Beta Subunit Of Chorionic Gonadotropin Research Articles

Evaluation and suggestions for improving the FIGO 2000 staging criteria for gestational trophoblastic neoplasia: A ten-year review of 1420 patients

BackgroundTo re-evaluate the efficacy of the prognostic factors currently employed in the treatment of malignant gestational trophoblastic neoplasia. MethodsClinical data from the Gestational Trophoblastic Disease (GTD) Center at Peking Union Medical Hospital (PUMCH) collected between January 2002 and December 2013 were retrospectively analyzed. Univariate and multivariate analyses of prognostic factors were performed using the Cox proportional hazards model. A new hazard ratio (HR)-based prognostic scoring scale was established and compared with the original scoring system. ResultsIn total, 1420 cases were included in the study (median follow-up=40months, overall complete remission (CR) rate=95.8%, relapse rate=7.1%, mortality rate=5.5%, median disease-free survival (DFS)=36months). Low-risk (0–6 points) and high-risk (≥6 points) patients exhibited CR rates of 99.8% (915/917) and 88.5% (445/503) and mortality rates of 0.3% and 15.1% (P<0.001), respectively. Univariate and multivariate analyses showed that age, pretreatment serum levels of human chorionic gonadotropin beta-subunit (β-hCG) and maximum tumor diameter were not independent prognostic risk factors. Antecedent pregnancy, the interval from the index pregnancy, the number of metastases and a history of failed chemotherapy treatments were independent prognostic risk factors. By modifying the scoring system based on the variables identified in a Cox analysis, we significantly increased the area under the receiver operating characteristics (ROC) curve. ConclusionThough effective, the accuracy of the International Federation of Gynecology and Obstetrics (FIGO) 2000 Trophoblastic Neoplasia Staging System requires improvement. Irrelevant prognostic factors should be removed, and the weights of other factors should be adjusted appropriately.

Selenoprotein K Mediates the Proliferation, Migration, and Invasion of Human Choriocarcinoma Cells by Negatively Regulating Human Chorionic Gonadotropin Expression via ERK, p38 MAPK, and Akt Signaling Pathway.

Selenoprotein K (SelK), a member of selenoprotein family, is identified as a single endoplasmic reticulum (ER) transmembrane protein. Although over-expression of SelK inhibits adherence and migration of human gastric cancer BGC-823 cells, the effects of SelK in human choriocarcinoma (CCA) are not well understood. In this study, the expression levels of SelK in three CCA cell lines, BeWo, JEG-3, and JAR, were examined. The effects of silencing or over-expressing SelK on expression of human chorionic gonadotropin beta subunit (β-hCG) were detected by western blotting. The results show that the protein level of β-hCG was reciprocally regulated by down- or up-regulation of SelK (*P<0.05; #P<0.05). The proliferative, migratory, and invasive capabilities of JEG-3 cells with reduced or over-expressed SelK were then tested using the cell counting kit-8 (CCK-8), wound healing, and transwell chamber assays. We found that these cellular activities were markedly increased by the loss of SelK in JEG-3 cells. Conversely, over-expressing SelK in JEG-3 cells suppressed these phenotypes. In addition, SelK expression after down- or up-regulation of β-hCG was also measured. Surprisingly, we found that level of SelK was affected by β-hCG (*P<0.05; #P<0.05). The proliferation, migration, and invasion were determined in JEG-3 cells after each over-expression and reduction of β-hCG. The results confirmed that β-hCG functions as a promoter of human choriocarcinoma. Furthermore, ERK/p38 MAPK and Akt signaling pathways were found to involve in these cellular functions. This work suggests that SelK may act as a tumor suppressor in human choriocarcinoma cells by negatively regulating β-hCG expression via ERK, p38 MAPK, and Akt signaling pathways. These findings revealed that selenoprotein K may serve as a novel target for human choriocarcinoma therapy in vitro.

Cell and genetic predictors of human blastocyst hatching success in assisted reproduction.

The aim was to identify cell and genetic predictors of human blastocyst hatching success in assisted reproduction programmes via a prospective case-control study. Blastocysts, donated by couples in assisted reproduction programmes were used. Hatching success assessment was performed after 144-146 h post-fertilization. The mRNA expression levels of cathepsin V (CTSV), GATA-binding protein 3 (GATA3) and human chorionic gonadotropin beta subunit 3, 5, 7 and 8 (CGB) genes were detected by quantitative real-time polymerase chain reaction. The odds ratio (OR) of hatching due to zona pellucida (ZP) thickness, oocyte and sperm quality, embryo quality and mRNA expression of CTSV, GATA3 and CGB genes in blastocysts was determined. From 62 blastocysts included in the study, 47 (75.8%) were unable to hatch spontaneously. The ZP thickening, and oocyte and sperm quality did not affect human blastocyst ability to hatch, except the combination of cytoplasmic and extracytoplasmic oocyte dysmorphisms (OR = 1.25; 95% confidence interval = 1.08, 1.45). Hatching-capable blastocysts had higher Gardner scale grade and mRNA expression of CTSV, GATA3 and CGB genes than hatching-incapable blastocysts. The human blastocyst hatching success depends on the blastocyst Gardner grade, but not on ZP and gamete quality. Blastocyst development was regulated by CTSV, GATA3 and CGB gene expression.

Effects of cigarette smoke extracts on cell cycle, cell migration and endocrine activity in human placental cells

Maternal smoking during pregnancy is known to be related to adverse pregnancy results associated with trophoblast proliferation and cell cycle progression. Moreover, many previous studies have shown that cigarette smoke is correlated with human chorionic gonadotropin beta (hCG-β) subunit produced from syncytiotrophoblasts during pregnancy. Thus, we further investigated whether cigarette smoke extract (CSE) affects the cell proliferation, migration and endocrine hormone activity of JEG-3 human placental cancer cells. JEG-3 cell proliferation was significantly reduced by all CSEs in a concentration-dependent manner. Moreover, CSEs decreased proliferating cell nuclear antigen (PCNA) levels in JEG-3 cells in Western blot. Increased migration or invasion ability of JEG-3 cells following CSE treatment was also confirmed by a scratch or fibronectin invasion assay in vitro. Additionally, protein levels of E-cadherin as an epithelial maker were down-regulated, while the mesenchymal markers N-cadherin, snail and slug were up-regulated in a time-dependent manner. The metastasis marker, cathepsin D, was also down-regulated by CSE. Finally, CSEs significantly reduced the expression of hCG-β protein in JEG-3 cells. Overall, these results indicate that exposure of placental cells to CSE deregulates the cell cycle by altering the expression of cell cycle-related proteins and stimulates cell metastatic ability by altering EMT markers and cathepsin D expression. CSE exposure may also decrease hCG-β production as an endocrine marker, implying that cigarette smoke has adverse effects during pregnancy.

Analysis of Down syndrome failed to be diagnosed after prenatal screening: A multicenter study.

To analyze the characters of Down syndrome (DS) who failed to be diagnosed after prenatal screening and hope to be able to improve the programs of prenatal screening and reduce the missed diagnosis of DS. In this multicenter study, we collected the missed cases from 3 prenatal diagnosis centers and analyzed their characters. A total of 126 DS babies failed to be diagnosed after prenatal screening. Their mothers accepted the prenatal screening in second trimester. We collected the mothers’ blood and detected the levels of alpha-fetoprotein (AFP) and the free beta subunit of human chorionic gonadotropin (fβhCG) by time-resolved fluoroimmunoassay. The values were also presented as multiples of the median (MoM) and determined the risk of carrying a fetus with DS by Wallace LifeCycle Elipse analysis software. Compared with normal control group, the level of fβhCG and hCG MoM were dramatically increased, while AFP and AFP MoM were decreased. The area under the receiver-operating-characteristic curve of trisomy 21 was 0.8387 for hCG-MoM and AFP-MoM testing. The sensitivity, specificity, positive predictive value, and negative predictive value were 84.6%, 74.8%, 75.4%, and 83.6%, respectively. Meanwhile, the prediction mode was “0.39957 + 1.90897∗HCG-MOM −3.32713∗AFP-MOM”. It was worthwhile noting that the risk of 65.9% DS missed diagnosis group were higher than 1/1000, 92.9% higher than 1/3000. However, 72.5% cases in normal control group were lower than 1/3000. Only 9.2% mothers would be higher than the value of risk in 1/1000. The prediction mode of hCG MoM and AFP MoM might be able to help us reduce the missed diagnosis. It is also necessary to adjust more reasonable range of noninvasive prenatal testing with further clinical researches.

Development of an inducible platform for intercellular protein delivery

A challenge to protein based therapies is the ability to produce biologically active proteins and their ensured delivery. Various approaches have been utilised including fusion of protein transduction domains with a protein or biomolecule of interest. A compounding issue is lack of specificity, efficiency and indeed whether the protein fusions are actually translocated into the cell and not merely an artefact of the fixation process. Here we present a novel platform, allowing the inducible export and uptake of a protein of interest. The system utilises a combination of the Tetracyline repressor system, combined with a fusion protein containing the N-terminal signal peptide from human chorionic gonadotropin beta-subunit, and a C-terminal poly-arginine domain for efficient uptake by target cells. This novel platform was validated using enhanced green fluorescent protein as the gene of interest. Doxycycline efficiently induced expression of the fusion protein. The human chorionic gonadotropin beta-subunit facilitated the export of the fusion protein into the cell culture media. Finally, the fusion protein was able to efficiently enter into neighbouring cells (target cells), mediated by the poly-arginine cell penetrating peptide. Importantly we have addressed the issue of whether the observed uptake is an artefact of the fixation process or indeed genuine translocation. In addition this platform provides a number of potential applications in diverse areas such as stem cell biology, immune therapy and cancer targeting therapies.

The Value of Serum Β-Subunit of Human Chorionic Gonadotropin Level in Prediction of Treatment Response to Methotrexate in Management of Ectopic Pregnancy; a Systematic Review and Meta-Analysis

Background: No consensus has been reached on prognostic value of serum concentration of β (beta) subunit of human chorionic gonadotropin (β-hCG) in treatment response to methotrexate in management of ectopic pregnancy. Therefore, the present study aimed to evaluate this subject through a systematic review and meta-analysis. Materials and Methods: An extensive literature search on online databases was performed. All studies performed on ectopic pregnancy patients treated by methotrexate from all age groups were included. After collecting data, random effect models were used to calculate the pooled standardized mean difference (SMD) of β-hCG level in treatment success and treatment failure groups. Finally, pooled performance screening characteristics of serum β-hCG level were assessed in different cut offs. Results: Finally, 51 articles were included in meta-analysis. Overall treatment success rate of methotrexate was 84% [95% confidence interval (CI): 84-85 percent]. A negative association was found between serum β-hCG level and the treatment response before intervention (SMD= -1.10, 95% CI: -1.39 to -0.88). In addition, pooled sensitivity, specificity, and prognostic odds ratio of β-hCG in the 2000 mIU/mL cut off were: 0.75 (0.65-0.82), 0.68 (0.58-0.82), and 6.0 (5.0-8.0), respectively. Conclusion: The present meta-analysis showed that serum β-hCG concentration before treatment could predict success of methotrexate in management of ectopic pregnancy.

The value of urea, creatinine, prolactin, and beta sub-unit of human chorionic gonadotropin of vaginal fluid in the diagnosis of premature preterm rupture of membranes in pregnancy.

Objective:To evaluate the effectiveness of urea, creatinine, prolactin, and the beta sub-unit of human chorionic gonadotropin (β-hCG) of vaginal fluid in the diagnosis premature preterm rupture of membranes (PROM).Materials and Methods:In this observational study, 160 pregnant women with gestational age of 28 to 40 weeks were divided into two equal groups: investigation (documented PROM) and control (intact membrane) groups. Five cubic centimeters of normal saline was poured into the vagina of all participants and the liquid was extracted after a few minutes using a syringe. The liquid was sent to a laboratory for examination. Data were analyzed using a t-test.Results:The volume of urea, creatinine, prolactin, and β-hCG was significantly different in the two groups (p<0.001). Based on receiver operating characteristic curve and cut-off point, sensitivity, specificity, positive and negative predictive values of β-hCG for detecting PROM were 87.5%, 86%, 86.4%, and 87.3%, respectively. Also, the same factors for urea in detecting PROM were 79.7%, 82.5%, 81.8%, and 80.4%, respectively. Creatinine had 74.6% sensitivity, 85% specificity, and 83% and 77.2% positive and negative predictive values for detecting PROM. Finally, prolactin had 87.5% sensitivity, 90% specificity, and 90% positive and 88% negative predictive values for detecting PROM.Conclusion:Prolactin and β-hCG have more diagnostic value than urea and creatinine in detecting PROM, and can be used in suspected cases.

Hysteroscopically-guided subchorionic methotrexate administration: A new technique for termination of cesarean scar pregnancy

Introduction: cesarean scar pregnancy (csP) is a rare but life-threatening condition, due to the risk of uterine rupture or for the development of placenta previa/accreta. the mainstay of management to spare fertility is early diagnosis and pregnancy termination. Although several conservative treatments have been experienced, no consensus on the best therapeutic approach is currently shared. the medical therapy for csP termination is based on systemic or local Methotrexate (MtX) administration. Hysteroscopy allows the precise visualization of placental implantation and it can be used to drive MtX injection selectively within the intervillous placental spaces. this procedure increases drug concentration delivered to the target tissue, potentially improving its therapeutic index. case report: A 36-year-old female with a history of cesarean section was admitted to our department with a diagnosis of viable 7th weeks csP. the patient gave her consent to pregnancy termination by a hysteroscopically-guided MtX administration within the cervico-isthmic placental implantation site, followed by its resectoscopic removal. In an office setting, 80 mg of MtX were injected through a needle adaptable to the 5-Fr operative channel of hysteroscope, Giancarlo Garuti1, Stefania Calabrese1, Lorenzo Quirino1, Marco Di Mario1 Affiliations: 1MD, Obstetrics and Gynecology Department, Public Hospital of Lodi, Via Savoia 1, 26900-Lodi, Italy. Corresponding Author: Giancarlo Garuti (MD), Obstetrics and Gynecology Department, Lodi Hospital, via Savoia n° 1, 26900-Lodi, Italy; E-mail: giancarlo.garuti@tiscali.it Received: 03 October 2015 Accepted: 02 December 2015 Published: 27 February 2016 under the chorionic membrane of placental implantation site. the duration of the procedure took three minutes and it resulted easy, painless and uneventful. An early embryo demise was obtained; the human chorionic gonadotropinbeta subunit (beta-hcG) serum trend showed a rapid pregnancy termination and no patient complaints were recorded. After 28 days, a resectoscopic removal of the gestational sac and placenta was safely accomplished. conclusion: selective hysteroscopic administration of MtX within intervillous spaces of the ectopic placental implantation is effective and it can be considered for csP termination.

Identification of novel transcription factors that may regulate transcription of the equine chorionic gonadotrophin beta subunit

Identification of novel transcription factors that may regulate transcription of the equine chorionic gonadotrophin beta subunit

Abstract 1047: A role for the free beta subunit of human chorionic gonadotropin in sensitivity of epithelial ovarian cancer cells to platinum-based chemotherapeutics

Abstract Introduction: The beta-subunit of human chorionic gonadotropin (hCG-β) is elevated in the serum of women with a range of malignant cancers, including epithelial ovarian cancer (EOC). Elevated levels of hCG-β have also been associated with poor response to radiotherapy and chemotherapy in a number of cancers. Women with EOC frequently respond to first line treatment with the platinum based chemotherapeutic drug cisplatin, but relapse with tumors developing chemoresistance. This study aimed to characterise the levels of hCG-β in EOC cell lines and to determine whether hCG-β has a role in the responsiveness of these cell lines to treatment with cisplatin. Methods: qRT-PCR and ELISA were used to determine endogenous levels of hCG-β in seven EOC cell lines: A2780, A2780cis (cisplatin resistant), HEY, SKOV-3, OVCAR-3, PEO-1 and OV202. hCG-β was down-regulated using siRNA in HEY and A2780cis cells. Sensitivity to cisplatin and taxol in these cells was measured using a clonogenic assay. Liquid chromatography-mass spectrometry using isobaric tags for relative and absolute quantitation (iTRAQ) was used to determine the mechanism(s) by which hCGβ may be involved in the sensitivity of EOC cells to cisplatin. Specifically, protein was harvested from A2780cis cells with and without hCGβ downregulation plus or minus treatment with cisplatin at IC50 levels (2μg/ml) for 24h. Protein extracts were then iTRAQ-labeled and global protein changes determined. Results: All cell lines expressed hCG-β transcript and protein. SKOV-3 and HEY cells secreted the highest levels, 14 ng/ml and 3 ng/ml, respectively. OV202, PEO-1, CaOV-3, A2780 and A2780cis cells expressed levels between 0.14-1.4 ng/ml. Down-regulation of hCG-β in HEY and A2780cis increased their sensitivity to cisplatin with the IC50 decreased up to 75% in knockdown compared to control cells. No change in sensitivity was observed with taxol treatment. Down-regulation of hCG-β also increased the sensitivity of these cells to the platinum drugs carboplatin and oxaliplatin. iTRAQ results showed differences in proteins associated with the cell cycle, death and survival pathways, as well as DNA replication, damage and repair. Candidate proteins are currently being validated by western blotting. Conclusions: These findings suggest that hCG-β may be involved in modulating the sensitivity of some EOCs to platinum based chemotherapy. Suppression of hCG-β may be a strategy to increase the responsiveness of primary EOCs to platinum-based chemotherapeutics. Citation Format: Snega M. Sinnappan, Robert C. Baxter, Deborah J. Marsh. A role for the free beta subunit of human chorionic gonadotropin in sensitivity of epithelial ovarian cancer cells to platinum-based chemotherapeutics. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1047. doi:10.1158/1538-7445.AM2015-1047

Comparing outcomes and costs between contingent and combined first-trimester screening strategies for Down's syndrome

ObjectiveTo compare a contingent strategy with a combined strategy for prenatal detection of Down's syndrome (DS) in terms of cost, outcomes and safety. Study designThe contingent strategy was based on a simulation, removing measurement of the free beta subunit of human chorionic gonadotropin (free βhCG) and calculating the DS risk retrospectively in 32,371 pregnant women who had been screened with the combined strategy in the first trimester. In the contingent strategy, a risk between 1:31 and 1:1000 in the first trimester indicated further testing in the second trimester (alpha-fetoprotein, inhibin A, unconjugated oestriol and free βhCG). The cut-off risk values for the contingent and combined strategies in the first trimester were 1:30 and 1:250, respectively, and the cut-off risk value for integrated screening in the second trimester was 1:250. Costs were compared in terms of avoided DS births, and the ratio of loss of healthy fetuses following invasive procedures per avoided DS birth was calculated. ResultsThe combined strategy had sensitivity of 40/44 (90.9%) and a false-positive rate of 2.8%. Corresponding values for the contingent strategy were 39/44 (88.6%) and 1.3%, respectively. Only 11% of pregnant women required tests in the second trimester, and the approximate cost reduction for each avoided DS birth was 5000€. The ratio of lost healthy fetuses following invasive procedures per avoided DS birth improved by up to 0.65. ConclusionThe contingent strategy has similar effectiveness to the combined strategy, but has lower costs and fewer invasive procedures.

Specific detection of type II human chorionic gonadotropin beta subunit produced by trophoblastic and neoplastic cells

BackgroundThe sequence of the beta-subunit of human chorionic gonadotropin (hCGβ) varies depending on whether hCGβ is encoded by type I or type II genes. Type II genes are upregulated in trophoblast and cancer but hCGβ can be detected in the serum of nonpregnant women and healthy individuals. We aimed to determine whether monoclonal antibody (mAb) FBT11-II specifically detects hCGβ encoded by type II genes (type II hCGβ). MethodsCompetitive inhibition assays with synthetic peptides, immunocytochemical and immunohistochemical studies, type II hCGβ dosing immunoassays and sequencing of CGB genes were performed. ResultsCompetitive inhibition assays determined that mAb FBT11-II recognizes the type II hCGβ derived peptide. CGB mRNA sequencing of JEG-3 (trophoblastic) and T24 (bladder) cell lines confirmed that JEG-3 expresses type II genes while T24 expresses exclusively type I. FBT11-II only recognizes JEG-expressed hCGβ. Placenta immunohistochemical studies confirmed that type II hCGβ expression is restricted to the syncytiotrophoblast. Immunoassays detected type II hCGβ in serum of patients with either nontrophoblastic cancers or fetal Down syndrome. ConclusionType II gene expression can be detected using FBT11-II. This specific recognition could improve the clinical usefulness of assays aimed at either managing aggressive tumors or screening for Down syndrome.

Evaluation of CGB, GNRH1, MET and KRT19 Genes Expression Profile as a Circulating Tumor Cells Marker in Blood of Cancer Patients

The aim of this study was to evaluate human chorionic gonadotropin beta subunit (CGB), gonadotropin releasing hormone type 1 (GNRH1), hepatocyte growth factor receptor (MET) and keratin 19 (KRT19) genes expression profile as a circulating tumor cells (CTC) marker in blood of cancer patients. Expression level of studied genes was assessed in peripheral blood of 122 patients with different types of cancers and 86 healthy volunteers using reverse transcription real time quantitative polymerase chain reaction (RT-qPCR). The result of the experiments demonstrated that in blood of cancer patients the level of MET transcripts showed positive correlation with KRT19 and negative correlation with GNRH1. In the control group negative correlation between CGB and GNRH1 was documented. What is more the level of CGB, MET and KRT19 expression was significantly higher in blood of cancer patients. Even though the analysis proved that studied genes are expressed in blood of both cancer patients and healthy volunteers, their expression level was highly heterogeneous. In order to interpret the results, the obtained data was log-transformed and fitted to multiplied normal distribution model using the maximal likelihood method. The results of this analysis showed elevated expression of CGB, MET and KRT19 together with extremely high levels of GNRH1 in blood of cancer patients which might indicate the presence of circulating tumor cells and increased risk of metastasis.

A Case of Ectopic Pregnancy—An Unusual Diagnostic Challenge and Lesson Learnt

Ectopic pregnancy has been increasing in frequency over the past 2 decades. The sudden rupture of ectopic pregnancy can lead to haemorrhagic shock and death if not diagnosed and treated in a timely fashion. The first step in the diagnosis of ectopic pregnancy is demonstration of pregnancy by means of a rapidly performed and sensitive qualitative urine test for the beta-subunit of human chorionic gonadotropin (beta-hCG). This case report demonstrates the importance of further consideration of the diagnosis of ectopic pregnancy in the setting of an unusual presentation like per rectal bleed and amenorrhea for three years in the presence of Mirena.

Laparoscopically diagnosed invasive mole presenting as acute hemoperitoneum.

A young multipara presented with acute abdominal pain. She had history of dilatation and evacuation for a missed miscarriage 2 months back. The diagnosis of ectopic pregnancy was made on the basis of clinical presentation and laboratory investigations. Laparoscopy was performed which revealed features of invasive mole. The procedure was converted to laparotomy and hysterectomy was performed. Patient recovered well. Histopathology confirmed the diagnosis of invasive mole. Follow-up till 12 weeks reported return to normal ßhCG (beta subunit of human Chorionic Gonadotropin) levels.

Review of ovarian tumors in children and adolescents: radiologic-pathologic correlation.

The incidence, histologic distribution, and clinical manifestations of ovarian tumors in the pediatric population are distinct from those in adults. Although ovarian neoplasms in childhood and adolescence are rare, the diagnosis should be considered in young girls with abdominal pain and a palpable mass. Differential diagnosis in children and adolescents with ovarian tumors should be conducted on the basis of unique clinical manifestations, elevated serum tumor marker levels, and distinctive imaging findings. Although the clinical manifestations are nonspecific and may overlap, they may assist in diagnosis of some types of ovarian tumors. Children who present with a palpable mass or symptoms of precocious puberty have a high likelihood of malignancy. Many ovarian tumors are associated with abnormal hormonal activity and/or abnormal sexual development. Elevated levels of serum tumor markers, including α-fetoprotein, the beta subunit of human chorionic gonadotropin, and CA-125, raise concern for ovarian malignancies. However, negative tumor markers do not exclude the possibility of malignancy. Identification of imaging features at ultrasonography, computed tomography, and magnetic resonance imaging can help differentiate benign from malignant ovarian tumors and, in turn, plays a crucial role in determining treatment options. At imaging, malignant ovarian tumors usually appear predominantly solid or heterogeneous and are larger than benign tumors. Because surgery is the primary treatment for ovarian tumors, ovarian salvage with fertility preservation and use of a minimally invasive surgical technique are important in children and adolescents.

Invasive or non-invasive prenatal genetic diagnosis?

About 40 years ago, invasive prenatal diagnosis techniques were introduced in obstetrics. Initially, amniocentesis was performed followed by placentacentesis, fetoscopy, fetal blood sampling (FBS), and chorionic villus sampling (CVS). These procedures, while invading the uterine environment, have made it possible to proceed with the retrieval of biological tissue of fetal origin for analysis and definitive diagnosis [6, 20, 21, 27]. The development of such techniques was facilitated by improvements in instrumentation and technology, and was further propelled by the advancement of cytogenetics and molecular genetic techniques [3, 13, 14, 22]. However, invasive prenatal diagnosis carries the inherent risk of fetal loss, which is low, but not negligible (amniocentesis and CVS approximately 0.3%–0.5% and FBS 1%–2%). In addition, there is a significant economic burden from the costs associated with laboratory techniques. Public health programs of nations interested in the utilization of invasive procedures have generally limited their use to high-risk cases, where the risk of procedure related loss is comparable to the risk of an affected fetus for a given condition [25, 26]. As a result, in the 1980s amniocentesis was offered to women at higher risk of trisomy 21 based on maternal age alone, if there was an increased risk due to prior complications or pre-existing conditions (i.e., chromosomal alterations in the parents or in prior offspring), or because of prenatal detection of fetal malformations or other abnormal ultrasound findings. The initial policies led to an offering of invasive prenatal diagnosis to 5% of all pregnant women (positive screen rate), with a 40% detection rate for trisomy 21. With the advent of biochemical screening tests using maternal serum in the second trimester, the “triple” and “quadruple screen”, the detection rate of trisomy 21 increased to 60% [28]. Meanwhile, with advancements in ultrasound, numerous reports were published that identified sonographic “markers” for trisomy 21, leading to additional screening with a “genetic ultrasound” in the second trimester [1]. However, second trimester ultrasound for the purposes of screening an unselected population never gained universal acceptance, and was primarily used in higher-risk populations (i.e., a woman with advanced maternal age that wished to avoid invasive testing). At this time, second trimester amniocentesis was the primary invasive diagnostic test practiced, while fetal blood sampling by cordocentesis was utilized when a diagnostic test was desired later in the second trimester, often in the setting of identification of a fetal anomaly in the second trimester ultrasound. As mean maternal age at childbirth continued to increase, especially in Western countries, alongside increasing scientific advancements, the number of indications for prenatal diagnosis rose. This trend led to an increased rate of invasive prenatal diagnosis, based on maternal age alone up to 15–20% in some nations, and spurred a search for new solutions. In the mid-1990s, an important turning point was the use of ultrasound to measure fetal nuchal translucency at 11–14 weeks [17], along with maternal serum biochemical screening, pregnancy associated plasma protein-A (PAPP-A), and free beta subunits of human chorionic gonadotropin (free beta-hCG), for the screening of trisomy 21 [24]. With a similar invasive testing rate as maternal age alone of 5%, the combined test led to a detection rate of approximately 90% for trisomy 21 (in the setting of a positive test, the odds of an affected fetus were 1:24). The introduction of the nuchal translucency measurement raised the issue of certification, reproducibility, and reliability of *Corresponding author: Giovanni Monni, Department of Obstetrics and Gynecology, Microcitemico Hospital, Prenatal and Preimplantation Genetic Diagnosis, Cagliari, via Edward Jenner SNC, Sardinia, Italy, Tel.: +39-706095546, Fax: +39-706095514, E-mail: prenatalmonni@tiscali.it Maria Angelica Zoppi, Ambra Iuculano, Alessandra Piras and Maurizio Arras: Department of Obstetrics and Gynecology, Microcitemico Hospital, Prenatal and Preimplantation Genetic Diagnosis, Cagliari, via Edward Jenner SNC, Sardinia, Italy

Germ cell and embryonal tumors

Germ cell tumors, which constitute approximately 3-5% of tumors of the central nervous system (CNS), can be subdivided into germinomas, embryonal carcinomas, yolk sac tumors, choriocarcinomas, teratomas and mixed germ cell tumors. The diagnosis of intracranial germ cell tumor is based on the clinical symptoms, detection of tumor markers, such as alpha fetoprotein (AFP) and the beta subunit of human chorionic gonadotropin (beta-hCG) in blood and cerebrospinal fluid (CSF), magnetic resonance imaging (MRI) of the brain and spinal cord, CSF cytology and histology. The diagnosis of a secreting germ cell tumor, i.e. a non-germinoma, can be made by the determination of AFP and hCG as tumor markers. Germinomas are radiosensitive but are equally as sensitive to chemotherapy. Teratomas of the CNS are mostly diagnosed in newborns and infants. The most decisive role in the treatment of teratomas is played by as complete a resection as possible. Chemotherapy and irradiation play a subordinate role.Embryonal tumors, which constitute approximately 15-20% of CNS tumors, include medulloblastomas, primitive neuroectodermal tumors (PNET) of the CNS and the atypical teratoid rhabdoid tumor of the CNS. Medulloblastoma is the most common malignant brain tumor in childhood and adolescence. The incidence peak is the fifth year of life with a male predisposition in a ratio of 1.5:1. Medulloblastomas constitute 12-25% of all pediatric CNS tumors and 30-40% of pediatric tumors of the posterior cranial fossa. At the time of diagnosis evidence of dissemination in the CSF cavity is found in approximately 40% of patients. The extreme cell density makes medulloblastomas hyperdense in computed tomography (CT) and can therefore be differentiated from hypodense astrocytomas. The PNETs are histologically related to medulloblastomas, pineoblastomas, atypical teratoid rhabdoid tumors and peripheral neuroblastomas. They are relatively rare in children constituting less than 5% of supratentorial neoplasms. Patients are mostly clinically conspicuous due to macrocephalus and signs of brain pressure and/or seizures. In native CT the solid components of PNETs show a hyperdensity compared to the surrounding brain parenchyma probably due to the high cell density. Cysts and calcification are often detectable. The survival rate of children with CNS tumors has continuously increased in recent years. When corresponding clinical symptoms appear, such as headache, nausea or vomiting when fasting, all of which are evidence of increased intracranial pressure, MRI should be carried out as quickly as possible. Children should be treated in centers with departments of pediatric oncology and hematology and within the framework of studies.

Methylation status of human chorionic gonadotropin beta subunit promoter and TFAP2A expression as factors regulating CGB gene expression in placenta

To evaluate mechanisms regulating the expression of CGB genes in placental tissues from uncomplicated pregnancies and chorionic samples from spontaneous miscarriages. Molecular analyses in human samples. Laboratory of molecular biology. Nine placental samples from term deliveries and 21 chorionic samples from miscarriages at 7-13weeks of gestation. None. The expression level of CGBs and genes encoding SP1, SP3, and AP2 transcription factors was analyzed using quantitative polymerase chain reaction (qPCR). The methylation status of the CGBs' promoter regions was determined using methylation-specific PCR. The experiments showed significant differences in CGBs' expression and their regulation between placental and chorionic tissues. In placental tissues and chorionic tissues from 7 to 9weeks of gestation, the expression level of CGBs was shown to be associated with the amount of TFAP2A transcripts. It was also demonstrated that variation in the expression level of CGB genes relies on changes in methylation of CGB3-9 and CGB1-2 promoter sequences. During pregnancy, regulation of hCG beta subunit genes expression correlates with both methylation of their promoters and TFAP2A expression level. The results suggest that these factors may be very influential in the early stages of pregnancy and may be associated with pregnancy outcome.