Int J Environ Res Public Health. 2022 Apr 23;19(9):5155. doi: 10.3390/ijerph19095155.

ABSTRACT

This study sought to investigate the association of light physical activity (LPA), moderate-to-vigorous physical activity (MVPA), sedentary time (SED), and sleep with lung function in children and adults with CF. In total, 86 children (41 females; 13.6 ± 2.8 years; FEV1%predicted: 86 ± 1%) and 43 adults (21 females; 24.6 ± 4.7 years; FEV1%predicted: 63 ± 21%) with CF participated in this study. Wrist-worn accelerometery was used to assess PA, SED and sleep. Compositional linear regression models were conducted following normalisation via isometric log-ratio transformations. Sequential binary partitioning was applied to investigate the impact of reallocating 10 to 30 min between each behaviour on FEV1%predicted. A decline in FEV1%predicted was predicted with the reallocation of 30 min from MVPA to SED or LPA or sleep to any other behaviour in children (-3.04--0.005%) and adults (-3.58--0.005%). Conversely, improvements in FEV1%predicted were predicted when 30 min was reallocated to MVPA from LPA or SED in children (0.12-1.59%) and adults (0.77-2.10%), or when 30 min was reallocated to sleep from any other behaviour in both children (0.23-2.56%) and adults (1.08-3.58%). This study supports the importance of MVPA and sleep for maintaining and promoting lung function in people with CF.

PMID:35564550 | DOI:10.3390/ijerph19095155

Foods. 2022 Apr 19;11(9):1189. doi: 10.3390/foods11091189.

ABSTRACT

The concentration of thiocyanate (SCN-) in bodily fluids is a good indicator of potential and severe health issues such as nasal bleeding, goiters, vertigo, unconsciousness, several inflammatory diseases, and cystic fibrosis. Herein, a visual SCN- sensing method has been developed using the enzyme-like nature of positively charged gold quantum dots (Au QDs) mixed with 3,3',5,5'-tetramethylbenzidine (TMB) and hydrogen peroxide (H2O2). This research also reports a new method of synthesizing positively charged Au QDs directly from gold nanoparticles through a hydrothermal process. Microscopic imaging has showed that the Au QDs were 3-5 nm in size, and the emission wavelength was at 438 nm. Au QDs did not display any enzyme-like nature while mixed up with TMB and H2O2. However, the nanozymatic activity of Au QDs appeared when SCN- was included, leading to a very low detection limit (LOD) of 8 nM and 99-105% recovery in complex media. The steady-state kinetic reaction of Au QDs showed that Au QDs had a lower Michaelis-Menten constant (Km) toward H2O2 and TMB, which indicates that the Au QDs had a higher affinity for H2O2 and TMB than horseradish peroxidase (HRP). A mechanism study has revealed that the scavenging ability of hydroxyl (•OH) radicals by the SCN- group plays an important role in enhancing the sensitivity in this study. The proposed nanozymatic "Off-On" SCN- sensor was also successfully validated in commercial milk samples.

PMID:35563912 | DOI:10.3390/foods11091189

Cells. 2022 May 9;11(9):1587. doi: 10.3390/cells11091587.

ABSTRACT

Defective hydration of airway surface mucosa is associated with recurrent lung infection in cystic fibrosis (CF), a disease caused by CF transmembrane conductance regulator (CFTR) gene mutations. Whether the composition and/or presence of an airway surface liquid (ASL) is sufficient to prevent infection remains unclear. The susceptibility to infection of polarized wild type and CFTR knockdown (CFTR-KD) airway epithelial cells was determined in the presence or absence of a healthy ASL or physiological saline. CFTR-KD epithelia exhibited strong ASL volume reduction, enhanced susceptibility to infection, and reduced junctional integrity. Interestingly, the presence of an apical physiological saline alleviated disruption of the airway epithelial barrier by stimulating essential junctional protein expression. Thus, rehydrated CFTR-KD cells were protected from infection despite normally intense bacterial growth. This study indicates that an epithelial integrity gatekeeper is modulated by the presence of an apical liquid volume, irrespective of the liquid's composition and of expression of a functional CFTR.

PMID:35563895 | DOI:10.3390/cells11091587

Cells. 2022 Apr 27;11(9):1478. doi: 10.3390/cells11091478.

ABSTRACT

Bitter taste receptors (T2Rs) are G protein-coupled receptors (GPCRs) expressed in various cell types including ciliated airway epithelial cells and macrophages. T2Rs in these two innate immune cell types are activated by bitter products, including those secreted by Pseudomonas aeruginosa, leading to Ca2+-dependent activation of endothelial nitric oxide (NO) synthase (eNOS). NO enhances mucociliary clearance and has direct antibacterial effects in ciliated epithelial cells. NO also increases phagocytosis by macrophages. Using biochemistry and live-cell imaging, we explored the role of heat shock protein 90 (HSP90) in regulating T2R-dependent NO pathways in primary sinonasal epithelial cells, primary monocyte-derived macrophages, and a human bronchiolar cell line (H441). Immunofluorescence showed that H441 cells express eNOS and T2Rs and that the bitter agonist denatonium benzoate activates NO production in a Ca2+- and HSP90-dependent manner in cells grown either as submerged cultures or at the air-liquid interface. In primary sinonasal epithelial cells, we determined that HSP90 inhibition reduces T2R-stimulated NO production and ciliary beating, which likely limits pathogen clearance. In primary monocyte-derived macrophages, we found that HSP-90 is integral to T2R-stimulated NO production and phagocytosis of FITC-labeled Escherichia coli and pHrodo-Staphylococcus aureus. Our study demonstrates that HSP90 serves as an innate immune modulator by regulating NO production downstream of T2R signaling by augmenting eNOS activation without impairing upstream Ca2+ signaling. These findings suggest that HSP90 plays an important role in airway antibacterial innate immunity and may be an important target in airway diseases such as chronic rhinosinusitis, asthma, or cystic fibrosis.

PMID:35563784 | DOI:10.3390/cells11091478

Int J Mol Sci. 2022 May 6;23(9):5206. doi: 10.3390/ijms23095206.

ABSTRACT

Cystic fibrosis transmembrane conductance regulator (CFTR) is highly expressed on the ocular epithelium and plays a pivotal role in the fluid secretion driven by chloride transport. Dry eye disease is one of the most common diseases with limited therapeutic options. In this study, a high-throughput screening was performed to identify novel CFTR activators capable of inducing chloride secretion on the ocular surface. The screening of 50,000 small molecules revealed three novel CFTR activators. Among them, the most potent CFTR activator, Cact-3 (7-(3,4-dimethoxyphenyl)-N-(4-ethoxyphenyl)pyrazolo [1,5-α]pyrimidine-2-carboxamide), produced large and sustained Cl- currents in WT-CFTR-expressing FRT cells with no alterations of ANO1 and hERG channel activity. The application of Cact-3 strongly activated CFTR in the ocular epithelia of mice and it also significantly increased CFTR-mediated Cl- transport in a primary cultured human conjunctival epithelium. Cact-3 strongly stimulated tear secretion in normal mice. In addition, Cact-3 significantly reduced ocular surface damage and the expression of proinflammatory factors, including interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ in an experimental mouse model of dry eye disease. These results suggest that Cact-3, a novel CFTR activator, may be a potential development candidate for the treatment of dry eye disease.

PMID:35563597 | DOI:10.3390/ijms23095206

Int J Mol Sci. 2022 Apr 30;23(9):5029. doi: 10.3390/ijms23095029.

ABSTRACT

Drug repurposing is an attractive strategy for developing new antibacterial molecules. Herein, we evaluated the in vitro antibacterial, antibiofilm, and antivirulence activities of eight FDA-approved "non-antibiotic" drugs, comparatively to tobramycin, against selected Pseudomonas aeruginosa strains from cystic fibrosis patients. MIC and MBC values were measured by broth microdilution method. Time-kill kinetics was studied by the macro dilution method, and synergy studies were performed by checkerboard microdilution assay. The activity against preformed biofilms was measured by crystal violet and viable cell count assays. The effects on gene expression were studied by real-time quantitative PCR, while the cytotoxic potential was evaluated against IB3-1 bronchial CF cells. Ciclopirox, 5-fluorouracil, and actinomycin D showed the best activity against P. aeruginosa planktonic cells and therefore underwent further evaluation. Time-kill assays indicated actinomycin D and ciclopirox, contrarily to 5-fluorouracil and tobramycin, have the potential for bacterial eradication, although with strain-dependent efficacy. Ciclopirox was the most effective against the viability of the preformed biofilm. A similar activity was observed for other drugs, although they stimulate extracellular polymeric substance production. Ribavirin showed a specific antibiofilm effect, not dependent on bacterial killing. Exposure to drugs and tobramycin generally caused hyperexpression of the virulence traits tested, except for actinomycin D, which downregulated the expression of alkaline protease and alginate polymerization. Ciclopirox and actinomycin D revealed high cytotoxic potential. Ciclopirox and ribavirin might provide chemical scaffolds for anti-P. aeruginosa drugs. Further studies are warranted to decrease ciclopirox cytotoxicity and evaluate the in vivo protective effects.

PMID:35563420 | DOI:10.3390/ijms23095029

Int J Mol Sci. 2022 Apr 20;23(9):4560. doi: 10.3390/ijms23094560.

ABSTRACT

Burkholderia cenocepacia is an opportunistic pathogen that can lead to severe infections in patients suffering from cystic fibrosis (CF) and chronic granulomatous disease. Being an obligate aerobe, B. cenocepacia is unable to grow in the absence of oxygen. In this study, we show that the CF isolate B. cenocepacia H111 can survive in the absence of oxygen. Using a transposon sequencing (Tn-seq) approach, we identified 71 fitness determinants involved in anoxic survival, including a Crp-Fnr family transcriptional regulatory gene (anr2), genes coding for the sensor kinase RoxS and its response regulator RoxR, the sigma factor for flagella biosynthesis (FliA) and subunits of a cytochrome bd oxidase (CydA, CydB and the potentially novel subunit CydP). Individual knockouts of these fitness determinants significantly reduced anoxic survival, and inactivation of both anr copies is shown to be lethal under anoxic conditions. We also show that the two-component system RoxS/RoxR and FliA are important for virulence and swarming/swimming, respectively.

PMID:35562951 | DOI:10.3390/ijms23094560

FASEB J. 2022 May;36 Suppl 1. doi: 10.1096/fasebj.2022.36.S1.R4879.

ABSTRACT

BACKGROUND: The vitamin D receptor (VDR) is a nuclear receptor that is expressed in many tissues throughout the body, but is particularly abundant within the colon. VDR is classically known to be activated by the active form of vitamin D, calcitriol, but is appreciated to also serve as a receptor for luminal bile acids. VDR has previously been shown to have anti-inflammatory and barrier promoting actions in the large intestine. However, there is little known of the role VDR plays in regulating colonic epithelial transport function. Interestingly, previous studies from our laboratory have shown the nuclear bile acid receptor, farnesoid X receptor (FXR), to exert anti-secretory actions in colonic epithelial cells. The current study aims to investigate a potential role for VDR in regulating colonic Cl- secretion and whether it may be a target for the treatment of diarrhoeal diseases.

AIM: To investigate the effects of VDR activation by calcitriol on colonic epithelial Cl- secretion.

METHODS: Polarised monolayers of T84 cells, grown on permeable supports, were used as a model of the colonic epithelium. Protein expression of VDR and CFTR was measured by western blotting. Calcitriol (1 - 20 nM) was used to activate VDR and the VDR target protein, CYP24A1, was used to verify activation of the receptor by qRT-PCR. The specificity of calcitriol as a VDR agonist was determined by investigating FXR activation using the FXR target protein, FGF19, and a FXR-luciferase reporter cell line. The effects of VDR activation by calcitriol (10 nM) on chloride secretion over a timecourse (6 - 48 hr) was determined using the Ussing chamber technique. Changes in short-circuit current (Isc) induced by the Ca2+-dependent agonist, carbachol (CCh; 100 µM) were assessed.

RESULTS: VDR was found to be expressed in T84 cell monolayers as determined by western blotting. Treatment of T84 cells with calcitriol (5 - 20 nM) significantly (n = 3, *p ≤ 0.05) increased expression of CYP24A1, indicating activation of VDR by the agonist. Calcitriol did not alter the activity of FXR, either in an FXR-luciferase reporter cell line or in T84 cells, indicating specificity of the agonist for VDR. In Ussing chambers, the FXR agonist, GW4064 (5 µM; 24 hrs), inhibited Cl- secretory responses to CCh, as previously reported (n = 5, **p ≤ 0.01). Calcitriol (10 nM) was also found to significantly inhibit CCh-stimulated Cl- secretory responses by 39.9 ± 5.1% (6 hr), 40.4 ± 4.3% (18 hr) and 62.0 ± 6.1% (24 hr) (n = 5, **p ≤ 0.01). In addition to their effects on CCh-induced chloride secretion, GW4064 (5 µM; 24 hrs) downregulated the protein expression of CFTR by 0.5 ± 0.1 (n = 8, ***p ≤ 0.001) (as previously reported) and calcitriol (10 nM) downregulated CFTR by 0.35 ± 0.1 (n=8, *p≤ 0.05).

CONCLUSION: Similar to the bile acid receptor, FXR, VDR is expressed in colonic epithelial cells where activation by its natural ligand, calcitriol, downregulates CFTR expression and inhibits agonist-induced Cl- secretory responses. Since Cl- secretion is the primary driving force for intestinal fluid secretion, our data suggest that VDR may serve as a target for the development of new anti-diarrhoeal agents. Future studies will aim to determine the molecular mechanisms involved and investigate a possible role for VDR in mediating anti-secretory actions of luminal bile acids.

PMID:35557128 | DOI:10.1096/fasebj.2022.36.S1.R4879

J Antibiot (Tokyo). 2022 May 13. doi: 10.1038/s41429-022-00530-w. Online ahead of print.

ABSTRACT

Burkholderia cepacia complex (Bcc) species are aerobic, Gram-negative and non-fermantative bacilli. Bcc can cause clinical symptoms in patients with cystic fibrosis, ranging from asymptomatic carriage to fatal pneumonia. A pressing need exists for new antimicrobial agents that target Bcc. Ceragenins, CSA-13, CSA-131 and CSA-131 with 5% Pluronic® F127 (CSA-131P), were evaluated against Bcc clinical isolates (n = 42). MICs of ceragenins and conventional antibiotics were determined. Time-kill curve experiments were performed with 1x, 4x MICs of ceragenins and sulfamethoxazole-trimethoprim (SXT), levofloxacin. MIC50/ MIC90 results (mg l-1) of CSA-13, CSA-131 and CSA-131P were determined as 16/64, 16/128 and 16/128, respectively. CSA-13 and CSA-131 showed bactericidal activity. CSA-13 - levofloxacin combination displayed synergistic activity against Bcc. First-generation (CSA-13) and second-generation (CSA-131 and CSA-131P) ceragenins have significant antimicrobial effects on Bcc. The findings of this study demonstrate that combinations of ceragenins with currently marketed antibiotics could be synergistic in vitro against Bcc isolates. These results suggest that combination therapy with conventional antibiotics could be an alternative approach for treating Bcc infections in the future.

PMID:35562592 | DOI:10.1038/s41429-022-00530-w

Respir Physiol Neurobiol. 2022 May 10:103919. doi: 10.1016/j.resp.2022.103919. Online ahead of print.

ABSTRACT

BACKGROUND: Indices of ventilation heterogeneity (VH) from multiple breath washout (MBW) have been shown to correlate well with VH indices derived from hyperpolarised gas ventilation MRI. Here we report the prediction of ventilation distributions from MBW data using a mathematical model, and the comparison of these predictions with imaging data.

METHODS: We developed computer simulations of the ventilation distribution in the lungs to model MBW measurement with 3 parameters: σV, determining the extent of VH; V0, the lung volume; and VD, the dead-space volume. These were inferred for each individual from supine MBW data recorded from 25 patients with cystic fibrosis (CF) using approximate Bayesian computation. The fitted models were used to predict the distribution of gas imaged by 3He ventilation MRI measurements collected from the same visit.

RESULTS: The MRI indices measured (I1/3, the fraction of pixels below one-third of the mean intensity and ICV, the coefficient of variation of pixel intensity) correlated strongly with those predicted by the MBW model fits (r=0.93,0.88 respectively). There was also good agreement between predicted and measured MRI indices (mean bias ± limits of agreement: I1/3:-0.003±0.118 and ICV:-0.004±0.298). Fitted model parameters were robust to truncation of MBW data.

CONCLUSION: We have shown that the ventilation distribution in the lung can be inferred from an MBW signal, and verified this using ventilation MRI. The Bayesian method employed extracts this information with fewer breath cycles than required for LCI, reducing acquisition time required, and gives uncertainty bounds, which are important for clinical decision making.

PMID:35562095 | DOI:10.1016/j.resp.2022.103919

Int J Infect Dis. 2022 May 10:S1201-9712(22)00274-0. doi: 10.1016/j.ijid.2022.05.012. Online ahead of print.

ABSTRACT

OBJECTIVES: Burkholderia gladioli has been associated with infections in patients with cystic fibrosis, chronic granulomatous disease and other immunocompromising conditions. The aim of this study was to better depict the outbreak of healthcare-associated bacteremia caused by Burkholderia gladioli due to contaminated multi-dose vials with saline solutions.

METHODS: An environmental and epidemiological investigation was conducted by the Infection Prevention and Control Team (IPCT) in order to identify the source of the outbreak in three Croatian hospitals.

RESULTS: During a three-month period, a total of thirteen B. gladioli bacteremia episodes were identified in ten patients in three Croatian hospitals. At the time of the outbreak, all three hospitals used saline products from the same manufacturer. Two 100 ml multi-dose vials with saline solutions and needleless dispensing pins were B. gladioli positive. All thirteen bacteremia isolates, as well as two isolates from the saline, showed the same antimicrobial susceptibility patterns and pulsed-field gel electrophoresis profile demonstrating clonal relatedness.

CONCLUSIONS: When an environmental pathogen causes an outbreak, contamination of intravenous products must be considered. Close communication between local IPCT and the National Hospital Infection Control Advisory Committee is essential to conduct a prompt and thorough investigation and find the source of the outbreak.

PMID:35562041 | DOI:10.1016/j.ijid.2022.05.012

FASEB J. 2022 May;36 Suppl 1. doi: 10.1096/fasebj.2022.36.S1.R2035.

ABSTRACT

Based on the CREST (Connecting Researchers, Educators, and STudents) model undergraduate students at Nova Southeastern University (NSU) cooperated in teams through a specially designed Honors Seminar Course titled, "Introduction to Protein Modeling". This Course-based Undergraduate Research Experience (CURE) brought together groups of undergraduate students early in their studies to have authentic research and presentation experiences. This course was open to all students in NSU's Farquhar Honors College regardless of their major, academic level or previous college level experience with science or protein modeling. Out of 15 registered students, seven of them were first semester freshmen, one was a sophomore, three were juniors, and four were seniors. Most of the students were Biology or Behavioral Neuroscience Majors. However, there was one of each of the following four majors: Dance, Exercise and Sport Science, Marine Biology, and Pre-Nursing. The students were divided into five groups of three, each consisting of a mix of freshmen and upper-level students. Here we present the design and results of a course that provided students with early access to a course-based research experience using various protein bioinformatics tools (Jmol, Pymol, Autodoc Vina, and the Protein Data Bank). Students were shown previously completed projects as part of the CREST Program available on the Program's website: https://cbm.msoe.edu/crest/new9_2021.php. To support CURE sustainability, we used an interdisciplinary team-teaching approach, with distributed leadership and the support of the University's Honors College in the development of this course. Students worked collaboratively during class meetings to identify a protein molecular story that they wanted to research and describe using molecular modeling tools, including the development of a 3-D model. At the midterm, the student groups shared their progress and received feedback from the faculty and their peers. Students continued to revise their modeling projects and received additional feedback one month later. At the end of the semester each team presented their final projects with components that included a protein model description sheet, poster, and oral presentations. Student projects included molecular stories related to MMP 12, cystic fibrosis transmembrane regulator, marine bacterial laminarinase, sickle cell anemia, and iron acquisition in E. coli related to TonB/TonBox binding. Through interacting with the primary literature, course materials, and using protein modeling tools, students gained an introduction to the scientific process and applied it to better understand molecular mechanisms. During this process, students also learned to explain these processes to themselves and others through constructing presentations and model descriptions. Student learning gains were documented by using the RISC (Research on the Integrated Science Curriculum) Survey.

PMID:35560647 | DOI:10.1096/fasebj.2022.36.S1.R2035

Am Fam Physician. 2022 May 1;105(5):469-478.

ABSTRACT

Constipation in children is usually functional constipation without an organic cause. Organic causes of constipation in children, which include Hirschsprung disease, cystic fibrosis, and spinal cord abnormalities, commonly present with red flag signs and symptoms. A history and physical examination can diagnose functional constipation using the Rome IV diagnostic criteria. The first goal of managing constipation is to treat fecal impaction, and then maintenance therapy is used to prevent a recurrence. Polyethylene glycol is the first-line treatment for constipation. Second-line options include lactulose and enemas. Increasing dietary fiber and fluid intake above usual daily recommendations and adding probiotics provide no additional benefits for treating constipation. Frequent follow-up visits and referrals to a psychologist can assist in reaching some treatment goals. Clinicians should educate caregivers about the chronic course of functional constipation, frequent relapses, and the potential for prolonged therapy. Clinicians should acknowledge caregivers' specific challenges and the negative effects of constipation on the child's quality of life. Referral to a pediatric gastroenterologist is recommended when there is a concern for organic causes or constipation persists despite adequate therapy.

PMID:35559625

FASEB J. 2022 May;36 Suppl 1. doi: 10.1096/fasebj.2022.36.S1.R3279.

ABSTRACT

The Carbohydrate sulfotransferase (CHST) family is essential for Chondroitin Sulfate synthesis and is associated with cancer progression. Comparing all CHST families, we found that CHST11 has prognostic value than other families. CHST11 is related to the overall survival rate in the pan-cancer profile, among which lung adenocarcinoma is the most significant. From pulmonary fibrosis to lung cancer, the expression of CHST11 showed a linear increase. We analyzed the CHST11 interactome, and the results indicated that CHST11 contributes to diverse canonical pathways, including immune microenvironment, fibrosis, and cancer progression. Based on the prediction results of Ingenuity Pathway Analysis (IPA), we speculate that CHST11 can secrete IL1B and IL6 signals to activate the upstream regulator IRF8. This is also the key to the fact that IRF8 has contributed to cystic fibrosis and lung cancer progression. In addition, IRF8 can also transactivate CD80 and CD86 to reform the tumor microenvironments and immune response. By reversing the CHST11-IL6/IL1B-IRF8-CD80/CD86 mediated axis, we confirmed that VE821 and LY2603618 have the value of drug repurposing. Our study proves novel insight into how CHST11 contributes to cystic fibrosis and lung cancer by reprogramming the immune microenvironment.

PMID:35559534 | DOI:10.1096/fasebj.2022.36.S1.R3279

J Pediatr Pharmacol Ther. 2022;27(4):396-399. doi: 10.5863/1551-6776-27.4.396. Epub 2022 May 9.

ABSTRACT

This report describes a case of a 15-year-old male with cystic fibrosis caused by N1303K and Q493X cystic fibrosis transmembrane conductance regulator (CFTR) protein variants. In this case, CFTR modulators including tezacaftor-ivacaftor and subsequently elexacaftor-tezacaftor-ivacaftor were used and resulted in clinical stability and improvement.

PMID:35558347 | PMC:PMC9088447 | DOI:10.5863/1551-6776-27.4.396

J Pediatr Pharmacol Ther. 2022;27(4):306-311. doi: 10.5863/1551-6776-27.4.306. Epub 2022 May 9.

ABSTRACT

OBJECTIVE: Vitamin D deficiency is commonly found in patients with cystic fibrosis (CF) and can have a negative effect on patients who are not at target goal according to Cystic Fibrosis Foundation's Vitamin D Deficiency Clinical Care Guidelines. The objective of this study is to determine the effectiveness of a pharmacist-driven vitamin D protocol (PDVDP) in improving, achieving, and maintaining 25-hydroxyvitamin D levels of patients in a pediatric CF clinic.

METHODS: A retrospective chart review was conducted for pediatric patients with CF from August 2018 to March 2020 to determine the percent of patients with improvement in 25-hydroxyvitamin D levels to target goal (≥ 30 ng/mL). Patients' 25-hydroxyvitamin D levels at 6, 12, and 18 months after automatic enrollment into the PDVDP were compared to determine if improvement occurred, and to calculate relative percent increase of 25-hydroxyvitamin D levels for these patients.

RESULTS: The mean 25-hydroxyvitamin D levels of the patients at baseline before the protocol and 6, 12, and 18 months after enrollment in the protocol were 23.2, 33.3, 32.7, and 34.6 ng/mL, respectively. These results demonstrate mean 25-hydroxyvitamin D levels at all follow-up time points were significantly greater than baseline (p < 0.001). At 6 months, 50% (n = 20) of pediatric patients reached the target 25-hydroxyvitamin D levels.

CONCLUSIONS: The PDVDP was effective in increasing the number of patients able to reach target 25-hydroxyvitamin D levels. Our PDVDP process may also be used at other CF clinics to improve vitamin D outcomes collaboratively with the interprofessional CF team.

PMID:35558342 | PMC:PMC9088445 | DOI:10.5863/1551-6776-27.4.306

FASEB J. 2022 May;36 Suppl 1. doi: 10.1096/fasebj.2022.36.S1.L7676.

ABSTRACT

Relevant to multiple studies, calcium activated chloride channel called Anoctamin1 (ANO1) is identified as an excellent biomarker for tumor growth reported in different cancer cells such as prostate, breast, and lung cancer. Growing evidence suggests, ANO1 promotes tumor growth via regulating EGFR signaling and modulating MAPK and protein kinase B pathway. In non-small cell lung cancer (NSCLC), the ANO1 expression level is high, leading to high cancer-related mortality. To date, several natural and synthetic compounds have shown their modulatory effect on ANO1 activity but are out of the limelight due to selectivity. Our present study is carried out based on finding a selective and potent modulator of ANO1 activity by in silico and in vitro assay. According to the hypothesis, inhibitors of ANO1 are screened out, where an inhibitor reduced ANO1 expression without modulating intracellular Ca+ level and cystic fibrosis transmembrane conductance regulator (CFTR) channel activity. The molecular docking study revealed negative high binding free energy than known inhibitor Ani9 and other candidate compounds. Furthermore, our potential compound suppressed cell viability and migration process in ANO1 expressing PC9 cells, while not in H1975 cells not expressing ANO1. Besides, it initiated the apoptosis process by up-regulating caspase-3 activity. Thus, the above results provided significant evidence of the anticancer effect of the final compound against ANO1, reducing its protein level in NSCLC.

PMID:35557374 | DOI:10.1096/fasebj.2022.36.S1.L7676

FASEB J. 2022 May;36 Suppl 1. doi: 10.1096/fasebj.2022.36.S1.R3684.

ABSTRACT

Mummichogs (Fundulus heteroclitus) can inhabit a wide range of environmental salinities because of their capacity to modulate the expression of genes that encode solute transporters and water channels within key osmoregulatory organs. Over 60 years ago, the pituitary hormone, prolactin (Prl), was identified as an essential factor that enables mummichogs to tolerate freshwater (FW) environments; nonetheless, the molecular targets of Prl signaling in this model teleost remain unknown. In the current study, we determined that two Prl receptors, designated prlra and prlrb, are expressed in osmoregulatory organs in salinity-dependent fashions. We administered ovine Prl (oPrl; 1 or 5 mg g-1 body weight) by intraperitoneal injection to mummichogs held in brackish water (12 ppt) and first characterized the expression of genes associated with FW-type (ion absorptive) and seawater (SW)-type (ion secretory) ionocytes. Within FW-type ionocytes, oPrl stimulated the expression of Na+ /Cl- cotransporter 2, aquaporin 3, and prlra. Branchial Na+ /H+ exchanger 2and -3 gene expression did not respond to oPrl. The expression levels of genes associated with SW-type ionocytes, such as Na+ /K+ /2Cl- cotransporter 1and cystic fibrosis transmembrane regulator 1, were reduced following oPrl administration. Given the role of aquaporins in supporting fluid absorption by the intestine, we then assessed whether multiple aquaporin encoding genes were responsive to oPrl. We observed that aquaporin 1a and -8 were diminished by oPrl. As in the gill, oPrl stimulated the expression of prlra. Our collective data indicate that Prl promotes the survival of mummichogs in FW by simultaneously modulating a suite of genes within the gill and intestine that underlie critical ion- and water-transporting processes.

PMID:35557127 | DOI:10.1096/fasebj.2022.36.S1.R3684

FASEB J. 2022 May;36 Suppl 1. doi: 10.1096/fasebj.2022.36.S1.R4152.

ABSTRACT

Type 4 cyclic nucleotide phosphodiesterases (PDE4s) comprise a family of four isoenzymes, PDE4A to D, that hydrolyze the second messenger cAMP. Three of these enzymes, PDE4A, PDE4B and PDE4D, are widely expressed throughout pulmonary and immune cell types, and their concurrent inhibition with non/PAN-selective PDE4 inhibitors is well-known to exert potent anti-inflammatory benefits. To determine if PDE4 inhibition may represent a therapeutic approach for acute lung injury resulting from severe pneumonia, we explored the effect of genetic ablation of individual PDE4 subtypes, or their concurrent inhibition with PAN-PDE4 inhibitors, in a model of acute intra-tracheal Pseudomonas aeruginosa (PA) infection in mice. We have shown recently that selective ablation of PDE4B is sufficient to protect mice from lung injury in this model by reducing pulmonary and systemic levels of pro-inflammatory cytokines, as well as pulmonary vascular leakage and mortality. With the present study we explored whether selective inactivation of PDE4A or treatment with PAN-PDE4 inhibitors may provide additional therapeutic benefits.

METHODS: Mice deficient in PDE4A (PDE4A-KO) and their wildtype (WT) littermate controls were infected intratracheally with the PA lab strain PA01 and subjected 16 h later to bronchoalveolar lavage (BAL), cardiac blood draws, and tissue extractions. Alternatively, vascular/alveolar leakage was assessed by measuring Evans Blue Dye extravasation. To test the effect of PAN-PDE4 inhibition, mice were treated with the PDE4 inhibitor Piclamilast (5 mg/kg, i.p.) at 2 h prior to PA01 infection.

RESULTS: Compared to WT controls, PDE4A-KO mice displayed a hyper-inflammatory phenotype characterized by increased levels of pro-inflammatory cytokines, immune cells (neutrophils) in BAL fluid, and vascular leakage. Despite these hyperinflammatory responses, the bacterial load in lungs was also increased in PDE4A-KO mice compared to WT controls. Treatment with a PAN-PDE4 inhibitor produced incongruent effects, with an increased bacterial load mirroring the effect of PDE4A ablation, whereas reduced levels of inflammatory cytokines such as TNFα mimicked the effect of PDE4B ablation.

CONCLUSIONS: Ablation of PDE4A produces detrimental effects in this model of acute PA-lung infection. Current studies aim to determine whether an impaired bacterial killing, and the resulting increase in bacterial load, are responsible for the hyperinflammation, or whether these are independent effects of PDE4A ablation. PAN-PDE4 inhibition mimics some of the beneficial effects of selective PDE4B ablation, but also some of the detrimental effects of PDE4A ablation. These findings suggest that selective inactivation of PDE4B may be a more effective therapeutic approach, compared to PAN-PDE4 inhibition, in settings of acute PA-lung infection.

PMID:35556782 | DOI:10.1096/fasebj.2022.36.S1.R4152

FASEB J. 2022 May;36 Suppl 1. doi: 10.1096/fasebj.2022.36.S1.R6139.

ABSTRACT

BACKGROUND-: Cystic Fibrosis (CF) is an inherited genetic disorder resulting from mutations in the gene encoding cystic fibrosis transmembrane conductance regulator (CFTR). For reasons that are not completely clear, CF patients over the age of 40 are 4-8 times more likely to develop colon cancer than the general population. Moreover, positive history of distal intestinal obstruction syndrome or organ transplant further increase risk of developing colon cancer by 11-fold.

OBJECTIVE-: The objective of this research is to test the hypothesis that inherited CFTR mutations in CF patients promote colon cancer development by increasing cell migration and proliferation via altered epidermal growth factor (EGFR) signaling.

METHODS-: Scratch assays were performed on confluent fetal human colon epithelial cells (FHC) and the rate of cell migration was measured 1-5 days after treatment with either CFTR activating (IBMX and forskolin) or inhibiting compounds (CFTRinh -172 and CFTR siRNA). Ki67 nuclear antigen and EGFR labeling were conducted using standard immuno-histochemical labeling and detection techniques.

RESULTS-: We found that both pharmacological and siRNA inhibition of CFTR activity increased the rate of wound closure and cell migration. Conversely, IBMX and forskolin stimulation of CFTR decreased the rate of FHC migration (vs untreated control rates). Inhibition of CFTR also associated with increased levels of Ki67 (a classic cell proliferation marker) and increased expression of EGFR (which has known roles in cell growth and differentiation).

CONCLUSION-: Our results indicate that CFTR plays an important role in FHC colon cell migration and EGFR signaling, which could lead to a better understanding as to why CF patients have increased risk for colon cancer.

PMID:35556394 | DOI:10.1096/fasebj.2022.36.S1.R6139