757 research outputs found
Interpretation of bafilomycin, pH neutralizing or protease inhibitor treatments in autophagic flux experiments: novel considerations.
Recent publications showed that the kinase MTOR localizes to lysosomes and its activation depends on amino acids inside the lysosomal lumen, implying that autophagic protein degradation is a positive regulator of MTOR in this setting. Since decreased MTOR activity results in autophagy induction, drug treatments that block autolysosomal degradation (a commonly used technique to estimate autophagic flux) may actually interfere not only with lysosomal breakdown, but also increase autophagosome generation through impaired MTOR signaling
Az autofágiában szereplĹ‘ kinázok Ă©s foszfatázok azonosĂtása in vivo RNSi tesztelĂ©ssel ecetmuslicában = An in vivo RNAi screen of Drosophila kinases and phosphatases involved in autophagy
Az autofágia során a sejt saját citoplazmája kerĂĽl lizoszĂłmális lebontásra Ă©s ĂşjrahasznosĂtásra. Az Ă©hezĂ©s a folyamat nagymĂ©rvű indukciĂłjához vezet. Mivel az autofágia megváltozása kĂĽlönfĂ©le emberi betegsĂ©geket okoz, a folyamat megĂ©rtĂ©se terápiás szempontbĂłl is igen fontos. Munkánkban jĂłl táplált Ă©s Ă©hezĹ‘ muslica lárvák gĂ©nexpressziĂłs mintázatát hasonlĂtottuk össze, kontroll Ă©s autofág mutáns állatokban. A genomszintű transzkripciĂłs változások Ă©s egy kis lĂ©ptĂ©kű RNSi vizsgálatsorozat alapján kĂ©t gĂ©nt választottunk ki rĂ©szletesebb jellemzĂ©sre. A lipid foszfatáz EDTP autofágiában betöltött szerepĂ©t jelenleg is vizsgáljuk. A Rack1 (az aktivált protein kináz C kötĹ‘partnere) expressziĂłja megnĹ‘tt az Ă©hezĂ©s során, Ă©s a gĂ©n hiánya az indukált autofágia gátlását okozta. Az endogĂ©n Rack1 fehĂ©rje jelenlĂ©tĂ©t a korai autofág struktĂşrákban kimutattuk fĂ©ny- Ă©s elektronmikroszkĂłpos szinten is, ami arra utal, hogy ezek kialakulásában játszik szerepet. Érdekes mĂłdon a Rack1 fehĂ©rje a glikogĂ©n szemcsĂ©ken is jelen volt, a glikogĂ©n szintáz kinázzal egyĂĽtt. Ezzel összhangban a Rack1 hiánya sejt-autonĂłm mĂłdon a glikogĂ©n raktárak erĹ‘teljes csökkenĂ©sĂ©hez vezetett az emberi májhoz Ă©s zsĂrszövethez hasonlĂłan működĹ‘ Drosophila lárvális zsĂrtestben. MegfigyelĂ©seink alapján a Rack1 állványfehĂ©rje alapvetĹ‘ szerepet játszik az autofágia korai lĂ©pĂ©seiben Ă©s a glikogĂ©n szintĂ©zis során. | Autophagy delivers cytoplasmic material for lysosomal degradation in eukaryotic cells. Starvation induces high levels of autophagy to promote survival in the lack of nutrients. As alterations in autophagy strongly impact human health and disease, complete understanding of this process is necessary for potential future therapies. In this work, we compared the transcriptional profiles of fed and starved control and autophagy-deficient mutant Drosophila larvae to find novel regulators of autophagy. Based on these transcriptional changes and a small-scale RNAi screen, we chose two genes for detailed studies. We are still investigating how the lipid phosphatase EDTP functions in autophagy. Rack1 (Receptor of activated protein kinase C 1) expression increased during nutrient deprivation, and loss of Rack1 function strongly decreased the autophagic response to starvation. Endogenous Rack1 partially colocalized with Atg8a reporters and early autophagic structures on the ultrastructural level, suggesting its involvement in autophagosome formation. Interestingly, endogenous Rack1 also colocalized with glycogen particles in the larval fat body, and with Shaggy, the Drosophila homolog of Glycogen synthase kinase 3B (GSK-3B). In line with these observations, glycogen stores were decreased in Rack1 mutant cells. These results, for the first time, demonstrate the fundamental role of the scaffold protein Rack1 in autophagosome generation, and also in the formation of glycogen particles
A Bayesian Approach to Identify Bitcoin Users
Bitcoin is a digital currency and electronic payment system operating over a
peer-to-peer network on the Internet. One of its most important properties is
the high level of anonymity it provides for its users. The users are identified
by their Bitcoin addresses, which are random strings in the public records of
transactions, the blockchain. When a user initiates a Bitcoin-transaction, his
Bitcoin client program relays messages to other clients through the Bitcoin
network. Monitoring the propagation of these messages and analyzing them
carefully reveal hidden relations. In this paper, we develop a mathematical
model using a probabilistic approach to link Bitcoin addresses and transactions
to the originator IP address. To utilize our model, we carried out experiments
by installing more than a hundred modified Bitcoin clients distributed in the
network to observe as many messages as possible. During a two month observation
period we were able to identify several thousand Bitcoin clients and bind their
transactions to geographical locations
Genetic variability of anther donor versus spontaneous doubled haploid descendents and colchicine induced doubled haploid sweet pepper (Capsicum annuum L.) lines
Haploid (n) and doubled haploid (DH) plants were developed in anther culture of sweet pepper
(Capsicum annuum L.). Regenerants were analyzed by flow cytometry for haploid (n = 12)
and spontaneous doubled haploid (2n = 24) genomes. Haploid plants were forwarded to
colchicine-treatment for induced doubled haploid (2n·) plant production. Molecular
polymorphism of anther donor plants (2n), the haploid regenerants (n), the spontaneous (2n) and
induced (2n·)-DH plants were analysed by RAPD-, SSR- and ISSR-PCR. The analysis of
anther-donor plants compared to DH-descendents showed an unexpectedly wide range of
molecular polymorphism. Our results suggest that genetic changes occurring during meiotic
recombination is higher than those of occurring during colchicine-induced genomic duplication
Synthetic and steel fibers in prestressed, precast long span beams
Four large-scale, prismatic, T-shaped beams with 19 m span were
produced without stirrups, but 2 with synthetic fibres and 2
with steel fibres. Four point bending tests were made to
simulate the load bearing process of built-in beams. Three point
shear tests were made in the uncracked ends of the beams to
prove the shear resistance of FRC. The behaviour of beams made
of synthetic and steel fibre reinforced concrete was compared.
Finite element calculation was made with the fibre reinforced
concrete and plain concrete as well. The material model was
according to Eurocode and the effect of the fibre was according
to RILEM guideline. The numerical and real test load-deflection
results showed close correlation
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