The main results of the Scientific and Technical Program of the Union State “DNA Identification” and implementation of reagent kits and technologies developed under the Program to establish the ethnogeographical and population origin of an unknown individual on the basis of his DNA are presented. The developed reagent kits are designed for DNA identification and are adapted to existing national reagents and their instrumentation used in Union State forensic laboratories.

Objective. To study the clinical and anamnestic features in patients with various forms of fetal stunting to determine the risk factors for the development of this pregnancy complication. Material and methods. The study included 329 patients, who, depending on pregnancy outcomes, were divided into 3 groups: The 1st group (main group) included 165 women whose pregnancy was complicated by insufficient fetal growth (IFG). The main group was divided into 2 subgroups: 1a — 72 patients, whose pregnancy was complicated by the development of a fetus of low gestational age (LGA), 1b — 93 patients, whose pregnancy was complicated by fetal growth retardation. The 2nd group (comparison) included 164 patients whose pregnancy ended with the birth of a live full-term baby without signs of insufficient growth. The somatic and obstetric anamnesis and the course of pregnancy were studied in all patients. Results. It was shown that patients younger than 20 years of age were more likely to occur in the SGA and FGR subgroups (odds ratio — OR 5.8; 95% confidence interval — CI 1.450—23.039; p=0.010 and OR 4.4; 95% CI 1.102—17.322; p=0.039, respectively). An analysis of the maternal history showed that preterm birth was more common in the SGA group than in the comparison group (OR 2.8; 95% CI 1.077—7.165; p=0.038). Insufficient fetal growth in the previous pregnancy was more common in women of the main group and increased the chance of this pregnancy complication in the present pregnancy: 4.3 times (OR 4.3; 95% CI 1.282—14.330; p=0.016) for FGR and 3.7 times (OR 3.7; 95% CI 0.994—13.306; p=0.072) for a fetus of low weight for this gestational age. Uterine scarring was more common in patients of the SGA group than in the comparison group (OR 2.4; 95% CI 1.143—4.867; p=0.029). Early toxicosis and gestational diabetes mellitus were more common in patients in the FGR group than in the comparison group (OR 3.1; 95% CI 1.622—6.011; p<0.001 and OR 2.1; 95% CI 1.181—3.829; p=0.014). Conclusion. The study identified new risk factors for breast undergrowth. It is necessary to take into account the patient’s age, the presence of a scar on the uterus, premature birth, insufficient fetal growth in a previous pregnancy, early toxicosis, and gestational diabetes mellitus in order to timely assess the risk of insufficient fetal growth.

Aim. To evaluate the role of biochemical and biophysical parameters in the combined first-trimester prenatal screening for the development of clinical forms of fetal growth insufficiency. Materials and methods . Group I (main) included 73 patients, whose pregnancies were complicated by the fetal growth insufficiency. The main group was divided into two subgroups: Ia with 30 patients whose pregnancies were complicated by fetal growth restriction (FGR) and Ib with 43 patients whose pregnancies were complicated by small for gestational age fetuses (SGA). Group II (control) included 118 patients whose pregnancies resulted in the birth of a live, full-term infant with normal height and weight. All patients underwent combined first-trimester prenatal screening with calculation of biochemical (pregnancy-associated plasma protein A (PAPP-A), free β-subunit of human chorionic gonadotropin (β-hCG) and biophysical (mean arterial pressure (MAP), uterine artery pulsatility index (PI) parameters, the values of which were subsequently analyzed. Results. The level of PAPP-A was statistically significantly lower in the FGR group (0.793 MoM) compared to the control group (1.048 MoM), p = 0.005. The level of PAPP-A in the blood below 0.793 MoM increases the risk of fetal growth restriction by 3.244 times (odds ratio (OR) = 3.244; 95% confidence interval (95% CI) 1.394–7.554, p = 0.005). An increase in the pulsation index was found in Doppler ultrasound of the uterine arteries in patients with FGR compared to the SGA group (OR = 2.254; 95% CI 0.990–5.129, p = 0.017). Statistically significant differences were not found in the studied parameters of the combined first-trimester prenatal screening in relation to the development of SGA. Conclusion. Differences in the biochemical and biophysical parameters of combined prenatal screening for the clinical forms of the fetal growth insufficiency were identified. Further research is needed to identify new prognostic markers of fetal growth insufficiency, which will help reduce perinatal losses. Additional research is required to expand the sample size of the Russian population to clarify the role of the prenatal screening components.

Fluorescent in situ hybridization (FISH) remains an indispensable tool for molecular diagnostics, which makes it possible to detect chromosomal abnormalities underlying many hereditary and oncological diseases with high accuracy. The advancement of medicine towards personalized approaches and the expansion of the spectrum of diagnosed pathologies require constant improvement of methods for synthesizing DNA probes. Despite existing limitations such as the cost and complexity of synthesis, the future of FISH diagnostics is linked to the development of highly specific, multiplex, and affordable probes that will enable the transition to complex genome and transcriptome analysis. The aim of this article was to reflect the evolution of probe production methods from classical to high-tech, including SABER-FISH, CRISPR/Cas9 (CASFISH), and smFISH technologies.

COVID-19 is a severe acute respiratory infection caused by the SARS-CoV-2 virus. Research in the field of host genetics contributes to the discovery of new genomic markers of coronavirus infection progression. In this article, a method for multiplex genotyping of polymorphic variants of genes associated with the severity of COVID-19 has been developed, based on multilocus PCR and MALDI-TOF mass spectrometry of DNA molecules. The frequencies of 45 single nucleotide polymorphisms of COVID-19 candidate genes in a population sample of Russians from Tomsk are characterized. The results are compared with data for populations from the 1000 Genomes Project.

Аневризма аорты (АА) и атеросклероз (АС) характеризуются неоднозначностью коморбидных отношений между собой. В настоящем обзоре рассматриваются молекулярные механизмы формирования данных патологий, обусловленные гетерогенностью, пластичностью, межклеточными взаимодействиями, эмбриональным происхождением и региональной специфичностью клеток артерий, выявляемыми с помощью подхода транскриптомики отдельных клеток у человека и на модельных животных. В результате подчеркивается важность взаимодействия генетических и средовых факторов, определяющих функциональное состояние сосудов и развитие патологии через динамическое изменение клеточного состава артерий в рамках онтогенетически регулируемого пространственно-временного континуума, что создает условия формирования коморбидности между заболеваниями. Понимание ключевых молекулярных механизмов, лежащих в основе коморбидности АА и АС, важно для разработки новых терапевтических стратегий данных патологических состояний.

Numerous histological studies have demonstrated that impaired placentation processes are involved in the key pathogenetic mechanisms of great obstetrical syndromes (GOSs). However, the molecular basis of this discovery is still unclear. Therefore, the objectives of this work were to characterize the molecular mechanisms and to search for new genetic markers of pregnancy complications via an integrative analysis of the data obtained by genome-wide expression profiling of placental tissue in preeclampsia, intrauterine growth restriction (IUGR), premature labor (PL), and physiological pregnancy (PP). Oxidative stress, ferroptosis, and disordered intercellular interactions in placenta were assumed to be common pathogenetic mechanisms of GOSs. A total of 64 genes were found to be significantly dysregulated in at least two pregnancy complications. Maternal endothelial cells and syncytiotrophoblast cells were the most significant cell populations enriched in these genes. A computational analysis and the topology of the protein–protein interaction network identified SOD1, ACTG1, TXNRD1, TKT, GCLM, GOT1, ACO1, and UBB as hub genes. A set of key regulators that trigger the reaction cascades involving the differentially expressed genes was found to include MAPK3, MID1, LCMT1, DUSP10, TOPS, SOX10, EGFR, TFAP2A, GLIS1, NR2F1, NR2F2, PAX5, HSF1, and BCL6. The genes were overrepresented in the MAP kinase and interferon-γ response signaling pathways. The above genes and their products were assumed to provide the most promising biomarkers for developing new approaches to risk factor assessment and targeted therapy in GOSs. Further studies should be aimed at clarifying their functional and diagnostic significance in pregnancy complications.

Objective. To evaluate the clinical features and the level of the inflammatory markers CCL5, slCAM-1, sVCAM-1, NCAM, PAI-1, and MPO in the groups of patients with Parkinson's disease (PD) at various stages according to Hoehn and Yahr. Material and methods. The study included 533 patients with PD. All patients underwent a clinical neurological examination to determine the stage of PD, the severity of motor disorders according to the MDS-UPDRS scale (Unified Parkinson's Disease Rating Scale of the Movement Disorder Society), and testing using validated questionnaires: Montreal Cognitive Assessment, Hospital Anxiety and Depression Rating Scale, Beck Depression Inventory-II, Fatigue Severity Scale, Scale for assessing autonomic disorders in PD patients. Behavioral disorders were evaluated using QUIP-RS. 144 PD patients had their serum concentration of several inflammatory markers measured (CCL5, slCAM-1, sVCAM-1, NCAM, PAI-1, and MPO) on the MAGPIX multiplex analyzer (Luminex, USA) using xMAP Technology. Genotyping of polymorphic variants of CCL5 (rs2107538) and PAI-1 (rs2227631) genes was performed using real-time PCR.

Results. The serum levels of slCAM-1, sVCAM-1, CCL5, and NCAM varied in PD patients depending on the Hoehn and Yahr stage and disease duration. Correlations of serum marker levels were found both among themselves and with motor and non-motor disorders, which indicate a systemic inflammatory profile when increased peripheral production of CCL5, slCAM-1, sVCAM-1, NCAM, PAI-1, and MPO may play a role in the neurodegenerative process.

Conclusion. The serum level of inflammatory markers, such as CCL5, slCAM-1, sVCAM-1, NCAM, PAI-1, and MPO, in PD patients varies depending on the stage of the progressive neurodegenerative process, indicating the importance of systemic inflammation during PD.

DYNC1H1 encodes the heavy chain of dynein 1, a protein that plays a critical role in intracellular transport and is also involved in neurogenesis, bipolar spindle apparatus formation, and interaction with certain regulatory proteins. Many variants of the gene are described in various neuromuscular, psychoneurological, congenital abnormalities, and malignancies. In this clinical case, the correlation of clinical manifestations with molecular genetic changes of the DYNC1H1 gene was evaluated. A variant was identified and presented de novo in the linker domain of the protein in a patient with abnormalities of brain development (pachygyria of the temporal and frontal lobes, polymicrogyria of the occipital lobes, cerebellar agenesis), polydactylitis, mental development disorder, and involvement of the neuromuscular system, as well as congenital cataracts. In this case, a new feature is described - polydactyly - not previously described in the variant c.4868G>A (p.Arg1623Gln), which expands the range of clinical manifestations and can contribute to the understanding of the mechanisms of phenotypic heterogeneity, as well as the development of optimal diagnostic algorithms.

Precocious puberty (PP, E30.1, Е22.8, Е30.9 according to ICD 10, MIM 176400, 615346) in children is a disorder in which secondary sexual characteristics appear earlier than the age norm. The timing of puberty is regulated by a complex interaction of genetic and epigenetic factors, as well as environmental and nutritional factors. This study aimed to search for pathogenic, likely pathogenic variants or variants of uncertain significance (VUS) in the KISS1, GPR54, DLK1, and MKRN3 genes in patients with the clinical picture of PP and normal karyotype by massive parallel sequencing. All identified genetic variants were confirmed by Sanger sequencing. The pathogenicity of identified genetic variants and the functional significance of the protein synthesized by them were analyzed according to recommendations for interpretation of NGS analysis results using online algorithms for pathogenicity prediction (Variant Effect Predictor, Franklin, Varsome, and PolyPhen2). Clinically significant genetic variants were detected in the heterozygous state in the KISS1R, DLK1, and MKRN3 genes in 5 of 52 probands (9.6 %) with PP, including 3 of 33 (9.1 %) in the group with central PP and 2 of 19 (10.5 %) in the group with gonadotropin-independent PP. Two children with gonadotropin-independent PP had VUS in the KISS1R gene (c.191T>C, p.Ile64Thr and c.233A>G, p.Asn78Ser), one of which was inherited from the father and the second, from the mother. The remaining patients with central PP had likely pathogenic genetic variants: DLK1:c.373delC(p.Gln125fs) de novo and DLK1:c.480delT(p.Gly161Alafs*49) of paternal origin. The third proband had a VUS variant in the MKRN3 gene (c.1487A>G, p.His496Arg), inherited from the father. All identified genetic variants were described for the first time in PP. Thus, in the present study, genetic variants in the KISS1R, DLK1, and MKRN3 genes in girls with PP were characterized

Цель. Оценить взаимосвязь дыхательной активности митохондрий лейкоцитов периферической крови с полиморфизмом митохондриальной ДНК (мтДНК) у пациентов с ишемической болезнью сердца (ИБС) в зависимости от наличия риска развития внезапной сердечной смерти (ВСС). Материалы и методы. Были сформированы две группы пациентов: основная группа – пациенты с ИБС и высоким риском ВСС (n = 107), группа сравнения – пациенты со стабильным течением ИБС без риска ВСС (n = 50). Пациентам определяли гаплогруппу, носительство полиморфизмов A2706G, G3010A и G9055A мтДНК методами высокопроизводительного секвенирования. Оценивали дыхательную активность изолированных митохондрий из лейкоцитов периферической крови амперометрическим методом при использовании NAD- и FAD-зависимых субстратов окисления. Результаты. В обеих исследованных группах гаплогруппы H, U, J являлись превалирующими (74,5 и 92,5% для основной группы и группы сравнения соответственно). В основной группе минорных гаплогрупп было больше, чем в группе сравнения. Частоты встречаемости полиморфизмов A2706G, G3010A, G9055A не имели значимых межгрупповых различий. В основной группе носительство замены A2706G ассоциируется со снижением коэффициента дыхательного контроля (ДК) при FAD-зависимом дыхании (р = 0,05), а в группе сравнения – со снижением скорости потребления кислорода (СПК) в метаболическом состоянии V4 при NAD- и FAD-зависимом типах дыхания (p = 0,002 и р = 0,008 соответственно) без изменения ДК. Носительство замены G9055A в основной группе ассоциировано со снижением СПК в метаболическом состоянии V3 (p = 0,037) при FAD-зависимом дыхании. Для полиморфизма G3010A мтДНК не выявлено связи с респираторной активностью митохондрий в исследованных группах. Заключение. У пациентов с ИБС вне зависимости от риска развития ВСС частоты гаплогрупп H, U, J и полиморфизмов A2706G, G3010A, G9055A мтДНК не имеют значимых различий. У пациентов высокого риска ВСС носительство полиморфизма A2706G связано с падением ДК при FAD-зависимом дыхании, а полиморфизма G9055A – со снижением СПК в V3 при FAD-зависимом дыхании.

Обоснование. Задержка роста плода остаётся одной из значимых патологий беременности, ассоциированной с высокой перинатальной и постнатальной заболеваемостью. Несмотря на прогресс в изучении молекулярных механизмов развития патологии, роль альтернативного сплайсинга ключевых плацентарных генов, в частности FLT1, кодирующего рецептор фактора роста эндотелия сосудов-1 (VEGFR-1), изучена недостаточно. Это ограничивает понимание связи между ангиогенезом и нарушениями развития плаценты при задержке роста плода. Цель исследования. Оценить роль альтернативного сплайсинга гена FLT1, экспрессируемого в децидуальных клетках плаценты, в молекулярных механизмах задержки роста плода. Методы. В исследование включены биоптаты материнской части плаценты пациенток с физиологической беременностью (n = 8) и задержкой роста плода (n = 13). Проведено секвенирование РНК на платформе Illumina NextSeq 2000. Анализ альтернативного сплайсинга выполнен с использованием пакета MAJIQ. Результаты. Впервые проведён анализ альтернативного сплайсинга гена FLT1 в децидуальных клетках плаценты при задержке роста плода. Общими для физиологической беременности и задержки роста плода являются четыре события альтернативного сплайсинга, включающие пропуск экзона, удержание интрона и два события, образующих комплекс. Кроме того, только при задержке роста плода выявлены одно комплексное событие и три события удержания интрона, отсутствующие при физиологической беременности. Суммарно, эти изменения отражают активацию механизмов, приводящих к формированию укороченных изоформ рецептора VEGFR-1, которые действуют как антиангиогенные «ловушки» и впоследствии приводят к снижению маточно-плацентарного кровотока и развитию задержки роста плода. Заключение. Альтернативный сплайсинг гена FLT1 играет важную роль в патогенезе задержки роста плода. Избыточное удержание интронов и пропуск экзонов ведут к повышенной экспрессии укороченных антиангиогенных белков, нарушая баланс ангиогенеза и способствуя плацентарной дисфункции.