Introduction: Individual risk assessment of assisted reproductive technologies is essential for personalized treatment strategies. Genetic and genomic indicators of the response to stress by cells could provide individual prognostic indicators for in vitro fertilization (IVF) success. Such indicators include the copy number of ribosomal genes (rDNA), which modulates the level of protein synthesis, and the abundance of mitochondrial DNA (mtDNA), which provides the cell with energy, while the content of telomere repeats (TRs) indicate the biological age. Materials and Methods: The contents of the three repeats in DNA isolated from blood leukocytes of 40 women before and after ovarian stimulation were assayed prior to IVF. Then, we divided the women into a successful IVF group, IVF+ (N = 17, 7 cases of twins), and a group of failed cases, IVF– (N = 23). The control group included 17 non-pregnant women with natural childbirth in the past. The nonradioactive quantitative hybridization (NQH) method was applied to assay the genome repeat contents. Results: The number of rDNA copies in the IVF+ group was significantly higher than in the IVF– group (p < 10-8). The number of mtDNA copies in the IVF+ group also exceeded those in the IVF– group (p < 0.001), whereas the TR content in the two groups differed, albeit, non-significantly (p < 0.03). Following the ovarian stimulation, the rDNA copy numbers did not change, while the contents of the mtDNA and TR varied significantly. Conclusions: This pilot study has shown that rDNA abundance in blood leukocytes can be considered a stable and effective predictor. Very low numbers of ribosomal repeat copies (<330) entail a high risk of IVF failure. However, a combination of numerous mtDNA and TRs, provided that rDNA content is not very low, increases the probability of multiple pregnancies.

Prognozirovanie haraktera opuholej yaichnikov u beremennyh imeet vazhnoe klinicheskoe znachenie, no znachitel'no zatrudneno. V poslednie 5–10 let u neberemennyh v ginekologii predlozheno ispol'zovat' RMI (Risk of malignancy index), odnako v akusherskoj praktike cennost' etogo issledovaniya eshche ne ustanovlena. Cel'yu issledovaniya bylo opredelit' RMI-IV i RMI-V pri dooperacionnom neinvazivnom prognozirovanii haraktera opuholej yaichnikov u beremennyh. Retro i prospektivno otobrany dannye kliniko-laboratornogo obsledovaniya 114 beremennyh 20–38 let, iz kotoryh 15 pacientok imeli zlokachestvennye opuholi yaichnikov (ZOYA), 28 pacientok — pogranichnye opuholi yaichnikov (POYA) i 71 pacientka — dobrokachestvennye opuholi yaichnikov (DOYA). Provodili ul'trazvukovoe issledovanie (UZI) s ispol'zovaniem cvetovoj dopplerografii i impul'snovolnovoj dopplerometrii. Opredelyali koncentraciyu SA-125 s pomoshch'yu immunofermentnogo analiza. Dlya ocenki riska raka yaichnikov ispol'zovali modifikacii IV, V. V krovi beremennyh s ZOYA bylo vyyavleno umerennoe statisticheski ne znachimoe povyshenie SA-125 po sravneniyu s takovymi znacheniyami u pacientok s DOYA i POYA. Po sravneniyu s gruppoj beremennyh s DOYA, pacientki s POYA i ZOYA demonstrirovali povyshennyj uroven' RMI-IV i RMI-V. Dlya garantirovannogo razlichiya v diagnostike opuholej neobhodimy kraevye znacheniya (RMI-IV — vyshe 3500 ukazyvayut na ZOYA, nizhe 100 — na otsutstvie zlokachestvennogo processa). Dlya RMI-V analogichnymi znacheniyami yavlyayutsya 1500 i 60. Odnako dlya prinyatiya resheniya v bol'shinstve nablyudenij nalichiya tol'ko pokazatelej RMI-IV i RMI-V bylo nedostatochno i trebovalos' ispol'zovat' kompleksnyj podhod. Takim obrazom, opredelit' harakter novoobrazovanij yaichnikov u beremennyh trudno, esli ispol'zovat' tol'ko indeksy RMI. Dlya dooperacionnogo prognozirovaniya haraktera opuholej yaichnikov u beremennyh trebuetsya kompleksnoe klinicheskoe obsledovanie s ispol'zovaniem vizualizacionnyh metodov, primenenie modelej prognozirovaniya, uchityvayushchih bol'shoe kolichestvo opisatel'nyh «morfologicheskih» harakteristik opuholej.

Prediction of ovarian tumor type in pregnant women is of great clinical significance, however, it is vastly difficult. In the last 5–10 years gynecologists were suggested to use RMI (Risk of Malignancy Index) in non-pregnant women, however the value of the test for obstetric practice has yet to be established. The study was aimed to determine RMI-IV and RMI-V during preoperative non-invasive prediction of ovarian tumor type in pregnant women. Retrospective and prospective clinical and laboratory data of 114 pregnant women aged 20–38 were collected. Among them 15 patients had malignant ovarian tumors (MOTs), 28 had borderline ovarian tumors (BOTs), and 71 had benign ovarian tumors. Color Doppler and pulsed wave Doppler ultrasound was performed. The levels of СА-125 were defined by enzyme immunoassay. Models IV, V were used to assess the risk of ovarian cancer. A moderate non-significant increase in blood levels of СА-125 compared to patients with benign ovarian tumors and BOTs was found in pregnant women with MOTs. Patients with BOTs and MOTs showed higher RMI-IV and RMI-V values compared to the group of pregnant women with benign ovarian tumors. Extreme values are required to guarantee the differences in the diagnosis of tumors (RMI-IV > 3500 indicate the presence of MOTs, the values below 100 indicate no malignancy). Similar RMI-V values are 1500 and 60. However, in most cases, availability of RMI-IV and RMI-V is insufficient for decision making, and a comprehensive approach has to be used. Thus, it is difficult to define ovarian mass type in pregnant women using RMI only. Comprehensive clinical assessment with the use of imaging methods is required for preoperative prediction of ovarian mass type in pregnant women, along with the use of prognostic models taking into account the majority of descriptive “morphological” tumor characteristics.

Clinical observations of gastrointestinal (GIT) cancer in pregnant women are presented. It is emphasized that early warning signs, typical of the initial manifestations of gastrointestinal stromal tumors, remains underestimated against the background of pregnancy. Frequently, common stages of the disease (III–IV) are detected accidentally after the manifestation of symptoms (gastrointestinal bleeding, formations palpated through the abdominal wall) or due to the detection of metastatic ovarian tumors, which manifest themselves as an independent disease. Long-term results of treatment of patients with secondary ovarian malignancies remain unfavorable. Conclusion. Pregravid training should be at the forefront and include a complete examination of patients, especially those with untypical complaints, in level 3 institutions with appropriate laboratories and modern methods of instrumental research. Key words: pregnancy, gastrointestinal malignancies, ultrasound

Currently, surgical treatment aimed to exclude the malignant ovarian tumors is performed in almost 90% of patients with decidualized endometrial cysts (DEC). However, unnecessary surgical interventions increase the risk to maternal and fetal health. The study was aimed to perform a differential diagnosis of DEC in pregnant women in order to define the rational treatment. A total of 82 female patients were included in the study: 63 had endometrial cysts (EC), 16 had DEC, 3 had rare forms of endometriosis, and 10 had ovarian serous papillary borderline tumors. When performing the diagnostic ultrasound, our proposed model was used. The ultrasound imaging data obtained were juxtaposed with the concentration of the protein tumor markers (СА-125), the risk of malignancy index (RMI) was calculated, and the morphological assessment of the masses was performed. The ultrasound imaging parameters, being the most valuable for differential diagnosis of EC, DEC, and serous borderline tumors, were as follows: the altered mass wall thickness, the existence and shape of papillary masses, avascular echogenic inclusions with blurry contour, blood circulation and arrangement of blood vessels, ascites. The frequency analysis revealed the differences between groups based on the ultrasound imaging data (in 60–100% of observations). Histological examination revealed the differences between groups in 100% of observations. Our findings have made it impossible to prolong pregnancy in patients with DEC without performing surgery. The results of treatment provided to patients with DEC during pregnancy were worse compared to those in patients with no prominent decidualization in ovarian EC. Today, the diagnosis of DEC and the treatment of patients during pregnancy remain unsophisticated. Further clinical observation and the search for more reliable methods of the diagnosis and rational treatment of pregnant women with DEC are required.

The problem of ovarian tumor differential diagnosis is solved using comprehensive ultrasound examination at different levels of efficiency. However, ultrasound imaging is an operator-dependent technique due to subjective interpretation of results. The study was aimed to assess the ultrasound signs of benign and malignant ovarian germ cell tumors (OGCTs) in pregnant women in order to determine the feasibility of surgical treatment and chemotherapy. The study was carried out using the logistic regression models. A group of 199 pregnant women with benign and malignant OGCTs were enrolled. Of them, 183 patients had benign, and nine patients had malignant tumors. In order to assess menstrual function and fertility, seven pregnant women were enrolled, who had previously received treatment for malignant OGCTs. Pre-operative assessment results were compared with morphological assessment data. Organ-preserving surgical treatment was performed (unilateral adnexectomy); if necessary, the patients received cisplatin-based chemotherapy. Perinatal outcomes were assessed. The median observation time between the malignant OGCT detection and the end of the study was 66 months (12–240 months). It was found that comprehensive ultrasound examination and logistic regression models (sensitivity 100%, specificity 92.3%, overall accuracy 92.8%) enabled differential diagnosis of benign and malignant OGCTs. The number of unnecessary surgical procedures in patients with benign OGCTs was been limited, the pregnancy and childbirth outcomes were improved. Nine pregnant women received organ-preserving surgical treatment for malignant OGCTs, and three patients received chemotherapy after surgery, which allowed the patients to realize their reproductive potential.

Исследовано число копий рибосомного повтора в геномах беременных женщин с нормально протекающей и осложненной беременностью. Выявлено, что более половины образцов ДНК женщин с патологией беременности содержали либо больше, либо меньше копий рДНК в геномах по сравнению с контрольной группой. Таким образом, есть все основания полагать, что вариации числа копий рибосомного повтора могут быть параметрами, позволяющими выявить группы риска среди беременных женщин на ранних сроках гестации. Женщины с низким и высоким содержанием рДНК в геноме нуждаются в более строгом наблюдении в период беременности.

Problemu differencial'noj diagnostiki opuholej yaichnikov reshayut s pomoshch'yu kompleksnogo ul'trazvukovogo issledovaniya (UZI), nesmotrya na to chto ono yavlyaetsya operatorozavisimym metodom iz-za sub"ektivnoj interpretacii poluchennyh dannyh. Cel'yu issledovaniya bylo ocenit' UZ-priznaki dobrokachestvennyh i zlokachestvennyh germinogennyh opuholej yaichnikov (DGOYA i ZGOYA) u beremennyh dlya opredeleniya vozmozhnosti hirurgicheskogo lecheniya i provedeniya himioterapii (HT). Issledovanie provodili s pomoshch'yu logregressionnyh modelej. V nem uchastvovali 199 beremennyh zhenshchin s DGOYA i ZGOYA. Iz nih 183 pacientki imeli dobrokachestvennye opuholi, devyat' — zlokachestvennye. Dlya ocenki menstrual'noj funkcii i fertil'nosti v issledovanie byli vklyucheny sem' beremennyh posle provedennogo ranee lecheniya ZGOYA. Poluchennye na dooperacionnom etape rezul'taty obsledovaniya sopostavlyali s morfologicheskimi issledovaniyami. Vypolnyali hirurgicheskoe lechenie v organosohranyayushchem ob"eme (odnostoronnyuyu adneksektomiyu), pri neobhodimosti — HT na osnove cisplatina. Izuchali perinatal'nye iskhody. Mediana nablyudeniya s momenta vyyavleniya ZGOYA do okonchaniya issledovaniya sostavila 66 mesyacev (12–240 mesyacev). Obnaruzheno, chto differencirovat' DGOYA so ZGOYA vozmozhno s pomoshch'yu kompleksnogo UZI i logregressionnyh modelej (chuvstvitel'nost' metoda — 100%, specifichnost' — 92,3%, summarnaya tochnost' — 92,8%). Ogranicheno chislo necelesoobraznyh operacij pri DGOYA i uluchsheny iskhody beremennosti, rodov. Hirurgicheskoe lechenie ZGOYA u devyati beremennyh vypolneno v organosohranyayushchem ob"eme, u troih — s posleduyushchej HT, chto pozvolilo realizovat' reproduktivnyj potencial.

V nastoyashchee vremya pri nalichii decidualizirovannyh endometrioidnyh kist yaichnikov (DEK) hirurgicheskoe lechenie dlya isklyucheniya zlokachestvennyh opuholej yaichnikov vypolnyayut prakticheski v 90% sluchaev. Neopravdannye hirurgicheskie vmeshatel'stva pri etom uvelichivayut risk dlya zdorov'ya materi i ploda. Cel'yu issledovaniya bylo provesti differencial'nuyu diagnostiku DEK u beremennyh dlya opredeleniya racional'nogo lecheniya. V issledovanie vklyucheny 82 pacientki: 63 iz nih byli s endometrioidnymi kistami (EK), 16 — s DEK, 3 — s redkimi formami endometrioza i 10 — s seroznymi papillyarnymi pogranichnymi opuholyami yaichnikov. Dlya ul'trazvukovoj (UZ) diagnostiki ispol'zovali predlozhennuyu avtorami model'. Poluchennye UZ-dannye sopostavlyali s koncentraciej belka-onkomarkera (SA-125), rasschityvali indeks RMI (risk of malignancy index), vypolnyali morfologicheskoe issledovanie obrazovanij. Osobo cennymi UZ-parametrami pri differencial'noj diagnostike EK, DEK i pogranichnyh seroznyh opuholej byli: izmenennaya tolshchina stenki obrazovaniya, nalichie i forma papillyarnyh obrazovanij, avaskulyarnye ekhogennye vklyucheniya bez chetkih konturov, krovotok i raspolozhenie sosudov, ascit. Rezul'taty chastotnogo analiza pokazali razlichiya mezhdu gruppami po dannym UZI (v 60–100% nablyudenij). Gistologicheskoe issledovanie vyyavilo razlichie mezhdu gruppami v 100% nablyudenij. Poluchennye rezul'taty ne pozvolili nam prolongirovat' beremennost' pri DEK bez vypolneniya hirurgicheskogo lecheniya. Rezul'taty lecheniya pacientok s DEK vo vremya beremennosti byli huzhe takovyh bez vyrazhennoj decidual'noj transformacii EK yaichnikov. V nastoyashchee vremya diagnostika DEK i lechenie bol'nyh vo vremya beremennosti ostayutsya nesovershennymi. Neobhodimy dal'nejshie klinicheskie nablyudeniya i poisk bolee nadezhnyh sposobov diagnostiki i racional'nogo lecheniya beremennyh s DEK.

Borderline ovarian tumors (BOTs) are common in women in their reproductive years. In more than one-third of patients tumors are detected at the age of 15–29, the average age at initial diagnosis is 40. The study was aimed to improve methods for BOTs diagnosis in pregnancy and to determine the possibilities of organ preservation treatment. A group of 300 pregnant women with various tumor-like formations and ovarian tumors was examined. Of them, 25 patients had borderline epithelial tumors (22 patients had serous and 3 patients had mucinous tumors). Ultrasound examination together with blood serum CА-125, sFas, VEGF and IL6 level assessment were performed prior to surgery. The results obtained were compared with the results of morphological studies. Organ preservation and radical surgical treatment were carried out, and chemotherapy, if necessary. Perinatal outcomes were studied when performing the cross-comparison. It was discovered, that ultrasonography and logistic regression analysis made it possible to distinguish between benign ovarian tumors, BOTs and malignant ovarian tumors. The levels of VEGF above the 500 pg/ml, IL6 above the 8.1 pg/ml and CА-125 above the 300 U/ml indicated the high probability of malignant ovarian tumors in pregnant women. Only the morphological study of ovarian tissue, obtained regardless of surgical methods, ensured understanding of the ovarian tumor’s true nature during pregnancy. At the same time, in three pregnant women with ovarian tumors, the morphological examination revealed some tissue areas common both for BOTs and malignant ovarian tumors. Thus, the predominance of the tumor early stages, relatively mild course and, favorable prognosis in patients with BOTs make it possible to use gentle surgical treatment making it possible to preserve menstrual function and fertility.

Пограничные опухоли яичников характерны для женщин репродуктивного периода, более чем у трети больных опухоли выявляют в возрасте 15–29 лет, средний возраст при первичной постановке диагноза составляет 40 лет. Целью исследования было усовершенствовать методы диагностики пограничных опухолей яичников на фоне беременности и определить возможности выполнения органосохраняющего лечения. Обследовано 300 беременных с различными опухолевидными образованиями (ООЯ) и опухолями яичников (ОЯ), из которых 25 имели пограничные эпителиальные опухоли: 22 — серозные, три — муцинозные. До операции проводили УЗИ, определяли концентрацию в сыворотке крови СА-125, sFas, VEGF и IL6. Полученные результаты сопоставляли с морфологическими исследованиями. Проводили органосохраняющее и радикальное хирургическое лечение, при необходимости — химиотерапию. При перекрестном сравнении изучали перинатальные исходы. Обнаружено, что различить доброкачественные опухоли яичников от пограничных (ПОЯ) и злокачественных (ЗОЯ) возможно с помощью УЗИ и логрегрессионных моделей. Уровни VEGF выше 500 пг/мл, IL6 выше 8,1 пг/мл и СА-125 выше 300 ЕД/мл свидетельствуют о высокой вероятности ЗОЯ у беременных. И только морфологическое исследование тканей яичников, полученных независимо от хирургических способов, давало истинное представление о характере опухоли яичников у беременных. Вместе с тем у трех беременных с ОЯ при морфологическом исследовании выявлены участки ткани, характерные как для ПОЯ, так и для ЗОЯ. Таким образом, преобладание начальных форм опухолевого процесса, относительно благоприятное течение и прогноз при ПОЯ позволяют достаточно широко использовать хирургическое лечение щадящего характера с сохранением менструальной функции и фертильности.

Определено количество копий рибосомных повторов (рДНК) в геномах женщин с нормальной и осложненной беременностью, а также женщин, подвергшихся процедуре экстракорпорального оплодотворения (ЭКО). Кроме того, измеряли содержание GC-богатой рДНК в образцах внеклеточной ДНК (вкДНК), полученных от женщин с нормальной и осложненной беременностью. Показано, что геномы более половины женщин с патологией беременности содержали либо больше, либо меньше копий рДНК, чем у любой женщины из контрольной группы. Также обнаружено более высокое содержание рДНК во вкДНК пациенток с осложненной беременностью, что свидетельствует о наличии хронического процесса аномальной гибели клеток в группе женщин с патологией беременности. Можно сделать принципиальный вывод: поскольку беременность является тяжелой нагрузкой на организм женщины, для успешного вынашивания требуется сбалансированный биогенез рибосом. Женщины как с низкой, так и с очень высокой копийностью рДНК имеют более высокую вероятность повышенного уровня апоптоза и попадания в группу риска. Параметр «число копий рДНК в геноме женщины» может служить дополнительным прогностическим маркером потенциальных осложнений беременности у женщины. Женщины с низким или высоким количеством копий рибосомных генов в геноме нуждаются в более внимательном ведении беременности. Показатели количества копий рДНК в геномах женщин с неудачными попытками ЭКО были значимо ниже, чем в геномах двух остальных групп. Этот факт говорит о том, что копийность рДНК в геноме является одним из факторов, влияющих на успех процедуры ЭКО. Если индвивидуальное число копий рДНК в геноме женщины меньше, чем 330, высок риск неудачного ЭКО. Необходимы дальнейшие исследования данного вопроса. As pregnancy is a stressful load for a woman, any stress-resistance factor is relevant to it. According to recent reports, ribosomal gene copy number in the genome is associated with the individual stress-resistance. We determined copy numbers of ribosomal DNA (rDNA) in genomes of pregnant women with normal and complicated pregnancy, and women after in vitro fertilization (IVF) procedure. We also measured the contents of GC-rich rDNA in cell-free DNA (cfDNA) derived from normal controls and complicated pregnancy cases. We have shown that genomes of more than a half of DNA samples from women with pregnancy pathology harbor either more, or less rDNA copies than any woman from the control group. We also found higher rDNA contents in cfDNA isolated from complicated pregnancy cases suggesting the presence of a permanent cell death process in pathology cases. A principal conclusion can be made: women with low rDNA copy numbers and with very high numbers can have higher cell death rates and belong to the risk group. The parameter «rDNA copy number in woman’s genome» can be an additional prognostic marker for eventual pregnancy complications in the woman. The numbers of rDNA copies in the genomes of women with failed IVF attempts was significantly lower than in the genomes of patients with succesfull outcome, suggesting that rDNA copy number in the genome is one of the factors that affect the success of the IVF procedure. If the individual rDNA copy number is under 330, the risk of IVF failure is high. Further studies are warranted.

Placental pathology is accompanied by the activation of apoptosis in the trophoblast and the subsequent increase in the concentrations of microvesicles containing placental (or fetal) DNA accumulating in the maternal blood. Fragments of fetal DNA stimulate the release of nuclear and/or mitochondria) DNA fragments by neutrophils. Therefore, one can expect that complicated pregnancies will be characterized by the dramatic elevation of total cell-free DNA (cfDNA) levels in maternal plasma. The aim of this work was to study the dynamics of plasma cfDNA concentrations and the activity of DNase I, an enzyme involved in the elimination of cfDNA from the bloodstream, in nonpregnant and pregnant women. Our study recruited 40 healthy nonpregnant women, 40 women with uncomplicated pregnancies and 35 women with the intrauterine growth restriction (IUGR) of the fetus. We did not observe the elevation of the total cfDNA concentrations in the patients with complicated pregnancies. Moreover, cfDNA concentrations in their plasma were even lower (though this difference was statistically insignificant) than in healthy pregnant and nonpregnant women. The median values of cfDNA concentrations in the group of healthy nonpregnant women were 75.5 ng/ml; in the group of healthy pregnant women, 78.0 ng/ml; and in the patients with IUGR, 42.1 ng/ml. At the same time, we observed a significant increase in DNase I activity in the plasma of women with IUGR. The median DNase I activity in the groups of healthy pregnant and nonpregnant women was 3.0 and 3.4 IU/ml, respectively. In patients with different grades IUGR of the fetus this parameter was as high as 6.3 IU/m1 (p < 0.001). Increased DNase I activity in the plasma of women with complicated pregnancies indirectly suggests a transient elevation of circulating cfDNA levels. Our study shows that the high level of activity exhibited by the cfDNA elimination system impedes the analysis of cfDNA concentrations in complicated pregnancies and skews its results. However, if cfDNA, DNase I activity and the cfDNA/DNase I ratio were all taken into account, it could be possible to develop a tool for the monitoring of cell death in the mother throughout the entire pregnancy.

In this study, the proteomic approach based on high performance liquid chromatography connected with tandem mass spectrometry (LC-MS/MS) and bioinformatics analysis were applied to identify differentially abundant proteins in chorionic villus samples (CVS) from women with blighted ovum and normal pregnancy. We identified about 600 proteins in the solubilized fraction of CVS. Comparative proteomic analysis revealed differences in the content (Average Normalized Abundances) of 187 proteins in blighted ovum. These included 134 down-regulated proteins and 53 up-regulated proteins. According to bioinformatics analysis these proteins participate in a variety of metabolic processes, including alcohol and tricarboxylic acid metabolism, response to endoplasmic reticulum stress, small molecular catabolic process, cellular respiration, and others. Proteins that demonstrated growing content in blighted ovum were mainly encoded by genes located on chromosomes 7 and 16 whereas proteins which demonstrated reducing abundance were mainly encoded by genes located on chromosomes 1, 2, and 11. We also revealed changes in the content of proteins encoded by genes located on the human chromosome 18; they are involved in apoptotic and drug metabolic processes with an important role in early pregnancy loss. Our pilot results demonstrate the efficiency of the LC-MS/MS approach for detecting the differences at the qualitative and semi-quantitative levels in the protein profiles of the CVS at anembryonic pregnancy compared to normal gestation. We conclude that globally profiled and differentially regulated proteins of CVS are helpful in obtaining molecular insights into biological processes of the pregnancy pathology.

The sizes, compartments, wall thickness, contents quality, internal lining, presence of ovarian tissue, course of vessels and features of blood flow were studied in ovarian cysts/tumors in 197 pregnant women (12-39 weeks). Based on the clinical and ultrasound features with the help of statistical methods there were developed the most important criteria for the diagnosis of tumors and tumor-like formations of the ovaries during pregnancy.