In vitro differential responses of rat and human aryl hydrocarbon receptor to two distinct ligands and to different polyphenols.

TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) and several other environment/food-borne toxic compounds induce their toxicity via the aryl hydrocarbon receptor (AhR). AhR is also modulated by various endogenous ligands e.g. highly potent tryptophan (Trp)-derivative FICZ (6-formylindolo[3,2-b]carbazole) and natural ligands abundant in the human diet e.g. polyphenols. Therefore, evaluating AhR species-specific responses is crucial for understanding AhR physiological functions, establishing risk assessments, and exploring the applicability of AhR mediators in drug and food industry towards human-based usages. We studied AhR transactivation of FICZ/TCDD in vitro in a time-dependent and species-specific manner using dioxin responsive luciferase reporter gene assays derived from rat (DR-H4IIE) and human (DR-HepG2) hepatoma cells. We observed for the first time that FICZ potency was similar in both cell lines and was 40 times higher than TCDD in DR-HepG2 cells. Depleting Trp-derivative endogenously produced ligands by using culture medium without Trp, resulted in 3-fold higher AhR activation upon adding FICZ in DR-H4IIE cells, in contrast to DR-HepG2 cells which revealed a fast degradation of FICZ induction from 10 h post-exposure to complete disappearance after 24 h. Seven polyphenols and a mixture thereof, chosen based on commercially recommended doses and adjusted to human realistic exposure, caused rat and human species-specific AhR responses. Two isoflavones (daidzein and genistein) induced rat AhR synergistic effects with FICZ and/or TCDD, while quercetin, chrysin, curcumin, resveratrol, and the mixture exerted a strong inhibitory effect on the human AhR. Strikingly, resveratrol and quercetin at their realistic nanomolar concentrations acted additively in the mixture to abolish human AhR activation induced by various TCDD concentrations. Taken together, these results illustrate the species-specific complexity of AhR transcriptional activities modulated by various ligands and highlight the need for studies of human-based approaches.

Elucidating the aryl hydrocarbon receptor antagonism from a chemical-structural perspective.

The aryl hydrocarbon receptor (AhR) plays an important role in several biological processes such as reproduction, immunity and homoeostasis. However, little is known on the chemical-structural and physicochemical features that influence the activity of AhR antagonistic modulators. In the present report, in vitro AhR antagonistic activity evaluations, based on a chemical-activated luciferase gene expression (AhR-CALUX) bioassay, and an extensive literature review were performed with the aim of constructing a structurally diverse database of contaminants and potentially toxic chemicals. Subsequently, QSAR models based on Linear Discriminant Analysis and Logistic Regression, as well as two toxicophoric hypotheses were proposed to model the AhR antagonistic activity of the built dataset. The QSAR models were rigorously validated yielding satisfactory performance for all classification parameters. Likewise, the toxicophoric hypotheses were validated using a diverse set of 350 decoys, demonstrating adequate robustness and predictive power. Chemical interpretations of both the QSAR and toxicophoric models suggested that hydrophobic constraints, the presence of aromatic rings and electron-acceptor moieties are critical for the AhR antagonism. Therefore, it is hoped that the deductions obtained in the present study will contribute to elucidate further on the structural and physicochemical factors influencing the AhR antagonistic activity of chemical compounds.

A mixture of persistent organic pollutants relevant for human exposure inhibits the transactivation activity of the aryl hydrocarbon receptor in vitro.

While humans are exposed to mixtures of persistent organic pollutants (POPs), their risk assessment is usually based on a chemical-by-chemical approach. To assess the health effects associated with mixed exposures, knowledge on mixture toxicity is required. Several POPs are potential ligands of the Aryl hydrocarbon receptor (AhR), which involves in xenobiotic metabolism and controls many biological pathways. This study assesses AhR agonistic and antagonistic activities of 29 POPs individually and in mixtures by using Chemical-Activated LUciferase gene eXpression bioassays with 3 transgenic cell lines (rat hepatoma DR-H4IIE, human hepatoma DR-Hep G2 and human mammary gland carcinoma DR-T47-D). Among the 29 POPs, which were selected based on their abundance in Scandinavian human blood, only 4 exerted AhR agonistic activities, while 16 were AhR antagonists in DR-H4IIE, 5 in DR-Hep G2 and 7 in DR-T47-D when tested individually. The total POP mixture revealed to be AhR antagonistic. It antagonized EC50 TCDD inducing AhR transactivation at a concentration of 125 and 250 and 500 fold blood levels in DR-H4IIE, DR-T47-D and DR-Hep G2, respectively, although each compound was present at these concentrations lower than their LOEC values. Such values could occur in real-life in food contamination incidents or in exposed populations. In DR-H4IIE, the antagonism of the total POP mixture was due to chlorinated compounds and, in particular, to PCB-118 and PCB-138 which caused 90% of the antagonistic activity in the POP mixture. The 16 active AhR antagonists acted additively. Their mixed effect was predicted successfully by concentration addition or generalized concentration addition models, rather than independent action, with only two-fold IC50 underestimation. We also attained good predictions for the full dose-response curve of the antagonistic activity of the total POP mixture.

The use of antibiotics in cattle in North-East Benin: pharmaceutical inventory and risk practices of cattle breeders.

This study's aim is to inventory antibiotics used in cattle in North-East Benin and assess risk practices that could be the cause of both food chain contamination by antibiotic residues and selection of antibiotic-resistant bacteria in animals and humans. A survey was conducted among 98 cattle breeders in the districts of Banikoara, Kandi, Bembereke, and Kalale in North Benin. Semi-structured interviews were conducted, covering breeder status, breeding system, and antibiotic use. Multiple correspondence analysis and hierarchical classification analysis were conducted to establish a breeder typology. Breeders mainly belonged to the Fulani ethnic group (71.4 ± 8.9%) and almost all of them received "no formal education" (96.9 ± 3.4%). Cattle herds were mainly composed of a single breed, the Borgou (76.4 ± 8.1%) or the Fulani Zebu (16.0 ± 7.0%). Some herds were mixed. Antibiotics groups used in cattle breeding were tetracyclines, beta-lactams, sulfonamides, aminoglycosides, and macrolides, used by respectively 100%, 69.4 ± 9.1%, 56.1 ± 9.8%, 44.9 ± 9.8%, and 34.7 ± 9.4% of breeders. These drugs were purchased in local markets (59.0 ± 15.4%) and veterinary pharmacy (41.0 ± 15.4%). They were mainly used against respiratory diseases, lameness, mastitis, omphalitis and neonatal enteritis, and skin diseases. Only 49.0 ± 9.9% of breeders seek veterinary services to treat animals and 92.9 ± 5.1% of them did not respect antibiotic withdrawal times. These practices suggest that both contamination of bovine meat with antibiotic residues and selection of resistant bacteria are to be expected, resulting in adverse health effects on consumers.

Assessment of methods of detection of water estrogenicity for their use as monitoring tools in a process of estrogenicity removal.

Methods of monitoring of estrogenicity in water were gathered, compared, and tested within the context of their practical use as measurement and design tools, in the development of a process of degradation of estrogenic endocrine disruptors. In this work, the focus was put on in vitro assays, with the use of analytical techniques as additional analysis when possible. Practically, from a literature review, four methods that seemed most suitable to practical use required in a process development were tested: the Yeast Estrogen Screen assay, the Lyticase-assisted Yeast Estrogen Screen assay (LYES), the MMV-LUC assay and the HPLC-UV analytical method. Dose-response curves in response to estrogenic standard 17ß-estradiol were compared. Bisphenol A estrogenicity was measured by the methods as well. The model for the calculation of estradiol equivalents as measurements units was adapted. The methods were assessed in terms of ranges of detection, time of experiment, cost, ease of the experiment, reproducibility, etc. Based on that assessment, the LYES assay was selected and successfully applied to the monitoring of estrogenicity removal from 17ß-estradiol and bisphenol A. More precisely, the bioassay allowed the acquisition of kinetic curves for a laboratory-scaled process of estrogenicity removal by immobilized enzymes in a continuous packed-bed reactor. The LYES assay was found to have a real methodological potential for scale-up and design of a treatment process. The HPLC-UV method showed good complementarity with the LYES assay for the monitoring of bisphenol A concentrations in parallel with estrogenicity, reporting no significant estrogenicity from degradation byproducts, among others.

Risk assessment of Belgian adults for furan contamination through the food chain.

Risk assessment is an interdisciplinary process used to quantify the risk linked to a hazard. In the present paper it is applied to quantify the risk linked to furan ingestion through the food chain for the Belgian adult population. Two approaches, deterministic and probabilistic, were carried out in parallel. The deterministic method relied on a case study, whereas the probabilistic approach involved statistical distributions of contamination and consumption data to calculate a statistical distribution of the daily intake. First, the deterministic method revealed a low estimated daily intake (EDI) for the average population (380 ng*(kg(bw)*day)⁻¹) and a huge contribution of coffee consumption to the EDI (55%). Increasing or decreasing the daily coffee consumption by one cup can affect the EDI by about 22%. Afterwards, the probabilistic approach showed that the average population has a low EDI (494 ng*(kg(bw)*day)⁻¹), and that high contamination levels were only registered in a small proportion of the population. Finally, a comparison of the RfD(chronic oral) showed that less than 10% of the Belgian population had an EDI above the reference dose proposed by the USEPA; the majority of the population had an EDI 20% below the reference dose. The margin of exposure (MoE) approach indicated that the level of risk related to furan intake through ingestion is low, with a MoE > 10,000 for more than 10% of the population and no result < 100.

Estimation of furan contamination across the Belgian food chain.

This paper provides an estimate of the furan content of Belgian foods. The objective of the study was to achieve the best food chain coverage with a restricted number of samples (n = 496). The geographic distribution, different market chains and labels, and consumption frequencies were taken into account in the construction of the sampling plan. Weighting factors such as contamination levels, consumption frequency and the diversity of food items were applied to set up the model. The very low detection capabilities (CC(ß)) of the analytical methods used (sub-ppb) allowed reporting of 78.2% of the overall dataset above CC(ß) and, in particular, 96.7% for the baby food category. The highest furan levels were found in powdered roasted bean coffee (1912 µg kg(-1)) with a mean of 756 µg kg(-1) for this category. Prepared meat, pasta and rice, breakfast cereals, soups, and baby food also showed high mean furan contents ranging from 16 to 43 µg kg(-1). Comparisons with contamination surveys carried out in other countries pointed out differences for the same food group and therefore contamination levels are related to the geographical origin of food items.

Potential of an in vitro toolbox combined with exposure data as a first step for the risk assessment of dietary chemical contaminants.

In vitro risk assessment of dietary contaminants has become a priority in human food safety. This paper proposes an in vitro approach associating different complementary tools in an original toolbox and aims to improve the assessment of the toxicological impact of dietary contaminants at realistic human exposure levels, with a special focus on the intestinal compartment. The system is based on the use of four complementary cellular tools, namely stress gene induction in transgenic strains of Escherichia coli, modulation of the activity of key biotransformation enzymes (cytochrome P-450 (CYP) 1A1 and 3A4) in a human intestinal cell line, and activation of aryl hydrocarbon receptor (AhR) and oestrogenic receptor (ER)-dependent genes in agonistic and antagonistic assays with luciferase reporter cells. It was applied to four chosen model molecules: ochratoxin A (OTA) and deoxynivalenol (DON), two common food-borne mycotoxins, and imazalil (IMA) and benomyl (BEN), two fungicides widely occurring in foodstuffs. All these assays were performed at or around a realistic intestinal concentration, determined through a deterministic approach based on the calculation of a theoretical maximum daily intake (TMDI). Using the four model molecules, it is clearly highlighted that induction of CYP1A1 activity and inhibition of CYP3A4 activity occurred in Caco-2 cells at a realistic intestinal concentration of IMA. Furthermore, some bacterial stress genes were induced in a range of realistic concentrations, following exposure to DON and IMA. In addition, BEN clearly provoked an ER agonistic activity in a human oestrogen sensitive reporter cell line. All these results are in accordance with the literature, suggesting that the in vitro toolbox constitutes an interesting approach in order to obtain a first 'fingerprint' of dietary contaminants at realistic human exposure for further risk assessment.

Screening, identification and quantification of glucosinolates in black radish (Raphanus sativus L. niger) based dietary supplements using liquid chromatography coupled with a photodiode array and liquid chromatography-mass spectrometry.

The glucosinolate profile of black radish (Raphanus sativus L. niger) based dietary supplements has been investigated by HPLC-PDA, LC-ESI-MS/MS and LC-APCI-MS/MS systems. Optimization of the MS/MS parameters and LC conditions was performed using sinigrin reference standard and rapeseed certified reference material (BC190) respectively. An LC-ESI-MS/MS system was used to detect (screen) and identify the naturally occurring intact glucosinolates (GLs). The intact GLs identified were then desulfated and quantified on an HPLC-PDA system as desulfo-glucosinolates (DS-GLs). Prior to quantification, the DS-GLs were identified using an APCI-MS/MS. The HPLC-PDA method performance criteria were evaluated using glucotropaeolin potassium salt. The validated method was applied for the analysis of six dietary supplements. In total, six glucosinolates were identified and quantified in the dietary supplements; glucoraphasatin (0.2-0.48 mg/g), glucosisaustricin (0.37-0.91 mg/g), glucoraphenin (0.84-1.27 mg/g), glucoputrajivin (0.14-0.28 mg/g), glucosisymbrin (0.70-0.99 mg/g) and gluconasturtiin (0.06-0.12 mg/g). Glucoraphenin was the most abundant glucosinolate in all samples.

Determination and kinetics of enrofloxacin and ciprofloxacin in Tra catfish (Pangasianodon hypophthalmus) and giant freshwater prawn (Macrobrachium rosenbergii) using a liquid chromatography/mass spectrometry method.

Determination and kinetics of enrofloxacin and ciprofloxacin in Tra catfish (Pangasianodon hypophthalmus) and giant freshwater prawn (Macrobrachium rosenbergii) using a liquid chromatography/mass spectrometry method. J. vet. Pharmacol. Therap. 34, 142-152. The fluoroquinolones enrofloxacin (EF) and ciprofloxacin (CF) residues were investigated in the edible tissues of two important Asian aquacultured species such as Tra catfish (Pangasianodon hypophthalmus) and giant freshwater prawn (Macrobrachium rosenbergii) using a sensitive liquid chromatography-electrospray ionization-tandem mass spectrometry method. Fish and prawn were treated with medicated feed with multiple doses of EF, in field conditions. A validation study of the analytical method was realized in terms of linearity, specificity, precision (repeatability and within-laboratory reproducibility), recovery and decision limit (CCα). The time needed before the antibiotic disappears from animal tissues or reach the maximum residue limit (MRL, 100µg/kg) was assessed. The concentration values of EF detected in Tra catfish tissue were between the MRL and 2×MRL concentrations, according to the fish density, 7days following the end of the enrofloxacin treatment (20mg/kg body weight per day, for seven consecutive days). The concentration value of ER in prawn tissue was lower than the MRL and the limit of quantification (LOQ, 14µg/kg) 5 and 7days after the stop of the EF treatment (50mg/kg body weight per day, for five consecutive days), respectively. The mean detected levels of CF was much lower in comparison with that of EF, indicating that only a small part of EF is metabolized into CF (<5%) in both Tra catfish and prawn.

Modulation of CYP1A1 activity by a Ginkgo biloba extract in the human intestinal Caco-2 cells.

Ginkgo biloba is a widely consumed dietary supplement. Some dietary active compounds modulate the activity of biotransformation enzymes inside the enterocytes and more interestingly of cytochrome P-450 1A1 (CYP1A1). This enzyme is of a particular interest because of its implication in the metabolism of some exogenous pro-carcinogens or endogenous molecules. In the present work, we have used Caco-2 cells to study the effect of a standard reference material of a Ginkgo biloba extract (GBE) (10-400 µg/ml), as well as of its major individual active compounds (kaempferol, quercetin, isorhamnetin, ginkgolides and bilobalide), alone or in mixtures, at realistic intestinal concentrations, on the induction of CYP1A1 activity, in the presence or absence of benzo[a]pyrene (B[a]P) (0.1 µg/ml), a well-known CYP1A1 inducer. 3-O-rutinosides of kaempferol, quercetin and isorhamnetin were also tested. We have demonstrated a strong induction (p < 0.005) of CYP1A1 activity and a slight, but significant (p < 0.005), decrease of this activity in the presence of B[a]P by the GBE at the realistic exposure level of 100 µg/ml. The inductive effect was explained, in part, by quercetin and kaempferol after 24h exposure while unknown compounds seem to be responsible for the strong CYP1A1 induction observed after 6h exposure. The inhibitory potency of flavonols on CYP1A1 activity in presence of B[a]P was much stronger for the aglycones than for the 3-O-rutinosides, explaining the slight effect observed with the GBE, mainly composed of glycosylated flavonoids. These results indicate that GBEs may disturb intestinal CYP1A1 activity and, in turn, affect the metabolism of other compounds. The present paper thus highlights the necessity to take these side effects into account when administrating Ginkgo biloba herbal supplements.

Assessment of the chemical contamination in home-produced eggs in Belgium: general overview of the CONTEGG study.

This overview paper describes a study conducted for the Belgian Federal Public Service of Health, Food Chain Safety and Environment during 2006-2007. Home-produced eggs from Belgian private owners of hens were included in a large study aiming to determine concentration levels of various environmental contaminants. By means of the analyses of soil samples and of kitchen waste samples, obtained from the same locations, an investigation towards the possible sources of contaminants was possible. Eggs, soils, faeces and kitchen waste samples were checked for the presence of dioxins, PCBs (including dioxin-like PCBs), organochlorine pesticides, trace elements, PAHs, brominated flame retardants and mycotoxins. The study design, sampling methodology and primary conclusions of the study are given. It was found that in some cases dioxin-like compounds were present at levels that are of concern for the health of the egg consumers. Therefore, measures to limit their contamination in eggs, produced by hens of private owners, were proposed and deserve further attention.

On-farm contamination of animals with chemical contaminants.

Food products should not contain unsafe levels of chemical contaminants. However, it is not possible to monitor each and every one of the many thousands of chemicals that are used in our advanced societies. Chemical contaminants in foodstuffs of animal origin may be classified into three categories: natural contaminants (e.g. mycotoxins), environmental contaminants linked to industrialisation and/or urbanisation (e.g. dioxins and dioxin-like compounds) and authorised chemical products (e.g. residues of veterinary medical products). Chemical hazards may contaminate foodstuffs of animal origin all the way from farm to fork. Contamination may occur in any of the different production systems, and it is difficult to make comparisons between production systems (e.g. extensive versus intensive farming systems) with regard to food safety. Even when we take into account the latest analytical methods, which can detect ever-smaller quantities of residues, the relative importance of chemical contaminants seems to have declined during recent decades due to improvements in information and prevention. Nonetheless, individual incidents can never be ruled out and may have serious economic, health or social repercussions. Particular attention must be paid to chemical hazards, in order to reduce as much as possible the risks to livestock and to the consumer. Continued monitoring and periodic reassessment of risks posed by these contaminants (at the national level) are needed to detect or anticipate new problems, so that appropriate actions can be taken in the interest of public health. More attention should be paid to the production of detailed information, especially with regard to background data (e.g. the objectives of the monitoring, sampling methods, chemicals to be analysed, analytical methods, detection limits, raw data and specified units), in order to obtain a better basis for risk assessment. Such risk assessment provides control authorities with an effective tool for the exchange of information and measures to be taken to ensure food safety.

Control of the illegal administration of natural steroid hormones in the plasma of bulls and heifers.

In the context of the control of the illegal administration of natural steroid hormones in cattle husbandry, an attempt was made to establish the decision levels for sex steroid hormones in the plasma of adult cattle, taking into account the effect of the treatment. Bulls and heifers were treated with two injections, at a two week interval, of an estradiol-testosterone cocktail. Steroid hormone and biochemical precursor concentrations were measured in plasma samples by using specific radioimmunoassays, before and after the treatment. When the treatment significantly (p < 0.05) modified a hormone concentration, a decision level was established for that hormone concentration. At each decision level, a score was assigned that represented the percentage of treated animals detected when the decision limit was applied. For heifers, 17 beta-estradiol and testosterone concentrations in plasma, which increased after the treatment, are the best criteria to use to detect treated animals, with decision limits of 20 pg ml-1 and 125 pg ml-1, respectively. In the instance of bulls, both testosterone and steroid biochemical precursor concentrations decreased in the plasma after the treatment. We proposed decision limits of 1500 pg ml-1 and 28 pg ml-1 for testosterone and androstenedione concentrations, respectively, the bulls displaying concentrations below these limits being positive. We observed that the repetition of the injection increased the score of the decision limit. The scores for testosterone are 70%, 14d after the first injection and 100% 14 d after the second injection, and for androstenedione, these scores are 60 and 100%, respectively.

Purification and biochemical characterization of recombinant human placental growth hormone produced in Escherichia coli.

The hGH-V (or hGH-2) gene codes for human placental growth hormone (hPGH). Secretion of hPGH is continuous, in contrast with the pulsed secretion of pituitary growth hormone (hGH) which it progressively replaces in the maternal bloodstream. hGH-V cDNA has previously been cloned and isolated. Analysis of its nucleotide sequence has revealed a 191-residue protein, hPGH, differing from hGH at 13 positions. The calculated pI is more basic than that of the pituitary hormone. Here we have inserted hGH-V cDNA into the pIN-III-ompA3 plasmid in order to produce hPGH in its native form in Escherichia coli D1210. Expression of hGH-V cDNA in E. coli is significantly lower than that of hGH cDNA with the same expression system. The hPGH produced in E. coli was purified in quantities sufficient to allow its biochemical and immunochemical characterization. The molecular mass of the protein was determined by electrospray m.s. The determined mass, 22,320 Da, agrees well with the molecular mass calculated from the translated cDNA sequence, assuming the presence of two disulphide bridges. Having established the technique for producing hPGH with a primary structure identical to the natural, non-glycosylated, 22 kDa isoform, we can now plan the full physicochemical and pharmaceutical characterization of this new hormonal entity.

Increase in epidermal growth factor receptor and its mRNA levels by parathyroid hormone (1-34) and parathyroid hormone-related protein (1-34) during differentiation of human trophoblast cells in culture.

Human cytotrophoblasts in culture aggregate and fuse to form syncytiotrophoblasts. This process is associated with an increase in epidermal growth factor receptor (EGFR) expression [Alsat et al.: J Cell Physiol 154:122-128, 1993]. Recent studies have demonstrated the presence of parathyroid hormone-related protein (PTHrP) in the human uterus and placenta. This led us to study the effect of PTH (1-34) and PTHrP (1-34) on the expression of EGFR during this differentiation process. Both peptides induced a concentration-dependent increase in EGF binding, with a maximal effect at the physiological concentration of 1 nM. EGFR protein level assessed by cross-linking and immunoblotting and EGFR biological activity assessed by measuring its EGF-induced autophosphorylation were increased 2- and 2.5-fold, respectively, when cells were treated for 24 h with 0.1 microM PTHrP or PTH compared to control cells. This effect was time-dependent with a maximum at 3 h of treatment. This treatment also increased trophoblast cell EGFR mRNA levels, suggesting transcriptional regulation of the EGFR. To ascertain whether activation of protein kinase C (PKC) or protein kinase A (PKA) is involved in this PTH effect, we determined EGFR protein level and EGFR autophosphorylation after exposure of cells to PKA inhibitor and PKC inhibitor, alone or together with the peptide. The presence of a PKC inhibitor blocked a further increase in EGFR number by PTH, while PKA inhibitor had no effect. These results show that PTH and PTHrP increase the synthesis of EGF receptors which are strongly expressed in syncytiotrophoblasts and suggested that these peptides might be involved in human placental development.

Syncytiotrophoblastic localization of the human growth hormone variant mRNA in the placenta.

The hGH/hCS genes, clustered on chromosome 17 in the 5' to 3' order GH-N, CS-L, CS-A, GH-V and CS-B, show a high degree of sequence identity. The expression product of the GH-V gene is the placental growth hormone, which replaces pituitary GH in maternal blood throughout pregnancy. By means of mRNA competitive hybridization using 32P-labelled and unlabelled 30 bases long oligonucleotides, we first optimized specific hybridization conditions. In situ hybridization was then performed to locate the GH-V mRNA encoding placental growth hormone. The hGH-V gene appears expressed in the placental syncytiotrophoblast. Unlike the CS-A and CS-B genes (both encoding hPL) which are expressed uniformly in the syncytiotrophoblast, the GH-V mRNA is located in a few syncytiotrophoblast cells only.

Evidence for the expression of growth hormone receptors in human placenta.

Since human placenta produces a growth hormone variant, it seemed important to search for evidence of GH receptors in that organ. Evidence for the expression of the GH receptor (GHR) gene was obtained by northern blot analysis. In addition, GHR poly A+ RNA was detected in RNA from cultured trophoblastic cells, but not from placenta fibroblasts. There was a low but significant specific binding of pituitary GH-N and placental GH-V to placenta plasma membranes. Both variants apparently bound to the same receptor, which is present in the first trimester as well as in the term placenta. These results suggest that placental GH may have paracrine or autocrine functions in the placenta.

Adenohypophysis hormone gene products in 14 pituitary adenomas: analysis by immunohistochemistry and northern blotting.

We investigated 14 pituitary adenomas (10 silent adenomas; 3 prolactinomas and one GH-secreting tumor) for the presence of hormone gene transcripts (Northern blot) as well as for translation products (immunohistochemistry). The GH-secreting tumor was shown to express the genes coding for GH and PRL and to synthesize the corresponding hormones. In the cases of prolactinomas, immunohistochemical data demonstrated the synthesis of prolactin only. In addition to the PRL gene, Northern blot analysis revealed the transcription of the alpha-subunit gene in one case. Hormone genes were found to be expressed in 7 out of the 10 silent tumors, whereas no hormone synthesis was detected in any of these tissues. LH-beta mRNA was found in 3 cases, FSH-beta mRNA in 5 cases and alpha-subunit gene was shown to be expressed in one case. Surprisingly, the level of expression of the FSH-beta gene was higher than in normal tissue. This study confirms that some so called "silent" adenomas are expressing alpha- and/or beta-subunit glycoprotein hormone genes, even if no hormone is synthesized. The therapeutic action of bromocriptine described in some "silent" adenomas cases could be related to that hormone gene expression potentiality.