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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 12 (1989), S. 63-74 
    ISSN: 0739-4462
    Keywords: [14C]cholesterol ; HPLC ; ecdysteroid titers ; ecdysteroid metabolism ; epimerization ; hydroxylation ; oxidation ; epiecdysteroid acids ; epiecdysteroid phosphates ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The levels of individual free and conjugated ecdysteroids and ecdysteroid acids, labeled from [14C]cholesterol, in five different age groups of male Manduca sexta during pupal-adult development were determined by HPLC. Eight free ecdysteroids, eight ecdysteroid phosphates, and two ecdysteroid acids were identified. Newly ecdysed pupae contained predominantly 3-epiecdysteroids in each of the free, conjugated, and acidic ecdysteroid fractions. The titer of each ecdysteroid fraction rose sharply by day 4, and this was particularly noteworthy with respect to free ecdysone and 3-epi-20-hydroxyecdysonoic acid. This stage demonstrated high degrees of ecdysone biosynthesis, oxidative catabolism, and phosphorylation. As development proceeded to day 16, total ecdysteroid titer remained constant; a decreasing free ecdysteroid titer was accompanieid by increasing titers of both conjugates and acids resulting from the metabolic processes of hydroxylation, oxidation, epimerization, and phosphorylation. The predominant metabolites throughout development were 3-epi-20-hydroxyecdysonoic acid and the phosphate conjugates of 3-epi-20-hydroxyecdysone and 3-epi-20,26-dihydroxyecdysone. The ultimate inactivation of the ecdysteroids of M. sexta during pupal-adult development is possibly mediated by two pairs of metabolically-linked processes, one leading to a 3-epiecdysteroid acid, and the other to 3-epiecdysteroid phosphates.
    Additional Material: 4 Ill.
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  • 2
    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.
    Additional Material: 7 Ill.
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  • 3
    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.
    Additional Material: 10 Ill.
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  • 4
    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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  • 5
    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.
    Additional Material: 9 Ill.
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  • 6
    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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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 1 (1983), S. 25-31 
    ISSN: 0739-4462
    Keywords: honeybees ; pollens ; sterols ; field sites ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Sterols from pollen collected by foraging honeybees, Apis mellifera L, at seven field sites were compared with the sterols of foraging adults and/or prepupae collected from colonies at each site. Invariably, the composition of prepupal sterols was comparable to that found in previous cage studies using chemically defined diets containing various dietary sterols: 24-methyl-enecholesterol was the major sterol; sitosterol and isofucosterol were present in lesser, but significant amounts; and a trace amount of cholesterol was identified in each sample. This occurred even though some of the pollen sterols contained little 24-methylenecholesterol, sitosterol, or isofucosterol and a preponderance of certain other sterols, such as δ7-stigmasten-3β-ol and δ7,24(28)-campestadien-3β-ol in goldenrod and corn pollens, respectively. Thus the selective transfer and utilization of sterols in honeybees that have been demonstrated in cage studies with artificial diets were also shown to occur under field conditions.
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  • 8
    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.
    Additional Material: 3 Ill.
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  • 9
    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.
    Additional Material: 3 Ill.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 2 (1985), S. 385-396 
    ISSN: 0739-4462
    Keywords: ecdysone 20-monooxygenase ; NADPH-cytochrome c reductase ; Manduca sexta ; tobacco hornworm ; midgut ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The dual localization of ecdysone 20-monooxygenase in mitochondria and microsomes of Manduca sexta larval midgut was investigated. Cosubstrate requirements and response to osmolarity of the microsomal ecdysone 20-monooxygenase system were found to be different from those previously reported for the mitochondrial enzyme system. The microsomal monooxygenase utilized NADPH and, less efficiently, NADH as cosubstrates. NADPH and NADH effects were neither additive nor synergistic. NADPH yielded identical activities in isotonic and hypotonic incubations. Mitochondria and microsomes showed no synergistic interaction for ecdysone 20-hydroxylation. After washing of the mitochondria, a large proportion of their ecdysone 20-monooxygenase activity was lost. The extent of the loss was inversely correlated to the concentration of mitochondria in the incubation mixture. The addition of bovine serum albumin to the incubations (2 mg/ml) largely restored the original activities. The microsomal contamination in mitochondrial pellets after each of three successive washings was determined by measuring the activity of a microsomal marker enzyme, NADPH-cytochrome c reductase. At each step of the purification, the ecdysone 20-monooxgenase activity of the mitochondrial preparations far exceeded the activity attributable to the microsomal contamination. These results confirm the existence of two independent ecdysone 20-monooxygenase systems in the midgut of M. sexta larvae.
    Additional Material: 4 Ill.
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