Quantitative determination and tissue distribution of human 11β-hydroxysteroid dehydrogenase, hexose-6-phosphate dehydrogenase, glucose-6-phosphate transporter, glucocorticoid receptor and mineralocorticoid receptor mRNAs

2010 ◽  
Vol 2 (1) ◽  
Author(s):  
Shuji Ohno ◽  
Masanori Ohta ◽  
Shizuo Nakajin
Keyword(s):  
Small Intestine ◽  
Quantitative Determination ◽  
Mineralocorticoid Receptor ◽  
Hydroxysteroid Dehydrogenase ◽  
Phosphate Transporter ◽  
Physiological Effects ◽  
Wide Range

Abstract: Glucocorticoid (GC) concentrations in peripheral tissues are precisely regulated by 11β-hydroxysteroid dehydrogenase (HSD) isozymes. When considering the physiological effects of GC in various tissues, quantitative determination of these isozymes and other components involved in corticosteroid signaling is important and informative. We thus performed comprehensive determination of the expression of these mRNAs in a wide range of human tissues.: An absolute comparison of mRNA expression of human 11β-HSD isozymes, hexose-6-phosphate dehydrogenase (H6PDH), glucose-6-phosphate transporter (G6PT), glucocorticoid receptors (GRs), and mineralocorticoid receptor (MR) was performed by real-time RT-PCR.: Human 11β-HSD type 1 mRNA was strongly expressed in the liver and placenta at comparatively high levels. H6PDH was expressed at low copy number, and comparatively high expression was observed in the kidney, testis, and ovary. G6PT expression was ubiquitous, but marked expression was observed in the liver, kidney, small intestine, and colon. GRα was also ubiquitously expressed at relatively high levels, which were approximately 10-fold higher than those of MR, whereas GRβ levels were below the detection limit in all tissues. 11β-HSD type 2 was predominantly expressed in the kidney, small intestine and colon; however, copy numbers of these transcripts showed a nearly identical pattern to type 1. MR was observed in various tissues examined, but was not fully correlated to the distribution of 11β-HSD type 2.: The present quantitative results were partially consistent with previous studies. This quantification method can thus provide valuable information for understanding the physiological effects and physiological roles of glucocorticoid in humans.

Substrate and inhibitor specificity of the cloned human 11 beta-hydroxysteroid dehydrogenase type 2 isoform

1996 ◽  
Vol 270 (5) ◽  
pp. E900-E904 ◽  
Author(s):  
P. Ferrari ◽  
R. E. Smith ◽  
J. W. Funder ◽  
Z. S. Krozowski
Keyword(s):  
Dose Response ◽  
Mineralocorticoid Receptor ◽  
Response Curve ◽  
Hydroxysteroid Dehydrogenase ◽  
Significant Inhibition ◽  
Michaelis Constant ◽  
Intact Cells ◽  
Target Organs

The 11 beta-hydroxysteroid dehydrogenase type 2 (11betaHSD2) enzyme is thought to confer specificity on the mineralocorticoid receptor by inactivating glucocorticoids in mineralocorticoid target organs. The cloned 11 beta HSD2 displayed Michaelis constant values for corticosterone and cortisol of 5.1 and 61 nM, respectively. Linearity in the dose-response curve ranged between 1 and 200 nM for corticosterone and 25 and 2,000 nM for cortisol, with no evidence for complex kinetics. Inhibition of cortisol oxidation by other steroids was purely competitive in nature. Inhibition of 11 beta HSD2 activity by the end product or aldosterone occurred only at supraphysiological levels, whereas corticosterone and deoxycorticosterone displayed significant inhibition at physiological concentrations and progesterone at concentrations that occur during pregnancy. In intact transfected CHOP cells, dexamethasone was converted to 11-dehydrodexamethasone by 11 beta HSD2 but not type 1 11 beta-hydroxysteroid dehydrogenase, an aspect that may be useful in evaluating 11 beta HSD activity in intact cells.

AGN1 vs AGN2 dichotomy as seen from the point of view of ionized outflows

2019 ◽  
Vol 15 (S356) ◽  
pp. 96-96
Author(s):  
Eleonora Sani
Keyword(s):  
Accretion Rate ◽  
Point Of View ◽  
Emission Lines ◽  
Black Hole Mass ◽  
X Ray ◽  
Bolometric Luminosity ◽  
Wide Range ◽  
Strong Contrast

AbstractI present a detailed study of ionized outflows in a large sample of 650 hard X-ray detected AGN. Taking advantage of the legacy value of the BAT AGN Spectroscopic Survey (BASS, DR1), we are able to reveal the faintest wings of the [OIII] emission lines associated with outflows. The sample allows us to derive the incidence of outflows covering a wide range of AGN bolometric luminosity and test how the outflow parameters are related with various AGN power tracers, such as black hole mass, Eddington ratio, luminosity. I’ll show how ionized outflows are more frequently found in type 1.9 and type 1 AGN (50% and 40%) with respect to the low fraction in type 2 AGN (20%). Within such a framework, I’ll demonstrate how type 2 AGN outflows are almost evenly balanced between blue- and red-shifted winds. This, in strong contrast with type 1 and type 1.9 AGN outflows which are almost exclusively blue-shifted. Finally, I’ll prove how the outflow occurrence is driven by the accretion rate, whereas the dependence of outflow properties with respect to the other AGN power tracers happens to be quite mild.

scholarly journals 11?-Hydroxysteroid dehydrogenase Type 1 in obesity and Type 2 diabetes

Diabetologia ◽  
2004 ◽  
Vol 47 (1) ◽  
pp. 1-11 ◽  
Author(s):  
T. M. Stulnig ◽  
W. Waldh�usl
Keyword(s):  
Type 2 Diabetes ◽  
Hydroxysteroid Dehydrogenase

Energy Method for Determining Dynamic Characteristics of Mechanisms

1949 ◽  
Vol 16 (3) ◽  
pp. 283-288
Author(s):  
B. E. Quinn
Keyword(s):  
Dynamic Characteristics ◽  
Energy Method ◽  
Mechanical Systems ◽  
Direct Approach ◽  
Graphical Methods ◽  
Applied Forces ◽  
Complex Mechanical Systems

Abstract Two types of problems are dealt with in the paper which are involved in the design of mechanisms required to have specified dynamic characteristics: (1) Determination of applied forces required to produce specified dynamic characteristics. (2) Determination of the dynamic characteristics which will result from the application of known forces. While graphical methods may be used in the solution of type (1) problems involving more or less complex mechanical systems, they do not afford a direct approach to type (2) problems. The energy method which will be outlined can be applied in either case, although this paper will be primarily concerned with the determination of the dynamic characteristics which result when a known force is applied to a given mechanism.

scholarly journals Development and validation of the method for the quantitative determination of methotrexate in a transport medium by HPLC-MS/MS

2021 ◽  
pp. 45-51
Author(s):  
P. Yu. Mylnikov ◽  
Yu. Tranova ◽  
A. V. Shchulkin ◽  
E. N. Yakusheva
Keyword(s):  
Quantitative Determination ◽  
Gradient Elution ◽  
Sample Volume ◽  
Transport Medium ◽  
Mass Selective Detector ◽  
Transporter Protein ◽  
Wide Range ◽  
Separation Temperature

Relevance. BCRP is an efflux transporter protein that plays an important role in the pharmacokinetics of a wide range of drugs. The BCRP activity in vitro experiments is assessed by the transport of transporter protein substrates (methotrexate, etc.) across the bilipid membrane of cells overexpressingBCRP, for example, Caco-2 cells. The aim is to develop and validate a method for the quantitative determination of the BCRP substrate, methotrexate, in the transport medium of Caco-2 cells by HPLC-MS/MS. Methods. The work was performed on an Ultimate 3000 HPLC chromatograph (ThermoFisher, USA) with a TSQ Fortis tandem mass-selective detector (ThermoFisher, USA). The conditions of chromatographic analysis were as follows: column UCT Selectra C18 4.6 mm * 100 mm 5um, 100A, Selectra C18 Guard Cartridges SLC-18GDC46-5UM, separation temperature 35 °С, flow rate 0.3 ml/min, injected sample volume - 2 μl, analysis time - 10 min. Used a gradient elution: the ratio of the solution of 0.1 % formic acid and acetonitrile was at 0 min 75 and 25 %; 0.4 min 60 and 40 %; 6 minutes 20 and 80 %; 8 minutes 75 and 25 %. Under these conditions, the retention time of methotrexate is 3.11 minutes. Detection conditions: methotrexate - positive ionization mode, 455.15 m / z → 308.125 m / z, collision energy 22.99 V, source fragmentation 5, CID gas pressure 2 mTorr. The extraction of methotrexate from the transport medium (Hanks solution with 25 mM Hepes and 1% dimethyl sulfoxide) after incubation with Caco-2 cells for 3 h was carried out with a mixture of methanol + water in a ratio of 1: 1. Results. The developed method was validated according to the following parameters: selectivity, linearity, accuracy, precision, limit of quantitative determination, sample transfer, sample stability. The confirmed analytical range of the method was 60 -10,000 nmol / L in the transport medium. Conclusions: a method for the quantitative determination of methotrexate in the transport medium of Caco-2 cells by HPLC-MS / MS was developed and validated.

A Method for the Routine Determination of Corn Sirup in Prepared Meat Products

1964 ◽  
Vol 47 (5) ◽  
pp. 903-909
Author(s):  
Lester Hankin ◽  
Alphonse F Wickroski
Keyword(s):  
Statistical Analysis ◽  
Quantitative Determination ◽  
Correction Factor ◽  
Meat Products ◽  
Processed Meat ◽  
Average Amount ◽  
Wide Range ◽  
The Difference

Abstract A method has been devised for the determination of corn sirup added to processed meat products. The method is based on the quantitative determination of dextrin added to corn sirup. The dextrins are enzymatically hydrolyzed by α-amylase and β-amylase, and maltose is calculated as the difference in CuO2 found by copper reduction between a treated and an untreated aliquot. A correction factor was devised to determine the average amount of dextrin in corn sirup by testing a number of commercial sirups for their dextrin content and subjecting the data to statistical analysis. With this equation the method is applicable to a wide range of sirups. The method also permits the estimation of dextrose added to meats in excess of that included as one of the components of corn sirup.

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