Peculiarities of proteome profile of eutopic endometrium in women with adenomyosis depending on fertility

Abstract In recent years, several studies have reported a correlation between adenomyosis and major obstetric adverse effects. Implantation failures can be caused by changes in the expression of some molecules, referred to as "implantation markers", which are expressed by the endometrium and are required for successful interaction between the embryo and endometrium. The purpose of the study was to determine the features of the proteomic profile of the endometrium in women with adenomyosis, depending on their fertility. Material and methods. A comparative immunohistochemical (IGC) analysis of 102 eutopic endometrial specimens from women of reproductive age with adenomyosis and infertility (group I) was performed; from 20 nonpregnant women with adenomyosis who had a history of desired pregnancy and ended in childbirth and did not record episodes of miscarriages (group II), as well as from 30 conditionally gynecologically healthy women who applied for the introduction of an intrauterine system for contraception (control group C). A biopsy of the eutopic endometrium was performed on P+6 day of the menstrual cycle, during the period of the expected implantation window, with the help of aspiration pipel-curette. In immunohistochemical (IGC) determination of the proteomic profile of the endometrium HOXA10 was stained with monoclonal antibodies (MAB) to HOXA10 (sc-17159, Santa Cruz Biotechnology, USA); LIF - murine MAB against LIF (J-14F: SC-80159, Santa Cruz Biotechnology Inc., USA); sgp130 - goat polyclonal antibodies (PAB) anti-gp130 (BAF 228, supplied 50 μg / ml, R&D Systems); IL-6 - primary rabbit antibodies (code No CXH1-066LS, supplied as rabbit IgG 20 µg / ml; Cambridge Bioscience, UK). Results. It was found that important factors of reproductive disorders in women with adenomyosis are: changes in the proteomic profile of the eutopic endometrium on the day of the expected implant window, such as a decrease in HOXA 10 expression in the stroma by 1.52 (p<0.01) and in the glands by 1.46 times (p<0.01), LIF in glands - in 1.12 (p<0.01) and sgp130 - in 2.52 (p<0.01), as well as increased secretion in glands of IL-6 in 1,99 (p<0.01) times. In fertile women with adenomyosis, less pronounced changes in the proteomic profile of the eutopic endometrium are observed, which creates the conditions for successful realization of reproductive intentions, including a decrease in expression of HOXA 10 in the stroma by 1.48 times (p0.05); secretion in glands of LIF - in 1.06 (p>0.05) and gp130 - in 1.98 (p<0.01); increased IL-6 1.08 times (p<0.05). Conclusions. Changes in the proteomic profile of the endometrium play an important role in the occurrence of reproductive disorders in adenomyosis

Complex treatment of vaginal dysbiosis in women with recurrent miscarriage in assisted reproductive treatment (ART)

The article presents literature data on modern approaches to diagnosis and treatment of vaginal dysbiosis caused by fungi of the genus Candida and personal data on the effectiveness of vaginal dysbiosis treatment in women with recurrent miscarriage occurring in ART cycles by combined local therapy with sertaconazole and povidone-iodine. 240 patients of reproductive age with habitual miscarriage of pregnancy after ART and 30 conditionally healthy fertile women were examined. The state of vaginal microbiota was studied using PCR and culture. It were carried out a species identification of the fungi of the genus Candida and their susceptibility to antimycotics. It has been established that among women with a recurrent miscarriage after cycles of ART that repeatedly receive antibacterial therapy in preparation in the ART cycles and after abortion, in the vaginal microbiota Vaginal dysbiosis was observed in 54.58% of the cases, in which increased colonization by bacterial-fungal associates was noted in 67.18% of individuals. The authors prove that Sertaconazole in combination with povidone iodine is a highly effective regimen of vaginal microbiota normalization in patients with ART with a recurrent miscarriage, even in individuals with the development of cross-resistance to antimycotic therapy

РОЛЬ ХАРАКТЕРИСТИК Я-КОНЦЕПЦІЇ У ЗУМОВЛЕННІ УСПІШНОСТІ ФУНКЦІОНУВАННЯ ЛЮДИНИ ЯК СУБ’ЄКТА ЖИТТЄДІЯЛЬНОСТІ

The results of empiric verification of hypothesis in relation to causal connection between the indexes of differentiation and clarity of I-conception of the personality and optimisticness of its prognoses on the future in the different sphere of vital functions are considered.Рассмотрены результаты эмпирической проверки гипотезы относительно каузаль­ной связи между показателями диференцированности и ясности Я-коицепцпи личности и оптимистичностью ее прогнозов на будущее в разной сфере жизнедеятельности.Розглянуто результати емпіричної перевірки гіпотези щодо каузального зв’язку між показниками диференційованості і ясності Я-концепції особистості та оптимістичністю її прогнозів на майбутнє у різних сферах життєдіяльності

ОСОБЛИВОСТІ ІМУННОЇ РЕАКТИВНОСТІ ЕНДОМЕТРІЯ З НЕАТИПОВОЮ ГІПЕРПЛАЗІЄЮ У ПАЦІЄНТОК ІЗ БЕЗПЛІДДЯМ ТА ОЖИРІННЯМ

The aim of the study – to learn the features of the expression of NK cell subpopulations in the endometrium with non-atypical hyperplasia in patients with infertility and obesity. Materials and Methods. The study included 84 infertile women with non-atypical endometrial hyperplasia (NAEH), including 44 with normal body weight and 40 with obesity, and 30 relatively healthy women in the control group. Endometrial samples were taken on the 22nd day of the menstrual cycle during the proposed implantation window in women with NAEH – with hysteroscopy, and in women of the control group – with pipel-biopsy. Immunohistochemical determination of CD16+- and CD56+ NK cells in the endometrium was carried out by the avidin-streptavidin-peroxidase method using the corresponding monoclonal antibodies. Results and Discussion. In infertile women with NAEH, there was an excessive activation of immune reactivity in the endometrium with increased infiltration of its CD56+ and cytotoxic CD16+ NK cells: the expression of CD56 + was 8.14 times higher than the same control parameters ((56.91±2.43) versus (7.18±0.41) ‰, p&lt;0.01), СD16+ – 48.04 times ((41.31±1.84) versus (0.88±0.15) ‰, p&lt;0.01). At the same time, the content of CD56 + NK cells in the endometrium of women with NAEH and obesity was 1.28 times lower than that in patients with normal body weight ((49.61±3.19) versus (63.55±3.34) ‰, p&lt;0.01); CD16 + content – 1.32 times ((35.32±3.14) versus (46.75±1.71) ‰, p&lt;0.01). Conclusions. Obesity in infertile women with NHE due to systemic chronic inflammation leads to a decrease in the number and cytotoxicity of NK cells in the endometrium during the expected window of implantation compared to a similar pathology in persons with normal body weight, which may be a factor in the progression of the corresponding neoplastic changes.Цель исследования – изучить особенности экспрессии субпопуляций NK-клеток в эндометрии с неатипической гиперплазией у пациенток с бесплодием и ожирением. Материалы и методы. Под наблюдением находились 84 бесплодных женщины с неатипической гиперплазией эндометрия (НГЭ), среди которых 44 с нормальной массой тела и 40 – с ожирением, и 30 условно здоровых женщин контрольной группы. Забор образцов эндометрия проводили на 22-й день менструального цикла во время предполагаемого окна имплантации у женщин с НГЭ – при гистероскопии, а у женщин контрольной группы – при пайпель-биопсии. Проводили иммуногистохимическое определение в эндометрии CD16+ и CD56+ NK-клеток авидин-стрептавидин-пероксидазным методом с использованием соответствующих моноклональных антител. Результаты исследования и их обсуждение. У бесплодных женщин с НГЭ имела место чрезмерная активизация иммунной реактивности в эндометрии с повышенной инфильтрацией его СD56+ и цитотоксическими СD16+ NK-клетками: экспрессия СD56+ была больше аналогичных показателей контроля в 8,14 раза ((56,91±2,43) против (7,18±0,41) ‰, p&lt;0,01), СD16+ – в 48,04 ((41,31±1,84) против (0,88±0,15) ‰, p&lt;0,01). В то же время содержание СD56+ NK-клеток в эндометрии женщин с НГЭ и ожирением было меньше по сравнению с аналогичным у пациенток с нормальной массой тела в 1,28 раза ((49,61±3,19) против (63,55±3,34) ‰, p&lt;0,01); содержание СD16+ – в 1,32 раза ((35,32±3,14) против (46,75±1,71) ‰, p&lt;0,01). Выводы. Ожирение у бесплодных женщин с НГЭ вследствие системного хронического воспаления приводит к уменьшению количества и цитотоксичности NK-клеток в эндометрии во время ожидаемого окна имплантации по сравнению с аналогичной патологией у лиц с нормальной массой тела, что может быть фактором прогрессии соответствующих неопластических изменений.Мета дослідження – вивчити особливості експресії субпопуляцій NK-клітин в ендометрії з неатиповою гіперплазією у пацієнток із безпліддям та ожирінням. Матеріали та методи. Під спостереженням перебували 84 безплідних жінки з неатиповою гіперплазією ендометрія (НГЕ), серед яких 44 з нормальною масою тіла і 40 – з ожирінням, та 30 умовно здорових жінок контрольної групи. Забір зразків ендометрія здійснювали на 22-й день менструального циклу під час очікуваного вікна імплантації у жінок із НГЕ – під час гістероскопії, а у жінок контрольної групи – під час пайпель-біопсії. Проводили імуногістохімічне визначення в ендометрії CD16+ і CD56+ NK-клітин авідин-стрептавідин-пероксидазним методом із використанням відповідних моноклональних антитіл. Результати дослідження та їх обговорення. У безплідних жінок із НГЕ мала місце надмірна активізація імунної реактивності в ендометрії з підвищеною інфільтрацією його СD56+ і цитотоксичними СD16+ NK-клітинами: експресія СD56+ була більшою від аналогічних показників контролю у 8,14 раза ((56,91±2,43) проти (7,18±0,41) ‰, p&lt;0,01), СD16+ NK-клітин – у 48,04 ((41,31±1,84) проти (0,88±0,15) ‰, p&lt;0,01). Водночас вміст СD56+ NK-клітин в ендометрії жінок із НГЕ та ожирінням був менший порівняно з аналогічним у пацієнток із нормальною масою тіла у 1,28 раза ((49,61±3,19) проти (63,55±3,34) ‰, p&lt;0,01); вміст СD16+ NK – у 1,32 раза ((35,32±3,14) проти (46,75±1,71) ‰, p&lt;0,01). Висновки. Ожиріння у безплідних жінок із НГЕ внаслідок системного хронічного запалення приводить до зменшення кількості та цитотоксичності NK-клітин в ендометрії під час очікуваного вікна імплантації порівняно з аналогічною патологією в осіб із нормальною масою тіла, що може бути фактором прогресії відповідних неопластичних змін

Лечение бесплодия у женщин с аденомиозом методом экстракорпорального оплодотворения (ЭКО)

У статті проведений аналіз репродуктивних результатів пацієнток гарних відповідачів з радіологічно вставленим діагнозом аденоміозу в циклах запліднення інвітро взагалі та в залежності від використання протоколів з агоністами та антагоністами ГнРГ. Автори роблять висновок про те, що у пацієнток з аденоміозом в циклах ЗІВ частота настання клінічних вагітностей порівняно з пацієнтками з трубним безпліддям менша; ранніх мимовільних абортів – вища, живонароджень – нижча. Протоколом вибору при проведенні запліднення інвітро у пацієнток з аденоміозом є довгий протокол з агоністами ГнРГ.The article analyzes the reproductive outcomes of patients good responderes with radiologically exposed diagnosis of adenomyosis in cycles of in vitro fertilization in general, and depending on the use of protocols with GnRH agonists and antagonists. The authors conclude that in patients with adenomyosis in cycles of in vitro fertilization, the frequency of clinical pregnancy compared with patients with tubal infertility is less; early spontaneous abortion is higher, live births is lower. Protocol of selection during IVF in patients with adenomyosis is a long protocol with GnRH agonists.В статье проведен анализ репродуктивных результатов пациенток хороших ответчиков с радиологично выставленным диагнозом аденомиоза в циклах оплодотворения in vitro в общем и в зависимости от использования протоколов с агонистами и антагонистами ГнРГ. Авторы делают вывод о том, что у пациенток с аденомиозом в циклах оплодотворения in vitro частота наступления клинических беременностей по сравнению с пациентками с трубным бесплодием меньше; ранних самопроизвольных абортов – выше, живорождений – ниже. Протоколом выбора при проведении ЭКО у пациенток с аденомиозом является длинный протокол с агонистами ГнРГ

Proliferative activity in ovarian endometrrioma

Endometriosis shows features similar to neoplasia, which recently increased interest in the monoclonal antigen Ki-67. The aim of the study was to study the expression of Ki-67 in the operated ovarian endometriomas in women of reproductive age. Materials of the study were 32 patients with ovarian endometriomas and 30 patients with tubal infertility in the control group who underwent surgical treatment. Immunohistochemical study was performed by avidin-biotin-peroxidase method according to the standard method. Evaluated the number of positively colored cells using rabbit monoclonal antibodies to Ki-67. In 68.75% of patients with operated endometriomas, active proliferation in ovarian formations is observed, which is manifested by increased expression of oncoprotein Ki-67 in the glandular epithelium, in the cytogenic stroma of the endometrium and in surrounding vessels. There is a direct correlation between the level of Ki-67 expression and the prevalence of the endometriomas

Molecular-genetic aspects of the endometrium state on the day of the tentative implantation window in women with recurrent miscarriage in the programs of assisted reproductive technologies

Golovatyuk K. P., Nosenko O. M., Makshayeva E. T., Trofimov D. Yu., Donnikov A. E., Kolin V. V. Molecular-genetic aspects of the endometrium state on the day of the tentative implantation window in women with recurrent miscarriage in the programs of assisted reproductive technologies. Journal of Education, Health and Sport. 2017;7(9):621-626. eISSN 2391-8306. DOI http://dx.doi.org/10.5281/zenodo.1068361 http://ojs.ukw.edu.pl/index.php/johs/article/view/5076 The journal has had 7 points in Ministry of Science and Higher Education parametric evaluation. Part B item 1223 (26.01.2017). 1223 Journal of Education, Health and Sport eISSN 2391-8306 7 © The Authors 2017; This article is published with open access at Licensee Open Journal Systems of Kazimierz Wielki University in Bydgoszcz, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. This is an open access article licensed under the terms of the Creative Commons Attribution Non Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 15.08.2017. Revised: 20.09.2017. Accepted: 29.09.2017. MOLECULAR-GENETIC ASPECTS OF THE ENDOMETRIUM STATE ON THE DAY OF THE tentative IMPLANTATION WINDOW IN WOMEN WITH recurrent miscarriage IN THE PROGRAMS OF ASSISTED REPRODUCTIVE TECHNOLOGIES 1K. P. Golovatyuk, 2O. M. Nosenko, 1E. T. Makshayeva, 3D. Yu. Trofimov, 4A. E. Donnikov, 3V. V. Kolin 1Limited Liability Company Medical Center for Reproductive Health "Gameta", Odessa, Ukraine; 2Odessa National Medical University, Ukraine; 3Limited Liability Company "NPF DNA-Technology", Moscow, Russia; 4FGBU Scientific Center for Obstetrics, Gynecology and Perinatology. named after acad. V.I. Kulakov Ministry of Health care and Social Development of Russia, Moscow, Russia; e-mail: [email protected] Abstract More than 50% of pregnant women after the programs of assisted reproductive technologies (ART) face the problem of recurrent miscarriage (RMC), especially in the first trimester. Significant role in the development of RMC has infectious factor and chronic inflammation in the endometrium. The aim: to reveal the peculiarities of immune response mRNA genes of the inflammatory component expression in the period of the tentative implantation window (TIW) in women with RMC in ART programs. Material and methods. The main group consisted of 240 patients with RMC in ART programs; the control group included 100 conditionally healthy fertile women. On the ground of PCR reverse transcription, the mRNA of the IL-1β, IL-2, IL-10, Foxp3, TLR9, IL-2Rα cytokine genes was examined in endometrial samples obtained with the help of biopsy on the TIW day. Results. Analysis of the transcriptional profile of the immune response genes in the endometrium on TIW day revealed that the relative level of mRNA expression of the IL-1β, IL-2, Foxp3, TLR9, IL-2Rα genes did not differ significantly in the main and control groups. Statistically significant decrease in mRNA expression of IL-10 gene was observed in women with RPL. Conclusions. A feature of mRNA expression of the inflammatory component of the immune response in TIW period in women with RMC in ART programs is a decrease in the expression level of the IL-10 gene mRNA, which may be one of the reasons for the unfavorable outcomes of the onset pregnancy. Key words: recurrent miscarriage, assisted reproductive technologies, immune response, reverse transcription-polymerase chain reaction, tentative implantation window

Immunometabolic changes in macrophages in response to house dust mite extract

To date, much remains unclear about the pathogenesis of asthma, one of the most common chronic and highly heterogenic diseases of the respiratory system. The lack of specific and highly effective therapy in case of certain asthma subtypes requires the search for new approaches to treatment. One possible approach would be to influence the metabolism and immune functions of myeloid cells. This approach finds its application in the treatment of cancer and other diseases in the pathogenesis of which macrophages play an important role. It was shown that the pathogenesis of allergic asthma in response to one of the most common allergens, house dust mite, is due to a metabolic TNF-mediated reprogramming of alveolar macrophages. This suggests that influencing the process of TNF production or metabolic adaptations with specific blockers may also lead to a reduction in the symptoms of the course of the disease as a whole. In this work, we experimentally tested whether the previously obtained phenotype that occurs in macrophages in response to HDM cultured in DMEM is preserved if cells are cultured under more physiologically relevant conditions: in a medium closely related in composition to blood plasma. We also analyzed open databases of alveolar macrophages sequencing obtained from patients with asthma or from the lungs of mice in an HDM-induced asthma model in order to correlate specific immunometabolic changes. It was found that macrophages cultured under conditions close to physiological, simultaneously increase the rates of respiration and glycolysis, and also produce TNF in response to HDM. The observed phenotype is consistent with transcriptomic analyzes performed on human and mouse samples, which revealed an increase in the expression of genes related to glycolysis, oxidative phosphorylation, and the TNF signaling pathway. Thus, the data confirm the relevance of the phenotype obtained in vitro to the changes occurring in the in vivo system. However, functional verification at the level of metabolites, proteins and changes in metabolic activity is also required. In addition, it remains to be established how the blocking of individual metabolic pathways affects the features of the functional macrophage phenotype that occurs in response to HDM, and whether this effect can alleviate asthma symptoms

Shear flow in a three-dimensional complex plasma in microgravity conditions

Shear flow in a three-dimensional complex plasma was experimentally studied in microgravity conditions using Plasmakristall-4 (PK-4) instrument on board the International Space Station (ISS). The shear flow was created in an extended suspension of microparticles by applying the radiation pressure force of the manipulation-laser beam. Individual particle trajectories in the flow were analyzed and from these, using the Navier-Stokes equation, an upper estimate of the complex plasma's kinematic viscosity was calculated in the range of $0.2$--$6.7~{\rm mm^2/s}$. This estimate is much lower than previously reported in ground-based experiments with 3D complex plasmas. Possible reasons of this difference are discussed.Comment: 5 pages, 4 figure