Decreased Electrogenic Anionic Secretory Response in the Porcine Colon Following in vivo Challenge with Brachyspira spp. Supports an Altered Mucin Environment.

Am J Physiol Gastrointest Liver Physiol. 2019 Jan 10;:

Authors: Enns CB, Harding JCS, Loewen ME

Abstract
Brachyspiraspp. causediarrheal disease in multiple animal species by colonization of the colon, resulting in colitis, mucus induction and disrupted ion transport. Unique to spirochete pathogenesisis the immense production of mucus resulting in a niche mucin environment likely favoring spirochete colonization. Mucin rheological properties are heavily influencedby anionic secretion and loss of secretory function has been implicated in diseases such as cystic fibrosis. Here, the effects on the agonist-inducedelectrogenicanionic secretory response by infectious colonic spirochete bacteria Brachyspirahyodysenteriaeand Brachyspira hampsoniiwere assessedin the proximal, apex and distal sections of colonin Ussing chambers. Activation of secretion via isoproterenol, carbachol, andforskolin / IBMX demonstrated a significantly decreased change in short-circuit current ( Isc) in Brachyspira-infected pigs in all sections. Tissue resistances did not account for this difference, rather, it was attributed to a decrease in anionic secretion as indicated by a decrease in bumetanideinhibitable Isc. RT-qPCR analyses and western blot determined the major anionic channels of the epithelium were down-regulated in diarrheic pigs paired with altered mucin gene expression. The investigated cytokines were not responsible for the down-regulation of anion channel gene transcripts. Although IL-1α was upregulatedin all segments,it did not alter CFTR mRNA expression in Caco-2 monolayers. However, a whole cell Brachyspirahampsoniilysate significantly reduced CFTR mRNA expression in Caco-2 monolayers. Together, these findings indicate that these two Brachyspira spp. may directlycause a decreased anionic secretory response in the porcine colon supporting an altered mucin environment likely favoring spirochete colonization.

PMID: 30629469 [PubMed - as supplied by publisher]

[New treatments in respiratory medicine in 2018 : asthma, cystic fibrosis and nocturnal positive pressure (CPAP or NIV)].

Rev Med Suisse. 2019 Jan 09;15(N° 632-633):96-100

Authors: Plojoux J, Charbonnier F, Janssens JP

Abstract
In this review of novel therapies in pulmonary disorders in 2018, we cover 3 different entities. In GINA stage 1 and 2 asthma, new strategies allow a more individualized treatment. In more severe asthma, there is an increasing interest in biotherapies, with dupilumab, an anti-IL-4 receptor, completing the already available panel which includes anti-IgE, anti-IL-5 and anti-IL-5 receptor antibodies. In cystic fibrosis, a better understanding of the pathogenesis and the genetics of the disease is leading to new treatments acting directly on the function of the CFTR (Cystic fibrosis transmembrane conductance regulator), which, when used in combination, show very promising results. Finally, nocturnal positive pressure treatment (either CPAP or non invasive ventilation : NIV) is used more and more frequently. Recent studies have clarified therapeutic algorithms for the most frequent indications for NIV : COPD and obesity-hypoventilation.

PMID: 30629379 [PubMed - in process]

Burkholderia cenocepacia utilizes a type VI secretion system for bacterial competition.

Microbiologyopen. 2019 Jan 09;:e774

Authors: Spiewak HL, Shastri S, Zhang L, Schwager S, Eberl L, Vergunst AC, Thomas MS

Abstract
Burkholderia cenocepacia is an opportunistic bacterial pathogen that poses a significant threat to individuals with cystic fibrosis by provoking a strong inflammatory response within the lung. It possesses a type VI secretion system (T6SS), a secretory apparatus that can perforate the cellular membrane of other bacterial species and/or eukaryotic targets, to deliver an arsenal of effector proteins. The B. cenocepacia T6SS (T6SS-1) has been shown to be implicated in virulence in rats and contributes toward actin rearrangements and inflammasome activation in B. cenocepacia-infected macrophages. Here, we present bioinformatics evidence to suggest that T6SS-1 is the archetype T6SS in the Burkholderia genus. We show that B. cenocepacia T6SS-1 is active under normal laboratory growth conditions and displays antibacterial activity against other Gram-negative bacterial species. Moreover, B. cenocepacia T6SS-1 is not required for virulence in three eukaryotic infection models. Bioinformatics analysis identified several candidate T6SS-dependent effectors that may play a role in the antibacterial activity of B. cenocepacia T6SS-1. We conclude that B. cenocepacia T6SS-1 plays an important role in bacterial competition for this organism, and probably in all Burkholderia species that possess this system, thereby broadening the range of species that utilize the T6SS for this purpose.

PMID: 30628184 [PubMed - as supplied by publisher]

[Patients with cystic fibrosis become adults : Treatment hopes and disappointments].

Internist (Berl). 2019 Jan 09;:

Authors: Eschenhagen P, Schwarz C

Abstract
Mucoviscidosis or cystic fibrosis (CF) is one of the most frequent monogenetic diseases in middle Europe. It is inherited in an autosomal recessive manner. A defect in the cystic fibrosis transmembrane conductance regulator (CFTR) channel reduces chloride ion transport to the cell membrane, which leads to malfunctions in all exocrine glands. This results in a progressive multiorgan disease, which leads to chronic inflammation and infections of the lungs. The progressive destruction of lung tissue with respiratory insufficiency is the most common cause of death in CF. Progress in symptomatic treatment over the past decades has led to a dramatic improvement in life expectation and quality of life for those affected, so that nowadays in nearly all industrial countries the majority of patients are adults. In 2012 the era of causal therapy of the CFTR protein defects was opened with the approval of ivacaftor. Long-term data now confirm the benefits. There is reason to hope that the success story of CF treatment will be continued, particularly by further CFTR modulators with innovative modes of action and improved efficacy; however, so far these are not available for all mutation classes, so that not all patients can reap the benefits. Therefore, the further development of symptomatic treatment becomes of great importance due to the complications that have already occurred before the implementation of the CFTR modulators. The implementation of modulators in early childhood can attenuate or prevent early irreversible complications. Therefore, in this article special emphasis is placed on new developments in symptomatic treatment and on new treatment options.

PMID: 30627755 [PubMed - as supplied by publisher]

Curcumin down-regulates toll-like receptor-2 gene expression and function in human cystic fibrosis bronchial epithelial cells.

Biol Pharm Bull. 2019 Jan 10;:

Authors: Chaudhary N, Ueno-Shuto K, Ono T, Ohira Y, Watanabe K, Nasu A, Fujikawa H, Nakashima R, Takahashi N, Suico MA, Kai H, Shuto T

Abstract
Cystic fibrosis (CF), the most common lethal inherited disorder caused by mutation in the gene encoding the CF transmembrane regulator (CFTR), is characterized by chronic inflammation that ultimately leads to death from respiratory failure. In CF patients, up-regulation of toll-like receptor-2 (TLR2), a pattern recognition receptor that senses CF-pathogenic bacteria Staphylococcus aureus peptidoglycan (PGN), in airway epithelial cells is observed, and enhanced proinflammatory responses towards PGN may result in detrimental effects in CF patients. Here, we showed that curcumin, a well known anti-inflammatory agent derived from the curry spice turmeric, inhibits TLR2 expression in CF bronchial epithelial cell line, CFBE41o- cells. Strong suppression of TLR2 gene and protein expression was observed at more than 40 μM of curcumin treatment in CFBE41o- cells. Consistent with decreased expression of TLR2, PGN-dependent IL-8 gene up-regulation was markedly reduced by 40 μM of curcumin treatment. Strong reductions of TLR2 gene expression and function were also observed in primary human CF bronchial epithelial cells, but not in human non-CF primary cells. Interestingly, curcumin treatment decreased nuclear expression of transcription factor specificity protein 1 (SP1), a factor that is critical for increased basal TLR2 expression in CF cell line and primary cells. Finally, curcumin-dependent SP1 reduction was diminished by anti-oxidant N-acetylcystein (NAC) and proteasomal inhibitor MG-132, suggesting the crucial roles of oxidative and proteasomal degradation pathways. Taken together, our study shows that curcumin down-regulates TLR2 gene expression and function in CF bronchial epithelial cells possibly by accelerating SP1 degradation via an oxidative process.

PMID: 30626802 [PubMed - as supplied by publisher]

Allocating healthcare resources-seriously ill people should have priority.

BMJ. 2018 04 27;361:k1817

Authors: Segall M

PMID: 29703725 [PubMed - indexed for MEDLINE]

Validation of multiple breath washout devices.

J Cyst Fibros. 2017 11;16(6):e22-e23

Authors: Raaijmakers L, Jensen R, Stanojevic S, Ratjen F

PMID: 28185888 [PubMed - indexed for MEDLINE]

Can secretory immunoglobulin A in saliva predict a change in lung infection status in patients with cystic fibrosis? A prospective pilot study.

Health Sci Rep. 2018 Aug;1(8):e52

Authors: Alanin MC, Pressler T, Aanaes K, Ekstrøm CT, Skov M, Johansen HK, Nielsen KG, von Buchwald C, Høiby N

Abstract
Background: Chronic lung infection with Pseudomonas aeruginosa is the main cause of mortality in patients with cystic fibrosis (CF). Sinus colonization with P. aeruginosa often precedes intermittent lung colonization, and intermittent colonization precedes chronic infection.When P. aeruginosa colonizes the sinuses, elevated immunoglobulin A (IgA) levels specific against P. aeruginosa can be detected in saliva. Therefore, we hypothesized that increasing levels of IgA in saliva can be detected before P. aeruginosa lung colonization.
Methods: Forty-nine CF patients free from lung colonization with P. aeruginosa or other Gram-negative bacteria (GNB) were included in this prospective study. Saliva and serum samples were collected and examined for IgA antibodies against P. aeruginosa with at least 6-month intervals between sequential samples.
Results: A total of 110 measurements of IgA in saliva were included. During a median of 8.5-month follow-up, 25 patients changed their lung infection status. We were able to construct a statistical model that for a given value of IgA in saliva, could predict the probability of a change in lung infection status within the next 8.5 months (median): p = 1 / (1 + exp(-(-0.9582 + 1.6518*IgA)). The model includes a prediction band where 95% of new measurements are predicted to fall within. The model, however, failed to reach statistical significance (P = 0.056 1-tailed), probably because of lack of power.
Conclusion: The saliva IgA model may predict a worsening in lung infection status presumably acting as a surrogate marker of P. aeruginosa bacterial sinusitis. The model may identify patients at risk of subsequent lung colonization and, thus, be a helpful clinical tool, but it should be tested in studies with larger sample sizes to evaluate its utility.

PMID: 30623088 [PubMed]

Sweat chloride quantification using capillary electrophoresis.

Pract Lab Med. 2019 Jan;13:e00114

Authors: Dubot P, Liang J, Dubs J, Missiak Y, Sarazin C, Couderc F, Caussé E

Abstract
Background: Cystic fibrosis (CF) is the less rare and severe genetic disease among the European population. Biochemical diagnosis of CF is based on the demonstration of increased chloride concentration in sweat samples, obtained during the sweat test (ST). WynSep developed a capillary electrophoresis with contactless conductivity detection (CE-C4D) able to measure sweat chloride with a low sample volume. We evaluated the clinical feasibility of this device in a cohort of patients suspected of CF, in comparison with the common coulometric method (ChloroChek chloridometer).
Methods: We determined sweat chloride concentration of 65 samples from patients referred to our institution to undergo a sweat test. Each sample was submitted to coulometric method first, then WynSep-CE, with or without internal standard (IS) subject to sufficient volume sample.
Results: A total of 53 samples were analysed by both coulometric and WynSep-CE (using IS) methods. The method validation showed comparable analytical performances for both methods; no false positive or false negative was recorded. The two methods showed a high correlation (r = 0.993, p < 0.001) and a close agreement was demonstrated by two different statistical tests (Bland-Altman and Passing-Bablok).
Conclusions: WynSep-CE is an accurate, fast, easy-to-use and an appropriate method for CF diagnosis.

PMID: 30623007 [PubMed]

Current therapies and novel approaches for biliary diseases.

World J Gastrointest Pathophysiol. 2019 Jan 05;10(1):1-10

Authors: Rajapaksha IG, Angus PW, Herath CB

Abstract
Chronic liver diseases that inevitably lead to hepatic fibrosis, cirrhosis and/or hepatocellular carcinoma have become a major cause of illness and death worldwide. Among them, cholangiopathies or cholestatic liver diseases comprise a large group of conditions in which injury is primarily focused on the biliary system. These include congenital diseases (such as biliary atresia and cystic fibrosis), acquired diseases (such as primary sclerosing cholangitis and primary biliary cirrhosis), and those that arise from secondary damage to the biliary tree from obstruction, cholangitis or ischaemia. These conditions are associated with a specific pattern of chronic liver injury centered on damaged bile ducts that drive the development of peribiliary fibrosis and, ultimately, biliary cirrhosis and liver failure. For most, there is no established medical therapy and, hence, these diseases remain one of the most important indications for liver transplantation. As a result, there is a major need to develop new therapies that can prevent the development of chronic biliary injury and fibrosis. This mini-review briefly discusses the pathophysiology of liver fibrosis and its progression to cirrhosis. We make a special emphasis on biliary fibrosis and current therapeutic options, such as angiotensin converting enzyme-2 (known as ACE2) over-expression in the diseased liver as a novel potential therapy to treat this condition.

PMID: 30622832 [PubMed]

The ionophore oxyclozanide enhances tobramycin killing of Pseudomonas aeruginosa biofilms by permeabilizing cells and depolarizing the membrane potential.

J Antimicrob Chemother. 2019 Jan 08;:

Authors: Maiden MM, Zachos MP, Waters CM

Abstract
Objectives: To assess the ability of oxyclozanide to enhance tobramycin killing of Pseudomonas aeruginosa biofilms and elucidate its mechanism of action.
Methods: Twenty-four hour biofilms formed by the P. aeruginosa strain PAO1 and cystic fibrosis (CF) isolates were tested for susceptibility to oxyclozanide and tobramycin killing using BacTiter-Glo™ and cfu. Biofilm dispersal was measured using crystal violet staining. Membrane potential and permeabilization were quantified using DiOC2(3) and TO-PRO-3, respectively.
Results: Here we show that the ionophore anthelmintic oxyclozanide, combined with tobramycin, significantly increased killing of P. aeruginosa biofilms over each treatment alone. This combination also significantly accelerated the killing of cells within biofilms and stationary phase cultures and it was effective against 4/6 CF clinical isolates tested, including a tobramycin-resistant strain. Oxyclozanide enhanced the ability of additional aminoglycosides and tetracycline to kill P. aeruginosa biofilms. Finally, oxyclozanide permeabilized cells within the biofilm, reduced the membrane potential and increased tobramycin accumulation within cells of mature P. aeruginosa biofilms.
Conclusions: Oxyclozanide enhances aminoglycoside and tetracycline activity against P. aeruginosa biofilms by reducing membrane potential, permeabilizing cells and enhancing tobramycin accumulation within biofilms. We propose that oxyclozanide counteracts the adaptive resistance response of P. aeruginosa to aminoglycosides, increasing both their maximum activity and rate of killing. As oxyclozanide is widely used in veterinary medicine for the treatment of parasitic worm infections, this combination could offer a new approach for the treatment of biofilm-based P. aeruginosa infections, repurposing oxyclozanide as an anti-biofilm agent.

PMID: 30624737 [PubMed - as supplied by publisher]

Adenylyl cyclase 6 expression is essential for cholera toxin-induced diarrhea.

J Infect Dis. 2019 Jan 08;:

Authors: Fenton RA, Murali SK, Kaji I, Akiba Y, Kaunitz JD, Kristensen TB, Poulsen SB, Dominguez Rieg JA, Rieg T

Abstract
Background: Cholera toxin (CT)-induced diarrhea is mediated by cAMP-mediated active Cl - secretion via the cystic fibrosis transmembrane conductance regulator (CFTR). Although the constitutive activation of adenylyl cyclase (AC) in response to CT is due to ADP-ribosylation of Gsα activating CFTR with consequent secretory diarrhea, the AC isoform(s) involved remain unknown.
Methods: We generated intestine epithelial cell-specific adenylyl cyclase 6 (AC6) knockout mice to study its role in cholera toxin-induced diarrhea.
Results: AC6 mRNA levels were the highest of all 9 membrane-bound AC isoforms in mouse intestinal epithelial cells. Intestinal epithelial-specific AC6 knockout mice (AC6loxloxVillinCre) had undetectable AC6 levels in small intestinal and colonic epithelial cells. No significant differences in fluid and food intake, plasma electrolytes, intestinal/colon anatomy and morphology, as well as fecal water content were observed between genotypes. Nevertheless, CT-induced fluid accumulation in vivo was completely absent in AC6loxloxVillinCre mice, associated with a lack of forskolin- and CT-induced changes in short-circuit current (Isc) of the intestinal mucosa, impaired cAMP generation in acutely isolated small intestinal epithelial cells and significantly impaired apical CFTR levels in response to forskolin.
Conclusion: AC6 is a novel target for the treatment of CT-induced diarrhea.

PMID: 30624615 [PubMed - as supplied by publisher]

Newborn Screening Follow-up.

N C Med J. 2019 Jan-Feb;80(1):37-41

Authors: Percenti L, Vickery G

Abstract
Newborn screening identifies infants at increased risk for 50 metabolic conditions that are treatable with early detection. Follow-up links the laboratory result with physician action. Timeliness is essential to ensure affected babies receive treatment and enjoy a healthy and productive life. The follow-up process comprises many steps, several disciplines, and various challenges.

PMID: 30622203 [PubMed - in process]

Sleep-Related Rhythmic Movement Disorder in Triplets: Evidence for Genetic Predisposition?

J Clin Sleep Med. 2019 Jan 03;:

Authors: Hayward-Koennecke HK, Werth E, Valko PO, Baumann CR, Poryazova R

Abstract
ABSTRACT: Sleep-related rhythmic movement disorder (RMD) is common in very young children but rarely persists beyond childhood. Despite its high frequency, the underlying pathophysiology remains unclear. Familial occurrence is rare. Here we present monozygotic female triplets, all of them being affected by body rolling in terms of RMD. Furthermore, they all present with an additional genetic disease, cystic fibrosis, with the same documented mutation of the cystic fibrosis transmembrane conductance regulator gene (F508del-CFTR). Because all three monozygotic siblings are concordant for RMD, genetic factors may contribute to the time course of the disorder.

PMID: 30621834 [PubMed - as supplied by publisher]

Mortality Due to Cystic Fibrosis over a 36-Year Period in Spain: Time Trends and Geographic Variations.

Int J Environ Res Public Health. 2019 Jan 04;16(1):

Authors: Villaverde-Hueso A, Sánchez-Díaz G, Molina-Cabrero FJ, Gallego E, Posada de la Paz M, Alonso-Ferreira V

Abstract
The aim of this study is to analyze population-based mortality attributed to cystic fibrosis (CF) over 36 years in Spain. CF deaths were obtained from the National Statistics Institute, using codes 277.0 from the International Classification of Diseases (ICD) ninth revision (ICD9-CM) and E84 from the tenth revision (ICD10) to determine the underlying cause of death. We calculated age-specific and age-adjusted mortality rates, and time trends were assessed using joinpoint regression. The geographic analysis by district was performed by standardized mortality ratios (SMRs) and smoothed-SMRs. A total of 1002 deaths due to CF were identified (50.5% women). Age-adjusted mortality rates fell by -0.95% per year between 1981 and 2016. The average age of death from CF increased due to the annual fall in the mortality of under-25s (-3.77% males, -2.37% females) and an increase in over-75s (3.49%). We identified districts with higher than expected death risks in the south (Andalusia), the Mediterranean coast (Murcia, Valencia, Catalonia), the West (Extremadura), and the Canary Islands. In conclusion, in this study we monitored the population-based mortality attributed to CF over a long period and found geographic differences in the risk of dying from this disease. These findings complement the information provided in other studies and registries and will be useful for health planning.

PMID: 30621191 [PubMed - in process]

A fungal metabolite zearalenone as a CFTR inhibitor and potential therapy of secretory diarrheas.

Biochem Pharmacol. 2018 04;150:293-304

Authors: Muangnil P, Satitsri S, Tadpetch K, Saparpakorn P, Chatsudthipong V, Hannongbua S, Rukachaisirikul V, Muanprasat C

Abstract
Overstimulation of CFTR-mediated Cl- secretion plays an important role in the pathogenesis of secretory diarrheas, which remain an important global health problem. This study aimed to identify inhibitors of CFTR-mediated Cl- secretion from a library of fungus-derived compounds and to evaluate their pharmacological properties and anti-diarrheal utility. We identified zearalenone, 7'-dehydrozearalenone and 8'-hydroxyzearalenone isolated from the seagrass-derived fungus Fusarium sp. PSU-ES123 as inhibitors of CFTR-mediated Cl- secretion in human intestinal epithelial (T84) cells. Being the most potent fungal metabolite capable of inhibiting CFTR-mediated Cl- secretion, zearalenone reversibly inhibited CFTR Cl- channel activity in T84 cells with an IC50 of ∼0.5 μM. Functional and biochemical analyses and molecular docking studies indicate that zearalenone binds to the β-estradiol binding sites in the ATP-binding pockets on NBD1 and NBD2 of CFTR. Mechanisms of CFTR inhibition by zearalenone do not involve activation of phosphodiesterases, protein phosphatases, multidrug-resistance protein 4 and AMP-activated protein kinases. Importantly, zearalenone significantly inhibited cholera toxin (CT)-induced Cl- secretion in T84 cells and blocked CT-induced intestinal fluid secretion in mice. Collectively, our study indicates that zearalenone represents the first class of fungus-derived CFTR inhibitors. Further development of this class of compounds may provide an effective treatment of secretory diarrheas.

PMID: 29475061 [PubMed - indexed for MEDLINE]

Pseudomonas aeruginosa eradication: Finally moving the needle?

J Cyst Fibros. 2017 05;16(3):309-310

Authors: Cogen J, Rosenfeld M

PMID: 28416146 [PubMed - indexed for MEDLINE]

The patient voice in research - Supporting actor or starring role?

J Cyst Fibros. 2017 05;16(3):313-314

Authors: Rowbotham NJ, Smyth AR

PMID: 28285933 [PubMed - indexed for MEDLINE]

Counterpoint: Confronting the challenge of non-adherence: Building patient-provider relationships - a patient's perspective.

J Cyst Fibros. 2017 03;16(2):306-307

Authors: Kvam CM

PMID: 28238635 [PubMed - indexed for MEDLINE]

Counterpoint: Too little care or too little collaboration: Approaches to treatment refusal in CF.

J Cyst Fibros. 2017 03;16(2):304-305

Authors: Sawicki GS, Riekert KA

PMID: 28236582 [PubMed - indexed for MEDLINE]