Natural antibodies targeting LPS in pleural effusions of various etiologies.

Respiratory infection, cancer, and heart failure can cause abnormal accumulation of fluid in the pleural cavity. The immune responses within the cavity are orchestrated by leucocytes that reside in the serosal-associated lymphoid tissue. Natural antibodies (NAbs) are abundant in the serum (S) having a major role in systemic and mucosal immunity; however, their occurrence in pleural fluid (PF) remains an open question. Our aim herein was to detect and measure the levels of NAbs (IgM, IgG, IgA) targeting lipopolysaccharides (LPS) in both the pleural fluid and the serum of 78 patients with pleural effusions (PEs) of various etiologies. The values of anti-LPS NAb activity were extracted through a normalization step regarding the total IgM, IgG, and IgA levels, all determined by in-house ELISA. In addition, the ratios of PF/S values were analyzed further with other critical biochemical parameters from pleural fluids. Anti-LPS NAbs of all Ig classes were detected in most of the samples, while a significant increase of anti-LPS activity was observed in infectious and noninfectious compared with malignant PEs. Multivariate linear regression confirmed a negative correlation of IgM and IgA anti-LPS PF/S ratio with malignancy. Moreover, anti-LPS NAbs PF/S measurements led to increased positive and negative predictive power in ROC curves generated for the discrimination between benign and malignant PEs. Our results highlight the role of anti-LPS NAbs in the pleural cavity and demonstrate the potential translational impact that should be further explored.NEW & NOTEWORTHY Here we describe the detection and quantification of natural antibodies (NAbs) in the human pleural cavity. We show for the first time that IgM, IgG, and IgA anti-LPS natural antibodies are detected and measured in pleural effusions of infectious, noninfectious, and malignant etiologies and provide clinical correlates to demonstrate the translational impact of our findings.

Pioglitazone and asthma: a review of current evidence.

OBJECTIVE: This review aims to present existing evidence on the impact of pioglitazone, a thiazolidinedione class anti-diabetic drug, on asthma control and lung function, providing a comprehensive understanding of its potential as a treatment for asthma. DATA SOURCES: The review draws upon data from preclinical animal studies and clinical trials investigating the effects of pioglitazone on asthma, focusing on its role in reducing airway inflammation, hyperreactivity, and remodeling, and its impact on pulmonary function. STUDY SELECTIONS: Relevant studies were selected based on their examination of pioglitazone's therapeutic effects in asthma, including both animal models and clinical trials involving human asthma patients. RESULTS: Animal studies have suggested that pioglitazone could alleviate inflammation, airway hyperreactivity, and airway remodeling, thereby improving pulmonary function in asthma. However, clinical trials have not demonstrated significant therapeutic benefits, with minimal improvements observed in asthma control and lung function, and the presence of notable side effects. CONCLUSION: Despite promising preclinical data, the efficacy of pioglitazone in treating human asthma remains unproven, with safety concerns and limited clinical benefits observed in trials. Further research is needed to assess the safety and effectiveness of pioglitazone in asthma treatment and to explore its impact on other inflammatory mechanisms.

Psychopathology in asthma patients: analysis of demographic and clinical associations.

BACKGROUND: Asthma, a chronic inflammatory condition affecting the airways, significantly impacts both respiratory function and quality of life. Recent studies have highlighted the psychological dimensions of chronic diseases like asthma. Despite growing evidence linking asthma with various psychopathological conditions, comprehensive data remains scarce. AIM: This study aims to explore the psychopathological status of asthma patients and identify demographic and clinical factors associated with higher levels of psychopathological symptoms. METHODS: Data were collected from 42 asthma patients attending the Asthma Outpatient Clinic at the University of Thessaly, Greece. Participants completed a detailed questionnaire on demographics and clinical parameters, along with the Symptoms Checklist-90 (SCL-90) to assess psychological symptoms. RESULTS: The sample included patients with varying asthma severity: 38% with severe asthma and 62% with mild-moderate asthma, with a mean age of 60.8 ± 15.6 years. Results indicated that 23.8% exhibited somatization symptoms, 23.8% showed compulsive behaviors, 33.8% reported interpersonal sensitivity, and 38.1% experienced depression and 38.1% aggressiveness. Additionally, 45.2% struggled with fearful anxiety, 4.8% showed paranoid ideation, and 11.9% had traits of psychoticism. Women had significantly higher psychopathology scores than men. Factors such as longer disease duration, uncontrolled asthma, severe asthma, and comorbid conditions like atopy and gastroesophageal reflux disease (GERD) were linked to higher psychopathological scores. CONCLUSIONS: The study found a high prevalence of psychopathological symptoms among asthma patients. Female gender, prolonged asthma duration, persistent symptoms, comorbid diseases (GERD, atopy) and greater disease severity were significantly associated with higher psychopathology, underscoring the need for integrated mental health care in asthma management.

Effects of pharmacological primary cilium disturbance in the context of in vitro 2D and 3D malignant pleura mesothelioma.

INTRODUCTION: Primary cilium (PC) is a single non-motile antenna-like organelle composed of a microtubule core axon originating from the mother centriole of the centrosome. The PC is universal in all mammalian cells and protrudes to the extracellular environment receiving mechanochemical cues that it transmits in the cell. AIM: To investigate the role of PC in mesothelial malignancy in the context of two-dimensional (2D) and three-dimensional (3D) phenotypes. MATERIALS AND METHODS: The effect of pharmacological deciliation [using ammonium sulphate (AS) or chloral hydrate (CH)] and PC elongation [using lithium chloride (LC)] on cell viability, adhesion, and migration (2D cultures) as well as in mesothelial sphere formation, spheroid invasion and collagen gel contraction (3D cultures) was investigated in benign mesothelial MeT-5A cells and in malignant pleural mesothelioma (MPM) cell lines, M14K (epithelioid) and MSTO (biphasic), and primary malignant pleural mesothelioma cells (pMPM). RESULTS: Pharmacological deciliation or elongation of the PC significantly affected cell viability, adhesion, migration, spheroid formation, spheroid invasion and collagen gel contraction in MeT-5A, M14K, MSTO cell lines and in pMPM cells compared to controls (no drug treatment). CONCLUSIONS: Our findings indicate a pivotal role of the PC in functional phenotypes of benign mesothelial cells and MPM cells.

Work place and prevalence of COVID-19 in a rural population in Greece.

INTRODUCTION: Coronavirus disease-19 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has become a global pandemic, giving rise to a serious global health threat. Many countries including Greece have seen a two-wave pattern of reported cases, with a first wave in spring and a second in autumn of 2020. METHODS: A cross-sectional seroprevalence study was designed to measure the prevalence of IgG antibodies with a quantitative SARS-CoV-2 IgG lab-based serology test, chemiluminescent microparticle immunoassay, against novel coronavirus in rural areas in Greece after the second pandemic wave. The study was conducted on 29 January 2021 in a rural semi-closed area, the municipality of Deskati, prefecture of western Macedonia in Greece after the second pandemic wave. RESULTS: Sixty-nine participants were included in this study. The present study demonstrated a high prevalence of COVID-19 infection (31 of 69 total participants; 45%) and those who were working in the public sector were at higher risk of COVID-19 infection in comparison to their counterparts in private sector (p=0.05364), (relative risk 2.64; 95% confidence interval 1.001-7.086). CONCLUSION: The study presents data showing a high prevalence of herd immunity for COVID-19 in a semi-closed area in Greece. These findings might help to understand the characteristics of this second wave, the behaviour and danger of SARS-CoV-2 in rural areas in Greece and Europe generally.

HΜGB1/sRAGE levels differ significantly between transudates and exudates.

High mobility group box 1(HMGB1) protein operates as an alarmin with multiple roles in immunity and cell homeostasis. It is highly expressed in epithelial barrier sites and acts via the binding to the receptor for advanced glycation end products (RAGE). Production of HMGB1 and soluble RAGE (sRAGE), a decoy receptor for HMGB1, has been implicated in several pulmonary diseases, but both have been scarcely investigated in pleural diseases. The aim of this study was to determine the levels of HMGB1 and sRAGE in transudative, malignant and parapneumonic pleural effusions (PEs) and to investigate the effect of low and high HMGB1 pleural fluid levels on MeT-5A cell adhesion, migration and spheroid formation, in each group. HMGB1 and sRAGE levels were significantly lower and higher in transudative PEs compared to malignant and parapneumonic PEs, respectively. Patients above 65 years of age had significantly lower HMGB1 and higher sRAGE levels compared to patients below 65 years old. Furthermore, incubation of MeT-5A cells with malignant or parapneumonic PEs bearing low or high levels of HMGB1 yielded significant differential effects on MeT-5A cell adhesion, migration and spheroid formation. In all types of effusions, high HMGB1 levels correlated with more adherence compared to low HMGB1 levels. In transudative and malignant PEs high HMGB1 levels correlated with decreased migration of MeT-5A cells while in parapneumonic ones the effect was the opposite. Only samples from parapneumonic PEs high in HMGB1 achieved uniform spheroid formation. These results reveal a clinical context-dependent effect of the HMGB1/sRAGE axis in PEs.

Clinical profile of severe asthmatics receiving biological treatment: see behind the scenes.

The clinical phenotyping of severe asthma could improve treatment outcomes, quality of life, morbidity, and mortality of asthma patients. The growing availability of rich clinical data could provide opportunities to address a broad range of real-world questions regarding clinical phenotyping and effective therapeutic approaches of severe asthma. Accordingly, in this Letter to the Editor, we provide data relevant to constructing the clinical profile of a Greek severe asthma patient cohort who receive biological treatment.

Serum vitamin D levels in patients with obstructive sleep apnea syndrome and level changes after continuous positive airway pressure therapy.

PURPOSE: Patients with obstructive sleep apnea syndrome (OSAS) have been found to exhibit lower serum vitamin D levels, even when the control groups are matched for confounding conditions. However, contradictory studies are also present. This study aimed to compare serum 25-hydroxyvitamin D (25(OH)D) levels between adult patients with OSAS and non-apneic controls and to evaluate the changes in 25(OH)D levels after 3 and 12 months of continuous positive airway pressure (CPAP) therapy. METHODS: The study was comprised of 30 patients with OSAS and 30 controls. Serum 25(OH)D levels were determined at baseline and after 3 and 12 months of CPAP therapy in all patients with OSAS. For analysis, patients with OSAS were divided into subgroups by adherence, with adherence defined as CPAP usage for > 4 h per night on at least 70% of nights. RESULTS: The 25(OH)D levels were not significantly different between OSAS and control groups at baseline. 25(OH)D levels did not change after 3 and 12 months of CPAP therapy. Patients who were CPAP-adherent showed less reduction in 25(OH)D levels compared with non-adherent ones (21.18 ± 9.3 vs. 12.13 ± 3.8 ng/mL, p = 0.022) after 1 year. The 25(OH)D levels were significantly correlated with higher daily CPAP usage at 3 and 12 months. Mean daily CPAP usage was a significant predictor of serum 25(OH)D levels at 12 months. CONCLUSIONS: Patients with OSAS who demonstrated good CPAP adherence showed significantly higher 25(OH)D levels after 1 year compared with those not adequately using CPAP. Long-term good CPAP adherence and highly daily CPAP usage positively affected 25(OH)D levels in patients with OSAS.

Calprotectin in Lung Diseases.

Calprotectin (CLP) is a heterodimer formed by two S-100 calcium-binding cytosolic proteins, S100A8 and S100A9. It is a multifunctional protein expressed mainly by neutrophils and released extracellularly by activated or damaged cells mediating a broad range of physiological and pathological responses. It has been more than 20 years since the implication of S100A8/A9 in the inflammatory process was shown; however, the evaluation of its role in the pathogenesis of respiratory diseases or its usefulness as a biomarker for the appropriate diagnosis and prognosis of lung diseases have only gained attention in recent years. This review aimed to provide current knowledge regarding the potential role of CLP in the pathophysiology of lung diseases and describe how this knowledge is, up until now, translated into daily clinical practice. CLP is involved in numerous cellular processes in lung health and disease. In addition to its anti-microbial functions, CLP also serves as a molecule with pro- and anti-tumor properties related to cell survival and growth, angiogenesis, DNA damage response, and the remodeling of the extracellular matrix. The findings of this review potentially introduce CLP in daily clinical practice within the spectrum of respiratory diseases.

Exhaled nitric oxide as a marker of chlorine exposure in young asthmatic swimmers.

BACKGROUND: Swimming is recommended for people with asthma. However, the inevitable exposure to chlorine and its disinfectant byproducts in indoor swimming pools could be responsible for bronchial inflammation and asthma development. Fractional exhaled nitric oxide (FeNO) is a noninvasive marker of airway inflammation that predicts asthma exacerbations. OBJECTIVES: To evaluate pretraining and posttraining FeNO levels in young swimmers with asthma attending an indoor chlorinated pool compared with a set of healthy swimmers and to examine the potential risk of exposure to chlorine as a factor associated with bronchial inflammation. METHODS: A total of 146 children (8-18 years old) constantly attending an indoor chlorinated swimming pool were enrolled. Spirometry and FeNO measurements were performed 30 minutes after their arrival at the pool and immediately after exercise. Pre-exercise and postexercise spirometric and FeNO levels were assessed in a random subgroup of 14 swimmers (10 with asthma and 4 without) who performed cardiopulmonary exercise testing. RESULTS: Asthma was detected in 23 swimmers. In swimmers with asthma, preswimming FeNO values were significantly elevated compared with swimmers without asthma and their FeNO values measured before cardiopulmonary exercise testing. Postexercise FeNO values were significantly decreased by approximately one-third in healthy children and children with asthma in all sporting backgrounds. However, postswimming FeNO values remained significantly higher in swimmers with asthma compared with those without asthma. Forced expiratory volume in 1 second (FEV1), forced vital capacity (FVC), and FEV1/FVC ratio values showed no significant difference before and after 2 types of activity. CONCLUSION: Elevated FeNO levels before and after swimming were recorded in swimmers with asthma not observed in a different exercise field. The presence of chlorine in the indoor swimming pool seems to explain this finding.

Repeatability of pleural adenosine deaminase measurements in diagnostic evaluation of pleural effusions.

BACKGROUND: A follow-up thoracentesis is proposed in suspected atypical tuberculosis cases. The study aimed to define the variability of pleural ADA values across repeated thoracenteses in different types of pleural effusions (PEs) and to evaluate whether ADA variance, in regard to the cutoff value of 40 U/L, affected final diagnosis. METHODS: A total of 131 patients with PEs of various etiologies underwent three repeated thoracenteses. ADA values were subsequently estimated. RESULTS: 82% and 55% of patients had greater than 10% and 20% deviation from the highest ADA value, respectively. From those patients who had a variance of 20%, 36% had only increasing ADA values, while 19% had only decreasing values. Considering the cutoff value of 40 U/L, only in two cases, ADA decreased below this threshold, which concerned a man with tuberculous pleurisy and a woman with lymphoma both in the course of treatment. Furthermore, only in two cases with rising values, ADA finally exceeded the cutoff limit, which concerned a man with rheumatoid pleurisy and a man with tuberculous pleurisy. Surprisingly, malignant PEs (MPEs) showed a higher percentage of increasing values compared to all other exudates that did not, however, exceed the threshold. CONCLUSION: The determination of pleural ADA levels is a reproducible method for rapid tuberculosis diagnosis. The detected measurement deviations do not appear to affect final diagnosis. In specific situations, repeated ADA measurements may be valuable in directing further diagnostic evaluation. More investigation is needed to elucidate the possible prognostic significance of the increasing trend in ADA values in MPEs.

Takotsubo cardiomyopathy: A known unknown foe of asthma.

INTRODUCTION: Patients with uncontrolled asthma are at a greater risk of asthma attacks requiring emergency room visits or hospital admissions. Takotsubo cardiomyopathy is potentially a significant complication in a course of status asthmaticus. CASE STUDY: We describe a 43-year-old female patient who presented with status asthmaticus that was further complicated with takotsubo cardiomyopathy. RESULTS: Recognizing apical ballooning syndrome is challenging in patients with a history of respiratory disease because the symptoms of the last entity may complicate the diagnostic approach. It is difficult to distinguish clinically apical ballooning syndrome from the acute airway exacerbation itself. Both asthma and takotsubo cardiomyopathy share the same clinical presentation with dyspnea and chest tightness. In our patient, the electrocardiographic abnormalities, the rapidly reversible distinctive characteristics of echocardiography, and the modest elevation of serum cardiac biomarkers levels, in combination with the presence of a stress trigger (severe asthma attack), strongly supported the diagnosis of broken heart syndrome. CONCLUSIONS: Clinicians should re-evaluate asthma management and be aware of the complications associated with asthma attacks such as stress-induced cardiomyopathy.

Leptospirosis Incidence Post-Flooding Following Storm Daniel: The First Case Series in Greece.

The present study investigates the public health impact of flooding on leptospirosis incidence after Storm Daniel in Thessaly, Greece, in September 2023. A notable increase in cases was observed, with seven cases of female patients and a mean age of 40.2 years, indicating a significant risk among working-age adults. From the end of September to the beginning of November 2023, a total of 35 patients from flood-prone areas presented to the Emergency Department of the Tertiary University Hospital of Larissa. Diagnosis of leptospirosis was established by meeting the criteria suggested by the national public health organisation (EODY)-compatible clinical course, epidemiological exposure, molecular and serologic confirmation by the detection of immunoglobulin M antibodies to leptospira spp. using a commercially available enzyme-linked immunosorbent assay and real-time quantitative PCR for the molecular detection of leptospira. The larger part (84.6%) of leptospirosis cases were associated with contact with floodwater. The majority of these patients (71.4%) were from the prefecture of Larissa, followed by 14.3% from the prefecture of Karditsa, 8.6% from the prefecture of Trikala, and 5.7% from the prefecture of Magnesia. Occupational exposure and urbanisation were key risk factors. The most prevalent clinical feature was rash (69.2%), followed by fever (61.5%) and myalgia (30.7%). The findings emphasise the need for robust public health strategies, improved sanitation, rodent control, and protective measures for sanitation workers. The data highlight the broader implications of climate change on public health and the necessity for ongoing surveillance and community education to mitigate future outbreaks.

Assessing the Impact of Spatial and Temporal Variability in Fine Particulate Matter Pollution on Respiratory Health Outcomes in Asthma and COPD Patients.

Ambient air pollution's health impacts are well documented, yet the domestic environment remains underexplored. We aimed to compare indoor versus outdoor (I/O) air quality and estimate the association between indoor/ambient fine particulate matter (PM2.5) exposure and lung function in asthma and chronic obstructive pulmonary disease (COPD) patients. The study involved 24 h monitoring of PM2.5 levels indoors and outdoors, daily peak expiratory flow (PEF), and biweekly symptoms collection from five patients with asthma and COPD (average age of 50 years, 40% male) over a whole year. Data analysis was performed with linear mixed effect models for PEF and generalized estimating equations (GEE) for exacerbations. More than 5 million PM2.5 exposure and meteorological data were collected, demonstrating significant I/O PM2.5 ratio variability with an average ratio of 2.20 (±2.10). Identified indoor PM2.5 sources included tobacco use, open fireplaces, and cooking, resulting in average indoor PM2.5 concentrations of 63.89 µg/m3 (±68.41), significantly exceeding revised World Health Organization (WHO) guidelines. Analysis indicated a correlation between ambient PM2.5 levels and decreased PEF over 0-to-3-day lag, with autumn indoor exposure significantly impacting PEF and wheezing. The study underscores the need to incorporate domestic air quality into public health research and policy-making. A personalized approach is required depending on the living conditions, taking into account the exposure to particulate pollution.

Assessment of Knowledge, Attitudes, and Vaccination Practices Regarding the New RSV Vaccine among Health Professionals in Greece.

The introduction of a new vaccine into immunization programs represents a significant advancement in the global effort to combat vaccine-preventable diseases. Data from the World Health Organization support that immunization prevents between 2 and 3 million deaths each year across various diseases, underscoring its pivotal role in global health. The present study aims to assess the knowledge, attitudes, and anticipated vaccination practices among health professionals in Central Greece in response to the potential introduction of new Respiratory Syncytial Virus (RSV) vaccination guidelines by the National Vaccines Committee. Among the 450 health professionals solicited for the study, 219 provided responses, yielding a response rate of approximately 55%. A substantial majority (70.3%) accurately identified the vaccine's current availability, and 62.1% were aware of the current recommendation for RSV vaccination in pregnant women. In response to whether health professionals support the inclusion of an RSV vaccine in the national vaccination program if it becomes commercially available and is recommended by the Greek National Immunization Program, general practitioners showed the most support, with an average score of 4.86 (95% CI, 4.69-5.00), followed by pediatricians at 4.76 (95% CI, 4.63-4.89), pulmonologists at 4.68 (95% CI, 4.36-5.00), and obstetricians at 4.33 (95% CI, 3.95-4.71). Concerning general opinions on vaccinations, a high level of agreement was noted among the majority of health professionals, excluding nurses. Pharmacists recorded the highest agreement, with a perfect score of 5 (CI, 5.00-5.00), followed closely by pediatricians at 4.99 (CI, 4.97-5.00), GPs at 4.95 (CI, 4.85-5.00), pulmonologists at 4.93 (CI, 4.83-5.00), obstetricians at 4.74 (CI, 4.42-5.00), and nurses at 3.80 (CI, 3.06-4.54). A tailored approach to education is needed to ensure that healthcare professionals can communicate more effectively about RSV risks and vaccination benefits, fostering a proactive stance towards disease prevention and patient care. In essence, our study underscores the importance of knowledge in shaping a compassionate and responsive healthcare environment, ready to meet the challenges of RSV head-on.

Detection and Characterization of Extracellular Vesicles in Sputum Samples of COPD Patients.

BACKGROUND: Only one study has reported the presence of extracellular vesicles (EVs) in COPD patients' sputum. Thus, we aimed to isolate and characterize EVs from COPD and healthy individuals' sputum. METHODS: A total of 20 spontaneous sputum samples from COPD patients (m/f: 19/1) and induced sputum samples from healthy controls (m/f: 8/2) were used for EV isolation. The sputum supernatants were resuspended in PBS, precleared by centrifugation at 800× g for 10 min at 4 °C, and passed through a 0.22 µm filter (Millipore, Burlington, MA, USA). EVs were isolated by a standard membrane affinity spin column method (exoEasy maxi kit, Qiagen, Hilden, Germany). The EVs were then characterized by assessing their morphology and size using Transmission Electron Microscopy (TEM) and determining the CD9 and CD81 EV-markers with Western blot analysis. RESULTS: The EVs had a spherical shape and their mean diameter in the COPD patients was significantly greater than in the controls. Enrichment of the EV markers, CD9 and CD81, were detected in both the healthy and COPD individuals. Total EV-associated protein was significantly increased in the COPD patients compared to the controls. ROC analysis showed that total EV-associated protein in the sputum could be used to differentiate between the controls and COPD patients, with a sensitivity of 80% and a specificity of 70% at a cut-off point of 55.59 µg/mL (AUC = 0.8150). CONCLUSIONS: EVs were detectable in both the COPD and healthy individuals' sputum. The ratio of EVs in the 150-200 nm range was twice as high in the COPD patients than in the controls. The COPD patients' sputum contained increased total EV-associated protein as compared to controls, highlighting their value as a new source of specific exoproteins.