Androgen metabolism in oyster Crassostrea gigas: evidence for 17beta-HSD activities and characterization of an aromatase-like activity inhibited by pharmacological compounds and a marine pollutant.

The annual reproductive cycle of oyster Crassostrea gigas depends on environmental factors, but its endocrine regulations are still unknown. Sexual steroids play important roles at this level in vertebrates, and some estradiol effects have been described in invertebrates such as bivalve mollusks. To question these roles in invertebrates, we studied androgen metabolism in C. gigas. Incubations of tissue homogenates with 14C-steroids such as androstenedione (A), testosterone (T), estrone (E1) and estradiol (E2), followed by TLC and HPLC, provide evidence for 17beta-hydroxysteroid dehydrogenases (17beta-HSDs, conversions of A into T, T into A, E1 into E2 and E2 into E1) and aromatase-like (A into E1) activities. The latter activity was further characterized by tritiated water release assay; it was time- and temperature-dependent. Furthermore, this oyster aromatase-like activity was inhibited by 4-hydroxyandrostenedione (IC(50) 0.456 microM) and by other pharmacological compounds including specific cytochrome P450 inhibitors (MR20494, miconazole) and a marine pollutant (tributyltin).     

Molecular pathogenesis of a new glycogenosis caused by a glycogenin-1 mutation

Glycogenin-1 initiates the glycogen synthesis in skeletal muscle by the autocatalytic formation of a short oligosaccharide at tyrosine 195. Glycogenin-1 catalyzes both the glucose-O-tyrosine linkage and the α1,4 glucosidic bonds linking the glucose molecules in the oligosaccharide. We recently described a patient with glycogen depletion in skeletal muscle as a result of a non-functional glycogenin-1. The patient carried a Thr83Met substitution in glycogenin-1. In this study we have investigated the importance of threonine 83 for the catalytic activity of glycogenin-1. Non-glucosylated glycogenin-1 constructs, with various amino acid substitutions in position 83 and 195, were expressed in a cell-free expression system and autoglucosylated in vitro. The autoglucosylation was analyzed by gel-shift on western blot, incorporation of radiolabeled UDP-(14)C-glucose and nano-liquid chromatography with tandem mass spectrometry (LC/MS/MS). We demonstrate that glycogenin-1 with the Thr83Met substitution is unable to form the glucose-O-tyrosine linkage at tyrosine 195 unless co-expressed with the catalytically active Tyr195Phe glycogenin-1. Our results explain the glycogen depletion in the patient expressing only Thr83Met glycogenin-1 and why heterozygous carriers without clinical symptoms show a small proportion of unglucosylated glycogenin-1.     

The elm leaf beetle α-amylase and its activity relationship with insect feeding

The elm leaf beetle, Xanthogalerucella luteola (Coleoptera: Chrysomelidae) is the most serious pest of elm trees. This pest causes severe damage to elm trees during its growth stages and as a result, in the middle of summer, leaves become skeletised and start to fall down. In this work, biochemical characterisation of digestive α-amylase of this insect and its relationship with insect feeding was investigated. The insect gut was isolated and its α-amylase was extracted and starch (1%) was used as a substrate for the enzyme. Results showed that the enzyme’s optimum temperature and pH was 35?°C and 5.5, respectively. Some ions such as NaCl decreased the enzyme activity whilst MgCl₂ and CaCl₂ increased the enzyme activity. Gut content electrophoresis showed that only one α-amylase is active in this insect species. There was a correlation between the amount of leaf eaten by the insect and the amount of the enzyme activity.     

Aspartic acid introduce the functional amine groups on the surface of superparamagnetic Fe(OH)3@Fe3O4 nanoparticles for efficient immobilization of Penaeus vannamei protease

Bioprocess and Biosystems Engineering - In this study, we synthesized super magnetic Fe(OH)3@Fe3O4 nanoparticles (SPIONs) by the co-precipitation method and introduction of amine groups via...     

The synthesis of 19-norandrostenedione from dehydroepiandrosterone in equine placenta is inhibited by aromatase inhibitors 4-hydroxyandrostenedione and fadrozole

19-Norandrostenedione was synthesized in vitro from dehydroepiandrosterone by explants of equine full-term placenta. The synthesis of 19-norandrostenedione was inhibited by two specific aromatase inhibitors, 4-hydroxyandrostenedione and fadrozole.     

LC–MS/MS characterization of combined glycogenin-1 and glycogenin-2 enzymatic activities reveals their self-glucosylation preferences

Glycogen synthesis is initiated by self-glucosylation of the glycosyltransferases glycogenin-1 and -2 that, in the presence of UDP-glucose, form both the first glucose-O-tyrosine linkage, and then stepwise add a series of α1,4-linked glucoses to a growing chain of variable length. Glycogen-1 and -2 coexist in liver glycogen preparations where the proteins are known to form homodimers, and they also have been shown to interact with each other. In order to study how glycogenin-1 and -2 interactions may influence each other's glucosylations we setup a cell-free expression system for in vitro production and glucosylation of glycogenin-1 and -2 in various combinations, and used a mass spectrometry based workflow for the characterization and quantitation of tryptic glycopeptides originating from glycogenin-1 and -2. The analysis revealed that the self-glucosylation endpoint was the incorporation of 4–8 glucose units on Tyr 195 of glycogenin-1, but only 0–4 glucose units on Tyr-228 of glycogenin-2. The glucosylation of glycogenin-2 was enhanced to 2–4 glucose units by the co-presence of enzymatically active glycogenin-1. Glycogenin-2 was, however, unable to glucosylate inactive glycogenin-1, at least not an enzymatically inactivated Thr83Met glycogenin-1 mutant, recently identified in a patient with severe glycogen depletion.     

Interval aerobic training improves bioenergetics state and mitochondrial dynamics of different brain regions in restraint stressed rats

Molecular Biology Reports - Evidence has validated the prophylactic effects of exercising on different aspects of health. On the opposite side, immobilization may lead to various destructive...     

The effect of Reynolds number on the propulsive efficiency of a biomorphic pulsed-jet underwater vehicle

The effect of Reynolds number on the propulsive efficiency of pulsed-jet propulsion was studied experimentally on a self-propelled, pulsed-jet underwater vehicle, dubbed Robosquid due to the similarity of its propulsion system with squid. Robosquid was tested for jet slug length-to-diameter ratios (L/D) in the range 2-6 and dimensionless frequency (St(L)) in the range 0.2-0.6 in a glycerin-water mixture. Digital particle image velocimetry was used for measuring the impulse and energy of jet pulses from the velocity and vorticity fields of the jet flow to calculate the pulsed-jet propulsive efficiency, and compare it with an equivalent steady jet system. Robosquid's Reynolds number (Re) based on average vehicle velocity and vehicle diameter ranged between 37 and 60. The current results for propulsive efficiency were compared to the previously published results in water where Re ranged between 1300 and 2700. The results showed that the average propulsive efficiency decreased by 26% as the average Re decreased from 2000 to 50 while the ratio of pulsed-jet to steady jet efficiency (η(P)/η(P, ss)) increased up to 0.15 (26%) as the Re decreased over the same range and for similar pulsing conditions. The improved η(P)/η(P, ss) at lower Re suggests that pulsed-jet propulsion can be used as an efficient propulsion system for millimeter-scale propulsion applications. The Re = 37-60 conditions in the present investigation, showed a reduced dependence of η(P) and η(P)/η(P, ss)on L/D compared to higher Re results. This may be due to the lack of clearly observed vortex ring pinch-off as L/D increased for this Re regime.