Diethylstilbestrol Research Articles

Transcriptional profiling of the developing rat ovary following intrauterine exposure to the endocrine disruptors diethylstilbestrol and ketoconazole

Exposure to endocrine-disrupting chemicals (EDCs) during development may cause reproductive disorders in women. Although female reproductive endpoints are assessed in rodent toxicity studies, a concern is that typical endpoints are not sensitive enough to detect chemicals of concern to human health. If so, measured endpoints must be improved or new biomarkers of effects included. Herein, we have characterized the dynamic transcriptional landscape of developing rat ovaries exposed to two well-known EDCs, diethylstilbestrol (DES) and ketoconazole (KTZ), by 3’ RNA sequencing. Rats were orally exposed from day 7 of gestation until birth, and from postnatal day 1 until days 6, 14 or 22. Three exposure doses for each chemical were used: 3, 6 and 12 µg/kg bw/day of DES; 3, 6, 12 mg/kg bw/day of KTZ. The transcriptome changed dynamically during perinatal development in control ovaries, with 1137 differentially expressed genes (DEGs) partitioned into 3 broad expression patterns. A cross-species deconvolution strategy based on a mouse ovary developmental cell atlas was used to map any changes to ovarian cellularity across the perinatal period to allow for characterization of actual changes to gene transcript levels. A total of 184 DEGs were observed across dose groups and developmental stages in DES-exposed ovaries, and 111 DEGs in KTZ-exposed ovaries across dose groups and developmental stages. Based on our analyses, we have identified new candidate biomarkers for female reproductive toxicity induced by EDC, including Kcne2, Calb2 and Insl3.

Identification of biomarkers and outcomes of endocrine disruption in human ovarian cortex using In Vitro Models

Endocrine disrupting chemicals (EDCs) are raising concerns about adverse effects on fertility in women. However, there is a lack of information regarding mechanisms and effects in humans. Our study aims to identify mechanisms of endocrine disruption using two EDCs, diethylstilbestrol (DES) and ketoconazole (KTZ)11Ketoconazole: KTZ.. Human ovarian cortical tissue obtained from Caesarean section patients was exposed to 10−9 M - 10−5 M KTZ and 10−10 M - 10−6 M DES in vitro for 6 days. Follicle survival and growth were studied via histology analysis and liquid-chromatography-mass spectrometry-based steroid quantification. RNA-sequencing was performed on COV434, KGN, and primary ovarian cells that were exposed for 24 h. Significantly lower unilaminar follicle densities were observed in DES 10−10 M group, whereas low KTZ exposure reduced secondary follicle density. KTZ 10−5 M reduced levels of pregnenolone and progesterone. RNA-sequencing revealed that 445 and 233 differentially expressed genes (false discovery rate < 0.1) altogether in DES and KTZ exposed groups. Gene set variation analysis showed that both chemicals modulated pathways that are important for folliculogenesis and steroidogenesis. We selected stearoyl-CoA desaturase (SCD) and 7-dehydrocholesterol reductase (DHCR7) for further validation. Up-regulation of both genes in response to KTZ was confirmed by qPCR and in situ RNA hybridization. Further validation with immunofluorescence focused on the expression of SCD in growing follicles in exposed ovarian tissue. In conclusion, SCD may serve as a potential novel human-relevant biomarker of EDC exposure and effects on ovaries.

Human papilloma virus associated primary vaginal adenocarcinoma

Primary vaginal adenocarcinoma associated with human papillomavirus (HPV) infection is extremely rare. We report a case of primary adenocarcinoma of the vagina associated with human papilloma virus successfully treated with anterior pelvic exenteration and adjuvant concurrent chemoradiation therapy. A 51-year-old postmenopausal woman (gravida 1, para 1) presented with intermittent vaginal bleeding and pelvic pain. She was found to have a 5 × 5 cm necrotic tumor took up the vaginal. She had no previous history of antenatal exposure to diethylstilbestrol (DES). Pelvic magnetic resonance imaging (MRI) demonstrated a 4.8 × 6.0 cm mass in the vaginal canal, an 1.1 × 2.6 cm mass at the urinary bladder dome and a 1.1 cm irregular lymph node at the right external iliac chain with increased fluorodeoxyglucose (FDG) uptake from Fused whole-body positron emission tomography-computed tomography (PET-CT). Based on clinical investigations, the patient was diagnosed with a primary adenocarcinoma of vagina, staged International Fedestration of Gynecology and Obstetrics (FIGO) IVa. Anterior pelvic exenteration, simple vulvectomy, total vaginectomy, both pelvic lymph node dissection, and para-aortic lymph node dissection with ileal conduit urinary diversion (Bricker’s operation) was done. Histologically primary vaginal HPV type 16-associated adenocarcinoma was confirmed. Both obturator lymph node was positive for metastasis. Postoperatively, the patient received weekly cisplatin regimen administered with a dose of 40 mg/m2 on day 1 of external radiation therapy (RT), 1 to 4 hours before RT initiation. External beam pelvic RT dose prescription to the whole pelvis was 59.4 Gy in 33 fractions at the isocenter. But, after total dose of 43.2 Gy, patient complained severe bowel habit change and discontinued further treatment. The patient remains free from recurrence 8 months after initial surgery. In the lack of information and comparative analysis of management options for the more unusual and rare varieties of primary vaginal neoplasms in the literature, this suggests the possibility that surgical treatment may be preferentially selected on a case-by-case basis.

Tralucent and Pellucid-Clear Cell Adenocarcinoma Uterine Cervix

Clear cell adenocarcinoma of uterine cervix is predominantly comprised of clear cells or hobnail cells articulating distinct architectural patterns as solid, tubulocystic or papillary. Tumour cell cytoplasm is clear, eosinophilic with intracytoplasmic hyaline globules and enhanced nuclear grade. A historic association with intrauterine diethylstilbestrol (DES) exposure is enunciated. Sporadic overexpression of p53 tumour suppressor gene may be observed. Neoplasm manifests with vaginal bleeding, postcoital bleeding or abnormal vaginal discharge. Cytological smears exhibit sheets, clusters or papillae of tumour cells permeated with delicate, vacuolated, glycogen rich or finely granular cytoplasm, enlarged, pale, spherical nuclei with prominent nucleoli or naked nuclei with tigroid background composed of granular, reticulated substance. Clear cell adenocarcinoma appears immune reactive to pan-cytokeratin as AE1 / AE3, CAM5.2, CK7, CK8, CK18, CK19, HNF1β, Napsin A, Ki67 or epithelial membrane antigen (EMA). Tumefaction requires segregation from neoplasms such as microglandular hyperplasia, Arias-Stella reaction, primary alveolar soft part sarcoma, mesonephric adenocarcinoma, metastatic clear cell renal cell carcinoma, endometrioid carcinoma, serous carcinoma, squamous cell carcinoma, urothelial carcinoma, adenosis, endometrioid adenocarcinoma with clear cell or secretory change, gastric subtype of endocervical adenocarcinoma or yolk sac tumours of uterine cervix. Neoplasm may be preliminarily subjected to radical hysterectomy with pelvic lymphadenectomy or fertility sparing radical trachelectomy.

Anti-ovarian cancer potential, in silico studies, and anti-Alzheimer's disease effects of some natural compounds as cholinesterase inhibitors.

Ovarian cancer ranks seventh in the most common malignant tumors in females and seriously threatens women's reproductive health. Natural sources may lead to basic research on potential bioactive components as lead compounds in drug discovery and, ultimately, therapeutic treatments for ovarian cancer and other diseases. Alzheimer's disease (AD) and ovarian cancer are complex diseases of aging that impose an enormous public health burden worldwide. Additionally, people with AD have low levels of acetylcholine in their brains. Enzymes called cholinesterases break down acetylcholine in the brain. If their action is inhibited, more acetylcholine is available for communication among brain cells. In this study, pregnanolone, diethylstilbestrol (DES), flavokawain C, and methyl 3,4,5-trihydroxybenzoate molecules obtained excellent-to-good inhibitory against acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) enzymes with IC50 values ranging between 77.18±8.62 to 461.35±28.54μM for AChE and 23.86±4.07 to 306.62±32.46μM for BuChE. The calculations revealed the probable interactions and their characteristics at an atomic level. Indeed, the docking scores of DES, flavokawain C, pregnanolone, and methyl 3,4,5-trihydroxybenzoate for AChE are -6.685, -6.247, -6.672, and -5.183(kcal/mol), respectively. This value for the compounds against BuChE is -6.042, -8.851, -5.655, and -5.898(kcal/mol), respectively. Additionally, these compounds significantly decreased ovarian cancer cell viability. Additionally, 100μM dose of all molecules caused good reductions in ovarian cancer cell viability.

Endocrine disrupting effect and reproductive toxicity of the separate exposure and co-exposure of nano-polystyrene and diethylstilbestrol to zebrafish

The co-occurrence of nanoplastics and other pollutants in the environment has gotten a lot of attention, but information on the biological toxicity of their co-exposure was limited. This study aims to reveal the endocrine disrupting effect and reproductive toxicity of nano-polystyrene (NPS) and diethylstilbestrol (DES) to zebrafish under separate and combined exposure. Results indicated that NPS and DES exposure in isolation reduced the hepatosomatic index and gonadosomatic index, and altered the cell maturity in gonads in both cases. Even worse, the co-exposure of NPS and DES exacerbated the damage to the liver and gonads of fish. The two pollutants individually inhibited the secretion of sex hormones and vitellogenin. The inhibition effect of DES was especially dose-dependent, while NPS had weaker effect than DES. Their combined action on the secretion of sex hormones and vitellogenin exhibited additive effect. However, NPS did not affect the content of thyroid hormones in fish, and also had no significant effect on the reduction of thyroid hormone caused by DES exposure. Furthermore, their co-exposure decreased the cumulative eggs from 1031 to 306, and the spawning number from 12 to 8. The fertilization rate and hatchability rete of eggs were reduced by 30.9% and 40.4%, respectively. The abnormality rate of embryos was 65.0%, significantly higher than in separate DES and NPS groups (55.7% and 30.8% respectively). The abnormal development of offspring was mainly pericardial cyst, spinal curvature, and growth retardation.

Primary Clear Cell Adenocarcinoma of the Uterine Cervix in a 14-Year-Old Virgin Girl: Case Report

Cervical cancer is rare in adolescent and pediatric populations, with adenocarcinoma being the most commonly reported. Clear cell adenocarcinoma of the uterine cervix accounts for only 4% of all adenocarcinoma cases, and about two-thirds are associated with intrauterine diethylstilbestrol (DES) exposure. We report the case of a 14-year-old virgin girl who presented with a 1-month-long history of abnormal vaginal bleeding and lower abdominal pain. Transabdominal pelvic ultrasound examination revealed the presence of an irregular, homogeneous cervical mass that was 7 cm in size. Therefore, a magnetic resonance imaging (MRI) scan was performed to establish the origin of the tumor and its relationship to adjacent pelvic organs. Furthermore, a vaginoscopy was performed to identify the tumor, and a cervical biopsy was performed. Immunohistochemical and anatomopathological studies resulted in the diagnosis of non-HPV(Human Papilloma Virus)-related clear cell adenocarcinoma of the cervix. Following the oncological examination, she was admitted for radiotherapy. The patient had no maternal history of DES exposure in utero. Even though the number of cases in the literature is low, most of the virgin girls diagnosed with clear cell adenocarcinoma of the cervix have a fatal prognosis because of the delay in making a correct diagnosis.

Maintaining bone health by estrogen therapy in patients with advanced prostate cancer: a narrative review.

The purpose of androgen deprivation therapy (ADT) in prostate cancer (PCa), using luteinizing hormone-releasing hormone agonists (LHRHa) or gonadotrophin-releasing hormone antagonists, is to suppress the levels of testosterone. Since testosterone is the precursor of estradiol (E2), one of the major undesired effects of ADT is the concomitant loss of E2, causing among others an increased bone turnover and bone loss and an increased risk of osteoporosis and fractures. Therefore, the guidelines for ADT indicate to combine ADT routinely with bone-sparing agents such as bisphosphonates, denosumab or selective estrogen receptor modulators. However, these compounds may have side effects and some require inconvenient parenteral administration. Co-treatment with estrogens is an alternative approach to prevent bone loss and at the same time, to avoid other side effects caused by the loss of estrogens, which is the topic explored in the present narrative review. Estrogens investigated in PCa patients include parenteral or transdermal E2, diethylstilbestrol (DES), and ethinylestradiol (EE) as monotherapy, or high-dose estetrol (HDE4) combined with ADT. Cardiovascular adverse events have been reported with parenteral E2, DES and EE. Encouraging effects on bone parameters have been obtained with transdermal E2 (tE2) and HDE4, in the tE2 development program (PATCH study), and in the LHRHa/HDE4 co-treatment study (PCombi), respectively. Confirmation of the beneficial effects of estrogen therapy with tE2 or HDE4 on bone health in patients with advanced PCa is needed, with special emphasis on bone mass and fracture rate.

Potential Adsorption Affinity of Estrogens on LDPE and PET Microplastics Exposed to Wastewater Treatment Plant Effluents.

Microplastics (MPs) are among the most common pollutants in the environment. Because of their small size, availability, and similarity to natural foods, they are commonly ingested by marine organisms. They can cause health problems in living organisms due to their bioaccumulation potential. It is, therefore, unknown whether endocrine-disrupting chemicals (EDCs), in particular estrogens, are capable of adhering to the diverse types of MPs found in water. Two MP polymers (low-density polyethylene (LDPE) and polyethene terephthalate (PET)) that could pose a threat to fish were tested for estrogen adsorption. The adsorption capacity of MP pellets was studied for 30 days in the effluent and influent of a wastewater treatment-plant. A laboratory simulation was conducted to validate the field and laboratory findings. We found that the concentrations of five types of estrogen ((diethylstilbestrol (DES), estrone (E1), 17β-estradiol (E2), estriol (E3), 17α-ethinyl estradiol (EE2)) were higher in the influent than the effluent streams. LDPE and PET MPs exposed to influent water in the laboratory had higher estrogen levels than wastewater treatment plants (WWTP) pellets. The PET pellets showed the highest adsorption affinity to EE2, while the LDPE pellets showed the highest affinity to E2. As a result, this study provided baseline data to investigate the estrogen adsorption capacity in MPs.

Oxidation of contaminants of emerging concern by combination of peracetic acid with iron ions and various types of light radiation – Optimization, kinetics, removal efficiency and mechanism investigation

The aim of this work was to check the effectiveness of PAA activated by fluorescent light low intensive radiation, UV radiation (254 or 366 nm), simulated solar light and also in PAA-iron (Fe2+ or Fe3+) ions in the degradation of selected contaminants of emerging concern (CECs). The kinetics of the degradation process of benzophenone (BPH), 4-n-octylphenol (4OP), 4-n-nonylphenol (4NP), 3-(4-methylbenzylidene) camphor (MBC), endosulfan I (ESI), endosulfan II (ESII), oxybenzone (OXB), bisphenol A (BPA), diethylstilbestrol (DSB) and estrone (E1) in such systems have been studied. Using chemometric technique design of experiments (DoE) the following composition of PAA-H2O2 equilibrated mixture was selected as optimal: CPAA = 10.5 mgL-1, CH2O2 = 20 mgL-1 and pH = 6. The most effective appeared to be PAA-Fe(III) oxidizing system. High degradation efficiency, above 90 %, was observed for all tested compounds in the presence of Fe (III) ions as an activator. Using the EPR technique, the radical nature of the processes under study was proved. The influence of a matrix (raw and purified wastewater) on the degradation rate has been checked. Insight into the process done with Density Functional Theory (DFT) showed that ∙OH addition and H abstraction are the main mechanisms initiating analytes degradation.

Occurrence and Degradation of Free and Conjugated Estrogens in a River Receiving Feedlot Animal Discharge

This study analyzed concentrations of 17β-estradiol (E2), estrone (E1), estriol (E3), 17α-ethynylestradiol (EE2), diethylstilbestrol (DES), 17β-estradiol-3-sulfate (E2-3S), estrone-3-sulfate (E1-3S), 17β-estradiol-3-glucuronide (E2-3G), and estrone-3-glucuronide (E1-3G) in river water, received from intensive feedlot operations wastewater in WuLo Creek, Taiwan. Moreover, the estrogen degradation in situ was analyzed. The average concentrations were 54.15 ± 31.42, 9.71 ± 6.42 and 3.55 ± 2.41 ng/L for E1, E2 and E3, respectively. The concentrations and order were similar to the polluted river and higher than most rivers’ concentrations. The conjugated estrogen concentrations ranged from ND to 13.2 ng/L (E1-3S), ND to 10.4 ng/L (E2-3S), ND to 10.0 ng/L (E1-3G), and ND to 3.6 ng/L (E2-3G), and the detection rates were 76%, 71%, 56%, and 15%, respectively. In the present study, the high detection rates of conjugate estrogen were more elevated than the water receiving STP effluent, suggesting that the source was the river water close to the animal wastewater discharge. In the degradation test, the DES concentrations slightly declined after 24 h, but E1-3G did not significantly change over time (p &gt; 0.05). The degradation of free estrogen occurred during the first 12 h period, and residual concentration was not further decreased after 24 h. In the environment, E1 had higher concentrations than E2 and E3, suggesting that E1 was more resistant to degradation than E2 and E3 at low concentrations. However, the degradation test in the present study suggested that E1 rapidly degraded at high ambient concentrations due to the high degradation constant.

Diethylstilbestrol and autism.

It is acknowledged that diethylstilbestrol (DES), a synthetic diphenol with powerful estrogenic properties, causes structural anomalies of the reproductive tract and increases the risk of cancer and genital malformations in children and grandchildren of mothers treated during pregnancy. Conversely, data on DES effects on neurodevelopment and psychiatric disorders in in-utero exposed children and their descendants are rare, especially concerning Autism Spectrum Disorders (ASD). Recent studies presented in this review strengthen the hypothesis that in-utero exposure to DES and also other synthetic estrogens and progestogens, which all are endocrine disruptors, contributes to the pathogenesis of psychiatric disorders, especially ASD. A large epidemiological study in the USA in 2010 reported severe depression in in-utero exposed children (n=1,612), and a French cohort study (n=1,002 in-utero DES exposed children) in 2016 found mainly bipolar disorders, schizophrenia, major depression, suicide attempts, and suicide. Few publications described ASD in in-utero exposed children, mainly a Danish cohort study and a large Chinese epidemiological study. Molecular studies on endocrine disruptors demonstrated the transgenerational induction of diseases and DES epigenetic impact (DNA methylation changes) at two genes implicated in neurodevelopment (ZFP57 and ADAM TS9). We recently described in an informative family, somatic and psychiatric disorders in four generations, particularly ASD in boys of the third and fourth generation. These data show that the principle of precaution must be retained for the protection of future generations: women (pregnant or not) should be extremely vigilant about synthetic hormones.

Multiple tissue-specific epigenetic alterations regulate persistent gene expression changes following developmental DES exposure in mouse reproductive tissues

ABSTRACT Clinically, developmental exposure to the endocrine disrupting chemical, diethylstilboestrol (DES), results in long-term male and female infertility. Experimentally, developmental exposure to DES results in abnormal reproductive tract phenotypes in male and female mice. Previously, we reported that neonatal DES exposure causes ERα-mediated aberrations in the transcriptome and in DNA methylation in seminal vesicles (SVs) of adult mice. However, only a subset of DES-altered genes could be explained by changes in DNA methylation. We hypothesized that alterations in histone modification may also contribute to the altered transcriptome during SV development. To test this idea, we performed a series of genome-wide analyses of mouse SVs at pubertal and adult developmental stages in control and DES-exposed wild-type and ERα knockout mice. Neonatal DES exposure altered ERα-mediated mRNA and lncRNA expression in adult SV, including genes encoding chromatin-modifying proteins that can impact histone H3K27ac modification. H3K27ac patterns, particularly at enhancers, and DNA methylation were reprogrammed over time during normal SV development and after DES exposure. Some of these reprogramming changes were ERα-dependent, but others were ERα-independent. A substantial number of DES-altered genes had differential H3K27ac peaks at nearby enhancers. Comparison of gene expression changes, H3K27ac marks and DNA methylation marks between adult SV and adult uterine tissue from ovariectomized mice neonatally exposed to DES revealed that most of the epigenetic changes and altered genes were distinct in the two tissues. These findings indicate that the effects of developmental DES exposure cause reprogramming of reproductive tract tissue differentiation through multiple epigenetic mechanisms.

Determination of Diethylstilbestrol in Environmental Water Based on Co(II)-Sensitized Electrochemical Sensing

In this research, a simple electrochemical sensor (c-MWCNTs/Nafion/GCE) was prepared by carboxylated multi-walled carbon nanotube, perfluorosulfonic acid polymer and glassy carbon electrode using drop coating method, and based on the obvious sensitization effect on the redox peak current of diethylstilbestrol (DES) in the presence of Co2+, a new electrochemical sensing method for the determination of DES in water was established. In a buffered solution of 0.2 mol l−1 NaH2PO4-Na2HPO4 at pH = 7.0, differential pulse voltammetry (DPV) was used when the Co2+ concentration was 20 μmol l−1. Under the optimized conditions of −0.2 ∼ −0.4 V (vs SCE), −0.2 V enrichment potential, and 120 s enrichment time, the concentration of DES showed a good linear relationship with the peak current over the range of 0.3 to 90 μmol l−1, and the detection limit(S/N = 3) was 0.01 μmol l−1. The actual water sample was detected, and the recovery was between 89.17% and 101.23%. This method had the characteristics of simple operation, low analysis cost and fast analysis speed, and had the potential application value of detecting DES in environmental water.

Sequential verification of exogenous protein production in OVA gene-targeted chicken bioreactors

The chicken has potential as an efficient bioreactor system because of its outstanding protein production capacity and low cost. The CRISPR/Cas9-mediated gene-editing system enables production of highly marketable exogenous proteins in transgenic chicken bioreactors. However, because it takes approximately 18 mo to evaluate the recombinant protein productivity of the bioreactor due to the generation interval from G0 founders to G1 egg-laying hens, to verification of the exogenous protein at the early stage is difficult. Here we propose a system for sequential validation of exogenous protein production in chicken bioreactors as in hatching female chicks as well as in egg-laying hens. We generated chicken OVALBUMIN (OVA) EGFP knock-in (KI) chicken (OVA EGFP KI) by CRISPR/Cas9-mediated nonhomologous end joining at the chicken OVA gene locus. Subsequently, the estrogen analog, diethylstilbestrol (DES), was subcutaneously implanted in the abdominal region of 1-wk-old OVA EGFP KI female chicks to artificially increase OVALBUMIN expression. The oviducts of DES-treated OVA EGFP KI female chicks expressed OVA and EGFP at the 3-wk-old stage (10 d after DES treatment). We evaluated the expression of EGFP protein in the oviduct, along with the physical properties of eggs and egg white from OVA EGFP KI hens. The rapid identification and isolation of exogenous protein can be confirmed at a very early stage and high-yield production is possible by targeting the chicken oviduct.

Postmenopausal Vaginal and Cervical Cancer Risk Related to In Utero Diethylstilbestrol Exposure.

Women exposed to diethylstilbestrol (DES) in utero have an increased risk of clear cell adenocarcinoma of the lower genital tract, requiring lifelong cervical and vaginal cancer screening. We examined the incidence of DES-related cancers in postmenopausal women 50 years and older. We conducted a retrospective chart review of patients 50 years and older exposed to DES in utero who received care at our institution. Patients were identified using billing codes and/or searching through the electronic record for the word DES. With this 2-pronged approach, we reviewed a total of 503 charts with confirmed DES exposure to identify gynecologic cancer occurrence. Within the 503 selected charts, 28 cases of gynecologic cancer occurrence were identified. Ten patients had cervical cancer and one patient had vaginal cancer. Only 1 woman of 503 developed a DES-related cervical or vaginal malignancy after age 50 years. No patients were diagnosed with cervical or vaginal cancer after age 65 years. Diethylstilbestrol-related malignancies are rare in those older than 50 years. Current cervical cancer screening guidelines recommend cessation of screening in an average risk, adequately screened patient at age 65 years, but patients exposed to DES have historically received lifelong screening. However, we found no cases of cervical or vaginal cancer related to DES after age 65 years, suggesting that screening recommendations could be changed for these patients to align with current screening guidelines.

Development of a photothermal-sensing microfluidic paper-based analytical chip (PT-Chip) for sensitive quantification of diethylstilbestrol

The continuous intake of diethylstilbestrol (DES) residue seriously threatens the health of consumers, so the rapid and sensitive detection of DES is helpful for timely food safety warning. In this study, a photothermal-sensing microfluidic paper-based analytical chip (PT-Chip) was developed for effectively screening of DES residue in foods. Competitive inhibition immunochromatography was introduced and the composite of black phosphorus nanosheets and gold nanoparticles (BP-Au) was used as signal indicators. Then, the test area was irradiated by an 808 nm laser and the photothermal transduction signal was recorded by a thermal imager to achieve sensitive photothermal detection. The amount of DES was obtained by analyzing the temperature change curve of test area. The PT-Chip can achieve accurate quantification of DES with a detection limit of 0.1 μg/L. And the successful determination of DES in different food samples indicating that the PT-Chip can be used as a promising detection device.

Investigation of free and conjugated estrogen fate and emission coefficients in three duck farms.

Concentration animal feeding operation (CAFO) is an important source of environmental estrogen. However, to the best of our knowledge, the data on estrogen discharge during duck breeding and growth is insufficient. This study used liquid chromatography with tandem mass spectrometry (LC/MS/MS) to analyze the free and conjugated estrogen concentrations in the surface water, outlet water, groundwater, and duck manure/soil mixture at three duck farms in Taiwan. Natural estrogen species included estrone (E1), 17β-estradiol (E2), estriol (E3), estrone-3-sulfate (E1-3S), 17β-estradiol-3-sulfate (E2-3S), estrone-3-glucuronide (E1-3G), and 17β-estradiol-3-glucuronide (E2-3G), whereas synthetic estrogen included 17α-ethynylestradiol (EE2) and diethylstilbestrol (DES). This study showed that the total estrogen concentrations in the surface water and groundwater were 15.4 and 4.5ng/L, respectively, which constituted 56% and 58%, respectively, conjugated estrogen. From the pond to the outlet water, the total estrogen concentration decreased by 3.9ng/L (23% loss) in the duck farms. However, the estrogenic potency was slightly reduced from 0.91 to 0.88 E2 equivalent/L, showing a negligible decrease. From the pond to the outlet water, the field results showed that converting the conjugated estrogen into free estrogen in the duck farm-released water increased their environmental hazard. Primarily E1, with an average concentration of 0.9 ± 1.6ng/g, was present in the duck manure. The estrogen excreted by the ducks in the pond (from surface water to outlet water) was estimated to be 0.18kg/million head-year. Although the estrogen concentration in the duck farms was low, the environmental impact of CAFO should not be neglected.

Imine-linked porous covalent organic framework used for the solid-phase extraction of estrogens from honey prior to liquid chromatography-tandem mass spectrometry

以亚胺连接的多孔共价有机骨架材料(IL-COF-1)作为固相萃取的吸附剂,建立了液相色谱-串联质谱快速检测蜂蜜样品中痕量雌激素的方法。该研究选择雌二醇、己烯雌酚、雌三醇、β-雌二醇和炔雌醇5种雌激素作为目标分析物。在蜂蜜样品中添加雌激素,采用单因素优化法对影响萃取效果的重要因素进行优化,获得最佳条件:IL-COF-1用量为30 mg,样品流速为3 mL/min,样品溶液pH值为7,以5 mL的1%(v/v)氨水-甲醇溶液进行洗脱,流速为0.4 mL/min,萃取过程中不添加NaCl。采用高效液相色谱-三重四极杆质谱联用技术对提取物中的雌激素进行定量分析。以乙腈和5 mmol/L的乙酸铵溶液作为流动相进行梯度洗脱,经C18色谱柱分离,采用电喷雾离子源、质谱多反应监测和负离子扫描模式,实现了蜂蜜样品中5种雌激素的快速定性定量分析。在最佳条件下,方法验证结果中雌三醇、β-雌二醇和炔雌醇的线性范围为1~500 ng/g,雌二醇和己烯雌酚的线性范围为0.1~100 ng/g,相关系数(r)为0.9934~0.9972。检出限(S/N=3)为0.01~0.30 ng/g,定量限(S/N=10)为0.05~0.95 ng/g。添加50 ng/g 5种雌激素进行重复性实验,日内精密度相对标准偏差(RSD)为3.2%~6.6%,日间精密度RSD为4.2%~7.9%。基于IL-COF-1的固相萃取-液相色谱-串联质谱法具有快速准确、灵敏度高等特点,适用于蜂蜜中雌激素的分析和检测。将该方法应用于4个实际蜂蜜样品中雌激素的检测,均未检出目标物;在低中高3个水平下,5种雌激素的加标回收率为80.1%~115.2%,结果令人满意。

Evidence Supporting the Diethylstilbestrol Induction of Endometriosis Via Immune-Inflammatory Pathway in Rat Model

Endometriosis is a benign reproductive disorder distinguished by chronic inflammation and anomalous progress of stromal cells and glands outside the uterus. Though the etiology of endometriosis is still a matter of debate, the immuneinflammatory mediators are thought to play a major role. Macrophage infiltration and proinflammatory intermediaries in the peritoneal environment peritoneal milieu impact ovarian, uterine function and pelvic morphology, resulting in endometriosis symptoms and signs. The endometriosis model system was developed using diethylstilbestrol (DES) which is examined to develop the disease via the immune-inflammatory pathway. The immune-inflammatory markers (NFKB, MAPK, TNF-α, IL-6 and STAT- 3) were investigated in this study. The results demonstrated that the DES induced endometriosis animal model had a higher level of CA-125, a biomarker of endometriosis. Endometriosis-induced groups had increased levels of NFKB, MAPK, TNF-α, IL-6 and STAT-3 which clearly depicts the immune-inflammatory role in the induction. Finally, the results conclude that DES induced endometriosis via the immune-inflammatory pathway.&#x0D;