Search Results (27)

Background/Objectives: In children, gastroesophageal reflux disease (GERD) may lead to epithelial barrier dysfunction and the release of thymic stromal lymphopoietin (TSLP), interleukin-25 (IL-25), interleukin-33 (IL-33) and periostin, known as alarmins. These cytokines are associated with type 2 inflammation and may contribute to respiratory and allergic conditions. The main purpose of this study is to evaluate serum concentrations of TSLP, IL-25, IL-33, and periostin in children with and without GERD and to assess their relationships with bronchial hyperresponsiveness (BHR) and sensitization to inhaled allergens. Methods: The study included 93 children aged 7–17 years. GERD was diagnosed based on 24-h esophageal pH impedance monitoring. Serum levels of TSLP, IL-25, IL-33, and periostin were measured using enzyme-linked immunosorbent assay (ELISA). It should be noted that the assay used does not distinguish between TSLP isoforms, which represents a limitation of the study. BHR was assessed via a methacholine challenge test, and allergen sensitization was determined using skin prick tests and allergen-specific immunoglobulin E (asIgE). Results: Serum TSLP levels were significantly higher in children with GERD compared to those without, whereas IL-25, IL-33 and periostin did not differ notably between groups. Periostin was associated with the degree of sensitization to inhalant allergens, but no significant links were found between cytokine levels and bronchial hyperresponsiveness. Conclusions: Significantly higher TSLP levels were noted in children with GERD than in those without. Hence, TSLP may have a potential role as a biomarker of epithelial immune activation in pediatric GERD. In addition, periostin was associated with sensitization to inhalant allergens, although it did not differentiate between children with and without GERD. Full article

Introduction: Asthma is a chronic airway inflammatory disease characterized by variable airflow limitation and intermittent symptoms. In well-controlled asthma, auscultation and spirometry often appear normal, making diagnosis challenging. Moreover, bronchial provocation tests carry a risk of inducing acute bronchoconstriction. This study aimed to develop a non-invasive, objective, and reproducible diagnostic method using machine learning-based lung sound analysis for the early detection of asthma, even during stable periods. Methods: We designed a machine learning algorithm to classify controlled asthma patients and healthy individuals using respiratory sounds recorded with a digital stethoscope. We enrolled 120 participants (60 asthmatic, 60 healthy). Controlled asthma was defined according to Global Initiative for Asthma (GINA) criteria and was supported by normal spirometry, no pathological auscultation findings, and no exacerbations in the past three months. A total of 3600 respiratory sound segments (each 3 s long) were obtained by dividing 90 s recordings from 120 participants (60 asthmatic, 60 healthy) into non-overlapping clips. The samples were analyzed using Mel-Frequency Cepstral Coefficients (MFCCs) and Tunable Q-Factor Wavelet Transform (TQWT). Significant features selected with ReliefF were used to train Quadratic Support Vector Machine (SVM) and Narrow Neural Network (NNN) models. Results: In 120 participants, pulmonary function test (PFT) results in the asthma group showed lower FEV1 (86.9 ± 5.7%) and FEV1/FVC ratios (86.1 ± 8.8%) compared to controls, but remained within normal ranges. Quadratic SVM achieved 99.86% accuracy, correctly classifying 99.44% of controls and 99.89% of asthma cases. Narrow Neural Network achieved 99.63% accuracy. Sensitivity, specificity, and F1-scores exceeded 99%. Conclusion: This machine learning-based algorithm provides accurate asthma diagnosis, even in patients with normal spirometry and clinical findings, offering a non-invasive and efficient diagnostic tool. Full article

Tuberculous meningoencephalitis is a rare manifestation of Mycobacterium tuberculosis (Mtb), with the most severe form and highest mortality. It can cause multiple complications, and treatment is difficult, as drugs cannot properly diffuse through the haemato-encephalitic barrier. We reported the case of a 17-year-old female patient who was admitted to the emergency room department with a fever for previous two weeks (up to 39 °C), dizziness, difficulty walking, and weight loss. Magnetic resonance imaging indicated possible meningoencephalitis, and a CT scan of the lungs visualised miliary infiltrates in both lungs. After repeated tests, Mtb DNA was found in the bronchial wash, cerebrospinal fluid, faeces, and urine via an Xpert/Rif Ultra test. Treatment was started with isoniazid, rifampicin, ethambutol, pyrazinamide, and corticosteroids as well. Although treatment was initiated within the first few days in the hospital, a reduction in glucocorticoid dosage worsened the patient’s neurological state, making treatment even more challenging. Prolonged use of glucocorticoids led to an improvement in the stage of the condition. Further, over time, the patient’s condition improved. Pulmonary infiltrations were not found after 2.5 months of starting therapy. Conclusions: Timely treatment is crucial for improving the prognosis of patients with miliary tuberculosis and tuberculous meningoencephalitis. Prompt recognition of symptoms and accurate diagnosis are essential to initiate effective treatment strategies. In this patient’s case, prolonged use of corticosteroids reduced neurologic complications, and ongoing treatment gradually improved the patient’s condition. Full article

Background and Objectives: Klebsiella pneumoniae is a major nosocomial pathogen with a remarkable ability to develop resistance to multiple antibiotics, posing significant treatment challenges. This study aims to evaluate the antimicrobial resistance patterns among multidrug-resistant (MDR) and non-MDR strains of K. pneumoniae isolated over a six-year period (2018–2023) at the Clinical Hospital of Infectious Diseases and Pulmonology “Dr. Victor Babes” in Timisoara, Romania. The objectives include categorizing isolates based on their antibiotic resistance profiles and identifying trends in resistance to key antibiotics to optimize treatment strategies and enhance infection control measures. Materials and Methods: A cross-sectional analysis was conducted on K. pneumoniae isolates obtained from various clinical samples between January 2018 and December 2023. Identification was performed using standard bacteriological procedures, and antimicrobial susceptibility testing was conducted using the Kirby–Bauer disk diffusion method in accordance with EUCAST guidelines. Isolates were classified as susceptible, resistant, MDR, extensively drug-resistant (XDR), or pandrug-resistant (PDR) based on ECDC definitions. Data were analyzed using GraphPad Prism 6, with chi-square tests and Cochran–Armitage trend tests applied where appropriate. Statistical significance was set at p < 0.05. Results: A total of 1,081 K. pneumoniae isolates were identified over the six-year period, increasing from 118 isolates in 2018 to 319 in 2023. The proportion of XDR and PDR strains showed a significant upward trend from 30.5% in 2018 to 57.4% in 2023 (p < 0.001). Specifically, XDR strains increased from 22.9% in 2018 to 39.8% in 2023, while PDR strains rose from 7.6% to 17.6%. Among monomicrobial infections in 2023, XDR and PDR strains accounted for 42.4% and 16.5%, respectively. Resistance to carbapenems also showed a significant increase; for instance, resistance to ertapenem rose from 35.6% in 2018 to 54.2% in 2023 (p < 0.001). Subgroup analysis revealed that isolates from bronchial aspirates had the highest rates of XDR and PDR strains in 2023, at 38.0% and 17.2%, respectively. Additionally, polymicrobial infections where both K. pneumoniae and co-infecting pathogens were XDR/PDR increased from 24.2% in 2018 to 46.6% in 2023 (p < 0.001). Conclusions: The study demonstrates a significant escalation in antimicrobial resistance among K. pneumoniae isolates over the six-year period, particularly in XDR and PDR strains. The rising trend of resistance to critical antibiotics like carbapenems underscores the urgent need for enhanced antimicrobial stewardship and infection control measures. Targeted interventions are essential to curb the spread of these resistant strains and to optimize therapeutic strategies. Full article

Anaphylaxis is a potentially life-threatening systemic allergic reaction that can result in fatal outcomes if not promptly and appropriately treated. The diagnosis of the cause of anaphylaxis during anesthesia can be challenging due to the complexity of the perioperative environment. Propofol-induced perioperative anaphylaxis is uncommon, occurring in perioperative anaphylactic shock cases. We present a case of perioperative anaphylactic shock in a patient with no known allergies who had been exposed to the same anesthetic agents, propofol, rocuronium, and remifentanil, three times previously without incident. Cardiac arrest occurred 50 min after induction, which showed pulseless electrical activity with decreasing saturation without bronchial spasm and skin erythema or edema. After prompt and appropriate management including cardiopulmonary resuscitation, the patient recovered without complications. The diagnosis was confirmed as propofol-induced anaphylactic shock by an elevated serum tryptase level, measured in a timely manner, and by skin tests (skin prick test and intradermal test), which revealed strong hypersensitivity to propofol. This case is notable for the cardiovascular collapse that occurred without respiratory symptoms or skin manifestations, as well as the delayed onset of anaphylaxis (>50 min). This case underscores the importance of vigilance for anaphylaxis, even with repeated exposure to previously well-tolerated drugs, as sensitization can lead to more severe reactions. It also highlights the potential for anaphylaxis to occur outside the acute phase and without typical clinical features. Full article

Exercise-induced bronchoconstriction (EIB) is a common clinical entity in people with asthma. EIB is characterized by postexercise airway obstruction that results in symptoms such as coughing, dyspnea, wheezing, chest tightness, and increased fatigue. The underlying mechanism of EIB is not completely understood. “Osmotic theory” and “thermal or vascular theory” have been proposed. Initial assessment must include a specific work-up to exclude alternative diagnoses like exercise-induced laryngeal obstruction (EILO), cardiac disease, or physical deconditioning. Detailed medical history and clinical examination must be followed by basal spirometry and exercise challenge test. The standardized treadmill running (TR) test, a controlled and standardized method to assess bronchial response to exercise, is the most adopted exercise challenge test for children aged at least 8 years. In the TR test, the goal is to reach the target heart rate in a short period and maintain it for at least 6 min. The test is then followed by spirometry at specific time points (5, 10, 15, and 30 min after exercise). In addition, bronchoprovocation tests like dry air hyperpnea (exercise and eucapnic voluntary hyperpnea) or osmotic aerosols (inhaled mannitol) can be considered when the diagnosis is uncertain. Treatment options include both pharmacological and behavioral approaches. Considering medications, the use of short-acting beta-agonists (SABA) just before exercise is the commonest option strategy, but daily inhaled corticosteroids (ICS) can also be considered, especially when EIB is not controlled with SABA only or when the patients practice physical activity very often. Among the behavioral approaches, warm-up before exercise, breathing through the nose or face mask, and avoiding polluted environments are all recommended strategies to reduce EIB risk. This review summarizes the latest evidence published over the last 10 years on the pathogenesis, diagnosis using spirometry and indirect bronchoprovocation tests, and treatment strategies, including SABA and ICS, of EIB. A specific focus has been placed on EIB management in young athletes, since this condition can not only prevent them from practicing regular physical activity but also competitive sports. Full article

Background: Early diagnosis of ventilator-associated pneumonia (VAP) remains a challenge due to subjective clinical criteria and the low discriminative power of diagnostic tests. We assessed whether rapid molecular diagnostics in combination with Clinically Pulmonary Index Score (CPIS) scoring, microbiological surveillance and biomarker measurements of PTX-3, SP-D, s-TREM, PTX-3, IL-1β and IL-8 in the blood or lung could improve the accuracy of VAP diagnosis and follow-up in critically ill children. Methods: A prospective pragmatic study in a Pediatric Intensive Care Unit (PICU) was conducted on ventilated critically ill children divided into two groups: high and low suspicion of VAP according to modified Clinically Pulmonary Index Score (mCPIS). Blood and bronchial samples were collected on days 1, 3, 6 and 12 after event onset. Rapid diagnostics were used for pathogen identification and ELISA for PTX-3, SP-D, s-TREM, IL-1β and IL-8 measurements. Results: Among 20 enrolled patients, 12 had a high suspicion (mCPIS > 6), and 8 had a low suspicion of VAP (mCPIS < 6); 65% were male; and 35% had chronic disease. IL-1β levels at day 1 correlated significantly with the number of mechanical ventilation days (r_s = 0.67, p < 0.001) and the PICU stay (r = 0.66; p < 0.002). No significant differences were found in the levels of the other biomarkers between the two groups. Mortality was recorded in two patients with high VAP suspicion. Conclusions: PTX-3, SP-D, s-TREM, IL-1β and IL-8 biomarkers could not discriminate patients with a high or low suspicion of VAP diagnosis. Full article

Bronchial provocation tests, such as the mannitol challenge, can be performed to identify and quantify the severity of bronchial hyperresponsiveness in asthmatic patients. Studies of the mannitol challenge as a monitoring tool in asthmatic children are limited. Our primary aim was to compare the bronchial hyperresponsiveness to mannitol in treatment-naive asthmatic children between baseline and three months after receiving the indicated asthma prophylaxis. Twenty-three asthmatic patients aged 4–16 years were analyzed in this prospective cohort study. All subjects underwent the mannitol challenge at baseline and after three months of treatment with budesonide ± formoterol. The difference in the provocative dose of mannitol to induce a 15% drop in FEV₁ (PD₁₅) between baseline and follow-up, as well as its association with the presence of exercise-induced or nocturnal asthma symptoms, were evaluated. The PD₁₅ value increased significantly post-treatment (228.5 mg [4.50–458.15]; p = 0.04). Independently of the evaluation time point, the PD₁₅ values were significantly lower in the presence of nocturnal asthma symptoms (490 mg [122–635] vs. 635 mg [635–635]; p = 0.03), whereas there was no association between the PD₁₅ value and the presence of exercise-induced asthma (p = 0.73). These results suggest that bronchial hyperresponsiveness to mannitol may be a potential monitoring tool in the pediatric asthmatic population, reflecting therapy response in children receiving prophylactic treatment. Full article

Pseudomonas aeruginosa is a major hospital-associated pathogen that can cause severe infections, most notably in patients with cystic fibrosis (CF) or those hospitalized in intensive care units. Given its remarkable ability to resist antibiotics, P. aeruginosa eradication has grown more challenging. Therefore, there is an urgent need to discover and develop new strategies that can counteract P. aeruginosa-resistant strains. Here, we evaluated the efficacy of poly-L-lysine (pLK) in combination with commonly used antibiotics as an alternative treatment option against P. aeruginosa. First, we demonstrated by scanning electron microscopy that pLK alters the integrity of the surface membrane of P. aeruginosa. We also showed using a fluorometry test that this results in an enhanced permeability of the bacteria membrane. Based on these data, we further evaluated the effect of the combinations of pLK with imipenem, ceftazidime, or aztreonam using the broth microdilution method in vitro. We found synergies in terms of bactericidal effects against either sensitive or resistant P. aeruginosa strains, with a reduction in bacterial growth (up to 5-log₁₀ compared to the control). Similarly, these synergistic and bactericidal effects were confirmed ex vivo using a 3D model of human primary bronchial epithelial cells maintained in an air–liquid interface. In conclusion, pLK could be an innovative antipseudomonal molecule, opening its application as an adjuvant antibiotherapy against drug-resistant P. aeruginosa strains. Full article

Quantification of oxidative stress is a challenging task that can help in monitoring chronic inflammatory respiratory airway diseases. Different studies can be found in the literature regarding the development of electrochemical sensors for H₂O₂ in cell culture medium to quantify oxidative stress. However, there are very limited data regarding the impact of the cell culture medium on the electrochemical quantification of H₂O₂. In this work, we studied the effect of different media (RPMI, MEM, DMEM, Ham’s F12 and BEGM/DMEM) on the electrochemical quantification of H₂O₂. The used electrode is based on reduced graphene oxide (rGO) and gold nanoparticles (AuNPs) and was obtained by co-electrodeposition. To reduce the electrode fouling by the medium, the effect of dilution was investigated using diluted (50% v/v in PBS) and undiluted media. With the same aim, two electrochemical techniques were employed, chronoamperometry (CH) and linear scan voltammetry (LSV). The influence of different interfering species and the effect of the operating temperature of 37 °C were also studied in order to simulate the operation of the sensor in the culture plate. The LSV technique made the sensor adaptable to undiluted media because the test time is short, compared with the CH technique, reducing the electrode fouling. The long-term stability of the sensors was also evaluated by testing different storage conditions. By storing the electrode at 4 °C, the sensor performance was not reduced for up to 21 days. The sensors were validated measuring H₂O₂ released by two different human bronchial epithelial cell lines (A549, 16HBE) and human primary bronchial epithelial cells (PBEC) grown in RPMI, MEM and BEGM/DMEM media. To confirm the results obtained with the sensor, the release of reactive oxygen species was also evaluated with a standard flow cytometry technique. The results obtained with the two techniques were very similar. Thus, the LSV technique permits using the proposed sensor for an effective oxidative stress quantification in different culture media and without dilution. Full article

Introduction: Asthma is characterized by persistent inflammation, airway hypersensitivity and remodelling. Bone Morphogenetic Proteins belong to the Transforming Growth Factor Superfamily and have a similar signalling transduction pathway and common co-mediating protein. However, the BMPs role in the remodelling remains unclear; they appear to be involved in the airway inflammation and fibrogenesis process. Material and Methods: 60 patients with asthma and 48 healthy volunteers were recruited for the study. Blood samples were collected before, 1 h, 24 and 48 h after the allergen or the methacholine challenge test. Evaluation of BMP-4 and BMP-7 serum concentration and expression was performed using ELISA and real time PCR methods, respectively. Results: Statistically significant differences in BMP-7 concentration between healthy controls and asthmatics before the chal-lenge were noted. We found two statistically significant correlations: between the basal BMP-4 concentration and the FEV1(L) raw value and FEV1/FVC(%) index. We did not observe significant changes in the gene expression of BMP-4 and BMP-7 in different time points. Conclusions: Observed differences in BMP-7 concentration between asthmatic and healthy groups and correlations between BMP-4 concentration and some lung function test values may indicate the role of the BMPs in the etiopathogenesis of asthma. The unique characteristic of our study is the evaluation of BMPs serum levels, not in the bronchial epithelium. Full article

Lung cancer has the highest mortality rate worldwide and is often diagnosed at late stages, requiring genotoxic chemotherapy with significant side effects. Cancer prevention has become a major focus, including the use of dietary and supplemental antioxidants. Thus, we investigated the ability of an antioxidant formulation (AOX1) to reduce DNA damage in human bronchial epithelial cells (BEAS-2B) with and without the combination of apple peel flavonoid fraction (AF4), or its major constituent quercetin (Q), or Q-3-O-d-glucoside (Q3G) in vitro. To model smoke-related genotoxicity, we used cigarette-smoke hydrocarbon 4-[(acetoxymethyl)nitrosamino]-1-(3-pyridyl)-1-butanone (NNKOAc) as well as methotrexate (MTX) to induce DNA damage in BEAS-2B cells. DNA fragmentation, γ-H2AX immunofluorescence, and comet assays were used as indicators of DNA damage. Pre-exposure to AOX1 alone or in combination with AF4, Q, or Q3G before challenging with NNKOAc and MTX significantly reduced intracellular reactive oxygen species (ROS) levels and DNA damage in BEAS-2B cells. Although NNKOAc-induced DNA damage activated ATM-Rad3-related (ATR) and Chk1 kinase in BEAS-2B cells, pre-exposure of the cells with tested antioxidants prior to carcinogen challenge significantly reduced their activation and levels of γ-H2AX (p ≤ 0.05). Therefore, AOX1 alone or combined with flavonoids holds promise as a chemoprotectant by reducing ROS and DNA damage to attenuate activation of ATR kinase following carcinogen exposure. Full article

The management of complex airway disorders is challenging, as the airway stent placement usually results in several complications. Tissue reaction to the foreign body, poor mechanical properties and inadequate fit of the stent in the airway are some of the reported problems. For this reason, the design of customized biomedical devices to improve the accuracy of the clinical results has recently gained interest. The aim of the present study is to introduce a parametric tool for the design of a new tracheo-bronchial stent that could be capable of improving some of the performances of the commercial devices. The proposed methodology is based on the computer aided design software and on the finite element modeling. The computational results are validated by a parallel experimental work that includes the production of selected stent configurations using the 3D printing technology and their compressive test. Full article

Work-related asthma (WRA) is a very frequent condition in the occupational setting, and refers either to asthma induced (occupational asthma, OA) or worsened (work-exacerbated asthma, WEA) by exposure to allergens (or other sensitizing agents) or to irritant agents at work. Diagnosis of WRA is frequently missed and should take into account clinical features and objective evaluation of lung function. The aim of this overview on pulmonary function testing in the field of WRA is to summarize the different available tests that should be considered in order to accurately diagnose WRA. When WRA is suspected, initial assessment should be carried out with spirometry and bronchodilator responsiveness testing coupled with first-step bronchial provocation testing to assess non-specific bronchial hyper-responsiveness (NSBHR). Further investigations should then refer to specialists with specific functional respiratory tests aiming to consolidate WRA diagnosis and helping to differentiate OA from WEA. Serial peak expiratory flow (PEF) with calculation of the occupation asthma system (OASYS) score as well as serial NSBHR challenge during the working period compared to the off work period are highly informative in the management of WRA. Finally, specific inhalation challenge (SIC) is considered as the reference standard and represents the best way to confirm the specific cause of WRA. Overall, clinicians should be aware that all pulmonary function tests should be standardized in accordance with current guidelines. Full article

Occupational asthma (OA) represents one of the major public health problems due to its high prevalence, important social and economic burden. The aim of this review is to summarize current data about clinical phenotypes, biomarkers, diagnosis and management of OA, a subtype of work-related asthma. Most studies have identified two phenotypes of OA. One is sensitizer-induced asthma, occuring after a latency period and caused by hypersensitivity to high- or low-molecular weight agents. The other is irritant-induced asthma, which can occur after one or more exposures to high concentrations of irritants without latency period. More than 400 agents causing OA have been identified and its list is growing fast. The best diagnostic approach for OA is a combination of clinical history and objective tests. An important tool is a specific inhalation challenge. Additional tests include assessments of bronchial hyperresponsiveness to methacholine/histamine in patients without airflow limitations, monitoring peak expiratory flow at- and off-work, sputum eosinophil count, exhaled nitric oxide measurement, skin prick tests with occupational allergens and serum specific IgE. Treatment of OA implies avoidance of exposure, pharmacotherapy and education. OA is a heterogeneous disease. Mechanisms of its different phenotypes, their diagnosis, role of new biomarkers and treatment require further investigation. Full article