Normal Placenta Research Articles

Patterns of Interferon γ Expression and C4d Deposition in Chronic Intervillositis of Unknown Etiology.

The pathogenesis of chronic intervillositis of unknown etiology (CIUE) may involve IFNγ overexpression. This study assesses the extent of IFNγ expression in CIUE by immunohistochemistry and compares it to spontaneous pregnancy losses. C4d deposition is also assessed to see whether IFNγ and C4d might represent separate diagnostic categories. Placenta from first to early second trimester with high grade CIUE (CHG; 17 cases) and low grade CIUE (CLG; 12 cases) is compared to euploid (SPLN; 18 cases), aneuploid spontaneous pregnancy losses (SPLA, 17 cases), normal placenta (NP, 13 cases). Protein level expression of IFNγ and C4d is assessed on whole tissue sections by immunohistochemistry. 35% of CHG and 42% of CLG show some level of IFNγ expression localized to the luminal surface of syncytiotrophoblast. 12% of SPLA and no SPLN or NP cases are IFNγ positive. C4d deposition is seen in 100% of CIUE, 88% of SPLA, 83% of SPLN, and 46% of NP samples. IFNγ overexpression occurs in approximately 40% of CIUE-related pregnancy losses. IFNγ expression restricted to a subgroup of CIUE implies that IFNγ may define a distinct disease process. The non-discriminatory pattern of C4d deposition suggests it is a non-specific phenomenon possibly related to placental damage.

Effects of Diabetes and Hypertension on Placenta

Placental insufficiency is the failure of the placenta to supply nutrients to the fetus and remove toxic wastes; it is a common cause of intra uterine growth restriction (IUGR), fetal distress peri and postnatal mortality. Specifically gestational diabetes and hypertension during pregnancy alters placental architecture which contributes to insufficiency of the placenta. Placenta is the most accurate record of infant’s prenatal experiences. Clinical condition such as hypertension and diabetes that complicate pregnancy leads to significant structural changes in placenta, these histological changes and morphological alterations in placenta lead to insufficiency, resulting in IUGR and fetal distress. In the present study morphology of placentas collected from diabetic and hypertensive pregnancy will be observed, the histo-pathological findings will be recorded and correlated to birth weight, APGAR scores and congenital anomalies of babies and will be compared to the findings in normal placentas also. This study will be beneficial for obstetrician and pediatrician in gathering knowledge about patho genesis of placental insufficiency; it will add to the knowledge on placental morphology and will high light the abnormalities in placental anatomy.

SWAVE 2.0 Imaging of Placental Elasticity and Viscosity: Potential Biomarkers for Placenta-Mediated Disease Detection.

Pregnancy complications such as pre-eclampsia (PE) and intrauterine growth restriction (IUGR) are associated with structural and functional changes in the placenta. Different elastography techniques with an ability to assess the mechanical properties of tissue can identify and monitor the pathological state of the placenta. Currently available elastography techniques have been used with promising results to detect placenta abnormalities; however, limitations include inadequate measurement depth and safety concerns from high negative pressure pulses. Previously, we described a shear wave absolute vibro-elastography (SWAVE) method by applying external low-frequency mechanical vibrations to generate shear waves and studied 61 post-delivery clinically normal placentas to explore the feasibility of SWAVE for placental assessment and establish a measurement baseline. This next phase of the study, namely, SWAVE 2.0, improves the previous system and elasticity reconstruction by incorporating a multi-frequency acquisition system and using a 3-D local frequency estimation (LFE) method. Compared with its 2-D counterpart, the proposed system using 3-D LFE was found to reduce the bias and variance in elasticity measurements in tissue-mimicking phantoms. In the aim of investigating the potential of improved SWAVE 2.0 measurements to identify placental abnormalities, we studied 46 post-delivery placentas, including 26 diseased (16 IUGR and 10 PE) and 20 normal control placentas. By use of a 3.33-MHz motorized curved-array transducer, multi-frequency (80,100 and 120 Hz) elasticity measures were obtained with 3-D LFE, and both IUGR (15.30 ± 2.96 kPa, p=3.35e-5) and PE (12.33 ± 4.88 kPa, p=0.017) placentas were found to be significantly stiffer compared with the control placentas (8.32 ± 3.67 kPa). A linear discriminant analysis (LDA) classifier was able to classify between healthy and diseased placentas with a sensitivity, specificity and accuracy of 87%, 78% and 83% and an area under the receiver operating curve of 0.90 (95% confidence interval: 0.8-0.99). Further, the pregnancy outcome in terms of neonatal intensive care unit admission was predicted with a sensitivity, specificity and accuracy of 70%, 71%, 71%, respectively, and area under the receiver operating curve of 0.78 (confidence interval: 0.62-0.93). A viscoelastic characterization of placentas using a fractional rheological model revealed that the viscosity measures in terms of viscosity parameter n were significantly higher in IUGR (2.3 ± 0.21) and PE (2.11 ± 0.52) placentas than in normal placentas (1.45 ± 0.65). This work illustrates the potential relevance of elasticity and viscosity imaging using SWAVE 2.0 as a non-invasive technology for detection of placental abnormalities and the prediction of pregnancy outcomes.

Application of Digital Imaging and Artificial Intelligence to Pathology of the Placenta.

Digital imaging, including the use of artificial intelligence, has been increasingly applied to investigate the placenta and its related pathology. However, there has been no comprehensive review of this body of work to date. The aim of this study was to therefore review the literature regarding digital pathology of the placenta. A systematic literature search was conducted in several electronic databases. Studies involving the application of digital imaging and artificial intelligence techniques to human placental samples were retrieved and analyzed. Relevant articles were categorized by digital image technique and their relevance to studying normal and diseased placenta. Of 2008 retrieved articles, 279 were included. Digital imaging research related to the placenta was often coupled with immunohistochemistry, confocal microscopy, 3D reconstruction, and/or deep learning algorithms. By significantly increasing pathologists' ability to recognize potentially prognostic relevant features and by lessening inter-observer variability, published data overall indicate that the application of digital pathology to placental and perinatal diseases, along with clinical and radiology correlation, has great potential to improve fetal and maternal health care including the selection of targeted therapy in high-risk pregnancy.

The new landscape of differentially expression proteins in placenta tissues of gestational diabetes based on iTRAQ proteomics

IntroductionGestational diabetes mellitus (GDM) refers to abnormal glucose tolerance that occurs or is firstly diagnosed during pregnancy. GDM is related to various adverse pregnancy outcomes, but GDM pathogeny has not been fully elucidated. Nevertheless, previous studies have observed that many proteins in the placentas of patients with GDM are dysregulated. The present study aimed to establish a novel differentially expressed protein (DEP) landscape of GDM and normal maternal placentas and to explore the possible connection between DEPs and GDM pathogenesis. This study provides new insights into the mechanism of GDM and should make an important contribution to the development of biomarkers. MethodsThe morphological characteristics of the placenta were observed on 30 GDM and normal maternal placental tissues stained with haematoxylin and eosin. Isobaric tags for relative and absolute quantitation (iTRAQ) was used in the proteomics screening of the DEPs of the normal and GDM maternal placentas. Bioinformatics analysis was performed on the DEPs, and parallel reaction monitoring (PRM) was performed to verify the DEPs. Finally, the quantitative analysis of iTRAQ and PRM was verified by immunohistochemical assay. ResultsA total of 68 DEPs in the GDM placenta were identified with iTRAQ proteomics experiment, comprising 21 up-regulated and 47 down-regulated DEPs. Bioinformatics analysis showed that the regulation of transport, catabolic process of non-coding RNA, cytoskeleton and cell binding were the most abundant Gene Ontology terms, and RNA degradation was an important pathway for significant enrichment. Protein–protein interaction network analysis showed that heterogeneous nuclear ribonucleoproteins A2/B1 (HNRNPA2B1), heterogeneous nuclear ribonucleoprotein A/B (HNRNPAB), heterogeneous nuclear ribonucleoprotein L (HNRNPL) and heterogeneous nuclear ribonucleoprotein A3 (HNRNPA3) were the cores of the up-regulated proteins. Band 3 anion transport protein (SLC4A1), spectrin beta chain erythrocytic (SPTB), ankyrin-1 (ANK1), spectrin beta chain non-erythrocytic 2 (SPTBN2), D-3-phosphoglycerate dehydrogenase (PHGDH) and exosome complex component RRP42 (EXOSC7) were the cores of the down-regulated proteins. These proteins are involved in the binding, splicing, processing, transport and degradation of RNA and in the formation and maintenance of the cytoskeleton. PRM verification results showed that seven proteins, namely, epiplakin (EPPK1), cold-inducible RNA-binding protein (CIRBP), HNRNPA2B1, HNRNPAB, HNRNPL, Ras-related protein Rab-21 (RAB21) and Ras-related protein Rab-3B (RAB3B), were up-regulated, whereas SPTB and SLC4A1 were down-regulated. The results of immunohistochemical assay also showed that the expression of five proteins, namely EPPK1, HNRNPA2B1, HNRNPAB, CIRBP and RAB21, were significantly higher in GDM placental tissues (P < 0.01). The GDM placentas showed changes in the morphological evaluation, including poor villous maturation, obvious increase in the number of syncytiotrophoblast nodules, thickening of the wall of dry villous arterioles with lumen stenosis, increased fibrinous exudation and excessive filling of villous interstitial vessels. DiscussionDifferentially expressed proteins related to a variety of biological processes in the GDM placenta were found. Fourteen proteins, namely, HNRNPA2B1, HNRNPAB, HNRNPL, HNRNPA3, EPPK1, CIRBP, RAB21, RAB3B, SLC4A1, SPTB, ANK1, SPTBN2, PHGDH and EXOSC7, which were differentially expressed in the placenta, may play an important role in regulating the occurrence and development of gestational diabetes through multi-channel and multi-link regulation.

Expression of NF-κB and VEGF in normal placenta and placenta previa patients.

Placenta previa is a pregnancy condition associated with the development of complications related to placental insufficiency, including hypertension, preeclampsia and perinatal mortality. Dysfunction in uteroplacental arteries causes the release of cytokines, leukotrienes and immunomodulatory hormones, which leads to an inflammatory reaction. The nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway and vascular endothelial growth factor (VEGF) are known to play crucial roles in inflammation and angiogenetic regulation. This study aimed to demonstrate the morphometric and immunohistochemical effects on inflammation and angiogenesis underlying placenta previa. Twenty pregnant patients with placenta previa and 20 healthy pregnant patients, all between 30 and 38 weeks gestational age, were included in the study. The gestational age of the pregnancies was determined according to the last date of menstruation and/or ultrasonographic measurements. Blood samples and clinical data were obtained from the prenatal patient groups. Samples were taken from the connecting stem region from both groups. The mean difference between the control and placenta previa patients was statistically significant for the parameters of blood vessels in villi, diameter of floating small villus, decidual cells, syncytial knots, congestion in blood vessels, fibrinoid accumulation, and inflammation. Significant degeneration and apoptotic changes in the syncytial cells of the root villi and an increase in syncytial nodes and bridges were observed in the placenta previa specimens. In the connecting stem region of the placenta previa samples, blood vessel dilatation, endothelial cell hyperplasia and a higher number of syncytial nodes were observed. In the immunohistochemical examination of the placenta previa samples, an increase in NF-κB and VEGF expression was observed in the endothelial cells, syncytial cells and Hofbauer cells. Vascular endothelial growth factor was found to stimulate endothelial cell proliferation and migration, and to significantly affect angiogenesis during the developmental process of the placenta and remodeling of the uterine vessels, inducing NF-κB signaling and apoptotic development during cytotrophoblastic invasion in the vascularization of the placenta.

Dominance of Pro-Inflammatory Cytokines Over Anti-Inflammatory Ones in Placental Bed of Creta Cases.

The objective of the study is to investigate changes occurring in key inflammatory cytokines at molecular level (including genetic and protein) in placental bed of placenta creta compared to that of normal placenta and their correlation to interstitial extravillous trophoblasts (EVT) number. Case-control study including placentas of patients with invasive placentation (creta placentas, n = 19) compared with those of normal placentation (n = 19). Besides routine histology and immunocytochemistry detection (cytokeratin-7 [CK-7]), addition to biochemical evaluation of expression of various cytokines including tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), IL6, IL-1RA, IL-8, IL-10, and IL-13 was carried out. Routine histological examination of placentas of creta cases revealed CK-7+ extravillous trophoblasts (EVT) penetrating deeply the myometrium with various histopathological arrangements and trophoblastic vascular invasion of the deep myometrial blood vessels. A significant increase (P < 0.05) in the mRNA expression of TNF-α, IL-1 β, and IL6 with an insignificant decrease in placental bed IL-1RA, IL-8, IL-10, and IL-13 was observed in creta cases compared to the control ones. A corresponding significant increase was detected in the protein levels of TNF-α, IL-1 β, and IL-6 as well as an insignificant decrease in placental bed IL-1RA, IL-8, IL-10, and IL-13 in creta cases compared to the normal ones. Moreover, we displayed a significant positive correlation (P < 0.05) between interstitial EVT number and mRNA expression of almost all pro-inflammatory cytokines with negative but insignificant correlation with anti-inflammatory cytokines in creta cases. The upregulated pro-inflammatory cytokines and the correlation of their expression with the increased interstitial EVT provide a supporting evidence of their potentially more relevant role in the development of placenta creta than the anti-inflammatory ones.

The neutrophil-to-lymphocyte ratio: A low-cost antenatal indicator of placental chorioamnionitis in women who deliver preterm without clinical signs and symptoms of infection

Chorioamnionitis is present in up to 70% of spontaneous preterm births and is associated with poor maternal, fetal and neonatal outcomes. ObjectiveTo explore the relationship between the neutrophil-to-lymphocyte ratio and histological chorioamnionitis in women who delivered preterm with no clinical signs or symptoms of infection. Study designThis was a retrospective analysis of a cohort of women who delivered spontaneously between 16 and 36+6 weeks at a tertiary UK hospital. Only women with placental histology and no signs of clinical infection were included. The neutrophil-to-lymphocyte ratio was calculated from a full blood count sample taken routinely within 24 h of delivery. The neutrophil-to-lymphocyte ratio was also calculated from first trimester booking bloods (<13 + 6 weeks) in a subgroup. Placental histopathology was categorised as either inflammatory (i.e. histologic chorioamnionitis, with or without evidence of fetal inflammatory response) or non-inflammatory (vascular pathology or a normal placenta). Results169 women had available placental pathology and were included in the analysis. 70 % (118/169) had confirmed placental inflammation. The mean neutrophil-to-lymphocyte ratio was significantly raised in this group compared to those with normal (n = 24) or vascular (n = 27) pathology (inflammatory neutrophil-to-lymphocyte ratio 9.81 vs non-inflammatory neutrophil-to-lymphocyte ratio 6.53, p = 0.002.The delivery neutrophil-to-lymphocyte ratio had an area under the receiver operating characteristic curve of 0.69 (0.60 to 0.78) for predicting placental inflammation. A raised neutrophil-to-lymphocyte ratio (>6) was associated with an odds ratio of 5.2 (95 % CI 2.55 to 10.56) for histological chorioamnionitis, with a sensitivity of 80 % and negative predictive value of 86 %. A higher cut-off of 9 had a negative predictive value of 79 % for fetal inflammatory response. ConclusionsA raised neutrophil-to-lymphocyte ratio is associated with a 5-fold increased risk of histological chorioamnionitis in women who delivered early without signs or symptoms of infection. It was also raised at the time of preterm labour compared to the first trimester. A full blood count is an almost universal investigation in women admitted in preterm labour, often repeated, making this inexpensive and non-invasive ratio a useful additional antenatal biomarker in women admitted in spontaneous preterm labour at risk of subclinical chorioamnionitis and its associated poor outcomes.

Downregulation of PDCD4 by deSUMOylation associates with the progression of gestational trophoblastic disease

IntroductionGestational trophoblastic disease (GTD) encompasses a range of trophoblastic disorders from hydatidiform mole (HM), to malignant gestational trophoblastic neoplasia (GTN). The exact molecular mechanisms of GTN remain unknown. Dysregulation and dysfunction of programmed cell death 4 (PDCD4)have been observed in many cancers. The roles of PDCD4 in GTD have not been previously reported. MethodsA total of 161 cases of formalin-fixed, paraffin-embedded trophoblast blocks, and 36 cases of fresh trophoblast tissues were collected, including normal first trimester placentas, HM, and invasive HM. Choriocarcinoma cells JAR and JEG-3 were employed. The expressions of PDCD4 and small ubiquitin-like modifier 2/3 (SUMO2/3) were examined by immunohistochemistry, quantitative reverse transcription PCR and Western blotting in trophoblastic tissues and cells. The relationship between SUMOylation and PDCD4 was investigated. The effects of PDCD4 on proliferation, invasion, and migration of choriocarcinoma cells were evaluated by Cell Counting Kit-8 and transwell assays post siRNA transfection. Extracellular Matrix & Adhesion Profiler PCR Array was used to screen the downstream molecules of PDCD4. ResultsPDCD4 was significantly repressed in HM tissues. Loss of PDCD4 expression was demonstrated in 90% invasive HMs. Choriocarcinoma cells also displayed with suppressed PDCD4 expression. The varied expression of PDCD4 was paralleled by SUMO2/3. Inhibition of SUMOylation reduced the expression of PDCD4. Silencing of PDCD4promoted proliferation/migration/invasion, upregulatedMMP3/MMP8/ITGB2, and downregulated TIMP1/TIMP2 in choriocarcinoma cells. DiscussionOur results suggest that reduced SUMOylation is one reason for suppressed PDCD4 in GTD. Loss of PDCD4 likely determines the malignant phenotype of GTN by dysregulating some members of the MMPs/TIMPs/integrins complex.

ODP434 Maternally-Derived Pancreatic Extracellular Vesicle Encompassed miRNAs Influence Placental Development in Pregnancies Complicated by Gestational Diabetes

Abstract Pregnancies affected by gestational diabetes (GDM) commonly result in large-for-gestational-age (LGA) infants; these infants have an increased risk of developing cardiometabolic complications compared to appropriate-for-gestational-age (AGA) infants. The mechanisms responsible are unclear but GDM/LGA is associated with altered placental development/function. We have recently reported altered levels of extracellular vesicle (EV) encapsulated miRNAs, including miR-375 (a pancreatic-specific miRNA) in maternal serum prior to the onset of LGA. We propose that the EVs containing miR-375 are produced by maternal pancreas, and that increased miR-375 in maternal circulation contributes to LGA by influencing placental development/function. Human placental tissue was collected from GDM pregnancies that delivered AGA (n=14) or LGA infants (n=10). Levels of mature miR-375 and primary transcript (pri-miR-375) were measured by QPCR. Mature miR-375 was increased in GDM-LGA (4.74-fold; p&amp;lt;0. 01) compared to GDM AGA placentae. Mature miR-375, but not pri-miR-375, was detected in human placenta suggesting that miR-375 is not produced in the placenta but instead is transported, potentially via EVs in maternal circulation. Intact human pancreatic islets (20,000 IEQ/patient) were obtained through the Integrated Islet Distribution Program (IIDP) from female donors of reproductive age. Islets were cultured in EV-depleted media containing 5.5mM (normoglycemia) or 7mM (mild hyperglycaemia) glucose for 3 days. Islet purity and viability were confirmed using dithizone incorporation and trypan blue exclusion, respectively. Media was collected at 24-hour intervals, pooled from all time points, concentrated and EVs were isolated by size exclusion chromatography. Confirmation of the presence of EVs was confirmed by electron microscopy (typical cup-shape morphology of EVs), nanoparticle tracking analysis (NTA; mean size of 83.5-163.2nm; concentration of 1.28×109-1.82×1010 particles/ml), Western blotting (EV-enriched proteins). QPCR confirmed that mature miR-375 was present in islet-derived EVs and revealed that levels were altered following culture in 7mM glucose compared to 5.5mM. Islet-derived EVs were labelled with maleimide-488 and uptake into explants of normal human term placenta was confirmed using fluorescent microscopy=3). miR-375 was overexpressed in normal human placental explants (by 30-fold, p&amp;lt;0. 05; n=6) using specific mimics (100nM for 72 hours) and TMT mass spectrometry was performed to assess the impact of miR-375 overexpression on the placental proteome. Functional enrichment analysis (FEA) was performed on differentially expressed proteins (DEP; P&amp;lt;0. 05; -0.41≥log2foldchange≥0.58) using over representation analysis and Ingenuity Pathway Analysis. This revealed that DEGs were significantly enriched in pathways associated with placental growth, mitochondrial function and glucose metabolism. Our data demonstrates that miR-375 is increased in the placenta in GDM/LGA and that this influences placental development/function. miR-375 does not appear to be produced in the placenta but instead, is likely released in EVs from maternal pancreas under conditions of hyperglycaemia and transported to the placenta, via maternal circulation. This provides insight into the potential mechanisms contributing to LGA in pregnancies complicated by GDM. Presentation: No date and time listed

Regulatory Effect of Vitamin D on Renin Expression at Maternal-Fetal Interface

To investigate the regulatory effect and mechanism of vitamin D on the local renin-angiotensin system at maternal-fetal interface in the pathological process of preeclampsia (PE). The mRNA and protein expression of renin in decidua of normal pregnancy and PE placentas was determined by RT-PCR and Western blot. Normal decidual tissues were treated with active and inactive vitamin D for 48 h in vitro and the expressions of renin and vitamin D deactivating enzyme CYP24A1 were determined by RT-PCR and Western blot. Normal decidual stromal cells and glandular epithelial cells were isolated and purified, and identified by immunocytochemical staining. RT-PCR was used to examine the mRNA of vdr, cyp27 b1, cyp24 a1, and renin in the two types of cells and in decidual tissue, and the mRNA products were subjected to gel electrophoresis. These two cell types were treated with active and inactive vitamin D in vitro and the expressions of renin and vitamin D deactivating enzyme CYP24A1 were determined by RT-PCR and Western blot. Decidual gland epithelial cells were treated with protein kinase A (PKA) activator forskolin or inhibitor H89 to explore the interaction between PKA pathway and vitamin D in the regulation of renin expression. The expression of renin in PE decidua was significantly higher than that of normal control at transcriptional and translational levels ( P<0.05). Vitamin D treatment could significantly down-regulate the expression of renin in normal decidua tissues ( P<0.05), while it significantly up-regulated CYP24A1 expression ( P<0.001). Decidual stromal cells and gland epithelial cells were successfully isolated from decidual tissue. Compared with that in decidual stromal cells, the mRNA level of vitamin D-related molecules in gland epithelial cells was more similar to that in decidual tissue. Active or inactive vitamin D treatment significantly inhibited the expression of renin in glandular epithelial cells ( P<0.05), but the expression of renin in decidual stromal cells was not affected. However, the treatment of active or inactive vitamin D in these two kinds of cells significantly increased the expression of CYP24A1 ( P<0.001). Active vitamin D could significantly inhibit the upregulation of renin by PKA agonist forskolin, and could inhibit the expression of renin through synergy with PKA inhibitor H89. The expression of renin in placental decidua is up-regulated in patients with PE, and the activation of local renin-angiotensin system at the maternal-fetal interface may be involved in the pathogenesis of PE. Vitamin D can specifically down-regulate renin expression in human decidual gland epithelial cells by competing with the PKA pathway. Vitamin D supplementation may have potential value for clinical intervention of PE.

The Blocking of Integrin-Mediated Interactions with Maternal Endothelial Cells Reversed the Endothelial Cell Dysfunction Induced by EVs, Derived from Preeclamptic Placentae.

Placental extracellular vesicles (EVs) have increasingly been recognized as a major mediator of feto-maternal communication. However, the cellular and molecular mechanisms of the uptake of placental EVs by recipient cells are still not well-understood. We previously reported that placental EVs target a limited number of organs in vivo. In the current study, we investigated the mechanisms underlying the uptake of placental EVs into target cells. Placental EVs were derived from explant cultures of normal or preeclamptic placentae. The mechanisms underlying the uptake of placental EVs were elucidated, using the phagocytosis or endocytosis inhibitor, trypsin-treatment or integrin-blocking peptides. The endothelial cell activation was studied using the monocyte adhesion assay after the preeclamptic EVs exposure, with and/or without treatment with the integrin blocking peptide, YIGSR. The cellular mechanism of the uptake of the placental EVs was time, concentration and energy-dependent and both the phagocytosis and endocytosis were involved in this process. Additionally, proteins on the surface of the placental EVs, including integrins, were involved in the EV uptake process. Furthermore, inhibiting the uptake of preeclamptic EVs with YIGSR, reduced the endothelial cell activation. The interaction between the placental EVs and the recipient cells is mediated by integrins, and the cellular uptake is mediated by a combination of both phagocytosis and endocytosis.

Placental proteome in late‑onset of fetal growth restriction.

Fetal growth restriction (FGR) occurs when the fetus does not reach its genetically programmed intrauterine potential for growth and affects ~5–10% of pregnancies. This condition is one of the leading causes of perinatal mortality and morbidity associated with obstetric and neonatal complications. Placental dysfunction in FGR causes an impairment in the transfer of nutrients and oxygen from the mother to the developing fetus. Maternal adaptations to placental insufficiency may also play a role in the pathophysiology of FGR. The present study aimed to compare the proteome of the placentas of 18 women with the physiological course of pregnancy and eutrophic fetus [estimated fetal weight (EFW) >10th percentile; control group] and 18 women with late FGR (EFW <10th percentile) diagnosed after 32 weeks of pregnancy, according to the Delphi consensus (study group). The U. Mann-Whitney test was used to compare two independent groups. The R. Spearman correlation coefficient significance test was used to assess the existence of a relationship between the analyzed measurable parameters. P<0.05 was considered to indicate a statistically significant difference. The tests showed the presence of 356 different proteins which were responsible for the regulation of gene transcription control, inhibiting the activity of proteolytic enzymes, regulation of trophoblast proliferation and angiogenesis and inflammatory response. In the FGR placental proteome, other detected proteins were mostly involved in response to oxidative stress, cellular oxidation and detoxication, apoptosis, hemostatic and catabolic processes, energy transduction protein interactions, cell proliferation, differentiation and intracellular signaling. The present study used chromatographic mass-spectrometry to compare the placental proteome profiles in pregnancies complicated by late-onset FGR and normal pregnancy. Comparative analysis of proteomes from normal and FGR placentas showed significant differences. Further research is needed to clarify maternal and fetal adaptations to FGR.

Role of Ultrasound in Early Detection of Molar Pregnancy

OBJECTIVES The objective of this study was to investigate the role of ultrasound in the detection of early pregnancies presenting with placental molar changes. METHODOLOGY This retrospective study was carried out at Radiology department of Hayatabad Medical Complex, Peshawar, between Aug 2020 and July 2021. All cases suspected of molar changes were evaluated from 10-14 weeks of gestation. The patient were referred to our department because of fetal abnormality, maternal disease or age and family or previous pregnancy history or were identified from an ongoing ultrasound screening study for aneuploidy by measurement of fetal nuchal translucency thickness. Transabdominal or transvaginal sonography was used to examine the fetus and placenta. RESULTS Total 85 patients of molar pregnancies were enrolled in the study. Age ranged between 20-45 years with a mean age of 32.5 years. There were 60(70.6%) complete moles (CM) and 25(27.4%) partial moles (PM) were suspected on ultrasound. In case of complete mole coexisting with a normal singleton or twin pregnancy, the molar placenta was clearly separated from the normal placenta, whereas with partial moles the molar structures were dispersed inside the placental mass. CONCLUSION Ultrasound detection of molar pregnancy remains a diagnostic challenge. Data suggest that there has been an increase in both the predictive value and the sensitivity of ultrasound over time; however, the diagnostic criteria remain ill defined. Prior to managing a miscarriage, being aware of the possibility of molar pregnancy will guide treatment and allow for adequate follow-up.

Texture analysis of sonographic image of placenta in pregnancies with normal and adverse outcomes, a pilot study

IntroductionMany maternal and fetal morbidity and mortality from complications of pregnancy have been attributed to placenta abnormality. Placenta assessment in developing countries is mainly through ultrasonography which is subjective and prone to error. Objective assessment of placental abnormalities through texture analysis has been frequently done using magnetic resonance images with little done on ultrasound generated images, thus, the need for this study. The study is aimed at using statistical texture analysis in characterizing placenta tissue into normal and abnormal placenta as well as testing the accuracy of different texture analysis algorithms in differentiating placenta into normal and abnormal placental tissues. MethodsThis longitudinal study involved 500 ultrasound-generated placenta images from patients screened for adverse pregnancy outcomes in a private hospital in Enugu. These images were loaded onto an HP laptop for viewing. Two regions of interest were selected from the placenta tissue where texture features were extracted and were classified into normal and abnormal placentas using MaZda® software version 47 while the accuracy of the classification descriptors was assessed using WEKA classification algorithms. ResultsCo-occurrence matrix, run length matrix and histogram parameters differentiated normal placenta tissue from abnormal placental tissues (p-value <0.05) while variance is the only absolute gradient parameter that can differentiate normal placenta tissue from abnormal placenta tissues. All feature descriptors show high classification accuracy using KNN and ANN algorithms. ConclusionTexture analysis can differentiate normal placenta tissues from abnormal placenta tissue which will reduce the errors associated with subjective assessment of the placenta echogenicity. Implications for practiceIntegrating these computer-aided algorithms into our ultrasound machines will lead to early detection of abnormal placenta tissues as early management results in better pregnancy outcomes.

Twin pregnancy with complete hydatidiform mole and co-existing fetus: A report of 15 cases with a clinicopathological analysis and DNA genotyping.

Complete hydatidiform mole (CHM) with co-existing fetus (CHMCF) is very uncommon. In this study, we investigated the clinicopathological features and DNA genotype in 15 CHMCF. Seven patients (46.7%) developed post-molar gestational trophoblastic disease (GTD), 5 of which had lung metastasis. CHMCF was histologically characterized by a mixed pattern of CHM and the non-molar placenta, mimicking partial hydatidiform mole and placental mesenchymal dysplasia. p57 immunostaining showed a divergent staining pattern, positive in the normal placenta and negative in the CHM component. DNA genotyping of the CHM villi demonstrated exclusively paternal alleles consisting of homozygous/monospermic (n=9) and heterozygous/dispermic patterns (n=5) at multiple informative loci. We conclude that CHMCF confers a high risk for post-molar GTD. DNA genotyping contributes significantly to the precision diagnosis of CHMCF.

PD-L1 enhances migration and invasion of trophoblasts by upregulating ARHGDIB via transcription factor PU.1

As the main constituent cells of the human placenta, trophoblasts proliferate, differentiate, and invade the uterine endometrium via a series of processes, which are regulated exquisitely through intercellular signaling mediated by hormones, cytokines, and growth factors. Programmed cell death ligand 1 (PD-L1) is a biomarker of the response to immune checkpoint inhibitors and can regulate maternal-fetal immune tolerance during pregnancy progression. Recently, it was found that PD-L1 may regulate obstetric complications by affecting the function of trophoblasts. Therefore, we examined the expression and localization of PD-L1 in the human placenta and observed the effects of PD-L1 on trophoblasts migration and invasion in both the trophoblasts line HTR-8/SVneo and an extravillous explant culture model. Finally, we explored the molecular mechanisms underlying PD-L1-regulated trophoblasts migration and invasion through RNA sequencing and bioinformatics analysis. Our data showed that PD-L1 was mainly expressed in syncytiotrophoblasts and that its protein levels increased with gestational age. Interestingly, the protein expression of PD-L1 was significantly decreased in placentas from pregnancies with preeclampsia compared with normal placentas. Importantly, the migration and invasion abilities of trophoblasts were significantly changed after knockdown or overexpression of PD-L1 in HTR-8/SVneo cells and an extravillous explant culture model, which was partially mediated through the transcription factor PU.1 (encoded by Spi1)-regulated Rho GDP-dissociation inhibitor beta (ARHGDIB) expression. These results suggested that PD-L1 was highly involved in the regulation of trophoblasts migration and invasion, providing a potential target for the diagnosis and treatment of placenta-derived pregnancy disorders.

The m6A methyltransferase METTL3 promotes trophoblast cell invasion by regulating MYLK expression

IntroductionThe progression of placental diseases such as preeclampsia is closely related to trophoblast dysfunction. Recent studies indicated the dysregulation of N6-methyladenosine (m6A) RNA modification in trophoblast disorders, while the function of METTL3, a methyltransferase of m6A, in trophoblasts remains to be studied. MethodsThe expression of METTL3 was determined by real-time PCR and immunoblotting. METTL3 expression in trophoblast cell lines HTR-8/SVneo and JEG-3 was knocked down using shRNA. The invasion of trophoblast cells in Matrigel was determined using xCELLigence. The m6A-containing transcripts was determined by m6A-sequencing in HTR-8/SVneo cells. The myosin light chain kinase (MYLK) gene was transfected into HTR-8/SVneo cells. ResultsThe expression of METTL3 was downregulated in preeclamptic placentae compared to normal placentae. Knockdown of METTL3 repressed the invasion of extravillous trophoblast cells. Mechanistically, METTL3 promoted the stability of MYLK mRNA through m6A modification. Overexpression of MYLK rescued retarded cell invasion by METTL3 depletion. DiscussionCollectively, our results highlight an essential role of METTL3-MYLK axis in trophoblast invasion.

Correlation of placental thickness and placenta percreta in patients with placenta previa: findings from MRI.

This study aimed to identify if placental thickness measured from MRI images correlated with placenta percreta in patients with placenta previa. Placental thickness was retrospectively measured in 161 patients from July 2018 to August 2020. The measurements were performed at the thickest part of the placenta in the lower uterine segment on the mid-sagittal plane MR images by two independent radiologists. Intraoperative and pathologic findings were the standard of reference. Univariate and multivariate analyses were performed to identify the relationship between clinical features, placental thickness, and placenta percreta. The predictive ability of placental thickness was demonstrated using receiver operating characteristic curve analysis. Placental thickness in patients with placenta percreta was significantly higher than in patients with placenta increta, placenta accreta, and normal placentas (p < 0.05). Multivariate analysis revealed that placental thickness was the only independent risk factor for placenta percreta. The cutoff value of placental thickness was 4.35cm for differentiating placenta percreta in patients with placenta previa. Patients with placenta percreta had the highest placental thickness. Placental thickness was correlated with placenta percreta.

Placental morphology in Covid-19: A study in tertiary hospital in Telangana

Due to emerging new variants of Corona virus, study of placental histopathological changes helps in understanding its tissue tropism and effects of the virus of maternal and foetal morbidities. Prospective Observational Study.To evaluate and describe histopathologic findings in the placentas of women affected with COVID19. A total of 16 placentas from pregnant women who were infected with COVID-19 in any gestational age and delivering between March 2020 and February 2022 were examined and compared with normal third trimester placentas received in the same time period. Among the 16 placentas examined, most significant change as a sign of maternal malperfusion is increased intervillous fibrin deposition, which is seen in 81.25% cases, followed by infarction, calcification, increased syncytial knots, haemorrhage, calcification, vascular congestion and fibrin thrombi. Most significant foetal malperfusion sign is Chorangiosis which is seen in 37.5% cases, followed by thrombi in umbilical vessels. Also, 25% of the placentas examined showed Villitis, 18.75% showed Deciduitis, 6.25% showed Chorioamnionitis and 6.25% showed vasculitis. COVID-19 placentas show increased prevalence of decidual arteriopathy and features of foeto-maternal vascular malperfusion which can be related to the consequences in foetus like Preterm delivery, IUGR and respiratory distress.