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  • Articles  (16)
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  • Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry  (16)
  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 3 (1986), S. 109-126 
    ISSN: 0739-4462
    Keywords: Ecdysteroid phosphoester ; Manduca sexta ; midgut ; C18 SEP-PAK ; β-glucuronidase ; sulphatase ; acid phosphatase ; ATP:ecdysteroid phosphotransferase ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In incubations with 80,000g supernatant of Manduca sexta midgut homogenates, [3H]ecdysone was converted to 3-[3H]epiecdysone and tritiumlabeled highly polar metabolites. C18 SEP-PAK cartridges were found suitable for the separation and purification of the free ecdysteroids and of the highly polar metabolites. Eighty to ninety percent of the metabolites were hydrolyzed by enzyme mixtures (mainly β-glucuronidase, sulphatase, and acid phosphatase) from molluscs, even when β-glucuronidase activity was completely inhibited by D-saccharic acid 1,4-lactone, or various human acid phosphatases (free of sulphatase activity). In each experiment, the hydrolysate contained a much higher proportion of 3-epiecydsone than the free (unconjugated) ecdysteroid fraction. [3H]ecdysone was not metabolized in anaerobic incubations of midgut supernatant that had been filtered through Sephadex G-25. Addition of 5 mM ATP and 5 mM Mg2+ restored the conjugate formation in incubations of Sephadex-filtered supernatant. Four ecdysone conjugates and two 3-epiecdysone conjugates were resolved by reversedphase ion-pair high-performance liquid chromatography. It is concluded that the midgut cytosol contains several ATP:ecdysteriod phosphotransferases. This is the first demonstration of the formation of ecdysteroid phosphoconjugates in a cell-free system.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 3 (1986), S. 415-421 
    ISSN: 0739-4462
    Keywords: campesterol ; cholesterol ; sitosterol ; 24-methylenecholesterol ; ecdysteroids ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In an effort to determine the sterol precursor(s) of the 28-carbon ecdysteroid, makisterone A, honey bee pupae (13 days post-oviposition) were injected with radiolabeled sterols and subsequently examined for labeled ecdysteroids. High performance liquid chromatography of the pupal extracts revealed that [3H]campesterol was converted to a compound that behaved chromatographically identical to authentic makisterone A, and [14C]cholesterol was incorporated into a compound chromatographically like 20-hydroxyecdysone. No incorporation of either 24-[3H]methylenecholesterol or [14C]sitosterol into an ecdysteroid was observed. The neutral sterols of uninjected honey bee pupae contained 49.8% 24-methylenecholesterol on a relative percent basis and, with three other C28 and C29 sterols, accounted for over 99% of the total sterols present.
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  • 3
    ISSN: 0739-4462
    Keywords: tobacco hornworm ; epiecdysteroids ; ecdysteroid conjugates ; 3-epi-20-hydroxyecdysonoic acid ; mass spectrometry ; NMR spectroscopy ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: [14C]Cholesterol was injected into fifth-instar larvae of Manduca sexta, and the metabolites were isolated and identified from 8-day-old male and female pupae. A major portion of the metabolized cholesterol was esterified either with a sulfate group or with fatty acids. The predominant ecdysteroid metabolites were 20-hydroxyecdysone, 20,26-dihydroxyecdysone, 20-hydroxyecdysonoic acid, and 3-epi-20-hydroxyecdysonoic acid. Smaller amounts of ecdysteroids were identified as conjugates of 26-hydroxyecdysone, 3-epi-20-hydroxyecdysone, 20,26-dihydroxyecdysone, and its 3α-epimer. The metabolic profiles were similar for both male and female pupae. The two ecdysteroid acids were identified by nuclear magnetic resonance spectroscopy and chemical ionization mass spectrometry and by mass spectral analyses of their methyl esters. Detection of 3-epi-20-hydroxyecdysonoic acid as a major metabolite is significant, as its occurrence has been scarcely reported. 3-Epiecdysteroid acid formation is discussed as a possible ecdysteroid-inactivating pathway that may be operating specifically in lepidopterous insects or in particular developmental stages such as eggs or pupae.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 12 (1989), S. 201-218 
    ISSN: 0739-4462
    Keywords: 3-epimerization ; 3-dehydroecdysone ; 3-epiecdysone ; 3α-hydroxyecdysteroids ; 3β-hydroxyecdysteroids ; NADH ; NADPH ; molting hormone inactivation ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Ecdysone and 20-hydroxyecdysone are converted to their 3-epimers by enzymes in the midgut cytosol of Manduca sexta larvae. A partially purified cytosol preparation has been used to analyze the nature of and the interaction between these enzymes. The cytosol was shown to contain ecdysone oxidase, one or more 3-oxoecdysteroid 3α-reductase(s), and one or more 3-oxoecdysteroid 3β-reductase(s). The reductases reacted at different velocities with NADH and NADPH. With NADH, 3α-reduction was the major reaction; with NADPH, 3β-reduction was the major reaction. The apparent kinetic parameters for the enzymes support the assumed two-step mechanism for the 3-epimerization with a 3-oxoecdysteroid as intermediate.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 1 (1984), S. 323-330 
    ISSN: 0739-4462
    Keywords: gypsy moth ; pupal ecdysteroids ; high-pressure liquid chromatography ; radioimmunoassay ; mass spectrometry ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Normal and reverse-phase high-performance liquid chromatography in conjunction with radioimmunoassay and mass spectrometry were used to identify the free and conjugated ecdysteroids (after enzymatic hydrolysis) from day-4 pupae of the gypsy moth, Lymantria dispar L. Seven ecdysteroids were searched for, but only 20-hydroxyecdysone (964 ng/g fresh weight) and ecdysone (367 ng/g fresh weight) were detected. Analysis of conjugated ecdysteriods after liberation by hydrolysis also indicated the presence of 20-hydroxyecdysone (21.6 ng/g fresh weight) and ecdysone (2.4 ng/g fresh weight). Neither 26-hydroxyecdysone nor the 3α-epimers of 20-hydroxyecdysone or ecdysone were detected.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 3 (1986), S. 235-252 
    ISSN: 0739-4462
    Keywords: ticks ; Ornithodoros moubata ; metabolism of ingested ecdysteroids ; detoxification ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Ornithodoros moubata females proved to be extremely sensitive to ingested 22,25-dideoxyecdysone; 15-20 ng provoked molting in all females and temporarily inhibited vitellogenesis. In contrast, this tick was very resistant to ingested ecdysteroids containing 22-OH groups, such as ecdysone, 20-hydroxyecdysone, ponasterone A, and makisterone A. Dosages about 500 times greater were necessary to produce supermolting and reduce fecundity [Connat et al: Z Ang Ent 96, 520 (1983)]. Ingested tritiated ecdysone, 20-hydroxyecdysone, 2-deoxyecdysone, and ponasterone A were rapidly converted to apolar esterase-labile metabolites having approximately the same retention time as the AP2 identified as esters of ecdysteroids at C-22 with long-chain fatty acids (C16:0, C18:0, C18:1, C18:2) [Diehl et al: Int J Invert Reprod Dev 8, 1 (1985)]. These products were then gradually transformed to the more polar apolar conjugates, AP1. A more detailed study with ingestion of large quantities of 20-hydroxyecdysone (10 μ/ml blood) demonstrated that only small amounts of free hormone were present in the hemolymph during the first day after the blood meal. The hormone was rapidly metabolized to AP2, then to AP1, in the intestinal cells and to a lesser extent in the peripheral tissues. Finally, AP1 accumulated in the intestinal cells and midgut content, probably because excretion outside the animal is impossible in this tick species.In contrast, ingested 22,25-dideoxyecdysone was not metabolized to apolar products. This could account for its high biological activity. This compound was converted to unidentified more polar products. Two of them comigrated with ecdysone and 20-hydroxyecdysone on RP-18 HPLC column, but not on silica column, and therefore cannot correspond to these compounds.We hypothesize that esterification of ecdysteroids at the C-22 position with fatty acids represents a detoxification mechanism for ingested ecdysteroids that might be present in blood from herbivorous or parasite-infected hosts.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 13 (1990), S. 127-143 
    ISSN: 0739-4462
    Keywords: Cotesia congregata ; NMR ; 13C ; fat body metabolism ; pyruvate ; Krebs cycle ; lipogenesis ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Metabolism of fifth instar Manduca sexta larvae was significantly altered during parasitization by Cotesia congregata. The in vivo 13C natural abndance spectrum and histological examination of the fat body demonstrated significantly reduced fat levels and increased deposition of glycogen in parasitized insects. 1H NMR analysis demonstrated that the free amino acid level was reduced in the hemolymph of parasitized larvae, but control and parasitized insects had similar levels of trehalose. [13C]2-pyruvate injected into the dorsal vessel was incorporated into numerous metabolites including amino acids, Krebs cycle intermediates, trehalose, and fat. In control larvae pyruvate was incorporated primarily into fat following oxidation to CoASAc. Pyruvate was metabolized slower in parasitized larvae and following decarboxylation to oxaloacetate was metabolized by Krebs cycle and incorporated into amino acids. Only small amounts of trehalose were synthesized from pyruvate in control and parasitized larvae and glycogen synthesis was not observed. Analysis of hemolymph indicated no net synthesis of glucose by gluconeogenesis. Control and parasitized larvae incorporated [13C]1-glucose primarily into trehalose. Control larvae also synthesized fat from gulcose. In contrast, fat synthesis was not observed in parasitized insects which incorporated glucose into glycogen. Differences in glucose metabolism, therefore, appeared to explain the difference in storage metabolite levels between control and parasitized larvae.
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  • 8
    ISSN: 0739-4462
    Keywords: 26-hydroxyecdysone 26-phosphate ; enzymatic hydrolysis ; ecdysteroid phosphate isolation ; ecdysteroid conjugate ; Manduca sexta ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The major ecdysteroid conjugate present in eggs (48-64 h old) of the tobacco hornworm has been purified by XAD-2 chromatography, C18 SEP-PAK separations, and ion suppression reversed-phase high-performance liquid chromatography. Enzymatic hydrolysis of the conjugate with acid phosphatase from human seminal fluid gave 26-hydroxyecdysone. The conjugate was identified as 26-hydroxyecdysone 26-phosphate by nuclear magnetic resonance and fast atom bombardment mass spectrometry. This compound is also the major conjugate of newly laid eggs (0-1 h old) of the tobacco hornworm. The role for ecdysteroid conjugates is discussed.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 4 (1987), S. 57-66 
    ISSN: 0739-4462
    Keywords: 25-azasteroids ; insect development ; desmosterol ; sitosterol dealkylation ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Larvae of Spodoptera litura (F.) grown on an artificial diet completed larval development in 19.2 days and attained a maximum weight of 873.2 mg. When fed dietary concentrations of 50 ppm of 25-azacholesterol or 10 ppm of 25-azacholestane, the larval developmental period increased to 28.4 and 23.4 days, and the larval weights were 447.5 and 542.3 mg, respectively. Both compounds induced distinct melanization effects and caused production of larval-pupal intermediates and severe mortality. Treatments with concentrations of 50 ppm or more of either azasteroid caused a decline in pupal period and earlier eclosion and emergence of abnormal adults. Egg laying and hatchability decreased with increasing concentrations of azasteroids in the larval diets.When 1 ppm or more of 25-azasteroid is added to the artificial diet, the insect larvae contain identifiable amounts of desmosterol, in addition to cholesterol, campesterol, and sitosterol, which are present in Spodoptera grown on artificial diet alone. Desmosterol accumulation in the insect body is due to an inhibition of the Δ24-sterol reductase by 25-azasteroids. An increase in the concentration of these azasteroids in the diet results in an increase in sitosterol concentration and simultaneous reduction in the cholesterol levels due to inhibition of conversion of sitosterol. This inhibition appears to be more pronounced with 25-azacholestane treatment than with 25-azacholesterol.
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  • 10
    ISSN: 0739-4462
    Keywords: radiolabeled ecdysteroids ; conjugates ; 26-hydroxyecdysone 22-glucoside ; 26-hydroxyecdysone 26-phosphate ; 26-hydroxyecdysone ; 20-26-dihydroxyecdysone ; 3-epi-20-26-dihydroxyecdysone ; 3-epi-20-hydroxyecdysonoic-acid ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The levels of both free and conjugated ecdysteroids, maternally labeled from [14C]cholesterol, of six different age groups of Manduca sexta eggs were quantitatively determined. Eggs 0-1-h old contain about 2.5 and 35 μ/g of the 2- and 26-phosphates of 26-hydroxyecdysone, respectively, and 1 μg/g of 26-hydroxyecdysone. During embryogenesis of 26-hydroxyedcdysone 26-phosphate is hydrolyzed to 26-hydroxyecdysone, which reaches a peak titer in 1-18-h-old eggs; the level of 26-hydroxyecdysone 2-phosphate remains rather constant. Additionally, other metabolic modifications such as hydroxylation, conjugation, epimerization, and oxidation are occurring; and as the level of the 26-hydroxyecdysone 26-phosphate decreases there is a progression of other ecdysteroids. C-20 hydroxylation first appears in 24-40-h-old eggs and reaches peak activity in 48-64-h-old eggs, where 20-hydroxyecdysone and 20, 26-dihydroxyecdysone are both present at peak titer but the latter is the major free ecdysteroid. Ecdysone is observed at measurable levels only in the three age groups of eggs between 1 and 64 h-old. C-3 epimerase activity also appears at 24-40 h and continually increases throughout embryogenesis to the point that 3-epi-26-hydroxyecdysone and 3-epi-20, 26-dihydroxyecdysone are the major free ecdysteroids in 96-h-old eggs. A new ecdysteroid conjugate, 26-hydroxyecdysone 22-glucoside, first appears at 24-40h and becomes the major conjugate in 72-80-h-old eggs; it represents an apparent end-product as its peak titer is reached and maintained throughout the final embryonic stages. 20-Hydroxyecdysonoic acid occurs in 48-64-h-old eggs, and along with 3-epi-20-hydroxyecdysonoic and ecdysonoic acids in 72-88-h-old eggs. 20-Hydroxyecdysonoic acid peaks during the latter time interval, and as its titer subsequently falls, there is a concurrent increase in the level of 3-epi-20-hydroxyecdysonoic which was identified as the second major component of the ecdysteroid conjugate fraction of 0-1-h-old larvae. Our results indicate that there is little or no biosynthesis of ecdysteroids during embryogenesis; that the materal ecdysteroid conjugate 26-hydroxyecdysone 26-phosphate serves as source for 26-hydroxyecdysone and the numerous metabolites; that 26-hydroxyecdysone and 20,26-dihydroxyecdysone may be the active hormones during embryonic development; and that glucosylation, epimerization, and formation of acids cosntitute inactivation processes. A scheme of the proposed pathways involved in the metabolism of 26-hydroxyecdysone 26-phosphate in the developing eggs of m. sexta is presented.
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