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Human ADA2-specific IgY antibodies from hen eggs with tolerance to human IgG
E. Yu. Sarkisyan1, H. A. Haroutyunyan2, Ye. G. Sargisova1*
1H. Buniatian Institute of Biochemistry of National Academy of Sciences of Armenia, Yerevan;
2Laboratory of Biochemical and Biophysical Investigations,
Scientific-Research Centre, Yerevan State Medical University after Mkhitar Heratsi, Armenia;
*e-mail: lizasar2001@yahoo.com
Received: 22 April 2025; Revised: 09 July 2025;
Accepted: September 2025; Available on-line: 2025
Purification of human plasma adenosine deaminase 2 (ADA2) is challenging due to its low physiological concentration and co-purification of human immunoglobulin G (IgG), which has similar physicochemical properties. This study describes a novel approach exploiting avian immune tolerance to human IgG to selectively produce antibodies against native and active human ADA2. Chickens were rendered tolerant to human IgG, enabling subsequent immunization with a preparation containing both ADA2 and human IgG. This strategy successfully elicited a specific immune response, yielding antibodies exclusively to ADA2. These anti-ADA2 antibodies (IgY), purified from egg yolks, were subsequently employed in an affinity column. This immunopurification method allowed the isolation of an electrophoretically homogeneous preparation of human ADA2. This study demonstrates the utility of inducing immune tolerance to human IgG in chickens.
Influence of lipopolysaccharides of Escherichia coli on the protease activity of several Bacillus strains
L. D. Varbanets1, O. S. Brovarska1, O. V. Gudzenko1*,
K. G. Garkava2, A. R. Makarenko2
1Institute of Microbiology and Virology named after D. K. Zabolotny,
National Academy of Sciences of Ukraine, Kyiv, Ukraine;
2National Technical University of Ukraine
“Igor Sykorsky Kyiv Polytechnic Institute”, Kyiv, Ukraine;
*e-mail: alena.gudzenko81@gmail.com
Received: 07 July 2025; Revised: 23 August 2025;
Accepted: 12 September 2025; Available on-line: 17 September 2025
We have previously shown that lipopolysaccharides (LPS) of a number of strains of the phytopathogenic species Pantoea agglomerans are capable of increasing the activity of Bacillus peptidases with fibrinolytic, elastase and collagenase activities by 2-4 times. The aim of this work was to investigate the effect of isolated intracellular LPS1 and extracellular LPS2 of Escherichia coli on the activity of purified bacilli proteases with elastase and fibrinogenolytic activity. It was shown that both LPS2 and LPS1 of E. coli 23 can increase the elastase activity of Bacillus sp. IMV B-7883 by 600 and 416% respectively. Both LPS are able to increase fibrinogenolytic activity in all studied Bacillus strains, but its greatest stimulation (200%) was observed under the action of LPS2 of Bacillus sp. L9.
Cytochrome P450 dependent free radical processes in the liver microsomes of rats administered diethyl phthalate
О. V. Kеtsa*, М. М. Маrchenko
Educational and Scientific Institute of Biology, Chemistry and Natural Resources,
Yuriy Fedkovych Chernivtsi National University, Chernivtsi, Ukraine;
*e-mail: o.ketsa@chnu.edu.ua
Received: 28 May 2025; Revised: 03 July 2025;
Accepted: 12 September 2025; Available on-line: 17 September2025
Diethyl phthalate (DEP) is widely used as a plasticizer and aromatic additive in various consumer products. Biotransformation of this xenobiotic occurs through the cytochrome P450 (CYP) -hydroxylating system, the catalytic cycle of which is accompanied by ROS generation in uncoupling reactions. The present study investigated the effects of DEP administration on the CYP-dependent ROS generation and lipid peroxidation in the rat liver microsomes. The experiment was conducted on three groups of purebred white rats: control (intact animals); rats orally administered with DEP at a dose of 2.5 or 5.4 mg/kg b.w per day for 21 days. CYP-mediated ROS generation was initiated by adding 0.24 μmol/l NADPH to the incubation mixture. It was found that daily administration of DEP at a dose of 2.5 mg/kg led to an increase in the rate of O2•– formation, H2O2 content, and intensification of lipid peroxidation in the liver microsomes only on the 21st day of the experiment. In contrast, administration of DEP at a dose of 5.4 mg/kg resulted in increased content of primary, secondary and final lipid peroxidation products as early as on the 14th day of xenobiotic exposure, indicating a dose- and time-dependent effect of DEP on the oxidative stress intensity in liver microsomes.
Thiacalix[4]arene С-1193 – a promising inhibitor of the sodium pump in the uterine smooth muscle cells
O. V. Maliuk1*, T. O. Veklich1, O. V. Tsymbalyuk2, O. V. Bevza1,
S. O. Cherenok3, A. I. Selikhova3, V. I. Kalchenko3, S. O. Kosterin1
1Palladin Institute of Biochemistry, National Academy of Sciences
of Ukraine, Kyiv, Ukraine;
*e-mail: sanya2000ua@gmail.com;
2Educational and Scientific Institute of High Technologies,
Taras Shevchenko National University of Kyiv, Kyiv, Ukraine;
3Institute of Organic Chemistry, National Academy of Sciences of Ukraine, Kyiv, Ukraine
Received: 29 May 2025; Revised: 18 July 2025;
Accepted: 12 September 2025; Available on-line: 17 September2025
Thiacalix[4]arene C-1193 (25,27-dibutoxythiacalix[4]arene-bis-hydroxymethylphosphonic acid) was shown to inhibit the activity of Na+,K+-ATPase with a high efficiency (І0.5 = 42.1 ± 0.6 nM) with no effect on the activity of Mg2+-ATPase, Са2+-ATPase and Са2+,Mg2+-ATPase in the plasma membrane fraction of rat uterine smooth muscle cells. The kinetic regularities of the C-1193 inhibitory effect on Na+,K+-ATPase activity were investigated. It was demonstrated that C-1193 increased the enzyme activation constant by Na+ but not by K+ ions. The contractile activity of the rat uterine horns was investigated by tenzometric methods with the use of longitudinal uterine smooth muscle strips with intact endometrium. С-1193 induced a considerable increase in the amplitude of the acetylcholine-induced contractions as well as the maximal velocity of the contraction and relaxation phases. No effect of С-1193 on contractive activity induced by the selective agonist of М3-cholinoreceptors cevimeline was observed. The results of computer simulation showed that С-1193inhibitory effect must be related to the cooperative action of methylene bisphosphonate fragments on the upper rim of the calixarene platform, and the linker sulfur atoms of calixarene “cup” on the Na+,K+-ATPase macrostructure.
ABTS oxidation reaction as a model of cytochrome c-driven electron transfer
F. Gudratova, A. Aliyeva, S. Mahmudova, K. Gasimov, T. Yusifov*
Institute of Biophysics, Ministry of Science and Education
of the Republic of Azerbaijan, Baku;
*e-mail: tjussifo@ucla.edu
Received: 23 May 2025; Revised: 24 July 2025;
Accepted: 12 September 2025; Available on-line: 17 September2025
Cytochrome c, as an electron carrier within the mitochondria, plays a crucial role in the electron transport chain. To meet the demand for rapid methods that assess the electron transport properties of cytochrome c, we used the electron donor 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonate) (ABTS) as a substrate and suitable spectrophotometric reporter of cytochrome c peroxidase-like activity. ABTS and cytochrome c from bovine were purchased from Sigma-Aldrich Inc. The time course of the cytochrome c-driven ABTS oxidation reaction was studied using H2O2 as a second substrate. It was demonstrated that CytC addition is a prerequisite for the transfer of electrons from ABTS to H2O2. The reaction kinetic analysis with determination of Vmax, Km, kcat, and kcat/Km values for both substrates was performed. Our results demonstrate that the cytochrome c-catalyzed ABTS oxidation reaction can be effectively employed as a model for studying the functional role of cytochrome c in various conditions.