Placental Protein Levels Research Articles

Placental Protein 13 and Syncytiotrophoblast Basement Membrane Ultrastructures in Preeclampsia.

Background and Objectives: Preeclampsia has been linked to an inflammatory response that may be brought on by endothelial cell dysfunction. This paper investigates the pathomechanism of syncytiotrophoblast basement membrane (STBM) damage and Placental Protein 13 (PP13) release, which may have a role in systemic endothelial dysfunction in preeclampsia. Materials and Methods: This comparative cross-sectional study involves 54 preeclampsia patients (27 early-onset preeclampsia and 27 late-onset preeclampsia) and 27 pregnant women with normal blood pressure. An enzyme-linked immunosorbent assay was performed to evaluate maternal blood levels of PP13. Following birth, a portion of the placenta was collected for transmission electron microscope (TEM) and immunohistochemical (IHC) analysis. The data were analyzed using STATA version 15. Results: PP13 expression in the placental syncytiotrophoblast was significantly lower in the early-onset preeclampsia, compared to late-onset preeclampsia and normotensive pregnancy, group (p < 0.001). In contrast, serum PP13 levels were found to be the highest in the early-onset preeclampsia group, although no significant difference were found in mean maternal serum levels of PP13 between the three groups. The decreased PP13 expression in placental syncytiotrophoblast can be attributed to the greater extent of damage in the STBM in early-onset preeclampsia that leads to the release of a larger amount of PP13 into maternal circulation. The hypothesis aligns with the TEM analysis results. Preeclamptic pregnancies showed placental syncytiotrophoblast aponeurosis, whereas normotensive pregnancies did not. Placental lesions and STBM shedding were found to be more pronounced in early-onset preeclampsia compared to late-onset preeclampsia. Conclusions: PP13 and STBM damage may play a role in systemic endothelial dysfunction in preeclampsia.

Predictive Capabilities of Human Leukocyte Antigen-G and Galectin-13 Levels in the Amniotic Fluid and Maternal Blood for the Pregnancy Outcome.

Background and Objectives: Increasing evidence points to the significant role of the angiogenic factor levels in screening for pregnancy outcome. To examine the potential relationship between concentrations of placental protein 13 (PP13) and soluble human leukocyte antigen-G (sHLA-G) in maternal serum and amniotic fluid at 16-23 weeks of gestation and the sonographic features of pregnancy as well as pregnancy outcome. Materials and Methods: PP13 and sHLA-G in serum and amniotic fluid, fetal biometrical data, and placental volume and perfusion indices were determined in 71 euploid, singleton pregnancies. Results: The serum sHLA-G level exhibits a negative correlation with the serum PP13 level (r = -0.186, p < 0.001) and a positive correlation with the sHLA-G level in amniotic fluid (r = 0.662, p < 0.001). A significant correlation was found between serum sHLA-G level and placental volume (r = 0.142, p < 0.05) and between amniotic sHLA-G level and placental perfusion (r = -0.450, p < 0.001). A low amniotic PP13 level significantly predicted the birth weight (r = -0.102, p < 0.05), the duration of pregnancy (r = -0.155, p < 0.05), and the fetal abdominal circumference (r = -0.098, p < 0.05). Conclusions: PP13 assayed in amniotic fluid might be a potential marker of fetal growth, and sHLA-G can be an adjunct modality reflecting placental sonographic parameters.

Fatty acids and their metabolites (resolvins) are altered in women with gestational diabetes mellitus (GDM).

The maternal fatty acid status plays a key role in influencing pregnancy outcomes. Omega-3 fatty acids are the precursors for E-series (RvE) and D-series resolvins (RvD) and possess anti-inflammatory properties. Pregnancy complications like gestational diabetes mellitus (GDM) are associated with excess maternal inflammation. This study reports the levels of maternal fatty acids across gestation in GDM and non-GDM women, placental fatty acids, resolvins and their association with the maternal fatty acid status. Pregnant women were recruited at 11-14 (V1) weeks and followed at 18-22 (V2) and 26-28 (V3) weeks and at delivery (V4). A total of 209 women who were diagnosed as GDM and 207 non-GDM women were included in this study. Fatty acids were estimated using gas chromatography. The protein levels of resolvins (RvE1, RvE2, RvD1 and RvD2) were measured using ELISA kits. Total PUFAs, eicosapentaenoic acid (EPA), omega-6 fatty acids, linoleic acid (LA) and arachidonic acid (AA) were lower, while saturated fatty acid (SFA) and alpha-linolenic acid (ALA) levels were higher in GDM women at 18-22 weeks. Placental AA was lower (p < 0.05) in women with GDM. Placental protein levels of RvE1, RvD1 and RvD2 were lower (p < 0.001 for all) in the GDM group. The maternal delta 5 desaturase index was positively associated, while erythrocyte omega-3 and omega-6 fatty acids were negatively associated with RvE2 at 11-14 weeks. Placental LA and ALA were positively associated with RvD1 and RvD2 (p < 0.05, for both), respectively. Our findings suggest that the maternal fatty acid status influences pro-resolving mediators which may lead to increased inflammation in GDM.

Smoking during pregnancy is associated with the placental proteome

Maternal smoking during pregnancy (MSDP) is a significant risk factor for the development of foetal, neonatal, and childhood morbidities. We hypothesized that infants exposed to MSDP have a distinct proteomic expression in their term placentas compared to infants without such an exposure. A total of 39 infants exposed (cord blood cotinine levels of >1 ng/mL) and 44 infants not exposed to MSDP were included in the study. Women with chronic disease, body mass index of > 30, or a history of uterine surgery were excluded. Total proteome abundance was analysed with quantitative mass spectrometry. For univariate analysis of differences in placental protein levels between groups, ANOVA with multiple testing corrections by the Benjamini–Hochberg method was used. For multivariate analysis, we used principal component analysis, partial least squares, lasso, random forest, and neural networks. The univariate analyses showed four differentially abundant proteins (PXDN, CYP1A1, GPR183, and KRT81) when heavy and moderate smoking groups were compared to non-smokers. With the help of machine learning, we found that an additional six proteins (SEPTIN3, CRAT, NAAA, CD248, CADM3, and ZNF648) were discriminants of MSDP. The placental abundance of these ten proteins together explained 74.1% of the variation in cord blood cotinine levels (p = 0.002). Infants exposed to MSDP showed differential abundance of proteins in term placentas. We report differential placental abundance of several proteins for the first time in the setting of MSDP. We believe that these findings supplement the current understanding of how MSDP affects the placental proteome.

Evaluation of maternal serum SERPINC1, E-selectin, P-selectin, RBP4 and PP13 levels in pregnancies complicated with preeclampsia

Objective The aim of this study is to investigate whether Serpin clade C (SERPINC1), E-selectin, P-selectin, Placental protein 13 (PP13), and Retinol-binding protein-4 (RBP4) levels in maternal serum were associated with the presence of preeclampsia and to compare them with uncomplicated pregnancies. Methods This prospective study included 40 women with preeclampsia and 40 healthy pregnant women. An enzyme-linked immunosorbent assay kit was used to measure serum SERPINC1, E-selectin, P-selectin, PP13, and RBP4 levels. Results The preeclampsia group had significantly higher E-selectin and P-selectin levels than the control group. PP13 and SERPINC1 levels were also significantly lower than the control group. There was no significant difference in RBP4 levels. The receiver operating characteristic curve revealed the best cutoff values for the following: E-selectin >19.2 ng/mL, with 87.5% sensitivity and 95% specificity; P-selectin >5.1 ng/mL, with 97.5% sensitivity and 100% specificity; PP13 ≤ 107.03 pg/mL, with 72.5% sensitivity and 77.5% specificity; and SERPINC1 ≤ 87.76 ng/mL, with 100% sensitivity and 97.5% specificity. Conclusion In this study, the endothelial dysfunction parameters SERPINC1, PP13, E-selectin, and P-selectin were found to be associated with preeclampsia. Endothelial dysfunction biomarkers in maternal non-serum body fluids may differ. More research is needed, especially to determine the relationship between SERPINC1 and preeclampsia.

Proteomic Approaches in the Study of Placenta of Pregnancy Complicated by Gestational Diabetes Mellitus.

Gestational diabetes mellitus (GDM), a glucose intolerance developing or first recognized during pregnancy, leads to a series of short- and long-term maternal and fetal complications, somehow related to placenta structural and functional changes. The focus and the objective of the present review are to discuss the results which can be obtained by different mass spectrometric approaches in the study of placenta protein profile. Thus, matrix-assisted laser desorption/ionization mass spectrometry (MALDI) has been applied on placenta omogenates before and after one-dimensional electrophoretic separation, followed by tryptic digestion. MALDI imaging was used for direct investigation on the placenta tissue (both maternal and fetal sides). The results showed that some differences among the absolute abundances of some proteins are present for placenta samples from GDM patients. The majority of investigations were carried out by two-dimensional electrophoresis (2DE) followed by LC-MS/MS or, directly by the label-free LC-MSE approach. It should be emphasized that all these techniques were showed differences in the protein expression between the placenta samples from healthy or GDM subjects. 2DE was also employed to separate and compare placental protein levels from GDM and the control groups: differentially expressed proteins between the two groups were identified by MALDI-TOF/TOF mass spectrometry and were further confirmed by Western blotting. The physiopathological significance of the obtained results are reported and discussed in this narrative review. The experimental data obtained until now show that the newest, mass spectrometric approaches can be considered a valid tool to investigate the possible changes of placenta in the presence of GDM.

Placental protein levels in maternal serum are associated with adverse pregnancy outcomes in nulliparous patients

The Eunice Kennedy Shriver National Institute of Child Health and Human Development Nulliparous Pregnancy Outcomes Study: Monitoring Mothers-to-Be was established to investigate the underlying causes and pathophysiological pathways associated with adverse pregnancy outcomes in nulliparous gravidas. This study aimed to study placental physiology and identify novel biomarkers concerning adverse pregnancy outcomes, including preterm birth (medically indicated and spontaneous), preeclampsia, small-for-gestational-age neonates, and stillbirth. We measured levels of placental proteins in the maternal circulation in the first 2 trimesters of pregnancy. Maternal serum samples were collected at 2 study visits (6-13 weeks and 16-21 weeks), and levels of 9 analytes were measured. The analytes we measured were vascular endothelial growth factor, placental growth factor, endoglin, soluble fms-like tyrosine kinase-1, A disintegrin and metalloproteinase domain-containing protein 12, pregnancy-associated plasma protein A, free beta-human chorionic gonadotropin, inhibin A, and alpha-fetoprotein. The primary outcome was preterm birth between 20 0/7 and 36 6/7 weeks of gestation. The secondary outcomes were spontaneous preterm births, medically indicated preterm births, preeclampsia, small-for-gestational-age neonates, and stillbirth. A total of 10,038 eligible gravidas were enrolled in the Nulliparous Pregnancy Outcomes Study: Monitoring Mothers-to-Be cohort, from which a nested case-control study was performed comparing 800 cases with preterm birth (466 spontaneous preterm births, 330 medically indicated preterm births, and 4 unclassified preterm births), 568 with preeclampsia, 406 with small-for-gestational-age birth, and 49 with stillbirth with 911 controls who delivered at term without complications. Although levels of each analyte generally differed between cases and controls at 1 or 2 visits, the odds ratios revealed a <2-fold difference between cases and controls in all comparisons. Receiver operating characteristic curves, generated to determine the relationship between analyte levels and preterm birth and the other adverse pregnancy outcomes, resulted in areas under the receiver operating characteristic curves that were relatively low (range, 0.50-0.64) for each analyte. Logistic regression modeling demonstrated that areas under the receiver operating characteristic curves for predicting adverse pregnancy outcomes were greater using baseline clinical characteristics and combinations of analytes than baseline characteristics alone, but areas under the receiver operating characteristic curves remained relatively low for each outcome (range, 0.65-0.78). We have found significant associations between maternal serum levels of analytes evaluated early in pregnancy and subsequent adverse pregnancy outcomes in nulliparous gravidas. However, the test characteristics for these analytes do not support their use as clinical biomarkers to predict adverse pregnancy outcomes, either alone or in combination with maternal clinical characteristics.

Placental mRNA and Protein Expression of VDR, CYP27B1 and CYP2R1 Genes Related to Vitamin D Metabolism in Preeclamptic Women

(1) Background: Considerable evidence indicates that the occurrence of preeclampsia (PE) is associated with a reduced vitamin D (VD) level. Several studies have found that VD deficiency is correlated with disturbed trophoblast invasion, reduced angiogenesis and increased vasoconstriction. Because the vitamin D receptor (VDR) and CYP27B1 and CYP2R1 hydrolases are strongly involved in VD metabolism, the goal of the present study was to evaluate their genes and proteins expression in the placentas from preeclamptic women. (2) Methods: Samples and clinical data were obtained from 100 Polish women (41 women with preeclampsia and 59 healthy pregnant controls). The whole PE group was divided into subgroups according to gestation week of pregnancy ending before and after 34 gestational weeks (early/late-onset preeclampsia (EOPE/LOPE)). However, finally, to reduce confounding by differences in gestational age, the EOPE group was excluded from the analysis of mRNA and protein placental expression, and we focus on the comparison between LOPE and control groups. The placental VDR, CYP27B1 and CYP2R1 mRNA expression was analyzed using RT-PCR, and placental protein levels were determined by ELISA assay. (3) Results. (3.1) Placental gene expression: Expression levels of both genes, CYP27B1 (1.17 vs. 1.05 in controls, p = 0.006) and CYP2R1 (2.01 vs. 1.89 in controls, p = 0.039), were significantly higher in preeclamptic placentas than in the control group. Interestingly, VDR expression was significantly lower in placentas from the PE group (1.15 vs. 1.20 in controls, p = 0.030). After dividing all preeclamptic women into subgroups only for the CYP27B1 gene, a significantly higher placental expression in the LOPE subgroup than the healthy controls was observed (padj = 0.038). (3.2) Placental protein expression: The results revealed that protein expression levels of CYP27B1 in the preeclamptic group were similar (5.32 vs. 5.23 in controls, p = 0.530). There was a significant difference in median VDR and CYP2R1 protein levels between studied groups (VDR: 2.56 vs. 3.32 in controls, p &lt; 0.001; CYP2R1: 1.32 vs. 1.43 in controls, p = 0.019). After stratification of preeclamptic women into subgroups, a significant difference was observed only in the VDR protein level. The medians in the LOPE subgroups were significantly lower compared to the healthy control group. In the whole study group, the placental VDR protein level was inversely correlated with systolic and diastolic blood pressure (all p &lt; 0.001), and positively correlated with gestational age (p &lt; 0.001) and infant birth weight (p = 0.014). (4) Conclusions: Lower mRNA and protein expression of VDR in preeclamptic placentas, and also VDR protein expression, could play a pivotal role in preeclampsia development. Additionally, the higher mRNA expression of both CYP27B1 and CYP2R1 hydrolase genes in placentas from preeclamptic women could indicate the compensatory role of these enzymes in preeclampsia etiology. Our results also indicate that placental VDR protein level could be one of the factors modulating blood pressure in pregnant women, as well as influencing gestational age and infant birth weight. Considering the importance of these findings, future studies are warranted.

Circulating heat shock protein 27 (HSPB1) levels in prediction of pre-eclampsia: A pilot study.

To determine whether circulating heat shock proteins HSP27/HSPB1 and HSP90α/HSPC1may be useful for early prediction of the occurrence of pre-eclampsia in asymptomatic women. We have measured by ELISA the levels of HSPB1, HSPC1, and placental protein 13 (PP13) in serum samples from 44 women in the first trimester (10-12weeks) and second trimester (17-20weeks) of pregnancy. Western blot and immunohistochemistry for HSPB1 and HSPC1 were performed. HSPB1serum levels were higher in women with pre-eclampsia than in normotensive pregnant women at the first and second trimester (P=0.003), whereas PP13levels decreased in women with pre-eclampsia only in the first trimester of gestation (P=0.021). We also observed higher HSPB1levels in patients with early-onset pre-eclampsia in the first and second trimester (P=0.014). This pilot study points out that circulating HSPB1levels in first and second trimester might be useful for predicting the occurrence of pre-eclampsia in asymptomatic women. Further validation studies are needed to finally establish this protein as a candidate predictive biomarker of pre-eclampsia.

Relevance of Placental protein 13 in combination with Xanthine oxidase activity andAsymmetric dimethylargininein early prediction of Preeclampsia with an evidence of LGALS13 gene promoter variant

Introduction and Aim:Preeclampsia is a pregnancy specific hypertensive disorder and its complications are associated with maternal morbidity and mortality. The intention of the study is to determine early placental and endothelial dysfunction markers in first and second trimesters of pregnancy for the prediction of preeclampsia.&#x0D; Materials and Methods:Study design was prospective which enrolled 268 pregnant women during their first ante-natal visit. Blood samples were analysed for Placental protein13 (PP13), Asymmetric dimethyl arginine (ADMA) by ELISA and Xanthine oxidase (XO) activity by colorimetric assay. On follow-up, 22 subjects developed preeclampsia, 12 had spontaneous abortions and the rest remained normotensive till delivery. PP13 gene, LGALS13 was sequenced in preeclampsia cases to know if any polymorphism was present.&#x0D; Results:Results were analysed by Wilcoxon Signed Ranks test know the level of statistical significance, correlation analysis by Spearman’s correlation and diagnostic performance of the measured parameters was assessed by receivers operating characteristic (ROC) curve. Pregnant women who developed Preeclampsia had low PP13 levels in first trimester and elevated ADMA in second trimester (p&lt;0.001). Gene sequencing of LGALS13 in preeclampsia showed -98C/A SNP at promoter region. ROC curve showed AUC for first trimester PP13 (0.932), second trimester PP13 (0.955) second trimester XO activity (0.967) and second trimester ADMA (1.000).&#x0D; Conclusion: The results suggest that oxidative stress, placental and endothelial dysfunction biomarkers would be helpful for early prediction of preeclampsia.

Intergenerational effects of the antioxidant Idebenone on the placentas of rats with gestational diabetes mellitus

Experimental models of maternal diabetes lead to the intrauterine programming of Gestational Diabetes Mellitus (GDM) in the offspring, together with an intrauterine proinflammatory environment, feto-placental metabolic alterations and fetal overgrowth. The aim of this work was to evaluate the effect of the mitochondrial antioxidant Idebenone given to F0 mild pregestational diabetic rats on the development of GDM in their F1 offspring and the intergenerational programming of a pro-oxidant/proinflammatory environment that affects the placentas of F2 fetuses. Control and mild pregestational diabetic female rats (F0) were mated with control males, and Idebenone or vehicle was administered to diabetic rats from day 1 of gestation to term. The F1 female offspring were mated with control males and maternal and fetal plasma samples were obtained for metabolic determinations at term. The F2 fetuses and placentas were weighed, and placental protein levels and peroxynitrite-induced damage (immunohistochemistry), mRNA levels (PCR), nitric oxide production (Griess reaction), and number of apoptotic cells (TUNEL) were evaluated. The F1 offspring of F0 diabetic rats (treated or not with Idebenone) developed GDM. The placentas of GDM rats showed a decrease in the mRNA levels of manganese superoxide dismutase and an increase in the production of nitric oxide, peroxynitrite-induced damage, and connective tissue growth factor levels, alterations that were prevented by the maternal Idebenone treatment in F0 rats. In conclusion, the maternal treatment with Idebenone in pregestational diabetic F0 rats ameliorates the pro-oxidant/proinflammatory environment that affects the placentas of F2 fetuses, although it does not prevent F1 rats from developing GDM.

Maternal Serum Placental Protein-13 Levels in the Prediction of Pregnancies with Abnormal Invasive Placentation.

To evaluate whether placental protein-13 (PP-13) measured in the serum of pregnant women could predict abnormal invasive placentation (AIP) detected by color Doppler ultrasound (US) and magnetic resonance imaging scan in addition to the routine US scan during the third trimester. The prospective case-control study included patients subdivided in 2 groups: 42 pregnant women with a singleton pregnancy at 28-32 weeks of gestation with only suspected AIP, and 32 healthy pregnant women. The serum PP-13 levels were measured in both groups using an enzyme-linked immunosorbent assay (ELISA) method and statistically compared. The cases of AIP were confirmed by placental histopathological examination and/or the uterus removed by hysterectomy after elective caesarean section. Serum PP-13 levels of pregnant women with AIP were significantly higher (p<0.001) than those of controls (650.32±387.33 vs. 231.43±94.33). Statistical analysis of maternal serum PP-13 levels above the threshold of 312 pg/ml (measured in the early third trimester) predicted AIP with 76.2% sensitivity and 75% specificity. Maternal serum PP-13 may have a role in the pathophysiology of AIP owing to its high serum value in the AIP group. The maternal serum dosage of PP-13 levels could improve pregnancy management in those patients suspected of having AIP.

Association between urinary placental protein 13 and soluble fms-like tyrosine kinase-1 in preeclamptic women in the third trimester of pregnancy

Background: Preeclampsia (PE) is diagnosed after 20 weeks of gestation. This multisystem disorder affects 2-7% of pregnant women. PE is a serious complication of pregnancy and one of the main causes of maternal and neonatal mortality and morbidity in the world. The inadequate placentation process results in a change in anti-angiogenic factors levels, such as placental protein 13 (PP13) and soluble fms-like tyrosine kinase 1 (sFlt-1). Objectives: To investigate the correlation between urinary PP13 and sFlt-1 in preeclamptic women in their third trimester of pregnancy. Methods: A case-control study was carried out from August 2018 till January 2019. Urine samples were collected from pregnant women at Al-Elweyia Hospital, Al-Hakeem Hospital, and Al- Imamain Al-Kadhimain Medical City (in Baghdad, Iraq). The patients’ group include fifty women with PE in the third trimester (25 mild and 25 severe). Fifty healthy pregnant women (at their third trimester of pregnancy) were studied as a control group. Results: The mean urinary PP13 levels were decreased in women with PE significantly (p≤ 0.05) (mild and severe) compared with healthy women (43.44±4.914 pg/ml, 33.34±1.863 pg/ml, and 51.84 ±2.60 pg/ml) respectively. Also, urinary sFlt-1 concentrations were decreased non-significantly (p&gt; 0.05) in women with PE (mild and severe) compared with healthy women (5.71±0.414 ng/ml, 5.31±0.38 ng/ml and 6.01±0.282 ng/ml), respectively. Conclusions: Urinary PP13 and sFlt-1 levels in the third trimester of pregnancy were significantly correlated with the severity of PE, and urinary levels of PP13 were found to be decreased significantly in patients with PE than in healthy pregnant women in the 3rd trimester of pregnancy.

Prediction of preeclampsia using first trimester placental protein 13 and uterine artery Doppler

Objective To determine the predictive value for preeclampsia by using serum placental protein 13 (PP13) levels and uterine artery pulsatility index (PI) in the first trimester. Methods This is a prospective observational study that was conducted in pregnant women with gestational age 11–13+6 weeks. Transabdominal uterine artery Doppler and serum PP13 level were performed at the first trimester aneuploidy screening visit. The predictive values of these tests were calculated. Results Data from 353 pregnant women were analyzed. Twenty-nine cases developed preeclampsia. The sensitivity, specificity, positive predictive value and negative predictive value of serum PP13 levels in predicting preeclampsia were 51.7, 65.7, 11.9, and 93.8%, respectively. The sensitivity, specificity, positive predictive value and negative predictive value of the uterine artery PI in predicting preeclampsia were 10.3, 95.7, 17.7, and 92.3%, respectively. When a combination of serum PP13 levels and uterine artery PI were used to predict preeclampsia, the sensitivity, specificity, positive predictive value and negative predictive value were 58.6, 62.9, 12.4 and 94.4%, respectively. Conclusion This study demonstrated that the combination of serum PP13 level and uterine artery Doppler in the first trimester was increased the sensitivity for predicting preeclampsia.

The Effects of Early-Onset Pre-Eclampsia on Placental Creatine Metabolism in the Third Trimester.

Creatine is a metabolite important for cellular energy homeostasis as it provides spatio-temporal adenosine triphosphate (ATP) buffering for cells with fluctuating energy demands. Here, we examined whether placental creatine metabolism was altered in cases of early-onset pre-eclampsia (PE), a condition known to cause placental metabolic dysfunction. We studied third trimester human placentae collected between 27–40 weeks’ gestation from women with early-onset PE (n = 20) and gestation-matched normotensive control pregnancies (n = 20). Placental total creatine and creatine precursor guanidinoacetate (GAA) content were measured. mRNA expression of the creatine synthesizing enzymes arginine:glycine aminotransferase (GATM) and guanidinoacetate methyltransferase (GAMT), the creatine transporter (SLC6A8), and the creatine kinases (mitochondrial CKMT1A & cytosolic BBCK) was assessed. Placental protein levels of arginine:glycine aminotransferase (AGAT), GAMT, CKMT1A and BBCK were also determined. Key findings; total creatine content of PE placentae was 38% higher than controls (p < 0.01). mRNA expression of GATM (p < 0.001), GAMT (p < 0.001), SLC6A8 (p = 0.021) and BBCK (p < 0.001) was also elevated in PE placentae. No differences in GAA content, nor protein levels of AGAT, GAMT, BBCK or CKMT1A were observed between cohorts. Advancing gestation and birth weight were associated with a down-regulation in placental GATM mRNA expression, and a reduction in GAA content, in control placentae. These relationships were absent in PE cases. Our results suggest PE placentae may have an ongoing reliance on the creatine kinase circuit for maintenance of cellular energetics with increased total creatine content and transcriptional changes to creatine synthesizing enzymes and the creatine transporter. Understanding the functional consequences of these changes warrants further investigation.

Prenatal vitamin D supplementation reduces blood pressure and improves placental angiogenesis in an animal model of preeclampsia.

Preeclampsia (PE), a pregnancy complication, is characterized by abnormal placental angiogenesis. The current study examines the effect of vitamin D deficiency/supplementation on pregnancy outcome and placental angiogenesis using an animal model of PE. Pregnant Wistar rats were divided into four groups: Control; PE; Vitamin D deficient with PE (VDD-PE) and Vitamin D supplemented with PE (VDS-PE). PE was induced by administering l-nitroarginine methyl ester (l-NAME) at the dose of 50 mg per kg body weight per day from day 14 to day 19 gestation in all the 4 groups. During the pre-pregnancy and pregnancy period, the rats from the Control and PE groups were fed a control diet, the VDD-PE group received a vitamin D deficient diet and the VDS-PE group received a vitamin D supplemented diet. Dams were sacrificed at d20 of gestation. l-NAME administration increased systolic as well as diastolic blood pressure in both PE and VDD-PE groups as compared to the control (p < 0.01). Vitamin D supplementation was beneficial in reducing the blood pressure. Vitamin D deficiency also lowered the placental protein levels of pro-angiogenic proteins VEGF and Flt-1 (p < 0.05 and p < 0.01, respectively), while the levels of these proteins in the VDS-PE group were similar to those in the control group. Vitamin D status did not influence the levels of PlGF and Hif1α. A low dose vitamin D supplementation given from pre-pregnancy and throughout pregnancy was beneficial in reducing the blood pressure and normalizing the placental levels of VEGF and Flt-1. This has implications for reducing the severity of preeclampsia.

Placental protein 13 and asymmetric dimethyl arginine for early assessment of preeclampsia

Objective: To evaluate serum levels of biomarkers as early indicators of onset of preeclampsia during pregnancy. Method: A nested case-control study was conducted among the pregnant women who visited for antenatal check-up at R.L. Jalappa Research Center, Kolar between August 2017 and May 2018. Serum levels of placental protein 13 (PP13), asymmetric dimethylarginine (ADMA), nitric oxide and caspase-3 which represents placentation and endothelial function were measured and compared between trimesters. Results and Discussion: There were 86 pregnant women enrolled at first trimester, the baseline values of women who developed preeclampsia were PP13 (477.54 ± 62.72 pg/ml), ADMA (21.06 ± 19.42 ng/ml), nitric oxide (3.71 ± 2.76 nmol/μL) and caspase-3 (6.15 ± 0.55 ng/ml). In the second trimester on follow up, the mean serum concentration of PP13 was 530.64 ± 69.10 pg/ml in preeclampsia versus 518.26 ± 49.33 pg/ml in normal pregnancy. The mean serum concentration of ADMA was 73.43 ± 30.95 ng/ml and nitric oxide 4.10 ± 2.70 nmol/μL in preeclampsia versus 21.06 ± 19.42 ng/ml and 4.85 ± 2.19 nmol/μL in normal pregnancy. The mean serum concentration of caspase-3 in preeclampsia was 22.65 ± 0.91 ng/ml versus normal pregnancy 18.45 ± 2.62 ng/ml were recorded. In preeclampsia, PP13 was positively correlated with nitric oxide and negatively correlated with ADMA. Conclusion: Decreased PP13, nitric oxide and elevated ADMA in first trimester and increased PP13, ADMA and decreased nitric oxide at second trimester reflects altered placentation and endothelial function.

285. Syncytiotrophoblast derived human placental microvesicles and exosomes carry placental protein-13 (PP13) and their level is decreased in preeclampsia

Background Extracellular vesicles (EVs) are communicating physio/pathological signals between organs. In preeclampsia (PE) – a life threatening pregnancy complication - there is an elevated shedding of EVs by the placental syncytiotrophoblast (STB-EVs) into the maternal circulation. Different STB-EVs subtypes carries each a large repertoire of proteins and RNAs to signal the maternal organs. Here we set to determine placental protein 13 (PP13) to identify if it is included in the STB-EVs’ cargo, whether it is located inside or outside, and if and how it change in PE given that maternal serum PP13 is reduced during the first trimester in patients who subsequently developed PE. Furthermore, PP13 causes expansion of the maternal uterine arteries to accommodate the increase blood flow in pregnancy, induces apoptosis of lymphocytes and enhances cytokines secretion related to immune functions. Methods Placentae were obtained at caesarean section deliveries of 20 PE and 19 controls. The STB-EVs were collected from the maternal side perfusate by the dual placental lobe perfusion, sequential centrifugation and filtration. Three population subtypes were isolated: STB microvesicles (STB-MVs), STB-Exosomes (STB-EXs) and total STB-EVs. Placental origin was assessed by Western blot and size distribution by Nanoparticle Tracking Analysis. PP13 was quantified by ELISA. Results All STB-EVs preparation had placental alkaline phosphatase (PLAP) biomarker. ALIX and CD9 were exclusively expressed in STB-EXs. PP13 was determined for the first time in all three STB-EVs preparations, and was expressed in both their inside and their surface. PP13 was significantly lower in PE compared to control in the total STB-EVs and STB-MVs, but not in STB-EXs. Conclusion At delivery there is a decreased in PP13 levels in STB-EVs derived from PE. Circulating PP13 is therefore either soluble or associated with STB-EVs, and each may deliver distinct PE associated signals. Funding MS was sponsored by Daniel Turnberg Fellowship, UK.

Reduced placental protein 13 (PP13) in placental derived syncytiotrophoblast extracellular vesicles in preeclampsia – A novel tool to study the impaired cargo transmission of the placenta to the maternal organs

IntroductionPlacental syncytiotrophoblast (STB) release extracellular vesicles (STB-EVs) that communicate physiological and pathological placental signals to the maternal organs. STB-EV release also increases in preeclampsia (PE). Here we explored the cargo of PP13 in STB-EVs from PE versus control placentas. MethodsPlacentae were harvested following cesarean section deliveries, and dual placental lobe perfusion was used to harvest STB-EV. Maternal side perfusate was centrifuged at 10,000 × g to yield the STB microvesicles, and then at 150,000 × g to yield STB exosomes. Total STB-EVs (tSTB-EVs) were collected using a one step 150,000 × g centrifugation. Placental origin and size distribution were assessed by Western blotting and Nanoparticle Tracking Analysis, respectively. PP13 expression was determined by Western blot and ELISA. ResultsPlacental alkaline phosphatase (PLAP; a STB specific marker) was present in all preparations. Total tSTB-EVs and STB-EXs also expressed the exosome markers such as the Apoptosis-Linked Gene 2-Interacting Protein X (Alix) and the cluster differentiation protein 9 (CD9). PP13 was localized to the outer surface and intra-vesicular compartments of all fractions. Surface to total PP13 ratios were ∼1:1 for all STB-EV preparations.In contrast to the previously reported higher circulating concentrations of soluble PP13 in PE, significantly lower levels of PP13, normalized to total vesicular protein, were observed in PE samples. PP13 reduction in all STB-EVs' sub-populations may be attributed to differences in gestational age (GA). A simple correction for GA suggested that PE may be an important influence. ConclusionsPP13 is located in and on all types of STB-EVs. Circulating PP13 may therefore be either soluble or associated with extracellular vesicles with different pathophysiological effects in the maternal circulation.

Early second trimester maternal serum markers in the prediction of gestational diabetes mellitus.

Aims/IntroductionTo determine whether maternal serum markers in the early second trimester are useful for prediction of gestational diabetes mellitus (GDM).Materials and methodsA total of 876 singleton pregnancies were recruited in the present study. Blood samples were collected during 16–20 gestational weeks. GDM women were diagnosed by an oral glucose tolerance test during 24–28 gestational weeks. A total of 56 women with GDM and 73 healthy pregnant women were selected. Maternal serum concentrations of placental protein 13 (PP13), pentraxin 3 (PTX3), soluble fms‐like tyrosine kinase‐1 (sFlt‐1), myostatin and follistatin (FST) were detected at 16–20 weeks’ gestation. All of these markers concentrations were expressed as multiples of the medians. The Mann–Whitney U‐test was used for comparison of the multiples of the medians of different concentrations of these five serum markers between the GDM group and the control group. Receiver operating characteristic curve analysis was applied to assess the sensitivity and specificity of significant serum markers from a Mann–Whitney U‐test comparison.ResultsCompared with healthy pregnancies, the serum levels of PP13, PTX3, sFlt‐1, myostatin and FST in the early second trimester were significantly increased in patients who had developed GDM late. In screening for GDM by PP13, PTX3, sFlt‐1, myostatin and FST, the detection rates were 92.3, 94.9, 94.9, 92.5 and 92.3%, respectively at 80% specificity. PTX3 and sFlt‐1 were the most sensitive markers.ConclusionsMaternal serum markers including PP13, PTX3, sFlt‐1, myostatin and FST increase in the early second trimester of women with GDM. These five markers, especially PTX3 and sFlt‐1, could have the value of prediction for those patients who would develop GDM in the late second trimester.