Both in vivo FSH depletion and follicular exposure to Gonadotrophin-releasing hormone analogues in vitro are not effective to prevent follicular depletion during chemotherapy in mice.

STUDY QUESTION: Does fertility preservation using gonadotrophin-releasing hormone (GnRH) analogues during chemotherapy act through a direct effect on the ovary or through inhibition of FSH secretion? SUMMARY ANSWER: The absence of FSH in vivo and the direct exposition of ovarian follicles to GnRH analogues in vitro did not prevent chemotherapy-induced ovarian damage. WHAT IS KNOWN ALREADY: The potential mechanisms of action of GnRH analogues in protecting ovaries against chemotherapy damage remain poorly understood. We previously showed that GnRH analogues have a limited inhibitory effect on gonadotropin secretion and follicular growth in mice. STUDY DESIGN SIZE, DURATION: Mouse models were developed to independently evaluate (i) the indirect effect of FSH depletion on chemotherapy-induced ovarian damage using Fshb-deficient (-/-) mice to mimic the profound inhibition of FSH secretion during GnRH analogues treatment and (ii) the direct in vitro effect of GnRH agonist and antagonist in follicles exposed to chemotherapy using a follicular culture system. PARTICIPANTS/MATERIALS, SETTING, METHODS: To assess the indirect effect of GnRH analogues through FSH inhibition, Fshb-/- mice were treated with 1 IU pregnant mare serum gonadotropin (control group) or saline (study group) for 7 days and with cyclophosphamide (200 mg/kg) on Day 5. Ovaries were collected 48 h post-cyclophosphamide to evaluate ovarian reserve, cellular apoptosis and proliferation. To evaluate the direct effects of GnRH analogues on growing follicles, isolated preantral follicles from prepubertal mice were cultured in vitro for 13 days with 1 µM GnRH analogues and 20 µM of 4-hydroperoxycyclophosphamide or not at Day 4. Oocytes were matured by adding epidermal growth factor (EGF)/hCG on Day 12. Follicular development, follicular survival, oocyte maturation rates, cAMP production, and steroidogenesis were evaluated. To assess the direct GnRH analogues effects on follicular reserve, whole neonatal ovaries were cultured in vitro under the same conditions for 2 days. Ovaries were processed 24 h post-chemotherapy for ovarian reserve, cellular apoptosis and proliferation analysis. MAIN RESULTS AND THE ROLE OF CHANCE: Cyclophosphamide induced a significant follicular loss of more than 50% in Fshb-/- mice regardless of previous treatment with gonadotropins and no difference was observed in cell proliferation or apoptosis. In vitro experiments on growing follicles showed that 4-hydroperoxycyclophosphamide significantly decreased preantral follicle survival and maturation rates (55% and 37%, respectively) and delayed follicular development, regardless of the presence of GnRH analogues. Chemotherapy reduced granulosa cell numbers in all groups, while no change in cAMP production/106 granulosa cells was observed. Similarly, 4-hydroperoxycyclophosphamide induced apoptosis and significant follicular loss in cultured neonatal ovaries irrespective of GnRH analogues exposure. LIMITATIONS REASONS FOR CAUTION: As ovarian GnRH receptors expression differs in humans and mice, further studies are needed to validate our results in human ovaries. WIDER IMPLICATIONS OF THE FINDINGS: Our findings demonstrate that ovarian damage occurred even in the absence of FSH, suggesting that inhibition of the pituitary-gonadal axis is not involved in ovarian protection during GnRH analogues treatment. Using in vitro models, no evidence for direct protective effect of GnRH analogues against cyclophosphamide metabolite damage was observed. At present, clinical efficiency of GnRH analogues to prevent chemotherapy-induced ovarian damage remains highly debated and these experimental results reinforced the question as they did not bring evidence of direct or indirect mechanisms of protection. LARGE SCALE DATA: N/A. STUDY FUNDING AND COMPETING INTEREST(S): This work was supported by the Belgian FNRS, 'Le Fonds Emile DEFAY', and 'La Fondation Rose et Jean Hoguet'. Authors have no conflict of interest to declare.

Progesterone levels in letrozole associated controlled ovarian stimulation for fertility preservation in breast cancer patients.

STUDY QUESTION: Are progesterone levels after letrozole-associated controlled ovarian stimulation (COS) for fertility preservation in breast cancer patients, lower than after standard in vitro fertilization (IVF) cycles? SUMMARY ANSWER: During the luteal phase of letrozole-associated COS cycles (triggered with human chorionic gonadotrophin (hCG)) progesterone levels are similarly elevated to those obtained after standard COS without letrozole. WHAT IS KNOWN ALREADY: Current fertility preservation strategies for breast cancer patients include association of COS with the aromatase inhibitor letrozole to harvest several mature oocytes while maintaining low estradiol levels. Data on progesterone levels are however lacking despite growing evidence of the role of progesterone in breast tumorigenesis. STUDY DESIGN, SIZE, DURATION: This is a prospective observational study comparing estradiol and progesterone levels of 21 breast cancer patients undergoing letrozole-associated COS with 21 infertile patients undergoing standard COS for IVF and/or intra cytoplasmic sperm injection (ICSI). PARTICIPANTS/MATERIALS, SETTING, METHODS: All patients underwent COS with a GnRH antagonist protocol. In the fertility preservation group, ovulation induction was started in the follicular or luteal phase depending on the chemotherapy schedule and in 10 cases a GnRH antagonist was administered during luteal phase to induce luteolysis. Final oocyte maturation was induced by hCG in all patients. Estradiol and progesterone levels were measured on the day of hCG, at oocyte retrieval, and on days 3 and 8 after oocyte retrieval. Hormone levels in fertility preservation patients were compared with those observed in infertility patients. MAIN RESULTS AND THE ROLE OF CHANCE: While estradiol levels were significantly lower in the fertility preservation group compared with the control group (P < 0.001), progesterone levels were similar at all times, including patients receiving a GnRH antagonist during the luteal phase. LIMITATIONS, REASONS FOR CAUTION: The studied populations (breast cancer and infertile patients) are different, which may induce selection bias. The small sample size limits the study's statistical power and the possibility to perform multivariate analysis. Recruitment of the study and control patients was completed at the same time; however, enrollment of controls started at a later time. WIDER IMPLICATIONS OF THE FINDINGS: While the use of letrozole in fertility preservation patients has a favorable effect on estrogen levels, no benefit is seen for progesterone levels which are high and comparable with progesterone levels after standard COS in IVF patients. As progesterone has been associated with tumor cell proliferation, caution is mandatory. Modified protocols including GnRH agonist triggering should be investigated.

AMH mutations with reduced in vitro bioactivity are related to premature ovarian insufficiency.

STUDY QUESTION: Could anti-Müllerian hormone (AMH) mutations be implicated in the development of idiopathic premature ovarian insufficiency (POI)? SUMMARY ANSWER: Three rare or unknown missense variants of the AMH gene were identified in a cohort of 55 POI patients; all three variants showed a drastically reduced in vitro bioactivity. WHAT IS KNOWN ALREADY: Genetic factors are implicated in 5-15% of cases of POI. However, only a few genes have been shown to be involved in its development. AMH inhibits the recruitment of primordial follicles in the ovary and defective or absent AMH leads to premature depletion of the primordial follicle pool in AMH null mice. STUDY DESIGN, SIZE, DURATION: The whole coding sequence and the exon-intron junction of the AMH gene was sequenced in a cohort of 55 POI patients recruited over a period of 8 years. The studied variants were also sequenced in 197 ethnically matched controls. PARTICIPANTS/MATERIALS, SETTING, METHODS: POI was defined as amenorrhea of more than 4 months with increased FSH before the age of 40. Patients with POI resulting from radio- or chemotherapy, surgery, chromosomal anomalies or FMR1 gene pre-mutation were excluded from the study. Recombinant human wild-type (wt) and mutated AMH proteins were produced in HEK293 T cells. KK-1 cells transfected with the AMH receptor type 2 (AMHR2) and a BMP responsive element coupled to a luciferase reporter vector were stimulated with different concentrations of wt AMH and the three tested variants. MAIN RESULTS AND THE ROLE OF CHANCE: The whole coding sequence of the AMH gene could be performed and analyzed for 50 POI patients: 16 variants were found, including 6 missense variants from which 1 was unknown (R444H) and 2 were very rare (G264R and D288E). The variant D288E was also found in one of the patient's mother who also underwent POI at 32 years old. The stimulation of the AMHR2 assessed by the luciferase activity was drastically reduced for the three variants when compared with the wt AMH. LIMITATIONS, REASONS FOR CAUTION: The study is limited by a relatively small number of patients in the POI cohort. WIDER IMPLICATIONS OF THE FINDINGS: This is the first time that the bioactivity of AMH variants related to POI patients is tested in vitro. The functional study showed a drastic reduction of the protein activity for the three variants, supporting their contribution to the development of the ovarian insufficiency. The familial segregation further supports the implication of AMH in the development of POI. STUDY FUNDING/COMPETING INTERESTS: The study was performed thanks to funding from the 'Fondation Erasme'. No conflicts of interest are declared.

[Is ovarian function impaired in HIV patients? A clinical pilot study in Burkina Faso]. / La fonction ovarienne est-elle altérée chez les patientes séropositives pour le VIH? Une étude pilote au Burkina Faso.

A lot of studies published on the ten last years showed a decrease of fertility among HIV positive women. The present research aims to see if this decrease is linked to an ovarian failure, using AMH as principal marker of ovarian function. In this pilot study, 54 HIV-positive and 39 HIV-negative women were compared on the basis of their ovarian function, fecundity and possible ovarian failure. A blood sample was taken for hormonal titrations, HIV seropositivity, viral load and CD4 T cell count. An interview explored demographic characteristics, obstetrical and infectious history, and menstrual characteristics. This study was performed in Burkina Faso between January and February 2008. There is no significant difference after adjusting for age of AMH level between the two groups. However, in our study, 5.5% of HIV positive women had a premature menopause, which is a significant variation from the premature menopause rates of the African population, which is 1.4%. In conclusion, this study put the HIV impact on ovarian function into perspective but the high premature menopause rates could suggest an ovarian attack by the virus or the treatment.

[Anti-mullerian hormone and its role in the regulation of ovarian function. Review of the literature]. / Le rôle de l'hormone anti-muillerienne dans la régulation du fonctionnement de l'ovaire. Revue de la littérature.

Anti-mullerian hormone, also called AMH, belongs to the large family of transforming growth factor P. Its role in the sexual differentiation of male fetus is now well known. Recently, AMH has been demonstrated to play an important role in the ovarian function. In fact, AMH seems to regulate the kinetics of follicular development, inhibiting the follicular recruitment and the follicular growth. Thus, this intra-gonadic cybernin could be a decisive determinant of the rapidity of follicular pool exhaustion. Today, some experimental data from the literature suggest that AMH could be a reliable marker of ovarian reserve. This review summarizes the present knowledge about AMH and its role in physiology but also in ovarian pathology.

Impact of various endocrine and paracrine factors on in vitro culture of preantral follicles in rodents.

Folliculogenesis is a complex process regulated by various paracrine and autocrine factors. In vitro growth systems of primordial and preantral follicles have been developed for future use of immature oocytes, as sources of fertilizable oocytes and for studying follicular growth and oocyte maturation mechanisms. Rodents were often chosen for in vitro follicular culture research and a lot of factors implicated in folliculogenesis have been identified using this model. To date, the mouse is the only species in which the whole process of follicular growth, oocyte maturation, fertilization and embryo transfer into recipient females was successfully performed. However, the efficiency of in vitro culture systems must still be considerably improved. Within the follicle, numerous events affect cell proliferation and the acquisition of oocyte developmental competency in vitro, including interactions between the follicular cells and the oocyte, and the composition of the culture medium. Effects of the acting factors depend on the stage of follicle development, the culture system used and the species. This paper reviews the action of endocrine, paracrine factors and other components of culture medium on in vitro growth of preantral follicles in rodents.

Effect of insulin-like growth factor-I during preantral follicular culture on steroidogenesis, in vitro oocyte maturation, and embryo development in mice.

Insulin-like growth factor-I (IGF-I) is involved in the regulation of ovarian follicular development and has been shown to potentiate the FSH responsiveness of granulosa cells from preantral follicles. The aim of the present study was to investigate the effect of IGF-I during preantral follicular culture on steroidogenesis, subsequent oocyte maturation, fertilization, and embryo development in mice. Preantral follicles were isolated mechanically and cultured for 12 days in a simplified culture medium supplemented with 1% fetal calf serum, recombinant human FSH, transferrin, and selenium. In these conditions, follicles were able to grow and produce oocytes that could be matured and fertilized. The first experiment analyzed the effect of different concentrations of IGF-I (0, 10, 50, or 100 ng/ml) added to the culture medium on the follicular survival, steroidogenesis, and the oocyte maturation process. The presence of IGF-I during follicular growth increased the secretion of estradiol but had no effect on the subsequent oocyte survival and maturation rates. In the second experiment, IGF-I (0 or 50 ng/ml) was added to the culture medium during follicular growth, oocyte maturation, or both, and subsequent oocyte fertilization and embryo development rates were evaluated. Oocyte fertilization rates were comparable in the presence or absence of IGF-I. However, the blastocyst development rate was enhanced after follicular culture in the presence of IGF-I. Moreover, the total cell number of the blastocysts observed after differential labeling staining was also higher when follicles were cultured or matured in the presence of IGF-I.

Effect of preantral follicle isolation technique on in-vitro follicular growth, oocyte maturation and embryo development in mice.

BACKGROUND: The use of mechanical and enzymatic techniques to isolate preantral follicles before in-vitro culture has been previously described. The aim of this study was to assess the effect of the isolation procedure of mouse preantral follicles on their subsequent development in vitro. METHODS: Follicles were isolated either mechanically or enzymatically and cultured using an individual non-spherical culture system. Follicular development and steroidogenesis, oocyte in-vitro maturation and embryo development were assessed for both groups. RESULTS: After 12 days of culture, follicles isolated mechanically had a higher survival rate but a lower antral-like cavity formation rate than follicles isolated enzymatically. Enzymatic follicle isolation was associated with a higher production of testosterone and estradiol compared with mechanical isolation. A stronger phosphatase alkaline reaction was observed after enzymatic isolation, suggesting that follicles isolated enzymatically had more theca cells than those isolated mechanically. However, both isolation techniques resulted in similar oocyte maturation and embryo development rates. CONCLUSIONS: Enzymatic follicular isolation did not affect theca cell development. Follicular steroidogenesis was enhanced after enzymatic isolation but the developmental capacity of oocytes was comparable to that obtained after mechanical isolation.

[The medical chemistry department]. / Le Service de Chimie Médicale.

The laboratory of clinical chemistry performs more than 300 different tests in biochemistry, hormone and tumor markers analysis, therapeutic drug monitoring and toxicology. For the most basic tests it has followed the trend of clinical chemistry towards automation and since 2001 the heart of the laboratory is a modular automated system (MODULAR) including a preanalytical platform, unique in Belgium. For more sophisticated tests, the most recent techniques have been implemented, in particular capillary electrophoresis and ICP-MS ("inductively coupled plasma-mass spectrometry). Since 1994, the laboratory has become a reference center in the field of erythrocyte hereditary diseases, combining screening, diagnosis and research. The other research themes are the physiopathology of first trimester pregnancy and the P2Y receptors of extracellular nucleotides.

Persistent hyperparathyroidism requiring surgical treatment after kidney transplantation.

There are not many publications describing long-term follow-up of persistent hyperparathyroidism requiring surgical treatment after kidney transplantation (PHSKT). In some patients adenomas, rather than multiglandular disease, have been incriminated as the cause of PHSKT. We reviewed the charts of 45 patients followed for 12 to 146 months (median 45 months) after parathyroidectomy for PHSKT. We compared them with (1) those of 951 patients receiving a kidney graft during the same period but not submitted to parathyroidectomy or (2) 90 matched controls selected from this cohort to determine the characteristics of PHSKT patients. The duration of pretransplant dialysis was significantly longer in PHSKT patients than in controls (5.78 +/- 0.41 vs. 3.41 +/- 0.24 years; p < 0.0001). A total of 166 glands were removed or biopsied. Except for one questionable case, no true adenoma was observed even when only one gland was enlarged. The outcome of surgery was not influenced by the technique (subtotal parathyroidectomy versus total parathyroidectomy and autografting) but depended on the amount of resected parathyroid tissue: no failures and 4 cases of hypoparathyroidism in 34 cases with no missing gland at cervical exploration; 3 failures and no permanent hypoparathyroidism in 11 cases with one or two missing glands. Excision of the enlarged glands only was sufficient to cure the patient. No recurrence was observed. Our results suggest that single gland enlargement in PHSKT results in most cases from different rates of involution of the parathyroids after successful kidney transplantation. When fewer than four glands are discovered, resection of all visible glands with or without grafting corrects hypercalcemia in more than 70% of the cases.

Long-term results of subcutaneous parathyroid grafts in uremic patients.

HYPOTHESIS: Parathyroid glands are normally surrounded (entirely or partially) by fatty tissue. Subcutaneous parathyroid grafts are thus located in a normal environment. Therefore, we postulated that the late results of subcutaneous implantation of parathyroid tissue in uremic patients should be at least as good as those reported for intramuscular grafting. We also challenged the idea that the recurrence rate of renal hyperparathyroidism after surgery depended solely on the type of hyperplasia (diffuse vs nodular) observed in the implanted tissue. DESIGN: A retrospective study of a series of patients without loss to follow-up. SETTING: A university hospital and 9 affiliated dialysis units. PATIENTS AND INTERVENTIONS: Fifty-nine patients (33 women and 26 men) operated on for renal hyperparathyroidism underwent the resection of at least 4 parathyroid glands followed by presternal subcutaneous implantation of parathyroid tissue. They were followed up for 12 to 130 months (median, 38 months). MAIN OUTCOME MEASURES: Failure of treatment, recurrence of disease, and hypoparathyroidism. RESULTS: During the study period, 9 patients had to undergo another operation: 2 (3%) for persistent hyperparathyroidism due to a fifth ectopic gland and 7 (12%) for recurrence of hyperparathyroidism resulting from hypertrophy of the subcutaneous grafts. Four patients received a kidney transplant. The prevalence of hypoparathyroidism (intact parathyroid hormone serum level <1.6 pmol/L with a normal or low serum calcium concentration) was 14% (8 of 59 patients), and the curve representing the distribution of intact parathyroid hormone serum concentrations among operated on patients was shifted to the left when compared with the curve of patients who underwent hemodialysis and who had no indication for parathyroid surgery. In this latter group, the peak of the curve was situated between 1 and 2 times the upper normal limit, while it was in the normal range 12 to 130 months after total parathyroidectomy and subcutaneous parathyroid autotransplantation. No relation was observed between the recurrence rate of the disease and the histological characteristics of the parathyroid grafts. Also, their function was not influenced by the presence or absence of aluminum deposits in bone biopsy specimens that were obtained at the time of cervical exploration. CONCLUSIONS: The late results of total parathyroidectomy and presternal subcutaneous grafting compare favorably with the published data on other surgical techniques proposed for the treatment of renal hyperparathyroidism. The ease with which the hypertrophied grafts are removed when the disease recurs warrants further use of this procedure.

Blood biochemistry and endocrinology in the human fetus between 11 and 17 weeks of gestation.

Fetal blood was obtained in 35 normal pregnancies undergoing termination for psychosocial reasons between 11 and 17 weeks of gestation. Biochemical and endocrinological analyses were performed on each sample including concentrations of urea, creatinine, beta-microglobulin total protein, electrolytes, enzymes, alpha-fetoprotein (AFP), human chorionic gonadotrophin (HCG), thyroid stimulating hormone (TSH), thyroxin binding globulin (TBG), total thyroxin (TT4) and free thyroxin (FT4). The results were compared with values in maternal serum obtained at the same time. Fetal serum contained significantly higher concentrations of iron, beta-microglobulin, alkaline phosphatase (ALP), aspartate amniotransferase (AST), AFP, HCG and TSH and lower concentration of total protein, TBG and TT4 than maternal serum. Significant positive linear relationships were found between gestational age and the concentration of fetal serum total protein, ALP, TBG and FT4. Significant negative linear relationships were observed between gestational age and fetal serum beta-microglobulin and iron concentration. There were no significant correlations between fetal and maternal values. These data indicate that fetal blood biochemistry is not directly related to placental transfer and that the proteins and enzymes found inside the gestational sac are essentially of feto-placental origin with minimal contribution from the maternal protein metabolism. The comparison of coelomic fluid composition at 10-13 weeks with that of fetal serum at 11-17 weeks suggests that the anatomical changes in the human materno-fetal interface architecture between the early and late pregnancy periods could have a direct impact on materno-fetal transfer pathways.

Amniotic fluid biochemistry in second-trimester trisomic pregnancies: relationships to fetal organ maturation and dysfunction.

Protein and enzyme patterns were investigated in amniotic fluid samples of 12 trisomy 21 and five trisomy 18, between 11 and 16 weeks and of a control group of 50 chromosomally and anatomically normal pregnancies at the same gestational age. Positive correlations were found between gestational age and the concentration of urea (P<0.05), creatinine (P<0.01), beta2-microglobulin (P < 0.0001), hCG (P < 0.01), ALP (P < 0.001), LAP (P < 0.0001) and GGT (P <0.0005) in the amniotic fluid of euploid pregnancies. There were also significant correlations between the concentrations of the different variables measured except for hCG and AFP. Different protein and enzyme patterns were found in the amniotic fluid of aneuploid pregnancies where only beta2-microglobulin concentration was significantly correlated with advancing gestational age. The beta2-microglobulin concentration was significantly (P <0.05) higher in aneuploid fetuses presenting with hydrops compared to those without. These findings suggest that the changes in amniotic composition found during the fourth month of normal pregnancies reflect mainly the maturation of the fetal renal glomerular function and the arrival of enzymes from the digestive and respiratory tract into the amniotic cavity. In trisomy 21, there is evidence of a delay in the maturation of these organs whereas in trisomy 18, there is a reduced production of most proteins and enzymes.

Evaluation of surgical treatment of renal hyperparathyroidism by measuring intact parathormone blood levels on first postoperative day.

Intact parathormone (inPTH) has a short half-life. Its blood level on the first day after total parathyroidectomy and subcutaneous parathyroid implantation (PTX + G) should therefore allow an early diagnosis of missed residual parathyroid tissue. We tested this hypothesis in 72 uremic patients who were followed for 6 to 110 months after operation. Nine were reoperated for recurrence of the disease. Graft removal was successful in four patients who had post-PTX inPTH levels of 16 pg/ml or lower. In five patients, an overlooked parathyroid gland had to be resected. All of them had elevated post-PTX inPTH blood levels ranging from 72 to 791 pg/ml (upper normal limit 55 pg/ml). Three of these patients had presented with hypocalcemia after PTX. We conclude that the inPTH blood concentration on the first day after PTX allows more precise evaluation of the efficacy of the surgical procedure than the postoperative evolution of blood calcium levels. It is also useful for localizing the source of excessive PTH secretion (graft or overlooked gland) when the disease recurs.

Moderate doses of iodide in vivo inhibit cell proliferation and the expression of thyroperoxidase and Na+/I- symporter mRNAs in dog thyroid.

The function and the growth of adult thyroid gland is controlled by the opposite actions of thyrotropin (TSH) and iodide, the main substrate of the gland. Iodide deprivation leads to stimulation of the thyroid, improving the efficiency of iodide transport for hormone biosynthesis. We have investigated cell proliferation and thyroid specific gene expression 24 and 48 h after administering KI to dogs previously treated with goitrogens and perchlorate. In the hypothyroid dogs T3 and T4 serum levels decreased from 53 +/- 4 to < 30 ng/dl and from 1.6 +/- 0.6 to < 1 microg/dl respectively; TSH concentration increased from 0.16 +/- 0.02 to 2.7 +/- 0.4 ng/ml. After a 24 h moderate KI treatment (300 microg KI/dog of +/- 10 kg) serum T3 concentrations rose higher than the initial normal values, while T4 concentrations increased to reach values equivalent to the normal level. The high TSH concentration did not change significantly. The hyperplasia of the chronically stimulated thyroid resulting from goitrogens/NaClO4 treatment was not modified by this short term treatment with KI. In contrast, KI decreased the weight of the total gland and the level of cell proliferation, as determined by the fraction of cells incorporating BrdU. The effect of acute administration of KI on the expression of four major thyroid genes, the TSH receptor (TSHr), thyroglobulin (Tg), thyroperoxidase (TPO), and Na+/I- symporter (NIS) was analyzed by Northern blot. Tg, TPO and NIS mRNA expressions were up-regulated by chronic stimulation. The expression of the mRNAs of TSHr and Tg did not significantly differ between hyperstimulated and KI-treated dogs while TPO and NIS mRNA expression decreased after a 48 h KI treatment. TPO and NIS are therefore the only of these four genes whose expression is acutely modulated by iodide in vivo. Under TSH stimulation low doses of iodide resulted in: (1) decreased cell proliferation, (2) reestablished synthesis and secretion of thyroid hormones, (3) diminished TPO and NIS mRNA expression. Notably low doses of iodide under the same conditions had no effect on Tg and TSHr mRNA expression.

Selenium status in pregnant women of a rural population (Zaire) in relationship to iodine deficiency.

Endemic myxoedematous cretinism has been associated with combined selenium and iodine deficiency in several areas of Zaire. To determine selenium and iodine status across the country, serum selenium and thyroid function parameters including urinary iodide were determined at prenatal clinics in 30 health centres of rural villages distributed over the whole country. Only in Bas-Zaire was the mean serum selenium level similar to that in non-deficient areas (80-120 ng/ml); in the regions of Bandunda and Kasai levels were marginally decreased (55-80 ng/ml), and in Kivu, Haut-Zaire, Equateur and Shaba they were marginally or moderately decreased (< 55 ng/ml). The frequency of abnormally low urinary iodide (< 5 micrograms/dl) varied from 20% in the region of Bas-Zaire to 50% in Kasai (P < 0.001), and to still higher percentages in the 5 other regions of Zaire (Bandundu, 57%; Kivu, 63%; Equateur, 72%; Shaba, 76%; Haut-Zaire, 84%). With the exception of Bas-Zaire, biochemical maternal hypothyroidism (serum TSH > 5mU/l) was present in every region, with a frequency ranging from 3% in Kivu to 12% in Equateur. Iodine deficiency affects most of the Zairean population and requires public health measures on a larger scale than previously estimated. Combined iodine and selenium deficiency affects Equateur, Haut-Zaire and Kivu, where endemic myxoedematous cretinism occurs, but also Shaba, where it was not previously described. Besides combined iodine and selenium deficiency which is permissive, another factor (thiocyanate?) must be taken into account to explain the peculiarly elevated prevalence of endemic myxoedematous cretinism in Central Africa.

Long-term results after surgical treatment of renal hyperparathyroidism when fewer than four glands are identified at operation.

BACKGROUND: When fewer than four parathyroid glands are discovered during cervical exploration in cases of renal hyperparathyroidism, some have suggested that the glands could be resected and that no parathyroid tissue should be grafted. The aim of the present study was to evaluate this approach. Indeed, no detailed follow-up of such patients has been reported so far. STUDY DESIGN: Between September 1979 and July 1995, 157 patients underwent a cervical exploration for renal hyperparathyroidism in our department. In 23 cases, fewer than four parathyroid glands were found. The present study reports the results of 16 of these patients who did not undergo autotransplantation with parathyroid tissue after resection of all identified glands. RESULTS: At their last visit, four patients had normal blood levels of intact parathyroid hormone 14 to 71 months (median 60 months) after parathyroidectomy. The parathyroid hormone was not detectable in 2 patients at 16 and 76 months after operation. It was elevated in the 10 remaining patients, but the disease required no treatment or could be controlled medically in 7 patients with a follow-up ranging from 12 to 100 months (median 33 months). Two patients needed reoperation, one at 14 and one at 45 months after the first operation. An ectopic gland was found in both cases. The last patient refused further surgical treatment and died after 13 months. CONCLUSIONS: In most cases, autotransplantation of parathyroid tissue is not necessary when fewer than four glands are identified during parathyroidectomy. When grafting was omitted, 14 of 16 patients had normal or elevated blood levels of intact parathyroid hormone 10 to 100 months after parathyroidectomy (median 45 months). However, in two patients, intact parathyroid hormone was persistently undetectable for 16 to 76 months. This suggests that parathyroid tissue should be cryopreserved for delayed autotransplantation when the surgeon chooses not to graft parathyroid tissue.

Biochemical investigation of fetal renal maturation in early pregnancy.

The objective of this study was to evaluate the biochemical indices of normal fetal kidney maturation in early pregnancy. Urea, creatinine, gamma-glutamyltransferase, and beta 2-microglobulin levels were measured on paired samples of amniotic and coelomic fluids and maternal serum collected at the time of pregnancy termination in a group of woman with pregnancies between 8 and 11 wk, or on paired samples of amniotic fluid and maternal serum collected from woman undergoing early transabdominal amniocentesis between 11 and 16 wk. Before 11 wk of gestation (n = 12), significantly lower concentrations of creatinine and beta 2-microglobulin, and higher concentration of gamma-glutamyltransferase were found in amniotic fluid compared with both maternal serum and coelomic fluid. Significant positive correlations were found between gestational age (8-16 wk) and amniotic fluid (n = 47) levels of urea (r = 0.45; p < 0.01), creatinine (r = 0.68; p < 0.001), gamma-glutamyltransferase (r = 0.71; p < 0.001), and beta 2-microglobulin (r = 0.69; p < 0.001). The maternal serum levels of the corresponding variables did not varied significantly. The abrupt increase in creatinine, gamma-glutamyltransferase, and beta 2-microglobulin amniotic fluid concentrations after 10 wk of gestation provides biochemical evidence for the maturation of the fetal renal glomerular function at a time when the reabsorption capacity of the proximal tubular cells is not established. We suggest that this important variation in the amniotic fluid composition, unrelated to any comparable changes in the maternal serum, reflects the fetal kidney development from the mesonephros to the metanephros.

Hypothyroid patients showing shortened responsiveness to oral iodized oil have paradoxically low serum thyroglobulin and low thyroid reserve. Thyroglobulin/thyrotropin ratio as a measure of thyroid damage.

In Central Africa, all of northern Zaire is very severely deficient in iodine. A peculiar feature of this endemia is that iodine deficiency and the ensuing thyroid gland stimulation not only leads to goitre formation but also to progressive thyroid involution and to myxoedematous cretinism. An iodine supplementation trial based on oral administration of small doses of iodine was made in 81 schoolchildren. All of them received a small dose of iodine (0.1 ml containing 48 mg) per os and the thyroid status was followed during 4 months. Blood and urine samples were collected at the start of the study, then 2 weeks, 2 months and 4 months after iodine administration. Before iodine supplementation the mean urinary iodine level was 0.18 +/- 0.02 micromol/l, and 10% of the subjects had a urinary iodine level below 0.08 micromol/l. Fifty-two percent of the subjects had a serum thyrotropin (TSH) level above 10 mU/l. All the subjects responded to the administration of iodine. and all of them recovered a euthyroid status. Most of them were still euthyroid at the end of the study. However. within 4 or even 2 months, some subjects (15 % of the total) reverted to hypothyroidism. At the entry of the study these subjects were all hypothyroid and had elevated TSH and paradoxically low serum thyroglobulin (TG) values. In myxoedematous cretins living in the same area, even lower serum TG levels were found. Together with the absence of goitre, a paradoxically low serum TG Suggests a low thyroid reserve, and in the present case a reduced amount of functional thyroid tissue. We show that the serum TG/TSH ratio may be used as a predictive index of thyroid reserve and of positive response to iodine administration. These data further suggest that thyroid damage is not confined to myxoedematous cretins. but is widely distributed in the phenotypically normal population. Widely distributed thyroid damage may render iodine prophylaxis based on oral administration unpredictable.

The long-acting gonadotropin-releasing hormone analogues impaired the implantation rate.

OBJECTIVES: To determine the efficacy and innocuousness of long-acting versus short-acting GnRH analogues (GnRH-a) in long protocol for in IVF-ET. DESIGN: Prospective randomized study. SETTING: The IVF unit at an academic hospital. PATIENTS: One hundred couples admitted for their first IVF-ET attempt. MAIN OUTCOME MEASURES: Serum concentrations of LH, E2, and P during the all cycles and duration of pituitary desensitization were assessed, as well as fertilization rate, embryo quality, and implantation and pregnancy rates. RESULTS: Significantly more days (10.8 +/- 1.8 versus 9.2 +/- 1.7 days) of stimulation and more ampules of hMG (47 +/- 22 versus 33 +/- 16) were necessary to obtain similar numbers of embryos of quality with the long-acting GnRH-a. Implantation and delivery rates were significantly lower with the long-acting GnRH-a (32.8% versus 21.1%; 48.9% versus 29.1%, respectively). CONCLUSIONS: As the long-acting GnRH-a might interfere with the luteal phase and embryo development, short-acting GnRH-a should be preferred for ovarian hyperstimulation in IVF-ET.