Here we report a pilot-sized study to compare the taxonomic composition of sputum microbiome in 17 newly-diagnosed lung cancer (LC) patients and 17 controls. Another object was to compare the ...representation of individual bacterial genera and species in sputum with the frequency of chromosomal aberrations in the blood lymphocytes of LC patients and in controls. Both groups were male; average age 56.1 ± 11.5 in patients and 55.7 ± 4.1 in controls. Differences in the species composition of bacterial communities in LC patients and controls were significant (pseudo-F = 1.94; p = 0.005). Increased prevalence in LC patients was detected for the genera Haemophilus and Bergeyella; whereas a decrease was observed for the genera Atopobium, Stomatobaculum, Treponema and Porphyromonas. Donors with high frequencies of chromosomal aberrations had a significant reduction in the microbiome of representatives of the genus Atopobium in the microbiome and a simultaneous increase in representatives of the species Alloprevotella compared to donors with a low level of chromosomal aberrations in lymphocytes. Thus, a comparison of the bacterial composition in the sputum of donors with cytogenetic damages in theirs lymphocytes, warrants further investigations on the potential role of microorganisms in the process of mutagenesis in somatic cells of the host body.
Severe course of COVID-19 is largely determined by hyperactivation of the immune system, or cytokine storm, in which immune cells (lymphocytes, monocytes,
etc
.) play a major role. Using low-voltage ...scanning electron microscopy, we studied the morphology of lymphocytes and monocytes during cytokine storm. Monocytes and lymphocytes were isolated by fluorescence sorting from the blood of healthy volunteers (
n
=6) and patients with COVID-19 (
n
=5) during cytokine storm (IL-6>23 ng/ml, smear positive for SARS-CoV-2). For each patient, 11-32 individual cells were analyzed at magnification of 18-32,000 times. Measurements showed that monocyte size was increased during cytokine storm (
p
=0.0001).
COVID-19 biobank: features of the cytokine profile Sushentseva, N. N.; Popov, O. S.; Apalko, S. V. ...
Kardiovaskuli͡a︡rnai͡a︡ terapii͡a︡ i profilaktika,
12/2020, Letnik:
19, Številka:
6
Journal Article
Recenzirano
Odprti dostop
Aim
.
Using a collection of samples from the biobank ofCityHospital № 40 ofSt. Petersburg, to study the cytokine profile in patients with coronavirus disease 2019 (COVID-19) and sepsis, in comparison ...with patients with abdominal inflammation and septicemia.
Material
and methods.
The study included serum samples from 181 patients with sepsis and COVID-19 (127 patients with a diagnosis confirmed by polymerase chain reaction (PCR); 54 patients with a negative PCR test, but with a characteristic computed tomographic lung performance) and 47 patients with abdominal sepsis. The content of cytokines was determined using a multiplex immunofluorescence analysis based on the Luminex xMAP technology using the HCYTOMAG60K panel — a soluble CD40 ligand (sCD40L), interleukin-1α (IL-1α), interleukin-1β (IL-1β), interleukin 6 (IL-6), interleukin 8 (IL-8), monocyte chemoattractant protein 1 (MCP-1), tumor necrosis factor alpha (TNFα), vascular endothelial growth factor (VEGF). Other laboratory parameters (C-reactive protein (CRP), ferritin, procalcitonin) were taken from patient records. Normality of distribution was assessed by the Shapiro-Wilk test. To compare groups, the Mann-Whitney test for independent samples, Wilcoxon test for dependent samples, and the Kruskal-Wallis test with Bonferroni correction for multiple comparisons were used.
Results
.
In patients with sepsis and COVID-19 infection, no differences in the concentrations of cytokines, ferritin and CRP were found between the groups with detected and not detected virus by PCR test. Based on this, this group was considered homogeneous when studying the cytokine profile. It was shown that in patients with sepsis and COVID-19, significantly higher levels of sCD40L (p<0,0001) and VEGF (p=0,037) and relatively low levels of CRP (p<0,0001), IL-6 (p<0,0001), IL-8 (p<0,0001), TNFα (p<0,00058).
Conclusion
.
These results indicate that sepsis in patients with COVID-19 courses with less elevation in inflammatory cytokine than in abdominal sepsis. At the same time, a critically high level of sCD40L indicates the significant endothelial damage.
Alzheimer's disease (AD) is the most common cause of dementia in the elderly. The aim of the work was
Objective:
to determine the relationship between laboratory biomarkers in blood plasma and ...cerebrospinal fluid (CSF) in patients with AD and indicators of neuropsychological testing.
Patients and methods.
52 patients with AD were examined, in which the concentration of 90 potential biomarkers were measured in blood plasma and CSF. Neuropsychological assessment included the Mini-Mental State Exam (MMSE), Frontal Assessment Battery (FAB), Montreal Cognitive Assessment (MoCA), and etc.
Results and discussion.
Correlations of different strength between the values of biomarkers in blood plasma and CSF and the results of neuropsychological assessment were revealed. A correlation was found between the soluble cell adhesion molecule (sICAM-1) in blood plasma and the largest number of neuropsychological tests sensitive to dementia stages (MoCA, MMSE, FAB) in patients with AD at the dementia stage. A correlation was found between the concentration of growth/differentiation factor 15 and interferon γ in blood plasma and FAB scores in patients with AD. The levels of granulocyte colony-stimulating factor (G-CSF) in CSF were associated with the dementia stage in AD, and the interleukin-1 receptor antagonist (IL-1RA) levels, on the contrary, with stages preceding the development of dementia in AD.
Conclusion.
sICAM-1 level in blood plasma, which is a marker of endothelial dysfunction, may be an indicator of the severity of the vascular neurodegenerative process in AD at the dementia stage. G-CSF in the CSF is associated with the dementia stage in AD, and IL-1RA – with the pre-dementia stage of AD, which determines the prospect of their further study as diagnostic markers.
Aim.
To optimize the technique for the isolation and storage of ribonucleic acid (RNA) from whole blood and leukocyte fraction.
Materials and methods
. Comparison of isolation quality was carried out ...for RNA samples obtained from 228 leukocyte samples and 198 whole blood samples. Isolation was performed from fresh and frozen samples using ExtractRNA™ reagent and a MagNA Pure Compact automated system. Various methods of removing erythrocytes (centrifugation and treatment with hemolytic agents from two manufacturers) were tested, as well as freezing with and without preservatives for subsequent RNA isolation.
Results.
Twenty-one combinations of conditions were tested. The highest quality RNA was isolated by manual extraction using the ExtractRNA™ reagent from a fresh leukocyte fraction, purified by the Amplisens hemolytic agent (successful extraction — 94%, median RIN=8,4); frozen in IntactRNA™, purified by leukocyte fraction centrifugation (successful extraction — 100%, median RIN=8); frozen in ExtractRNA™, purified by leukocyte fraction centrifugation (successful extraction — 100%, median RIN=9,3).
Conclusion.
RNA can be isolated from frozen blood fractions, which is not inferior in quality to that isolated from fresh samples. Thus, it is not necessary to isolate RNA immediately after the receipt of biological material.
Secretory phospholipases A2 (sPLA2) represent a large superfamily of enzymes with a molecular weight of 14-19 kDa, including 15 groups and more than 30 isoforms belonging to four types: secretory ...(sPLA2), cytosolic (cPLA2), calcium-independent (iPLA2) and lipoprotein-associated phospholipase A2 (LP-PLA2, PAF-AH). Eleven species of secretory sPLA2s (IB, IIA, IIC, IID, IIE, IIF, III, V, X, XIIA, and XIIB) have been found in mammals, performing versatile functions and participating in the pathogenesis of a wide range of diseases. On the one hand, sPLA2 may promote elimination of damaged, apoptotic cells by hydrolyzing membrane phospholipids, and exerts a strong bactericidal and antiviral properties, including pronounced effects against antibiotic-resistant strains of microorganisms. In this regard, the use of sPLA2 may represent a new strategy for the treatment of bacterial and viral infections. Moreover, due to the action of sPLA2 on its substrates, a number of biologically active molecules (arachidonic, lysophosphatidic acids, lysophospholipids, fatty acids, prostaglandins, leukotrienes, thromboxanes) are formed, which provide strong inflammatory, detergent, coagulating effects and increase vascular permeability. This pro-inflammatory role of sPLA2 may explain its increase levels and activity in cardiovascular, respiratory, autoimmune, metabolic, oncological, bacterial and viral disorders. The review article presents a classification of sPLA2 isoforms, their substrates, regulatory factors, biological significance, and mechanisms of their strong bactericidal, virucidal, and pro-inflammatory activity in the heart and lung disorders, autoimmune, metabolic, bacterial, and viral diseases. In particular, the mechanisms of the selective action of sPLA2 against Gram-positive and Gram-negative microorganisms are discussed. We consider diagnostic and prognostic significance, correlations between elevated levels and activity of sPLA2 and distinct clinical symptoms, severity and outcome in the patients with coronary heart disease (CAD), acute myocardial infarction (AMI), atherosclerosis, acute inflammatory lung injury (ALI), respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD), rheumatoid arthritis, bronchial asthma, bacterial infections, septicemia and viral (COVID-19) infections. The opportunity of using sPLA2 as a biomarker of the severity and outcome of patients with chronic obstructive pulmonary disease, bacterial infections, sepsis and viral infections, including COVID-19, is also considered.
Aim
. To determine the reference ranges of concentrations of 38 cytokines, chemokines and growth factors, as well as to measure the content of these analytes in patients with neurodegenerative and ...cardiovascular diseases (CVDs) using biomaterial from the biobank repository.
Material and methods
. The study included 303 serum and plasma samples from 281 healthy donors, 242 samples from 224 patients with neurodegenerative diseases, and 164 samples from 152 patients with CVDs from the biobank of the of St. Petersburg City Hospital № 40. In all samples, the concentration of 38 cytokines, chemokines, and growth factors was determined by multiplex immunofluorescence assay.
Results
. Based on the measured concentrations in the group of healthy donors, non-parametric 95% reference ranges with 90% confidence intervals were calculated. For the majority of analytes, no sex and age differences were observed. In donors >65 years of age, the concentration of macrophage inflammatory protein-1-α was reduced and the levels of interleukin-8 and the chemokine interferon-inducible protein 10 were increased. Young donors (18-35 years) had lower levels of tumor necrosis factor-α. In groups with neurodegenerative and cardiovascular diseases, multiple deviations from the calculated reference values were found.
Conclusion
. Certain reference intervals are intended to evaluate the concentrations of cytokines, chemokines and growth factors determined in blood serum or plasma using Human Cytokine/Chemokine Magnetic Bead Panel 1 reagents (Merck, Millipore) on the MAGPIX system and do not imply a direct transfer to other analytical methods.
Aim.
To analyze the metabolomic profile of patients with coronavirus disease 2019 (COVID-19) using mass spectrometry methods and taking into account clinical and laboratory history. Studying the ...relationship between the severity of COVID-19 symptoms and the concentration of primary metabolites, primarily amino acids.
Material and methods
. The retrospective study included serum samples from a total of 935 people (445 men and 490 women) with COVID-19 from the collection of the biobank of the City Hospital № 40. A metabolomic profile was studied in all samples using chromatography-mass spectrometry. The R language was used for statistical data processing.
Results.
Based on the analysis, a difference in the metabolic profile of patients with COVID-19 was identified depending on the disease severity — for 52 of the 84 detected compounds, differences were observed with a significance of p<0,01. Significant differences in concentration were recorded for organic acids, amino acids and their derivatives.
Conclusion.
Using samples from the biobank collection, a metabolomic study of the biomaterial of patients hospitalized with a diagnosis of COVID-19 was carried out. According to the results obtained, kynurenine, phenylalanine and acetylcarnitine are associated with COVID-19 severity. Differences in the metabolomic profile were identified in patients with cytokine storm at the time of blood collection. Potential biomarkers for cytokine storm were identified.
The prevalence of metabolic, vascular and hepatic disorders in young and middle-aged men suffering from abdominal obesity was estimated. It was found that in men suffering from initial obesity, ...compared with patients with normal diet, a high frequency of arterial hypertension (67 and 37%, χ2=41,5; p
A critical role in coronavirus disease 2019 (COVID-19) pathogenesis is played by immune dysregulation that leads to a generalized uncontrolled multisystem inflammatory response, caused by ...overproduction of proinflammatory cytokines, known as "a cytokine storm" (CS), strongly associated with a severe course of disease. The aim of this study is to identify prognostic biomarkers for CS development in COVID-19 patients and integrate them into a prognostic score for CS-associated risk applicable to routine clinical practice.
The authors performed a review of 458 medical records from COVID-19 patients (241 men and 217 women aged 60.0 ± 10.0) who received treatment in the St. Petersburg State Budgetary Institution of Healthcare City Hospital 40 (City Hospital 40, St. Petersburg), from Apr. 18, 2020 to Nov. 21, 2020. The patients were split in two groups: one group included 100 patients with moderate disease symptoms; the other group included 358 patients with progressive moderately severe, severe, and extremely severe disease. The National Early Warning Score (NEWS) score was used alongside with clinical assessment, chest computed tomographic (CT) scans, electrocardiography (ECG), and lab tests, like ferritin, C-reactive protein (CRP), interleukin (IL)-6, lactate dehydrogenase (LDH), and D-dimer.
The basic risk factors for cytokine storms in COVID-19 patients are male gender, age over 40 years, positive test result for replicative severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA, absolute lymphocyte count, dynamics in the NEWS score, as well as LDH, D-dimer, ferritin, and IL-6 levels. These clinical and instrumental findings can be also used as laboratory biomarkers for diagnosis and dynamic monitoring of cytokine storms. The suggested prognostic scale (including the NEWS score dynamics; serum IL-6 greater than 23 pg/ml; serum CRP 50 mg/L or greater; absolute lymphocyte count less than 0.72 × 10
/L; positive test result for replicative coronavirus (SARS-CoV-2) RNA; age 40 years and over) is a useful tool to identify patients at a high risk for cytokine storm, requiring an early onset of anti-inflammatory therapy.
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