Crit Rev Clin Lab Sci. 2025 Jul 17:1-11. doi: 10.1080/10408363.2025.2527288. Online ahead of print.

ABSTRACT

Over the past 60 years, preventative public health screening programs have evolved since their inception and now include newborn screening (NBS) aimed at identifying infants after birth for a number of rare, congenital, inherited diseases. Most of the conditions detected through NBS are autosomal recessive disorders or exhibit X-linked inheritance, meaning that family members of individuals with these conditions have a higher risk for being either affected or obligate heterozygotes. For example, the X-linked adrenoleukodystrophy (X-ALD) in the screening panel identifies affected newborns and asymptomatic relatives through subsequent testing. Thus, NBS becomes a gateway to family-wide prevention, through the application of reverse cascade testing (RCS). In this paper we examined the scenarios where RCS may be appropriate. Accordingly, we have identified a list of criteria assessing whether a NBS disease would benefit from RCS: (1) autosomal recessive or X-linked inheritance; (2) high carrier rates, (3) variable expressivity, (4) mild or late-onset forms; and (5) association with diagnostic delays and recent addition to the screening panel. More than one criterion usually needs to be met for a disease to benefit from RCS. We have identified a list of diseases and highlighted the potential benefits of RCS: X-ALD, Cystic Fibrosis, Sickle Cell Disease, Spinal Muscular Atrophy and Pompe disease. There are additional scenarios within NBS where disease maternal conditions (3-methylcrotonyl-CoA carboxylase deficiency and carnitine uptake deficiency) or nutritional maternal conditions (vitamin B12 deficiency) may cause a screen-positive NBS result. Whenever a maternal nutritional deficiency is a potential reason for a positive NBS, this is indicative of a non-inherited condition that may require treatment in the newborn owing to possible neurological damage and delay in normal growth in newborns with certain secondary deficiencies. For these cases RCS is recommended, as the mother's status may put her at risk for future adverse events (i.e. cardiovascular and musculoskeletal disorders, hepatic involvement, and neurodegeneration). The RCS-NBS strategy discussed in this paper offers a set of criteria against which diseases can be assessed for the potential need for RCS. Implementation of this strategy requires several considerations including educational needs, ethical issues, uptake of testing, logistics and costs for this expanded screening and counseling, and availability of appropriate specialists for ongoing management.

PMID:40673334 | DOI:10.1080/10408363.2025.2527288

J Clin Transl Endocrinol. 2025 Jun 23;41:100403. doi: 10.1016/j.jcte.2025.100403. eCollection 2025 Sep.

ABSTRACT

Cystic fibrosis-related diabetes (CFRD) is the most prevalent extrapulmonary comorbidity in CF. While insulin remains the standard treatment, increasing rates of overweight and obesity due to CFTR modulators highlight the need for new therapies. SGLT2 inhibitors, effective in type 2 diabetes, have not been studied in CFRD. This case series presents eight CFRD patients treated with SGLT2 inhibitors alongside insulin for one year. Half had BMI reductions (1.33-2.89 kg/m2); others had increases. Glycemic control improved in six, worsened in two due to insulin nonadherence. Insulin adjustments showed no pattern. One patient discontinued due to genital infections; overall tolerance was high.

PMID:40672774 | PMC:PMC12264597 | DOI:10.1016/j.jcte.2025.100403

bioRxiv [Preprint]. 2025 Jul 8:2025.07.08.663034. doi: 10.1101/2025.07.08.663034.

ABSTRACT

Acute respiratory viral infections are an important driver of morbidity and mortality in people with chronic lung disease and are frequently associated with pulmonary exacerbations and a transition from intermittent to chronic bacterial infection of the airways. Chronic Pseudomonas aeruginosa infections are associated with worsened lung function, poor outcomes, and increased hospital visits. We sought to improve understanding of the effects of respiratory viral infections and host immune response on the resident bacterial community of the airways, using cystic fibrosis as a model. We performed an observational longitudinal study of 38 adults with CF and collected sinus and sputum samples at 6-month intervals from 2017 - 2021. We performed 16S rRNA amplicon sequencing to characterize the airway microbiota, real-time RT-PCR for viral infection detection, and cytokine quantification. We observed viral positivity rates of 19% and 14% in sinus and sputum samples, respectively. Human rhinovirus was the most frequently observed viral pathogen in both sinus and sputum samples. We measured a significant perturbance of the bacterial community during viral infection that did not return to baseline following resolution of the viral infection. This perturbation was driven by a significant increase in Pseudomonas relative abundance during viral infection. Furthermore, we found significant associations with increased Pseudomonas relative abundance for several pro-inflammatory and antiviral cytokines, including interleukin (IL)-2, IL-8, and interferon (IFN)- λ 1. These findings indicate an important role for respiratory viral infections and the host immune response in the development and maintenance of chronic Pseudomonas infections in the context of CF airway disease and broadly expand our understanding of viral-bacterial coinfection of the airways.

IMPORTANCE: Respiratory infections are a leading cause of morbidity and mortality worldwide, and co-infections are associated with worsened disease outcomes. In viral-bacterial co-infection, clinical and mechanistic studies show that a preceding acute respiratory viral infection promotes the establishment and exacerbation of bacterial infections, leading to increased morbidity. Although people with cystic fibrosis do not experience more frequent acute respiratory viral infections, their outcomes are worse, with prolonged symptoms and hospitalizations. When examining how acute viral infections shape the microbiota in the respiratory tract of pwCF, we observe a disturbance of the microbial community composition during viral infections that does not return to baseline after the acute viral infection resolves. Moreover, we show that Pseudomonas relative abundance is significantly increased in the airways of pwCF during viral infection and that increased concentrations of antiviral cytokines - such as interferon (IFN)- λ 1 - are associated with increased Pseudomonas abundance. These findings offer evidence that the progression of chronic Pseudomonas infections in pwCF are influenced by acute respiratory viral infections and the subsequent antiviral response in the airways. This study furthers our understanding of viral-bacterial coinfection in the context of CF.

PMID:40672159 | PMC:PMC12265526 | DOI:10.1101/2025.07.08.663034

Pediatr Pulmonol. 2025 Jul;60(7):e71210. doi: 10.1002/ppul.71210.

ABSTRACT

BACKGROUND: Cystic Fibrosis Bone Disease (CFBD) is a known complication in children with CF and may cause serious problems in adulthood or transplantation processes. This study aimed to identify potential predictable risk factors for the development of low BMD by evaluating pediatric patients screened with DXA as a "Heart-Lung Transplantation Center" and created new strategic plans to improve our CFBD screening program by evaluating our results in literature and guidelines recommendations.

METHODS: This retrospective cohort study includes 86 children ages 6-18 years with CF who underwent at least one DXA scan between August 2016 and October 2024. Participants were compared according to BMD z scores and the relationship between BMD and disease-related parameters was evaluated.

RESULTS: The rate of DXA screening in our center was 81.1% over 8 years of age and 72.8% over 6 years of age. 41.8% of our population had abnormal BMD (z scores < -1), and the rate of very low BMD (z scores < -2) was 17.4%. The frequency of abnormal BMD was higher in children with BMI< 50th percentile, SKS ≤ 70, low FEV1 z score, respiratory microorganism colonization, ≥ 2 annual pulmonary exacerbations, required respiratory support, low albumin, and high CRP levels. Systemic inflammation marker CRP increase was the most predictable parameter for low BMD.

CONCLUSION: This study informs clinical practice by highlighting the need for multidisciplinary interventions, such as earlier evaluation of DXA scans due to the risk factors and poor clinical conditions, a consistent follow-up protocol, individualized nutrition programs with the dietitian, and enhanced physical therapy.

PMID:40671433 | DOI:10.1002/ppul.71210

Pediatr Pulmonol. 2025 Jul;60(7):e71200. doi: 10.1002/ppul.71200.

ABSTRACT

BACKGROUND: With routine newborn screening for cystic fibrosis (CF) now considered standard of care, the designation of CFTR Related Metabolic Syndrome (CRMS) or CF screen positive, indeterminate diagnosis (CFSPID) has been established. The majority of CRMS/CFSPID patients remain asymptomatic; however, 3.8%-44% of these patients may progress to a diagnosis of CF. This raises the question of how to optimally manage CRMS/CFSPID patients. We set out to gain a better understanding of the past practices employed at CF centers across Canada in the care of patients with a diagnosis of CRMS/CFSPID.

METHODS: An invitation to participate in an online survey was disseminated to CF centers in Canada through the REDCap database. The survey was completed in 2018. It included questions addressing patient population, timing of follow-up of CRMS/CFSPID patients, and details around specific investigations ordered.

RESULTS: Twelve out of 20 qualifying clinics completed the survey. The total patient population compiled included 1412 patients, of which 171 (12%) were classified as CRMS/CFSPID. There was wide variability in the timing of follow-up with a median (IQR) of 6 (5.25-12) months and a range of 3-12 months. There was also wide variability in the timing of repeat investigations such as sweat chloride, respiratory cultures, chest x-rays and spirometry.

CONCLUSIONS: With current evidence showing that a considerable number of CRMS/CFSPID patients may progress to CF, ensuring these patients are identified as early as possible and followed in a consistent manner is essential. The Cystic Fibrosis Foundation and European Cystic Fibrosis Society have recently developed guidelines regarding the care of these patients. This survey describes historical practices for follow up of CRMS/CFSPID patients to help inform the development of Canadian consensus guidelines.

PMID:40671429 | DOI:10.1002/ppul.71200

Microbes Infect. 2025 Jul 14:105546. doi: 10.1016/j.micinf.2025.105546. Online ahead of print.

ABSTRACT

Staphylococcus aureus infections remain a great concern in people with cystic fibrosis also after the introduction of modulator therapy. Here we describe the state of the art of traditional and novel therapeutic strategies to fight both acute and chronic infections caused by sensitive and drug resistant strains.

PMID:40669745 | DOI:10.1016/j.micinf.2025.105546

bioRxiv [Preprint]. 2025 Jun 24:2025.06.18.660463. doi: 10.1101/2025.06.18.660463.

ABSTRACT

Cystic Fibrosis (CF) is caused by loss-of-function mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR), an anion channel predominantly expressed on the apical membrane of epithelial cells. Reduced Cl - and HCO 3 - secretion due to dysfunctional CFTR results in a decrease in lung function and is the leading cause of morbidity in individuals with CF. Recent therapies, known as highly effective CFTR modulator therapy (HEMT), help improve the lung function in individuals with specific CF-causing mutations by enhancing the folding, trafficking, and gating of CFTR. However, variability in HEMT responsiveness leads to sub-optimal clinical outcomes in some people with CF undergoing modulator therapy. A potential strategy is to complement their function with a CFTR-independent mechanism. One possibility is the use of ion channel-forming small molecules such as amphotericin B, which has shown promise in restoring function and host defenses in CF airway disease models. Amphotericin B functions as a molecular prosthetic for CFTR and may thus complement CFTR modulators. Here we show that co-treatment of CF airway epithelia with HEMT and amphotericin B results in greater increases in both HCO 3 - secretory flux and ASL pH compared to treatment with either agent alone. These findings suggest that co- administration of CFTR modulators and molecular prosthetics may provide additive therapeutic benefits for individuals with CF.

PMID:40667211 | PMC:PMC12262498 | DOI:10.1101/2025.06.18.660463

Glycoconj J. 2025 Jul 15. doi: 10.1007/s10719-025-10191-0. Online ahead of print.

ABSTRACT

Cystic Fibrosis (CF), a life-threatening hereditary disease, arises from mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene, which encodes a chloride-conducting channel widely expressed in epithelial cells. The most common mutation, F508del, causes CFTR misfolding, premature degradation, and impaired mucociliary clearance, leading to recurrent respiratory infections and inflammation. The triple combination therapy with Elexacaftor, Tezacaftor, and Ivacaftor (ETI) has revolutionized CF management by partially restoring mutated CFTR function. However, enhancing CFTR rescue and stabilizing host immune responses remain critical challenges. In airway epithelial cells, CFTR interacts with proteins and lipids in macromolecular complexes that influence its stability. Among these, the ganglioside GM1 plays a key role in modulating plasma membrane protein dynamics, including CFTR. This study investigates the effects of exogenous GM1 supplementation as an adjuvant to ETI treatment. Our results demonstrate that GM1 enhances F508del-CFTR maturation and stability, even under Pseudomonas aeruginosa infection, which typically suppresses CFTR expression and function. Furthermore, GM1 restores xenophagic activity in bronchial epithelial cells, improving host defence mechanisms against the bacteria. These findings underscore the therapeutic potential of GM1 and its analogues in optimizing the plasma membrane environment for CFTR correction, suggesting that by enhancing the efficacy of CFTR modulators, GM1 could pave the way for innovative approaches to improve CF management.

PMID:40663265 | DOI:10.1007/s10719-025-10191-0

Expert Rev Respir Med. 2025 Jul 15. doi: 10.1080/17476348.2025.2535764. Online ahead of print.

ABSTRACT

INTRODUCTION: Vitamin D deficiency is common in cystic fibrosis (CF) and may be linked to disease severity. We aimed to investigate the association between vitamin D deficiency and severity in non-CF bronchiectasis.

METHODS: A systematic search of PubMed and Scopus (up to December 2024) identified relevant studies. After screening 170 articles, seven met the inclusion criteria. Study quality was assessed using NIH tools.

RESULTS: Patients with non-CF bronchiectasis had significantly lower serum 25-hydroxyvitamin D (25OHD) levels compared to healthy controls. In one study, median 25OHD was 24.7 nmol/L in patients vs. 45.3 nmol/L in controls. Another reported mean levels of 14.7 ± 9.6 ng/mL vs. 19.8 ± 6.9 ng/mL, respectively. Disease severity was assessed using validated and semi-structured measures, including the bronchiectasis severity index (BSI), number of exacerbations, pulmonary function tests (PFTs), radiological scores (Bhalla, modified Reiff), and health-related quality-of-life (HRQL) tools. Most studies reported worse severity outcomes in vitamin D-deficient patients.

CONCLUSIONS: Although vitamin D deficiency appears to be associated with more severe non-CF bronchiectasis, heterogeneity between studies limits definitive conclusions. Future studies should incorporate standardized tools such as the eFACED score to better characterize disease severity.

PMID:40662885 | DOI:10.1080/17476348.2025.2535764

Pediatr Pulmonol. 2025 Jul;60(7):e71208. doi: 10.1002/ppul.71208.

NO ABSTRACT

PMID:40662499 | DOI:10.1002/ppul.71208

Pediatr Pulmonol. 2025 Jul;60(7):e71190. doi: 10.1002/ppul.71190.

ABSTRACT

BACKGROUND: The HERO-2 study is a prospective, observational study investigating chronic daily therapy (CDT) changes among people with cystic fibrosis (PwCF) ≥ 12 years of age taking elexacaftor/tezacaftor/ivacaftor (ETI). The goal of this analysis was to describe baseline characteristics and patterns of CDT discontinuation reported at study enrollment and identify clinical features associated with CDT discontinuation.

METHODS: Study participants were recruited through their CF center, community engagement, or a smartphone application (Folia Health). Study enrollment and participation took place remotely through Folia Health. Participants gave consent to link their data with the CF Foundation Patient Registry (CFFPR). At enrollment, participants completed a baseline survey describing CDT use since starting ETI. Descriptive statistics were used to summarize HERO-2 participant characteristics. Logistic regression evaluated the association of selected individual characteristics with odds of CDT discontinuation.

RESULTS: A total of 860 PwCF enrolled in HERO-2, and 755 (87.8%) completed the enrollment survey and were linked to the CFFPR. At enrollment, 313 participants (41.5%) reported discontinuation of at least one CDT. The most frequently discontinued CDT was airway clearance (22.4%). Intravenous (IV) antibiotic treatment for a pulmonary exacerbation in the prior year and public insurance were associated with lower odds of CDT discontinuation.

CONCLUSIONS: In this large cohort of PwCF taking ETI, approximately 42% reported discontinuation of at least one CDT. Providers should be aware of these CDT discontinuation patterns and engage in open dialogue with PwCF and their families about CDT de-escalation.

PMID:40662498 | DOI:10.1002/ppul.71190

ERJ Open Res. 2025 Jul 14;11(4):00113-2025. doi: 10.1183/23120541.00113-2025. eCollection 2025 Jul.

ABSTRACT

Bronchiectasis pathogens in Taiwan have implications on clinical outcomes https://bit.ly/4iCDeK6.

PMID:40661932 | PMC:PMC12257154 | DOI:10.1183/23120541.00113-2025

Can Respir J. 2025 Jul 1;2025:8893074. doi: 10.1155/carj/8893074. eCollection 2025.

ABSTRACT

Introduction: The Canadian Cystic Fibrosis Registry (CCFR) was developed in the 1970s and has longitudinal demographic and clinical data on persons living with cystic fibrosis (CF) attending accredited clinics in Canada. We aimed to validate the data collection and identify potential limitations of the CCFR. Methods: Of 40 accredited CF clinics in Canada invited and based on an a priori sample size calculation, eight clinics were included. 15% of each CF clinic's population in 2019 were randomly selected. Data variables were selected based on their importance to care, epidemiologic trends, and data related to demography, clinic visits, and hospitalizations. The accuracy of the registry data was compared to the medical records as the gold standard. Each data element was categorized as correct, incorrect, or not able to be validated. The accuracy rate was calculated as the percent correct out of all records validated. Results: A total of 4382 individuals had data entered into the CCFR in 2019. The validation cohort consisted of 208 individuals from 8 clinics, which were representative across location, size of clinic (small/medium/large), and type of clinic (adult, pediatric, and combined). The 208 individuals were 52% male and 95% White, and with a median age of 26.3 years (IQR: 15.2-36.6). Approximately 95% of CCFR data on clinical measurements, infections, treatments, and hospitalizations validated were accurate as compared to the medical record. For demography, sex and date of birth had 100% accuracy. Conclusion: Our validation of the CCFR demonstrated high accuracy for clinical and demographic variables used in clinical research.

PMID:40661671 | PMC:PMC12259333 | DOI:10.1155/carj/8893074

bioRxiv [Preprint]. 2025 Jun 12:2025.06.11.659172. doi: 10.1101/2025.06.11.659172.

ABSTRACT

Cryogenic electron microscopy (cryo-EM) analysis of bacteriophages is a valuable method for deciphering virus composition and conformational plasticity. In this study, we present a high-resolution structural atlas of the Pseudomonas virus Pa223, a phage from the Bruynoghevirus genus that has recently been used in clinical cocktails for treating cystic fibrosis and non-cystic fibrosis bronchiectasis, as well as for compassionate care. By combining bioinformatics, proteomics, cryo-EM single particle analysis, and localized reconstruction, we annotated and built atomic models for eight structural polypeptide chains that form the icosahedral capsid and noncontractile tail. We discovered that the Pa223 capsid is decorated by a spike protein that features a unique triple-β helix fold with no structural homologs in the database. The Pa223 tail features six trimeric tail fibers extending upwards, similar to, but shorter than, those found in phage T7. Unlike T7, the Pa223 tail is extended by two head-to-tail adaptors and sealed by a trimeric tail needle, similar to P22-like phages. We identified a protein bound around the outer perimeter of the portal protein, positioned similarly to the ejection protein gp72, which was identified in the Pseudomonas phage DEV, a Litunavirus phage and member of the reclassified Schitoviridae family. This structural hint led us to identify the Pa223 ejection proteins gp53, gp54, and gp56, which bioinformatically resemble those of T7-like phages more closely than Schitoviridae . Thus, phage Pa223 contains diverse structural elements found in P22-like, T7-like, and Litunavirus phages, providing a framework for understanding the diversification and evolution of ejection proteins in Bruynogheviruses .

HIGHLIGHTS: The high-resolution structure of Bruynoghevirus Pa223 reveals hybrid structural features that are shared among P22-like, T7-like, and Litunavirus phages. The Pa223 capsid is decorated with a trimeric spike asymmetrically bound at the icosahedral 3-fold axes.The Pa223 tail features two quasi-equivalent conformations of the head-to-tail adaptor protein arranged into two coaxial rings. Identification of the ejection protein gp54 through structural similarity to gp72 from the Litunavirus DEV. Bioinformatic mapping of the Pa223 ejection proteins gp53 and gp56 validated through mass spectrometry analysis of infectious virions.

PMID:40661476 | PMC:PMC12259138 | DOI:10.1101/2025.06.11.659172

Nat Rev Clin Oncol. 2025 Jul 14. doi: 10.1038/s41571-025-01052-8. Online ahead of print.

ABSTRACT

Radiotherapy has an established role in the clinical treatment of patients with a variety of cancers owing to the ability to preferentially kill malignant cells mostly while sparing their non-malignant counterparts. Results from phase I-II trials also suggest that radiotherapy can have therapeutically relevant immunostimulatory effects, especially when combined with immune-checkpoint inhibitors. Over the past two decades, evidence has emerged showing that intestinal microbial communities have a major influence on the immunological tonus of patients with cancer and can influence sensitivity to various immunotherapies, including immune-checkpoint inhibitors and chimeric antigen receptor T cells. Here, we critically discuss the effects of such microbial ecosystems on radiotherapy-induced toxicities and tumour-targeting immune responses, with a focus on the clinical potential of these relationships for predictive and therapeutic clinical applications.

PMID:40659791 | DOI:10.1038/s41571-025-01052-8

Eur Respir J. 2025 Jul 14;66(1):2500793. doi: 10.1183/13993003.00793-2025. Print 2025 Jul.

NO ABSTRACT

PMID:40659466 | DOI:10.1183/13993003.00793-2025

Transfusion. 2025 Jul 14. doi: 10.1111/trf.18318. Online ahead of print.

ABSTRACT

BACKGROUND: Several studies show that extracorporeal photopheresis (ECP) might benefit chronic lung allograft dysfunction (CLAD). A standard ECP cycle consists of two consecutive procedures regardless of the technique employed.

STUDY DESIGN AND METHODS: Evaluation of ECP cycle (from two to one procedure) modification due to pandemic restrictions in 25 patients with CLAD under chronic treatment by off-line ECP in the 6 months preceding cycle modification (one procedure processing 1.5 patients blood volumes [1.5 ECP]). Assessment of any significant change in lung function decline and the relationship with product characteristics compared to pre-ECP cycle modification.

RESULTS: ECP patients (23 obstructive and two mixed) were enrolled in 2020 during the COVID pandemic. Two hundred and thirty five ECP procedures followed the standard protocol and 121 the 1.5 ECP. There was little or no variation in lung function during the study period. The mean number of mononuclear cells (MNC) per kg administered over time was higher in the 1.5 ECP than in the standard ECP protocol (p = .014). No association was found between respiratory function and MNC infused. Persistent Forced Expiratory Volume in 1 s decline >10% was observed in two patients over the 6 months preceding 1.5 ECP (due to CLAD progression) and in three patients after 1.5 ECP initiation (one for CLAD progression, two for bronchial colonization).

CONCLUSION: Our study shows that respiratory function is maintained over time and is comparable between both ECP strategies in responders. The shift from two to one procedure per cycle may be reasonable in CLAD patients treated by off-line ECP.

PMID:40657687 | DOI:10.1111/trf.18318

Bio Protoc. 2025 Jun 20;15(12):e5341. doi: 10.21769/BioProtoc.5341. eCollection 2025 Jun 20.

ABSTRACT

Epithelial tissues form barriers to the flow of ions, nutrients, waste products, bacteria, and viruses. The conventional electrophysiology measurement of transepithelial resistance (TEER/TER) can quantify epithelial barrier integrity, but does not capture all the electrical behavior of the tissue or provide insight into membrane-specific properties. Electrochemical impedance spectroscopy, in addition to measurement of TER, enables measurement of transepithelial capacitance (TEC) and a ratio of electrical time constants for the tissue, which we term the membrane ratio. This protocol describes how to perform galvanostatic electrochemical impedance spectroscopy on epithelia using commercially available cell culture inserts and chambers, detailing the apparatus, electrical signal, fitting technique, and error quantification. The measurement can be performed in under 1 min on commercially available cell culture inserts and electrophysiology chambers using instrumentation capable of galvanostatic sinusoidal signal processing (4 μA amplitude, 2 Hz to 50 kHz). All fits to the model have less than 10 Ω mean absolute error, revealing repeatable values distinct for each cell type. On representative retinal pigment (n = 3) and bronchiolar epithelial samples (n = 4), TER measurements were 500-667 Ω·cm2 and 955-1,034 Ω·cm2 (within the expected range), TEC measurements were 3.65-4.10 μF/cm2 and 1.07-1.10 μF/cm2, and membrane ratio measurements were 18-22 and 1.9-2.2, respectively. Key features • This protocol requires preexisting experience with culturing epithelial cells (such as Caco-2, RPE, and 16HBE) for a successful outcome. • Builds upon methods by Lewallen et al. [1] and Linz et al. [2], integrating commercial chambers and providing a quantitative estimate of error. • Provides code to run measurement, process data, and report error; requires access to MATLAB software, but no coding experience is necessary. • Allows for repeated measurements on the same sample. Graphical overview Electrochemical impedance spectroscopy measurement involves sending a galvanostatic signal through the electrophysiology chamber and across the epithelial cell monolayer (left) and results in complex impedance data at each frequency. This data is then fit to an electrical circuit model to output transepithelial resistance (TER), transepithelial capacitance (TEC), and membrane ratio (α) (right).

PMID:40657538 | PMC:PMC12254589 | DOI:10.21769/BioProtoc.5341

Indian J Microbiol. 2025 Jun;65(2):1209-1216. doi: 10.1007/s12088-024-01366-8. Epub 2024 Aug 13.

ABSTRACT

A microbial identification method using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF) is an innovative dimension in proteomic analysis. MALDI-ToF mass spectrometry allows not only determine the species and subspecies bacteria, but also determined by proteomic analysis and the corresponding software degree of kinship analyzed strains, which allows this method to be used in epidemiological studies and in comparing strains isolated from patients with chronic infection. The aim of this study was to evaluate the possibility of using protein spectra of microorganisms obtained by MALDI-ToF mass spectrometry as additional microbiological criteria in assessing the course of the infection process caused by Burkholderia cepacia complex among patients with cystic fibrosis. The analysis of protein profiles, which were obtained by using MALDI-ToF mass spectrometry (Bruker Daltonik GmbH, Germany), was performed by using flexAnalisis 3.0 software (Bruker Daltonik GmbH, Germany). Differences in protein profiles of Burkholderia spp. isolates were found depended on the stage of infection of a cystic fibrosis patient with prolonged colonisation of the lower respiratory tract. The protein profiles of Burkholderia spp. isolates that formed a heterogeneous population containing both NCV and SCV morphotypes were also studied. A regular dynamic monitoring and comparison of protein profiles of microbial strains can be useful in forecasting effort of the clinical course of the disease, as well as in assessing of risks of severe infectious complications development.

PMID:40655327 | PMC:PMC12246321 | DOI:10.1007/s12088-024-01366-8

bioRxiv [Preprint]. 2025 May 12:2025.05.07.652658. doi: 10.1101/2025.05.07.652658.

ABSTRACT

Staphylococcus aureus (SA), the most common cystic fibrosis (CF) lung pathogen, is uniquely capable of producing superantigen (SAg) exotoxins, which are the most potent activators of the immune system. Although the proinflammatory roles of SA-SAgs is well-established, their role in the immunopathogenesis of CF lung disease is unexplored. Herein, we demonstrate that 60-80% of pediatric and adult CF SA isolates carried at least one SA-SAg gene, with the former harboring potent SA-SAgs (Staphylococcal enterotoxin A and B) more frequently (30-60%). Biofilms of clinical SA isolates readily produced biologically active SA-SAgs in artificial sputum medium and purified SA-SAgs retained their bioactivity in human CF sputum in vitro. Repeated intratracheal challenge with purified SA-SAgs induced a robust pulmonary inflammatory response in CF mouse models (βENAC and CFGC transgenic mice) expressing HLA-DR3 in a dose-dependent manner, with the low dose favoring a type 2 eosinophilic lung inflammatory response, and a high dose eliciting a type 1 inflammatory response with neutrophilic lung inflammation and higher mortality. In vivo neutralization of IFN-γ also promoted SA-SAg-driven type 2 inflammation. Intratracheal infection with sub-lethal dose of a clinical SA isolate producing SEB, but not the SEB-deficient mutant isogenic SA strain, also elicited an eosinophilic inflammatory response.

PMID:40654878 | PMC:PMC12248047 | DOI:10.1101/2025.05.07.652658