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BACKGROUND: Respiratory infections pose a major public health threat. The predominant viruses causing viral respiratory infections are influenza A and B (Flu-A, Flu-B), coronaviruses, respiratory syncytial virus (RSV), and adenovirus. This study aims to investigate the proportion of these cases via rapid antigen tests and assess seasonal patterns. METHODS: Clinical samples were collected from symptomatic adults presenting to the Emergency and Respiratory Medicine Departments of the University Hospital of Larissa (UHL), Greece from 16 October 2023 to 31 March 2024. Nasal specimens were antigen-tested for Flu-A/B, SARS-CoV-2, RSV, and adenovirus. RESULTS: The total sample of specimens collected was 1434, of which 739 (51.5%) were female and 695 were male (48.5%). The mean age of participants was 57 ± 5.5 years. Among the positive results, we recorded a proportion of 40.18% and 11.40% for influenza A and B, respectively, followed by 35.79% for SARS-CoV-2, 10.70% for RSV, and 1.93% for adenovirus. CONCLUSIONS: In Greece, surveillance systems in infection control are underutilized. Rapid tests via multiple antigens can quickly identify viral infections, making them a valuable tool with financial benefits for health systems. Early detection of respiratory infections helps allocate resources efficiently, ensures adequate staff and facilities are available, and improves patient care through refined clinical management.

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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.

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BACKGROUND & METHODS: This paper describes a pilot application of the Epidemic Volatility Index (EVI) to data from the pulmonary clinic of the University Hospital of Thessaly, Greece, for monitoring respiratory infections, COVID-19, and flu cases. EVI, a simple and easily implemented early warning method based on the volatility of newly reported cases, exhibited consistent and stable performance in detecting new waves of epidemics. The study highlights the importance of implementing early warning tools to address the effects of epidemics, including containment of outbreaks, timely intervention strategies, and resource allocation within real-world clinical settings as part of a broader public health strategy. RESULTS: The results presented in the figures demonstrate the association between successive early warnings and the onset of new waves, providing valuable insights for proactive decision-making. A web-based application enabling real-time monitoring and informed decision-making by healthcare professionals, public health officials, and policymakers was developed. CONCLUSIONS: This study emphasizes the significant role of early warning methods in managing epidemics and safeguarding public health. Future research may explore extensions and combinations of multiple warning systems for optimal outbreak interventions and application of the methods in the context of personalized medicine.

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We investigated the antibody kinetics after vaccination against COVID-19 in healthcare workers of a Greek tertiary hospital. Eight hundred and three subjects were included, of whom 758 (94.4%) received the BNT162b2 vaccine (Pfizer-BioNTech), eight (1%) mRNA-1273 (Moderna), 14 (1.7%) ChAdOx1 (Oxford-AstraZeneca) and 23 (2.9%) Ad26.COV2.S (Janssen). Before the second dose, at 2, 6 and 9 months after the second dose and at 2 and 6 months after the third dose, anti-spike IgG were quantified by the chemiluminescence microparticle immunoassay method. One hundred subjects were infected before vaccination (group A), 335 were infected after receiving at least one vaccine dose (group B), while 368 had never been infected (group C). Group A presented a greater number of hospitalizations and reinfections compared to group B (p < 0.05). By multivariate analysis, younger age was associated with an increased risk of reinfection (odds ratio: 0.956, p = 0.004). All subjects showed the highest antibody titers at 2 months after the second and third dose. Group A showed higher antibody titers pre-second dose, which remained elevated 6 months post-second dose compared to groups B and C (p < 0.05). Pre-vaccine infection leads to rapid development of high antibody titer and a slower decline. Vaccination is associated with fewer hospitalizations and fewer reinfections.

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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.

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BACKGROUND/AIM: The relationship between the kinetics of antibody responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and the severity of Coronavirus Disease 2019 (COVID-19) is poorly understood. The aim of the present study was to investigate whether serum SARS-CoV-2 antibody kinetics serve as an early predictor of clinical deterioration or recovery in hospitalized patients with COVID-19. PATIENTS AND METHODS: In this prospective observational study, 102 consecutive patients (median age: 60 years, 58% males) with symptomatic COVID-19 infection diagnosed by real-time polymerase chain reaction assay, hospitalized in two tertiary hospitals, were included. Rapid test for qualitative detection of immunoglobulin M (IgM) and immunoglobulin G (IgG) SARS-CoV-2 antibodies was performed at pre-defined time intervals during hospitalization (days: 0, 3, 7, 10, 14, 21 and 28). RESULTS: During a 3-month follow-up period after COVID-19 disease onset, a total of 87 patients were discharged, 12 patients were intubated and entered the Intensive Care Unit, and three patients died. The median time for seroconversion was 10 days for IgM and 12 days for IgG post onset of symptoms. Univariate logistic regression analysis found no associations between IgM or IgG positivity and clinical outcomes or complications during hospitalization for COVID-19 infection. Diabetes and dyslipidemia were the only clinical risk factors predictive of COVID-19-related complications during hospitalization. CONCLUSION: SARS-CoV-2 antibody responses do not predict clinical outcome in hospitalized patients with moderate-to-severe COVID-19 infection.

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Background: The direct effect of SARS-CoV-2 on the lungs results in increased hospitalization rates of patients with pneumonia. Severe COVID-19 patients often develop ARDS which is associated with poor prognosis. Assessing risk factors for COVID-19 severity is indispensable for implementing and evaluating therapeutic interventions. We investigated the temporal associations between the SARS-CoV-2 antigen (Ag), total Immunoglobulin (Ig) levels, and several laboratory parameters in hospitalized patients with varying degrees of COVID-19 severity. Methods: The SARS-CoV-2 nucleocapsid protein (NP) and total Ig Spike (S) protein-specific antibodies were determined for each patient with lateral flow assays through repeated sampling every two days. Hematological and biochemical parameters were evaluated at the same time points. Results: 40 Greek COVID-19 patients (31 males, 9 females) with a median age of 59.50 ± 16.21 years were enrolled in the study. The median time from symptom onset to hospitalization was 8.0 ± 4.19 days. A significant negative correlation was observed between the SARS-CoV-2 Ag and total Ig levels. The temporal correlation patterns of the SARS-CoV-2 NP Ag and anti-S total Ig levels with laboratory markers varied among patients with differing degrees of COVID-19 severity. Severe-critical cases had lower SARS-CoV-2 Ag and increased total Ig levels as compared to mild-moderate cases. Conclusions: Distinct temporal profiles of the SARS-CoV-2 NP Ag and anti-S total Ig levels may distinguish different groups of COVID-19 severity.

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BACKGROUND: We aimed to estimate the SARS-CoV-2 antigen and antibody seroprevalence in one of the worst-affected by the pandemic semi-closed communities in Greece, Deskati, and evaluate the sociodemographic and clinical correlations of functional antibody responses. METHODS: The Ag2019n-CoV V1310/V1330 Rapid Test (Prognosis Biotech, Greece) was used for antigen detection. The Rapid Test 2019-nCoV Total Ig, V1210/V1230 (Prognosis Biotech, Greece), and the SARS-CoV-2 IgG II Quant method (Architect, Abbott, Illinois, USA) were used for antibody testing. RESULTS: None of the participants had a positive antigen result. SARS-CoV-2 seropositivity ranged from 13% to 45% in the study population, depending on the method. One-third of the participants with known past infection had a positive antibody test result 77 ± 13 days after infection. Two-fifths of infections determined by serology were asymptomatic. The advancing age and hospitalization predicted seropositivity among patients with past infection. Half of the participants who tested positive for antibodies were not aware of past infection. CONCLUSIONS: High-burden contexts in Greece, such as Deskati, are not so far from herd immunity thresholds. We highlighted the value of low-cost serosurveys targeting both symptomatic and asymptomatic populations to evaluate the natural immune response to SARS-CoV-2 in nonvaccinated susceptibles and design evidence-based policies for lifting lockdowns.

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Extracellular vesicles (EVs) are cell-secreted, lipid membrane-enclosed nanoparticles without functional nucleus. EV is a general term that includes various subtypes of particles named microvesicles, microparticles, ectosomes or exosomes. EVs transfer RNA, DNA and protein cargo between proximal and distant cells and tissues, thus constituting an organism-wide signal transduction network. Pathological tissues secrete EVs that differ in their cargo composition compared to their healthy counterparts. The detection of biomarkers in EVs from biological fluids may aid the diagnosis of disease and/or monitor its progression in a minimally invasive manner. Among biological fluids, pleural effusions (PEs) are integrated to clinical practice, as they accompany a wide variety of lung disorders. Due to the proximity with the pleura and the lungs, PEs are expected to be especially enriched in EVs that originate from diseased tissues. However, PEs are among the least studied biofluids regarding EV-specialized isolation methods and related biomarkers. Herein, we describe a practical EV isolation method from PEs for the screening of EV RNA biomarkers in clinical routine. It is based on a Proteinase K treatment step to digest contaminants prior to standard polyethylene-glycol precipitation. The efficiency of the method was confirmed by transmission electron microscopy, nanoparticle tracking analysis and Western blot. The reliability and sensitivity of the method towards the detection of EV-enriched RNA biomarkers from multiple PEs was also demonstrated.

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