hCG Production Research Articles

Individual deviations in human chorionic gonadotropin concentrations during pregnancy

Serum and urine human chorionic gonadotropin (hCG) vary greatly during the course of pregnancy. We investigated the cause of this variation. Eighty-two women provided daily urine samples during the first 6 weeks of gestation. First-void urine samples were monitored for luteinizing hormone (LH) and hCG. Variation was wide when pregnancy hCG anchoring to the last menstrual period (variation 677 ± 786-fold) or to LH peak (variation 810 ± 936-fold). When pregnancy was anchored to the day of implantation (variation 187 ± 123-fold) variation was significantly reduced (P < .00005). Individual differences in the rate of hCG production were examined. hCG production ranged from 1.52-fold to 2.92-fold per day. Rate differences in hCG were also a major source of hCG variation. Two factors are responsible for the wide fluctuation in hCG concentrations, first dating pregnancies to the start of the last menstrual period rather than the timing of implantation and second, individual pregnancy differences in the rate of hCG production.

Characterisation of transient benign hCG elevations in women following chemotherapy for GTT

The occurrence of post-chemotherapy transient low level hCG elevations has been observed in a number of women treated for gestational trophoblastic tumours (GTT). The authors reviewed the records of patients treated at Charing Cross Hospital over the last 10 years and identified those with a benign rise in hCG. Stored serum samples were assayed for hCG, LH, FSH and oestradiol at varying points during patient management. The endocrine profile in patients experiencing benign hCG elevation is comparable with that seen in post-menopausal women, with low oestradiol, combined with greatly elevated LH and FSH levels. In contrast, women with genuine disease relapse as the cause of their post-chemotherapy hCG elevation had normal or only minor elevations of LH and FSH. These findings support the observation that a major rise in LH and FSH can be used as an indicator for benign pituitary hCG production in patients experiencing a low level rise in hCG levels following chemotherapy for GTT.

Effect of St. John's Wort Standardized Extract and Hypericin on In Vitro Placental Calcium Transport

Hypericum perforatum L., known as St. John's wort (SJW), is widely used in human therapeutics for wound healing and to treat depression; however, its recommendation during pregnancy is controversial. Hypericin, a polycyclic quinone isolated from this plant, has been studied and used to standardize the plant extracts. The objective of the present study was to examine the effect of SJW and hypericin on in vitro placental Ca(2+) transport. Cell viability, human chorionic gonadotrophin (hCG) production, Ca(2+) uptake, and Ca(2+) transport proteins expression analysis were conducted using the JEG-3 cell line. Toxicity of SJW was seen at high concentrations (>or=150 microg/mL), but no effect on hCG production was observed using SJW (25 microg/mL) or hypericin (7.5 and 75 ng/mL). The results showed that cells treated with both SJW and hypericin exhibited increased intracellular Ca(2+) concentration after long-term (24-hour) but not short-term (10-minute) period incubation. A significant decrease in translationally controlled tumor protein Ca(2+) handling protein was seen only with SJW-treated cells. Hypericin increased the protein expression of the transient receptor potential vanilloid 6 Ca(2+) channel and 28-kDa calcium-binding protein and decreased that of plasma membrane Ca(2+)-ATPase 1/4. In conclusion, SJW and hypericin can increase the trophoblast internal Ca(2+) concentration through regulating the protein expression of the Ca(2+) transport system, and their intake during pregnancy is still a point of concern.

Possible role of the exchange protein directly activated by cyclic AMP (Epac) in the cyclic AMP-dependent functional differentiation and syncytialization of human placental BeWo cells

The mononuclear villous cytotrophoblast (CTB) differentiates and fuses to the multinucleated syncytiotrophoblast (STB), which produces hCG and progesterone. cAMP-mediated intracellular pathways are involved in the process of endocrine differentiation and fusion (syncytialization). The exchange protein directly activated by cAMP (Epac) is a mediator of cAMP signaling. We examined the differential roles of Epac and protein kinase A (PKA) signaling in the cell fusion and differentiation of trophoblast-derived BeWo cells. Epac1 and Epac2 were localized in human placental tissue (n = 9) by immunohistochemistry. The PKA-selective cAMP analog (N(6)-phenyl-cAMP, Phe) or Epac-selective cAMP analog (CPT) was tested for effects on hCG and progesterone production, and syncytialization in BeWo cells. The effect of knockdown of Epac or its downstream target molecule (Rap1) on syncytialization was evaluated. Epac1 and Epac2 proteins were expressed in villous CTB, STB, stroma, blood vessels and extravillous CTB of the placenta. Phe increased the expression of hCG alpha/beta mRNA and secretion of hCG protein in BeWo cells (P < 0.01 versus control). CPT-stimulated production of hCG (P < 0.05), albeit to a lesser extent than Phe. Progesterone production was also enhanced by Phe or CPT (P < 0.01 and P < 0.05, respectively). CPT or a stable cAMP analog (dibutyryl-cAMP: Db) increased the number of syncytialized BeWo cells (P < 0.01), whereas Phe did not stimulate fusion. CPT- or Db-induced syncytialization was observed, even in the presence of a PKA inhibitor. Knockdown of Epac1 or Rap1 repressed the Db-, CPT- or forskolin-induced cell fusion. The Epac signaling pathway may be associated with the cAMP-mediated functional differentiation and syncytialization of human trophoblasts.

Pituitary hCG production and cerebral tuberculosis mimicking disease progression during chemotherapy for an advanced ovarian germ cell tumour

BackgroundOvarian germ cell tumours (OGCT) are rare but are usually curable with chemotherapy, even when presenting with advanced disease. The majority of OGCT produce the tumour markers, hCG and/or AFP which can be helpful in the diagnosis and monitoring the response to treatment.Case PresentationIn this case of a 36 year old woman, the elevated hCG level at presentation was helpful in making a clinical diagnosis of OGCT in a patient too unwell to permit a tissue diagnosis.Cisplatin based combination chemotherapy produced an initial normalisation of the hCG level, but later in treatment the patient developed new cerebral lesions and a rising serum hCG suggestive of disease progression.Further investigations suggested that the CNS lesions were cerebral TB and that the low levels of hCG elevations was likely to be pituitary in origin. Chemotherapy treatment was continued along with anti-tuberculous therapy and 24 months after successful completion of therapy the patient remains disease free.ConclusionsIn the treatment of cancer patients it may be helpful to consider the potential non-malignant causes of new CNS lesions and that low hCG elevations may result from physiology rather than pathology in selected cases.

Role of GnRH–GnRH receptor signaling at the maternal-fetal interface

To investigate the expression and function of GnRH and GnRH receptor (GnRHR) subtypes at the maternal-fetal interface. In vitro experiments using freshly isolated human trophoblast cells, decidual stromal cells (DSCs), and immortalized cell lines. University teaching hospital. Placenta-fetal membranes from term deliveries. Human trophoblast and DSCs were isolated, purified, and cultured. Expression of GnRH-I, GnRH-II, and GnRHR-I mRNA and protein in human trophoblast cell lines and tissues were evaluated by reverse-transcription polymerase chain reaction and Western blot. The effect of GnRH-I and -II on the production of select cytokines (hCG, interleukin [IL] 8, IL-6, matrix metalloproteinase 3, monocyte chemoattractant protein 1, vascular endothelial growth factor, soluble Fms-like tyrosine kinase 1, urokinase-type plasminogen activator, and plasminogen activator inhibitor 1) were measured by ELISA and normalized for protein content. GnRH-I, GnRH-II, and GnRHR-I mRNA and protein were identified in trophoblasts and decidua. GnRH-I and -II stimulated hCG production by trophoblast and trophoblast-derived cell lines in a dose-dependent fashion (e.g., 2.8-fold, from 2.5 ± 0.5 to 7.0 ± 0.4 ng/mg protein per 24 h, for 1,000 nmol/L GnRH-I and 2.4-fold, from 2.5 ± 0.5 to 6.1 ± 0.6 ng/mg protein per 24 h, for 1,000 nmol/L GnRH-II) without affecting the production of other cytokines. Trophoblasts and decidua express GnRH-I, GnRH-II, and GnRHR-I mRNA and protein. GnRH-I and -II selectively stimulate hCG production by trophoblast cells without altering the production of select cytokines by trophoblasts or decidua. The role of GnRH-GnRHR signaling at the maternal-fetal interface therefore appears to be limited to the regulation of trophoblast hCG production.

Normal Production of Human Chorionic Gonadotropin in Perimenopausal and Menopausal Women and After Oophorectomy

The normal pituitary production of human chorionic gonadotropin (hCG)alongside luteinizing hormone, and its presence in women after bilateral oophorectomy, during perimenopause and menopause, as measured in serum and urine, has been known for 30 years and is described in numerous publications. Last year our group discussed this finding in a correspondence to the editor in the March 15th issue of New England Journal of Medicine, yet the misinterpretation of low-level hCG in these women seems to have increased in magnitude. This is an outcomes study of 36 cases of menopausal hCG referred to the USA hCG Reference Service over a 1-year period, from March 2007 to March 2008. Eight cases occurred in women after oophorectomy, 28 were women in menopause/perimenopause. Surgery was postponed in 5 (14%) of 36 cases, and in 3 cases (8%), chemotherapy was unnecessarily administered. In 2 cases, computed tomography scans were cancelled. The average hCG detected was 10 +/- 7.2 IU/L in cases receiving an oophorectomy and 9.8 +/- 6.7 in perimenopause and 11 +/- 6.2 IU/L in menopause cases. Low-level hCG production in woman in physiologic perimenopause, in menopause, or in women with prior bilateral oophorectomy is a normal biologic and biochemical phenomenon. Management protocols in all fields need to be changed to accept pituitary hCG as normal and recognize the clinical maneuvers that will secure the diagnosis. Understanding this physiology will avoid needless delays in necessary therapies such as organ transplant procedures and will limit the misadventure of prescribing unnecessary cancer treatments.

Expression and Regulation by PPARγ of hCG α- and β-subunits: Comparison between Villous and Invasive Extravillous Trophoblastic Cells

During human placental development trophoblast follows two differentiation pathways: the extravillous (EVCT) and the villous cytotrophoblasts (VCT) that display different phenotypes and functions. It is well established that human chorionic gonadotropin hormone (hCG) is mainly secreted by the endocrine VCT (syncytiotrophoblast) into the maternal compartment and stimulates the formation of the syncytiotrophoblast (ST) in an autocrine manner. We recently reported that the invasive EVCT also produces hCG that promotes trophoblast invasion in vitro. Herein, we compared hCG gene expression in primary culture of villous and extravillous trophoblasts obtained from the same first trimester human chorionic villi and differentiated in vitro into ST and invasive EVCT, respectively. Total hCG, free α and free β subunits were quantified in cell supernatants by immunometric assays and normalized to DNA content. α and β transcript levels were quantified by Q-PCR and normalized to cytokeratin 7. We show that free α-, free β-subunits and total hCG are differently expressed and secreted by the two trophoblast subtypes during their differentiation in vitro. We found an α/β ratio 100 times lower in invasive EVCT in comparison to the ST suggesting that β subunit may not be step limiting for hCG production in EVCT. Finally we investigated the regulation of hCG gene expression by PPARγ, a nuclear receptor that controls trophoblast differentiation and invasion. Interestingly, activation of PPARγ by the agonist rosiglitazone gave opposite results in the endocrine VCT and invasive EVCT: α and β subunit transcript levels and protein secretions were up regulated in VCT, whereas they were down regulated in EVCT. Our results demonstrated that hCG gene expression is differentially regulated in the two trophoblast lineages during their in vitro differentiation and modulated in an opposite way by PPARγ.

A Placental Protective Role for Trophoblast-Derived TNF-Related Apoptosis-Inducing Ligand (TRAIL)

Recent studies show that apoptosis, programmed cell death, plays an important role in the normal development of the human placenta and that an altered balance between proliferation and apoptosis of villous trophoblasts is associated with abnormal pregnancies. The TNF-related apoptosis-inducing ligand (TRAIL) is a molecule belonging to TNF superfamily. The role of TRAIL and its Death Receptor 5 (DR5) in regulating villous trophoblast cell turnover in normal and pathologic pregnancies remains to be explored. In order to elucidate the role of TRAIL in the regulation of placental growth, primary cytotrophoblast cells were isolated from normal term placentas ( n = 13) and cultured for 18 and 66 h to generate mononucleate and multinucleate trophoblasts, respectively. The protein expression and localisation of TRAIL and DR5 were determined by Western blotting and immunofluorescence. Secreted sTRAIL was also measured by ELISA. Trophoblast apoptosis was measured by TUNEL in the presence of recombinant TRAIL (rTRAIL), and DR5 relocalisation was assessed by immunostaining after 18 h exposure to TNFα. We demonstrated that TRAIL protein expression and the secretion of soluble TRAIL (sTRAIL) were down-regulated in syncytialised villous trophoblasts and that sTRAIL was independent of biochemical differentiation, as TRAIL-neutralizing antibody (2E5) failed to influence hCG production. TRAIL immunoreactivity was detected in mono- and multinucleated trophoblast cells and localised to the cytoplasm and cellular membranes – more intense staining was associated with apoptotic nuclei. rTRAIL failed to induce apoptosis in trophoblasts cells owing to the nuclear localisation of DR5. However, TNFα treatment caused the redistribution of intracellular DR5 to the cell surface, potentiating apoptotic susceptibly to exogenously administered rTRAIL. These findings highlight a mechanism by which TRAIL and DR5 serve to protective trophoblasts in normal development, but may be activated in conditions of excessive TNFα.

The primate placenta and human chorionic gonadotropin.

In the primate placenta various peptide and proteohormones are synthesized which control growth and development of the fetus as well as the exchange of nutrients and metabolic products between the mother and the fetus. In humans, maintenance of pregnancy in the first trimester depends on the synthesis of the bioactive glycoprotein hormone human chorionic gonadotropin (hCG). It is expressed in placenta by the syncytiothrophoblast of early pregnancy. In cell culture, hCG production seems to mark a certain step in the process of differentiation of cytotrophoblasts and choriocarcinoma cells. It is neither understood how hCG synthesis is initiated and maintained at the beginning of gestation nor what control mechanisms are responsible for the down-regulation of the synthesis at the end of the first trimester. Besides a long list of various other substances which have been described to act as intrinsic placental stimulators of hCG biosynthesis, gonadoliberin and gamma-aminobutyric acid seem to play an important role. This establishes to some extent an analogy to the regulation of gonadotropin synthesis in the central nervous system. Recently, a full-length form of functional LH/hCG receptors of approximately 80 kD has been found in term placenta suggesting autoregulation as a regulatory principle of hCG biosynthesis. In the first trimester placenta as well as in choriocarcinoma cells a truncated form (50 kd) of LH/hCG receptors seems to exist. In these cases, exogenous hCG was unable to down-regulate its own synthesis. The carbohydrate moiety of hCG influences folding, subunit assembly, circulatory half-life, receptor interaction and biological response. A surplus of glycosylation may prevent subunit assembly. Experimental deglycosylation induces a different conformation of hCG, which partly acquires antagonistic properties. Recent results indicate that cAMP, which increases transcription and mRNA stability, also expands the N-glycosylation capacity and thus may accomplish an over-all coordination of hCG biosynthesis including post-translational events.

New discoveries on the biology and detection of human chorionic gonadotropin.

Human chorionic gonadotropin (hCG) is a glycoprotein hormone comprising 2 subunits, alpha and beta joined non covalently. While similar in structure to luteinizing hormone (LH), hCG exists in multiple hormonal and non-endocrine agents, rather than as a single molecule like LH and the other glycoprotein hormones. These are regular hCG, hyperglycosylated hCG and the free beta-subunit of hyperglycosylated hCG.For 88 years regular hCG has been known as a promoter of corpus luteal progesterone production, even though this function only explains 3 weeks of a full gestations production of regular hCG. Research in recent years has explained the full gestational production by demonstration of critical functions in trophoblast differentiation and in fetal nutrition through myometrial spiral artery angiogenesis.While regular hCG is made by fused villous syncytiotrophoblast cells, extravillous invasive cytotrophoblast cells make the variant hyperglycosylated hCG. This variant is an autocrine factor, acting on extravillous invasive cytotrophoblast cells to initiate and control invasion as occurs at implantation of pregnancy and the establishment of hemochorial placentation, and malignancy as occurs in invasive hydatidiform mole and choriocarcinoma. Hyperglycosylated hCG inhibits apoptosis in extravillous invasive cytotrophoblast cells promoting cell invasion, growth and malignancy. Other non-trophoblastic malignancies retro-differentiate and produce a hyperglycosylated free beta-subunit of hCG (hCG free beta). This has been shown to be an autocrine factor antagonizing apoptosis furthering cancer cell growth and malignancy.New applications have been demonstrated for total hCG measurements and detection of the 3 hCG variants in pregnancy detection, monitoring pregnancy outcome, determining risk for Down syndrome fetus, predicting preeclampsia, detecting pituitary hCG, detecting and managing gestational trophoblastic diseases, diagnosing quiescent gestational trophoblastic disease, diagnosing placental site trophoblastic tumor, managing testicular germ cell malignancies, and monitoring other human malignancies. There are very few molecules with such wide and varying functions as regular hCG and its variants, and very few tests with such a wide spectrum of clinical applications as total hCG.

Epithelial Human Chorionic Gonadotropin Is Expressed and Produced in Human Secretory Endometrium During the Normal Menstrual Cycle1

The objective of this study was to determine whether beta human chorionic gonadotropin (hCG) (CGB) subunits and alpha hCG (CGA) subunits are expressed and the hCG dimer is produced in normal human cyclic endometrium. Endometrial specimens were collected for histological dating from women undergoing treatment in our division of human reproduction. RNA from normal secretory endometrium was extracted, and CGB and CGA gene expression was assessed by semiquantitative PCR. Adequate secretory endometrial specimens were homogenized using protease inhibitors. Proteins present in the supernatant were separated electrophoretically, and molecular hCG isoforms were detected by Western blot. The supernatant hCG concentrations were measured by ELISA. We characterized hCG and leukocytes in endometrial specimens by immunohistochemistry. Uterine flushing was performed to confirm endometrial hCG secretion into the uterine fluid. A full-length CGB mRNA encompassing the exon 1 promoter region and the structure exons 2 and 3 (including the C-terminal peptide) was expressed in normal secretory endometrial specimens (similar to CGA) during the early secretory phase of the menstrual cycle, up to an optimum at the midsecretory to late secretory phases. In homogenate supernatants obtained from normal secretory endometrium, hormone concentrations of dimeric hCG were approximately 5 mU per 10 mg of tissue, compared with considerably smaller concentrations of corresponding single free CGB subunit. Single chains of CGB, CGA, and dimeric molecular hCG isoforms were found in endometrial specimens by Western blot. Glandular endometrial hCG production is demonstrated immunohistochemically, with an increase toward the late secretory phase vs. the early secretory phase of the normal secretory menstrual cycle. However, glandular hCG release is diminished or absent in the dyssynchronous or missing endometrial secretory transformation. Endogenous endometrial hCG may be important for implantation and maintenance of pregnancy.

Not even a little bit pregnant – ca colon as cause of false positive pregnancy test

<h3>Introduction</h3> Pregnancy testing is usually performed in women of reproductive age prior to any procedure which could potentially harm a developing embryo. Occasionally, false positive pregnancy tests may occur and have the potential to cause not only delay in treatment but also great consternation for the patient concerned. <h3>Patient</h3> A 38-year-old female with advanced colorectal carcinoma (Duke's stage 4) had serum beta human chorionic gonadotropin (hCG) tested prior to receiving chemotherapy. hCG was 74 IU/L which was interpreted as unambiguously positive. Chemotherapy was therefore delayed in spite of the patient believing she could not be pregnant. <h3>Investigations</h3> The luteinising hormone (LH) and follicle stimulating hormone (FSH) were normal for a woman of reproductive age. The initial result was confirmed on two alternate analysers and repeated 1 week later when it had risen to 84. Elimination of heterophile antibodies by use of Scantibodies and urine hCG confirmed the presence of free hCG. Performance of immunohistochemical staining of the original tumour collected 31 months earlier for hCG revealed patchy production of hCG in about 5% of the tumour population. This was seen predominantly at the advancing margin of the tumour. <h3>Discussion</h3> An hCG of 74 IU/L is normally considered unambiguously positive and the absence of heterophile antibodies or peri/post-menopausal source of pituitary hCG appeared to further increase the possibility of a normal or ectopic pregnancy. However the failure of the hCG to rise rapidly, a normal abdominal ultrasound and the protestations of the patient made further investigation necessary. The demonstration of hCG producing tumour tissue confirmed the likely origin of the circulating hCG. <h3>Conclusion</h3> Laboratories must be constantly aware of the common sources of false positive pregnancy tests including heterophile antibodies, ‘macro' hCG, pituitary hCG in peri/post menopausal women and tumour tissue itself. The production of hCG by germ cell tumours and teratomas is well recognised but the production by gastrointestinal cancers is much less well recognised. Recent reports suggest that non-trophoblastic tumours producing hCG are associated with a poor prognosis but a technique to stimulate anti-hCG antibodies may provide therapeutic promise.

Grossesse et lupus systémique

Pregnancy is widely authorized in systemic lupus erythematosus (SLE). Fertility is similar in SLE and in the general population although the age of menarche seems higher. Some cases of sterility might be attributed to SLE because of autoimmune ovaritis or antiphospholipid antibodies (aPL). These antibodies might lead to endothelial activation and thrombosis by influencing homeostasis, complement activation, inhibition of protein C and annexin V. They might have a deleterious effect on embryonic implantation by adhesion to the trophoblast, inhibition of invasion and placentation and decreased hCG production. The most important part of sterility seems secondary to the use of cyclophosphamide and might be prevented by acetate leuprolide administration. Maternal morbidity seems correlated to SLE activity (controlled by pregnancy planning), hypertension, preeclampsia, Hemolysis, Elevated Liver Enzymes, Low Platelets (HELLP) syndrome, therapy and aPL. Hydroxychloroquine (HCQ) should be maintained throughout pregnancy. Aspirin is prescribed alone in patients with asymptomatic aPL and in addition to heparin if there is a history of thrombosis or fetal loss with aspirin. Fetal and neonatal morbidity correlate with prematurity, adverse effects or maternal steroid therapy and maternal anti-SSA antibodies with 1 to 2% risk of congenital atrioventricular block. Abnormal obstetrical echography-doppler examination is the best predictor of pregnancy outcome. Abnormal ombilical artery flow on the second trimester echodoppler examination and history of thrombophlebitis predict fetal or neonatal death. Abnormal uterine notch on the second trimester echodoppler examination predicts adverse pregnancy outcome. Except for the preventive therapy of congenital atrioventricular block, modalities of SLE pregnancy monitoring and therapy are now well established.

Comparison of cell cycle regulatory gene mRNA in three different types of human trophoblasts and effect of transforming growth factor

Identifying the factors responsible for reducing the proliferation, syncytialization, and invasiveness of trophoblast tissues, as seen with preeclampsia, intrauterine growth restriction, and spontaneous miscarriage, is a current challenge in reproductive biology. These factors, transforming growth factor (TGF)-beta as an example, can work by altering trophoblast differentiation or proliferation. We therefore investigated and compared specific markers of trophoblast proliferation and differentiation in three commonly used trophoblast tissue cell models, and also investigated the influence of TGF-beta on these markers. In this study, we isolated human trophoblasts from first trimester and term placentas, and additionally used human choriocarcinoma cells (JEG-3). Baseline values of human chorionic gonadotropin (hCG) secretion and relative mRNA levels of cell cycle regulators (cyclin E, p21, p27, and p57) were investigated for each cell type. We also investigated the influence of TGF-beta on these parameters. Quantitative and longitudinal production of hCG differed between the three cell types. Significantly different amounts of cyclin E, p21, p27, and p57 mRNA were demonstrated within each cell type, as well as between all the cell types, throughout the culture time period. Each trophoblast type demonstrated a reduction of hCG secretion in response to TGF-beta. TGF-beta did not show a consistent effect on the cell cycle mRNA of any of the cell types. We were able to characterize and compare the differential production of hCG, as well as the differential expression of cell cycle-associated mRNA of early trophoblasts, term trophoblasts, and choriocarcinoma cells. The production of hCG was altered by TGF-beta, although mRNA levels were not markedly altered by TGF-beta.

Stimuli through Toll‐like receptor (TLR) 3 and 9 affect human chorionic gonadotropin (hCG) production in a choriocarcinoma cell line

Toll-like receptors (TLR) are expressed mostly on mucosal innate immune cells which recognize microorganism-associated structures, the so called pathogen-associated molecular patterns (PAMP). Recently, possible roles of TLR in placental tissue, especially their role in protecting the conceptus from maternal infection, have been discussed, but the physiopathological roles of TLR in placental functions have yet to be examined. The choriocarcinoma cell line BeWo was used as a model of trophoblast stem cells. First, the expression of TLR 1-9 on BeWo cells was investigated using reverse transcriptase-polymerase chain reaction. Then, the cells were treated with TLR 1-9 agonists, using the human TLR 1-9 agonist kit (Pam3CKS4, HKLM, Poly (I:C), Escherichia coli K12 lipopolysaccharide (LPS), Salmonella typhimurium flagellin, FSL1, imiquimod, ssRNA 40, and ODN 2006). The concentration of human chorionic gonadotropin (hCG) in the cell culture supernatants was examined on ELISA. BeWo cells constitutively expressed TLR 1-9 mRNA in the presence or absence of forskolin. The TLR 3 agonist, Poly(I:C), and the TLR 9 agonist, CpG-DNA (ODN2006), upregulated hCG production but had minimal effects in the absence of forskolin. Stimulation with the other TLR agonists produced no remarkable increase, whether in the presence or absence of forskolin. TLR 3 and TLR 9 agonists upregulated hCG production in BeWo cells. The present observations suggest alternative roles of TLR signaling in the control of placental functions and blastocyst-endometrial dialogue, as well as in pathophysiology, including for infertility and ectopic pregnancy.

Dedifferentiated Liposarcoma of the Retroperitoneum with Extensive Leiomyosarcomatous Differentiation and -Human Chorionic Gonadotropin Production

Dedifferentiated liposarcomas may display a variety of “heterologous” lines of differentiation, including osseous, vascular, skeletal, and/or smooth muscular. There have been six previously reported examples of leiomyosarcomas associated with high levels of serum human chorionic gonadotropin (hCG) production, comprised of cases originating from the retroperitoneum, spermatic cord, small intestine, and uterus. This report describes the first example of a dedifferentiated liposarcoma that combined both of the aforementioned features: extensive heterologous (leiomyosarcomatous) differentiation and β-hCG production (maximum serum levels 1046 mIU/ml, reference <5 mIU/ml). The tumor, which originated in the retroperitoneum in the region of the right kidney, was rapidly progressive and ultimately fatal within three months of its diagnosis. In addition to characteristic morphologic features, lipogenic and smooth muscle differentiation were confirmed with immunohistochemical stains for MDM2 and smooth muscle actin, respectively. The tumor also displayed diffuse immunoreactivity for β-hCG in both primary and metastatic sites. This case further expands the clinicopathologic spectrum of lipogenic tumors.

Urinary hCG patterns during the week following implantation

Human chorionic gonadotrophin (hCG) is used to monitor pregnancy status. Yet the pattern of hCG excretion in the first week following implantation has not been adequately described. Therefore the aim of this study was to describe the average profile of hCG and its variability during the 7 days following estimated implantation in a population of naturally conceived pregnancies. We measured daily hCG concentrations in first-morning urine for 142 clinical pregnancies from women with no known fertility problems. Mixed-effects regression models were used to estimate the hCG trajectory and its variability in relation to pregnancy outcomes. hCG rose 3-fold between the day of detection and the next day (95% CI = 2.7-3.4). The relative rate of rise decreased thereafter, reaching 1.6-fold (95% CI = 1.5-1.8) between days 6 and 7. HCG levels followed a log-quadratic trajectory, and the patterns of rise were unrelated to number of fetuses, risk of spontaneous abortion or sex of the baby. Later implantations (after 10 luteal days) produced slower rates of increase. Although mean hCG follows a log-quadratic trajectory during the first week of detectability, there is high variability across pregnancies. Later implantation may reflect characteristics of the uterus or conceptus that slow hCG production.

401: Thrombin suppresses hCG production by human syncytiotrophoblast: A mechanism by which first trimester bleeding causes abortion

Bleeding in the first trimester is associated with spontaneous pregnancy loss, but whether this bleeding causes the abortion or is simply a sign of an already unsuccessful implantation is not clear. Adequate production of hCG by syncytiotrophoblast is critical to the maintenance of early pregnancy by sustaining progesterone production by the corpus luteum. We propose that bleeding may cause abortion by interfering with hCG-progesterone signalling. This study investigates the effect of thrombin on hCG production by syncytiotrophoblast in vitro.

409: Effect of thrombin on GnRH-I and GnRH receptor (GnRHR) gene expression in human cytotrophoblast cells

GnRH is critical in regulating mammalian reproduction. In addition to its action in the pituitary, GnRH acts through its receptor (GnRHR) on the cell surface of extra-pituitary tissues, including placenta and decidua, where it exerts local autocrine/paracrine functions. The function of GnRH signaling at the maternal-fetal interface is not well understood, but it has been implicated in decidualization, trophoblast invasion and angiogenesis, and regulation of hCG production. Placental abruption is associated with adverse pregnancy outcome, but the mechanism is poorly defined.