Time-Resolved Fluorescence Spectroscopy of Blood, Plasma and Albumin as a Potential Diagnostic Tool for Acute Inflammation in COVID-19 Pneumonia Patients.

Fluorescence lifetime measurements of blood or plasma offer valuable insights into the microenvironment and molecular interactions of fluorophores, particularly concerning albumin. Neutrophil- and hypoxia-induced oxidative stress in COVID-19 pneumonia patients leads to hyperinflammation, various oxidative modifications of blood proteins, and potential alterations in the fluorescence lifetime of tryptophan-containing proteins, especially albumin. The objective of this study was to investigate the efficacy of time-resolved fluorescence spectroscopy of blood and plasma as a prompt diagnostic tool for the early diagnosis and severity assessment of COVID-19-associated pneumonia. This study examined a cohort of sixty COVID-19 patients with respiratory symptoms. To investigate whether oxidative stress is the underlying cause of the change in fluorescence lifetime, human serum albumin was treated with chloramine T. The time-resolved spectrometer Life Spec II (Edinburgh Instruments Ltd., Livingston, UK), equipped with a sub-nanosecond pulsed 280 nm diode, was used to measure the fluorescence lifetime of blood and plasma. The findings revealed a significant reduction in the fluorescence lifetime of blood (diluted 200 times) and plasma (diluted 20 times) at 360 nm in COVID-19 pneumonia patients compared with their respective values recorded six months post-infection and those of healthy individuals. Significant negative correlations were observed between the mean fluorescence lifetime of blood and plasma at 360 nm and several severity biomarkers and advanced oxidation protein products, while a positive correlation was found with albumin and the albumin-globulin ratio. The time-resolved fluorescence spectroscopy method demonstrates the potential to be used as a preliminary screening technique for identifying patients who are at risk of developing severe complications. Furthermore, the small amount of blood required for the measurements has the potential to enable a rapid fingerstick blood test.

The Impact of COVID-19 on Cellular Factors Influencing Red Blood Cell Aggregation Examined in Dextran: Possible Causes and Consequences.

Several studies have indicated that COVID-19 can lead to alterations in blood rheology, including an increase in red blood cell aggregation. The precise mechanisms behind this phenomenon are not yet fully comprehended. The latest findings suggest that erythrocyte aggregation significantly influences microcirculation, causes the formation of blood clots in blood vessels, and even damages the endothelial glycocalyx, leading to endothelial dysfunction. The focus of this research lies in investigating the cellular factors influencing these changes in aggregation and discussing potential causes and implications in the context of COVID-19 pathophysiology. For this purpose, the aggregation of erythrocytes in a group of 52 patients with COVID-19 pneumonia was examined in a 70 kDa Dextran solution, which eliminates the influence of plasma factors. Using image analysis, the velocities and sizes of the formed aggregates were investigated, determining their porosity. This study showed that the process of erythrocyte aggregation in COVID-19 patients, independent of plasma factors, leads to the formation of more compact, denser, three-dimensional aggregates. These aggregates may be less likely to disperse under circulatory shear stress, increasing the risk of thrombotic events. This study also suggests that cellular aggregation factors can be responsible for the thrombotic disorders observed long after infection, even when plasma factors have normalized. The results and subsequent broad discussion presented in this study can contribute to a better understanding of the potential complications associated with increased erythrocyte aggregation.

Trends from the Last Decade with Nontuberculous Mycobacteria Lung Disease (NTM-LD): Clinicians' Perspectives in Regional Center of Pulmonology in Bydgoszcz, Poland.

BACKGROUND: Nontuberculous mycobacteria (NTM) are the cause of chronic lung disease called NTM lung disease (NTM-LD). There are about 180 known species of NTM. Nowadays the number of NTM-LD is increasing. OBJECTIVE: To evaluate the clinical significance of NTM isolated from specimens and assess the frequency and clinical relevance of isolation of NTM in the Regional Center of Pulmonology in Bydgoszcz, hospital of Northern Poland. DESIGN: Clinical, radiological, and microbiological data were collected from all patients from whom NTM was isolated between 2013 and 2022. Data were reviewed retrospectively. Diagnostic criteria for NTM-LD published by the American Thoracic Society (ATS) were used to determine clinical relevance. MATERIAL AND METHODS: The study comprised 81,985 clinical specimens submitted for mycobacterial culture in the Department of Microbiology at the Regional Center of Pulmonology in Bydgoszcz between 2013 and 2022. Clinical specimens were processed according to the standard procedure in mycobacteria laboratories in Poland. NTM strains were identified using analysis of mycolic acids by chromatography as well as GenoType NTM-DR, GenoType Mycobacterium AS, and GenoType Mycobacterium CM. RESULTS: There were 395 patients with NTM strains between 2013 and 2022. Out of them, 149 cases met the diagnostic criteria of NTM-LD and were classified as definite cases. M. kansasii (n = 77) was the most common species in the group (51.68%), followed by M. avium complex (n = 46). Patients with NTM-LD were 22-88 years old (median age was 60 years). There were 81 men and 68 women. The most common symptoms were cough, hemoptysis, and fever. Radiological X-ray images were dominated by infiltrative lesions in the upper and middle lobe of the right lung with cavities; the changes were in the upper lobe of the left lung and on both sides of the chest. They were smokers in 61%. The most common concomitant diseases were chronic obstructive pulmonary disease (COPD), diabetes mellitus, pulmonary carcinoma, and human immunodeficiency virus (HIV) infection, and other immunodeficiencies. The most common treatment was isoniazid, ethambutol, rifampicin, and ofloxacin for 18 months with a minimum of 12 months of culture negativity. CONCLUSIONS: NTM-LD infections are present with other pulmonary illnesses and extrapulmonary diseases and may be connected to primary immunologic deficiencies. These diseases concern patients of all ages and have various clinical manifestations. M. kansasii and MAC are the most prevalent NTM isolates among respiratory samples in Northern Poland. In addition, an increase in MAC and a decrease in M. kansasii both in cultivation and the cause of NTM-LD were reported.

PCSK9 Inhibition During the Inflammatory Stage of SARS-CoV-2 Infection.

BACKGROUND: The intensity of inflammation during COVID-19 is related to adverse outcomes. Proprotein convertase subtilisin/kexin type 9 (PCSK9) is involved in low-density lipoprotein receptor homeostasis, with potential influence on vascular inflammation and on COVID-19 inflammatory response. OBJECTIVES: The goal of this study was to investigate the impact of PCSK9 inhibition vs placebo on clinical and laboratory outcomes in patients with severe COVID-19. METHODS: In this double-blind, placebo-controlled, multicenter pilot trial, 60 patients hospitalized for severe COVID-19, with ground-glass opacity pneumonia and arterial partial oxygen pressure to fraction of inspired oxygen ratio ≤300 mm Hg, were randomized 1:1 to receive a single 140-mg subcutaneous injection of evolocumab or placebo. The primary endpoint was death or need for intubation at 30 days. The main secondary endpoint was change in circulating interleukin (IL)-6 at 7 and 30 days from baseline. RESULTS: Patients randomized to receive the PCSK9 inhibitor had lower rates of death or need for intubation within 30 days vs placebo (23.3% vs 53.3%, risk difference: -30%; 95% CI: -53.40% to -6.59%). Serum IL-6 across time was lower with the PCSK9 inhibitor than with placebo (30-day decline: -56% vs -21%). Patients with baseline IL-6 above the median had lower mortality with PCSK9 inhibition vs placebo (risk difference: -37.50%; 95% CI: -68.20% to -6.70%). CONCLUSIONS: PCSK9 inhibition compared with placebo reduced the primary endpoint of death or need for intubation and IL-6 levels in severe COVID-19. Patients with more intense inflammation at randomization had better survival with PCSK9 inhibition vs placebo, indicating that inflammatory intensity may drive therapeutic benefits. (Impact of PCSK9 Inhibition on Clinical Outcome in Patients During the Inflammatory Stage of the COVID-19 [IMPACT-SIRIO 5]; NCT04941105).

Study of Albumin Oxidation in COVID-19 Pneumonia Patients: Possible Mechanisms and Consequences.

Oxidative stress induced by neutrophils and hypoxia in COVID-19 pneumonia leads to albumin modification. This may result in elevated levels of advanced oxidation protein products (AOPPs) and advanced lipoxidation end-products (ALEs) that trigger oxidative bursts of neutrophils and thus participate in cytokine storms, accelerating endothelial lung cell injury, leading to respiratory distress. In this study, sixty-six hospitalized COVID-19 patients with respiratory symptoms were studied. AOPPs-HSA was produced in vitro by treating human serum albumin (HSA) with chloramine T. The interaction of malondialdehyde with HSA was studied using time-resolved fluorescence spectroscopy. The findings revealed a significantly elevated level of AOPPs in COVID-19 pneumonia patients on admission to the hospital and one week later as long as they were in the acute phase of infection when compared with values recorded for the same patients 6- and 12-months post-infection. Significant negative correlations of albumin and positive correlations of AOPPs with, e.g., procalcitonin, D-dimers, lactate dehydrogenase, aspartate transaminase, and radiological scores of computed tomography (HRCT), were observed. The AOPPs/albumin ratio was found to be strongly correlated with D-dimers. We suggest that oxidized albumin could be involved in COVID-19 pathophysiology. Some possible clinical consequences of the modification of albumin are also discussed.

The Mortality Risk and Pulmonary Fibrosis Investigated by Time-Resolved Fluorescence Spectroscopy from Plasma in COVID-19 Patients.

A method of rapidly pointing out the risk of developing persistent pulmonary fibrosis from a sample of blood is extraordinarily needed for diagnosis, prediction of death, and post-infection prognosis assessment. Collagen scar formation has been found to play an important role in the lung remodeling following SARS-CoV-2 infection. For this reason, the concentration of collagen degradation products in plasma may reflect the process of lung remodeling and determine the extent of fibrosis. According to our previously published results of an in vitro study, an increase in the concentration of type III collagen degradation products in plasma resulted in a decrease in the fluorescence lifetime of plasma at a wavelength of 450 nm. The aim of this study was to use time-resolved fluorescence spectroscopy to assess pulmonary fibrosis, and to find out if the lifetime of plasma fluorescence is shortened in patients with COVID-19. The presented study is thus far the only one to explore the fluorescence lifetime of plasma in patients with COVID-19 and pulmonary fibrosis. The time-resolved spectrometer Life Spec II with the sub-nanosecond pulsed 360 nm EPLED® diode was used in order to measure the fluorescence lifetime of plasma. The survival analysis showed that COVID-19 mortality was associated with a decreased mean fluorescence lifetime of plasma. The AUC of mean fluorescence lifetime in predicting death was 0.853 (95% CI 0.735−0.972, p < 0.001) with a cut-off value of 7 ns, and with 62% sensitivity and 100% specificity. We observed a significant decrease in the mean fluorescence lifetime in COVID-19 non-survivors (p < 0.001), in bacterial pneumonia patients without COVID-19 (p < 0.001), and in patients diagnosed with idiopathic pulmonary fibrosis (p < 0.001), relative to healthy subjects. Furthermore, these results suggest that the development of pulmonary fibrosis may be a real and serious problem in former COVID-19 patients in the future. A reduction in the mean fluorescence lifetime of plasma was observed in many patients 6 months after discharge. On the basis of these data, it can be concluded that a decrease in the mean fluorescence lifetime of plasma at 450 nm may be a risk factor for mortality, and probably also for pulmonary fibrosis in hospitalized COVID-19 patients.

Analysis of the level of selected parameters of inflammation, circulating immune complexes, and related indicators (neutrophil/lymphocyte, platelet/lymphocyte, CRP/CIC) in patients with obstructive diseases.

INTRODUCTION: Chronic obstructive pulmonary disease (COPD) is an inflammatory disorder of the airways. An important element of COPD assessment is the evaluation of immune mechanisms involved in non-specific and specific response to ongoing inflammation. AIM OF THE STUDY: To evaluate the level of selected inflammatory and immunological parameters in patients with COPD, including C-reactive protein (CRP) and circulating immune complexes (CIC), as well as CRP/CIC index. MATERIAL AND METHODS: The study group consisted of 49 patients with obstructive pulmonary diseases (COPD, asthma, and asthma-COPD overlap syndrome) hospitalised in the Department of Pulmonary Diseases, Kuyavian-Pomeranian Pulmonology Centre in Bydgoszcz. Patients with COPD were divided into two subgroups, taking into account the severity of the disease according to the Global Initiative for Chronic Obstructive Lung Disease (GOLD; stages B and D). The control group consisted of 30 healthy persons. Levels of CIC were determined by the method of Hasková, and the concentration of CRP in serum by the standard immunoturbidimetric method. RESULTS: The median values of examined parameters (neutrophils, lymphocytes, platelets, neutrophil/lymphocyte ratio - NLR, platelet/lymphocyte ratio - PLR, CRP, CIC, and CRP/CIC index) were significantly higher among patients with obstructive diseases than in the control group. A tendency towards higher lymphocyte count, CRP, and CRP/CIC index in COPD stage D, compared to stage B, was observed. CONCLUSIONS: Based on our results, we suggest that the role of non-specific inflammatory mechanisms may increase in more advanced COPD stages (D), compared to less advanced stages (B).

[Smoking habit among parents of kids with respiratory system allergies during 10 years observation].

Allergies of the respiratory system are very often at children and are a global problem and still increasing. Passive smoking may predispose to allergies. Assuming that anti-smoking education conducted among of the children's parents during each control visit to the Allergy Clinic affects the behavior of the parents, we decided to analyze its effectiveness. Materials and Methods: The study comprised parents of 946 children at the Allergy Clinic, who were diagnosed and treated in years 2005-2014. The anti-nicotine education was applied by whole period of observation during routine medical visits. The outcome of an anti-smoking education achieved nearly 70 % efficiency. Results: Anti-nicotine education of the children's parents diseased on chronic allergic diseases of respiratory system is very good restrictive agent their exposition on smoking the tobacco. Contemporaneously in eftective way influences on decisions of adults about cessation smoking and the healthy style ot life promotes.

Unemployment in TB patients - ten-year observation at regional center of pulmonology in Bydgoszcz, Poland.

BACKGROUND: Tuberculosis (TB) affects the poorest of the poor and is an example of a disease that can contribute to the "disease-poverty trap". The variable epidemiological situation is associated with social risk factors, such as unemployment, which may favor the occurrence of this disease. The aim of this study was to analyze unemployment as a factor that can influence the incidence and course of the disease. MATERIAL AND METHODS: We analyzed TB patients with confirmed status of employment or unemployment admitted to the Regional Center of Pulmonology in Bydgoszcz in during the years 2001 to 2010. Out of 1130 patients, 604 were unemployed and the other confirmed their employment. RESULTS: The unemployed patients were mostly single men over age 40, with a low level of education, and living in a city. We observed that the proportions of smokers and alcohol abusers were significantly higher among the unemployed patients. The advanced radiological lesions, smear-positive pulmonary TB, and extra-pulmonary sites were diagnosed significantly more often in this group. The rate of death in the course of hospitalization was significantly higher in the group of unemployed patients. CONCLUSIONS: Unemployment among TB patients is a serious problem. We found that more advanced radiological lesions were associated with more frequent treatment interruptions and a higher rate of death in the course of hospitalization. Increased efforts are needed to reduce and eliminate the problem of unemployment among patients with TB. This may, indirectly, contribute to a decrease in notifications of TB cases and improve treatment outcomes.

[10 years comparative clinico-epidemiological analysis of smoking and alcohol consumption in TB patients (Myc. Tuberculosis) and with mycobacteriosis (Myc. Kansas]. / 10 letnia porównawcza analiza kliniczno-epidemiologiczna palenia i spoycia alkoholu u chorych na grulice (Myc. Tuberculosis) i mykobakterioe (Myc. Kansasi).

INTRODUCTION: Smoking and alcohol consumption are a major public health problem. More and more are mentioned, also, these two drugs, tobacco and alcohol as risk factors for tuberculosis and mycobacteriosis. AIM OF STUDY: Comparative analysis of epidemiological and clinical patients with tuberculosis and mycobacteriosis M.kansasii smoking cigarettes and abuse alcohol. MATERIAL AND METHOD: The study included 2025 patients with tuberculosis and 140 patients with diagnosed lung mycobacteriosis hospitalized in Kuyavian-Pomeranian Center of Pulmonology in the years 2003-2013. Data were obtained from the central database of the hospital on admission to the hospital. RESULTS: There were 1403 smokers (69.3%) of tuberculosis patients and 79 (56.4%) with mycobacteriosis, and alcohol dependence were 534 (26.4%) and 16 (11.4%) respectively. Both of smokers and drinkers, men prevailed. Smokers who have developed tuberculosis were significantly younger than patients with mycobacteriosis, often touched their homelessness and unemployment, and often lived in rural areas. Conversely, smokers with mycobacteriosis are people often married, professionally active. In the group of abusers, patients with tuberculosis were younger, living in the country. side, often unemployed, homeless and single compared to patients with my. cobacteriosis. The clinical picture of patients with tuberculosis and mycobacteriosis did not differ significantly between the groups. CONCLUSIONS: A retrospective study of patients with tuberculosis and my. cobacteriosis showed significantly more use of tobacco and alcohol abuse than in the general Polish population. It should be noted that cigarette smoking and alcohol abuse are major risk and mycobacteriosis. Therefore, it is important to conduct anti-tobacco education and prevention of alcohol abuse.

Diagnostic efficacy of myeloperoxidase for the detection of acute coronary syndromes.

BACKGROUND: Early diagnosis of acute coronary syndrome (ACS) is frequently a challenging task, while immediate risk stratification remains crucial for the prompt implementation of appropriate therapy in this setting. Employing markers that increase rapidly after the symptom onset may enhance triage and therapeutic decision-making in patients suspected for ACS. Myeloperoxidase (MPO) exerting proinflammatory and pro-oxidative properties is suggested as a reliable early marker for ACS associated with unfavourable clinical outcome. We assessed the diagnostic efficacy of plasma MPO alone or in combination with cardiac troponin I (cTnI) for detecting ACS in patients presenting with chest pain initiating within 6h before the hospital admission. MATERIAL AND METHODS: A study group consisted of 253 patients diagnosed with ACS and 47 subjects having other heart disease or unspecified chest pain. Clinically healthy volunteers (n=124) served as controls. MPO concentration was measured in plasma (Abbott Diagnostics, USA), while serum was assayed for cTnI, creatine-kinase MB, lipids, glucose, creatinine, brain natriuretic peptide type B and C-reactive protein. RESULTS: Both MPO and cTnI values were significantly lower in non-ACS subjects than in patients with ACS. At 97·5th percentile as cut-off, the superiority of MPO over cTnI was observed in patients with unstable angina and non-ACS subjects. Considerably higher MPO concentrations were demonstrated in the troponin-negative ACS patients on admission who became troponin-positive after 6h. Combined evaluation of MPO and cTnI possessed remarkably higher sensitivity than assessment of cTnI alone in all patients with ACS. CONCLUSIONS: Myeloperoxidase substantially facilitates the early diagnosis of ACS.

Multi-Marker Approach with the Use of Biochip Cardiac Array Technology for Early Diagnosis in Patients with Acute Coronary Syndromes.

INTRODUCTION: Diagnosis of acute coronary syndrome (ACS) is frequently a challenging task while immediate risk stratification remains crucial for the prompt implementation of appropriate therapy in this setting. The prolonged release pattern of both CK-MB mass and cardiac troponins makes it difficult to identify the origin of recent chest pain, thus a combination of early and later biomarkers might further facilitate the differential diagnosis. The study was designed to evaluate the efficacy of multi-marker approach using biochip array technology in identifying ACS shortly after the symptom onset. MATERIAL AND METHODS: The study group consisted of 42 patients suspected for ACS. Subjects were diagnosed as presenting with unstable angina (UA), non-ST-elevation myocardial infarction (NSTEMI) or ST-elevation myocardial infarction (STEMI). Biomarkers in the serum were determined twice: on admission (≤6 hours from the chest pain onset) and after next 6 hours. Cardiac troponin I was measured by routine sensitive automated assay (STAT cTnI) while other 6 cardiac markers (heart-fatty acid binding protein - H-FABP, myoglobin, glycogen phosphorylase BB, cTn I, CK-MB mass and carbonic anhydrase III) were assessed using biochip array technology. RESULTS: STAT cTnI concentrations within 6 hours from the symptom onset were elevated over the 99(th) percentile for reference population in 83.3% of subjects but none reached the cut-off value for myocardial infarction. Instead, H-FABP demonstrated a very good efficacy in early detection of ACS (90.5%), better than myoglobin and CK-MB mass. Sensitivity of H-FABP calculated for NSTEMI/STEMI subjects reached 100%. The diagnostic efficacy of troponin, myoglobin and CK-MB mass assay markedly increased within 12 hours. It was only for the patients with UA that the cardiac panel was not efficient in the early stratification of risk. CONCLUSIONS: A multi-marker strategy with H-FABP and highly sensitive troponin included enhances the early diagnosis and decision making process in patients with ACS. A new biochip cardiac array technology may serve as a powerful tool for ACS detection in the clinical practice.