TY  - THES
AU  - Milošević, Maja S.
PY  - 2014
UR  - http://eteze.bg.ac.rs/application/showtheses?thesesId=3654
UR  - https://fedorabg.bg.ac.rs/fedora/get/o:12456/bdef:Content/download
UR  - http://vbs.rs/scripts/cobiss?command=DISPLAY&base=70036&RID=48111119
UR  - http://nardus.mpn.gov.rs/123456789/6382
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/2700
AB  - ATP i drugi purinski nukleotidi su parakrini signalni molekuli koji vezivanjemza purinske receptore (P1 i P2) na ćelijskoj membrani učestvuju u kontrolikontrakcija i modulaciji imunog odgovora u uterusu sisara. Poremećajikontraktilnosti uterusa mogu imati važnu ulogu u nastanku neplodnosti, dismenorejei prevremenog porođaja. U ovom radu ispitano je prisustvo i kinetičke osobineektoenzima koji hidrolizuju vanćelijski ATP i ADP u uterusu pacova (Wistar albino).Takođe, ispitan je in vitro uticaj teških metala (kadmijuma, žive i bakra) kao izaštitni efekat EDTA i L-cisteina na aktivnost ektoenzima u prisustvu jona metala.Rezultati ove studije pokazali su da hidrolizu vanćelijskog ATP i ADP uuterusu pacova katalizuju enzimi iz familije E-NTPDaza. To potvrđuju rezultati:neosetljivosti na inhibitore drugih membranskih ATPaza (oligomicin, ouabain,teofiln i levamisol); pH optimum za maksimalnu aktivnost enzima na alkalnom pH(8.0-8.5) za oba supstrata; zavisnost enzimske aktivnosti od dvovalentnih jona iinhibicija suraminom. Izmerana aktivnost E-NTPDaza u uterusu je veoma visoka uodnosu na ostala tkiva pacova, što se vidi iz dobijenih vrednosti kinetičkihparametara za hidrolizu ATP (Vmax(app)=3.973±0.280 μmol Pi/min/mg,Km(app)=0.506±0.062 mM), odnosno ADP (Vmax(app)=2.854±0.080 μmol Pi/min/mg,Km(app)=0.639±0.031 mM). Slične Km vrednosti za hidrolizu ATP i ADP, odnospočetnih brzina hidrolize ATP/ADP (1:0.57), Chevillard-ova kriva kompeticije ATPi ADP, kao i inhibicija milimolarnim koncentracijama natrijum-azida (5-10 mM)ukazuju da je E-NTPDaza 1 dominantni enzim koji hidrolizuje vanćelijskenukleotide u uterusu pacova. RT-PCR i Western blot analizom pokazana je genska iproteinska ekspresija NTPDaze 1 i 2 u uterusu pacova.U daljem toku istraživanja ispitan je efekat i mehanizma delovanjadvovalentnih jona kadmijuma, žive(II) i bakra(II) na aktivnost E-NTPDaza iz uterusa pacova...
AB  - ATP and other purine nucleotides are paracrine signaling molecules. Acting viapurine receptors (P1 and P2) on the cell membrane, extracellular nucleotidesparticipate in the control of contraction and modulation of the immune response inmammalian uterus. Disorder of uterine contractility may play an important role in thedevelopment of infertility, dysmenorrhea and premature labor. In this study, weexamined the presence and kinetic properties of ectoenzymes that hydrolyzeextracellular ATP and ADP in rat uterus (Wistar albino). Also, effects of heavy metalions (cadmium, mercury, and copper) on the activity of ectoenzymes from rat uteruswere studied in vitro, in the absence and presence of EDTA and L-cysteine.This study showed that hydrolysis of extracellular ATP and ADP in rat uterusis catalyzed by enzymes classified as the E-NTPDase family. The following resultsconfirmed common properties characteristic for E-NTPDases: insensitivity toinhibitors of other membrane ATPase (oligomycin, ouabain, theophylline andlevamisole), an alkaline pH optimum for both substrates (8.0-8.5), divalent cationsdependence and significant inhibition by suramine. E-NTPDase activity in rat uterusappeared to be very high compared to other rat tissues. The apparent Km values were0.506±0.062 and 0.639±0.031 mM, with a calculated Vmax(app) of 3.973±0.280 and2.854±0.080 μmol Pi/min/mg for ATP and ADP, respectively. According to similarapparent Km values for both substrates, the ATP/ADP hydrolysis ratio (1:0:57),Chevillard competition plot, and inhibition by high milimolar concentrations ofsodium-azide (5-10 mM), E-NTPDase 1 is dominant ATP/ADP hydrolyzing enzymein uterine cell membranes. RT-PCR and Western blot analysis revealed gene andprotein expression of NTPDaze 1 and 2 in rat uterus.In the further course of research, the effects and mechanism of action ofdivalent ions (mercury, cadmium and copper) on the activity of E-NTPDases fromrat uterus were examined...
PB  - Универзитет у Београду, Хемијски факултет
T2  - Универзитет у Београду
T1  - Karakterizacija ektoenzima koji hidrolizuju vanćelijski ATP i ADP u uterusu pacova i in vitro uticaj kadmijuma, žive i bakra na aktivnost enzima
T1  - Characterization of ectoenzymes that hydrolyze extracellular ATP and ADP in rat uterus and in vitro effects of cadmium, mercury and copper on enzyme activity
UR  - https://hdl.handle.net/21.15107/rcub_nardus_6382
ER  - 

@phdthesis{
author = "Milošević, Maja S.",
year = "2014",
abstract = "ATP i drugi purinski nukleotidi su parakrini signalni molekuli koji vezivanjemza purinske receptore (P1 i P2) na ćelijskoj membrani učestvuju u kontrolikontrakcija i modulaciji imunog odgovora u uterusu sisara. Poremećajikontraktilnosti uterusa mogu imati važnu ulogu u nastanku neplodnosti, dismenorejei prevremenog porođaja. U ovom radu ispitano je prisustvo i kinetičke osobineektoenzima koji hidrolizuju vanćelijski ATP i ADP u uterusu pacova (Wistar albino).Takođe, ispitan je in vitro uticaj teških metala (kadmijuma, žive i bakra) kao izaštitni efekat EDTA i L-cisteina na aktivnost ektoenzima u prisustvu jona metala.Rezultati ove studije pokazali su da hidrolizu vanćelijskog ATP i ADP uuterusu pacova katalizuju enzimi iz familije E-NTPDaza. To potvrđuju rezultati:neosetljivosti na inhibitore drugih membranskih ATPaza (oligomicin, ouabain,teofiln i levamisol); pH optimum za maksimalnu aktivnost enzima na alkalnom pH(8.0-8.5) za oba supstrata; zavisnost enzimske aktivnosti od dvovalentnih jona iinhibicija suraminom. Izmerana aktivnost E-NTPDaza u uterusu je veoma visoka uodnosu na ostala tkiva pacova, što se vidi iz dobijenih vrednosti kinetičkihparametara za hidrolizu ATP (Vmax(app)=3.973±0.280 μmol Pi/min/mg,Km(app)=0.506±0.062 mM), odnosno ADP (Vmax(app)=2.854±0.080 μmol Pi/min/mg,Km(app)=0.639±0.031 mM). Slične Km vrednosti za hidrolizu ATP i ADP, odnospočetnih brzina hidrolize ATP/ADP (1:0.57), Chevillard-ova kriva kompeticije ATPi ADP, kao i inhibicija milimolarnim koncentracijama natrijum-azida (5-10 mM)ukazuju da je E-NTPDaza 1 dominantni enzim koji hidrolizuje vanćelijskenukleotide u uterusu pacova. RT-PCR i Western blot analizom pokazana je genska iproteinska ekspresija NTPDaze 1 i 2 u uterusu pacova.U daljem toku istraživanja ispitan je efekat i mehanizma delovanjadvovalentnih jona kadmijuma, žive(II) i bakra(II) na aktivnost E-NTPDaza iz uterusa pacova..., ATP and other purine nucleotides are paracrine signaling molecules. Acting viapurine receptors (P1 and P2) on the cell membrane, extracellular nucleotidesparticipate in the control of contraction and modulation of the immune response inmammalian uterus. Disorder of uterine contractility may play an important role in thedevelopment of infertility, dysmenorrhea and premature labor. In this study, weexamined the presence and kinetic properties of ectoenzymes that hydrolyzeextracellular ATP and ADP in rat uterus (Wistar albino). Also, effects of heavy metalions (cadmium, mercury, and copper) on the activity of ectoenzymes from rat uteruswere studied in vitro, in the absence and presence of EDTA and L-cysteine.This study showed that hydrolysis of extracellular ATP and ADP in rat uterusis catalyzed by enzymes classified as the E-NTPDase family. The following resultsconfirmed common properties characteristic for E-NTPDases: insensitivity toinhibitors of other membrane ATPase (oligomycin, ouabain, theophylline andlevamisole), an alkaline pH optimum for both substrates (8.0-8.5), divalent cationsdependence and significant inhibition by suramine. E-NTPDase activity in rat uterusappeared to be very high compared to other rat tissues. The apparent Km values were0.506±0.062 and 0.639±0.031 mM, with a calculated Vmax(app) of 3.973±0.280 and2.854±0.080 μmol Pi/min/mg for ATP and ADP, respectively. According to similarapparent Km values for both substrates, the ATP/ADP hydrolysis ratio (1:0:57),Chevillard competition plot, and inhibition by high milimolar concentrations ofsodium-azide (5-10 mM), E-NTPDase 1 is dominant ATP/ADP hydrolyzing enzymein uterine cell membranes. RT-PCR and Western blot analysis revealed gene andprotein expression of NTPDaze 1 and 2 in rat uterus.In the further course of research, the effects and mechanism of action ofdivalent ions (mercury, cadmium and copper) on the activity of E-NTPDases fromrat uterus were examined...",
publisher = "Универзитет у Београду, Хемијски факултет",
journal = "Универзитет у Београду",
title = "Karakterizacija ektoenzima koji hidrolizuju vanćelijski ATP i ADP u uterusu pacova i in vitro uticaj kadmijuma, žive i bakra na aktivnost enzima, Characterization of ectoenzymes that hydrolyze extracellular ATP and ADP in rat uterus and in vitro effects of cadmium, mercury and copper on enzyme activity",
url = "https://hdl.handle.net/21.15107/rcub_nardus_6382"
}

Milošević, M. S.. (2014). Karakterizacija ektoenzima koji hidrolizuju vanćelijski ATP i ADP u uterusu pacova i in vitro uticaj kadmijuma, žive i bakra na aktivnost enzima. in Универзитет у Београду
Универзитет у Београду, Хемијски факултет..
https://hdl.handle.net/21.15107/rcub_nardus_6382

Milošević MS. Karakterizacija ektoenzima koji hidrolizuju vanćelijski ATP i ADP u uterusu pacova i in vitro uticaj kadmijuma, žive i bakra na aktivnost enzima. in Универзитет у Београду. 2014;.
https://hdl.handle.net/21.15107/rcub_nardus_6382 .

Milošević, Maja S., "Karakterizacija ektoenzima koji hidrolizuju vanćelijski ATP i ADP u uterusu pacova i in vitro uticaj kadmijuma, žive i bakra na aktivnost enzima" in Универзитет у Београду (2014),
https://hdl.handle.net/21.15107/rcub_nardus_6382 .

TY  - JOUR
AU  - Filipović, Dragana
AU  - Zlatkovic, Jelena
AU  - Pavićević, Ivan D.
AU  - Mandić, Ljuba M.
AU  - Demajo, Miroslav
PY  - 2012
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1540
AB  - The c-Jun NH2-terminal kinases (JNKs) are important stress-responsive kinases. They regulate cellular activities by sequential phosphorylation and activation through a mitogen-activated protein kinase cascade, whereas JNKs activation is altered in response to various stressors. In the present study, we used immunoblotting to assess the effect of 21 day of social isolation as the chronic stressor, either sole and in combination with 2 h of acute immobilization or cold (4A degrees C) stress on circulating corticosterone level and phosphorylation status of p46 (phospho-p46/total p46) and p54 (phospho-p54/total p54) JNK isoforms in the cytosolic and nuclear fraction of the prefrontal cortex and hippocampus of male Wistar rats. Also, the phosphorylation status of JNK nuclear down-stream target c-Jun (p-c-Jun/c-Jun) on Ser63 was examined. Both acute stressors with elevated CORT levels led to increased phosphorylation status of cytosolic p54 JNK isoforms but not p46 JNK isoforms only in the hippocampus and no change in phosphorylation status of c-jun in both brain regions. Chronic isolation with unaltered CORT level and reduced responsiveness to novel acute stressors, led to unchanged or reduced phosphorylation status of p46 and p54 JNK isoforms in both fractions and both brain regions, whereas the decrease of c-Jun phosphorylation status was found only in the prefrontal cortex. Our results suggest that compromised JNKs activation following chronic isolation may lead to interruption of JNK signaling, which could be related with neuropsychiatric disorders such as depression or long-lasting neuronal remodeling.
PB  - Springer Wien, Wien
T2  - Journal of Neural Transmission
T1  - Chronic isolation stress compromises JNK/c-Jun signaling in rat brain
VL  - 119
IS  - 11
SP  - 1275
EP  - 1284
DO  - 10.1007/s00702-012-0776-0
ER  - 

@article{
author = "Filipović, Dragana and Zlatkovic, Jelena and Pavićević, Ivan D. and Mandić, Ljuba M. and Demajo, Miroslav",
year = "2012",
abstract = "The c-Jun NH2-terminal kinases (JNKs) are important stress-responsive kinases. They regulate cellular activities by sequential phosphorylation and activation through a mitogen-activated protein kinase cascade, whereas JNKs activation is altered in response to various stressors. In the present study, we used immunoblotting to assess the effect of 21 day of social isolation as the chronic stressor, either sole and in combination with 2 h of acute immobilization or cold (4A degrees C) stress on circulating corticosterone level and phosphorylation status of p46 (phospho-p46/total p46) and p54 (phospho-p54/total p54) JNK isoforms in the cytosolic and nuclear fraction of the prefrontal cortex and hippocampus of male Wistar rats. Also, the phosphorylation status of JNK nuclear down-stream target c-Jun (p-c-Jun/c-Jun) on Ser63 was examined. Both acute stressors with elevated CORT levels led to increased phosphorylation status of cytosolic p54 JNK isoforms but not p46 JNK isoforms only in the hippocampus and no change in phosphorylation status of c-jun in both brain regions. Chronic isolation with unaltered CORT level and reduced responsiveness to novel acute stressors, led to unchanged or reduced phosphorylation status of p46 and p54 JNK isoforms in both fractions and both brain regions, whereas the decrease of c-Jun phosphorylation status was found only in the prefrontal cortex. Our results suggest that compromised JNKs activation following chronic isolation may lead to interruption of JNK signaling, which could be related with neuropsychiatric disorders such as depression or long-lasting neuronal remodeling.",
publisher = "Springer Wien, Wien",
journal = "Journal of Neural Transmission",
title = "Chronic isolation stress compromises JNK/c-Jun signaling in rat brain",
volume = "119",
number = "11",
pages = "1275-1284",
doi = "10.1007/s00702-012-0776-0"
}

Filipović, D., Zlatkovic, J., Pavićević, I. D., Mandić, L. M.,& Demajo, M.. (2012). Chronic isolation stress compromises JNK/c-Jun signaling in rat brain. in Journal of Neural Transmission
Springer Wien, Wien., 119(11), 1275-1284.
https://doi.org/10.1007/s00702-012-0776-0

Filipović D, Zlatkovic J, Pavićević ID, Mandić LM, Demajo M. Chronic isolation stress compromises JNK/c-Jun signaling in rat brain. in Journal of Neural Transmission. 2012;119(11):1275-1284.
doi:10.1007/s00702-012-0776-0 .

Filipović, Dragana, Zlatkovic, Jelena, Pavićević, Ivan D., Mandić, Ljuba M., Demajo, Miroslav, "Chronic isolation stress compromises JNK/c-Jun signaling in rat brain" in Journal of Neural Transmission, 119, no. 11 (2012):1275-1284,
https://doi.org/10.1007/s00702-012-0776-0 . .