Pregnancy-related Disease Research Articles

Down-Regulation of Neuropathy Target Esterase in Preeclampsia Placenta Inhibits Human Trophoblast Cell Invasion via Modulating MMP-9 Levels

Background/Aims: Neuropathy target esterase (NTE, also known as neurotoxic esterase) is proven to deacylate phosphatidylcholine (PC) to glycerophosphocholine as a phospholipase B. Recently; studies showed that artificial phosphatidylserine/PC microvesicles can induce preeclampsia (PE)-like changes in pregnant mice. However, it is unclear whether NTE plays a key role in the pathology of PE, a pregnancy-related disease, which was characterized by deficient trophoblast invasion and reduced trophoblast-mediated remodeling of spiral arteries. The aim of this study was to investigate the expression pattern of NTE in the placenta from women with PE and normal pregnancy, and the molecular mechanism of NTE involved in the development of PE. Methods: NTE expression levels in placentas from 20 pregnant women with PE and 20 healthy pregnant women were detected using quantitative PCR and immunohistochemistry staining. The effect of NTE on trophoblast migration and invasion and the underlying mechanisms were examined in HTR-8/SVneo cell lines by transfection method. Results: NTE mRNA and protein expression levels were significantly decreased in preeclamptic placentas than normal control. Over-expression of NTE in HTR-8/SVneo cells significantly promoted trophoblast cells migration and invasion and was associated with increased MMP-9 levels. Conversely, shRNA-mediated down-regulation of NTE markedly inhibited the cell migration and invasion. In addition, silencing NTE reduced the MMP-9 activity and phosphorylated Erk1/2 and AKT levels. Conclusions: Our results suggest that the decreased NTE may contribute to the development of PE through impairing trophoblast invasion by down-regulating MMP-9 via the Erk1/2 and AKT signaling pathway.

Knockdown of Heparanase Suppresses Invasion of Human Trophoblasts by Activating p38 MAPK Signaling Pathway.

Preeclampsia is a pregnancy-related disease with increasing maternal and perinatal morbidity and mortality worldwide. Defective trophoblast invasion is considered to be a major factor in the pathophysiological mechanism of preeclampsia. Heparanase, the only endo-β-glucuronidase in mammalian cells, has been shown to be abnormally expressed in the placenta of preeclampsia patients in our previous study. The biological role and potential mechanism of heparanase in trophoblasts remain unclear. In the present study, stably transfected HTR8/SVneo cell lines with heparanase overexpression or knockdown were constructed. The effect of heparanase on cellular proliferation, apoptosis, invasion, tube formation, and potential pathways in trophoblasts was explored. Our results showed that overexpression of heparanase promoted proliferation and invasion. Knockdown of heparanase suppressed proliferation, invasion, and tube formation but induced apoptosis. These findings reveal that downregulation of heparanase may contribute to defective placentation and plays a crucial role in the pathogenesis of preeclampsia. Furthermore, increased activation of p38 MAPK in heparanase-knockdown HTR8/SVneo cell was shown by MAPK pathway phosphorylation array and Western blotting assay. After pretreatment with 3 specific p38 MAPK inhibitors (BMS582949, SB203580, or BIRB796), inadequate invasion in heparanase-knockdown HTR8/SVneo cell was rescued. That indicates that knockdown of heparanase decreases HTR8/SVneo cell invasion through excessive activation of the p38 MAPK signaling pathway. Our study suggests that heparanase can be a potential predictive biomarker for preeclampsia at an early stage of pregnancy and represents a promising therapeutic target for the treatment of preeclampsia.

322: Effects of selective reduced uterine perfusion pressure on organs commonly affected by preeclampsia

Selective reduced uterine perfusion pressure (SRUPP) has been used as a model of pregnancy related hypertensive disease in rats. In this study, we aimed to assess the effects of the SRUPP model on the weight of organs commonly affected by hypertensive disorders. 30 timed pregnant, Sprague-Dawley rats were divided into two groups, the SRUPP (n=30) or the control, sham surgery group (n=30). On gestational day 13 the rats underwent mean arterial pressure (MAP) analysis by tail cuff plethysmography. On gestational day 14, rats were anesthetized with 2% isoflurane and the abdomen entered through a midline vertical incision. Fetuses were counted, uterine and utero-ovarian arteries were identified and surgically isolated from the surrounding adipose tissue. In the SRUPP cohort, silver clips (0.102mm internal diameter) were placed on the aforementioned vessels bilaterally while the Sham surgery group vessels were solely isolated. The rats were returned to their home cages after surgery until gestational day 20 when MAP was again obtained. On day 21 the rats were terminally anesthetized with isoflurane. A second laparotomy was performed and organs of interest were harvested. Heart, kidney, mean fetal, and mean placental weights were obtained. Results are expressed in mean and standard error mean. Independent sample t-test was used for the statistical analysis. A two-sided p < 0.05 indicated a statistical significance. The heart weight was significantly greater in the SRUPP group when compared with controls (802.13 ± 32.804 vs 701.09 ± 19.548 mg; p= .012). There was no significant difference in the renal, mean fetal, or placental weight of SRUPP vs Control (See Table 1). SRUPP appears to be associated with a significant increase in heart weight. These changes suggest that in SRUPP the greater heart weight is a consequence of myocardial hypertrophy due to hypertension.

Prevalence and Risk Factor for Poor Pregnancy Outcome among Married Women, in Fafan Zone, Somali Region, Eastern Ethiopia

Introduction: Worldwide, over three hundred fifty thousand women of childbearing age die every year due to complications of pregnancy. Poor pregnancy outcomes are higher in poor countries as compared to developed countries. Objective: To asses prevalence and risk factor for poor pregnancy outcomes among married women, in Fafan zone, Somali regional state, eastern Ethiopia. Methodology: To conduct the study, community based retrospective cross-sectional study design was employed and systematic random sampling was used. Data was entered and analyzed by using SPSS version 21. The result of the study revealed that, 10.9% and 9.3% of the women had history of a still birth and abortion respectively in their reproductive life. Result: Overall, half of studied women (51.9.0%) fall under category of poor pregnancy outcome. Approximately one third (32%) of women had at least one type of chronic medical pregnancy related medical disease during pregnancy. For majority women (90.4%) included in the study FGM had been conducted. Another significant proportion of women (27%) were overweight (25.00–29.99). About, one third (34.7%) of respondents reported that they got pregnant at age less than 18 years. Being Somali in ethnicity (AOR=6.601; 95% CI 2.890-15.078), history of FGM (AOR=3.579; 95% CI 2.118-6.050) and pregnancy at ageless than18 years (AOR=2.4941; 95% CI, 0.753-3.547) were independent predictors for poor pregnancy outcome among married women. Conclusion: In conclusion, the study showed high prevalence of poor maternal and fetal pregnancy outcome and risk factor among studied women. Concerned bodies were recommended to focus on identification of potential risk factor, and intervene through preconception care before pregnancy.

Endothelial and kidney function in women with a history of preeclampsia and healthy parous controls: A case control study

IntroductionPreeclampsia (PE) is a pregnancy related endothelial disease characterized by hypertension and albuminuria. Postpartum endothelial dysfunction often persists in these women. We postulate that in women with a history of PE reduced endothelial dependent vasodilation coincides with attenuated kidney function, as both reflect endothelial dysfunction. MethodsWe assessed endothelial and kidney function in women with a history of PE (n=79) and uncomplicated pregnancies (n=49) at least 4years postpartum. Women with hypertension, diabetes or kidney disease prior to pregnancy were excluded. Brachial artery flow mediated dilatation (FMD) was measured and analysed by a custom designed edge-detection and wall-tracking software. We measured albumin and creatinine levels in a 24-h urine sample and calculated glomerular filtration rate (GFR) by CKD-EPI. ResultsWomen with a history of PE had lower FMD but comparable GFR and albumin creatinine ratio (ACR) compared with controls. Independent of obstetric history, in both controls and women with a history of PE respectively, GFR (r=0.19, p=0.17 and r=0.12, p=0.29) and albumin creatinine ratio (r=0.07, p=0.62 and r=0.06 p=0.57) did not correlate with FMD. ConclusionAt least 4years after pregnancy, women with a history of PE demonstrated decreased flow mediated dilatation when compared to healthy parous controls. In this study, decreased flow mediated dilation however did not coincide with decreased kidney function.

A simple pregnancy-related kidney disease screening method: random proteinuria testing.

A simple pregnancy-related kidney disease screening method: random proteinuria testing.

MicroRNAs: New Players in the Pathobiology of Preeclampsia.

Our understanding of how microRNAs (miRNAs) regulate gene networks and affect different molecular pathways leading to various human pathologies has significantly improved over the years. In contrary, the role of miRNAs in pregnancy-related hypertensive disorders such as preeclampsia (PE) is only beginning to emerge. Recent papers highlight that adverse pregnancy outcomes are associated with aberrant expression of several miRNAs. Presently, efforts are underway to determine the biologic function of these placental miRNAs which can shed light on their contribution to these pregnancy-related disease conditions. The discovery that miRNAs are stable in circulation coupled with the fact that the placenta is capable of releasing them to the circulation in exosomes generates a lot of enthusiasm to use them as biomarkers. In this review, we will summarize the recent findings of our understanding of miRNA regulation in relation to PE, a hypertensive disorder of pregnancy. Particular emphasis will be given to the role of key miRNA molecules such as miR-210 and miR-155 that are known to be consistently dysregulated in women with PE.

Whole-exome sequencing reveals genetic variants in ERC1 and KCNG4 associated with complete hydatidiform mole in Chinese Han women.

Complete hydatidiform mole (CHM) is a rare pregnancy-related disease with invasive potential. The genetics underlying the sporadic form of CHM have not been addressed previously, but maternal genetic variants may be involved in biparental CHM. We performed whole-exome sequencing of 51 patients with CHM and 47 healthy women to identify genetic variants associated with CHM. In addition, candidate variants were analyzed using single base extension and Matrix Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry in 199 CHM patients and 400 healthy controls. We validated candidate variants using Sanger sequencing in 250 cases and 652 controls, including 205 new controls. Two single nucleotide polymorphisms, c.G48C(p.Q16H) inERC1 and c.G1114A(p.G372S) in KCNG4, were associated with an increased risk of CHM (p<0.05). These variants may contribute to the pathogenesis of CHM and could be used to screen pregnant women for this genetic abnormality.

‘The obesity paradox’: a reconsideration of obesity and the risk of preterm birth

The association between obesity and spontaneous preterm births (sPTBs) has been shown to be influenced by obesity-attendant comorbidities. Our objective was to better understand the complex relationship of obesity and its attendant comorbidities with sPTBs. A retrospective analysis utilizing maternally linked hospital and birth certificate records of 2 049 196 singleton California deliveries from 2007 to 2011. Adjusted relative risks (aRRs) for sPTBs were estimated using multivariate Poisson regression modeling. Obese women had higher aRRs for sPTBs than their normal body mass index (BMI) controls. aRRs (95% confidence interval) increased with increasing BMI category: Obese I=1.10 (1.08 to 1.12); Obese II=1.15 (1.12 to 1.18); and Obese III=1.26 (1.22 to 1.30). When comparing only obese women without comorbidities to their normal BMI controls, aRRs reversed, that is, obese women had lower aRRs of sPTBs: Obese I=0.96 (0.94 to 0.98), Obese II=0.95 (0.91 to 0.98); and Obese III=0.98 (0.94 to 1.03). This same reversal of aRR direction was also observed among women with comorbidities: 0.92 (0.89 to 0.96); 0.89 (0.85 to 0.93); and 0.89 (0.85 to 0.93), respectively. Increasing BMI increased the aRRs for sPTBs among patients with gestational diabetes (P<0.05), while decreasing the risk among patients with chronic hypertension and pregnancy-related hypertensive disease (P<0.05). The obesity and preterm birth paradox is an example of what has been described as 'Simpson's Paradox'. Unmeasured confounding factors mediated by comorbidities may explain the observed protective effect of obesity upon conditioning on the presence or absence of comorbidities and thus resolve the paradox.

P 19 Alpha-1 microglobulin as a potential therapeutic candidate for the treatment of high blood pressure and oxidative stress in the STOX1 preeclampsia-model

Introduction Preeclampsia (PE) is a serious pregnancy-related disease with hypertension and proteinuria manifesting after 20 weeks of gestation. The worldwide prevalence is 3–8%, making it the most common cause of maternal and fetal mortality. PE lacks an effective therapy, and the only cure is delivery. We have previously shown that exogenously administered alpha-1-microglobulin (A1M) alleviates PE-like symptoms and the effects of oxidative stress in a dual-placenta perfusion model as well as in pregnant animal PE-models. Objectives A1M was evaluated as a therapeutic agent in the STOX1 PE-model. Materials and methods The STOX1 transgene mouse PE-model is one of the most complete PE-models available, since it reflects both stages of the disease and exhibit symptoms that mimic human PE. STOX1 is a transcription factor associated with PE that modulates proliferation and migration of trophoblasts. Wild type females are mated to STOX1-transgenic males, resulting in transgene-expressing fetuses and thereby restricting the expression to the utero-placental unit and typical PE symptoms in the female. Results One group of pregnant females received A1M injections i.p. every second day from day 6 of pregnancy until day 17, while another group received buffer. A1M significantly reduced the hypertension during gestation, especially at mid-gestation. It significantly reduced the level of hypoxia as well as the level of nitrative stress in the placenta. Also, A1M treatment reduced cellular damage in placenta and kidney tissue, such as disruption of plasma membranes, ER structures and mitochondria. However, A1M treatment had no significant effect on increased plasma levels of anti-angiogenic factors or the increased heart weight observed in the STOX1 PE-model. A1M was well tolerated with no side effects at the given dose. Conclusion A1M was able to alleviate PE symptoms observed in the STOX1 PE-model.

Abstract 2462: Whole-exome sequencing reveals genetic variants in ERC1 and KCNG4 in complete hydatidiform moles from Chinese Han women

Abstract Complete hydatidiform mole (CHM) is an uncommon pregnancy-related disease with an invasive potential. The genetic background of the sporadic CHM has not been addressed previously despite the possible mechanisms of maternal genetic variants to the development of this disease with biparental origin. We performed the whole-exome sequencing analysis on 51 CHM patients and 47 normal healthy women, then the second screening of the probable mutations by mass spectrometry in 199 CHM patients and 400 normal healthy women. Finally the candidate genetic polymorphisms were validated by direct Sanger sequencing in 247 cases and 599 controls with another 205 new controls. We eventually found that two SNPs c.G48C(p.Q16H) in ERC1 and c.G1114A(p.G372S) in KCNG4, were associated with an increased risk for CHM (p&amp;lt;0.05). These genetic variants will facilitate our understanding on the pathogenesis of CHM and related tumors as well as the oogenesis and embryonic implantation. They will provide beneficial information for the pregnant women both in physiology and psychology. Further multiple-disciplinary collaborations should be encouraged to clarify the accurate pathogenesis of CHM. Citation Format: Yan Yu, Bingjian Lu, Weiguo Lu, Xinyu Wang, Pengyuan Liu, Yan Lu, Xiaodong Cheng, Xing Xie. Whole-exome sequencing reveals genetic variants in ERC1 and KCNG4 in complete hydatidiform moles from Chinese Han women [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2462. doi:10.1158/1538-7445.AM2017-2462

Urinary metabolomic analysis of intrahepatic cholestasis of pregnancy based on high performance liquid chromatography/mass spectrometry

BackgroundIntrahepatic cholestasis of pregnancy (ICP), a pregnancy-related liver disease, leads to complications for both mothers and fetuses. Metabolomic approach has been applied to maternal-fetal medicine. The global metabolomic alterations that are specific in ICP as yet have not been investigated. MethodsBased on high performance liquid chromatography/hybrid quadrupole time-of-flight (HPLC/Q–TOF) mass spectrometry, the untargeted metabolomics was used to analyze the changes of urinary metabolites between ICP group and the control group. ResultsOne hundred nine variables in positive model and 119 variables in negative model were significantly different (p<0.05) between the ICP group and the control group, with the VIP (variable importance in the project) score>1 by the orthogonal partial least squares discriminant analysis (OPLS-DA). 14 metabolites in positive model and 18 metabolites in negative model were selected and identified based on HMDB (human metabolome database). Most of these metabolites were involved in bile acids biosynthesis and metabolism, hormone metabolism and lipid metabolism. A metabolite panel (MG (22:5), LysoPE (22:5), l-homocysteine sulfonic acid, glycocholic acid and chenodeoxycholic acid 3-sulfate) was contrusted by the binary logistic regression analysis with high diagnostic accuracy for ICP. The area under the receiver operating characteristic curve was 0.988 with the sensitivity of 90.0% and specificity of 93.3%. ConclusionsUrinary metabolites allow for the discrimination of ICP from the controls by orthogonal partial least squares discriminant analysis. Therefore, these findings may provide deep insights for the etiopathogenesis of ICP. Moreover, the maternal urinary metabolite panel has the potential to be used as non-invasive biomarkers for the diagnosis of ICP.

Age-, sex-, and diagnosis-specific incidence rate of medically certified long-term sick leave among private sector employees: The Japan Epidemiology Collaboration on Occupational Health (J-ECOH) study.

BackgroundLong-term sick-leave is a major public health problem, but data on its incidence in Japan are scarce. We aimed to present reference data for long-term sick-leave among private sector employees in Japan.MethodsThe study population comprised employees of 12 companies that participated in the Japan Epidemiology Collaboration on Occupational Health Study. Details on medically certified sick-leave lasting ≥30 days were collected from each company. Age- and sex-specific incidence rate of sick-leave was calculated for the period of April 2012 to March 2014.ResultsA total of 1422 spells in men and 289 in women occurred during 162,989 and 30,645 person-years of observation, respectively. The three leading causes of sick-leave (percentage of total spells) were mental disorders (52%), neoplasms (12%), and injury (8%) for men; and mental disorders (35%), neoplasms (20%), and pregnancy-related disease (14%) for women. Incidence rate of sick-leave due to mental disorders was relatively high among men in their 20s–40s but tended to decrease with age among women. Incidence rate of sick-leave due to neoplasms started to increase after age 50 in men and after age 40 in women, making neoplasms the leading cause of sick-leave after age 50 for women and after age 60 for men and the second leading cause after age 40 for women and after age 50 for men. Pregnancy-related disease was the second leading cause of sick-leave among women aged 20–39 years.ConclusionsThese results suggest that mental disorder, neoplasms, and pregnancy-related disease are the major causes of long-term sick-leave among private sector employees in Japan.

Evaluation of Admission Indications, Clinical Characteristics and Outcomes of Obstetric Patients Admitted to the Intensive Care Unit of a Teaching Hospital Center: A Five-Year Retrospective Review.

BackgroundCare of obstetric patients has always been a challenge for critical care physicians, because in addition to their complex pregnancy-related disease, fetal viability is considered.ObjectivesThe aim of this study was to review the admission indications, clinical characteristics and outcomes of obstetric patients, admitted to the intensive care unit of Alzzahra teaching hospital affiliated to Guilan University of Medical Sciences, Rasht, Iran.MethodsThis retrospective cohort study was conducted on pregnant /post-partum (up to 6 weeks) patients admitted to the ICU over a 5-year period from April 2009 to April, 2014.ResultsData from 1019 subjects were analyzed. Overall, 90.1% of the patients were admitted in the postpartum period. The most common indications for admission were pregnancy related hypertensive disorders (27.5%) and obstetric hemorrhage (13.5%). Epilepsy (5.4%) and cardiac disease (5.2%) were the most common non-obstetric indications.ConclusionsPregnancy-related hypertensive disorders and obstetric hemorrhage were the main reasons for admission, and epilepsy and cardiac disease were the most common non-obstetric indications. Efforts must be concentrated on increasing antenatal care.

A study of newborn on prelacteal feeds

Background: Infants are often prescribed prelacteal feeds after birth. We studied 50 such cases. Most prescriptions were written on day one. Maternal problems were the commonest reasons cited. However maternal and neonatal diseases severe enough to affect lactation are rarely encountered. On counselling breastfeeding was initiated. The seeds of lactation failure are sown on day one of life. The best chance of preventing it is on day one.Methods: 50 newborn babies born between August 2015 and July 2016 in Ashwini Rural Medical College and Hospital, Kumbhari, Solapur, Maharashtra, India and who were prescribed prelacteal feeds were included in the study. Approval from ethical committee was taken. Results: Most mothers did not have any medical or pregnancy related diseases. Antenatal counselling was not done in 18 (36 %) mothers. In 48 (96%) cases prelacteal feeds were initiated on day one. Maternal problems 19 (38%) were the commonest reasons cited for prelacteal feeds. In majority of cases 45 (90 %) there was no neonatal reason for prelacteal feeds. 22 (44 %) mothers initiated breastfeeding on recovery from their medical problems. In 17 (34%) cases, counselling was done and was effective.Conclusions: Prelacteal feeds are commonly prescribed in postnatal wards. Maternal problems were the commonest reasons cited though most of the mothers did not have medical or pregnancy related disease. Proper counselling was effective in convincing mothers. Maternal and neonatal diseases severe enough to affect lactation are rarely encountered. Antenatal counselling is an effective way to initiate breastfeeding on day one.

Lnc-DC mediates the over-maturation of decidual dendritic cells and induces the increase in Th1 cells in preeclampsia.

Preeclampsia is a severe pregnancy-related disease that involves an imbalance of the immune reaction, which is dominated by T helper1 (Th1) cells; however, the mechanism is unclear. We used flow cytometry, RT-PCR and western blotting to detect Th1 cells in the blood and mature dendritic cells, lnc-DC (a long non-coding RNA that is specifically expressed in DCs and can mediate dendritic cell maturation by phosphorylating STAT3) and p-STAT3 (phosphorylated transducer and activator of transcription 3) in the deciduas of normal pregnancies and preeclampsia patients. We found that the expression of lnc-DC and p-STAT3 was increased in the deciduas of preeclampsia patients. The proportion of Th1 cells and mature dendritic cells was significantly higher in the preeclampsia patients. These results suggested the hypothesis that the overexpression of lnc-DCs induces the over-maturation of decidual dendritic cells in preeclampsia patients and leads to an increase in Th1 cells.

Intrahepatic cholestasis of pregnancy and fetal outcomes. Mini review

Intrahepatic cholestasis of pregnancy (ICP), a pregnancy-related liver disease, leads to complications for both mother and fetus. It is the most common liver disease in pregnant women. The incidence of ICP has varied widely in different reports (ranging from 0.1 to 15.6%), for reasons that are incompletely understood. Hormonal, immunologic, genetic, environmental and alimentary factors have been implicated in the etiology of ICP. Although a clear correlation between elevated levels of maternal serum bile acids and deficient fetal outcome has been established in clinical practice, the underlying mechanisms remain elusive. Here we present a mini literature review about intrahepatic cholestasis of pregnancy and fetal outcomes.

Quantitative evaluation of longitudinal strain in layer-specific myocardium during normal pregnancy in China.

BackgroundThe myocardial wall of the left ventricle is a complex, multilayered structure and is not homogenous. The aim of this study was to determine longitudinal strain (LS) in the three myocardial layers in normal pregnant women according to gestation proceedings.MethodsThe advanced two-dimensional speckle tracking echocardiography (2D STE) was performed on 62 women during each pregnancy trimester and 6 to 9 weeks after delivery, while 30 age-matched, healthy, nonpregnant women served as controls. LS on endocardial, mid-myocardial and epicardial layers at 18 cardiac segments were measured.ResultsAs gestation proceeded, all of layer-specific LS and global LS progressively decreased, which subsequently recovered postpartum (P < 0.05), and the LS gradient between inner and outer myocardium became greater, which reached its maximum in the late pregnancy. Peak systolic LS was the highest at endocardium and the lowest at epicardium, while the highest at the apical level and the lowest at the base (P < 0.05). In the early pregnancy and postpartum, LS at basal level was homogenous, meanwhile layer-specific LS showed significant differences at mid-ventricular and apical level throughout the progress of normal pregnancy (P < 0.05).ConclusionsUsing 2D STE, three-layer assessment of LS can be performed in pregnant women and shall give us new insights into the quantitative analysis of global and regional LV function during pregnancy. Future studies on the detection of pregnancy related heart disease would require these parameters as reference values for each time point of a normal pregnancy.

Dysregulated circulating miRNAs in preeclampsia.

Preeclampsia (PE) is a pregnancy-related disease with potentially severe consequences with respect to foeto-maternal morbidity and mortality. However, the molecular pathogenesis of PE remains largely unknown. Recent reports have shown that microRNAs (miRNAs or miRs) may play important roles in the development of PE. Analysing the miRNAs in sera from preeclamptic women may improve our understanding of the pathophysiological mechanisms of the disease. The aim of this retrospective study was to identify whether circulating miRNAs were differentially expressed in PE patients compared with controls. Serum samples from 23 women who developed PE were compared with samples from 44 pregnant controls. Seventeen circulating miRNAs previously described in PE were chosen for evaluation of their expression by reverse transcription quantitative polymerase chain reaction (RT-qPCR). In the maternal serum, the miR-210-3p, miR-210-5p, miR-1233-3p, and miR-574-5p levels were found to be significantly higher in the PE patients than in the controls (P<0.05). Using a logistic regression model, we evaluated the discriminant power of those differentially expressed miRNAs, and the combination of miR-210-5p and miR-574-5p yielded an area under the curve of 0.7223 for discriminating PE patients from the controls. In conclusion, the fact that four circulating miRNAs (miR-210-3p, miR-210-5p, miR-1233-3p, and miR-574-5p) were differentially expressed in the sera of women who developed PE compared with controls confirms the possible pathophysiological role of miRNAs in PE.

The molecular relationship between antigenic domains and epitopes on hCG.

Antigenic domains are defined to contain a limited number of neighboring epitopes recognized by antibodies (Abs) but their molecular relationship remains rather elusive. We thoroughly analyzed the antigenic surface of the important pregnancy and tumor marker human chorionic gonadotropin (hCG), a cystine knot (ck) growth factor, and set antigenic domains and epitopes in molecular relationships to each other. Antigenic domains on hCG, its free hCGα and hCGβ subunits are dependent on appropriate inherent molecular features such as molecular accessibility and protrusion indices that determine bulging structures accessible to Abs. The banana-shaped intact hCG comprises ∼7500Å(2) of antigenic surface with minimally five antigenic domains that encompass a continuum of overlapping non-linear composite epitopes, not taking into account the C-terminal peptide extension of hCGβ (hCGβCTP). Epitopes within an antigenic domain are defined by specific Abs, that bury nearly 1000Å(2) of surface accessible area on the antigen and recognize a few up to 15 amino acid (aa) residues, whereby between 2 and 5 of these provide the essential binding energy. Variability in Ab binding modes to the contact aa residues are responsible for the variation in affinity and intra- and inter-species specificity, e.g. cross-reactions with luteinizing hormone (LH). Each genetically distinct fragment antigen binding (Fab) defines its own epitope. Consequently, recognition of the same epitope by different Abs is only possible in cases of genetically identical sequences of its binding sites. Due to combinatorial V(D)J gene segment variability of heavy and light chains, Abs defining numerous epitopes within an antigenic domain can be generated by different individuals and species. Far more than hundred Abs against the immuno-dominant antigenic domains of either subunit at both ends of the hCG-molecule, the tips of peptide loops one and three (Ł1+3) protruding from the central ck, encompassing hCGβŁ1+3 (aa 20-25+64+68-81) and hCGαŁ1 (aa 13-22; Pro16, Phe17, Phe18) plus hCGαŁ3 (Met71, Phe74), respectively, have been identified in the two "ISOBM Tissue Differentiation-7 Workshops on hCG and Related Molecules" and in other studies. These Abs recognize distinct but overlapping epitopes with slightly different specificity profiles and affinities. Heterodimeric-specific epitopes involve neighboring αŁ1 plus βŁ2 (hCGβ44/45 and 47/48). Diagnostically important Abs recognize the middle of the molecule, the ck (aa Arg10, Arg60 and possibly Gln89) and the linear hCGβCTP "tail" (aa 135-145; Asp139, Pro144, Gln145), respectively. Identification of antigenic domains and of specific epitopes is essential for harmonization of Abs in methods that are used for reliable and robust hCG measurements for the management of pregnancy, pregnancy-related disease and tumors.