Assessment of therapeutic effect of human choriogonadotropin in a chemical cystitis model.

In this study, female rats induced with chemical cystitis were administered the hormone human choriogonadotropin (HCG), and it was aimed to reveal the usefulness of HCG in the treatment of interstitial cystitis/bladder pain syndrome. The materials for this study were 32 Wistar albino female rats. The study groups were formed as follows: the cystitis group (Group 1), the cystitis + HCG protection group (Group 2), the cystitis + HCG treatment group (Group 3), and the control group (Group 4), with eight rats in each group. In this study, blood and urine samples were taken from the rats, they were euthanized, and their bladders were removed for glutathione, malondialdehyde, tumor necrosis factor alpha, and interferon gamma measurements. It was observed that tissue damage in Group 2 was lower than that in the other two groups. Glutathione levels in Groups 2 and 4 were significantly higher than in Groups 1 and 3 (p = 0.01). Malondialdehyde levels of Groups 2 and 4 were significantly lower than the values in Groups 1 and 3 (p < 0.001). When the cystitis groups were compared in terms of their interferon gamma and tumor necrosis factor alpha levels, the lowest interferon gamma and tumor necrosis factor alpha levels were detected in Group 3. It was found that HCG has positive effects on experimental cystitis in rats. This study revealed that HCG should be researched as a therapeutic agent and formed a step for studies to be carried out on this subject.

A 3D co-culture microtissue model of the human placenta for nanotoxicity assessment.

There is increasing evidence that certain nanoparticles (NPs) can overcome the placental barrier, raising concerns on potential adverse effects on the growing fetus. But even in the absence of placental transfer, NPs may pose a risk to proper fetal development if they interfere with the viability and functionality of the placental tissue. The effects of NPs on the human placenta are not well studied or understood, and predictive in vitro placenta models to achieve mechanistic insights on NP-placenta interactions are essentially lacking. Using the scaffold-free hanging drop technology, we developed a well-organized and highly reproducible 3D co-culture microtissue (MT) model consisting of a core of placental fibroblasts surrounded by a trophoblast cell layer, which resembles the structure of the in vivo placental tissue. We could show that secretion levels of human chorionic gonadotropin (hCG) were significantly higher in 3D than in 2D cell cultures, which indicates an enhanced differentiation of trophoblasts grown on 3D MTs. NP toxicity assessment revealed that cadmium telluride (CdTe) and copper oxide (CuO) NPs but not titanium dioxide (TiO2) NPs decreased MT viability and reduced the release of hCG. NP acute toxicity was significantly reduced in 3D co-culture MTs compared to 2D monocultures. Taken together, 3D placental MTs provide a new and promising model for the fast generation of tissue-relevant acute NP toxicity data, which are indispensable for the safe development of NPs for industrial, commercial and medical applications.

Toxicity assessment on trophoblast cells for some environment polluting chemicals and 17ß-estradiol.

The identification of reproductive toxicants is a major scientific challenge for human health. We investigated the effects of a selected group of environmental polluting chemicals mostly provided with estrogenic activity on the human trophoblast cell lines BeWo and HTR-8/SVneo. Cells were exposed for 24h to various concentrations (from 0.1 pM to 1 mM) of atrazine (ATR), diethylstilbestrol (DES), para-nonylphenol (p-NP), resveratrol (RES) and 17 ß-estradiol (E2) and assayed for cell viability and human beta-Chorionic Gonadotropin (ß-hCG) secretion. Decrease of cell viability as respect to control, vehicle-treated, cultures was obtained for all chemicals in the concentration range of 1 µM-1 mM in both cell types. A parallel decrease of ß-hCG secretion was observed in BeWo cells, at 1 µM-1 mM concentrations, with the only exception of ATR which caused an increase at concentrations up to 1mM. ß-hCG release was also unexpectedly inhibited by ATR, DES, p-NP and RES at non-toxic (pM-nM) concentrations. These findings raise concern about the negative, potential effects of various environmental polluting chemicals on pregnancy success and fetal health.

Toward a fast, easy, and versatile immobilization of biomolecules into carbon nanotube/polysulfone-based biosensors for the detection of hCG hormone.

The aim of this study was the fabrication and characterization of biomembranes by the phase inversion (PI) method followed by their subsequent casting onto screen-printed electrodes (SPE) for biomedical applications. The combination of multiwalled carbon nanotubes (MWCNT) as a transducer with polysulfone (PSf) polymer enables easy incorporation of biological moieties (hormones or antibodies), providing a 3D composite with high electrochemical response to corresponding analytes. Antibody/MWCNT/PSf biosensors were characterized by confocal scanning laser microscopy (CSLM), scanning electron microscopy (SEM), and electrochemical methods. For biomedical purposes, human chorionic gonadotropin (hCG) hormone was tested by competitive immunoassay. The detection limit was determined to be 14.6 mIU/mL with a linear range up to 600 mIU/mL. We concluded that the easy and fast incorporation of biomolecules by the PI method, as well as their stability and distribution throughout the 3D polysulfone composite, are testament to the utility for the versatile fabrication of biosensors for clinical diagnosis.

[Assessment of expression of luteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptor (LH/hCGR) and hCG protein in ovarian cancer tissues]. / Ocena ekspresji receptora hormonu luteinizujacego (LH)/ludzkiej gonadotropiny kosmówkowej (hCG) (LH/hCGR) oraz bialka hCG w raku jajnika.

OBJECTIVES: Ovarian cancers still remain a significant worldwide epidemiological problem. The vast majority of newly diagnosed cases are in advanced clinical stages, and the five-year survival rate in all clinical stages amounts up to less than fifty percent. A better understanding of the biology of ovarian cancer can bring us closer to successful treatment and recovery. The aim of this study was to evaluate the expression of luteinizing hormone (LH)/human chorionic gonadotropin (hCG) receptor (LH/hCGR) and the expression of hCG protein in the ovarian cancer tissue. MATERIALS: Histologically proven primary ovarian cancer samples (n=20) were obtained from women undergoing gynecologic surgery. Frozen sections of ovarian cancer tissues were evaluated by means of indirect immunofluorescence. RESULTS: The expression of hCG protein was ubiquitous throughout all samples, while LH/hCGR was detected in only sixty percent of samples. Co-expression of LH/hCGR and hCG protein was detected in some individual cells in tumor tissue. Some cancer cells expressed only hCG protein, but not LH/hCGR. CONCLUSIONS: Expression of LH/hCG receptor is a characteristic feature of various histological types of ovarian cancer. Co-expression of LH/hCGR and hCG protein in individual cancer cells may point to an autocrine or paracrine mechanism of _hCG activity in the tumor microenvironment. Further studies are needed to evaluate LH/hCGR and hCG protein function in the course of neoplastic development.

The role of repeat uterine evacuation in the management of persistent gestational trophoblastic disease.

OBJECTIVE: To evaluate the role of second (and third) uterine evacuation in the management of persistent gestational trophoblastic disease (GTD). METHODS: This was an observational study of all cases registered over a 10-year period at the Trophoblastic Disease Centre at Weston Park Hospital, Sheffield. Five hundred and forty-four of 4050 women registered during 1991-2000 underwent a second uterine evacuation following a presumptive diagnosis of persistent GTD. The reason for evacuation, hCG level prior to the procedure, histological appearances of evacuated products and the clinical outcome (in terms of the need for chemotherapy) were determined. RESULTS: After a second uterine evacuation 368 patients (68%) completed the follow-up programme without further evidence of persistent disease or need for chemotherapy. If the diagnosis of persistent GTD was confirmed solely on the basis of elevated hCG levels then 171 of 282 (60%) patients did not require chemotherapy. Chemotherapy was more likely where there was histological evidence of persistent trophoblastic disease and where the urinary hCG was >1500 IU/L at the time of the repeat evacuation. Twenty-eight of 60 patients (46%) undergoing a third evacuation required chemotherapy. CONCLUSION: Second uterine evacuation can be a useful therapeutic option for patients with presumed persistent trophoblastic disease not mandating immediate chemotherapy, particularly where the hCG level is <1500 IU/L. Patients with documented persistent trophoblastic disease on histological examination of the second evacuation sample are more likely to require chemotherapy. Third evacuation is not now recommended.

[Choriogonadotropin measurements--critical assessment of new diagnostic possibilities]. / Choriogonadotropina--krytyczna ocena nowych mozliwosci wykonywania oznaczen hormonu w diagnostyce medycznej.

In the human blood and urine there are different antigen forms of choriogonadotropin including nonnicked hCG, nicked hCG, free subunit, free subunit, nicked free, core fragment hCG, hyperglycosylated hCG, nicked hCG missing the subunit C-terminal peptide, asialo hCG, residues 92-145, alternatively nicked hCG--cleaved at 43-44 or 44-45 and others. The authors discuss the value of hCG measurements in the diagnosis of normal pregnancy, pathological pregnancy, Down syndrome screening and oncology: gestational trophoblastic disease, testicular, bladder, digestive tract and other cancers. Special consideration is given to false positive values--phantom hCG--and the consequences of needless therapy.

Ectopic pregnancy: histopathology and assessment of cell proliferation with and without methotrexate treatment.

OBJECTIVE: To evaluate tubal morphology, trophoblast proliferation, and inflammatory reaction in response to methotrexate (MTX) treatment of ectopic pregnancy (EP). DESIGN: Nonrandomized controlled study. SETTING: Academic hospital. PATIENTS: Archival specimens from 10 EP unsuccessfully treated with MTX and 10 cases primarily treated by surgery. INTERVENTIONS: Ki67/hCG and Ki67/human placental lactogen double immunohistochemical methods were used to examine trophoblastic spread, placentation, hormone production, decidualization, vascular invasion, hemorrhage, rupture, and proliferative index of the cytotrophoblast. B and T-lymphocyte responses were evaluated by CD3 and CD20. RESULTS: Trophoblastic spread and placentation were confined to the tubal mucosa after MTX treatment, whereas invasion of the muscularis and subserosa was common in the controls. The proliferative index was reduced (19 percent versus 93 percent), although a high proliferative index was found in two of three cases complicated by rupture. Polar proliferation of Ki67-positive cytotrophoblast toward the implantation site was abolished in MTX-treated cases. Decidual reaction was not observed. No correlation was observed between the above-mentioned findings and gestational age, level of beta-hCG, dose of MTX, or interval to surgery. CONCLUSION: Trophoblastic spread, differentiation, and invasion were compromised by MTX treatment. Methotrexate seems to decrease cytotrophoblast proliferation. Whether a missing decrease in proliferation index reflects treatment failure awaits a larger population-based study.

In-vivo and in-vitro assessment of the influence of ritodrine and oxytocin on the placental secretion of human chorionic gonadotrophin and placental lactogen.

The purpose of this study was to evaluate, in vivo and in vitro, the influence of ritodrine and oxytocin on the placental release of human chorionic gonadotrophin (HCG) and placental lactogen (HPL). The in-vivo study was performed on maternal sera collected before and 1 h after the onset of either ritodrine treatment (50 micrograms i.v./min; administered to 15 women at risk of premature labour) or oxytocin infusion (2 mU i.v./min; administered to 21 women for acceleration of slow labour). The in-vitro study was performed on human term placental explants incubated in the presence of 4-400 ng ritodrine/ml or 15-1500 microU oxytocin/ml. HCG and HPL were measured by radioimmunoassay on maternal sera and incubation media. Maternal circulating concentrations of HCG and HPL remained unaffected after 1 h of ritodrine or oxytocin treatment. The in-vitro release of HCG and HPL by placental explants was not modified when ritodrine or oxytocin was added to the incubation media. The lack of influence of ritodrine and oxytocin on the placental secretion of HCG and HPL suggests that beta 2-adrenergic and oxytocin receptors are not involved in the releasing process.

Valoración de alfa fetoproteína y gonadotropina coriónica en el diagnóstico de 290 tumores germinales testiculares / Evaluation of alpha-fetoprotein and chorionic-gonadotropin in the diagnosis of 290 testicular germinal tumors

Desde el punto de vista terapéutico y pronóstico, los tumores germinales de testículo se dividen en seminoma puro y no seminoma. El diagnóstico definitivo toma en cuenta la histología, la presentación clínica, y los niveles séricos de alfa fetoproteína (AFP) y fracción beta de gonadotropina coriónica humana (HGC). Objetivo: Correlacionar los valores de AFP y HGC con la variedad histológica y etapa clínica para determinar si cambia el diagnóstico final del tumor. Material y métodos: Se tomó suero en la valoración inicial de 290 pacientes con tumores germinales testiculares determinando niveles de AFP y HGC (ELISA, Quantum II de Abbott). Se revisaron los expedientes para conocer estirpe histológica y estadio clínico. Se consideraron seminomas los tumores con histología de seminoma puro sin elevación de AFP y HGC menor de 500 mUI/ml; los tumores germinales no seminomatosos de testículo (TGNST) presentaron histología de no seminoma o de seminoma con elevación de AFP o HGC mayor de 500 mUI/ml. Resultados: Histológicamente se encontraron 120 seminomas puros y 170 TG-NST. Se reclasificó a 13 casos de seminomas como TGNST ya que 10 presentaron elevación de AFP (tres AFP únicamente y siete AFP más HGC) y tres casos tuvieron elevación de HGC > 500 mUI/ml. La distribución final fue de 107 seminomas y 83 seminomas. Conclusiones: Al analizar los niveles séricos de AFP y HGC con la histopatología se obtiene el diagnóstico definitivo de la estirpe tumoral testicular mejorando la selección del tratamiento

Embryonic human chorionic gonadotropin secretion and hatching: poor correlation with cleavage rate and morphological assessment during preimplantation development in vitro.

Of 593 bipronucleate eggs allowed to develop in vitro, 275 (46%) achieved the blastocyst stage and beyond, 124 (21%) initiated hatching, but only 49 (8%) fully hatched. About half of the pre-embryos (48%) which developed to these more advanced stages were incapable of secreting significant amounts (> 200 microIU) of cumulative human chorionic gonadotropin (HCG) up to day 14. HCG production does not appear to begin until the expanded stage and is independent of hatching. Assessing cleavage rate through successive stages and morphological grades up to the 8-cell stage had little bearing on the ability of a pre-embryo to hatch or secrete HCG. Progression through the stages of preimplantation development in vitro does not always appear to be accompanied by the necessary biochemical stages. If only 46% of pre-embryos with two pronuclei are capable of achieving the blastocyst stage, and of these only 52% are capable of secreting HCG, then it follows that only 24% of the original bipronucleate pre-embryos in vitro can be considered anatomically and biochemically competent. However, this is only applicable for pre-embryos not transferred or frozen, and is thus subject to a selection bias. Inability to detect HCG in vitro is not conclusive proof that a pre-embryo is developmentally incompetent. Similarly failure to hatch in vitro may not be taken as definitive evidence that hatching would have failed had fertilization and development been completed in vivo. Nevertheless, if pre-embryonic development in vitro is similar to that in vivo, this may be a contributory factor in the low pregnancy rates following in-vitro fertilization treatment.

Disparity between beta-core levels in pregnancy urine and serum: implications for the origin of urinary beta-core.

We used a highly purified preparation of the naturally occurring core fragment of hCG beta (beta-core) and a new RIA for beta-core to investigate the concentrations and behavior of beta-core in serum and urine. We collected serum and 24-h urine specimens from healthy pregnant women during the first trimester of pregnancy. The concentrations of beta-core in serum were determined by analysis of fractions eluted from Sephadex G-100. Serum concentrations of beta-core immunoreactivity were very low (0.13-1.25 micrograms/L), while large amounts of beta-core were excreted in urine during pregnancy (as much as 4-5 mg/day). Interference with measurement by serum factors did not account for the low levels of beta-core immunoreactivity in pregnancy serum. Based on the known urinary clearance rate of beta-core in healthy nonpregnant subjects, we calculated that urinary clearance of serum beta-core accounts for only about 1% of the beta-core in pregnancy urine. We conclude that during pregnancy, the concentrations of beta-core in plasma are measurable, but extremely low, and that most of the beta-core in urine is derived by mechanisms other than urinary clearance of serum beta-core; most likely, these mechanisms involve nephrogenous production of beta-core from precursor molecules such as hCG and hCG beta.

A gonadotropin and alpha-subunit suppression test for the assessment of the ectopic production of human chorionic gonadotropin and its subunits after the menopause.

One contraceptive tablet [(CP) containing 0.05 mg ethinyl estradiol and 0.5 mg norgestrel] was administered daily for 7 days to 38 hypergonadotropic postmenopausal women who had benign illness and to 34 similar women with cancer, in order to suppress the gonadotropins (LH and FSH) and their alpha-subunit secreted by the pituitary. LH, FSH, alpha-subunit, and hCG were measured using RIA in serum and urine extracts obtained before and on the seventh day of treatment. In the control group serum alpha-subunit fell to 53%, serum LH to 51%, serum FSH to 36%, urine alpha to 42%, urine hCG to 54%, urine LH to 53%, and urine FSH to 44% of their mean pretreatment values. Serum hCG was undetectable after treatment in all except 11 women in whom serum LH was not adequately suppressed. This fact and the significant positive linear correlation found between basal serum hCG and LH imply that serum hCG measured in postmenopausal women, with a RIA using the SB6 antiserum, is mainly cross-reacting LH. The suppression proved to be useful in the assessment of ectopic production of hCG and its alpha-subunit in the cancer group. Before treatment, 3 patients (8.8%) had elevated serum alpha-subunit levels, whereas after treatment 9 patients (26.5%) had elevated levels. For urine alpha-subunit 3 patients (8.8%) had elevated levels before and 13 (38.2%) after treatment. Six patients (17.7%) had elevated serum hCG levels before and 7 (20.6%) after treatment; however, for urine hCG, 3 patients (9%) had elevated levels before and 8 (24.2%) after treatment. Sephadex G-100 chromatography of urine extracts from two control women showed that the predominant form of free alpha-subunit secreted by the pituitary and excreted in the urine had lower apparent molecular weight after treatment with the CP. Chromatography of urine extracts from two cancer patients demonstrated that a small amount of beta-subunit of hCG produced ectopically by the tumor and excreted into the urine along with a core-type fragment of the hCG-beta was masked in the basal urine by cross-reacting LH; these ectopic peptides were not suppressible and could be specifically measured only after elimination of the urine LH by treatment with the CP. We conclude that administration of contraceptive steroid to post-menopausal women with cancer, by suppressing pituitary gonadotropin secretion and thus minimizing their interference in the hCG RIAs, can be useful in the detection of ectopic production of the hCG and its subunits.

An assessment of Leydig cell function after bilateral or unilateral efferent duct ligation: further evidence for local control of Leydig cell function.

Leydig cell function was examined in adult male rats at 2 and 4 weeks after bilateral or unilateral efferent duct ligation. Bilateral efferent duct ligation resulted in significantly elevated serum LH levels, an increased size of Leydig cells and an enhanced testosterone response to gonadotropin stimulation in vitro despite a marked loss of LH-hCG receptors. After unilateral ligation of the vasa efferentia, the parameters of Leydig cell function in the ligated testes showed identical changes to those seen after bilateral ligation, whereas such changes were minor or absent in the unligated contralateral testes of the same animals. These results demonstrate that the modifications of Leydig cell function after efferent duct ligation are mainly due to local changes within the testes providing further evidence for an intratesticular control of Leydig cell function.

Brain metastases in malignant teratoma: a review of four years' experience and an assessment of the role of tumour markers.

Between 1973 and 1977, 247 patients with malignant teratoma have been treated in two units in London. Seventeen have developed brain metastases, an overall incidence of 6.2%. The median survival from diagnosis of cerebral metastases is 6 weeks and all patients except one have died. The survivor is disease-free 12 months after completing treatment, which included extensive use of chemotherapy, surgery and radiotherapy. Serum gonadotrophin (HCG) and alpha-foetoprotein (AFP) estimations have been performed in 264 patients as a means of monitoring the effects of therapy. In 42 patients (37 of whom had Stage IV disease) the peak HCG level was greater than 10(4) iu/l, and the incidence of brain metastases in this group was 26%, significantly higher than in the group with HCG levels below 10(4) iu/l, for which the incidence of cerebral deposits was 1.8% (P less than 0.0001). No significant correlation was seen between peak AFP levels and the incidence of brain metastasis. With the aim of improving results by earlier diagnosis, cerebrospinal fluid (CSF) specimens have been examined for HCG and AFP levels in 56 subjects, 9 of whom had brain metastases. A serum: CSF HCG ratio less than 40 is an accurate indication of the presence of brain metastases, and may have considerable predictive value. However, false-negative serum: CSF HCG rations (greater than 40) frequently occur in patients with proven brain deposits. Estimation of AFP in spinal fluid has not contributed to the early diagnosis of brain metastases in malignant teratoma.