12 levels decreased in both groups of infected mice at day 2 (p<0.01), however significantly more in the IL-2 treated mice (p<0.02). In accordance, but surprisingly, IFN-g was significantly reduced in the IL-2 treated PA infected group at day 2 (p<0.05).
CONCLUSION: The present results indicate that early IL-2 treatment prolongs the PMN response but also reduces proinflammatory IL-1b and TNF-a and macroscopic signs of pathology.

PMID: 32794206 [PubMed - as supplied by publisher]

CFTR Modulators Dampen Aspergillus-Induced Reactive Oxygen Species Production by Cystic Fibrosis Phagocytes.

Front Cell Infect Microbiol. 2020;10:372

Authors: Currie AJ, Main ET, Wilson HM, Armstrong-James D, Warris A

Abstract
Excessive inflammation by phagocytes during Aspergillus fumigatus infection is thought to promote lung function decline in CF patients. CFTR modulators have been shown to reduce A. fumigatus colonization in vivo, however, their antifungal and anti-inflammatory mechanisms are unclear. Other treatments including azithromycin and acebilustat may dampen Aspergillus-induced inflammation due to their immunomodulatory properties. Therefore, we set out in this study to determine the effects of current CF therapies on ROS production and fungal killing, either direct or indirect by enhancing antifungal immune mechanisms in peripheral blood immune cells from CF patients upon A. fumigatus infection. Isolated peripheral blood mononuclear cells (PBMCs) and polymorphonuclear cells (PMNs) from CF patients and healthy volunteers were challenged with A. fumigatus following pre-treatment with CFTR modulators, azithromycin or acebilustat. Ivacaftor/lumacaftor treated CF and control subject PMNs resulted in a significant reduction (p < 0.05) in Aspergillus-induced ROS. For CF PBMC, Aspergillus-induced ROS was significantly reduced when pre-treated with ivacaftor alone (p < 0.01) or in combination with lumacaftor (p < 0.01), with a comparable significant reduction in control subject PBMC (p < 0.05). Azithromycin and acebilustat had no effect on ROS production by CF or control subject phagocytes. None of the treatments showed an indirect or direct antifungal activity. In summary, CFTR modulators have potential for additional immunomodulatory benefits to prevent or treat Aspergillus-induced inflammation in CF. The comparable effects of CFTR modulators observed in phagocytes from control subjects questions their exact mechanism of action.

PMID: 32793514 [PubMed - in process]

Using registries for research in CF. How can we be sure about the outputs?

J Cyst Fibros. 2019 05;18(3):309-310

Authors: Elborn JS, Gonska T

PMID: 30982754 [PubMed - indexed for MEDLINE]

Gene Therapy in Rare Respiratory Diseases: What Have We Learned So Far?

J Clin Med. 2020 Aug 08;9(8):

Authors: Bañuls L, Pellicer D, Castillo S, Navarro-García MM, Magallón M, González C, Dasí F

Abstract
Gene therapy is an alternative therapy in many respiratory diseases with genetic origin and currently without curative treatment. After five decades of progress, many different vectors and gene editing tools for genetic engineering are now available. However, we are still a long way from achieving a safe and efficient approach to gene therapy application in clinical practice. Here, we review three of the most common rare respiratory conditions-cystic fibrosis (CF), alpha-1 antitrypsin deficiency (AATD), and primary ciliary dyskinesia (PCD)-alongside attempts to develop genetic treatment for these diseases. Since the 1990s, gene augmentation therapy has been applied in multiple clinical trials targeting CF and AATD, especially using adeno-associated viral vectors, resulting in a good safety profile but with low efficacy in protein expression. Other strategies, such as non-viral vectors and more recently gene editing tools, have also been used to address these diseases in pre-clinical studies. The first gene therapy approach in PCD was in 2009 when a lentiviral transduction was performed to restore gene expression in vitro; since then, transcription activator-like effector nucleases (TALEN) technology has also been applied in primary cell culture. Gene therapy is an encouraging alternative treatment for these respiratory diseases; however, more research is needed to ensure treatment safety and efficacy.

PMID: 32784514 [PubMed]

Cystic Fibrosis-Screening Positive Inconclusive Diagnosis: Newborn Screening and Long-Term Follow-Up Permits to Early Identify Patients with CFTR-Related Disorders.

Diagnostics (Basel). 2020 Aug 08;10(8):

Authors: Castaldo A, Cimbalo C, Castaldo RJ, D'Antonio M, Scorza M, Salvadori L, Sepe A, Raia V, Tosco A

Abstract
BACKGROUND: Newborn screening (NBS) early-identifies cystic fibrosis (CF), but in CF-screening positive inconclusive diagnosis (CF-SPID) the results of immunoreactive trypsinogen (IRT), molecular analysis and sweat test (ST) are discordant. A percentage of CF-SPID evolves to CF, but data on long-term monitoring are lacking. We describe the follow-up of all CF and CF-SPID identified between 2008 and 2019.
METHODS: NBS was performed by IRT followed by molecular analysis and ST between 2008 and 2014; double IRT followed by molecular analysis and ST after 2014.
RESULTS: NBS revealed 47 CF and 99 CF-SPID newborn, a ratio 1:2.1-the highest reported so far. This depends on the identification by gene sequencing of the second variant with undefined effect in 40 CF-SPID that otherwise would have been defined as carriers. Clinical complications and pulmonary infections occurred more frequently among CF patients than among CF-SPID. Two CF-SPID cases evolved to CF (at two years), while eight evolved to CFTR-related disorders (CFTR-RD), between one and eight years, with bronchiectasis (two), recurrent pneumonia (four, two with sinonasal complications), recurrent pancreatitis (two). No clinical, biochemical or imaging data predicted the evolution.
CONCLUSION: Gene sequencing within the NBS reveals a higher number of CF-SPID and we first describe an approach to early identify CFTR-RD, with relevant impact on their outcome.

PMID: 32784480 [PubMed]

Non-viral mediated gene therapy in human cystic fibrosis airway epithelial cells recovers chloride channel functionality.

Int J Pharm. 2020 Aug 10;:119757

Authors: Sainz-Ramos M, Villate-Beitia I, Gallego I, Qtaish NAL, Lopez-Mendez TB, Eritja R, Grijalvo S, Puras G, Pedraz JL

Abstract
Gene therapy strategies based on non-viral vectors are currently considered as a promising therapeutic option for the treatment of cystic fibrosis (CF), being liposomes the most commonly used gene carriers. Niosomes offer a powerful alternative to liposomes due to their higher stability and lower cytotoxicity, provided by their non-ionic surfactant and helper components. In this work, a three-formulation screening ois performed, in terms of physicochemical and biological behavior, in CF patient derived airway epithelial cells. The most efficient niosome formulation reaches 28% of EGFP expressing live cells and follows caveolae-mediated endocytosis. Transfection with therapeutic cystic fibrosis transmembrane conductance regulator (CFTR) gene results in 5-fold increase of CFTR protein expression in transfected versus non-transfected cells, which leads to 1.5-fold increment of the chloride channel functionality. These findings highlight the relevance of niosome-based systems as an encouraging non-viral gene therapy platform with potential therapeutic benefits for CF.

PMID: 32791297 [PubMed - as supplied by publisher]

Delafloxacin - A Novel fluoroquinolone for the treatment of ciprofloxacin-resistant pseudomonas aeruginosa in patients with cystic fibrosis (CF).

Clin Respir J. 2020 Aug 13;:

Authors: Millar BC, McCaughan J, Rendall JC, Moore JE

Abstract
INTRODUCTION: Fluoroquinolone antibiotics, namely ciprofloxacin and levofloxacin, play an important role in treating infection in cystic fibrosis (CF) and ciprofloxacin remains the last orally available antipseudomonal agent. Recently, a new fluoroquinolone, delafloxacin, has been approved by the FDA for skin infections. This antibiotic is a novel dual-targeting anionic fluoroquinolone and differs from previous agents in its class, as it lacks a protonatable substituent. To date, there are no reports of its use or activity against Pseudomonas aeruginosa (PA) in CF. Alarmingly, fluoroquinolone resistance is increasing amongst CF PA isolates. The aims of the study were to (i) examine in vitro susceptibility of delafloxacin against a population of PA (n=52) isolated from adult CF patients at our CF centre, (ii) to compare delafloxacin and ciprofloxacin susceptibilities and (iii) to evaluate where delafloxacin may add benefit in treating CF PA.
METHODS: In vitro susceptibilities were examined on 52 non-mucoid PA and ATCC™ 27853 reference strain, by employing Etest® gradient for delafloxacin (range:0.002 32 mg/L) and ciprofloxacin (0.002-32 mg/L), as per manufacturer's instructions (Biomerieux).
RESULTS: MIC range, MIC50 and MIC90 for delafloxacin were 0.064->32 mg/L, 0.56 mg/L and 2.19 mg/L, respectively. For ciprofloxacin, these were 0.047->32 mg/L, 1.69 mg/L and 8.0 mg/L, respectively. Overall, isolates were statistically more sensitive to delafloxacin (p=0.0005) than ciprofloxacin. Of note, 4/12 (33.3%) isolates with intermediate resistance to ciprofloxacin were sensitive to delafloxacin. Similarly, 10/28 (35.7%) isolates resistant to ciprofloxacin were sensitive to delafloxacin, with only 17.9% isolates resistant to ciprofloxacin, resistant to delafloxacin CONCLUSION: Given similar breakpoints of these fluoroquinolones, this data shows that delafloxacin has greater activity than ciprofloxacin. Whilst delafloxacin and ciprofloxacin were equally effective with sensitive isolates, the value of delafloxacin was noted with more resistant isolates to ciprofloxacin. Whilst ciprofloxacin should remain the first line fluoroquinolone for treating CF PA, delafloxacin shows potential in treating ciprofloxacin-resistant PA.

PMID: 32790958 [PubMed - as supplied by publisher]

Sensing of electrolytes in urine using a miniaturized paper-based device.

Sci Rep. 2020 Aug 12;10(1):13620

Authors: Ghaderinezhad F, Ceylan Koydemir H, Tseng D, Karinca D, Liang K, Ozcan A, Tasoglu S

Abstract
Analyzing electrolytes in urine, such as sodium, potassium, calcium, chloride, and nitrite, has significant diagnostic value in detecting various conditions, such as kidney disorder, urinary stone disease, urinary tract infection, and cystic fibrosis. Ideally, by regularly monitoring these ions with the convenience of dipsticks and portable tools, such as cellphones, informed decision making is possible to control the consumption of these ions. Here, we report a paper-based sensor for measuring the concentration of sodium, potassium, calcium, chloride, and nitrite in urine, accurately quantified using a smartphone-enabled platform. By testing the device with both Tris buffer and artificial urine containing a wide range of electrolyte concentrations, we demonstrate that the proposed device can be used for detecting potassium, calcium, chloride, and nitrite within the whole physiological range of concentrations, and for binary quantification of sodium concentration.

PMID: 32788641 [PubMed - as supplied by publisher]

Paediatric reproducibility limits for the forced expiratory volume in 1 s.

Thorax. 2020 Aug 11;:

Authors: Stanojevic S, Filipow N, Ratjen F

Abstract
BACKGROUND: Current reproducibility standards for spirometry were derived using a small adult dataset and may not be optimal for interpretation of repeated measurements of lung function in children.
OBJECTIVE: To define reproducibility limits for forced expiratory volume in 1 s (FEV1) change that represent the normal within-subject between-visit variability in healthy children and evaluate these limits as a tool to monitor children with cystic fibrosis (CF).
METHODS: Repeated FEV1 measurements (3 months to 5 years apart) from healthy children from the Global Lung Function Initiative data repository were used to derive a conditional change score. Spirometry and clinical data from a CF clinical database was used to verify utility in clinical practice.
RESULTS: A reproducibility change score was derived from 47 938 FEV1 measures from 7885 healthy children 6-18 years of age. The simple algorithm, which is conditional on the initial measurement, also accounts for age and time interval between measurements. The change score limits of reproducibility were much narrower than currently used cut-offs. Specifically, changes, considered as improvements using either a 12% or 10% relative change from baseline, are too wide for children. In CF, there was overall agreement between different approaches, with the distinct advantage that the change score was not biased by regression to the mean.
CONCLUSIONS: Compared with current approaches to interpretation of repeated lung function measurements, the proposed change score was less biased and provides a simple alternative to reduce misinterpretation.

PMID: 32788261 [PubMed - as supplied by publisher]

FUNCTIONAL PERFORMANCE IN THE MODIFIED SHUTTLE TEST IN CHILDREN AND ADOLESCENTS WITH CYSTIC FIBROSIS.

Rev Paul Pediatr. 2021;39:e2019322

Authors: Leite LR, Queiroz KCV, Coelho CC, Vergara AA, Donadio MVF, Aquino EDS

Abstract
OBJECTIVE: To evaluate factors associated with the performance of children and adolescents with cystic fibrosis (CF) in the Modified Shuttle Test (MST) and compare it with healthy children and adolescents.
METHODS: This is a cross-sectional study, with children and adolescents divided into two groups: cystic fibrosis (CFG) and control (CG). Variables evaluated in the MST: walking distance, test level, heart rate variation (∆Hr), post-test mean arterial pressure (MAP Pt) and peripheral oxygen saturation variation (∆SPO2). Statistical analysis included Mann Whitney and Spearman coefficient tests, being significant p<0.05.
RESULTS: Sixty individuals aged 6-16 years old were evaluated. Anthropometric data was similar between groups. Differences between groups were shown for: baseline heart rate (BHr), peak heart rate (PHr), ∆Hr, recovery heart rate (RHr), post-test respiratory rate (PtBr), saturation variables, peripheral oxygen level (SpO2B) and level test. The ∆Hr and MAP Pt had a moderate positive correlation with distance and level test for both groups (respectively: r=0.6 / p<0.001; r=0.6 / p<0.001). In CFG, the level test had a significant association (r=0.4 - p=0.02) with %FEV1.
CONCLUSIONS: Children with cystic fibrosis presented functional limitation in the Modified Shuttle Test, which was influenced by lung function.

PMID: 32785464 [PubMed - in process]

Synthesis and biological evaluation of thiazole derivatives on basic defects underlying cystic fibrosis.

Bioorg Med Chem Lett. 2020 Aug 09;:127473

Authors: Pesce E, Pedemonte N, Leoni A, Locatelli A, Morigi R

Abstract
Cystic fibrosis is a genetic disease caused by loss-of-function mutations in the cystic fibrosis transmembrane conductance regulator gene, encoding for CFTR protein. The most frequent mutation is the deletion of phenylalanine at position 508 (F508del), which leads to distinct defects in channel gating and cellular processing. In last years, several thiazole containing small molecules, endowed with dual F508del-CFTR modulator activity, proved to be able to target these defects. In search of new chemical entities able to restore CFTR function, we designed and synthesized a small series of sixteen thiazole derivatives. The designed compounds were studied as correctors and potentiators of F508del-CFTR. Although none of the molecules showed significant corrector activity, compounds 10 and 11 exhibited potentiator effects, thus allowing to determine some basic structural features which enable to obtain F508del-CFTR potentiator activity. In silico ADME studies showed that these derivatives obey Lipinski's rule of five and are expected to be orally bioavailable. Therefore, these molecules may represent a good starting point for the design of analogues endowed with improved CFTR potentiator activity and a good pharmacokinetic profile.

PMID: 32784089 [PubMed - as supplied by publisher]

Stem Cells and Lung Regeneration.

Am J Physiol Cell Physiol. 2020 Aug 12;:

Authors: Parekh KR, Nawroth J, Pai A, Busch SM, Senger CN, Ryan AL

Abstract
Summary/Abstract The ability to replace defective cells in an airway with cells that can engraft, integrate and restore a functional epithelium could potentially cure a number of lung diseases. Progress toward the development of strategies to regenerate the adult lung by either in vivo or ex vivo targeting of endogenous stem cells or pluripotent stem cell derivatives, is limited by our fundamental lack of understanding of the mechanisms controlling human lung development, the precise identity and function of human lung stem and progenitor cell types, and the genetic and epigenetic control of human lung fate. In this review, we intend to discuss the known stem/progenitor cell populations, their relative differences between rodents and humans, their roles in chronic lung disease, and their therapeutic prospects. Additionally, we highlight the recent breakthroughs that have increased our understanding of these cell types. These advancements include novel lineage-traced animal models and single-cell RNA sequencing of human airway cells, which have provided critical information on the stem cell subtypes, transition states, identifying cell markers, and intricate pathways that commit a stem cell to differentiate or to maintain plasticity. As our capacity to model the human lung evolves, so will our understanding of lung regeneration and our ability to target endogenous stem cells as a therapeutic approach for lung disease.

PMID: 32783658 [PubMed - as supplied by publisher]

Quantification of bronchoalveolar neutrophil extracellular traps and phagocytosis in murine pneumonia.

Am J Physiol Lung Cell Mol Physiol. 2020 Aug 12;:

Authors: Gautam S, Stahl Y, Young GM, Howell R, Cohen AJ, Tsang DA, Martin T, Sharma L, Dela Cruz CS

Abstract
The past two decades have witnessed a resurgence in neutrophil research, inspired in part by the discovery of neutrophil extracellular traps (NETs) and their myriad roles in health and disease. Within the lung, dysregulation of neutrophils and NETosis have been linked to an array of diseases including pneumonia, cystic fibrosis, acute respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD), and severe asthma. However, our understanding of pathologic neutrophil responses in the lung remains incomplete. Two methodologic issues have contributed to this gap: first, an emphasis on studying neutrophils from blood rather than the lung; and second, the technical difficulties of interrogating neutrophil responses in mice, which has largely restricted basic murine research to specialized laboratories. To address these limitations, we have developed a suite of techniques for studying neutrophil effector functions specifically in the mouse lung. These include ex vivo assays for phagocytosis and NETosis using bronchoalveolar neutrophils, and in situ evaluation of NETosis in a murine model of pneumonia. Throughout, we have prioritized technical ease and robust, quantitative readouts. We hope these assays will help to standardize research on lung neutrophils and improve accessibility to this burgeoning field.

PMID: 32783617 [PubMed - as supplied by publisher]

First contact: The role of respiratory cilia in host-pathogen interactions in the airways.

Am J Physiol Lung Cell Mol Physiol. 2020 Aug 12;:

Authors: Kuek LE, Lee RJ

Abstract
Respiratory cilia are the driving force of the mucociliary escalator, working in conjunction with secreted airway mucus to clear inhaled debris and pathogens from the conducting airways. Respiratory cilia are also one of the first contact points between host and inhaled pathogens. Impaired ciliary function is a common pathological feature in patients with chronic airway diseases, increasing susceptibility to respiratory infections. Common respiratory pathogens including viruses, bacteria and fungi have been shown to target cilia and/or ciliated airway epithelial cells, resulting in a disruption of mucociliary clearance that may facilitate host infection. Despite being an integral component of airway innate immunity, the role of respiratory cilia and their clinical significance during airway infections is still poorly understood. This review examines the expression, structure, and function of respiratory cilia during pathogenic infection of the airways. This review also discusses specific known points of interaction of bacteria, fungi, and viruses with respiratory cilia function. The emerging biological functions of motile cilia relating to intracellular signaling and their potential immuno-regulatory roles during infection will also be discussed.

PMID: 32783615 [PubMed - as supplied by publisher]

Foreign body and caustic ingestions in children: A clinical practice guideline.

Dig Liver Dis. 2020 Aug 08;:

Authors: Oliva S, Romano C, De Angelis P, Isoldi S, Mantegazza C, Felici E, Dabizzi E, Fava G, Renzo S, Strisciuglio C, Quitadamo P, Saccomani MD, Bramuzzo M, Orizio P, Nardo GD, Bortoluzzi F, Pellegrino M, Illiceto MT, Torroni F, Cisarò F, Zullo A, Macchini F, Gaiani F, Raffaele A, Bizzarri B, Arrigo S, De' Angelis GL, Martinelli M, Norsa L, Italian Society of Pediatric Gastroenterology Hepatology and Nutrition (SIGENP), and The Italian Association of Hospital Gastroenterologists and Endoscopists (AIGO)

Abstract
Foreign body and caustic ingestions in children are usually the most common clinical challenges for emergency physicians, general pediatricians and pediatric gastroenterologists. Management of these conditions often requires different levels of expertise and competence. Endoscopy is often necessary but there is a high risk of misusing this tool with incorrect timing and indications. The imprecise clinical history frequently leaves clinicians uncertain about timing and nature of the ingestion. Few clinical guidelines regarding management of these ingestions in children have been published, none of which from the Italian Society of Pediatric Gastroenterology, Hepatology and Nutrition (SIGENP). An expert panel of Italian endoscopists was convened by the SIGENP Endoscopy Working Group to produce the present article that outlines practical clinical approaches to the pediatric patient with a variety of foreign body and caustic ingestions. The Italian Association of Hospital Gastroenterologists and Endoscopists (AIGO) has also endorsed the project since many adult endoscopists usually manage children with these conditions. Differently from the other published guidelines, the proposed one focuses on the role of the endoscopists (regardless of whether they are adult or pediatric gastroenterologists) in the diagnostic process of children with foreign body and caustic ingestions.

PMID: 32782094 [PubMed - as supplied by publisher]

Factoring in the Complexity of the Cystic Fibrosis Lung to Understand Aspergillus fumigatus and Pseudomonas aeruginosa Interactions.

Pathogens. 2020 Aug 06;9(8):

Authors: Beswick E, Amich J, Gago S

Abstract
Pseudomonas aeruginosa has long been established as the most prevalent respiratory pathogen in Cystic Fibrosis (CF) patients, with opportunistic infection causing profound morbidity and mortality. Recently, Aspergillus fumigatus has also been recognised as a key contributor to CF lung deterioration, being consistently associated with decreased lung function and worsened prognosis in these patients. As clinical evidence for the common occurrence of combined infection with these two pathogens increases, research into the mechanism and consequences of their interaction is becoming more relevant. Clinical evidence suggests a synergistic effect of combined infection, which translates into a poorer prognosis for the patients. In vitro results from the laboratory have identified a variety of possible synergistic and antagonistic interactions between A. fumigatus and P. aeruginosa. Here, we present a comprehensive overview of the complex environment of the CF lung and discuss how it needs to be considered to determine the exact molecular interactions that A. fumigatus and P. aeruginosa undergo during combined infection and their effects on the host.

PMID: 32781694 [PubMed]

Cystic Fibrosis Defective Response to Infection Involves Autophagy and Lipid Metabolism.

Cells. 2020 Aug 06;9(8):

Authors: Mingione A, Ottaviano E, Barcella M, Merelli I, Rosso L, Armeni T, Cirilli N, Ghidoni R, Borghi E, Signorelli P

Abstract
Cystic fibrosis (CF) is a hereditary disease, with 70% of patients developing a proteinopathy related to the deletion of phenylalanine 508. CF is associated with multiple organ dysfunction, chronic inflammation, and recurrent lung infections. CF is characterized by defective autophagy, lipid metabolism, and immune response. Intracellular lipid accumulation favors microbial infection, and autophagy deficiency impairs internalized pathogen clearance. Myriocin, an inhibitor of sphingolipid synthesis, significantly reduces inflammation, promotes microbial clearance in the lungs, and induces autophagy and lipid oxidation. RNA-seq was performed in Aspergillusfumigatus-infected and myriocin-treated CF patients' derived monocytes and in a CF bronchial epithelial cell line. Fungal clearance was also evaluated in CF monocytes. Myriocin enhanced CF patients' monocytes killing of A. fumigatus. CF patients' monocytes and cell line responded to infection with a profound transcriptional change; myriocin regulates genes that are involved in inflammation, autophagy, lipid storage, and metabolism, including histones and heat shock proteins whose activity is related to the response to infection. We conclude that the regulation of sphingolipid synthesis induces a metabolism drift by promoting autophagy and lipid consumption. This process is driven by a transcriptional program that corrects part of the differences between CF and control samples, therefore ameliorating the infection response and pathogen clearance in the CF cell line and in CF peripheral blood monocytes.

PMID: 32781626 [PubMed - in process]