A method was developed for the simultaneous determination of four kinds of estrogens （hexoestrol, diethylstilbestrol, estrone, and 17-beta-estradiol） in feed by gas chromatography-mass spectrometry （GC-MS）. After ...A method was developed for the simultaneous determination of four kinds of estrogens （hexoestrol, diethylstilbestrol, estrone, and 17-beta-estradiol） in feed by gas chromatography-mass spectrometry （GC-MS）. After the sample was extracted by ethyl ether and cleaned-up on HLB phase extraction column, four kinds of estrogens were derived and quantified in gas chromatographymass spectrometry. The results showed that the linear detectable ranged from 2.5 ng· mL-1 to 250 ng· mL-1for hexoestrol and from 5 ng· mL-1 to 500 ng· mL-1 for three other estrogens with the correlation coefficients （R2） were no less than 0.990. The recoveries were in the range of 76.34%-96.33% and the relative standard deviation was no more than 22.7%. The limits of quantitation （LOQ） for all analytics were between 10 ug· kg^-1 and 20 ug· kg^-1. The method was accurate and sensitive and could meet the actual requirements for the analyses of feed samples.展开更多
Objective Determination of estrone (E1) levels has a significant meaning in evaluating physiological effect and diagnosing some diseases. In order to detect free E1 in biological fluids, a monoclonal antibody specifi...Objective Determination of estrone (E1) levels has a significant meaning in evaluating physiological effect and diagnosing some diseases. In order to detect free E1 in biological fluids, a monoclonal antibody specific for E1 was prepared after the complete antigen of E1 was synthesized. The purified monoclonal antibody was fully characterized for later immunoassay. Methods 3-O-carboxymethyl ether derivative of E1 was synthesized and in turn coupled to bovine serum albumin (BSA) to form complete antigen E1-BSA. A monoclonal antibody (McAb) specific for E1 was produced both in vitro and in vivo by a hybridoma anti-E1. Anti-E1 was prepared by fusion of SP2/0 murine myeloma cells with spleen cells isolated from immunized BALB/c mouse. The McAb was characterized by enzyme-linked immunosorbent assay (ELISA), SDS-PAGE and Western-blotting. The specificity of the immunoassay was investigated by determining the cross-reactions of E1 analogs when free E1 was detected by competitive indirect enzyme-linked immunosorbent assay (CI-ELISA). Results Analysis revealed that anti-E1 McAb (E1-McAb) was of the IgG1 type, the molecular weight of E1-McAb was 164 000 daltons. The affinity constant of E1-McAb with coated complete antigen was 8.2108L/mol. The linear range for free E1 determined by CI-ELISA was 10pg/mL^10ng/mL. The detection limit was 21.4 pg/mL (defined as twice the standard deviation of the blank). Conclusion The CI-ELISA developed with E1-McAb was both sensitive and specific. The prepared E1-McAb can be used in some immunoassays.展开更多
Genistein, the main isoflavone from soy, and bisphenol A （BPA）, a food contaminant, are considered ubiquitous xenoestrogens. Here we investigated the influence of genistein and BPA on estrone （El） metabolism in ra...Genistein, the main isoflavone from soy, and bisphenol A （BPA）, a food contaminant, are considered ubiquitous xenoestrogens. Here we investigated the influence of genistein and BPA on estrone （El） metabolism in rat liver microsomes. Both substances inhibited the 2-hydroxylation and 16a-hydroxylation of E1, but in different degrees, thereby reducing the 2-OH-E1/16a-OH-E1 ratio,展开更多
Estrone has been identified as a potential endocrine-disrupting chemical （EDC）[1]. Estrone is usually quantified by gas chromatography-mass spectrometry （GC-MS）, GC-MS/MS, high performance liquid chromatography （...Estrone has been identified as a potential endocrine-disrupting chemical （EDC）[1]. Estrone is usually quantified by gas chromatography-mass spectrometry （GC-MS）, GC-MS/MS, high performance liquid chromatography （HPLC）, HPLC- MS, and HPLC-MS/MS, etc.[2-3]. Meanwhile, several immunoassays based on radioimmunoassay, enzyme linked immunosorbent assay （ELISA） or chemiluminescence immunoassay （CLIA） for determination of estrone in real samples have been developed[2＇4]. Although these methods are sensitive, they need multistage separation and are thus time-consuming and laborious. A very promising way for the simplification of immunoassays for routine applications is a shift from heterogeneous methods （with separation） to homogeneous assays （without separation）[5]. Fluorescence polarization immunoassay （FPIA） is one of the homogeneous techniques that meets the requirements of a simple, reliable, fast, and cost-effective analysis[6]. Therefore, the present study is focused on the development of FPIA in order to analyze estrone based on antibody production.展开更多
A series of estrone derivatives of amino acids and peptides haze been synthesized by different coupling reagents and the binding affinity of deblocked derivatives to the estrogen receptor of rats uteri have been measu...A series of estrone derivatives of amino acids and peptides haze been synthesized by different coupling reagents and the binding affinity of deblocked derivatives to the estrogen receptor of rats uteri have been measured by a comptetive radiometric bind- ing assay.展开更多
Hormone Receptor positive (HR+) breast cancer is the most common malignancy in women. New strategies in the treatments have targeted the estrogen biosynthesis pathways including the inhibition of the aromatase and 17...Hormone Receptor positive (HR+) breast cancer is the most common malignancy in women. New strategies in the treatments have targeted the estrogen biosynthesis pathways including the inhibition of the aromatase and 17β-HSD1 enzymes. The present work, describes the study of a new family of 9 hybrid compounds derived from estrone attached to a coumarin fragment, linked through different lengths of hydrocarbon chains. The activity of these compounds was evaluated by molecular docking with two relevant enzymes in breast cancer (HR+). It has been proposed nine compounds as 17β-HSD1 inhibitors and six as aromatase inhibitors. We found important interactions with key amino acids at the orthosteric site of each enzyme and their score values compared to the crystallographic ligand. The in silico analysis showed good score values in the proposed compounds, where the steroidal portion presented important interactions with Met374 and Tyr155 in aromatase and in 17β-HSD1 respectively. Highlighting Compounds 2, 5 and 8 with an aromatic ring at the C4 position of the coumarin moiety, which favored arene-H type interactions essential for protein-ligand recognition. In addition, the results related to the 17β-HSD1 enzyme demonstrated how the length of the linker influences the interaction;the best score was found for derivative 8 with a chain of 8 methylenes.展开更多
Permanganate was used as an oxidant to control estrone in the present study. Kinetics was determined at pH 2.5-9.4 and temperature 15–40°C for the reaction of estrone with potassium permanganate. It was found th...Permanganate was used as an oxidant to control estrone in the present study. Kinetics was determined at pH 2.5-9.4 and temperature 15–40°C for the reaction of estrone with potassium permanganate. It was found that the reaction is second-order overall and first-order with respect to both estrone and permanganate. The second-order rate constant for the reaction at pH 5.8 and 25°C is 44.45 L mol-1 s-1. The reaction rate first decreased with the increase of pH in the range of 2.5-6.6 and then increased greatly with the increase of pH in the range of 6.6-9.4. In addition, the rate constant exponentially increased with the increase of reaction temperature. Removal of estrogenicity was also investigated during the degradation of estrone using yeast estrogen screen (YES). Results show that the estrogenicity increased in the initial 15 min of reaction and then decreased fast, with a removal rate of 73.8% within the 30 min of reaction. Results also demonstrate that the reaction rate between estrone and permanganate is faster in natural water background than in the ultra-pure water system. Permanganate oxidation is therefore a feasible option for removal of estrone in drinking water treatment processes. However, the contact time must be enough in order to remove estrone without causing the increase of estrogenicity.展开更多
With the continuous developments in labeling techniques, assay methods of radioisotopes and their increasing applications in physiological and biochemical research, it has become both possible and necessary to supply ...With the continuous developments in labeling techniques, assay methods of radioisotopes and their increasing applications in physiological and biochemical research, it has become both possible and necessary to supply highly active labeled estrogens. 6, 7-3H2-estrone and 6, 7-3H2-estradiol are the more widely used commercial products展开更多
Correlations between estrogenic activity and DOC/UV260 ratio in wastewater treatment processes were investigated to propose a simple, reliable and comprehensive indicator for the presence of estrogenic substances. Con...Correlations between estrogenic activity and DOC/UV260 ratio in wastewater treatment processes were investigated to propose a simple, reliable and comprehensive indicator for the presence of estrogenic substances. Contrary to this, when short-term bioassays such as the E-SCREEN, receptor binding and reporter gene expression assays are used for detecting estrogenic activity in the wastewater sample, they require a long time, at least a few days. The major factors contributing to the estrogenic activity were found to be 17β-estradiol （E2） and estrone （El）. A good relationship between the DOC/ UV260 ratio and the concentration of estrogens （El and E2） in the effluent of the activated sludge process was found： the E2 concentration increased as the DOC/UV260 ratio increased while the E1 concentration decreased. The relative estrogenic activity and DOC/UV260 ratio showed a good correlation （R^2 = 0.84） for all sewage samples except the ozonized samples in the sewage treatment plants. This study shows that the estrogenic compounds are hard to be mineralized by the conventional biological processes. Advanced oxidation processes are required to further remove estrogenic substances in the secondary effluent. By analysis of DOC and UV260, the estrogenic activity in the wastewater can be rapidly estimated.展开更多
Natural and synthetic estrogens from sewage treatment systems are suspected to influence the reproductive health of the animals in the rivers. In this article, we investigated the enzymatic treatment of three estroge...Natural and synthetic estrogens from sewage treatment systems are suspected to influence the reproductive health of the animals in the rivers. In this article, we investigated the enzymatic treatment of three estrogens （estrone, 17β-estradiol, and 17α-ethynyletstradiol） by a fungal laccase which oxidize phenolic compounds with dissolved oxygen. The elimination of the estrogenic activities by enzymatic oxidation was demonstrated by medaka vitellogenin assay. In addition, we developed an enzymatic treatment system comprised of β-D-glucuronidase and the laccase for 17β-estradiol 3-（β-D-glucuronide） degradation. The two enzymes eliminated 17β-estradiol 3-（β- D-glucuronide） and the intermediate, 17β-estradiol, efficiently.展开更多
Three series of novel steroidal thiazole derivatives were synthesized by microwave assisted one-potreaction from pregnenolone, testosterone and estrone, respectively. Their structures were characterized by IR, NMRand ...Three series of novel steroidal thiazole derivatives were synthesized by microwave assisted one-potreaction from pregnenolone, testosterone and estrone, respectively. Their structures were characterized by IR, NMRand HRMS, and the antiproliferative activities of all the synthesized compounds against human cervical carcinoma（HeLa）, human liver carcinoma（HepG2）, human lung carcinoma（A549）, nasopharyngeal carcinoma（CNE2） andnormal kidney epithelial cells（HEK293T） were screened. Among the compounds, 1-estron-17＇-ylidene-2-[4＇-（p-bromophenyl）-2＇-thiazol]hydrazone（12） displayed distinct antiproliferative activity against all the tested cancercell lines and was almost inactive to normal kidney epithelial cells.展开更多
Most animals, including humans, produce natural sex hormones such as estrogens: 17β-estradiol(E2) and estrone(E1). These compounds are able to disrupt the reproductive systems of living organisms at trace concentrati...Most animals, including humans, produce natural sex hormones such as estrogens: 17β-estradiol(E2) and estrone(E1). These compounds are able to disrupt the reproductive systems of living organisms at trace concentrations(ng·L^(–1)). This experiment tests the hypothesis that 1% slow pyrolysis biochar-amended sandy soil could retain significant amount of estrogens(E1, E2)from poultry manure in its second year of application. The experiment was conducted over 46 days and consisted of a series of lysimeters containing sandy soil with biocharamended topsoil. The application rate of poultry manure was kept at 2.47 kg·m^(–2). The biochar held a significant concentration of hormone during the first year of its application. However, in the following year(current study), there was no significant retention of hormones in the biochar-amended soil. During the first year after application, the biochar was fresh, so its pores were available for hydrophobic interactions and held significant concentration of hormones. As time passed there were several biotic and abiotic changes on the surface of the biochar so that after some physical fragmentation, pores on the surface were no longer available for hydrophobic interactions. The biochar started releasing dissolved organic carbon, which facilitated greater mobility of hormones from poultry manure down the soil profile.展开更多
基金Supported by Fund of Harbin Provincial Education Department(2014AB3BN041)
文摘A method was developed for the simultaneous determination of four kinds of estrogens （hexoestrol, diethylstilbestrol, estrone, and 17-beta-estradiol） in feed by gas chromatography-mass spectrometry （GC-MS）. After the sample was extracted by ethyl ether and cleaned-up on HLB phase extraction column, four kinds of estrogens were derived and quantified in gas chromatographymass spectrometry. The results showed that the linear detectable ranged from 2.5 ng· mL-1 to 250 ng· mL-1for hexoestrol and from 5 ng· mL-1 to 500 ng· mL-1 for three other estrogens with the correlation coefficients （R2） were no less than 0.990. The recoveries were in the range of 76.34%-96.33% and the relative standard deviation was no more than 22.7%. The limits of quantitation （LOQ） for all analytics were between 10 ug· kg^-1 and 20 ug· kg^-1. The method was accurate and sensitive and could meet the actual requirements for the analyses of feed samples.
基金This work was supported by National Science Foundation of China (Grant no.20075001).
文摘Objective Determination of estrone (E1) levels has a significant meaning in evaluating physiological effect and diagnosing some diseases. In order to detect free E1 in biological fluids, a monoclonal antibody specific for E1 was prepared after the complete antigen of E1 was synthesized. The purified monoclonal antibody was fully characterized for later immunoassay. Methods 3-O-carboxymethyl ether derivative of E1 was synthesized and in turn coupled to bovine serum albumin (BSA) to form complete antigen E1-BSA. A monoclonal antibody (McAb) specific for E1 was produced both in vitro and in vivo by a hybridoma anti-E1. Anti-E1 was prepared by fusion of SP2/0 murine myeloma cells with spleen cells isolated from immunized BALB/c mouse. The McAb was characterized by enzyme-linked immunosorbent assay (ELISA), SDS-PAGE and Western-blotting. The specificity of the immunoassay was investigated by determining the cross-reactions of E1 analogs when free E1 was detected by competitive indirect enzyme-linked immunosorbent assay (CI-ELISA). Results Analysis revealed that anti-E1 McAb (E1-McAb) was of the IgG1 type, the molecular weight of E1-McAb was 164 000 daltons. The affinity constant of E1-McAb with coated complete antigen was 8.2108L/mol. The linear range for free E1 determined by CI-ELISA was 10pg/mL^10ng/mL. The detection limit was 21.4 pg/mL (defined as twice the standard deviation of the blank). Conclusion The CI-ELISA developed with E1-McAb was both sensitive and specific. The prepared E1-McAb can be used in some immunoassays.
基金supported by a POSDRU grantNo.159/1.5/S/136893 grant with title:‘Parteneriat strategic pentru crecterea calitarii cercetarii stiintifice din universitatile medicale prin acordarea de burse doctorale?i postdoctorale-Doc Med.Net_2.0’
文摘Genistein, the main isoflavone from soy, and bisphenol A （BPA）, a food contaminant, are considered ubiquitous xenoestrogens. Here we investigated the influence of genistein and BPA on estrone （El） metabolism in rat liver microsomes. Both substances inhibited the 2-hydroxylation and 16a-hydroxylation of E1, but in different degrees, thereby reducing the 2-OH-E1/16a-OH-E1 ratio,
基金supported by Natural Science Foundation of China(U1301214)Guangdong Natural Science Foundation(S2013030013338)+1 种基金the PhD Programs Foundation of Ministry of Education of China(20114404130002)National Department Public Benefit Research Foundation(201003008-08)
文摘Estrone has been identified as a potential endocrine-disrupting chemical （EDC）[1]. Estrone is usually quantified by gas chromatography-mass spectrometry （GC-MS）, GC-MS/MS, high performance liquid chromatography （HPLC）, HPLC- MS, and HPLC-MS/MS, etc.[2-3]. Meanwhile, several immunoassays based on radioimmunoassay, enzyme linked immunosorbent assay （ELISA） or chemiluminescence immunoassay （CLIA） for determination of estrone in real samples have been developed[2＇4]. Although these methods are sensitive, they need multistage separation and are thus time-consuming and laborious. A very promising way for the simplification of immunoassays for routine applications is a shift from heterogeneous methods （with separation） to homogeneous assays （without separation）[5]. Fluorescence polarization immunoassay （FPIA） is one of the homogeneous techniques that meets the requirements of a simple, reliable, fast, and cost-effective analysis[6]. Therefore, the present study is focused on the development of FPIA in order to analyze estrone based on antibody production.
文摘A series of estrone derivatives of amino acids and peptides haze been synthesized by different coupling reagents and the binding affinity of deblocked derivatives to the estrogen receptor of rats uteri have been measured by a comptetive radiometric bind- ing assay.
文摘Hormone Receptor positive (HR+) breast cancer is the most common malignancy in women. New strategies in the treatments have targeted the estrogen biosynthesis pathways including the inhibition of the aromatase and 17β-HSD1 enzymes. The present work, describes the study of a new family of 9 hybrid compounds derived from estrone attached to a coumarin fragment, linked through different lengths of hydrocarbon chains. The activity of these compounds was evaluated by molecular docking with two relevant enzymes in breast cancer (HR+). It has been proposed nine compounds as 17β-HSD1 inhibitors and six as aromatase inhibitors. We found important interactions with key amino acids at the orthosteric site of each enzyme and their score values compared to the crystallographic ligand. The in silico analysis showed good score values in the proposed compounds, where the steroidal portion presented important interactions with Met374 and Tyr155 in aromatase and in 17β-HSD1 respectively. Highlighting Compounds 2, 5 and 8 with an aromatic ring at the C4 position of the coumarin moiety, which favored arene-H type interactions essential for protein-ligand recognition. In addition, the results related to the 17β-HSD1 enzyme demonstrated how the length of the linker influences the interaction;the best score was found for derivative 8 with a chain of 8 methylenes.
基金supported by the Ministry of Education of China under the scheme of Key Project of Innovation (Grant No. 705013)the National Natural Science Foundation of China under the Scheme of National Creative Research Groups (Grant No. 50821002)
文摘Permanganate was used as an oxidant to control estrone in the present study. Kinetics was determined at pH 2.5-9.4 and temperature 15–40°C for the reaction of estrone with potassium permanganate. It was found that the reaction is second-order overall and first-order with respect to both estrone and permanganate. The second-order rate constant for the reaction at pH 5.8 and 25°C is 44.45 L mol-1 s-1. The reaction rate first decreased with the increase of pH in the range of 2.5-6.6 and then increased greatly with the increase of pH in the range of 6.6-9.4. In addition, the rate constant exponentially increased with the increase of reaction temperature. Removal of estrogenicity was also investigated during the degradation of estrone using yeast estrogen screen (YES). Results show that the estrogenicity increased in the initial 15 min of reaction and then decreased fast, with a removal rate of 73.8% within the 30 min of reaction. Results also demonstrate that the reaction rate between estrone and permanganate is faster in natural water background than in the ultra-pure water system. Permanganate oxidation is therefore a feasible option for removal of estrone in drinking water treatment processes. However, the contact time must be enough in order to remove estrone without causing the increase of estrogenicity.
文摘With the continuous developments in labeling techniques, assay methods of radioisotopes and their increasing applications in physiological and biochemical research, it has become both possible and necessary to supply highly active labeled estrogens. 6, 7-3H2-estrone and 6, 7-3H2-estradiol are the more widely used commercial products
文摘Correlations between estrogenic activity and DOC/UV260 ratio in wastewater treatment processes were investigated to propose a simple, reliable and comprehensive indicator for the presence of estrogenic substances. Contrary to this, when short-term bioassays such as the E-SCREEN, receptor binding and reporter gene expression assays are used for detecting estrogenic activity in the wastewater sample, they require a long time, at least a few days. The major factors contributing to the estrogenic activity were found to be 17β-estradiol （E2） and estrone （El）. A good relationship between the DOC/ UV260 ratio and the concentration of estrogens （El and E2） in the effluent of the activated sludge process was found： the E2 concentration increased as the DOC/UV260 ratio increased while the E1 concentration decreased. The relative estrogenic activity and DOC/UV260 ratio showed a good correlation （R^2 = 0.84） for all sewage samples except the ozonized samples in the sewage treatment plants. This study shows that the estrogenic compounds are hard to be mineralized by the conventional biological processes. Advanced oxidation processes are required to further remove estrogenic substances in the secondary effluent. By analysis of DOC and UV260, the estrogenic activity in the wastewater can be rapidly estimated.
基金supported by the grant from Sasaki Environment Technology Foundation to T.Tanaka
文摘Natural and synthetic estrogens from sewage treatment systems are suspected to influence the reproductive health of the animals in the rivers. In this article, we investigated the enzymatic treatment of three estrogens （estrone, 17β-estradiol, and 17α-ethynyletstradiol） by a fungal laccase which oxidize phenolic compounds with dissolved oxygen. The elimination of the estrogenic activities by enzymatic oxidation was demonstrated by medaka vitellogenin assay. In addition, we developed an enzymatic treatment system comprised of β-D-glucuronidase and the laccase for 17β-estradiol 3-（β-D-glucuronide） degradation. The two enzymes eliminated 17β-estradiol 3-（β- D-glucuronide） and the intermediate, 17β-estradiol, efficiently.
文摘Three series of novel steroidal thiazole derivatives were synthesized by microwave assisted one-potreaction from pregnenolone, testosterone and estrone, respectively. Their structures were characterized by IR, NMRand HRMS, and the antiproliferative activities of all the synthesized compounds against human cervical carcinoma（HeLa）, human liver carcinoma（HepG2）, human lung carcinoma（A549）, nasopharyngeal carcinoma（CNE2） andnormal kidney epithelial cells（HEK293T） were screened. Among the compounds, 1-estron-17＇-ylidene-2-[4＇-（p-bromophenyl）-2＇-thiazol]hydrazone（12） displayed distinct antiproliferative activity against all the tested cancercell lines and was almost inactive to normal kidney epithelial cells.
基金support provided by the Natural Sciences and Engineering Research Council of Canada (NSERC) under the project Reducing Environmental Pollution from Veterinary Antibiotics, Endocrine Disruptive Chemicals, and Pathogens in Livestock Manurethe Department of Bioresource Engineering, McGill University for providing a Liliane and David Stewart Fellowship in Water Resources for support of this research work
文摘Most animals, including humans, produce natural sex hormones such as estrogens: 17β-estradiol(E2) and estrone(E1). These compounds are able to disrupt the reproductive systems of living organisms at trace concentrations(ng·L^(–1)). This experiment tests the hypothesis that 1% slow pyrolysis biochar-amended sandy soil could retain significant amount of estrogens(E1, E2)from poultry manure in its second year of application. The experiment was conducted over 46 days and consisted of a series of lysimeters containing sandy soil with biocharamended topsoil. The application rate of poultry manure was kept at 2.47 kg·m^(–2). The biochar held a significant concentration of hormone during the first year of its application. However, in the following year(current study), there was no significant retention of hormones in the biochar-amended soil. During the first year after application, the biochar was fresh, so its pores were available for hydrophobic interactions and held significant concentration of hormones. As time passed there were several biotic and abiotic changes on the surface of the biochar so that after some physical fragmentation, pores on the surface were no longer available for hydrophobic interactions. The biochar started releasing dissolved organic carbon, which facilitated greater mobility of hormones from poultry manure down the soil profile.
