Time filter

Source Type

Prague, Czech Republic

Kratka M.,ASCR Institute of Physics Prague | Babchenko O.,ASCR Institute of Physics Prague | Ukraintsev E.,ASCR Institute of Physics Prague | Vachelova J.,Nuclear Physics Institute of Czech Republic | And 5 more authors.
Diamond and Related Materials | Year: 2016

Diamond is considered as a promising tissue equivalent material in radiation therapies as well as for bioelectronic sensors due to its unique set of properties. These features are combined in this work where effects of gamma irradiation (60Co, up to 300 Gy) on function and stability of microscopic (60×20 μm2) hydrogen-terminated diamond (H-diamond) solution-gated field effect transistors (SG-FETs) are studied. The H-diamond SG-FETs were prepared using 300 nm thin diamond films deposited on glass from methane and hydrogen gas mixture by microwave plasma. Prior to gamma irradiation they were interfaced to proteins (fetal bovine serum) and cells (human sarcoma osteogenic cell line-SAOS2) in cell growth medium. Blank H-diamond SG-FETs did not degrade after the irradiation. With adsorbed proteins and cells they showed specific changes in gate current characteristics (about 100% increase) after the irradiation. These current changes are attributed to modified protein layer and cell morphology on the diamond surface. The presented results establish a first step towards real-time electronicmonitoring of cell growth during the irradiation by therapeutically relevant doses. © 2015 Elsevier B.V. All rights reserved.

Randakova A.,Institute of Physiology CAS | Dolejsi E.,Institute of Physiology CAS | Rudajev V.,Institute of Physiology CAS | Zimcik P.,Institute of Physiology CAS | And 3 more authors.
Pharmacological Research | Year: 2015

We mutated key amino acids of the human variant of the M1 muscarinic receptor that target ligand binding, receptor activation, and receptor-G protein interaction. We compared the effects of these mutations on the action of two atypical M1 functionally preferring agonists (N-desmethylclozapine and xanomeline) and two classical non-selective orthosteric agonists (carbachol and oxotremorine). Mutations of D105 in the orthosteric binding site and mutation of D99 located out of the orthosteric binding site decreased affinity of all tested agonists that was translated as a decrease in potency in accumulation of inositol phosphates and intracellular calcium mobilization. Mutation of D105 decreased the potency of the atypical agonist xanomeline more than that of the classical agonists carbachol and oxotremorine. Mutation of the residues involved in receptor activation (D71) and coupling to G-proteins (R123) completely abolished the functional responses to both classical and atypical agonists. Our data show that both classical and atypical agonists activate hM1 receptors by the same molecular switch that involves D71 in the second transmembrane helix. The principal difference among the studied agonists is rather in the way they interact with D105 in the orthosteric binding site. Furthermore, our data demonstrate a key role of D105 in xanomeline wash-resistant binding and persistent activation of hM1 by wash-resistant xanomeline. © 2015 The Authors. Published by Elsevier Ltd.

Janickova H.,Institute of Physiology CAS | Rudajev V.,Institute of Physiology CAS | Dolejsi E.,Institute of Physiology CAS | Koivisto H.,University of Eastern Finland | And 4 more authors.
Current Alzheimer Research | Year: 2015

Transgenic APPswe/PS1dE9 mice modeling Alzheimer’s disease demonstrate ongoing accumulation of β-amyloid fragments resulting in formation of amyloid plaques that starts at the age of 4-5 months. Buildup of β-amyloid fragments is accompanied by impairment of muscarinic transmission that becomes detectable at this age, well before the appearance of cognitive deficits that manifest around the age of 12 months. We have recently demonstrated that long-term feeding of trangenic mice with specific isocaloric fish oil-based diets improves specific behavioral parameters. Now we report on the influence of short-term feeding (3 weeks) of three isocaloric diets supplemented with Fortasyn (containing fish oil and ingredients supporting membrane renewal), the plant sterol stigmasterol together with fish oil, and stigmasterol alone on markers of cholinergic neurotransmission in the hippocampus of 5-month-old transgenic mice and their wild-type littermates. Transgenic mice fed normal diet demostrated increase in ChAT activity and attenuation of carbachol-stimulated GTP-γ35S binding compared to wild-type mice. None of the tested diets compared to control diet influenced the activities of ChAT, AChE, BuChE, muscarinic receptor density or carbachol-stimulated GTP-γ35S binding in wild-type mice. In contrast, all experimental diets increased the potency of carbachol in stimulating GTP-γ35S binding in trangenic mice to the level found in wild-type animals. Only the Fortasyn diet increased markers of cholinergic synapses in transgenic mice. Our data demonstrate that even short-term feeding of transgenic mice with chow containing specific lipid-based dietary supplements can influence markers of cholinergic synapses and rectify impaired muscarinic signal transduction that develops in transgenic mice. © 2015 Bentham Science Publishers.

Slavikova B.,Czech Institute of Organic Chemistry And Biochemistry | Chodounska H.,Czech Institute of Organic Chemistry And Biochemistry | Nekardova M.,Czech Institute of Organic Chemistry And Biochemistry | Nekardova M.,Charles University | And 6 more authors.
Journal of Medicinal Chemistry | Year: 2016

N-Methyl-d-aspartate receptors (NMDARs) display a critical role in various diseases of the central nervous system. The activity of NMDARs can be modulated by neurosteroids. Herein, we report a structure-activity relationship study for perhydrophenanthrene analogues possessing a framework that mimics the steroidal ring system. This study comprises the design, synthesis, and assessment of the biological activity of a library of perhydrophenanthrene 2-sulfates and 2-hemisuccinates (1-10). Their ability to modulate NMDAR-induced currents was tested on recombinant GluN1/GluN2B receptors. Our results demonstrate that such structural optimization leads to compounds that are inhibitors of NMDARs. Notably, compound 9 (IC50 = 15.6 μM) was assessed as a more potent inhibitor of NMDAR-induced currents than the known endogenous neurosteroid, pregnanolone sulfate (IC50 = 24.6 μM). © 2016 American Chemical Society.

Dolejsi E.,Institute of Physiology CAS | Liraz O.,Tel Aviv University | Rudajev V.,Institute of Physiology CAS | Zimcik P.,Institute of Physiology CAS | And 2 more authors.
Journal of Neurochemistry | Year: 2016

Apolipoprotein E4 (apoE4) is the most prevalent genetic risk factor for Alzheimer's disease. We utilized apoE4-targeted replacement mice (approved by the Tel Aviv University Animal Care Committee) to investigate whether cholinergic dysfunction, which increases during aging and is a hallmark of Alzheimer's disease, is accentuated by apoE4. This revealed that levels of the pre-synaptic cholinergic marker, vesicular acetylcholine transporter in the hippocampus and the corresponding electrically evoked release of acetylcholine, are similar in 4-month-old apoE4 and apolipoprotein E3 (apoE3) mice. Both parameters decrease with age. This decrease is, however, significantly more pronounced in the apoE4 mice. The levels of cholinacetyltransferase (ChAT), acetylcholinesterase (AChE), and butyrylcholinesterase (BuChE) were similar in the hippocampus of young apoE4 and apoE3 mice and decreased during aging. For ChAT, this decrease was similar in the apoE4 and apoE3 mice, whereas it was more pronounced in the apoE4 mice, regarding their corresponding AChE and BuChE levels. The level of muscarinic receptors was higher in the apoE4 than in the apoE3 mice at 4 months and increased to similar levels with age. However, the relative representation of the M1 receptor subtype decreased during aging in apoE4 mice. These results demonstrate impairment of the evoked release of acetylcholine in hippocampus by apoE4 in 12-month-old mice but not in 4-month-old mice. The levels of ChAT and the extent of the M2 receptor-mediated autoregulation of ACh release were similar in the adult mice, suggesting that the apoE4-related inhibition of hippocampal ACh release in these mice is not driven by these parameters. © 2015 The Authors. Journal of Neurochemistry published by John Wiley & Sons Ltd on behalf of International Society for Neurochemistry.

Discover hidden collaborations