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[PMID]:28870733
[Au] Autor:Stenger B; Gerber A; Bernhardt R; Hannemann F
[Ad] Endereço:Institute of Biochemistry, Saarland University, Saarbrücken, Germany.
[Ti] Título:Functionalized poly(3-hydroxybutyric acid) bodies as new in vitro biocatalysts.
[So] Source:Biochim Biophys Acta;1866(1):52-59, 2018 01.
[Is] ISSN:0006-3002
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Cytochromes P450 play a key role in the drug and steroid metabolism in the human body. This leads to a high interest in this class of proteins. Mammalian cytochromes P450 are rather delicate. Due to their localization in the mitochondrial or microsomal membrane, they tend to aggregate during expression and purification and to convert to an inactive form so that they have to be purified and stored in complex buffers. The complex buffers and low storage temperatures, however, limit the feasibility of fast, automated screening of the corresponding cytochrome P450-effector interactions, which are necessary to study substrate-protein and inhibitor-protein interactions. Here, we present the production and isolation of functionalized poly(3-hydroxybutyrate) granules (PHB bodies) from Bacillus megaterium MS941 strain. In contrast to the expression in Escherichia coli, where mammalian cytochromes P450 are associated to the cell membrane, when CYP11A1 is heterologously expressed in Bacillus megaterium, it is located on the PHB bodies. The surface of these particles provides a matrix for immobilization and stabilization of the CYP11A1 during the storage of the protein and substrate conversion. It was demonstrated that the PHB polymer basis is inert concerning the performed conversion. Immobilization of the CYP11A1 onto the PHB bodies allows freeze-drying of the complex without significant decrease of the CYP11A1 activity. This is the first lyophilization of a mammalian cytochrome P450, which allows storage over more than 18days at 4°C instead of storage at -80°C. In addition, we were able to immobilize the cytochrome P450 on the PHB bodies in vitro. In this case the expression of the protein is separated from the production of the immobilization matrix, which widens the application of this method. This article is part of a Special Issue entitled: Cytochrome P450 biodiversity and biotechnology, edited by Erika Plettner, Gianfranco Gilardi, Luet Wong, Vlada Urlacher, Jared Goldstone.
[Mh] Termos MeSH primário: Ácido 3-Hidroxibutírico/química
Bacillus megaterium/genética
Biotecnologia/métodos
Enzima de Clivagem da Cadeia Lateral do Colesterol/química
Proteínas Imobilizadas/biossíntese
Proteínas Mitocondriais/biossíntese
[Mh] Termos MeSH secundário: Ácido 3-Hidroxibutírico/biossíntese
Animais
Bacillus megaterium/enzimologia
Biocatálise
Bovinos
Colesterol/química
Colesterol/metabolismo
Enzima de Clivagem da Cadeia Lateral do Colesterol/genética
Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo
Grânulos Citoplasmáticos/química
Liofilização
Expressão Gênica
Proteínas Imobilizadas/química
Proteínas Imobilizadas/genética
Proteínas Mitocondriais/química
Proteínas Mitocondriais/genética
Pregnenolona/biossíntese
Pregnenolona/química
Refrigeração
Transgenes
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Immobilized Proteins); 0 (Mitochondrial Proteins); 73R90F7MQ8 (Pregnenolone); 97C5T2UQ7J (Cholesterol); EC 1.14.15.6 (Cholesterol Side-Chain Cleavage Enzyme); TZP1275679 (3-Hydroxybutyric Acid)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180208
[Lr] Data última revisão:
180208
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170906
[St] Status:MEDLINE


  2 / 1832 MEDLINE  
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[PMID]:29178636
[Au] Autor:Lara-Velazquez M; Perdomo-Pantoja A; Blackburn PR; Gass JM; Caulfield TR; Atwal PS
[Ad] Endereço:Department of Neurosurgery, Mayo Clinic, Jacksonville, Florida.
[Ti] Título:A novel splice site variant in CYP11A1 in trans with the p.E314K variant in a male patient with congenital adrenal insufficiency.
[So] Source:Mol Genet Genomic Med;5(6):781-787, 2017 11.
[Is] ISSN:2324-9269
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: The CYP11A1 gene encodes the cytochrome P450 side-chain cleavage enzyme, which is essential for steroid formation. Recessive variants in this gene can lead to impairment of sexual differentiation caused by a complete or partial loss of steroid hormone production. The phenotypic spectrum in affected 46XY males may vary from surgically repairable defects including cryptorchidism and hypospadias to complete feminization of external gonads, accompanied by symptoms of adrenal dysfunction. METHODS: Whole-exome sequencing (WES) of a 12-year-old male proband and his parents was performed after a protracted diagnostic odyssey failed to uncover the cause of his primary adrenal insufficiency. Of note, the proband had early symptomatology and corrective surgery for hypospadias, raising suspicion for a disorder of steroidogenesis. RESULTS: WES identified compound heterozygous variants in CYP11A1 including a novel canonical splice site variant (c.425+1G>A) and a previously reported p.E314K variant, which were consistent with a diagnosis of congenital adrenal insufficiency with partial 46XY sex reversal. CONCLUSION: Congenital adrenal insufficiency with 46XY sex reversal is a rare disorder that is characterized by dysregulation of steroid hormone synthesis, leading to adrenal and gonadal dysfunction. In this report, we describe a patient with adrenal insufficiency, hypospadias, and skin hyperpigmentation who was found to have a novel c.425+1G>A variant in trans with the p.E314K variant in CYP11A1. We performed structural analyses to examine the effect of the p.E314K variant on protein function and show that it falls in the core of the protein may disrupt cholesterol binding in the active site.
[Mh] Termos MeSH primário: Insuficiência Adrenal/congênito
Enzima de Clivagem da Cadeia Lateral do Colesterol/genética
[Mh] Termos MeSH secundário: Insuficiência Adrenal/diagnóstico
Insuficiência Adrenal/genética
Criança
Enzima de Clivagem da Cadeia Lateral do Colesterol/química
Análise Mutacional de DNA
Heterozigoto
Seres Humanos
Masculino
Linhagem
Fenótipo
Polimorfismo de Nucleotídeo Único
Estrutura Terciária de Proteína
Sítios de Splice de RNA/genética
Sequenciamento Completo do Exoma
[Pt] Tipo de publicação:CASE REPORTS; JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (RNA Splice Sites); EC 1.14.15.6 (Cholesterol Side-Chain Cleavage Enzyme)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180207
[Lr] Data última revisão:
180207
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171128
[St] Status:MEDLINE
[do] DOI:10.1002/mgg3.322


  3 / 1832 MEDLINE  
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[PMID]:28743557
[Au] Autor:Zhao JL; Zhao YY; Zhu WJ
[Ad] Endereço:Department of Developmental and Regenerative Biology, College of Life Science and Technology, Jinan University, Guangdong, Guangzhou 510632, China.
[Ti] Título:A high-fat, high-protein diet attenuates the negative impact of casein-induced chronic inflammation on testicular steroidogenesis and sperm parameters in adult mice.
[So] Source:Gen Comp Endocrinol;252:48-59, 2017 Oct 01.
[Is] ISSN:1095-6840
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The interaction between obesity and chronic inflammation has been studied. Diet-induced obesity or chronic inflammation could reduce the testicular functions of males. However, the mechanism underlying the reproductive effects of fattening foods in males with or without chronic inflammation still needs further discussion. This study was aimed to investigate the effects of high-fat, high-protein diet on testicular steroidogenesis and sperm parameters in adult mice under physiological and chronic inflammatory conditions. Because casein can trigger a non-infectious systemic inflammatory response, we used casein injection to induce chronic inflammation in male adult Kunming mice. Twenty-four mice were randomly and equally divided into four groups: (i) normal diet+saline (Control); (ii) normal diet+casein (ND+CS); (iii) high-fat, high-protein diet+saline (HFPD+SI); (iv) high-fat, high-protein diet+casein (HFPD+CS). After 8weeks, there was a significant increase in body weight for groups HFPD+SI and HFPD+CS and a decrease in group ND+CS compared with the control. The serum levels of tumor necrosis factor alpha (TNF-α), interleukin-10 (IL-10) and lipid profiles were increased markedly in groups ND+CS, HFPD+SI and HFPD+CS compared with the control. A remarkable reduction of serum adiponectin level occurred in group HFPD+CS compared with group ND+CS. Sperm parameters (sperm count, viability and abnormality) were also adversely affected in groups ND+CS and HFPD+SI. Groups ND+CS and HFPD+SI showed severe pathological changes in testicular tissues. Semiquantitative RT-PCR, Western blot and immunohistochemical staining also showed significant reductions in both testicular mRNA and protein levels of steroidogenic acute regulatory (StAR) and cytochrome P450scc (CYP11A1) in groups HFPD+SI and HFPD+CS compared with the control, whereas testicular mRNA and protein levels of 3ß-hydroxysteroid dehydrogenase (3ß-HSD) in groups HFPD+SI and HFPD+CS significantly increased. The mRNA and protein levels of the StAR and 3ß-HSD in group HFPD+CS were both higher than those of in group ND+CS. These results indicated that Kunming male mice with high-fat, high-protein diet and casein injection for 8weeks can be used to establish a diet-induced obesity and chronic systemic inflammation. The sperm parameters in groups ND+CS and HFPD+SI decreased accompanied by pathological changes of testicular tissue. This resultant effect of reduced serum testosterone levels was associated with the overproduction of TNF-α and IL-10 and down-regulation of StAR and CYP11A1. Under the same casein-induced chronic inflammation condition, the mice with high-fat, high-protein diet had better testicular steroidogenesis activity and sperm parameters compared with the mice in normal diet, indicating that the mice with casein-induced inflammatory injury consuming a high-fat, high-protein diet gained weight normally, reduced serum adiponectin level and increased testosterone production by an upregulation of 3ß-HSD expression. High-fat, high-protein diet attenuated the negative impact of casein-induced chronic inflammation on testicular steroidogenesis and sperm parameters.
[Mh] Termos MeSH primário: Caseínas/toxicidade
Dieta Hiperlipídica
Dieta Rica em Proteínas
Inflamação/patologia
Espermatozoides/metabolismo
Esteroides/biossíntese
Testículo/metabolismo
[Mh] Termos MeSH secundário: Animais
Peso Corporal/efeitos dos fármacos
Forma Celular/efeitos dos fármacos
Sobrevivência Celular/efeitos dos fármacos
Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo
Citocinas/metabolismo
Inflamação/metabolismo
Mediadores da Inflamação/metabolismo
Lipídeos/sangue
Masculino
Camundongos
Contagem de Espermatozoides
Testículo/efeitos dos fármacos
Testosterona/sangue
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Caseins); 0 (Cytokines); 0 (Inflammation Mediators); 0 (Lipids); 0 (Steroids); 3XMK78S47O (Testosterone); EC 1.14.15.6 (Cholesterol Side-Chain Cleavage Enzyme)
[Em] Mês de entrada:1712
[Cu] Atualização por classe:171222
[Lr] Data última revisão:
171222
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170727
[St] Status:MEDLINE


  4 / 1832 MEDLINE  
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[PMID]:29189770
[Au] Autor:Pechkova E; Nicolini C
[Ad] Endereço:Laboratories of Biophysics and Nanotechnology, University of Genoa Medical School, Genoa, Italy.
[Ti] Título:Langmuir-Blodgett nanotemplates for protein crystallography.
[So] Source:Nat Protoc;12(12):2570-2589, 2017 Dec.
[Is] ISSN:1750-2799
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The new generation of synchrotrons and microfocused beamlines has enabled great progress in X-ray protein crystallography, resulting in new 3D atomic structures for proteins of high interest to the pharmaceutical industry and life sciences. It is, however, often still challenging to produce protein crystals of sufficient size and quality (order, intensity of diffraction, radiation stability). In this protocol, we provide instructions for performing the Langmuir-Blodgett (LB) nanotemplate method, a crystallization approach that can be used for any protein (including membrane proteins). We describe how to produce highly ordered 2D LB protein monolayers at the air-water interface and deposit them on glass slides. LB-film formation can be observed by surface-pressure measurements and Brewster angle microscopy (BAM), although its quality can be characterized by atomic force microscopy (AFM) and nanogravimetry. Such films are then used as a 2D template for triggering 3D protein crystal formation by hanging-drop vapor diffusion. The procedure for forming the 2D template takes a few minutes. Structural information about the protein reorganization in the LB film during the crystallization process on the nano level can be obtained using an in situ submicron GISAXS (grazing-incidence small-angle X-ray scattering) method. MicroGISAXS spectra, measured directly at the interface of the LB films and protein solution in real time, as described in this protocol, can be interpreted in terms of the buildup of layers, islands, or holes. In our experience, the obtained LB crystals take 1-10 d to prepare and they are more ordered and radiation stable as compared with those produced using other crystallization methods.
[Mh] Termos MeSH primário: Cristalização/métodos
Cristalografia por Raios X/métodos
Nanoestruturas/química
Proteínas/química
[Mh] Termos MeSH secundário: Animais
Bovinos
Galinhas
Enzima de Clivagem da Cadeia Lateral do Colesterol/química
Cristalização/instrumentação
Cristalografia por Raios X/instrumentação
Desenho de Equipamento
Marantaceae/química
Modelos Moleculares
Muramidase/química
Proteínas de Plantas/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Plant Proteins); 0 (Proteins); 53850-34-3 (thaumatin protein, plant); EC 1.14.15.6 (Cholesterol Side-Chain Cleavage Enzyme); EC 3.2.1.17 (Muramidase)
[Em] Mês de entrada:1712
[Cu] Atualização por classe:171219
[Lr] Data última revisão:
171219
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171201
[St] Status:MEDLINE
[do] DOI:10.1038/nprot.2017.108


  5 / 1832 MEDLINE  
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[PMID]:28991453
[Au] Autor:Zhu Q; Mak PJ; Tuckey RC; Kincaid JR
[Ad] Endereço:Department of Chemistry, Marquette University , Milwaukee, Wisconsin 53233, United States.
[Ti] Título:Active Site Structures of CYP11A1 in the Presence of Its Physiological Substrates and Alterations upon Binding of Adrenodoxin.
[So] Source:Biochemistry;56(43):5786-5797, 2017 Oct 31.
[Is] ISSN:1520-4995
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The rate-limiting step in the steroid synthesis pathway is catalyzed by CYP11A1 through three sequential reactions. The first two steps involve hydroxylations at positions 22 and 20, generating 20(R),22(R)-dihydroxycholesterol (20R,22R-DiOHCH), with the third stage leading to a C20-C22 bond cleavage, forming pregnenolone. This work provides detailed information about the active site structure of CYP11A1 in the resting state and substrate-bound ferric forms as well as the CO-ligated adducts. In addition, high-quality resonance Raman spectra are reported for the dioxygen complexes, providing new insight into the status of Fe-O-O fragments encountered during the enzymatic cycle. Results show that the three natural substrates of CYP11A1 have quite different effects on the active site structure, including variations of spin state populations, reorientations of heme peripheral groups, and, most importantly, substrate-mediated distortions of Fe-CO and Fe-O fragments, as revealed by telltale shifts of the observed vibrational modes. Specifically, the vibrational mode patterns observed for the Fe-O-O fragments with the first and third substrates are consistent with H-bonding interactions with the terminal oxygen, a structural feature that tends to promote O-O bond cleavage to form the Compound I intermediate. Furthermore, such spectral data are acquired for complexes with the natural redox partner, adrenodoxin (Adx), revealing protein-protein-induced active site structural perturbations. While this work shows that Adx has an only weak effect on ferric and ferrous CO states, it has a relatively stronger impact on the Fe-O-O fragments of the functionally relevant oxy complexes.
[Mh] Termos MeSH primário: Adrenodoxina/química
Enzima de Clivagem da Cadeia Lateral do Colesterol/química
Modelos Moleculares
[Mh] Termos MeSH secundário: Adrenodoxina/metabolismo
Domínio Catalítico
Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo
Seres Humanos
Estrutura Quaternária de Proteína
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
12687-22-8 (Adrenodoxin); EC 1.14.15.6 (Cholesterol Side-Chain Cleavage Enzyme)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171103
[Lr] Data última revisão:
171103
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171010
[St] Status:MEDLINE
[do] DOI:10.1021/acs.biochem.7b00766


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[PMID]:28964809
[Au] Autor:Wang W; Sun Y; Guo Y; Cai P; Li Y; Liu J; Cai G; Kiyoshi A; Zhang W
[Ad] Endereço:Department of Health Inspection and Quarantine, School of Public Health, Fujian Medical University, Fuzhou, Fujian, China; Fujian Province Key Laboratory of Environment and Health, School of Public Health, Fujian Medical University, Fuzhou, Fujian, China. Electronic address: wangwenxiang@fjmu.edu.cn
[Ti] Título:Continuous soy isoflavones exposure from weaning to maturity induces downregulation of ovarian steroidogenic factor 1 gene expression and corresponding changes in DNA methylation pattern.
[So] Source:Toxicol Lett;281:175-183, 2017 Nov 05.
[Is] ISSN:1879-3169
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Female Wistar rats were treated with orally administered soy isoflavones at concentrations of 0, 25, 50, or 100mg/kg body weight from weaning until sexual maturity (3 mo.), and ovarian steroidogenesis was evaluated. After soy isoflavones were administered, a significant (P<0.05) decrease (44%) in the serum estrodial levels of the high-dose (HD) group were observed. Cultured granulosa cells from the middle- (MD) and HD groups showed significantly (P<0.05) reduced (31%, 45%, respectively) in vitro estradiol secretion, and those from the HD group showed significantly (P<0.05) reduced progesterone (25%) secretion. Compared with the control group, the mRNA expression of the steroidogenic acute regulatory protein (Star), cytochromeP450 cholesterol side chain cleavage (Cyp11a1 and Cyp19a1), and hydroxysteroid dehydrogenase 3b (Hsd3b) genes also decreased. Real-time quantitative PCR and Western blotting revealed a significant (P<0.05) decrease in key transcription factor steroidogenic factor-1 (SF-1) expression in the HD group. The detection of DNA methylation using bisulfitesequencing PCR (BSP) suggested a significantly (P<0.05) increased total methylation rate in the proximal SF-1 promoter in the HD group. Further studies showed that treatment with soy isoflavones can significantly (P<0.05) increase the mRNA expression of DNA methyltransferase (DNMT) 1 and DNMT3a. This study proved that soy isoflavone administration from weaning until sexual maturity could inhibit ovarian steroidogenesis, suggesting that SF-1 might play an important role in this effect. In addition, DNA methylation might play a role in the downregulation of SF-1 gene expression induced by soy isoflavones.
[Mh] Termos MeSH primário: Metilação de DNA/efeitos dos fármacos
Isoflavonas/farmacologia
Fator Esteroidogênico 1/metabolismo
Desmame
[Mh] Termos MeSH secundário: Animais
Aromatase/genética
Aromatase/metabolismo
Enzima de Clivagem da Cadeia Lateral do Colesterol/genética
Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo
DNA (Citosina-5-)-Metiltransferase 1
DNA (Citosina-5-)-Metiltransferases/genética
DNA (Citosina-5-)-Metiltransferases/metabolismo
Relação Dose-Resposta a Droga
Regulação para Baixo
Estradiol/sangue
Feminino
Ovário/efeitos dos fármacos
Ovário/metabolismo
Fosfoproteínas/genética
Fosfoproteínas/metabolismo
Regiões Promotoras Genéticas
Ratos
Ratos Wistar
Feijão de Soja/química
Fator Esteroidogênico 1/genética
Fatores de Transcrição/genética
Fatores de Transcrição/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Isoflavones); 0 (Phosphoproteins); 0 (Steroidogenic Factor 1); 0 (Transcription Factors); 0 (steroidogenic acute regulatory protein); 0 (steroidogenic factor 1, rat); 4TI98Z838E (Estradiol); EC 1.14.14.1 (Aromatase); EC 1.14.15.6 (Cholesterol Side-Chain Cleavage Enzyme); EC 2.1.1.37 (DNA (Cytosine-5-)-Methyltransferase 1); EC 2.1.1.37 (DNA (Cytosine-5-)-Methyltransferases); EC 2.1.1.37 (DNA methyltransferase 3A)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171002
[St] Status:MEDLINE


  7 / 1832 MEDLINE  
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[PMID]:28501544
[Au] Autor:Zhang T; Liu Y; Chen H; Gao J; Zhang Y; Yuan C; Wang Z
[Ad] Endereço:College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, China.
[Ti] Título:The DNA methylation status alteration of two steroidogenic genes in gonads of rare minnow after bisphenol A exposure.
[So] Source:Comp Biochem Physiol C Toxicol Pharmacol;198:9-18, 2017 Aug.
[Is] ISSN:1532-0456
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Both cytochrome P450c17 (CYP17A1) and P-450 side chain cleavage (CYP11A1) play important roles in steroid biosynthesis. According to our previous studies, bisphenol A (BPA) could regulate the mRNA expression of cyp17a1 and cyp11a1 in rare minnow Gobiocypris rarus. However, the potential mechanism of the regulation is barely understood. In the present study, aiming to explore how BPA affects the mRNA expression of cyp17a1 and cyp11a1 in testes and ovaries of G. rarus, we firstly cloned 340-bp fragment of 5' flanking region of cyp11a1 and then detected the methylation level of CpG loci involved in 5' flanking of cyp11a1 and cyp17a1 and their mRNA expression levels. Results showed that exposure to BPA significantly increased serum estradiol (E2) and 11-ketotesterone (11-KT) concentrations. Ovarian mRNA expression of cyp17a1 and cyp11a1 were significantly decreased after BPA exposure 7- for and 14-days. However, transcriptions of testicular cyp17a1 and cyp11a1 were significantly increased and decreased respectively after BPA treatment for 14days. The DNA methylation levels of cyp17a1 were decreased in ovaries on day 7 and increased in ovaries and decreased in testes respectively on day 14. The methylation levels of cyp11a1 were increased in ovaries on day 7 and both ovaries and testes on day 14. There were a significant correlation between DNA methylation at specific CpG loci and cyp17a1 and cyp11a1 genes transcription levels. In conclusion, the CpG loci methylation in 5' flanking region appears to involve in the regulation of mRNA expression of cyp17a1 and cyp11a1 mediated by BPA.
[Mh] Termos MeSH primário: Compostos Benzidrílicos/toxicidade
Enzima de Clivagem da Cadeia Lateral do Colesterol/genética
Metilação de DNA/efeitos dos fármacos
Fenóis/toxicidade
Esteroide 17-alfa-Hidroxilase/genética
[Mh] Termos MeSH secundário: Regiões 5' não Traduzidas
Animais
Peso Corporal/efeitos dos fármacos
Clonagem Molecular
Cyprinidae
Ecotoxicologia/métodos
Estradiol/sangue
Feminino
Proteínas de Peixes/genética
Regulação da Expressão Gênica/efeitos dos fármacos
Masculino
Ovário/efeitos dos fármacos
Ovário/fisiologia
Testículo/efeitos dos fármacos
Testículo/fisiologia
Testosterona/análogos & derivados
Testosterona/sangue
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (5' Untranslated Regions); 0 (Benzhydryl Compounds); 0 (Fish Proteins); 0 (Phenols); 3XMK78S47O (Testosterone); 4TI98Z838E (Estradiol); EC 1.14.14.19 (Steroid 17-alpha-Hydroxylase); EC 1.14.15.6 (Cholesterol Side-Chain Cleavage Enzyme); KF38W1A85U (11-ketotestosterone); MLT3645I99 (bisphenol A)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170728
[Lr] Data última revisão:
170728
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170515
[St] Status:MEDLINE


  8 / 1832 MEDLINE  
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[PMID]:28414290
[Au] Autor:Ershov PV; Yablokov EO; Mezentsev YV; Kalushskiy LA; Florinskaya AV; Veselovsky AV; Gnedenko OV; Gilep AA; Usanov SA; Medvedev AE; Ivanov AS
[Ad] Endereço:Institute of Biomedical Chemistry, Moscow, Russia.
[Ti] Título:[The effect of isatin on protein-protein interactions between cytochrome b5 and cytochromes P450].
[Ti] Título:Vliianie izatina na vzaimodeistvie tsitokhroma b5 s tsitokhromami R450..
[So] Source:Biomed Khim;63(2):170-175, 2017 Mar.
[Is] ISSN:2310-6972
[Cp] País de publicação:Russia (Federation)
[La] Idioma:rus
[Ab] Resumo:Cytochromes P450 (CYP) are involved in numerous biochemical processes including metabolism of xenobiotics, biosynthesis of cholesterol, steroid hormones etc. Since some CYP catalyze indol oxidation to isatin, we have hypothesized that isatin can regulate protein-protein interactions (PPI) between components of the CYP system thus representing a (negative?) feedback mechanism. The aim of this study was to investigate a possible effect of isatin on interaction of human CYP with cytochrome b5 (CYB5A). Using the optical biosensor test system employing surface plasmon resonance (SPR) we have investigated interaction of immobilized CYB5A with various CYP in the absence and in the presence of isatin. The SPR-based experiments have shown that a high concentration of isatin (270 mM) increases Kd values for complexes CYB5A/CYP3А5 and CYB5A/CYP3A4 (twofold and threefold, respectively), but has no influence on complex formation between CYB5A and other CYP (including indol-metabolizing CYP2C19 and CYP2E1). Isatin injection to the optical biosensor chip with the preformed molecular complex CYB5A/CYP3A4 caused a 30%-increase in its dissociation rate. Molecular docking manipulations have shown that isatin can influence interaction of CYP3А5 or CYP3A4 with CYB5A acting at the contact region of CYB5A/CYP.
[Mh] Termos MeSH primário: Citocromo P-450 CYP2C19/química
Citocromo P-450 CYP2E1/química
Citocromo P-450 CYP3A/química
Citocromos b5/química
Isatina/química
[Mh] Termos MeSH secundário: Sítios de Ligação
Enzima de Clivagem da Cadeia Lateral do Colesterol/química
Citocromo P-450 CYP2C9/química
Seres Humanos
Cinética
Simulação de Acoplamento Molecular
Ligação Proteica
Conformação Proteica em alfa-Hélice
Conformação Proteica em Folha beta
Domínios e Motivos de Interação entre Proteínas
Proteínas Recombinantes/química
Soluções
Esteroide 11-beta-Hidroxilase/química
Esteroide 17-alfa-Hidroxilase/química
Ressonância de Plasmônio de Superfície
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Recombinant Proteins); 0 (Solutions); 82X95S7M06 (Isatin); 9035-39-6 (Cytochromes b5); EC 1.14.13.- (CYP2C9 protein, human); EC 1.14.13.- (Cytochrome P-450 CYP2C9); EC 1.14.13.- (Cytochrome P-450 CYP2E1); EC 1.14.13.67 (CYP3A4 protein, human); EC 1.14.14.1 (CYP2C19 protein, human); EC 1.14.14.1 (CYP3A5 protein, human); EC 1.14.14.1 (Cytochrome P-450 CYP2C19); EC 1.14.14.1 (Cytochrome P-450 CYP3A); EC 1.14.14.19 (CYP17A1 protein, human); EC 1.14.14.19 (Steroid 17-alpha-Hydroxylase); EC 1.14.15.4 (Steroid 11-beta-Hydroxylase); EC 1.14.15.6 (Cholesterol Side-Chain Cleavage Enzyme)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170912
[Lr] Data última revisão:
170912
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170418
[St] Status:MEDLINE
[do] DOI:10.18097/PBMC20176302170


  9 / 1832 MEDLINE  
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[PMID]:28381288
[Au] Autor:Adibi JJ; Buckley JP; Lee MK; Williams PL; Just AC; Zhao Y; Bhat HK; Whyatt RM
[Ad] Endereço:Department of Epidemiology, Graduate School of Public Health, University of Pittsburgh, 130 Desoto Street, Parran Hall 5132, Pittsburgh, PA, 15261, USA. adibij@pitt.edu.
[Ti] Título:Maternal urinary phthalates and sex-specific placental mRNA levels in an urban birth cohort.
[So] Source:Environ Health;16(1):35, 2017 Apr 05.
[Is] ISSN:1476-069X
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Prenatal urinary concentrations of phthalates in women participants in an urban birth cohort were associated with outcomes in their children related to neurodevelopment, autoimmune disease risk, and fat mass at 3,5,7, and 8 years of life. Placental biomarkers and outcomes at birth may offer biologic insight into these associations. This is the first study to address these associations with candidate genes from the phthalate and placenta literature, accounting for sex differences, and using absolute quantitation methods for mRNA levels. METHODS: We measured candidate mRNAs in 180 placentas sampled at birth (HSD17B1, AHR, CGA, CYP19A1, SLC27A4, PTGS2, PPARG, CYP11A1) by quantitative PCR and an absolute standard curve. We estimated associations of log mRNA with quartiles of urinary phthalate monoesters using linear mixed models. Phthalate metabolites (N = 358) and mRNAs (N = 180) were transformed to a z-score and modeled as independent, correlated vectors in relation to large for gestational age (LGA) and gestational diabetes mellitus (GDM). RESULTS: CGA was associated with 4 out of 6 urinary phthalates. CGA was 2.0 log units lower at the 3 vs. 1 quartile of mono-n-butyl phthalate (MnBP) (95% confidence interval (CI): -3.5, -0.5) in male placentas, but 0.6 log units higher (95% CI: -0.8, 1.9) in female placentas (sex interaction p = 0.01). There was an inverse association of MnBP with PPARG in male placentas (-1.1 log units at highest vs. lowest quartile, 95% CI: -2.0, -0.1). CY19A1, CYP11A1, CGA were associated with one or more of the following in a sex-specific manner: monobenzyl phthalate (MBzP), MnBP, mono-iso-butyl phthalate (MiBP). These 3 mRNAs were lower by 1.4-fold (95% CI: -2.4, -1.0) in male GDM placentas vs. female and non-GDM placentas (p-value for interaction = 0.04). The metabolites MnBP/MiBP were 16% higher (95% CI: 0, 22) in GDM pregnancies. CONCLUSIONS: Prenatal concentrations of certain phthalates and outcomes at birth were modestly associated with molecular changes in fetal placental tissue during pregnancy. Associations were stronger in male vs. female placentas, and associations with MnBP and MiBP were stronger than other metabolites. Placental mRNAs are being pursued further as potential mediators of exposure-induced risks to the health of the child.
[Mh] Termos MeSH primário: Poluentes Ambientais/urina
Exposição Materna
Ácidos Ftálicos/urina
Placenta/metabolismo
RNA Mensageiro/metabolismo
[Mh] Termos MeSH secundário: Adulto
Aromatase/genética
Enzima de Clivagem da Cadeia Lateral do Colesterol/genética
Estradiol Desidrogenases/genética
Proteínas de Transporte de Ácido Graxo/genética
Feminino
Expressão Gênica
Subunidade alfa de Hormônios Glicoproteicos/genética
Seres Humanos
Masculino
PPAR gama/genética
Gravidez
Receptores de Hidrocarboneto Arílico/genética
Caracteres Sexuais
População Urbana
Adulto Jovem
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Environmental Pollutants); 0 (Fatty Acid Transport Proteins); 0 (Glycoprotein Hormones, alpha Subunit); 0 (PPAR gamma); 0 (Phthalic Acids); 0 (RNA, Messenger); 0 (Receptors, Aryl Hydrocarbon); 0 (SLC27A4 protein, human); EC 1.1.1.62 (Estradiol Dehydrogenases); EC 1.1.1.62 (HSD17B1 protein, human); EC 1.14.14.1 (Aromatase); EC 1.14.14.1 (CYP19A1 protein, human); EC 1.14.15.6 (Cholesterol Side-Chain Cleavage Enzyme)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171003
[Lr] Data última revisão:
171003
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170407
[St] Status:MEDLINE
[do] DOI:10.1186/s12940-017-0241-5


  10 / 1832 MEDLINE  
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[PMID]:28302174
[Au] Autor:Li J; Zhou Q; Ma Z; Wang M; Shen WJ; Azhar S; Guo Z; Hu Z
[Ad] Endereço:Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Sciences, Nanjing Normal University, 1 WenYuan Road, Nanjing, 210023, China.
[Ti] Título:Feedback inhibition of CREB signaling by p38 MAPK contributes to the negative regulation of steroidogenesis.
[So] Source:Reprod Biol Endocrinol;15(1):19, 2017 Mar 16.
[Is] ISSN:1477-7827
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Steroidogenesis is a complex, multi-steps biological process in which, cholesterol precursor is converted to steroids in a tissue specific and tropic hormone dependent manner. Given that steroidogenesis is achieved by coordinated functioning of multiple tissue specific enzymes, many steroids intermediates/metabolites are generated during this process. Both the steroid products as well as major lipoprotein cholesterol donor, high-density lipoprotein 3 (hHDL ) have the potential to negatively regulate steroidogenesis via increased oxidative stress/reactive oxygen species (ROS) generation. METHODS: In the current study, we examined the effects of treatment of a mouse model of steroidogenesis, Y1-BS1 adrenocortical tumor cells with pregnenolone, 22(R)-Hydroxycholesterol [22(R)-diol] or hHDL on ROS production, phosphorylation status of p38 MAPK and cAMP response element-binding protein (CREB), CREB transcriptional activity and mRNA expression of StAR, CPY11A1/P450scc and antioxidant enzymes, superoxide dismutases [Cu,ZnSOD (SOD1), MnSOD (SOD2)], catalase (CAT) and glutathione peroxidase 1 (GPX1). We also detected the steroid product in p38 MAPK inhibitor treated Y1 cells by HPLC-MS / MS. RESULTS: Treatment of Y1 cells with H O greatly enhanced the phosphorylation of both p38 MAPK and CREB protein. Likewise, treatment of cells with pregnenolone, 22(R) diol or hHDL increased ROS production measured with the oxidation-sensitive fluorescent probe 2',7'-Dichlorofluorescin diacetate (DCFH-DA). Under identical experimental conditions, treatment of cells with these agents also increased the phosphorylation of p38 MAPK and CREB. This increased CREB phosphorylation however, was associated with its decreased transcriptional activity. The stimulatory effects of pregnenolone, 22(R)-diol and hHDL on CREB phosphorylation was abolished by a specific p38 MAPK inhibitor, SB203580. Pregnenolone, and 22(R) diol but not hHDL upregulated the mRNA expression of SOD1, SOD2 and GPX1, while down-regulated the mRNA levels of StAR and CYP11A1. The p38 inhibitor SB203580 could increase the steroid production in HDL3, 22(R)-diol or pregnenolone treated cells. CONCLUSION: Our data demonstrate induction of a ROS/p38 MAPK -mediated feedback inhibitory pathway by oxy-cholesterol and steroid intermediates and products attenuates steroidogenesis via inhibition of CREB transcriptional activity.
[Mh] Termos MeSH primário: Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo
Transdução de Sinais
Esteroides/biossíntese
Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
[Mh] Termos MeSH secundário: Neoplasias do Córtex Suprarrenal/genética
Neoplasias do Córtex Suprarrenal/metabolismo
Neoplasias do Córtex Suprarrenal/patologia
Animais
Western Blotting
Catalase/genética
Catalase/metabolismo
Linhagem Celular Tumoral
Enzima de Clivagem da Cadeia Lateral do Colesterol/genética
Enzima de Clivagem da Cadeia Lateral do Colesterol/metabolismo
Retroalimentação Fisiológica/efeitos dos fármacos
Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos
Glutationa Peroxidase/genética
Glutationa Peroxidase/metabolismo
Peróxido de Hidrogênio/farmacologia
Hidroxicolesteróis/farmacologia
Camundongos
Oxidantes/farmacologia
Fosfoproteínas/genética
Fosfoproteínas/metabolismo
Fosforilação/efeitos dos fármacos
Pregnenolona/farmacologia
Espécies Reativas de Oxigênio/metabolismo
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Superóxido Dismutase/genética
Superóxido Dismutase/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cyclic AMP Response Element-Binding Protein); 0 (Hydroxycholesterols); 0 (Oxidants); 0 (Phosphoproteins); 0 (Reactive Oxygen Species); 0 (Steroids); 0 (steroidogenic acute regulatory protein); 17711-16-9 (22-hydroxycholesterol); 73R90F7MQ8 (Pregnenolone); BBX060AN9V (Hydrogen Peroxide); EC 1.11.1.- (glutathione peroxidase GPX1); EC 1.11.1.6 (Catalase); EC 1.11.1.9 (Glutathione Peroxidase); EC 1.14.15.6 (Cholesterol Side-Chain Cleavage Enzyme); EC 1.15.1.1 (Superoxide Dismutase); EC 2.7.11.24 (p38 Mitogen-Activated Protein Kinases)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170707
[Lr] Data última revisão:
170707
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170318
[St] Status:MEDLINE
[do] DOI:10.1186/s12958-017-0239-4



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