Introduction: Endocrine gland-derived vascular endothelial growth factor (EG- VEGF) was recently identified as the first tissue-specific angiogenic molecule. EG-VEGF is expressed in tissues such as the adrenal glands, testis, ovary and placenta. Objective: The aim of this study was to investigate the EG-VEGF expression and regulation by testosterone in rat pancreatic tissue. Material and Methods: Samples were obtained from 25 adult male wistar rats, divided in 5 groups: 1) control, 2) gonadectomized, 3, 4 and 5 gonadectomized and substituted with testosterone enantate for 3, 5 and 7 days, respectively. Serum hormone concentrations were studied by RIA. Total RNA was isolated from pancreas, adrenals glands, and testis. EG-VEGF expression was studied by real-time PCR. EG-VEGF goat polyclonal antibody was used to determine the cellular localization of the protein in pancreatic tissue. Results: EG-VEGF expression was detected in all pancreatic samples, with higher serum testosterone concentrations (groups 3 and 4) presented lower EG-VEGF expression. All samples examined showed specific immunostaining for EG- VEGF. The positive labeling was localized within the pancreatic islets. Conclusion: The transcriptional activity of the EG-VEGF gene in the pancreas was modulated by the androgenic milieu, suggesting that testosterone is one of the factors involved in pancreatic EG-VEGF regulation. These results are in accordance with other studies in different endocrine tissues, such as the ovary and adrenal.

Introduction: Endocrine gland-derived vascular endothelial growth factor (EG-VEGF), and the mammalian homologue of Bombina variegata peptide 8 (Bv8), are two highly related endothelial cell mitogens and chemoattractant factors with restricted expression profiles. Human EG-VEGF messenger RNA is expressed by steroid producing cells in the adrenal gland, placenta, testis and ovary. Consistent with such expression pattern, the human EG-VEGF gene promoter has a potential binding site for steroidogenic factor SF-1, a pivotal element for steroidogenic-specific transcription. The occurrence of SF-1 in the endothelial cells of the blood vessels together with the steroidogenic properties of the pancreas are two characteristics that might account for the possibility that EG-VEGF could be the regulatory factor in angiogenesis in this tissue. The aim of this study was to investigate the expression and localization of the EG-VEGF in normal human pancreas and pancreatic adenocarcinoma. Methods: Total RNA was extracted from male (5) and female (5) samples, obtained from the organs donors program and samples (2) of surgically resected adenocarcinoma. The relative expression of pancreatic EG-VEGF was assessed by RT-PCR analysis using specific primers. Anti-EG-VEGF goat polyclonal Ig-G (sc-30343 Santa-Cruz) antibodies were used to determine the cellular localization of the protein in pancreatic tissue. Results: By RT-PCR, specific amplicons for EG-VEGF were detected in both male and female samples, though a differential expression was observed. According to densitometric analysis, the relative abundance of mRNAs was: female adenocarcinoma 0.89> female normal pancreas 0.64> male adenocarcinoma 0.71> male normal pancreas 0.38. Immunohistochemical studies also showed a different staining. In male samples, the signal was localized preferentially in the pancreatic islets while in female samples, the main staining was observed towards the exocrine portion. In tumoral samples, the specific staining showed a diffuse localization pattern. Conclusion: The expression of EG-VEGF/PROK1 gene in pancreas resembles to that reported for other endocrine tissues, such as the ovary and adrenals and adds evidence to the linkage of steroids with pancreatic physiology. These results also demonstrate a differential expression between normal and tumoral tissues. The human pancreas expresses EG-VEGF, a highly specific mitogen, which regulates proliferation and differentiation of the vascular endothelium in a tissue-specific manner.

Orphan nuclear receptor steroidogenic factor-1 (SF-1) is crucial for development and function of steroidogenic organs. The steroidogenic factor-2 (SF-2) is an essential factor involved in cholesterol transfer and activation of promoters of steroidogenic enzymes CYP11A1, CYP17 and Steroidogenic Acute Regulatory Protein (StAR). We have previously demonstrated steroidogenic activity in pancreatic tissue. The aim of this study was to investigate the presence of SF-1 and SF-2 in human pancreas. Total RNA was extracted from normal male (five) and female (five) samples, obtained from the organs donor program. RT-PCR approach was used to analyze the expression of SF-1 and SF-2. Immunohistochemical analysis was performed for SF-1. The bands of expression were present in both male and female samples, although differential expression was observed. For both factors, the signal detected was more evident in males than in females. A similar pattern was present in the immunohistochemical study. Normal human pancreas expresses SF-1 and SF-2 factors similarly to ovary and adrenals. A distinctive characteristic is the sexually dimorphic expression of these factors. Our data provide evidence suggesting that the pancreas achieves steroidogenic activity supporting the presence of gender- and location-related differences in the expression of these steroidogenic factors.

INTRODUCTION:Papillary cystic neoplasm (PCN) of the pancreas is a low-malignancy tumor affecting predominantly young females. Sex steroid hormones have been involved in its development and/or growth. Estrogen receptor (ER) has been scarcely found in this tumor, although there is some evidence suggesting expression of the beta-isoform. Unlike ER, progesterone receptor (PR) expression has been consistently observed. Immunohistochemical analysis of the two isoforms of ER has not been performed in this tumor.AIM:To characterize expression of ER isoforms with an immunohistochemical method.METHODOLOGY:Expression of ER-alpha, ER-beta, and PR was analyzed by immunohistochemistry using isoform-specific ER and PR antibodies in paraffin-embedded tissue blocks from seven cases of PCN of the pancreas.RESULTS:Most patients were young females. ER-alpha and ER-beta were present in two and six tumors, respectively. PR was identified in six tumors.CONCLUSIONS:ER-beta expression predominates over the alpha-isoform in PCN of the pancreas. This finding supports the idea that previous negative results on ER expression were a consequence of the use of antibodies with no anti-beta activity. The role of ER-beta in the milieu of factors promoting the development and aggressiveness of PCN needs to be elucidated to address novel diagnostic and therapeutic approaches.

Coexistence of hyperinsulinemia and hyperandrogenism in women has been frequently described. Most of the studies addressing this issue have focused on the mechanisms by which insulin produces hyperandrogenism. In the present study, we analyzed the effects of testosterone in vivo and in vitro upon insulin gene expression and release in the rat. Our studies demonstrate that testosterone increases insulin messenger RNA (mRNA) levels in vitro as well as in vivo. In both prepuberal and intact adult rats, serum testosterone concentrations were positively correlated with insulin mRNA levels and insulin concentration in serum. Testosterone deprivation after gonadectomy decreased both insulin gene expression and serum insulin concentration. Insulin mRNA levels were partially restored after 3 days of testosterone administration and serum insulin was 80% and 27% above baseline values at 5 and 7 days posttreatment. Primary cultured pancreatic islets treated with the sexual steroid increased about 80% insulin mRNA, as well as protein, and release. In transfected islets, testosterone increased the activity of the -410 bp rat insulin promoter I by 154%. These data demonstrate that testosterone has a direct effect upon pancreatic islet function by favoring insulin gene expression and release.

After the International Conference on Population and Development, held in Cairo in 1994, Mexico took the initiative to implement a Reproductive Health Programme as part of the reform process of the public health sector. Creation of the General Directorate of Reproductive Health and the Inter-institutional Group of Reproductive Health and elaboration of a comprehensive programme were the first actions undertaken. Different actions relating to the main components of the programme family planning, perinatal health and women's health were carried out including interpersonal communication techniques and incorporation of gender perspectives. These actions have been reviewed in this paper. Strategies for the implementation of the programme such as training, production of media materials, "Mother and Baby-Friendly Hospital" initiative, epidemiological surveillance systems, and actions aimed at improving quality of services, have been highlighted. Some statistical data on achievements of the programme have been presented as the evolution of prevalence of contraceptive use, total fecundity rate, maternal and perinatal mortality and life expectancy, reflecting that the implementation of the programme has been successful. However, a number of outstanding needs are still outstanding and new demands have emerged.

Sex steroid hormones influence insulin homeostasis and glucose metabolism, estradiol (E2) and progesterone (P4) induce changes in both fasting and postprandial insulinemia in rodents, however, insulin gene expression during estrous cycle is unknown. The aim of the present study was to determine an insulin gene expression pattern during the estrous cycle in the rat. Groups of 6 adult rats in each day of the estrous cycle were used. Serum P4, E2, testosterone (T) and insulin concentrations were determined by radioimmunoassay (RIA). A Northern blot analysis was performed to assess insulin gene expression in pancreatic tissue. We found a marked variation in insulin gene expression during the estrous cycle. The highest insulin expression was observed during the proestrus day. Interestingly, E2 and P4 but not T levels were correlated with changes in insulin mRNA content. The variations in serum insulin during the cycle were correlated with its mRNA content in pancreas. The overall results showed variations in serum insulin and insulin gene expression during estrous cycle of the rat that correlated with circulating E2 and P4 levels.

OBJECTIVE:To assess the serum concentrations of estradiol (E2), estrone (E1), gonadotrophins, sex hormone-binding globulin, and lipids, and to determine degree of symptom relief after subcutaneous implantation of 25 mg estradiol in postmenopausal Mexican women.DESIGN:Fifteen postmenopausal, hysterectomized women participated in an open, observational study. Blood samples were obtained before implantation and at regular intervals during a study period of 24 weeks. Climacteric symptoms were evaluated by means of the Greene climacteric scale. Wilcoxon's test was performed on the paired results of pre-and postimplantation values.RESULTS:Serum concentrations of E2 obtained after implantation were fairly constant, remaining within the early follicular range for the entire study period of 24 weeks, and were associated with significant symptom relief. A physiological, premenopausal E2:E1 ratio was achieved. No significant metabolic changes occurred. Side effects were estrogenic in nature and no removal of implant was required.CONCLUSIONS:Subcutaneous implantation of 25 mg estradiol results in physiological, premenopausal estrogen concentrations in most women and is associated with considerable symptom relief without inducing significant adverse metabolic effects.

In developing countries, health problems such as malnutrition and anaemia, unwanted pregnancy, sexual and gender violence, unregulated fertility, reproductive tract cancers and osteoporosis contribute to women's ill health throughout their reproductive life cycle. Further about one-third of the total disease burden in women between 15 and 44 years of age is linked to health problems related to pregnancy. Although the priorities for specific reproductive health services differ from country to country, the identification, understanding and prevention of reproductive health problems affecting local and regional populations is mandatory for the decision making and the implementation of problem solving programs of action. The establishment of reproductive health and research priorities as a result of needs assessment, is the best strategy for implementing specific projects focused on national needs and priorities as well as on issues of global interest. Human resource development in research is the key element and has to be understood as a system that guarantees the continuity of efforts in improving the reproductive health status of the population. The improvement of the reproductive health of population can be accomplished through the active participation of a "critical mass" of researchers in reproductive health and a strong commitment at the national level for implementing and applying results in practice.

Background The present study was carried out to investigate the functional significance of the reduced dopaminergic tone in subjects affected with polycystic ovary syndrome (PCOS).Methods. Our group evaluated he response of pituitary PRL, LH, FSH, and TSH to the administration of a single 10-mg oral dose of the dopamine (DA) receptor antagonist metoclopramide in lean (n = 7) and obese (n = 8) PCOS women and in 11 regularly cycling age- and weight-matched controls (six lean and five obese). In addition, circulating PRL bioactivity was evaluated by its mitogenic activity on a lymphoma cell bioassay.Results. Oral administration of metoclopramide resulted in a significant increase in serum PRL in all subjects; however, the highest increments, regardless of body mass index (BMI), were observed in control women (p <0.005). Measurements of PRL mitogenic activity on the Nb2 lymphoma cell bioassay revealed a significant increase in the bioactive/ immunoreactive (B/I) ratio of PRL under basal and stimulated conditions in obese PCOS subjects (p <0.05), Mean fasting glucose/insulin and glucose/insulin-AUC ratios were significantly lower (p <0.001) in obese PCOS when compared with all other groups.Conclusions. These data support the existence of low DA hypothalamic tone in PCOS women that is likely involved in the inappropriate LH and PRL secretion frequently seen in this syndrome, In addition, our results suggest changes in PRL bioactivity in obese PCOS that may play a role in the development of hyperinsulinemia; however, whether PRL has a functional significance in the development of the metabolic disturbances frequently seen in PCOS remains to be elucidated. (C) 2000 IMSS, Published by Elsevier Science Inc.

The immunoreactivity of various LH and FSH calibration standards and recombinant preparations in the enzyme-linked immunoassay (EIA) systems for gonadotrophins developed for the Special Programme of Research in Human Reproduction of the World Health Organization (WHO) were compared. The preparations tested included three LH and two FSH pituitary standards (calibrated against LH 80/552 and 68/40 and FSH 78/549 respectively) provided with the EIA or radioimmunoassay WHO matched reagent kits, the pituitary preparation LER-907, and recombinant human LH (rhLH) and FSH (rhFSH). Simultaneous curve fitting of the EIA dose-response curves revealed no significant differences among the slopes generated by the WHO LH standards and LER-907; in contrast, no parallelism was found between the curves of rhLH and the pituitary-derived LH standards. No significant differences were found among the slopes of the curves elicited by the pituitary and recombinant FSH preparations. Each LH preparation exhibited a high degree of charge heterogeneity. Considerable variations in charge isoform distribution among the WHO LH standards, rhLH and LER-907 were also evident. In contrast, the FSH preparations were less heterogeneous and exhibited minor differences in charge distribution. Despite the existing differences in charge isoform distribution, all the pituitary-derived preparations as well as rhFSH seem appropriate for using as calibration standards in this particular EIA system.

In pancreas, the activities of several sex steroid-transforming enzymes have been reported. Data have been obtained in perfused organs, total tissue homogenates, and subcellular organelles. These data, concurrent with the description of the presence of ligand-regulated steroid receptors, as well as the sexually dimorphic behavior of some pancreatic tumors, are clear evidence in support of the participation of steroid hormones in the pancreatic function. In this study, the steroidogenic ability of the pancreas was demonstrated by two different methods: (a) in tissue homogenates, by the identification of cytochrome P-450scc gene (CYP11A) transcripts after reverse transcription-polymerase chain reaction amplification (RT-PCR); and (b) in isolated mitochondria by the glutethimide-dependent inhibition of cholesterol-pregnenolone biotransformation. The results obtained in a series of independent experiments showed that (a) the pancreatic tissue possessed transcriptional activity of the CYP11A gene, although to a lesser extent than the typical steroidogenic tissues, and (b) isolated mitochondria obtained from the pancreas were able consistently to synthesize pregnenolone; furthermore, the addition of the specific inhibitor aminoglutethimide (AMG) blocked its synthesis. On the whole, these findings are interpreted as clear evidences of the activity of the cytochrome P-450scc enzymatic complex (P450scc), responsible for the transformation of cholesterol into pregnenolone and considered the first and limiting step in steroid biosynthesis.

CONCLUSIONThe rate of growth of a papillary-cystic tumor of the pancreas seemed to be enhanced by the concurrence of pregnancy. Progesterone receptors in the tumor were demonstrated by immunohistochemistry and by molecular biology methods.BACKGROUNDPapillary cystic tumor of the pancreas is extremely rare, occurring predominantly young females. Owing to the low frequency of the tumor, there is scarce information about the conditions that promote tumor growth.METHODSIn this report, we present the temporal association between very rapid growth of a papillary-cystic neoplasm and pregnancy. Clinicopathological, immunohistochemical, and molecular biology analyses were performed.RESULTSA 21-yr-old woman was admitted because of recurrent epigastric abdominal pain associated with episodes of nausea and vomiting, and a history of an abdominal tumor of about 50 mm near the head of the pancreas, detected by ultrasound. On admission the patient had a flat, nontender abdomen without palpable masses. Laboratory analysis including hematologic and hepatic tests were strictly normal; only CA 19-9 (42 U/mL, normal 37 U/mL) was above normal values. One week after admission, an abdominal computerized axial tomography (CAT) scan revealed an 81.6-mm cystic mass localized in the head of the pancreas, and 1 wk later, in a laparotomy, a papillary-cystic neoplasm of 120 mm, limited to the head of the pancreas, was found. Three months later, in a routine follow-up visit, an 18-wk pregnancy was clinically diagnosed and confirmed by ultrasound exploration. The pregnancy continued without complications, and a normal male infant (3.7 kg) was born at 39 wk of gestation, by vaginal delivery. Eighteen months after tumor resection, the patient was asymptomatic and her child was in good health. We propose that progesterone affects tumor growth.

In true hermaphroditism diverse phenotypes and karyotypes are found; there are no distinctive laboratory features that can distinguish it from other intersex disorders, thus the diagnosis is made by the histological findings. Existence of Leydig cells is demonstrated by testosterone levels above the female range; however, presence of ovarian tissue cannot be ascertained because of the absence of a reliable functional test. Unless appropriate biopsies are performed or the whole gonad is removed, there is a risk of not diagnosing true hermaphroditism. To find a reliable test that can differentiate patients with true hermaphroditism from those with other intersex disorders, we investigated the estradiol (E-2) response to human menopausal gonadotropins (hMG) in infants with genital ambiguity. These results were correlated with the histological findings.Eleven infants with genital ambiguity and four with a high scrotal testis were stimulated every 12 h with 2 IU/kg hMG. If E-2, rose above 80 pg/mL (cut-off point), the test was discontinued; if after 7 days E-2, remained below 80 pg/mL, the hMG dose was doubled and stimulation extended for 7 additional days. In five patients in whom true hermaphroditism was later histologically demonstrated, E-2, rose above 80 pg/mL. In two of them, ovarian tissue was removed and hMG stimulation repeated; no response above our cut-off point was observed during the second test. The maximal E-2, response to hMG in the remaining 10 individuals was 43 pg/mL; after laparotomy or gonadal biopsies no ovarian tissue was found. The hMG stimulation test can be considered a reliable and safe dynamic procedure for demonstrating the presence or absence of ovarian tissue in infants with genital ambiguity.

Androgens influence the incidence and prevalence of pancreatic cancer in humans and animal models. To our knowledge there has been molecular demonstration of the presence of neither the androgen receptor (AR) nor transcripts of the AR gene. Reverse-transcription polymerase chain reaction (RT-PCR)-Southern blotting was employed for molecular detection and measurement of the androgen receptor messenger ribonucleic acid (AR mRNA) in pancreas. Total RNA obtained from pancreas, prostate, seminal vesicles, and testes of neonatal and adult male and female rats, as well as castrated males substituted with testosterone cyclopentylate, was analyzed by Northern blot technique. Positive hybridization to AR cDNA was obtained in all tissues assayed but not in the pancreas. However, a clear AR 32P cDNA hybridization signal was obtained in pancreatic tissues after cDNA synthesis using RT-PCR-Southern blotting. The levels of the AR transcripts obtained by RT-PCR in the various pancreatic samples were as follows: adult females and neonatal animals > castrated adult males > adult males > castrated adult males substituted with testosterone. These results indicated that the pancreatic tissue possessed transcriptional activity of the AR gene, although to a lesser extent than the typical androgen responsive tissues (prostate and seminal vesicles). In conclusion, transcriptional activity of the AR gene in the pancreas seemed to be modulated by the androgenic milieu in the tissue similar to that reported for the classical androgen-responsive organs.

PROBLEM:Experimental and clinical evidence has suggested an immunostimulatory effect of prolactin and that bromocriptine, an inhibitor of prolactin release, counteracts the actions of prolactin on the immune system. The aim of this study was to determine the impact of elevated serum prolactin levels on the immune system in patients with pathological hyperprolactinemia.METHOD:For this purpose, parameters of the cellular and humoral immune system were studied in six women with prolactinomas and one with idiopathic hyperprolactinemia. Studies were performed when serum prolactin concentrations were high as well as during different phases of the menstrual cycle when prolactin levels had been normalized through treatment with bromocriptine.RESULTS:Hyperprolactinemic subjects, when compared with six age-matched normal women, had significantly higher percentages of total lymphocytes and CD2+ cells. Elevation of CD4+ cells was also observed although to a lesser extent. Bromocriptine-treated patients, when compared with normal women were characterized by increased numbers of total lymphocytes and CD4+ cells, decreased percentage of CD8+ cells, and increased concentrations of serum IgM. These last two findings were also significantly different when compared to those observed in hyperprolactinemia.CONCLUSION:In this study we have described the changes on cellular and immune parameters in patients with hyperprolactinemia before and during bromocriptine therapy, which support the links of communications between the immune and endocrine systems in humans.

Objective. There has been increasing evidence on the mechanisms underlying the interactions between the neuroendocrine and the immune systems, particularly in animal models with relatively few information in the human. In this study, we evaluate the cellular and humoral immunity in female patients with hypopituitarism and in normal women throughout the menstrual cycle in an attempt to determine the role of pituitary and gonadal hormones on the immune system.Design. Serum immunoglobulins, peripheral blood lymphocyte subsets, and serum hormones were measured in eight patients with postpartum pituitary necrosis (Sheehan's syndrome) and in six normal women along different phases of the menstrual cycle, taking advantage of the lack of pituitary function and the cyclic variations in serum hormones, respectively.Results. Patients with Sheehan's syndrome had higher T lymphocytes (CD2), including helper (CD4) and suppressor (CD8) cell subpopulations and B lymphocytes (CD 19) when compared with normal menstruating women. An increase of serum IgA concentrations was also observed. Normal women showed little non-statistically different changes along the menstrual cycle in peripheral blood cell parameters and in serum immunoglobulin levels.Conclusions. a) Hypopituitarism in humans, in contrast with the animal model, may associate with immune up-regulation at both cellular and humoral levels; and b) hormonal changes along the normal menstrual cycle probably do not influence in great extent the immune system.

The effect of estradiol (E2) and progesterone (P4) on the IgA system, both at the systemic and at the mucosal level, was studied in 10 healthy young adult Mexican women during two consecutive menstrual cycles (MC); the control group consisted of five young adult Mexican men. Eight matched samples of blood and parotid saliva were obtained, in which serum E2, P4, and IgA were quantified. Parotid saliva was obtained with Curby's device and sIgA was quantified by an ELISA method. The MC was divided into follicular phase (FP, days 1 to 16) and luteal phase (LP, days 17 to 30). Serum IgA showed slight fluctuations along the period of study, but they were not different between women and men. Irrespective of the phase of the MC, salivary sIgA was higher in women than in men (P<0.01); sIgA was slightly but not significantly higher in the FP, as compared to the LP. The comparison of phases in each individual woman showed significantly higher levels in the FP (P<0.01). The profile of sIgA in saliva observed in women resembled the pattern of serum E2 (r = 0.859, P<0.05), suggesting a possible relation of E2 in the secretion of sIgA by the parotid gland.