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The mechanisms underlying larval diapause in the wax moth (Galleria mellonella) is one of the most throughly studied aspects. At the low temperature of 18°C, the last instar larvae did not pupate but transferred to 30°C they initiated development and pupation in a circadian manner. Different types of surgical manipulations including head-ligation, nerve cord-severance, implantation of the brain, prothoracic glands, accompanied with ecdysteroid titre measurements indicated that diapausing arrest of larval development at 18°C might be due to the nervous inhibition of their prothoracic glands.
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Chronobiology in Poland

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Stefan Kopec 1888-1941

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Malczewska, M. and Cymborowski, В.: Cold-induced supernumerary moults of Galleria mellonella larvae as bioassay for anti-juvenile hormone compounds. Acta physiol, pol., 1988, 39 (2): 112-120. Application of selected anti-juvenile hormone compounds to freshly moulted and chilled Galleria mellonella last instar larvae inhibits the incidence of supernumerary moultings in a dose-dependent manner. This indicates that the chilling Galleria larvae can be used as a simple and reliable bioassay for testing an anti-juvenile hormone activity of various agents. This bioassay is also very convenient for investigation of the mechanism of actions of these compounds.
A comparison between vitellogenesis in virgin and mated females of Tenebrio molitor showed significant differences at each investigated developmental stage. Yolk protein deposition in oocytes, measured as an increase in their size parameters (length, width, and volume), proceeded much faster and was more efficient in mated females as compared to virgins. In fertilized females the gonadotropic cycle showed a cyclicity with an eight-day period while virgin females finish their vitellogenic stage after the first cycle. These differences were reflected in changes in the rate of protein synthesis in the fat body of females completing vitellogenesis or entering the next oogenetic cycle. In the haemolymph, in addition to a large (158 kDa) and two small (56 kDa and 45 kDa) subunits of vitellogenin, there was an abundance of proteins of 80 kDa and 60 kDa.
In developing Gallería mellonella larvae (reared at 30°C) three proteins of 74/ 76 and 81/82 kDa were identified. They represent a group of storage proteins (LHP proteins). In Galleria larvae, the development of which is arrested by low temperature (18°C)/ accumulation of the 74, 76 and 81/82 kDa proteins was detected in the hemolymph. The synthesis of 74 kDa and 76 kDa proteins started after 24 h, and that of about 80 kDa after 96 h following the transfer of larvae from 30°C to 18°C. 20-Hydroxyecdysone inhibited synthesis of the 74 and 76 kDa proteins in larvae exposed to low temperature. The arrest of development of Galleria larvae is associated with the synthesis and accumulation of storage proteins, and ecdysteroids are involved in these processes.
The male reproductive tract of Lepidoptera is an ideal model for the study of the physiological role of peripheral clocks in insects. The latter are significant in the generation and coordination of rhythmic phenomena which facilitate the initial stages of sperm capacitation. This process requires the maintenance of pH in the upper vas deferens (UVD) aided by, among others, H+-ATPase. Our aim was to determine the potential involvement of carbonic anhydrase (CA) in this process, an enzyme tasked with generating protons subsequently utilized by H+-ATPase to acidify the UVD milieu in S. littoralis, during the time when the lumen of this organ is filled with sperm. We attempted to answer the question whether CA activity can be controlled by the biological oscillator present in the male reproductive tract of the cotton leafworm. Using PAGE zymography, the presence of CA was demonstrated in the UVD wall, but not in the luminal fluid nor in the sperm. Using histochemistry, it was shown that CA is active in the UVD epithelium, and that this activity varies throughout the day and is most likely controlled by an endogenous biological clock. Conversely, the application of CA inhibitors, acetazolamide and sodium thiocyanate, in conjunction with an analysis of H+-ATPase activity in the acidification the UVD environment shows that CA most likely does not play a direct role in the regulation of the pH in this organ.
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