Placental Biopsies Research Articles

A proper placental sampling for syncytin-1 analysis.

Syncytin-1 (gene ERVW-1) has been proposed as a marker of pre-eclampsia and malfunctions in placental development. Placenta is heterogeneous tissue, hence the method of biopsy can significantly affect the outcome of analyses. A total of 44 placentae were analyzed by taking 3-30 samples from each. Relative levels of ERVW-1 expression in the placental biopsies were characterized by RT-qPCR. Evaluation of ten biopsies from one placenta individually (not pooling them) is recommended due to the high variability of expression. No significant correlation was found between biopsy localization and level of ERVW-1 expression; therefore, random sampling is recommended. A long cut from the umbilical cord to the edge of the placenta is a convenient approach to placental sampling.

Gravidity-dependent associations between interferon response and birth weight in placental malaria

BackgroundMaternal malarial infection leads to poor perinatal outcomes, including low birth weight from preterm delivery and/or fetal growth restriction, particularly in primigravidas. In placental malaria, Plasmodium falciparum-infected red blood cells cause an inflammatory response that can interfere with maternal–fetal exchange, leading to poor growth. The type I interferon (IFN-I) pathway plays an immunomodulatory role in viral and bacterial infections, usually by suppressing inflammatory responses. However, its role in placental malaria is unknown. This study examines the cytokine responses in placental tissue from subsets of malaria-infected and uninfected women, and attempts to correlate them with particular birth outcomes.Methods40 whole placental biopsy samples were obtained from pregnant women at least 16 years of age recruited to a larger prospective chemoprevention trial against malaria. These were patients at Tororo District Hospital in Uganda, an area of high malaria endemicity where approximately 40% of women have evidence of malaria infection at delivery. They were regularly followed at a local clinic and monitored for fever, with blood smears performed then and at time of delivery to diagnose malaria infection. Placenta biopsies were taken for histological diagnosis of placental malaria, as well as quantitative PCR analysis of genes in the IFN-I pathway (IFN-β, IL-10 and MX-1). Parameters such as infant birth weight and gestational age were also recorded.ResultsHistological analysis revealed placental malaria in 18 samples, while 22 were found to be uninfected. RT-PCR analysis showed a four-fold increase in IFN-β and IL-10 expression in multigravidas with placental malaria when compared to gravidity-matched, uninfected controls. This effect was not observed in primigravidas. Interestingly, linear regression analysis showed a positive association between IFN-β levels and higher birth weights (β = 101.2 g per log2-fold increase in IFN-β expression, p = 0.042). This association was strongest in primigravidas with placental malaria (β = 339.0, p = 0.006).ConclusionsThese results demonstrate differential regulation of the IFN-I pathway in placental malaria according to gravidity, with the greatest anti-inflammatory response seen in multigravidas. The association between IFN-β levels and higher birth weight also suggests a protective role for IFN-I against fetal growth restriction in placental malaria.

Monochorionic twins with selective fetal growth restriction: insight from placental whole-transcriptome analysis

The underlying pathomechanism in placenta-related selective fetal growth restriction in monochorionic diamniotic twin pregnancy is not known. This study aimed to investigate any differences in placental transcriptomic profile between the selectively growth-restricted twins and the normally grown cotwins in monochorionic diamniotic twin pregnancies. This was a prospective study of monochorionic diamniotic twin pregnancies complicated by selective fetal growth restriction. Placental biopsy specimens were obtained from the subjects in the delivery suite. The placental transcriptome of the selectively growth-restricted twin was compared with that of the normally grown cotwin. This study was divided into 2 stages: (1) gene discovery phase in which placental tissues from 5 monochorionic diamniotic twin pregnancies complicated by selective fetal growth restriction plus 2 control twin pregnancies underwent transcriptome profiling, and transcriptome profiling was carried out using whole-genome RNA sequencing; and (2) validation phase in which placental tissues from 13 monochorionic diamniotic twin pregnancies with selective fetal growth restriction underwent RNA and protein validation. RNA and protein expression levels of candidate genes were determined using quantitative real-time polymerase chain reaction and immunohistochemistry staining. A total of 1429 transcripts were differentially expressed in the placentae of selectively growth-restricted twin pairs, where 610 were up-regulated and 819 were down-regulated. Endoplasmic reticulum lectin and mannose 6-phosphate receptor were consistently differentially up-regulated in all placentae of selectively growth-restricted twins. Quantitative real-time polymerase chain reaction and immunohistochemistry staining were used to validate the results (P<.05). The expression of endoplasmic reticulum lectin and mannose 6-phosphate receptor, which are important for angiogenesis and fetal growth, was significantly increased in the placentae of selectively growth-restricted twin of a monochorionic twin pair.

SUN-635 Profiling of Activation Patterns of Placental mTOR in Pregnancies Complicated by Gestational Diabetes Mellitus

The mammalian target of rapamycin (mTOR) couples’ energy and nutrient abundance to cell growth and is critically involved in the onset and progression of diabetes, cancer and ageing. Placental mTOR is involved in nutrient sensing and transfer to the fetus; animal models suggest that placental mTOR is upregulated in pregnancies complicated by hyperglycaemia (1). In this study we investigated expression patterns and activation of placental mTOR and possible effects of gestational diabetes (GDM). Our study consisted of GDM-mothers (n=28) and their offspring and ii) mothers (n=33) with normal pregnancies (non-GDM) and their infants. Total and phospho-mTOR (Ser2448) expression were determined in placental biopsies using either immunoblotting and immunohistochemistry (IHC) analysis. Newborn anthropometric parameters were also determined at delivery. GDM pregnant women presented with higher fasting glucose levels than non-GDM (98.12±22.82mg/dL; 73.61±9.89mg/dL; p<0.001). No significant difference was found in birth weight or baby length between GDM and non-GDM infants. IHC analysis showed that both total and activated mTOR were predominantly expressed in trophoblasts and to a lesser extend in syncytiotrophoblasts, in both GDM and non-GDM placentas. GDM placentas exhibited a higher mTOR H-score (2) compared to non-GDM (p<0.012), and WB analysis showed a higher phosphor-mTOR signal intensity (p=0.047) in the same group, most likely due to increased total mTOR expression. mTOR expression was also increased in both GDM syncytiotrophoblasts and endothelial cells compared to non-GDM (p<0.001) whereas a reduced signal was detected in stromal phospho-mTOR (p=0.004). No difference was found in trophoblasts or endothelial cells between the 2 study groups suggesting that activation of this kinase is tightly regulated and is relatively independent of changes in total kinase levels. Interestingly bivariate correlation analysis identified an extensive network of significant associations in the expression levels of total, phosphor-mTOR and P/T mTOR between trophoblasts, stroma, endothelial and syncytiotrophoblastsin control placental biopsies; this network was significantly disrupted in GDM placentas, identifying a disheveled regulation of placental mTOR activity. In conclusion, placental mTOR/PmTOR expression is differentially regulated across different placental cell types and is sensitive to hyperglycaemia associated with gestational diabetes mellitus.(1)M. Castillo-Castrejon and TL. Powell. Front Endocrinol (Lausanne). 2017; 8: 306. (2) E. Lakiotaki, et al., Scientific Reports 2016; 6, 21252.

Changes in Vascular Smooth Muscle and Collagen Content in Placental Vessels in Preeclampsia [04L

INTRODUCTION: Preeclampsia results in part from abnormal myometrial invasion by placental spiral arteries. It is unknown if and how fetal vessels remodel to compensate for these changes. We sought to characterize preeclampsia-associated vascular remodeling by comparing vessels within villi of placentas collected from preeclamptic and normotensive patients. METHODS: Carilion Clinic IRB approval was obtained prior to patient enrollment. Singleton pregnancies complicated by preeclampsia and normotensive controls were identified on Labor and Delivery. Placental biopsies were collected following delivery. Smooth muscle (alpha-SMA) and Type III collagen (COLIII) were identified by immunostaining of cryostat sections. Confocal microscopy was used to measure areas of alpha-SMA and COLIII positivity surrounding vessels. Ratios of alpha-SMA or COLIII positivity to vessel area were calculated. These ratios, which represent relative smooth muscle and collagen content, were compared between vessels from preeclamptic and normotensive placentas. RESULTS: Measurements from 342 vessels from 10 placentas (from five preeclamptic patients and five controls) were included in this analysis. Compared to vessels within placentas of normotensive patients, placental vessels of preeclamptic patients demonstrated a significantly reduced area of alpha-SMA positivity (2.48 vs 3.97; P=.008). A trend was seen towards decreased collagen surrounding vessels in preeclamptic placentas, though this was not statistically significant (4.33 vs 5.66; P=.054). CONCLUSION: Preeclampsia is associated with a decrease in vascular smooth muscle within placental villi and may be associated with decreased collagen content. This may represent a mechanism to increase transplacental flow of oxygen in the setting of impaired spiral artery remodeling, but further studies are needed to clarify this relationship.

Effect of sulfadoxine-pyrimethamine doses for prevention of malaria during pregnancy in hypoendemic area in Tanzania

BackgroundMalaria in pregnancy increases the risk of deleterious maternal and birth outcomes. The use of ≥ 3 doses of sulfadoxine-pyrimethamine (SP) for intermittent preventive treatment of malaria (IPTp-SP) is recommended for preventing the consequences of malaria during pregnancy. This study assessed the effect of IPTp-SP for prevention of malaria during pregnancy in low transmission settings.MethodsA cross-sectional study that involved consecutively selected 1161 pregnant women was conducted at Mwananyamala regional referral hospital in Dar es Salaam. Assessment of the uptake of IPTp-SP was done by extracting information from antenatal clinic cards. Maternal venous blood, cord blood, placental blood and placental biopsy were collected for assessment of anaemia and malaria. High performance liquid chromatography with ultraviolet detection (HPLC-UV) was used to detect and quantify sulfadoxine (SDX). Dried blood spots (DBS) of placental blood were collected for determination of sub-microscopic malaria using polymerase chain reaction (PCR).ResultsIn total, 397 (34.2%) pregnant women reported to have used sub-optimal doses (≤ 2) while 764 (65.8%) used optimal doses (≥ 3) of IPTp-SP at the time of delivery. The prevalence of placental malaria as determined by histology was 3.6%. Submicroscopic placental malaria was detected in 1.4% of the study participants. Women with peripheral malaria had six times risk of maternal anaemia than those who were malaria negative (aOR, 5.83; 95% CI 1.10–30.92; p = 0.04). The geometric mean plasma SDX concentration was 10.76 ± 2.51 μg/mL. Sub-optimal IPTp-SP dose was not associated with placental malaria, premature delivery and fetal anaemia. The use of ≤ 2 doses of IPTp-SP increased the risk of maternal anaemia by 1.36-fold compared to ≥ 3 doses (aOR, 1.36; 95% CI 1.04–1.79; p = 0.02).ConclusionThe use of < 2 doses of IPTp-SP increased the risk of maternal anaemia. However, sub-optimal doses (≤ 2 doses) were not associated with increased the risk of malaria parasitaemia, fetal anaemia and preterm delivery among pregnant women in low malaria transmission setting. The use of optimal doses (≥ 3 doses) of IPTp-SP and complementary interventions should continue even in areas with low malaria transmission.

Subsequent pregnancy outcomes according to the presence of acute histologic chorioamnionitis in women with spontaneous preterm delivery.

ObjectiveTo compare subsequent pregnancy outcomes according to the presence of acute histologic chorioamnionitis (HCA) in women with spontaneous preterm delivery (SPTD).MethodsAmong 1,706 women who gave birth twice or more at our institution, 138 women delivered spontaneously at preterm (<37.0 weeks). Subsequent deliveries occurred at our institution and placental biopsy results were available. The study population was categorized into 2 groups based on the presence of acute HCA at the time of SPTD: HCA group (n=52) and non-HCA group (n=86). The primary outcome measures were gestational age at delivery, birthweight, and frequency of preterm delivery in subsequent pregnancies.ResultsThe median gestational age at the time of SPTD was 34.0 weeks (interquartile range [IQR], 28.9–35.3 weeks), and the frequency of acute HCA was 52/138 (38%). There were no differences in gestational age at delivery, birthweight, and frequency of preterm delivery between the HCA group and non-HCA group (median gestational age at delivery, 38.0 weeks (IQR, 36.7–38.8 weeks) in the HCA group vs. 37.9 weeks (IQR, 35.7–39.0 weeks) in the non-HCA group; frequency of preterm delivery, 14/52 (27%) in the HCA group vs. 33/86 (38%) in the non-HCA group; and median birthweight, 3.14 kg (IQR, 2.64–3.45 kg) in the HCA group vs. 2.95 kg (IQR, 2.44–3.36 kg) in the non-HCA group; P>0.1 for all.ConclusionThe presence of acute HCA in women at prior SPTD did not significantly affect their subsequent pregnancy outcomes.

Immune cell infiltrate at the utero-placental interface is altered in placenta accreta spectrum disorders.

To characterize the immune cell infiltrate associated with maternal vascular remodeling in 12 cases of cesarean hysterectomy from women with placenta accreta spectrum (PAS) disorder. Myometrial vessels were evaluated by hematoxylin and eosin and immunohistochemistry stains on tissue microarrays that included samples from the myometrium, deep to the implantation site. Vessels were quantified based on physiologic conversion and immune cell infiltrates. Placental bed biopsies from cases of repeat cesarean section, and decidual vessels from delivered non-PAS placentas were used as controls. Immune cells, predominantly macrophages and T-cells, were present as a band along the placental-myometrial interface in PAS cases. However, within the myometrium, the infiltrate showed a perivascular accentuation. The infiltrates around and within vessel walls were composed of T-cells (CD3) and macrophages (CD68), with fewer NK (CD56), Treg (FoxP3) and rare B-cells (CD20). Plasma cells (CD138) were absent. The majority of vessels with immune cell infiltrates had undergone complete or partial physiologic conversion by trophoblast. However, a subset of unconverted vessels in the myometrium had a similar immune cell infiltration. Control blood vessels showed a similar pattern of leukocytes infiltration in the decidua and placental bed biopsies, but with a lower density. Our findings suggest that myometrial vascular changes in PAS resemble the physiological changes of vessels noted in the implantation site of normal pregnancies. The presence of perivascular immune cell infiltrates in the absence of adjacent trophoblast suggests that the process may be initiated by paracrine effects rather than direct contact or endovascular growth of trophoblast.

Variability of iodine concentrations in the human placenta

Iodine is an essential trace element, necessary for the production of thyroid hormones, which play a key role in optimal foetal growth and (neuro-) development. To date, iodine deficiency remains a health burden in many countries. We investigated the variability of placental iodine concentrations within and between individuals. We used 20 mother-neonate pairs from the ENVIRONAGE birth cohort, took samples at three standardized locations of the placentas, pooled and digested them, and determined the iodine concentrations using an ICP-MS method as an alternative for the Sandell-Kolthoff method. The variability between and within the three sample regions was calculated using the intra-class correlation coefficient (ICC) from the variance components of mixed models. With the Friedman test, the differences between placental biopsies were assessed. The ICC showed a higher between-placenta (68.6%) than within-placenta (31.4%) variability. Subsequently, we used our optimized method to determine iodine concentrations in 498 mother-neonate pairs, which averaged 26.1 μg/kg. For 96 mothers, the urinary iodine concentrations were also determined, which showed no correlation with the placental iodine storage, as was expected. Future studies are necessary to explore the effects of these placental iodine concentrations in relation to health outcomes of mother and child at birth and later in life.

Spiral Arterial Morphology in Relation to Hypertension in Pregnancy and Birth Weight by Placental Bed Biopsy: A Retrospective Analysis

Background: Physiological changes in spiral arteries i.e. remodeling occur upto myometrial segment of the vessels and these changes are important for growth of the fetus. However in hypertensive pregnancies and IUGR, these changes are often not seen and arteries remain non-dilated. The aims of the study were to assess the extent of spiral vessel dilatation in hypertensive pregnancies and IUGR cases. Methods: Placental bed biopsy were taken in cases who underwent caesarean section and gave consent for the study, during the study period. 50 cases were included. Morphology and diameters of spiral arteries were assessed by histopathological examination. Arteries who retained the non-pregnant morphology were considered non-dilated and those who showed features of remodeling were considered normal. 14 cases were excluded as no vessel was seen in biopsy specimen. T-test and Chi-square tests were used for analysis. Out of the included 36 cases, percentage of non-dilated vessels was significantly high in gestational hypertension and preeclampsia(p=0.0230) and also significantly high in IUGR cases, (p=0.0113),irrespective of hypertension. Also the percentage of non-dilated vessels was higher in nulliparous women, though not significant. Placental bed biopsy specimens have limitations because they only provide information about a small segment of the placental bed. It is possible that areas close to the nonbiopsy site may have a completely different degree of vascular transformation. Moreover the number of cases in this study is less. Conclusion: Absence of spiral arterial remodeling is a crucial factor in hypertension in pregnancy and low birth weight babies. And adequate research is needed in this aspect so that such knowledge can help us to prevent hypertension in pregnancy and IUGR.

High Incidence of Herpes Simplex Virus-1 in Cord Blood and Placenta Infection of Women in Southern Brazil.

Estimate the prevalence of human herpesvirus type 1 HSV-1 DNA in placental samples, its incidence in umbilical cord blood of newborns and the associated risk factors. Placental biopsies and umbilical cord blood were analyzed, totaling 480 samples, from asymptomatic parturients and their newborns at a University Hospital. Nested polymerase chain reaction (PCR) and gene sequencing were used to identify the virus; odds ratio (OR) and relative risk (RR) were performed to compare risk factors associated with this condition. The prevalence of HSV-1 DNA in placental samples was 37.5%, and the incidence in cord blood was 27.5%. Hematogenous transplacental route was identified in 61.4% from HSV-1+ samples of umbilical cord blood paired with the placental tissue. No evidence of the virus was observed in the remaining 38.6% of placental tissues, suggesting an ascendant infection from the genital tract, without replication in the placental tissue, resulting in intra-amniotic infection and vertical transmission, seen by the virus in the cord blood. The lack of condom use increased the risk of finding HSV-1 in the placenta and umbilical cord blood. The occurrence of HSV-1 DNA in the placenta and in cord blood found suggests vertical transmission from asymptomatic pregnant women to the fetus.

PREGNANCY, COMPLICATED BY PREECLAMPSIA: FETOPLACENTAL COMPLEX IMMUNE DEADAPTATION AND HISTOSTRUCTURAL FEATURES

The aim: to study and compare the features of the interleukins levels and morphological changes of placenta at various stages of preeclampsia. Materials and methods: 109 pregnant women with preeclampsia of varying severity (study group) and 30 pregnant women with uncomplicated pregnancy (control group) were examined. Immunohistochemical method, proinflammatory interleukins levels, morphological and morphometric analysis of peripheral and central placental areas biopsies on the optical and electron-microscopic level have been used. Results: Morphofunctional changes in the placenta in case of preeclampsia and the increase in the expression level of the transforming growth factor have a series of regular stages from the formation, strain and disruption of adaptive mechanisms with more pronounced signs of morphological immaturity of parenchymal and stromal elements of the placenta, especially in the area of syncytiotrophoblast and spiral vessels. The degree of clinical manifestation of preeclampsia has a correlation relationship with IL-10 deficiency and with the increase in TNF-α, stimulation of macrophage-protein production that contributes to the change in the ratio of Thl / Th2, which are antagonists and inhibit each other's development. Conclusions: The severity of the preeclampsia course correlates with the state of morphofunctional changes in the placenta and changes in the ratio of the pro- and anti-inflammatory interleukins.

Pathomorphological features of the placentas and the placental sites after in vitro fertilization with a donor egg

To identify the structural and immunohistochemical features of the placentas and the placental sites after in vitro fertilization (IVF) with a donor egg (surrogate motherhood). Morphological and immunohistochemical studies were performed on the placental (a placental disk) and placental bed materials obtained after caesarean delivery. The investigation enrolled 26 patients whose pregnancy occurred with IVF with a donor egg according to the surrogacy (IVF-S) program. A comparison group included 13 patients whose pregnancy occurred after IVF with their own eggs. An immunohistochemical study was conducted on paraffin sections made from biopsy material; mouse antibodies to total cytokeratin (clone AE1/AE3, 'Dako'), HLA-DR (clone TAL.1B5, 'Dako'), and CD138 (clone MI15, 'Dako') were used as primary antibodies. The histological examination of the placentas in the IVF-S group showed the high incidence of central ischemic heart attacks (69%), dissociated cotyledon development (61%), pathological villous immaturity mainly with the predominance of intermediate differentiated villi (46%), and massive perivillous fibrinoid deposition (73%). The obtained differences between with the study and comparison groups were significant (p<0.05). The IVF-S group was characterized by the development of lymphoplasmacytic deciduitis (1.23±0.4 and 0.5±0.3 scores). Examination of the placental site biopsy material in the IVF-S group revealed the following changes: remodeling of the spiral arteries was incomplete in more than 40% of cases, and in 30% of the spiral arteries had no gestational changes. In the comparison group, more than 90% of the spiral arteries were characterized by complete remodeling during pregnancy. There was also an increase in the count of multinucleated trophoblastic giant cells (104.56±4.21 and 65.67±14.45) and HLA-DR positive cells (41.86±5.32 and 29.00±1.87). The placentas and the placental sites of the women whose pregnancy occurred with IVF-S are characterized by the development of high lyoplasmacytic deciduitis activity and pronounced placental immune alterations manifested by the high incidence of immune responses at the sites of the closest contact between maternal and fetal tissues. The placental bed exhibited defective spiral artery remodeling, development of chronic inflammatory lesions in the perivascular areas, and an increase in the counts of HLA-DR positive cells and multinucleated trophoblastic giant cells.

27: Maternal inositol supplementation modulates placental metabolic programming in a pregnant murine model of metabolic syndrome

Metabolic syndrome (MS) and hypertension (HTN) in pregnancy impact offspring health in adult life. We showed that offspring born to MS dams have an altered metabolic phenotype that was improved by maternal inositol (INO) supplementation. Our objective was to test the hypothesis that the beneficial effect of INO is mediated through placental metabolic pathways in offspring born to a murine model of MS. Female heterozygous for endothelial nitric oxide synthase gene (eNOS-/+) with hypertension were placed on control (HTN) or high-fat diet for 4 wks to induce a MS phenotype, then bred with wild-type male. On gestational day 1, dams with MS were allocated to receive INO (MS-INO) or water (MS). Three groups were obtained: HTN, MS, and MS-INO. Dams were euthanized at term. Placental biopsy from fetal side were genotyped for eNOS allele and gender. Male (M-PL) and female (F-PL) eNOS-/+ placenta were selected. Glycogen (Gly) was measured by ELISA. Proteins involved in glucose uptake (GLUT4 and IR-β), glycogen synthesis (Akt, pAktT-308 and GSK3), and ATP production (pPDH) were measured using Western blot. 1-way or 2-way ANOVA were used for statistical analysis. Gly was higher in M-PL and F-PL of MS dams vs. HTN dams, and reduced by INO supplementation (Fig. 1A). GLUT4 was higher in M-PL of MS vs. HTN and INO treated dams; no differences were seen in female (Fig. 1B). pPDH was reduced in placenta of M-PL and F-PL of HTN and MS dams, and was increased by INO supplementation (Fig. 1C). IR-β was higher in M-PL vs. F-PL. Akt was not different between groups in either gender. Active pAktT-308 was higher in F-PL from MS vs. HTN dams and was lowered by INO treatment, but no differences were noted in M-PL. GSK3 was higher in M-PL and F-PL of HTN and MS dams and was lowered by INO (Table 1). MS during pregnancy affects offspring placental glucose metabolism by altering its storage and utilization. Male and female adapt differently to those maternal metabolic abnormalities. INO supplementation in MS dams enhances placental glucose use for energy production.View Large Image Figure ViewerDownload Hi-res image Download (PPT)

Chromosomal disorders in the fetus of pregnant woman from Aktau

The article presents the data of cytogenetic study on the fetal karyotype of 310 pregnantwomen from the city Aktau in 2013-2015. Chorionic biopsy and placental biopsy were carried out for248 pregnant women with amniocentesis in 18 cases and cordocentesis in 44 cases. Clinical indicators imposing invasive procedures to pregnant women at risk were maternal age factor, fetal ultrasoundmarkers, serum blood markers determined in pregnant women, the presence of children with multiplecongenital malformations, chromosomal pathologies, etc. Distribution frequencies of fetal chromosomalabnormalities have been determined based on clinical and laboratory studies. The highest frequency ofchromosomal abnormalities of the fetus was indicated in pregnant women with three salient indicators:the factor of the age of the pregnant woman, parameters of biochemical screening, and ultrasound markers. Chromosomal abnormalities of the fetus were detected in 94 (30.3%) pregnant women including 92cases (97.9%) represented by numerical chromosome disorders and 2 cases (2.1%) of structural disorders. Disorders of the autosome system were observed 9.4 times more often comparing to abnormalitiesin the sex chromosome system. Of the numerical chromosome abnormalities, a high specific weight isoccupied by trisomy of the 21-st chromosome which has reached 65.1%. A comparative analysis of thefrequencies of chromosomal abnormalities of the fetus of two port cities has demonstrated a 1.7-foldincrease in the city of Aktau (Kazakhstan) compared with the city of Murmansk (Russia), and the averagefrequencies of fetal karyotype anomalies have made up 19.6% and 11.6%, respectively. Higher level offrequencies for fetal karyotype anomalies in Aktau is possibly associated with unfavorable environmentalconditions in this city caused by the allocation of the oil and gas industry, the repository for tailing toxicand radioactive wastes, and abandoned uranium mines.Key words: karyotype, fetus, prenatal diagnosis, fetal chromosome disorders, ecological state ofurban agglomerations.

False negative non-invasive prenatal screening result for trisomy 18: a case report

We hereby reported a case of false negative non-invasive prenatal screening (NIPS) for trisomy 18. The fetus with increased nuchal translucency (3.2 mm) detected by ultrasound scan at 13+4 gestational weeks received NIPS and the result was negative in chromosomes 21, 18 and 13. A routine ultrasound examination at 22 weeks of gestation revealed multiple anomalies and a second NIPS was offered, which showed a negative result again. The pregnancy was terminated at 22+3 weeks. Multiple fetal and placental biopsies were collected for chromosome analysis using copy number variation sequencing based on high-throughput sequencing and fluorescence in situ hybridization. The fetal karyotype was shown to be 47,XY,+18 in fetal tissues (skin and liver) and umbilical cord, while no chromosomal abnormalities was detected at or near the center of the fetal and maternal surface of the placenta. Results of the chromosomal analysis along the edges of the fetal and maternal surfaces of the placenta were Chr18:47,XY,+18[60]/46,XY[40] and Chr18:47,XY,+18[35]/46,XY[65], respectively. We inferred that placental mosaicism was the cause of the false negative NIPS result. Therefore, genetic counseling before and after NIPS is necessary. Follow-up ultrasound is important for NIPS-negative patients. Invasive prenatal diagnosis is recommended when abnormal ultrasound markers with possible genetic etiology were recognized. Key words: Trisomy 18 syndrome; Ultrasonography, prenatal; False negative reactions; Nuchal translucency measurement

Quantification of spiral artery remodelling using an Adobe Photoshop-based technique.

Uterine spiral arteries undergo remodelling in normal pregnancy, with replacement of the musculoelastic arterial media by fibrinoid containing extravillous trophoblast cells. Deficient spiral artery remodelling is associated with several adverse pregnancy outcomes. Although there are distinct components of spiral artery remodelling, assessment is subjective and often based on an overall impression of morphology. We aimed to develop a quantitative approach for assessment of uterine spiral artery remodelling. Placental bed biopsies were immunostained using smooth muscle markers, digital images of spiral arteries were captured and Adobe Photoshop was used to analyse positive immunostaining. The method was then used to investigate variation in the same vessel at different levels within a paraffin block, and the effect of parity, pre-eclampsia or miscarriage on vascular smooth muscle cell content. Results were also compared with a more subjective morphology-based assessment system. There was good intra- and interobserver agreement and the method correlated well with the more subjective assessment system. There was an overall reduction in vascular smooth muscle, as detected by caldesmon 1 (h-caldesmon) immunopositivity, with increasing gestational age from 8 weeks to term. A previous pregnancy did not affect the amount of spiral artery smooth muscle. Comparison of pre-eclampsia and late miscarriage samples with controls of the appropriate gestational age demonstrated increased medial smooth muscle in pathological samples. This technique provides a simple, rapid, reproducible and inexpensive approach to quantitative assessment of spiral artery remodelling in normal and pathological human pregnancy, a process which although fundamental for successful pregnancy, is still incompletely understood.

DNA methylation profiles of genes associated with angiogenesis in the samples of placenta in pregnancies complicated by intrauterine growth restriction

Background Impairment in placental angiogenesis is blamed for the etiopathogenesis of intrauterine growth restriction (IUGR). Aim To assess the genes related to angiogenesis in placental biopsies of pregnancies complicated by IUGR that could be aberrantly methylated and adversely affect placental angiogenesis. Methods The methylation profiles of soluble fms-like tyrosine kinase-1 (sFLT-1), vascular endothelial growth factor (VEGF), and the placental growth factor (PIGF) were evaluated using Illumina MiSeq™ System in placental biopsies from term IUGR pregnancies without preeclampsia (n = 18) and healthy controls (n = 17). DNA was isolated from samples of tissue collected from the fetal side of the placenta. In the targeted regions, we have identified 30, 24, and 29 CpG islands (CpGi) within sFLT-1, VEGF and PIGF genes, respectively. CpGi which are most methylated in the promoter regions of three genes were selected for the study from the database http://www.ensembl.org. Result(s) IUGR fetuses had significantly lower placental and fetal birth weight than controls. The promoter of sFLT-1 at three CpGi and VEGF at six CpGi were the regions with significant methylation differences between IUGR and control placentas. sFLT-1 was hypermethylated at 265 and 352 CpGi; however, hypermethylation was lower in IUGR group compared to control group at this position. sFLT-1 was hypomethylated at 456 CpGi in IUGR group and hypermethylated at the same region in control group. VEGF was hypomethylated at 668, 703, and 710 CpGi in control and IUGR groups; however, hypomethylation at these positions was significantly higher in control group compared to IUGR. 776, 845, and 863 CpGi of VEGF promoter were significantly hypermethylated in IUGR group whereas hypomethylated in control group. The methylation profile of PIGF did not differ between the groups. After adjustment for the factors known to affect fetal birth weight, DNA methylation of VEGF 668 CpGi had a significant negative association with fetal birth weight in the control and the IUGR group and a positive association with IUGR pregnancies. Conclusion(s) Our results do not support the hypothesis that altered DNA methylation in the placental angiogenic genes is a major mechanism generally involved in IUGR. Only a specific region (at 668 CpGi) corresponding to the promoter of VEGF may serve as an epigenetic marker of IUGR and may be involved in the mechanism of IUGR. Large sample-sized studies are needed to understand the effects of DNA methylation on placental gene function and how they might influence fetal growth.

Air pollution can cross the placenta

Exposure to ambient air pollution is associated with low birth weight, preterm birth, and other adverse birth outcomes. For years, scientists have puzzled over the mechanism behind these negative effects. Now, new research has uncovered an important clue: small particulate matter can cross over the placenta (Nat. Commun. 2019, DOI: 10.1038/s41467-019-11654-3). The study focused on black carbon—minute particles produced by incomplete combustion of fossil fuels and other organic substances. Black carbon particles are especially harmful because they are often coated with toxic substances like heavy metals, says Hasselt University biophysicist Hannelore Bove, who led the study. The researchers took placental biopsies from 28 mothers. When illuminated with short, fast laser pulses, the black carbon particles embedded in the placenta glowed white. Those particles were detected on both the maternal and fetal sides of all placentas sampled. The researchers also found that black carbon concentrations were higher i...

Placental NEGR1 DNA methylation is associated with BMI and neurodevelopment in preschool-age children

ABSTRACTStudies have linked maternal pre-pregnancy obesity and hyperglycaemia with metabolic and neurodevelopmental complications in childhood. DNA methylation (DNAm) might enable foetal adaptations to environmental adversities through important gene loci. NEGR1 is involved in both energy balance and behaviour regulation. The aim of this study was to investigate associations between placental DNAm at the NEGR1 gene locus and childhood anthropometric and neurodevelopmental profiles in preschoolers. We analysed 276 mother-child dyads from Gen3G, a prospective birth cohort from Sherbrooke. At 3yo (40.4 ± 3.0 months), we measured body mass index (BMI) and the mothers reported on offspring neurobehavior using the Strengths and Difficulties Questionnaire (SDQ). We quantified DNAm levels at 30 CpGs at the NEGR1 locus using the MethylationEPIC Array in placental biopsies. DNAm at four CpGs located before NEGR1 second exon predicted child’s BMI z-score (cg26153364: β=−0.16 ± 0.04; p=0.008, cg23166710: β=0.14 ± 0.08; p=0.03) and SDQ total score (cg04932878: β=0.22 ± 1.0; p= 3.0x10−4, cg16525738: β=−0.14 ± 0.18; p=0.01, cg23166710: β=−0.13 ± 0.36; p= 0.04), explaining 4.2% (p=0.003) and 7.3% (p= 1.3 x 10−4) of BMI-z and SDQ variances. cg23166710 was associated with both childhood phenotypes and correlated with NEGR1 placental expression (r=−0.22, p=0.04), suggesting its possible functional role. Together, maternal metabolic characteristics during pregnancy with NEGR1 DNAm levels explained 7.4% (p=4.2 x 10−4) of BMI-z and 14.2% (p=2.8 x 10−7) of SDQ variance at 3yo. This longitudinal study suggests that placental NEGR1 DNAm is associated with adiposity and neurodevelopment in preschool children and highlights its potential role in their comorbidity.