Current views on the possibility of cervical insufficiency correction

The main role in spontaneous abortion in 2nd and 3d trimesters is assigned to cervical insufficiency. According to a number of researchers, bed rest, elevated lower limbs, restriction of physical activity, tocolysis, antibacterial therapy do not affect the prolongation of pregnancy and are ineffective for preventing premature spontaneous labor. Correction of cervical insufficiency can be carried out by a vaginal form of progesterone, cerclage, pessary. The use of vaginal progesterone is justified in women with recurrent miscarriage, a history of premature birth, and shortening of the cervix to less than 25 mm. Indications for surgical correction are limited to patients with habitual loss of pregnancy due to cervical weakness or a history of premature birth. In the absence of significant obstetric history, cerclage has no advantages over the use of progesterone. The optimal time for cerclage is up to 20 weeks of pregnancy. Unlike progesterone cerclage has complications, the frequency and severity of which are attributable to the timing and indications for correction. Transabdominal cerclage is performed only when there is a technical impossibility of vaginal access due to the absence of a vaginal part of the cervix or after unsuccessful attempts of vaginal cerclages. Most often, the use of a pessary is associated with the diagnosis of a short cervix in terms of more than 24 weeks of gestation in the absence of an aggravated history. The combined use of gestagens, pessary and cerclage does not increase the efficiency of carrying a singleton pregnancy. Methods for the prevention of preterm delivery in multiple pregnancy, such as the introduction of a specialized outpatient service, bed rest, antibacterial therapy, progesterone, preventive cerclage or the insertion of a pessary do not change the incidence and mortality of newborns

Противовирусная активность лекарственного препарата на основе РНК двуспиральной натриевой соли в отношении SARS-CoV-2 in vitro

Scientific relevance. Innate immune activation in the early phases of COVID-19 infection and subsequent interferon induction may help control viral replication and protect cells not yet infected with SARS-CoV-2. Thus, immunostimulants that induce interferon (IFN), including double-stranded RNA-based agents, are a promising means of post-exposure prophylaxis and treatment of COVID-19 at early stages.Aim. The study evaluated the in vitro antiviral activity of a double-stranded RNA sodium salt-based medicinal product against SARS-CoV-2.Materials and methods. The authors analysed the double-stranded RNA sodium salt-based medicinal product RADAMIN®VIRO using Vero cells and the Delta variant of SARS-CoV-2 (B.1.617). The virus titre was calculated as the tissue cytopathic dose that caused 50% cell death. The authors measured the content of IFN-α and IFN-γ in the culture fluid by enzyme immunoassay and assessed the viral load by real-time polymerase chain reaction (using the cycle threshold value) and by titration (using Vero cells).Results. The studied double-stranded RNA sodium salt-based medicinal product at a concentration of 250 or 500 μg/mL induced IFN-α and IFN-γ expression by Vero cells, thus increasing their resistance to SARS-CoV-2. The authors evaluated the antiviral activity of the medicinal product based on the virus titre, viral load, and cell monolayer damage. The antiviral activity became clear 24 h after treatment, which confirmed the ability of the medicinal product to inhibit the replication of the SARS-CoV-2 virus in vitro as early as the first day after infection.Conclusions. The double-stranded RNA sodium salt-based medicinal product induced IFN-α and IFN-γ synthesis in Vero cells, increasing their resistance to SARS-CoV-2 infection in vitro. These results demonstrate the immunomodulatory and antiviral potential of the medicinal product.Актуальность. Активация механизмов врожденного иммунитета на ранних фазах развития инфекции COVID-19 и, как следствие, последующая индукция продукции интерферонов может способствовать контролю репликации вируса и защите еще неинфицированных SARS-CoV-2 клеток. В связи с этим в качестве средств постконтактной профилактики и лечения COVID-19 на ранних этапах представляется перспективным применение иммуностимулирующих препаратов, вызывающих индукцию интерферонов, в том числе препаратов на основе двуспиральной РНК.Цель. Оценка противовирусной активности лекарственного препарата на основе РНК двуспиральной натриевой соли в отношении вируса SARS-CoV-2 in vitro.Материалы и методы. Препарат на основе РНК двуспиральной натриевой соли (РАДАМИН®ВИРО). Эксперименты выполняли на культуре клеток Vero. В исследовании использовали вариант дельта вируса SARS-CoV-2 (B.1.617). Проводили оценку цитопатического действия вируса. Титр вируса рассчитывали как показатель тканевой цитопатической дозы, вызывающей гибель 50% клеток. Содержание интерферонов α и γ в культуральной жидкости определяли с помощью метода иммуноферментного анализа, вирусную нагрузку – методом полимеразной цепной реакции в реальном времени (по показателю Ct) и титр вируса – титрованием на культуре клеток Vero.Результаты. Внесение препарата на основе РНК двуспиральной натриевой соли в концентрациях 250 мкг/мл и 500 мкг/мл к клеткам линии Vero приводит к индукции секреции интерферонов α и γ, что повышает резистентность клеток к заражению вирусом SARS-CoV-2. Противовирусная активность исследуемого препарата, оцениваемая по значениям показателей титра вируса, вирусной нагрузки и уровня поражения клеточного монослоя, отмечается через 24 ч после его воздействия, что показывает способность препарата задерживать размножение вируса SARS-CoV-2 in vitro уже в течение первых суток после заражения.Выводы. Препарат на основе РНК двуспиральной натриевой соли индуцирует синтез интерферонов α и γ клетками линии Vero, повышая устойчивость клеток к заражению SARS-CoV-2 in vitro, что свидетельствует о иммуномодулирующем и противовирусном потенциале исследованного препарата

NEONATAL MARFAN SYNDROME: CLINICAL DESCRIPTION AND COMPLEX APPROACH TO DIAGNOSTICS AND TREATMENT

Aim. Molecular-genetic tests for neonatal type of Mrfan syndrome make possible to clarify a dignosis in children with multiple phenotype anomalies and to choose correct treatment strategy.Material and methods. Medical-genetic testing and instrumental diagnostics (echo, Doppler, ECG, chest X-rays) made possible to guess the diagnosis of neonatal Marfan syndrome (MS). Direct DNA-diagnostics of MS for these patients including direct Senger-sequencing of the coding plots and neighbouring introne areas of exones 24-32 gene FBN1 completely proved the diagnosis. Results. First time in Russia in two non-relative families with newborns having multiple phenotype anomalies the diagnosis of MS was set at the first year of life and confirmed by molecular-genetic methods.Conclusion. The results of the study must be introduced into practice at specialized pediatric, cardiological and cardiosurgical centres and departments to estimate the risk of sudden death, choose treatment strategy, prescribe gene-specific therapy

DNA DIAGNOSTICS AND MUTATION SPECTRUM OF THE GENE FBN1 IN MARFAN’S SYNDROME

Aim. The development of an optimal protocol for diagnostic search for mutations with the use of the new generation sequencing technique (NGS) and evaluation of the mutation spectrum in Russian selection of the patients with Marfan syndrome.Material and methods. Totally 32 patients included with Marfan syndrome. For 24 the direct sequencing was done by Sanger, of 24-32 exons FBN1. For 10 persons the analysis performed of coding exons and close introns regiones of the gene FBN1 with the preparation of fragmented libraries and performing of NGS on the IonTorrent platform. For 12 persons the mutations search was done with the use of automatically developed panel of Ampliseq primers for multiplex amplification of coding regions of genes that are responsible for the connective tissue development.Results. In investigation of 24-32 exones of FBN1 we found 3 replacements (p.C921R, p.C950S and p.I1048T). In complete analysis of FBN1 gene by fragmentation check of replacements we found 4 premature stop-codons (p.Y181*, p.R516*, p.Q1811*, p.R2776*) and 3 missense variants (C739W, p.C1095S, p. C2468R). In addition, there was deletion with the shift of translation frame and occurence of stop-codon in the 9th exon (c.661delT). In one female patient there was replacement variant c.4942+4A>G, with non-defined clinical significance. With the complete analysis of FBN1 using Ampliseq there were 2 premature codons found (р. Q520*, p.K2838*) and 2 deletions with the translation frame shift (c.40_49del, c.6751del). In 6 from 12 there were missense replacements found (p.N2144S, p.A986T, p.C2390S, p.С2276W, p.C1777R and p.C2363G).Conclusion. In the case of absense of the “hot spot” exons, invention of NGS allows for optimization the search of mutations even in such long genes as the FBN1. Medical-genetic consultation and DNA-diagnostics are the integral methods for multidisciplinary care

REGULAR GENETIC COUNSELING AND DNA-DIAGNOSTICS OF MARFAN SYNDROME IN THE WORK OF FEDERAL SURGERY INSTITUTION

Aim. To invent a complex approach to patients with “marfanoid phenotype” undergoing surgery, applying the DNA-diagnostics of the gene FBN1 and medical genetic counseling.Material and methods. In the group of 37 patients with suspected Marfan syndrome we conducted analysis of coding exones and attached enthrones of the gene FBN1 with highly performing sequencing on platform IonTorrent.Results. After mutation screening in the sequences of gene FBN1, in 25 patients we confirmed the Marfan syndrome, and four of genetic mutation carriers did not have complete Ghent criteria. All genetic variants were analyzed and were applied at the stage of surgery planning for maximum radical result of surgical treatment and for medical genetic counseling of the families.Conclusion. The analysis performed, of clinical presentation, surgery indications and spectrum of post-operation complications in Marfan syndrome patients

PATHOGENETIC VALIDATION OF ADDITIONAL OBJECTIVE CRITERIA FOR POSSIBLE EFFECTIVE PREGNANCY PROLONGATION AFTER PREMATURE MEMBRANE RUPTURE

The goal of present study was a search for pathogenetical reasoning of an opportunity for prolongation of pregnancy complicated by premature rupture of membranes at a gestational term of 22-34 weeks. The patients were subject to due observation and expectant treatable of pregnancy with prevention of possible infectious and inflammatory complications, as well as monitoring of systemic inflammatory response markers, immune state, and cytokine profile of blood in pregnant women with this disorder. We conducted a comprehensive clinical and laboratory examination of fifty pregnant women, whose pregnancy was complicated by premature membrane rupture at 22-34 weeks of gestation. A control group consisted of 40 women with normal pregnancy. For assessment of cellular composition of the blood, a BC3000+ hematological analyzer was used. Distinct subsets of peripheral blood lymphocytes were studied by flow cytometry using monoclonal antibodies («FACS Calibur» «Becton Dickinson», USA). Blood levels of cytokines (IL-2, IL-6, IL-8, TNFα, IL-4, IL-10) were determined by ELISA using test systems (ZAO “VectorBest”, Novosibirsk, Russia). Stereoultrastructural study of membranes was performed with a scanning electron microscope «Hitachi S-450”. The findings suggest that the failure of membranes emerging du to systemic metabolic disorders and changes in peripheral blood cells (leukocytosis, lymphopenia due to CD19+ B lymphocytes). Moreover, one could observe reduced counts of CD16+CD56+ T cells (natural killer cells) that showed certain parallelism with increased levels of proinflammatory cytokines (IL-6, IL-8, TNFα) in blood from pregnant PROM, as well as a decrease in IL-10 and IL-4 contents antagonized their proinflammatory effects to certain extent. An opportunity of incomplete pregnancy prolongation for patients with premature rupture of membranes was based on thorough assessment of their somatic and obstetric status and general condition of the fetus, when adequate and comprehensive therapy was applied. In the course of pregnancy prolongation, we found a progressive increase in pro-inflammatory cytokine levels (IL-2, IL-6, IL-8, TNFα), a steady decrease in CD19+ B cell counts, CD3+СD4+ helpers, natural killer cells, increased levels of cytotoxic CD3+CD8+ T cells .The pathogenesis-based criteria for necessary termination of the pregnancy for women with PROM are identified, including an increase in acute-phase proteins levels in blood, development of neutrophilic leukocytosis, lymphopenia, increase of pro-inflammatory cytokine levels in blood (IL-1β, IL-6, IL-8, TNFα), along with progressive reduction of CD3+СD4+ lymphocytes, CD16+CD56+ and CD19+ В lymphocytes

Development of certified reference material (CRM) of uranium mass fraction in metallic uranium

Metallic uranium certified reference material (CRM) has been developed and certified as State certified reference material and Russian 1 class certified material (according to the RF State Standard 8609-2004) within the State system of accounting and control of nuclear materials. The certified parameter is mass fraction of uranium

<i>In vitro</i> antiviral activity of a double-stranded RNA sodium salt-based medicinal product against SARS-CoV-2

Scientific relevance. Innate immune activation in the early phases of COVID-19 infection and subsequent interferon induction may help control viral replication and protect cells not yet infected with SARS-CoV-2. Thus, immunostimulants that induce interferon (IFN), including double-stranded RNA-based agents, are a promising means of post-exposure prophylaxis and treatment of COVID-19 at early stages.Aim. The study evaluated the in vitro antiviral activity of a double-stranded RNA sodium salt-based medicinal product against SARS-CoV-2.Materials and methods. The authors analysed the double-stranded RNA sodium salt-based medicinal product RADAMIN®VIRO using Vero cells and the Delta variant of SARS-CoV-2 (B.1.617). The virus titre was calculated as the tissue cytopathic dose that caused 50% cell death. The authors measured the content of IFN-α and IFN-γ in the culture fluid by enzyme immunoassay and assessed the viral load by real-time polymerase chain reaction (using the cycle threshold value) and by titration (using Vero cells).Results. The studied double-stranded RNA sodium salt-based medicinal product at a concentration of 250 or 500 μg/mL induced IFN-α and IFN-γ expression by Vero cells, thus increasing their resistance to SARS-CoV-2. The authors evaluated the antiviral activity of the medicinal product based on the virus titre, viral load, and cell monolayer damage. The antiviral activity became clear 24 h after treatment, which confirmed the ability of the medicinal product to inhibit the replication of the SARS-CoV-2 virus in vitro as early as the first day after infection.Conclusions. The double-stranded RNA sodium salt-based medicinal product induced IFN-α and IFN-γ synthesis in Vero cells, increasing their resistance to SARS-CoV-2 infection in vitro. These results demonstrate the immunomodulatory and antiviral potential of the medicinal product