Testicular seminoma with human chorionic gonadotropin production

Testicular seminoma with elevated serum human chorionic gonadotropin level (hCG-positive seminoma) is regarded as more malignant than marker-negative seminoma, although its prognosis is still unclear. To clarify the malignant potential of seminoma with hCG production, the serum levels of the beta subunit of hCG (beta-hCG) and lactic acid dehydrogenase (LDH) were examined in 35 and 40 patients, respectively, and the immunohistochemical expression of beta-hCG examined in 45 tumors. The elevation of the LDH serum level correlated to the invasive status, metastatic status and poor outcome, while that of the serum beta-hCG level correlated only to the metastatic status. Immunohistochemical expression of beta-hCG was observed in syncytiotrophoblastic giant cells in 11 tumors and a few mononuclear seminoma cells in 36 tumors. Expression was not associated with the malignancy potential, except where the expression in mononuclear cells inversely correlated to the invasive status. These results suggest that most seminomas produce a slight amount of hCG; that an elevated hCG serum level indicates the presence of metastatic tumors and mainly reflects an increase in tumor volume but not in cellular malignancy potential; and that the LDH serum level, rather than hCG, is more useful as a prognostic indicator for patients with seminoma.     

Free alpha-subunits of human chorionic gonadotropin in preeclampsia

OBJECTIVE: To investigate free alpha-human chorionic gonadotropin (hCG) as a marker of preeclampsia. METHODS: Four groups of patients were studied: normal pregnancies, preeclampsia, eclampsia and normal pregnant women <20 weeks' gestation. Patients were further divided according to parity and gestational age (< or =20, 21-30, 31-40 weeks). An immunoradiometric assay employing monoclonal antibodies specific for free alpha-hCG was used. RESULTS: A total of 313 patients were analyzed. Thirty-four patients < or =20 weeks' gestation were followed until delivery: five (14.7%) developed preeclampsia; none had abnormal alpha-hCG levels before onset of preeclampsia. Patients with preeclampsia (21-30 weeks' gestation) demonstrated a mean alpha-hCG level greater than that of normotensive controls but this was not statistically significant. Between 31 and 40 weeks' gestation, mean alpha-hCG levels in the hypertensive and control groups were 210.8 ng/ml and 115.8 ng/ml, respectively (P < 0.001). A stronger association was observed between alpha-hCG and preeclampsia with increasing gestational age (relative risk [RR] 2.07, 21-30 weeks; RR 3.02, 31-40 weeks) and severity (RR 4.51, mild; RR 12.15, severe; RR 16.88, eclampsia). CONCLUSION: There is a strong association between alpha-hCG and preeclampsia, nevertheless this test is unsuitable for predicting preeclampsia.     

Highly specific enzyme immunoassay for human chorionic gonadotropin

A highly specific enzyme immunoassay for determining hCG was established by using beta-D-galactosidase as label. In order to increase the specificity of the assay, an antiserum against whole hCG was purified on a column of hCG beta carboxyl-terminal peptide (residues 123-145) covalently linked to Sepharose 4B. The antibody (N101S) thus prepared showed a weak cross-reactivity with human LH in an assay using hCG-enzyme conjugate, but the slight cross-reactivity was virtually avoided when an hCG beta carboxyl-terminal peptide was used as a peptide in the enzyme conjugate. N101S antibody was compared with antiserum (B1B) directed against a carboxyl-terminal peptide (123-145). In hCG measurement N101S gave about 30 times higher sensitivity than B1B, although the former antibody was less sensitive to carboxyl-terminal peptides of hCG beta. The enzyme immunoassay using a combination of N101S antibody and a carboxyl-terminal peptide (130-145)-enzyme conjugate was able to detect as little as 0.25 mIU of hCG without the interference of LH. The performance and validity of this assay were comparable to those of conventional radioimmunoassay.     

Mechanisms of chorionic gonadotropin action on splenocytes, suppressed in their humoral immune response

Chorionic gonadotropin (CG) used at 10 and 50 IU/ml concentrations was introduced into a short-term macroculture of splenocytes of CBA mice suppressors fractionated on a nylon wool column. Regulatory effects of CG were evaluated in a syngeneic transfer system by the degree of inhibition of a humoral immune response in recipients. It was found that CG (50 IU/ml) inhibits the functional activity of splenocyte suppressors. The observed effect of the hormone was blocked by introduction into the incubation medium of the inhibitor, inositol-1-monophosphatase (lithium), and did not depend on cyclooxygenase (voltaren) or potential-independent Ca(2+)-channel (verapamil) inhibitors. Determination of the intracellular cAMP level by radioimmunoassay revealed a reliable increase of this secondary messenger in splenocyte suppressors during their incubation with CG. The ability of CG (50 IU/ml) to inhibit the functional activity of intact T-lymphocytes (but not B-cells) was established. The suppressive hormone activity was also followed by cAMP level increase in target cells. These findings suggest that the mechanism of a transmembrane transfer of the CG signal in splenocyte suppressors is related to the adenylate cyclase system and, probably, to phosphoinositide metabolism products.     

Regulation of chorionic gonadotropin gene expression

This paper describes the development of a high-performance liquid chromatographic method for the quantitation of free carnitine, total carnitine, acetylcarnitine, propionylcarnitine, isovalerylcarnitine, hexanoylcarnitine and octanoylcarnitine in human urine. Carnitine and acylcarnitines were isolated from 10 or 25 microliters of urine using 0.5-ml columns of silica gel, derivatized with 4'-bromophenacyl trifluoromethanesulfonate and separated by high-performance liquid chromatography. Using 4-(N,N-dimethyl-N-ethylammonio)-3-hydroxybutanoate ("e-carnitine") as the internal standard, standard curves (10-300 nmol/ml) were generated. Carnitine and acylcarnitines were quantified (when they were present) in normal human urine and the urine of patients diagnosed with one of three different disorders of organic acid metabolism: methylmalonic aciduria, isovaleric aciduria, isovaleric acidemia, and medium-chain acyl-CoA dehydrogenase deficiency.     

Culture of first-trimester and full-term human chorionic villus explants: role of human chorionic gonadotropin and human placental lactogen as a viability index

In long-term cultures of human chorionic villus explants, the viability of the tissue must be controlled to ensure the reliability of functional studies. Ionic levels (pH), gas concentrations (pO2, pCO2) and metabolic markers (glucose, lactate) in the culture medium are often utilized. Analyses of hormone, enzyme and protein levels are also frequently used to estimate viability. The purpose of this study was to evaluate whether in vitro release and immunoreactivity of human chorionic gonadotropin (hCG) and human placental lactogen (hPL) were correlated with the viability of first-trimester and full-term chorionic villus explants as determined by histopathology. Villus explants of first-trimester and full-term pregnancies were incubated in 6-well plates of RPMI medium which was supplemented with 10% fetal calf serum. Incubations were performed for 10 days, and the plates were kept at 37 degrees C under a water-saturated atmosphere containing 5% CO2 and 95% O2. The medium was replaced every day and samples of supernatant were frozen for later testing of hCG (first trimester) or hPL (full term), glucose consumption and lactate production. The tissue was also fixed and embedded for light-microscopic examination and immunocytochemistry. The hCG release remained stable during 6-7 days at a high level before decreasing, whereas hPL release decreased during the first 5-6 days then stabilized at a relatively low level. Only hCG kinetics were significantly different between tissue incubated with and without cycloheximide or iodoacetic acid. Both hCG and hPL immunoreactivity were not significantly different between tissue cultures with, and without, addition of cycloheximide or iodoacetic acid and even with morphological evidence of trophoblast and endothelial necrosis. The immunoreactivity for both hormones remains highly positive when the significant release has stopped, and does not reflect the tissue viability.     

Cytomorphological changes in the rabbit oviductal epithelium after human chorionic gonadotropin treatment

Progenitor cells were isolated from the developing human central nervous system (CNS), induced to divide using a combination of epidermal growth factor and fibroblast growth factor-2, and then transplanted into the striatum of adult rats with unilateral dopaminergic lesions. Large grafts were found at 2 weeks survival which contained many undifferentiated cells, some of which were migrating into the host striatum. However, by 20 weeks survival, only a thin strip of cells remained at the graft core while a large number of migrating astrocytes labeled with a human-specific antibody could be seen throughout the striatum. Fully differentiated graft-derived neurons, also labeled with a human-specific antibody, were seen close to the transplant site in some animals. A number of these neurons expressed tyrosine hydroxylase and were sufficient to partially ameliorate lesion-induced behavioral deficits in two animals. These results show that expanded populations of human CNS progenitor cells maintained in a proliferative state in culture can migrate and differentiate into both neurons and astrocytes following intracerebral grafting. As such these cells may have potential for development as an alternative source of tissue for neural transplantation in degenerative diseases.     

Immunoprocedures for detecting human chorionic gonadotropin: clinical aspects and doping control

The pregnancy hormone human chorionic gonadotropin (hCG) is also present at low concentrations in plasma and urine of men and nonpregnant women. hCG immunoreactivity occurs in various molecular forms: Besides the intact hCG heterodimer, considerable amounts of proteolytically cleaved forms, free subunits, and fragments are found in plasma and urine. Especially in urine, proteolytic fragments constitute a major part of the hCG immunoreactivity. The different forms of hCG cross-react to various degrees in immunoassays and constitute a problem for standardization of specific hCG determinations. After injection of hCG (10,000 IU of Pregnyl; Organon), above-normal concentrations of hCG can be detected in serum and urine for 7-11 days. Most immunoassays for hCG also measure hCG beta. Quantitative hCG determinations are mainly performed on serum samples, and very few commercial hCG determinations have been validated for determination of urine samples. Considerable care must therefore be exercised when utilizing such assays to analyze urines for doping control.     

Mass spectrometric characterization of nicked fragments of the beta-subunit of human chorionic gonadotropin

To test the hypothesis that hypoglycemia unawareness and impaired counterregulation are reversible after meticulous prevention of hypoglycemia in IDDM patients with diabetic autonomic neuropathy (DAN), 21 patients (8 without DAN [DAN-]; 13 with DAN [DAN+]; of the latter, 7 had orthostatic hypotension [DAN+PH+] and 6 did not [DAN+PH-]) and 15 nondiabetic subjects were studied during stepped hypoglycemia (plateau plasma glucose decrements from 5.0 to 2.2 mmol/l) before and 6 months after prevention of hypoglycemia (intensive therapy). After 6 months, frequency of mild hypoglycemia decreased from approximately 20 to approximately 2 episodes/patient-month while HbA1c increased from 6.2 +/- 0.3 to 6.9 +/- 0.2% (P < 0.05). Responses of adrenaline improved more in DAN- patients (from 1.17 +/- 0.12 to 2.4 +/- 0.22 nmol/l) than in DAN+PH- (from 0.75 +/- 0.25 to 1.56 +/- 0.23 nmol/l) and DAN+PH+ patients (from 0.80 +/- 0.24 to 1.15 +/- 0.27 nmol/l, P < 0.05) but remained lower than in nondiabetic subjects (4.9 +/- 0.37 nmol/l, P < 0.05), whereas glycemic thresholds normalized only in DAN-, not DAN+. Autonomic symptoms of hypoglycemia improved but remained lower in DAN- (6.2 +/- 0.6) than in nondiabetic subjects (8.1 +/- 1.1) and lower in DAN+PH+ (4 +/- 0.8) than in DAN+PH- subjects (5.1 +/- 0.8, P < 0.05), whereas neuroglycopenic symptoms normalized (NS). Cognitive function deteriorated less before than after prevention of hypoglycemia (P < 0.05). Thus, intensive therapy with emphasis on preventing hypoglycemia reverses hypoglycemia unawareness in DAN+ patients despite marginal improvement of adrenaline responses, results in low frequency of hypoglycemia despite impaired counterregulation, and maintains HbA1c in the range of intensive therapy. We conclude that DAN, long IDDM duration per se, and antecedent recent hypoglycemia contribute to different extents to impaired adrenaline responses and hypoglycemia unawareness.     

Polypeptide pattern of human breast epithelial cells following human chorionic gonadotropin (hCG) treatment

Numerous attempts have made to describe the particular protein pattern of malignant cells by using high resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). The placental hormone human chorionic gonadotropin (hCG) inhibits tumor initiation and progression in experimental animals and has an inhibitory effect on the proliferation of human breast epithelial cells (HBEC) in vitro. The inhibitory effect on the immortalized HBEC MCF-10F is accompanied by the immunocytochemical expression of inhibin alpha and beta subunits by treated cells. With the purpose of clarifying the molecular mechanisms involved in this effect, the pattern of protein synthesis and mRNA were studied by 2-D PAGE in the immortalized HBEC MCF-10F cells treated in vitro 1001U for 24 h. The effect of hCG treatment on the synthesis of MCF-10F cells was monitored by labeling both control and treated cells with [S35]methionine and separation by 2-D PAGE. At least 11 proteins were preferentially synthesized and five specific polypeptides were decreased in hCG treated cells in comparison with controls. The hCG induced at least four new mRNAs which encoded protein in the molecular mass range of 24-72 kDa. It also increased the expression of at least six mRNAs and reduced the expression of least four mRNAs in comparison with control cells. The hCG-treated cells actively synthesized a 33-kDa polypeptide which was not present in control cells. The nature of this hCG-inducible 33 kDa protein elucidated by immunoprecipating [S35]methionine-labeled proteins with antisera directed against rat inhibin subunit alpha and beta b.(ABSTRACT TRUNCATED AT 250 WORDS)     

The effect of human chorionic gonadotropin on methotrexate concentration in the rat testes

PURPOSE: We analyzed the effect of human chorionic gonadotropin (hCG) on drug concentrations in testicular interstitial fluid and whole testis tissue samples in rats receiving hCG prior to methotrexate (MTX) administration and in animals that did not receive hCG. MATERIALS AND METHODS: Adult male Sprague-Dawley rats were injected subcutaneously with 200 i.u. hCG (Goldline Laboratories, Ft. Lauderdale, FL.). Controls were injected subcutaneously with normal saline (0.2 cc). Sixteen hours after injection, each rat was given methotrexate (Methotrexate LPF, Immunex Corp. Seattle WA.) via a carotid artery cannula in a dose of 30 mg./kg. Methotrexate (MTX) levels were collected 60 minutes post infusion time in 27 rats and 90 minutes post infusion in 27 rats. MTX levels were measured in serum, testicular interstitial fluid and testicular tissue. MTX levels were measured using high performance liquid chromatography (HPLC). RESULTS: A significantly higher concentration of MTX was found in testicular interstitial fluid (TIF) in rats injected with hCG when specimens were collected 60 minutes post infusion. MTX levels in TIF had reversed 90 minutes post infusion with higher levels found in control rats. Tissue levels of MTX demonstrated no significant difference at either 60 or 90 minutes in the hCG treated animals or controls. CONCLUSION: Our results suggest that hCG effects the tissue distribution of MTX within the testis. Human chorionic gonadotropin may have this effect on the testicular microvasculature by 1) selectively increasing capillary permeability, 2) increasing lymphatic flow within the testes or 3) increasing testicular blood flow.     

Choriocarcinoma with hyperthyroidism: probable identity of the thyrotropin with human chorionic gonadotropin

A 15-year-old girl developed severe hyperthyroidism secondary to metastatic choriocarcinoma. Her serum contained high levels of human chorionic gonadotropin (HCG) by radioimmunoassay and had a thyroid-stimulating activity different from that of pituitary thyrotropin (TSH) or of long-acting thyroid stimulator (LATS) in the McKenzie mouse bio-assay. Gradient ultracentrifugation localized this thyroid stimulator to a narrow zone midway between markers of transferrin and ovalbumin. On gel filtration it emerged just before albumin as a single peak coinciding with the peak for HCG. Her HCG was identical to an authentic sample of HCG in position to gel filtration columns and on gel electrophoresis. These results suggest that the thyrotropin of choriocarcinoma is HCG.     

Beta-core fragment of human chorionic gonadotropin in cervical intraepithelial neoplasia (CIN)

The effect of inhalation of paprine dust on the bronchopulmonary tract and antibody-forming function of the spleen was studied experimentally on two lines of mice: CBA and CC57W, differing from each other by haplotype H-2 and antioxidant status of the organisms under study. During comparative histomorphological examination of lungs and bronchi of the experimental animals a more intensive reaction was found in mice of the CBA line, whereas in mice of the CC57W line vascular changes prevailed without any reactions in the bronchioles. Reactions of the spleen to paprine dust in animals of different lines were also different: the quality of antibody-forming cells in the spleen of CBA line mice with comparatively higher antioxidant status increased under experimental conditions, while it decreased in those of the CC57W line. The data obtained showed dependance of the sensitivity of the organism to action of operational dust on individual genetic properties, demonstrating thus the necessity of applying principles of medical genetics in order to prevent effectively the development of occupational diseases in exposed teams of workers.     

The effect of preparations of human chorionic gonadotropin on lymphocyte stimulation and immune response

Frontal sharp waves (encoches pointues frontales) identified by Monod (1960) were studied in 110 neonates and infants. Three pattern were described: typical, related and degraded. They were recorded in all subjects, the reason why the frequency of each pattern varied was discussed. Pathological states did not increase the occurrence of those frontal sharp waves since most of them were recorded in children either normal or suffering from a minor pathology. Incidence of maturation seemed to be probable since the highest proportion of frontal sharp waves occurred in infants whose gestational and legal age were respectively greater than 43 weeks and between 20 and 40 days at the time of investigation. Typical and degraded forms were predominant in transitional period towards quiet sleep; they were less numerous during falling asleep in active sleep and totally absent during active sleep following quiet sleep. This could suggest that the two states of active sleep are different in nature.     

Characterization of monoclonal antibodies recognizing alpha and beta subunits of human chorionic gonadotropin hormone

The purpose of this study was to characterize the pharmacological effects of 2-[[4-(o-methoxyphenyl)piperazin-1-yl]methyl]-1,3-dioxoperhydro imidazo[1,5-a]pyridine (B-20991) by using several biochemical and behavioral assays. Results of binding studies showed that B-20991 binds with high affinity to the 5-HT1A receptor (Ki = 31.7 +/- 1.7 nM), moderate affinity to 5-HT3 receptor (Ki = 269.4 +/- 23.2 nM) and low affinity (Ki > 1000) to 5-HT2A receptor, dopamine D2 receptor, benzodiazepine receptors and alpha1-adrenoceptor. The administration of B-20991 produced a dose and time related decrease in mouse rectal temperature, increased both lower lip retraction and flat body posture behavioral scores in rat, decreased 5-hydroxytryptamine (5-HT, serotonin) neuronal activity in mouse hypothalamus, and did not alter dopamine neuronal activity nor locomotor activity. The anxiolytic activity of B-20991 was assessed by using both the social interaction and light/dark box tests. The results of these tests indicated that B-20991 caused a dose-related increase in the social interaction and light/dark box behavioral scores. Taken together, these results suggest that B-20991 is a 5-HT1A receptor agonist that exhibits anxiolytic activity.     

Bovine cyclic endometrium contains high-affinity luteinizing hormone/human chorionic gonadotropin binding sites

High-affinity LH/hCG binding sites have been characterized in porcine, lepine, and murine uteri. In the present study, LH/hCG binding sites were characterized in bovine endometrium. Radioreceptor assays were performed with membrane homogenates of endometrial tissues and analyzed for binding site specificity and capacity. There was little competition for receptor occupancy between hCG and ovine FSH (5%) or ovine prolactin (< 0.1%), but there was a 20% cross-reaction with eCG. There was no affinity for LH/hCG in crude membrane preparations of kidney, skeletal muscle, or vascular tissues. Concentrations of endometrial LH/hCG binding sites were determined during the bovine estrous cycle. LH/hCG receptors were found in cell preparations from Days 2-4 and 15-17 of the cycle, but not in preparations from the other stages of the cycle tested (Days 8-12, pre- and post-estrus, and ovulation). The concentration of uterine LH/hCG receptor varied during the estrous cycle, with higher values at Days 15-17 (3.1 fmol/mg protein) and lower values at Days 2-4 (1.2 fmol/mg protein). However, the binding capacity of hCG by luteal cells (9.7 fmol/mg protein) was 3-fold higher (p < 0.01) than that by endometrial tissue on any day studied. No differences in affinity constant (Ka) were seen between endometrial LH/hCG receptors (either) from Days 2-4 or 15-17) and mid-cycle luteal cells (0.60 x 10(11) M-1). Using Western blot analysis, we determined the expression of cyclooxygenase (COX) during the estrous cycle of the cow. It was found that the signal for COX was strongest at 15-17 days.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of human chorionic gonadotropin on pregnancy rates in lactating dairy and beef cows

Single subcutaneous injection of 1500 IU human chorionic gonadotropin or placebo injections were given at the time of insemination in lactating dairy and beef cows to determine their effects on rates of pregnancy. Pregnancy rates at first service in 161 control and 145 treated dairy cows were 52.8% and 44.8%, respectively. Similar rates for 136 control and 145 treated beef cows were 54.4% and 54.5%. Injections of human chorionic gonadotropin were not effective in stimulating conception rates in lactating dairy or beer cows under field conditions.