Assessment of agonistic and antagonistic properties of humidifier disinfectants to the estrogenic and androgenic receptors by transactivation assay

2021 ◽  
Author(s):  
Handule Lee ◽  
Juyoung Park ◽  
Kwangsik Park
Keyword(s):  
Transactivation Assay

scholarly journals Nucleocytoplasmic Shuttling of Endocytic Proteins

2001 ◽  
Vol 153 (7) ◽  
pp. 1511-1518 ◽  
Author(s):  
Manuela Vecchi ◽  
Simona Polo ◽  
Viviane Poupon ◽  
Jan-Willem van de Loo ◽  
Alexandre Benmerah ◽  
...  
Keyword(s):  
Transcriptional Regulation ◽  
Nuclear Export ◽  
Myeloid Leukemia ◽  
Nuclear Translocation ◽  
Initial Rate ◽  
Nucleocytoplasmic Shuttling ◽  
Cellular Processes ◽  
Transactivation Assay ◽  
Ordered Assembly ◽  
Endocytic Proteins

Many cellular processes rely on the ordered assembly of macromolecular structures. Here, we uncover an unexpected link between two such processes, endocytosis and transcription. Many endocytic proteins, including eps15, epsin1, the clathrin assembly lymphoid myeloid leukemia (CALM), and α-adaptin, accumulate in the nucleus when nuclear export is inhibited. Endocytosis and nucleocytoplasmic shuttling of endocytic proteins are apparently independent processes, since inhibition of endocytosis did not appreciably alter nuclear translocation of endocytic proteins, and blockade of nuclear export did not change the initial rate of endocytosis. In the nucleus, eps15 and CALM acted as positive modulators of transcription in a GAL4-based transactivation assay, thus raising the intriguing possibility that some endocytic proteins play a direct or indirect role in transcriptional regulation.

scholarly journals The Alginate Immobilization of Metabolic Enzymes Platform Retrofits an Estrogen Receptor Transactivation Assay With Metabolic Competence

2020 ◽  
Vol 178 (2) ◽  
pp. 281-301
Author(s):  
Chad Deisenroth ◽  
Danica E DeGroot ◽  
Todd Zurlinden ◽  
Andrew Eicher ◽  
James McCord ◽  
...  
Keyword(s):  
Estrogen Receptor ◽  
Biological Effects ◽  
Parent Compound ◽  
False Negative ◽  
Metabolic Enzymes ◽  
Vitro Assay ◽  
Transactivation Assay ◽  
Alginate Immobilization ◽  
Active Substances ◽  
Receptor Transactivation

Abstract The U.S. EPA Endocrine Disruptor Screening Program utilizes data across the ToxCast/Tox21 high-throughput screening (HTS) programs to evaluate the biological effects of potential endocrine active substances. A potential limitation to the use of in vitro assay data in regulatory decision-making is the lack of coverage for xenobiotic metabolic processes. Both hepatic- and peripheral-tissue metabolism can yield metabolites that exhibit greater activity than the parent compound (bioactivation) or are inactive (bioinactivation) for a given biological target. Interpretation of biological effect data for both putative endocrine active substances, as well as other chemicals, screened in HTS assays may benefit from the addition of xenobiotic metabolic capabilities to decrease the uncertainty in predicting potential hazards to human health. The objective of this study was to develop an approach to retrofit existing HTS assays with hepatic metabolism. The Alginate Immobilization of Metabolic Enzymes (AIME) platform encapsulates hepatic S9 fractions in alginate microspheres attached to 96-well peg lids. Functional characterization across a panel of reference substrates for phase I cytochrome P450 enzymes revealed substrate depletion with expected metabolite accumulation. Performance of the AIME method in the VM7Luc estrogen receptor transactivation assay was evaluated across 15 reference chemicals and 48 test chemicals that yield metabolites previously identified as estrogen receptor active or inactive. The results demonstrate the utility of applying the AIME method for identification of false-positive and false-negative target assay effects, reprioritization of hazard based on metabolism-dependent bioactivity, and enhanced in vivo concordance with the rodent uterotrophic bioassay. Integration of the AIME metabolism method may prove useful for future biochemical and cell-based HTS applications.

Bioactivation of Quinolines in a Recombinant Estrogen Receptor Transactivation Assay Is Catalyzed by N-Methyltransferases

2019 ◽  
Vol 32 (4) ◽  
pp. 698-707
Author(s):  
Markus Brinkmann ◽  
Bogdan Barz ◽  
Danielle Carrière ◽  
Mirna Velki ◽  
Kilian Smith ◽  
...  
Keyword(s):  
Estrogen Receptor ◽  
Transactivation Assay ◽  
Receptor Transactivation

scholarly journals Transactivation Assay for Determination of Glucocorticoid Bioactivity in Human Serum

2002 ◽  
Vol 87 (8) ◽  
pp. 3740-3744 ◽  
Author(s):  
Taneli Raivio ◽  
Jorma J. Palvimo ◽  
Senja Kannisto ◽  
Raimo Voutilainen ◽  
Olli A. Jänne
Keyword(s):  
Human Serum ◽  
Transactivation Assay

515: P53 protein evolutionary functional divergence through the lens of a yeast-based transactivation assay

2014 ◽  
Vol 50 ◽  
pp. S124
Author(s):  
I. Raimondi ◽  
M. Lion ◽  
S. Donati ◽  
O. Jousson ◽  
Y. Ciribilli ◽  
...  
Keyword(s):  
P53 Protein ◽  
Functional Divergence ◽  
Transactivation Assay

5-Aminosalicylic Acid, A Weak Agonist for Aryl Hydrocarbon Receptor That Induces Splenic Regulatory T Cells

Pharmacology ◽  
2021 ◽  
pp. 1-7
Author(s):  
Atsuhito Kubota ◽  
Masaru Terasaki ◽  
Rie Takai ◽  
Masaki Kobayashi ◽  
Ryuta Muromoto ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Aryl Hydrocarbon Receptor ◽  
Flow Cytometry Analysis ◽  
Aminosalicylic Acid ◽  
Aryl Hydrocarbon ◽  
Transactivation Assay ◽  
Splenic Cells ◽  
Inflammatory Bowel

<b><i>Introduction:</i></b> 5-Aminosalicylic acid (5-ASA) is widely used as a key drug in inflammatory bowel disease. It has been recently reported that 5-ASA induces CD4 + Foxp3 + regulatory T cells (Tregs) in the colon via the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor that regulates inflammation. However, the role of 5-ASA as an AhR agonist that induces Tregs in the spleen remains unknown. <b><i>Methods:</i></b> In the present study, we investigated these themes using an AhR-mediated transactivation assay and flow cytometry analysis. The experiments were conducted by using DR-EcoScreen cells and C57BL/6 mice. <b><i>Results:</i></b> The DR-EcoScreen cell-based transactivation assay revealed that 5-ASA acted as a weak AhR agonist at concentrations of ≥300 μM (1.31–1.45-fold), and that a typical AhR agonist, 2,3,7,8-tetrachlorodibenzo-<i>p</i>-dioxin (TCDD), activated AhR at a concentration of 0.1 nM (22.8-fold). In addition, the treatment of mouse splenic cells with 300 μM 5-ASA in a primary culture assay significantly induced CD4+CD25 + Foxp3 + Tregs (control vs. 5-ASA: 9.0% vs. 12.65%, <i>p</i> &#x3c; 0.05), while 0.1 nM TCDD also showed significant induction of Tregs (control vs. TCDD: 9.0% vs. 14.1%, <i>p</i> &#x3c; 0.05). Interestingly, this induction was eliminated by co-treatment with an AhR antagonist, CH-223191. <b><i>Discussion:</i></b> These results suggest that 5-ASA is a weak agonist of AhR and thereby induces Tregs in spleen cells. Our findings may provide useful insights into the mechanism by which 5-ASA regulates inflammation.

scholarly journals Use of Cryopreserved Transiently Transfected Cells in High-Throughput Pregnane X Receptor Transactivation Assay

2007 ◽  
Vol 12 (2) ◽  
pp. 248-254 ◽  
Author(s):  
Zhengrong Zhu ◽  
Jaime Puglisi ◽  
David Connors ◽  
Jeremy Stewart ◽  
John Herbst ◽  
...  
Keyword(s):  
High Throughput ◽  
Hepg2 Cells ◽  
Pregnane X Receptor ◽  
Cell Permeability ◽  
Excellent Correlation ◽  
Transfected Cells ◽  
Transactivation Assay ◽  
Assay Performance ◽  
Cytotoxic Compounds ◽  
Receptor Transactivation

Cryopreserved, transiently transfected HepG2 cells were compared to freshly transfected HepG2 cells for use in a pregnane X receptor (PXR) transactivation assay. Assay performance was similar for both cell preparations; however, cryopreserved cells demonstrated less interassay variation. Validation with drugs of different PXR activation potencies and efficacies demonstrated an excellent correlation ( r2 > 0.95) between cryopreserved and fresh cells. Cryopreservation did not change the effect of known CYP3A4 inducers that have poor cell permeability, indicating that cryopreservation had little effect on membrane permeability. In addition, cryopreserved HepG2 cells did not exhibit enhanced susceptibility to cytotoxic compounds compared to transiently transfected control cells. The use of cryopreserved cells enables this assay to run with enhanced efficiency.

scholarly journals Tat-Neutralizing Antibodies in Vaccinated Macaques

2003 ◽  
Vol 77 (5) ◽  
pp. 3157-3166 ◽  
Author(s):  
Ilia Tikhonov ◽  
Tracy J. Ruckwardt ◽  
Glen S. Hatfield ◽  
C. David Pauza
Keyword(s):  
Neutralizing Antibodies ◽  
Vaccine Antigen ◽  
Amino Terminus ◽  
Tat Protein ◽  
Strong Response ◽  
Basic Domain ◽  
Transactivation Assay ◽  
Clade B ◽  
Clade C ◽  
A Cell

ABSTRACT The human immunodeficiency virus Tat protein is essential for virus replication and is a candidate vaccine antigen. Macaques immunized with Tat or chemically modified Tat toxoid having the same clade B sequence developed strong antibody responses. We compared these antisera for their abilities to recognize diverse Tat sequences. An overlapping peptide array covering three clade B and two clade C Tat sequences was constructed to help identify reactive linear epitopes. Sera from Tat-immunized macaques were broadly cross-reactive with clade B and clade C sequences but recognized a clade B-specific epitope in the basic domain. Sera from Tat toxoid-immunized macaques had a more restricted pattern of recognition, reacting mainly with clade B and with only one clade B basic domain sequence, which included the rare amino acids RPPQ at positions 57 to 60. Monoclonal antibodies against the amino terminus or the domain RPPQ sequence blocked Tat uptake into T cells and neutralized Tat in a cell-based transactivation assay. Macaques immunized with Tat or Tat toxoid proteins varied in their responses to minor epitopes, but all developed a strong response to the amino terminus, and antisera were capable of neutralizing Tat in a transactivation assay.

scholarly journals A New Vector Set for GAL4-Dependent Transactivation Assay in Plants.

1998 ◽  
Vol 15 (4) ◽  
pp. 217-220 ◽  
Author(s):  
Yoshiharu Y. Yamamoto ◽  
Xing-Wang Deng
Keyword(s):  
Transactivation Assay
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