ENDOGENOUS NEUTRAL STEROIDS IN THE LUNG TISSUE OF EARLY AND MID-TERM HUMAN FOETUSES AND IN VITRO STUDIES ON THEIR FORMATION

1974 ◽  
Vol 75 (1) ◽  
pp. 148-158 ◽  
Author(s):  
I. Huhtaniemi
Keyword(s):  
Mass Spectrometry ◽  
Lung Tissue ◽  
Hydroxysteroid Dehydrogenase ◽  
Gas Liquid Chromatography ◽  
Wet Weight ◽  
Total Concentrations ◽  
Insight Into ◽  
Human Foetuses ◽  
Endogenous Steroids

ABSTRACT Four pools of lung tissue from 2 or 3 foetuses of 11–17 weeks' gestational age were analyzed by gas-liquid chromatography and gaschromatography-mass spectrometry for endogenous neutral steroids. All of the steroids detected were present as their mono- or disulphate conjugates. No free compounds were found. Dehydroepiandrosterone, pregnenolone and their 16α-hydroxylated derivatives were the compounds present in highest concentrations in the monosulphate fraction. The most prominent steroid in the disulphate fraction was 5-androstene-3β,17α-diol. Other compounds detected were the monosulphates of 3β,7α-dihydroxy-5-androsten-17-one, 3β,16β- dihydroxy -5- androsten-17-one, 3β,17β- dihydroxy-5-androsten-16-one, 5-androstene-3β,16β,1 7α-triol, 5-pregnene-3β,17α-diol, and 3β,17α-dihydroxy-5-pregnen-20-one and the disulphates of 5-androstene-3β,17β-diol, 3β,16α-dihydroxy-5-androsten-17-one and 5-pregnene-3β,20α-diol. The total concentrations of steroids in the lung tissue varied from 300 to 600 μg/100 g tissue wet weight. In order to gain some insight into the origin of the endogenous steroids detected, i. e. whether they were synthesized by the lung tissue or only filtered by it from the perfusing blood, minced lung tissue was incubated with dehydroepiandrosterone, pregnenolone, 3β,17α-dihydroxy-5-pregnen-20-one and with their sulphate conjugates. Evidence for the presence of the following enzymes was found: sulphokinase, 7α-hydroxylase, 16αhydroxylase, 17α-hydroxylase, 17β-hydroxysteroid dehydrogenase, 20α-hydroxysteroid dehydrogenase and C17–20 desmolase.

ENDOGENOUS STEROID SULPHATES AND GLUCURONIDES IN THE GALLBLADDER BILE FROM EARLY AND MID-TERM HUMAN FOETUSES

1973 ◽  
Vol 59 (3) ◽  
pp. 503-510 ◽  
Author(s):  
I. HUHTANIEMI
Keyword(s):  
Mass Spectrometry ◽  
Total Concentration ◽  
Gas Chromatography Mass Spectrometry ◽  
Gas Liquid Chromatography ◽  
Foetal Liver ◽  
Intestinal Contents ◽  
Wet Weight ◽  
Steroid Conjugates ◽  
Endogenous Steroid ◽  
Human Foetuses

SUMMARY The neutral steroids derived from their conjugates present in a pool of bile from 20 human foetuses were analysed by gas—liquid chromatography and gas chromatography— mass spectrometry. Ten monosulphates, twelve disulphates and seven glucuronides were detected, but no free steroids were found. Both saturated and unsaturated steroids of the C19- and C21-series were detected. The glucuronides of 5β-pregnane-3α,20α-diol, 5α-pregnane-3β,16α,20α-triol and 3α-hydroxy-5β-pregnan-20-one as well as the monosulphate of 16α-hydroxydehydroepiandrosterone were present in the highest concentrations. Other major components were the monosulphates of dehydroepiandrosterone, pregnenolone and 16α-hydroxypregnenolone and the disulphates of 5-androstene-3β,17α-diol, 3β, 17β-dihydroxy-5-androsten-16-one and 5β-prenane-3ξ,20α,21-triol. The total concentration of the monosulphate conjugates was 435 μg/100 g sample (wet weight), of the disulphates 363 μg/100 g and of the glucuronides 815 μg/100 g. Many of the compounds detected have previously been found in the foetal liver and intestinal contents. It is therefore concluded that during early and mid-gestation the foetal liver is already excreting many steroid conjugates through the biliary tract into the intestinal canal. As in other foetal compartments, considerable amounts of steroid sulphates were detected. However, the presence of steroid glucuronides in the bile suggests that the formation of hepatic glucuronides already occurs during the foetal period.

IDENTIFICATION AND MEASUREMENT OF SULPHATE-CONJUGATED NEUTRAL STEROIDS IN THE INTESTINAL CONTENTS OF EARLY AND MID-TERM FOETUSES

1973 ◽  
Vol 57 (1) ◽  
pp. 143-158 ◽  
Author(s):  
I. HUHTANIEMI ◽  
R. VIHKO
Keyword(s):  
Mass Spectrometry ◽  
Total Concentration ◽  
Fold Increase ◽  
Gas Chromatography Mass Spectrometry ◽  
Hydroxyl Groups ◽  
Gas Liquid Chromatography ◽  
Intestinal Contents ◽  
Neutral Steroid ◽  
Wet Weight ◽  
Human Foetuses

SUMMARY Sulphate-conjugated neutral steroids in the intestinal contents of early and mid-term human foetuses (14–20 weeks of gestation) were investigated by gas—liquid chromatography and gas chromatography—mass spectrometry. Twenty-four neutral steroids were found in the monosulphate fraction and 21 in the disulphate fraction. The total concentration of these steroids varied between 9·6 and 15·3 mg/100 g meconium wet weight, about half being monosulphates and half disulphates. Steroids with a 3β-hydroxy-5-ene structure were found and also saturated steroids which carried hydroxyl groups at carbons 3, 11, 15, 16, 17, 18, 19, 20 or 21. Among them were steroids not previously detected in human foetal compartments. Thus, the foetal meconium is both qualitatively and quantitatively the richest foetal source of neutral steroid sulphates so far investigated. The present results together with those reported in the literature show that in the course of pregnancy there is a many-fold increase in the steroid concentration of the foetal intestinal contents. This is most obvious in the case of saturated C19 steroids and polar pregnane derivatives. Reduction of the 20-ones to 20β-ols seems to be more extensive towards the end of pregnancy. 5, 16-Pregnadienes and 3β-hydroxy-5β-pregnanes are present in higher concentrations in early pregnancy.

METABOLISM OF ANDROGENS IN VITRO BY HUMAN FOETAL SKIN

1976 ◽  
Vol 70 (3) ◽  
pp. 491-499 ◽  
Author(s):  
F. SHARP ◽  
J. B. HAY ◽  
M. B. HODGINS
Keyword(s):  
Gestational Age ◽  
Enzyme Activities ◽  
Histochemical Study ◽  
Reductase Activity ◽  
Hydroxysteroid Dehydrogenase ◽  
Sebaceous Glands ◽  
Isomerase Activity ◽  
Human Foetuses

SUMMARY Fresh scalp, genital, chest and axillary skin from human foetuses of 12–41 weeks' maturity was incubated in Krebs' improved Ringer I medium with [7α-3H]dehydroepiandrosterone, [7α-3H]testosterone and [7α-3H]androstenedione. The metabolites identified were androstenedione, 5α-androstane-3,17-dione, androsterone, 3-epiandrosterone, 5α-dihydrotestosterone, 5α-androstane-3α,17β-diol, 5α-androstane-3β,17β-diol, 5-androstene-3β,17β-diol and testosterone. The results provide evidence for the presence of 3β-hydroxysteroid dehydrogenase, Δ4–5 isomerase, 17β-hydroxysteroid dehydrogenase, Δ4-3-oxosteroid-5α-reductase and 3α-hydroxysteroid dehydrogenase in human foetal skin. There were quantitative differences in the various enzyme activities between different body sites and skin specimens of different gestational age. 5α-Reductase activity was particularly high in genital skin. 3β-Hydroxysteroid dehydrogenase Δ4–5 isomerase activity was low in skin from a 12-week foetus, but high in skin specimens from 28-, 38- and 41-week foetuses. 17β-Hydroxysteroid dehydrogenase activity was already high in the skin of the 12-week foetus and remained so in the older foetuses. These results were correlated with the development of the foetal sebaceous glands, and were in general agreement with a parallel enzyme histochemical study. The role of androgen metabolism in human foetal skin is discussed.

STEROID FORMATION IN PORCINE OVARIAN TISSUE IN VITRO

1970 ◽  
Vol 63 (3) ◽  
pp. 441-453 ◽  
Author(s):  
Asbjørn Aakvaag
Keyword(s):  
Liquid Scintillation ◽  
Specific Activity ◽  
Ovarian Tissue ◽  
Gas Liquid Chromatography ◽  
Similar Concentration ◽  
Endogenous Substrates ◽  
Exogenous Progesterone ◽  
Relative Utilization ◽  
Radioactive Substrate

ABSTRACT Slices of non-luteinized porcine ovaries have been incubated in the presence or absence of human chorionic gonadotrophin (HCG) and exogenous radioactive substrates. Progesterone, 17α-hydroxyprogesterone and androstenedione were isolated in a radiochemically pure form. The chemical mass and the specific activity were determined by gas liquid chromatography and liquid scintillation spectrometry. HCG stimulated the rate of formation of androstenedione in the absence of exogenous substrates with a factor of 4–8. In the presence of pregnenolone or progesterone at a concentration of about 2 × 10−6 mol/l the stimulatory effect of HCG was either abolished or markedly reduced. The conversion of exogenous progesterone to androstenedione was reduced in response to HCG indicating that the capacity of the tissue to convert progesterone to androstenedione was limited, and that the limit was reached at this rather low substrate concentration. These findings furthermore suggest that the endogenous rather than the exogenous radioactive substrate will be »preferred« by the tissue. The observations demonstrate the necessity of measuring both the radioactivity and the chemical mass of the products in investigations of this type using radioactive substrates. The formation of progesterone from endogenous substrates was also stimulated by HCG. [1-14C] acetate and [7α-3H]cholesterol were not utilized by the tissue for steroid formation. Exogenous [4-14C] pregnenolone and [7α-3H] progesterone in similar concentration were both utilized for production of 17α-hydroxyprogesterone and androstenedione. HCG had no effect on the relative utilization of the radioactive substrates.

Oxygenation of 18-hydroxycorticosterone as the final reaction for aldosterone biosynthesis

1984 ◽  
Vol 107 (3) ◽  
pp. 395-400 ◽  
Author(s):  
Itaru Kojima ◽  
Etsuro Ogata ◽  
Hiroshi Inano ◽  
Bun-ichi Tamaoki
Keyword(s):  
Carbon Monoxide ◽  
Hydroxysteroid Dehydrogenase ◽  
Dependent Manner ◽  
In Vitro Production ◽  
Mitochondrial Preparation ◽  
Final Reaction ◽  
Vitro Production ◽  
Dose Dependent ◽  
Cytochrome P 450

Abstract. Incubation of 18-hydroxycorticosterone with the sonicated mitochondrial preparation of bovine adrenal glomerulosa tissue leads to the production of aldosterone, as measured by radioimmunoassay. The in vitro production of aldosterone from 18-hydroxycorticosterone requires both molecular oxygen and NADPH, and is inhibited by carbon monoxide. Cytochrome P-450 inhibitors such as metyrapone, SU 8000. SU 10603, SKF 525A, amphenone B and spironolactone decrease the biosynthesis of aldosterone from 18-hydroxycorticosterone. These results support the conclusion that the final reaction in aldosterone synthesis from 18-hydroxycorticosterone is catalyzed by an oxygenase, but not by 18-hydroxysteroid dehydrogenase. By the same preparation, the production of [3H]aldosterone but not [3H]18-hydroxycorticosterone from [1,2-3H ]corticosterone is decreased in a dose-dependent manner by addition of non-radioactive 18-hydroxycorticosterone.

Pertumbuhan Dan Perkembangan Anggrek Dendrobium anosmum Pada Media Kultur In Vitro Dengan Beberapa Konsentrasi Air Kelapa

Agrologia ◽  
2018 ◽  
Vol 1 (1) ◽  
Author(s):  
S. Tuhuteru ◽  
Meity L Hehanussa ◽  
Simon H.T Raharjo
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Water Concentration ◽  
Medium Composition ◽  
Coconut Water ◽  
Orchid Species ◽  
Randomized Design ◽  
Wet Weight ◽  
Completely Randomized Design

Dendrobium anosmum is one of natural orchids in Indonesia. Optimization of medium composition for orchid propagation through in vitro culture is necessary to enhance propagule multiplication capabilities and quality. This study was aimed to study the influence of concentration of coconut water in culture medium on in vitro growth and development of D. anosmum orchid species and to determine the optimal coconut water concentration in culture media.  The experiment were arranged in a Completely Randomized Design with four treatments and eight replications. The treatments consisted of the addition of coconut water with concentrations: 0 ml•l -1 (control), 50 ml•l-1, 100 ml•l-1 and 150 ml•l-1. The results showed that addition of coconut water in culture medium gave different effect on shoot growth and multiplication of D. anosmum orchids.  Coconut water concentration of 100 ml•l-1 was the best concentration for growth and multiplication of D. anosmum orchids, based on both shoots and roots growth, plantlet height and wet weight.

Obtaining and Characterization of Alhagi persarum Boiss. et Buhse Callus Cell Cultures, Isoflavonoid Producers

2020 ◽  
Vol 36 (6) ◽  
pp. 35-48
Author(s):  
D.V. Коchkin ◽  
G.I. Sobolkovа ◽  
А.А. Fоmеnkov ◽  
R.А. Sidorov ◽  
А.М. Nоsоv
Keyword(s):  
Fatty Acids ◽  
Cell Culture ◽  
Liquid Chromatography ◽  
Secondary Metabolites ◽  
Cell Cultures ◽  
Mass Spectrometric ◽  
Mass Spectrometric Detection ◽  
Gas Liquid Chromatography ◽  
Callus Cell

The physiological characteristics of the callus cell cultures of Alhagi persarum Boiss et Buhse, a member of the legume family, widely used in folk medicine, have been studied. It was shown that the source of the explant was an important factor in the initiation of callusogenesis: more intense callusogenesis (almost 100%) was observed for explants from various organs of sterile seedlings, rather than intact plants (less than 30%). As a result, more than 20 lines of morphologically different callus cell cultures were obtained, and the growth parameters for the 5 most intensively growing lines were determined. The composition of fatty acids (FA) of total lipids and secondary metabolites in the most physiologically stable callus line Aр-207 was analyzed. Using capillary gas-liquid chromatography with mass spectrometric detection (GLC-MS), 19 individual C12--C24 FAs were identified, the main fraction of which were palmitic (~ 23%), stearic (~ 22%), linoleic (~ 14%) and α-linolenic (~ 33%) acids. The established atypical ratio of FAs (a simultaneous high content of both saturated FAs and polyunsaturated α-linolenic acid) is possibly due to the adaptation of cells to in vitro growth conditions. Phytochemical analysis of the secondary metabolites was carried out using ultra-performance liquid chromatography with electrospray ionization mass spectrometric detection (UPLC MS). Compounds belonging to different structural groups of isoflavones were found. Aglycones (calycosin, formononetin and afrormosin isomer), glucosides (formononetin glucoside), as well as esters of glucosides (malonylglycosides of calicosin, formononetin, afrormosin isomers, glycitein and genistein) were detected. These secondary metabolites are widespread in plants of the Fabaceae family; however, isoflavones are rare in representatives of the Alhagi genus. The presence of malonylated isoflavone glycosides in Alhagi spp. was shown for the first time. endemic plant species, Alhagi, in vitro cell culture, callus cell culture, isoflavones, fatty acids All studies were carried out using the equipment of the "Experimental Biotechnological Facility" and the "All-Russian Collection of Cell Cultures of Higher Plants" of IРР RAS. This work was supported by the Russian Foundation for Basic Research (RFBR), contract no.18-54-06021 (Az_a), and the Government of the Russian Federation, Megagrant Project no. 075-15-2019-1882.

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