Fetal Intrauterine Development Research Articles

Epigenetic Genome Modifications during Pregnancy: The Impact of Essential Nutritional Supplements on DNA Methylation.

Pregnancy is an extremely stressful period in a pregnant woman's life. Currently, women's awareness of the proper course of pregnancy and its possible complications is constantly growing. Therefore, a significant percentage of women increasingly reach for various dietary supplements during gestation. Some of the most popular substances included in multi-ingredient supplements are folic acid and choline. Those substances are associated with positive effects on fetal intrauterine development and fewer possible pregnancy-associated complications. Recently, more and more attention has been paid to the impacts of specific environmental factors, such as diet, stress, physical activity, etc., on epigenetic modifications, understood as changes occurring in gene expression without the direct alteration of DNA sequences. Substances such as folic acid and choline may participate in epigenetic modifications by acting via a one-carbon cycle, leading to the methyl-group donor formation. Those nutrients may indirectly impact genome phenotype by influencing the process of DNA methylation. This review article presents the current state of knowledge on the use of folic acid and choline supplementation during pregnancy, taking into account their impacts on the maternal-fetal unit and possible pregnancy outcomes, and determining possible mechanisms of action, with particular emphasis on their possible impacts on epigenetic modifications.

Neurodevelopmental Manifestations and Birth Defects in Prader-Willi Syndrome: Findings from a Study in Western Ukraine

Prader-Willi syndrome (PWS) is a genetic disease characterized by mental retardation, as well as autism spectrum disorders, obesity and mood disorders. The aim of our study was to research a spectrum of genetic heterogenity and different clinical neurodevelopmental manifestations. Materials and methods: clinical, molecular, genetic and psychological methods were used. We observed 9 children (5 boys and 4 girls) with Prader-Willi syndrome from Lviv and Ivano-Frankivsk regions (Western Ukraine) who were diagnosed and followed-up at the Institute of Hereditary Pathology, National Academy of Medical Sciences of Ukraine, Lviv, for 4 years (2019–2022) and underwent molecular genetic testing. The ages of children ranged from 1 month to 7 years, with the mean value amounting to 3.6 years of age. Results: Our study revealed that 6 (66.7%) out of 9 children with PWS had birth defects. At ultrasound during pregnancy, we observed certain clinical features of the fetus, including fetal intrauterine development delay –5 (55.5%), and higher nuchal translucency measurement – 3 (33.3%). According to the checklist of autism spectrum disorders (CASD), 3 of the examined children with autism syndrome were at risk and required further monitoring, while 5 children scored 18-20 points, which indicated the presence of autistic spectrum and required in-depth monitoring during diagnosis. Stereotypical and repetitive play as a manifestation of problems with social interaction was observed in 3 boys with PWS. In our study, parents had no complaints about aggressive behavior in children with this syndrome. Conclusions: In cases if the children have signs of autism or developmental delay, they should undergo genetic counseling.

Morphological Changes in the After Birth Tissues in Intrauterine Chlamydial Infection

Bovine chlamydia causes great economic damage to agriculture due to the fact that the pathogen is widespread in the population, is extremely virulent and pathogenic for fetal intrauterine development, while in adults the disease has little symptomatic or latent course

Associations of maternal dietary patterns during pregnancy and fetal intrauterine development.

Dietary pattern is excellent in reflecting an individual's eating conditions. Longitudinal data on fetal growth can reflect the process of intrauterine growth. We aimed to evaluate the associations between maternal dietary patterns and intrauterine parameters in middle and late pregnancy. The present study was conducted within Jiangsu Birth Cohort (JBC) study. Dietary information was assessed with a food frequency questionnaire (FFQ) in the second and third trimester of gestation. B-ultrasound scans were performed to obtain fetal intrauterine parameters, including head circumference (HC), femur length (FL), abdominal circumference (AC), and estimated fetal weight (EFW). Exploratory factor analysis was used to extract dietary patterns. Multiple linear regression and linear mixed-effects model (LMM) were used to investigate the association between maternal dietary patterns and fetal growth. A total of 1,936 pregnant women were eligible for the study. We observed inverse associations of maternal “Vegetables and fish” and “Snack and less eggs” patterns during mid-pregnancy with fetal HC Z-score, respectively (“Vegetables and fish”: β = −0.09, 95% CI −0.12, −0.06; “Snack and less eggs”: β = −0.05, 95% CI −0.08, −0.02). On the contrary, “Animal internal organs, thallophyte and shellfish” pattern in the second trimester was associated with increased HC Z-scores (β = 0.04, 95% CI 0.02, 0.06). Consistently, score increase in “Vegetables and fish” pattern in the third trimester was inversely associated with the Z-scores of HC (β = −0.05, 95% CI −0.09, −0.02), while “Meat and less nuts” pattern was positively correlated with the Z-scores of HC (β = 0.04, 95% CI 0.02, 0.07). As compared to the fetus whose mothers at the lowest tertile of “Snack and less eggs” pattern in both trimesters, those whose mothers at the highest tertile demonstrated 1.08 fold (RR = 2.10, 95% CI 1.34–3.28) increased risk of small HC for gestational age (GA). No correlation was observed between maternal dietary patterns and other intrauterine parameters. Our results suggested the effects of maternal dietary patterns on fetal growth, particularly HC. These findings highlighted the adverse impact of unhealthy dietary pattern on fetal growth, might provide evidence for strategies to prevent intrauterine dysplasia and dietary guidelines during pregnancy.

Leucine supplementation during late gestation globally alters placental metabolism and nutrient transport via modulation of the PI3K/AKT/mTOR signaling pathway in sows.

In a previously published study we reported that sow dietary leucine supplementation during late pregnancy significantly improved newborn piglet birth weight by stimulating protein synthesis in the longissimus dorsi muscle. However, there is still limited knowledge as to whether leucine can exert its effects on the placenta, one of the most important temporal organs during pregnancy, to promote maternal-fetal nutrient supply and thus contribute to fetal intrauterine development. Therefore, we tested this hypothesis in the present study. In total, 150 sows at day 90 of gestation were divided into three groups and fed with either a control diet (CON), CON + 0.4% Leu or CON + 0.8% Leu, respectively, until parturition. Placental metabolomics, full spectrum amino acids and nutrient transporters were systematically analyzed after sample collection. The results indicated that Leu supplementation led to an altered placental metabolism with an increased number of metabolites related to glycolysis and the oxidation of fatty acids, as well as elevated levels of amino acid accumulation in the placenta. In addition, nutrient transporters of amino acids, glucose and fatty acids in the placenta were globally up-regulated and several enzymes related to energy metabolism, including hexokinase, succinate dehydrogenase, lactated hydrogenase, glycogen phosphorylase and hydroxyacyl-CoA-dehydrogenase, were also significantly increased with no change observed in the antioxidative status of those groups with Leu supplementation. Furthermore, the phosphorylation of PI3K, Akt, and mTOR was enhanced in the placenta of sows undergoing Leu treatment. Collectively, we concluded that supplementing the diets of sows with Leu during late gestation globally altered placental metabolism and promoted maternal-fetus nutrient transport (amino acids, glucose, and fatty acids) via modulation of the PI3K/Akt/mTOR signaling pathway.

Morphometric features of the fetal thyroid gland: ultrasound examination findings

The article discusses features of morphometric parameters of the thyroid gland based on results of ultrasound examination. The need to study development of the thyroid gland in the fetus is due to the high prevalence of thyroid pathology and the possibility of its early prenatal diagnostics.The aim of the study was to characterize morphometric parameters of the fetal thyroid gland at 20–24 weeks of gestation using ultrasound examination. Material and methods. In the study the thyroid glands of 70 fetuses, both male and female, at the age of 20-24 weeks, were exposed to ultrasound examination. During ultrasound examination, the authors evaluated linear parameters, an organ perimeter, thyroid volume, and growth rate. Results. Ultrasound examination of fetuses aged 20–24 weeks demonstrated that all anatomical structures of the thyroid gland were well visualized on the images. The period from the 20th to the 24th week of the fetal intrauterine development was accompanied by an increase in the studied parameters of the thyroid gland, the growth intensity ranged from 3% (width of the left lobe) to 37% (thyroid volume). In female fetuses, the lobe sizes of the thyroid gland and the index of the thyroid volume prevailed. This period was also characterized by a low (up to 14%) growth intensity of certain linear parameters in male and female fetuses in different age subgroups.Conclusion. Starting from the 20th week of intrauterine development, ultrasound examination allows detecting features of the topography, quantitative anatomy and growth intensity of the fetal thyroid gland. The data obtained evidence the possibility of assessing the state of the thyroid gland during ultrasound screening of a fetus.

Malnutrition Related Diabetes Mellitus in Indian Population

Malnutrition related diabetes mellitus (MRDM) is rare type of diabetes associated with long term malnutrition and is not uncommon in India. Diabetes mellitus is the most common endocrine disorder of multifactorial aetiology and has been linked to both chronic under nutrition and obesity. Malnutrition is often related to deficient micro and macro nutrients, unhealthy behaviours and low socioeconomic status. Various clinical studies have underlined the relationship between malnutrition and diabetes mellitus. In the present review, we mainly explored malnutrition related diabetes mellitus in Indian population. Maternal under nutrition during pregnancy causes a decrease in the fetal intrauterine development rate known as intrauterine growth retardation (IUGR). Malnutrition majorly causes persistent insulin deficiency, glucose intolerance and insulin resistance and there by increases the risk of diabetes in Indian population. In this study we have also discussed the pathogenesis of MRDM and their management like self- dietary management and evidence based individualized nutritional therapy.

Relationships between placental adiponectin, leptin, visfatin and resistin and birthweight in cattle.

Adipokines can affect intrauterine development while calf birthweight (CBW) is a breeding standard of calves, which reflects the status of fetal intrauterine development. To explore the correlation between placental adipokines and CBW, 54 healthy Chinese Holstein cows were used in the present study. The cows were grouped according to the CBW of their calves. Placentas were collected immediately after delivery and enzyme-linked immunosorbent assay and reverse transcription-polymerase chain reaction were used to detect the placental expression levels of adiponectin, leptin, visfatin and resistin. Our results show that the mRNA transcription and blood placental content of adiponectin, leptin, visfatin and resistin increased with increasing CBW. The analysis showed that the mRNA transcription levels of placental adiponectin, leptin and resistin were positively correlated with CBW. The mRNA and protein expression levels of adiponectin, leptin and visfatin between the three groups were significantly correlated. Placental resistin mRNA levels correlated positively with adiponectin mRNA, but not leptin or visfatin. The protein expression levels of resistin were significantly positively correlated with those of adiponectin, leptin and visfatin. These results suggest that placental adipokines play important roles in regulating calf intrauterine growth.

AB159. Endocrine disrupting chemicals: toxicological risk assessment in vivo and in vitro models

In several studies, scientists asserted that many of endocrine disruptors (EDs), which have been involved in developmental, reproductive, neural, immunological, and other problems in wildlife and laboratory animals. Some environmental EDs, such as di-(2-ethylhexyl) phthalate (DEHP), flutamide (Flu), parabens, are used in many products in life and environment. However, the adverse effects caused by EDs can be temporary or permanent and the mechanism(s) through which these chemicals elicit their effects on biological systems of human and animal health is not clearly understood. The specific aim of this study is to evaluate endocrine disrupting chemicals-induced impact on the male or female reproductive system. An attempt is also made to elucidate the impact of these EDs in an in vitro model, i.e., GH3 rat pituitary cell line. A great deal of work has been carried out on the toxicity of phthalate, Flu, parabens in vivo and in vitro models. In brief, studies have been indicated that long-term and short-term exposure to various endocrine disrupting compounds (i.e., DEHP, Flu, parabens) during development stage (i.e., gestation, neonatal, immature, peripubertal) were done to find alternative dysfunctions later in animal life. The development and function of male or female reproductive tract showed many abnormalities, e.g., menstrual cycle irregularities; impaired fertility, endometriosis, and polycystic ovarian syndrome in female or morphological and functional gonadal dysfunction, e.g., infertility and decreased libido, congenital malformations (altered embryonic and fetal intrauterine development) and testicular dysgenesis syndrome in male. In addition, the differential gene expression patterns by microarray analysis following EDs exposure were found, particularly in steroid hormone synthesis, androgen and/or estrogen synthesis, and sex determination-related gene. On the other hand, studies revealed that parabens, a weak estrogenic chemical, exerted their actions on the stimulation of CaBP-9k gene, an estrogenic biomarker, via binding to estrogen receptor and/or progesterone receptor in immature female rat and GH3 cell line. An increasing number of chemical compounds in the environment have been identified as endocrine disruptor in vivo and in vitro bioassay. A future challenge is required to confirm a theoretical toxicology and risk assessment of EDs for human and animal health.

159 EFFECTS OF EMBRYO TRANSFER IN A LARGER BREED ON POSTNATAL GROWTH AND GLUCOSE METABOLISM IN HORSES

In equids, the size of the uterus determines fetal intrauterine development, which in turn affects postnatal insulin sensitivity and growth rate. We induced intrauterine growth enhancement through embryo transfer using Pony (P), Saddlebred (S), and Draft (D) horses and studied growth and insulin sensitivity in foals from birth to one year of age. Control pregnancies of S-in-S (n = 14) and P-in-P (n = 10) were obtained by AI. Enhanced fetal growth was obtained by transferring S (S-in-D, n = 7) and P embryos (P-in-D, n = 5) into D mares. From birth to weaning (180 days), each foal was kept with its surrogate P, S, or D dam. At 3, 140, and 380 days, glucose clearance and pancreatic beta cell response to exogenous glucose were assessed with an intravenous glucose tolerance test (IVGTT). At 200 days, the euglycemic-hyperinsulinemic clamp method was used to determine the sensitivity and responsiveness of tissues to exogenous insulin. Plasma T3, T4, and IGF1 were assayed at 3 and 180 days. Data were analysed using one-way ANOVA and Tukey post hoc tests. S-in-S were heavier and taller than P-in-P from birth to 380 days (P < 0.001). Before weaning, plasma concentrations of several hormones involved in growth were lower in S-in-S than P-in-P (at 3 and 180 days, respectively, T3: P = 0.08 and P = 0.02, T4: P < 0.001 and P = 0.06, IGF1: P = 0.04 and P < 0.001). No difference was found in glucose regulation between these groups. In contrast, post-weaning insulin resistance was observed in P-in-P at 200 days (P < 0.001) and confirmed at 380 days where they exhibited slower glucose clearance (P = 0.03) associated with higher fasting glucose (P < 0.001) than S-in-S. Fetal growth was not enhanced in S-in-D with no difference in height and weight at birth. Although S-in-D grew faster from 30 to 140 days, growth rates were not different from S-in-S after weaning, weaning coinciding with lower T3 (P < 0.001) in S-in-D than in S-in-S. Glucose regulation was not different between the two groups, but insulin remains to be assayed at 140 and 380 days. Fetal growth was enhanced in P-in-D: at birth, they were heavier (P = 0.01) and taller (P < 0.001) than P-in-P. Growth of P-in-D was faster until weaning. No more difference, however, was observed between P-in-D and P-in-P at 380 days. Plasma concentrations of T3 (P = 0.03) and those of T4 (P < 0.001) were lower at 3 days and T3 was still lower at 180 days (P < 0.001) in P-in-D compared with P-in-P. Moreover, P-in-D developed early insulin resistance: insulin secretion was higher in P-in-D compared with P-in-P (P = 0.002) after IVGTT at 3 days. At 200 days, however, P-in-D and P-in-P had the same sensitivity to insulin. There was no difference in glucose clearance rates at 380 days, but P-in-D had lower fasting glucose (P < 0.001) than P-in-P. Insulin assays at 140 and 380 days are pending. In conclusion, these data indicate that transfer of a small breed embryo into a large breed mare and subsequent suckling by the recipient mare can enhance fetal and postnatal growth and affect the foal’s glycaemia and sensitivity to insulin at birth and in subsequent months. Ongoing work includes analyses of milk samples and effects on general health.

Beneficial effects of dietary fibre supplementation of a high-fat diet on fetal development in rats

The objective of the present study was to investigate the effects of the addition of fibre and the antioxidant N-acetylcysteine (NAC) to fat-rich diets on fetal intrauterine development in rats. A total of eighty virgin female Sprague-Dawley rats were fed a control diet, a high-fat diet (HF), a high-fat and high-fibre diet (HFF) or a high-fat NAC diet until day 19·5 of gestation. Maternal HFF consumption resulted in a significantly higher mean fetal number and placental weight than in the other groups (P < 0·05). The HFF diet significantly abrogated HF-induced decreases in maternal serum and placental superoxide anion and hydroxyl radical scavenging capacities (P < 0·05); partially abrogated HF-induced increases in maternal serum and placental malondialdehyde (MDA) and protein carbonyl concentrations (maternal serum MDA and placental protein carbonyl, P < 0·05); resulted in significantly higher fetal liver total superoxide dismutase (SOD), Cu- and Zn-containing SOD and Mn-containing SOD (Mn-SOD) activities than in the HF group (P < 0·05). Furthermore, mRNA expressions of hypoxia-inducible factor 1-α, thioredoxin 2 and Mn-SOD in fetal liver and Mn-SOD in fetal heart and placental GLUT3 in the HFF group were higher than those in the other groups (P < 0·05). The inclusion of dietary fibre in the HF diet was more effective than NAC supplementation in maintaining maternal serum and placental superoxide anion and hydroxyl radical scavenging capacities close to those of the control. These results suggest that maternal fibre intake during pregnancy is beneficial for fetal intrauterine development possibly through the improvement of maternal, placental and fetal antioxidant capacities and placental nutrient transfer capacity.

The beneficial effect of fiber supplementation in high- or low-fat diets on fetal development and antioxidant defense capacity in the rat

There is mounting evidence that an imbalance in oxidant/antioxidant activities plays a pivotal role in fetal development. To determine the effects of maternal intake of fat and fiber on fetal intrauterine development and antioxidant defense systems of rats. Virgin female Sprague-Dawley rats were randomly assigned to 4 groups according to diet: the low-fat, low-fiber group (LL); the low-fat, high-fiber group (LH); the high-fat, low-fiber group (HL); and the high-fat, high-fiber group (HH). The diets were fed 4weeks prior to breeding through day 17.5 of pregnancy. Dietary intakes of fiber (wheat bran and oat) and fat were quantitatively varied, while intakes of energy and essential nutrients were kept constant among the diets. Rats fed a fiber-rich diet had significantly improved fetal numbers, as well as enhanced activity of superoxide dismutase (SOD) and capacity of scavenging free radicals (p<0.05). Meanwhile, the placental malondialdehyde and protein carbonyl levels were affected by the diet fat and fiber levels (p<0.05). Compared with the LL group, the mRNA abundance of hypoxia-inducible factor 1α (HIF-1α) and thioredoxin-2 (Trx2) in the maternal liver and glutathione peroxidase 1 (GPx1) in the placenta and fetus were significantly downregulated in the HL group (p<0.05). Furthermore, rats fed a fiber-rich diet had significantly upregulated mRNA expressions of Cu,Zn-SOD, Mn-SOD, and HIF-1α in the maternal liver (p<0.05); Cu,Zn-SOD and Mn-SOD in the placenta (p<0.05); and Cu,Zn-SOD in the fetus (p<0.05). When energy intakes are equivalent, consumption of fiber in high- or low-fat diets benefits fetal development and growth, through improvements in maternal, placental, and fetal antioxidant defense capacities.

Absence of Blood and Lymphatic Vessels in the Developing Human Cornea

The normal human cornea is devoid of both blood and lymphatic vessels and actively maintains this avascularity (corneal angiogenic privilege). Whether and when corneal angiogenic privilege is achieved during development is unknown. This study analyzed whether the cornea is primarily devoid of both blood and lymphatic vessels during intrauterine development or whether secondary regression of pre-existing vessels occurs before delivery. Indirect double immunohistochemistry was performed on 4-microm serial pupil-optic disc sections of paraffin-embedded human eyes stillborn at gestational ages of 17 to 41 weeks with antibodies against von Willebrand factor (vWF; factor VIII-associated antigen) as a panendothelial marker and with antibodies against lymphatic vessel endothelial hyaluronate receptor 1 (LYVE1) as a marker specific for lymphatic vascular endothelium. Human corneas were devoid of both vWF+++/LYVE-1(-) blood vessels and vWF+/LYVE-1+++ lymphatic vessels at all time-points analyzed. In contrast, there were numerous blood and lymphatic vessels detectable in the adjacent conjunctiva. The normal human cornea is primarily avascular and devoid of both blood and lymphatic vessels. Corneal angiogenic privilege is already achieved very early during fetal intrauterine development. This suggests early and strong expression of both antiangiogenic and antilymphangiogenic factors in the human cornea during development.