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Aim. To evaluate the changes of cell-free DNA (cfDNA) levels before and after percutaneous coronary intervention (PCI) in patients with ST-segment elevation acute coronary syndrome (STE-ACS). To identify associations of cfDNA concentration before and after PCI with complications and length of ulcerated plaque in patients with STE-ACS.Material and methods. This prospective single-center observational pilot study included 44 patients with STE-ACS admitted to the cardiac intensive care unit during the period of May-August 2023. In all patients, along with standard laboratory tests, cfDNA level was measured upon admission and 24 hours after PCI. Assessment of cfDNA associations before and after PCI was carried out in relation to following significant complications and conditions in STE-ACS patients: death, acute left ventricular failure (ALVF), acute heart failure (AHF), arrhythmia, number of stents implanted, number of segments with impaired local contractility, total stent length.Results. The mean age of the patients was 60,6±9,6 years, of which 74,6% were men. TIMI 0-1 flow was recorded in 93,2% of the subjects. The most common complications were cardiogenic shock (18,4%), arrhythmia (16,9%), AHF (13,6%), ALV (11,9%). Death was recorded in 8,5%. Implantation of 1 stent in PCI was performed in 75% of cases, while in the rest, 2 or more stents were implanted. The proportion of patients with impaired local contractility was 90%, the median stent length was 24,0 (20,0-50,0) mm. CfDNA level on admission did not differ from level after PCI 94,5 (78,3-155,5) ng/ml vs 115,0 (71,0-152,0), p=0,46. However, it signi­ficantly exceeded the cfDNA concentration from a group of healthy volunteers (78,0 (59,7-106,0), p=0,017). Characteristic curve showed significant relationships both for the concentration of cfDNA before (with implantation of 2 or more stents (AUC 0,71 with 95% confidence interval (CI) 0,56-0,86, p=0,039), stent length >24 mm (AUC 0,73 with 95% CI 0,58-0,89, p=0,009)) and after PCI (with the number of impaired local contractility segments (AUC 0,73 with 95% CI 0,57-0,89, p=0,014)). If the cfDNA level before PCI was >90 ng/ml, the risk of implantation of 2 or more stents per procedure increased by 5,4 times (odds ratio (OR) 5,4, 95% CI 1,11-28,93, p=0,044). The risk of a stent length >24 mm with pre-PCI cfDNA >107 ng/ml increased 9-fold (OR 9,0 with 95% CI 2,2-36,9, p=0,001), and the cfDNA level after PCI >105 ng/ml increased the risk of impaired local left ventricular (LV) contractility in 2 or more segments by 5 times (OR 5,0, 95% CI 1,23-20,3).Conclusion. In the studied group of patients with STE-ACS subject to intervention, the cfDNA concentration before PCI was associated with the implantation of ≥2 stents and the stent length (>24 mm). CfDNA level before PCI was associated with the number of segments of impaired local LV contractility (≥2).

Purpose: to summarize available data on the diagnostic value of various circulating biomarkers for the detection of glioblastoma recurrence.Material and Methods. A literature search was conducted using PubMED ExoCarta and SILVA databases.Results. Glioblastoma multiforme (GBM) is the most common glioma in adults with an unfavorable prognosis. Treatment of tumor recurrence can improve the survival of patients. Neuroimaging is the standard method of diagnosing brain tumor recurrence. However, a neuroimaging method to clearly distinguish between pseudo progression and tumor progression has not been found to date. Current molecular tumor profling relies heavily on tissue resection or biopsy. Tissue profling has several disadvantages in the central nervous system’s tumors, including the challenge associated with invasive biopsy, the heterogeneous nature of many malignancies where a small biopsy can under represent the mutational profle. Liquid biopsy is a promising method in diagnosing malignant tumors. Blood collection is a simple, minimally invasive procedure, but cerebrospinal fuid allows tumor markers to be detected more confdently. However, collection of cerebrospinal fuid is a complex and invasive procedure that can be accompanied by serious complications.Conclusion. Biological fuid markers such as circulating tumor cells, extracellular vesicles, cell-free DNA and cell-free RNA allow for the detection of GMB, determination of molecular genetic features of cancer during response to therapy, and early detection of GBM recurrence.

ABSTRACT. Defining molecules with high prognostic value for predicting the course and outcomes of life-threatening sepsis, severe injuries, vascular accidents remains an urgent problem in emergency medicine. One of the promising candidate biomarkers of emergency states and critical illness is the content of extracellular DNA (exDNA) in blood plasma. The purpose of this review is to identify the prospects for the introduction of cfDNA in clinical medicine and the severities arose along this way. The levels and altered dynamics of the concentration of circulating DNA fragments, including the organ-specific fraction of exDNA seem informative today for assessing the degree of damage to the organ of interest, the probability of a complicated course and the prognosis of outcomes of emergency/critical illness in Intensive Care Unit (ICU) patients. Sources of exDNA circulating in the bloodstream may include the nuclei of dying cells from organs and tissues, damaged mitochondria, the pool of which should be remodeled with mitophagy, as well as microorganisms. Similarly to pathogen-associated molecules (PAMP) represented by fragments of bacterial and viral DNA, native DNA molecules associated with damage (DAMP) bind to toll-like receptors (TLR9) and intracellular DNA sensors (cGAS-STING, NLRP3), initiating the inflammatory processes in tissues and hemostatic disorders. These processes represent natural adaptive responses protecting against microbes, as well as disadaptation responses potentiating cell damage in organs. The increasing expression of genes encoding proinflammatory signaling pathways associated with NF-kB transcription factor and interferon-regulating factors (IRF), in turn, contribute to production of cytokines and other factors enhancing the stress-responses that alter the functional activity of cells in various organs. The available literature data suggest that the quantitative determining plasma exDNA, which serves as PAMP and DAMP to significantly contribute to pathogenesis of emergency states and critical illness, might aid in predicting the outcome and justifying the in-time personalization of treatment of emergency and post-emergency patients.

Purpose: to determine the informative value of the prediction of mortality of patients with sepsis using a combination of candidate markers — the quantitative content of circulating cell-free DNA (cfDNA) in plasma and allelic variants of TLR9 gene encoding-cellular recognition receptor TLR9 for DNA fragments.Materials and methods. The study included patients from five ICU of 4 four hospitals (n=156). The patients were divided into 2 groups: (1) with sepsis (based on SEPSIS-3, 2016, criteria), (n=81) and with acute cerebrovascular event (n=75). CfDNA was isolated from patient’s blood plasma using organic solvents and its concentration was established by a spectrophotometer using fluorescent intercalating agent SYBR Green binding DNA with high affinity. Genotyping of rs352162 allele variants of TLR9 gene was carried out with the help of polymerase chain reaction using tetraprimers followed by separation of products via electrophoresis and their visualization. The cfDNA concentrations were compared in patients differ in TLR9 rs352162 genotypes and outcome was predicted in 30 days after hospitalization using the ROC analysis.Results. As regards the cfDNA content, there was no significant difference between patients with sepsis (subgroups with abdominal or non-abdominal sepsis) from two different hospitals, and cfDNA concentration in groups of patients was not associated with the source of primary infection, which allowed increasing group size by pooling up the data. Pooled data have shown that the sensitivity and specificity of the lethal outcome prediction based on the cfDNA content depends on the genotype: in homozygotic patients withg TLR9 rs352162 CC genotype, the AUC was significantly higher than in the alternative group of patients with TLR9 rs352162 CT and TT genotypes.Conclusion. The data we obtained suggest high informative value of establishing the genetic polymorphism of TLR9 as complimentary to determining the cfDNA concentration and may demonstrate potential usefulness of selecting patients according to their TLR9 genotype for investigations of the clinical effect of targeted drugs inhibiting interaction of cfDNA with receptor TLR9.

The aim of the study was to improve approaches to the use of Rh0(D) immune globulin in pregnant women with Rh-negative blood by introducing noninvasive method of determination of fetal rhesus blood factor by mother’s blood into curation program of pregnancy at risk of RBC sensitization. The accuracy of noninvasive method of rhesus genotyping in 204 pregnant women with Rh negative blood in 14-27 weeks of pregnancy was 99.0%. The determination of fetal rhesus allows personify implementation of immunological prophylaxis in Rh negative pregnant women.

The discovery of the enormous role methylated cytosine plays in regulating gene expression has led to the development of a variety of techniques for detecting cytosine modification. A majority of these techniques are geared towards analyzing genomic DNA, which is typically available in large quantities. The concentration of cell-free DNAs (cfDNA) extracted from biological fluids including plasma, saliva, tears, or urine is relatively low and their degree of the fragmentation is high. Moreover, for noninvasive diagnostics of cancer, methylation patterns must be studied in minor cancer-specific fractions of DNA molecules substantially diluted by excess unmethylated molecules. The above limitations complicate the application of traditional techniques for cfDNA methylation analysis. In this manuscript, we review the state-of-art analysis of cfDNA methylation, hydroxymethylation, and noncanonical methylation (outside of CpG islands). The review covers methodological approaches to studying individual CpGs and genomic loci, as well as techniques for the large-scale analysis of methylation.