Background. Chronic endometritis (CE) is a factor of endometrial receptivity impairment and one of the causes of recurrent implantation failure when using assisted reproductive technologies. Persistence of genital mycoplasmas and uterine cavity dysbiosis maintain chronic inflammation, which substantiates the need for antimicrobial therapy. Aim. To assess josamycin efficacy in patients with CE and recurrent implantation failure. Materials and methods. A total of 106 patients diagnosed with infertility and recurrent implantation failure were included in the study. A total of 33 patients with CE confirmed by immunohistochemistry and genital mycoplasmas in the endometrium and/or vagina only were selected for the study based on the examination results. Vaginal and endometrial microbiota was assessed by real-time polymerase chain reaction (Femoflor 16), and local inflammation was assessed based on cytokine expression (ImmunoQuantex) before and 1 month after josamycin therapy (500 mg 3 times a day, 10 days). Results. Eradication of genital mycoplasmas was achieved in 100% of cases. The decreased frequency of detecting facultative and obligate anaerobes in the uterine cavity was reported. A significant decrease in expression of interleukin-1β (p=0.0027), interleukin-18 (p=0.011), TLR4 (p=0.0489), along with the increase in the GATA3 transcription factor levels (p<0.001), was reported after treatment. The integrated assessment of the test cytokine transcript levels showed the decrease in the mean inflammation index from 86.2±18.7 to 45.2±0.6% (t=3.942, p=0.003) after the josamycin therapy course. Conclusions. The use of josamycin ensures complete eradication of genital mycoplasmas (Ureaplasma spp., Mycoplasma hominis, Mycoplasma genitalium), it is associated with the decrease in the prevalence of opportunistic microflora persistence in the uterine cavity, as well as with the decreased local inflammation severity. This makes it possible to regard it as a promising component of comprehensive CE treatment in patients with recurrent implantation failure.

BACKGROUND: Identifying the causes of spermatogenesis disorders has both medical and social importance. AIM: This study aimed to determine the relationship between testicular microcirculation parameters assessed by 3D ultrasound using VOCAL (Virtual Organ Computer-aided Analysis) software and ejaculate parameters in men from infertile couples. METHODS: This prospective, cross-sectional pilot study involved 24 men undergoing couple infertility examination. All men underwent semen analysis, testing for antisperm antibodies in the ejaculate, assessment of the proportion of spermatozoa with fragmented DNA using the TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labeling) assay, and scrotal ultrasound with 3D VOCAL visualization. The following parameters were assessed: testicular volume, vascularization index, flow index, vascularization-flow index, and mean signal intensity. RESULTS: The vascularization index and vascularization-flow index correlated significantly with the proportion of progressively motile spermatozoa (r = 0.42, p = 0.04; r = 0.43, p = 0.03, respectively) and significantly negatively with the proportion of immotile spermatozoa (r = −0.43, p = 0.04, r = −0.46, p = 0.02, respectively). CONCLUSION: This study is the first to use 3D angiography with VOCAL software to assess testicular microcirculation in men from infertile couples. This pilot study demonstrated the relationship between testicular blood supply and spermatogenesis parameters.

Background: Vaginal microbiota is a factor that determines a woman’s health. Infectious complications of pregnant women, women in labor and newborns are often associated with a significant change in its composition. Analysis of the species diversity of vaginal microbiota during pregnancy and the postpartum period primarily contributes to the study of physiological processes and the concept of a “healthy” vaginal environment during these significant periods of a woman’s life. Aim: The aim of this study was to evaluate the dynamic change in the species composition of vaginal microorganisms before delivery and at different times of the postpartum period in women with natural labor. Materials and methods: This study involved 24 pregnant women who delivered vaginally. Each patient was examined at three time points: 37–40 weeks of pregnancy (visit 1), 4–5 days of the postpartum period (visit 2), and 6–8 weeks after delivery (visit 3). The species composition of the vaginal microbiota was studied using a comprehensive test based on real-time quantitative polymerase chain reaction. Results: We found a decrease in the total bacterial mass in the vaginal biotope of women in labor after 4–5 days of the postpartum period and 6–8 weeks after delivery in comparison with the examination before delivery (p 0.0005). Similar changes were noted in the composition of the lactobacillary microbiota: the concentration of lactobacilli decreased in the postpartum period in comparison with that before delivery (p 0.05). Compared to the examination before delivery, 6–8 weeks after delivery in women, we observed a decrease in the frequency of Lactobacillus crispatus dominance (p 0.05), while the frequency of Lactobacillus iners dominance in the postpartum period increased (p 0.05). Among the representatives of the opportunistic vaginal microflora, the most frequently prevalent species were Gardnerella vaginalis, Prevotella bivia, Porphyromonas spp., Fannyhessea vaginae, Staphylococcus spp., Streptococcus spp., and Ureaplasma spp. In all women with severe vaginal microbiota disruption before delivery, an ascending bacterial infection was established according to the histological examination of the placenta after delivery (p 0.05). Conclusions: In most cases, the observed changes in the qualitative and quantitative composition of the vaginal microbiota in pregnant women and women in labor are physiological. At the same time, a consistent study of the vaginal microbiota during pregnancy and at different times of the postpartum period will allow for identifying possible risk factors for the development of infectious diseases in the mother and newborn and expand the possibilities for timely diagnosis and treatment of the identified disorders.

BACKGROUND: Fatty acids are important components of the oocyte microenvironment, exhibiting both pro-inflammatory and lipotoxic effects as well as anti-inflammatory effects depending on the presence and quantity of unsaturated bonds. Changes in the lipid profile of follicular fluid in obese patients may be a mechanism that leads to a decrease in oocyte competence and pregnancy rate in assisted reproductive technology programs. However, the available literature data is fragmentary and contradictory, which dictates the need for further research. AIM: The aim of this study was to investigate the lipid profile of follicular fluid in patients undergoing assisted reproductive technology programs with ovarian stimulation depending on the body mass index using MALDI (matrix-assisted laser desorption/ionization) mass spectrometry. METHODS: This study involved patients undergoing infertility treatment using assisted reproductive technology in a short protocol using gonadotropin-releasing hormone antagonists. The levels of essential saturated and unsaturated fatty acids were analyzed in the follicular fluid of the first aspirated follicle using MALDI mass spectrometry. RESULTS: Two study groups were formed out of 138 patients: patients with normal body mass index (n = 38) and patients with overweight and obesity (n = 76). The latter were characterized by higher levels of myristic and stearic acids and lower levels of oleic acid in the follicular fluid. The level of myristic acid in the follicular fluid positively correlated with the body mass index, while the level of oleic acid negatively correlated with this parameter. CONCLUSION: The data obtained indicate that overweight and obesity are associated with increased levels of saturated myristic and stearic acids in the follicular fluid, which negatively impact folliculogenesis, as well as a decrease in the level of monounsaturated oleic acid in the follicular fluid, which is able to compensate for the lipotoxic effects of saturated fatty acids.

BACKGROUND: Prevention of birth defects in children is a pressing issue. The total Down syndrome screening detection rate depends on the exact adherence to the prenatal examination algorithm and the average age of pregnant women. The introduction of new technologies increases the efficiency of chromosomal abnormality detection during total screening. AIM: The aim of this study was to assess the effect of organizational demographics on the frequency of births with chromosomal diseases, primarily Down syndrome, and to characterize additional factors to be taken into account when implementing a set of measures to prevent and avoid the birth of children with severe, uncorrectable and socially significant diseases. METHODS: This study included the results of a survey of 2,083 women in case of pre- and postnatal diagnosis of chromosome 21 trisomy in St. Petersburg, Russia from 2013 to 2023. Statistical data processing was carried out using standard software. RESULTS: We showed an increase in the average age of mothers and a decrease in the number of newborns in St. Petersburg over the specified period. The data on an increase in the efficiency of the prenatal diagnostics service were obtained, including using non-invasive prenatal testing for the group with an intermediate (from 1/101 to 1/1000) risk after combined screening. We also observed an increase in the timing of invasive diagnosis for prenatal karyotyping and the constant presence of a group of pregnant women who were not registered through city institutions and did not participate in preventive measures. CONCLUSION: From 2013 to 2023, the average incidence of Down syndrome (diagnosed pre- and postnatally) increased from 1/412 to 1/258 newborns. In general, the effectiveness of prenatal diagnosis increased to 94%; however, it is necessary to strive to transfer invasive diagnosis to earlier pregnancy. The average incidence of newborns with Down syndrome remains stable (1/1231).

In the present study, we aimed to investigate intratumoral karyotype diversity as well as the estrogen/progesterone effect on the cytogenetic profile of uterine leiomyomas (ULs). A total of 15 UL samples obtained from 15 patients were cultured in the media supplemented with estrogen and/or progesterone and without adding hormones. Conventional cytogenetic analysis of culture samples revealed clonal chromosomal abnormalities in 11 out of 15 ULs. Cytogenetic findings were presented by simple and complex chromosomal rearrangements (64% and 36% of cases, respectively) verified through FISH and aCGH. In most ULs with complex chromosomal rearrangements, the breakpoints did not feature clusterization on a single chromosome but were evenly distributed across rearranged chromosomes. The number of breakpoints showed a strong positive correlation with the number of rearranged chromosomes. Moreover, both abovementioned parameters were in a linear dependency from the number of karyotypically different clones per UL. This suggests that complex chromosomal rearrangements in ULs predominantly originate through sequential events rather than one hit. The results of UL cytogenetic analysis depended on the presence of estrogen and/or progesterone in the culture medium. The greatest variety of cytogenetically different cell clones was detected in the samples cultured without hormone supplementation. Their counterparts cultured with progesterone supplementation showed a sharp decrease in clone number, whereas such a decrease induced by estrogen or estrogen-progesterone supplementation was insignificant. These findings suggest that estrogen-progesterone balance is crucial for forming a UL cytogenetic profile, which, in turn, may underlie the unique response of the every karyotypically abnormal UL to medications.

Background: The role of the microbial factor in the genesis of early miscarriages (up to 10 weeks of pregnancy) has not been established. However, disruption of the vaginal microbiota may be associated with the occurrence of late miscarriage. Aim: The aim of this study was to evaluate the frequency of early and late miscarriages and to determine the prognostic significance of the microbial factor in early pregnancy registration. Materials and methods: This study involved 113 pregnant women registered before 10 weeks of gestation, who were divided into two study groups. Group I consisted of 22 women whose pregnancy ended in miscarriage before 22 weeks (subgroup IA of 19 patients with early miscarriage and subgroup IB of three patients with late miscarriage). Group II comprised 91 women whose pregnancy ended in term delivery. Vaginal microflora was investigated using microscopic and molecular biological methods. Results: The frequency of miscarriages among the examined women was 20.4%. The proportion of early miscarriages was 86.4% and late miscarriages 13.6%. In Group I, the number of births in the anamnesis was higher than in Group II. The average rate of births per woman was 2.0 in Group I and 0.9 in Group II (p = 0.037); the number of previous pregnancy loss per woman was 1.2 in Group I and 0.5 in Group II (p = 0.019). Lactobacilli were detected in the vaginal biotope of all pregnant women. Lactobacillus iners and Lactobacillus crispatus were the most frequently occurring species. In subgroup IA and Group II, opportunistic pathogenic microorganisms were detected with equal frequency at first pregnancy examination and did not alter the existing vaginal microbiocenosis. In subgroup IB, Gardnerella vaginalis, Atopobium vaginae, Enterobacteriaceae, Staphylococcus spp., Streptococcus spp., Mycoplasma hominis or Ureaplasma spp. were detected simultaneously in early pregnancy, with Lactobacillus cristatus predominating. Conclusions: The proportion of early miscarriages was six times higher than that of late miscarriages. We have not found an association between the quantitative and qualitative compositions of vaginal microorganisms in the first trimester of pregnancy and early miscarriages. The microbial factor was established to be important in late miscarriages. Further studies are needed to assess the prognosis of late miscarriages at the time of pregnancy registration.

BACKGROUND: Growth disturbances are common in pregnancies complicated by pregestational diabetes mellitus. Identifying the relationship between the structural and functional characteristics of the placenta and abnormal fetal growth is important for understanding its formation and the possibility of its prediction. AIM: The aim of this study was to conduct a comparative analysis of the postnatal morphological and prenatal ultrasound features of the structure of placentas in cases of fetal growth disorders in pregnant women with pregestational diabetes mellitus. METHODS: In this retrospective single-center cohort study, we analyzed the results of the morphological studies of 1200 placentas, including those with fetal growth disorders in pregnant women with pregestational diabetes mellitus. Ultrasound prenatal fetometry, placentometry and Doppler measurements were used in this study. The studied placentas were weighed, with their size and cotyledonous structure assessed. Placental histopathological parameters were diagnosed using standardized criteria. Statistical analysis was carried out using SPSS Statistics version 23.0. RESULTS: The comparison groups included patients with pregestational diabetes mellitus types 1 and 2 with the absence (n = 394) or the presence of various fetal growth disturbances such as fetal growth restriction (n = 109), small (n = 118) and large (n = 352) for gestational age fetuses. The control group (n = 157) consisted of pregnant women with normal fetal growth rates and normal carbohydrate metabolism. In the placentas from pregnant women with pregestational diabetes mellitus, regardless of the presence of fetal growth disorders, we identified a number of distinctive features compared to patients in the control group. These were abnormal size, inconsistency of the structure of the placenta and the gestational age with a predominance of dissociated villous maturation, the presence of circulatory disorders of varying degrees, the presence of inflammatory changes in the placenta, deposition of calcium salts, and the development of chronic placental insufficiency and placental infarction. Moreover, in pregestational diabetes mellitus, the placentas of fetuses with macrosomic and normal growth often demonstrated similar features of the morphological structure. In intrauterine growth restriction, signs of pathological immaturity, premature and abnormal maturation of the villi prevailed in the structure of the placenta, with sclerosis of the villous stroma, placental infarctions, and circulatory disorders being more common. We demonstrated an association of grade II and III hemodynamic disorders with the features of maturation and structure of the villi and the presence of placental insufficiency. Critical blood flow disorders in the umbilical artery were associated with severe circulatory disorders in the placentas. CONCLUSION: In analyzing the morphofunctional and ultrasound characteristics of the placenta in cases of fetal growth disorders in pregestational diabetes mellitus, we found changes associated with both impaired carbohydrate metabolism and the influence of concomitant conditions. Some features of placental morphology in pregestational diabetes mellitus appeared to be morphofunctional adaptations. A relationship was found between fetoplacental hemodynamics Doppler disturbances and the histological structure of the placentas.

Background/Objectives: This study reports on findings from the first preconception screening performed in Russia and provides a comprehensive discussion of the significant results and challenges faced during the implementation of the project. Methods: Using a targeted sequencing panel of 33 genes (associated with 29 autosomal recessive and 4 X-linked diseases), we analyzed 165 couples considering pregnancy. The screening design also included analysis of the frequent pathogenic variants in the SMN1, DMD, CFTR, and CYP21A2 genes that may not be detected through the next-generation sequencing approach. The sequential screening protocol, wherein the female partner was tested first, was used. Results: The results revealed that 35.8% of women (n = 59) were carriers of at least one pathogenic or likely pathogenic (P/LP) variant, with 7.9% of women (n = 13) carrying variants in two or more genes. Notably, the analysis identified 5 deletions of exon 7 in the SMN1 gene, 1 deletion of the CYP21A2 gene, and 1 large duplication in the DMD gene in female participants. The most frequently identified pathogenic variants occurred in the CYP21A2, GJB2, SERPINA1, and ATP7B genes. The screening identified six couples (3.6% of the cohort) at high risk of having a child with an autosomal recessive or X-linked genetic disorder. Conclusions: This pilot study confirms the high clinical utility of the gene panel, effectively evaluating reproductive risk in couples without a known family history of monogenic diseases. The findings indicate that the observed frequencies of identified gene variants differ from those theoretically expected, with a notable percentage of identified couples being at relatively high risk. Furthermore, these results highlight the indispensable role of comprehensive genetic counseling both before and after testing to ensure an appropriate preconception testing algorithm and informed reproductive decision-making.

Background: Chronic hyperglycemia and abnormal glucose variability are established risk factors for perinatal complications. Abnormal fetal growth and diabetic fetopathy can be associated not only with carbohydrate metabolism disorders, but also with impaired production of cytokines, chemokines, and growth factors that regulate intercellular interactions. Aim: The aim of this study was to evaluate cytokine and growth factor levels in the blood serum of pregnant women with type 1 diabetes mellitus and in umbilical cord blood of their neonates, as well as assessing the associations of these levels with the development of fetal macrosomia and diabetic fetopathy. Materials and methods: This prospective study included 88 women with a singleton pregnancy and cesarean delivery. All the patients provided informed consent for participation. The study groups comprised individuals with type 1 diabetes mellitus (n = 32) and non-impaired glucose tolerance (n = 56). The levels of interferons alfa 2 and gamma, monokine induced by interferon-gamma, interleukin-1α, -1β, -2, -3, -4, -5, -6, -7, -8, -9, -10, -13, -15, -16, -17, -18, interleukin-1 receptor antagonist, interleukin-2 receptor alpha subunits, p40 and p70 subunits of interleukin-12, monocyte chemotactic protein-1, -3, macrophage inflammatory protein-1α, -1β, leukemia inhibitory factor, macrophage ingibitory factor, nerve growth factor beta, stem cell factor, cutaneous T-cell-attracting chemokine, growth-regulated oncogene alpha, hepatocyte growth factor, stem cell growth factor beta, stromal cell-derived factor-1 alfa, interferon gamma inducible protein 10, platelet-derived growth factor-bb, vascular endothelial growth factor, basic fibroblast growth factor, eotaxin, chemokine ligand 5, and macrophage, granulocyte, and granulocyte/macrophage colony-stimulating factors, tumor necrosis factor alfa and beta, and tumor necrosis factor-related apoptosis-inducing ligand were measured with multiplex immunoassay in the blood serum of women at 36–40 weeks of gestation and in the umbilical cord blood serum of their neonates. Results: Cytokine levels varied widely in the both study groups. Women with type 1 diabetes mellitus had higher levels of interleukin-1β, -6, -17, basic fibroblast growth factor, macrophage inflammatory protein-1α, -1β, and lower levels of interleukin-1ra compared to the control group. Their neonates had higher levels of interleukin-6, -8, granulocyte colony-stimulating factor, macrophage inflammatory protein-1α, -1β. Higher levels of interleukin-1β, -6, -17, and basic fibroblast growth factor in the blood serum of the women were associated with time in range less than 70% and the development of diabetic fetopathy. Time in range was inversely associated with the interferon gamma / interleukin-5 (ρ = −0.531; p = 0.013), interferon gamma / interleukin-10 (ρ = −0.441; p = 0.045), interleukin-2 / -10 (ρ = −0.473; p = 0.030), interleukin-1β / -10 (ρ = −0.561; p = 0.008), interleukin-1β / -1ra (ρ = −0.635; p = 0.002), and interleukin-6 / -10 (ρ = −0.540; p = 0.012) ratios. Conclusions: Cytokine profile alterations were the most pronounced in pregnant women with non-target time in range values and in those with diabetic fetopathy development. Serum cytokine levels correlate with glycemic profile parameters but are considered non-specific markers of pregnancy complications.

We aimed to determine fetal liver perfusion in PGDM and GDM pregnancies and to assess the relation of ductus venosus (DV) shunt fraction with adverse pregnancy outcomes. We conducted a prospective longitudinal observational study including 188 pregnant women: group I—patients with pregestational DM (PGDM, n = 86), group II—patients with gestational DM (GDM, n = 44), group III—control (n = 58). The patients included in the study underwent ultrasound examination at 30+0–40+0 weeks of pregnancy. We evaluated volumetric blood flow adjusted to EFW (Q, ml/min/kg) for umbilical vein, DV, left and main portal vein. The relative risk was calculated for adverse pregnancy outcomes. In PGDM pregnancies, umbilical blood flow was redistributed to the fetal liver, increasing left portal and total liver volumetric blood flow (p < 0.001) compared with GDM and control groups. Pathological reduction in the DV shunt fraction (≤ 16.5

This article presents the results of scientific research into the development and implementation of personalised technologies for reproductive disorders. It presents data on new platforms for gamete evaluation, preimplantation genetic diagnosis, diagnosis and therapy of endometrial dysfunction, engineering of transgenic constructs for treatment of gynecological and hereditary diseases, creation of a database of genomic information in the field of reproductive health and obstetric pathology (biobank “Genofond”), application of artificial intelligence to assess placental cell interactions.

Background: Uterine fibroids are a common benign tumor that negatively affects reproductive function. The criteria for tumor evolution have not yet been determined. It is assumed that tumor growth and the number of growth points are associated with the transformation of blood circulation in the uterus and myomatous node. Recently, data have appeared on the prospects for three-dimensional ultrasound to be used with a specialized program that allows for describing the characteristics of blood supply throughout the entire tumor volume at the level of the microvasculature, which has low speed parameters, namely Virtual Organ Computer-aided Analysis (VOCAL). Aim: The aim of this study was to assess the characteristics of the blood supply to intramural and intramural-subserous myomatous nodes of various sizes using ultrasound and power Doppler in three-dimensional mode using specialized VOCAL software. Materials and methods: This study included 40 patients with uterine fibroids with a single myomatous node of intramural-subserous or intramural localization (International Federation of Gynecology and Obstetrics types 4–6). Four study groups were formed. Group I consisted of ten patients with uterine fibroids with a node diameter of up to 4.0 cm inclusive. Group II comprised ten patients with a node diameter from 4.0 to 5.0 cm inclusive. Group III included ten patients with a node diameter from 5.0 to 8.0 cm inclusive. And Group IV included ten patients with a node diameter of more than 8.0 cm. Ultrasound examination was carried out using a W10-RUS ultrasound scanner (Samsung Medison Co., Ltd., South Korea) on days 5–7 of the menstrual cycle. Results: We found that the most vascularized were myomatous nodes with a diameter of up to 4.0 cm. With an increase in the diameter of myomatous nodes, vascular resistance increased, which in turn indicates a decrease in vascularization. In addition, a decrease in resistance in the central vessels of the tumor compared to its peripheral vessels indicates the characteristic development of the vascular network of the myoma. This is characterized by the formation of new blood vessels mainly from the periphery of the tumor, which form a highly vascularized vascular capsule, while the center of the myomatous node is a hypovascular core. Conclusions: Evaluation of uterine vascularization and uterine fibroids using the specialized VOCAL program allows us to describe the features of blood supply and re-evaluate the mechanisms of tumor development. The use of power Doppler in 3D mode with the use of the specialized VOCAL program makes it possible to further study the vascularization of tumor tissue and establish “hemodynamic variants” of the intramural form of uterine myoma to search for the relationship between tumor growth and the transformation of the associated myometrium.

BACKGROUND: Endothelial dysfunction is the leading pathogenetic factor of preeclampsia. The function of the endothelium may be reflected in its ability to form microvesicles, which are generated by cells through the regulated shedding of the plasma membrane. AIM: The aim of this study was to evaluate the endothelial microvesicles count in peripheral blood of women with normal pregnancy and pregnancy complications such as gestational arterial hypertension and severe preeclampsia. MATERIALS AND METHODS: This study included 72 individuals, of whom there were healthy non-pregnant women (n = 21), women with normal pregnancy (n = 20), pregnant women with gestational arterial hypertension (n = 24), and pregnant women with severe preeclampsia (n = 7). To isolate microvesicles from peripheral blood, the differential centrifugation method was used. Microvesicles were treated with antibodies to vascular endothelial growth factor receptors (VEGFR1, VEGFR2), CD41a, CD34, and CD31 conjugated to fluorochromes. The absolute and relative count of microvesicles, as well as the fluorescence intensity, were analyzed using a BD FACSCanto II cytofluorimeter. RESULTS: In normal pregnancy, the count of microvesicles with the VEGFR1+, VEGFR2+, CD31+, and CD34+ phenotype was increased compared to non-pregnant women. In gestational arterial hypertension compared to normal pregnancy, no differences were found in the endothelial microvesicles count and endothelial marker expression. In severe preeclampsia, the total microvesicles count and endothelial cell derived microvesicles count in the peripheral blood plasma decreased in comparison with normal pregnancy and gestational arterial hypertension. While the expression of endothelial markers such as VEGFR1, VEGFR2, and CD34 in microvesicles membranes in severe preeclampsia increased compared to normal pregnancy and gestational arterial hypertension. CONCLUSIONS: An increase in the endothelial microvesicles count in normal pregnancy may be associated with an increase in the vascular bed area due to placenta formation. A decrease in the endothelial microvesicles count in severe preeclampsia is associated with damage to the endothelium and disruption of its function. Increased expression of endothelial cell receptors on microvesicles in severe preeclampsia may reflect compensatory reactions of the endothelium during the above damage.

BACKGROUND: The development of new systems for the cultivation of embryos of model objects and the creation of embryonic models based on stem cells allow for describing the events of early embryogenesis in more detail. However, these models have a number of limitations, endorsing only indirect extrapolation of the knowledge gained to events occurring in vivo. Despite certain limitations, including those related to some differences between embryos developing in vivo and in vitro, implantation conditions in vivo and in vitro, and limited time for culturing human embryos, the study of post-implantation development has great prospects. AIM: The aim of this study was to evaluate the viability and potential for the development of human embryos from the blastocyst stage during their prolonged coculture with the endometrium in culture media designed to support the growth to the blastocyst stage. MATERIALS AND METHODS: Embryos obtained through assisted reproductive technology were cultured from the blastocyst stage to day 19st of development in culture media designed to support the growth to the blastocyst stage, in the presence of the endometrium. On day 19st, embryos and endometrial fragments were assessed under an inverted microscope using Hoffman modulation contrast. RESULTS: During visual assessment under an inverted microscope on day 19st of development and days 13–14st of coculture with the endometrium, viable developing embryos without signs of degradation were recorded, both freely located in a mass of undifferentiated cells and in direct contact with the endometrium. CONCLUSIONS: The data obtained indicate the ability of the embryo to further develop in the presence of the endometrium from the blastocyst stage to day 19st in in culture media that supports the growth to the blastocyst stage. This may serve as an experimental model for both evaluating endometrial receptivity in vitro and studying intercellular communication during implantation.

Introduction. Uropathogenic Escherichia coli (UPEC) are the dominant bacterial pathogens of urinary tract infections (UTIs). UPEC belong to different phylogenetic groups and have many virulence factors, the study of which, in conjuction with the assessment of their relationship with clinical forms of UTI, is necessary for a better understanding of the pathogenesis of UTI and the development of new diagnostic algorithms. Aim: determination of the molecular genetic characteristics of uropathogenic Escherichia coli isolated from pregnant women with asymptomatic bacteriuria. Materials and methods. Clinical isolates of uropathogenic E. coli (n = 70) from pregnant women with asymptomatic bacteriuria were included in the study. The PCR method was used to determine the belonging to phylogenetic groups and detect 15 virulence markers — genes associated with adhesion (fimH, papC, sfa, afa, focG); toxin synthesis (cnf 1, hlyA, sat, vat, usp); siderophores (fyuA, iroN, iuc); capsular antigen (kpsMII). To assess the statistical significance of differences, Fisher's exact test was used. Differences were considered statistically significant at a confidence interval of 95% (p 0.05). Results. Most of the UPEC isolates belonged to phylogroup B2 (51,4%) and were characterized by the detection of all UPEC-associated virulence factors included in this study; genes associated with adhesion (sfa, focG), invasins (ibeA), synthesis of toxins (hlyA, cnf1, vat, usp) and capsule (kpsMII), siderophores (fyuA, iroN, hlyA) were detected significantly more frequently (p 0.05). Two or more virulence determinants were detected in 93% of isolates. Conclusion. The identification of key determinants of virulence and/or a combination of virulence genes can be a prognostic marker for predicting the course of UTI, especially in pregnant women, and will expand diagnostic capabilities taking into account the virulent properties of the uropathogen.

BACKGROUND: Today, preclinical diagnosis of preeclampsia presents significant difficulties. In widespread practice, it is diagnosed based on existing clinical signs and laboratory and functional research methods. Most of them are invasive and expensive, which makes it difficult to use them in widespread clinical practice for diagnosis and, especially, for monitoring the effectiveness of therapy over the dynamics of the disease. It is known that the pathogenesis of preeclampsia can have two independent development paths, converging in a common resulting link — the formation of endothelial dysfunction. Methods for studying endothelial function include determining markers of its imbalance in blood samples and non-invasive functional tests. Non-invasive diagnosis of endothelial dysfunction using the EndoPAT test allows us to quantify endothelium-mediated changes in vascular tone during 5-minute occlusion of the brachial artery. AIM: The aim of this study was to evaluate the method for determining the endothelium function in the first, second and third trimesters of pregnancy during ongoing pathogenetic prevention of preeclampsia. MATERIALS AND METHODS: This interventional uncontrolled study of the effectiveness of preventing preeclampsia using non-invasive assessment of vascular endothelial dysfunction during pregnancy was conducted at the Research Institute of Obstetrics, Gynecology and Reproductology named after D.O. Ott, St. Petersburg, Russia. The study involved 108 pregnant women at high risk of developing preeclampsia. All pregnant women underwent a cuff test to determine endothelial dysfunction using the peripheral arterial tonometry technique on the Endo-PAT 2000 device. The dynamic study was carried out in the first, second and third trimesters of pregnancy. As a result of the study, when endothelial dysfunction was detected [logarithmic transformation of reactive hyperemia peripheral arterial tonometry index (LnRHI) less 0.51], a complex glycosaminoglycan was additionally added to the basic prophylaxis with acetylsalicylic acid at a dosage of 250 MU (one capsule three times a day for eight weeks), then a cuff test was monitored after 6–12 weeks. Statistical analysis was performed using the IBM SPSS Statistics 20 software. All tests for significance were two-tailed, and differences were considered significant at p 0.05. RESULTS: Endothelial dysfunction was detected in the first trimester in 59 (55%) patients, then during ongoing complex therapy in the second trimester (n = 72) in 46 (64%) patients and in the third trimester (n = 46) in 4 (1%) patients. Moderate preeclampsia in the third trimester (35–39 weeks of gestation) developed in 28 (25.9%) patients out of 108. At the same time, at the start of the study, 15 patients with endothelial dysfunction received complex therapy, and 13 individuals only took acetylsalicylic acid. Among 46 patients who observed the dynamics of the entire pregnancy, only 4 (8.7%) women developed preeclampsia. After complex treatment prescribed based on the first trimester parameters, out of 36 patients who discontinued complex therapy and did not undergo a functional test subsequently, preeclampsia developed in 16 (44.4%) women. In the second trimester, out of 26 patients who stopped complex therapy, preeclampsia developed in 8 (30.8%) people. Thus, constant monitoring and complex therapy reduced the frequency of preeclampsia. In the group with a history of preeclampsia, the disease developed in 13 (39.4%) women. In the group with a high risk of preeclampsia according to the results of combined prenatal screening in the first trimester, with the exception of a history of preeclampsia (blood pressure test, placental growth factor level in the blood serum, lowest uterine artery pulsatility index value calculated to assess the individual risk of preeclampsia with a titer less than 1 : 100), preeclampsia occurred in 4 (13.3%) women. In the group with extragenital pathology associated with the risk of preeclampsia, including obesity, pregestational diabetes mellitus, chronic arterial hypertension, and chronic kidney disease, with the exception of a history of preeclampsia and a high risk of preeclampsia according to the results of perinatal screening, preeclampsia occurred in 11 (24.4%) women. In the high–risk groups for preeclampsia, we identified the highest-risk group, namely, one with the presence of preeclampsia in the anamnesis. CONCLUSIONS: The effectiveness of the functional method for determining endothelial dysfunction in the first, second and third trimesters of pregnancy during pathogenetic prevention of preeclampsia has been proven.

Genetic constructs must be delivered selectively to target tissues and intracellular compartments at the necessary concentrations in order to achieve the maximum therapeutic effect in gene therapy. Development of targeted carriers for non-viral delivery of nucleic acids into cells, including those in muscle, which is one of the most challenging tissues to transfect in vivo, remains a topical issue. We have studied ternary complexes of plasmid DNA and an arginine–histidine-rich peptide-based carrier coated with a glutamate–histidine-rich polymer bearing skeletal muscle targeting peptide (SMTP) for the gene delivery to muscle tissue. The relaxation of the ternary complexes after polyanion treatment was assessed using the ethidium bromide displacement assay. The developed polyplexes were used to transfect C2C12 myoblasts in full-media conditions, followed by analysis of their toxic properties using the Alamar Blue assay and expression analysis of lacZ and GFP reporter genes. After delivering plasmids containing the GFP and lacZ genes into the femoral muscles of mdx mice, which are model of Duchenne muscular dystrophy, GFP fluorescence and β-galactosidase activity were detected. We observed that the modification of ternary polyplexes with 10 mol% of SMTP ligand resulted in a 2.3-fold increase in lacZ gene expression when compared to unmodified control polyplexes in vivo. Thus, we have demonstrated that the developed DNA/carrier complexes and SMTP-modified coating are nontoxic, are stable against polyanion-induced relaxation, and can provide targeted gene delivery to muscle cells and tissues. The results of this study are useful for a range of therapeutic applications, from immunization to amelioration of inherited neuromuscular diseases.

Ganglioside-monosialic acid (GM1) gangliosidosis (ICD-10: E75.1; OMIM: 230500, 230600, 230650) is a rare autosomal recessive hereditary disease, lysosomal storage disorder caused by mutations in the GLB1 gene that lead to the absence or insufficiency of β-galactosidase. In this study, we report a case of a Russian family with a history of GM1 gangliosidosis. The family had a child who, from the age of 6 months, experienced a gradual loss of developmental skills, marked by muscle flaccidity, psychomotor retardation, hepatosplenomegaly, and the onset of tonic seizures by the age of 8 months. Funduscopic examination revealed a «cherry red spot» in the macula, which is crucial for the diagnosis of lipid storage disorders. To find the pathogenic variants responsible for these clinical symptoms, the next-generation sequencing approach was used. The analysis revealed two variants in the heterozygous state: a frameshift variant c.699delG (rs1452318343, ClinVar ID 928700) in exon 6 and a missense variant c.809A>C (rs371546950, ClinVar ID 198727) in exon 8 of the GLB1 gene. The spouses were advised to plan the pregnancy with assisted reproductive technology (ART), followed by preimplantation genetic testing for monogenic disorder (PGT-M) on the embryos. Trophectoderm biopsy was performed on 8 out of 10 resulting embryos at the blastocyst stage. To perform PGT-M, we developed a novel testing system, allowing for direct analysis of disease-causing mutations, as well as haplotype analysis based on the study of polymorphic markers—short tandem repeats (STR), located upstream and downstream of the GLB1 gene. The results showed that four embryos were heterozygous carriers of pathogenic variants in the GLB1 gene (#1, 2, 5, 8). Two embryos had a compound heterozygous genotype (#3, 4), while the embryos #7 and 9 did not carry disease-causing alleles of the GLB1 gene. The embryo #7 without pathogenic variants was transferred after consideration of its morphology and growth rate. Prenatal diagnosis in the first trimester showed the absence of the variants analyzed in the GLB1 gene in the fetus. The pregnancy resulted in the delivery of a female infant who did not inherit the disease-causing variants in the GLB1 gene.

Spinal muscular atrophy 5q (SMA) is one of the most common neuromuscular inherited diseases and is the most common genetic cause of infant mortality. SMA is associated with homozygous deletion of exon 7 in the SMN1 gene. Recently developed drugs can improve the motor functions of infants with SMA when they are treated in the pre-symptomatic stage. With aim of providing an early diagnosis, newborn screening (NBS) for SMA using a real-time PCR assay with dried blood spots (DBS) was performed from January 2022 through November 2022 in Saint Petersburg, which is a representative Russian megapolis. Here, 36,140 newborns were screened by the GenomeX real-time PCR-based screening test, and three genotypes were identified: homozygous deletion carriers (4 newborns), heterozygous carriers (772 newborns), and wild-type individuals (35,364 newborns). The disease status of all four newborns that screened positive for the homozygous SMN1 deletion was confirmed by alternate methods. Two of the newborns had two copies of SMN2, and two of the newborns had three copies. We determined the incidence of spinal muscular atrophy in Saint Petersburg to be 1 in 9035 and the SMA carrier frequency to be 1 in 47. In conclusion, providing timely information regarding SMN1, confirmation of disease status, and SMN2 copy number as part of the SMA newborn-screening algorithm can significantly improve clinical follow-up, testing of family members, and treatment of patients with SMA.