Sample records for diethylstilbestrol

  1. Diethylstilbestrol-scaffold-based pregnane X receptor modulators.

    Hodnik, Žiga; Tomašič, Tihomir; Smodiš, Domen; D'Amore, Claudio; Mašič, Lucija Peterlin; Fiorucci, Stefano; Kikelj, Danijel


    Due to its function as a regulator of drug-metabolizing enzymes and transporters, pregnane X receptor (PXR) represents an important factor involved in drug metabolism. In this work, we describe the discovery of diethylstilbestrol-based PXR modulators, which were designed from marine sulfated steroids with PXR agonistic activity, solomonsterols A and B, and our recently reported bazedoxifene scaffold-derived PXR antagonists. The methylated diethylstilbestrol derivative 1 displayed potent PXR agonistic activity with an EC50 value of 10.5 μM, whereas compounds 3, 4 and 6 (IC50 for 6 = 27.4 μM) and diethylstilbestrol (2) itself (IC50 = 14.6 μM) exhibited PXR antagonistic effects in HepG2 cells. The PXR modulatory effects of the synthesized diethylstilbestrol derivatives were further confirmed by the induction of PXR-regulated CYP3A4 expression with compound 1, as well as by the inhibition of the rifaximin-promoted up-regulation of CYP3A4 expression with 2 and its derivative 6.

  2. Effect of diethylstilbestrol on polyamine metabolism in hamster epididymis

    Chun-HongQiu; MasatoOhe; ShigeruMatsuzaki


    Aim: To investigate the effect of diethylstilbestrol (DES), one of the most potent endocrine disruptors, on the metabolism of polyamines in hamster epididymis. Methods: Male golden hamsters of 7-week-old were kept under a light and dark cycle of 14 h and 10 h for 1 week to stimulate maximally the gonadal function. DES was injected subcutaneously at doses of 0.01mg·kg-1·day-1,0.1mg·kg-1·day-1 and 1mg·kg-1·day-1 for one week. Results:DES treatment caused a significant decrease in the weight of epididymis. The activity of epididymal ornithine decarboxylase (ODC) increased 1 day after DES treatment, kept at a high level for 4 days and then decreased to nearly normal level at day 7. The activity of spermidine/spermine N1-acetyltransferase (SSAT) also increased transiently after DES treatment. The contents of putrescine, spermidine, spermine and N1-acetylspermidine were increased 1 day 4 days after DES treatment and restored to normal at day 7. All these changes showed a marked difference between the caput and the cauda. Conclusion: The polyamine biosynthesis in the hamster epididymis can be affected by DES,a xenoestrogen. DES may probably affect polyamine metabolism in the epididymis by regulating the rate-limiting enzymes involved in the polyamine biosynthesis.

  3. Neonatal exposure to diethylstilbestrol causes granulomatous orchitis via epididymal inflammation.

    Miyaso, Hidenobu; Naito, Munekazu; Hirai, Shuichi; Matsuno, Yoshiharu; Komiyama, Masatoshi; Itoh, Masahiro; Mori, Chisato


    Diethylstilbestrol (DES), an endocrine-disrupting chemical, is an infamous artificial estrogenic compound. Although neonatal exposure to DES has been shown to result in inflammation of the male reproductive system, it has not, to our knowledge, been reported to induce testicular inflammation. Here we report that neonatal exposure to DES caused granulomatous orchitis with spermatogenic disturbance in 4 of 17 ICR male mice at 12 weeks of age. In the animals with spermatogenic disturbance, we observed either seminiferous tubules containing only cells with Sertoli cell features (likely Sertoli cell syndrome), or tubule cells in maturation arrest that contained only spermatogonia and/or spermatocytes. Following neonatal DES exposure, 5-week-old mice exhibited inflammation in cauda epididymis; by 8 weeks, the inflammation had spread to all segments of epididymis but not the testis; by 12 weeks, inflammation of the epididymis was observed in all mice. These data indicated that cauda epididymis has increased sensitivity to neonatal DES exposure compared to other segments of epididymis and testis. The data also implied that neonatal DES exposure-induced inflammation in cauda epididymis extended gradually to the testis via corpus and caput during development.

  4. Adrenal steroidogenesis disruption caused by HDL/cholesterol suppression in diethylstilbestrol-treated adult male rat.

    Haeno, Satoko; Maeda, Naoyuki; Yamaguchi, Kousuke; Sato, Michiko; Uto, Aika; Yokota, Hiroshi


    The synthetic estrogen diethylstilbestrol is used to prevent miscarriages and as a therapeutic treatment for prostate cancer, but it has been reported to have adverse effects on endocrine homeostasis. However, the toxicity mechanism is poorly understood. Recently, we reported that diethylstilbestrol impairs adrenal steroidogenesis via cholesterol insufficiency in adult male rats. In the present study, we found that the adrenal cholesterol level was significantly reduced without of the decrease in other precursors in the adrenal steroidogenesis 24 h after a single dose of diethylstilbestrol (0.33 μg/g body mass). The serum HDL/cholesterol level was also reduced only 12 h after the diethylstilbestrol exposure. The level of Apo E, which is indispensable for HDL/cholesterol maturation, was decreased in both the HDL and VLDL/LDL fractions, whereas the level of Apo A1, which is an essential constituent of HDL, was not altered in the HDL fraction. Because the liver is a major source of Apo E and Apo A1, the secretion rates of these proteins were examined using a liver perfusion experiment. The secretion rate of Apo A1 from the liver was consistent between DES-treated and control rats, but that of Apo E was comparatively suppressed in the DES-treated rats. The disruption of adrenal steroidogenesis by diethylstilbestrol was caused by a decrease in serum HDL/cholesterol, which is the main source of adrenal steroidogenesis, due to the inhibition of Apo E secretion from the liver.

  5. Diethylstilbestrol-induced mouse hypospadias: "window of susceptibility".

    Sinclair, Adriane Watkins; Cao, Mei; Baskin, Laurence; Cunha, Gerald R


    This review presents published and novel results that define the programming window for diethylstilbestrol (DES)-induced abnormal development of the mouse penis. These data indicate that DES has its greatest effect during the period of most intense penile morphogenesis, namely postnatal days 0-15 (P0-P15). Pregnant mice and their neonatal pups were injected subcutaneously with 200 ng/gbw DES every other day from embryonic day 12-18 (DES E12-E18), postnatal day 0-10 (DES P0-P10), embryonic day 12 to postnatal day 10 (DES E12-P10), postnatal day 5-15 (DES P5-P15), and postnatal day 10-20 (DES P10-P20). Aged-matched controls received sesame oil vehicle. After euthanasia at 10, 15, 20 and 60 days, penises were analyzed by gross morphology, histology and morphometry. Penises of all 5 groups of DES-treated mice were reduced in size, which was confirmed by morphometric analysis of internal penile structures. The most profound effects were seen in the DES E12-P10, DES P0-P10, and DES P5-P15 groups, thus defining a DES "programming window". For all parameters, DES treatment from P10 to P20 showed the most mild of effects. Adverse effects of DES on the MUMP cartilage and erectile bodies observed shortly after the last DES injection reverted to normality in the DES P5-P15, but not in the E12-P10 and P0-P10 groups, in which MUMP cartilage and erectile body malformations persisted into adulthood, again emphasizing a "window of susceptibility" in the early neonatal period.

  6. Quantitative proteomic determination of diethylstilbestrol action on prostate cancer

    Pierre Bigot; Kevin Mouzat; Souhil Lebdai; Muriel Bahut; Nora Benhabiles; Géraldine Cancel Tassin; Abdel-Rahmène Azzouzi


    Diethylstilbestrol (DES) has a direct cellular mechanism inhibition on prostate cancer.Its action is independent from the oestrogen receptors and is preserved after a first-line hormonal therapy.We aimed to identify proteins involved in the direct cellular inhibition effects of DES on prostate cancer.We used a clonogenic assay to establish the median lethal concentration of DES on 22RV1 cells.22RV1 cells were exposed to standard and DES-enriched medium.After extraction,protein expression levels were obtained by two-dimensional differential in-gel electrophoresis (2D-DIGE) and isotope labelling tags for relative and absolute quantification (iTRAQ).Proteins of interest were analysed by quantitative RT-PCR and western blotting.The differentially regulated proteins (P<0.01) were interrogated against a global molecular network based on the ingenuity knowledge base.The 2D-DIGE analyses revealed DES-induced expression changes for 14 proteins (> 1.3 fold; P<0.05).The iTRAQ analyses allowed the identification of 895proteins.Among these proteins,65 had a modified expression due to DES exposure (i.e.,23 overexpressed and 42 underexpressed).Most of these proteins were implicated in apoptosis and redox processes and had a predicted mitochondrial expression.Additionally,ingenuity pathway analysis placed the OAT and HSBP1 genes at the centre of a highly significant network.RT-PCR confirmed the overexpression of OAT (P=0.006) and HSPB1 (P=0.046).

  7. Xenoestrogens diethylstilbestrol and zearalenone negatively influence pubertal rat's testis.

    Katarzyna Marchlewska


    Full Text Available The aim of this study was to assess the impact of xenoestrogens: diethylstilbestrol (DES and zearalenone (ZEA on rat's pubertal testis and to compare it with the effect of natural estrogen - 17beta-estradiol (E. Male Wistar rats were daily, subcutaneously injected at 5th-15th postnatal days (p.d. with E (1.25 or 12.5 mug or DES (1.25 or 12.5 mug or ZEA (4 or 40 mug or vehicle. At 16th p.d. testes were dissected, weighted, and paraffin embedded. Following parameters were assessed: diameter and length of seminiferous tubule, numbers of spermatogonia A+intermediate+B (A/In/B, preleptotene spermatocytes (PL, leptotene+zygotene+pachytene spermatocytes (L/Z/PA and Sertoli cells per testis. Testes weight, seminiferous tubule diameter and length were decreased by both doses of E, DES and ZEA. DES effect was the strongest, but its influence on testis weight and seminiferous tubule length, on the contrary to E and ZEA, was not dose-dependent. Similarly, DES in both doses had the most severe negative impact on the number of germ and Sertoli cells. The negative influence of E on germ cells was less pronounced. The negative effect of ZEA was seen only after administration of the higher dose on spermatogonia number, while DES and E decreased A/In/B number more evidently. Sertoli cell number were decreased after both doses of E. ZEA40 decreased Sertoli cell number while ZEA4 had no effect. Conclusion: exposure of prepubertal male rat to DES has the strongest detrimental effect on the developing testis in comparison to E and ZEA. Both, E and DES, decreased number of germ and Sertoli cells, diminished seminiferous tubule diameter, length and testis weight. ZEA had much more weaker effect than the potent estrogens.

  8. [Consequences of diethylstilbestrol during pregnancy; 50 years later still a significant problem

    Treffers, P.E.; Hanselaar, A.G.J.M.; Helmerhorst, T.J.M.; Koster, M.E.T.A.; Leeuwen, F.E. van


    Since the 1940s, diethylstilbestrol (DES) has been administered to about three million pregnant women in the United States and in the Netherlands, between 1947 and 1975, to about 220,000. The most important consequences described are: for DES mothers an increased risk of mammary carcinomas and for

  9. Effects of diethylstilbestrol on the proliferation and tyrosinase activity of cultured human melanocytes

    TANG, JIANBING; Li, Qin; Cheng, Biao; HUANG, CHONG; Chen, Kui


    The aim of the present study was to observe the effects of different exogenous estrogen diethylstilbestrol (DES) concentrations on the human melanocyte proliferation and tyrosinase activity. Skin specimens were obtained following blepharoplasty, and the melanocytes were primary cultured and passaged to the third generation. The melanocytes were seeded in 96-well plates, each well had 5×103 cells. The medium was changed after 24 h, and contained 10−4-10−8 M DES. After the melanocytes were incu...

  10. Studies on the mammary tumor-inhibiting effects of diethylstilbestrol and its mono- and diphosphate.

    Schneider, M R; von Angerer, E; Prekajac, J; Brade, W P


    Diethylstilbestrol (DES), diethylstilbestrol monophosphate (DES-MP) and diethylstilbestrol diphosphate (DES-DP) were tested for their estrogen receptor affinity, estrogenic potency and mammary tumor-inhibiting activity in vitro and in vivo. DES had a much higher receptor binding affinity than its mono- or diphosphate. All three compounds inhibited the growth of the hormone-dependent MCF-7 and hormone-independent MDA-MB 231 breast cancer line only at relatively high concentrations. The estrogenic potency in the immature mouse uterine weight test decreased in the order DES greater than DES-MP much greater than DES-DP. The hormone-dependent MXT mammary tumor of the mouse was inhibited by all three compounds at a dosage of 1.0 mg/kg per week. At a dose of 0.01 mg/kg, DES, DES-MP, and DES-DP stimulated the tumor growth. Thus, for the first time, a biphasic effect on tumor growth was demonstrated in intact mature animals. As the effects of all three compounds were similar in this assay, a cleavage of the phosphate groups is likely. A decrease in estrogenic potency concomitant with a retained antitumor effect of DES-MP and DES-DP compared to DES was not demonstrable in the mature mouse using the MXT assay, only in the uterotrophic test in the immature mouse.

  11. Development of an Indirect Competitive ELISA Based on Polyclonal Antibody for the Detection of Diethylstilbestrol in Water Samples

    WANG,Wen-Jun; LING,Yun; XU,Ting; GAO,Hong-Bin; SHENG,Wei; LI,Ji


    An indirect competitive enzyme-linked immunosorbent assay (icELISA) based on polyclonal antibody for the estrogen diethylstilbestrol (DES) was developed. With this aim, two different haptens mono-O-3-carboxypropyldiethylstilbestrol (DES-CP) and mono-O-carboxymethyldiethylstilbestrol (DES-CM) with carboxylic group that preserve the molecular structure character of diethylstilbestrol were synthesized. The haptens were conjugated with the carrier proteins bovine serum albumin (BSA) by mixed-anhydride method for immunogen and conjugated with ovalbumin (OVA) by active ester method for coating antigen. Polyclonal antibodies for diethylstilbestrol were raised by immunizing mice with immune antigen DES-CP-BSA. Under optimized system, the lowest limit of detection (LLD) of diethylstilbestrol was 0.01 ng/mL, and IC50= 1.02 ng/mL. Its analogs were tested and no obvious cross-reactivity was found to anti-diethylstilbestrol antibody. DES-fortified water samples were determined by simple dilution to diminish the matrix effect. The comparison between the amount of DES estimated by ELISA and the amount added indicates good agreement for all water samples tested, with mean recovery values ranging from 86% to 120.2%.

  12. Effects of ginkgo biloba on testicle injury induced by diethylstilbestrol in mice.

    Wang, Wei; Zhong, Xiu-Hui; Ma, Aituan; Shi, Wanyu; Zhang, Xiao-Si; Liu, Yuzhi


    To evaluate the effect of gingko biloba (EGb) on diethylstilbestrol (DES) induced testicle injury in mice. Fifty male mice were divided into a control group (A), DES group (B), and 3 EGb groups (C, D, E). The EGb-treated groups received peritoneal EGb at 8.75 (C), 17.5 (D), 35 mg/kg (E) BW daily for 7 days. The control group was given equivalent amount of normal saline. The mice in groups B, C, D and E were injected hypodermically with DES at 40 mg/kg BW daily 4 hours after the first herbal administration, while the control was given olive oil. Compared with DES group, the testis coefficients-relative testicular weight increased in the three EGb-treated groups. No significant difference was observed in epididymis coefficients. Lipid peroxidation status and antioxidant enzyme activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were significantly elevated in testes of EGb-treated groups. Lactate dehydrogenase (LDH) activities and malonaldehyde (MDA) contents were significantly decreased in testes of the EGb groups. The results indicate that EGb protects the testis from diethylstilbestrol-induced injury.

  13. Exposure to diethylstilbestrol during sensitive life stages: a legacy of heritable health effects.

    Reed, Casey E; Fenton, Suzanne E


    Diethylstilbestrol (DES) is a potent estrogen mimic that was predominantly used from the 1940s to the 1970s by pregnant women in hopes of preventing miscarriage. Decades later, DES is known to enhance breast cancer risk in exposed women and cause a variety of birth-related adverse outcomes in their daughters such as spontaneous abortion, second trimester pregnancy loss, preterm delivery, stillbirth, and neonatal death. Additionally, children exposed to DES in utero suffer from sub/infertility and cancer of reproductive tissues. DES is a pinnacle compound that demonstrates the fetal basis of adult disease. The mechanisms of cancer and endocrine disruption induced by DES are not fully understood. Future studies should focus on common target tissue pathways affected and the health of the DES grandchildren.

  14. μ- and m-calpain expression and activity changes following diethylstilbestrol injection in the rat anterior pituitary

    Weijiang Zhao; Zhongfang Shi; Fang Yuan; Guilin Li; Yazhuo Zhang; Zhongcheng Wang


    Little is known about changes in calpain activity in the pituitary gland.In the present study,μ- and m-calpain activity changes were detected in the rat anterior pituitary following intraperitoneal injection of diethylstilbestrol.Double-immunofluorescence labeling confirmed colocalization of μ - and m-calpain in prolactin-secreting cells (lactotrophs).Western blot analysis revealed significantly increased expression of both calpains,which accompanied upregulated cytosol and membrane zymographic activities at 12 weeks following diethylstilbestrol injection,compared with rats injected with sunflower oil.Moreover,following estrogen injection,pituitary gland pathological damage gradually worsened with increasing time.Results demonstrated that estrogen regulated calpain expression and activity,and both calpains participated in the pathophysiological processes of the pituitary gland.Ubiquitous calpain expression could serve as an effective target for anti-estrogen drugs.

  15. Randomized clinical trial of diethylstilbestrol versus tamoxifen in postmenopausal women with advanced breast cancer.

    Ingle, J N; Ahmann, D L; Green, S J; Edmonson, J H; Bisel, H F; Kvols, L K; Nichols, W C; Creagan, E T; Hahn, R G; Rubin, J; Frytak, S


    Before the introduction of tamoxifen, diethylstilbestrol (DES) was widely considered to be the hormonal treatment of choice in postmenopausal women with advanced breast cancer. We performed a randomized clinical trial of these two agents to determine their relative efficacy and toxicity. The trial involved 143 evaluable patients, of whom 99 had received no prior systemic therapy and 44 had received previous chemotherapy. The regression rates (complete plus partial) were higher in patients receiving DES (41 per cent) than in those receiving tamoxifen (33 per cent), but not significantly so (P = 0.37). In patients who had had no prior systemic therapy, the rates were 44 per cent and 38 per cent, respectively (P = 0.55), and in those who had had previous chemotherapy, 32 per cent vs. 23 per cent (P = 0.50). Analysis of the time until treatment failure for the two treatment groups showed no significant difference (medians: DES, 142 days; tamoxifen, 171 days). Toxicity was greater in patients receiving DES; nine of 74 patients (12 per cent) discontinued therapy solely because of adverse reactions. Since there was no statistically significant difference in efficacy and since tamoxifen was less toxic, tamoxifen appears to be the preferred agent.

  16. Effects of diethylstilbestrol exposure during gestation on both maternal and offspring behavior.

    Kazuya eTomihara


    Full Text Available Endocrine disruption during gestation impairs the physical and behavioral development of offspring. However, it is unclear whether endocrine disruption also impairs maternal behavior and in turn further contributes to the developmental and behavioral dysfunction of offspring. We orally administered the synthetic non-steroidal estrogen diethylstilbestrol (DES to pregnant female C57BL/6J mice from gestation day 11–17 and then investigated the maternal behavior of mothers. In addition, we examined the direct effects of in utero DES exposure and the indirect effects of aberrant maternal behavior on offspring using the cross-fostering method. In mothers, endocrine disruption during gestation decreased maternal behavior. In addition, endocrine disruption of foster mother influenced anxiety-related behavior and passive avoidance learning of pups regardless of their exposure in utero. The influence of DES exposure in utero, irrespective of exposure to the foster mother, was also shown in female offspring. These results demonstrate the risks of endocrine disruptors on both mother as well as offspring and suggest that developmental deficits may stem from both in utero toxicity and aberrant maternal care.

  17. Colestasis intrahepática por dietiletilbestrol Intrahepatic cholestasis due to diethylstilbestrol

    María Julia Cigales Reyes


    Full Text Available El síndrome ictérico es causa frecuente de ingreso en nuestros hospitales, en particular la colestasis, término preferido al de íctero obstructivo, ya que no es imprescindible que exista una obstrucción mecánica. Puede ser provocado por causas intrahepáticas o extrahepáticas, entre las primeras, la toxicidad medicamentosa constituye la segunda causa más frecuente. Se presentó un paciente que se atendió con colestasis intrahepática como efecto adverso del dietiletilbestrol, medicamento que se utiliza frecuentemente para el tratamiento de la neoplasia de próstataThe icteric syndrome is a frequent cause of admission in our hospitals, particularly, cholestasis, a term preferred for obstructive icterus, since the existance of a mechanical obstruction is not indispensable. It may be produced by intrahepatic or extrahepatic causes. Among the first, drug toxicity is the second most common cause. A patient with intrahepatic cholestasis as an adverse effect of diethylstilbestrol, a drug that is usually used for the treatment of prostatic neoplasia, was presented

  18. Neonatal diethylstilbestrol exposure alters the metabolic profile of uterine epithelial cells

    Yan Yin


    Developmental exposure to diethylstilbestrol (DES causes reproductive tract malformations, affects fertility and increases the risk of clear cell carcinoma of the vagina and cervix in humans. Previous studies on a well-established mouse DES model demonstrated that it recapitulates many features of the human syndrome, yet the underlying molecular mechanism is far from clear. Using the neonatal DES mouse model, the present study uses global transcript profiling to systematically explore early gene expression changes in individual epithelial and mesenchymal compartments of the neonatal uterus. Over 900 genes show differential expression upon DES treatment in either one or both tissue layers. Interestingly, multiple components of peroxisome proliferator-activated receptor-γ (PPARγ-mediated adipogenesis and lipid metabolism, including PPARγ itself, are targets of DES in the neonatal uterus. Transmission electron microscopy and Oil-Red O staining further demonstrate a dramatic increase in lipid deposition in uterine epithelial cells upon DES exposure. Neonatal DES exposure also perturbs glucose homeostasis in the uterine epithelium. Some of these neonatal DES-induced metabolic changes appear to last into adulthood, suggesting a permanent effect of DES on energy metabolism in uterine epithelial cells. This study extends the list of biological processes that can be regulated by estrogen or DES, and provides a novel perspective for endocrine disruptor-induced reproductive abnormalities.

  19. Lectin binding patterns in hyperplastic and metaplastic bullock prostate tissues after diethylstilbestrol administration.

    Fernández, P E; Barbeito, C G; Portiansky, E L; Gimeno, E J


    Hyperplasia and squamous metaplasia of the prostatic epithelium are conditions induced by oestrogens. Diethylstilbestrol (DES) has been banned from cattle used for beef production because of the health risks. The potential use of molecular markers for the detection of illegal oestrogen administration was evaluated by taking samples of prostatic tissue from control bullocks, bullocks which had been treated with oestrogens, and bullocks sacrificed 21 and 90 days after a single dose of DES. The expression of the glycoconjugates was examined by lectinhistochemistry and the lectin binding pattern was characterised in epithelium and connective tissue. In the animals sacrificed after 21 days there was an increase in the binding of one lectin (JAC) and there was an increase in the binding of one of the other lectins (DBA) in the animals sacrificed after 90 days. An increase in SWGA lectin staining was observed in the bullocks that had probably been treated with oestrogen and in the animals sacrificed 90 days after the inoculation with DES. There were also differences between the binding of SWGA in the control bullocks and the other groups.

  20. Determination of hepatic uridine 5'-diphosphoglucuronic acid concentration by conjugation with diethylstilbestrol.

    Watkins, J B; Klaassen, C D


    A sensitive and reliable assay for uridine 5'-diphosphoglucuronic acid (UDPGA) was developed that involved conjugation of diethylstilbestrol (DES) in vitro. This conjugation reaction is solely dependent upon UDPGA concentration. The assay uses 0.13 M Tris-HCl, pH 7.4, 6.7 mM MgCl2, 0.05% Brig 58, 0.25 mg guinea pig liver microsomal protein, 0.13 mM 3H-DES (0.2 microCi/ml), and 200 microliters of boiled 10% liver homogenate in a total volume of 0.5 ml. After a 60-min incubation at 37 degrees C, unconjugated DES is extracted into 5 ml of chloroform and the residual metabolized 3H-DES in the aqueous phase is determined by liquid scintillation spectrometry. After addition of beta-glucuronidase to the aqueous phase, about 90% of the radioactivity could be extracted into chloroform, demonstrating the DES-glucuronic acid is the primary metabolite. Thus, this method easily permits quantitation of UDPGA in rat liver in the 1-10 nmol range.

  1. The endocrine disruptor diethylstilbestrol induces adipocyte differentiation and promotes obesity in mice

    Hao, Chan-Juan; Cheng, Xue-Jia; Xia, Hong-Fei, E-mail:; Ma, Xu


    Epidemiology studies indicate that exposure to endocrine disruptors during developmental “window” contributes to adipogenesis and the development of obesity. Implication of endocrine disruptor such as diethylstilbestrol (DES) on adipose tissue development has been poorly investigated. Here we evaluated the effects of DES on adipocyte differentiation in vitro and in vivo, and explored potential mechanism involved in its action. DES induced 3T3-L1 preadipocyte differentiation in a dose-dependent manner, and activated the expression of estrogen receptor (ER) and peroxisome proliferator-acivated receptor (PPAR) γ as well as its target genes required for adipogenesis in vitro. ER mediated the enhancement of DES-induced PPARγ activity. Moreover, DES perturbed key regulators of adipogenesis and lipogenic pathway in vivo. In utero exposure to low dose of DES significantly increased body weight, liver weight and fat mass in female offspring at postnatal day (PND) 60. In addition, serum triglyceride and glucose levels were also significantly elevated. These results suggest that perinatal exposure to DES may be expected to increase the incidence of obesity in a sex-dependent manner and can act as a potential chemical stressor for obesity and obesity-related disorders. -- Highlights: ► DES induced adipocyte differentiation in a dose-dependent manner in 3T3-L1 cells. ► DES activated adipogenic critical regulators and markers in vitro and in vivo. ► Perinatal exposure to DES led to the obese phenotype in female offspring. ► DES might be a potential chemical stressor for obesity and obesity-related disorders.

  2. Urogenital abnormalities in men exposed to diethylstilbestrol in utero: a cohort study

    Palmer Julie R


    Full Text Available Abstract Background Diethylstilbestrol (DES, a synthetic estrogen widely prescribed to pregnant women during the 1940s–70s, has been shown to cause reproductive problems in the daughters. Studies of prenatally-exposed males have yielded conflicting results. Methods In data from a collaborative follow-up of three U.S. cohorts of DES-exposed sons, we examined the relation of prenatal DES exposure to occurrence of male urogenital abnormalities. Exposure status was determined through review of prenatal records. Mailed questionnaires (1994, 1997, 2001 asked about specified abnormalities of the urogenital tract. Risk ratios (RR were estimated by Cox regression with constant time at risk and control for year of birth. Results Prenatal DES exposure was not associated with varicocele, structural abnormalities of the penis, urethral stenosis, benign prostatic hypertrophy, or inflammation/infection of the prostate, urethra, or epididymus. However, RRs were 1.9 (95% confidence interval 1.1–3.4 for cryptorchidism, 2.5 (1.5–4.3 for epididymal cyst, and 2.4 (1.5–4.4 for testicular inflammation/infection. Stronger associations were observed for DES exposure that began before the 11th week of pregnancy: RRs were 2.9 (1.6–5.2 for cryptorchidism, 3.5 (2.0–6.0 for epididymal cyst, and 3.0 (1.7–5.4 for inflammation/infection of testes. Conclusion These results indicate that prenatal exposure to DES increases risk of male urogenital abnormalities and that the association is strongest for exposure that occurs early in gestation. The findings support the hypothesis that endocrine disrupting chemicals may be a cause of the increased prevalence of cryptorchidism that has been seen in recent years.

  3. Diethylstilbestrol in fish tissue determined through subcritical fluid extraction and with GC-MS

    Qiao, Qinghui; Shi, Nianrong; Feng, Xiaomei; Lu, Jie; Han, Yuqian; Xue, Changhu


    As the key point in sex hormone analysis, sample pre-treatment technology has attracted scientists' attention all over the world, and the development trend of sample preparation forwarded to faster and more efficient technologies. Taking economic and environmental concerns into account, subcritical fluid extraction as a faster and more efficient method has stood out as a sample pre-treatment technology. This new extraction technology can overcome the shortcomings of supercritical fluid and achieve higher extraction efficiency at relatively low pressures and temperatures. In this experiment, a simple, sensitive and efficient method has been developed for the determination of diethylstilbestrol (DES) in fish tissue using subcritical 1,1,1,2-tetrafluoroethane (R134a) extraction in combination with gas chromatography-mass spectrometry (GC-MS). After extraction, freezing-lipid filtration was utilized to remove fatty co-extract. Further purification steps were performed with C18 and NH2 solid phase extraction (SPE). Finally, the analyte was derived by heptafluorobutyric anhydride (HFBA), followed by GC-MS analysis. Response surface methodology (RSM) was employed to optimizing the extraction condition, and the optimized was as follows: extraction pressure, 4.3 MPa; extraction temperature, 26°C; amount of co-solvent volume, 4.7 mL. Under this condition, at a spiked level of 1, 5, 10 μg kg-1, the mean recovery of DES was more than 90% with relative standard deviations (RSDs) less than 10%. Finally, the developed method has been successfully used to analyzing the real samples.

  4. Comparison of the disposition of diethylstilbestrol and estradiol in the fetal rat. Correlation with teratogenic potency.

    Henry, E C; Miller, R K


    The dispositions of radiolabeled diethylstilbestrol (DES) and estradiol (E2) in the fetal rat were compared to determine whether kinetic differences accounted for their differences in teratogenic potency. 14C (from DES) was concentrated in fetal tissues relative to plasma, whereas 3H (from E2) was largely retained protein-bound in fetal plasma. Both compounds were rapidly metabolized in the fetus (and mother) to less or non-estrogenic products. Fetal levels of E2 declined faster than those of DES (E1 was the primary circulating estrogen within 1-3 hr of E2 injection) so that exposure to unchanged DES was of longer duration than to E2. The unchanged compounds were retained longer and at higher concentrations in the target genital tissue compared to other tissues. Although these differences were consistent with the potencies, the concentration of the unchanged estrogen in fetal genital tract was lower after a teratogenic dose of DES than after a threshold teratogenic dose of E2. However, the 3H in fetal plasma and genital tract cytosol at 1 hr after injection of [3H]E2 at 2 ng or 10 micrograms/fetus was found to be highly protein-bound. DES competed poorly for these binding sites. It is suggested that the concentration of E2 which is "free" in the cell (as DES is), rather than the total content, correlates with its teratogenicity. Thus, in the rat, rapid metabolism and extensive protein-binding, both extra- and intracellularly, reduce the teratogenicity of the natural estrogen compared to the synthetic estrogen.

  5. Liquid-phase exfoliated graphene as highly-sensitive sensor for simultaneous determination of endocrine disruptors: Diethylstilbestrol and estradiol

    Hu, Lintong; Cheng, Qin [Key Laboratory for Large-Format Battery Materials and System, Ministry of Education, School of Chemistry and Chemical Engineering, Huazhong University of Science and Technology, Wuhan 430074 (China); Chen, Danchao; Ma, Ming [Ningbo Entry-exit Inspection and Quarantine Bureau of China, Ningbo 315012 (China); Wu, Kangbing, E-mail: [Key Laboratory for Large-Format Battery Materials and System, Ministry of Education, School of Chemistry and Chemical Engineering, Huazhong University of Science and Technology, Wuhan 430074 (China)


    Graphical abstract: - Highlights: • A novel electrochemical sensor was developed for diethylstilbestrol and estradiol. • Graphene prepared by solvent exfoliation greatly enhances the detection sensitivity. • The newly-developed method has promising application and the accuracy is good. - Abstract: It is quite important to develop convenient and rapid analytical methods for trace levels of endocrine disruptors because they heavily affect health and reproduction of humans and animals. Herein, graphene was easily prepared via one-step exfoliation using N-methyl-2-pyrrolidone as solvent, and then used to construct an electrochemical sensor for highly-sensitive detection of diethylstilbestrol (DES) and estradiol (E2). On the surface of prepared graphene film, two independent and greatly-increased oxidation waves were observed at 0.28 V and 0.49 V for DES and E2. The remarkable signal enlargements indicated that the detection sensitivity was improved significantly. The influences of pH value, amount of graphene and accumulation time on the oxidation signals of DES and E2 were discussed. As a result, a highly-sensitive and rapid electrochemical method was newly developed for simultaneous detection of DES and E2. The values of detection limit were evaluated to be 10.87 nM and 4.9 nM for DES and E2. Additionally, this new method was successfully used in lake water samples and the accuracy was satisfactory.

  6. Liver X receptors interfere with the deleterious effect of diethylstilbestrol on testicular physiology

    Oumeddour, Abdelkader [Clermont Université, Université Blaise Pascal, Génétique Reproduction et Développement, BP 10448, F-63000 Clermont-Ferrand (France); CNRS, UMR 6293, GReD, F-63171 Aubiere (France); INSERM, UMR 1103, GReD, F-63171 Aubiere (France); Centre de Recherche en Nutrition Humaine d’Auvergne, F-63000 Clermont-Ferrand (France); Laboratoire de Neuroendocrinologie Appliquée, Université Badji Mokhtar Annaba, BP12, 23000 Annaba (Algeria); Viennois, Emilie; Caira, Françoise; Decourbey, Clélia [Clermont Université, Université Blaise Pascal, Génétique Reproduction et Développement, BP 10448, F-63000 Clermont-Ferrand (France); CNRS, UMR 6293, GReD, F-63171 Aubiere (France); INSERM, UMR 1103, GReD, F-63171 Aubiere (France); Centre de Recherche en Nutrition Humaine d’Auvergne, F-63000 Clermont-Ferrand (France); Maqdasy, Salwan [Clermont Université, Université Blaise Pascal, Génétique Reproduction et Développement, BP 10448, F-63000 Clermont-Ferrand (France); CNRS, UMR 6293, GReD, F-63171 Aubiere (France); INSERM, UMR 1103, GReD, F-63171 Aubiere (France); Centre de Recherche en Nutrition Humaine d’Auvergne, F-63000 Clermont-Ferrand (France); Service d’endocrinologie, diabétologie et maladies métaboliques, CHU Clermont-Ferrand, F-63003 Clermont-Ferrand (France); and others


    Highlights: • Part of the neonatal effect of DES on testis needs the presence of Lxrα/β. • Some DES-induced pathways are blocked in Lxr-deficient mice. • Lxr-deficient mice analysis defines DES-target genes protected by Lxr. - Abstract: Liver X receptors LXRα (NR1H3) and LXRβ (NR1H2) are transcription factors belonging to the nuclear receptor superfamily, activated by specific oxysterols, oxidized derivatives of cholesterol. These receptors are involved in the regulation of testis physiology. Lxr-deficient mice pointed to the physiological roles of these nuclear receptors in steroid synthesis, lipid homeostasis and germ cell apoptosis and proliferation. Diethylstilbestrol (DES) is a synthetic estrogen considered as an endocrine disruptor that affects the functions of the testis. Various lines of evidences have made a clear link between estrogens, their nuclear receptors ERα (NR3A1) and ERβ (NR3A2), and Lxrα/β. As LXR activity could also be regulated by the nuclear receptor small heterodimer partner (SHP, NR0A2) and DES could act through SHP, we wondered whether LXR could be targeted by estrogen-like endocrine disruptors such as DES. For that purpose, wild-type and Lxr-deficient mice were daily treated with 0.75 μg DES from days 1 to 5 after birth. The effects of DES were investigated at 10 or 45 days of age. We demonstrated that DES induced a decrease of the body mass at 10 days only in the Lxr-deficient mice suggesting a protective effect of Lxr. We defined three categories of DES-target genes in testis: those whose accumulation is independent of Lxr; those whose accumulation is enhanced by the lack of both Lxrα/β; those whose accumulation is repressed by the absence of Lxrα/β. Lipid accumulation is also modified by neonatal DES injection. Lxr-deficient mice present different lipid profiles, demonstrating that DES could have its effects in part due to Lxrα/β. Altogether, our study shows that both nuclear receptors Lxrα and Lxrβ are not only

  7. Comparative activities of p-nonylphenol and diethylstilbestrol in noble rat mammary gland and uterotrophic assays.

    Odum, J; Pyrah, I T; Foster, J R; Van Miller, J P; Joiner, R L; Ashby, J


    Colerangle and Roy (1996, Endocrine 4, 115-122) have described the apparent ability of both diethylstilbestrol (DES) and p-nonylphenol (NP) to cause extensive cell proliferation and lobular development in the mammary glands of young adult Noble rats. The chemicals were administered over 11 days via subcutaneously implanted minipumps. The dose level of DES used (0.076 mg/kg/day) was about 70 times higher than its minimum detection level in rodent uterotrophic and reproductive toxicology studies. In contrast, the lowest active dose level of NP (0.073 mg/kg/day) in the Noble rat mammary gland study was about 600 times lower than its minimum detection level in rat uterotrophic and multigeneration studies. The apparent enhanced sensitivity of the Noble rat mammary gland to the estrogenic activity of NP was considered worthy of further study. Ovariectomized Noble rat uterotrophic assays with NP (minimum detection level approximately 40 mg/kg/day, 3 or 11 days, oral gavage) revealed similar assay sensitivity to that observed for earlier immature and ovariectomized Alderley Park (AP) rat uterotrophic assays of this chemical. The response of the ovariectomized Noble rat uterotrophic assay to DES and estradiol was also as expected from earlier immature AP rat assays. It is concluded that the general sensitivity to estrogens of the Noble rat and the AP rat is similar. A repeat of the Noble rat mammary gland study with DES (11 x 0.076 mg/kg/day) and NP (11 x either 0.073 or 53.2 mg/kg/day), as originally reported by Colerangle and Roy (1996), revealed a strong positive response to DES and no response to NP. It is concluded that the minimum detection level of NP as a weakly estrogenic material in the rat should be based on the results of rat uterotrophic and multigeneration studies and therefore be set at approximately 40 mg/kg/day. It is also concluded that induced S-phase in the rodent mammary gland is best monitored using BRDU, as opposed to PCNA staining, and that use of

  8. Diethylstilbestrol-diphosphate induces chromosomal aberrations but not sister chromatid exchanges in murine bone marrow cells in vivo

    Ivett, J.L. (North Carolina State Univ., Raleigh); Tice, R.R.


    Diethylstilbestrol diphosphate (DES-dp) clastogenesis was examined in the bone marrow of C57B1/6 male and female mice. Significant and sex-related dose effects were observed for the induction of chromatid-type chromosomal aberrations and for the inhibition of cellular proliferation. Females were more sensitive to the effects of DES-dp than males when assessed for either induced chromosomal aberrations or proliferative inhibition. Contrary to other published results, we did not observe either an increase in sister chromatid exchanges or an increased incidence of aneuploidy. Ovariectomy reduced the ability of DES-dp to inhibit cellular proliferation and decreased the high degree of variability between animals at high doses of DES-dp. The results of our studies show that DES is a clastogenic agent in vivo which may relate to its carcinogenicity.

  9. Liquid-phase exfoliated graphene as highly-sensitive sensor for simultaneous determination of endocrine disruptors: diethylstilbestrol and estradiol.

    Hu, Lintong; Cheng, Qin; Chen, Danchao; Ma, Ming; Wu, Kangbing


    It is quite important to develop convenient and rapid analytical methods for trace levels of endocrine disruptors because they heavily affect health and reproduction of humans and animals. Herein, graphene was easily prepared via one-step exfoliation using N-methyl-2-pyrrolidone as solvent, and then used to construct an electrochemical sensor for highly-sensitive detection of diethylstilbestrol (DES) and estradiol (E2). On the surface of prepared graphene film, two independent and greatly-increased oxidation waves were observed at 0.28V and 0.49V for DES and E2. The remarkable signal enlargements indicated that the detection sensitivity was improved significantly. The influences of pH value, amount of graphene and accumulation time on the oxidation signals of DES and E2 were discussed. As a result, a highly-sensitive and rapid electrochemical method was newly developed for simultaneous detection of DES and E2. The values of detection limit were evaluated to be 10.87 nM and 4.9 nM for DES and E2. Additionally, this new method was successfully used in lake water samples and the accuracy was satisfactory.

  10. Persulfate-assisted photodegradation of diethylstilbestrol using monoclinic BiVO4 under visible-light irradiation.

    Liu, Yang; Zhang, Yongli; Guo, Hongguang; Cheng, Xin; Liu, Hongwei; Tang, Weihong


    In this study, the photosynergistic performance of BiVO4 with persulfate (PS) is demonstrated under visible light irradiation for the first time. Diethylstilbestrol (DES) was selected as a reluctant compound, and factors including dosages of PS and catalyst, solution pHs, initial concertration of DES, and inorganic anions were evaluated. The morphology and chemical state of bismuth vanadate (BiVO4) was characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive spectrometer (EDS), and ultraviolet-visible (UV-Vis) diffuse reflectance spectroscopy (DRS). It was found that the degradation of DES was promoted in either acid or alkaline solutions. The increase of PS and BiVO4 dosages was beneficial to the reactions, while incremental concentration of DES showed the inhibiting effect. By scavenging hVB(+), Cl(-) was able to make the promotion, differentiated from the exsiting HCO- 3. Moreover, the photocatalytic mechanism for the BiVO4/PS/vis-light system was proposed by using several probe compounds (isopropanol, tert-butanol, and 1,4-benzoquinone), which consists of h+ VB/e- CB generation and recombination on the surface of BiVO4 as well as free radical oxidation in the solutions. The study provides a distinctive method to treat organic contaminants using visible light in the aqueous environment.

  11. Histone methyltransferase EZH2 is transcriptionally induced by estradiol as well as estrogenic endocrine disruptors bisphenol-A and diethylstilbestrol.

    Bhan, Arunoday; Hussain, Imran; Ansari, Khairul I; Bobzean, Samara A M; Perrotti, Linda I; Mandal, Subhrangsu S


    Enhancer of Zeste homolog 2 (EZH2), a methyltransferase specific to histone 3 lysine 27, is a critical player in gene silencing and is overexpressed in breast cancer. Our studies demonstrate that EZH2 is transcriptionally induced by estradiol in cultured breast cancer cells and in the mammary glands of ovariectomized rats. EZH2 promoter contains multiple functional estrogen-response elements. Estrogen receptors (ERs) and ER coregulators such as mixed lineage leukemia (MLL) histone methylases (MLL2 and MLL3) and histone acetyltransferase CBP/P300 bind to the EZH2 promoter in the presence of estradiol and regulate estradiol-induced EZH2 expression. EZH2 expression is also increased upon exposure to estrogenic endocrine disrupting chemicals (EDCs) such as bisphenol-A (BPA) and diethylstilbestrol (DES). Similar to estradiol, BPA and DES-induced EZH2 expression is coordinated by ERs, MLLs and CBP/P300. In summary, we demonstrate that EZH2 is transcriptionally regulated by estradiol in vitro and in vivo, and its expression is potentially dysregulated upon exposure to estrogenic EDCs.

  12. Expression of c-fos and c-jun protooncogenes in the uteri of immature mice neonatally exposed to diethylstilbestrol.

    Yamashita, S; Takayanagi, A; Shimizu, N


    We studied the cell-type-specific and temporal expression of c-fos and c-jun protooncogenes after 17beta-estradiol (E2) stimulation in the uteri of immature 3-week-old mice neonatally exposed to diethylstilbestrol (DES), DES-mice, and the ontogenic expression of these genes in the uteri of DES-mice using immunohistochemistry and in situ hybridization. A single E2 injection induced the transient and rapid expression of c-fos mRNA and c-Fos protein in the endometrial epithelium and endothelial cells of the blood vessels in both 3-week-old vehicle-treated controls and DES-mice; a peak of mRNA expression was 2 hours after E2 injection and that of protein expression was 2 to 3 hours after the injection. The expression of c-fos mRNA and protein after E2 stimulation was lower in the DES-mice than in the control animals. There were no significant differences in the c-jun expression patterns in both experimental groups before and after the E2 injection. The E2 injection transiently down-regulated the c-jun expression in the epithelium and up-regulated it in the stroma and myometrium. The uterine epithelium of DES-mice showed much stronger c-Jun immunostaining on days 4 and 10, compared with those of controls. Neonatal DES treatment reduced c-Jun immunoreactivity in the uterine epithelium on days 4 and 10, and increased the reaction in the stroma on day 4. These results suggested that the neonatal DES treatment induces permanent changes in the c-fos expression pattern independent of the postpuberal secretion of ovarian steroids. The changes in the expression of c-fos and c-jun protooncogenes, particularly during postnatal development, are likely to play important roles in the production of uterine abnormalities in the DES-mice.

  13. Expression of Sex-Related Genes in Chicken Embryos During Male-to-Female Sex Reversal Exposure to Diethylstilbestrol

    FANG Li-xiu; XIN Rui; CHE Yi; XU Shi-qing


    Sex emerges out of a delicate dance between a variety of promale, anti-male, and possibly profemale genes. To investigate the role that sex-related genes play in sex determination and gonadal differentiation of fowl, we constructed a male-to-female sex-reversal model of chick induced by diethylstilbestrol (DES) at onset of incubation (E0). The results of semi-quantitative PCR showed that the expression of Sf1, the orphan nuclear receptor steroidogenic factor-1 gene, was put forward from E7d to E5d and up-regulated during E5-7d;the Dmrt1, the double sex and the Mab-3 related to transcription factor 1 gene, was down-regulated during E3-7d. Meanwhile, anti-Müllerian hormone gene (Amh) expressed at a similar level in the genetic females and sex-reversal females before E7d, while no expression products of the three female-specific genes Wpkci, Fet1 and Foxl2 were detected in male-to-female embryos. These findings suggest that the expression of some certain sex-related genes, induced by the exogenous estrogen during period of sex determination and gonadal differentiation, results in the male-to-female sex reversal. Moreover, high activity of Sf1 gene during E5-7d might be related to the profemale process, while low activity of Dmrt1 gene during E3-5d might be anti-male. The expression activity of Amh gene might only contribute to the promale process after E7d, however, it is possibly not an anti-female gene in chick embryos.

  14. The nonsteroidal effects of diethylstilbestrol: the rationale for androgen deprivation therapy without estrogen deprivation in the treatment of prostate cancer.

    Scherr, Douglas S; Pitts, W Reid


    During the last 2 decades there has been an increase in the number of men with prostate cancer placed on luteinizing hormone releasing hormone (LH-RH) agonist therapy. In addition, the duration of individual therapy has extended from what was once only a few months to, in many cases, several years. As a result there has been an increase in the incidence of side effects, including osteoporosis, decreased cognitive abilities, vascular stiffness and fatigue. We explored the use of estrogen in the form of diethylstilbestrol (DES) as an alternative treatment for men with prostate cancer, and introduce the concept of androgen deprivation without estrogen deprivation. In doing so we hope to elucidate some of the nonhormonal nonsteroidal effects of DES. Furthermore, we hope to define the mechanisms by which DES can be useful when LH-RH agonist therapy or orchiectomy has failed. We comprehensively reviewed the literature from 1935 to the present regarding estrogen and antiandrogen therapy. Our search focused on issues pertaining to side effects, efficacy and nonsteroidal effects of antiandrogens and estrogens. It is readily apparent from the literature that androgen deprivation with DES can achieve effective prostate cancer control with demonstrable benefits compared to conventional LH-RH agonist therapy. In particular, rates of bone resorption and osteoporosis are less with the use of estrogen therapies. Estrogen has a clear beneficial effect on cognitive function. The estrogen metabolite 2-methoxyestradiol has significant antiangiogenic and pro-apoptotic effects. These effects give estrogens an added anticancer effect not otherwise seen in conventional LH-RH agonist therapy. The efficacy of 1 mg DES extends well beyond its androgen suppressive effects. Androgen deprivation without estrogen deprivation is a concept that deserves further attention in the urological community.

  15. Gestational exposure to diethylstilbestrol alters cardiac structure/function, protein expression and DNA methylation in adult male mice progeny

    Haddad, Rami, E-mail: [Lady Davis Institute for Medical Research, Jewish General Hospital, 3755 chemin Cote Ste Catherine, Montréal, Québec, Canada H3T 1E2 (Canada); Division of Experimental Medicine, Department of Medicine, McGill University, 850 Sherbrooke Street, Montréal, Québec, Canada H3A 1A2 (Canada); Kasneci, Amanda, E-mail: [Lady Davis Institute for Medical Research, Jewish General Hospital, 3755 chemin Cote Ste Catherine, Montréal, Québec, Canada H3T 1E2 (Canada); Mepham, Kathryn, E-mail: [Lady Davis Institute for Medical Research, Jewish General Hospital, 3755 chemin Cote Ste Catherine, Montréal, Québec, Canada H3T 1E2 (Canada); Division of Experimental Medicine, Department of Medicine, McGill University, 850 Sherbrooke Street, Montréal, Québec, Canada H3A 1A2 (Canada); Sebag, Igal A., E-mail: [Division of Cardiology, Jewish General Hospital, 3755 chemin Cote Ste Catherine, Montréal, Québec, Canada H3T 1E2 (Canada); and others


    Pregnant women, and thus their fetuses, are exposed to many endocrine disruptor compounds (EDCs). Fetal cardiomyocytes express sex hormone receptors making them potentially susceptible to re-programming by estrogenizing EDCs. Diethylstilbestrol (DES) is a proto-typical, non-steroidal estrogen. We hypothesized that changes in adult cardiac structure/function after gestational exposure to the test compound DES would be a proof in principle for the possibility of estrogenizing environmental EDCs to also alter the fetal heart. Vehicle (peanut oil) or DES (0.1, 1.0 and 10.0 μg/kg/da.) was orally delivered to pregnant C57bl/6n dams on gestation days 11.5–14.5. At 3 months, male progeny were left sedentary or were swim trained for 4 weeks. Echocardiography of isoflurane anesthetized mice revealed similar cardiac structure/function in all sedentary mice, but evidence of systolic dysfunction and increased diastolic relaxation after swim training at higher DES doses. The calcium homeostasis proteins, SERCA2a, phospholamban, phospho-serine 16 phospholamban and calsequestrin 2, are important for cardiac contraction and relaxation. Immunoblot analyses of ventricle homogenates showed increased expression of SERCA2a and calsequestrin 2 in DES mice and greater molecular remodeling of these proteins and phospho-serine 16 phospholamban in swim trained DES mice. DES increased cardiac DNA methyltransferase 3a expression and DNA methylation in the CpG island within the calsequestrin 2 promoter in heart. Thus, gestational DES epigenetically altered ventricular DNA, altered cardiac function and expression, and reduced the ability of adult progeny to cardiac remodel when physically challenged. We conclude that gestational exposure to estrogenizing EDCs may impact cardiac structure/function in adult males. -- Highlights: ► Gestational DES changes cardiac SERCA2a and CASQ2 expression. ► Echocardiography identified systolic dysfunction and increased diastolic relaxation. ► DES

  16. Xeno-oestrogens Bisphenol A and Diethylstilbestrol Selectively Activating Androgen Receptor Mediated AREs-TATA Reporter System

    WU Jing; WEI Wei; YANG Nan-yang; SHEN Xiao-yan; TSUJI Ichiro; YAMAMURA Takaki; LI Jiang


    We cloned the three androgen response elements(AREs,including AREⅠ,AREⅡ,and AREⅢ) with a core transactivation TATA element of the prostate-specific antigen(PSA) promoter into pGL2 basic vector to create an artificial pGL2/AREs-TATA reporter system,which was applied to evaluating the effects of different xenooestrogens[bisphenol A(BPA),4-nonylphenol(4-NP),dichlorodiphenyl trichloroethane(DDT) or diethylstilbestrol (DES)] on androgen receptor(AR) abnormal activation to regulate PSA expression and cell proliferation.In all the three AREs,AREⅢ-TATA displayed as a major element responsive to AR-mediated DHT stimulation of PSA promoter.Therefore,pGL2/AREⅢ-TATA reporter was adopted to analyze the activation capacity of AR activated by four different xeno-oestrogens.The activation ofpGL2/AREⅢ-TATA reporter by each xeno-oestrogen was analyzed in two different cell lines,one was HEK293T(Human Embryonic Kidney 293T) cell line,and the other was AR stably expressed DU145 cell line,which was produced by infecting AR with pLenti-puro-AR into the prostate cancer DU145 cells and that were scanned with puromycin and tested by AR antibody.In both the two cell lines,BPA or DES significantly induced AR-mediated transcriptional activity of AREⅢ-TATA reporter,whereas DDT or 4-nonylphenol did not.Moreover,AR-mediated cell proliferation in response to each of four xeno-oestrogens was measured in MTT assays in both HEK293T cell or AR stably expressed DU145 cell lines.BPA or DES,as an AR inducer,exhibited an enhanced effect in cell proliferation,rather than the effect of DDT or 4-NP,in both cell lines.Finally,we demonstrated that BPA or DES stimulated PSA expression and enhanced the recruitment of AR onto thePSA promoter,resulting in stronger binding to AREⅢ sites.Taken together,four xeno-oestrogens were identified to have different activities on AR.BPA and DES are demonstrated to be androgenic effectors in the regulation of PSA activation or cell proliferation.

  17. Subchronic exposure to phytoestrogens alone and in combination with diethylstilbestrol - pituitary tumor induction in Fischer 344 rats

    Kaphalia Bhupendra S


    Full Text Available Abstract Background Subchronic administration of the potent pharmaceutical estrogen diethylstilbestrol (DES to female Fischer 344 (F344 rats induces growth of large, hemorrhagic pituitaries that progress to tumors. Phytoestrogens (dietary plant estrogens are hypothesized to be potential tumor inhibitors in tissues prone to estrogen-induced cancers, and have been suggested as "safer" estrogen replacements. However, it is unknown if they might themselves establish or exacerbate the growth of estrogen-responsive cancers, such as in pituitary. Methods We implanted rats with silastic capsules containing 5 mg of four different phytoestrogens - either coumestrol, daidzein, genistein, or trans-resveratrol, in the presence or absence of DES. We examined pituitary and other organ weights, blood levels of prolactin (PRL and growth hormone (GH, body weights, and pituitary tissue histology. Results Blood level measurements of the administered phytoestrogens confirmed successful exposure of the animals to high levels of these compounds. By themselves, no phytoestrogen increased pituitary weights or serum PRL levels after 10 weeks of treatment. DES, genistein, and resveratrol increased GH levels during this time. Phytoestrogens neither changed any wet organ weight (uterus, ovary, cervix, liver, and kidney after 10 weeks of treatment, nor reversed the adverse effects of DES on pituitaries, GH and PRL levels, or body weight gain after 8 weeks of co-treatment. However, they did reverse the DES-induced weight increase on the ovary and cervix. Morphometric examination of pituitaries revealed that treatment with DES, either alone or in combination with phytoestrogens, caused gross structural changes that included decreases in tissue cell density, increases in vascularity, and multiple hemorrhagic areas. DES, especially in combination with phytoestrogens, caused the development of larger and more heterogeneous nuclear sizes in pituitary. Conclusions High levels of

  18. Diethylstilbestrol (DES) and Cancer

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  19. Diethylstilbestrol can effectively accelerate estradiol-17-O-glucuronidation, while potently inhibiting estradiol-3-O-glucuronidation

    Zhu, Liangliang; Xiao, Ling [The Centre for Drug and Food Safety Evaluation, School of Life Science, Anqing Normal University, Anqing 246011 (China); Xia, Yangliu [Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023 (China); Zhou, Kun [College of Pharmacy, Liaoning University of Traditional Chinese Medicine, Dalian 116600 (China); Wang, Huili; Huang, Minyi [The Centre for Drug and Food Safety Evaluation, School of Life Science, Anqing Normal University, Anqing 246011 (China); Ge, Guangbo, E-mail: [Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023 (China); Wu, Yan; Wu, Ganlin [The Centre for Drug and Food Safety Evaluation, School of Life Science, Anqing Normal University, Anqing 246011 (China); Yang, Ling, E-mail: [Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023 (China)


    This in vitro study investigates the effects of diethylstilbestrol (DES), a widely used toxic synthetic estrogen, on estradiol-3- and 17-O- (E2-3/17-O) glucuronidation, via culturing human liver microsomes (HLMs) or recombinant UDP-glucuronosyltransferases (UGTs) with DES and E2. DES can potently inhibit E2-3-O-glucuronidation in HLM, a probe reaction for UGT1A1. Kinetic assays indicate that the inhibition follows a competitive inhibition mechanism, with the Ki value of 2.1 ± 0.3 μM, which is less than the possible in vivo level. In contrast to the inhibition on E2-3-O-glucuronidation, the acceleration is observed on E2-17-O-glucuronidation in HLM, in which cholestatic E2-17-O-glucuronide is generated. In the presence of DES (0–6.25 μM), K{sub m} values for E2-17-O-glucuronidation are located in the range of 7.2–7.4 μM, while V{sub max} values range from 0.38 to 1.54 nmol/min/mg. The mechanism behind the activation in HLM is further demonstrated by the fact that DES can efficiently elevate the activity of UGT1A4 in catalyzing E2-17-O-glucuronidation. The presence of DES (2 μM) can elevate V{sub max} from 0.016 to 0.81 nmol/min/mg, while lifting K{sub m} in a much lesser extent from 4.4 to 11 μM. Activation of E2-17-O-glucuronidation is well described by a two binding site model, with K{sub A}, α, and β values of 0.077 ± 0.18 μM, 3.3 ± 1.1 and 104 ± 56, respectively. However, diverse effects of DES towards E2-3/17-O-glucuronidation are not observed in liver microsomes from several common experimental animals. In summary, this study issues new potential toxic mechanisms for DES: potently inhibiting the activity of UGT1A1 and powerfully accelerating the formation of cholestatic E2-17-O-glucuronide by UGT1A4. - Highlights: • E2-3-O-glucuronidation in HLM is inhibited when co-incubated with DES. • E2-17-O-glucuronidation in HLM is stimulated when co-incubated with DES. • Acceleration of E2-17-O-glucuronidationin in HLM by DES is via activating the

  20. 高效液相色谱内标法测定鸡肉中己烯雌酚残留量%Determination of Diethylstilbestrol in Chicken Meat by HPLC Internal Standard Method

    龙朝阳; 吴西梅; 朱炳辉; 邹小勇


      采用高效液相色谱法以对羟基苯甲酸丁酯作为内标定量测定了鸡肉中己烯雌酚的含量.色谱柱为C18柱,0.02 mol/L三乙胺水溶液(加磷酸调pH=3.0)-乙腈(v/v=55/45)作为流动相,采用等度洗脱,检测波长为240 nm.结果显示,己烯雌酚的线性范围为0.324~1.30µg/mL,相关系数为0.9998,用浓度为0.648µg/mL的标准溶液重复进样6次,其相对标准偏差(RSD)为1.28%.方法简便准确,适于鸡肉组织中违禁药物己烯雌酚含量的测定.%  The HPLC method was established for the determination of diethylstilbestrol in chicken meat. Diethylstilbestrol and internal standard butyl paraben were separated on a C18 column and detected at 240nm. The mobile phase consisted of 40%actonitrile and 0.02 mol/L trimethylamine (adjust pH to 3.0 by phosphoric acid). Result of the standard curve method is accorded with that of internal standard method. The concentration of diethylstilbestrol was calculated by the internal standard method. The linearity between peak area and concentration of diethylstilbestrol existed in the range of 0.324~1.30 µg/mL and RSD of peak area was in the level of 1.28%. The recovery of diethylstilbestrol was from 98.8%~101.2%(n=3). The method is stable, practical and suitable for the quantitative determination of diethylstilbestrol in chicken meat.

  1. Binding and biologic activity of diethylstilbestrol in the hamster: influence of a serum component on estrogen receptor binding and estrogenic activity.

    Okulicz, W C; Leavitt, W W


    The purpose of this study was to compare the biological activity and estrogen receptor (Re) binding affinity of diethylstilbestrol (DES) and estradiol (E2). Uterine weight response and cytosolic progesterone receptor (Rp) induction were equivalent following daily (3 days) injections of DES or E2 to ovariectomized animals. The biological equivalence of DES and E2 was not reflected by competition assays done with either uterine cytosolic or nuclear Re: the relative binding affinity (RBA) of DES to cytosolic Re was 46 +/- 5.3 and to nuclear Re was 380 +/- 42 compared to E2 (100). The RBAs of estrone, estriol and enclomiphene with cytosolic or nuclear Re were not significantly different. Further studies showed that this discrepancy in RBA of DES between cytosolic and nuclear Re could not be attributed to salt concentration but could be mimicked by addition of serum to nuclear Re preparations. The RBA of DES done with ammonium sulfate precipitated cytosolic Re approached that observed for nuclear Re. Gel filtration chromatography (Sephacryl S-300) of serum bound tritiated DES was shown to coelute with bovine serum albumin. These results suggest that a serum component (tentatively identified as albumin) can bind DES and cause a decrease in in vitro binding affinity and a reduction in biological activity in vivo.

  2. Diethylstilbestrol Exposure in Neonatal Mice Induces Changes in the Adulthood in the Immune Response to Taenia crassiceps without Modifications of Parasite Loads

    Karen E. Nava-Castro


    Full Text Available Industrial growth has increased the exposition to endocrine disruptor compounds (EDC’s, which are exogenous agents with agonist or antagonist action of endogenous steroid hormones that may affect the course of parasite infections. We wanted to determine if the exposure to diethylstilbestrol (DES, an estrogen agonist, to both male and female mice affected the immune response and their susceptibility to T. crassiceps cysticercosis. In all infected groups, females showed higher parasite loads than males, and neonatal DES administration did not modify this pattern. In the spleen, noninfected mice showed sex-related differences in the percentage of the CD8+ subpopulation, but DES decreased the percentage of CD3+, CD19+, and CD8+ subpopulations in infected mice. In the mesenteric lymphatic node (MNL, DES showed a dimorphic effect in the percentage of CD19+ cells. Regarding estrogen receptor alpha (ER-α expression, DES treatment induced a reduction in the expression of this receptor in both noninfected female and male mice in the spleen, which was decreased only in males in CD3+ and CD8+ lymphocytes in MNL cell subpopulations. Our study is the first one to demonstrate that DES neonatal treatment in male and female mice affects the immune cell percentage, without effect on the susceptibility to T. crassiceps cysticercosis.

  3. Involvement of gonadotropins in the induction of hypertrophy-hyperplasia in the interstitial tissues of ovaries in neonatally diethylstilbestrol-treated mice.

    Kakuta, Hanako; Tanaka, Masami; Chambon, Pierre; Watanabe, Hajime; Iguchi, Taisen; Sato, Tomomi


    Neonatally diethylstilbestrol (DES) treatment causes hypertrophy-hyperplasia in the interstitial tissue of mouse ovaries. To understand the induction mechanism of the hypertrophy, mRNA expression involved in steroidogenesis in the ovary of neonatally DES-treated mice was examined. The expression of StAR and Cyp11a1 was significantly reduced while Cyp19 and Sf-1 were stimulated in the ovary of neonatally DES-treated 3-month-old mice. Expression of those genes was not different between DES- and oil-treated mice after the gonadotropins treatment. Lhb in the pituitary of 3-month-old neonatally DES-treated mice was significantly decreased. Finally, ovaries from DES-treated mice transplanted to neonatally oil-treated hosts had developing follicles at several stages and corpora lutea, whereas grafted ovaries from neonatally oil-treated mice in 3-month-old neonatally DES-treated hosts showed lipid accumulation in the interstitial tissue. Thus, hypertrophy and accumulation of lipid droplets in interstitial cells of neonatally DES-treated mice is caused by impaired steroidogenesis due to the alterations of gonadotropins levels.

  4. Modulation of adult rat benzo(a)pyrene (BaP) metabolism and DNA adduct formation by neonatal diethylstilbestrol (DES) exposure.

    Ramesh, Aramandla; Inyang, Frank; Knuckles, Maurice E


    This study seeks to elucidate the role of diethylstilbestrol (DES), a synthetic estrogen on benzo(a)pyrene (BaP) metabolism in the male rat reproductive tissues. Offspring of timed-pregnant Sprague-Dawley rats were neonatally treated on days 2, 4, and 6 post-partum with 1.45 micromol/kg of DES. Ten weeks after birth, the adult rats were challenged with radiolabeled benzo(a)pyrene (3H BaP) (10 micromol/kg) and the rats were sacrificed 2 h after BaP exposure. Prostrate, testis, lung, liver, urine and feces samples were collected and extracted using a mixture of H2O, MeOH and CHCl3. The extracts were analyzed by reverse phase HPLC. The concentrations of BaP organic metabolites in DES rats were lower compared to controls (vehicle-treated rats). On the other hand, concentrations of aqueous metabolites were significantly increased in DES treated animals. The toxication to detoxication ratios were significantly decreased in DES rats compared to controls. This trend is also reflected in the decreased concentrations of BaP-DNA adducts in DES rats. Collectively these results suggest that DES is capable of modulating the metabolic pathway of BaP towards detoxification thereby preventing the manifestation of toxicity.

  5. Brief maternal exposure of rats to the xenobiotics dibutyl phthalate or diethylstilbestrol alters adult-type Leydig cell development in male offspring

    Richard Ivell; Kee Heng; Helen Nicholson; Ravinder Anand-Ivell


    Maternal exposure to estrogenic xenobiotics or phthalates has been implicated in the distortion of early male reproductive development,referred to in humans as the testicular dysgenesis syndrome.It is not known,however,whether such early gestational and/or lactational exposure can influence the later adult-type Leydig cell phenotype.In this study,Sprague-Dawley rats were exposed to dibutyl phthalate (DBP; from gestational day (GD) 14.5 to postnatal day (PND) 6) or diethylstilbestrol (DES; from GD14.5 to GD16.5) during a short gestational/lactational window,and male offspring subsequently analysed for various postnatal testicular parameters.All offspring remained in good health throughout the study.Maternal xenobiotic treatment appeared to modify specific Leydig cell gene expression in male offspring,particularly during the dynamic phase of mid-puberty,with serum INSL3 concentrations showing that these compounds led to a faster attainment of peak values,and a modest acceleration of the pubertal trajectory.Part of this effect appeared to be due to a treatment-specific impact on Leydig cell proliferation during puberty for both xenobiotics.Taken together,these results support the notion that maternal exposure to certain xenobiotics can also influence the development of the adult-type Leydig cell population,possibly through an effect on the Leydig stem cell population.

  6. 含己烯雌酚血清对体外培养正常人黑素细胞的作用%Effects of Serum containing Diethylstilbestrol (DES)on cultured human melanocytes in vitro

    龚石; 杨先旭; 张晴; 刘巧


    Objective Establishing the culture of human normal melanocytes in vitro,and studying the effects of Serum containing Diethylstilbestrol(DES)on cultured human melanocytes. Methods Viability of melanocytes were measured by MTT method; tyrosinase activity was measured by utilization of L-Dopa as the substrate (Nakajima M method ) and melanocytes melanogenesi by NaOH method. Results Different dosage of Serum containing Diethylstilbestroi (DES) induced a strong promotion on melanocytic viability and tyrosinase activity. Conclusion Serum containing Diethylstilbestrol (DES) can promote melanocytic proliferation and tyrosinase activity. Serum containing Diethylstilbestrol might have some therapenticai effects on correlated pigmented skin diseases .%目的:观察不同浓度含己烯雌酚(DES)血清对体外培养人黑素细胞的增殖和酪氨酸酶活性的影响.方法:采用MTT法测定黑素细胞增殖情况;参考Nakajima M方法测定酪氨酸酶活性;NaOH溶解法测定黑素含量.结果:己烯雌酚血清不同剂量对人黑素细胞增殖及酪氨酸酶活性均有促进作用.结论:己烯雌酚能促进体外培养人黑素细胞增殖及酪氨酸酶活性,对相关皮肤色素性疾病可能有一定的治疗作用.

  7. FSH and LH Secretion from in-vitro Cultured Buffalo Anterior Pituitary Cells Following Treatment with Diethyl-Stilbestrol and Medroxy-Progesterone and Their Effects on Ovarian Activity and Hematological Variables of Female Rabbits

    Kaleem Iqbal1, Nafees Akhtar1*, Nazir Ahmad1 and Sajjad-ur-Rahman2


    Full Text Available Aims of this study were: to investigate whether FSH and LH secretion from in-vitro cultured buffalo adenohypophyseal cells can be increased by supplementing culture media with diethyl-stilbestrol and medroxy-progesterone, respectively; to monitor bioactivity of these in-vitro produced gonadotropins and to see if these gonadotropins have any adverse effects on hematology and internal body organs of female rabbits. Pituitary glands collected from 36 adult buffaloes slaughtered at a local abattoir were used. The anterior pituitary cells were cultured in-vitro using medium RPMI-1640 (code R6504-Sigma enriched with 10% fetal calf serum and GnRH and treated with 0.5 or 1.0 mg/100 ml diethyl-stilbestrol, and 2.5 or 5.0 mg/ml medroxy-progesterone, or left as untreated control. The results showed that FSH and LH concentrations from cultures treated with low or high dose of respective steroids were higher (P<0.05 than those for controls. Treatment of pre-pubertal female rabbits with in-vitro extracted FSH increased serum FSH and LH concentrations, ovarian size and number of developing follicles (GFs on the ovaries compared to controls (P<0.01. However, rabbits treated with in-vitro produced extract of LH showed increased serum FSH and LH, while there was no effect on ovarian size and number of GFs. Moreover, treatment of rabbits with both gonadotropins had no effects on body weight, hematological variables and internal body organs. In conclusion, diethyl-stilbestrol and medroxy-progesterone enhanced the secretion of FSH and LH, respectively, from cultured pituitary cells. Moreover, in-vitro produced FSH increased ovarian size, serum FSH and LH and stimulated ovarian activity, while in-vitro produced LH neither increased ovarian size nor stimulated ovarian activity.

  8. Prenatal diethylstilbestrol induces malformation of the external genitalia of male and female mice and persistent second-generation developmental abnormalities of the external genitalia in two mouse strains

    Mahawong, Phitsanu; Sinclair, Adriane; Li, Yi; Schlomer, Bruce; Rodriguez, Esequiel; Max, Ferretti M.; Liu, Baomei; Baskin, Laurence S.; Cunha, Gerald R.


    Potential trans-generational influence of diethylstilbestrol (DES) exposure emerged with reports of effects in grandchildren of DES-treated pregnant women and of reproductive tract tumors in offspring of mice exposed in utero to DES. Accordingly, we examined the trans-generational influence of DES on development of external genitalia (ExG) and compared effects of in utero DES exposure in CD-1 and C57BL/6 mice injected with oil or DES every other day from gestational days 12 to 18. Mice were examined at birth, and on 5 to 120 days postnatal to evaluate ExG malformations. Of 23 adult (≥60 days) prenatally DES-exposed males, features indicative of urethral meatal hypospadias (see text for definitions) ranged from 18 to 100% in prenatally DES-exposed CD-1 males and 31 to 100% in prenatally DES-exposed C57BL/6 males. Thus, the strains differed in the incidence of male urethral hypospadias. Ninety-one percent of DES-exposed CD-1 females and 100% of DES-exposed C57BL/6 females had urethral-vaginal fistula. All DES-exposed CD-1 and C57BL/6 females lacked an os clitoris. None of the prenatally oil-treated CD-1 and C57BL/6 male and female mice had ExG malformations. For the second-generation study, 10 adult CD-1 males and females, from oil- and DES-exposed groups, respectively, were paired with untreated CD-1 mice for 30 days, and their offspring evaluated for ExG malformations. None of the F1 DES-treated females were fertile. Nine of 10 prenatally DES-exposed CD-1 males sired offspring with untreated females, producing 55 male and 42 female pups. Of the F2 DES-lineage adult males, 20% had exposed urethral flaps, a criterion of urethral meatal hypospadias. Five of 42 (11.9%) F2 DES lineage females had urethral-vaginal fistula. In contrast, all F2 oil-lineage males and all oil-lineage females were normal. Thus, prenatal DES exposure induces malformations of ExG in both sexes and strains of mice, and certain malformations are transmitted to the second-generation. PMID

  9. 已烯雌酚与米索前列醇用于绝经后取环的临床效果观察%Observation on clinical effects of Diethylstilbestrol and Misoprostol for IUD removal in postmenopausal women

    韦建巍; 韦汝凤; 吴丽英; 陈林月; 刘冬艳; 陈韩亮; 韦雪燕


    目的:探讨己烯雌酚与米索前列醇用于绝经后取环手术前用药的临床效果.方法:对绝经后要求取环的120例妇女依照就诊的先后顺序分观察组60例和对照组60例,观察组使用己烯雌酚与米索前列醇后行传统常规取环术,对照组直接传统常规取环术,观察两组术中宫颈软化程度、疼痛情况、出血情况、取环时间、取环成功率.结果:己烯雌酚与米索前列醇于绝经后取环手术前用药,能使取环术中宫颈软化满意,疼痛轻,出血量少,取环的时间短,成功率高(P<0.05).结论:己烯雌酚与米索前列醇用于绝经后取环手术前用药安全、有效、方便.%Objective: To explore the clinical effects of Diethylstilbestrol and Misoprostol for intrauterine device (IUD) removal in postmenopausal women as preoperative drugs. Methods; A total of 120 postmenopausal women who demanded IUD removal were divided into observation group and control group according to visiting sequence, 60 women in each group; the postmenopausal women in observation group were treated with routine IUD removal surgery after preoperative medication with Dielhylstilbestrol and Misoprostol', while the postmenopausal women in control group were treated with routine IUD removal surgery; the degrees of cervical softening, pain situations, bleeding situations, removal times, successful rates of IUD removal in the two groups were observed. Results: Diethylstilbestrol and Misoprostol can softened cervix for IUD removal in postmenopausal women as preoperative drugs, the pain was light, the amount of blood loss was few, the removal time of IUD was short, and the successful rate was high (P<0.05) .Conclusion; Diethylstilbestrol and Misoprostol are safe, effective, and convenient for IUD removal in postmenopausal women.

  10. Long-term application of diethylstilbestrol upregulates expressions of μ- and m-calpains in pituitary intermediate lobe of female Wistar rats

    Weijiang Zhao; Fang Yuan; Guilin Li; Zhongfang Shi; Yun Cui; Yazhuo Zhang; Zhongcheng Wang


    BACKGROUND: During formation of prolactin neoplasia, how cells and its structure in adenohypophysis affect prolactin cells should be further studied. Intermediate lobe can be regarded as a driving region to release prolactin (PRL) and may promote formation of prolactin neoplasia in pituitary anterior lobe. OBJECTIVE: To observe the effect of diethylstilbestrol (DES) on the expressions of μ and m-calpains in pituitary intermediate lobe of female Wistar rats. DESIGN: Observational contrast animal study. SETTING: Beijing Neurosurgical Institute.MATERIALS: A total of 21 female Wistar rats, 3 weeks old weighing 70 - 80 g were housed with free access to tap water and standard pellet food. They were kept in a CL-grade condition, at (24±1) ℃ and a humidity of (55±5)%, and with a 12 hours day-night cycle. Caprine anti-μ- and m-calpains antibodies were provided by Santa Cruz Biotechnology, CA, USA; rabbit-anti-PRL antibodies by Dako, Denmark; rabbit-anti-ACTH antibody by Boster Company, Wuhan.METHODS: The experiment was carried out in Pathophysiological Department and Animal Laboratory, Beijing Neurosurgical Institute from August 2006 to January 2007. ①Rats were randomly divided into groups with 7 in each group, including vehicle control group, in which rats were injected intraperitoneally with sun-flower seed oil (1 Ml/kg, twice a week) for 16 weeks; DES group, where animals were administered with DES (5 mg/kg, twice a week) for 16 weeks; DES + vehicle control group, in which DES was administered for 12 weeks at the same dose with those in DES group, and then was discontinued and replaced by sun-flower seed oil (1 Ml/kg, twice a week) for the following 4 weeks. ②At 16 weeks later, pituitary tissue was dealt with HE staining and PRL immunohistochemical examination to observe evoke of tumor; meanwhile, immunohistochemical examination was used to observe expression of PRL of pituitary anterior lobe, expressions ofμ- and m-calpains of pituitary intermediate lobe and

  11. 米曲霉Aspergillus oryzae M-4降解己烯雌酚的特性研究%Degradation Characteristics of Diethylstilbestrol by Aspergillus oryzae M-4

    胡凯弟; 邓维琴; 陈树平; 柴先杜; 刘爱平; 卓文杰; 刘书亮


    以1株分离自酱油曲的米曲霉(Aspergillus oryzae)M-4为材料,初步研究其降解己烯雌酚(diethylstilbestrol,DES)的特性.米曲霉M-4对DES的降解率与菌体生物量呈正相关,在基础盐培养基(mineral salt medium,MM)中培养9d对100mg/L的DES降解率为93%.动力学研究表明,该菌株降解DES的过程符合一级动力学方程,在所测试的培养温度、初始pH值、底物质量浓度范围内,DES半衰期为1.645~5.295d.培养温度30℃和偏酸性环境有利于其对DES的降解;底物质量浓度越高,其半衰期越长.

  12. 分子印迹仿生光子晶体的制备及其在己烯雌酚检测中的应用%The Fabrication of Molecular Imprinted Bionic Photonic Crystal for the Detection of Diethylstilbestrol

    赛娜; 吴蕴棠; 孙忠; 高志贤; 黄国伟


    Molecular imprinted bionic photonic crystals was synthesized and applied in the detection of diethylstilbestrol residues. Molecular imprinted bionic photonic crystals with high performances were fabricated by the suspension polymerization and the vertical settlement. Molecular imprinted bionic photonic crystals showed the high sensitivity (as low as 10 ng/mL)and specificity. Based on the combination of molecular imprinting technique with photonic crystals, a high-performance detection materials, namely molecularly imprinted bionic photonic crystal, can be developed, exhibiting not only simple, inexpensive preparation but also high detection performance.%探讨分子印迹仿生光子晶体的制备及其在己烯雌酚残留检测应用的可行性。方法采用悬浮聚合法、垂直沉降法等方法制备出性能稳定的分子印迹仿生光子晶体。结果分子印迹仿生光子晶体对己烯雌酚残留最低响应浓度为10 ng/mL,并展现出较高的响应特异性。结论将分子印迹技术与光子晶体结合可发展出一种高性能检测材料,即分子印迹仿生光子晶体,不仅具有制备简单、制备材料低廉等优势而且具有较高的检测性能。

  13. The Effect of Diethylstilbestrol on Gubernaculum Testis Development in Fetal Mice%己烯雌酚致胚胎期睾丸引带形态结构发育异常的研究

    邓汪东; 蒋学武; 李建宏; 赖亚曼; 陈中献; 黄天华


    背景与目的:研究环境外源性雌激素己烯雌酚(Diethylstilbestrol,DES)对胚胎小鼠睾丸引带形态发育的影响,探讨DES影响睾丸以及生殖系统发育的机制.材料与方法:昆明雌性小鼠60只,随机分成6组,于孕9~17 d每天分别给予DES 25、50、100、200μg·kg-1·d-1和等体积的DMSO、生理盐水作空白和正常对照.孕19 d处死母鼠,取出活胎、取其下腹部,分别作光镜和电镜固定.观察睾丸引带的组织形态学及细胞超微结构变化.结果:DES可致睾丸引带发育不良:体积缩小,形态异常,组织结构紊乱,细胞内肌丝细小,排列紊乱,密体不明显,胞浆中细胞器散在分布.DES剂量越大,睾丸引带形态异常及组织细胞结构紊乱现象越明显.结论:DES可致胎鼠睾丸引带形态异常、组织细胞结构紊乱,且有剂量效应关系.

  14. Effect of in utero exposure to diethylstilbestrol on lumbar and femoral bone, articular cartilage, and the intervertebral disc in male and female adult mice progeny with and without swimming exercise.

    Rowas, Sora Al; Haddad, Rami; Gawri, Rahul; Al Ma'awi, Abdul Aziz; Chalifour, Lorraine E; Antoniou, John; Mwale, Fackson


    Developmental exposure to estrogens has been shown to affect the musculoskeletal system. Furthermore, recent studies have shown that environmental exposure to estrogen-like compounds is much higher than originally anticipated. The aim of this study was to determine the effects of diethylstilbestrol (DES), a well-known estrogen agonist, on articular cartilage, intervertebral disc (IVD), and bone phenotype. C57Bl/6 pregnant mice were dosed orally with vehicle (peanut oil) or 0.1, 1.0, and 10 μg/kg/day of DES on gestational days 11 to 14. Male and female pups were allowed to mature without further treatment until 3 months of age, when swim and sedentary groups were formed. After euthanasia, bone mineral density (BMD), bone mineral content (BMC), bone area (BA), and trabecular bone area (TBA) of the lumbar vertebrae and femur were measured by using a PIXImus Bone Densitometer System. Intervertebral disc proteoglycan was measured with the DMMB assay. Histologic analysis of proteoglycan for IVD and articular cartilage was performed with safranin O staining, and degeneration parameters were scored. The lumbar BMC was significantly increased in female swimmers at both the highest and lowest dose of DES, whereas the femoral BMC was increased only at the highest. The males, conversely, showed a decreased BMC at the highest dose of DES for both lumbar and femoral bone. The female swim group had an increased BA at the highest dose of DES, whereas the male counterpart showed a decreased BA for femoral bone. The TBA showed a similar pattern. Proteoglycan analysis of lumbar IVDs showed a decrease at the lowest doses but a significant increase at the highest doses for both males and females. Histologic examination showed morphologic changes of the IVD and articular cartilage for all doses of DES. DES significantly affected the musculoskeletal system of adult mice. Results suggest that environmental estrogen contaminants can have a detrimental effect on the developmental lumbar


    谭号; 李英文; 尹盼; 刘智皓


    Diethylstilbestrol (DES) is a typical endocrine disruptor for aquatic animals in the Yangtze River of China. Here we investigated the effects of DES on testicular development and spermatogenesis of fish. Adult male zebrafish (Danio rerio) were used as experimental subjects and were exposed to DES (0.1, 1 and 10μg/L) for 20 days. The histo-logical results demonstrated that the DES exposure led to severe impacts on zebrafish spermatogenesis. To further elu-cidate mechanisms underlying this phenomenon, we cloned the cDNAs of vasa and dmc1 and analyzed their expression patterns at the tissue and cellular levels. Our results showed that vasa was exclusively expressed in spermatogonia and primary spermatocytes of testis, and that dmc1 was specifically expressed in spermatocytes of testis. Using semi-quantitative RT-PCR we found that DES dramatically suppressed the expressions of dmc1 in a dose- and time-dependent manner, but did not affect the expression of vasa. Moreover, the expression of dmrt1 (the male sex de-termining gene) and P450 11β(the key enzyme responsible for 11-KT synthesis) were also suppressed by DES exposure. Given that these genes play a role in meiosis and spermatogenesis, we speculated that DES might induce the male germ cell apoptosis in zebrafish by suppressing the expression of dmrt1 and P450 11β, and might inhibit the expression of dmc1 which result in impeded meiosis.%为研究内分泌干扰物己烯雌酚(DES)对鱼类精巢发育和配子发生的影响,研究用DES(0.1、1和10µg/L,暴露20d)对内分泌干扰研究的经典模式动物——斑马鱼(Danio rerio)雄性成鱼进行了处理。组织学研究结果表明, DES严重影响斑马鱼精子发生。同时,研究克隆了斑马鱼与生殖细胞发育和减数分裂相关的vasa、dmc1的部分cDNA,对其组织和细胞表达模式进行了研究。结果表明, vasa仅表达于精巢的精原细胞、初级精母细胞和卵巢不同时期的生殖细胞;而dmc1则表达于

  16. The expression of estrogen receptor genes in diethylstilbestrol-induced pituitary prolactinoma in rats%雌激素受体基因在乙菧酚诱发的大鼠催乳素瘤中的表达

    徐春; 李江源; 黄兆坚


    目的 研究雌激素受体(ER)α、ERβ和断裂的雌激素受体产物-1(TERP-1)在乙酚(DES)诱发的大鼠催乳素瘤中的表达。方法 雌性Wistar大鼠摘除卵巢后皮下植入含DES 20 mg的硅胶管,以空白硅胶管作对照。8周后处死动物,用放免法测定大鼠血清催乳素水平,观察大鼠垂体组织学改变,并用免疫组化方法检测催乳素染色阳性细胞,用逆转录-聚合酶链反应分析垂体组织中ERα、ERβ和TERP-1 mRNA的转录水平。结果 实验组大鼠血清催乳素水平、垂体重量和垂体组织催乳素阳性细胞计数均明显高于对照组(P<0.001),ERα、ERβ和TERP-1 mRNA在两组大鼠垂体组织中均有转录,其中ERα和TERP-1 mRNA在实验组中的转录水平比在对照组高(P<0.001和P<0.05)。结论 雌激素可以通过雌激素受体直接影响垂体催乳素细胞。正常的雌激素受体与异常的雌激素受体的共同存在提示雌激素调节作用的异质性,它们之间的相互关系及与垂体肿瘤发生的关系,有待于进一步阐明。%Objective To examine the expressions of estrogen receptor α (ERα), estrogen receptor β (ERβ) and pituitary-specific truncated estrogen receptor product-1 (TERP-1) in diethylstilbestrol (DES) induced prolactinoma of rats. Methods Ovariectomised Wistar rats were subcutaneously implanted with an implant containing 20 mg DES. Rats were implanted with a blank implant as controls. Eight weeks later, serum prolactin level of rats was measured by RIA. Prolactin immunoreactive cells of anterior pituitary were detected by immunohistochemistry assay. ERα, ERβ and TERP-1 mRNAs in pituitaries were examined by RT-PCR. Results Serum prolactin levels, pituitary weights and pituitary prolactin immunoreactive cell countings in experimental group rats were all obviously higher than those in control group (P<0.001 respectively). ERα, ERβ and TERP-1 mRNAs were all transcripted in DES

  17. Effects of diethylstilbestrol on testicular oxidative stress and steroidogenesis in male rats%己烯雌酚诱导的氧化应激对青春期大鼠睾丸类固醇合成的影响

    李军延; 乔佩环; 张林媛; 刘帅; 于淼; 常兵


    known that diethylstilbestrol ( DES ) can result in testicular oxidative injury , and one of its mechanisms of action is leading to dysfunction of steroidogenesis .The aim of this study was to investigate the relationship between testicular oxidative injury caused by DES and the key synthetase activities for the synthesis pathway of steroidogenesis and the possible mechanism .Methods Twenty-four 4-wk-old male Wistar albino rats were randomly divided into 4 groups , 6 rats each.Three doses of DES (0.1, 1.0 and 10 μg/kg· d) groups and a vehicle (corn oil) control group , were respectively administered by subcutaneous injection once a day for eight weeks .The rats were sacrificed after 8 weeks treatment and the body weight , testis, epididymis, prostate were weighed, respectively.The testicular tissues were homogenized and the oxidation of MDA and ROS , the activity changes of antioxidases SOD, CAT and GPx, as well asthe activities of steroid synthetases 3β-HSD1 and 17β-HSD3 were determined by biochemical measurement.The levels oftestosterone and LH in peripheral blood were measured by radioimmunoassay .The intensities of expression of StAR,P450scc, 3β-HSD1, 17β-HSD3-mRNA were detected by PCR.Results In the 10.0 μg/kg dose group, the weights andorgan coefficients of testis and prostate were decreased significantly , the oxidation of MDA and ROS was increased distinctlyand the activities of SOD, CAT, GPx, 3β-HSD1 and 17β-HSD3 were reduced.The concentration of serum testosterone wasdecreased in the 10.0 μg/kg dose group.In the 10.0 μg/kg and 1.0 μg/kg dose groups, the decline of LH levelpresented a dose-dependent manner, and the intensities of immunochemical positive staining for StAR , P450scc, 3β-HSD1and 17β-HSD3 mRNA were decreased.Conclusions DES exposure results in disturbance of the oxidant /antioxidantbalance and decline of testosterone level that induces reproductive impairment in male rats .DES induces reductions of bothGPx and 3β-HSD activities which

  18. 己烯雌酚与淫羊藿总黄酮联合用药对去卵巢大鼠腰椎和股骨的影响%Influence of diethylstilbestrol combined with epimedium pubescens flavonoids on lumbar vertebral body and femur in ovariectomized rats

    许碧连; 吴铁; 崔燎; 刘钰瑜; 邹丽宜


    BACKGROUND: Previous experiments indicate that epimedium pubesceus flavonoids (EPF) could enhance protective effect of estrogen on proximal tibial metaphysis (PTM) and reduce its adverse effect on uterine tissue in ovariectomized rats. Do they have synergetic effects on lumbar vertebral body and femur in ovariectomized rats when used in combination?OBJECTIVE: To observe the influence of diethylstilbestrol combined with EPF on metabolic biochemical indexes and bone histomorphometry of lumbar and femur in ovariectomized rats.DESIGN: Randomized controlled study on experimental animals.SETTING: Department of Pharmacology, Guangdong Medical College.MATERIALS: Female unmated SD rats of 4 months old, with the body mass of (225±15)g, were provided by Shanghai Experimental Animal Centre of the Chinese Academy of Sciences. They were of clean grade (Certification No. CSAM003). All rats were randomized into 5 groups: namely,sham- operation group, ovariectomy group, diethylstilbestrol group, EPF group, and combination group with 10 rats in each group.METHODS: Rats in all the groups except sham operation group were subjected to bilateral ovariectomy. Those in sham operation group and ovariectomy group were given solvent as control, while rats in the other groups were given gastric perfusion of: diethylstilbestrol at the dosage of 22.5 μg/metric analysis was used to assess the static parameters [percent trabecular area(%Tb. Ar), trabecular number(Tb.N), trabecular thickness (Tb.Th), trabecular seperation(Tb. Sp), osteoclast number per mm2 (Oc.N)] and dynamic parameters[percent labeled perimeter(%L.Pm), mineral apposition rate (MAR), bone formation rate(BFR)per unit of bone volume (BFR/BV)] of the fifth lumbar vertebral body(LV5). Meanwhile the contents of femur calcium,phosphorus and hydroxyproline were also measured.static parameters of LV5 of ovariectomized rats due to the combined adethylstilbestrol and EPF on femur biochemical indexes in ovariectomized rats.RFSULTS: Five

  19. Determination of trace diethylstilbestrol in urine by hollow fiber membrane/liquid-phase microextraction combined with HPLC%中空纤维膜液相微萃取-高效液相色谱联用技术测定尿液中痕量己烯雌酚

    张玉; 刘雷英


    采用中空纤维液相微萃取与高效液相色谱联用技术测定了尿液样品中的痕量己烯雌酚;考察了样品相酸度、中间相种类、接收相浓度、搅拌速度、萃取时间等对液-液-液三相微萃取效率的影响,进而确定了最佳萃取条件.结果表明,当样品相pH为2.5,中间相为甲苯,接收相为3μL 0.25 mol/L氢氧化钠溶液,搅拌速度为800r/min,萃取时间为50 min时,萃取效率最佳.在最佳萃取条件下,样品的回收率为76.4%,相对标准偏差为3.8%.%Trace diethylstilbestrol in urine samples was determined by combining hollow fiber membrane/liquid-phase microextraction with high performance liquid chromatography. The effects of donor phase pH, intermediate type, acceptor phase concentration, stirring rate, and extraction time on liquid-liquid-liquid tri-phase microextraction efficiency were investigated, and the optimal microextraction condition was established accordingly. Results indicate that, when toluene is used as the intermediate phase and 3 μL of 0. 25 mol/L NaOH solution as the acceptor phase, the best microextraction efficiency is obtained after 50 min of extraction under a stirring rate of 800 r/min. The recovery of the tested urine samples is 76. 4% and the relative standard deviation is 3. 8% under the optimized microextraction condition.

  20. Voltammetric behavior of diethylstilbestrol at conductive carbon black paste electrode and its application in trace amount detection%己烯雌酚在导电炭黑糊电极上的伏安行为及其在痕量检测中的应用

    李猛; 张旭志; 张艳; 马绍赛; 崔毅; 曲克明


    Conductive carbon black paste electrodes ( CCBPEs) displayed excellent electroanalytical ability due to the high signal/ noise ratio. The electrochemical behavior of diethylstilbestrol( DES) was studied by cyclic voltammetry( CV)and linear sweep volta-mmetry( LSV). The mechanism of electrode reaction was investigated preliminarily. Based on the results,some of important factors, including species of the supporting electrolyte, pH, accumulation potential and accumulation time, which had effect on the voltammetric response signal,were studied and optimized. Under the optimized conditions,there was a linear relationship between the oxi-dative peak value and the concentration of DES in the range of 3. 1 x 10~9 ?. 55 x 10~7 mol/L with a detection limit of 1. 55xlO~' mo)/L(signal/noise = 3). Then a rapid and highly sensitive detection method for DES was established. Satisfactory results were obtained when the new method was used to measure DES in the real fishery water samples.%由于具有较高的信/噪比,导电炭黑糊电极(CCBPE)具有优良的电分析化学性能.论文采用循环伏安(CV)及线性扫描伏安(LSV)等方法研究了己烯雌酚(DES)在CCBPE上的电化学行为,初步探讨了电极响应机理.在此基础上,对影响伏安响应信号的因素(诸如支持电解质种类、pH值、富集电位和富集时间等)进行了研究和优化.在优化条件下,DES的氧化峰电流与其浓度在3.1×10-9 ~ 1.55×10-7 mol/L范围内呈良好的线性关系,检测限为1.55×10-9mol/L( S/N=3).据此建立了一种快速、高灵敏的DES检测方法.应用于测定经过简单预处理的淡水渔业水样,得到了满意的结果.

  1. [Diethylstilbestrol exposure in utero. Polemics about metroplasty. The cons].

    Epelboin, S


    Uterine malformations in DES-exposed women are not the only aetiologies for infertility, miscarriages, and other problems in their reproductive life. A global screening of fertility factors of the couple may, for instance, show in them vascular uterine abnormalities which reduce their reproductive potential. Furthermore, these abnormalities are not always predictive of losses of pregnancy, and many exposed women with patent uterine abnormalities can carry a pregnancy to term. Metroplasty for uterine enlargement is a surgical procedure suggested for restoring the size and shape of the uterine cavity. There are no comparative studies for assessing efficacy and safety of metroplasty. Therefore, metroplasty should not be performed routinely, but should only be considered after the couple has undergone a full fertility workup, and the best possible level of fertility has been achieved.

  2. 氧化石墨烯/壳聚糖修饰玻碳电极同时测定己烯雌酚和辛基酚%Determination of diethylstilbestrol and octylphenol on oxide graphene/chitosan modified glassy carbon electrode at the same time

    季万余; 于春梅; 李晓东; 何红; 顾海鹰


    Objective:To establish an electrochemical determination of diethylstilbestrol(DES) and octylphenol(OP) at the same time . Methods: Through self-assembly methods , graphene oxide ( GO ) was assembled to chitosan modified glassy carbon electrode surface by selfassembly to build the GO-chitosan modified glassy carbon electrodes, in phosphate buffer solution of pH 7.0, we used the method of cyclic voltammetry and differential pulse voltammetry to study the method and admeasurement of electrochemical action and determine DES and OP on the modified electrodes. Origin8 software multi-peak curve fitting analysis was applied to analyse the data of experiment. Results: In the solution of pH 7.0, the linearity range of DES and OP were 3.31 ×10-7-1.24 ×10-5 mol/L and 2.89 ×10-7-9.50 ×10-6 mol/L. Detection limit is 7.56 ×10-8 mol/L, it was indicated that modified electrode had good repeatability and stability. Conclusion: Determination of phenolic environment estrogen in GO-chitosan modified glassy carbon electrodes is simple but high-speed, can test the composite sample of DES and OP.%目的::建立同时测定己烯雌酚(diethylstilbestrol,DES)和辛基酚(octylphenol,OP)的电化学方法。方法:通过自组装技术将氧化石墨烯(graphene oxide,GO)组装到壳聚糖修饰的玻碳电极表面,构建GO-壳聚糖修饰玻碳电极,用循环伏安法和差分脉冲伏安法研究DES和OP在该修饰电极上的电化学行为及其测定,利用Origin8软件多峰曲线拟合分析实验数据。结果:在pH 7.0磷酸盐缓冲溶液中,同时测定DES和OP的线性范围分别为3.31×10-7~1.24×10-5 mol/L和2.89×10-7~9.50×10-6 mol/L,检测限为7.56×10-8 mol/L。且该修饰电极具有良好的重复性和稳定性。结论:GO/壳聚糖修饰玻碳电极测定酚类环境刺激素具有简单、快速等优点,可同时测定DES和OP的混合样品。

  3. 黄酮和维生素E对己烯雌酚所致雄性小鼠肝脏氧化损伤的保护作用%Protection Effects of Flavonoids and Vitamin E on Diethylstilbestrol Induced Oxidative Injury

    杜娟; 潘红艳; 林利美; 黄大伟; 宫智勇


    本试验旨在探讨黄酮(flavonoids)和维生素E(VE)对己烯雌酚(diethylstilboestrol,DES)所致昆明雄性小鼠肝脏氧化损伤的保护作用.将80只小鼠随机分为正常对照组、DES模型组(0.035、0.35、3.5 mg/(kg·d))、黄酮干预组(26、58、112 mg/(kg·d))和VE干预组(56.2、112.4、224.8 mg/(kg· d));干预组先灌服3.5 mg/(kg,d)DES处理,再灌服不同剂量黄酮和VE;正常对照组灌服等体积生理盐水.灌胃给药,连续7d,试验结束后禁食24 h,脱颈处死,剖检肝脏和睾丸.计算肝脏和睾丸脏器系数,测定肝脏组织中的丙二醛(MDA)含量、总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性.结果表明,不同剂量的DES组小鼠肝脏系数和睾九系数均下降;112.0 mg/(kg·d)黄酮剂量组与高剂量DES组相比,SOD活性上升,MDA极显著下降(P<0.01);与高剂量DES组相比,112.4、224.8 mg/(kg·d)VE剂量组T-AOC极显著上升(P<0.01),MDA极显著下降(P<0.01).224.8 mg/(kg·d)VE剂量组GSH-PX极显著上升(P<0.01).不同浓度的DES均导致肝脏不同程度氧化损伤,黄酮和VE均能提高机体的抗氧化能力,降低DES对机体造成的氧化损伤.%The purpose of this study was to investigate the oxidative damage caused by different doses of diethylstilbestrol (DES) and the protective effects for DES induced toxicity with flavonoids and vitamin E in Kunming mice. 80 Kunming male mice were randomly divided into 10 groups, the groups were normal saline control group, DES exposed groups (0. 035, 0. 35, 3.5 mg/(kg·d)), flavonoids interference groups(26. 0. 58.0, 112. 0 mg/(kg·d)) and vitamin E interference groups( 56. 2, 112. 4, 224. 8 mg/(kg·d)). The effects on liver and testis were studied in adult male mice treated for one week. Mice were killed to collect liver and testis, and then contents of malonadehyde (MDA), superoxide dismutase (SOD), glutathione perxi-dase (GSH-Px) and total antioxidant capacity (T

  4. Gynaecomastia linked to the intake of a herbal supplement fortified with diethylstilbestrol

    Toorians, A.W.F.T.; Bovee, T.F.H.; Rooy, De J.; Stolker, L.A.A.M.; Hoogenboom, L.A.P.


    This study reports the findings of a supplement marketed on the Internet for prostate problems. The supplement was orally taken by a 60-year-old man with divergent hormonal levels and who was surgically treated for gynaecomastia: development of abnormally large mammary glands in males. The supplemen


    Five natural, pharmaceutical, or xenobiotic chemicals (17b-estradiol, ethynylestradiol, diethystilbestrol, nonylphenol, methoxychlor) were tested in two in vitro (MCF-7 breast tumor cell proliferation [E-screen], yeast estrogen system [YES]), and one in vivo (male sheepshead min...

  6. Degradation of Diethylstilbestrol in Aqueous Solution by Ozonation%水溶液中己烯雌酚的臭氧氧化研究

    张霞; 冯精兰; 吴峰; 邓南圣


    利用臭氧氧化技术对己烯雌酚(DES)水溶液进行了研究,考察DES溶液初始浓度、溶液初始pH值、臭氧投加量对DES降解的影响.结果表明,在pH3.0~9.0范围内,pH初始值越高,DES降解率越大;在5~20 mg/L范围内,DES初始浓度越大,降解效率越高,反应属于一级动力学,动力学方程为-dc/dt=0.087 85 c;臭氧投加量越大,DES去除率越高;HPLC色谱图表明,在DES臭氧氧化降解过程中,有极性较DES大的产物;氧化过程中溶液的pH值和电导率的变化结果表明,有小分子酸性物质产生.

  7. The endocrine disruptors dibutyl phthalate (DBP) and diethylstilbestrol (DES) influence Leydig cell regeneration following ethane dimethane sulphonate treatment of adult male rats

    Heng, K.; Anand-Ivell, R.; Teerds, K.J.; Ivell, R.


    The manner by which endocrine-disrupting xenobiotics, such as phthalates, can induce changes in the development of the male reproductive system still remains largely unknown. Herein, we have explored the application of ethane dimethane sulphonate (EDS) to eliminate adult-type Leydig cells in the mat

  8. Early exposure of 17α-ethynylestradiol and diethylstilbestrol induces morphological changes and alters ovarian steroidogenic pathway enzyme gene expression in catfish, Clarias gariepinus.

    Sridevi, P; Chaitanya, R K; Prathibha, Y; Balakrishna, S L; Dutta-Gupta, A; Senthilkumaran, B


    Environmental estrogens are major cause of endocrine disruption in vertebrates, including aquatic organisms. Teleosts are valuable and popular models for studying the effects of endocrine disrupting chemicals (EDCs) in the environment. In the present study, we investigated the changes caused by exposure to the synthetic estrogens 17α-ethynylestradiol (EE2 ) and diethylstilbesterol (DES) during early stages of growth and sex differentiation of air-breathing catfish, Clarias gariepinus, at the morphological, histological, and molecular levels. Catfish hatchlings, 0 day post hatch (dph) were exposed continuously to sublethal doses of EE2 (50 ng/L) and DES (10 ng/L) until 50 dph and subsequently monitored for ovarian structural changes and alteration in the gene expression of steroidogenic enzymes till adulthood. Treated fish exhibited morphological deformities such as spinal curvature, stunted growth, and yolk-sac fluid retention. In addition to ovarian atrophy, DES-treated fish showed either rudimentary or malformed ovaries. Detailed histological studies revealed precocious oocyte development as well as follicular atresia. Further, transcript levels of various steroidogenic enzyme and transcription factor genes were altered in response to EE2 and DES. Activity of the rate-limiting enzyme of estrogen biosynthesis, aromatase, in the ovary as well as the brain of treated fish was in accordance with transcript level changes. These developmental and molecular effects imparted by EE2 and DES during early life stages of catfish could demonstrate the deleterious effects of estrogen exposure and provide reliable markers for estrogenic EDCs exposure in the environment.

  9. Organizational effects of perinatal exposure to bisphenol-A and diethylstilbestrol on arcuate nucleus circuitry controlling food intake and energy expenditure in male and female CD-1 mice.

    Mackay, Harry; Patterson, Zachary R; Khazall, Rim; Patel, Shoyeb; Tsirlin, Dina; Abizaid, Alfonso


    The endocrine disrupting compound bisphenol-A (BPA) has been reported to act as an obesogen in rodents exposed perinatally. In this study, we investigated the effects of early-life BPA exposure on adult metabolic phenotype and hypothalamic energy balance circuitry. Pregnant and lactating CD-1 dams were exposed, via specially prepared diets, to 2 environmentally relevant doses of BPA. Dams consumed an average of 0.19 and 3.49 μg/kg per day of BPA in the low and high BPA treatments prenatally and an average of 0.36 and 7.2 μg/kg per day of BPA postnatally. Offspring were weaned initially onto a normal (AIN93G) diet, then as adults exposed to either a normal or high-fat diet (HFD). Males exposed to the high dose of BPA showed impaired glucose tolerance on both diets. They also showed reduced proopiomelanocortin fiber innervation into the paraventricular nucleus of the hypothalamus, and when exposed to HFD, they demonstrated increased neuropeptide Y and Agouti-related peptide expression in the arcuate nucleus (ARC). Females exposed to the high BPA dose were heavier, ate more, and had increased adiposity and leptin concentrations with reduced proopiomelanocortin mRNA expression in the ARC when consuming a HFD. BPA-exposed females showed ARC estrogen receptor α expression patterns similar to those seen in males, suggesting a masculinizing effect of BPA. These results demonstrate that early-life exposure to the obesogen BPA leads to sexually dimorphic alterations in the structure of hypothalamic energy balance circuitry, leading to increased vulnerability for developing diet-induced obesity and metabolic impairments, such as glucose intolerance.

  10. Clear cell adenocarcinoma of the cervix in second generation young women who are without maternal exposure to diethylstilbestrol: A case report

    Tanitra Tantitamit


    Full Text Available Clear cell adenocarcinoma of the cervix (CCAC is a rare type of gynecological cancer. The risk factors and pathogenesis have yet to be clearly determined. This is a case report of a 19-year-old woman, who was never exposed to DES from her mother, who had gone for an examination for vaginal bleeding. A polypoid mass measured at 3 cm presenting in her cervix was discovered. Histological diagnosis following cervical biopsy proved the tumor to be a clear cell carcinoma. The patient was then referred to our hospital. A loop electrosurgical excision procedure (LEEP was done and the pathological diagnosis was confirmed for clear cell carcinoma with a positive endocervical margin. Radical hysterectomy, pelvic lymphadenectomy and incidental appendectomy were achieved without any complications. The microscopic findings had revealed no residual tumor. The patient did not require adjuvant treatment. No sign of recurrence has been detected throughout 6 months of follow-up. We have performed the literature review on the clinical presentation, pathogenesis, pathology, diagnosis, treatment, and prognosis of this unusual tumor.

  11. The Preliminary Study on the Embryotoxic Effect of Diethylstilbestrol in Rats%己烯雌酚对大鼠胚胎毒性的研究

    吴婧; 吴焱森; 赵长瑶


    目的 探讨己烯雌酚对大鼠的胚胎毒性作用.方法 将交配成功的雌鼠在整个妊娠期每天注射己烯雌酚.观察己烯雌酚对妊娠雌鼠及胎鼠的影响.结果 染毒剂量为50μg·kg-1·d-1时,胎鼠体重明显下降.随着染毒剂量的增加雄仔鼠体重逐渐降低,染毒剂量达100μg·kg-1·d-1时,平均窝仔数明显减少,胎鼠身长和尾长也发生显著性的改变.结论 己烯雌酚具有一定的胚胎毒性作用.

  12. Drug: D00946 [KEGG MEDICUS

    Full Text Available D00946 Drug Fosfestrol (JAN/INN); Diethylstilbestrol diphosphate (USP);, active substance: Diethylstilbestrol [DR:D00577] estrogen receptor 1 agonist [HSA:2099] [KO:K08550]; est... OF THE GENITAL SYSTEM G03C ESTROGENS G03CB Synthetic estrogens, plain G03CB02 Diethylstilbest...rol D00946 Fosfestrol (JAN/INN); Diethylstilbestrol diphosphate (USP) G03CC Estrogens, combinat...ions with other drugs G03CC05 Diethylstilbestrol D00946 Fosfestrol (JAN/INN); Diethylstilbestrol diphosphate

  13. Drug: D00577 [KEGG MEDICUS

    Full Text Available D00577 Drug Diethylstilbestrol (USP/INN); Stilbestrol (TN) C18H20O2 268.1463 268.35... MODULATORS OF THE GENITAL SYSTEM G03C ESTROGENS G03CB Synthetic estrogens, plain G03CB02 Diethylstilbestrol D00577 Diethylstilbest...rol (USP/INN) G03CC Estrogens, combinations with other drugs G03CC05 Diethylstilbes...trol D00577 Diethylstilbestrol (USP/INN) L ANTINEOPLASTIC AND IMMUNOMODULATING AGEN...TS L02 ENDOCRINE THERAPY L02A HORMONES AND RELATED AGENTS L02AA Estrogens L02AA01 Diethylstilbestrol D00577 Diethylstilbest

  14. 己烯雌酚对去卵巢大鼠骨钙、骨羟脯氨酸含量的影响%Effects of diethylstilbestrol on bone calcium and bone hydroxyproline c ontents in ovariectomized rats

    刘晓青; 崔燎; 吴铁


    @@己烯雌酚对去卵巢大鼠骨丢失的防治作用,其骨形态计量学参数变化已有许多报道 [1] ,但己烯雌酚对骨钙和骨羟脯氨酸含量的影响至今未见报道。骨钙与骨羟脯氨酸含量的比例 完 整体现了骨质量[2],骨钙增多而骨有机质减少,骨的脆性增加,骨质量不高。骨 羟脯氨酸测定可预测骨有机质的含量[3] 。本研究通过对大鼠骨钙和骨羟脯氨酸含 量测定,观察不同剂量的己烯雌酚对去卵巢大鼠骨钙、骨羟脯氨酸的影响,以进一步研究己 烯雌酚抗骨质疏松的作用机制。

  15. 己烯雌酚单克隆抗体细胞株的筛选及间接ELISA法的建立%Study on preparation of monoclonal antibodies against diethylstilbestrol and development of indirect competitive inhibition ELISA

    单国强; 刘雅红; 康彦龙


    以自行合成的己烯雌酚(DES)免疫原免疫BALB/c小鼠,采用细胞融合、显微克隆等单克隆抗体技术,制备了5株抗DES单克隆抗体分泌杂交瘤细胞株(7B6、4B10、1G10、4E4、2C7);选择其中的7B6株抗体,建立了间接竞争ELISA法(indirect competitive inhibition ELISA),通过初步优化,其检测限为10pg/孔,与同类二苯乙烯类雌激素的交叉反应高,而与天然雌激素雌二醇几乎无交叉反应性.这项研究为DES残留检测试剂盒的开发打下了基础.

  16. Ocorrência de resíduos de dietilestilbestrol e zeranol em fígado de bovinos abatidos no Brasil Occurence of diethylstilbestrol and zeranol residues in liver of bovines slaughtered in Brazil

    Olga Maria Chaves CARDOSO


    Full Text Available O uso de substâncias anabolizantes, de natureza hormonal ou não, é muito difundida na pecuária de corte dos países maiores produtores de carne bovina (EUA, Austrália, Argentina, Canadá, etc.. Dentre estas destacam-se o banido dietilestilbestrol (DES e o controlado zeranol, que aumentam o ganho de peso vivo, o peso da carcaça, a eficiência alimentar e o percentual de carne. O uso porém, pode ocasionar a presença de resíduos nos tecidos e órgãos dos animais que são utilizados como alimento. A presença de resíduos representa um perigo potencial para a saúde humana, o que levou vários países, inclusive o Brasil, a proibirem a utilização destes produtos. O objetivo do presente trabalho foi o de verificar, se a carne colhida no período de julho de 1993 a novembro de 1994, em matadouros frigoríficos pertencentes a Lista Geral dos Exportadores, atende a legislação vigente quanto ao uso destes anabolizantes. Para isto, foram analisadas por radioimunoensaio, 416 amostras de fígado para pesquisa de DES e 385 para zeranol. Observou-se que o DES não foi detectado em nenhuma das amostras (p > 0,05, enquanto que o zeranol foi detectado em duas (p The use of hormonal or non-hormonal agents in livestock is widespread among the greatest meat producers, as the U.S.A., Australia, Canada, Argentina and others. These agents include diethylstibestrol (DES, which is banned, and zeranol, which is controlled. Both substances improve live weight gain, carcass weight, feed conversion and meat percentage. Their use, however, may cause residues presence in tissues and organs of livestock, which, in turn, represents a potential hazard for human health. For this reason, many countries, including Brazil, do not allow their use. This trial aimed at verifying whether beef collected between July 1993 and November 1994 in Brazilian slaughterhouses was in accordance with the current legislation in regard to the above mentioned anabolic substances. Four hundred and sixteen samples of liver were searched for DES and 385 for zeranol presence by radioimmunoassay (RIA. DES was detected in no samples (p > 0,05 on the other hand zeranol was detected in two (p < 0,05. Zeranol presence was confirmed by gas cromatography-mass spectrometry (p < 0,05. Average recoveries obtained for DES were 62,6% ± 5,7 in the extractive phase (³H-DES and 83,8% ± 16,8 in the extractive phase plus RIA (DES. As to zeranol, average recoveries were 63,0% ± 5,8 in the extractive phase (³H-zeranol and 94,8% ± 13,8 in the extractive phase plus RIA (zeranol. Therefore, it is concluded that Brazilian cattle meat is in accordance with current legislation in regard to DES; it is not, however, in regard to zeranol for it was found in 0,52% of samples.

  17. Drug: D07826 [KEGG MEDICUS

    Full Text Available D07826 Drug Hexestrol diphosphate sodium; Diethylstilbestrol diphosphate tetrasodiu...X HORMONES AND MODULATORS OF THE GENITAL SYSTEM G03C ESTROGENS G03CB Synthetic estrogens, plain G03CB02 Diethylstilbest...with other drugs G03CC05 Diethylstilbestrol D07826 Hexestrol diphosphate sodium L ANTINEOPLASTIC AND IMMUNOM...10] Nuclear receptors Estrogen like receptors Estrogen receptor estrogen receptor 1 [HSA:2099] [KO:K08550] Diethylstilbest...rol [ATC:G03CB02 G03CC05 L02AA01] D07826 Diethylstilbestrol diphosphate tetrasodium salt estro

  18. What Are the Risk Factors for Childhood Leukemia?

    ... for a possible link to childhood leukemia include: Exposure to electromagnetic fields (such as living near power lines) Living near a nuclear power plant Infections early in life Mother’s age when child is ... history Fetal exposure to hormones such as diethylstilbestrol (DES) or birth ...

  19. Clearcellekarcinom i livmoderhalsen hos en 17-årig pige

    Rønneberg, Elisabeth Thal; Lidang, Marianne; Palle, Connie


    Clear cell adenocarcinoma of the uterine cervix (CCEA) is a rare disease, accounting for only 1% of all cervical cancers. The disease in young women is linked to diethylstilbestrol (DES) exposure in utero. Following the ban of DES in 1979, CCEA rarely occurs in young women, but still remains a ch...

  20. Clomiphene and hypospadias on a detailed level : Signal or chance?

    Meijer, W.M.; de Jong-van den Berg, L.T.W.; van den Berg, M.D.; Verheij, J.B.G.M.; de Walle, H.E.K.


    BACKGROUND: Clomiphene, a drug used to induce ovulation, is chemically related to diethylstilbestrol. (IDES). DES is associated with vaginal cancer and infertility among daughters and with hypospadias among second-generation male offspring. Because clomiphene has a long half-life and metabolites hav

  1. Altered resistance to Trichinella spiralis infection following subchronic exposure of adult mice to chemicals of environmental concern

    Luebke, R.W.


    The effects of subchronic chemical exposure on expulsion of adult Trichinella spiralis from the small intestine of mice and encystment of newborn larvae in the host's musculature were investigated. Exposure to diethylstilbestrol, benzo(a)pyrene, tris-(1,3-dichloro-2-propyl) phosphate, cyclophosphamide, phorbol myristate acetate, and dimethylvinylchloride prior to infection of mice with 200 infective larvae resulted in larger worm burdens in treated animals than in controls 14 days after infection. Worm expulsion was not affected by exposure to tris-(2,3-dibromopropyl)phosphate, orthophenylphenol, and indomethacin. Increased burdens of muscle-phase larvae were found in animals that maintained significant numbers of adult worms in the gut at 14 days, except in mice administered diethylstilbestrol and dimethylvinylchloride. Exposure to diethylstilbestrol and cyclophosphamide resulted in decreased inflammatory reactions in the tissues of the small intestine and development of bone marrow eosinophilia in infected mice. Marrow eosinophilia was likewise decreased in mice given tris-(1,3-dichloro-2-propyl)phosphate before infection. Additional studies with diethylstilbestrol administered either before, at the time of, or after infection showed inhibition of worm expulsion. Drug exposure during a primary infection inhibited the expulsion of a second T. spiralis infection, but did not affect worm elimination when given during a second infection. Treatment with diethylstilbestrol after artificial sensitization of mice with Trichinella antigens decreased delayed hypersensitivity responses to the sensitizing antigen. Immune functions, assessed by lymphoproliferative responses to mitogens and antibody responses to sheep red blood cells, generally correlated with altered host resistance to T. spiralis infection.

  2. Metabolic effects of human growth hormone and of estrogens in boys with Duchenne muscular dystrophy.

    Rudman, D; Chyatte, S B; Patterson, J H; Gerron, G G; O'Beirne, I; Barlow, J; Jordan, A; Shavin, J S


    Metabolic balance studies were conducted in seven boys with Duchenne-type muscular dystrophy, and in six normal boys of similar age, during a 12 day control period and during a 12 day period of treatment with human growth hormone (HGH) at the following doses: 0.0168, 0.0532, and 0.168 U/kg body weight (BW)((3/4)) per day (doses A, B, and C, respectively). In five of the six normals, dose C caused positive balances in N, P, Na, and K; doses B and A had anabolic effects in two and one normal subjects, respectively. In six of the seven Duchenne cases, dose C caused negative balances of N and K, and sometimes of P. Negative balances were produced in three of the Duchenne subjects by dose B, and in one by dose A. None of the dystrophy cases exhibited an anabolic response to any dosage of HGH tested. The release of endogenous HGH in response to insulin, after 2 days' pretreatment with diethylstilbestrol, was similar in both groups of subjects. In the course of these tests, a marked anabolic effect of diethylstilbestrol in the Duchenne patients was apparent. Therefore metabolic balance studies were repeated, in both Duchenne and normal cases, during a 12 day control period and during 12 days of treatment with diethylstilbestrol (0.106 mg/kg BW((3/4)) per day). In three of the normal children, diethylstilbestrol had no effect on the elemental balances; in two cases, a retention of Na was observed. In all seven Duchenne cases, diethylstilbestrol caused positive balances in N, P, Na, and K. Ethinyl estradiol (0.0106 mg/kg BW((3/4)) per day) produced positive N, P, Na, and K balances in all three Duchenne cases tested with this agent. The data show that exogenous HGH causes a catabolic effect in boys with Duchenne dystrophy. These patients are hyperresponsive to the anabolic effect of diethylstilbestrol. The latter phenomenon may reflect the inhibitory effect of estrogen upon the peripheral actions of these boys' endogenous HGH.

  3. Using quantum chemical modeling and calculations for evaluation of cellulose potential for estrogen micropollutants removal from water effluents.

    Ghasemi, Amin; Asgarpour Khansary, Milad; Marjani, Azam; Shirazian, Saeed


    This paper is devoted to investigate the suitability of cellulose for estrogens micropollutants removal from water effluent. For this purpose, the sorption of eight estrogens including Estradiol, Estrone, Testosterone, Progesterone, Estriol, Mestranol, Ethinylestradiol and Diethylstilbestrol were investigated. The charge density profiles and sorption curves were obtained and discussed using quantum chemical calculations where the Accelrys Materials Studio software and COSMO-SAC model were employed. The geometry optimization of compound molecule and energy minimizations was performed using the Dmol3 Module and density functional theory of generalized gradient approximate and Volsko-Wilk-Nusair functional. We found that cellulose cannot be a reliable choice of sorbent for removal of Estrone and Estradiol, but it is a poor choice of sorbent for removal of Estriol, Ethinylestradiol. Cellulose can be used for Diethylstilbestrol, Mestranol, Testosterone and Progesterone removal from estrogens containing effluents.

  4. The post-coital pills as over the counter drugs


    Post-coital pill or emergency contraceptives are birth control measures that, if taken after sexual intercourse, may prevent pregnancy. High dose of postcoital contraceptives like diethylstilbestrol & other estrogens were being used for some time without any approval by FDA. Task force on postovulatory methods of fertility regulations 1998, conducted clinical trials leading to approval of two preparations for postcoital contraception by FDA namely levonorgestrel 0.75mg & combination of 0.25 l...

  5. On the rumors about the silent spring: review of the scientific evidence linking occupational and environmental pesticide exposure to endocrine disruption health effects Rumores de uma primavera silenciosa: uma revisão das evidências científicas sobre a associação entre exposição ocupacional e ambiental a pesticidas e distúrbios endócrinos


    Occupational exposure to some pesticides, and particularly DBCP and chlordecone, may adversely affect male fertility. However, apart from the therapeutic use of diethylstilbestrol, the threat to human reproduction posed by "endocrine disrupting" environmental contaminants has not been supported by epidemiological evidence thus far. As it concerns other endocrine effects described in experimental animals, only thyroid inhibition following occupational exposure to amitrole and mancozeb has been...

  6. Determination of Dissociation Constants of Complicated Compounds by Capillary Zone Electrophoresis

    YANG, Geng-Liang; WANG, De-Xian; SUN, Su-Fang; LIU, Hai-Xing; MA, Jian-Jun


    In this work,the whole theoretical metods forthe determinaion ofpKa1 and pKa2 of complicated complicated compounds are proposed by capillary zone electrophoresis.The pka values areachieved by non-linear regression analysis by takiny into consideration the effect of activity coefficient.This is the first report on determining the dissociation constants of gastrodin,magnolol,honkiol,puercetin,curcumin,diethylstilbestrol,diehylstilbestrol,4acetamidophenol,eugenol and paeonol.

  7. Human Teratogens Update 2011: Can We Ensure Safety during Pregnancy?

    Rasmussen, Sonja A.


    Anniversaries of the identification of three human teratogens (i.e., rubella virus in 1941, thalidomide in 1961, and diethylstilbestrol in 1971) occurred in 2011. These experiences highlight the critical role that scientists with an interest in teratology play in the identification of teratogenic exposures as the basis for developing strategies for prevention of those exposures and the adverse outcomes associated with them. However, an equally important responsibility for teratologists is to ...

  8. Prostate Activated Prodrugs and Imaging Agents


    418. A Prospective, Multicenter, Randomized Phase II Trial of the Herbal Supplement, PC-SPES, and Diethylstilbestrol in Patients with Androgen...using E Merck 230-400 mesh 60H silica gel , or using a Harrison Research Inc. radial chromatotron unit. Boc-L-tyrosyl 4-(hydroxymethyl)anilide (2) To...5%, 2 ml), NaHCO3 (5%, 1 ml), then the organic extracts dried over Na2SO4. After condensation in vacuo the residue was purified by silical gel

  9. Synthesis of deuterium-labelled compounds for FOTEK project; Syntese af deuterium-maerkede forbindelser til FOeTEK projektet

    Joergensen, O.; Egsgaard, H.; Larsen, E. [Forskningscenter Risoe, Roskilde (Denmark)


    In the FoTech project there have been utilized labelled compounds of stable isotopes as internal standards. Some of these compounds are commercially available ({sup 13}C-labelled PCB congeners, {sup 13}C-labelled diethylstilbestrol for determination of anabolic steroids). Others, like D{sub 9}-clenbuterol, D{sub 3}-clenbuterol, D{sub 3}-zeramol and D{sub 3}-dimetridazol have been synthesized. General aspects of deuterium compounds labelling are considered. (EG).

  10. Using Fenton Oxidation to Simultaneously Remove Different Estrogens from Cow Manure

    Minxia Sun; Defu Xu; Yuefei Ji; Juan Liu; Wanting Ling; Shunyao Li; Mindong Chen


    The presence of estrogens in livestock excrement has raised concerns about their potential negative influence on animals and the overall food cycle. This is the first investigation to simultaneously remove estrogens, including estriol (E3), bisphenol A (BPA), diethylstilbestrol (DES), estradiol (E2), and ethinyl estradiol (EE2), from cow manure using a Fenton oxidation technique. Based on the residual concentrations and removal efficiency of estrogens, the Fenton oxidation reaction conditions...

  11. Assessment of stilbene residues in cattle through analytical control in Korca region



    Full Text Available The use of substances having hormonal action is banned in Albania. However, sometimes forbidden drugs may be added to feeds for illegal administration or treatment to cattle for promoting increased muscle development or increased water retention and thus obtain an economical benefit. Residues of these substances may remain in meat and may pose a real threat to the consumer either through exposure to the residues. On this context use of stilbens as hormonal dugs in cattle is used in illegal way. Evaluation of stilbens residues in live cattle and beef meat samples remains a common objective of food control in Albania. Assessment of stilbene residues (diethylstilbestrol, hexerol and dienstrol is carried out from 2012 to 2013 in 94 urine samples collected from cattle in region of Korca. Analytical control is performed by ELISA test as commercial product following the use instructions. Study results showed the positive results for stilbens group of substances in 8, 5% (8/ 94 of urine samples. Detection limit of ELISA test is respectively 0.15ng/ml for diethylstilbestrol, 0.25ng/ml for hexerol and 0.5ng/ml for dienstrol residues. 6 out 8 positive urine samples for stilbens residues contained diethylstilbestrol confirming as well use of hormones in cattle treatment.

  12. Development of UDP-glucuronosyltransferase activity toward digitoxigenin-monodigitoxoside in neonatal rats.

    Watkins, J B; Klaassen, C D


    Glucuronidation is low or undetectable in embryonic and early fetal tissues and changes to adult levels at rates depending on the acceptor, tissue, and species. Because other data indicate there may be a specific UDP-glucuronosyltransferase (GT) in the liver of adult rats that glucuronidates digitoxigenin-monodigitoxoside (DIG), the development of GT activity in neonatal rats toward DIG was compared with that of other acceptors. Conjugation of p-nitrophenol and 1-naphthol was higher at birth and decreased to adult levels by 20 days of age. Glucuronidation of chloramphenicol, morphine, valproic acid, and bilirubin increased from birth to adult activity by 20 days of age. Conjugation of phenolphthalein, estrone, and diethylstilbestrol was low in 1-day-old rats and higher than adult in 20-day-old animals. In contrast, glucuronidation of DIG was barely detectable (9% of adult) in 20-day-old rats. The concentration of UDP-glucuronic acid was 50% of adult levels at birth and increased to adult values by 10 days of age. Administration of 3-methylcholanthrene on days 6 to 9 after birth significantly stimulated GT activity toward 1-naphthol, p-nitrophenol, and morphine, whereas phenobarbital precociously increased conjugation of chloramphenicol, valproic acid, morphine, and diethylstilbestrol. Pregnenolone-16 alpha-carbonitrile enhanced the development of GT activity toward morphine, chloramphenicol, valproic acid, bilirubin, diethylstilbestrol, and estrone. Glucuronidation of DIG was not increased after 3-methylcholanthrene or phenobarbital, but could be induced after pregnenolone-16 alpha-carbonitrile to 7% of adult values in 10-day-old rats.(ABSTRACT TRUNCATED AT 250 WORDS)

  13. Assaying estrogenicity by quantitating the expression levels of endogenous estrogen-regulated genes.

    Jørgensen, M; Vendelbo, B; Skakkebaek, N E; Leffers, H


    Scientific evidence suggests that humans and wildlife species may experience adverse health consequences from exposure to environmental chemicals that interact with the endocrine system. Reliable short-term assays are needed to identify hormone-disrupting chemicals. In this study we demonstrate that the estrogenic activity of a chemical can be evaluated by assaying induction or repression of endogenous estrogen-regulated "marker genes" in human breast cancer MCF-7 cells. We included four marker genes in the assay--pS2, transforming growth factor beta3 (TGFbeta3), monoamine oxidase A, and [alpha]1-antichymotrypsin--and we evaluated estrogenic activity for 17beta-estradiol (E(2)), diethylstilbestrol, [alpha]-zearalanol, nonylphenol, genistein, methoxychlor, endosulphan, o,p-DDE, bisphenol A, dibutylphthalate, 4-hydroxy tamoxifen, and ICI 182.780. All four marker genes responded strongly to the three high-potency estrogens (E(2), diethylstilbestrol, and [alpha]-zearalanol), whereas the potency of the other chemicals was 10(3)- to 10(6)-fold lower than that of E(2). There were some marker gene-dependent differences in the relative potencies of the tested chemicals. TGFbeta3 was equally sensitive to the three high-potency estrogens, whereas the sensitivity to [alpha]-zearalanol was approximately 10-fold lower than the sensitivity to E(2) and diethylstilbestrol when assayed with the other three marker genes. The potency of nonylphenol was equal to that of genistein when assayed with pS2 and TGFbeta3, but 10- to 100-fold higher/lower with monoamine oxidase A and [alpha]1-antichymotrypsin, respectively. The results are in agreement with results obtained by other methods and suggest that an assay based on endogenous gene expression may offer an attractive alternative to other E-SCREEN methods.

  14. Bisphenol a exposure: human risk and health policy.

    Erler, Cheryl; Novak, Julie


    Bisphenol A (BPA) is a chemical used extensively to manufacture commonly used plastics and epoxy resin liners for food and beverage cans. BPA, with properties similar to diethylstilbestrol, has been shown to exert endocrine-disrupting effects and result in behavioral changes, altered growth, and early secondary sexual maturation. In 2008, legislation was introduced at the state and federal level to ban the use of BPA in children's products. The purpose of this article is to provide the reader with the weight of evidence, current federal regulatory stance, and proposed legislation regarding the safe use of BPA.

  15. Clear cell adenocarcinoma of the ulterine cervix in a 15 year old girl: A case report

    Choi, Seung Joon; Kim, Jee Eun; KIm, Hyung Sik; Choi, Hye Young [Dept. of Radiology, Gachon University Gil Hospital, Incheon (Korea, Republic of)


    Cervical cancer is rare in the pediatric population. In cases of cervical cancer, adenocarcinoma is predominantly reported. Clear cell adenocarcinoma (CCAC) of the uterine cervix is a very rare tumor and accounts for only 4% of all adenocarcinomas of the uterine cervix. Risk factors and pathogenesis of this disease are not exactly revealed. The intrauterine exposure to diethylstilbestrol (DES) and associated non-steroidal estrogen during pregnancy before 18 weeks is the only known risk factor. This study reports the imaging finding of primary uterine cervical tumor in a 15-year-old girl, who was finally diagnosed with CCAC, with no maternal history of DES exposure in utero.

  16. Use of radioimmunoassay procedures for the determination of sex hormones in animal tissues

    Hoffmann, B. (Institut fuer Veterinaermedizin des Bundesgesundheitsamtes (Robert von Ostertag-Institut), Berlin (Germany, F.R.))


    Radioimmunoassay methods for the determination of sex steroids and other compounds with sex hormone-like activities in various edible animal tissues and endocrine glands have been developed. Reliability of these methods, allowing quantification in a range of 10/sup -11/ M, has been adequately demonstrated. When applied to monitoring residues of anabolic sex hormones in edible tissues of veal calves, physiological baseline levels of some endogenous ''anabolic'' steroids (like testosterone, oestrogens) were established; in the case of xenobiotics residues at the scheduled time of slaughter could be quantified (trenbolone) and a regulatory method to implement the ban of diethylstilbestrol was introduced.

  17. Evaluation of Roles of Interferon Gamma Regulated Genes in Inhibition of Androgen-Independent Prostate Cancer


    NM_199170 56937 -2.6 AR PPAP2A phosphatidic acid phosphatase type 2A CR617429 8611 -2.5 EBP emopamil binding protein (sterol isomerase) CN395741 10682 -2.2...BC062755 4638 3.9 AR MYH3 myosin, heavy polypeptide 3, skeletal muscle , embryonic CK824450 4621 1.8 SPARC secreted protein, acidic , cysteine-rich...The effects of diethylstilbestrol and castration on the nucleic acid and protein metabolism of rat prostate gland. Biochem J 1976; 104: 1075-81

  18. Bedside etiology of childhood cancer. [X radiation, /sup 224/Ra

    Miller, R.W.


    The following topics are discussed: adenocarcinomas of the vagina in young women exposed prenatally to diethylstilbestrol; neuroblastoma in children exposed prenatally to hydantoin; increased frequency of leukemia in Japanese children surviving the atomic bomb; cancer in adults following administration of /sup 224/Ra during childhood; increased risk of lung cancer following environmental exposure of children to asbestos; lung cancer in adults following exposure to lead, copper, zinc, and arsenic; association of chromosomal aberrations with leukemia in children exposed to x radiation and benzene; immunosuppression in lymphoma patients; renal dysplasia in patients with Wilms' tumor; and aggregation of neoplasms in families. (HLW)

  19. Atypical McMurry Cross-Coupling Reactions Leading to a New Series of Potent Antiproliferative Compounds Bearing the Key [Ferrocenyl-Ene-Phenol] Motif

    Pascal Pigeon; Meral Görmen; Konrad Kowalski; Helge Müller-Bunz; McGlinchey, Michael J.; Siden Top; Gérard Jaouen


    In the course of the preparation of a series of ferrocenyl derivatives of diethylstilbestrol (DES), in which one of the 4-hydroxyphenyl moieties was replaced by a ferrocenyl group, the McMurry reaction of chloropropionylferrocene with a number of mono-aryl ketones unexpectedly yielded the hydroxylated ferrocenyl DES derivatives, 5a–c, in poor yields (10%–16%). These compounds showed high activity on the hormone-independent breast cancer cell line MDA-MB-231 with IC50 values ranging from 0.14 ...

  20. Acute relaxation of mouse duodenum [correction of duodenun] by estrogens. Evidence for an estrogen receptor-independent modulation of muscle excitability.

    Díaz, Mario; Ramírez, Cristina M; Marin, Raquel; Marrero-Alonso, Jorge; Gómez, Tomás; Alonso, Rafael


    17-beta-Estradiol, the stereoisomer 17-alpha-estradiol and the synthetic estrogen diethylstilbestrol (DES), all caused a rapid (verruculogen. The effects of BAY-K8644 and K(+) channel blockers were synergistic, and allowed relaxed tissues to recover spontaneous activity and basal tone. We hypothesize that the rapid non-genomic spasmolytic effect of estrogens on mouse duodenal muscle might be triggered by an estrogen-receptor-independent mechanism likely involving activation of tetraethylamonium- and 4-aminopyridine-sensitive K(+) channels and inhibition of L-type Ca2(+) channels on the smooth muscle cells.

  1. Diversification of CDK11 transcripts during chicken testis development and regression.

    Francone, Victor P; Mezquita, Jovita


    CDK11 (cyclin-dependent kinase 11, formerly known as PITSLRE) is a serine/threonine kinase that associates with the cyclin L2 regulatory partner. CDK11 catalytic activity has been associated with apoptosis, transcription, and RNA processing. Here, we identify novel chicken testis CDK11 transcripts that differ in their 5'UTR, 3'UTR, splicing of the exon 6, and polyadenylation. We have also characterized the differential expression of CDK11 in somatic tissues, during testis development and upon testicular regression by diethylstilbestrol (DES) treatment. The heterogeneity of CDK11 transcripts presented in this study suggests new possibilities for post-transcriptional regulation.

  2. Hollow fiber membrane-coated functionalized polymeric ionic liquid capsules for direct analysis of estrogens in milk samples.

    Feng, Juanjuan; Sun, Min; Bu, Yanan; Luo, Chuannan


    Protein removal process is always time-consuming for the analysis of milk samples. In this work, hollow fiber membrane-coated functionalized polymeric ionic liquid (HF-PIL) capsules were synthesized and used as solid-phase microextraction (SPME) sorbent for direct analysis of estrogens in milk samples. The functionalized PIL monolith sorbent was obtained by copolymerization between 1-(3-aminopropyl)-3-(4-vinylbenzyl)imidazolium 4-styrenesulfonate IL monomer and 1,6-di(3-vinylimidazolium) hexane bishexafluorophosphate IL-crosslinking agent. A group of four capsules were installed as SPME device, to determine four kinds of estrogens (estrone, diethylstilbestrol, hexestrol, and 17α-ethynylestradiol) in milk samples, coupled to high performance liquid chromatography. Extraction and desorption conditions were optimized to get satisfactory extraction efficiency. Good linearity was obtained in the range of 5-200 μg L(-1). The limits of detection were 1 μg L(-1) for diethylstilbestrol and 2 μg L(-1) for 17α-ethynylestradiol, estrone, and hexestrol. The present method was applied to analyze the model analytes in different milk samples. Relative recoveries were in the range of 85.5-112%. The HF-PIL SPME capsules showed satisfactory extraction efficiency and high resistance to sample matrix interference.

  3. Magnetic molecularly imprinted polymers synthesized by surface-initiated reversible addition-fragmentation chain transfer polymerization for the enrichment and determination of synthetic estrogens in aqueous solution.

    Chen, Fangfang; Zhang, Jingjing; Wang, Minjun; Kong, Jie


    Magnetic molecularly imprinted polymers have attracted significant interest because of their multifunctionality of selective recognition of target molecules and rapid magnetic response. In this contribution, magnetic molecularly imprinted polymers were synthesized via surface-initiated reversible addition addition-fragmentation chain transfer polymerization using diethylstilbestrol as the template for the enrichment of synthetic estrogens. The uniform imprinted surface layer and the magnetic property of the magnetic molecularly imprinted polymers favored a fast binding kinetics and rapid analysis of target molecules. The static and selective binding experiments demonstrated a desirable adsorption capacity and good selectivity of the magnetic molecularly imprinted polymers in comparison to magnetic non-molecularly imprinted polymers. Accordingly, a corresponding analytical method was developed in which magnetic molecularly imprinted polymers were employed as magnetic solid-phase extraction materials for the concentration and determination of four synthetic estrogens (diethylstilbestrol, hexestrol, dienestrol, and bisphenol A) in fish pond water. The recoveries of these synthetic estrogens in spiked fish pond water samples ranged from 61.2 to 99.1% with a relative standard deviation of lower than 6.3%. This study provides a versatile approach to prepare well-defined magnetic molecularly imprinted polymers sorbents for the analysis of synthetic estrogens in water solution. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Staging for vaginal cancer.

    Rajaram, Shalini; Maheshwari, Amita; Srivastava, Astha


    Vaginal cancer is a rare cancer comprising about 3% of all gynecologic cancers. Primary vaginal cancer should be carefully assigned as spread from cervix, vulva, and other metastatic tumors to vagina can occur. Although vaginal cancer traditionally occurs in older postmenopausal women, the incidence of high-risk human papillomavirus (HPV)-induced cancers is increasing in younger women. Squamous cell carcinoma is still the most common histopathologic type followed by adenocarcinoma. With decreasing use of diethylstilbestrol in pregnancy, non-diethylstilbestrol-associated cancers are described. The Federation Internationale de Gynecologie et d'Obstetrique (FIGO) staging of vaginal cancer (2009) follows the same rules as cervical cancer; it is clinically staged and allows the use of routine investigative modalities for staging. Although FIGO encourages the use of advanced imaging modalities, such as computed tomography, magnetic resonance imaging (MRI), and positron emission tomography (PET), to guide therapy, the imaging findings may not be used to change or reassign the stage. TNM staging is the pathologic staging system proposed by the American Joint Committee on Cancer, and information available from examination of the resected specimen, including pelvic and inguinal lymph nodes, may be used for staging. Copyright © 2015 Elsevier Ltd. All rights reserved.

  5. The presence of high-affinity, low-capacity estradiol-17β binding in rainbow trout scale indicates a possible endocrine route for the regulation of scale resorption

    Persson, Petra; Shrimpton, J.M.; McCormick, S.D.; Bjornsson, Bjorn Thrandur


    High-affinity, low-capacity estradiol-17β (E2) binding is present in rainbow trout scale. The Kd and Bmax of the scale E2 binding are similar to those of the liver E2 receptor (Kd is 1.6 ± 0.1 and 1.4 ± 0.1 nM, and Bmax is 9.1 ± 1.2 and 23.1 ± 2.2 fmol x mg protein-1, for scale and liver, respectively), but different from those of the high-affinity, low-capacity E2 binding in plasma (Kd is 4.0 ± 0.4 nM and Bmax is 625.4 ± 63.1 fmol x mg protein-1). The E2 binding in scale was displaced by testosterone, but not by diethylstilbestrol. Hence, the ligand binding specificity is different from that of the previously characterized liver E2 receptor, where E2 is displaced by diethylstilbestrol, but not by testosterone. The putative scale E2 receptor thus appears to bind both E2 and testosterone, and it is proposed that the increased scale resorption observed during sexual maturation in both sexes of several salmonid species may be mediated by this receptor. No high-affinity, low-capacity E2 binding could be detected in rainbow trout gill or skin.

  6. Potential role of redox cycling as a mechanism for chemical teratogenesis

    Juchau, M.R.; Fantel, A.G.; Harris, C.; Beyer, B.K.


    A survey of the literature indicates that several chemicals whose reduced metabolites are capable of undergoing redox cycling in biological systems also possess significant teratogenic properties when tested in vivo. The authors have initiated investigations to determine whether the embryotoxic effects of such chemicals could result from their redox cycling properties and whether redox cycling could be an important mechanism in chemical teratogenesis. In order to obviate the potentially confounding influences of maternal factors, the initial studies have been performed with a whole embryo culture system with redox cycling agents added directly to the culture medium. Several representative redox cycling agents including doxorubicin, paraquat, a series of nitroheterocycles, nitrosofluorene, and diethylstilbestrol (converted metabolically to redox cycling quinone/semiquinone radicals) have been investigated thus far. The nitroheterocycles which bear nitro groups with comparatively high redox potentials produced a striking, asymmetric defect involving primarily the right half of the prosencephalic and mesencephalic regions. The effect was exacerbated under conditions of low O/sub 2/ tension. Accumulated data to date strongly suggest that reduction of the nitro group is an essential feature in the embryotoxic mechanism. Quinones (doxorubicin, paraquat) and compounds metabolically converted to quinones (diethylstilbestrol) appeared to produce embryotoxic effects via mechanisms not associated with redox cycling. Nitrosofluorene embryotoxicity was markedly exacerbated by changes in both intra- and extracellular glutathione levels, but definitive dependence on a radical-mediated effect or redox cycling was not demonstrated.

  7. One-pot synthesized functionalized mesoporous silica as a reversed-phase sorbent for solid-phase extraction of endocrine disrupting compounds in milks.

    Gañán, Judith; Morante-Zarcero, Sonia; Pérez-Quintanilla, Damián; Marina, María Luisa; Sierra, Isabel


    A new procedure for the determination of 12 naturally occurring hormones and some related synthetic chemicals in milk, commonly used as growth promoters in cattle, is reported. The method is based on liquid-liquid extraction followed by solid-phase extraction (SPE) using a new one-pot synthesized ordered mesoporous silica (of the SBA-15 type) functionalized with octadecyl groups (denoted as SBA-15-C18-CO) as reversed-phase sorbent. The analytes were eluted with methanol and then submitted to HPLC with diode array detection. Under optimal conditions, the method quantification limit for the analytes ranged from 0.023 to 1.36μg/mL. The sorbent affored the extraction of estrone, 17β-estradiol, estriol, progesterone, hexestrol, diethylstilbestrol, 4-androstene-3,17-dione, ethinylestradiol, 17α-methyltestosterone, nandrolone, prednisolone and testosterone with mean recoveries ranging from 72% to 105% (except for diethylstilbestrol) with RSDextraction methods, therefore SBA-15-C18-CO mesoporous silica possess a high potential as a reversed-phase sorbent for SPE of the 12 mentioned endocrine disrupting compounds in milk samples.

  8. DNA interstrand cross-link induced by estrogens as well as their complete and synergic carcinogenesis


    The estrogens show negative activity in Ames test, but estrodiol and diethylstilbestrol in estrogens both are carcinogens based upon animal experiments and epidemiological investigation. It is concluded from the di-region theory, a mechanism conception put forward by one of the present authors, that the carcinogenesis of estrogens is switched on by the covalent cross-link between complementary DNA bases induced by them. We verified for the first time by the DNA alkaline elution method that both estrodiol and diethylstilbestrol cause covalent cross-link between DNA-protein and DNA interstrands after metabolic activation with dosage correlation, but neither the non-carcinogens cholesterol nor pyrene can lead to these sorts of cross-link in the same condition. It has been known that there is a synergetic effect between estrogen and pollution of polycyclic aromatic hydrocarbons. Although non-carcinogenic pyrene alone cannot induce cross-link, its addition with equal molar quantity to estrodiol culture causes synergically the total and DNA interstrand cross-link ratios to be respectively four and three times more than the ones in the cultivation with estrodiol only. It is shown that not only the estrodiol set off the formation of pyrene bi-radicals, but also the pyrene radicals arouse conversely the production of estrodiol bi-radicals.

  9. [Effects of Yifuning capsule on blood lipids of ovariectomized hyperlipidemia rats].

    Wang, Zhi-xia; Deng, Hong-zhu; Chen, Jian-guo; Liu, Pei-zhong


    To observe the effects of Yifuning (YEN) capsule on blood lipids of ovariectomized hyperlipidemia rats. Fifty-six female mature Sprague-Dawley rats were randomized into 7 groups: normal control group, model control group, diethylstilbestrol tablets (DT) group, Xuezhikang group, YFN high, middle and low dose groups. The ovariectomized rats were fed on high fat diet and administrated with the drugs for 3 weeks, then were killed and estimated body weight, liver index and five items of blood lipid (TC, TG, HDL-C, LDL-C, VLDL) by test kit. Enzyme (such as HP, LDL, and whole lipase) was detected too. The weight and liver index of model control group increased obviously as compared with normal group. YFN could reduce TG, TC, and LDL-C (P LDL and whole lipase (P < 0.05) on the other hand. YFN can ameliorate blood lipids of ovariectomized hyperlipidemia rats.

  10. Developmental Exposure to Endocrine Disruptors and the Obesity Epidemic

    Newbold, Retha R.; Padilla-Banks, Elizabeth; Snyder, Ryan J.; Phillips, Terry M.; Jefferson, Wendy N.


    Xenobiotic and dietary compounds with hormone-like activity can disrupt endocrine signaling pathways that play important roles during perinatal differentiation and result in alterations that are not apparent until later in life. Evidence implicates developmental exposure to environmental hormone-mimics with a growing list of health problems. Obesity is currently receiving needed attention since it has potential to overwhelm health systems worldwide with associated illnesses such as diabetes and cardiovascular disease. Here, we review the literature that proposes an association of exposure to environmental endocrine disrupting chemicals with the development of obesity. We describe an animal model of developmental exposure to diethylstilbestrol (DES), a potent perinatal endocrine disruptor with estrogenic activity, to study mechanisms involved in programming an organism for obesity. This experimental animal model provides an example of the growing scientific field termed “the developmental origins of adult disease” and suggests new targets of abnormal programming by endocrine disrupting chemicals. PMID:17321108

  11. The emergence of molecular gynecology: homeobox and Wnt genes in the female reproductive tract.

    Kitajewski, J; Sassoon, D


    Reproductive tissues respond to steroid hormones and thus are particularly vulnerable to the effects of exogenous steroid 'mimic' compounds (endocrine disrupters). One such endocrine disrupter, diethylstilbestrol (DES), is linked to gynecological cancers and changes in uterine structure that reduce or completely abrogate reproductive competence. Until recently, little was known about the identity of target genes and signaling pathways involved in pathologies linked to endocrine disrupters such as DES. We outline genetic, cellular and molecular roles for patterning genes, with emphasis on homeobox and Wnt genes. There is evidence that changes in the expression of Wnt and homeogenes underlie many of the defects induced by DES. Data obtained from murine systems will likely apply to a broad spectrum of gynecological pathologies involving abnormal cell behaviors ranging from fibroids to malignant tumors. Knowledge garnered from modern molecular genetics should lead to progress in the emerging field of molecular gynecology.

  12. Endocrine-disrupting chemicals use distinct mechanisms of action to modulate endocrine system function.

    Henley, Derek V; Korach, Kenneth S


    The term endocrine-disrupting chemicals is used to define a structurally diverse class of synthetic and natural compounds that possess the ability to alter various components of the endocrine system and potentially induce adverse health effects in exposed individuals and populations. Research on these compounds has revealed that they use a variety of both nuclear receptor-mediated and non-receptor-mediated mechanisms to modulate different components of the endocrine system. This review will describe in vitro and in vivo studies that highlight the spectrum of unique mechanisms of action and biological effects of four endocrine-disrupting chemicals--diethylstilbestrol, genistein, di(n-butyl)phthalate, and methoxyacetic acid--to illustrate the diverse and complex nature of this class of compounds.

  13. Does cancer start in the womb? altered mammary gland development and predisposition to breast cancer due to in utero exposure to endocrine disruptors.

    Soto, Ana M; Brisken, Cathrin; Schaeberle, Cheryl; Sonnenschein, Carlos


    We are now witnessing a resurgence of theories of development and carcinogenesis in which the environment is again being accepted as a major player in phenotype determination. Perturbations in the fetal environment predispose an individual to disease that only becomes apparent in adulthood. For example, gestational exposure to diethylstilbestrol resulted in clear cell carcinoma of the vagina and breast cancer. In this review the effects of the endocrine disruptor bisphenol-A (BPA) on mammary development and tumorigenesis in rodents is used as a paradigmatic example of how altered prenatal mammary development may lead to breast cancer in humans who are also widely exposed to it through plastic goods, food and drink packaging, and thermal paper receipts. Changes in the stroma and its extracellular matrix led to altered ductal morphogenesis. Additionally, gestational and lactational exposure to BPA increased the sensitivity of rats and mice to mammotropic hormones during puberty and beyond, thus suggesting a plausible explanation for the increased incidence of breast cancer.

  14. Effects of soybean isoflavones on reproductive parameters in Chinese mini-pig boars

    Yuan Xiao-xue


    Full Text Available Abstract Background Soybean isoflavones are structurally similar to mammalian estrogens and therefore may act as estrogen agonists or antagonists. However, it has not been determined if they have any negative effects on reproductive parameters in male livestock. Therefore, the objective of this study was to evaluate the effects of soybean isoflavones on male reproduction using Chinese mini-pig boars as a model. Fifty Xiang boars were randomly divided into five groups and fed diets containing 0, 125, 250, or 500 ppm soybean isoflavones or 0.5 ppm diethylstilbestrol for 60 days. Results Dietary supplementation with 250 ppm of soy isoflavones markedly increased the testis index (P P P P P P P P P P P Conclusions The results of this study indicate that consumption of soy isoflavones at dietary levels up to 250 ppm did not adversely affect reproductive parameters in Chinese mini-pig boars whereas higher levels of soy isoflavones may adversely affect male reproduction.

  15. Clear cell carcinoma of the female genital tract (not everything is as clear as it seems).

    Offman, Saul L; Longacre, Teri A


    Clear cell carcinoma has a storied history in the female genital tract. From the initial designation of ovarian clear cell adenocarcinoma as "mesonephroma" to the linkage between vaginal clear cell carcinoma and diethylstilbestrol exposure in utero, gynecologic tract clear cell tumors have puzzled investigators, posed therapeutic dilemmas for oncologists, and otherwise presented major differential diagnostic challenges for pathologists. One of the most common errors in gynecologic pathology is misdiagnosis of clear cell carcinoma, on both frozen section and permanent section. Given the poor response to platinum-based chemotherapy for advanced-stage disease and increased risk of thromboembolism, accurate diagnosis of clear cell carcinoma is important in the female genital tract. This review (1) presents the clinical and pathologic features of female genital tract clear cell carcinomas; (2) highlights recent molecular developments; (3) identifies areas of potential diagnostic confusion; and (4) presents solutions for these diagnostic problems where they exist.

  16. Selection of chemotherapy for metastatic mammary cancer by effect on cesium-131 uptake.

    Ferguson, D J; Harper, P V


    Cesium-131 was administered intravenously to 39 patients with superficial metastases of mammary carcinoma and the concentration in tumor was compared with that in normal tissue by application of a detector in vivo, before and after 1 to 5 days of chemotherapy with cyclophosphamide (CP), 5-fluorouracil (FU), or diethylstilbestrol. A change of the cesium concentration ratio (tumor/normal tissue) greater than 15% after brief treatment correctly predicted the therapeutic effect after 1 to 39 months on the tumors that were tested in 30 of 33 tests. No reliable correlation could be made in the remaining 21 tests in which the change of ratio was less than 15%. The concentration of cesium-131 in the skin, fat, and skeletal muscle of mice was not appreciably altered by treatment for 5 days with CP or FU.

  17. Voltammetric Determination of Estrogens Based on the Enhancement Effect of Surfactant at Carbon Paste Electrode


    Highly sensitive voltammetric method for the determination of estrogens, based on the enhancement effect of cetyltrimethylammonium bromide (CTAB) has been described. In the presence of CTAB, the oxidation peak currents of estrogens (estradiol, estrone, estriol, estradiol valerate and diethylstilbestrol) at the carbon paste electrode (CPE) increased significantly after open-circuit accumulation. The peak current was proportional to the concentration of estradiol over the range from 5×10-9 to 2.5×10-6 mol\\5L-1. The detection limit was 8×10-10 mol\\5L-1 at 6 min of accumulation. The total amounts of estrogens in the blood serums were determined and the average recovery was 104.92%. Under the conditions used, the electrode process of estradiol was examined and the mechanism for peak current enhancement was also discussed.

  18. 激素类药品的拉曼光谱研究%Raman Spectroscopy of Hormonal Drugs

    修梓侨; 梅雪峰; 孙秀平


    应用LRS-Ⅱ型激光拉曼光谱仪,对地塞米松(C22H29FO5),甲状腺素(C15H11O4I4N),乙烯雌酚(C18H20O2)这三种激素类药品进行拉曼光谱检测,确定其拉曼光谱位置,理论分析这几种物质的内部结构和拉曼光谱关系.同时利用高斯软件对这三种激素类药品进行分析比对模拟计算.结果表明,利用拉曼光谱法可以快速准确地检测出地塞米松、甲状腺素、乙烯雌酚这三种激素类药品,并且对其拉曼光谱特征峰归属进行了指认.%LRS-Ⅱ laser Raman spectrograph was applied to detect the Raman characterization of dexamethasone (C22H29FO5),thyroxine(C15H11O4I4N) and diethylstilbestrol (C18H20O2) and to determine their positions of Raman spectroscopy, also to analysis the relationship between the internal structure and Raman spectra of these three pharma-ceuticals. Moreover, the analysis of simulation calculation of these three pharmaceuticals was carried out by using Gaussian software. Results has shown that rapid and accurate qualitative detection of these three pharmaceuticals of dexamethasone,thyroxine and diethylstilbestrol was achieved by Raman spectrograph,also the identification of the char-acteristic peaks of Raman spectra was obtained.

  19. Activation of cellular oncogenes by chemical carcinogens in Syrian hamster embryo fibroblasts

    Ebert, R.; Reiss, E.; Roellich, G.; Schiffmann, D. (Univ. of Wuerzburg (West Germany)); Barrett, J.C.; Wiseman, R.W. (National Institute of Environmental Health Sciences, Research Triangle Park, NC (USA)); Pechan, R.


    Carcinogen-induced point mutations resulting in activation of ras oncogenes have been demonstrated in various experimental systems such as skin carcinogenesis, mammary, and liver carcinogenesis. In many cases, the data support the conclusion that these point mutations are critical changes in the initiation of these tumors. The Syrian hamster embryo (SHE) cell transformation model system has been widely used to study the multistep process of chemically induced neoplastic transformation. Recent data suggest that activation of the Ha-ras gene via point mutation is one of the crucial events in the transformation of these cells. The authors have now cloned the c-Ha-ras proto-oncogene from SHE cDNA-libraries, and we have performed polymerase chain reaction and direct sequencing to analyze tumor cell lines induced by different chemical carcinogens for the presence of point mutations. No changes were detectable at codons 12, 13, 59, 61, and 117 or adjacent regions in tumor cell lines induced by diethylstilbestrol, asbestos, benzo(a)pyrene, trenbolone, or aflatoxin B{sub 1}. Thus, it is not known whether point mutations in the Ha-ras proto-oncogene are essential for the acquisition of the neoplastic phenotype of SHE cells. Activation of other oncogenes or inactivation of tumor suppressor genes may be responsible for the neoplastic progression of these cells. However, in SHE cells neoplastically transformed by diethylstilbestrol or trenbolone, a significant elevation of the c-Ha-ras expression was observed. Enhanced expression of c-myc was detected in SHE cells transformed by benzo(a)pyrene or trenbolone.

  20. Competitive advantage and tolerance of selected shochu yeast in barley shochu mash.

    Takashita, Hideharu; Fujihara, Emi; Furutera, Mihoko; Kajiwara, Yasuhiro; Shimoda, Masahiko; Matsuoka, Masayoshi; Ogawa, Takahira; Kawamoto, Seiji; Ono, Kazuhisa


    A shochu yeast strain, Saccharomyces cerevisiae BAW-6, was previously isolated from Kagoshima yeast strain Ko, and has since been utilized in shochu production. The BAW-6 strain carries pho3/pho3 homozygous genes in contrast to the heterozygous PHO3/pho3 genes in the parental Ko strain. However, absence of the PHO3 gene per se cannot explain the fermentation superiority of BAW-6. Here, we demonstrate the growth advantage of the BAW-6 strain over the Ko strain by competitive cultivation in barley shochu preparation, where alcohol yield and nihonshudo of the former strain were higher than those of the latter strain. In addition, the maximum growth rate of BAW-6 was less affected than that of Ko by high Brix values of barley koji medium, suggesting that BAW-6 is less sensitive to growth inhibitory compounds derived from barley or barley koji. The tolerance of BAW-6 to growth inhibitory compounds, cerulenin and diethylstilbestrol (an H⁺-ATPase inhibitor), was also higher than that of other yeast strains. Consistent with BAW-6's tolerance to diethylstilbestrol in the presence of 8% ethanol (pH 4.5), H⁺-ATPase activity, but not transcription of its gene, was higher in BAW-6 than in Ko. We conclude that the BAW-6 strain is associated with certain gene alterations other than PHO3, such that it can maintain cellular ion homeostasis under conditions of ethanol stress during the latter phase of fermentation. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  1. Luteinizing hormone receptor (lhcgr) as a marker gene for characterizing estrogenic endocrine-disrupting chemicals in zebrafish ovarian follicle cells.

    Liu, Ka-Cheuk; Wu, Rudolf S S; Ge, Wei


    The adverse effects of endocrine-disrupting chemicals (EDCs) have been well documented; however, the action mechanisms of many EDCs remain elusive and controversial. Furthermore, the highly diversified chemical structures and low environmental concentrations of EDCs present a major challenge to their chemical detection. Clearly, there is an urgent need for simple and reliable bioassays to detect EDCs in the environment and unravel their action mechanisms. We have recently identified luteinizing hormone receptor (lhcgr) as a robust estradiol (E2)-responsive gene in cultured zebrafish ovarian follicle cells. The expression of lhcgr exhibited a distinct biphasic response to E2 over a 24-h time-course treatment, making this a unique system for characterizing estrogenic EDCs. This study was undertaken to validate this platform by testing a wide range of EDCs, including 17α-ethinylestradiol (EE2), diethylstilbestrol (DES), bisphenol A (BPA), genistein (GEN), 1,1,1-trichloro-2-(2-chlorophenyl)-2-(4-chlorophenyl)ethane (o,p'-DDT), vinclozolin (VIN), bis(2-ethylhexyl) phthalate (DEHP), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), and 2,2',4,4'-tetrabromodiphenyl ether (BDE-47). Diethylstilbestrol (DES), EE2 and o,p'-DDT mimicked E2 and induced a biphasic expression of lhcgr while BPA and GEN stimulated a monophasic expression in the 24-h time-course. In contrast, BDE-47, DEHP and VIN had no effect, whereas TCDD decreased lhcgr expression. Dose-response experiment showed that E2, EE2 and DES had the highest potency, which was followed by GEN, BPA and o,p'-DDT. The effects of estrogenic EDCs were further confirmed by their potentiation of hCG-induced activin βA2 subunit (inhbab) expression. In conclusion, the present study showed that the expression of lhcgr in cultured zebrafish follicle cells and its biphasic response to estrogens provide a unique in vitro platform for screening and categorizing estrogenic substances and deciphering their action mechanisms.

  2. Characterization and application of oviductal epithelial cells in vitro in Gallus domesticus.

    Jung, Jin Gyoung; Park, Tae Sub; Kim, Jin Nam; Han, Beom Ku; Lee, Seon Duk; Song, Gwonhwa; Han, Jae Yong


    Chicken oviductal epithelium produces large quantities of egg white protein in daily cycles. In this study, we cultured and characterized oviductal epithelial cells (OECs) from juvenile (10-wk-old) chickens and from actively laying (30-wk-old) hens. The juvenile OECs were maintained over passage 25 and were positive for toluidine blue, lectin-ConA, HPA, UEA-1, WFA, WGA, anti-OVA, anti-ESR1, and anti-PGR, whereas the adult OECs were cultured over passage 6 and were positive for toluidine blue, periodic acid-Schiff, lectin-ConA, WFA, WGA, anti-OVA, anti-ESR1, and anti-PGR. To investigate the optimal concentration of steroid hormones for inducing egg white protein genes in vitro, we examined the effects of estrogen, diethylstilbestrol, progesterone, and corticosterone on OECs. Results showed that oviduct-specific levels of avidin, ovalbumin, ovomucin, lysozyme, ESR1, and PGR gene expression were significantly elevated in steroid hormone-treated OECs compared with those of untreated cells (P < 0.05). Ovalbumin protein was also secreted into culture medium from hormone-treated OECs. In addition, to examine the application of OECs for avian transgenesis, we introduced human thrombopoietin (THPO)-expressing lentiviral vector controlled by a 3.5-kb ovalbumin promoter into cultured OECs, and THPO expression was significantly induced with diethylstilbestrol or progesterone in juvenile OECs (P < 0.05) and in adult OECs (P < 0.05). In conclusion, these data demonstrate the potential of cultured OECs as a model system for providing a better understanding of the regulation of gene expression and for the production of an avian transgenic bioreactor.

  3. Depletion of hepatic uridine diphosphoglucuronic acid decreases the biliary excretion of drugs.

    Gregus, Z; Watkins, J B; Thompson, T N; Klaassen, C D


    Hepatic levels of uridine diphosphoglucuronic acid (UDPGA) in rats decreased substantially (greater than 80%) 40 min after galactosamine (GAL) (600 mg/kg i.p.) or after 1 hr of diethyl ether (DE) narcosis. Biliary excretion of several cholephils requiring glucuronidation before excretion was reduced by GAL 76, 62, 92, 90 and 97% for bilirubin, diethylstilbestrol, iopanoic acid, phenolphthalein and valproic acid, respectively. GAL treatment caused delayed plasma clearances of the parent compounds and reductions in plasma concentrations and biliary excretions of glucuronide conjugates. The degree of this reduction was related to the maximal excretion rate of the individual compounds. For phenolphthalein glucuronide and phenol-3,6-dibromphthalein disulfonate, which do not undergo conjugation, GAL had no effect on their biliary excretion. DE-induced UDPGA depletion had no effect on phenolphthalein glucuronide excretion but reduced that of phenol-3,6-dibromphthalein disulfonate 25%. DE did not affect the plasma elimination or biliary secretion of phenolphthalein. Of the other cholephils requiring conjugation, DE reduced the excretion of bilirubin, diethylstilbestrol, iopanoic acid and valproic acid by 41, 29, 76 and 28%, respectively. DE decreased the plasma elimination of the parent compounds and the appearance of the conjugates in both plasma and bile. Reduction of glucuronide excretion into bile was less pronounced at higher doses of the cholephilic anions. Neither treatment reduced in vitro hepatic UDP-glucuronosyltransferase activity toward these substrates or substantially altered extrahepatic UDPGA concentrations. Thus, both GAL and DE decreased UDPGA to similar concentrations, but the biliary excretion of compounds requiring glucuronidation before secretion was depressed to a greater extent by GAL.

  4. Investigating the in Vitro Thermal Stability and Conformational Flexibility of Estrogen Receptors as Potential Key Factors of Their in Vivo Activity.

    Le Grand, Adélaïde; André-Leroux, Gwenaëlle; Marteil, Gaëlle; Duval, Hélène; Sire, Olivier; Le Tilly, Véronique


    Among hormone-inducible transcription factors, estrogen receptors (ERs) play important roles in tissue growth and differentiation, via either direct or indirect binding, in the nucleus, to specific DNA targets called estrogen responsive elements (EREs), or through nongenomic pathways. In humans, two estrogen receptor isoforms (hERs), designated hERα and hERβ, have been identified. These two hERs, encoded by genes located on distinct chromosomes, exhibit divergent tissue-specific functions and different subcellular distributions depending on their binding status, free or complexed to their cognate ligands. Because it is hypothesized that such distinct behaviors may arise from various conformational stabilities and flexibilities, the effect of salt concentration and temperature was studied on the free and estrogen-activated hERα and hERβ. Our results show that the conformational stability of hERβ is weakly modulated by salt concentration as opposed to hERα. In addition, we show that the estrogen-bound hERs exhibit a more constrained structure than the unliganded ones and that their conformational flexibility is more affected by diethylstilbestrol binding than that of estradiol, 4-hydroxytamoxifen, or raloxifen. In line with these results, conformational analysis and computational docking were performed on hERα and hERβ, which confer molecular support of a diethylstilbestrol-induced restrained flexibility as compared to other ligands. We found that Trp383 in hERα and Trp335 in hERβ can closely interact with the NR-box motif of the H12 helix and act as a gatekeeper of the agonist-bound versus antagonist-bound conformations. Altogether, our study contributes to an improved knowledge of the diverse physicochemical properties of full-length hERs, which will help in our understanding of their distinct cellular roles in various cellular contexts.

  5. Experimental study on curative effect of tea camellia seed oil on postmenopausal osteoporosis%山茶籽油对绝经后骨质疏松症治疗作用的实验研究

    李海; 解继胜; 陈建海; 王金花; 黄海玲; 黎飚


    Objective: To observe the effect of tea camellia seed oil on bone tissue and bone metabolism of ovariectomized rats, provide an experimental basis for preventing and treating postmenopausal osteoporosis with tea camellia seed oil. Methods: A total of 80 six - month healthy female SD rats were divided into four groups, 20 rats in each group: control group, ovariectomized model group, ovariectomized model plus diethylstilbestrol group, and ovariectomized model plus tea camellia seed oil group. The rats in ovariectomized model plus diethylstilbestrol group were treated with subcutaneous injection of diethylstilbestrol for successive 12 weeks, the therapeutic dose was 300 μg/kg, twice a week; the rats in ovariectomized model plus tea camellia seed oil group were treated with pouring tea camellia seed oil into stomach for successive 12 weeks, the therapeutic dose was 5 ml/kg, once a day. The rats in the four groups were anesthetized at 12 weeks, then the blood samples of left heart were obtained to detect the levels of serum estradiol, progesterone, follicle - stimulating hormone (FSH) , luteinizing hormone (LH) , calcium, phosphorus, and alkaline phosphatase; then the rats were killed, proximal femoral sections were prepared to observe the structure of bone tissue; all the data were analyzed statistically. Results; Compared with control group, the levels of serum calcium, phosphorus, and estradiol in ovariectomized model group decreased significantly (P <0. 01 ) , while the levels of alkaline phosphatase and FSH increased significantly (P<0. 01 ) , bone sections showed obvious pathological changes of osteoporosis; compared with ovariectomized model group, the levels of serum calcium, phosphorus, and estradiol in ovariectomized model plus diethylstilbestrol group and ovariectomized model plus tea camellia seed oil group increased significantly ( P < 0. 01 ) , while the levels of alkaline phosphatase and FSH decreased significantly (P < 0.01 ) , bone sections showed

  6. 卵巢移植对去势大鼠海马神经元形态及BDNF表达的影响%Ovarian transplantation on playing morphological changes and BDNF expression in hippocampal neurons of castrated rat

    孔斌; 刘芳; 张文华; 黑常春; 张焱; 王燕蓉; 赵承军


    Objective: To observe the impact of ovarian transplantation on playing morphological changes and BDNF expression in rat hippocampal neurons, evaluation of ovarian transplantation on the protective effect in hippocampal neurons. Methods; The model groups were established by female SD rats were used for ovariectomy ( OVX) . One intervention group was transplanted the ovaries of SD rat within 3 days under the kidney capsule. Another group was injected diethyl-stilbestro( 1 ing/kg) intraperiteneally every other day. To observe hippocampal neuronal morphology by the electron microscopy and expression of BDNF by lmmunohistochemical method after the ovarian transplantation of the 4, 7, 14 and 28 days. Results: With the migration time, the serum estrogen and progesterone levels have gradually increased in ovarian transplantation group, while only estrogen have increased in diethylstilbestrol group. Morphology of hippocampal neurons in each group gradually improved, the ovarian transplantation group was significantly increased compared with diethylstilbestrol group, especially in 14 d and 28 d. Expression of BDNF in each group have all increased at every regions in the hippocampus, and the changes of transplantation group is remarkable in CA3 area in 14 d and 28 d ( P < 0.05). Conclusion: Ovarian transplantation can restore ovarian endocrine (unction, improve the morphology of hippocampal neurons andpromote the expression of BDNF. The neuroprotective role in the transplantation group was better than diethylstilbestrol group.%目的:观察卵巢移植后大鼠海马神经元形态变化及BDNF的表达,评价卵巢移植对海马神经元的保护效果.方法:制作去势SD大鼠模型,卵巢移植干预组在其肾被膜下移植出生3d内的SD乳鼠卵巢,外源性激素干预组隔天给予腹腔注射乙烯雌酚1mg/kg.分别于干预后的4、7、14和28d,电镜观察海马神经元形态及免疫组化观察BDNF的表达情况.结果:卵巢移植后随着移植时

  7. Simultaneous Determination of Residues of Five Sex Hormones in Chicken Muscle by LC-MS/MS%高效液相色谱-串联质谱法同时测定鸡肉中5种性激素残留量

    别小琳; 贺亚玲


    目的 建立同时检测鸡肉中睾酮、孕酮、去甲雄三烯醇酮、氟氢甲睾酮、己烯雌酚5种性激素残留量的方法.方法 鸡肉经前处理后,用高效液相色谱-串联质谱法(LC-MS/MS)检测5种性激素,内标法定量.结果 方法的检出限为0.01~0.06μg/kg;加标回收率为81.9%~112.6%;测定精密度均小于3.0%;睾酮、孕酮、去甲雄三烯醇酮、己烯雌酚在1ng/ml~40 ng/ml范围内、氟氢甲睾酮在2 ng/m1~80 ng/ml范围内具有良好的线性.结论 该方法灵敏度高,杂质干扰小,特异性强,是测定鸡肉中性激素残留的理想方法.%Objective To establish a method for determination of residues of testosterone, progesterone, trenbolo-ne, fluoxymesterone and diethylstilbestrol. Methods After pre - treated, the samples were detected by LC - MS/MS and quantified by internal standard method. Results Limits of detection (LOD) for five sex hormones were between 0. 01 and 0.06ug/kg, and the spike recovery rates were from 81. 9% to 112. 6%. Within the range of lng/ml to 40ng/ml, good linearity of testosterone, progesterone, trenbolone and diethylstilbestrol was presented; within the range of 2ng/ml to 80ng/ml, good linearity of fluoxymesterone was presented. Conclusion This mediod is sensitive, little interfered by impurities and of specificity for the determination of residues of sex hormones in chicken muscle.

  8. Effects of orexins A and B on expression of orexin receptors and progesterone release in luteal and granulosa ovarian cells.

    Cataldi, Natalia I; Lux-Lantos, Victoria A R; Libertun, Carlos


    Orexin-A and orexin-B are neuropeptides controlling sleep-wakefulness, feeding and neuroendocrine functions via their G protein-coupled receptors, orexin-1R and orexin-2R. They are synthesized in the lateral hypothalamus and project throughout the brain. Orexins and orexin receptors have also been described outside the brain. Previously we demonstrated the presence of both receptors in the ovary, their increased expression during proestrous afternoon and the dependence on the gonadotropins. Here we studied the effects of orexins on the mRNA expression of both receptors, by quantitative real-time PCR, on luteal cells from superovulated rat ovaries and granulosa cells from diethylstilbestrol-treated rat ovaries. Effects on progesterone secretion were also measured. In luteal cells, 1 nM of either orexin-A or orexin-B decreased progesterone secretion. Orexin-A treatment increased expression of both orexin-1R and orexin-2R mRNA. The effect on orexin-1R mRNA expression was abolished by an orexin-1R selective receptor antagonist SB-334867 and the effect on orexin-2R mRNA expression was abolished by a selective orexin-2R antagonist JNJ-10397049. Orexin-B did not modify orexin-1R mRNA expression, but increased orexin-2R mRNA expression. The effect of orexin-B on orexin-2R was abolished by a selective orexin-2R antagonist. Neither the expression of orexin receptors nor progesterone secretions by granulosa cells were affected by orexins. FSH, as positive control, increased both steroid hormones secretion, but did not induce the expression of OX receptors in granulosa cells isolated from late preantral/early antral follicles. Finally in ovaries obtained immediately after sacrifice, the expression of orexin-1R and orexin-2R was higher in superovulated rat ovaries compared to control or diethylstilbestrol treated rat ovaries. A selective presence and function of both orexinergic receptors in luteal and granulosa cells is described, suggesting that the orexinergic system may

  9. Involvement of estrogen receptors in the resveratrol-mediated increase in dopamine transporter in human dopaminergic neurons and in striatum of female mice.

    Di Liberto, Valentina; Mäkelä, Johanna; Korhonen, Laura; Olivieri, Melania; Tselykh, Timofey; Mälkiä, Annika; Do Thi, Hai; Belluardo, Natale; Lindholm, Dan; Mudò, Giuseppa


    Treatment with resveratrol (RSV) has been shown to protect vulnerable neurons after various brain injuries and in neurodegenerative diseases. The mechanisms for the effects of RSV in brain are not fully understood, but RSV may affect the expression of various gene products. RSV is structurally related to the synthetic estrogen, diethylstilbestrol so the effects of RSV may be gender-specific. Here we studied the role of RSV in the regulation of dopamine transporter (DAT) in the striatum using male and female mice. The basic levels of DAT in the striatum showed no sex difference, but the levels increased significantly by RSV (20 mg/kg i.p.) in female but not in male mice. Pretreatment of mice with the selective estrogen receptor (ER), ERα- and ERβ antagonist ICI 182,780, led to a complete block of RSV effect on DAT protein levels, suggesting that ERs are involved in the up-regulation of DAT by RSV. Similar data was also obtained in culture using human MESC2.10 and mouse SN4741 dopaminergic cells after treatment with RSV. Data further showed that RSV specifically induced gene transcription of DAT in the dopaminergic cells. These results show that estrogen receptors are involved in the up-regulation of DAT by RSV in the dopaminergic neurons, demonstrating a sex-dependent effect of RSV in the brain that may be of clinical importance. This article is part of a Special Issue entitled 'Post-Traumatic Stress Disorder'.

  10. A transgenic mouse model expressing an ERα folding biosensor reveals the effects of Bisphenol A on estrogen receptor signaling

    Sekar, Thillai V.; Foygel, Kira; Massoud, Tarik F.; Gambhir, Sanjiv S.; Paulmurugan, Ramasamy


    Estrogen receptor-α (ERα) plays an important role in normal and abnormal physiology of the human reproductive system by interacting with the endogenous ligand estradiol (E2). However, other ligands, either analogous or dissimilar to E2, also bind to ERα. This may create unintentional activation of ER signaling in reproductive tissues that can lead to cancer development. We developed a transgenic mouse model that constitutively expresses a firefly luciferase (FLuc) split reporter complementation biosensor (NFLuc-ER-LBDG521T-CFLuc) to simultaneously evaluate the dynamics and potency of ligands that bind to ERα. We first validated this model using various ER ligands, including Raloxifene, Diethylstilbestrol, E2, and 4-hydroxytamoxifen, by employing FLuc-based optical bioluminescence imaging of living mice. We then used the model to investigate the carcinogenic property of Bisphenol A (BPA), an environmental estrogen, by long-term exposure at full and half environmental doses. We showed significant carcinogenic effects on female animals while revealing activated downstream ER signaling as measured by bioluminescence imaging. BPA induced tumor-like outgrowths in female transgenic mice, histopathologically confirmed to be neoplastic and epithelial in origin. This transgenic mouse model expressing an ERα folding-biosensor is useful in evaluation of estrogenic ligands and their downstream effects, and in studying environmental estrogen induced carcinogenesis in vivo. PMID:27721470

  11. Vortex-assisted hollow fibre liquid-phase microextraction technique combined with high performance liquid chromatography-diode array detection for the determination of oestrogens in milk samples.

    Wang, Peijin; Xiao, Yu; Liu, Wenjun; Wang, Juan; Yang, Yaling


    A rapid, simple, sensitive and environmentally friendly method has been developed for the determination of three oestrogens (17β-estradiol (17β-E2), estrone (E1), and diethylstilbestrol (DES)) in milk samples by using vortex-assisted hollow fibre liquid-phase microextraction (VA-HF-LPME) and high performance liquid chromatography. Method is based on the microextraction of oestrogens from sample solution into 15 μL of nonanoic acid as extracting agent, which is placed inside the hollow fibre followed by vortex-mixing. Vortex provided effective and mild mixing of sample solution and increased the contact between analytes and boundary layers of the hollow fibre, thereby enhancing mass transfer rate and leading to high recovery of target analytes. The extraction equilibrium is achieved within 2 min. Parameters influencing the recovery were investigated and optimized. The proposed technique provided good linearity (>0.9984), repeatability (RSD = 2.56-4.38), low limits of detection (0.06-0.17 ng mL(-1)), and high enrichment factor (330).

  12. Characterization and partial purification of a proton translocating ATPase from corn coleoptile tonoplasts

    Mandala, S.M.


    ATP-dependent proton translocating activity in microsomal membranes from corn coleoptiles was characterized. Proton pumping activity, detected by either /sup 14/C-methylamine uptake or quinacrine fluorescence quenching, had a broad optimum at pH 7.5, and was substrate specific for MgATP. N,N'-dicyclohexylcarbodiimide, diethylstilbestrol, 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole, and protonophores were found to inhibit proton transport, while vanadate and oligomycin had little effect. Proton pumping activity was stimulated 10 fold with Cl/sup -/ but was almost completely inhibited by 50 mM, KNO/sub 3/. Uptake studies with /sup 36/Cl/sup -/ indicated the Cl/sup -/ was transported into the microsomal vesicles in response to the pH gradient. ATP-stimulated proton pumping activity was localized on linear density gradients. On sucrose gradients, the activity cosedimented with the marker for endoplasmic reticulum at 1.11 g/cc. Sucrose gradients prepared in the presence of MgCl/sub 2/ were used to shift the ER marker to a heavier density, away from proton pumping activity. Linear dextran gradients also resulted in a clear separation of ATP-stimulated methylamine, thiocyanate, and /sup 36/Cl/sup -/ uptake, from markers for ER, Golgi, mitochondria, and plasma membranes. The tonoplast ATPase was solubilized with octylglucoside and partially purified on linear sucrose gradients. The specific activity of the KNO/sub 3/-sensitive ATPase increased 30-fold during purification.

  13. Environmental Estrogens and Obesity

    Newbold, Retha R.; Padilla-Banks, Elizabeth; Jefferson, Wendy N.


    Many chemicals in the environment, in particular those with estrogenic activity, can disrupt the programming of endocrine signaling pathways that are established during development and result in adverse consequences that may not be apparent until much later in life. Most recently, obesity and diabetes join the growing list of adverse consequences that have been associated with developmental exposure to environmental estrogens during critical stages of differentiation. These diseases are quickly becoming significant public health issues and are fast reaching epidemic proportions worldwide. In this review, we summarize the literature from experimental animal studies documenting an association of environmental estrogens and the development of obesity, and further describe an animal model of exposure to diethylstilbestrol (DES) that has proven useful in studying mechanisms involved in abnormal programming of various differentiating estrogen- target tissues. Other examples of environmental estrogens including the phytoestrogen genistein and the environmental contaminant Bisphenol A are also discussed. Together, these data suggest new targets (i.e., adipocyte differentiation and molecular mechanisms involved in weight homeostasis) for abnormal programming by estrogenic chemicals, and provide evidence that support the scientific hypothesis termed “the developmental origins of adult disease”. The proposal of an association of environmental estrogens with obesity and diabetes expands the focus on the diseases from intervention/treatment to include prevention/avoidance of chemical modifiers especially during critical windows of development. PMID:19433252

  14. Molecular imprinted opal closest-packing photonic crystals for the detection of trace 17β-estradiol in aqueous solution.

    Sai, Na; Wu, Yuntang; Sun, Zhong; Huang, Guowei; Gao, Zhixian


    A novel opal closest-packing (OCP) photonic crystal (PC) was prepared by the introduction of molecular imprinting technique into the OCP PC. This molecular imprinted (MI)-OCP PC was fabricated via a vertical convective self-assembly method using 17β-estradiol (E2) as template molecules for monitoring E2 in aqueous solution. Morphology characterization showed that the MI-OCP PC possessed a highly ordered three-dimensional (3D) periodically-ordered structure, showing the desired structural color. The proposed PC material displayed a reduced reflection intensity when detecting E2 in water environment, because the molecular imprinting recognition events make the optical characteristics of PC change. The Bragg diffraction intensity decreased by 19.864 a.u. with the increase of E2 concentration from 1.5 ng mL(-1) to 364.5 ng mL(-1) within 6 min, whereas there were no obvious peak intensity changes for estriol, estrone, cholesterol, testosterone and diethylstilbestrol, indicating that the MI-OCP PC had selective and rapid response for E2 molecules. The adsorption results showed that the OCP structure and homogeneous layers were created in the MI-OCP PC with higher adsorption capacity. Thus, it was learned the MI-OCP PC is a simple prepared, sensitive, selective, and easy operative material, which shows promising use in routine supervision for residue detection in food and environment.

  15. Development and characterization of a nanodendritic silver-based solid-phase extraction sorbent for selective enrichment of endocrine-disrupting chemicals in water and milk samples.

    Gao, Yuanji; Xia, Bing; Liu, Jie; Ji, Baocheng; Ma, Fengwei; Ding, Lisheng; Li, Bangjing; Zhou, Yan


    In this study, 4-[4-phenylazo-phenoxy] butyl-1-thiol (AzSH) functionalized nanodendritic silver (AzS@AgNDs) materials were prepared as a solid-phase extraction (SPE) sorbent for the selective extraction of estrogens. AzS@AgNDs possess an extremely large surface-to-volume ratio and a small average particle size. The performance of the material was evaluated by selective enrichment of hexestrol, diethylstilbestrol, dienestrol and bisphenol A in water and milk samples followed by rapid ultra-performance liquid chromatography-electrospray ionization mass spectrometry (UPLC-ESI-MS) analyses. The results exhibited that AzS@AgNDs had excellent adsorption capability for the targeted estrogens. The limits of detection of the four estrogens ranged from 0.1 to 5.0 pg/mL. The recoveries of the estrogens spiked into tap water were over the range of 83.6-105.3% with relative standard deviations of 2.8-6.0%. The results indicated the capability of this method for the rapid determination of estrogens in milk and other environmental water samples. In addition, this method would be useful for the determination of human exposure and health risk assessments trace level of endocrine-disrupting compounds (EDCs) in drinking water.

  16. In Utero Exposure to Low Doses of Bisphenol A Lead to Long-term Deleterious Effects in the Vagina

    G. Schönfelder


    Full Text Available The origins of the “endocrine disrupter hypothesis” may be traced to reports on adolescent daughters born to women who had taken the highly potent synthetic estrogen, diethylstilbestrol, while pregnant, and who developed a rare form of vaginal cancer and adenocarcinoma. Bisphenol A (BPA is an estrogenic chemical that is highly employed in the manufacture of a wide range of consumer products. Some observational studies have suggested that the amounts of BPA to which we are exposed could alter the reproductive organs of developing rodents. We examined the influence of BPA at low doses to address the questions of (a whether in utero exposure affects the vagina of the offspring and (b which mechanisms cause the toxic effects. Gravid Sprague-Dawley dams were administered either 0.1 (low dose or 50 mg/kg per day BPA, the no observed effect level, or 0.2 mg/kg per day 17αethinyl estradiol by gavage. Striking morphological changes were observed in the vagina of postpubertal offspring leading us to examine vaginal estrogen receptor (ER expression because BPA binds to the ERα, which is important for growth of the vaginal epithelium. We show that the full-length ERα is not expressed during estrus in the vagina of female offspring exposed to either dose of BPA when compared to the control group, whereas ERα expression does not differ from the control group during the diestrus stage. ERa downregulation seems to be responsible for the observed altered vaginal morphology.

  17. [Determination of estrogen residues in drinking water by on-line solid phase extraction based on sol-gel technique coupled with high performance liquid chromatography].

    Li, Longfei; Su, Min; Shi, Xiaolei; Wang, Yana; Wang, Minmin; He, Jinxing


    A method for the determination of diethylstilbestrol (DES), hexestrol (HEX) and dienestrol (DS) residues in drinking water was established by on-line solid phase extraction (SPE) coupled with high performance liquid chromatography (HPLC). The material synthesized on the base of sol-gel technology was employed as adsorbent. This material was prepared using 3-aminopropyltriethoxysilane (APTES) as the functional monomer, tetraethoxysilane (TEOS) as the crosslinking agent, and acetic acid as the initiator. The synthesized adsorbent showed outstanding property for the estrogen extraction. The estrogen can be caught effectively from water samples and the extraction can be achieved rapidly. Some important parameters, such as pH of sample solution, eluent solvents, loading flow rate, which might influence extraction efficiency, were optimized. The results indicated that the limit of detection (S/N = 3) of the developed method could reach 0.07-0.13 microg/L under the conditions of pH 7.0 of sample solution, methanol and 1% (v/v) acetic acid aqueous solution as the eluent solvent and the loading flow rate of 2 mL/min. The recoveries of the three estrogens from the water samples at three spiked levels ranged from 82.31% to 99.43% with RSD of 1.61%-7.15%. The method was simple, rapid, and suitable to detect the trace residues of estrogens in drinking water.

  18. [Endocrine disruptors: echoes of congress of Endocrinology in 2012].

    Nassouri, A S; Archambeaud, F; Desailloud, R


    The increased prevalence of certain diseases, along with the development of new technologies and industrialization raised the possibility of the involvement of environmental factors, industrial products, nutritional factors, infections, drugs... and endocrine disruptors. These factors may interfere via signaling pathways specific to the organism. Endocrine Disrupting Chemicals (EDCs) have been redefined by the Endocrine Society in 2012 as "exogenous chemical, or mixture of chemicals, that can interfere with any aspect of hormone action". They have therefore potentially deleterious effects on development, growth, metabolism, reproduction, the nervous, immune and cardiovascular systems. Therefore, they constitute a real public health issue. Their long half-life may explain delayed effects and their often lipophilic character may promote maternofetal transmission. Except diethylstilbestrol (DES), few formal proofs have been made on the direct role of EDCs ; arguments are based on cross-sectional studies, in vitro models and animal models. Basic research puts insight into mechanisms of action of EDCs but many questions remain unanswered. Epidemiological data are difficult to interpret because of interindividual differences in susceptibility to EDCs and of nonlinear/nonmonotonique action (as opposed to toxic dose effect), multiple interactions between environmental agents (additive effects and/or synergistic and/or antagonists), the role of the window of exposure, latency, and the possibility of transgenerational effects.

  19. On the rumors about the silent spring: review of the scientific evidence linking occupational and environmental pesticide exposure to endocrine disruption health effects

    Cocco Pierluigi


    Full Text Available Occupational exposure to some pesticides, and particularly DBCP and chlordecone, may adversely affect male fertility. However, apart from the therapeutic use of diethylstilbestrol, the threat to human reproduction posed by "endocrine disrupting" environmental contaminants has not been supported by epidemiological evidence thus far. As it concerns other endocrine effects described in experimental animals, only thyroid inhibition following occupational exposure to amitrole and mancozeb has been confirmed in humans. Cancer of the breast, endometrium, ovary, prostate, testis, and thyroid are hormone-dependent, which fostered research on the potential risk associated with occupational and environmental exposure to the so-called endocrine-disrupting pesticides. The most recent studies have ruled out the hypothesis of DDT derivatives as responsible for excess risks of cancer of the reproductive organs. Still, we cannot exclude a role for high level exposure to o,p'-DDE, particularly in post-menopausal ER+ breast cancer. On the other hand, other organochlorine pesticides and triazine herbicides require further investigation for a possible etiologic role in some hormone-dependent cancers.

  20. Endocrine disruptors and female cancer: Informing the patients (Review).

    Del Pup, Lino; Mantovani, Alberto; Luce, Amalia; Cavaliere, Carla; Facchini, Gaetano; Di Francia, Raffaele; Caraglia, Michele; Berretta, Massimiliano


    Pollutants altering the endocrine system, known as endocrine disruptors (ED), may modify the risk of female cancers. The carcinogenic effect of ED on humans has been confirmed by experimental studies for various substances including pesticides, DDT, dioxins, phthalates, bisphenol A, diethylstilbestrol, as well as heavy metals, but it is difficult to quantify precisely for several reasons hereby reviewed. Carcinogenesis is a complex and multifactorial mechanism that manifests itself over a long period of time, making difficult the detection of the specific contribution of the pollutants, whose absorbed dose is often unknown. The combined effect of various substances leads to complex interactions whose outcome is difficult to predict. These substances may accumulate and carry out their harmful effect on critical periods of life, probably also at doses considered harmless to an adult. ED can also have epigenetic adverse effects on the health of future generations. In conclusion, the carcinogenic effects of endocrine disruptors on female cancer types is plausible although additional studies are needed to clarify their mechanisms and entities. In the last part of the review we suggest ways to reduce ED exposure as it is mandatory to implement necessary measures to limit exposure, particularly during those periods of life most vulnerable to the impact of oncogenic environmental causes, such as the embryonic period and puberty.

  1. Dispersive liquid-liquid microextraction for four phenolic environmental estrogens in water samples followed by determination using capillary electrophoresis.

    Liu, Junying; Lu, Wenhui; Liu, Huitao; Wu, Xiaqing; Li, Jinhua; Chen, Lingxin


    Dispersive liquid-liquid microextraction (DLLME) coupled with CE was successfully developed for simultaneous determination of four types of phenolic environmental estrogens (PEEs), namely hexestrol (HS), bisphenol A (BPA), diethylstilbestrol (DES) and dienestrol (DS). Several parameters affecting DLLME and CE conditions were systematically investigated including the type and volume of extraction solvent and dispersive solvent, extraction time, salt, pH value, surfactant, buffer solution and so on. Under the optimal conditions, DLLME-CE exhibited strong enrichment ability, presenting high enrichment factors of 467, 241, 367 and 362 for HS, BPA, DES and DS, respectively, as well as low detection limits of 0.3, 0.6, 0.6 and 0.3 μg/L, respectively. Excellent linearity was achieved in the range of 2.0-150 μg/L for HS and DS, and 4.0-300 μg/L for BPA and DES, with correlation coefficients R>0.9983. Recoveries ranging from 70.4 to 108.1% were obtained with tap water, lake water and seawater samples spiked at three concentration levels and the relative standard deviations (RSDs, for n = 5) were 2.1-9.7%. This DLLME-CE method with high selectivity and sensitivity, high stability, simplicity, cost-effectiveness, eco-friendliness was proved potentially applicable for the rapid and simultaneous determination of PEEs in complicated water samples.

  2. Obstetrical complications and subsequent schizophrenia in adolescent and young adult offsprings: is there a relationship?

    Boog, Georges


    Schizophrenia is a psychiatric disease affecting around 1% of the population, the negative signs of which are correlated with inactivity of the prefrontal dorsolateral cortex, while an increased, more deeply localized, activity in the mesolimbic pathway may explain the positive signs. Several events occurring during pregnancy are likely to be involved in its genesis: hormonal supplementation by diethylstilbestrol, severe maternal denutrition, exposure to influenza virus, repeated psychological stress. From multicentric studies and meta-analyses in the psychiatric literature, the risk of schizophrenia appears to be multiplied by two if pregnancy is complicated, mainly by diabetes, Rhesus incompatibility, bleeding, preeclampsia, premature rupture of membranes and preterm birth. When delivery is linked to an abnormal presentation or happens via a caesarean birth for acute foetal distress, the time when the first signs of psychosis appear seems to be earlier in adolescence or in early adulthood. Cerebral imaging of schizophrenic patients shows ventriculomegaly and gray matter reduction, mainly in hippocampal volumes and in the dorsolateral prefrontal cortex. Similar alterations in the neuronal pathways have been experimentally reproduced in rats after repeated prenatal stress and perinatal hypoxia. A region on the distal portion of chromosome 1 has shown evidence for linkage to schizophrenia. Therefore, a two factor model seems to be able to explain the onset of schizophrenia in which obstetrical complications may interact with a genetic liability and in which the consequences of hypoxic events may lie on a continuum ranging from cerebral palsy in some children to subtle cognitive and behavioural disturbances in others.

  3. Risk Factors for Breast Cancer, Including Occupational Exposures

    Elisabete Weiderpass


    Full Text Available The knowledge on the etiology of breast cancer has advanced substantially in recent years, and several etiological factors are now firmly established. However, very few new discoveries have been made in relation to occupational risk factors. The International Agency for Research on Cancer has evaluated over 900 different exposures or agents to-date to determine whether they are carcinogenic to humans. These evaluations are published as a series of Monographs ( For breast cancer the following substances have been classified as “carcinogenic to humans” (Group 1: alcoholic beverages, exposure to diethylstilbestrol, estrogen-progestogen contraceptives, estrogen-progestogen hormone replacement therapy and exposure to X-radiation and gamma-radiation (in special populations such as atomic bomb survivors, medical patients, and in-utero exposure. Ethylene oxide is also classified as a Group 1 carcinogen, although the evidence for carcinogenicity in epidemiologic studies, and specifically for the human breast, is limited. The classification “probably carcinogenic to humans” (Group 2A includes estrogen hormone replacement therapy, tobacco smoking, and shift work involving circadian disruption, including work as a flight attendant. If the association between shift work and breast cancer, the most common female cancer, is confirmed, shift work could become the leading cause of occupational cancer in women.

  4. Estrogenic Activities of Fatty Acids and a Sterol Isolated from Royal Jelly

    Isohama, Yoichiro; Maruyama, Hiroe; Yamada, Yayoi; Narita, Yukio; Ohta, Shozo; Araki, Yoko; Miyata, Takeshi; Mishima, Satoshi


    We have previously reported that royal jelly (RJ) from honeybees (Apis mellifera) has weak estrogenic activity mediated by interaction with estrogen receptors that leads to changes in gene expression and cell proliferation. In this study, we isolated four compounds from RJ that exhibit estrogenic activity as evaluated by a ligand-binding assay for the estrogen receptor (ER) β. These compounds were identified as 10-hydroxy-trans-2-decenoic acid, 10-hydroxydecanoic acid, trans-2-decenoic acid and 24-methylenecholesterol. All these compounds inhibited binding of 17β-estradiol to ERβ, although more weakly than diethylstilbestrol or phytoestrogens. However, these compounds had little or no effect on the binding of 17β-estradiol to ERα. Expression assays suggested that these compounds activated ER, as evidenced by enhanced transcription of a reporter gene containing an estrogen-responsive element. Treatment of MCF-7 cells with these compounds enhanced their proliferation, but concomitant treatment with tamoxifen blocked this effect. Exposure of immature rats to these compounds by subcutaneous injection induced mild hypertrophy of the luminal epithelium of the uterus, but was not associated with an increase in uterine weight. These findings provide evidence that these compounds contribute to the estrogenic effect of RJ. PMID:18830443

  5. Estrogenic Activities of Fatty Acids and a Sterol Isolated from Royal Jelly

    Kazu-Michi Suzuki


    Full Text Available We have previously reported that royal jelly (RJ from honeybees (Apis mellifera has weak estrogenic activity mediated by interaction with estrogen receptors that leads to changes in gene expression and cell proliferation. In this study, we isolated four compounds from RJ that exhibit estrogenic activity as evaluated by a ligand-binding assay for the estrogen receptor (ER β. These compounds were identified as 10-hydroxy-trans-2-decenoic acid, 10-hydroxydecanoic acid, trans-2-decenoic acid and 24-methylenecholesterol. All these compounds inhibited binding of 17β-estradiol to ERβ, although more weakly than diethylstilbestrol or phytoestrogens. However, these compounds had little or no effect on the binding of 17β-estradiol to ERα. Expression assays suggested that these compounds activated ER, as evidenced by enhanced transcription of a reporter gene containing an estrogen-responsive element. Treatment of MCF-7 cells with these compounds enhanced their proliferation, but concomitant treatment with tamoxifen blocked this effect. Exposure of immature rats to these compounds by subcutaneous injection induced mild hypertrophy of the luminal epithelium of the uterus, but was not associated with an increase in uterine weight. These findings provide evidence that these compounds contribute to the estrogenic effect of RJ.

  6. Assessing estrogenic activity in surface water and sediment of the Liao River system in northeast China using combined chemical and biological tools

    Wang Li [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, Guangzhou 510640 (China); Ying Guangguo, E-mail: [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, Guangzhou 510640 (China); Zhao Jianliang; Liu Shan; Yang Bin; Zhou Lijun; Tao Ran; Su Haochang [State Key Laboratory of Organic Geochemistry, Guangzhou Institute of Geochemistry, Chinese Academy of Sciences, Guangzhou 510640 (China)


    This paper investigated some selected estrogenic compounds (4-t-octylphenol: 4-t-OP; 4-nonylphenols: 4-NP; bisphenol-A: BPA; diethylstilbestrol: DES; estrone: E1; 17{beta}-estradiol: E2; 17{alpha}-Ethinylestradiol: EE2; triclosan: TCS) and estrogenicity in the Liao River system using the combined chemical and in vitro yeast screen bioassay and assessed their ecological risks to aquatic organisms. The estrogenic compounds 4-t-OP, 4-NP, BPA, E1, E2 and TCS were detected in most of the samples, with their concentrations up to 52.1 2065.7, 755.6, 55.8, 7.4 and 81.3 ng/L in water, and up to 8.6, 558.4, 33.8, 7.9,

  7. A Rat α-Fetoprotein Binding Activity Prediction Model to Facilitate Assessment of the Endocrine Disruption Potential of Environmental Chemicals.

    Hong, Huixiao; Shen, Jie; Ng, Hui Wen; Sakkiah, Sugunadevi; Ye, Hao; Ge, Weigong; Gong, Ping; Xiao, Wenming; Tong, Weida


    Endocrine disruptors such as polychlorinated biphenyls (PCBs), diethylstilbestrol (DES) and dichlorodiphenyltrichloroethane (DDT) are agents that interfere with the endocrine system and cause adverse health effects. Huge public health concern about endocrine disruptors has arisen. One of the mechanisms of endocrine disruption is through binding of endocrine disruptors with the hormone receptors in the target cells. Entrance of endocrine disruptors into target cells is the precondition of endocrine disruption. The binding capability of a chemical with proteins in the blood affects its entrance into the target cells and, thus, is very informative for the assessment of potential endocrine disruption of chemicals. α-fetoprotein is one of the major serum proteins that binds to a variety of chemicals such as estrogens. To better facilitate assessment of endocrine disruption of environmental chemicals, we developed a model for α-fetoprotein binding activity prediction using the novel pattern recognition method (Decision Forest) and the molecular descriptors calculated from two-dimensional structures by Mold² software. The predictive capability of the model has been evaluated through internal validation using 125 training chemicals (average balanced accuracy of 69%) and external validations using 22 chemicals (balanced accuracy of 71%). Prediction confidence analysis revealed the model performed much better at high prediction confidence. Our results indicate that the model is useful (when predictions are in high confidence) in endocrine disruption risk assessment of environmental chemicals though improvement by increasing number of training chemicals is needed.

  8. A Rat α-Fetoprotein Binding Activity Prediction Model to Facilitate Assessment of the Endocrine Disruption Potential of Environmental Chemicals

    Huixiao Hong


    Full Text Available Endocrine disruptors such as polychlorinated biphenyls (PCBs, diethylstilbestrol (DES and dichlorodiphenyltrichloroethane (DDT are agents that interfere with the endocrine system and cause adverse health effects. Huge public health concern about endocrine disruptors has arisen. One of the mechanisms of endocrine disruption is through binding of endocrine disruptors with the hormone receptors in the target cells. Entrance of endocrine disruptors into target cells is the precondition of endocrine disruption. The binding capability of a chemical with proteins in the blood affects its entrance into the target cells and, thus, is very informative for the assessment of potential endocrine disruption of chemicals. α-fetoprotein is one of the major serum proteins that binds to a variety of chemicals such as estrogens. To better facilitate assessment of endocrine disruption of environmental chemicals, we developed a model for α-fetoprotein binding activity prediction using the novel pattern recognition method (Decision Forest and the molecular descriptors calculated from two-dimensional structures by Mold2 software. The predictive capability of the model has been evaluated through internal validation using 125 training chemicals (average balanced accuracy of 69% and external validations using 22 chemicals (balanced accuracy of 71%. Prediction confidence analysis revealed the model performed much better at high prediction confidence. Our results indicate that the model is useful (when predictions are in high confidence in endocrine disruption risk assessment of environmental chemicals though improvement by increasing number of training chemicals is needed.

  9. Identification of Physiologically Active Substances as Novel Ligands for MRGPRD

    Makiko Uno


    Full Text Available Mas-related G-protein coupled receptor member D (MRGPRD is a G protein-coupled receptor (GPCR which belongs to the Mas-related GPCRs expressed in the dorsal root ganglia (DRG. In this study, we investigated two novel ligands in addition to beta-alanine: (1 beta-aminoisobutyric acid, a physiologically active substance, with which possible relation to tumors has been seen together with beta-alanine; (2 diethylstilbestrol, a synthetic estrogen hormone. In addition to the novel ligands, we found that transfection of MRGPRD leads fibroblast cells to form spheroids, which would be related to oncogenicity. To understand the MRGPRD novel character, oncogenicity, a large chemical library was screened in order to obtain MRGPRD antagonists to utilize in exploring the character. The antagonist in turn inhibited the spheroid proliferation that is dependent on MRGPRD signaling as well as MRGPRD signals activated by beta-alanine. The antagonist, a small-molecule compound we found in this study, is a potential anticancer agent.

  10. Risk factors for breast cancer, including occupational exposures.

    Weiderpass, Elisabete; Meo, Margrethe; Vainio, Harri


    The knowledge on the etiology of breast cancer has advanced substantially in recent years, and several etiological factors are now firmly established. However, very few new discoveries have been made in relation to occupational risk factors. The International Agency for Research on Cancer has evaluated over 900 different exposures or agents to-date to determine whether they are carcinogenic to humans. These evaluations are published as a series of Monographs ( For breast cancer the following substances have been classified as "carcinogenic to humans" (Group 1): alcoholic beverages, exposure to diethylstilbestrol, estrogen-progestogen contraceptives, estrogen-progestogen hormone replacement therapy and exposure to X-radiation and gamma-radiation (in special populations such as atomic bomb survivors, medical patients, and in-utero exposure). Ethylene oxide is also classified as a Group 1 carcinogen, although the evidence for carcinogenicity in epidemiologic studies, and specifically for the human breast, is limited. The classification "probably carcinogenic to humans" (Group 2A) includes estrogen hormone replacement therapy, tobacco smoking, and shift work involving circadian disruption, including work as a flight attendant. If the association between shift work and breast cancer, the most common female cancer, is confirmed, shift work could become the leading cause of occupational cancer in women.

  11. Constant expression of cyclooxygenase-2 gene in prostate and the lower urinary tract of estrogen-treated male rats.

    Luo, C; Strauss, L; Ristimäki, A; Streng, T; Santti, R


    Expression of cyclooxygenase-2 (E. C. in prostate and the lower urinary tract (LUT) of the neonatally estrogenized male rat has been studied by using a COX-2's PCR fragment of 724 nt spanning 3 introns and a 478nt internal standard for quantitative RT-PCR. The same fragment of 724 nt was used for RNA probe in Northern hybridization. Neonatal estrogenization (10 microg/day of diethylstilbestrol on days 1-5) had no effect on COX-2 expression in prostatic urethra, prostatic lobes, or bladder. Acute estrogen treatment of castrated animals did not induce COX-2 expression, either. In addition the differential expression of basal level of COX-2 in the different lobes of prostate in normal rat was demonstrated. Our results suggest a constant expression of COX-2 gene in prostate and the lower urinary tract of the neonatally estrogenized (neoDES) rats. The present study indicates that the increased expression of COX-2 is probably not essential for the estrogen-driven development of stromal inflammation or hyperplastic and dysplastic alterations in the prostate of neoDES rats.

  12. Endocrine disruption of oestrogen action and female reproductive tract cancers.

    Gibson, Douglas A; Saunders, Philippa T K


    Endocrine disrupting chemicals (EDC) are ubiquitous and persistent compounds that have the capacity to interfere with normal endocrine homoeostasis. The female reproductive tract is exquisitely sensitive to the action of sex steroids, and oestrogens play a key role in normal reproductive function. Malignancies of the female reproductive tract are the fourth most common cancer in women, with endometrial cancer accounting for most cases. Established risk factors for development of endometrial cancer include high BMI and exposure to oestrogens or synthetic compounds such as tamoxifen. Studies on cell and animal models have provided evidence that many EDC can bind oestrogen receptors and highlighted early life exposure as a window of risk for adverse lifelong effects on the reproductive system. The most robust evidence for a link between early life exposure to EDC and adverse reproductive health has come from studies on women who were exposed in utero to diethylstilbestrol. Demonstration that EDC can alter expression of members of the HOX gene cluster highlights one pathway that might be vulnerable to their actions. In summary, evidence for a direct link between EDC exposure and cancers of the reproductive system is currently incomplete. It will be challenging to attribute causality to any single EDC when exposure and development of malignancy may be separated by many years and influenced by lifestyle factors such as diet (a source of phytoestrogens) and adiposity. This review considers some of the evidence collected to date.

  13. Effects of endocrine modulators on sexual differentiation and reproductive function in male Japanese quail.

    Halldin, Krister; Axelsson, Jeanette; Brunström, Björn


    A number of environmental contaminants have been shown to interfere with the endocrine system. Many of these compounds bind to estrogen receptors, thereby potentially disrupting estrogen-regulated functions. In this paper, we review some background data on avian sexual differentiation and present some of the results from our studies on effects of estrogenic chemicals administered during sexual differentiation in the Japanese quail. Initially, our goal was to elucidate whether a decreased male sexual behavior in quail is a suitable endpoint for studying long-term effects of exposure to estrogenic compounds during sexual differentiation in ovo. We thereafter tested some environmental pollutants, suspected to act via mimicking estrogens, using the test system developed. Results from our studies on the synthetic estrogens ethinylestradiol and diethylstilbestrol, as well as the environmental pollutants bisphenol A, tetrabromobisphenol A, and o,p'-DDT are reviewed in this paper. We conclude that the Japanese quail is well suited as an animal model for studying various long-term effects after embryonic exposure to estrogenic compounds. Depressed sexual behavior proved to be the most sensitive of the variables studied in males and we find this endpoint appropriate for studying effects of endocrine modulating chemicals in the adult quail following embryonic exposure.

  14. Effects of dietary phytoestrogens in vivo and in vitro in rainbow trout and Siberian sturgeon: interests and limits of the in vitro studies of interspecies differences.

    Latonnelle, K; Le Menn, F; Kaushik, S J; Bennetau-Pelissero, C


    A study of the effects of dietary genistein on trout and sturgeon in vivo showed that sturgeon was sensitive to 20 ppm of genistein, whereas trout was not. To analyze the origin of this interspecies difference in sensitivity, a cell culture technique was developed with hepatocytes from sturgeon and compared to results obtained with hepatocytes from trout in the same system. The hepatocyte culture proved to be useful as bioassay for estrogenicity. Vitellogenin (VTG), assayed by a specific enzyme-linked immunosorbent assay, was used as a biomarker of the estrogenic activity. 17 beta-Estradiol, its glucuronide and sulfate derivatives, and estradiol analogues (ethynylestradiol and diethylstilbestrol) were tested. Nonestrogenic compounds such as androgens, progesterone, and cortisol were tested as negative controls. VTG production was monitored at doses ranging from 1 nM to 10 microM estradiol. Phytoestrogens, from the isoflavone family, were tested individually at increasing doses exhibiting dose response curves for concentrations from 500 nM to 10 microM. With tamoxifen, an antagonist of estrogen receptors, the estrogenic effect was partially reduced. The effect was the same with ICI182,780 in sturgeon, whereas the effect was the opposite in trout. The estrogenic potency of the isoflavones ranged differently between the two species in the following order: biochanin A vitro, whereas its activity was weakest in vivo. These data suggest that one must reconsider the relevance of heterologous estrogenic tests and of homologous in vitro tests for estrogenic potency of chemicals.

  15. Occurrence and removal of phenolic endocrine disrupting chemicals in the water treatment processes

    Lv, Xuemin; Xiao, Sanhua; Zhang, Gang; Jiang, Pu; Tang, Fei


    This paper evaluated the occurrence and removal efficiency of four selected phenolic endocrine disrupting chemicals (bisphenol A (BPA), octylphenol (OP), nonylphenol (NP) and diethylstilbestrol (DES)) in two drinking waterworks in Jiangsu province which take source water from Taihu Lake. The recombined yeast estrogen screen (YES) and liquid chromatography tandem mass spectrometry (LC-MS/MS) were applied to assess the estrogenicity and detect the estrogens in the samples. The estrogen equivalents (EEQs) ranged from nd (not detected) to 2.96 ng/L, and the estrogenic activities decreased along the processes. Among the 32 samples, DES prevailed in all samples, with concentrations ranging 1.46-12.0 ng/L, BPA, OP and NP were partially detected, with concentrations ranging from nd to 17.73 ng/L, nd to 0.49 ng/L and nd to 3.27 ng/L, respectively. DES was found to be the main contributor to the estrogenicity (99.06%), followed by NP (0.62%), OP (0.23%) and BPA (0.09%). From the observation of treatment efficiency, the advanced treatment processes presented much higher removal ratio in reducing DES, the biodegradation played an important role in removing BPA, ozonation and pre-oxidation showed an effective removal on all the four estrogens; while the conventional ones can also reduce all the four estrogens.

  16. Early programing of uterine tissue by bisphenol A: Critical evaluation of evidence from animal exposure studies.

    Suvorov, Alexander; Waxman, David J


    Exposure to Bisphenol A (BPA) during the critical window of uterine development has been proposed to program the uterus for increased disease susceptibility based on well-documented effects of the potent xenoestrogen diethylstilbestrol. To investigate this proposal, we reviewed 37 studies of prenatal and/or perinatal BPA exposure in animal models and evaluated evidence for: molecular signatures of early BPA exposure; the development of adverse uterine health effects; and epigenetic changes linked to long-term dysregulation of uterine gene expression and health effects. We found substantial evidence for adult uterine effects of early BPA exposure. In contrast, experimental support for epigenetic actions of early BPA exposure is very limited, and largely consists of effects on Hoxa gene DNA methylation. Critical knowledge gaps were identified, including the need to fully characterize short-term and long-term uterine gene responses, interactions with estrogens and other endogenous hormones, and any long-lasting epigenetic signatures that impact adult disease. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Comparative study of bisphenol A and its analogue bisphenol S on human hepatic cells: a focus on their potential involvement in nonalcoholic fatty liver disease.

    Peyre, Ludovic; Rouimi, Patrick; de Sousa, Georges; Héliès-Toussaint, Cécile; Carré, Benjamin; Barcellini, Sylvie; Chagnon, Marie-Christine; Rahmani, Roger


    For several decades, people have been in contact with bisphenol A (BPA) primarily through their diet. Nowadays it is gradually replaced by an analogue, bisphenol S (BPS). In this study, we compared the effects of these two bisphenols in parallel with the positive control diethylstilbestrol (DES) on different hepatocyte cell lines. Using a cellular impedance system we have shown that BPS is less cytotoxic than BPA in acute and chronic conditions. We have also demonstrated that, contrary to BPA, BPS is not able to induce an increase in intracellular lipid and does not activate the PXR receptor which is known to be involved in part, in this process. In parallel, it failed to modulate the expression of CYP3A4 and CYP2B6, the drug transporter ABCB1 and other lipid metabolism genes (FASN, PLIN). However, it appears to have a weak effect on GSTA4 protein expression and on the Erk1/2 pathway. In conclusion, in contrast to BPA, BPS does not appear to induce the metabolic syndrome that may lead to non-alcoholic fatty liver disease (NAFLD), in vitro. Although we have to pay special attention to BPS, its use could be less dangerous concerning this toxicological endpoint for human health.

  18. Pregnancy-dependent initiation in tumorigenesis of Wistar rat mammary glands by sup 60 Co-irradiation

    Inano, Hiroshi; Suzuki, Keiko; Ishii-Ohba, Hiroko; Ikeda, Kiyomi (National Inst. of Radiological Sciences, Chiba (Japan)); Wakabayashi, Katsumi (Gunma Univ., Maebashi (Japan). Hormone Assay Center)


    Pregnant Wistar rats received whole body irradiation with 260 cGy {gamma}-rays at days 7, 14 and 20 of pregnancy and then were treated with diethylstilbestrol (DES) for 1 year. The highest incidence (92.9%) for tumorigenesis of mammary glands was observed in the rats irradiated in late pregnancy. Histological examination showed that tumors were classified as fibroadenoma and adenocarcinoma. To determine the reasons for specific induction of mammary tumors by irradiation in late pregnancy, hormone concentrations in serum and estrogen receptors in mammary glands during pregnancy were measured. Concentrations of estradiol, progesterone, 11-deoxycorticosterone and placental lactogen at day 20 were higher than at days 7 and/or 14, but no difference was observed in the concentrations of prolactin and thyroid-stimulating hormone during pregnancy. The estrogen receptor in mammary glands at day 20 was indicated to have the highest affinity and the highest binding capacity during pregnancy. Normal mammary glands at day 20 were suggested to have more abundant epithelial cells in the mammary lobes than those at days 7 and 14. The data suggest that the critical requirements for the initiation of tumorigenesis by {gamma}-rays are dependent upon the differentiated state of mammary glands exposed to various hormones, and that the concentration and persistence of the synthetic estrogen (DES) are necessary for the promotion of tumorigenesis of the irradiated mammary glands. (Author).

  19. Use of sub-micron sized resin particles for removal of endocrine disrupting compounds and pharmaceuticals from water and wastewater.

    Murray, Audrey; Örmeci, Banu; Lai, Edward P C


    Endocrine disrupting compounds (EDCs) and pharmaceuticals pose a challenge for water and wastewater treatment because they exist at very low concentrations in the presence of substances at much higher concentrations competing for adsorption sites. Sub-micron sized resin particles (approximately 300nm in diameter) (SMR) were tested to evaluate their potential as a treatment for EDCs including: 17-β estradiol (E2), 17-α ethinylestradiol (EE2), estrone (E1), bisphenol A (BPA), and diethylstilbestrol (DES) as well as 12 pharmaceuticals. SMR were able to remove 98% of spiked E2, 80% of EE2, 87% of BPA, and up to 97% of DES from water. For a 0.5ppm mixture of E2, EE2, E1, BPA and DES, the minimum removal was 24% (E2) and the maximum was 49% (DES). They were also able to remove the pharmaceuticals from deionized water and wastewater. Overall, SMR are a promising advanced treatment for removal of both EDCs and pharmaceuticals. Copyright © 2016. Published by Elsevier B.V.

  20. Different patterns of developmental toxicity in the rat following prenatal administration of structurally diverse chemicals

    Simmons, D.L.; Valentine, D.M.; Bradshaw, W.S.


    Differences in the profiles of developmental toxicity for four structurally diverse chemical compounds have been defined following prenatal exposure in the rat. Diethylstilbestrol (DES), 3,4,3',4'-tetrachlorobiphenyl (4CB), zeranol, and cadmium were administered by gavage to Sprague-Dawley rats daily from d 6 through d 18 of gestation. Dams were sacrificed at four prenatal endpoints and the numbers of live and dead fetuses and resorbed embryos were counted. Additional dams were allowed to bring their litters to term, and their offspring were monitored until they reached adulthood. DES induced prenatal death primarily in early embryonic life, and also during parturition. 4CB increased mortality from late in gestation up to 24 h after birth, and altered the sex ratio of survivors by selectively acting against males in utero. Exposure to zeranol resulted in embryolethality only. Cadmium was not lethal to the conceptus at any dose below the dose that caused maternal mortality. Only 4CB had an obvious teratogenic effect, causing intestinal hemorrhage. All compounds produced transient perinatal decreases in the weight of the offspring. 30 references, 6 tables.

  1. Potentiation of the reductase activity of protein disulphide isomerase (PDI) by 19-nortestosterone, bacitracin, fluoxetine, and ammonium sulphate.

    Hassan, Maya Haj; Alvarez, Eva; Cahoreau, Claire; Klett, Danièle; Lecompte, François; Combarnous, Yves


    Protein disulphide isomerase (PDI) in the endoplasmic reticulum catalyzes the rearrangement of disulphide bridges during folding of secreted proteins. It binds various molecules that inhibit its activity. But here, we looked for molecules that would potentiate its activity. PDI reductase activity was measured in vitro using di-eosin-oxidized glutathione as substrate. Its classical inhibitor bacitracin was found to exert a biphasic effect: stimulatory at low concentrations (∼10(-6) M) and inhibitory only at higher concentrations (∼10(-4)-10(-3) M). The weak oestrogenic molecule bisphenol A was found to exert a weak inhibitory effect on PDI reductase activity relative to the strong oestrogens, ethynylestradiol, and diethylstilbestrol. Like 19-nortestosterone, fluoxetine was found to exert a potentiating effect on PDI reductase activity and their potentiating effects could be reversed by increasing concentrations of oestrogens. In conclusion, this paper provides the first identification of potentiators of PDI activity that are potential pharmaceuticals against pathologies affecting protein folding such as Alzheimer's disease.

  2. Introduction and overview. Perinatal carcinogenesis: growing a node for epidemiology, risk management, and animal studies.

    Anderson, Lucy M


    Perinatal carcinogenesis as a cross-disciplinary concern is the subject of this special issue of Toxicology and Applied Pharmacology, which consists of a total of eight reviews or commentaries in the areas of epidemiology, risk assessment, and animal models. Some of the conclusions from these articles, and the Questions and Answers section that follows most of them, are summarized here. There is adequate reason to suspect that perinatal exposures contribute to human cancer risk, both childhood cancers, and those appearing later in life. The latter type of risk may actually be quantitatively the more important, and involve a wide range of types of effects, but has received only limited attention. With regard to childhood cancers, fetal irradiation and diethylstilbestrol exposure are known etiological agents, and it is likely, but not yet certain, there are additional external causes of a portion of these. Some current focal points of interest here include nitroso compounds, DNA topoisomerase inhibitors, viruses, anti-AIDS drugs, and endocrine disruptors. Regulatory agencies must rely heavily on animal data for estimation of human risk due to perinatal exposures to chemicals, and the quantity and quality of these data presently available for this purpose are greatly limiting. Correctly designed conventional animal studies with suspect chemicals are still needed. Furthermore, genetically engineered mouse models for childhood cancers, especially medulloblastoma, have become available, and could be used for screening of candidate causative agents for this cancer type, and for better understanding of gene-environment interactions.

  3. Validation of a new yeast-based reporter assay consisting of human estrogen receptors alpha/beta and coactivator SRC-1: application for detection of estrogenic activity in environmental samples.

    Chu, Wai-Ling; Shiizaki, Kazuhiro; Kawanishi, Masanobu; Kondo, Mami; Yagi, Takashi


    Endocrine disruptors are exogenous substances that act like hormones in the endocrine system and disrupt the physiologic function of endogenous hormones. In the present study, we established reporter yeast strains (Saccharomyces cerevisiae) expressing human estrogen receptors, ERalpha or ERbeta. These strains contain a reporter plasmid carrying an estrogen responsive element (ERE) upstream of the beta-galactosidase gene, and a plasmid expressing a steroid receptor coactivator, SRC-1e. Using these reporter strains, we demonstrated dose-dependent estrogenic activities of different categories of ligands, a natural hormone, 17beta-estradiol (E2); a synthetic drug, diethylstilbestrol (DES); phytoestrogens, genistein, daizein and emodin; and an environmental endocrine disrupter, bisphenol A. EC(50) values of E2 for ERalpha and ERbeta are 5.31 x 10(-10) and 5.85 x 10(-10) M, respectively. We also demonstrated that these yeasts were applicable for measuring estrogenic activities of environmental water samples. Most downstream sites of a river showed similar activity in both ERalpha and ERbeta assays. These yeast strains are useful and convenient for detecting and comparing the estrogenic ligand activities of environmental samples in response to ERalpha and ERbeta.

  4. Steroidal hormones and other endocrine active compounds in shallow groundwater in nonagricultural areas of Minnesota—Study design, methods, and data, 2009–10

    Erickson, Melinda L.


    The U.S. Geological Survey, in cooperation with the Minnesota Pollution Control Agency, completed a study on the occurrence of steroidal hormones and other endocrine active compounds in shallow groundwater in nonagricultural areas of Minnesota during 2009–10. This report describes the study design and methods, and presents the data collected on steroidal hormones and other related compounds. Environmental and quality-control samples were collected from 40 wells as part of this study. Samples were analyzed by the U.S. Geological Survey National Water Quality Laboratory for 16 steroidal hormones and 4 other related compounds, of which all but 2 compounds are endocrine active compounds. Most of the water samples did not contain detectable concentrations of any of the 20 compounds analyzed. Water samples from three wells had detectable concentrations of one or more compounds. Bisphenol A was detected in samples from three wells, and trans-diethylstilbestrol was detected in one of the samples in which bisphenol A also was detected.

  5. Dual cloud point extraction coupled with hydrodynamic-electrokinetic two-step injection followed by micellar electrokinetic chromatography for simultaneous determination of trace phenolic estrogens in water samples.

    Wen, Yingying; Li, Jinhua; Liu, Junshen; Lu, Wenhui; Ma, Jiping; Chen, Lingxin


    A dual cloud point extraction (dCPE) off-line enrichment procedure coupled with a hydrodynamic-electrokinetic two-step injection online enrichment technique was successfully developed for simultaneous preconcentration of trace phenolic estrogens (hexestrol, dienestrol, and diethylstilbestrol) in water samples followed by micellar electrokinetic chromatography (MEKC) analysis. Several parameters affecting the extraction and online injection conditions were optimized. Under optimal dCPE-two-step injection-MEKC conditions, detection limits of 7.9-8.9 ng/mL and good linearity in the range from 0.05 to 5 μg/mL with correlation coefficients R(2) ≥ 0.9990 were achieved. Satisfactory recoveries ranging from 83 to 108% were obtained with lake and tap water spiked at 0.1 and 0.5 μg/mL, respectively, with relative standard deviations (n = 6) of 1.3-3.1%. This method was demonstrated to be convenient, rapid, cost-effective, and environmentally benign, and could be used as an alternative to existing methods for analyzing trace residues of phenolic estrogens in water samples.

  6. Effects of pentosan polysulfate sodium on the estrogen-induced pituitary prolactinoma in Fischer 344 rats.

    Mucha, Slawomir; Melen-Mucha, Gabriela; Stepien, Tomasz; Godlewski, Andrzej; Stepien, Henryk


    The development of estrogen-induced pituitary prolactinoma in Fischer 344 (F344) rats is associated with enhanced neovascularization. This type of tumor is a rich source of basic fibroblast growth factor (bFGF), which possesses strong mitogenic and angiogenic properties. Pentosan polysulfate sodium (PPS) has been shown to exert antitumor activity by antagonizing the binding of bFGF to cell surface receptors. We have examined the effects of pentosan on tumor growth, hyperprolactinemia and angiogenesis in diethylstilbestrol-induced anterior pituitary adenoma in F344 rats. Chronic treatment with PPS did not cause any changes in the pituitary weight and serum prolactin concentration in comparison with untreated animals. The density of microvessels identified by CD-31 was also not affected by the tested drug. On the other hand, pentosan has been found to decrease cell proliferation evaluated by a number of PCNA-positive stained cell nuclei. Moreover, the TUNEL method has revealed an increased number of apoptotic bodies within the anterior pituitary after treatment with PPS. Despite the antiproliferative and proapoptotic activity of pentosan, the drug failed to inhibit tumor growth. This fact might be due to the lack of antiangiogenic activity of PPS in this experimental design.

  7. Thin metal organic frameworks coatings by cathodic electrodeposition for solid-phase microextraction and analysis of trace exogenous estrogens in milk.

    Lan, Hangzhen; Pan, Daodong; Sun, Yangying; Guo, Yuxing; Wu, Zhen


    Cathodic electrodeposition (CED) has received great attention in metal-organic frameworks (MOFs) synthesis due to its distinguished properties including simplicity, controllability, mild synthesis conditions, and product continuously. Here, we report the fabrication of thin (Et3NH)2Zn3(BDC)4 (E-MOF-5) film coated solid phase microextraction (SPME) fiber by a one-step in situ cathodic electrodeposition strategy. Several etched stainless steel fibers were placed in parallel in order to achieve simultaneously electrochemical polymerization. The influence of different polymerization parameters Et3NHCl concentration and polymerization time were evaluated. The proposed method requires only 20 min for the preparation of E-MOF-5 coating. The optimum coating showed excellent thermal stability and mechanical durability with a long lifetime of more than 120 repetitions SPME operations, and also exhibited higher extraction selectivity and capacity to four estrogens than commonly-used commercial PDMS coating. The limits of detection for the estrogens were 0.17-0.56 ng mL(-1). Fiber-to-fiber reproducibility (n = 8) was in the respective ranges of 3.5%-6.1% relative standard deviation (RSD) for four estrogens for triplicate measurements at 200 ng mL(-1). Finally, the (E-MOF-5) coated fiber was evaluated for ethinylestradiol (EE2), bisphenol A (BPA), diethylstilbestrol (DES), and hexestrol (HEX) extraction in the spiked milk samples. The extraction performance of this new coating was satisfied enough for repeatable use without obvious decline.

  8. Re-establishment of Spermatogenesis by Diethyistilbestrol after 2,5-Hexanedione-induced Irreversible Testicular Atrophy in Rats

    Yue QIAN; Fuqing ZENG


    To investigate the effects of diethylstilbestrol (DES) in reestablishing spermatogenesis and the mechanism by which estrogen works on spermatogenesis, rats were exposed to 1% 2,5-HD for 5 week. Then 0.1 mL of DES was given (s.c.) at a rate of 0.3 μg/kg, 30 μg/kg, 3 ms/kg every other day for 2 weeks respectively (DES group) while the other rats received ethyldeate only. Plasma testosterone (T) and LH were measured on the 8th week after the treatment. The rats were killed at the 18th week. The left testis was histopathologieally examined. In all the rats in the DES groups, spermatogenesis was re-established and the rats in the 30 μg/kg group showed the best results. Serum T was suppressed markedly in rats of 30 μg/kg and 3 mg/kg groups while T was only mildly inhibited in 0.3 μg/kg group, without significant difference found in serum LH. It is concluded that the nearly complete testicular atrophy could be reversed by DES treatment in rats. Estrogen plays an important part in spermatogenesis, and the role of estrogen in spermatogenesis is more than suppressing the hypothalamo-pituitary-testis axis.

  9. Induction and inhibition of oocyte maturation by EDCs in zebrafish

    Tokumoto Mika


    Full Text Available Abstract Background Oocyte maturation in lower vertebrates is triggered by maturation-inducing hormone (MIH, which acts on unidentified receptors on the oocyte surface and induces the activation of maturation-promoting factor (MPF in the oocyte cytoplasm. We previously described the induction of oocyte maturation in fish by an endocrine-disrupting chemical (EDC, diethylstilbestrol (DES, a nonsteroidal estrogen. Methods In this study, stimulatory and inhibitory effects of EDCs and natural steroids on oocyte maturation were examined in zebrafish. For effective agents, some details about the mechanism in induction or inhibition of maturation were examined. Possible groups of DES interacting with the MIH receptor are discussed based on relative potency of steroids to induce maturation. Results Among agents tested, tamoxifen (TAM and its metabolite 4-hydroxytamoxifen (4-OHT showed stimulatory activity similar to DES. The time courses of the change in germinal vesicle breakdown and an intracellular molecular event (the synthesis of cyclin B induced by TAM were indistinguishable from those induced by MIH. In contrast, pentachlorophenol (PCP had a potent inhibitory effect on MIH-induced oocyte maturation. PCP inhibited not only MIH-induced maturation but also DES- and TAM-induced maturation. Methoxychlor also inhibited maturation when oocytes were pre-treated with this agent. Conclusion These results suggest that EDCs act as agonists or antagonists in the induction of oocyte maturation in fish.

  10. Semiquinone formation and DNA base damage by toxic quinones and inhibition by N-acetylcysteine (NAC)

    Lewis, D.C.; Shibamoto, T.


    Toxic, mutagenic, carcinogenic, and teratogenic effects have been reported for some quinones as well as compounds metabolized to quinones. Semiquinone radical formation, thymidine degradation, and protection by NAC were studied in a hypoxanthine/xanthine oxidase (HX/XO) system. Quinone, benzo(a)pyrene-3,6-quinone, danthron, doxorubicin, emodin, juglone, menadione, and moniliformin were tested. Diethylstilbestrolquinone, N-acetylquinoneimine, and benzoquinonediimine, hypothesized toxic metabolites of diethylstilbestrol, acetaminophen and p-phenylenediamine, respectively, were synthesized and studied. Semiquinone radical formation was assessed in a HX/XO system monitoring cytochrome C reduction. Large differences in rates of semiquinone radical formation were noted for different quinones, with V/Vo values ranging from 1.2 to 10.6. DNA base degradation, thymine or thymidine glycol formation, and thiobarbituric acid reactive substance (TBARS) production were measured in a similar system containing thymine, thymidine, calf thymus DNA, or deoxyribose. TBARS formation was observed with deoxyribose, but thymidine degradation without TBARS formation was noted with thymidine. NAC (0.5 to 10 mM) caused dose-dependent inhibition of quinone-induced cytochrome C reduction.

  11. Regional differences in the prostate of the neonatally estrogenized mouse

    Pylkkaenen, L.S.; Santti, R.; Newbold, R.; McLachlan, J.A. (Univ. of Turku (Finland))


    Neonatal estrogenization of the mouse with diethylstilbestrol resulted in time-of-exposure and dose-dependent inhibition of the growth of the prostatic lobes observed at the age of 2 mon. The critical time was the days 1-6 of postnatal life. In neonatally estrogenized (neoDES) mice, responses to 5 alpha-dihydrotestosterone in terms of nuclear 3H-thymidine labelling were altered concomitantly with the inhibition of growth and were in accordance with changes in the relative volumes of epithelium, glandular lumina, and interacinar stroma. Secondary estrogen treatment of neoDES mice with 17 beta-estradiol did not increase 3H-thymidine labelling in the prostate of control or neoDES mice. However, it induced squamous epithelial metaplasia in periurethral collecting ducts and proximal parts of coagulating glands of neoDES animals. In control mice only slight epithelial hyperplasia could be observed after similar treatment. Estrogen receptors, located immunocytochemically in nuclei of stromal cell, corresponded with the sites of increased estrogen sensitivity, observed as metaplastic transformation. When the neoDES animals aged, epithelial hyperplasia and dysplasia could be observed at distinct prostatic sites, ie, the periurethral collecting ducts and the coagulating glands and periurethral glands, and stromal inflammation become more extensive. Almost identical location of the epithelial changes and the altered estrogen response is suggestive of causal relationship.

  12. In vitro study of the binding between chlorpyrfos and sex hormones using headspace solid-phase microextraction combined with high-performance liquid chromatography: A new aspect of pesticides and breast cancer risk.

    Farhadi, K; Tahmasebi, R; Biparva, P; Maleki, R


    Endocrine-disrupting chemicals are compounds that alter the normal functioning of the endocrine system. Organophosphorus insecticides, as chlorpyrifos (CPS), receive an increasing consideration as potential endocrine disrupters. Physiological estrogens, including estrone (E1), 17β-estradiol (E2), and diethylstilbestrol (DES) fluctuate with life stage, suggesting specific roles for them in biological and disease processes. There has been great interest in whether certain organophosphorus pesticides can affect the risk of breast cancer. An understanding of the interaction processes is the key to describe the fate of CPS in biological media. The objectives of this study were to evaluate total, bound, and freely dissolved amount of CPS in the presence of three estrogenic sex hormones (ESHs). In vitro experiments were conducted utilizing a headspace solid phase microextraction (HS-SPME) combined with high-performance liquid chromatography (HPLC) method. The obtained Scatchard plot based on the proposed SPME-HPLC method was employed to determine CPS-ESHs binding constant and the number of binding sites as well as binding percentage of each hormone to CPS. The number of binding sites per studied hormone molecule was 1.10, 1, and 0.81 for E1, E2, and DES, respectively. The obtained results confirmed that CPS bound to one class of binding sites on sex hormones.

  13. Antidepressant-like effect of different estrogenic compounds in the forced swimming test.

    Estrada-Camarena, Erika; Fernández-Guasti, Alonso; López-Rubalcava, Carolina


    The present study evaluated the possible antidepressant-like action of the natural estrogen 17beta-estradiol (E(2), 2.5-10 microg/rat), the synthetic steroidal estrogen ethinyl-estradiol (EE(2), 1.25-10.0 microg/rat), and the nonsteroidal synthetic estrogen, diethyl-stilbestrol (DES, 0.25-1.0 mg/rat) in ovariectomized adult female Wistar rats using the forced swimming test (FST). The behavioral profile induced by the estrogens was compared with that induced by the antidepressants fluoxetine (FLX, 2.5-10 mg/kg) and desipramine (DMI, 2.5-10 mg/kg). In addition, the temporal course of the antidepressant-like action of the estrogenic compounds was analyzed. FLX and DMI induced an antidepressant-like effect characterized by a reduced immobility and increased swimming for FLX and decreased immobility and increased climbing for DMI. Both E(2) and EE(2) produced a decrease in immobility and an increase in swimming, suggesting an antidepressant-like action. DES did not affect the responses in this animal model of depression at any dose tested. The time course analysis of the actions of E(2) (10 microg/rat) and EE(2) (5 microg/rat) showed that both compounds induced an antidepressant-like effect observed 1 h after their injection lasting for 2-3 days.

  14. The post-coital pills as over the counter drugs

    Sukhbir Singh


    Full Text Available Post-coital pill or emergency contraceptives are birth control measures that, if taken after sexual intercourse, may prevent pregnancy. High dose of postcoital contraceptives like diethylstilbestrol & other estrogens were being used for some time without any approval by FDA. Task force on postovulatory methods of fertility regulations 1998, conducted clinical trials leading to approval of two preparations for postcoital contraception by FDA namely levonorgestrel 0.75mg & combination of 0.25 levonorgestrel & 0.05 mg of ethinyl estradiol. Levonorgestrel is more popular in India because it is more effective & has fewer side effects. The first dose of levonorgestrel 0.75mg should be taken within 72 hours of unprotected intercourse followed by a second dose after 12 hours. This treatment reduces the risk of pregnancy by approximately 75%. Oral levonorgestrel acts as an emergency contraceptive principally by preventing ovulation or fertilization by altering tubal transport of sperm and/or ova. In addition, it may inhibit implantation by altering the endometrium. It is not effective once the process of implantation has begun. [Int J Basic Clin Pharmacol 2013; 2(2.000: 227-228

  15. Using Fenton Oxidation to Simultaneously Remove Different Estrogens from Cow Manure

    Minxia Sun


    Full Text Available The presence of estrogens in livestock excrement has raised concerns about their potential negative influence on animals and the overall food cycle. This is the first investigation to simultaneously remove estrogens, including estriol (E3, bisphenol A (BPA, diethylstilbestrol (DES, estradiol (E2, and ethinyl estradiol (EE2, from cow manure using a Fenton oxidation technique. Based on the residual concentrations and removal efficiency of estrogens, the Fenton oxidation reaction conditions were optimized as follows: a H2O2 dosage of 2.56 mmol/g, a Fe(II to H2O2 molar ratio of 0.125 M/M, a solid to water mass ratio of 2 g/mL, an initial pH of 3, and a reaction time of 24 h. Under these conditions, the simultaneous removal efficiencies of E3, BPA, DES, E2, and EE2, with initial concentrations in cow manure of 97.40, 96.54, 100.22, 95.01, and 72.49 mg/kg, were 84.9%, 99.5%, 99.1%, 97.8%, and 84.5%, respectively. We clarified the possible Fenton oxidation reaction mechanisms that governed the degradation of estrogens. We concluded that Fenton oxidation technique could be effective for efficient removal of estrogens in livestock excrement. Results are of great importance for cow manure reuse in agricultural management, and can be used to reduce the threat of environmental estrogens to human health and ecological safety.

  16. Human teratogens update 2011: can we ensure safety during pregnancy?

    Rasmussen, Sonja A


    Anniversaries of the identification of three human teratogens (i.e., rubella virus in 1941, thalidomide in 1961, and diethylstilbestrol in 1971) occurred in 2011. These experiences highlight the critical role that scientists with an interest in teratology play in the identification of teratogenic exposures as the basis for developing strategies for prevention of those exposures and the adverse outcomes associated with them. However, an equally important responsibility for teratologists is to evaluate whether medications and vaccines are safe for use during pregnancy so informed decisions about disease treatment and prevention during pregnancy can be made. Several recent studies have examined the safety of medications during pregnancy, including antiviral medications used to treat herpes simplex and zoster, proton pump inhibitors used to treat gastroesophageal reflux, and newer-generation antiepileptic medications used to treat seizures and other conditions. Despite the large numbers of pregnant women included in these studies and the relatively reassuring results, the question of whether these medications are teratogens remains. In addition, certain vaccines are recommended during pregnancy to prevent infections in mothers and infants, but clinical trials to test these vaccines typically exclude pregnant women; thus, evaluation of their safety depends on observational studies. For pregnant women to receive optimal care, we need to define the data needed to determine whether a medication or vaccine is "safe" for use during pregnancy. In the absence of adequate, well-controlled data, it will often be necessary to weigh the benefits of medications or vaccines with potential risks to the embryo or fetus.

  17. An azide-insensitive low-affinity ATPase stimulated by Ca2+ or Mg2+ in basal-lateral and brush border membranes of kidney cortex.

    Ilsbroux, I; Vanduffel, L; Teuchy, H; De Cuyper, M


    Basal-lateral and brush border membranes from pig kidney cortex were prepared by differential centrifugation followed by free-flow electrophoresis. In each type of membrane, azide-insensitive, low-affinity Ca2+-ATPase and Mg2+-ATPase activities are demonstrated. A comparative study for both membranes further reveals the following analogies between these ATPases: (a) they show maximal activity between pH 8 and 8.5; (b) they exhibit Km values for Ca-ATP or Mg-ATP in the millimolar range and have a comparable low substrate specificity; (c) they are insensitive to 10 microM of vanadate, N,N'-dicyclohexylcarbodiimide, e diethylstilbestrol, quercetin, harmaline and amiloride. The partial inhibition by 1 mM of the various compounds is rather aspecific. In view of these similarities it is concluded that only one enzyme entity is responsible for the activity which is measured in both membrane types. The HCO3-stimulated Mg2+-ATPase activity in pig kidney cortex was also studied. This enzyme, however, is clearly of mitochondrial origin since the HCO3-stimulation coincides with the distribution profile of succinate dehydrogenase, a mitochondrial marker; and since it is inhibited by azide.

  18. Electrochemical sensor based on magnetic molecularly imprinted nanoparticles at surfactant modified magnetic electrode for determination of bisphenol A.

    Zhu, Lili; Cao, Yuhua; Cao, Guangqun


    A selective electrochemical sensor based on magnetic molecularly imprinted nanoparticles was developed for determination of bisphenol A (BPA). The particles with regular morphology, high saturation magnetization and good monodispersion were prepared. The hydrophilicity, sensitivity and anti-fouling of the sensor were enhanced by modifying carbon paste electrode with surfactant CTAB in advanced. The results demonstrated that the response of BPA on imprinted electrode was 2.6 times as much as that on non-imprinted sensor. Moreover, the separation factors of BPA to β-estradiol, estriol and diethylstilbestrol were 16.5, 17.3 and 6.6, respectively. Under optimized conditions, the currents were found to be proportional to the BPA concentrations in the range of 6.0×10(-7)-1.0×10(-4) mol/L with a detection limit of 1.0×10(-7) mol/L (S/N=3). A rapid response of the imprinted sensor was obtained within 3 min. The developed sensor was successfully used for determination of BPA in actual samples such as drink bottles and lake water.

  19. Constitutive and heat-shock induced expression of Hsp70 mRNA during chicken testicular development and regression.

    Mezquita, B; Mezquita, J; Durfort, M; Mezquita, C


    The constitutive and heat shock induced expression of Hsp70 mRNA was investigated in normal adult chicken testis and in adult testis after testicular regression induced by diethylstilbestrol (DES) treatment. In addition to the canonical form of Hsp70 mRNA, we have detected transcripts with an extended 5'UTR and transcripts containing, in the 5'UTR, sequences of the 18S ribosomal RNA. Hsp70 was expressed in unstressed male gonads in adult and regressed testis, being the expression much lower in regressed testis. Upon heat shock at 44 degrees C or 46 degrees C, Hsp70 was highly induced in both tissues. However, when testicular seminiferous tubules were incubated at the chicken internal temperature of 39 degrees C, no induction of Hsp70 was observed in mature testis, while the expression markedly increased in regressed testis. Induction at 39 degrees C was completely inhibited in the presence of 6 mM aspirin. Aspirin in the range 3-10 mM decreases the expression of Hsp70 in unstressed and stressed testicular cells, in striking contrast with the effect observed in other tissues as liver. These data suggest that the expression of Hsp70 is regulated in a specific manner in chicken testis and particularly in the male gonad undergoing regression. Copyright 2001 Wiley-Liss, Inc.

  20. The Effect of Extracellular Components from Colletotrichum lindemuthianum on Membrane Transport in Vesicles Isolated from Bean Hypocotyl.

    Rogers, K R; Anderson, A J


    Extracellular components released from mycelia of the alpha and beta races of the bean pathogen, Colletotrichum lindemuthianum, inhibited proton uptake in sealed vesicles prepared from bean hypocotyls. Differential sensitivity of ATP-driven proton transport to nitrate, vanadate, N,N'-dicyclohexylcarbodiimide, diethylstilbestrol, and oligomycin suggested the vesicles were enriched for tonoplast. Anion stimulation of proton transport, by enhancement of ATPase activity and dissipation of the membrane potential, was consistent with this conclusion. Although fungal components inhibited the formation of a pH gradient, the membrane potential was unaffected and the ATPase activity slightly stimulated. These data suggest that the fungal components produce an electroneutral proton exchange. Proton transport in Dark Red Kidney bean tonoplast vesicles was inhibited by mycelial preparations from the incompatible alpha race and compatible beta race. Elicitor activity, however, was greater in the alpha race fractions. Elicitor purified from alpha race culture filtrate did not inhibit proton transport in vesicles isolated from Dark Red Kidney bean. Consequently, elicitor activity need not be associated with an ability to impair tonoplast function.

  1. The Effect of Extracellular Components from Colletotrichum lindemuthianum on Membrane Transport in Vesicles Isolated from Bean Hypocotyl 1

    Rogers, Kim R.; Anderson, Anne J.


    Extracellular components released from mycelia of the α and β races of the bean pathogen, Colletotrichum lindemuthianum, inhibited proton uptake in sealed vesicles prepared from bean hypocotyls. Differential sensitivity of ATP-driven proton transport to nitrate, vanadate, N,N′-dicyclohexylcarbodiimide, diethylstilbestrol, and oligomycin suggested the vesicles were enriched for tonoplast. Anion stimulation of proton transport, by enhancement of ATPase activity and dissipation of the membrane potential, was consistent with this conclusion. Although fungal components inhibited the formation of a pH gradient, the membrane potential was unaffected and the ATPase activity slightly stimulated. These data suggest that the fungal components produce an electroneutral proton exchange. Proton transport in Dark Red Kidney bean tonoplast vesicles was inhibited by mycelial preparations from the incompatible α race and compatible β race. Elicitor activity, however, was greater in the α race fractions. Elicitor purified from α race culture filtrate did not inhibit proton transport in vesicles isolated from Dark Red Kidney bean. Consequently, elicitor activity need not be associated with an ability to impair tonoplast function. PMID:16665456

  2. Early membrane events induced by salicylic acid in motor cells of the Mimosa pudica pulvinus.

    Saeedi, Saed; Rocher, Françoise; Bonmort, Janine; Fleurat-Lessard, Pierrette; Roblin, Gabriel


    Salicylic acid (o-hydroxy benzoic acid) (SA) induced a rapid dose-dependent membrane hyperpolarization (within seconds) and a modification of the proton secretion (within minutes) of Mimosa pudica pulvinar cells at concentrations higher than 0.1mM. Observations on plasma membrane vesicles isolated from pulvinar tissues showed that SA acted directly at the membrane level through a protonophore action as suggested by the inhibition of the proton gradient and the lack of effect on H(+)-ATPase catalytic activity. Comparative data obtained with protonophores (carbonylcyanide-m-chlorophenylhydrazone and 2,4-dinitrophenol) and inhibitors of ATPases (vanadate, N,N'-dicyclohexylcarbodiimide, and diethylstilbestrol) corroborated this conclusion. Consequently, the collapse of the proton motive force led to an impairment in membrane functioning. This impairment is illustrated by the inhibition of the ion-driven turgor-mediated seismonastic reaction of the pulvinus following SA treatment. SA acted in a specific manner as its biosynthetic precursor benzoic acid induced much milder effects and the m- and p-OH benzoic acid derivatives did not trigger similar characteristic effects. Therefore, SA may be considered both a membrane signal molecule and a metabolic effector following its uptake in the cells.

  3. Determination of estrogens and estrogenic activities in water from three rivers in Tianjin, China

    Kaifeng Rao; Bingli Lei; Na Li; Mei Ma; Zijian Wang


    Studies on estrogenic disrupting compounds (EDCs) occurrence and identification of main responsible compounds in river water discharged into the sea are of significance.In the present research,we screened estrogenic activities of 10 river water samples from 3 main rivers discharged into Bohai Sea in Tianjin using a recombinant two-hybrid yeast assay and chemical analysis by gas chromatography-mass spectrometry.All sample extracts induced significant estrogenic activity,with 17β-estradiol equivalents (EEQ)of raw water ranging from 5.72 to 59.06 ng/L.Six most concerned EDCs in the river water samples including estrone,17β-estradiol,17α-ethinylestradiol,estriol,diethylstilbestrol and estradiol valerate were determined,with their concentrations up to 50.70,31.40,24.40,37.20,2.56,and 8.47 ng/L,respectively.Through causality analysis by comparing the EEQ values of yeast assay and chemical analysis,17α-ethinylestradiol and 17ββ-estradiol were identified as the main contributors to the estrogenic effects of the river samples,accounting for the whole estrogenic activities (62.99% to 185.66%),and estrogen antagonistic compounds might presented in the heavy polluted water samples.The proposed approach using both chemical analysis and bioassay could be used for identification and evaluation of the estrogenic activity of EDCs in river water.

  4. An estrogen receptor model to describe the regulation of prolactin synthesis by antiestrogens in vitro.

    Lieberman, M E; Gorski, J; Jordan, V C


    A hypothetical model of the ligand interaction with the estrogen receptor binding site has been developed to describe the structural features necessary to initiate or to inhibit prolactin synthesis in vitro. The biological potency of the binding ligands is directly related to their relative binding affinity (RBA) for the estrogen receptor. The relative potencies of antiestrogens to inhibit estradiol-stimulated prolactin synthesis was trans-monohydroxytamoxifen identical to cis-monohydroxytamoxifen identical to tamoxifen, consistent with their RBAs for uterine estrogen receptor. Similarly the relative potency of estrogens to stimulate prolactin synthesis was diethylstilbestrol identical to estradiol greater than ICI 77,949 greater than ICI 47,699 identical to zuclomiphene, consistent with their RBAs. The compound LY126412 (trioxifene without the aminoethoxy side chain) did not interact with the estrogen receptor at the concentrations tested (10(-8)--10(-6) M) or exhibit estrogenic or antiestrogenic properties using the prolactin synthesis assay. Overall, the ligand-receptor model stresses the structural requirement for high affinity binding and the critical positioning of the alkylamino-ethoxy side chain in space (in relation to the ligand-binding site on the estrogen receptor) to prevent prolactin synthesis.

  5. Application of the yeast-based reporter gene bioassay for the assessment of estrogenic activity in cow's milk from Poland.

    Stypuła-Trębas, Sylwia; Minta, Maria; Radko, Lidia; Żmudzki, Jan


    Milk contain compounds acting through the estrogen receptor signaling. The still open question whether such estrogens pose a risk for human health, encouraged us to measure the overall estrogenic activity of cow's milk in the in vitro yeast reporter bioassay. First, we assessed the ability of the bioassay to detect estrogens frequently detected in milk. The relative potencies of 16 compounds descended in the order: 17β-estradiol (17β-E2), 17α-ethinylestradiol, diethylstilbestrol, dienestrol, 17α-E2, estrone, zearalenone, estriol, equol, genistein, 17β-E2 glucuronide, bisphenol A, apigenin, daidzein. Flavone, 4-n-nonylphenol and 4-t-octylphenol shown no activity in the bioassay.The estrogenic activities of milk samples without hydrolysis were below the detection limit, whereas in 50% of the deconjugated samples they varied between 0.29 and 0.49 ng EEQ mL(-1). We also compared the estrogenic activity in raw cow's milk collected from rural and industrial locations in Poland. In our pilot study we did not observe statistically significant difference in estrogenic activities in milk collected from the two locations. We found that the daily intake of estrogens with milk may be higher than estrogen levels in human serum. Further studies are warranted to evaluate the significance of milk and dairy as a source of estrogens for humans.

  6. In vivo imaging of activated estrogen receptors in utero by estrogens and bisphenol A.

    Lemmen, Josephine G; Arends, Roel J; van der Saag, Paul T; van der Burg, Bart


    Environmental estrogens are of particular concern when exposure occurs during embryonic development. Although there are good models to study estrogenic activity of chemicals in adult animals, developmental exposure is much more difficult to test. The weak estrogenic activity of the environmental estrogen bisphenol A (BPA) in embryos is controversial. We have recently generated transgenic mice that carry a reporter construct with estrogen-responsive elements coupled to luciferase. We show that, using this in vivo model in combination with the IVIS imaging system, activation of estrogen receptors (ERs) by maternally applied BPA and other estrogens can be detected in living embryos in utero. Eight hours after exposure to 1 mg/kg BPA, ER transactivation could be significantly induced in the embryos. This was more potent than would be estimated from in vitro assays, although its intrinsic activity is still lower than that of diethylstilbestrol and 17beta-estradiol dipropionate. On the basis of these results, we conclude that the estrogenic potency of BPA estimated using in vitro assays might underestimate its estrogenic potential in embryos.

  7. Crystallization of prostaglandin-H synthase for X-ray structure analysis

    Jahnke, K.; Degen, G.H.; Buehner, M. (Univ. of Wuerzburg (West Germany))


    Prostaglandin-H (PGH) synthase from ram seminal vesicles is a dimeric integral membrane protein of molecular weight 140 kDa. PGH synthase is a key enzyme in the biosynthesis of prostaglandins, has cyclooxygenase and peroxidase activities, and contains heme as a coenzyme. In the peroxidation step of its reaction, PGH synthase can use xenobiotics as co-substrates and can catalyze the metabolic activation of carcinogens such as diethylstilbestrol. To gain a detailed understanding of the inner workings of PGH synthase, the authors are investigating its three-dimensional structure by X-ray crystallography. A purification procedure was established that yields stable homogeneous PGH synthase that is at least 80% holoenzyme. Manipulation of these crystals is very difficult due to the small volume of the growth phase. The crystals dissolved rapidly in all aqueous media into which they were transferred for mounting in X-ray capillaries. Therefore, the authors have not yet been able to demonstrate their true X-ray scattering power. A crystal provisionally dry mounted diffracted to about 8 {angstrom} resolution.

  8. Characteristics of estrogen-induced peroxidase in mouse uterine luminal fluid

    Jellinck, P.H.; Newbold, R.R.; McLachlan, J.A. (Queen' s University, Kingston, Ontario (Canada))


    Peroxidase activity in the uterine luminal fluid of mice treated with diethylstilbestrol was measured by the guaiacol assay and also by the formation of 3H2O from (2-3H)estradiol. In the radiometric assay, the generation of 3H2O and 3H-labeled water-soluble products was dependent on H2O2 (25 to 100 microM), with higher concentrations being inhibitory. Tyrosine or 2,4-dichlorophenol strongly enhanced the reaction catalyzed either by the luminal fluid peroxidase or the enzyme in the CaCl2 extract of the uterus, but decreased the formation of 3H2O from (2-3H)estradiol by lactoperoxidase in the presence of H2O2 (80 microM). NADPH, ascorbate, and cytochrome c inhibited both luminal fluid and uterine tissue peroxidase activity to the same extent, while superoxide dismutase showed a marginal activating effect. Lactoferrin, a major protein component of uterine luminal fluid, was shown not to contribute to its peroxidative activity, and such an effect by prostaglandin synthase was also ruled out. However, it was not possible to exclude eosinophil peroxidase, brought to the uterus after estrogen stimulation, as being the source of peroxidase activity in uterine luminal fluid.

  9. Transferability and inter-laboratory variability assessment of the in vitro bovine oocyte fertilization test.

    Tessaro, Irene; Modina, Silvia C; Crotti, Gabriella; Franciosi, Federica; Colleoni, Silvia; Lodde, Valentina; Galli, Cesare; Lazzari, Giovanna; Luciano, Alberto M


    The dramatic increase in the number of animals required for reproductive toxicity testing imposes the validation of alternative methods to reduce the use of laboratory animals. As we previously demonstrated for in vitro maturation test of bovine oocytes, the present study describes the transferability assessment and the inter-laboratory variability of an in vitro test able to identify chemical effects during the process of bovine oocyte fertilization. Eight chemicals with well-known toxic properties (benzo[a]pyrene, busulfan, cadmium chloride, cycloheximide, diethylstilbestrol, ketoconazole, methylacetoacetate, mifepristone/RU-486) were tested in two well-trained laboratories. The statistical analysis demonstrated no differences in the EC50 values for each chemical in within (inter-runs) and in between-laboratory variability of the proposed test. We therefore conclude that the bovine in vitro fertilization test could advance toward the validation process as alternative in vitro method and become part of an integrated testing strategy in order to predict chemical hazards on mammalian fertility. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. 妇科再造丸对雌孕激素负荷大鼠子宫病理形态学的影响%The Pathomorphological Effects of Uterus in Rats Loaded by Estradiol and Progesterone of Fuke Zaizao Pill

    胡颖; 罗俊; 黄能慧


    目的:研究妇科再造丸(FKW)对雌、孕激素负荷大鼠子宫病理形态学的影响.方法:将60只雌性SD大鼠随机分为6组:正常对照组、模型组、FKW低、中、高(0.28,0.56,1.12g·kg-1·d-1)剂量组、桂枝茯苓胶囊组(0 3 g·kg-1·d-1).除正常对照组外,其余5组肌肉注射己烯雌酚和黄体酮复制大鼠子宫平滑肌瘤样增生模型,从第9周末开始灌胃给予FKW干预,共18周.末次给药后,取大鼠子宫组织HE染色镜下观察其病理形态学改变,同时测定子宫平滑肌厚度.结果:与正常对照组比较,模型组大鼠子宫肌层明显增厚(P<0 01),子宫组织出现肌纤维增生肥大、排列紊乱、穿插及中性粒细胞浸润等病理现象,对上述指标进行半定量统计分析显示差异具有显著性(P<0.01).FKW能显著减少模型大鼠子宫肌层增厚的程度(P <0.05,P <0.01),改善子宫的病理组织形态学变化(P<0.05,P<0.01).结论:FKW可以抑制雌、孕激素负荷引起的大鼠子宫平滑肌瘤样增生.%Objective: To study the pathomorphological change of uterus in rats loaded by estradiol and progesterone and the interfering effects of Feke Zaizao pill( FKW). Method: Sixty female SD rats were divided into 6 groups randomly:the normal group, the model group, the FKW low, moderate, high group(0. 28 ,0. 56,1. 12 g· kg-1·d-1), the Guizhi Fuling capsule group (0. 3 g·kg -1·d-1 ). The rats, except for the normal group, were injected with diethylstilbestrol and progesterone for eighteen weeks to establish uterine leiomyoma-like hyperplasia model. And from the end of the ninth week, FKW was used to interfering with the model rats by gastric perfusion. The uterus tissue was used to processing pathology detection by hematoxylin-eosin staining to observe the pathomorphological changes and the thickness of uterine smooth muscle to study the influence of FKW. Result: Compared with the normal group, the myometrium of rats in the model group

  11. Identification of previously unrecognized antiestrogenic chemicals using a novel virtual screening approach.

    Wang, Ching Y; Ai, Ni; Arora, Sonia; Erenrich, Eric; Nagarajan, Karthigeyan; Zauhar, Randy; Young, Douglas; Welsh, William J


    The physiological roles of estrogen in sexual differentiation and development, female and male reproductive processes, and bone health are complex and diverse. Numerous natural and synthetic chemical compounds, commonly known as endocrine disrupting chemicals (EDCs), have been shown to alter the physiological effects of estrogen in humans and wildlife. As such, these EDCs may cause unanticipated and even undesirable effects. Large-scale in vitro and in vivo screening of chemicals to assess their estrogenic activity would demand a prodigious investment of time, labor, and money and would require animal testing on an unprecedented scale. Approaches in silico are increasingly recognized as playing a vital role in screening and prioritizing chemicals to extend limited resources available for experimental testing. Here, we evaluated a multistep procedure that is suitable for in silico (virtual) screening of large chemical databases to identify compounds exhibiting estrogenic activity. This procedure incorporates Shape Signatures, a novel computational tool that rapidly compares molecules on the basis of similarity in shape, polarity, and other bio-relevant properties. Using 4-hydroxy tamoxifen (4-OH TAM) and diethylstilbestrol (DES) as input queries, we employed this scheme to search a sample database of approximately 200,000 commercially available organic chemicals for matches (hits). Of the eight compounds identified computationally as potentially (anti)estrogenic, biological evaluation confirmed two as heretofore unknown estrogen antagonists. Subsequent radioligand binding assays confirmed that two of these three compounds exhibit antiestrogenic activities comparable to 4-OH TAM. Molecular modeling studies of these ligands docked inside the binding pocket of estrogen receptor alpha (ERalpha) elucidated key ligand-receptor interactions that corroborate these experimental findings. The present study demonstrates the utility of our computational scheme for this and

  12. Radical-scavenging Activity of Estrogen and Estrogen-like Compounds Using the Induction Period Method

    Seiichiro Fujisawa


    Full Text Available The radical-scavenging activity of estrogens (estrone, 2-hydroxyestradiol,estrogen-like compounds (diethylstilbestrol, DES; bisphenol A, BPA and the mono-phenolic compound 2,6-di-t-butyl-4-methoxyphenol (BMP was investigated using themethod of measuring the induction period for polymerization of methyl methacrylate(MMA initiated by thermal decomposition of 2,2'-azobisisobutyronitrile (AIBN andbenzoyl peroxide (BPO at 70°C using differential scanning calorimetry (DSC. Thestoichiometric factor (n, number of free radicals trapped by one mole of antioxidantmoiety for the AIBN system declined in the order BMP (2.0, 2-hydroxyestradiol (2.0>DES (1.3 > BPA (1.2 > estrone (0.9, whereas that for the BPO system declined in theorder BMP (2.0 >DES (1.9, BPA (1.9 > estrone (1.3 > 2-hydroxyestradiol (0.7. Theinhibition rate constant (kinh x 10-3 M-1s-1 for the AIBN system declined in the orderestrone (2.2 > BPA (2.0 > DES (1.9 > 2-hydroxyestradiol (1.2 > BMP (1.1, whereasthat for the BPO system declined in the order 2-hydroxyestradiol (3.2 > estrone (1.4 >DES (1.2 > BPA (1.0 > BMP (0.9. The radical-scavenging activity for bioactivecompounds such as estrogens should be evaluated using these two methods (the n and kinhto elucidate the mechanism of a particular reaction. The great difference of the n and kinhfor estrogens between the AIBN and BPO system suggested that their oxidation process iscomplex.

  13. Resveratrol treatment delays growth plate fusion and improves bone growth in female rabbits.

    Elham Karimian

    Full Text Available Trans-resveratrol (RES, naturally produced by many plants, has a structure similar to synthetic estrogen diethylstilbestrol, but any effect on bone growth has not yet been clarified. Pre-pubertal ovary-intact New Zealand white rabbits received daily oral administration of either vehicle (control or RES (200 mg/kg until growth plate fusion occurred. Bone growth and growth plate size were longitudinally monitored by X-ray imaging, while at the endpoint, bone length was assessed by a digital caliper. In addition, pubertal ovariectomized (OVX rabbits were treated with vehicle, RES or estradiol cypionate (positive control for 7 or 10 weeks and fetal rat metatarsal bones were cultured in vitro with RES (0.03 µM-50 µM and followed for up to 19 days. In ovary-intact rabbits, sixteen-week treatment with RES increased tibiae and vertebrae bone growth and subsequently improved final length. In OVX rabbits, RES delayed fusion of the distal tibia, distal femur and proximal tibia epiphyses and femur length and vertebral bone growth increased when compared with controls. Histomorphometrical analysis showed that RES-treated OVX rabbits had a wider distal femur growth plate, enlarged resting zone, increased number/size of hypertrophic chondrocytes, increased height of the hypertrophic zone, and suppressed chondrocyte expression of VEGF and laminin. In cultured fetal rat metatarsal bones, RES stimulated growth at 0.3 µM while at higher concentrations (10 μM and 50 μM growth was inhibited. We conclude that RES has the potential to improve longitudinal bone growth. The effect was associated with a delay of growth plate fusion resulting in increased final length. These effects were accompanied by a profound suppression of VEGF and laminin expression suggesting that impairment of growth plate vascularization might be an underlying mechanism.

  14. Cellular heterogeneity in the membrana granulosa of developing rat follicles: assessment by flow cytometry and lectin binding.

    Kerketze, K; Blaschuk, O W; Farookhi, R


    The hormone-mediated maturation of ovarian follicles is apparently accompanied by position-specific differentiation of cells of the membrana granulosa. We have assessed the extent of this cellular heterogeneity by flow cytometry using a variety of fluorescein isothiocyanate-labeled lectins as probes. Follicular development was stimulated in immature rats by treatment with either diethylstilbestrol (DES) or equine CG (eCG). Lectin binding to monodispersed rat granulosa cells was then analyzed by flow cytometry. Our results demonstrate that there are two distinct populations of small (4-7 microM) and large (9-12 microM) granulosa cells in follicles from DES- and eCG-treated animals. Both populations appear to be mitotically active and show specific lectin-binding characteristics. Six lectins (canavalia ensiforms, triticum vulgaris, maclura pomifera, erythrina cristagalli, jacalin, and vicia villosa) bind equally to both small and large granulosa cells from the DES- and eCG-treated rats. In contrast, no binding to either cell population was detected with six other lectins (dolichos biflorus, griffonia simplicifolia-II, lycopersicon esculentum, datura stramonium, solanum tuberosum, and ulex europaeus). Furthermore, four galactose-binding lectins (bauhinia purpurea, glysine maximus, griffonia simplicifolia-I, and arachis hypogaea) were found to identify specific subsets of granulosa cells. Three of these lectins (bauhinia purpurea, glysine maximus, and griffonia simplicifolia-I) bind to only small granulosa cells from either DES- or eCG- treated immature rats. The fourth lectin (arachis hypogaea) identifies subpopulations of both small and large granulosa cells. Application of the four galactose-specific lectins to fixed sections of frozen ovaries demonstrated binding to the perioocyte and cumulus granulosa cells. We conclude that cellular heterogeneity exists within the follicular epithelium at various stages-specific lectin-binding sites.

  15. An automated solid-phase microextraction method based on magnetic molecularly imprinted polymer as fiber coating for detection of trace estrogens in milk powder.

    Lan, Hangzhen; Gan, Ning; Pan, Daodong; Hu, Futao; Li, Tianhua; Long, Nengbing; Qiao, Li


    A new automated solid-phase micro extraction (SPME) sampling method was developed for quantitative enrichment of estrogens (ES) from milk powder, using magnetic molecularly imprinted polymer (MMIP) as fiber coating. The method (MMIP-SPME) was built with several electromagnetic stainless steel fibers, placed in parallel for simultaneously extraction. The MMIP was synthesized using core-shell Fe3O4@SiO2 nanoparticles (NPs) as magnetic support. Estradiol (E2) was employed as the template molecule, acrylamide (AA) as functional monomer, and ethylene glycol dimethacrylate (EGDMA) as cross-linker. MMIP can be easily absorbed or desorbed from fibers when the current is turned on or off, creating magnetism. Compared to traditional MIP-SPME, the prepared procedure of MMIP-SPME is time-saving and organic solvent-free. The proposed device significantly improved the efficiency of separation and enrichment of estrogens from complex matrices thereby and facilitating the pretreatment steps by electromagnetically controlled extraction fibers to achieve full automation. Several experimental parameters were studied, including extraction and desorption kinetics, solution pH, desorption solution, ratio, and shuttle rate. The newly developed MMIP-SPME showed good sensitivity and high binding capacity, fast adsorption kinetics and desorption kinetics for estrone (E1), estradiol (E2), estriol (E3) and diethylstilbestrol (DES) under optimized conditions. The detection limits for the four estrogens were 1.5-5.5ngg(-1) with excellent reproducibility (RSD values less than 7.1%) when milk powder samples spiked with analytes at 20, 100 and 250ngg(-1) were studied.

  16. Synthesis and biological evaluation of a fluorine-18-labeled nonsteroidal androgen receptor antagonist, N-(3-[{sup 18}F]fluoro-4-nitronaphthyl)-cis-5-norbornene-endo-2,3-dicarboxylic imide

    Parent, Ephraim E. [Department of Chemistry, University of Illinois, Urbana, IL 61801 (United States); Dence, Carmen S. [Washington University School of Medicine, St. Louis, MO 63110 (United States); Sharp, Terry L. [Washington University School of Medicine, St. Louis, MO 63110 (United States); Welch, Michael J. [Washington University School of Medicine, St. Louis, MO 63110 (United States); Katzenellenbogen, John A. [Department of Chemistry, University of Illinois, Urbana, IL 61801 (United States)]. E-mail:


    Introduction: Androgen receptor (AR), which is overexpressed in most prostate cancers, is the target of androgen ablation and antiandrogen therapies: it is also the target for the receptor-mediated imaging of AR-positive prostate cancer using radiolabeled ligands. Previous AR imaging agents were based on a steroidal core labeled with fluorine. To develop a novel class of nonsteroidal imaging agents, with binding and pharmacological characteristics that are more similar to those of clinically used AR antagonists, we synthesized N-(3-fluoro-4-nitronaphthyl)-cis-5-norbornene-endo-2,3-dicarboxylic imide (3-F-NNDI), an analog of recently reported AR antagonist ligands. Methods: 3-F-NNDI was synthesized in six steps starting with 1-nitronaphthalene, with fluorine incorporation as the final step. The labeling of 3-F-NNDI with fluorine-18 was achieved through a novel, extremely mild, S{sub N}Ar displacement reaction of an o-nitro-activated arene trimethylammonium salt, and 3-[{sup 18}F]F-NNDI was prepared in high specific activity. Results and Discussion: 3-F-NNDI was found to have an AR-binding affinity similar to that of its parent compound. In vitro assays demonstrated high stability of the labeled compound under physiological conditions in buffer and in the blood. Androgen target tissue uptake in diethylstilbestrol-pretreated male rats, however, was minimal, probably because of extensive metabolic defluorination the radiolabeled ligand. Conclusions: This study is part of our first look at a novel class of nonsteroidal AR antagonists as positron emission tomography (PET) imaging agents that are alternatives to steroidal AR agonist-based imaging agents. Although 3-[{sup 18}F]F-NNDI has significant affinity for AR, it showed limited promise as a PET imaging agent because of its poor target tissue distribution properties.

  17. Polydimethylsiloxane/metal-organic frameworks coated stir bar sorptive extraction coupled to high performance liquid chromatography-ultraviolet detector for the determination of estrogens in environmental water samples.

    Hu, Cong; He, Man; Chen, Beibei; Zhong, Cheng; Hu, Bin


    In this work, three kinds of metal-organic frameworks (MOFs), MOF-5, MOF-199 and IRMOF-3, were introduced in stir bar sorptive extraction (SBSE) and novel polydimethylsiloxane (PDMS)/MOFs (including PDMS/MOF-5, PDMS/MOF-199 and PDMS/IRMOF-3) coated stir bars were prepared by sol-gel technique. These PDMS/MOFs coatings were characterized and critically compared for the extraction of seven target estrogens (17-β-estradiol, dienestrol, diethylstilbestrol, estrone, 4-t-octylphenol, bisphenol-A and 17α-ethynylestradiol) by SBSE, and the results showed that PDMS/IRMOF-3 exhibited highest extraction efficiency. Based on the above facts, a novel method of PDMS/IRMOF-3 coating SBSE-high performance liquid chromatography ultraviolet (HPLC-UV) detection was developed for the determination of seven target estrogens in environmental waters. Several parameters affecting extraction of seven target estrogens by SBSE (PDMS/IRMOF-3) including extraction time, stirring rate, pH, ionic strength, desorption solvent and desorption time were investigated. Under the optimal experimental conditions, the limits of detection (LODs, S/N=3) were found to be in the range of 0.15-0.35 μg/L. The linear range was 2-2,500 μg/L for 17α-ethynylestradiol and 1-2,500 μg/L for other estrogens. The relative standard deviations (RSDs) were in the range of 3.7-9.9% (n=8, c=20 μg/L) and the enrichment factors were from 30.3 to 55.6-fold (theoretical enrichment factor was 100-fold). The proposed method was successfully applied to the analysis of estrogens in environmental water samples, and quantitative recoveries were obtained for the spiking experiments.

  18. Oxytocin and its receptors are synthesized in the rat vasculature.

    Jankowski, M; Wang, D; Hajjar, F; Mukaddam-Daher, S; McCann, S M; Gutkowska, J


    Produced and released by the heart, oxytocin (OT) acts on its cardiac receptors to decrease the cardiac rate and force of contraction. We hypothesized that it might also be produced in the vasculature and regulate vascular tone. Consequently, we prepared acid extracts of the pulmonary artery and vena cava of female rats. OT concentrations in dog and sheep aortae were equivalent to those of rat aorta (2745 +/- 180 pg/mg protein), indicating that it is present in the vasculature of several mammalian species. Reverse-phase HPLC of aorta and vena cava extracts revealed a single peak corresponding to the amidated OT nonapeptide. Reverse-transcribed PCR confirmed OT synthesis in these tissues. Using the selective OT receptor ligand compound VI, we detected a high number of OT-binding sites in the rat vena cava and aorta. Furthermore, OT receptor (OTR) mRNA was found in the vena cava, pulmonary vein, and pulmonary artery with lower levels in the aorta, suggesting vessel-specific OTR distribution. The abundance of OTR mRNA in the vena cava and pulmonary vein was associated with high atrial natriuretic peptide mRNA. In addition, we have demonstrated that diethylstilbestrol treatment of immature female rats increased OT significantly in the vena cava but not in the aorta and augmented OTR mRNA in both the aorta (4-fold) and vena cava (2-fold), implying regulation by estrogen. Altogether, these data suggest that the vasculature contains an intrinsic OT system, which may be involved in the regulation of vascular tone as well as vascular regrowth and remodeling.

  19. Nuclear receptors and endocrine disruptors in fetal and neonatal testes: a gapped landscape.

    Virginie eRouiller-Fabre


    Full Text Available During the last decades, many studies reported that male reproductive disorders are increasing among humans. It is currently acknowledged that these abnormalities can result from fetal exposure to environmental chemicals that are progressively becoming more concentrated and widespread in our environment. Among the chemicals present in the environment (air, water, food and many consumer products, several can act as endocrine disrupting compounds (EDCs, thus interfering with the endocrine system. Phthalates, bisphenol A (BPA and diethylstilbestrol (DES have been largely incriminated, particularly during the fetal and neonatal period, due to their estrogenic and/or anti-androgenic properties. Indeed, many epidemiological and experimental studies have highlighted their deleterious impact on fetal and neonatal testis development. As EDCs can affect many different genomic and non-genomic pathways, the mechanisms underlying the adverse effects of EDC exposure are difficult to elucidate. Using literature data and results from our laboratory, in the present review we discuss the role of classical nuclear receptors (genomic pathway in the fetal and neonatal testis response to EDC exposure, particularly to phthalates, BPA and DES. Among the nuclear receptors we focused on some of the most likely candidates, such as peroxisome-proliferator activated receptor (PPAR, androgen receptor (AR, estrogen receptors (ERα and β, liver X receptors (LXR and small heterodimer partner (SHP. First, we describe the expression and potential functions (based on data from studies using receptor agonists and mouse knockout models of these nuclear receptors in the developing testis. Then, for each EDC studied, we summarize the main evidences indicating that the reprotoxic effect of each EDC under study is mediated through a specific nuclear receptor(s. We also point-out the involvement of other receptors and nuclear receptor-independent pathways.

  20. Postweaning Exposure to Dietary Zearalenone, a Mycotoxin, Promotes Premature Onset of Puberty and Disrupts Early Pregnancy Events in Female Mice

    Ye, Xiaoqin


    Zearalenone (ZEA) is a mycotoxin commonly found in contaminated livestock feed and human food with levels in the range of ppb and low ppm. It was hypothesized that ZEA, an endocrine disruptor, could affect puberty and early pregnancy. To test this hypothesis, newly weaned (3 weeks old) C57BL/6J female mice were exposed to 0, 0.002, 4, 10, and 40 ppm ZEA and 0.05 ppm diethylstilbestrol (positive control) in phytoestrogen-free AIN-93G diet. Females exposed to 10 and 40 ppm ZEA diets showed earlier onset of vaginal opening. Those treated with 40 ppm ZEA diet also had earlier first copulation plug and irregular estrous cyclicity. At 8 weeks old, all females were mated with untreated stud males on AIN-93G diet during mating. Treatment resumed upon identification of a vaginal plug on gestation day 0.5 (D0.5). Embryo implantation was assessed on D4.5. Exposure to 40 ppm ZEA diet resulted in reduced percentage of plugged mice with implantation sites, distended uterine appearance, and retained expression of progesterone receptor in D4.5 uterine epithelium. To determine the exposure timing and mechanisms of disrupted embryo implantation, four groups of females were fed with 0 or 40 ppm ZEA diets during premating (weaning to mating) and postmating (D0.5–D4.5), respectively. Premating exposure to 40 ppm ZEA diet reduced fertilization rate, whereas postmating exposure to 40 ppm ZEA diet delayed embryo transport and preimplantation embryo development, which subsequently affected embryo implantation. These data demonstrate that postweaning exposure to dietary ZEA can promote premature onset of puberty and disrupt early pregnancy events. PMID:23291560

  1. Determination of 13 endocrine disrupting chemicals in sediments by gas chromatography-mass spectrometry using subcritical water extraction coupled with dispersed liquid-liquid microextraction and derivatization.

    Yuan, Ke; Kang, Haining; Yue, Zhenfeng; Yang, Lihua; Lin, Li; Wang, Xiaowei; Luan, Tiangang


    In this study, a sample pretreatment method was developed for the determination of 13 endocrine disrupting chemicals (EDCs) in sediment samples based on the combination of subcritical water extraction (SWE) and dispersed liquid-liquid microextraction (DLLME). The subcritical water that provided by accelerated solvent extractor (ASE) was the sample solution (water) for the following DLLME and the soluble organic modifier that spiked in the subcritical water was also used as the disperser solvent for DLLME in succession. Thus, several important parameters that affected both SWE and DLLME were investigated, such as the extraction solvent for DLLME (chlorobenzene), extraction time for DLLME (30s), selection of organic modifier for SWE (acetone), volume of organic modifier (10%) and extraction temperature for SWE (150 °C). In addition, good chromatographic behavior was achieved for GC-MS after derivatisation by using N,O-bis(trimethylsilyl) trifluoroacetamide (BSTFA). As a result, proposed method sensitive and reliable with the limits of detection (LODs) ranging from 0.006 ng g(-1) (BPA) to 0.639 ng g(-1) (19-norethisterone) and the relative standard deviations (RSDs) between 1.5% (E2) and 15.0% (DES). Moreover, the proposed method was compared with direct ASE extraction that reported previously, and the results showed that SWE-DLLME was more promising with recoveries ranging from 42.3% (dienestrol) to 131.3% (4,5α-dihydrotestosterone), except for diethylstilbestrol (15.0%) and nonylphenols (29.8%). The proposed method was then successfully applied to determine 13 EDCs sediment of Humen outlet of the Pearl River, 12 of target compounds could be detected, and 10 could be quantitative analysis with the total concentration being 39.6 ng g(-1), and which indicated that the sediment of Humen outlet was heavily contaminated by EDCs.

  2. Regulation and dysregulation of vitellogenin mRNA accumulation in daphnids (Daphnia magna)

    Hannas, Bethany R.; Wang, Ying H.; Thomson, Susanne; Kwon, Gwijun; Li Hong [Department of Environmental and Molecular Toxicology, North Carolina State University, Raleigh, NC 27695-7633 (United States); LeBlanc, Gerald A., E-mail: [Department of Environmental and Molecular Toxicology, North Carolina State University, Raleigh, NC 27695-7633 (United States)


    The induction of vitellogenin in oviparous vertebrates has become the gold standard biomarker of exposure to estrogenic chemicals in the environment. This biomarker of estrogen exposure also has been used in arthropods, however, little is known of the factors that regulate the expression of vitellogenin in these organisms. We investigated changes in accumulation of mRNA products of the vitellogenin gene Vtg2 in daphnids (Daphnia magna) exposed to a diverse array of chemicals. We further evaluated the involvement of hormonal factors in the regulation of vitellogenin expression that may be targets of xenobiotic chemicals. Expression of the Vtg2 gene was highly responsive to exposure to various chemicals with an expression range spanning approximately four orders of magnitude. Chemicals causing the greatest induction were piperonyl butoxide, chlordane, 4-nonylphenol, cadmium, and chloroform. Among these, only 4-nonylphenol is recognized to be estrogenic. Exposure to several chemicals also suppressed Vtg2 mRNA levels, as much as 100-fold. Suppressive chemicals included cyproterone acetate, acetone, triclosan, and atrazine. Exposure to the estrogens diethylstilbestrol and bisphenol A had little effect on vitellogenin mRNA levels further substantiating that these genes are not induced by estrogen exposure. Exposure to the potent ecdysteroids 20-hydroxyecdysone and ponasterone A revealed that Vtg2 was subject to strong suppressive control by these hormones. Vtg2 mRNA levels were not significantly affected from exposure to several juvenoid hormones. Results indicate that ecdysteroids are suppressors of vitellogenin gene expression and that vitellogenin mRNA levels can be elevated or suppressed in daphnids by xenobiotics that elicit antiecdysteroidal or ecdysteroidal activity, respectively. Importantly, daphnid Vtg2 is not elevated in response to estrogenic activity.

  3. Determination of three estrogens in cheese by Solid-Phase Dispersive Extraction Coupled with High Performance Liquid Chromatography%固相分散萃取-液相色谱法测定奶酪中3种雌激素

    周建科; 唐翠苓; 韩朝家; 徐鹏


    建立了同时测定奶酪中炔雌醇、己烯雌酚和双烯雌酚三种雌激素的高效液相色谱分析方法.采用固相分散萃取技术处理样品,弗罗里硅土为分散剂、乙腈为萃取剂,经氮气流浓缩,反相色谱分离,紫外230 nm检测得出结果.3种激素回收率分别为89.16%,98.22%和83.22%,相对标准偏差2.98%,1.20%和2.75%,方法检出限分别为0.25,0.25和0.12 μg/g.对市售奶酪进行了实际测定,三种雌激素均未检出.%An efficient method for analyzing three estrogens (ethinyloestradiol , diethylstilbestrol; dienestrol) in cheese by high performance liquid chromatography was developed. The solid-phase dispersion extraction technology was used for sample treatment The dispersant was Florisil, and the extractant was acetonitrile .The extract was concentrated by nitrogen flow, three estrogens were separated and detected by the reverse chromatography at uv 230 nm to obtain the result. Three estrogens recoveries are 89.16%, 98.22% and 83.22%, respectively. Relative standard deviations are 2.98%,1.20% and 2.75%, the detection limit are 0.25 μg/g, 0.25 μg/g and 0.12 μg/g, respectively. The sold cheese is determined, and the three kinds of estrogen is not detected in here.

  4. In utero bisphenol A exposure disrupts germ cell nest breakdown and reduces fertility with age in the mouse

    Wang, Wei, E-mail:; Hafner, Katlyn S., E-mail:; Flaws, Jodi A., E-mail:


    Bisphenol A (BPA) is a known reproductive toxicant in rodents. However, the effects of in utero BPA exposure on early ovarian development and the consequences of such exposure on female reproduction in later reproductive life are unclear. Thus, we determined the effects of in utero BPA exposure during a critical developmental window on germ cell nest breakdown, a process required for establishment of the finite primordial follicle pool, and on female reproduction. Pregnant FVB mice (F0) were orally dosed daily with tocopherol-striped corn oil (vehicle), diethylstilbestrol (DES; 0.05 μg/kg, positive control), or BPA (0.5, 20, and 50 μg/kg) from gestational day 11 until birth. Ovarian morphology and gene expression profiles then were examined in F1 female offspring on postnatal day (PND) 4 and estrous cyclicity was examined daily after weaning for 30 days. F1 females were also subjected to breeding studies with untreated males at three to nine months. The results indicate that BPA inhibits germ cell nest breakdown via altering expression of selected apoptotic factors. BPA also significantly advances the age of first estrus, shortens the time that the females remain in estrus, and increases the time that the females remain in metestrus and diestrus compared to controls. Further, F1 females exposed to low doses of BPA exhibit various fertility problems and have a significantly higher percentage of dead pups compared to controls. These results indicate that in utero exposure to low doses of BPA during a critical ovarian developmental window interferes with early ovarian development and reduces fertility with age. - Highlights: • In utero BPA exposure inhibits germ cell nest breakdown in female mouse offspring. • In utero BPA exposure alters expression of apoptosis regulators in the ovaries of mouse offspring. • In utero BPA exposure advances first estrus age and alters cyclicity in mouse offspring. • In utero BPA exposure causes various fertility problems in

  5. Reproductive and developmental effects of endocrine disrupters in invertebrates: in vitro and in vivo approaches.

    Hutchinson, Thomas H


    In order to gain basic understanding in the ecotoxicity of endocrine disrupting chemicals or EDCs (including natural chemicals and some pharmaceuticals), many international research groups are currently testing these chemicals using aquatic invertebrates. This paper discusses relevant examples to address key questions: which aquatic invertebrates are likely to be vulnerable to mammalian and non-mammalian EDCs; and which types of invertebrate chronic tests might be most sensitive and cost-effective to address potential environmental exposures? For a full review of invertebrate endocrine disrupter research see Endocrine Disruption in Invertebrates: Endocrinology, Testing and Assessment (1999). As an example, crustaceans are a particular focus of EDC research, reflecting their abundance in nature, commercial importance and their inclusion in the regulatory assessment schemes for active pharmaceutical ingredients (APIs). There is a diverse literature on the developmental and reproductive effects of mammalian EDCs in Crustacea, although there is growing evidence that such effects are probably not mediated via arthropod hormone systems. For example, recent studies in Europe using a marine copepod (Tisbe battagliai) life-cycle test have evaluated ecdysteroid agonists (e.g. 20-hydroxyecdysone), oestrogen agonists (e.g. diethylstilbestrol (DES), 17beta-oestradiol, oestrone and 17alpha-ethynylestradiol) and the pharmaceutical anti-oestrogen (ZM189, 154). While 20-hydroxyecdysone and DES were highly toxic, the other compounds tested show no significant toxicity to copepods. Furthermore, in vitro studies indicate that these environmental EDCs and several related APIs are not active against the ecdysteroid receptor. Therefore, other undefined modes of action appear to be responsible for crustacean toxicity in vivo and caution should be exercised before ascribing any apical effects to endogenous endocrine mechanisms, or before crustacean "EDC" data are extrapolated to other

  6. Nuclear receptors and endocrine disruptors in fetal and neonatal testes: a gapped landscape.

    Rouiller-Fabre, Virginie; Guerquin, Marie Justine; N'Tumba-Byn, Thierry; Muczynski, Vincent; Moison, Delphine; Tourpin, Sophie; Messiaen, Sébastien; Habert, René; Livera, Gabriel


    During the last decades, many studies reported that male reproductive disorders are increasing among humans. It is currently acknowledged that these abnormalities can result from fetal exposure to environmental chemicals that are progressively becoming more concentrated and widespread in our environment. Among the chemicals present in the environment (air, water, food, and many consumer products), several can act as endocrine disrupting compounds (EDCs), thus interfering with the endocrine system. Phthalates, bisphenol A (BPA), and diethylstilbestrol (DES) have been largely incriminated, particularly during the fetal and neonatal period, due to their estrogenic and/or anti-androgenic properties. Indeed, many epidemiological and experimental studies have highlighted their deleterious impact on fetal and neonatal testis development. As EDCs can affect many different genomic and non-genomic pathways, the mechanisms underlying the adverse effects of EDC exposure are difficult to elucidate. Using literature data and results from our laboratory, in the present review, we discuss the role of classical nuclear receptors (genomic pathway) in the fetal and neonatal testis response to EDC exposure, particularly to phthalates, BPA, and DES. Among the nuclear receptors, we focused on some of the most likely candidates, such as peroxisome-proliferator activated receptor (PPAR), androgen receptor (AR), estrogen receptors (ERα and β), liver X receptors (LXR), and small heterodimer partner (SHP). First, we describe the expression and potential functions (based on data from studies using receptor agonists and mouse knockout models) of these nuclear receptors in the developing testis. Then, for each EDC studied, we summarize the main evidences indicating that the reprotoxic effect of each EDC under study is mediated through a specific nuclear receptor(s). We also point-out the involvement of other receptors and nuclear receptor-independent pathways.

  7. Establishment of a transgenic yeast screening system for estrogenicity and identification of the anti-estrogenic activity of malachite green.

    Jiao, Baowei; Yeung, Eric K C; Chan, Chi Bun; Cheng, Christopher H K


    Endocrine disruptors refer to chemical compounds in the environment which interfere with the endocrine systems of organisms. Among them, environmental estrogens pose serious problems to aquatic organisms, in particular fish. It is therefore important and necessary to have a fast and low-cost system to screen the large number of different chemical compounds in the aquatic environment for their potential endocrine disrupting actions. In this study, a screening platform was developed to detect xenoestrogens in the aquatic environment using the fission yeast Schizosaccharomyces pombe, and applied for compound screening. The aim was to demonstrate any significant potential differences between the fish screening system and the human screening system. To this end, a yeast expression vector harboring a fish estrogen receptor alpha and a reporter vector containing the estrogen responsive element fused with the Escherichia coli LacZ gene were constructed. After transformation with these two vectors, the transformed yeast clones were confirmed by Western blotting and selected on the basis of the beta-galactosidase activity. In this transgenic yeast system, the natural estrogen (estradiol) and other known xenoestrogens such as diethylstilbestrol, bisphenol A, genistein and dichloro-diphenyl-trichloroethane exhibited dose-dependent activities. Using this system, more than 40 putative endocrine disruptors including phytoestrogens, pesticides, herbicides, industrial dyes and other industrial chemicals were screened. Ten of them were demonstrated to exhibit estrogenic actions. Industrial dyes such as malachite green (MG) that disrupt thyroid hormone synthesis are extensively used and are widely distributed in the aquatic environment. Using this system, MG did not show any estrogenic action, but was demonstrated to exhibit anti-estrogenic activity.

  8. Polyacetal-stilbene conjugates - The first examples of polymer therapeutics for the inhibition of HIF-1 in the treatment of solid tumours.

    England, Richard M; Masiá, Esther; Giménez, Vanessa; Lucas, Rut; Vicent, María J


    We report here the first examples of Polymer Therapeutics synthesised with the intention of inhibiting Hypoxia Inducible Factor-1 (HIF-1), a transcription factor heavily involved in numerous cell processes under a low oxygen environment. Four compounds were selected for use in these systems; Diethylstilbestrol (DES), Bisphenol A (BIS), Dienestrol (DIENES) and Hexestrol (HEX), which were chosen from a large family of similar molecules known as Stilbenes. These are non-steroidal molecules with structural similarities to oestrogen, and of which DES and BIS have previously been reported for HIF-1 inhibition. These molecules were incorporated into a poly(ethylene glycol) (PEG) based polyacetal system using a reaction of short PEG chains with di(ethylene glycol) divinyl ether units and an acid catalyst and without the need for biodegradable linkers. With an improved polyacetal synthesis strategy we obtained high yields of water soluble polymer conjugates with desirable drug loadings and tailored molecular weights (Mw 23,000-35,000g/mol) with relatively narrow polydispersities (pdi 1.3-1.5). These polymers were found to be hydrolytically cleaved under acid conditions (such as those found in endosomes, lysosomes or the extracellular fluid of some tumours) yielding the free drug. Additionally, they were found to be stable over prolonged periods of time at pH 7.4 mimicking blood plasma. Of the four polymers synthesised, the conjugates of DES and BIS displayed the best activity for HIF-1α inhibition in HeLa 9xHRE-Luc tumour cells. More importantly, these conjugates were found to exhibit little to no cell toxicity, contrary to the free drugs, and consequently, they significantly enhanced drug therapeutic index (TI 3.5 vs. 7.2 for free DES vs. DES-polyacetal 2a, and TI 1.1 vs. >20 for free BIS vs. BIS-polyacetal 1b).

  9. Somatic mutations in stilbene estrogen-induced Syrian hamster kidney tumors identified by DNA fingerprinting

    Roy Deodutta


    Full Text Available Abstract Kidney tumors from stilbene estrogen (diethylstilbestrol-treated Syrian hamsters were screened for somatic genetic alterations by Random Amplified Polymorphic DNA-polymerase chain-reaction (RAPD-PCR fingerprinting. Fingerprints from tumor tissue were generated by single arbitrary primers and compared with fingerprints for normal tissue from the same animal, as well as normal and tumor tissues from different animals. Sixty one of the arbitrary primers amplified 365 loci that contain approximately 476 kbp of the hamster genome. Among these amplified DNA fragments, 44 loci exhibited either qualitative or quantitative differences between the tumor tissues and normal kidney tissues. RAPD-PCR loci showing decreased and increased intensities in tumor tissue DNA relative to control DNA indicate that loci have undergone allelic losses and gains, respectively, in the stilbene estrogen-induced tumor cell genome. The presence or absence of the amplified DNA fragments indicate homozygous insertions or deletions in the kidney tumor DNA compared to the age-matched normal kidney tissue DNA. Seven of 44 mutated loci also were present in the kidney tissues adjacent to tumors (free of macroscopic tumors. The presence of mutated loci in uninvolved (non-tumor surrounding tissue adjacent to tumors from stilbene estrogen-treated hamsters suggests that these mutations occurred in the early stages of carcinogenesis. The cloning and sequencing of RAPD amplified loci revealed that one mutated locus had significant sequence similarity with the hamster Cyp1A1 gene. The results show the ability of RAPD-PCR to detect and isolate, in a single step, DNA sequences representing genetic alterations in stilbene estrogen-induced cancer cells, including losses of heterozygosity, and homozygous deletion and insertion mutations. RAPD-PCR provides an alternative molecular approach for studying cancer cytogenetics in stilbene estrogen-induced tumors in humans and experimental

  10. Impact of estrogenic compounds on DNA integrity in human spermatozoa: Evidence for cross-linking and redox cycling activities

    Bennetts, L.E.; De Iuliis, G.N.; Nixon, B.; Kime, M.; Zelski, K. [ARC Centre of Excellence in Biotechnology and Development and Discipline of Biological Sciences, University of Newcastle, NSW (Australia); McVicar, C.M.; Lewis, S.E. [Obstetrics and Gynaecology, Queen' s University, Belfast (United Kingdom); Aitken, R.J. [ARC Centre of Excellence in Biotechnology and Development and Discipline of Biological Sciences, University of Newcastle, NSW (Australia)], E-mail:


    A great deal of circumstantial evidence has linked DNA damage in human spermatozoa with adverse reproductive outcomes including reduced fertility and high rates of miscarriage. Although oxidative stress is thought to make a significant contribution to DNA damage in the male germ line, the factors responsible for creating this stress have not been elucidated. One group of compounds that are thought to be active in this context are the estrogens, either generated as a result of the endogenous metabolism of androgens within the male reproductive tract or gaining access to the latter as a consequence of environmental exposure. In this study, a wide variety of estrogenic compounds were assessed for their direct effects on human spermatozoa in vitro. DNA integrity was assessed using the Comet and TUNEL assays, lesion frequencies were quantified by QPCR using targets within the mitochondrial and nuclear ({beta}-globin) genomes, DNA adducts were characterized by mass spectrometry and redox activity was monitored using dihydroethidium (DHE) as the probe. Of the estrogenic and estrogen analogue compounds evaluated, catechol estrogens, quercetin, diethylstilbestrol and pyrocatechol stimulated intense redox activity while genistein was only active at the highest doses tested. Other estrogens and estrogen analogues, such as 17{beta}-estradiol, nonylphenol, bisphenol A and 2,3-dihydroxynaphthalene were inactive. Estrogen-induced redox activity was associated with a dramatic loss of motility and, in the case of 2-hydroxyestradiol, the induction of significant DNA fragmentation. Mass spectrometry also indicated that catechol estrogens were capable of forming dimers that can cross-link the densely packed DNA strands in sperm chromatin, impairing nuclear decondensation. These results highlight the potential importance of estrogenic compounds in creating oxidative stress and DNA damage in the male germ line and suggest that further exploration of these compounds in the aetiology of

  11. Determination of endocrine disrupting compounds in fish liver, brain, and muscle using focused ultrasound solid-liquid extraction and dispersive solid phase extraction as clean-up strategy.

    Ros, Oihana; Vallejo, Asier; Olivares, Maitane; Etxebarria, Nestor; Prieto, Ailette


    This study describes a new method for the simultaneous extraction of several endocrine disrupting compounds, including alkylphenols (APs), estrogen, bisphenol-A (BPA) and one phthalate metabolite (mono-2-ethylhexyl ester, MEHP) in fish liver, brain, and muscle. Parameters affecting the extraction (extraction solvent and temperature) and the clean-up (dispersive phase nature and amount) steps were evaluated. The extraction was performed by means of focused ultrasound solid-liquid extraction (FUSLE) using 10 mL of n-hexane:acetone (50:50, v/v) for 5 min at ~0 °C, and the clean-up was done by means of dispersive solid phase extraction (dSPE) using 100 mg of ENVI-CARB and 100 mg of MgSO4 for the cleaning of brain and muscle extracts together with 100 mg of PSA in the case of liver extracts. Good apparent recoveries were obtained in the case of liver (62-132 %), brain (66-120 %), and muscle (74-129 %), relative standard deviation (RSD%) was always below 26 %, and the method detection limits (MDLs) were at low ng/g level. The developed method was applied to fish captured in Urdaibai estuary (Bay of Biscay) in December 2015, and the concentrations obtained were in the range MDL-1115 ng/g in brain, MDL-962 ng/g in muscle, and MDL-672 ng/g in liver. In general, the highest concentrations were measured in liver, followed by brain and muscle. In addition, diethylstilbestrol was only detected in fish brain. Graphical Abstract MS method scheme for the/MS method scheme for the determination of EDCs in fish liver, brain and muscle.

  12. Study and comparison of polydopamine and its derived carbon decorated nanoparticles in the magnetic solid-phase extraction of estrogens.

    Huang, Zhenzhen; Lee, Hian Kee


    Surface functionalization enabled by bioinspired polydopamine (PDA) is recognized as a convenient route for fabrication of multifunctional nanoparticles. In the present work, magnetic nanoparticles with polymer (Fe3O4@PDA) and carbon shell (Fe3O4@C) were prepared by self-oxidation of dopamine, and carbonization of the PDA coating. The performance of the two magnetic sorbents in the extraction and determination of four estrogens, estrone (E1), estradiol (E2), estriol (E3) and diethylstilbestrol (DES) from water samples in the form of magnetic solid-phase extraction was investigated. Orthogonal array design was utilized to facilitate the optimization of the proposed sample preparation approach. The highest extraction capabilities of the two sorbents were achieved under different experimental conditions. Fe3O4@PDA was shown to be superior to Fe3O4@C in the enrichment of estrogens, suggesting stronger interactions were established between the PDA coating and the target compounds. The extraction and desorption operations were enabled more conveniently by magnetic separation and the extracts were analyzed by high-performance liquid chromatography coupled with ultraviolet and fluorescence detection. The limits of detection achieved in the proposed method were in the range of 0.072-0.15ng/mL for E1 and DES, and 0.0017-0.0062ng/mL for E2 and E3. Good precision (>0.9995) was obtained with the linearity ranging from 0.2 to 100ng/mL, and from 0.01 to 5ng/mL. The method developed was assessed by analysis of the estrogens in tap water, drain water and bottled mineral water samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. The effects of in utero bisphenol A exposure on reproductive capacity in several generations of mice

    Ziv-Gal, Ayelet; Wang, Wei; Zhou, Changqing; Flaws, Jodi A.


    In utero bisphenol A (BPA) exposure affects reproductive function in the first generation (F1) of mice; however, not many studies have examined the reproductive effects of BPA exposure on subsequent generations. In this study, pregnant mice (F0) were orally dosed with vehicle, BPA (0.5, 20, and 50 µg/kg/day) or diethylstilbestrol (DES; 0.05 µg/kg/day) daily from gestation day 11 until birth. F1 females were used to generate the F2 generation, and F2 females were used to generate the F3 generation. Breeding studies at the ages of 3, 6, and 9 months were conducted to evaluate reproductive capacity over time. Further, studies were conducted to evaluate pubertal onset, litter size, and percentage of dead pups; and to calculate pregnancy rate, and mating, fertility, and gestational indices. The results indicate that BPA exposure (0.5 and 50 μg/kg/day) significantly delayed the age at vaginal opening in the F3 generation compared to vehicle control. Both DES (0.05 μg/kg/day) and BPA (50 μg/kg/day) significantly delayed the age at first estrus in the F3 generation compared to vehicle control. BPA exposure reduced gestational index in the F1 and F2 generations compared to control. Further, BPA exposure (0.5 μg/kg/day) compromised the fertility index in the F3 generation compared to control. Finally, in utero BPA exposure reduced the ability of female mice to maintain pregnancies as they aged. Collectively, these data suggest that BPA exposure affects reproductive function in female mice and that some effects may be transgenerational in nature. PMID:25771130

  14. Priapismo na doença falciforme Sickle cell priapism

    Perla Vicari


    Full Text Available Priapismo é a complicação relativamente freqüente na doença falciforme. Consiste de ereção peniana prolongada e dolorosa, não acompanhada de desejo ou estímulo sexual, usualmente persistente por mais de quatro horas. A disfunção erétil é seqüela comum no tratamento inadequado. A forma típica de priapismo nestes pacientes é a de baixo fluxo, ocorrendo, ainda, a forma de priapismo recorrente ou stuttering. O tratamento inicial para esta complicação ainda não está bem estabelecido. São propostas várias opções medicamentosas, tais como agonistas adrenérgicos, hormônios análogos à gonadotrofina, dietil-estilbestrol, hidroxiuréia entre outras. Nos casos de falha com as medidas conservadoras e medicamentosas, a intervenção cirúrgica, com confecção de shunt cavernoso, é necessária. Este estudo revisa a incidência, patogênese e opções terapêuticas desta complicação na doença falciforme.Priapism is a common complication of sickle cell disease. It is defined as a painful and persistent penil erection not accompanied by sexual desire or stimulation, usually lasting for more than 4 hours. The typical forms of priapism in sickle cell disease are low-flow and recurrent priapism (stuttering. The first-line treatment for this complication is not totally clear. Several treatments have been proposed such as adrenergic agents, gonadotropin-releasing hormone analogues, diethylstilbestrol, hydroxyurea. If conservative treatments fail, surgical intervention is required with cavernous shunts. This study reviewed the incidence, pathogenesis and management of sickle cell priapism.

  15. Estrogen and pure antiestrogen fulvestrant (ICI 182 780) augment cell–matrigel adhesion of MCF-7 breast cancer cells through a novel G protein coupled estrogen receptor (GPR30)-to-calpain signaling axis

    Chen, Yan; Li, Zheng; He, Yan; Shang, Dandan; Pan, Jigang; Wang, Hongmei; Chen, Huamei; Zhu, Zhuxia [Department of Physiology/Cancer Research Group, Guiyang Medical University School of Basic Medicine, 9 Beijing Road, Guiyang 550004, Guizhou (China); Wan, Lei [Department of Pharmacology, Guiyang Medical University School of Basic Medicine, 9 Beijing Road, Guiyang 550004, Guizhou (China); Wang, Xudong, E-mail: [Department of Physiology/Cancer Research Group, Guiyang Medical University School of Basic Medicine, 9 Beijing Road, Guiyang 550004, Guizhou (China)


    Fulvestrant (ICI 182 780, ICI) has been used in treating patients with hormone-sensitive breast cancer, yet initial or acquired resistance to endocrine therapies frequently arises and, in particular, cancer recurs as metastasis. We demonstrate here that both 17-beta-estradiol (E2) and ICI enhance cell adhesion to matrigel in MCF-7 breast cancer cells, with increased autolysis of calpain 1 (large subunit) and proteolysis of focal adhesion kinase (FAK), indicating calpain activation. Additionally, either E2 or ICI induced down-regulation of estrogen receptor α without affecting G protein coupled estrogen receptor 30 (GPR30) expression. Interestingly, GPR30 agonist G1 triggered calpain 1 autolysis but not calpain 2, whereas ER agonist diethylstilbestrol caused no apparent calpain autolysis. Furthermore, the actions of E2 and ICI on calpain and cell adhesion were tremendously suppressed by G15, or knockdown of GPR30. E2 and ICI also induced phosphorylation of extracellular regulated protein kinases 1 and 2 (ERK1/2), and suppression of ERK1/2 phosphorylation by U0126 profoundly impeded calpain activation triggered by estrogenic and antiestrogenic stimulations indicating implication of ERK1/2 in the GPR30-mediated action. Lastly, the E2- or ICI-induced cell adhesion was dramatically impaired by calpain-specific inhibitors, ALLN or calpeptin, suggesting requirement of calpain in the GPR30-associated action. These data show that enhanced cell adhesion by E2 and ICI occurs via a novel GPR30-ERK1/2-calpain pathway. Our results indicate that targeting the GPR30 signaling may be a potential strategy to reduce metastasis and improve the efficacy of antiestrogens in treatment of advanced breast cancer. - Highlights: • Estrogen and ICI augment adhesion to matrigel with calpain activation in MCF-7 cells. • GPR30 mediates cell–matrigel adhesion and calpain activation via ERK1/2. • Calpain is required in the cell–matrigel adhesion induced by E2 and ICI.

  16. Levels of 17beta-estradiol receptors expressed in embryonic and adult zebrafish following in vivo treatment of natural or synthetic ligands.

    Gayathri Chandrasekar

    Full Text Available The nuclear receptors encompass a group of regulatory proteins involved in a number of physiological processes. The estrogen receptors (ERs, of which one alpha and one beta form exist in mammals function as transcription factors in response to 17beta-estradiol (E2. In zebrafish there are three gene products of estrogen receptors and they are denoted esr1 (ERalpha, esr2a (ERbeta2 and esr2b (ERbeta1. Total RNA of zebrafish early life stages (<3, 6, 12, 24, 48, 72, 96 and 120 hours post fertilization and of adult fish (liver, intestine, eye, heart, brain, ovary, testis, gill, swim bladder and kidney were isolated following in vivo exposures. Using specific primers for each of the three zebrafish ERs the expression levels were quantified using real time PCR methodology. It was shown that in absence of exposure all three estrogen receptors were expressed in adult fish. The levels of expression of two of these three ER genes, the esr1 and esr2a were altered in organs such as liver, intestine, brain and testis in response to ligand (E2, diethylstilbestrol or 4-nonylphenol. During embryogenesis two of the three receptor genes, esr1 and esr2b were expressed, and in presence of ligand the mRNA levels of these two genes increased. The conclusions are i estrogen receptor genes are expressed during early development ii altered expression of esr genes in response to ligand is dependent on the cellular context; iii the estrogenic ligand 4-nonylphenol, a manufactured compound commonly found in sewage of water treatment plants, acts as an agonist of the estrogen receptor during development and has both agonist and antagonist properties in tissues of adult fish. This knowledge of esr gene function in development and in adult life will help to understand mechanisms of interfering mimicking endocrine chemicals in vivo.

  17. Electrophoretic mobility shift assay on poly(ethylene glycol)-modified glass microchips for the study of estrogen responsive element binding.

    Chuang, Yen-Jun; Huang, Jia-Wei; Makamba, Honest; Tsai, Mei-Ling; Li, Chun-Wei; Chen, Shu-Hui


    The binding of estrogen receptor (ER) to estrogen response element (ERE) is essential for genomic pathways of estrogens and gel-based electrophoretic mobility shift assay (EMSA) is commonly used for analyzing ERE binding. Gel-based EMSA, however, requires the use of hazard radio isotopes and they are slow, labor-intensive and difficult to quantify. Here, we present quantitative affinity assays based on microchip electrophoresis using PEG-modified glass microchannels, which bear neutral surfaces against the adsorption of acidic DNA molecules and basic ER proteins. We first demonstrated the feasibility of the method by measuring binding constants of recombinant ERalpha and ERbeta with a consensus ERE sequence (cERE, 5'-GGTCAGAGTGACC-3') as well as with an ERE-like sequence (ERE 1576, 5'-GACCGGTCAGCGGACTCAC-3'). Changes in mobility as a function of protein-DNA molar ratios were plotted and the dissociation constants were determined based on non-linear curve fitting. The minimum amount of ER proteins required for one assay was around 0.2 ng and the run time for one chip analysis was less than 2 min. We further measured the estrogenic compound-mediated dissociation constants with recombinant ER proteins as well as with the extracted ERbeta from treated and untreated A549 bronchioloalveolar carcinoma cells. Dissociation constants determined by this method agree with the fact that agonist compounds such as 17beta-estradiol (1.70 nM), diethylstilbestrol (0.14 nM), and genistein (0.80 nM) assist ERE binding by decreasing the constants; while antagonist compounds such as testosterone (140.4 nM) and 4-hydroxytamoxifen (10.5 nM) suppress the binding by increasing the dissociation constant.

  18. Alterations in cytochrome P-450 levels in adult rats following neonatal exposure to xenobiotics

    Zangar, R.C. (Oregon State Univ., Corvallis (United States) Pacific Northwest Laboratories, Richland, WA (United States)); Springer, D.L. (Pacific Northwest Laboratories, Richland, WA (United States)); Buhler, D.R. (Oregon State Univ., Corvallis (United States))


    Neonatal exposure to certain xenobiotics has been shown to alter hepatic metabolism in adult rats in a manner that indicates long-term changes in enzyme regulation. Previously, the authors have observed changes in adult testosterone metabolism and in cytochrome P-450 (P-450) mRNA levels in animals neonatally exposed to phenobarbital (PB) or diethylstilbestrol (DES). In order to test for other enzyme alterations, they used Western blot procedures for specific P-450s to analyze hepatic microsomes from adult rats (24 wk old) that had been exposed neonatally to DES, PB, 7,12-dimethylbenz[a]anthracene (DMBA), or pregnenolone 16[alpha]-carbonitrile (PCN). The most striking effects were observed in the DES-treated males: P-4502C6 and an immunologically similar protein were increased 60 and 90%, respectively, relative to control values, but P-4503A2 was decreased by 44%. No changes were observed in the DES-treated males in levels of P-4502E1, P-4502B, or the male-specific P-4502C13. Adult males neonatally treated with PB had 150% increase in levels of anti-P4502B-reactive protein without significant changes in the other enzymes. The DES- and DMBA-treated females had increased levels of the female-specific P-4502C12 of 38 and 48%, respectively, but no other observed alterations. The results confirm that neonatal exposure to DES or PB can cause alterations in adult hepatic cytochrome P-450 levels but show that these chemicals act on different enzymes. Neonatal DMBA resulted in changes in adult females similar to those produced by the synthetic estrogen DES, but did so at about two-thirds lower dose. 37 refs., 5 figs.

  19. Pituitary tumor transforming gene binding factor: a new gene in breast cancer.

    Watkins, Rachel J; Read, Martin L; Smith, Vicki E; Sharma, Neil; Reynolds, Gary M; Buckley, Laura; Doig, Craig; Campbell, Moray J; Lewy, Greg; Eggo, Margaret C; Loubiere, Laurence S; Franklyn, Jayne A; Boelaert, Kristien; McCabe, Christopher J


    Pituitary tumor transforming gene (PTTG) binding factor (PBF; PTTG1IP) is a relatively uncharacterized oncoprotein whose function remains obscure. Because of the presence of putative estrogen response elements (ERE) in its promoter, we assessed PBF regulation by estrogen. PBF mRNA and protein expression were induced by both diethylstilbestrol and 17beta-estradiol in estrogen receptor alpha (ERalpha)-positive MCF-7 cells. Detailed analysis of the PBF promoter showed that the region -399 to -291 relative to the translational start site contains variable repeats of an 18-bp sequence housing a putative ERE half-site (gcccctcGGTCAcgcctc). Sequencing the PBF promoter from 122 normal subjects revealed that subjects may be homozygous or heterozygous for between 1 and 6 repeats of the ERE. Chromatin immunoprecipitation and oligonucleotide pull-down assays revealed ERalpha binding to the PBF promoter. PBF expression was low or absent in normal breast tissue but was highly expressed in breast cancers. Subjects with greater numbers of ERE repeats showed higher PBF mRNA expression, and PBF protein expression positively correlated with ERalpha status. Cell invasion assays revealed that PBF induces invasion through Matrigel, an action that could be abrogated both by siRNA treatment and specific mutation. Furthermore, PBF is a secreted protein, and loss of secretion prevents PBF inducing cell invasion. Given that PBF is a potent transforming gene, we propose that estrogen treatment in postmenopausal women may upregulate PBF expression, leading to PBF secretion and increased cell invasion. Furthermore, the number of ERE half-sites in the PBF promoter may significantly alter the response to estrogen treatment in individual subjects.

  20. Mouse hypospadias: A critical examination and definition.

    Sinclair, Adriane Watkins; Cao, Mei; Shen, Joel; Cooke, Paul; Risbridger, Gail; Baskin, Laurence; Cunha, Gerald R


    Hypospadias is a common malformation whose etiology is based upon perturbation of normal penile development. The mouse has been previously used as a model of hypospadias, despite an unacceptably wide range of definitions for this malformation. The current paper presents objective criteria and a definition of mouse hypospadias. Accordingly, diethylstilbestrol (DES) induced penile malformations were examined at 60 days postnatal (P60) in mice treated with DES over the age range of 12 days embryonic to 20 days postnatal (E12-P20). DES-induced hypospadias involves malformation of the urethral meatus, which is most severe in DES E12-P10, DES P0-P10 and DES P5-P15 groups, and less so or absent in the other treatment groups. A frenulum-like ventral tether between the penis and the prepuce was seen in the most severely affected DES-treated mice. Internal penile morphology was also altered in the DES E12-P10, DES P0-P10 and DES P5-P15 groups (with little effect in the other DES treatment groups). Thus, adverse effects of DES are a function of the period of DES treatment and most severe in the P0-P10 period. In "estrogen mutant mice" (NERKI, βERKO, αERKO and AROM+) hypospadias was only seen in AROM+ male mice having genetically-engineered elevation is serum estrogen. Significantly, mouse hypospadias was only seen distally at and near the urethral meatus where epithelial fusion events are known to take place and never in the penile midshaft, where urethral formation occurs via an entirely different morphogenetic process.

  1. Effect of 1,25-Dihydroxyvitamin D3 on Regulatory T Cells in Ovariectomized Mice

    LIU Jun Chen; ZHOU Chen Hui; ZHANG Xue; CHEN Yan; XU Bi Lian; CUI Liao; XU Dao Hua


    ObjectiveTo investigate thecorrelation betweenregulatory T(Treg) cellsand postmenopausal osteoporosis andthe antiosteoporotic effect of 1,25-dihydroxyvitamin D3[1,25(OH)2D3] in relation to Treg cells. MethodsFifty femaleBALB/c mice were randomly divided into five groups: the basal control (BAS), Sham,ovariectomy (OVX),OVX+diethylstilbestrol (OVX+DES), andOVX+1,25(OH)2D3.Tibias were harvested andprocessed with decalcification for quantitative bone histomorphometry.Femurs were stained by immunohistochemistry to detectFoxp3 protein expression.Spleens wereused to detect Treg andFoxp3 gene expressionby flow cytometry andquantitative RT-PCR, respectively. ResultsIn comparison withthe Sham group,a significant decrease was found inthe OVX group in such indices as trabecular bone volume/total tissue area (BV/TV), trabecularnumber (Tb.N) and trabecular thickness (Tb.Th).1,25(OH)2D3andDESpartlyprevented the decrease in BV/TV, Tb.N, Tb.Th inOVX mice.Treg cell number, Foxp3 mRNA expression in spleen andFoxp3 protein expression in femur significantly decreasedinthe OVX-treated group compared withthose in thesham group.1,25(OH)2D3 andDES significantlyincreased Treg cell number andFoxp3 expression.Treg cellsand Foxp3 gene expression were related to bonehistomorphometricparameters. ConclusionThedecrease inTreg cell numbersis relevant to the postmenopausal osteoporosis. The antiosteoporosis of1,25(OH)2D3 is related to regulatory T cells.

  2. Avian WNT4 in the female reproductive tracts: potential role of oviduct development and ovarian carcinogenesis.

    Chul-Hong Lim

    Full Text Available The wingless-type MMTV integration site family of proteins (WNTs is highly conserved secreted lipid-modified signaling molecules that play a variety of pivotal roles in developmental events such as embryogenesis, tissue homeostasis and cell polarity. Although, of these proteins, WNT4 is known to be involved in genital development in fetuses of mammalian species, its role is unknown in avian species. Therefore, in this study, we investigated expression profiles, as well as hormonal and post-transcriptional regulation of WNT4 expression in the reproductive tract of female chickens. Results of this study demonstrated that WNT4 is most abundant in the stromal and luminal epithelial cells of the isthmus and shell gland of the oviduct, respectively. WNT4 is also most abundant in the glandular epithelium of the shell gland of the oviduct of laying hens at 3 h post-ovulation during the laying cycle. In addition, treatment of young chicks with diethylstilbestrol (DES, a synthetic estrogen agonist stimulated WNT4 only in the glandular epithelial cells of the isthmus and shell gland of the oviduct. Moreover, results of our study demonstrated that miR-1786 influences WNT4 expression via specific binding sites in its 3'-UTR. On the other hand, our results also indicate that WNT4 is expressed predominantly in the glandular epithelium of cancerous ovaries, but not in normal ovaries of hens. Collectively, these results indicate cell-specific expression of WNT4 in the reproductive tract of chickens and that it likely has crucial roles in development and function of oviduct as well as initiation of ovarian carcinogenesis in laying hens.

  3. Comparison of Short-Term Estrogenicity Tests for Identification of Hormone-Disrupting Chemicals

    Andersen, Helle Raun; Andersson, Anna-Maria; Arnold, Steven F.; Autrup, Herman; Barfoed, Marianne; Beresford, Nicola A.; Bjerregaard, Poul; Christiansen, Lisette B.; Gissel, Birgitte; Hummel, René; Jørgensen, Eva Bonefeld; Korsgaard, Bodil; Le Guevel, Remy; Leffers, Henrik; McLachlan, John; Møller, Anette; Bo Nielsen, Jesper; Olea, Nicolas; Oles-Karasko, Anita; Pakdel, Farzad; Pedersen, Knud L.; Perez, Pilar; Skakkebœk, Niels Erik; Sonnenschein, Carlos; Soto, Ana M.; Sumpter, John P.; Thorpe, Susan M.; Grandjean, Philippe


    The aim of this study was to compare results obtained by eight different short-term assays of estrogenlike actions of chemicals conducted in 10 different laboratories in five countries. Twenty chemicals were selected to represent direct-acting estrogens, compounds with estrogenic metabolites, estrogenic antagonists, and a known cytotoxic agent. Also included in the test panel were 17β-estradiol as a positive control and ethanol as solvent control. The test compounds were coded before distribution. Test methods included direct binding to the estrogen receptor (ER), proliferation of MCF-7 cells, transient reporter gene expression in MCF-7 cells, reporter gene expression in yeast strains stably transfected with the human ER and an estrogen-responsive reporter gene, and vitellogenin production in juvenile rainbow trout. 17β-Estradiol, 17α-ethynyl estradiol, and diethylstilbestrol induced a strong estrogenic response in all test systems. Colchicine caused cytotoxicity only. Bisphenol A induced an estrogenic response in all assays. The results obtained for the remaining test compounds—tamoxifen, ICI 182.780, testosterone, bisphenol A dimethacrylate, 4-n-octylphenol, 4-n-nonylphenol, nonylphenol dodecylethoxylate, butylbenzylphthalate, dibutylphthalate, methoxychlor, o,p′-DDT, p,p′-DDE, endosulfan, chlomequat chloride, and ethanol—varied among the assays. The results demonstrate that careful standardization is necessary to obtain a reasonable degree of reproducibility. Also, similar methods vary in their sensitivity to estrogenic compounds. Thus, short-term tests are useful for screening purposes, but the methods must be further validated by additional interlaboratory and interassay comparisons to document the reliability of the methods. ImagesFigure 2Figure 5Figure 6Figure 7 PMID:10229711

  4. Effect of galactosamine-induced hepatic UDP-glucuronic acid depletion on acetaminophen elimination in rats. Dispositional differences between hepatically and extrahepatically formed glucuronides of acetaminophen and other chemicals.

    Gregus, Z; Madhu, C; Goon, D; Klaassen, C D


    Galactosamine (GAL) markedly depletes hepatic UDP-glucuronic acid (UDP-GA) whereas extrahepatic UDP-GA is minimally affected. This suggests that GAL predominantly inhibits hepatic glucuronidation. Therefore, the effect of GAL-induced hepatic UDP-GA depletion was examined in bile duct-cannulated rats to determine the role of hepatic glucuronidation in the disposition of acetaminophen (AA). GAL markedly altered the fate of AA-glucuronide but had little or no effect upon other AA metabolites. GAL decreased the biliary excretion of AA-glucuronide up to 92%, whereas reductions in blood levels and urinary excretion of AA-glucuronide did not exceed 50%. This suggests that AA-glucuronide excreted in bile is predominantly of hepatic origin whereas AA-glucuronide found in blood and urine is derived from both hepatic and extrahepatic tissues. Data in the present and previous studies [Gregus, Watkins, Thompson, Klaassen: J. Pharmacol. Exp. Ther. 225, 256, (1983)] indicate that GAL greatly reduced the biliary excretion of AA- and valproic acid-glucuronide whereas the biliary excretion of the glucuronides of phenolphthalein, iopanoic acid, bilirubin, and diethylstilbestrol was only partially decreased. This difference appears to be largely due to differential contributions by the liver and extrahepatic tissues in the glucuronidation of various compounds as well as the availability of glucuronides formed in extrahepatic tissues for biliary excretion. Specifically, the extrahepatically formed glucuronide conjugates of AA and valproic acid are not readily available for biliary excretion whereas the glucuronides of the other compounds are readily excreted into bile.(ABSTRACT TRUNCATED AT 250 WORDS)

  5. Assessment of source water contamination by estrogenic disrupting compounds in China

    Weiwei Jiang; Ye Yan; Mei Ma; Donghong Wang; Qian Luo; Zijian Wang; Senthil Kumaran Satyanarayanan


    Detection of estrogenic disrupting compounds (EDCs) in drinking waters around China has led to rising concerns about health risks associated with these compounds.There is,however,a paucity of studies on the occurrence and identification of the main compounds responsible for this pollution in the source waters.To fill this void,we screened estrogenic activities of 23 source water samples from six main river systems in China,using a recombinant two-hybrid yeast assay.All sample extracts induced significant estrogenic activity,with E2 equivalents (EEQ) of raw water ranging from 0.08 to 2.40 ng/L.Additionally,16 samples were selected for chemical analysis by gas chromatography-mass spectrometry.The EDCs of most concern,including estrone (El),17β-estradiol (E2),17α-ethinylestradiol (EE2),estriol (E3),diethylstilbestrol (DES),estradiol valerate (EV),4-t-octylphenol (4-t-OP),4-nonylphenols (4-NP) and bisphenol A (BPA),were determined at concentrations of up to 2.98,1.07,2.67,4.37,2.52,1.96,89.52,280.19 and 710.65 ng/L,respectively.Causality analysis,involving comparison of EEQ values from yeast assay and chemical analysis identified E2,EE2 and 4-NP as the main responsible compounds,accounting for the whole estrogenic activities (39.74% to 96.68%).The proposed approach using both chemical analysis and yeast assay could be used for the identification and evaluation of EDCs in source waters of China.

  6. Xenoestrogens at picomolar to nanomolar concentrations trigger membrane estrogen receptor-alpha-mediated Ca2+ fluxes and prolactin release in GH3/B6 pituitary tumor cells.

    Wozniak, Ann L; Bulayeva, Nataliya N; Watson, Cheryl S


    Xenoestrogens (XEs) are widespread in our environment and are known to have deleterious effects in animal (and perhaps human) populations. Acting as inappropriate estrogens, XEs are thought to interfere with endogenous estrogens such as estradiol (E2) to disrupt normal estrogenic signaling. We investigated the effects of E2 versus several XEs representing organochlorine pesticides (dieldrin, endosulfan, o',p'-dichlorodiphenylethylene), plastics manufacturing by-products/detergents (nonylphenol, bisphenol A), a phytoestrogen (coumestrol), and a synthetic estrogen (diethylstilbestrol) on the pituitary tumor cell subline GH3/B6/F10, previously selected for expression of high levels of membrane estrogen receptor-alpha. Picomolar to nanomolar concentrations of both E2 and XEs caused intracellular Ca2+ changes within 30 sec of administration. Each XE produced a unique temporal pattern of Ca2+ elevation. Removing Ca2+ from the extracellular solution abolished both spontaneous and XE-induced intracellular Ca2+ changes, as did 10 microM nifedipine. This suggests that XEs mediate their actions via voltage-dependent L-type Ca2+ channels in the plasma membrane. None of the Ca2+ fluxes came from intracellular Ca2+ stores. E2 and each XE also caused unique time- and concentration-dependent patterns of prolactin (PRL) secretion that were largely complete within 3 min of administration. PRL secretion was also blocked by nifedipine, demonstrating a correlation between Ca2+ influx and PRL secretion. These data indicate that at very low concentrations, XEs mediate membrane-initiated intracellular CCa2+ increases resulting in PRL secretion via a mechanism similar to that for E2, but with distinct patterns and potencies that could explain their abilities to disrupt endocrine functions.

  7. In vivo induction of oocyte maturation and ovulation in zebrafish.

    Toshinobu Tokumoto

    Full Text Available The maturation of fish oocytes is a well-characterized system induced by progestins via non-genomic actions. In a previous study, we demonstrated that diethylstilbestrol (DES, a non-steroidal estrogen, induces fish oocyte maturation via the membrane progestin receptor (mPR. Here, we attempted to evaluate the effect of DES as an environmental endocrine disrupting chemical (EDC upon fish oocyte maturation using live zebrafish. DES triggered oocyte maturation within several hours in vivo when administrated directly into the surrounding water. The natural teleost maturation-inducing hormone, 17alpha, 20beta-dihydroxy-4-pregnen-3-one (17,20beta-DHP also induced oocyte maturation in vivo. Steroids such as testosterone, progesterone or 17alpha-hydroxyprogesterone were also effective in vivo. Further studies indicated that externally applied 17,20beta-DHP even induced ovulation. In contrast to 17,20beta -DHP, DES induced maturation but not ovulation. Theoretically this assay system provides a means to distinguish pathways involved in the induction of ovulation, which are known to be induced by genomic actions from the pathway normally involved in the induction of oocyte maturation, a typical non-genomic action-dependent pathway. In summary, we have demonstrated the effect of EDCs on fish oocyte maturation in vivo. To address the effects, we have explored a conceptually new approach to distinguish between the genomic and non-genomic actions induced by steroids. The assay can be applied to screens of progestin-like effects upon oocyte maturation and ovulation for small molecules of pharmacological agents or EDCs.

  8. Rapid signaling actions of environmental estrogens in developing granule cell neurons are mediated by estrogen receptor ß.

    Le, Hoa H; Belcher, Scott M


    Estrogenic endocrine disrupting chemicals (EDCs) constitute a diverse group of man-made chemicals and natural compounds derived from plants and microbial metabolism. Estrogen-like actions are mediated via the nuclear hormone receptor activity of estrogen receptor (ER)α and ERβ and rapid regulation of intracellular signaling cascades. Previous study defined cerebellar granule cell neurons as estrogen responsive and that granule cell precursor viability was developmentally sensitive to estrogens. In this study experiments using Western blot analysis and pharmacological approaches have characterized the receptor and signaling modes of action of selective and nonselective estrogen ligands in developing cerebellar granule cells. Estrogen treatments were found to briefly increase ERK1/2-phosphorylation and then cause prolonged depression of ERK1/2 activity. The sensitivity of granule cell precursors to estrogen-induced cell death was found to require the integrated activation of membrane and intracellular ER signaling pathways. The sensitivity of granule cells to selective and nonselective ER agonists and a variety of estrogenic and nonestrogenic EDCs was also examined. The ERβ selective agonist DPN, but not the ERα selective agonist 4,4',4'-(4-propyl-[1H]-pyrazole-1,3,5-triyl) trisphenol or other ERα-specific ligands, stimulated cell death. Only EDCs with selective or nonselective ERβ activities like daidzein, equol, diethylstilbestrol, and bisphenol A were observed to induce E2-like neurotoxicity supporting the conclusion that estrogen sensitivity in granule cells is mediated via ERβ. The presented results also demonstrate the utility of estrogen sensitive developing granule cells as an in vitro assay for elucidating rapid estrogen-signaling mechanisms and to detect EDCs that act at ERβ to rapidly regulate intracellular signaling.

  9. Comparison of an array of in vitro assays for the assessment of the estrogenic potential of natural and synthetic estrogens, phytoestrogens and xenoestrogens.

    Gutendorf, B; Westendorf, J


    Many chemicals in surface waters and sediments have recently been discovered to have estrogenic/antiestrogenic activity. Among these compounds, known as 'endocrine disrupters', are natural and synthetic hormones, phytoestrogenes and a variety of industrial chemicals, such as certain detergents and pesticides. These substances are supposed to affect the development and reproduction in wildlife and humans and may also be involved in the induction of cancer. In order to assess the estrogenic/antiestrogenic potential of pure compounds and complex environmental samples we compared an array of in vitro test systems, (i) two luciferase reporter gene assays using transgenic human MVLN cells (derived from MCF-7 cells) and HGELN cells (derived from HeLa cells); (ii) a competitive binding assay with recombinant human estrogen receptors (ER) alpha and beta; and (iii) a proliferation assay with MCF7-cells (E-Screen). The sensitivity of the assays for 17-beta-estradiol decreased in the order: MVLN-cells=E-Screen>HGELN-cells>binding to ER-alpha>binding to ER-beta. A good correlation was obtained between the estrogenic potencies of 11 compounds (17-beta-estradiol (E(2)), estrone (E(1)), estriol (E(3)), ethinylestradiol (EE(2)), diethylstilbestrol (DES), coumestrol, beta-sitosterol, genistein, 4-nonylphenol, 4-octylphenol, bisphenol A) in the three tissue culture assays. The relative potencies of the compounds obtained by the cell free binding assays were one to two orders of magnitude higher compared with the cell culture assays. The phytoestrogens showed a preference to bind to ER-beta, but only genistein showed a much lower activity in the E-Screen (growth induction in breast cancer cells) compared with the luciferase induction in MVLN and HGELN-cells.

  10. Autocrine role of estrogens in the augmentation of luteinizing hormone receptor formation in cultured rat granulosa cells.

    Kessel, B; Liu, Y X; Jia, X C; Hsueh, A J


    The effects of estrogens on gonadotropin-stimulated luteinizing hormone (LH) receptor formation were examined in primary cultures of rat granulosa cells. Granulosa cells were cultured for 3 days with increasing concentrations of follicle-stimulating hormone (FSH) in the presence or absence of native and synthetic estrogens. Follicle-stimulating hormone stimulated LH receptor formation in a dose-dependent fashion, and estrogens enhanced the FSH-stimulated LH receptor content by decreasing the apparent ED50 of FSH. At 6.25 ng/ml FSH, the enhancement in LH receptor was estrogen dose dependent, with an ED50 value of about 3 X 10(-9) M for 17 beta-estradiol. The increased LH receptor content seen in cells treated with FSH and estrogen was correlated with increased cAMP production by these cells in response to LH stimulation. Time course studies revealed enhancement of FSH-stimulated LH receptor induction at 48 and 72 h of culture. Granulosa cells were also cultured with FSH for 2 days to induce functional LH receptors, then further cultured for 3 days with LH in the presence or absence of estrogens. At 30 ng/ml LH, increasing concentrations of estrogens maintained LH receptor content in a dose-dependent fashion, with their relative estrogenic potencies in keeping with reported binding affinities to estrogen receptors. An autocrine role of estrogens on LH receptor formation was further tested in granulosa cells treated with FSH and an aromatase substrate (androstenedione) to increase estrogen biosynthesis. Cotreatment with semipurified estrogen antibodies partially blocked the FSH stimulation of LH receptors, whereas nonimmune serum was ineffective. Also, inclusion of diethylstilbestrol prevented the inhibitory effect of the estrogen antibodies. Thus, local estrogens in ovarian follicles may play an autocrine role in granulosa cells to enhance LH receptor formation and to increase granulosa cell responsiveness to the LH surge, with subsequent ovulation and adequate

  11. Effects of compound piracetam in preventing osteoporosis in ovariectomized rats%复方脑复康预防大鼠去卵巢后骨质疏松的研究

    黄健萍; 林思恩; 吴玲芝; 崔燎; 吴铁


    目的:探讨复方脑复康防治去卵巢后骨质疏松的效果.方法:4月龄Sprague-dawley (SD)雌性大鼠38只随机分为5组:基础组,6只,于实验前取材;假手术组,8只,给予生理盐水灌胃;去卵巢组,8只,去卵巢后给予生理盐水灌胃;已烯雌酚组,8只,去卵巢后给予已烯雌酚30 μg/(kg·d)灌胃;复方脑复康组,8只,去卵巢后给予吡拉西坦432mg/(kg·d)和司坦唑醇0.54mg/(kg·d)灌胃.连续灌胃90d后,处死大鼠并取左胫骨包埋、切片、染色后作静态和动态骨组织形态计量学测定.结果:与假手术组相比,去卵巢组骨小梁面积百分数(%Tb.Ar)、骨小梁厚度(Tb.Th)、骨小梁数目(Tb.N)和标记周长百分数(%L.Pm)均显著降低(P<0.01),骨小梁分离度(Tb.Sp)增加(均P<0.01);每毫米破骨细胞数目(Oc.N)和破骨细胞周长百分数(%Oc.Pm)均显著增加(P<0.01);骨形成率(BFR/BV)增加(P<0.05).与去卵巢组相比,已烯雌酚及复方脑复康都可使%Tb.Ar显著增加(均P<0.01),Tb.N显著增加(均P<0.01),Tb.Th增加(P>0.05,P<0.05);并使Tb.Sp显著降低(均P<0.01);使Oc.N和%Oc.Pm显著减少(均P<0.01).与已烯雌酚组相比,复方脑复康可以使胫骨骨小梁Tb.Th和%L.Pm增加(均P<0.05).结论:复方脑复康可以预防大鼠去卵巢后骨质疏松,其作用效应与雌激素相当,作用机制可能与复方脑复康促进骨形成,抑制骨吸收有关.%Objective:To examine the effects of compound piracetam(P-S) in preventing cancellous bone loss in ovariectomized (ovx) rats. Methods:Thirty eight female Sprague-dawley rats of four months old were randomly divided into five groups: the basal control (BAS,n=6),sham+saline(SHAM,n=8),ovx+saline(OVX,n=8),ovx +diethylstilbestrol 30 μg/(kg·d) (0VX+DES,n=8),and ovx+ Piracetam 432 mg/(kg·d)+ stanozolol 0.54 mg/(kg·d)(OVX+P-S,n=8). Rats were treated by oral gavage for 90 days.The basal control were sacrificed at day 0; other groups were killed

  12. 盐酸米诺环素合并己烯雌酚治疗痤疮疗效观察



    The acne is a chronic skin disease, with acne, Qiu diagnosis, pustules, nodules, cysts and scars characteristics. The cause of the excessive secretion of androgens as the main role, minocycline as broad-spectrum antibiotics, has a strong inhibitory effect on acne bacilus, and can achieve effective antibiotic concentration in sebaceous gland. For the synthetic estrogen diethylstilbestrol, counteract the androgen action, have tender skin, relieve skin hyperkeratosis effect. Different dimension A acid is a vitamin A derivative, topical invalid, oral administration can inhibit the sebaceous gland function, and can make the skin P.acnes significantly reduced. But the side effects of this product is larger, can cause dry skin, itching, desquamation, temporary hair loss, there are a few joint and muscle pain, fatigue anorexia, this product and the distortion effect. Diethylstilbestrol can be absorbed through the skin, mucosa and muscle, fat soluble. Therefore, the application of diethylstilbestrol external, not oral, drugs directly to the skin, directly on the target cel, not only of good absorption, but also reduces the oral administration of anti androgen effect on human body, reduce the impact on the endocrine. This product is also reduce hyperkeratotic skin rejuvenation, improve the diagonalization and catheter hair folicle and sebaceous gland abnormalities, reduce sebum excretion barriers, reduction of Propionibacterium reproductive survival environment, thereby reducing the generation of mediators of inflammation and inflammation. Cheap and control the comparison, obvious advantages,, less side effect, is worth promoting.%痤疮是一种慢性皮肤病,多以粉刺,丘诊,脓疱,结节,囊肿及瘢痕为特征。其病因以雄激素分泌过多为主要作用,米诺环素为广谱抗生素,对痤疮杆菌有较强的抑制作用,并能在皮脂腺中达到有效抗菌浓度。己烯雌酚为人工合成雌激素,有抵消雄激素作用,有嫩

  13. Breast cancer epidemiology.

    Kelsey, J L; Berkowitz, G S


    The various risk factors for breast cancer have been recognized for many years. A table lists these established breast cancer risk factors together with the approximate magnitude of the increase in risk associated with them. Breast cancer incidence rates increase with age throughout the life span in Western countries, although the rate of increase is greater up to age 50 years than after 50 years. Breast cancer is more common among women in upper rather than lower social classes, among women who never have been married, among women living in urban areas, among women living in the northern US than in the southern US, and among whites than blacks, at least among those over age 50. Women in North American and Northern European countries have the highest risk for breast cancer, women in Southern European and Latin American countries are at intermediate risk, and women in Africa and Asian countries have the lowest risk. Yet, rapid rates of increase in incident rates have been noted in recent years in many Asian, Central European, and some South American countries. The later the age at which a woman has her 1st full-term pregnancy, the higher her risk for breast cancer; the earlier the age at menarche and the later the age at menopause the higher the risk; and among women who have a premenopausal oophorectomy, the earlier the age at which this occurs the lower the risk. Among postmenopausal women, obesity is associated with an increase in risk. Lactation is negatively associated with subsequent breast cancer risk. Some current research is considering potential risk factors that have not been well studied in the past, including alcohol consumption, cigarette smoking, caffeine consumption, exposure to diethylstilbestrol (DES), emotional stress, exposure to electric power, and lack of physical activity. Other areas of current research reviewed here include radiation, mammographic parenchymal patterns, a high-fat diet, use of oral contraceptives (OCs), use of estrogen

  14. Clear cell carcinoma of the uterine cervix: clinical characteristics and feasibility of fertility-preserving treatment

    Jiang X


    Full Text Available Xiang Jiang, Ying Jin, Yan Li, Hui-Fang Huang, Ming Wu, Keng Shen, Ling-Ya Pan Department of Obstetrics and Gynecology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, People's Republic of China Abstract: The objective of this retrospective study was to analyze the clinical characteristics and prognosis of clear cell adenocarcinoma (CCA in the post-diethylstilbestrol (DES era and to evaluate the feasibility of fertility-preserving treatment. The records of 32 patients with CCAs who were treated at Peking Union Medical College Hospital from August 1986 to June 2012 were retrospectively reviewed. Three of the patients had undergone fertility-preserving treatment. The incidence of CCA among cervical adenocarcinomas was 15.2%. The median age was 38 years: 11 patients (34.4% were diagnosed before 30 years of age and two (6.3% after 70 years of age. Ten patients (31.2% were nulliparous. No patient had been exposed to DES. Twenty-nine patients (90.6% presented with obvious symptoms, and the cervix appeared abnormal in 26 patients (81.3%. Cervical Papanicolaou (Pap tests were abnormal in all four patients in whom they were performed (three had high-grade squamous intraepithelial lesions and one had atypical squamous cells of undetermined significance. The distribution by stage was 56.3% stage I, 34.4% stage II, 6.3% stage III, and 3.1% stage IV. Treatments mainly included surgery for patients with stage I to IIA CCA and radiochemotherapy for patients with advanced CCA. The overall 5-year progression-free survival was 72.2%. Patients with stage I to IIA CCA had better 5-year progression-free survival than did patients with stage IIB to IV CCA (81.5% versus 40.0%, P=0.003. The three patients who had undergone fertility-preserving treatment had no recurrences. CCA may also affect adolescents and children without prior DES exposure, who are often misdiagnosed as having functional uterine

  15. Influence of exercise on bone remodeling-related hormones and cytokines in ovariectomized rats: a model of postmenopausal osteoporosis.

    Li, Lihui; Chen, Xi; Lv, Shuang; Dong, Miaomiao; Zhang, Li; Tu, Jiaheng; Yang, Jie; Zhang, Lingli; Song, Yinan; Xu, Leiting; Zou, Jun


    This study aims to explore the effects of exercise on postmenopausal osteoporosis and the mechanisms by which exercise affects bone remodeling. Sixty-three Wistar female rats were randomly divided into five groups: (1) control group, (2) sham-operated group, (3) OVX (Ovariectomy) group, (4) DES-OVX (Diethylstilbestrol-OVX) group, and (5) Ex-OVX (Exercise-OVX) group. The rat osteoporosis model was established through ovariectomy. The Ex-OVX rats were made to run 251.2 meters every day, 6 d/wk for 3 months in a running wheel. Trabecular bone volume (TBV%), total resorption surface (TRS%), trabecular formation surface (TFS%), mineralization rate (MAR), bone cortex mineralization rate (mAR), and osteoid seam width (OSW) were determined by bone histomorphometry. The mRNA and protein levels of interleukin-1β (IL-1β2), interleukin-6 (IL-6), and cyclooxygenase-2 (Cox-2) were determined by in situ hybridization and immunohistochemistry, respectively. Serum levels of estrogen estradiol (E2), calcitonin (CT), osteocalcin (BGP), and parathyroid hormone (PTH) were determined by ELISA assays. The investigation revealed that compared to the control and the sham-operated groups, the OVX group showed significantly lower levels of TBV%, E2, and CT, but much higher levels of TRS%, TFS%, MAR, OSW, BGP, and PTH. The Ex-OVX group showed increased TBV% and serum levels of E2 and CT compared to the OVX group. Ovariectomy also led to a significant increase in IL-1β mRNA and protein levels in the bone marrow and IL-6 and Cox-2 protein levels in tibias. In addition, the Ex-OVX group showed lower levels of IL-1 mRNA and protein, IL-6 mRNA, and Cox-2 mRNA and protein than those in the OVX group. The upshot of the study suggests that exercise can significantly increase bone mass in postmenopausal osteoporosis rat models by inhibiting bone resorption and increasing bone formation, especially in trabecular bones.

  16. High estrogen concentrations in receiving river discharge from a concentrated livestock feedlot.

    Chen, Te-San; Chen, Ting-Chien; Yeh, Kuei-Jyum C; Chao, How-Ran; Liaw, Ean-Tun; Hsieh, Chi-Ying; Chen, Kuan-Chung; Hsieh, Lien-Te; Yeh, Yi-Lung


    Environmental estrogenic chemicals interrupt endocrine systems and generate reproductive abnormalities in wildlife, especially natural and synthetic estrogenic steroid hormones such as 17beta-estradiol (E2), estrone (E1), estriol (E3), 17alpha-ethynylestradiol (EE2), and diethylstilbestrol (DES). Concentrated animal feedlot operations (CAFOs) are of particular concern since large amounts of naturally excreted estrogens are discharged into aquatic environments. This study investigated E2, E1, E3, EE2, and DES with high performance liquid chromatography/tandem mass (HPLC-MS/MS) analyses along Wulo Creek in southern Taiwan, near a concentrated livestock feedlot containing 1,030,000 broiler chickens, 934,000 laying hens, 85,000 pigs, and 1500 cattle. Sampling was performed from December 2008 to May 2009, in which 54 samples were collected. Experimental results indicate that concentrations of EE2 were lower than the limit of detection (LOD), and concentrations of DES were only detected twice. Concentrations ranged from 7.4 to 1267 ng/L for E1, from not detected (ND) to 313.6 ng/L for E2, and from ND to 210 ng/L for E3. E1 had the highest average mass fraction (72.2 + or - 3.6%), which was significantly higher than E3 (16.2 + or - 1.7%) and E2 (11.5 + or - 2.6%). Additionally, the mean E2 equivalent quotient (EEQ) ranged from 17.3 to 137.9 ng-E2/L. Despite having a markedly lower concentration than E1, E2 more significantly contributed (52.4 + or - 6.0%) EEQ than E1 (19.7 + or - 3.5%). Moreover, the concentrations of E2, E1, and E3 upstream were significantly higher than concentrations downstream, suggesting a high attenuation effect and fast degradation in the study water. Most concentrations in winter season were higher than those of spring season due to the low dilution effect and low microbial activity in the winter season. Based on the results of this study, we recommend further treatment of the wastewater discharge from the feedlot.

  17. 武汉地区水中环境内分泌干扰物的污染情况及其对人体的影响%Analysis on pollution of environmental endocrine disruptors in water in Wuhan area and its influence on human body

    叶超; 刘燕群; 游浩


    Wuhan area has rich water resource and vast water area, while water is closely related to people's daily lives. So the water pollution is very important to life quality of residents in Wuhan, and the scientists pay more attention to the water environment in Wuhan area. In recent years, the pollution of environmental endocrine disruptors in Wuhan area has become more serious, and the environmental endocrine disruptors more and more attracted people's attention. The environmental endocrine disruptors, which include estrone (El) , 17 α-acetylene estrone (EE2) , estradiol (E2) , diethylstilbestrol (DES) , nonylphenol (NP) , octyl phenol ( OP) , bisphenol A ( BPA) , Di-n-Butyl phthalate( DBP) and di-(2-ethylhcxyl) phthalate ( DEHP) , widely distribute in Wuhan area, and have different effect on human body. The paper makes a brief analysis on pollution of environmental endocrine disruptors in Wuhan area and its influence on human body.%武汉水资源总量丰富,水域面在,而水又与人们的日常生活息息相关,因而武汉的水环境污染情况对武汉人的生活状况十分重要,武汉地区的水环境也一直是科学家们的重要关注对象.近年来武汉地区环境内分泌干扰物污染情况比较严重,环境内分泌干扰物越来越引起人们的重视.环境内分泌干扰物有很多种,如雌酮(EI)、17α-乙炔雌酮(EE2)、雌二醇(E2)、已烯雌酚(DES)、壬基本分、辛基酚(OP)、双酚A(BPA)、酞酸二正丁酯(DBP)、酞酸二(2-乙基己基)酯(DEHP)等,它们在武汉的分布情况各有不同,对人体的影响也不一样,在这里就武汉地区环境内分泌干扰物的污染情况及其对人体的影响进行简要分析.

  18. Comparison of the expression profiles induced by genotoxic and nongenotoxic carcinogens in rat liver

    Ellinger-Ziegelbauer, Heidrun [Bayer Healthcare AG, Department of Molecular and Genetic Toxicology, Aprather Weg 18a, 42096 Wuppertal (Germany)]. E-mail:; Stuart, Barry [Bayer Crop Science, Department of Toxicology, Stilwell, KS (United States); Wahle, Brad [Bayer Crop Science, Department of Toxicology, Stilwell, KS (United States); Bomann, Werner [Bayer Crop Science, Department of Toxicology, Stilwell, KS (United States); Ahr, Hans Juergen [Bayer Healthcare AG, Department of Molecular and Genetic Toxicology, Aprather Weg 18a, 42096 Wuppertal (Germany)


    Application of recently developed gene expression techniques using microarrays in toxicological studies (toxicogenomics) facilitate the interpretation of a toxic compound's mode of action and may also allow the prediction of selected toxic effects based on gene expression changes. In order to test this hypothesis, we investigated whether carcinogens at doses known to induce liver tumors in the 2-year rat bioassay deregulate characteristic sets of genes in a short term in vivo study and whether these deregulated genes represent defined biological pathways. Male Wistar rats were dosed with the four nongenotoxic hepatocarcinogens methapyrilene (MPy, 60 mg/kg/day), diethylstilbestrol (DES, 10 mg/kg/day), Wy-14643 (Wy, 60 mg/kg/day), and piperonylbutoxide (PBO, 1200 mg/kg/day). After 1, 3, 7, and 14 days, the livers were taken for histopathological evaluation and for analysis of the gene expression profiles on Affymetrix RG{sub U}34A arrays. The expression profile of the four nongenotoxic carcinogens were compared to the profiles of the four genotoxic carcinogens 2-nitrofluorene (2-NF), dimethylnitrosamine (DMN), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), and aflatoxin B1 (AB1) from a similar study reported previously. By using statistical and clustering tools characteristically deregulated genes were extracted and functionally classified. Distinct cellular pathways were affected by the nongenotoxic carcinogens compared to the genotoxic carcinogens which at least partly correlated with the two-stage model of carcinogenesis. Characteristic to genotoxic carcinogens were a DNA damage response and the activation of proliferative and survival signaling. Nongenotoxic carcinogens showed responses to oxidative DNA or protein damage, as well as cell cycle progression and signs of regeneration. Many of the gene alterations found with the nongenotoxic carcinogens imply compound-specific mechanisms. Although neither a single gene nor a single pathway will be

  19. Targeted analysis with benchtop quadrupole–orbitrap hybrid mass spectrometer: Application to determination of synthetic hormones in animal urine

    Kumar, Praveen [Departament de Química Analítica, Universitat de Barcelona, Barcelona (Spain); Rúbies, Antoni; Centrich, Francesc [Laboratori Agència Salut Pública de Barcelona, Barcelona (Spain); CIBER of Epidemiology and Public Health (CIBERESP), Madrid (Spain); Granados, Mercè [Departament de Química Analítica, Universitat de Barcelona, Barcelona (Spain); Cortés-Francisco, Nuria; Caixach, Josep [Mass Spectrometry Laboratory-Organic Pollutants, IDAEA-CSIC, Barcelona (Spain); Companyó, Ramon, E-mail: [Departament de Química Analítica, Universitat de Barcelona, Barcelona (Spain)


    Graphical abstract: -- Highlights: •The quadrupole in Q Exactive acts as a powerful filter to reduce ion suppression. •Reducing mass range using quadrupole in targeted modes increases the S/N ratio. •Targeted SIM data dependent scan modes are the most suitable for residue analysis. •A HRMS confirmatory method for synthetic hormones in urine has been developed. •The Q Exactive provides similar sensitivity and enhanced selectivity compared to QqQ. -- Abstract: Sensitive and unequivocal determination of analytes/contaminants in complex matrices is a challenge in the field of food safety control. In this study, various acquisition modes (Full MS/AIF, Full MS + tMS/MS, Full MS/dd MS/MS and tSIM/ddMS/MS) and parameters of a quadrupole–orbitrap hybrid mass spectrometer (Q Exactive) were studied in detail. One of the main conclusions has been that, reducing the scan range for Full MS (using the quadrupole) and targeted modes give higher signal-to-noise (S/N) ratios and thereby better detection limits for analytes in matrix. The use of Q Exactive in a complex case, for the confirmatory analysis of hormones in animal urine is presented. A targeted SIM data dependent MS/MS (tSIM/ddMS/MS) acquisition method for determination of eight synthetic hormones (trenbolone, 17α ethinylestradiol, zeranol, stanozolol, dienestrol, diethylstilbestrol, hexestrol, taleranol) and a naturally occurring hormone (zearalenone) in animal urine were optimized to have sensitive precursors from targeted SIM mode and trigger MS/MS scans over the entire chromatograph peak. The method was validated according to EC/657/2002. CCα (decision limit) for the analytes ranged between 0.11 μg L{sup −1} and 0.69 μg L{sup −1} and CCβ (detection capability) ranged between 0.29 μg L{sup −1} and 0.90 μg L{sup −1}.

  20. Generation of a fluorescent transgenic zebrafish for detection of environmental estrogens

    Chen Hao; Hu Jingying; Yang Jian; Wang Yuexiang; Xu Hui; Jiang Qiu; Gong Yuebo; Gu Yinliang [Department of Molecular Genetics, Shanghai Medical School and Key Laboratory of Molecular Medicine, Ministry of Education, Fudan University, 200032 (China); Song Houyan, E-mail: [Department of Molecular Genetics, Shanghai Medical School and Key Laboratory of Molecular Medicine, Ministry of Education, Fudan University, 200032 (China)


    To establish a novel in vivo test system for rapid detection of environmental estrogens, an ere-zvtg1: gfp transgenic zebrafish line has been generated. In this transgenic line, under control conditions, GFP was exclusively expressed in the liver of mature adult female fish. Male and larval transgenic fish did not express GFP but could be induced to express GFP in the liver after exposure to 17-{alpha}-ethynylestradiol (EE{sub 2}). Concurrent accumulation of zvtg1 and gfp mRNAs in embryos and larvae after EE{sub 2} exposure was observed, which indicated that the expression of gfp transgene was driven by the zvtg1 promoter. Green fluorescence was first observed in the liver at 53, 74, 100 or 131 h post-fertilization (hpf) after exposure to 100, 10, 1 or 0.1 ng/L EE{sub 2} from 1 to 2 cell stage, respectively. As for mature male transgenic zebrafish, green fluorescence was observed after exposure to 100, 10, 1 or 0.1 ng/L EE{sub 2} for 2, 3, 4 or 7 days, respectively; as for mature female, fluorescence was increased after exposure to relatively high concentrations of EE{sub 2} (10 and 100 ng/L). Green fluorescence in the liver was increased with prolonging of exposure time and was repeatedly induced after removal and re-addition of EE{sub 2}. We also demonstrated that GFP expression could be induced by other estrogenic compounds, including {beta}-estradiol (E{sub 2}, 0.1 {mu}g/L), cadmium chloride (CdCl{sub 2}, 10 {mu}g/L), zearalenone (50 {mu}g/L), estriol (E{sub 3}, 1 {mu}g/L), diethylstilbestrol (DES, 50 ng/L) bisphenol A (BPA, 1 mg/L) but not by weakly estrogenic compounds such as nonylphenol (NP, up to 10 mg/L), or non-estrogenic steroid hormones such as progesterone (up to 100 mg/L) and 17-hydroxysteroid (up to 50 mg/L). These data suggest the transgenic zebrafish is sensitive and specific for detection of estrogenic compounds. Because the observed-effect concentrations are as low as those of environment and the observed-effect exposure times are very short

  1. Estrogen modification of human glutamate dehydrogenases is linked to enzyme activation state.

    Borompokas, Nikolas; Papachatzaki, Maria-Martha; Kanavouras, Konstantinos; Mastorodemos, Vasileios; Zaganas, Ioannis; Spanaki, Cleanthe; Plaitakis, Andreas


    Mammalian glutamate dehydrogenase (GDH) is a housekeeping enzyme central to the metabolism of glutamate. Its activity is potently inhibited by GTP (IC(50) = 0.1-0.3 μM) and thought to be controlled by the need of the cell in ATP. Estrogens are also known to inhibit mammalian GDH, but at relatively high concentrations. Because, in addition to this housekeeping human (h) GDH1, humans have acquired via a duplication event an hGDH2 isoform expressed in human cortical astrocytes, we tested here the interaction of estrogens with the two human isoenzymes. The results showed that, under base-line conditions, diethylstilbestrol potently inhibited hGDH2 (IC(50) = 0.08 ± 0.01 μM) and with ∼18-fold lower affinity hGDH1 (IC(50) = 1.67 ± 0.06 μM; p < 0.001). Similarly, 17β-estradiol showed a ∼18-fold higher affinity for hGDH2 (IC(50) = 1.53 ± 0.24 μM) than for hGDH1 (IC(50) = 26.94 ± 1.07 μM; p < 0.001). Also, estriol and progesterone were more potent inhibitors of hGDH2 than hGDH1. Structure/function analyses revealed that the evolutionary R443S substitution, which confers low basal activity, was largely responsible for sensitivity of hGDH2 to estrogens. Inhibition of both human GDHs by estrogens was inversely related to their state of activation induced by ADP, with the slope of this correlation being steeper for hGDH2 than for hGDH1. Also, the study of hGDH1 and hGDH2 mutants displaying different states of activation revealed that the affinity of estrogen for these enzymes correlated inversely (R = 0.99; p = 0.0001) with basal catalytic activity. Because astrocytes are known to synthesize estrogens, these hormones, by interacting potently with hGDH2 in its closed state, may contribute to regulation of glutamate metabolism in brain.

  2. The effects of in utero bisphenol A exposure on reproductive capacity in several generations of mice

    Ziv-Gal, Ayelet, E-mail:; Wang, Wei, E-mail:; Zhou, Changqing, E-mail:; Flaws, Jodi A., E-mail:


    In utero bisphenol A (BPA) exposure affects reproductive function in the first generation (F1) of mice; however, not many studies have examined the reproductive effects of BPA exposure on subsequent generations. In this study, pregnant mice (F0) were orally dosed with vehicle, BPA (0.5, 20, and 50 μg/kg/day) or diethylstilbestrol (DES; 0.05 μg/kg/day) daily from gestation day 11 until birth. F1 females were used to generate the F2 generation, and F2 females were used to generate the F3 generation. Breeding studies at the ages of 3, 6, and 9 months were conducted to evaluate reproductive capacity over time. Further, studies were conducted to evaluate pubertal onset, litter size, and percentage of dead pups; and to calculate pregnancy rate, and mating, fertility, and gestational indices. The results indicate that BPA exposure (0.5 and 50 μg/kg/day) significantly delayed the age at vaginal opening in the F3 generation compared to vehicle control. Both DES (0.05 μg/kg/day) and BPA (50 μg/kg/day) significantly delayed the age at first estrus in the F3 generation compared to vehicle control. BPA exposure reduced gestational index in the F1 and F2 generations compared to control. Further, BPA exposure (0.5 μg/kg/day) compromised the fertility index in the F3 generation compared to control. Finally, in utero BPA exposure reduced the ability of female mice to maintain pregnancies as they aged. Collectively, these data suggest that BPA exposure affects reproductive function in female mice and that some effects may be transgenerational in nature. - Highlights: • In utero BPA delayed vaginal opening in the F3 generation compared to control. • In utero BPA delayed estrus in the F3 generation compared to control. • In utero BPA reduced the ability of F1 and F2 female mice to maintain pregnancies. • In utero BPA compromised the ability of F3 female mice to become pregnant. • Some effects of in utero BPA may be transgenerational in nature.

  3. Mammary tumorigenesis by radiation and its prevention

    Onoda, Makoto; Suzuki, Keiko; Inano, Hiroshi [National Inst. of Radiological Sciences, Chiba (Japan)


    Since the breast cancer in women emerged as an important risk associated with exposure to ionizing radiation, we have investigated to clarify the relationship between the induction of mammary tumors by irradiation and the developmental stage of the mammary glands that regulated by the action of endocrine hormones. Besides the radiation, epidemiological studies showed that the process of biosynthesis/metabolism of steroid hormones and hyperlipidemia may be associated with an increased risk of mammary carcinogenesis. In this context, we have undertaken investigations to evaluate the anti-carcinogenic activities of dehydroepiandrosterone (DHEA), a major secretory steroid of the adrenal glands, bezafibrate (BEZF), an anti-hyperlipidemic drug derived from clofibrate, and simvastatin (SIMV), a prodrug of a specific inhibitor of 3-hydroxy-3-methylglutaryl-CoA reductase, against diethylstilbestrol (DES)-dependent promotion/progression of rat mammary tumors initiated by {gamma}-rays. Pregnant Wistar-MS rats received whole-body irradiation with 2.6 Gy of {gamma}-rays from a {sup 60}Co source at day-20 of pregnancy. The mother rats were fed a diet containing either 0.6% DHEA, 0.15% BEZF or 0.03% SIMV beginning immediately after weaning. They were then implanted subcutaneously with a pellet of DES (3 mg/pellet) in the interscapular area 30 days after termination of nursing and were observed for 1 year for detection of palpable mammary tumors starting from the time of pellet implantation. The administration of dietary DHEA, BEZF or SIMV together with DES implantation in rats irradiated in late pregnancy significantly decreased the total incidence of mammary tumors to 35%, 27% and 36%, respectively, for the 1 year period, while higher tumor incidence (96%, 90% and 88%) was observed in rats fed controldiet. However, neither the number of mammary tumors per tumor-bearing rat nor the latency period in the drug treated groups was different from that observed in the control group

  4. Necessity of re-evaluation of estramustine phosphate sodium (EMP) as a treatment option for first-line monotherapy in advanced prostate cancer.

    Kitamura, T


    Estramustine phosphate sodium (EMP) was first introduced in the early 1970s for the treatment of prostate cancer, when EMP was supposed to have the dual effect of estrogenic activity and cytotoxicity. For the following decades, it was used mainly in hormone-refractory cases, with a conventional dosage of 4-9 capsules/day, which showed a 30-35% objective response rate. However, a very limited number of cases have been reported that used EMP as a first-line monotherapy in the conventional dosage. One study showed a response rate of 82%, which is at least as effective as conventional estrogen (diethylstilbestrol; DES) monotherapy. Nevertheless, EMP was almost abandoned for the treatment of prostate cancer because of severe adverse side-effects, especially in the cardiovascular system and gastrointestinal tract. Recently, two facts have become evident. First, EMP interferes with cellular microtubule dynamics but does not show alkylating effects. Second, EMP is able to produce a complex with calcium when dairy products are taken concomitantly with EMP, resulting in a decrease in the absorption rate of EMP from the gut. Many clinical trials have been undertaken without warning against concomitant dairy product intake since the introduction of EMP. This fact will jeopardize almost all the clinical trials performed before 1990. It is considered that response rates have been underestimated and better results could have been obtained because side-effects decrease dose-dependently. Low-dose EMP monotherapy (2 capsules/day) has been performed infrequently in previously untreated advanced prostate cancer. The only large trial by the European Organization for Research and Treatment of Cancer in 1984 was biased in selecting patients. Nevertheless, the response rate of EMP is comparable to that of DES. In this study, the adverse side-effects of EMP were less than that of DES. Recently, a study was conducted at the University of Tokyo of 11 patients with advanced prostate cancer on

  5. Full automation of solid-phase microextraction/on-fiber derivatization for simultaneous determination of endocrine-disrupting chemicals and steroid hormones by gas chromatography-mass spectrometry.

    Yang, Lihua; Lan, Chongyu; Liu, Hongtao; Dong, Jun; Luan, Tiangang


    A fully automated method using direct immersion solid-phase microextraction (DI-SPME) and headspace on-fiber silylation for simultaneous determinations of exogenous endocrine-disrupting chemicals (EDCs) and endogenous steroid hormones in environmental aqueous and biological samples by gas chromatography-mass spectrometry (GC-MS) was developed and compared to a previously reported manual method. Three EDCs and five endocrine steroid hormones were selected to evaluate this method. The extraction and derivatization time, ion strength, pH, incubation temperature, sample volume, and extraction solvent were optimized. Satisfactory results in pure water were obtained in terms of linearity of calibration curve (R2=0.9932-1.0000), dynamic range (3 orders of magnitude), precision (4-9% RSD), as well as LOD (0.001-0.124 microg L(-1)) and LOQ (0.004-0.413 microg L(-1)), respectively. These results were similar to those obtained using a manual method, and moreover, the precision was improved. This new automated method has been applied to the determinations of target compounds in real samples used in our previous study on a manual SPME method. Exogenous octylphenol (OP), technical grade nonylphenol (t-NP), and diethylstilbestrol (DES) were at 0.13, 5.03, and 0.02 microg L(-1) in river water and 3.76, 13.25, and 0.10 microg L(-1) in fish serum, respectively. Natural steroid hormones estrone (E1), 17beta-estradiol (E2), and testosterone (T) were at 0.19, 0.11, and 6.22 microg L(-1) in river water; and in female fish serum E1, E2, and pregnenolone (PREG) were at 1.37, 1.95, and 6.25 microg L(-1), respectively. These results were confirmed by the manual method. The developed fully automated SPME and on-fiber silylation procedures showed satisfactory applications in environmental analysis and the performances show improved precision and a reduced analysis time compared to the manual method.

  6. Effects of 2,3,7,8-TCDD and benzo[a]pyrene on modulating vitellogenin expression in primary culture of crucian carp (Carassius auratus) hepatocytes

    LIANG Yong; C. K. C. Wong; XU Ying; M. H. Wong


    Vitellogenin (Vtg) is the precursor of yolk protein. Its expression and secretion are estrogen-regulated and are crucial for oocyte maturation. An in vitro xenoestrogen screening model was established by measuring Vtg induction in cultured primary hepatocytes from crucian carp. Vtg production was detected by biotin-avidin sandwich ELISA method while Vtg and cytochrome P4501A1 (CYP1A1) mRNA induction were measured by semi- quantitative PCR-primer dropping technique. Vtg and Vtg mRNA were dose-dependently induced by diethylstilbestrol (DES, 0.2-200 ng/mL) in hepatocytes of crucian carp. Co-treatment of the DES-induced hepatocytes with either 2,3,7,8-TCDD (TCDD, 0.1-4 pg/mL) or benzo[a]pyrene (B[a]P, 5-1000 ng/mL) resulted in a reduction of Vtg production and an increment of CYP1A1 mRNA expression both in a dose dependent manner, indicating the anti-estro-genic effects of the compounds. However, at lower tested concentrations, TCDD (0.1, 0.2 pg/mL), B[a]P (5 ng/mL) seemed to have a potentiating effect on Vtg expression and secretion, although by their own these compounds had no observable estrogenic effect on Vtg induction. Tamoxifen (a selective estrogen receptor modulators, 1 nmol/L-1 μmol/L), and β-naphtho-flavone (β-NF, an aryl hydrocarbon receptor inducing compounds, 2.5-1000 ng/mL) also were employed to study the possible interactions in DES-induced Vtg expression. In co-treatment of the DES-induced hepatocytes with β-NF or tamoxifen, the decrease in Vtg production did parallel induction of CYP1A1 for β-NF, but tamoxifen inhibited Vtg induction did not parallel induced CYP1A1 expression in all test concentrations. On the contrary, it was found that in co-treatment of the TCDD-induced hepatocytes with DES, TCDD induced CYP1A1 mRNA production was inhibited by DES also. These results implicated a possible cross talk between estrogen receptor- and aryl hydrocarbon receptor-mediated pathways in the hepatocytes.

  7. INSL-3 is expressed in human hyperplastic and neoplastic thyrocytes.

    Hombach-Klonisch, Sabine; Hoang-Vu, Cuong; Kehlen, Astrid; Hinze, Raoul; Holzhausen, Hans-Jürgen; Weber, Ekkehard; Fischer, Bernd; Dralle, Henning; Klonisch, Thomas


    The insulin-like hormone INSL-3, also named relaxin-like factor (RLF) or Leydig-derived insulin-like peptide (LEY-IL), is expressed in various reproductive tissues and is regarded a marker of differentiation in human testicular Leydig cells. Recently, we have identified differential expression of human INSL-3 in neoplastic Leydig cells and mammary epithelial cells suggesting an involvement of INSL-3 in tumor biology. Here we have investigated the expression of INSL-3 in human thyroid carcinoma cell lines and in the human thyroid gland which has been shown to express transcripts for the G protein coupled INSL-3 receptor LGR8. When we determined the expression of INSL-3 in eight human thyroid carcinoma cell lines, a novel INSL-3 splice variant containing a 95 bp out-of-frame insertion at the beginning of exon II of the INSL-3 gene was discovered. Treatment of the human anaplastic thyroid carcinoma cell line 8505C with diethylstilbestrol (DES) caused a significant dose-dependent transcriptional down-regulation of INSL-3 and a marked up-regulation of LGR8. Employing in situ hybridization to detect INSL-3 transcripts and specific rabbit antisera against the INSL-3 proteins, both INSL-3 isoforms were detected in patients with Graves' disease (n=10), follicular carcinomas (FTC; n=12), papillary carcinomas (PTC; n=9) and undifferentiated anaplastic carcinomas (UTC; n=15). By contrast, thyrocytes of all 15 benign goiter tissues studied were devoid of both INSL-3 isoforms, mRNA and protein. Our data indicate that INSL-3 hormone is up-regulated in hyperplastic and neoplastic human thyrocytes suggesting that the INSL-3 isoforms may serve as additional markers for hyperplastic and neoplastic human thyrocytes. In the anaplastic thyroid carcinoma cell line 8505C, the regulation of both INSL-3 and LGR8 by estrogen may be the first indication of a novel hormonally responsive, auto-/paracrine INSL-3 LGR8 ligand receptor system active in human thyroid carcinoma cells.

  8. The pancreas is altered by in utero androgen exposure: implications for clinical conditions such as polycystic ovary syndrome (PCOS.

    Mick Rae

    Full Text Available Using an ovine model of polycystic ovary syndrome (PCOS, (pregnant ewes injected with testosterone propionate (TP (100 mg twice weekly from day (d62 to d102 of d147 gestation (maternal injection - MI-TP, we previously reported female offspring with normal glucose tolerance but hyperinsulinemia. We therefore examined insulin signalling and pancreatic morphology in these offspring using quantitative (Q RT-PCR and western blotting. In addition the fetal pancreatic responses to MI-TP, and androgenic and estrogenic contributions to such responses (direct fetal injection (FI of TP (20 mg or diethylstilbestrol (DES (20 mg at d62 and d82 gestation were assessed at d90 gestation. Fetal plasma was assayed for insulin, testosterone and estradiol, pancreatic tissue was cultured, and expression of key β-cell developmental genes was assessed by QRT-PCR. In female d62MI-TP offspring insulin signalling was unaltered but there was a pancreatic phenotype with increased numbers of β-cells (P<0.05. The fetal pancreas expressed androgen receptors in islets and genes involved in β-cell development and function (PDX1, IGF1R, INSR and INS were up-regulated in female fetuses after d62MI-TP treatment (P<0.05-0.01. In addition the d62MI-TP pancreas showed increased insulin secretion under euglycaemic conditions (P<0.05 in vitro. The same effects were not seen in the male fetal pancreas or when MI-TP was started at d30, before the male programming window. As d62MI-TP increased both fetal plasma testosterone (P<0.05 and estradiol concentrations (P<0.05 we assessed the relative contribution of androgens and estrogens. FI-TP (commencing d62 (not FI-DES treatment caused elevated basal insulin secretion in vitro and the genes altered by d62MI-TP treatment were similarly altered by FI-TP but not FI-DES. In conclusion, androgen over-exposure alters fetal pancreatic development and β-cell numbers in offspring. These data suggest that that there may be a primary pancreatic

  9. Resveratrol promotes expression of SIRT1 and StAR in rat ovarian granulosa cells: an implicative role of SIRT1 in the ovary

    Morita Yoshihiro


    Full Text Available Abstract Background Resveratrol is a natural polyphenolic compound known for its beneficial effects on energy homeostasis, and it also has multiple properties, including anti-oxidant, anti-inflammatory, and anti-tumor activities. Recently, silent information regulator genes (Sirtuins have been identified as targets of resveratrol. Sirtuin 1 (SIRT1, originally found as an NAD+-dependent histone deacetylase, is a principal modulator of pathways downstream of calorie restriction, and the activation of SIRT1 ameliorates glucose homeostasis and insulin sensitivity. To date, the presence and physiological role of SIRT1 in the ovary are not known. Here we found that SIRT1 was localized in granulosa cells of the human ovary. Methods The physiological roles of resveratrol and SIRT1 in the ovary were analyzed. Immunohistochemistry was performed to localize the SIRT1 expression. SIRT1 protein expression of cultured cells and luteinized human granulosa cells was investigated by Western blot. Rat granulosa cells were obtained from diethylstilbestrol treated rats. The cells were treated with increasing doses of resveratrol, and subsequently harvested to determine mRNA levels and protein levels. Cell viability was tested by MTS assay. Cellular apoptosis was analyzed by caspase 3/7 activity test and Hoechst 33342 staining. Results SIRT1 protein was expressed in the human ovarian tissues and human luteinized granulosa cells. We demonstrated that resveratrol exhibited a potent concentration-dependent inhibition of rat granulosa cells viability. However, resveratrol-induced inhibition of rat granulosa cells viability is independent of apoptosis signal. Resveratrol increased mRNA levels of SIRT1, LH receptor, StAR, and P450 aromatase, while mRNA levels of FSH receptor remained unchanged. Western blot analysis was consistent with the results of quantitative real-time RT-PCR assay. In addition, progesterone secretion was induced by the treatment of resveratrol

  10. Efficacy and tolerability of high dose "ethinylestradiol" in post-menopausal advanced breast cancer patients heavily pre-treated with endocrine agents

    Robertson John FR


    HDEs (such as diethylstilbestrol or chemotherapy.

  11. Evaluation of mechanisms inducing thyroid toxicity and the ability of the enhanced OECD Test Guideline 407 to detect these changes.

    Cunha, G Coelho-Palermo; van Ravenzwaay, B


    The OECD has developed an "enhanced Test Guideline 407" (TG 407) protocol for detecting endocrine effects during the course of a 28-day testing scheme. This protocol has gone through a validation process with (anti)estrogenic and (anti)androgenic compounds and substances that affect the thyroid (thyroxine and propylthiouracil). This review investigates whether a 28-day testing scheme would show up alterations in the thyroid-related parameters of the "enhanced TG 407" (T3, T4, TSH, thyroid weight and histopathology), irrespective of the mode of action. For each mode of action, a generally accepted reference chemical was selected and an in-depth literature survey was carried out, and the chemical was evaluated for treatment-related changes of thyroid-dependent parameters. The following model chemicals were selected: ion perchlorate, blockage of iodine uptake; propylthiouracil, inhibition of thyroid hormone synthesis; excess of iodine, blockage of thyroid hormone release; pyrazole, thyroid cytotoxicity; minocycline, thyroid pigmentation; amiodarone, inhibition of TSH synthesis; diethylstilbestrol, competition for thyroid hormone binding globulin; selenium-deficient diet, inhibition of thyroxine deiodination; FD&C Red No. 3, inhibition of peripheral 5'-deiodinase; cadmium, lipid peroxidation; phenobarbital, increase in thyroxine conjugation and biliary excretion; temelastine, thyroxine accumulation. Test data for treatments lasting approximately one month were available for most of these model chemicals, and these demonstrated the expected thyroid-related changes. Thus, it can be concluded that a 28-day testing scheme allows for the detection of thyroid-disrupting chemicals. The literature data also were evaluated according to whether preference can be given to any of the thyroid-related parameters (thyroid/pituitary hormones, thyroid weight and histopathology) with regard to dose-related sensitivities. Due to different study designs (such as treatment duration

  12. Analysis of estrogens residues in milk by gas chromatographic-tandem mass spectrometry%GC-MS法测定生鲜乳中雌激素类药物残留量

    王丽娜; 田晓玲; 陈玉艳


    本研究建立了测定生鲜乳中己烯雌酚(diethylstilbestrol)、雌二醇(estradiol)、炔雌醇(ethi-nylestradiol)、戊酸雌二醇(estradiol valerate)残留量的GC- MS法,为控制食源性动物食品的质量安全提供依据。本试验的样品经乙腈和乙酸乙酯提取,HLB固相萃取小柱净化,70℃衍生后在EI源SIM模式下进样分析。试验结果显示,己烯雌酚、雌二醇、炔雌醇含量在2.0~100.0μg/kg内、戊酸雌二醇含量在4.0~200.0μg/kg内线性关系良好,己烯雌酚、雌二醇、炔雌醇、戊酸雌二醇相关系数分别为0.9991、0.9987、0.9978、0.9983,平均回收率(n=6)为74.2%~85.8%(RSD为1.9%~9.2%)。该方法能够为测定生鲜乳中这4种雌激素类药物残留量提供可靠依据。%This research to develop and validate a rapid sensitive and specific meth-od for quantitative analytes of estrogens residues(DES, Diethylstilbestrol: E2, Es-tradiol: EE2, Ethynylestradiol: EV, Estradiol Valerate)in milk by gas chromatograph-ic-tandem mass spectrometry and be applied to estrogens residues in milk studies. The 4 kinds of components were extracted from homogenized samples with ethyl acetate fol-lowed by solid-phase extraction with HLB column and trimethylsilylation at 70 ℃ for 40 minutes, then the GC-MS method in selected ion monitoring mode(SIM)was used to an-alyze them. A good linearity was achieved with the coefficient r2 > 0.997 8(n =2). The average recovery of estrogens was more than 74.2% with RSD lower than 9.2%(n = 9). The method is sensitive, accurate, rapid, and suitable for the analysis estrogens resi-dues in milk.

  13. Environmental signaling: from environmental estrogens to endocrine-disrupting chemicals and beyond.

    McLachlan, J A


    The landmark report (Herbst et al. 1971) linking prenatal treatment with a synthetic estrogen, diethylstilbestrol (DES), to cancer at puberty in women whose mothers took the drug while pregnant ushered in an era of research on delayed effects of such exposures on functional outcomes in offspring. An animal model developed in our laboratory at the National Institute of Environmental Health Sciences confirmed that DES was the carcinogen and exposure to DES caused, as well, functional alterations in the reproductive, endocrine, and immune systems of male and female mice treated in utero. DES was also being used in agriculture and we discovered, at the first meeting on Estrogens in the Environment in 1979 (Estrogens in the Environment, 1980), that many environmental contaminants were also estrogenic. Many laboratories sought to discern the basis for estrogenicity in environmental chemicals and to discover other hormonally active xenobiotics. Our laboratory elucidated how DES and other estrogenic compounds worked by altering differentiation through epigenetic gene imprinting, helping explain the transgenerational effects found in mice and humans. At the Wingspread Conference on the Human-Wildlife Connection in 1991 (Advances in Modern Environmental Toxicology, 1992), we learned that environmental disruption of the endocrine system occurred in many species and phyla, and the term endocrine disruption was introduced. Further findings of transgenerational effects of environmental agents that mimicked or blocked various reproductive hormones and the ubiquity of environmental signals, such as bisphenol A increased concern for human and ecological health. Scientists began to look at other endocrine system aspects, such as cardiovascular and immune function, and other nuclear receptors, with important observations regarding obesity and metabolism. Laboratories, such as ours, are now using stem cells to try to understand the mechanisms by which various environmental signals

  14. The relevance of chemical interactions with CYP17 enzyme activity: Assessment using a novel in vitro assay

    Roelofs, Maarke J.E., E-mail: [Endocrine Toxicology Research Group, Institute for Risk Assessment Sciences (IRAS), Utrecht University, P.O. Box 80.177, NL-3508 TD Utrecht (Netherlands); Center for Health Protection, National Institute for Public Health and the Environment (RIVM), P.O. Box 1, 3720 BA Bilthoven (Netherlands); Piersma, Aldert H. [Endocrine Toxicology Research Group, Institute for Risk Assessment Sciences (IRAS), Utrecht University, P.O. Box 80.177, NL-3508 TD Utrecht (Netherlands); Center for Health Protection, National Institute for Public Health and the Environment (RIVM), P.O. Box 1, 3720 BA Bilthoven (Netherlands); Berg, Martin van den; Duursen, Majorie B.M. van [Endocrine Toxicology Research Group, Institute for Risk Assessment Sciences (IRAS), Utrecht University, P.O. Box 80.177, NL-3508 TD Utrecht (Netherlands)


    The steroidogenic cytochrome P450 17 (CYP17) enzyme produces dehydroepiandrosterone (DHEA), which is the most abundant circulating endogenous sex steroid precursor. DHEA plays a key role in e.g. sexual functioning and development. To date, no rapid screening assay for effects on CYP17 is available. In this study, a novel assay using porcine adrenal cortex microsomes (PACMs) was described. Effects of twenty-eight suggested endocrine disrupting compounds (EDCs) on CYP17 activity were compared with effects in the US EPA validated H295R (human adrenocorticocarcinoma cell line) steroidogenesis assay. In the PACM assay DHEA production was higher compared with the H295R assay (4.4 versus 2.2 nmol/h/mg protein). To determine the additional value of a CYP17 assay, all compounds were also tested for interaction with CYP19 (aromatase) using human placental microsomes (HPMs) and H295R cells. 62.5% of the compounds showed enzyme inhibition in at least one of the microsomal assays. Only the cAMP inducer forskolin induced CYP17 activity, while CYP19 was induced by four test compounds in the H295R assay. These effects remained unnoticed in the PACM and HPM assays. Diethylstilbestrol and tetrabromobisphenol A inhibited CYP17 but not CYP19 activity, indicating different mechanisms for the inhibition of these enzymes. From our results it becomes apparent that CYP17 can be a target for EDCs and that this interaction differs from interactions with CYP19. Our data strongly suggest that research attention should focus on validating a specific assay for CYP17 activity, such as the PACM assay, that can be included in the EDC screening battery. - Highlights: ► DHEA, produced by CYP17, plays a key role in sexual functioning and development. ► No rapid screening assay for effects on CYP17 is available yet. ► A novel assay using porcine adrenal cortex microsomes (PACMs) was described. ► Endocrine disrupting compounds (EDCs) targeting CYP17 interact differently with CYP19. ► A

  15. Regulation of progesterone receptor messenger ribonucleic acid in the rat medial preoptic nucleus by estrogenic and antiestrogenic compounds: an in situ hybridization study.

    Shughrue, P J; Lane, M V; Merchenthaler, I


    Progesterone receptor (PR) messenger RNA (mRNA) is concentrated in neurons of the preoptic area and other regions of the rat hypothalamus where it is colocalized with the estrogen receptor and regulated by changes in the steroid hormonal milieu. To date, little is known about the regulation of PR mRNA by estrogens and whether antiestrogenic compounds are capable of modulating its expression. The present studies used in situ hybridization to ascertain the time course of PR mRNA regulation in the medial preoptic nucleus by 17beta-estradiol, determine the effective dose required to elicit a response, and compare the efficacy of 17beta-estradiol with a variety of estrogenic or antiestrogenic compounds. The first series of studies revealed that the treatment of ovariectomized rats with 17beta-estradiol resulted in an increase in PR expression within 2 h, after which it remained elevated until 10 h postinjection and then returned to baseline levels. When ovariectomized rats were injected with 25-1000 ng/kg of 17beta-estradiol and euthanized 6 h later, a dose-dependent increase in the level of PR mRNA was observed, with a maximal response at 1000 ng/kg and an EC50 of 93.5 ng/kg. Subsequent studies evaluated the efficacy of a variety of estrogenic and antiestrogenic compounds in the rat preoptic nucleus. 17Beta-estradiol, diethylstilbestrol, and 17alpha-estradiol all significantly increased the level of PR mRNA, although the degree of induction varied with each compound. The injection of tamoxifen, raloxifene, toremifene, droloxifene, clomiphene, GW 5638, or ICI 182,780 had no significant estrogenic effect on PR gene expression at the dose evaluated. In contrast, when tamoxifen or raloxifene, but not ICI 182,780, was administered in the antagonist mode, a significant dose-related decrease in the estradiol-induced level of PR mRNA was seen in the preoptic area. The results of these studies clearly demonstrate that PR mRNA expression in the rat preoptic area is rapidly

  16. Growth hormone secretion and clearance rates in growing beef steers implanted with estrogenic anabolic compounds.

    Gopinath, R; Kitts, W D


    The effect of estrogenic anabolic compounds on the kinetic parameters of metabolism of growth hormone (GH) was studied in growing beef steers. Twenty-four beef steers were randomly placed into four groups and assigned to one of the following four treatment groups: zeranol, diethylstilbestrol (DES), Synovex-S and an unimplanted control. GH metabolism was studied from eight steers on day 20 following the implantation of anabolic compounds. The animals were rapidly injected with a solution of bGH (NIH-GH-B18) and the disappearance of injected GH from the plasma was monitored up to 120 min following the injection. The plasma GH disappearance curve displayed an initial rapid phase lasting 5 min and a slow disappearance phase lasting 42 min; the fractional turnover rate from the two compartments were 0.167 and 0.017 min-1, respectively. The average volume of distribution of GH in steers was 6% of the body weight. Mean values of metabolic clearance and secretion rates of GH in steers were 21 liters/h and 252 micrograms/h or 74.5 ml/kg/h and 0.91 microgram/kg/h, respectively. Steers implanted with anabolic compounds gained more rapidly (P less than 0.05) than the controls. Plasma basal GH concentration appeared to be higher in all the implanted than in the control steers. The secretion rate of GH was increased (P less than 0.05) in steers implanted with anabolic compounds when compared to control steers. The secretion rate (microgram/kg/h) was about 96% (P less than 0.05), 107% (P less than 0.05) and 81% (P less than 0.05) higher in steers implanted with DES, zeranol and Synovex-S, respectively, than in the control steers. All the compounds studied were equally effective in increasing the secretion of GH on day 20 following their implantation. Metabolic clearance rate of GH was not affected by anabolic compound implantation in steers. There was, however, a slight reduction in metabolic clearance rate due to DES and a slight elevation due to zeranol and Synovex-S when

  17. 同位素稀释-超高效液相色谱-串联质谱法同时测定精油中的7种雌性激素%Simultaneous determination of 7 female sex hormones in essential oil by high performance liquid chromatography- tandem mass spectrometry with isotope dilution

    黄百芬; 韩铮; 徐小民; 蔡增轩; 姜维; 任一平


    建立了采用同位素稀释-超高效液相色谱-串联质谱同时快速测定精油中7种雌性激素(雌三醇、雌二醇、雌酮、炔雌醇、己二烯雌酚、己烷雌酚、己烯雌酚)的方法.样品中雌性激素用乙酸乙酯-正己烷(2∶98, v/v)溶液提取后,经硅胶固相萃取小柱净化,通过ACQUITY UPLCTM BEH SHELD RP18色谱柱(100 mm×2.1 mm, 1.7 μm)、以水-乙腈作流动相梯度洗脱对7种雌性激素进行分离,采用串联质谱在负离子扫描方式下通过多反应监测(MRM)模式进行定性定量分析.以雌三醇-D3、雌二醇-D3、己烯雌酚-D6为内标,有效减少了样品基质的影响.该方法对精油中7种雌激素的检出限(LOD)为0.3~7 μg/kg,定量限(LOQ)为1~20 μg/kg.待测物与内标物定量离子的峰面积比值与待测物的质量浓度在20~500 μg/L范围内呈良好的线性关系,相关系数(r2)均大于0.997;在20~500 μg/kg 范围内3个水平的加标平均回收率为88.5% ~114.8% ,日内精密度(以相对标准偏差计)(n=6)为4.8% ~18.9% .应用该方法对浙江杭州地区不同超市或美容院随机采集的12份精油样品进行测定的结果显示,有1份样品含有雌二醇和雌酮,其余11份样品均未检出雌性激素.%A reliable ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous determination of 7 female sex hormones ( estriol,estradiol, estrone, ethinyloestradiol, dienestrol, hexestrol, diethylstilbestrol) in essential oil was developed. The sample was extracted by ethylacetate-normal hexane solution (2: 98, v/v)and the extract was purified by a silica solid phase extraction-based clean-up column. Then,the analytes were separated on an ACQUITY UPLC BEH SHELD RP18 column ( 100 mm × 2.1 mm, 1.7 μm) in gradient elution with the mobile phases of water and acetonitrile. The separated compounds were detected with a Waters Xevo TQ MS tandem quadrupole mass spectrometer operated in

  18. Effect of Sierqiwu Particles on the Premature Ovarian of the Rats Serum AMH%四二杞乌颗粒对卵巢早衰模型大鼠血清抗苗勒管激素的影响

    孙夕平; 刘冬岩


    Objective To study the changes of serum the anti-Müllerian hormone (AMH) and sex hormones of Sierqiwu particles on premature ovarian failure (POF) rats induced by Tripterygium wilfordii polyglycoside (TWP), and explore the value of the AMH in the early prediction and diagnosis of premature ovarian failure.Methods 40 12-week-old female Wistar rats were randomly divided into model group (n=30) and blank control group (n=10), the first three groups to build modules.After establishing successfully rat model of premature ovarian failure with TWP in model group,then they were randomly divided into Sierqiwu particles experimental group, diethylstilbestrol control group, model control group ,and then they were respectively given Sierqiwu particle suspension, diethylstilbestrol suspension, same dose saline (control group and blank model control groups) for 2 weeks. Serum anti-Müllerian hormone(AMH), follicle-stimulating hormone(FSH), luteinizing hormone(LH), estradiol (E2) rat ovarian tissue morphology and estrous cyclewere obtained. Result The level of serum AMH ,E2 were significantly lower,and FSH, LH significantly increased in premature ovarian failure compared with bank group control; but with the treatment of Sierqiwu particles, serum AMH, E2, FSH, LH levels closed to bank group control (normal levels), and meanwhile improved rat ovarian tissue morphology and estrous cycle.Conclusion Effect of Sierqiwu Particles on the Premature Ovarian in the Rats is actively and AMH can be used as one of the predictors of premature ovarian failure.%目的:研究四二杞乌颗粒对雷公藤多甙片(TWP)所致的卵巢早衰(POF)模型大鼠血清抗苗勒管激素(AMH)及性激素的水平变化,探讨抗苗勒管激素(AMH)在早期预测和诊断 POF 中的价值。方法40只12周龄健康雌性 Wistar 大鼠随机分为造模组(30只)空白对照组(10只)。造模组成功建立 TWP 所致的大鼠 POF 模型后,将模型组随机分为四二

  19. Relationship between environmental estrogens and hypospadias: a meta-analysis%外源性雌激素与尿道下裂关系的meta分析

    云妙英; 阿德娜依·阿力肯; 秦颖; 赵智慧; 刘琪; 徐刚


    Objective To evaluate the correlation between environmental estrogen and incidence of hypospadias.Methods Medline,Web of Science (ISI),EMBASE,the Cochrane Library,EBSCO Host,Elsevier Science Direct,Chinese Biomedicine (CBMdisc),and China National Knowledge Infrastructure(CNKI) databases,had been searched and secondary manual search for library was also conducted.Case-control and cohort studies,reporting odds ratio (OR) or relative risk (RR) of environmental estrogen exposure before or during pregnancy and their 95% confidence interval (CI) for the occurrence of hypospadias and published from January 1990 to July 2011,were included.Review Manager 5.2 software were used for data analysis.Results Altogether,18 studies were finally recruited including two cohort studies (only diethylstilbestrol was involved) and 16 case-control studies.While the 16 case-control studies showed significant heterogeneity (P=0.002),and the pooled OR (95%CI) calculated by random effect model was 1.35(1.08-1.70).For the three case-control studies on diethylstilbestrol exposured during pregnancy,no heterogeneity was found (P=0.250) and the pooled OR (95%CI) with fixed effect model was 2.12(1.18-3.80).For the three case-control studies on oral contraceptives and the other three on exposure to dichlorodiphenyltrichloroethane (DDT) or DDT-derived materials,the pooled OR (95%CI) were 1.01 (0.81-1.27) and 1.04 (0.61-1.74) respectively with fixed effect model as no heterogeneity was found (P=0.450 and 0.980).There were two case-control studies on phytoestrogens and two on phthalates,and both showed significant heterogeneity (both P=0.020).The pooled OR (95%CI) with random effect model were 1.88 (0.52-6.79) and 1.68 (0.47-5.94),respectively.The pooled OR (95%CI) of case-control studies in Europe (11 studies) and North America(five studies) were 1.51 (1.13-2.03) and 1.13(0.86 1.53) with the random and fixed effect model,separately (heterogeneity test P values were 0.002 and 0

  20. Uterotrophic assay of two concentrations of migrates from each of 23 polystyrenes administered orally (by gavage) to immature female Wistar rats.

    Bachmann, S; Hellwig, J; Jäckh, R; Christian, M S


    The Styrene Steering Committee (SSC) of the European Chemical Industry Council (CEFIC) sponsored this work to address any concern that styrene dimers and trimers that might migrate from polystyrene containers into food could possess some estrogenic activity and thus possibly affect human health. All phases of the study were conducted in conformance with GLP regulations and without knowledge of the oligomer migrates tested. All activities were managed and audited under a third-party contract between the SSC and Argus International. Low and high doses of the styrene oligomer migrates of 23 polystyrene samples [i.e. 9 general purpose polystyrenes (GPPS), 8 high impact polystyrenes (HIPS) and 6 expandable polystyrenes (EPS)] were tested for estrogenicity in an in vivo uterotrophic assay (immature female rat model). This model is considered to be the "gold standard" for use in screening for estrogenic effects because it evaluates both direct and indirect potential effects. The two concentrations of migrates of each of the 23 polystyrenes tested were selected to simulate daily human consumption of a low and high amount of food. Representative dimer and trimer concentrations were obtained in conformance with EEC Council Directives and calculated to be at levels simulating human consumption of 0.5 or 5 kg of food for the GPPS and the HIPS samples and of 0.5 or 3.15 kg of food for the EPS samples, respectively. The study was conducted in a series of three blocks. Each block included concurrent untreated control (negative control), vehicle control (25% ethanol, 20 ml/kg/day) and positive control (diethylstilbestrol-dipropionate, DES-DP, 5 micrograms/kg/day) groups, and low and high doses of each of 7 (1 block) or 8 (2 blocks) polystyrene oligomer migrates. Each group in each block consisted of 10 immature Wistar (Chbb: THOM-SPF) female rats. Beginning when the rats were 22 +/- 1 days of age, each rat was appropriately handled (untreated control group) or administered twice

  1. 17 beta-estradiol-BSA conjugates and 17 beta-estradiol regulate growth plate chondrocytes by common membrane associated mechanisms involving PKC dependent and independent signal transduction.

    Sylvia, V L; Walton, J; Lopez, D; Dean, D D; Boyan, B D; Schwartz, Z


    ]-thymidine incorporation, and the effect was dose-dependent. E(2)-BSA caused time-dependent increases in PKC in RC and GC cells; effects were observed within three minutes in RC cells and within one minute in GC cells. Response to E(2) was more robust in RC cells, whereas in GC cells, E(2) and E(2)-BSA caused a comparable increase in PKC. GDP beta S inhibited the activation of PKC in E(2)-BSA-stimulated RC and GC cells. GTP gamma S increased PKC in E(2)-BSA-stimulated GC cells, but had no effect in E(2)-BSA-stimulated RC cells. The phosphatidylinositol-specific PLC inhibitor U73122 blocked E(2)-BSA-stimulated PKC activity in both RC and GC cells, whereas the phosphatidylcholine-specific PLC inhibitor D609 had no effect. Neither the PLD inhibitor wortmannin nor the phosphatidylinositol 3-kinase inhibitor LY294022 had any effect on E(2)-BSA-stimulated PKC activity in either RC or GC cells. The classical estrogen receptor antagonist ICI 182780 was unable to block the stimulatory effect of E(2)-BSA on PKC. Moreover, the classical receptor agonist diethylstilbestrol (DES) had no effect on PKC, nor did it alter the stimulatory effect of E(2)-BSA. The specificity of the membrane response to E(2) was also demonstrated by showing that the membrane receptor for 1 alpha,25-(OH)(2)D(3) was not involved. These data indicate that the rapid nongenomic effect of E(2)-BSA on PKC activity in RC and GC cells is dependent on G-protein-coupled PLC and support the hypothesis that many of the effects of E(2) involve membrane-associated mechanisms independent of classical estrogen receptors. (c) 2001 Wiley-Liss, Inc.

  2. 析因设计DLLME-SFO同时萃取水中EDCs的应用%Application of Factorial Design to Simultaneous Extraction of EDCs from Water Using DLLME-SFO

    王夏娇; 张琛; 刘建林; 李鱼


    This paper refers to simultaneous extraction of endocrine disrupting chemicals including (EDCs), estriol (E3), bisphenol A (BPA), diethylstilbestrol (DES), estrone (El) and nonylphnol (NP) from water sample using dispersive liquid-liquid micro-extraction based on solidification of floating organic drop (abbr. DLLME-SFO) and the factorial design which was used to optimize extraction process. Regression equations for predicting extraction recovery were established, the effects of factors on DLLME-SFO analyzed and then the optimum conditions obtained. Finally, DLLME-SFO was used to analyze EDCs in the tap water and it was concluded that the regression equations could predict the recovery of extraction of EDCs with a relative deviation range of 0.87%-8.28%.%采用全析因设计对分散液液微萃取-上浮溶剂固化(DLLME-SFO)同时萃取水中五种环境内分泌干扰物包括雌酮(E1)、雌三醇(E3)、双酚A(BPA)、己烯雌酚(DES)和壬基酚(NP)的条件进行优化,建立了预测回归方程,并分析了因素对DLLME-SFO萃取回收率的影响.利用SAS软件分析得到的最优条件为盐浓度(m/v)15%、水样体积3mL、分散剂(甲醇)体积0.5 mL以及萃取剂(十二醇)体积120 μL.在优化萃取条件下,E1、E3、BPA、DES和NP萃取回收率的预测值分别为46.17%、74.38%、79.20%、69.62%和73.05%,实验验证值分别为44.15%、87.76%、77.83%、77.68%和70.01%,实验值与预测值的相对偏差小于10%.将建立的DLLME-SFO方法用于测定实际水样中的目标化合物,结果显示实际水样中E3、BPA、DES、E1和NP同时萃取回收率分别为64.81%~76.18%,88.09%~8.3.15%,68.38%~70.08%,66.37%~68.72%,68.35%~72.80%,相对标准偏差(n=3)在1.47%~8.09%之间.

  3. [Specific toxicological considerations on anabolic agents; transmitted toxicity].

    Ferrando, R; Truhaut, R


    The growth of populations and the spread of urbanization, resulting in new agricultural structures, have entailed a concentration of livestock production and recourse to new techniques. Of some importance among these techniques is the enteric or parenteral administration of substances in very low doses. These substances include anabolic agents, some of which, like many natural feeds, exhibit hormonal activity. They may be divided into two classes: --those of the DES type, synthetic compounds non-existent in the natural state, --natural agents, which are normally distributed throughout the animal and human organism, and hence in food of animal origin---milk, meat, eggs. The compounds belonging to the second class may also be synthesized and the main toxicological consideration is that they then have to meet clear-cut standards of identity and purity. A compound belonging to the first class, diethylstilbestrol (DES), administered to rats in doses as small as 60 mug/kg of feed or even smaller, causes in general lower growth rates as well as alterations in the genital system and reproductive functions. In long-term experiments (12 months) using rats and mice and applying so-called toxicity "de relais" tests, developed and described by the authors, it also appeared that meat from calves in which DES pellets were implanted under normal rearing conditions, inhibits growth and reproduction in mice and rats fed a diet containing 20% of this meat. Studies in which the livers from treated calves constituted 6% of the diet of these two rodent species also led to the conclusion that fertility was impaired in the second reproduction test. The authors also recall cases of vaginal cancer observed in young girls whose mothers had been treated with DES during pregnancy. Compounds belonging to the second class (estradiol-progesterone and estradiol-testosterone) gave no evidence of harmful effects upon rats when mixed with their rations during short and medium-term trials. Similar


    周彩虹; 郭纯; 刘建青; 洪小迪; 柳明; 宁保安; 高志贤


    目的 制备聚甲基丙烯酸(PMAA)反蛋白石光子晶体,观察其在不同环境因素刺激下的反应特性,并初步尝试将己烯雌酚(DES)印迹的PMAA光子晶体应用于检测小分子DES,为此类传感器检测小分子提供新的技术.方法 制备DES印迹的PMAA反蛋白石光子晶体.首先采用垂直沉积法制备二氧化硅蛋白石光子晶体模板,通过向模板中灌入预聚合液聚合形成薄膜,再除去二氧化硅模板和目标分子,得到印迹的反蛋白石光子晶体.在光纤光谱仪下观察pH等环境因素对光子晶体反射峰信号的影响.在体积比1∶1的甲醇和水中检测DES.结果 环境因素溶剂乙醇、离子强度、温度对光子晶体的反射峰信号影响较大,pH对其基本没有影响;印迹光子晶体对DES有明显的吸附反应,检测范围为1~500 ng/ml.结论 制备的DES分子印迹反蛋白石光子晶体对环境因素刺激反应灵敏,并且能够检测微量的DES.%Objective To prepare polymerized methyl acrylic acid ( PMAA) photonic crystal, and to observe its characteristic response to different environmental factors, as well as to try to apply it to detecting diethyl-stilbestrol (DES) ,so as to provide a new method for this kind of sensor to detect small molecules. Methods A colloidal crystal template was prepared from monodisperse SiO2 nanospheres by vertical deposition method first. Then the precursors were infused into the void spaces of the SiO2 templates and aggregated at 60 ℃ for 4 h. The template and the imprinted DES were removed,and then molecularly imprinted photonic polymers were prepared. The effects of pH, concentration of ethanol, temperature, and ionic strength on this photonic crystal property were observed using a fiber-optic spectrometer. Finally, DES in the aqueous solution of ethanol ( 1 : 1) was detected. Results The environmental factors such as ethanol solvent,ionic strength, and temperature markedly affected the reflection peak signal

  5. 胚胎发育与环境激素的影响%Effects of environmental hormone on embryonic development

    胡梦桑; 陈毅斐; 徐营


    背景:自然环境中胚胎发育异常,性行为异常,生物两性化以及种族繁殖数量减少等现象的发生概率升高与环境激素有关.随着环境污染的日趋加重,环境激素对胚胎发育的影响已日益受到重视.目的:分析环境激素对早期胚胎着床、着床前期早期胚胎发育及其对胚胎期胚胎的生殖系统发育的影响及其机制.方法:使用环境激素及己烯雌酚、米非司酮、甲氧滴滴涕、胚胎发育、胚胎早期发育等关键词,检索中国知识资源总库与PubMed数据库中的相关研究论文,分析环境激素对不同时期胚胎发育的影响.结果与结论:纳入45篇符合标准的文献.环境激素广泛存在于日常生活中,种类繁多,作用机制复杂多样,且不同环境激素之间可能还存在协同作用,对分子水平的机制探索还处于初级阶段.控制环境激素生成源,尽量减少人为排放是降低环境激素对人类健康影响的重要措施.%BACKGROUND: Many studies have shown that the increasing occurrence of reproductive abnormalities in wild life may be associated with exposu re to environmental hormone. With the pollution of the environment, effects of environmental hormone on embryonic development have been paid more and more attention.OBJECTIVE: To review the effects and possible mechanism of various environmental hormone on the process of embryo implantation, embryonic development during preimplantation and the development of embryonic reproductive system.METHODS: CNKI database and Pubmed database were searched using key words of "environmental hormone and diethylstilbestrol, mifepristone, methoxychlor, bisphenol A, tetrachlorodibenzo-p-dioxin, embryonic development, early embryonic development", for papers add ressing the effects of environmental hormone on embryonic development.RESULTS AND CONCLUSION: Totally 45 articles were adapted. Common environmental hormones widely spread in daily life in a great variety. Different

  6. Protective effect of Huahong capsule on ovariectomy-induced bone loss in rats%化红胶囊对去势大鼠骨质疏松的影响

    周继刚; 薛冰洁; 曹丹; 周创; 陈茂华; 汪鋆植


    目的:探讨化红胶囊对去势大鼠骨质疏松的影响及作用机制.方法:取3月龄SD雌性大鼠经去势手术或者是假阳性手术后,喂以去大豆特殊饲料7周以诱导骨丢失.经去势手术大鼠进一步分为化红胶囊组(1.0,0.5,0.25 g·kg-1)、模型组、己烯雌酚组,连续给药12周.检测大鼠血清的骨钙素、雌二醇、尿中脱氧吡啶啉含量,以及椎骨末端骨小梁的变化情况.结果:化红胶囊能降低大鼠血清中骨钙素以及尿液中脱氧吡啶啉含量,提高血清中雌二醇含量,与模型组相比,1.0 g·kg-1,0.5 g、kg-1剂量组差异有显著性(P<0、05或P<0.01).骨小梁的形态学观察及数据分析显示,与模型组相比,化红胶囊可以提高骨小梁面积百分比,与模型组相比,1.0,0.5g·kg -1剂量组差异有显著性(P<0.05或p<0.01).结论:化红胶囊可以降低去势大鼠骨转换率和骨吸收程度,提高骨小梁平均面积百分比,是一种潜在的治疗绝经后骨质疏松药物,其作用机制之一可能与化红胶囊雌激素样作用有关.%OBJECTIVE To study the protective effect of Huahong capsule on ovariectomy-induced bone loss in rats and its mechanisms of actions. METHODS Three-month-old female Sprague-Dawley rats were ovariectomized(OVX) and fed on special diet without soybeen for 7 weeks to induce bone loss. The OVX rats were further divided into five subgroups respectively, with Huahong capsule. (1.0,0. 5,0.25 g·kg-1), vehicle (OVX) , diethylstilbestrol for 12 weeks. The concentrations of osteo-caicin, estradiol in serum, deoxypyridinoline in urine, the variation of proximal end of the vertebrae were determined.RESULTS Compared with model group, administration of Huahong capsule decreased serum osteocalcin and urine deoxypyridinoline and increased serum E2 content.. And significant differences were observed in the Huahong capsule treated rats (1.0 g'kg ' and 0. 5 g·kg-1). The Huahong capsule groups increased the thickness and

  7. “补肾健脾”针刺法对绝经后骨质疏松症模型血清碱性磷酸酶水平的影响%Effect of acupuncture method of"invigorating the kidney and strengthening the spleen"on the level of serum alkaline phosphatase in postmenopausal osteoporosis model

    李灿; 黎喜平; 陈久毅; 沈骏; 秧荣昆; 王建


    目的:观察“补肾健脾”针刺法对去卵巢大鼠模型血清中碱性磷酸酶(ALP)含量的影响。方法:将32只雌鼠等分为假手术组、模型组、针刺组及雌激素组。假手术组不摘除卵巢,余3组摘除双侧卵巢,造成绝经后骨质疏松症模型。假手术组及模型组每日同时灌服等量生理盐水,针刺组针刺双侧足三里穴、肾俞穴、脾俞穴、大杼穴及命门穴、关元穴,雌激素组用己烯雌酚溶液灌胃。3个月后取股动脉血,测血清ALP含量。结果:模型组血清碱性磷酸酶含量比假手术组升高(P<0.05);针刺组与雌激素组血清ALP含量较模型组降低(P<0.01)。结论:去卵巢大鼠血清中碱性磷酸酶含量升高,形成高转换骨质疏松症模型,通过雌激素替代和针刺治疗后,血清ALP含量降低,两种治疗均能对抗高转换型骨代谢,为“补肾健脾”治疗绝经后骨质疏松症提供部分临床作用机理。%Objective:To observe the effect of acupuncture method of"invigorating the kidney and strengthening the spleen"on the content of alkaline phosphatase (ALP) in serum of ovariectomized rats.Methods:32 female rats were divided into the sham operation group,the model group,the acupuncture group and the estrogen group.The sham operation group did not remove the ovaries,while bilateral ovaries were removed in the other 3 groups,to made postmenopausal osteoporosis model.The sham operation group and model group were administered with equivalent saline at the same time.The acupuncture group was treated by acupuncture at bilateral Zusanli and Shenyu, Piyu points,Dazhu caveand Mingmen and Guanyuan point,and the estrogen group was treated with intragastric administration of diethylstilbestrol solution.Femoral artery blood was taken after 3 months,and the content of ALP in serum was measured.Results:The serum alkaline phosphatase levels in the model group were higher than those in the sham operation

  8. 米非司酮配伍米索前列醇用于稽留流产疗效研究%Study on the clinical efficacy of mifepristone combined with misoprostol in treatment of missed abortion



    Objective: To explore the clinical efficacy of mifepristone combined with misoprostol in treatment of missed abortion. Methods: 1 960 patients with missed abortion were selected from the hospital, then they were divided into observation group and control group, 980 patients in each group. The patients in observation group were treated with oral administration of mifepristone combined with misoprostol; while the patients in control group were treated with oral administration of diethylstilbestrol. Curettage or intravenous drip of oxytocin were carried out according to uterine size, complete eurettage of uterine cavity was conducted after spontaneous expulsion of the fetus and placen ta, the conditions of the two groups were observed. Results: The success rates of abortion in observation group and control group were 91.84% and 59. 18%, respectively, the success rate of abortion in observation group was significantly higher than that in control group. Conclusion: Oral administration of mifepristone combined with misoprostol can terminate missed abortion safely and effectively,which has the advantages of good clinical efficacy, less amount of blood loss, simplicity and less dose of drugs, and it can reduce the incidence of postoperative incomplete abortion and avoid the occurrence of induced abortion syndrome.%目的:探讨稽留流产应用米非司酮配伍米索前列醇的临床疗效.方法:选择滕州市中心人民医院收治的稽留流产患者1960例,随机分为观察组和对照组,每组各980例.观察组口服米非司酮配伍米索前列醇;对照组口服己烯雌酚.根据子宫大小行钳刮术或静脉滴注缩宫素,待其自然排出胎儿、胎盘后行清宫术,观察两组清宫情况.结果:观察组流产成功率为91.84%,对照组流产成功率为59.18%,观察组流产成功率明显高于对照组.结论:米非司酮配伍米索前列醇口服能安全有效地终止稽留流产,具有疗效好、出血量少,方法简便、用

  9. 固相萃取tipLC法同时测定畜禽肉中5种雌激素的研究%Simultaneous Determination of % Kinds of Estrogens in Meat of Livestock and Poultry by Solid-phase Extraction and-Liquid Chromatography



    [Objective]To research and establish a rapid and sensitive Solid-phase extraction (SPE) high performance liquid chro-matographic method for the determination of 5 estrogens (estradiol, estriol, estrone, ethynylestradiol, and diethylstilbestrol) in meat of livestock and poultry.[Methods]The SPE experimental conditions were optimized and the method of extraction and purification was studied to improve the sensitivity and accuracy. The SPE column was Oasis HLB 6 cc/200 mg. The elutropic liquid was acetoni-trile. Chromatographic column was Eclipse XDB-C18 S μm 4.6 mm x 150 mm, Diode array detector detecting wavelength was 245 run.[Results]Good separation was achieved in the 5 kinds of estrogens and the standard curve showing a direct ratio between the concentration and the peak area were obtained,correlation coefficient r>0.999 3. Detection limits were in the range of 19.2 -24.2 ng/g. The method precision and accuracy were satisfactory with recovery percentages ranging from 79% to 95% and relative standard deviations lower than 5%.[Conclusion]The method is proved to be satisfactory in precision, accuracy and sensitivity and adapt to the daily detection of livestock and poultry meat hormone.%目的 研究建立固相萃取高效液相色谱法同时测定畜禽肉中5种雌激素(雌二醇、雌三醇、雌酮、炔雌醇和己烯雌酚)的快速而准确的方法.方法 优化固相萃取实验条件,研究样品的提取和净化方法,以提高方法的灵敏度和精确度.固相萃取小柱为Oasis HLB 6 cc/200 mg;洗脱液为乙腈;色谱柱为Eclipse XDB- - C18 5 μm 4.6 mm×150 mm柱;二极管阵列检测器,检测波长为210nm.结果 5种雌激素分离效果好,标准曲线线性良好,相关系数r>0.999 3,方法的检出限为19.2~24.2 ng/g,有较好的准确度和精密度,回收率为79% ~95%,相对标准偏差小于5%.结论 该法操作简便,灵敏、准确、无杂质干扰,适合畜禽肉中激素的日常检测.

  10. Effects of Niizhenshuxin formula on the level of E2 in serum, the expression of ER in arterial wall and histomorphology with ovariectomized and high fat rats%女贞舒心方对高脂去卵巢大鼠血清E2水平、动脉管壁ER表达及组织形态学的影响

    钟栩; 王晨; 王安春; 曾俊杰


    Objective To observe effect of Nuzhenshuxin formula (NF) on the level of E2 in serum, the expression of ER in arterial wall and histomorphology with ovariectomized and high fat rats. Methods 60 female rats were divided into 5 groups: normal ( NG), model (MG), Diethylstilbestrol ( DES) and NF high, low dose groups. After the models were established successfully, the rats were fed with high fat diet and drug at the same time. The rat's body weight was recorded once week. 6 weeks later, the content of total cholesterol (TC), tri-glyceride (TG) , low and high density lipoprotein cholesterol (LDL-C, HDL-C) in serum and the level of E2 and expression of ER in arterial wall were detected, while the artery pipe wall change was observed with microscope. Results After fed with NF for 6 weeks, the weight growth of rats were slowed down, the level of TC, TG and LDL-C were declined, the level of HDL-C was increased, the level of E2 and the expression of ER were raised, while the blood vessel endangium was improved, compared with those of model group (P < 0. 05, P < 0. 01). Conclusions NF can increase the level of E2 and adjust the blood fat through displaying plant estrogen type function, it is beneficial to blood vessel protection for menopause women.%目的 通过观察女贞舒心方(NF)对高脂去卵巢大鼠血清雌二醇(E2)水平、动脉管壁雌激素受体(ER)表达及组织形态学的影响,为本方通过调节雌激素水平而发挥心血管保护作用提供科学依据.方法 将成年雌性大鼠60只随机分为5组,分别为正常组(NG)、模型组(MG)、己烯雌酚(DES)组、NF高、低剂量组.除正常组外,切除双侧卵巢,给予高脂饮食,同时给予相应剂量药物灌服,每周测体重,6w后,测定血清胆固醇(TC)、甘油三酯(TG)、高、低密度脂蛋白胆固醇( HDL-C、LDL-C)含量、血清E2水平及主动脉管壁ER的表达,并观察主动脉管壁形态.结果 NF能减缓大鼠体重增长,降低血清TC、TG、LDL-C水

  11. Functional Mechanism of Shao Fu Zhu Yu Wan in Resisting Menalgia No. 4 Hospital of Shijiazhuang, Hebei Province%少腹逐瘀丸抗痛经作用机制研究

    贺克; 刘姣; 李清


    目的:观察少腹逐瘀丸对缩宫素诱发小鼠痛经的镇痛作用及作用机制.方法:取KM小鼠50只,随机分为空白对照组、模型组、丹莪妇康煎膏组、少腹逐瘀丸高剂量组及低剂量组.除空白对照组外,其余各组小鼠灌胃给药同时灌服乙烯雌酚,连续7天.末次给药30 min后,除空白对照组外,其余各组小鼠腹腔注射缩宫素,观察小鼠离体子宫平滑肌收缩变化,小鼠30 min内扭体反应次数,并检测小鼠血浆血栓烷B2(TXB2)、6-酮-前列腺素F1α(6-Keto-PGF1α)及血清超氧化物歧化酶(SOD)、丙二醛(MDA)含量.结果:少腹逐瘀丸能够明显抑制缩宫素诱导小鼠离体子宫平滑肌收缩(P<0.05),使痛经小鼠扭体反应次数有降低趋势,但差异无显著性(P>0.05),并能明显降低小鼠血浆TXB2含量(P<0.05),明显升高血浆6-Keto-PGF1α含量(P<0.05),显著降低血清MDA含量(P<0.01),明显升高血清SOD含量(P<0.05).结论:少腹逐瘀丸对缩宫素诱导小鼠痛经模型具有较好治疗作用,其作用可能与调节TXB2/6-Keto-PGF1α比值、抗氧化有关.%Objective: to observe the analgesic effect and the functional mechanism of Shao Fu Zhu Yu Wan on oxytocin - induced menalgia of mice. Method: 50 KM mice were randomly divided into control group, model group, soft extract group of Dan'e Fukang, high dose and low dose groups of Shao Fu Zhu Yu Wan, all of which, except the first group, were given intragastric administration and diethylstilbestrol for 7 days successively, then in-traperitoneal injection with oxytocin 30 minutes after the last administration, for the purpose to observe the contraction changes of the uterus smooth muscle in vitro, frequency of writhing response in 30 minutes and the following contents: TXB2 , 6 - Keto - PGF1, SOD and MDA. Results: Shao Fu Zhu Yu Wan could inhibit significantly the muscle contraction of the uterus in vitro induced by oxytocin ( P 0. 05 ), significantly decrease TXB2( P <0. 05

  12. Study of expression of neuropathic nitric oxide synthase, endothelial nitric oxide synthase and inducible nitric oxide synthase in penile tissue of erectile dysfunction rat models with prolactinoma%泌乳素瘤性阴茎勃起功能障碍大鼠阴茎各亚型一氧化氮合酶表达的变化

    翁博文; 祝海; 侯四川; 徐珞; 栾晓; 綦海燕; 刘之俊; 王伟民; 刘伟


    目的 探讨泌乳素瘤性勃起功能障碍的发病机制.方法 雄性Wistar大鼠腹腔内注射乙烯雌酚(DES)建立泌乳素瘤模型.8周通过阿扑吗啡(APO)实验筛选,建立泌乳素瘤性阴茎勃起功能障碍(P-ED)模型,应用免疫组织化学方法测定大鼠阴茎组织神经型一氧化氮合酶(nNOS)、内皮型一氧化氮合酶(eNOS)和诱导型一氧化氮合酶(iNOS)表达水平的变化.结果 DES注射8周后,垂体泌乳素瘤成模率为100%,P-ED成模率为75%.与对照组比较,注射DES 8周时大鼠垂体质量明显增加;血清泌乳素(PRL)水平升高而游离睾酮(FT)水平降低;阴茎勃起次数显著减少.阴茎组织nNOS、eNOS表达降低而iNOS表达增加.结论 P-ED大鼠模型阴茎组织nNOS、eNOS表达减少而iNOS表达增加.%Objective To investigate the mechanism of erectile dysfunction of prolactinoma.Methods Male Wistar rats were treated with diethylstilbestrol (DES) by peritoneal injection to establish the rat model of prolactinoma.After 8 weeks,the model rats were injected with apomorphine to screening ED rats models with DES-induced prolactinoma (P-ED).The changes of expression of neuropathic nitric oxide synthase (nNOS),endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS) in penis were detected with immunohistochemistry.Results By 8 weeks,100% pituitary glands of DES group developed prolactinomas and 75% DES-induced prolactinoma rats developed ED models.The weight of pituitary gland was dramatic increased.The Prolactin (PRL) level of P-ED group was significantly higher and FT level was lower than the control group.Erectile rate of DES group was significant lower than the control group after 8 weeks of DES injection.The expression of nNOS and eNOS in penis of P-ED group was significantly lower than the control group.However,the expression of iNOS in penis of P-ED group was significantly higher than the control group.Conclusion The expression of nNOS and e

  13. Fate and removal of typical pharmaceuticals and personal care products by three different treatment processes

    He, Yu-jie; Chen, Wei; Zheng, Xiao-ying, E-mail:; Wang, Xing-nan; Huang, Xi


    The presence and distribution of typical of pharmaceuticals and personal care products (PPCPs), which comprise two types of polycyclic musks (PCMs) including Galaxolide (HHCB) and Tonalide (AHTN) as well as six types of estrogens containing estrone (E1), 17β-estradiol (E2), estriol (E3), 17α-ethynylestradiol (EE2), diethylstilbestrol (DES), and bisphenol A (BPA), were investigated at two wastewater treatment plants (WWTPs) in Jiangsu, China. Only raw wastewater was treated in WWTP A while WWTP B was serving an urban-industrialized area. In the influent, the concentrations of EE2 (2193–4437 ng L{sup −1}), E2 (1126–1170 ng L{sup −1}), and DES (268–421 ng L{sup −1}) were generally higher than the previously reported values, whereas the concentrations of HHCB (306–316 ng L{sup −1}), E1 (29–129 ng L{sup −1}), E3 (53 ng L{sup −1}), and BPA (26–176 ng L{sup −1}) were much lower than those reported in other previous studies. In addition, AHTN was not detected in either WWTP and E3 was not found in WWTP B. The detected processes including anaerobic/oxic process (A/O), combined orbal oxidation ditch process (C-orbal OD) and anaerobic/anoxic/anoxic/oxic membrane biological reactor (A/A/A/O-MBR) showed higher removal efficiencies for HHCB (67–71%) and EE2 (87%) than those in other previous studies. Besides, the total hydraulic retention time (HRT) ranged between 6.7 and 20.0 h, sludge retention time (SRT) ranged between 8 and 23 d, and water temperature ranged from 24.8 to 28.2 °C. The removal efficiencies for estrogens in biological processes were related to the following factors: the level of hydrophobic estrogens, the type of removal process (C-orbal OD was consistently less efficient in removing estrogens than A/O and A/A/A/O-MBR), and a high SRT or HRT (A/A/A/O-MBR with higher SRT and HRT showed higher and more stable removal of hydrophobic estrogens). - Highlights: ► We investigated 8 kinds of PPCPs in each unit at 2 WWTPs with different

  14. 双酚A对成年雄性小鼠食物利用率的影响%Effect of bisphenol-A on food utilization rate in adult male mice

    赵海军; 马丽娟; 朱玉莲; 陈敏华; 陈哲科


    目的 探讨双酚A对成年雄性小鼠食物利用率的影响.方法 100只成年雄性ICR小鼠随机分为阴性对照组、己烯雌酚对照组、双酚A2、20、100 mg/(kg·d)组,各组每天给予相应剂量受试物灌胃2周,之后再观察6周.实验期间每周称量各组动物体质量1次,每天称量各组动物摄取生长繁殖颗粒饲料的重量.结果给予双酚A8周后,各组动物体质量差异无统计学意义(P>0.05);给予双酚A的第2周,2,20,100mg/(kg·d)双酚A组动物的食物利用率与阴性对照组比较明显升高(P<0.01);第4周,20,100mg/(kg·d)双酚A组动物的食物利用率与阴性对照组比较明显升高(P<0.05);第6周,100mg/(kg·d)双酚A组动物的食物利用率与阴性对照组比较明显升高(P<0.05);第8周,所有双酚A组动物的食物利用率与阴性对照组比较差异无统计学意义(P>0.05).结论 双酚A对于成年雄性ICR小鼠的食物利用率有增高作用.%Objective To investigate the effect of bisphenol-A on food utilization rate in adult male mice. Methods A total of 100 adult male ICR mice were randomly divided into groups of negative control. 20 mg/kg diethylstilbestrol control, and 2, 20 and 100 mg/kg bisphenol-A treatment, respectively. Mice of each group were treated with relative material once per day by intragastric administration for two weeks and observed for six weeks after treatment. The weight of mice was observed by weekly measurement and the weight of food intake was taken by daily recording. Results There was no significantly difference in body weight among all groups in the sixth week after bisphenol-A treatment. Food utilization rate was higher in the groups of 2, 20 and 100 mg/kg bisphenol-A treatment than in negative control group with two weeks of bisphenol-A treatment (P<0. 01). Food utilization rate was still higher in the groups of 20 and 100 mg/kg bisphenol-A treatment than in negative control group in the second week after bisphenol

  15. On the rumors about the silent spring: review of the scientific evidence linking occupational and environmental pesticide exposure to endocrine disruption health effects Rumores de uma primavera silenciosa: uma revisão das evidências científicas sobre a associação entre exposição ocupacional e ambiental a pesticidas e distúrbios endócrinos

    Pierluigi Cocco


    Full Text Available Occupational exposure to some pesticides, and particularly DBCP and chlordecone, may adversely affect male fertility. However, apart from the therapeutic use of diethylstilbestrol, the threat to human reproduction posed by "endocrine disrupting" environmental contaminants has not been supported by epidemiological evidence thus far. As it concerns other endocrine effects described in experimental animals, only thyroid inhibition following occupational exposure to amitrole and mancozeb has been confirmed in humans. Cancer of the breast, endometrium, ovary, prostate, testis, and thyroid are hormone-dependent, which fostered research on the potential risk associated with occupational and environmental exposure to the so-called endocrine-disrupting pesticides. The most recent studies have ruled out the hypothesis of DDT derivatives as responsible for excess risks of cancer of the reproductive organs. Still, we cannot exclude a role for high level exposure to o,p'-DDE, particularly in post-menopausal ER+ breast cancer. On the other hand, other organochlorine pesticides and triazine herbicides require further investigation for a possible etiologic role in some hormone-dependent cancers.A exposição ocupacional a determinados pesticidas, particularmente ao DBCP e à clordecona, pode ter efeitos adversos sobre a fertilidade masculina. Entretanto, com exceção do uso terapêutico do dietil-estilbestrol, a ameaça à reprodução humana através da "desregulação endócrina" por contaminantes ambientais ainda não foi comprovada através de evidências epidemiológicas. A questão diz respeito a outros efeitos endócrinos descritos em animais experimentais, e apenas a inibição tireóide foi confirmada em seres humanos, após exposição ocupacional a amitrole e mancozeb. O fato de serem hormônio-dependentes os cânceres de mama, endométrio, ovário, próstata, testículos e tireóide motivou pesquisas sobre o risco potencial associado à exposi

  16. Testicular dysgenesis syndrome and the estrogen hypothesis: a quantitative meta-analysis A síndrome da disgenesia testicular e a hipótese do estrogênio: uma meta-análise quantitativa

    Olwenn Martin


    Full Text Available Male reproductive tract abnormalities such as hypospadias and cryptorchidism, and testicular cancer have been proposed to comprise a common syndrome together with impaired spermatogenesis with a common etiology resulting from the disruption of gonadal development during fetal life, the testicular dysgenesis syndrome (TDS. The only quantitative summary estimate of the link between prenatal exposure to estrogenic agents and testicular cancer was published over 10 years ago; other reviews of the link between estrogenic compounds, other than the potent pharmaceutical estrogen diethylstilbestrol (DES, and TDS end points have remained inconclusive. We conducted a quantitative meta-analysis of the association between the end points related to TDS and prenatal exposure to estrogenic agents. Inclusion in this analysis was based on mechanistic criteria, and the plausibility of an estrogen receptor (ER-α-mediated mode of action was specifically explored. Eight studies were included, investigating the etiology of hypospadias and/or cryptorchidism that had not been identified in previous systematic reviews. Four additional studies of pharmaceutical estrogens yielded a statistically significant updated summary estimate for testicular cancer. Results of the subset analyses point to the existence of unidentified sources of heterogeneity between studies or within the study population.Sugeriu-se que anomalias do trato reprodutivo masculino como hipospádia e criptorquidismo, assim como o câncer de testículo, componham uma síndrome comum com diminuição da espermatogênese, e de etiologia comum, a interrupção do desenvolvimento gonadal na fase fetal, a síndrome de disgenesia testicular (SDT. O único levantamento quantitativo da relação entre exposição pré-natal a agentes estrogênicos e câncer de testículo data de mais de dez anos; outras revisões da relação entre compostos estrogênicos diferentes do potente estrogênio sint

  17. Explanation of the mechanism of carcinogenesis and syntheses of anticancer agents with high selectivity


    In 1979, the mechanism of chemical carcinogenesis, a challenging and difficult scientific problem pending for a number of years, was explained by Dai Qianhuan. The mechanism named di-region theory predicted that a carcinogen always metabolizes to form a special bi-functional alkylating agent. This agent induces cross-linkages between the complementary base pairs in DNA and switches on initial mutageneses in genomes including point and frameshift mutations. This, in turn, induces further deep mutageneses including the production of various chimeric chromosomes, deletions and other aberrations found in genomes. In the end this initiates carcinogenesis of the whole cell through the reverse transcription mechanism after a lengthy incubation period. Recently, this laboratory has verified that physical carcinogenesis, including the oncogenesis induced by radiation and asbestos as well as the carcinogenesis induced by endogenous factors such as estrogen or diethylstilbestrol switch on carcinogenesis by inducing the formation of cross-linkages between the complementary base pairs in DNA. Di-region theory has now been supported by many experimental observations such as mutational spectra of various carcinogens. The potential for carcinogenesis, teratogenesis, sterility and mutagenesis lumped together as genetic toxicity appears to originate almost uniformly from the cross-linking between complementary bases, i.e. malignant cross-linking, which is in accordance with di-region theory. Other forms of cross-linking between non-complementary bases, benign cross-linkings, show bi-functional alkylation anticancer activity but lack genetic toxicity. The predictable design and synthesis of a high selectivity anticancer agent with high efficacy and low genetic toxicity, a goal long pursued in cancer chemotherapy, have been realized for the first time in this laboratory by inhibiting malignant and heightening benign cross-linking using the principles of di-region theory. A series of

  18. Effects of Xiaoyu Tongluo San on breast morphology and endocrine in rats with hyperplasia of mammary glands%消瘀通络散对乳腺增生大鼠乳腺组织形态与内分泌的影响

    张拴成; 王丽娜; 肖红玲; 佘延芬; 边文静; 郭帅; 杨继军


    目的:观察自拟消瘀通络散对乳腺增生大鼠乳头直径、高度,血清E2/P、PRL、T水平以及乳腺组织形态的影响,探讨其治疗乳腺增生的作用机理.方法:将40只Wistar大鼠随机分为正常组、模型组、中药组和西药组4组.采用己烯雌酚联合黄体酮肌注的方法进行造模.中药组给予自拟消瘀通络散50mg/d,西药组予三苯氧胺1.8mg/kg,均灌胃治疗,1次/d,共30d.于治疗前后测量各组大鼠第2对乳头直径、高度,测定血清E2、P、PRL、T水平,计算E2/P数值,取大鼠第2对乳房常规HE染色,光镜下观察组织形态表现.结果:治疗后乳头直径、高度比较,中药组和西药组均低于模型组(P<0.05);2组之间比较,中药组低于西药组(P<0.05).血清各激素水平比较,中药组和西药组血清E2/P、PRL、T含量均低于模型组(P<0.05);中药组与西药组相比无显著性差异(P>0.05).病理学比较,中药组和西药组大鼠乳腺组织病理表现相似,与模型组比较改善明显,与正常组相近.结论:自拟消瘀通络散具有缩小模型大鼠乳头直径、高度,改善内分泌紊乱状态和乳腺组织病理形态的作用.%Objective: To observe the effect of Xiaoyu tongluo san on the second pair of breast nipple diameter, height and serum E2/P ratio, PRL, T levels ^breast tissue of the Mammary Gland Hyperplasia rats model, discuss the therapeutic mechanism of HMG. Methods; 40 Wistar rats were divided into 4 groups randomly:normal group, model group, TCM group and western medicine group. Combined diethylstilbestrol with progesterone by intramuscular injection to copy breast Hyperplasia rats model. TCM group were treated with self-Xiaoyu tongluo san 50mg/d, while western medicine group were treated with TAM 1. 8mg/kg,gavage was both applied, 1 time/d, 30 days totally. Detected the second pair of breast nipple diameter ,height and serum E2/P ratio, PRL, T levels before and after treatment,calculate E2/P ratio, The

  19. 雷洛昔芬对放疗去势大鼠骨丢失的保护作用%Protective effect of raloxifene in preventing bone loss in rats following radiotherapy induced gonadectomy

    陈静; 朱芳; 李贵玲


    目的:观察雷洛昔芬(选择性雌激素受体调节剂)对放疗去势大鼠的骨代谢生化指标、骨密度及骨形态学的影响,探讨其对放疗所致卵巢早衰引起骨丢失的保护作用.方法:选用3月龄雌性Wistar大鼠40只,随机分为4组:假放疗组、放疗组、放疗+雷洛昔芬组、放疗+己烯雌酚组,每组10只,16周后,腹主动脉取血,检测血清雌二醇、卵泡刺激素、骨钙素及碱性磷酸酶的水平;处死大鼠,取大鼠右侧股骨,测定各组骨密度;同时取左侧股骨制作骨切片行HE染色,观察骨形态学变化.结果:放疗组及放疗+雷洛昔芬组的雌二醇水平明显低于假放疗组及放疗+己烯雌酚组(P<0.05);而两组的卵泡刺激素水平明显高于假放疗组及放疗+己烯雌酚组(P<0.05);放疗组骨钙素和碱性磷酸酶明显高于其他3组,而另3组无明显差异(P>0.05);放疗组大鼠骨密度显著低于其他3组,其骨小梁稀疏,部分断裂,而另3组骨小梁致密,无明显断裂.结论:大鼠卵巢放疗可致去势,放疗去势后大鼠有骨质丢失发生,而选择性雌激素受体调节剂雷洛昔芬对放疗去势后大鼠的骨丢失有保护作用.%OBJECTIVE To investigate the protective effect of the selective estrogen receptor modulators (SERMs) raloxifene in preventing hone loss secondary to radiotherapy induced premature ovarian failure(POF). This was studied by observing the effects of Raloxifene on biochemical markers of bone metabolism, bone morphology and bone mineral density of rats following radiotherapy induced gonadectomy. METHODS 40 female Wistar rats aged 3 months were randomly divided into 4 groups (10 rats in each group): Group A(sham radiotherapy), Group B(radiotherapy),Group C (radiotherapy and raloxifene) ,Group D(radiotherapy and diethylstilbestrol). All rats were killed 16 weeks later. Then serum estradiol (E2)、 follicle stimulating hormone(FSH),bone gla protein(BGP) and alkaline phosphatase

  20. Different ingredients of cell culture medium influence multi-differentiation of bone marrow mesenchymal stem cells%不同因子影响骨髓间充质干细胞的多向分化

    栗扬阳; 赵庆华


    medium on oriented differentiation of bone marrow mesenchymal stem cel s. METHODS: A computer-based online retrieval of PubMed and Wanfang databases was performed to search papers published between January 1998 and April 2012 using the key words “bone marrow mesenchymal stem cel s, cel culture medium, differentiation” in English and Chinese, respectively. Papers regarding effects of different ingredients of culture medium on osteogenic, chondrogenic and adipogenic differentiation of bone marrow mesenchymal stem cel s were col ected. Papers with repetitive contents were excluded. RESULTS AND CONCLUSION: A total of 184 papers were initial y retrieved. According to inclusion and exclusion criteria, 30 of them were suitable for final analysis. General y speaking, dexamethasone, transforming growth factor, vitamin C, vitamin D3, β-sodium glycerophosphate, and diethylstilbestrol are the main ingredients of cel culture medium to induce osteogenic differentiation of bone marrow mesenchymal stem cel s; dexamethasone, transforming growth factor, vitamin C, insulin-like growth factor and fibroblast growth factor are the main ingredients of cel culture medium to induce chondrogenic differentiation of bone marrow mesenchymal stem cel s; dexamethasone, 3-isobutyl-1-methylxanthine, insulin and indometacin are the main ingredients of cel culture medium to induce adipogenic differentiation of bone marrow mesenchymal stem cel s. Nevertheless, the mechanisms of action and adverse events of some ingredients are poorly understood and need further investigation. In addition, bone marrow mesenchymal stem cel s are at low level in the bone marrow and different isolation methods wil lead to different cel proportions. Therefore, a method of isolating high proportion of cel s should be developed.

  1. 同位素稀释-固相萃取-超快速液相色谱/串联质谱联用法测定鳝鱼中9种雌激素%Simultaneous determination of nine estrogens in eel by ultrafast liquid chromatography-tandem mass spectrometry with isotope dilution technique and solid-phase extraction

    陈晓红; 姚珊珊; 李小平; 赵永纲; 金米聪


    Objective Developing a rapid and sensitive analytical method based on ultrafast liquid chromatography-tandem mass spectrometry ( UFLC-MS/MS) with solid-phase extraction ( SPE) for the simultaneous determination of nine estrogens (dienestrol, diethylstilbestrol, estrone, hexestrol, 17-alpha-estradiol, 17-beta-estradiol, estriol, 17α-ethinylestradiol and estradiol valerate) in eel. Methods After the sample was extracted by acetonitrile and cleaned by Waters Oasis" HLB solid-phase extraction cartridge, the UFLC separation was performed on a Shim-pack XR-ODS II column ( 100mm × 2. 0mm, 2. 2μm) with a linear gradient elution program of methanol solution containing 0.04% ammonia (v/v) and 0.04% ammonia aqueous solution ( v/v) as the mobile phase. Electrospray ionization was applied and operated in the negative multiple reaction monitoring (MRM) mode. The quantitation was used by isotope internal standard dilution technique. Results The results showed that the limits of quantitation (LOQs, S/N(10) were in the range of 0. 07-0. 4μg/kg, the calibration curves were in good linearities for the nine analytes in the range of 0. 5-50. 0μg/L with the correlative coefficients ( r2 ) more than 0. 998 , the recoveries were between 81. 0% and 110. 0% with the relative standard deviations (RSDs) of 1.92%-8. 24%. Additional, the mass spectra characterization of the nine estrogens was discussed and the fragmentation pathways were speculated. Conclusion The developed method is rapid, sensitive, specific and reproducible, and adapts not only to the simultaneous determination of the nine trace estrogens including the epimer of 17-alpha-estradiol and 17-beta-estradiol but also to the identified detection in other fish tissues.%目的 建立一种快速、灵敏的鳝鱼中己二烯雌酚、己烯雌酚、雌酮、己烷雌酚、17α-雌二醇、17β-雌二醇、雌三醇、17α-炔雌醇和戊酸雌二醇等9种雌激素残留的固相萃取超快速液相色谱-串联质谱(UFLC-MS/MS

  2. Effects of genistein on bone mineralization and osteoporosis in ovariectomized rats%染料木黄酮对去势大鼠骨骼矿化及骨质疏松的影响

    张月红; 金宏; 许志勤; 南文考; 王先远; 薛长勇; 高兰兴


    BACKGROUND: Genistein is the main component of phytoestrogen soy isoflavone and its structure is similar to estrogen,which suggests that it might prevent or delay osteoporosis. Research on the effects of genistein on bone mineralization and calcium(Ca), phosphor(P), zinc(Zn) and magnesium (Mg) in ovariectomized rats are rare.OBJECTIVE: To investigate the effects of genistein on bone mineralization and Ca,P,Zn and Mg in ovariectomized rats to provide a theoretical gist for the prevention of osteoporosis by genistein.DESIGN: A controlled experiment based on experimental animals.SETTING: Department of Nutrition,General Hospital of Chinese PLA.MATERIALS: The study was conducted in the Institute of Hygiene and Environmental Medicine,Academy of Military Medical Sciences of PLA between February and June 2003. Ten-week old female Wistar rats [certification number: (military medical animal): D98014] with a body mass of(170±20) g were selected.INTERVENTIONS: Experimental animals were fed with normal feeding for 6 weeks and then the feeding was changed to AIN-93 compound. Animals were then randomly divided into ovariectomized group (n=40) and sham-operation group(n=7) based on bodyweight after 5 days. Ovariectomized group received ovariectomy and sham-operation group only received abdominal incision. After 5 days of recovery,the ovariectomized group was further randomly divided into 5 subgroups with 8 rats each including ovariectomized control subgroup,estrogen subgroup [diethylstilbestrol 20 μg/(kg · d)],genistein Ⅰ,Ⅱ,or Ⅲ subgroup[dose of 25,50 or 100 mg/(kg · d)]. After 3months of feeding,6 rats were randomly selected from each group for the detection of bone density and corresponding bone hismorphometric indicators.MAIN OUTCOME MEASURES: Bone density,corresponding parameters of bone mineralization,Ca,P,Zn and Mg contents in bone of mice in each group RESULTS: After ovariectomy,femoral bone density decreased [(0. 247± 0.007) g/cm2],mean osteoid width increased

  3. Determination of Estrogens in Water Samples From Wastewater Treatment Plant Using Ultra-Performance Liquid Chromatography-Electrospray Tandem Mass Spectrometry Method%超高效液相色谱-串联质谱法测定污水处理厂水样中的雌激素

    谭丽超; 葛峰; 单正军; 王懿


    An instant estrogen screening and determination method, using the HLB solid-phase extraction and ultra-performance liquid chromatography-electrospray tandem mass spectrometry technology ( UPLC-MS/MS) in a multiple reaction monitoring ( MRM ) mode, was established for determination of seven estrogens ( estradiol valerate, ethynylestradiol, estriol, estradiol, hexestrol, estrone and diethylstilbestrol) in the influent and final effluent of a wastewater treatment plant. Conditions for performances of solid-phase extraction ( selection of extraction cartridge, eluate and its volume used for elution, pH adjustment of the water sample, and rinsing solution) and for UPLC -MS/MS analysis (mobile phase and gradient, capillary voltage, cone voltage, RF lens voltage, collision energy, ion source temperature, desolvation gas flow and cone gas flow) were optimized. In the case that all the seven estrogens were in the linear range of 1 -100μg ? L-1, in mass concentration, all the determination coefficients ( r2 ) of the regression equation were over 0. 997 2, and the detection limit of the method ranged from 4. 40 to 10. 27 ng ? L~' for estrogens. When the mass concentration of estrogens was raised to the range of 20 -100 ng ? L-1, the recovery rates of the target substances in the influent and effluent water samples were in the range from 70. 3% to 95. 1% and from 72. 6% to 96. 7% , separately, with relative standard deviation (RSD) below 10. 4% and 9. 5% , separately. This method was applied to determine estrogens in influents and effluents of four major integrated wastewater treatment plants in Nanjing. It was found that concentrations of estradiol valerate, estriol and estrone reached as high as dozens and even hundred of ng ? L-1 in the influents and several or dozens in the final effluents.%采用HLB固相萃取、超高效液相色谱-串联质谱(UPLC - MS/MS)法,在多反应监测(MRM)模式下建立污水处理厂进、出水口水样中7种雌激素(戊酸雌二醇


    迟晓星; 张涛; 陈容; 李蓉


    能是引起PCOS发生的因素之一.Gen可有效调节PCOS高雄血症大鼠血清性激素水平,对大鼠卵巢组织中FSHR蛋白及LHR蛋白均有调节作用,提示其对PCOS可起到治疗作用.%Objective To study the therapeutic effect of genistein (GEN) on serum sex hormone and luteinizing hormone receptor (FSHR) protein, luteinizing hormone receptor (LHR) protein of ovary in female rats with polycystic ovary syndrome (PCOS) and hyperandrogenism. Method The insulin (INS) combined with human chorionic gonadotropin (HCG) molding method was used to establish PCOS animal model. Sixty rats were divided into control group, model group, low, middle, high dose Gen groups (L-G, M-G, H-G) given Gen 5, 10, 20 mg/kg bw respectively and estrogen group (EG), given diethylstilbestrol 0.5 mg/kg bw, All compounds were administered i.g on 15 consecutive days and the estrous cycle was observed for 10 d. The levels of progesterone (P), testosterone (T), sex hormone binding globulin (SHBG), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in serum and FSHR protein, LHR protein of ovary were examined. Results Compared with the model group, the level of P increased significantly in M-G, H-G, and EG groups (P<0.01). The level of T decreased significantly in M-G, H-G and EG (P < 0.01). The level of LH decreased significantly in all dose groups (P<0.01). The level of FSH increased significantly in H-G and EG (P<0.01). The ratio of LH/FSH decreased significantly in M-G, H-G and EG (P< 0.01). Immunohistochemical results showed that compared with the model group, the expression of FSHR protein in rats' ovarian granulosa cells decreased after giving Gen (10,20 mg/kg bw·d). The average positive area and average density decreased. LHR protein expression increased. The average positive area and average density increased (P<0.01). Conclusion High level of LH in serum increased T concentration and hyperandrogenism may be the reasons for PCOS. Concentration of sex hormones


    陈小波; 程飚; 刘宏伟; 孙同柱; 付小兵


    ESCs were isolated from normal human foreskin and cultured. The second generation of hESCs were identified with flow cytometry after being marked with integrin β1, cytokeratin 19 (CK19), CK14, and CK10 antigens.hESCs of 2 × 106 cell density were cultured with ESCs special medium supplemented with 0.1 nmol/L Diethylstilbestrol in group A (estrogen group), with ESCs special medium supplemented with 10 nmol/L Raloxifene hydrochloride in group B (ER blocking agent group), and with ESCs special medium in group C (control group), respectively. The 100 μm “scratch” wounds were created by scraping confluent hESCs plated on Petri dishes with a sterile pipette tip in vitro. The migrating cells from the wound edge were quantified at 24, 48, and 72 hours after incubation. The rates of wound healing were calculated by SigmaScan Pro 5.0 software at 72 hours. The proliferating effect of estrogen on hESCs was determined with MTT method at 24, 48, 72, 96,and 120 hours. Results Cultured primary hESCs could adhere to the wall showing ovoid in shape and grew into colonies.Flow cytometry showed the positive results for integrin β, CK19, and CK14 (with positive rate of 96.63%, 95.47%, and 94.27%,respectively) and the negative result for CK10 (with positive rate of 1.32%). In group A, the number of cells crossing the wound edge was more than those of group B and group C at 24, 48, and 72 hours. The rates of wound healing were 69.00% ± 0.05%in group A, 35.00% ± 0.05% in group B, and 48.00% ± 0.06% in group C at 72 hours, showing significant differences among groups (P < 0.05). The proliferating speed of hESCs was significantly higher in group A than in groups B and C (P < 0.01), and significantly higher in group C than in group B (P < 0.01) at 24, 48, 72, 96, and 120 hours. Conclusion The estrogen can promote the proliferation and migration of hESCs in vitro. It may be involved in many biological activity of skin.

  6. Study on Rapid Detection Technology on Banned Veterinary Drug Residues in Livestock Products%畜禽产品中重要违禁兽药残留快检技术研究

    何方洋; 郗日沫; 万宇平; 罗晓琴; 冯才伟


    With the rapid development of socio-economic and the increasing living standards, people paid more attention on food security,including the increasingly importance on security of livestock products. This study mainly research on the development on rapid detection kit including hormones, nitroimidazoles and olaquindox residual in livestock products, and colloidal gold test strip technology including β-lactam and β-lactamase in dairy products. We had developed seven detection kit concluding diethylstilbestrol, hydrocortis one,dexamethasone,estradiol,19-nortestosterone, nitroimidazoles,olaquindox metabolites,and 5 test strip products containing hydrocortisone ,trenbolone,dexamethasone,β-lactam antibiotics,β-lactamase.The R&D of ELISA kits base on the principle of the combination of specific immune response between antigen and antibody,and amplification reaction of enzyme-linked response,characterized by specific targeted on various drug molecules in the test samples.The Sensitivity reached 0.05~0.5 μg/kg by enzyme-linked amplification,achieving the detection limits of analytical instruments as high performance liquid chromatography, liquid chromatography - mass spectrometry and other sophisticated analytical instruments.It could realize qualitative and semi-quantitative detection in sample residues; shorten detection time from the previous instrument detects one day or several days to no more than two hours,greatly improving the detection efficiency. It can synchronous detect tens to hundreds of samples rapidly, meeting the detection needs of extensive and comprehensive inspection on food safety; low cost detection, costing only 1/5~1/2 the Instrument testing, can be widely used in the monitoring and testing work of the regulatory functions of the government grassroots, small and medium enterprises without large detection equipment. The development of colloidal gold test strip is a novel gold standard plastic immune technique ,based on colloidal gold labeling

  7. 雌激素对运动防治骨质疏松效果的影响%Effects of estrogen on outcomes of exercise therapy for osteoporosis

    陈柏龄; 黎艺强; 谢登辉; 廖威明; 李佛保


    Objective To study the effects of estrogen on outcomes of exercise therapy for osteoporosis. Methods Forty-eight 5-month-old female SD rats were randomly divided into following groups:sham operated rats (Sham group), sham operated rats on exercise therapy (Sham+run group), ovariectomy rats (Ovx group), Ovx rats on exercise therapy (Ovx+run), and Ovx rats on exercise therapy and 1/4-dose (Ovx+run+e1 group) or full-dose (Ovx+run+e2 group) estrogen. For the last two groups, subcutaneous injection of diethylstilbestrol was started one week after oophorectomy at a dose of 0.025 mg/kg once every 4 days (Ovx+run+e1 group) or once daily (Ovx+run+e2 group) and lasted for 12 weeks. Exercise therapy,designed as in-cage running, was also started one week after operation in the Sham+run, Ovx+run, Ovx+run+e1 and Ovx+run+e2 groups and lasted for 12 weeks. On week 13, bone mineral density (BMD) and bone biomechanical parameters of distal femur and bone histomorphometry of proximal tibia were measured.Results (1)Ovx group had the lowest B MD [ (0.10 ± 0.01 ) g/cm2] compared statistically with the other groups (all P<0.05). Ovx+run+e2 [ (0.14±0.02) g/cm2] and Sham+run [ (0.13±0.02) g/cm2] groups had BMD that was comparable to each other but significantly higher than the other groups (all P<0.05). No statistical difference in BMD could be found among Sham [ (0.11 ±0.01) g/cm2], Ovx+run [ (0.12±0.01)g/cm2] and Ovx+run+e1 [(0.12±0.01 ) g/cm2] groups (all P>0.05). (2)The trabecular thickness, percentage area of trabeculae and trabecular bone mass in Sham group were comparable to those in Ovx+run+e2 and Sham+run groups but were higher than those in Ovx+run+el, Ovx+run and Ovx groups (all P<0.05). Sham,Ovx+run+e2 and Sham+run groups had trabecular separation that was comparable among each other but less than those in Ovx+run+e1, Ovx+run and Ovx groups (all P<0.05). The percentage area of trabeculae and the trabecular bone mass increased while trabecular separation

  8. 维生素K2及其配伍制剂抗实验性骨质疏松症的筛选研究%Anti-osteoporotic screening study of vitamin K2 and its compatibility of medicines

    陈鹏; 何波; 杨仁华; 陆义芹; 沈志强


    ; vitamin K2 (80 g/kg) + calcium citrate (500 mg/kg), sample 5: vitamin K2 (50 g/kg) + vitamin D3 (4 g/kg), sample 6: vitamin K2 (80 g/kg) + vitamin D3 (4 g/kg), sample 7: vitamin K2 (50 g/kg) + vitamin E (40 mg/kg), sample 8: vitamin K2 (80 g/kg ) + vitamin E (40 mg/kg), sample 9: vitamin K2 (50 g/kg) + vitamin E (40 mg/kg) + vitamin D3 (4 g/kg), sample 10: vitamin K2 (80 g/kg) + vitamin E (40 mg/kg) +- vitamin D3 (4 g/kg), sample 11: vitamin K2 (50 g/kg ) + vitamin E (40 mg/kg) + calcium citrate (500 mg/kg), sample 12: vitamin K2 (80 g/kg ) + vitamin E (40 mg/kg) + calcium citrate (500 mg/kg), sample 13: vitamin K2 (50 g/kg) + vitamin D3 (4 g/kg) + calcium citrate (500 mg/ kg), and sample 14: vitamin K2 (80 g/kg) + vitamin D3 (4 g/kg) + calcium citrate (500 mg/kg). Female Sprague-dawly rats were randomly divided into normal control group, sham operation group, and ovariectomized (OVX) group. Rat osteoporosis model was established using OVX. Rats in OVX group were then randomly divided into model group, 30μg/kg diethylstilbestrol positive control group, and the above 14 test sample groups, respectively. Rats in 14 test sample groups were given a 1ml/100g administration of the samples intragastrically once a day, lasting for 60 days. Normal saline was used in normal control group, and peanut oil was used in sham and model groups, respectively. The detailed grouping was shown in Table 1. Bone mineral density (BMD) and bone mineral content (BMC) were examined by dual-energy X-ray absorptiometry analysis, and bone calcium by atomic absorption, respectively. Results Effect on BMD and BMC : as compared with that in model group, test sample 13 and 14 could significantly improve BMD and BMC of the femur and the vertebrae ( P < 0. 01). The effect of test sample 14 was stronger than that of test sample 13 (P < 0. 05). Test sample 1 and test samples 2, 4, 6, 8, 10, and 12 ( combined with the same dose of test sample 1) could increase BMD and BMC of the femur and the vertebrae