Extraembryonic Structures Research Articles

Impact of storage period on hatching egg quality, extra-embryonic structures, embryo morphometry, hatchability, and Rosa 1 chick quality

Storage duration affects the biological value of hatching eggs. The study evaluated the composition and characteristics of hatching eggs from Rosa 1 hens, the morphometric traits of embryos and chicks, and hatching results based on storage time. A total of 1200 hatching eggs were divided into three groups: NS (control, non-stored), S4 (stored up to day 4), and S7 (stored up to day 7). Storage conditions were 17°C and approx. 60% relative humidity. Each group’s egg quality was measured for 30 eggs. Weight loss during storage was monitored. Lysozyme activity and viscosity were analyzed from fresh eggs and incubated eggs on days 7 and 14, using thin and thick albumen or amniotic fluid. Embryo morphometry was measured on days 7, 10, 14, and 18, and post-hatching results and the quality of one-day-old chicks were assessed. Eggs stored for 7 days (S7) had higher (P<0.05) weight loss compared to the NS group. The proportions of eggshell and yolk increased (P<0.05) after storage, and the yolk index decreased. Vitelline membrane strength was lower (P<0.001) in the S4 and S7 groups than in the NS group. The lowest (P<0.05) pH levels of thin, thick albumen and yolk were observed in the NS group. Lysozyme activity increased (P<0.001) in the thin albumen of eggs in the NS and S4 groups and the thick albumen from the NS group than in others. On day 7 of incubation, group S7 showed higher (P<0.05) lysozyme activity in thick albumen, with no activity in amniotic fluid. By day 14, lysozyme activity increased (P<0.001) in group S7’s thick albumen and S4 and S7 groups’ amniotic fluid. Viscosity was highest (P<0.05) in fresh eggs’ thin and thick albumen from group S7, but on day 7, thick albumen viscosity was lowest. On day 14, amniotic fluid viscosity was higher (P=0.009) in group S4 than in NS. Seven-day-old embryos in group S7 had lower (P=0.004) weight than others. Eye diameter in S7 embryos was also smaller on days 7 and 18. On day 10, NS group embryos had shorter body diagonal lengths. Storage up to 7 days adversely affected hatching efficiency. Storage affected the navel area of one-day-old chicks, with the highest neck length in male chicks from group S4 and the lowest in females from group S4. It is concluded that egg storage at 17°C and 60% relative humidity could be limited to 4 days to maintain hatchability and qualitative features.

Features of the state of extraembryonic structures in miscarriage early terms of pregnancy

Background. Despite the modern achievements of medicine, miscarriage remains the main problem of obstetrics and gynecology science and practice. Purpose – to assess the features of the state of extraembryonic structures in case of miscarriage in the early stages of gestation. Materials and Methods. Under observation were 32 patients (main group) with a verified threat of pregnancy termination with bleeding in the embryonic period of gestation (up to 10 weeks) and 20 women (control group) with a physiological course of early pregnancy. The exclusion criteria from the study were patients with induced pregnancy, multiple pregnancies, anomalies of the development of female genital organs. All patients underwent a standard clinical and laboratory examination. During the ultrasound examination, both the state of the embryo and the state of extraembryonic structures were evaluated: the volume of the fertilized egg (V f/e), the average internal diameter of the yolk sac, the nature of vascularization of the corpus luteum, the state of hemodynamics in the myometrium. During transvaginal ultrasound, the chorionic volume (Vc) was calculated and its vascularization index (VI) was evaluated using a special three-dimensional VOCAL program. Results. Analysis of embryo biometrics in the studied groups did not reveal any significant difference. When assessing the state of extra-embryonic structures, we found that the volume of the fertilized egg and the average internal diameter of the yolk sac in women with miscarriage in the embryonic period of gestation was significantly lower. The volume of the corpus luteum of the ovary in the main group was equal to 3.0±0.3 cm3 in the control group – 5.3±0.61 cm3 , (p &lt; 0.05). Vascularization of the corpus luteum in patients with bleeding in early pregnancy was also reduced. In patients with threatened termination of pregnancy with bleeding, Vc was 32.5±2.08 cm3 in control 71.6±2.4 cm3 . The vascularization index of the chorion in pregnant women of the main group was equal to 9.5±1.2, while in women with a physiological course of gestation it was 19.1±1.7 (p&lt;0.05). When evaluating the hemodynamics in the myometrium, it was also established that the resistance index in the uterine and spiral arteries in the patients of the main group was significantly higher compared to the pregnant women of the control group. Conclusions. When there is a threat of pregnancy termination with bleeding in the embryonic period of intrauterine development of the child, significant changes occur in extraembryonic structures. A decrease in the volume of the fertilized egg, the average internal diameter of the yolk sac, blood flow disorders in the corpus luteum and myometrium, a decrease in vascularization of the chorion and its hypoplasia create a high risk of developing placental dysfunction and worsen the prognosis of pregnancy.

Pheasant hatchability and physicochemical features of egg and extra-embryonic structures depending on eggshell color

The study aimed to analyze the biological value of eggs and extra-embryonic structures affecting pheasant hatchability depending on the eggshell's color. Eggs (1,415) from 62-wk-old pheasants were used. The quality of fresh blue (BL), brown (BR), and green (G) eggs were analyzed. Incubation lasted for 25 d. Thick albumen (d 0, 1, 7, 14), amniotic fluid (d 14, 18), and the yolk (d 0-14) were collected. The pH, viscosity, lysozyme activity, crude protein (CP) content in albumen and amnion, pH, vitelline membrane strength, and fatty acids (FA) content in the yolk were performed. The lowest hatchability was in the BL group, and the highest was in the G group. BL group showed lower eggshell thickness and strength and higher egg weight. In thick albumen and amniotic fluid, the pH decreased with the incubation. In the yolk, there was an increasing trend (P = 0.015), with a decrease on d 18 (P < 0.001). The vitelline membrane strength decreased after 1 d of incubation, excluding BR eggs (P < 0.001). Thick albumen viscosity was higher on d 14 in the G group than in other dates and groups, the lowest in amniotic fluid, and slightly higher in BL and BR eggs. On d 18, amniotic fluid viscosity increased (P < 0.001). The lowest viscosity was indicated in BL eggs (P < 0.001). The lysozyme activity in thick albumen on d 14 was the highest (uniquely in BR and G groups), and the lowest values were found in amniotic fluid on d 14; after four d, the activity increased (P < 0.001). The CP content was higher in the BL group on d 14. In amnion, on d 14, the CP content was the lowest (<1%) and increased on d 18 (P < 0.001). There was a higher FA content (especially UFA) in the G group and a decrease in FA content after d 14 (P < 0.001). It was found that eggs with green eggshells have the highest biological value, and blue eggs are the least useful for incubation.

Changes in physicochemical parameters of duck eggs and extra-embryonic structures during incubation

Duckling embryogenesis should be deepened due to the hatching technology and its modification possibilities. Many changes occur in incubated eggs, which expose the embryo to hazards. The study aimed to analyse the physicochemical properties of eggshell, yolk, thick albumen (TA), and amniotic fluid (AF) of incubated hatching eggs from 52-week-old Cherry Valley ducks. The morphological features of 18 fresh eggs were analysed. Over 28 days, a total of 800 eggs underwent incubation. Eggshell surface temperature and egg weight loss were measured on days 1, 4, 7, 10, 14, 18, 21, and 25. Eggshell, TA, AF, and yolk were collected from eggs at incubation days 1–21 (every week). TA was collected on days 0, 1, and 7, while AF on days 7, 14, and 21. The analysis covered a range of physicochemical parameters. Eggshell thickness decreased with incubation, reaching its lowest point posthatch (P < 0.001). The highest pH for TA was recorded on day 1, while the lowest was on day 7 when comparing days 0, 1, and 7 (P < 0.001). TA pH was consistently higher than in AF (P < 0.001). However, the pH of TA was the highest on day 1 and the lowest on day 7 (P < 0.001). Yolk pH increased from days 1 to 21 (P < 0.001). There was also a noticeable in egg weight loss (0.34% daily) (P < 0.001). Vitelline membrane strength decreased from day 0 to day 1 (P < 0.001). Lysozyme activity in thick albumen on day 7 was higher than on days 0 and 1 (P < 0.001). Lysozyme activity in AF was higher on day 21 than days 7 and 14 (P < 0.001). TA viscosity was highest on day 0 and lowest on day 1, compared to other days (P < 0.001). AF viscosity and CP content exhibited an increase on day 21 as compared to days 7 and 14 (P < 0.001). The CP content in TA was notably higher on day 7 than on days 0 and 1 (P < 0.001). Polyunsaturated fatty acids declined, while monounsaturated and transfatty acids increased (P < 0.001). Viscosity and lysozyme activity increased on day 7 in TA and day 21 in AF. TA and the amniotic cavity appeared to facilitate the transfer of substances, particularly CP. Viscosity could be an indicator for optimising incubation conditions, as incorrect changes can affect embryo mortality. The results showed the different utilisation of nutrients, such as fatty acids. It could support research on the in-ovo administration of various substances.

Angiogenic imbalance as a starting point for the development of placental dysfunction and pregnancy complications in patients with pathology of the fetal environment

In recent years, scientific views on perinatal protection of fetus have shifted to earlier periods of pregnancy – to the I trimester, because from this stage the formation of the fetoplacental complex and laying of organs and tissues take place, which in most cases determines the further course of pregnancy. The objective: to clarify the role of the imbalance of angiogenic growth factors as one of the pathogenetic mechanisms in the development of placental dysfunction, and to conduct a postnatal macromorphometric study of placenta and umbilical cord in women with pathology of the fetal environment. Materials and methods. The object of the study were 80 pregnant women (the main group) with a singleton pregnancy, selected from the cohort of patients from the risk group for the development of preeclampsia and miscarriage. Among them, at 30–32 weeks of gestation oligohydramnios was diagnosed in 40 patients (the first group) and polyhydramnios – in 40 women (the second group). The control group included 30 somatically healthy pregnant women with a normal volume of amniotic fluid and a physiological course of pregnancy.The concentrations of placental growth factor (PlGF), vascular endothelial growth factor-1 (VEGF-1), and an anti-angiogenic marker of neovasculogenesis, soluble fms-like tyrosine kinase (sFlt-1), were performed at 12–16 and 28–32 weeks of pregnancy by enzyme immunoassay method. The study of the structural and functional features of the placenta and umbilical cord included sonographic and organometric methods. The variational statistical method of analysis of the obtained results was carried out using the «STATISTICA for Windows®-6.0» package.Results. In the women of the main group in the I trimester the ultrasound signs of pathology of the embryo and extraembryonic structures were significantly more often observed: low placement of the fertilized egg (33.8 %), its deformation (27.5 %), and partial detachment of the chorion (28.5 %). At 12–16 weeks of pregnancy, a pronounced vascular imbalance was established with a 2.5-fold decrease in the concentration of PlGF in the case of oligohydramnios compared to control data, a 1.8-fold decrease in the level of free VEGF-1 (in the case of oligohydramnios), and an increased level of antiangiogenic growth factors (sFlt-1).The postnatal examination of the placenta and umbilical cord showed that in the pregnant women of the main group a predominance of abnormal forms of placenta (16.3 %), a predominance of eccentric and marginal types of attachment of the umbilical cord (53.8 %), main and intermediate types of vascular branching (63.7 %), changes in the diameter of the umbilical cord (17.5 %), a combination of these features in a third of cases (31.3 %) were found. In the women of the main group, there was a decrease of the placental-fetal coefficient compared to the individuals of the control group, which was especially pronounced in pregnant women with oligohydramnios (up to 0.11±0.01 units compared to control data - 0.16±0.01 units).Conclusions. In the period of 12–16 weeks of pregnancy a vascular imbalance was established at the stage of placentation in patients who were diagnosed pathology of the perinatal environment at 30–32 weeks. In particular, by oligohydramnios there is a 2.5-fold decrease in the concentration of PlGF compared to the control data, a 1.8-fold decrease in the level of VEGF-1, and an increased antiangiogenic growth factor (sFlt-1), which became a prerequisite for the development of placental dysfunction.The results of the postnatal examination of the placenta and umbilical cord became an indirect reflection of placentation conditions. They demonstrated the predominance of abnormal forms, the predominance of the eccentric and marginal type of attachment of the umbilical cord, the main and intermediate type of vascular branching, an increased rate of umbilical cord pathology (Warton’s jelly deficiency, anomalies of the umbilical cord vessels) and a decreased placental-fetal ratio in women with amniotic fluid pathology, which was it is especially pronounced with oligohydramnios.

ULTRASOUND CHARACTERISTICS OF THE DEVELOPMENT OF THE FETOPLACENTARY COMPLEX IN PREMIER PREGNANCY

The article is devoted to the study of ultrasound changes in the fetoplacental complex in the early gestational period in pregnant womenwith miscarriage. Placental insufficiency (PI) is a syndrome caused by morpho-functional changes and is the result of a complex responseof the fetus and placenta to various pathological conditions of the maternal body [1]. It is based on disorders of compensatory-adaptivemechanisms of the fetoplacental complex (FPC) at the molecular, cellular and tissue levels. Ultrasound fetometry and placentometry are very important methods for diagnosing the state of the fetoplacental complex and can be used from the early term. Many researchers identify a number of prognostically unfavourable ultrasound markers: low chorion attachment, especially with the appearance of areas of detachment, discrepancy between the size of the foetus and gestational age, lack of clear visualization of the embryo, especially its heartbeat. In pregnant women with a history of miscarriage it is important to identify early signs of placental dysfunction from the firsttrimester onwards and to provide adequate correction for this condition. For this situation it is important to create appropriate conditions for the development and growth of the placenta and its adequate functioning. Significant fetal abnormalities occur as early as the first trimester and have a significant impact on the further process of ontogenesis.Aims and objectives of the study. To assess the normal parameters of embryonic and extraembryonic formation and to evaluate theparticular features of the formation and development of the fetoplacental system during pregnancy in women with a history of miscarriage.Materials and Methods. We erformed a comprehensive ultrasonographic examination of 25 somatically healthy women with aphysiological gestational process (control group) between 5 - 40 weeks’ gestation and 25 pregnant women with a history of miscarriage (study group). The SONOACE 8800 GAI MT with a 3.5 to 7.5 MHz convex transducer was used in the study. The data were statistically processed using standard methods of mathematical analysis with the use of Student’s and Fisher’s criteria, standard and specialized computer programs. Paired correlation indices were used to study the nature and degree of correlation between different parameters. The scientific research was positively evaluated by the Bioethics Commission of the Bukovinian State Medical University (Protocol No. 4 of 6.12.2021).Results and their discussion. In the transvaginal examination, uncomplicated pregnancy is characterized by the mandatoryvisualization of the embryo in the fetal cavity with a diameter of 14 mm or more, corresponding to 6 weeks' gestation. From the time of embryo identification in our study, we measured the coccygeal-parietal size (CPS) and compared its values with gestational age. It was found that in 22 (88.0%) cases the embryo size corresponded to the calculated gestational age and the mean inner diameter of the fetal egg. In severe clinical manifestations of threatened miscarriage the most sensitive echographic sign was a decrease in the volume of the foetus, which is a marker of further adverse pregnancy course and outcome. However, when there was an isolated increase in myometrial tone in the absence of clinical manifestations of threatened abortion, there was predominantly an isolated reduction in amniotic cavityvolume, easily corrected by the administration of conventional antispasmodic therapy.The ultrasonic criteria for alterations in pregnancy were developed, including delay in embryo CTR by 2 weeks or more on ultrasound examination up to 9 weeks' gestation; bradycardia at 90 bpm or less up to 8-12 weeks' gestation; chorionic detachment withretrochorial hematoma formation (over 25 ml); tachycardia at over 200 bpm with clinical manifestations of spontaneous miscarriage; marked progressive decrease of amniotic cavity volume; severe polyhydramnios with a thick echopositive suspension in the amniotic cavity. The risk of spontaneous miscarriage and formation of placental insufficiency increases with the simultaneous detection of 2 or more echographic markers.Conclusions. On the basis of the above, it is possible to conclude on the need to conduct an ultrasound examination with an assessment of the echographic parameters of the formation and development of the embryo and extraembryonic structures in the first trimester in pregnant women with a history of miscarriage and a risk of developing placental insufficiency in the history in order toidentify markers of a complicated course of gestation and the and the subsequent choice of rational tactics of pregnancy management.

Features of the formation of pathogenetic changes of the placental complex on the background of intra-uteronomy infection

Abstract. One of the main complications during early embryogenesis and placentation is placental insufficiency. Under the influence of various factors, namely infectious factors, which, depending on the biological specificity, negatively affect the germ cells of the parents, the development of the embryo, the formation of the trophoblast and the placenta, which in turn leads to a violation of the function of the placental complex and gestational complications during pregnancy. Due to the expansion of diagnostic possibilities for detecting placental function disorders, as well as in connection with the emergence of new data on the mechanisms of blood circulation regulation in the placenta to assess the functional state of the feto-placental complex at the stage of the formation of primary placental insufficiency against the background of inflammatory diseases of the female genital organs, we were determined the dopplerometric pattern of blood flow (ovarian artery, intervillous space) in women of the main and control groups. Goal. To determine the dopplerometric pattern of blood flow characteristics (ovarian artery, chorionic volume, blood flow study in different areas of the chorion) in pregnant women against the background of inflammatory diseases of the female genital organs in the early trimester of pregnancy. Materials and methods. The main group consisted of 37 women with signs of inflammatory diseases of the female genital organs, the control group consisted of 30 practically healthy women. The results. Obtaining new data on the mechanisms of blood circulation regulation in the ovarian artery and intervillous space at the stage of formation of the placental complex in physiological and complicated pregnancy will allow to supplement the question of the pathogenesis of the development and prevention of placental insufficiency. Conclusions. The study of the blood flow of extraembryonic structures in the early stages of gestation allows to diagnose structural changes at the stage of placenta formation. Violation of blood flow in the first trimester of pregnancy against the background of an infectious factor that penetrates into various vascular links of the mother-placenta-fetus system with the deduction of blood flow volume indicators demonstrates the regularity of the formation of disturbed hemodynamics at the stage of formation of the placental complex. These indicators can serve as a criterion for the selection of women of the risk group for the formation of primary placental insufficiency.

EVALUATION OF CERTAIN BIOCHEMICAL AND ANGIOGENIC MARKERS IN THE FIRST TRIMESTER OF PREGNANCY IN WOMEN WITH UMBILICAL CORD PATHOLOGY

At the stage of formation of fetoplacental complex, various factors of the external and internal environment create prerequisites for subsequent unfavorable course of pregnancy and childbirth. Their influence falls on the period of formation and differentiation of extraembryonic structures in the first trimester of pregnancy. That is why umbilical cord pathologies are considered to be one of the main causes of the development of fetal distress, low birth weight and post-hypoxic states of newborns.&#x0D; The purpose of the research: to analyze changes in the parameters of certain biochemical and angiogenic markers in patients with umbilical cord pathology.&#x0D; Research materials and methods. Medical documentation of 56 patients (the main group) with umbilical cord pathology and abnormalities of its insertion (27 pregnant women with a single umbilical artery and 29 women with velamentous insertion) was processed. The comparison group included 30 patients with normal structural components of the umbilical cord. The level of free ȕ-hCG, PAPP-A, free estriol, AFP, placental lactogen, certain growth factors PIGF and sFlt-1 was evaluated using the method of immunoenzymatic analysis. “Astraia Obstetrics” software (“Astraia software gmbH”, Germany) was used to determine the risk of preeclampsia, chromosomal abnormalities, fetal growth retardation, and premature birth.&#x0D; Research results and their discussion. An imbalance in the concentration of PAPP-A and β-hCG, when its low levels corresponded to high levels of β-hCG, was revealed in the main group. An increase in the average values of the level of PAPP-A MoM was established in almost half of the observations (42.9%), a significantly higher level of free ȕ-hCG was noted in 28.6 % of samples. The combined indicators of the concentration of PlGF in women with umbilical cord pathology were significantly lower (by 1.6 times) compared to the data of the comparison group; placental lactogen has a tendency to decrease. The combined values of the concentration of PlGF in women with umbilical cord pathology were significantly lower (by 1.6 times) compared to the data of the comparison group, and the indicators of the level of sFlt-1 – by 1.5 times (p&lt;0.05). This made it necessary to evaluate the calculated parameters of the angiogenic coefficient, which reflects the depth of vascular disorders and the severity of vascular imbalance.. At the same time, this ratio was found to be 2.0 times lower in the patients of the comparison group compared to the combined total indicator in the main group - in pregnant women with umbilical cord pathology - 155.7 units. (p&lt;0.05)&#x0D; Conclusions. Among pregnant women with umbilical cord pathology, a high proportion of the increased PAPP-A indicator, probably a lower level of free β-chorionic gonadotropin subunit, and low parameters of placental lactogen in the third trimester of pregnancy were observed. The combined indicators of the concentration of PlGF in women with umbilical cord pathology were significantly lower (by 1.6 times), and the indicators of sFlt-1 level – by 1.5 times higher compared to the data of the comparison group, which can be a marker of the abnormal formation of fetoplacental complex with the deterioration of the intrauterine fetal condition, as well as an increase in the angiogenic factor. The use of “Astraia Obstetrics” module in combination with prenatal biochemical screening made it possible to assign 23.2 % of examined patients with umbilical cord pathology to the risk group for the development of chromosomal abnormalities, preeclampsia, fetal growth retardation and premature birth.

Origin, form and function of extraembryonic structures in teleost fishes.

Teleost eggs have evolved a highly derived early developmental pattern within vertebrates as a result of the meroblastic cleavage pattern, giving rise to a polar stratified architecture containing a large acellular yolk and a small cellular blastoderm on top. Besides the acellular yolk, the teleost-specific yolk syncytial layer (YSL) and the superficial epithelial enveloping layer are recognized as extraembryonic structures that play critical roles throughout embryonic development. They provide enriched microenvironments in which molecular feedback loops, cellular interactions and mechanical signals emerge to sculpt, among other things, embryonic patterning along the dorsoventral and left-right axes, mesendodermal specification and the execution of morphogenetic movements in the early embryo and during organogenesis. An emerging concept points to a critical role of extraembryonic structures in reinforcing early genetic and morphogenetic programmes in reciprocal coordination with the embryonic blastoderm, providing the necessary boundary conditions for development to proceed. In addition, the role of the enveloping cell layer in providing mechanical, osmotic and immunological protection during early stages of development, and the autonomous nutritional support provided by the yolk and YSL, have probably been key aspects that have enabled the massive radiation of teleosts to colonize every ecological niche on the Earth. This article is part of the theme issue 'Extraembryonic tissues: exploring concepts, definitions and functions across the animal kingdom'.

Extraembryonic tissues: exploring concepts, definitions and functions across the animal kingdom.

Extraembryonic tissues: exploring concepts, definitions and functions across the animal kingdom.

Extraembryonic tissue in chelicerates: a review and outlook.

The formation of extraembryonic membranes (EEMs) contributes to the proper development of many animals. In arthropods, the formation and function of EEMs have been studied best in insects. Regarding the development of extraembryonic tissue in chelicerates (spiders and relatives), most information is available for spiders (Araneae). Especially two populations of cells have been considered to represent EEMs in spiders. The first of these potential EEMs develops shortly after egg deposition, opposite to a radially symmetrical germ disc that forms in one hemisphere of the egg and encloses the yolk. The second tissue, which has been described as being extraembryonic is the so-called dorsal field, which is required to cover the dorsal part of the developing spider germ rudiment before proper dorsal closure. In this review, we summarize the current knowledge regarding the formation of potential extraembryonic structures in the Chelicerata. We describe the early embryogenesis of spiders and other chelicerates, with a special focus on the formation of the potential extraembryonic tissues.This article is part of the theme issue ‘Extraembryonic tissues: exploring concepts, definitions and functions across the animal kingdom’.

Keratin filaments mediate the expansion of extra-embryonic membranes in the post-gastrulation mouse embryo.

Mesoderm arises at gastrulation and contributes to both the mouse embryo proper and its extra-embryonic membranes. Two-photon live imaging of embryos bearing a keratin reporter allowed recording filament nucleation and elongation in the extra-embryonic region. Upon separation of amniotic and exocoelomic cavities, keratin 8 formed apical cables co-aligned across multiple cells in the amnion, allantois, and blood islands. An influence of substrate rigidity and composition on cell behavior and keratin content was observed in mesoderm explants. Embryos lacking all keratin filaments displayed a deflated extra-embryonic cavity, a narrow thick amnion, and a short allantois. Single-cell RNA sequencing of sorted mesoderm cells and micro-dissected amnion, chorion, and allantois, provided an atlas of transcriptomes with germ layer and regional information. It defined the cytoskeleton and adhesion expression profile of mesoderm-derived keratin 8-enriched cells lining the exocoelomic cavity. Those findings indicate a novel role for keratin filaments in the expansion of extra-embryonic structures and suggest mechanisms of mesoderm adaptation to the environment.

ULTRASOUND CHARACTERISTIC OF EMBRYO, FETAL EGG AND CHORIONIC STRUCTURES IN PREGNANT WOMEN WITH MISCARRIAGE

The aim: The aim of the study was to assess the peculiarities of the formation and development of the fetoplacental system, to study the structures of the embryo, gestational sac, chorion in pregnant women with miscarriage. Materials and methods: A comprehensive ultrasound examination of 50 pregnant women was carried out in the period from 5 to 16 weeks of pregnancy, of which 25 - with a history of miscarriage (main group), and 25 - with an unremarkable medical history (control group). Results: We have identified the following echographic markers of adverse course and outcome of pregnancy in women with miscarriage in embryonic and early fetal periods:- lag of CRL of an embryo by 2 weeks and more at ultrasound examination in terms up to 9 weeks of gestation;- corporal or basal (near the stem of the embryo body) location of chorionic detachment with the formation of retrochorial hematoma with a volume of more than 25 ml;- pronounced progressive decrease in the volume of the gestational sac and amniotic cavity;- pronounced polyhydramnios with the presence of a coarse echopositive suspension in the amnioticcavity. The likelihood of spontaneous miscarriage and the formation of placental dysfunction is higher with the simultaneous detection of 2 or more echographic markers. Conclusions: Ultrasound examination is necessary to assess the echographic parameters of the formation and development of the embryo and extraembryonic structures in the first trimester with a history of miscarriage in order to the subsequent choice of rational tactics of pregnancy management.

Eggshell decalcification and skeletal mineralization during chicken embryonic development: defining candidate genes in the chorioallantoic membrane

During chicken embryonic development, skeleton calcification mainly relies on the eggshell, whose minerals are progressively solubilized and transported to the embryo via the chorioallantoic membrane (CAM). However, the molecular components involved in this process remain undefined. We assessed eggshell demineralization and calcification of the embryo skeleton after 12 and 16 d of incubation, and analyzed the expression of several candidate genes in the CAM: carbonic anhydrases that are likely involved in secretion of protons for eggshell dissolution (CA2, CA4, CA9), ions transporters and regulators (CALB1, SLC4A1, ATP6V1B2, SGK1, SCGN, PKD2) and vitamin-D binding protein (GC).Our results confirmed that eggshell weight, thickness, and strength decreased during incubation, with a concomitant increase in calcification of embryonic skeletal system. In the CAM, the expression of CA2 increased during incubation while CA4 and CA9 were expressed at similar levels at both stages. SCL4A1 and SCGN were expressed, but not differentially, between the two stages, while the expression of ATP6V1B2 and PKD2 genes decreased. The expression of SGK1 and TRPV6 increased over time, although the expression of the latter gene was barely detectable. In parallel, we analyzed the expression of these candidate genes in the yolk sac (YS), which mediates the transfer of yolk minerals to the embryo during the first half of incubation. In YS, CA2 expression increases during incubation, similar to the CAM, while the expression of the other candidate genes decreases. Moreover, CALB1 and GC genes were found to be expressed during incubation in the YS, in contrast to the CAM where no expression of either was detected.This study demonstrates that the regulation of genes involved in the mobilization of egg minerals during embryonic development is different between the YS and CAM extraembryonic structures. Identification of the full suite of molecular components involved in the transfer of eggshell calcium to the embryo via the CAM should help to better understand the role of this structure in bone mineralization.

Algorithm for the diagnosis and treatment of primary placental dysfunction in pregnant women of risk groups

Research objective: to evaluate the effectiveness of diagnosis and complex therapy of placental dysfunction (PD) in early stages of gestation.Materials and methods. We examined 110 pregnant women with risk for the primary PD. Exclusion criteria were multiple pregnancies, anomalies of fetal development and internal genital organs of the mother, pregnancy resulting from assisted reproductive technologies.The diagnosis of PD was established in 56 women (group I) based on the assessing the state of extraembryonic structures and blood flow in the spiral and uterine arteries at 12–13 weeks of gestation. The rest of the patients (54 women) entered the control group without PD (group II).Results. All pregnant women with verified PD at 12–13 weeks were prescribed complex drug treatment, which included micronized progesterone, venotonic Normoven, Magnicum, and Artihol. As a result of treatment, already at 22–24 weeks of gestation, the average value of blood flow (resistance index) in the uterine and spiral arteries did not differ significantly in the groups. There was no significant difference in fetometry and placentometry indices at 35–36 weeks of gestation. Complications of the gestational period in patients in the study groups were also almost the same.Conclusions. History of hormonal disorders and miscarriage, clinical signs of miscarriage in early gestation, abnormal embryo placement, extragenital pathology, and genital tract infections are risk factors for PD. Determination of the state of extraembryonic structures at 7–8 weeks of pregnancy (ovum volume, blood flow in the corpus luteum) and at 12–13 weeks (chorion volume, vascularization index, blood flow in the spiral and uterine arteries) can serve as a verification criterion for the PD development. Complex drug therapy, which began from the early stages of gestation (micronized progesterone, Normoven, Magnicum, Artihol) has established itself as an effective method of treating primary PD in pregnant of risk groups.

Trophoblastic volumetric bloodflow evaluation in women with trombophylia on 7–10th week of pregnancy

Spontaneous abortion occur in 15–20% of all detected pregnancies, and 45–70% of all spontaneous miscarriages take place up to 10 weeks. Thrombophilia is one of the main causes of miscarriage and its effect on the course of pregnancy remains poorly understood. In this regard, there is great interest in the problem of diagnosing thrombophilia and further prevention of thrombosis in obstetric practice.Purpose. Compare the parameters of volumetric blood flow in the chorion with different types of trophoblastic blood flow disturbance in pregnant women with thrombophilia in the first trimester for their quantitative verification.Materials and methods. The study included 129 pregnant women at the 7th to 10th week of gestation with a diagnosis of thrombophilia and a control group - pregnant women with a normal course of this pregnancy and a successful outcome of past pregnancies.The patients were divided into III clinical groups:I. Pregnant women with a normal course of this pregnancy and a successful outcome of past pregnancies (n = 33) – comparison group;II. Pregnant women with thrombophilia, in whom current pregnancy occurred on anticoagulant therapy (n = 28);III. Pregnant women with thrombophilia without taking anticoagulant drugs before registration of pregnancy (n = 68).Ultrasound was performed on Voluson E8 and S8 machines. All pregnant women underwent transabdominal and transvaginal ultrasound in B-mode, in color-doppler (CD) mode, and volumetric reconstruction of chorion was performed using the VOCAL program. In B-mode, the state of the embryo and extraembryonic structures were examined. The identification of trophoblastic vessels was carried out using the CD – region of interest was placed in the trophoblast area detected in B-mode, size and shape was adapted for a particular section.Using three-dimensional echography, the chorion volume was performed. The degree of blood supply in chorionic volume were calculated; vascularization index (VI), flow index (FI) and perfusion index (VFI) were displayed on a histogram with quantitative indicators.ResultsIn the CD mode, the types of trophoblastic blood flow were identified in the chorionic vessels:1) continuous type – blood flow loci are continuously identified over the entire area of the basal surface of the trophoblast;2) intermittent type – blood flow loci are unevenly identified along the basal surface of the trophoblast;3) single loci – single blood flow loci are identified along the basal surface of the trophoblast;4) lack of blood flow – blood flow loci are not determined along the basal surface of the trophoblast.Assessment of the quantitative parameters of the volumetric blood flow was used in the corresponding type of blood flow to verify the classification of types of trophoblastic blood flow in the CD mode.Statistical analysis of the volumetric blood flow indices using three-dimensional chorionic reconstruction showed a high degree of reliability (p &lt; 0.0001) in the VI and VFI values between all corresponding types of blood flow.Discussion. High incidence of spontaneous abortion and the significant role of thrombophilia in this pathology leads to necessity of expanding the research in this area. There has been a lot of research done over the past decade. The quantitative threshold values of the resistance indices in the chorionic vessels were calculated using pulse-wave Doppler, the chorionic blood supply indices were determined using volumetric reconstruction for various pathologies.However, we have not previously met the classification of chorionic blood supply in the CD mode. This technique is very simple to perform and is a highly informative for predicting miscarriage in the first trimester. For the first time, we proposed the above classification of trophoblastic blood flow; for the first time, we proved the reliability of the classification developed by us using the method of volumetric reconstruction of the chorion with an assessment of trophoblastic blood flow, which allows us to recommend its use when writing a protocol for ultrasound of the fetus in the first trimester in pregnant women with thrombophilia.Conclusion. The classification of trophoblastic blood flow was verified by three-dimensional echography with quantitative indicators of the volumetric blood flow of the trophoblast vessels, with a high degree of reliability (p &lt; 0.001) which to indicates the reliability of this classification. It can be recommended to conduct a multicenter study for investigating fetal ultrasound with the additional use of the CD in the first trimester in pregnant women with a diagnosis of thrombophilia.

FUNCTIONAL CHANGES IN EXTRA-EMBRIONAL BLOOD FLOW DURING THE FORMATION OF THE PLACENTAL COMPLEX ON THE BACKGROUND OF INFLAMMATORY DISEASES OF THE FEMALE GENERAL ORGANS

The introduc tion of diagnostic ultrasonography into clinical medicine has given a powerful impetus to the development of practical obstetrics. The use of ultrasound methods of research makes it possible to widely use the possibilities of a comprehensive morphological and functional assessment of the embryo, chorion, extraembryonic structures, and not only to state the already existing pathology, but also in most cases to make judgments about the pathogenetic mechanisms that led to the development of pathological conditions. One of the main complications in early embryogenesis and placentation is placental dysfunction. Under the infl uence of various factors, namely infectious, depending on the biological specifi city, they negatively aff ect the germ cells of the parents, the development of the embryo, the formation of the trophoblast and placenta, which in turn leads to impaired function of the placental complex and gestational complications during pregnancy.Due to the expansion of diagnostic capabilities for detecting placental dysfunctions, as well as in connection with the emergence of new data on the mechanisms of regulation of blood circulation in the placenta, we determined the Dopplerometric picture of blood fl ow features (ovarian artery, intervillous space) in women of the main and control groups.According to the goal, an ultrasound examination was performed on 67 women at 6-7 and 12-14 weeks of gestation. The main group consisted of 37 women with signs of infl ammatory diseases of the female genital organs, the control group consisted of 30 women who were practically healthy.Obtaining new data on the mechanisms of regulation of blood circulation in the ovarian artery and intervillous space at the stage of formation of the placental complex during physiological and complicated pregnancy will complement the pathogenesis of the development and prevention of placental dysfunction.

Roles of cell differentiation factors in preimplantation development of domestic animals.

In mammalian embryos, the first visible differentiation event is the segregation of the inner cell mass (ICM) and trophectoderm (TE) during the transition from the morula to the blastocyst stage. The ICM, which is attached to the inside of the TE, develop into the fetus and extraembryonic tissues, while the TE, which is a single layer surrounding the fluid-filled cavity called the blastocoel, will provide extraembryonic structures such as the placenta. ICM/TE differentiation is regulated by the interaction between various transcriptional factors. However, little information is available on the segregation of the ICM and TE lineages in preimplantation embryos of domestic animals, such as cattle and pigs. This review focuses on the roles of cell differentiation factors that regulate the ICM/TE segregation of preimplantation bovine and porcine embryos. Understanding the mechanism of cell differentiation in early embryos is necessary to improve the in vitro production systems for bovine and porcine embryos.

Prenatal Diagnosis of Fetal Trisomy 5 Mosaicism with Congenital Pulmonary Airway Malformation Type 3: A Case Report

Background: Trisomy mosaicism of chromosome 5 is uncommon with few cases described. Case report: A 41-year-old woman underwent ultrasound (US) at 16 weeks, which showed oligohydramnios and intrauterine growth restriction (IUGR). Amniocentesis discovered a karyotype of 47,XX,+5/46,XX. US at 19 weeks disclosed IUGR, enlargement of right side of heart, main pulmonary artery dilatation, and a suspected congenital pulmonary airway malformation (CPAM) in the inferior lobe of the left lung. Due to poor fetal prognosis, the parents opted for legal termination of pregnancy. At postmortem, a wide ventricular septal defect and CPAM type 3 were found. Cytogenetic analyses on fetal tissues detected mosaic trisomy 5 in skin, thymus, kidneys and CPAM. Placenta and fetal peripheral blood revealed normal female karyotype. Discussion/conclusion: These results suggest that if a fetus presents normal phenotypic features, mosaicism may be confined to extraembryonic structures, otherwise, in case of malformations, it may be carried by affected organs.

Особенности морфофункционального состояния маточно-плацентарной единицы и эмбриона мыши на ранних сроках экспериментального невынашивания беременности

Introduction. The majority of cases of habitual miscarriage are closely related to impaired maternal immune tolerance. Experimental mice models of abortion, where the critical time points are established to monitor pregnancy progression, are widely used to study the pathogenesis of early reproductive loss. However, the relationship between the embryonic development and extraembryonic structures with the immune microen-vironment is not well understood. The aim of the study is to compare the morphofunctional characteristics of the uteroplacental unit and embryo in early pregnancy in mice with experimental spontaneous, induced, and potentiated abortions. Materials and methods. Allogeneic physiological pregnancy was modeled using ♀CBA × ♂BALB/c mice mating. Spontaneous abortions were reproduced by mating ♀CBA × ♂DBA/2 mice. Induced abortions and potentiated abortions were triggered by intraperitoneal administration of the immunomodulator ß-heptylglycoside muramyl dipeptide (MDP) to pregnant females on gestation day (gd) 5 and gd 7 mated in following combinations ♀CBA × ♂BALB/c and ♀CBA × ♂DBA/2, respectively. Mice were withdrawn from the experiment on gd 8, then uterine horns and uteroplacental units were isolated. The material was fixed in 10% neutral formalin, and standard processing was performed for histological examination. In equatorial sections, the cross-sectional area of uteroplacental unit was measured, and maternal, embryonic, as well as extraembryonic structures were analyzed. An immunohistochemical assay with rabbit polyclonal anti-CD68 antibodies was conducted to detect macrophages, then the density of CD68+ cells per 1 mm2 of the decidual area was determined. Results. On day 8 of pregnancy, in the groups of induced and potentiated abortions cross-sectional area of the uteroplacental units was decreased compared with the groups of physiological pregnancy and sponta-neous abortions. Induced and potentiated abortions demonstrated vasodilation in the mesometric decidua zone and stagnation of blood in the blood vessels, compaction of trophoblast, and hemorrhage in decidua basalis whether spontaneous abortions showed occluded maternal vessels surrounded by edematous and vacuolated trophoblast cells.At the early stages of modeling of spontaneous and immuno-dependent abortions, disturbances in the devel-opment of the embryo and extraembryonic structures were revealed, ranging from initial signs of resorption to the destruction of the embryo and membranes.In viable embryos, density indices of CD68+ macrophages in the decidual membrane of the uteroplacental units did not differ significantly in all miscarriage cases, CD68+ cells were not detected in cases of embryo-nal resorption. Conclusion. The MDP treatment in a period after completion of implantation and before forming a mature placenta (gd 5–7) results in the inducing or the potentiating abortions in CBA females mated with by Balb/c or DBA/2 males, respectively. In both cases, we revealed disturbed chorion morphogenesis and embryonic development to the extent of its complete resorption and the absence of CD68+ macrophages in resorbed embryos’ microenvironment on day 8 of pregnancy. Keywords: miscarriage, models, muramyl dipeptide β-heptylglycoside, uteroplacental unit, macrophages, immunotolerance