A chronic strain of the cystic fibrosis pathogen Pandoraea pulmonicola expresses a heterogenous hypo-acylated lipid A.

Glycoconj J. 2020 Oct 13;:

Authors: Pither MD, McClean S, Silipo A, Molinaro A, Di Lorenzo F

Abstract
Pandoraea sp. is an emerging Gram-negative pathogen in cystic fibrosis causing severe and persistent inflammation and damage of the lungs. The molecular mechanisms underlying the high pathogenicity of Pandoraea species are still largely unknown. As Gram-negatives, Pandoraea sp. express lipopolysaccharides (LPS) whose recognition by the host immune system triggers an inflammatory response aimed at the bacterial eradication from the infected tissues. The degree of the inflammatory response strongly relies on the fine structure of the LPS and, in particular, of its glycolipid moiety, i.e. the lipid A. Here we report the structure of the lipid A isolated from the LPS of a chronic strain of P. pulmonicola (RL 8228), one of the most virulent identified so far among the Pandoraea species. Our data demonstrated that the examined chronic strain produces a smooth-type LPS with a complex mixture of hypoacylated lipid A species displaying, among other uncommon characteristics, the 2-hydroxylation of some of the acyl chains and the substitution by an additional glucosamine on one or both the phosphate groups.

PMID: 33048281 [PubMed - as supplied by publisher]

Extended-criteria uncontrolled DCD donor for a fragile recipient: A case report about a challenging yet successful lung transplantation.

Int J Surg Case Rep. 2020 Sep 14;:

Authors: Musso V, Mendogni P, Scaravilli V, Morlacchi LC, Croci GA, Palleschi A

Abstract
INTRODUCTION: Lung donation after circulatory death (DCD) has proved to be an effective strategy for expanding the donor pool, but is still considered challenging. We report a successful case of lung procurement from an extended-criteria uncontrolled DCD.
PRESENTATION OF CASE: We evaluated the lungs of an uncontrolled DCD from a hospital without extracorporeal membrane oxygenation (ECMO) program. The donor was a non-smoker 20-year old male with a history of cardiomyopathy, cardiocirculatory arrests, and Lennox-Gastaut syndrome. Cardiac arrest occurred in a swimming pool, and bronchoscopy showed signs of inhalation. We employed our usual normothermic in-situ open-ventilated lung approach. After retrieval, lungs were stored on ice, then evaluated with ex-vivo lung perfusion (EVLP) and judged suitable for transplantation. The recipient was a 26-year old female with cystic fibrosis on long-term oxygen therapy, on the waitlist for up to 21 months due to her anthropomorphic characteristics. She required central VA-ECMO support during bilateral lung transplantation. Primary graft dysfunction (PGD) within the first 72 h reached grade 3; post-operative peripheral VV-ECMO support was discontinued two days after surgery. The patient was discharged 28 days after surgery; she is alive two years after transplantation with no signs of rejection nor anastomotic complications.
DISCUSSION: Despite the spreading use of lungs from controlled DCD, perplexities remain on uncontrolled DCD, namely: severe PDG, postoperative mortality, airway complications.
CONCLUSION: Our case report suggests that good results can be achieved with uncontrolled DCD despite the presence of relative contraindications: inhalation of water, prolonged ischemic times and recipient in poor conditions.

PMID: 33046417 [PubMed - as supplied by publisher]

[Neonatal cystic fibrosis screening: Analysis and differences in immunoreactive trypsin levels in newborns with a positive screen].

An Pediatr (Barc). 2020 Oct 09;:

Authors: Arrudi-Moreno M, García-Romero R, Samper-Villagrasa P, Sánchez-Malo MJ, Martin-de-Vicente C

Abstract
INTRODUCTION: Neonatal cystic fibrosis (CF) screening has enabled the disease to be diagnosed early, and is a determining factor in the increase in survival of these patients. Its main disadvantage is its low specificity and elevated number of false positives. The aim of this study is to analyse the differences in immunoreactive trypsin (IRT) between the different groups of newborns (NB) with a positive neonatal screen depending on whether they were healthy, healthy carriers, affected by CF, or CFSPID (Cystic Fibrosis Screen Positive, Inconclusive Diagnosis).
MATERIAL: Retrospective analytical study of the concentrations of IRT in NB with a positive neonatal screen for CF born in a tertiary hospital during an 8-year period RESULTS: A total of 790 NB with a positive neonatal screen for CF were analysed. Of these 86.3% were term, 53% girls, and 11.8% were admitted. The mean IRT value was 79.16 ng/mL (range 60 - 367). Significantly higher concentrations of IRT were found in those affected by CF compared to the other groups (P<.001). There were higher levels in large prematures (P=.007) and admitted patients (P=.002). There were no differences as regards gender or season. There was a direct correlation of 64% (P=.001) between IRT and sweat test in those affected by CF and CFSPID. A cut-off value of IRT for the diagnosis of CF was calculated from the ROC curve (76.2 ng/mL (S=95.7%, Sp=64.5%).
CONCLUSIONS: NB with CF have significantly higher levels of IRT than healthy ones, or carriers and CFSPID. Prematurity and hospital admission may also have an influence. A higher IRT value is associated with a higher level in the sweat test.

PMID: 33046410 [PubMed - as supplied by publisher]

CRISPR/Cas9 genome editing tool: A Promising Tool for Therapeutic Applications on respiratory diseases.

Curr Gene Ther. 2020 Oct 12;:

Authors: Shaikh SB, Bhandary YP

Abstract
Respiratory diseases are one of the prime topics of concern in the current era due to improper diagnostics tools. Gene-editing therapy like Clustered regularly interspaced palindromic repeats-associated nuclease 9 (CRISPR/Cas9) is gaining popularity in pulmonary research opening up doors to invaluable insights on underlying mechanisms. CRISPR/Cas9 can be considered as a potential gene editing tool with a scientific community that is helping in the advancement of knowledge in respiratory health and therapy. As an appealing therapeutic tool, we hereby explore the advanced research on the application of CRISPR/Cas9 tool in chronic respiratory diseases such as lung cancer, Acute respiratory distress syndrome (ARDS), cystic fibrosis (CF). We also address the urgent need to establish this gene-editing tool in various other lung diseases such as asthma, Chronic obstructive pulmonary disease (COPD) and Idiopathic pulmonary fibrosis (IPF). The present review introduces CRISPR/Cas9 as a worthy application in targeting epithelial-mesenchymal transition and fibrinolytic system via editing specific genes. Thereby, based on the efficiency of CRISPR/Cas9, it can be considered as a promising therapeutic tool in respiratory health research.

PMID: 33045965 [PubMed - as supplied by publisher]

Six-Minute Walk Test: Clinical Role, Technique, Coding, and Reimbursement.

Chest. 2020 03;157(3):603-611

Authors: Agarwala P, Salzman SH

Abstract
The 6-min walk test (6MWT) is a commonly used test for the objective assessment of functional exercise capacity for the management of patients with moderate-to-severe pulmonary disease. Unlike pulmonary function testing, the 6MWT captures the often coexisting extrapulmonary manifestations of chronic respiratory disease, including cardiovascular disease, frailty, sarcopenia, and cancer. In contrast with cardiopulmonary exercise stress testing, this test does not require complex equipment or technical expertise. In this low complexity, safe test, the patient is asked to walk as far as possible along a 30-m minimally trafficked corridor for a period of 6 min with the primary outcome measure being the 6-min walk distance (6MWD) measured in meters. There has been interest in other derived indexes, such as distance-desaturation product (the product of nadir oxygen saturation and walk distance), which in small studies has been predictive of morbidity and mortality in certain chronic respiratory conditions. Special attention to methodology is required to produce reliable and reproducible results. Factors that can affect walk distance include track layout (continuous vs straight), track length, oxygen amount and portability, learning effect, and verbal encouragement. The absolute 6MWD and change in 6MWD are predictive of morbidity and mortality in patients with COPD, pulmonary arterial hypertension, and idiopathic pulmonary fibrosis and patients awaiting lung transplant, highlighting its use in management decisions and clinical trials. As of January 2018, Current Procedural Terminology code 94620 (simple pulmonary stress test) has been deleted and replaced by two new codes, 94617 and 94618. Code 94617 includes exercise test for bronchospasm including pre- and postspirometry, ECG recordings, and pulse oximetry. Code 94618, pulmonary stress testing (eg, 6MWT), includes the measurement of heart rate, oximetry, and oxygen titration when performed. If 94620 is billed after January 2018 it will not be reimbursed.

PMID: 31689414 [PubMed - indexed for MEDLINE]

Transcript from Stuart Elborn's address at the ECFC award in June 2019.

J Cyst Fibros. 2019 09;18(5):587-589

Authors:

PMID: 31500811 [PubMed - indexed for MEDLINE]

Important steps in the journey to highly effective CFTR modulator access for people with CF.

J Cyst Fibros. 2019 09;18(5):577-578

Authors: VanDevanter DR, Mayer-Hamblett N

PMID: 31500809 [PubMed - indexed for MEDLINE]

Fair selection of participants in clinical trials: The challenge to push the envelope further.

J Cyst Fibros. 2019 09;18(5):e48-e50

Authors: Davies JC, Scott S, Dobra R, Brendell R, Brownlee K, Carr SB, Cosgriff R, Simmonds NJ, London Network of Clinical Trials Accelerator Platform sites, Jahan R, Jones A, Matthews J, Brown S, Galono K, Miles K, Pao C, Shafi N, Watson D, Orchard C, Davies G, Pike K, Shah S, Bossley CJ, Fong T, Macedo P, Ruiz G, Waller M, Baker L

PMID: 31405729 [PubMed - indexed for MEDLINE]

Theranostics by testing CFTR modulators in patient-derived materials: The current status and a proposal for subjects with rare CFTR mutations.

J Cyst Fibros. 2019 09;18(5):685-692

Authors: Amaral MD, de Boeck K, ECFS Strategic Planning Task Force on ‘Speeding up access to new drugs for CF’

Abstract
The last decade has witnessed developments in the CF drug pipeline which are both exciting and unprecedented, bringing with them previously unconsidered challenges. The Task Force group came together to consider these challenges and possible strategies to address them. Over the last 18 months, we have discussed internally and gathered views from a broad range of individuals representing patient organizations, clinical and research teams, the pharmaceutical industry and regulatory agencies. In this and the accompanying article, we discuss two main areas of focus: i) optimising trial design and delivery for speed/efficiency; ii) drug development for patients with rare CFTR mutations. We propose some strategies to tackle the challenges ahead and highlight areas where further thought is needed. We see this as the start of a process rather than the end and hope herewith to engage the wider community in seeking solutions to improved treatments for all patients with CF.

PMID: 31326274 [PubMed - indexed for MEDLINE]

Decreased mRNA and protein stability of W1282X limits response to modulator therapy.

J Cyst Fibros. 2019 09;18(5):606-613

Authors: Aksit MA, Bowling AD, Evans TA, Joynt AT, Osorio D, Patel S, West N, Merlo C, Sosnay PR, Cutting GR, Sharma N

Abstract
BACKGROUND: Cell-based studies have shown that W1282X generates a truncated protein that can be functionally augmented by modulators. However, modulator treatment of primary cells from individuals who carry two copies of W1282X generates no functional CFTR. To understand the lack of response to modulators, we investigated the effect of W1282X on CFTR RNA transcript levels.
METHODS: qRT-PCR and RNA-seq were performed on primary nasal epithelial (NE) cells of a previously studied individual who is homozygous for W1282X, her carrier parents and control individuals without nonsense variants in CFTR.
RESULTS: CFTR RNA bearing W1282X in NE cells shows a steady-state level of 4.2 ± 0.9% of wild-type (WT) CFTR RNA in the mother and 12.4 ± 1.3% in the father. NMDI14, an inhibitor of nonsense-mediated mRNA decay (NMD), restored W1282X mRNA to almost 50% of WT levels in the parental NE cells. RNA-seq of the NE cells homozygous for W1282X showed that CFTR transcript level was reduced to 1.7% of WT (p-value: 4.6e-3). Negligible truncated CFTR protein was generated by Flp-In 293 cells stably expressing the W1282X EMG even though CFTR transcript was well above levels observed in the parents and proband. Finally, we demonstrated that NMD inhibition improved the stability and response to correctors of W1282X-CFTR protein expressed in the Flp-In-293 cells.
CONCLUSION: These results show that W1282X can cause substantial degradation of CFTR mRNA that has to be addressed before efforts aimed at augmenting CFTR protein function can be effective.

PMID: 30803905 [PubMed - indexed for MEDLINE]

A systematic review on the impact of appropriate versus inappropriate initial antibiotic therapy on the outcomes of patients with severe bacterial infections.

Int J Antimicrob Agents. 2020 Oct 09;:106184

Authors: Bassetti M, Rello J, Blasi F, Goossens H, Sotgiu G, Tavoschi L, Zasowski EJ, Arber MR, McCool R, Patterson JV, Longshaw CM, Lopes S, Manissero D, Nguyen ST, Tone K, Aliberti S

Abstract
BACKGROUND: We investigated the impact of appropriate versus inappropriate initial antimicrobial therapy on the clinical outcomes of patients with severe bacterial infections as part of a systematic review with meta-analyses assessing the impact of delay in appropriate antimicrobial therapy.
METHODS: Literature searches of MEDLINE and Embase, conducted on 24 July 2018, identified studies published after 2007 reporting the impact of delay in appropriate antibiotic therapy for hospitalised adult patients with bacterial infections. Results were statistically pooled for outcomes including mortality, length of hospital stay (LOS) and treatment failure. Subgroup analyses were explored by site of infection where data permitted.
RESULTS: Inclusion criteria were met by 145 studies, of which 122 reported data on the impact of appropriate versus inappropriate initial therapy. In pooled analysis, rates of mortality were significantly in favour of appropriate therapy (odds ratio [OR] 0.44 [95% CI 0.39-0.50]). Across ten studies, LOS was significantly shorter with appropriate therapy compared with inappropriate therapy (mean difference [MD] -2.95 days [95% CI -5.46 to -0.43]). In patients who received appropriate therapy, incidence of treatment failure was significantly lower compared with patients who received inappropriate therapy (six studies: OR 0.33 [95% CI 0.16-0.66]) as was mean hospital cost (four studies: MD -7.38 thousand US Dollars or Euros [95% CI -14.14 to -0.62]).
CONCLUSIONS: Initiation of appropriate versus inappropriate antibiotics can reduce mortality, reduce treatment failure and decrease LOS, highlighting the importance of broad‑spectrum empiric therapy and rapid diagnostics for early identification of the causative pathogen.
STUDY REGISTRATION: PROSPERO: CRD42018104669.

PMID: 33045353 [PubMed - as supplied by publisher]

The current state of immunization against Gram-negative bacteria in children: a review of the literature.

Curr Opin Infect Dis. 2020 Oct 09;:

Authors: Broad J, Le Doare K, Heath PT, Hallchurch P, Whelan I, Boyd H, Carruthers E, Sharland M, Ladhani S

Abstract
PURPOSE OF REVIEW: Gram-negative bacteria (GNB) are a major cause of infection worldwide and multidrug resistance in infants and children. The major pathogens include Klebsiella pneumoniae, Escherichia coli, Enterobacter spp., Pseudomonas aeruginosa and Acinetobacter baumannii. With new antibiotic options limited, immunization is likely to play a critical role in prevention. This review discusses their epidemiology, the current state of vaccine research and potential immunization strategies to protect children. A comprehensive review of the literature, conference abstracts along with web searches was performed to identify current and investigational vaccines against the major GNB in children.
RECENT FINDINGS: Phase I--III vaccine trials have been undertaken for the major Gram-negative bacteria but not in infants or children. E. coli is a common infection in immune-competent children, including neonatal sepsis. Several vaccines are in late-phase clinical trials, with some already licensed for recurrent urinary tract infections in women. Klebsiella spp. causes community-acquired and hospital-acquired infections, including sepsis in neonates and immunocompromised children although no vaccine trials have extended beyond early phase 2 trials. P. aeruginosa is a common pathogen in patients with cystic fibrosis. Phase 1--3 vaccine and monoclonal antibody trials are in progress, although candidates provide limited coverage against pathogenic strains. Enterobacter spp. and A. baumannii largely cause hospital-acquired infections with experimental vaccines limited to phase 1 research.
SUMMARY: The current immunization pipelines for the most prevalent GNB are years away from licensure. Similar to incentives for new antibiotics, global efforts are warranted to expedite the development of effective vaccines.

PMID: 33044242 [PubMed - as supplied by publisher]

Current transition management of adolescents and young adults with allergy and asthma: a European survey.

Clin Transl Allergy. 2020;10:40

Authors: Khaleva E, Vazquez-Ortiz M, Comberiati P, DunnGalvin A, Pite H, Blumchen K, Garriga-Baraut T, Hox V, Santos AF, Gore C, Knibb RC, Alviani C, Mortz CG, Angier E, Duca B, Jensen B, Sanchez-Garcia S, Gowland MH, Timmermans F, Pfaar O, Roberts G

Abstract
Background: Transition from parent-delivered to self-management is a vulnerable time for adolescents and young adults (AYA) with allergy and asthma. There is currently no European guideline available for healthcare professionals (HCPs) on transition of these patients and local/national protocols are also mostly lacking.
Methods: European HCPs working with AYA with allergy and asthma were invited to complete an online survey assessing challenges of working with these patients, current transition practices and access to specific healthcare resources.
Results: A total of 1179 responses from 41 European countries were collected. Most HCPs (86%) reported a lack of a transition guideline and a lack of a transition process (20% paediatric HCPs, 50% of adult HCPs, 56% HCP seeing all ages). Nearly half (48%) acknowledged a lack of an established feedback system between paediatric and adult medical services. Many respondents never routinely asked about mental health issues such as self-harm or depression and are not confident in asking about self-harm (66.6%), sexuality (64%) and depression (43.6%). The majority of HCPs (76%) had not received specific training in the care of AYA although 87% agreed that transition was important for AYA with allergy and asthma.
Conclusion: Although there was agreement that transition is important for AYA with allergy and asthma, there are crucial limitations and variations in the current provision of transition services across Europe. Standardisation of AYA management and specific training are required. This should improve management and continuity of care during adolescence and into adulthood to achieve the best healthcare outcomes.

PMID: 33042515 [PubMed]

A Novel Mouse Model of Aminoglycoside-Induced Hyperacusis and Tinnitus.

Front Neurosci. 2020;14:561185

Authors: Longenecker RJ, Gu R, Homan J, Kil J

Abstract
Aminoglycosides (AG) such as amikacin are commonly used in cystic fibrosis patients with opportunistic pulmonary infections including multi-drug resistant mycobacterium tuberculous and non-tuberculous mycobacterium. Unfortunately, this class of drugs is known to cause peripheral damage to the cochlea leading to hearing loss that can fluctuate and become permanent over time or multiple exposures. However, whether amikacin can lead to central auditory dysfunction like hyperacusis (increased sensitivity to sound) or tinnitus (perception of sound in the absence of acoustic stimulation) is not well-described in the literature. Thus, an animal model needs to be developed that documents these side effects in order to develop therapeutic solutions to reduce AG-induced auditory dysfunction. Here we present pioneer work in mice which demonstrates that amikacin can lead to fluctuating behavioral evidence of hyperacusis and tinnitus as assessed by the acoustic startle reflex. Additionally, electrophysiological assessments of hearing via auditory brainstem response demonstrate increased central activity in the auditory brainstem. These data together suggest that peripheral AG-induced dysfunction can lead to central hyperactivity and possible behavioral manifestations of hyperacusis and tinnitus. Importantly, we demonstrate that ebselen, a novel investigational drug that acts as both an antioxidant and anti-inflammatory, can mitigate AG-induced hyperacusis.

PMID: 33041759 [PubMed]

Real World Experience of Pseudomonas aeruginosa Eradication at an Urban Pediatric Cystic Fibrosis Center.

J Pediatr Pharmacol Ther. 2020;25(7):623-628

Authors: Jablonski L, Lee CKK, Rosenstein BJ, Mogayzel PJ, Paranjape S, Pan A

Abstract
OBJECTIVES: Clinical practice guidelines for eradication of Pseudomonas aeruginosa (PA) in patients with cystic fibrosis (CF) have been established, but current studies have not assessed how these guidelines translate into clinical practice. This study aimed to characterize the real-world eradication strategies, eradication rates, and microbiologic outcomes of patients with first acquisition of PA at an urban pediatric CF center.
METHODS: The Cystic Fibrosis Foundation Patient Registry was used to identify patients with CF who received care between January 2014 and September 2018 and had PA isolated from an airway culture. Patients were included if they had a first positive PA culture or the first positive culture in 2 years. Data regarding patient demographics, timing and results of airway cultures, and treatment regimens were collected.
RESULTS: Over a 3.75-year period, 75 patients had an initial positive culture for PA. Of those patients, 74 (98.7%) received eradication treatment. Tobramycin inhalation solution (TIS) monotherapy was the most common regimen prescribed (52.7%) followed by TIS plus an oral fluoroquinolone (28.4%) (TIS + FQ). Of those treated, 62 (83.8%) patients had eradication of PA at first follow-up culture (median, 58 days; IQR, 49-77 days). Eradication rates (84.6% vs 76.2%, p = 0.421) and times to recurrence (6.37 months vs 5.1 months, p = 0.726) were comparable between TIS and TIS + FQ cohorts.
CONCLUSIONS: The eradication rate for PA in clinical practice is similar to that published in the literature. Consistent with published guidelines, these microbiologic outcomes do not support the addition of an oral FQ to TIS for initial PA eradication.

PMID: 33041717 [PubMed]

A Much-Anticipated Leap Forward in β-Lactam Drug Allergy: Phenotyping Reactions to Piperacillin/Tazobactam.

J Allergy Clin Immunol Pract. 2020 Oct;8(9):3178-3179

Authors: Madrigal-Burgaleta R, Fernandes B, Watson D, Shafi N, Ali FR

PMID: 33039017 [PubMed - as supplied by publisher]

Long-term docosahexaenoic acid (DHA) supplementation in cystic fibrosis patients: a randomized, multi-center, double-blind, placebo-controlled trial.

Prostaglandins Leukot Essent Fatty Acids. 2020 Oct 01;162:102186

Authors: López-Neyra A, Suárez L, Muñoz M, de Blas A, Ruiz de Valbuena M, Garriga M, Calvo J, Ribes C, Girón Moreno R, Máiz L, González D, Bousoño C, Manzanares J, Pastor Ó, Martínez-Botas J, Del Campo R, Cantón R, Roy G, Menacho M, Arroyo D, Zamora J, Soriano JB, Lamas A

Abstract
BACKGROUND: Cystic fibrosis (CF) patients have an alteration in fatty acid (FA) metabolism, associated with increased omega-6 and low omega-3 FA. Previous studies on supplementation with omega-3 FA in CF had contradictory results, and to date there is no evidence to recommend routine use of omega-3 supplements in CF patients. We hypothesized that long-term supplementation with docosahexaenoic acid (DHA) will have beneficial effects in these patients, by reducing pulmonary, systemic and intestinal inflammation.
METHODS: This was a randomized, double-blind, parallel, placebo-controlled trial. CF patients (age >2 months) were randomized to receive a seaweed DHA oil solution (50 mg/Kg/day) or matching placebo for 48 weeks. Primary outcomes were pulmonary (interleukin [IL]-8), systemic (IL-8) and intestinal (calprotectin) inflammatory biomarkers. Secondary outcomes included other pulmonary (IL-1β, IL-6, neutrophil elastase, lactate and calprotectin) and systemic (serum-IL-1β, IL-6) inflammatory biomarkers, as well as clinical outcomes (FEV1, pulmonary exacerbations, antibiotic use, nutritional status and quality of life).
RESULTS: Ninety six CF patients, 44 female, age 14.6±11.9 years (48 DHA and 48 placebo) were included. At trial completion, there were no differences in all primary outcomes [serum-IL-8 (p=0.909), respiratory-IL-8 (p=0.384) or fecal calprotectin (p=0.948)], all secondary inflammatory biomarkers, or in any of the clinical outcomes evaluated. There were few adverse events, with similar incidence in both study groups.
CONCLUSION: In this study, long-term DHA supplementation in CF patients was safe, but did not offer any benefit on inflammatory biomarkers, or in clinical outcomes compared with placebo. (NCT01783613).

PMID: 33038833 [PubMed - as supplied by publisher]

Glued in lipids: Lipointoxication in cystic fibrosis.

EBioMedicine. 2020 Oct 07;61:103038

Authors: Vandebrouck C, Ferreira T

Abstract
Cystic Fibrosis (CF) is an autosomal recessive disease caused by mutations in the CF transmembrane regulator (CFTR) gene, which encodes a chloride channel located at the apical surface of epithelial cells. Unsaturated Fatty Acid (UFA) deficiency has been a persistent observation in tissues from patients with CF. However, the impacts of such deficiencies on the etiology of the disease have been the object of intense debates. The aim of the present review is first to highlight the general consensus on fatty acid dysregulations that emerges from, sometimes apparently contradictory, studies. In a second step, a unifying mechanism for the potential impacts of these fatty acid dysregulations in CF cells, based on alterations of membrane biophysical properties (known as lipointoxication), is proposed. Finally, the contribution of lipointoxication to the progression of the CF disease and how it could affect the efficacy of current treatments is also discussed.

PMID: 33038767 [PubMed - as supplied by publisher]

Effect of apical chloride concentration on the measurement of responses to CFTR modulation in airway epithelia cultured from nasal brushings.

Physiol Rep. 2020 Oct;8(19):e14603

Authors: Bratcher PE, Yadav S, Shaughnessy CA, Thornell IM, Zeitlin PL

Abstract
INTRODUCTION: One method for assessing the in vitro response to CFTR-modulating compounds is by analysis of epithelial monolayers in an Ussing chamber, where the apical and basolateral surfaces are isolated and the potential difference, short-circuit current, and transepithelial resistance can be monitored. The effect of a chloride ion gradient across airway epithelia on transepithelial chloride transport and the magnitude of CFTR modulator efficacy were examined.
METHODS: CFTR-mediated changes in the potential difference and transepithelial currents of primary human nasal epithelial cell cultures were quantified in Ussing chambers with either symmetrical solutions or reduced chloride solutions in the apical chamber. CFTR activity in homozygous F508del CFTR epithelia was rescued by treatment with VX-661, C4/C18, 4-phenylbutyrate (4-PBA) for 24 hr at 37°C or by incubation at 29°C for 48 hr.
RESULTS: Imposing a chloride gradient increased CFTR-mediated and CaCC-mediated ion transport. Treatment of F508del CFTR homozygous cells with CFTR modulating compounds increased CFTR activity, which was significantly more evident in the presence of a chloride gradient. This observation was recapitulated with temperature-mediated F508del CFTR correction.
CONCLUSIONS: Imposing a chloride gradient during Ussing chamber measurements resulted in increased CFTR-mediated ion transport in expanded non-CF and F508del CFTR homozygous epithelia. In F508del CFTR homozygous epithelia, the magnitude of response to CFTR modulating compounds or low temperature was greater when assayed with a chloride gradient compared to symmetrical chloride, resulting in an apparent increase in measured efficacy. Future work may direct which methodologies utilized to quantify CFTR modulator response in vitro are most appropriate for the estimation of in vivo efficacy.

PMID: 33038073 [PubMed - as supplied by publisher]

COVID-19 multidisciplinary high dependency unit: the Milan model.

Respir Res. 2020 Oct 09;21(1):260

Authors: Aliberti S, Amati F, Pappalettera M, Di Pasquale M, D'Adda A, Mantero M, Gramegna A, Simonetta E, Oneta AM, Privitera E, Gori A, Bozzi G, Peyvandi F, Minoia F, Filocamo G, Abbruzzese C, Vicenzi M, Tagliabue P, Alongi S, Blasi F

Abstract
COVID-19 is a complex and heterogeneous disease. The pathogenesis and the complications of the disease are not fully elucidated, and increasing evidence shows that SARS-CoV-2 causes a systemic inflammatory disease rather than a pulmonary disease. The management of hospitalized patients in COVID-19 dedicated units is advisable for segregation purpose as well as for infection control. In this article we present the standard operating procedures of our COVID-19 high dependency unit of the Policlinico Hospital, in Milan. Our high dependency unit is based on a multidisciplinary approach. We think that the multidisciplinary involvement of several figures can better identify treatable traits of COVID-19 disease, early identify patients who can quickly deteriorate, particularly patients with multiple comorbidities, and better manage complications related to off-label treatments. Although no generalizable to other hospitals and different healthcare settings, we think that our experience and our point of view can be helpful for countries and hospitals that are now starting to face the COVID-19 outbreak.

PMID: 33036610 [PubMed - as supplied by publisher]