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In the present study, we determined circulating serum levels of human placental growth hormone (hPGH) and insulinlike growth factor binding proteins 1 and 3 (IGFBP-1 and IGFBP-3) using two-site radioimmunoassays during the gestational midtrimester of pregnancies affected by chromosomal disorders with the aim of identifying potential marker substances that might have a significant discriminative and predictive value for prenatal diagnosis of fetal chromosomal aberrations and of organ malformations such as neural-tube defect. Our results show that the maternal serum levels of hPGH were significantly elevated in pregnancies affected by chromosomal anomalies or organ malformations as compared with controls. The distribution of IGFBP-1 concentrations for all experimental groups except trisomy 21 were closely similar to the normal population. IGFBP-3 decreased slightly in pregnancies affected by Down syndrome. These findings suggest that hPGH may be useful as an additional marker in prenatal screening for Down syndrome.

In this study, we investigated the involvement of GH in rat prostate function. First, we demonstrated that specific transcripts corresponding to the GH receptor (4.5 kilobases) and to the GH-binding protein (1.2 kilobases) were expressed in the normal rat prostate, but also in all prostatic carcinoma cell lines tested (LNCaP, PC-3, MAT-Lu, MAT-LyLu, and Pif-1). Moreover, these transcripts were much more abundant in the human and rat carcinoma cells than in the normal tissue. One-year-old dwarf rats were supplemented for 7 days with saline (group DR1) or highly purified rat GH (group DR2). Northern blotting and quantitation of prostatic messenger RNAs (mRNAs) revealed that GH increases the steady state levels of transcripts coding for androgen receptor (2.4-fold), type I and II 5 alpha-reductases (2.6- and 2.2-fold), and several androgen-dependent proteins [prostatein C3 subunit (3.6-fold), probasin (11.0-fold), and R. W. B. (Royal Winnipeg Ballet) (12.5-fold)]. This suggests that GH might either potentiate the action of androgens on the prostate or act directly on this gland by a mechanism that does not depend on testicular androgens. To address this question, we supplemented hypophysectomized and castrated adult rats for 7 days with saline (group HC1), rat GH (group HC2), testosterone propionate (group HC3), or GH plus testosterone (group HC4), starting 3 days after castration. In this animal model, the abundance of C3 mRNA increased in all hormone-treated rats; the stimulation factors were 3.5 (group HC2), 25.5 (group HC3), and 9.5 (group HC4) compared to group HC1. Analysis of prostatein synthesis by Western blotting confirmed these results at the protein level. The same trend was observed for probasin and RWB mRNA levels. Probasin mRNA increased 4.5-fold in group HC2 and 12-fold in group HC3, but did not increase in group HC4 (both hormones combined); enhancement of RWB mRNA was, respectively, 5.0-, 28.0-, and 15.0-fold in groups HC2, HC3, and HC4. GH did not affect the abundance of androgen receptor mRNA. As described previously, the level of this mRNA dropped significantly in group HC3. GH alone did not significantly alter the level of either 5 alpha-reductase mRNA, whereas testosterone, alone or with GH, produced a 2-fold increase in type II 5 alpha-reductase mRNA (groups HC3 and HC4). Type I isoenzyme mRNA reached 1.6 times the control level (group HC1) in groups HC3 and HC4.(ABSTRACT TRUNCATED AT 400 WORDS)

We have studied dose-dependent effects of highly purified human PRL (39 IU/mg) on the testis of immature (22-day-old) hypophysectomized rats daily supplemented for 7 days with 2, 10, or 30 micrograms hormone. Dose-dependent stimulation was observed for all parameters: testis weight (1.6- and 2-fold above control for 10 and 30 micrograms PRL), basal and hCG-stimulated testosterone (14- and 21-fold), intratesticular testosterone (7- and 21-fold) and estradiol (1.2- and 1.5-fold), LH receptor concentration (1.8- and 2.5-fold), in vitro pregnenolone production by cholesterol side-chain cleavage enzyme (3-, 5- and 7-fold), and aromatase activity (2.1- and 2.4-fold). The number of Leydig cells exhibiting immunoreactivity toward anti-P450scc, anti-P450(17 alpha), and anti-3 beta-hydroxysteroid dehydrogenase antibodies also underwent a dose-dependent increase (under conditions revealing many immunopositive cells in hypox control animals). The respective increases were 8- to 14-fold for anti-P450scc and P450(17 alpha) and 1.5- to 2-fold for anti-3 beta-hydroxysteroid dehydrogenase. The number of germ cells and the percentage of tubular sections containing late pachytene spermatocytes as most advanced stages of spermatogenesis were subject to similar dose-dependent effects. These results suggest that during puberty PRL stimulates testicular function by promoting multiplication and differentiation of Leydig cells (acting at various steps of steroidogenesis and on tissue responsiveness to LH) and germ cells.

Familial (hereditary) haemochromatosis (HH) is an iron storage disorder characterized by an increased intestinal absorption of iron and its accumulation in numerous tissues. The disease generates an iron overload with tissue damages also seen in haematologic disturbances (with dyserythropoiesis and haemolysis) and hepatic disorders. Besides typical mutations linked to HH (C282 Y and H63D, HFE locus), three other mutations have been identified and more have to be defined. A complete genetic testing is important to assess the risk of morbidity. Indeed, the clinical picture of HH is dependent upon the specific mutations as well as the individual context (sex, environment, associated hepatic and/or haematologic disorders). Porphyria cutanea tarda (PCT) has for long been found significantly associated with HH. It is now considered that hepatic iron overload related to the combination of heterogeneous genetic traits and environmental factors, including alcoholism and viral hepatitis, precipitates the expression of PCT through the inhibition of uroporphyrinogen decarboxylase (Uro.D).

A patient, born in 1961, was first examined for autoimmune hyperthyroïd at the age of 25. Then, several episodes of serious dehydration and alteration of general condition were followed by a severe encephalopathy with a symptomatology of stroke-like episodes, and during a hospitalization, a volvulus of the colon. Epilepsy developed also. A MELAS syndrome was diagnosed when she was 32 years old, based on her clinical metabolic history of the presence of Ragged-Red-Fibers at muscular biopsy and of the genetical analysis. At the age of 37, after a new episode of encephalopathy, the patient fell into a rapidly evolving coma and deces.

The diagnosis of Cushing's syndrome remains a challenge in clinical endocrinology. Cushing's syndromes are usually classified as dependent or independent from ACTH. In the first class are Cushing's disease, the ectopic corticotropin syndrome and the rare ectopic CRH syndrome. These ACTH-dependent Cushing's syndromes usually present diffusely hyperplastic adrenal glands. In the second class, are cortisol producing unilateral adrenocortical adenomas or carcinomas. New entities have recently emerged as bilateral adrenal hyperplasia not dependent from ACTH; their etiopathogenies are heterogeneous with illicit expressions at the adrenal level of functional receptors to various ligands: GIP, catecholamines, lutropin... The knowledge of such entities has to be taken into consideration in the diagnostic and management of ACTH independent Cushing syndromes.

The respective contributions of pituitary and placental GH to circulating IGF-I in pregnant women have not been well established. We measured the serum concentrations of placental growth hormone (PGH) and IGF-I in a woman with pit-1 deficiency before, during and after pregnancy, resulting in the birth of a healthy child (not pit-1 deficient). Both PGH and IGF-I concentrations were below the assay detection limit before and after pregnancy. During pregnancy, PGH and IGF-I levels increased steadily; the concentrations of PGH and IGF-I in late pregnancy were comparable with levels previously measured in normal pregnancies. PGH and IGF-I concentrations were strongly correlated throughout pregnancy (r = 0.90; P = 0.002). PGH was undetectable in cord serum, whilst the IGF-I concentration was within the normal range. The findings of this case study corroborate the notion that PGH is the prime regulator of maternal serum IGF-I during pregnancy.

We have cloned a novel complementary DNA whose expression was decreased in rat Sertoli cell cultures after treatment with FSH. This complementary DNA encodes a protein of 570 amino acids and shares 92% homology with the human MAGE-D protein. In contrast to other MAGE genes (A, B, or C), we have shown that MAGE-D expression was ubiquitous in healthy rat tissues. In the seminiferous tubules, the MAGE-D was expressed in Sertoli cells but not in germ cells as demonstrated by RT-PCR and in situ hybridization, whereas for the other MAGE genes, expression has been shown to be restricted to germ cells. Interestingly, MAGE-D was also detected for the first time in the female gonad by Northern blotting. In MLTC-1 cells (mouse Leydig tumor cell line-1), LH and PRL stimulated MAGE-D expression. Using hypophysectomized rats, it was confirmed that FSH decreased MAGE-D expression, whereas LH and PRL increased MAGE-D messenger RNA level in the whole testis most probably through a direct action on Leydig cells. As MAGE-D is present in both the seminiferous compartment and interstitium and hormonally regulated in each, it is possible that it has specific functions in each compartment during the development and the maintenance of the testis.

We have identified a novel complementary DNA (cDNA) corresponding to a gene overexpressed in the rat ventral prostate after castration. This cDNA displays 89.4% identity with 453 bp of a mouse EST and 81.5% identity with 157 bp of a human EST and was named PARM-1 for prostatic androgen-repressed message-1. The complete cDNA is 1187 bp long and codes for a protein of 298 amino acids that contains four potential glycosylation sites and three half cystinyl residues. The PARM-1 gene was found to be expressed at quite low levels in most rat tissues including those of the urogenital tract. The kinetic of induction of PARM-1 gene in the prostate was highly correlated to the development of apoptosis in the whole organ. Supplementation of castrated animals with androgens reversed both the process of apoptosis and the overexpression of PARM-1 gene. Supplementation with estrogens did not result in an increase in the PARM-1 messenger RNA levels when compared with the castration alone. However, the treatment resulted in a more rapid return to intact levels in the castrated plus estrogen group. When apoptosis of testis and prostate was induced in vivo by hypophysectomy, it was found that PARM-1 was only overexpressed in the prostate. Therefore, PARM-1 seems to be regulated by androgens only in the prostate. Using in situ hybridization and immunohistological techniques, we have shown that PARM-1 gene product is found exclusively in the epithelial cells of involuting prostate. Analysis by flow cytometry of MAT LyLu epithelial cells transiently expressing PARM-1 protein did not allow us to demonstrate a direct effect of PARM-1 gene overexpression on the programmed death of the transfected cells. Treatment of MAT LyLu cells by transforming growth factor-beta induced apoptosis but had no effect on PARM-1 production. However PARM-1 protein has been detected by Western blotting in various cell lines such as MAT LyLu, MAT Lu, and PIF, which are androgen independent. This would suggest that PARM-1 gene product would be a marker for acquired androgen-independence of these tumor cells.

This work present the results of the use of 131I, as first line and only, antithyroid treatment, applied to 120 patients suffering from autoimmune hyperthyroidism, compared to 64 patients with toxic goiter and adenoma. The diagnostic weight of the determination of the various antithyroid antibodies was assessed in the identification of autoimmune hyperthyroidism, for which TRAb and TPO Ab are the most significant. The evolution after treatment of a preexisting ophthalmopathy (i.e. Graves'disease) was never found to be alarming and in most cases regularly improved with time. A single case (out of 72) of autoimmune hyperthyroidism with no preexisting sign developed a severe ophthalmopathy after treatment, which was well controlled thereafter. Mean cumulated 131I doses to control hyperthyroidism were 22.8 mCi for toxic goiter, 21.6 mCi for adenoma, 14.1 mCi for autoimmune hyperthyroidism and 16.7 mCi for Graves'disease. Post 131I hypothyroidism was found in 28.2% of toxic goiters, 12.1% of adenomas, 66% of autoimmune hyperthyroidisms and 70% of Graves'disease. No relapse was observed after treatment. A surgical indication was proposed for cases requiring more than two 131I doses to control hyperthyroidism.

Although essential, androgens alone are not sufficient to induce normal growth and functionality of the prostate. Nonandrogenic hormones must also be involved in the proliferation of the prostate cancer cells which do not respond to antiandrogenic therapy and which thus become androgen-independent. Prolactin, but also growth hormone and luteinizing hormone, are potentially able to act on both normal and abnormal prostatic cells.In this review we summarize data from the literature concerning the physiological and pathological implications of prolactin, growth hormone, and luteinizing hormone on the prostate.In rodent prostates, prolactin and growth hormone can induce a variety of effects independently of androgens (e.g., transactivation of certain genes, or synthesis of the major secretion products). Moreover, hyperprolactinemia is responsible for inflammation and dysplasia of the gland, while growth hormone promotes the development of prostate tumors in vivo in the mouse and rat. Growth hormone acts on the gland directly, through prostatic growth hormone receptors, and/or indirectly via the stimulation of insulin-like growth factor-I (IGF-I) synthesis in the liver. Luteinizing hormone receptor is expressed in rat and human prostates. Luteinizing hormone increases the amount of various transcripts in the rat prostate through an androgen-independent pathway.Prolactin, growth hormone, and luteinizing hormone, alone or synergistically with androgens, play physiologically significant roles in the normal prostate. The involvement of these hormones in the development of benign prostatic hyperplasia and prostatic carcinoma is an issue that needs to be addressed.

The various types of diabetes mellitus are now more precisely classified according to their etiopathogenies. Therefore, type 1 (autoimmune or not), type 2, gestational diabetes, as well as MODY (Maturity Onset-Diabetes of the Young) and secondary diabetes are considered as separate entities. Any type of diabetes brings its set of complications. Biological chemistry is directed to define the etiopathogenic characteristics of the various types of diabetes and to assess the severity of metabolic and functional disturbances. Diagnostic criteria in term of glycemia have also been reviewed to asses diabetes and glucose intolerance.

The objective was to elicit whether hormonal responses to and subjective symptoms of hypoglycemia are modified during pregnancy in patients with insulin-dependent diabetes mellitus.In ten type I diabetic women, hyperinsulinemic hypoglycemic clamps, with target arterial blood glucose content of 2.2 mmol/l, were performed during the third trimester of pregnancy and 5-13 months after delivery. The levels of arterial glucose and venous catecholamines, pituitary growth hormone, cortisol, glucagon, dehydroepiandrosterone and free insulin were assessed repeatedly. Subjective symptoms of hypoglycemia were recorded on a visual analog scale. The paired t-test and Wilcoxon signed rank test were used for statistical comparisons.Adrenaline and dehydroepiandrosterone increased during hypoglycemia on both occasions, but dehydroepiandrosterone disclosed a significantly different pattern of response during pregnancy (p=0.016). The cortisol increase was augmented during pregnancy (420+/-50 vs. 310+/-30 nmol/l non-pregnant, p=0.039), while the increase in pituitary growth hormone (hGH) was diminished (5.4+/-1.2 vs. 27.9+/-5.4 microg/l non-pregnant, p=0.001), but nevertheless the increase during pregnancy was significant (p=0.002). Three of the eight subjective symptoms of hypoglycemia recorded were less prominent during pregnancy, namely 'inability to concentrate' (p=0.03), 'headache' (p=0.01) and 'pounding heart' (p=0.03).Albeit some subjective symptoms were diminished during pregnancy, the study gives no evidence that, in diabetic patients, pregnancy per se impairs the counterregulatory response to hypoglycemia, with the exception of growth hormone. However, despite the suppressed basal growth hormone secretion in late pregnancy, the study disclosed a secretory response of this hormone at hypoglycemia.

In pregnancy, the human placenta GH acts as a growth-promoting hormone and appears to be the main stimulator of insulin-like growth factor I (IGF-I) secretion. In a woman with a TSH-secreting macroadenoma, successful treatment with the somatostatin analog octreotide was conducted during the first month and the second half of pregnancy without side-effects on placental and fetal development. As observed in normal pregnancy, both serum placental GH and IGF-I levels increased throughout pregnancy and dropped sharply after delivery. In placental membranes from both treated and healthy untreated patients, we demonstrated the presence of high affinity binding sites for somatostatin-14 (Kd, 4.6 and 5.3 nmol/L; binding capacity, 1.53 and 1.35 pmol/mg protein, respectively). These receptors displayed low affinity for octreotide (IC50, 1.2-2 mumol/L), suggesting the presence of SST1 and/or SST4 receptors. We found that messenger ribonucleic acids of these two subtypes were expressed in both human placental tissue and purified human cytotrophoblast cells. Finally, the SST1-selective analog, des-AA1,2,5[D-Trp8,IAmp9]S-14 had low affinity for placental somatostatin receptors. These results argue in favor of the presence of the SST4 subtype in human placenta. At the doses administered, octreotide did not bind to placental somatostatin receptors. Our results may explain the absence of changes in both human placental GH and IGF-I concentrations that we observed during octreotide treatment.

Ein ischamischer Insult ist definiert durch eine akute fokale Durchblutungsstorung des Gehirns, die sich in der Regel durch ein akut auftretendes umschriebenes neurologisches Defizit, welches fluchtig oder dauerhaft sein kann, bemerkbar macht. Ein ischamischer Infarkt ist Ausdruck einer bleibenden strukturellen ischamischen Hirnschadigung.

The objective of this study was to determine the effects of pituitary hormones (luteinizing hormone [LH], follicle-stimulating hormone [FSH], growth hormone [GH], and prolactin [PRL]) on interstitial cell proliferation and differentiation in the testis of immature hypophysectomized rats. Macrophages, Leydig cells, precursor mesenchymal cells, endothelial lymphatic cells, and myoid cells were studied. Our experimental approach was aimed at determining whether changes in a cellular subpopulation observed after pituitary hormone treatments were the result of division of existing cells in the population, of differentiation of interstitial precursor cells, or both. In this context, it must be stressed that our data reflected the effects of hormones to prevent the decline of cells due to hypophysectomy rather than their recovery. Macrophage proliferation was taken into account because macrophages closely resemble Leydig cells and are known to proliferate after hormonal treatment. A double-labeling procedure (acid phosphatase and anti-bromodeoxyuridine [anti-BUdR]) revealed that LH, FSH, and PRL increased the number of testicular macrophages 105-, 104-, and 103-fold, respectively, in hypophysectomized rats compared to hypophysectomized control animals. BUdR incorporation in testicular macrophages was greater after PRL treatment than after LH and FSH supplementation. In contrast, we were unable to demonstrate any effect of rat GH on the macrophage population. Light microscopic analysis of plastic embedded sections of treated rat testis revealed that LH increased the numbers of Leydig, precursor mesenchymal, and myoid cells 6-, 4-, and 1.3-fold, respectively. LH also stimulated BUdR incorporation into all interstitial cell types. PRL administration increased both the number of Leydig and precursor mesenchymal cells (each 3-fold) but decreased the number of endothelial lymphatic cells (1.5-fold) when compared to the control animals. In contrast, FSH did not increase the number and proliferation of Leydig cells but exerted a slight proliferative effect on the other interstitial cell populations. In GH-treated rats, the number of precursor mesenchymal cells increased two fold above the control rats. GH also exerted slight proliferative effects on both precursor mesenchymal and myoid cells. Immunohistochemical studies of steroidogenic enzymes in the testicular interstitium of treated rats demonstrated the presence of steroidogenic enzymes, not only in Leydig and precursor mesenchymal cells, but also in some (1%-2%) endothelial lymphatic cells and myoid cells. This may indicate that both of these cell types are also constitutively equipped to perform steroidogenesis or that they are precursor cells undergoing differentiation. Taken together, changes in the number of Leydig cells in our animal model appeared more likely to be dependent on the transformation of precursor cells than on division of preexisting mature Leydig cells.

During pregnancy, the human placenta secretes a variant of pituitary growth hormone (hGH-V) differing in sequence from the pituitary growth hormone (hGH-N) by 13 amino acids substitutions. HGH-V replaces hGH-N in the maternal bloodstream during the second half of pregnancy. HGH-V is produced in two, possibly three, isoforms: a 22 kDa form, a glycosylated 25 kDa form, and a probable 26 kDa form resulting from alternative splicing of the mRNA. Here we have studied the somatogenic and lactogenic activity of recombinant 22 kDa hGH-V isoform produced in Escherichia coli and compared it with the 22 kDa form of hGH-N. The two variants exert a similar somatogenic effect on rabbit and rat cells, but differ as regards their lactogenic activity in the rat.

In addition to androgens that are essential for maintenance of prostate growth and function, nonandrogenic hormones must also be considered to explain some regulatory events occurring in the prostate. The detection of LH/CG receptor (LH/CG-R) gene expression in some nongonadal tissues, has led us to consider LH as a potential regulatory factor of prostatic development and function. In this study, we have demonstrated by RT-nested polymerase chain reaction and Northern blotting that the rat prostate contains the same LH/CG-R transcript as the gonads. Western immunoblotting, ligand blotting, and binding analysis have shown that rat prostate also contains a 93-kDa receptor protein that is able to bind [125I]hCG specifically and with a high affinity (6.0 x 10(9) M-1). Our results also indicated that the concentration of binding sites was lower in the prostate than in the gonads. LH/CG-R sites of expression have been localized in the prostate by immunohistochemistry: specific staining was observed in all the epithelial cells of the gland, but the ventral lobes are much more immunoreactive than the lateral and dorsal lobes. Finally, the ability of this prostatic LH/CG-R to induce a physiological response was evaluated in an explant culture system. A time course experiment was carried out, and we observed a significant dose-dependent stimulation of cAMP production after 3 h of treatment: 3.0 +/- 0.4, 4.2 +/- 0.4, and 5.0 +/- 0.5 pmol/ml of cAMP for 0, 100, and 500 ng/ml of LH, respectively. In conclusion, LH is able to act directly on the prostatic gland through specific receptors that are structurally and functionally very similar to those expressed in the gonads and are mainly localized in the ventral lobe of the organ. These data suggest that LH plays a significant physiological role in the prostate.

Consideration of the abnormal regulation of fetal growth leading to intrauterine growth retardation must take account of the fundamental differences between the regulation of growth before and after birth. The significance of endocrine regulators of growth differs greatly in utero. During the first trimester of pregnancy, embryonic growth might be controlled at the level of the individual organs by nutrient supply and by locally active growth factors. Later, fetal growth depends essentially upon materno-placental cooperation in delivering nutrients to the fetus. Therefore the major role of hormones in fetal growth is to mediate utilization of available substrate. Fetal growth seems to be regulated by fetal insulin, IGF-1 and certainly IGF-2, while growth hormone has only a secondary role to play. In late gestation, placental size and fetal growth rate are well correlated, pointing to a key role of the placenta in the regulation of fetal growth. It is therefore of importance to understand the molecular mechanisms involved in regulating placental development and endocrine functions. TGF alpha and EGF might play a major role as suggested by the modulation of their receptors with placental development, and by the specific alterations of epidermal growth factor receptors in intrauterine growth retardation. In addition, human placenta secretes specifically placental growth hormone. The concentration of placental growth hormone is significantly decreased in sera of pregnant women bearing a fetus with intrauterine growth retardation.

A brief review of the cancers derived from the follicular thyroid epithelium and parafollicular cells (medullar cancer) is made. The contribution of clinical biology to the diagnosis has little significance in the case of thyroid cancer. On the other hand, the quantitative analysis of thyroglobulin is very important for the detection of recurrence in the course of the follow-up of a patient who has been treated by complete thyroidectomy. The biological markers of medullar cancer are most specific. The observation of pathological secretion of thyrocalcitonin is determining for the diagnosis of medullary carcinoma. It must lead to a parathyroid and medulloadrenal exploration as well as to systematic investigation of family history. These steps are essential for the diagnosis of the syndromes of multiple endocrine neoplasias types 2a and 2b.

In many instances amiodarone-induced hyperthyroidism has been reported as mild, thyroid functions returning to normal after discontinuation of the drug. Nevertheless, life-threatening amiodarone-induced thyrotoxicosis has also been described. Conventional treatments such as with antithyroid drugs (Thionamide) and corticosteroids are essentially ineffective or fail to stop the dramatic course of the thyroid crisis. This limited efficacy of medical therapy, particularly in patients with previously--neglected or unknown--thyroid disease, prompted us to intervene surgically. We report a series of six patients who underwent total or nearly total thyroidectomy as first line therapy for four of them. Surgery resulted in rapid resolution of thyrotoxicosis with an uneventful postoperative course. This approach has the advantage of immediate and safe efficacy, low risk of relapse and finally, appears to be the only antithyroid treatment that permits continued therapy with amiodarone.

Insulin-like growth factor I (IGF-I) has been purified from defatted cow colostrum. The purification procedure involved cation exchange chromatography on CM Affigel blue, acid gel-filtration and two HPLCs. This purification process constitutes a significant improvement, in terms of yield and rapidity, over the previously reported procedures. Purified IGF-I was found to be 95% pure by N-terminal amino acid sequence analysis. After isolation of peptides from a digest of IGF-I obtained using staphylococcus aureus protease, the complete sequence was established and found to be identical to that of bovine or human plasma IGF-I.

In this study we have compared the effects of the homologous rat growth hormone with its human and bovine counterparts on the testicular development of immature hypophysectomized rats, after 7 days of treatment. Each hormone increased (12- to 13-fold) the body weight gain and IGF-I blood level (12- to 16-fold). Human GH increased testis weight (1.3-fold) the pregnenolone production (4-fold), the LH dependent steroidogenesis (7-fold), the aromatase activity (5-fold), the concentration of LH receptors (1.6-fold) and spermatogenesis. Bovine and rat growth hormones had marked effects only on pregnenolone production (2- to 3-fold). Incorporation of 5-bromodeoxyuridine in testicular cells after hGH treatment was positive (0.47 per 100 cells), an effect which was 2-fold higher than those produced by bovine and rat GHs (0.25 per 100 cells). These results have led us to conclude that the in vivo action of rat growth hormone on rat testis is comparable to bovine GH and thus might be considered as purely somatogenic. In addition to the known effects on growth promotion, rat GH affected the rate limiting step of steroidogenesis and the Leydig cell multiplication.

A GH variant of placental origin, placental GH, has recently been shown to replace pituitary GH in maternal serum during pregnancy. Besides, the GH variant (GH-V) gene has been demonstrated to be expressed in the placenta. The similarities between their known properties strongly suggest that the placental GH and the GH-V protein are the same molecular species. Here we provide final evidence that this is indeed the case by sequence analysis of both the 22K and 25K forms. Furthermore, the 25K form is shown to be glycosylated, while the 22K form is not. Both size variants of placental GH are, thus, likely to reflect the partial glycosylation of a unique pep-tidic chain.

SummaryThe determination of serum thyrotropin (TSH) by radio-immunoassay is now part of the usual investigations performed for patients suffering from thyroid disorders.We describe here an original radioimmunoassay of h.TSH using an antiserum raised against porcine thyrotropin and a highly purified h.TSH prepared in our laboratory as tracer and standard.Immunological analysis of this system demonstrates its complete hormonal specificity. Its sensitivity is better than that obtained with most of the techniques described by others, and allows the detection of 4. 10-5 pmole of the hormone.Clinical material resulting from a two year period of use is presented and the usefulness of the method is emphasized for the study of thyroid and hypothalamopituitary pathology as well as in the adjustment of suppletive or antithyroid treatments.

In an attempt to localize topographic domains specific to native human chorionic gonadotropin (hCG), we studied the discontinuous antigenic regions recognized by a monoclonal anti-hCG antibody designated as C8 which binds only to hCG and does not cross-react with either the free hCG-alpha and hCG-beta subunits or other glycoprotein hormones. Using two-site monoclonal immunoradiometric assays (M-IRMAs), we found that C8 antibody and an anti-peptide antibody (FB12) directed to residues 110-116 of hCG-beta did not bind simultaneously to hCG. This observation suggested that C8 binds to residues of hCG-beta included either in the antibody-binding region of FB12 or in close proximity to amino acids 110-116. To further delineate the regions of hCG-beta recognized by C8, we carried out hapten inhibition experiments with synthetic peptides corresponding to various regions of hCG-beta. The peptide corresponding to residues 109-122 and subpeptides (111-122 or 112-122) inhibited the binding of 125I-hCG to C8, whereas weak inhibition was observed with subpeptide 113-122. By studying the binding of C8 to the 1-112 disulfide-bonded part of hCG-beta (hCG-beta core) recombined with hCG-alpha, we were able to confirm that C8 binds to a region including or near to Asp112. M-IRMAs showed that C8 does not bind to the recombinant molecule lacking residues 113-145 of hCG. Taken together, these results indicate that a limited number of residues located on hCG-beta near to Asp112, and most likely the sequence Asp111-Asp112-Pro113, are included in the discontinuous antigenic region recognized by C8. We then attempted to localize residues of hCG-alpha that constitute another part of the determinant which bound to C8. Six synthetic peptides corresponding to various regions of hCG-alpha did not inhibit binding of 125I-hCG to C8. In contrast, M-IRMAs demonstrated that C8 is capable of binding recombinant products composed of the hCG-beta subunit and the alpha subunits from human, equine, and porcine species. These results indicate that C8 recognizes a region of the alpha subunit highly conserved in these three species. Finally, we determined that the discontinuous regions recognized by C8 are partially accessible on the CG/LH-receptor complex.

Serum profiles of growth hormone (GH), hPL and hCG were recorded in 5 pregnant women during termination of pregnancy at cesarean section and legal abortion. Two monoclonal antibodies directed against different epitopes on the GH molecule were used to distinguish the pituitary 22K-GH from the placental GH variant. After placental removal at cesarean section, a rapid fall in GH as well as in hPL and hCG concentrations was recorded. On the other hand GH levels were unaffected when early pregnancy was terminated by legal abortion. The importance of an intact placental function for the maintenance of high GH levels lends further support to the concept that a placental GH variant is produced during late pregnancy.

This work was undertaken to study the heterogeneity of GH in serum and placental and pituitary extracts and to study GH physiology in pregnant women. Two distinct monoclonal antihuman GH (anti-hGH) antibodies (MAb) coded 5B4 and K24 were selected for their high binding affinity and specificity. The 5B4 MAb recognized the epitope comprising the NH2-terminal end of hGH, and the K24 MAb recognized an internal epitope. Both MAbs were used in RIAs to measure serum GH concentrations in various circumstances, including pregnancy. The two RIAs yielded slightly different serum GH results in normal men and nonpregnant women, but the overall correlation between the data was excellent. Since the RIAs were not affected by human placental lactogen, the evolution of serum GH in pregnant women could be studied. In such women, serum GH levels progressively declined to undetectable levels during the second half of pregnancy, while a pregnancy-associated serum GH-like antigen [tentatively called human placental growth hormone (PGH)] appeared in the circulation at midpregnancy and increased thereafter up to term. PGH contained the NH2-terminal epitope of pituitary GH, but lacked the internal one. Consequently, it reacted selectively with the 5B4 MAb only. After delivery, PGH disappeared from maternal serum within 1 h. Amniotic fluid contained low GH concentrations; cord serum contained high GH levels, but no PGH. Thus, PGH appears to be secreted selectively into the maternal compartment. PGH was purified from term placenta extracts. According to its chromatographic behavior, it appears more basic than pituitary 22K and 20K GHs. Size dimorphism was demonstrated; PGH was composed of two entities of 22K and 25K, respectively. Pure PGH, obtained in small quantities by preparative electrophoresis, was found to bind to hepatic GH receptor with an apparent high potency compared to that of pituitary GH, PGH, thus, should act in vivo as a GH agonist sharing most of its biological properties. These results lead to the conclusion that PGH is likely to replace the pituitary hormone in governing maternal metabolism during the second half of pregnancy.

The effects of pure FSH and/or LH preparations on the number of Leydig cells and their function in immature hypophysectomized rats have been investigated. As a result of hypophysectomy at the age of 17–18 days, the number of recognizable Leydig cells per testis decreased, as did the steroidogenic capacity in vivo and in vitro. Treatment with 64 μg FSH on both 22 and 23 days of age, did not affect the number of recognizable Leydig cells. In contrast, two injections of LH (10 μg) caused a sixfold increase in the number of Leydig cells, but had a negative effect on spermatogenesis. These stimulatory and inhibitory effects of LH diminished when FSH was added. Treatment with FSH for 7 days caused a twofold increase in the number of Leydig cells when compared with hypophysectomized controls. 3β-Hydroxysteroid dehydrogenase (3β-HSD) and esterase activity in Leydig cells also increased under the influence of FSH. The pregnenolone production per Leydig cell in the presence of 5-cholesten-3β,22(R)-diol (22R-hydroxycholesterol) as substrate showed a sevenfold increase. Plasma testosterone levels 2 h after injection of human chorionic gonadotrophin in intact rats and hypophysectomized FSH-treated rats were the same. Following LH treatment for 7 days, the number of Leydig cells proved to be 11 times higher, and 3β-HSD and esterase activity were not different from intact controls. The testicular pregnenolone production was four- to fivefold higher when compared with untreated hypophysectomized rats. However, pregnenolone production per Leydig cell in LH-treated rats was only slightly different from the hypophysectomized controls. In conclusion, FSH treatment caused an increase in the number and steroidogenic activity of Leydig cells, and LH had a major effect on the number of Leydig cells, but did not stimulate the steroidogenic capacity. Journal of Endocrinology (1989) 120, 97–106

Changes in the concentrations of LH and FSH testicular receptors have been studied in the pig, from neonatal to adult life, and correlated with blood LH, FSH and testosterone concentrations. Quantification of gonadotrophin receptors was performed in equilibrium binding studies, using homologous systems. The presence of high-affinity binding sites for LH and FSH (association constant (Ka): LH ∼ 20 litres/nmol; FSH ∼ 10 litres/nmol) was demonstrated in the testes of all animals studied. The apparent affinity of LH and FSH receptors did not change significantly with age. During the first weeks of life, there was a transient rise in LH receptor content, reaching a maximum of 8·7 ± 2·2 (mean ± s.e.m.) pmol/g testis at 24 days of age. This was correlated with a peak in testosterone secretion and reflects the second wave of interstitial cell proliferation in the pig. A second increase in the number of LH receptors occurred after 12 weeks of age and corresponds to pubertal maturation and final differentiation of adult Leydig cells. During this period, circulating concentrations of testosterone markedly increased without any significant variation in LH blood levels, suggesting a change in testicular sensitivity to LH in the maturing pig. A continuous increase in FSH receptor content was observed from the neonatal to the adult pig. This increase occurred in two phases. During the first 2 months of life, the increase in the number of FSH receptors exceeded that of testis growth rate and resulted in an increase in FSH receptor concentrations which reached a peak at 12·1 ± 1·8 pmol/g testis, at week 9. After this time the increase in the amount of FSH receptors was slower than that of testis weight gain, leading to a decrease in receptor concentration. Since changes in the amounts of LH and FSH receptor concentrations were not correlated with variations in gonadotrophin concentrations in the blood, an inductive role for FSH on LH receptors, as has been shown in the rat, could thus not be demonstrated in the pig. J. Endocr. (1986) 111, 301–308

The properties of follitropin receptors in immature porcine testis were determined using highly purified porcine follitropin. 1. The characteristics of follitropin binding to a subcellular fraction rich in plasma membranes were studied using a 125I-labelled follitropin with high specific activity (75-100 Ci/g) and high binding activity. The binding is dependent on time, temperature and pH. It is specific to follitropin as demonstrated by the very low binding activity of the follitropin alpha and beta subunits and of the other glycoprotein hormones. Scatchard analysis of binding data indicated an equilibrium association constant of 2 x 10(10) M-1 and a concentration of high affinity binding sites of 500 fmol/mg membrane proteins. 2. A sensitive radio-ligand receptor assay was developed. Fifty percent inhibition of binding was obtained with as little as 2 ng of porcine follitropin. Ovine and bovine follitropins and pregnant mare serum gonadotropin gave binding inhibition curves parallel to that given by porcine follitropin. With equine and human follitropin, significantly different slopes were recorded. 3. Kinetics of dissociation of labelled follitropin from its testis receptors showed the presence of at least two compartments with fast and slow dissociation rate constants. The ratio between the sizes of the slow and fast compartments appeared dependent upon preincubation time. 4. A temporal correlation was observed between binding of follitropin to testis receptors and activation of membrane bound adenylate cyclase.

The 20K variant of native (22K) hGH is a full agonist for the growth promoting and lactogenic properties of the hormone in vivo but has been reported to have weak or absent insulin-like properties. To explore if these differences may be explained at the receptor level, we compared the ability of 22K and 20K hGH to inhibit the binding of 125I-22K hGH to receptors in isolated rat adipocytes, a target for the insulin-like effects of the hormone and in IM-9 cultured human lymphocytes, more specific for growth effects. Our data show that while 20K hGH is a potent agonist of native 22K hGH in the IM-9 lymphocyte assay, its potency in the rat adipocyte binding assay is only 3%, even when both cells are incubated together in identical conditions. Thus, the receptors for hGH appear to be different on various target cells, explaining why the 20K variant has different relative biological potencies at different sites of action.

The gonadotrophin preparations that have been used previously to study different aspects of testis function are of limited purity. We have, therefore, purified existing gonadotrophin preparations further. The demonstration of their purity and biological activity are reported as well as their suitability for in-vivo use. After separation of the subunits of the intact hormones by high-performance liquid chromatography followed by analytical sodium dodecylsulphatepolyacrylamide gel electrolphoresis, no contaminating proteins could be detected in either the FSH or LH preparation. After immunoadsorption, contamination by other pituitary hormones (TSH, prolactin, FSH, LH and GH) was found to be less than 0·002% by weight for all the hormones tested. The biological activity of porcine FSH (pFSH) measured in a Steelman–Pohley assay was 150–170 times more potent than the NIH-FSH-P1 reference preparation. The biopotency of human LH (hLH) in the ovarian ascorbic acid depletion test was measured and appeared to be 8100–8300 IU/mg against the 68/40 International Standard. The dose-dependent effects of pFSH and hLH on testis weight and the number of FSH and LH receptors were measured in immature (22-day-old) hypophysectomized rats treated for 7 consecutive days. Treatment with FSH induced a dose-dependent increase in testis weight (threefold) when compared with the control. The concentration and total number of LH receptors were increased (two- to sixfold) in a dose-dependent manner. The number of FSH receptors per testis increased while the number of FSH receptors per mg of protein remained unchanged. Administration of human LH to immature hypophysectomized rats had no effect on either LH or FSH receptors, regardless of the dose administered. The plasma concentration of testosterone measured after seven injections of FSH remained at the level of (0·31 nmol/l) found in saline-injected control rats. After 7 days of treatment with hLH, the plasma concentration of testosterone had increased in a dose-dependent manner (three- to tenfold). These results indicate that the gonadotrophin preparations are both highly purified and retained full biological activity and, therefore, are ideal for in-vivo physiological studies. Journal of Endocrinology (1989) 120, 89–96

1. The partial purification and immunological characterization of the human chorionic thyrotropin (HCT) was described. The behavior of the chorionic factor during the steps of purification was similar to that of pituitary thyrotropin (TSH) although its chromatographic and electrophoretic properties suggested that HCT was less acidic than the human pituitary hormone. The specific activity of the most purified fraction was 3500 times that of the starting extract. 2. Double diffusion experiments showed the extent of immunological cross-reactivity of HCT with human pituitary thyrotropin to be low, in confirmation of earlier results, but a close immunological relationship between the chorionic factor, bovine and porcine TSH was found when both were reacted against highly specific antibovine or antiporcine TSH serum. This rather surprising difference in the degree of cross-reactivity between HCT with human compared to that with porcine and bovine TSH was confirmed by experiments on the neutralization o...

A contercurrent distribution system has been devised for bovine thyrotropin which permits easy removal of several impurities, including one of the peptide chains of luteinizing hormone. The remaining chain of luteinizing hormone, which has a number of properties similar to those of thyrotropin, can then be removed by gel filtration. The thyrotropin is biologically active after the countercurrent distribution and no evidence has been found for the separation of subunits as occurs with luteinizing hormone. The active thyrotropin contains more methionine, tyrosine, and lysine than previously reported. Electrophoresis of the reduced and S-carboxamidomethylated derivatives of thyrotropin and luteinizing hormone in urea-containing gels reveals polymorphism beyond that exhibited by the native hormones. In the case of thyrotropin, the added polymorphism does not seem to represent dissociation into subunits. Polymorphism of native luteinizing hormone obtained as a by-product of the preparation of thyrotropin has also been found. Because the luteinizing hormone which is most difficult to separate from thyrotropin by chromatography cannot be distinguished from thyrotropin by conventional gel electrophoresis, new electrophoretic criteria have been developed to establish when thyrotropin preparations are free of luteinizing hormone or either of its peptide chains. Immunochemical studies show that an antiserum directed against one of the chains of luteinizing hormone crossreacts with both thyrotropin and luteinizing hormone, although, under the same conditions, each of the two hormones does not significantly cross-react with antisera prepared against the native form of the other. Data on the composition of the two chains of luteinizing hormone are given which confirm earlier observations of others that thyrotropin and one of the chains of luteinizing hormone have similar molar ratios of amino acids.

A method of pituitary-TSH (Thyroid-stimulating hormone) purification is applied to normal human placenta and allows the detection of a chorionic thyroid stimulating factor (Human-chorionic thyroid-stimulating factor, H.C. TSF).Immunological studies reveal a rather close but not complete antigenic relationship between human pituitary-TSH and H.C. TSF.Immunological data and the sustained biological effect into mice (test animal) suggest that H.C. TSF would possess some differences in structures when compared to human pituitary TSH.

SummaryA case of bilateral adrenocortical hyperplasia occurring in a case of metastatic carcinoma is reported.Clinically, the main features were : 1) acute generalized edema together with asthenia; 2) marked biochemical abnormalities (hypokalemia, alkalosis, hypo-chloraemia, hvpocalcaemia, diabetes); 3) exaggeration of pigmentation after adrenalectomy.From the endocrinological point of view : 1) increased Cortisol secretion rate; ox) raised plasma value of corticosterone; 3) increased aldosterone secretion rate; this latter fact seems to be an unusual feature of this syndrome and to play a minor role in pathogenesis of the biochemical abnormalities.

Resume L'examen clinique de 33 personnes d'une familletouchee par l'amyotrophie de Charcot-Marie-Tooth, nous a permis de decouvrir 8 cas evidents de cette affection. En outre, 6 cas sans manifestations cliniques n'ont ete deceles que par la mesure de la vitesse de conduction des nerfs noteurs. Chacun des 8 malades avait un pied creux indiscutable. Cinq des 6 cas latents presentaient un pied creux cliniquement evident. En outre, chez 14 sujets cliniquement et electriquement sains, on trouve 5 pieds creux reveles uniquement par la podoscopie. Nous en concluons que: 1.(1) La mesure de la conduction nerveuse permet d'etudier la diffusion familiale de la maladie de Charcot-Marie-Tooth, de maniere extremement sensible, en revelant les amyotrophies latentes. 2.(2) Il existe dans la famille L. P., un parallelisme frappant entre les deformations evidentes du pied (de type pied creux) et la maladie de Charcot-Marie-Tooth, visible ou latente. 3.(3) Un nombre relativement eleve d'individus sains est porteur d'un pied creux n'apparaissant qu'a la podoscopie. Leur conduction nerveuse est normale. Il faut considerer cette deformation isolee: soit comme un caractere independant de l'affection neuro-musculaire, soit comme le signe distal extremement precoce d'une maladie de Charcot-Marie-Tooth debutante. Seule, l'apparition eventuelle des signes de cette affection neurologique chez les individus porteurs d'un pied creux isole, ou dans leur descendance, permettra peut-etre de resoudre ce probleme.