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The Journal of Biological Chemistry Dec 2019Gut microbial β-glucuronidase (GUS) enzymes have been suggested to be involved in the estrobolome, the collection of microbial reactions involving estrogens....
Gut microbial β-glucuronidase (GUS) enzymes have been suggested to be involved in the estrobolome, the collection of microbial reactions involving estrogens. Furthermore, bacterial GUS enzymes within the gastrointestinal tract have been postulated to be a contributing factor in hormone-driven cancers. However, to date, there has been no experimental evidence to support these hypotheses. Here we provide the first analysis of the ability of 35 human gut microbial GUS enzymes to reactivate two distinct estrogen glucuronides, estrone-3-glucuronide and estradiol-17-glucuronide, to estrone and estradiol, respectively. We show that certain members within the Loop 1, mini-Loop 1, and FMN-binding classes of gut microbial GUS enzymes can reactivate estrogens from their inactive glucuronides. We provide molecular details of key interactions that facilitate these catalytic processes and present the structures of two novel human gut microbial GUS enzymes related to the estrobolome. Further, we demonstrate that estrogen reactivation by Loop 1 bacterial GUS enzymes can be inhibited both in purified enzymes and in fecal preparations of mixed murine fecal microbiota. Finally, however, despite these and data, we show that a Loop 1 GUS-specific inhibitor is not capable of reducing the development of tumors in the PyMT mouse model of breast cancer. These findings validate that gut microbial GUS enzymes participate in the estrobolome but also suggest that the estrobolome is a multidimensional set of processes on-going within the mammalian gastrointestinal tract that likely involves many enzymes, including several distinct types of GUS proteins.
Topics: Animals; Chromatography, High Pressure Liquid; Estrogens; Estrone; Female; Gastrointestinal Microbiome; Glucuronidase; Male; Mice; Mice, Inbred BALB C; Mutagenesis, Site-Directed
PubMed: 31636122
DOI: 10.1074/jbc.RA119.010950 -
Blood Mar 2022
Topics: Estrone; Fibrin; Fibrinogen
PubMed: 35238891
DOI: 10.1182/blood.2021015215 -
Physiological Reports Oct 2023Our current understanding of the relationship between estrogen and human endothelial colony-forming cell (hECFC) function is based almost exclusively on studies...
Our current understanding of the relationship between estrogen and human endothelial colony-forming cell (hECFC) function is based almost exclusively on studies investigating estradiol action at nuclear estrogen receptors. In the current study the hypothesis was tested that the less potent estrogen receptor agonist, estrone, affects hECFC proliferation, migration, secretion, and tube formation in a way that is unique from that of estradiol. The relationship between the estrogens, estradiol and estrone, is clinically important, particularly in postmenopausal women where estradiol levels wane and estrone becomes the predominant estrogen. Cultured hECFCs from peripheral blood mononuclear cell fractions were treated with concentrations of estradiol and estrone ranging from 1 nM to 1 μM separately and in combination. Following treatment, proliferation, migration, ability to attract other hECFCs (autocrine secretion), and ability to enhance endothelial cell tube formation (tubulogenesis) were tested. Functional assays revealed unique, concentration-dependent physiological effects of estrone and estradiol. Estradiol exposure resulted in increased hECFC proliferation, migration, secretion of chemoattractant, and enhancement of tube formation as expected. As with estradiol, hECFC secretion of chemoattractant increased significantly with each increase in estrone exposure. Estrone treatment produced a biphasic, concentration-dependent relationship with proliferation and tube formation and relatively no effect on hECFC migration at any concentration. The quantitative relationship between the effects of estrone and estradiol and each hECFC function was analyzed. The extent to which estrone was similar in effect to that of estradiol was dependent on both the concentrations of estradiol and estrone and the hECFC function measured. Interestingly, when the two estrogens were present, differing ratios resulted in unique functional responses. hECFCs that were treated with combinations of estrone and estradiol with high estrone to estradiol ratios showed decreased proliferative capacity. Conversely, hECFCs that were treated with combinations that were relatively high in estradiol, showed increased proliferative capacity. Cells that were treated with estrone and estradiol in equal concentrations showed an attenuated proliferative response that was decreased compared to the proliferation that either estrone or estradiol produced when they were present alone. This co-inhibitory relationship, which has not been previously reported, challenges the prevailing understanding of estrone as solely a weak agonist at estrogen receptors. This study provides evidence that estrone signaling is distinct from that of estradiol and that further investigation of estrone's mechanism of action and the biological effect may provide important insight into understanding the dysfunction and decreased number of hECFCs, and the resulting cardiovascular disease risk observed clinically in menopausal women and women undergoing hormone replacement therapy.
Topics: Female; Humans; Estrone; Estradiol; Receptors, Estrogen; Leukocytes, Mononuclear; Estrogens; Endothelial Cells; Chemotactic Factors
PubMed: 37792856
DOI: 10.14814/phy2.15818 -
The Science of the Total Environment Feb 2021Estrone and BPA are two endocrine disrupting chemicals (EDCs) that are predicted to be less potent than estrogens such as 17β-estradiol and 17α-ethinylestradiol. Human...
Estrone and BPA are two endocrine disrupting chemicals (EDCs) that are predicted to be less potent than estrogens such as 17β-estradiol and 17α-ethinylestradiol. Human exposure concentrations to estrone and BPA can be as low as nanomolar levels. However, very few toxicological studies have focused on the nanomolar-dose effects. Low level of EDCs can potentially cause non-monotonic responses. In addition, exposures at different developmental stages can lead to different health outcomes. To identify the nanomolar-dose effects of estrone and BPA, we used zebrafish modeling to study the phenotypic and transcriptomic responses after extended duration exposure from 0 to 5 days post-fertilization (dpf) and short-term exposure at days 4-5 post fertilization. We found that non-monotonic transcriptomic responses occurred after extended duration exposures at 1 nM of estrone or BPA. At this level, estrone also caused hypoactivity locomotive behavior in zebrafish. After both extended duration and short-term exposures, BPA led to more apparent phenotypic responses, i.e. skeletal abnormalities and locomotion changes, and more significant transcriptomic responses than estrone exposure. After short-term exposure, BPA at concentrations equal or above 100 nM affected locomotive behavior and changed the expression of both estrogenic and non-estrogenic genes that are linked to neurological diseases. These data provide gaps of mechanisms between neurological genes expression and associated phenotypic response due to estrone or BPA exposures. This study also provides insights for assessing the acceptable concentration of BPA and estrone in aquatic environments.
Topics: Animals; Benzhydryl Compounds; Endocrine Disruptors; Estrone; Humans; Phenols; Transcriptome; Zebrafish
PubMed: 33243503
DOI: 10.1016/j.scitotenv.2020.143736 -
Frontiers in Endocrinology 2022To discover the profiles of different steroid hormones at the maternal-fetal interface and reveal the change characteristics in pregnant women at advanced maternal age...
OBJECTIVES
To discover the profiles of different steroid hormones at the maternal-fetal interface and reveal the change characteristics in pregnant women at advanced maternal age (AMA).
METHODS
Forty pregnant women were recruited in the study, including 20 AMA women (age ≥ 35) and 20 normal controls (age < 35 and without pregnancy complications). Among AMA women, 6 (AMA2) had pregnancy complications, and 14 (AMA1) had no complications. Their maternal blood (MB), placental tissue (P), and fetal cord blood (CB) were collected, and 18 different steroid hormone metabolites were analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS).
RESULTS
The estradiol (E2) levels in MB were higher than those in P and CB. In contrast, the estrone (E1) and estriol (E3) levels were higher in P and CB. Compared with the progesterone levels (P4) in MB, those in P and CB were higher; however, cortisol (F) levels were deficient. In contrast, F in MB was maintained at an elevated level. Further, cortisone (E) levels in CB were higher than those in MB and P. Except for the decline of testosterone (T), androstenedione (A2) and Dihydrotestosterone (DHT), there were no significant differences in the other 15 steroid hormones in MB between the AMA1 and the control group (p>0.05). Compared with the AMA1 group, androgen levels were significantly higher in AMA2, especially in T (1.55 0.68 ng/ml, p=0.023), A2 (2.27 0.92 ng/ml, p=0.011) and Dehydroepiandrosterone (DHEA) (2.39 1.50 ng/ml, p=0.028). However, there were no significant changes in P and CB between two groups.
CONCLUSION
There are distribution rules and cascade changes of steroid profiles in maternal-fetal compartments. Significantly high androgen levels in AMA women have a positive relationship with adverse pregnancy complications.
Topics: Androgens; Chromatography, Liquid; Estrone; Female; Humans; Placenta; Pregnancy; Pregnancy Complications; Pregnant Women; Tandem Mass Spectrometry
PubMed: 35282454
DOI: 10.3389/fendo.2022.796909 -
Molecules (Basel, Switzerland) May 2021The interest in the introduction of the oxime group in molecules aiming to improve their biological effects is increasing. This work aimed to develop new steroidal...
The interest in the introduction of the oxime group in molecules aiming to improve their biological effects is increasing. This work aimed to develop new steroidal oximes of the estrane series with potential antitumor interest. For this, several oximes were synthesized by reaction of hydroxylamine with the 17-ketone of estrone derivatives. Then, their cytotoxicity was evaluated in six cell lines. An estrogenicity assay, a cell cycle distribution analysis and a fluorescence microscopy study with Hoechst 3358 staining were performed with the most promising compound. In addition, molecular docking studies against estrogen receptor α, steroid sulfatase, 17β-hydroxysteroid dehydrogenase type 1 and β-tubulin were also accomplished. The 2-nitroestrone oxime showed higher cytotoxicity than the parent compound on MCF-7 cancer cells. Furthermore, the oximes bearing halogen groups in A-ring evidenced selectivity for HepaRG cells. Remarkably, the Δ-estrone oxime was the most cytotoxic and arrested LNCaP cells in the G/M phase. Fluorescence microscopy studies showed the presence of condensed DNA typical of prophase and condensed and fragmented nuclei characteristic of apoptosis. However, this oxime promoted the proliferation of T47-D cells. Interestingly, molecular docking studies estimated a strong interaction between Δ-estrone oxime and estrogen receptor α and β-tubulin, which may account for the described effects.
Topics: Cell Cycle; Cell Death; Cell Line, Tumor; Cell Proliferation; Cell Survival; DNA; Estrogen Receptor alpha; Estrogens; Estrone; Fluorouracil; Humans; Inhibitory Concentration 50; Molecular Docking Simulation; Oximes
PubMed: 34064380
DOI: 10.3390/molecules26092687 -
Seminars in Thrombosis and Hemostasis Mar 2022Fibrinogen, one of the most abundant plasma proteins playing a key role in hemostasis, is an important modulator of wound healing and host defense against microbes. In... (Review)
Review
Fibrinogen, one of the most abundant plasma proteins playing a key role in hemostasis, is an important modulator of wound healing and host defense against microbes. In the current review, we address the role of fibrin(ogen) throughout the process of wound healing and subsequent tissue repair. Initially fibrin(ogen) acts as a provisional matrix supporting incoming leukocytes and acting as reservoir for growth factors. It later goes on to support re-epithelialization, angiogenesis, and fibroplasia. Importantly, removal of fibrin(ogen) from the wound is essential for wound healing to progress. We also discuss how fibrin(ogen) functions through several mechanisms to protect the host against bacterial infection by providing a physical barrier, entrapment of bacteria in fibrin(ogen) networks, and by directing immune cell function. The central role of fibrin(ogen) in defense against bacterial infection has made it a target of bacterial proteins, evolved to interact with fibrin(ogen) to manipulate clot formation and degradation for the purpose of promoting microbial virulence and survival. Further understanding of the dual roles of fibrin(ogen) in wound healing and infection could provide novel means of therapy to improve recovery from surgical or chronic wounds and help to prevent infection from highly virulent bacterial strains, including those resistant to antibiotics.
Topics: Estrone; Fibrin; Fibrinogen; Humans; Infection Control; Wound Healing
PubMed: 34428799
DOI: 10.1055/s-0041-1732467 -
Journal of Dairy Science Jun 2010Some individuals fear that estrogens in dairy products may stimulate growth of estrogen-sensitive cancers in humans. The presence of estrone (E(1)) and 17beta-estradiol...
Some individuals fear that estrogens in dairy products may stimulate growth of estrogen-sensitive cancers in humans. The presence of estrone (E(1)) and 17beta-estradiol (E(2)) in raw whole cow's milk has been demonstrated. The objectives of this study were to determine if pasteurization-homogenization affects E(2) concentration in milk and to quantify E(1) and E(2) concentrations in commercially available dairy products. The effects of pasteurization-homogenization were tested by collecting fresh raw milk, followed by pasteurization and homogenization at 1 of 2 homogenization pressures. All treated milks were tested for milk fat globule size, percentages of milk fat and solids, and E(2) concentrations. Estrone and E(2) were quantified from organic or conventional skim, 1%, 2%, and whole milks, as well as half-and-half, cream, and butter samples. Estrone and E(2) were quantified by RIA after organic solvent extractions and chromatography. Pasteurization-homogenization reduced fat globule size, but did not significantly affect E(2), milk fat, or milk solids concentrations. Estrone concentrations averaged 2.9, 4.2, 5.7, 7.9, 20.4, 54.1 pg/mL, and 118.9 pg/g in skim, 1%, 2%, and whole milks, half-and-half, cream, and butter samples, respectively. 17Beta-estradiol concentrations averaged 0.4, 0.6, 0.9, 1.1, 1.9, 6.0 pg/mL, and 15.8 pg/g in skim, 1%, 2%, whole milks, half-and-half, cream, and butter samples, respectively. The amount of fat in milk significantly affected E(1) and E(2) concentrations in milk. Organic and conventional dairy products did not have substantially different concentrations of E(1) and E(2). Compared with information cited in the literature, concentrations of E(1) and E(2) in bovine milk are small relative to endogenous production rates of E(1) and E(2) in humans.
Topics: Animals; Cattle; Dairy Products; Estradiol; Estrone; Female; Food Preservation; Milk; Radioimmunoassay
PubMed: 20494161
DOI: 10.3168/jds.2009-2947 -
Theriogenology Sep 2022This study describes 17-β-estradiol (E2), estrone (E1) and estrone-sulfate (E1S) concentrations between 4 and 11 months in healthy equine pregnancies of two different...
This study describes 17-β-estradiol (E2), estrone (E1) and estrone-sulfate (E1S) concentrations between 4 and 11 months in healthy equine pregnancies of two different breeds using Liquid Chromatography coupled to Mass-Spectrometry (LC-MS). In 2 stud-farms including 15 Spanish PureBred (SPB) and 11 Showjumping (SJ) types mares, combined thickness of the uterus and the placenta (CTUP) was measured and blood was sampled monthly between 4 and 11 months of gestation. Concentrations of E2, E1 and E1S were assayed with LC-MS in mares with normal CTUP. Effects of breed, day of pregnancy and mare's parity and age on estrogens concentrations were investigated. Peak of E2 was observed at 5 months (median: 46.4 pg/mL; maximum: 201.5 pg/mL). A strong correlation was observed between E1 and E1S (p < 0.0001, r = 0.85). Peak of E1 (median: 571.0 pg/mL; maximum: 1641.9 pg/mL) and E1S (median: 573.6 ng/mL; maximum: 997.6 ng/mL) concentrations was observed at the 5th month and then E1S decreased quicker than E1 until the end of pregnancy. Higher E2 and E1 concentrations were observed in SJ than in SPB mares between the 6th and the 8th months. No difference between breeds was observed for E1S monthly evolution. Estrogen peak values were all observed at 5 months. Unlike recent LC-MS studies, E1S values observed here were in the same range than those previously established using immuno-assays. After the 6th month, E1S decreased quicker than E1. Effect of breed only observed on non-sulfonated estrogens should be further confirmed. These findings confirm that sulfonation activity of the allantochorion may be limited after the 6th month.
Topics: Animals; Chromatography, Liquid; Estradiol; Estrogens; Estrone; Female; Horses; Mass Spectrometry; Pregnancy; Sulfates
PubMed: 35738034
DOI: 10.1016/j.theriogenology.2022.06.004 -
Research in Veterinary Science May 2021Steroid concentrations in serum are fluctuating during pregnancy of many mammal species. The current knowledge about endocrinology of gestation is mainly based on...
Development and validation of a liquid chromatography coupled to mass spectrometer (LC-MS) method for the simultaneous quantification of estrone-3-sulfate, progesterone, estrone and estradiol in serum of mares and American bisons.
Steroid concentrations in serum are fluctuating during pregnancy of many mammal species. The current knowledge about endocrinology of gestation is mainly based on immunoassays. However, the lack of specificity of these assays hampers the reliability of the results. In the present work, we developed and validated a methodology associating liquid chromatography (LC) and mass spectrometry (MS) to simultaneously quantify, with high specificity and accuracy, estrone-3-sulfate (E3S), progesterone (PRO), estrone (E1) and estradiol (E2) in serum of two different mammal species. The sample preparation procedure is based on a simple protein precipitation and a derivatization with dansyl chloride. After the chromatographical separation, compounds were analyzed with a triple-quadrupole mass spectrometer operating in multiple reaction monitoring. Mare and American bison serum samples were analyzed with the validated method and results were compared with concentrations measured with commercial radioimmunoassay (RIA), enzyme linked immunosorbent assay (ELISA) and chemiluminescent microparticle immunoassay (CMIA). Following these criterions: relative standard deviation <15% and relative bias <15%, lower limits of quantification of 0.5 ng/mL (E3S), 0.1 ng/mL (PRO) and 2 pg/mL (E1 and E2) were achieved. Most of the comparison between immunoassays and LC-MS showed poor correlation and proportional differences. Our LC-MS method is able to simultaneously quantify several steroid hormones with high specificity, accuracy and sensitivity in serum of two different mammal species. Our method constitutes a useful and performant tool for veterinary clinicians and LC-MS should thus be used to update and refine the current knowledge about the endocrinology of pregnancy in mammals.
Topics: Animals; Bison; Chromatography, Liquid; Estradiol; Estrone; Female; Horses; Pregnancy; Progesterone; Reproducibility of Results; Tandem Mass Spectrometry; United States
PubMed: 33770524
DOI: 10.1016/j.rvsc.2021.03.014