Fluorescent biosensors offer a powerful tool for tracking and quantifying protein activity in living systems with high temporospatial resolution. However, the expression of genetically encoded fluorescent proteins can interfere with endogenous signaling pathways, potentially leading to developmental and physiological abnormalities. The EKAREV-NLS mouse model, which carries a FRET-based biosensor for monitoring extracellular signal-regulated kinase (ERK) activity, has been widely utilized both in vivo and in vitro across various cell types and organs. In this study, we report a significant defect in mammary epithelial development in EKAREV-NLS C57BL/6J female mice. Our findings reveal that these mice exhibit severely impaired mammary epithelial outgrowth, linked to systemic defects including disrupted estrous cycling, impaired ovarian follicle maturation, anovulation, and reduced reproductive fitness. Notably, estrogen supplementation was sufficient to enhance mammary epithelial growth in the EKAREV-NLS C57BL/6J females. Furthermore, outcrossing to the ICR genetic background fully restored normal mammary epithelial outgrowth, indicating that the observed phenotype is dependent on genetic background. We also confirmed the functional performance of the biosensor in hormone-supplemented and outcrossed tissues through time-lapse imaging of primary mammary epithelial cells. Our results underscore the critical need for thorough characterization of biosensor-carrying models before their application in specific research contexts. Additionally, this work highlights the influence of hormonal and genetic factors on mammary gland development and emphasizes the importance of careful consideration when selecting biosensor strains for mammary studies.

• Klíčová slova
• Biosensor, EKAREV–NLS mouse, ERK signaling, Estradiol supplementation, Genetic background, Hormonal imbalance, Mammary epithelium,
• MeSH
• biosenzitivní techniky * metody   MeSH
• epitelové buňky metabolismus   účinky léků   MeSH
• estrogeny * metabolismus   MeSH
• extracelulárním signálem regulované MAP kinasy metabolismus   MeSH
• genetické pozadí MeSH
• mléčné žlázy zvířat * růst a vývoj   účinky léků   MeSH
• myši inbrední C57BL * MeSH
• myši inbrední ICR MeSH
• myši transgenní * MeSH
• myši MeSH
• rezonanční přenos fluorescenční energie metody   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• estrogeny * MeSH
• extracelulárním signálem regulované MAP kinasy MeSH

Lilial (also called lysmeral) is a fragrance ingredient presented in many everyday cosmetics and household products. The concentrations of lilial in the final products is rather low. Its maximum concentration in cosmetics was limited and recently, its use in cosmetics products was prohibited in the EU due to the classification as reproductive toxicant. Additionally, according to the European Chemicals Agency, it was under assessment as one of the potential endocrine disruptors, i.e. a substance that may alter the function of the endocrine system and, as a result, cause health problems. Its ability to act as an androgen receptor agonist and the estrogenic and androgenic activity of its metabolites, to the best of our knowledge, have not yet been tested. The aim of this work was to determine the intestinal absorption, cytotoxicity, nephrotoxicity, mutagenicity, activation of cellular stress-related signal pathways and, most importantly, to test the ability to disrupt the endocrine system of lilial and its Phase I metabolites. This was tested using set of in vitro assays including resazurin assay, the CHO/HPRT mutation assay, γH2AX biomarker-based genotoxicity assay, qPCR and in vitro reporter assays based on luminescence of luciferase for estrogen, androgen, NF-κB and NRF2 signalling pathway. It was determined that neither lilial nor its metabolites have a negative effect on cell viability in the concentration range from 1 nM to 100 µM. Using human cell lines HeLa9903 and MDA-kb2, it was verified that this substance did not have agonistic activity towards estrogen or androgen receptor, respectively. Lilial metabolites, generated by incubation with the rat liver S9 fraction, did not show the ability to bind to estrogen or androgen receptors. Neither lilial nor its metabolites showed a nephrotoxic effect on human renal tubular cells (RPTEC/TERT1 line) and at the same time they were unable to activate the NF-κB and NRF2 signalling pathway at a concentration of 50 µM (HEK 293/pGL4.32 or pGL4.37). Neither lilial nor its metabolites showed mutagenic activity in the HPRT gene mutation test in CHO-K1 cells, nor were they able to cause double-strand breaks in DNA (γH2AX biomarker) in CHO-K1 and HeLa cells. In our study, no negative effects of lilial or its in vitro metabolites were observed up to 100 µM using different in vitro tests.

• MeSH
• androgeny MeSH
• biologické markery MeSH
• estrogeny toxicita   metabolismus   MeSH
• faktor 2 související s NF-E2 MeSH
• HEK293 buňky MeSH
• HeLa buňky MeSH
• hypoxanthinfosforibosyltransferasa * MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• NF-kappa B * MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• androgeny MeSH
• biologické markery MeSH
• estrogeny MeSH
• faktor 2 související s NF-E2 MeSH
• hypoxanthinfosforibosyltransferasa * MeSH
• lilial MeSH Prohlížeč
• NF-kappa B * MeSH

Catechol-O-methyl transferase (COMT) is involved in detoxification of catechol estrogens, playing cancer-protective role in cells producing or utilizing estrogen. Moreover, COMT suppressed migration potential of breast cancer (BC) cells. To delineate COMT role in metastasis of estrogen receptor (ER) dependent BC, we investigated the effect of COMT overexpression on invasion, transcriptome, proteome and interactome of MCF7 cells, a luminal A BC model, stably transduced with lentiviral vector carrying COMT gene (MCF7-COMT). 2D and 3D assays revealed that COMT overexpression associates with decreased cell invasion (p < 0.0001 for Transwell assay, p < 0.05 for spheroid formation). RNA-Seq and LC-DIA-MS/MS proteomics identified genes associated with invasion (FTO, PIR, TACSTD2, ANXA3, KRT80, S100P, PREX1, CLEC3A, LCP1) being downregulated in MCF7-COMT cells, while genes associated with less aggressive phenotype (RBPMS, ROBO2, SELENBP, EPB41L2) were upregulated both at transcript (|log2FC|> 1, adj. p < 0.05) and protein (|log2FC|> 0.58, q < 0.05) levels. Importantly, proteins driving MET signaling were less abundant in COMT overexpressing cells, and pull-down confirmed interaction between COMT and Kunitz-type protease inhibitor 2 (SPINT2), a negative regulator of MET (log2FC = 5.10, q = 1.04-7). In conclusion, COMT may act as tumor suppressor in ER dependent BC not only by detoxification of catechol estrogens but also by suppressing cell invasion and interplay with MET pathway.

• MeSH
• estrogeny metabolismus   MeSH
• katechol-O-methyltransferasa * genetika   MeSH
• katecholestrogeny MeSH
• katecholy MeSH
• nádory * MeSH
• receptory pro estrogeny metabolismus   MeSH
• tandemová hmotnostní spektrometrie MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• catechol MeSH Prohlížeč
• estrogeny MeSH
• katechol-O-methyltransferasa * MeSH
• katecholestrogeny MeSH
• katecholy MeSH
• receptory pro estrogeny MeSH

Estrogen is known to regulate bone metabolism in both women and men, but substantial gaps remain in our knowledge of estrogen and estrogen receptor alpha (ERα) regulation of adult bone metabolism. Studies using global ERα-knockout mice were confounded by high circulating sex-steroid levels, and osteocyte/osteoblast-specific ERα deletion may be confounded by ERα effects on growth versus the adult skeleton. Thus, we developed mice expressing the tamoxifen-inducible CreERT2 in osteocytes using the 8-kilobase (kb) Dmp1 promoter (Dmp1CreERT2 ). These mice were crossed with ERαfl//fl mice to create ERαΔOcy mice, permitting inducible osteocyte-specific ERα deletion in adulthood. After intermittent tamoxifen treatment of adult 4-month-old mice for 1 month, female, but not male, ERαΔOcy mice exhibited reduced spine bone volume fraction (BV/TV (-20.1%, p = 0.004) accompanied by decreased trabecular bone formation rate (-18.9%, p = 0.0496) and serum P1NP levels (-38.9%, p = 0.014). Periosteal (+65.6%, p = 0.004) and endocortical (+64.1%, p = 0.003) expansion were higher in ERαΔOcy mice compared to control (Dmp1CreERT2 ) mice at the tibial diaphysis, reflecting the known effects of estrogen to inhibit periosteal apposition and promote endocortical formation. Increases in Sost (2.1-fold, p = 0.001) messenger RNA (mRNA) levels were observed in trabecular bone at the spine in ERαΔOcy mice, consistent with previous reports that estrogen deficiency is associated with increased circulating sclerostin as well as bone SOST mRNA levels in humans. Further, the biological consequences of increased Sost expression were reflected in significant overall downregulation in panels of osteoblast and Wnt target genes in osteocyte-enriched bones from ERαΔOcy mice. These findings thus establish that osteocytic ERα is critical for estrogen action in female, but not male, adult bone metabolism. Moreover, the reduction in bone formation accompanied by increased Sost, decreased osteoblast, and decreased Wnt target gene expression in ERαΔOcy mice provides a direct link in vivo between ERα and Wnt signaling. © 2022 American Society for Bone and Mineral Research (ASBMR).

• Klíčová slova
• ESTROGENS AND SERMs, GENETIC ANIMAL MODELS, OSTEOCYTES, OSTEOPOROSIS, SEX STEROIDS,
• MeSH
• adaptorové proteiny signální transdukční metabolismus   MeSH
• alfa receptor estrogenů * genetika   metabolismus   MeSH
• dospělí MeSH
• estrogeny metabolismus   farmakologie   MeSH
• kojenec MeSH
• lidé MeSH
• messenger RNA metabolismus   MeSH
• mezibuněčné signální peptidy a proteiny metabolismus   MeSH
• myši knockoutované MeSH
• myši MeSH
• osteoblasty metabolismus   MeSH
• osteocyty * metabolismus   MeSH
• tamoxifen farmakologie   MeSH
• zvířata MeSH
• Check Tag
• dospělí MeSH
• kojenec MeSH
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• alfa receptor estrogenů * MeSH
• estrogeny MeSH
• messenger RNA MeSH
• mezibuněčné signální peptidy a proteiny MeSH
• tamoxifen MeSH

Although produced largely in the periphery, gonadal steroids play a key role in regulating the development and functions of the central nervous system and have been implicated in several chronic neuropsychiatric disorders, with schizophrenia and Alzheimer's disease (AD) most prominent. Despite major differences in pathobiology and clinical manifestations, in both conditions, estrogen transpires primarily with protective effects, buffering the onset and progression of diseases at various levels. As a result, estrogen replacement therapy (ERT) emerges as one of the most widely discussed adjuvant interventions. In this review, we revisit evidence supporting the protective role of estrogen in schizophrenia and AD and consider putative cellular and molecular mechanisms. We explore the underlying functional processes relevant to the manifestation of these devastating conditions, with a focus on synaptic transmission and plasticity mechanisms. We discuss specific effects of estrogen deficit on neurotransmitter systems such as cholinergic, dopaminergic, serotoninergic, and glutamatergic. While the evidence from both, preclinical and clinical reports, in general, are supportive of the protective effects of estrogen from cognitive decline to synaptic pathology, numerous questions remain, calling for further research.

• Klíčová slova
• Adjuvant therapy, Animal models, Cognitive deficit, Menopause, Neuroprotection, Plasticity,
• MeSH
• Alzheimerova nemoc farmakoterapie   metabolismus   MeSH
• estrogenní substituční terapie metody   MeSH
• estrogeny metabolismus   farmakologie   MeSH
• lidé MeSH
• schizofrenie farmakoterapie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• estrogeny MeSH

There is no consensus on the cutoff for positivity of estrogen receptor (ER) and progesterone receptor (PR) in endometrial cancer (EC). Therefore, we determined the cutoff value for ER and PR expression with the strongest prognostic impact on the outcome. Immunohistochemical expression of ER and PR was scored as a percentage of positive EC cell nuclei. Cutoff values were related to disease-specific survival (DSS) and disease-free survival (DFS) using sensitivity, specificity, and multivariable regression analysis. The results were validated in an independent cohort. The study cohort (n = 527) included 82% of grade 1-2 and 18% of grade 3 EC. Specificity for DSS and DFS was highest for the cutoff values of 1-30%. Sensitivity was highest for the cutoff values of 80-90%. ER and PR expression were independent markers for DSS at cutoff values of 10% and 80%. Consequently, three subgroups with distinct clinical outcomes were identified: 0-10% of ER/PR expression with, unfavorable outcome (5-year DSS = 75.9-83.3%); 20-80% of ER/PR expression with, intermediate outcome (5-year DSS = 93.0-93.9%); and 90-100% of ER/PR expression with, favorable outcome (5-year DSS = 97.8-100%). The association between ER/PR subgroups and outcomes was confirmed in the validation cohort (n = 265). We propose classification of ER and PR expression based on a high-risk (0-10%), intermediate-risk (20-80%), and low-risk (90-100%) group.

• Klíčová slova
• Cutoff, Endometrial cancer, Estrogen receptor, Progesterone receptor, Prognostic biomarker,
• MeSH
• doba přežití bez progrese choroby MeSH
• estrogeny metabolismus   MeSH
• lidé MeSH
• nádorové biomarkery metabolismus   MeSH
• nádory endometria diagnóza   metabolismus   patologie   MeSH
• přežití po terapii bez příznaků nemoci MeSH
• prognóza MeSH
• receptory pro estrogeny metabolismus   MeSH
• receptory progesteronu metabolismus   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• estrogeny MeSH
• nádorové biomarkery MeSH
• receptory pro estrogeny MeSH
• receptory progesteronu MeSH

: Worldwide, breast cancer (BC) is the most common malignancy in women, in regard to incidence and mortality. In recent years, the negative role of obesity during BC development and progression has been made abundantly clear in several studies. However, the distribution of body fat may be more important to analyze than the overall body weight. In our review of literature, we reported some key findings regarding the role of obesity in BC development, but focused more on central adiposity. Firstly, the adipose microenvironment in obese people bears many similarities with the tumor microenvironment, in respect to associated cellular composition, chronic low-grade inflammation, and high ratio of reactive oxygen species to antioxidants. Secondly, the adipose tissue functions as an endocrine organ, which in obese people produces a high level of tumor-promoting hormones, such as leptin and estrogen, and a low level of the tumor suppressor hormone, adiponectin. As follows, in BC this leads to the activation of oncogenic signaling pathways: NFκB, JAK, STAT3, AKT. Moreover, overall obesity, but especially central obesity, promotes a systemic and local low grade chronic inflammation that further stimulates the increase of tumor-promoting oxidative stress. Lastly, there is a constant exchange of information between BC cells and adipocytes, mediated especially by extracellular vesicles, and which changes the transcription profile of both cell types to an oncogenic one with the help of regulatory non-coding RNAs.

• Klíčová slova
• abdominal fat, adiponectin, breast cancer, exosomes, hormone dependency, leptin, menopause, miRNA, obesity,
• MeSH
• abdominální obezita komplikace   metabolismus   patofyziologie   MeSH
• adiponektin škodlivé účinky   metabolismus   MeSH
• estrogeny škodlivé účinky   metabolismus   MeSH
• exozom metabolismus   MeSH
• leptin imunologie   metabolismus   MeSH
• lidé MeSH
• menopauza metabolismus   MeSH
• mikro RNA metabolismus   MeSH
• nádorová transformace buněk metabolismus   MeSH
• nádorové mikroprostředí imunologie   MeSH
• nádory prsu etiologie   metabolismus   patofyziologie   MeSH
• signální transdukce genetika   MeSH
• tuková tkáň imunologie   metabolismus   MeSH
• zánět patofyziologie   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• adiponektin MeSH
• estrogeny MeSH
• exozom MeSH
• leptin MeSH
• mikro RNA MeSH

Gibbons of the genus Nomascus exhibit strong sexual dichromatism in fur color. Change of fur color in sub-adult wild Nomascus females is associated with the onset of puberty and the time of their dispersal. The variability in fur change may be influenced by social factors. In this study, we determined whether in captive females of crested gibbons begin reproductive maturity prior to dispersing and with association to their fur color. We collected 287 fecal extracts to analyze pregnandiol -3- glucuronide and 17β estradiol profiles of 4 sub-adult females (Nomascus leucogenys and Nomascus gabriellae) and 183 samples from their mothers, using enzyme immunoassays. The sub-adult females were monitored from 4 years of age. Their hormone profiles showed the onset of ovulatory cycling between 4.6 and 5.8 years. Based on the information about the estrogen influence to the secondary sex characteristic (fur color of female) the positive link between estrogen concentration and age of the sub-adult females was found. However, the amount of the estrogen can apparently be influenced by the presence of mother. If the mother was presented, the level of estrogen was higher than if the mother was missing. Our findings suggest that the probability of changing to beige fur color by the sub-adult females increased with increased age and if they were without mother. This initial study presents the maternal influence as a possible social factor affecting the fur color change of female offspring.

• MeSH
• estrogeny metabolismus   MeSH
• Hylobates fyziologie   MeSH
• Hylobatidae fyziologie   MeSH
• menstruační cyklus fyziologie   MeSH
• pigmentace * MeSH
• pohlavní dospělost MeSH
• stárnutí fyziologie   MeSH
• zvířata MeSH
• zvířecí srst fyziologie   MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• estrogeny MeSH

The equilibrium dissociation constant of competitive antagonists represents the affinity of the receptor-ligand interaction, and it is a key characteristic of many therapeutic drugs or toxic compounds. Two commonly used methods by which the affinity of the antagonist can be estimated are Schild analysis and the Cheng-Prusoff method. However, both methods yield different results when applied to systems with slopes not equal to one. The Gaddum equation, which is fundamental for both methods, should be extended to incorporate the slope parameter of the dose-response curves and this extension should diminish the differences between the Schild and Cheng-Prusoff methods. In this study, we derived a novel form of the Gaddum equation with a slope parameter (Hill coefficient) of agonist dose-response curve. We also derived the subsequent equations for Schild and Cheng-Prusoff analysis and we validated the proposed model by the measurement of several known estrogen receptor competitive antagonists. Standardized in vitro yeast reporter gene assay (BMAEREluc/ERα) has been used for the measurements and the range of used antagonist concentrations was 1.37-46.03 μM. By applying our mathematical model, both Schild and Cheng-Prusoff methods provide more similar values of antagonist affinity than the original mathematical approach. The correctness of the model has also been demonstrated by the measurement of a partial agonist with a known receptor affinity. The presented mathematical model significantly reduces the differences in values calculated by the Cheng-Prusoff and Schild methods and yields more accurate estimations of antagonist affinity.

• Klíčová slova
• Antagonists, Equilibrium constant, Gaddum equation, Inhibition curve analysis, Receptor theory, Schild analysis,
• MeSH
• alfa receptor estrogenů antagonisté a inhibitory   metabolismus   MeSH
• antagonisté estrogenového receptoru metabolismus   farmakologie   MeSH
• benzhydrylové sloučeniny metabolismus   toxicita   MeSH
• biologické modely * MeSH
• endokrinní disruptory metabolismus   toxicita   MeSH
• estradiol metabolismus   farmakologie   MeSH
• estrogeny metabolismus   farmakologie   MeSH
• fenoly metabolismus   toxicita   MeSH
• kompetitivní vazba MeSH
• lidé MeSH
• ligandy MeSH
• parciální agonismus léků MeSH
• reprodukovatelnost výsledků MeSH
• vazba proteinů MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• alfa receptor estrogenů MeSH
• antagonisté estrogenového receptoru MeSH
• benzhydrylové sloučeniny MeSH
• bisphenol A MeSH Prohlížeč
• endokrinní disruptory MeSH
• estradiol MeSH
• estrogeny MeSH
• fenoly MeSH
• ligandy MeSH

OBJECTIVES: The aim of this study was to detect endocrine disruption potential of selected bisphenols and phthalates, compare in silico prediction with results from two in vitro methods and bring up-to-date information on development of EU legislation, available in vitro methods and biomechanisms involved in endocrine disruption. MATERIAL AND METHODS: In silico approach based on the OECD QSAR Toolbox was used for prediction of estrogen receptor α binding. OECD TG 455 assay and a yeast-based YES/YAS assay was used to determine the interactions with human estrogen (ERα) and androgen receptors. RESULTS: In silico results predicted the screened phthalates as non binders and bisphenols as very strong binders of the ERα. In vitro results differed from in silico prediction in several cases but exhibited concordance mainly for strong binders of ERα. Most of the substances exhibited parallel activity (agonist-antagonist) on both estrogen and androgen receptors. Agonistic studies showed the effective concentration of 10% activity (EC10) from 5.0E-07 for strong agonists (e.g. BPC, BPTMC). Cytotoxicity was observed after 48 h exposure of S. cerevisiae to BPFL, BPG, BPM, BPTMC in concentrations starting at 3.6E-05 mol/l. CONCLUSION: Our results suggest multiple parallel interactions of tested compounds and emphasize the importance of determination of an appropriate battery of in vitro methods that will include more receptors and will be appropriate to target specific molecular mechanisms involved in endocrine disruption. Results in agonistic studies indicate agonistic potential and are supported by results of antagonistic studies with consideration of possible multiple interactions.

• MeSH
• alfa receptor estrogenů metabolismus   MeSH
• androgenní receptory metabolismus   MeSH
• androgeny metabolismus   MeSH
• biotest metody   MeSH
• endokrinní disruptory metabolismus   MeSH
• estrogeny metabolismus   MeSH
• lidé MeSH
• receptory pro estrogeny metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• alfa receptor estrogenů MeSH
• androgenní receptory MeSH
• androgeny MeSH
• endokrinní disruptory MeSH
• ESR1 protein, human MeSH Prohlížeč
• estrogeny MeSH
• receptory pro estrogeny MeSH