Characterization of estrogen-degrading bacteria isolated from an artificial sandy aquifer with ultrafiltered secondary effluent as the medium

This study investigated the aerobic and anoxic biodegradation of four estrogens [estrone (E1), estradiol (E2), estriol (E3), and the synthetic 17α-ethinylestradiol (EE2)] in microcosms constructed with marine sand and ulftrafiltered (UF) secondary effluent. Three estrogen-degrading bacteria, LHJ1, LHJ3, and CYH, were isolated. Based on gram-stain morphology and 16S rRNA sequence homology, LHJ1 and LHJ3 belong to the genus Acinetobacter and Agromyces, respectively; CYH matched to 95% with the genus Sphingomonas. Aerobically LHJ3 degrades E3, CYH degrades E1, and all three isolates oxidize E2 to E1. Under anoxic conditions, CYH degrades E1 and LHJ3 degrades E2, whereas E3 and EE2 were not degraded by the three isolates; EE2 was transformed in microcosms incubated with site ground water. The degradation kinetics of E1 and E2 by CYH and E2 by LHJ3 under aerobic conditions was linearly correlated with the initial concentration, which ranged from 50 to 2,000 μg/l. The degradation of E1 by CYH under anoxic conditions followed Michaelis–Menten kinetics. 16α-Hydroxyestrone was found to be a transient transformation product of E3 under aerobic conditions.     

Recent data on intratumor estrogens in breast cancer

The contribution of local synthesis versus circulatory delivery of normal breast as well as breast cancer tissue estrogens has remained a controversial area for decades. Novel data on tissue estrogen levels confirm a positive normal breast tissue to plasma concentration gradient for estrone, and to a smaller extent estradiol. Remarkably, this gradient is similar for pre- and post-menopausal women. Together with pharmacokinetic data on estrogen disposition, these findings suggest plasma and breast tissue estrogens to rapidly equilibrate, with circulating estrogens being a major contributor to breast tissue estrone levels. A likely explanation to the concentration gradient could be the fact that non-polar estrogens easily dissolve into tissue fat compartments as compared to plasma. While intratumor estrone levels are low as compared to benign tissue concentrations, intratumor estradiol is elevated in ER+ tumors. The correlation between intratumor estradiol levels and expression levels of dehydrogenases reducing estrone into estradiol but also intratumor ER concentrations are consistent with intratumor estrogen activation but also a scavenger effect of the ER.     

Endocrine control of sexual behavior in sneaker males of the peacock blenny Salaria pavo: effects of castration, aromatase inhibition, testosterone and estradiol

The effects of castration and sex steroid manipulations on the expression of sexual behavior were investigated in a small fish, the peacock blenny, Salaria pavo. In this species, large males defend nests and attract females while small "sneaker" males reproduce by imitating the female morphology and courtship behavior in order to approach nests during spawning events and parasitically fertilize eggs. Sneakers switch into nest holders in their second breeding season, thus displaying both male and female-like sexual behavior during their lifetime. We tested the effects of castration and of an aromatase inhibitor (Fadrozole, F), testosterone (T) or 17beta-estradiol (E(2)) implants on the expression of male and female-like behavior in sneakers. Sneakers were either sham-operated, castrated or castrated and implanted with vehicle, F, T+F or E(2)+F. Seven days after the treatment, sneakers were placed in a tank with a nesting male, two ripe females and an available nest. Castrated fish had lower levels of circulating T and increased the time spent displaying female typical nuptial coloration. T implants had the opposite effect, inhibiting the expression of female-like behavior and coloration. E(2) implants had no significant effect on the display of sexual behavior but the frequency of aggressive displays decreased. The results agree with previous findings in sneakers of S. pavo that demonstrated an inhibition of female-like behavior by 11-ketotestosterone (11-KT). The reported increase in T and 11-KT production when sneakers change into nest holders may thus contribute to behaviorally defeminize sneakers. Contrarily, both T and E(2) failed to promote male-like behavior, suggesting that behavioral masculization during tactic switching depends on other neuroendocrine mechanisms or that the time length of the experiment was insufficient to induce male-like behavioral changes in sneakers.     

Do ovarian steroid hormones control the resumption of embryonic growth following the period of diapause in roe deer (Capreolus capreolus)?

Embryonic diapause in the European roe deer includes a period of five months from August to December in which embryonic development is extremely decelerated. Following exit from diapause, the embryo rapidly elongates and subsequently implants. In diapausing carnivores and marsupials, resumption of embryonic growth is regulated by ovarian steroid hormones. In the roe deer, the role of steroid hormones is not known to date. In the present study, progesterone (P4), estradiol-17β (E2) and total estrogens (E_tot) were determined in blood plasma and endometrium of roe deer shot in the course of regular huntings between September and December. Steroid hormone concentrations were correlated to the corresponding size of the embryo derived from ex vivo uterine flushing and to the date of sampling. The mean plasma concentrations of P4 (5.4 ± 0.2 ng/ml, mean ± SE, N = 87), E2 (24.3 ± 2.6 pg/ml, N = 86) and E_tot (21.7 ± 2.6 pg/ml, N = 78) remained constant over the sampling period and were not correlated to embryonic size. Likewise, endometrial concentrations of P4 (66.1 ± 6.5 ng/ml), E2 (284.0 ± 24.43 pg/ml) and, E_tot (440.9 ± 24.43 pg/ml) showed no changes over time. Therefore, it was concluded that ovarian steroid hormones do not play a determining role in resumption of embryonic growth following the period of diapause in the roe deer.     

Factors associated with estrogen receptors-alpha (ER-alpha) and -beta (ER-beta) and progesterone receptor abundance in obese and non obese pre- and post-menopausal women

There is scarce information about the factors associated with estrogen receptors (ER) at menopause. In 113 volunteers pre- and post-menopausal healthy women, grouped as with and without obesity, estrogen receptors-alpha and -beta, and progesterone receptor (PR) were measured by immunohistochemistry in skin punch biopsies obtained from the external gluteal area. In pre-menopausal women, biopsies and a blood sample were performed between days 7 and 14 of the cycle. Serum hormone levels were measured by immunoradiometric assay or radioimmunoassay. After menopause, ER and PR amounts decreased significantly. At pre-menopause, obese women had lower PR levels than non obese (P<.006). In the post-menopausal group, obese women showed higher ER-alpha (P<.03) and ER-beta (P<.02) levels than the non obese group. In the analysis of factors associated with the amount of steroid receptors for the total group, log[ER-alpha], log[ER-beta], and log[PR] were associated with age (P<.002, <.005, and <.004, respectively). The log[ER-alpha] was also associated with log[FSH] (P<.0008); meanwhile, the log[PR] showed a marginal correlation with log[FSH]. In pre-menopausal women no factor associated with any of the three receptors was found. In post-menopausal women log[ER-alpha] was associated with log[estrone] and log[DHEAS] (P<.003 and <.02, respectively). log[PR] was associated with BMI (P<.002), years since menopause (P<.05), and log[DHEAS] (P<.003). We concluded that ER and PR diminish sharply at post-menopause. At this stage the amount of receptors depends on several factors such as BMI, years since menopause, and androgen precursors.     

Ultrastructural changes of pituitary gonadotropic cells in estrogen-treated pregnant rats

Summary An ultrastructural study of gonadotropic pituitary cells was performed in estrogen-treated pregnant rats. Estradiol-treatment on Day 10 of pregnancy led to signs of ovulation or luteinization on Day 12 in 50% of the animals.Degranulation was observed in the FSH and LH cells of estrogen-responsive rats, whereas in the unresponsive group, the same cells were intensely granulated. The FSH cells of the control group showed signs of degranulation which could be correlated with follicular development. LH cells were sometimes degranulated.The role played by FSH and LH cells in the triggering of ovulation and luteinization by estrogen in the pregnant rat is discussed in the light of the ultrastructural observations.This work was financed by the DGRST, contract n^o 74.7.0030The authors wish to acknowledge the technical assistance of Mr. R. Dujol and Mr. F. Wolff     

The occurrence of estrogens in relation to reproductive processes in flowering plants

Summary Estrogens, a group of steroidal sex hormones, were determined in pollen, styles or whole pistils using a highly sensitive and specific radioimmunoassay (RIA) and found to be present at levels between 10^–11 and 10^–10 g/g FW. In the style of Liliun davidii Duch., the levels of total estrogen decreased after self-pollination of the open flower. After bud pollination, which partly overcame incompatibility, the levels of both estradiol and total estrogen increased. The levels of estradiol and total estrogen in the pistils of Brassica pekinensis Rupr. increased after compatible pollination during the first 3 days following pollination and decreased thereafter when the data were expressed on a FW basis. These results suggest that estrogens are to some extent associated with the reproductive processes.     

DNA damage induced by catecholestrogens in the presence of copper (II): generation of reactive oxygen species and enhancement by NADH

Certain estrogen metabolites are involved in carcinogenesis and the development of resistance to methotrexate (MTX). In this study, we determined whether these well-established biological effects correlate with the relative efficiency of several estrogen metabolites to induce DNA strand breaks in the presence of copper, and investigated the potential enhancing effect of reduced nicotinamide adenine dinucleotide (NADH). DNA strand breaks induced by estradiol metabolites were measured by the conversion of supercoiled phage phiX-174 RF1 DNA to open circular and linear forms. The most active catecholestrogens were the 4-hydroxy derivatives, which produced about 2.5 times more DNA double strand breaks than the 2-hydroxy derivatives, while estradiol and 16alpha-hydroxyestrone were inactive. In addition, our results show that 4-hydroxyestradiol (4-OHE2) at physiological concentrations was capable of exhibiting DNA cleaving activity. The formation of these catecholestrogen-induced DNA strand breaks was associated with the utilization of oxygen and the generation of H2O2, because catalase inhibited the DNA cleaving activity of 4-OHE2. Interestingly, we also observed that NADH enhanced the induction of DNA strands breaks by 4-OHE2/Cu(II), probably by perpetuating the redox cycle between the quinone and the semiquinone forms of the catecholestrogen. In conclusion, this study demonstrated that the relative efficiency of 2-, and 4-hydroxyestrogen in carcinogenesis and for the enhancement of MTX resistance correlates with their relative capability to induce DNA strand breaks. In order to inhibit these estrogen-mediated biological effects, it may be important to develop different strategies to block the production of reactive oxygen species by the catecholestrogen-redox cycle.     

The nucleus para-retroambiguus: a new group of estrogen receptive cells in the caudal ventrolateral medulla of the female golden hamster

Receptive female hamsters display very rigid lordotic postures. Estradiol facilitates this behavior via activation of estrogen receptors. In the hamster brainstem estrogen receptor-alpha-immunoreactive neurons (ER-alpha-IR) are present in various brainstem regions including nucleus retroambiguus (NRA) in the caudal ventrolateral medulla (CVLM) and nucleus of the solitary tract. ER-alpha-IR neurons in the CVLM project to the thoracic and upper lumbar cord. However, A1 neurons in this region do not project to the spinal cord, in contrast to overlapping C1 neurons. The question now arises: are ER-alpha-IR cells in the CVLM part of the A1/C1 group, or do they belong to the NRA or do they compose a separate cluster. A study in ovariectomized female hamsters using a combination of double immunostaining and retrograde tracing techniques and measurement of soma diameters was carried out. The results showed that A1/C1 neurons in the CVLM are almost never ER-alpha-positive; neurons inside or bordering the NRA can be divided in two different types: large multipolar and small; the large NRA-neurons, projecting caudally, are neither tyrosine hydroxylase- (TH) nor ER-alpha-IR; the small neurons, bordering the NRA and projecting caudally, are ER-alpha-IR but not TH-IR. From the available evidence and the present findings it can be concluded that the group of small ER-alpha-IR neurons in the CVLM has to be considered as a distinct entity, probably involved in the autonomic physiological changes concurring with successive phases of the estrous cycle. Because the location is closely related to the NRA itself the nucleus is called nucleus para-retroambiguus, abbreviated (NPRA).     

Brain and gonadal aromatase activity and steroid hormone levels in female and polymorphic males of the peacock blenny Salaria pavo

In the peacock blenny Salaria pavo large males with well-developed secondary sexual characters establish nests and attract females while small “sneaker” males mimic female sexual displays in order to approach the nests of larger males and parasitically fertilize eggs. These alternative reproductive tactics are sequential, as sneakers irreversibly switch into nesting males. This transition involves major morphologic and behavioral changes and is likely to be mediated by hormones. This study focuses on the role of aromatase, an enzyme that catalyses the conversion of androgens into estrogens, in the regulation of male sexual polymorphism in S. pavo. For this, sex steroid plasma levels and aromatase activity (AA) in gonads, whole brain and brain macroareas were determined in sneakers, transitional males (i.e. sneakers undergoing the transition into nesting males), nesting males and females collected in the field. AA was much higher in ovarian tissue than in testicular tissue and accordingly circulating estradiol levels were highest in females. This supports the view that elevated AA and estradiol levels are associated with the development of a functional ovary. Transitional males are in a non-reproductive phase and had underdeveloped testes when compared with sneakers and nesting males. Testicular AA was approximately 10 times higher in transitional males when compared with sneakers and nesting males, suggesting high AA has a suppressive effect on testicular development. Nesting males had significantly higher plasma levels of both testosterone (T) and 11-ketotestosterone when compared with the other male morphs and previous studies demonstrated that these androgens suppress female-like displays in sneakers. In the brain, AA was highest in macroareas presumably containing hypothalamic nuclei traditionally associated with the regulation of reproductive behaviors. Overall, females presented the highest levels of brain AA. In male morphs AA increased from sneakers, to transitional males, to nesting males in all brain macroareas. These results suggest that the transition into the nesting male tactic is accompanied both by an increase in testicular androgen production and by a higher conversion of androgens into estrogens in the brain. The increase in androgen production is likely to mediate the development of male secondary sexual characters while the increase in brain AA may be related to the behavioral changes associated with tactic transition.