Vasil Atanasov
Sofia University
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Publication
Featured researches published by Vasil Atanasov.
Journal of Inorganic Biochemistry | 2009
Ivayla Pantcheva; Petar Dorkov; Vasil Atanasov; M. Mitewa; Boris Shivachev; Rosica P. Nikolova; William S. Sheldrick
The preparation and structural characterization of a new copper(II) complex of the polyether ionophorous antibiotic sodium monensin A (MonNa) are described. Sodium monensin A binds Cu(II) to produce a heterometallic complex of composition [Cu(MonNa)(2)Cl(2)].H(2)O, 1. The crystallographic data of 1 show that the complex crystallizes in monoclinic space group C2 with Cu(II) ion adopting a distorted square-planar geometry. Copper(II) coordinates two anionic sodium monensin ligands and two chloride anions producing a neutral compound. The sodium ion remains in the inner cavity of the ligand retaining its sixfold coordination with oxygen atoms. Replacement of crystallization water by acetonitrile is observed in the crystal structure of the complex 1. Copper(I) salt of the methyl ester of MonNa, 2, was identified by X-ray crystallography as a side product of the reaction of MonNa with Cu(II). Compound 2, [Me-MonNa][H-MonNa][CuCl(2)]Cl, crystallizes in monoclinic space group C2 with the same coordination pattern of the sodium cation but contains a chlorocuprate(I) counter [CuCl(2)](-), which is linear and not coordinated by sodium monensin A. The antibacterial and antioxidant properties as two independent activities of 1 were studied. Compound 1 is effective against aerobic Gram(+)-microorganisms Bacillus subtilis, Bacillus mycoides and Sarcina lutea. Complex 1 shows SOD-like activity comparable with that of the copper(II) ion.
Biochemistry | 2009
Vasil Atanasov; D. Danchev; M. Mitewa; Svetla Petrova
In the present study, we demonstrate for the first time that the potent neurotoxin vipoxin from the venom of Vipera ammodytes meridionalis exhibits hemolytic and anticoagulant properties. By investigating the effects of phospholipids and calcium ions on hemolysis, we established that the phospholipase A2 (PLA2) enzyme activity is responsible for the hemolytic properties. This was confirmed by chemical modification of the PLA2 active-site histidine residue with p-bromophenacylbromide. Applying different clotting assays, we show that the PLA2 is a weakly anticoagulant enzyme, which affects intrinsic tenase complex by the hydrolysis of procoagulant phospholipids, rather than by nonenzymatic mechanisms (binding to specific coagulation factors). The whole complex—vipoxin—does not affect the coagulation system.
Toxicon | 2012
Vishnya Stoyanova; Radoslav Aleksandrov; Maria Lukarska; Deyan Duhalov; Vasil Atanasov; Svetla Petrova
Vipoxin is a potent postsynaptic heterodimeric neurotoxin isolated from the venom of the Bulgarian snake Vipera ammodytes meridionalis, whose snakebites cause different and strongly manifested pathophysiological effects (neurotoxic, hemolytic, anticoagulant, convulsant, hypotensive, hyperglycemic etc.). The neutralization of snake toxins calls for extensive research through the application of different approaches: antibodies, non-immunologic inhibitors, natural products derived from plants and animals, as well as synthetic drugs. In this study, we applied naive Tomlinson I + J (Cambridge, UK) libraries to obtain recombinant human scFv antibodies against the vipoxins two subunits--basic and toxic phospholipase A₂ (PLA₂) and acidic, non-toxic component. We found that 33 of more than hundred tested clones were positive and recognized vipoxin and its subunits. Enriched scFv-phage samples (1.2 × 10⁹ pfu/ml) were analyzed for their binding (ELISA) and enzyme-inhibiting abilities. Single chain Fv-phage clones--D₁₂, E₃, F₆, D₁₀ and G₅ exhihest binding affinity for the toxic component. Clones A₁, D₁₂ and C₁₂ recognized preferentially vipoxins acidic component. Clones E₃, G₅ and H₄ inhibited the enzymatic activity of both vipoxin and its purified and separated toxic subunit to the highest extent. Six of the selected clones (E₃, G₅, H₄, C₁₂, D₁₀ and A₁₁) inhibited direct hemolytic activity of vipoxin and its pure PLA₂ subunit. The obtained specific scFv antibodies will be used for epitope mapping studies required to shed light on the role of the phospholipase A₂ activity for the vipoxin toxicity and its effective neutralization.
Forensic Science International | 2012
Vasil Atanasov; Silviya Stoykova; A. Runiov; T. Dimitrova; D. Aleksandrova; Stefan Tsakovski; M. Mitewa
Diazepam is one of the mostly used benzodiazepines and it is frequently analyzed in different biological samples, especially blood samples. The diazepam stability in the sample matrices is an important factor regarding reliable data obtaining. The storage is the main factor determining the stability of diazepam in blood samples and it is the object of the study presented. Remaining diazepam amount in spiked whole blood and plasma samples were tested at different storage temperatures, in the absence or presence of sodium fluoride as stabilizer as well as the influence of ethanol on diazepam stability was evaluated. The results of the study indicated that the temperature is the main storage factor affecting diazepam stability. In the fluoride stabilized blood samples the amount of diazepam decreases up to 85% of initial level when stored at -20° C for the period of testing (12 weeks). The presence of low (0.5 g/L) or high (3g/L) ethanol concentrations influences the stability of diazepam at -20 °C. In whole blood samples, the combination of sodium fluoride and ethanol decreases additionally (15-25%) the concentration of the analyte. Freeze-thaw experiments of whole blood samples show about 5-9% decrease in diazepam concentration after the first cycle. The freeze-thaw experiments on plasma samples, containing ethanol and/or fluoride show insignificant decreases of analyte concentration. Further experiments on benzodiazepines stability at different storage conditions or in combination of different factors should be undertaken in forensic toxicology to ensure the data quality, their reliability and reproducibility.
Biochimica et Biophysica Acta | 2011
Konstantin Balashev; Vasil Atanasov; M. Mitewa; Svetla Petrova; Thomas Bjørnholm
In this paper we used AFM as an analytical tool to visualize the degradation of a phospholipid bilayer undergoing hydrolysis of the vipoxins PLA(2). We obtained time series images during the degradation process of supported 1, 2-dipalmitoylphosphatidylcholine (DPPC) bilayers and evaluated the occurrence and the growth rate of the bilayer defects. The special resolution of the AFM images allowed us to measure the area and the perimeter length of these defects and to draw conclusions about the kinetics of the enzyme reaction. Moreover, we also report for some unique characteristics discovered during the vipoxins PLA(2) action. Experimentally for the first time, we observed the appearance and the growth of three-dimensional (3D), crystal-like structures within the formed defects of the degraded bilayer. In an effort to explain their nature, we applied bearing image analysis to estimate the volume of these crystals and we found that their growth rate follows a similar kinetic pattern as the degradation rate of the supported bilayer.
Central European Journal of Chemistry | 2009
Anife Ahmedova; Vasil Atanasov; Petja Marinova; Neyko Stoyanov; M. Mitewa
New 2-acyl-1,3-indandione derivatives, compounds 1–4, were obtained by condensation of 2-acetyl-1,3-indandione with benzaldehyde, thiophene-2-aldehyde, thiophene-3-aldehyde and furane-2-aldehyde, respectively. The structures of the newly synthesized 2-substituted 1,3-indandiones were characterized by means of spectroscopic methods (FT-IR, 1H and 13C NMR, UV-Vis and MS). Based on the obtained results it is suggested that the compounds exist in the exocyclic enolic form. Mass spectral fragmentation paths are also proposed. In order to verify the possibility for tautomerization processes of the newly synthesized compounds their absorption spectra were recorded in various solvents. Furthermore, the complexation properties of the compounds with metal(II) ions were also studied. A series of non-charged complexes with Cu(II), Cd(II), Zn(II), Co(II) and Ni(II) was isolated and analyzed by elemental analyses and IR. The paramagnetic Cu(II) complexes were studied by EPR and distorted, flattened tetrahedral structures are predicted. The other metal complexes show the presence of water molecules, most probably coordinated to the metal ion, thus forming octahedral geometry. Ultimately, the studied properties of the newly synthesized compounds, 1–4, suggest that they may find application as extracting agents for metal ions, rather than as optical sensors.
Analytical Letters | 2009
Vasil Atanasov; Svetla Petrova; M. Mitewa
Abstract A modified isocratic reverse phase high-performance liquid-chromatographic (RP-HPLC) method for phospholipase (PLA) activity assay is developed. Natural lecithins and synthetic phospholipids are used as substrates, and released fatty acids are analyzed after single-phase derivatization (with p-bromophenacyl bromide) at low temperature. The procedure allows simultaneous determination of the total and specific phospholipase activity. This method was successfully applied to snake venom PLA2 activity assay using natural (soybean and egg yolk lecithins) and synthetic (dipalmytoylphosphatydylcholine) substrates for quantitative determination of the enzyme activity.
Toxicon | 2010
I. Marinov; Vasil Atanasov; E. Stankova; D. Duhalov; Svetla Petrova; A. Hubenova
The case report presents a severe coagulopathy in a 56-year-old man following envenomation by the snake (Vipera ammodytes ammodytes) on his left hand. Initially the man was in shock, with an extremely low blood pressure and tachycardia. Local signs included a painful blister formation on the envenomation site. Twenty-four hours later, the man developed acute thrombocytopenia (platelets number 10 x 10(9)/l) and ecchimoses formation on the affected limb and on the left side of his body due to a disseminated intravascular coagulation syndrome, which lasted 13 days and required repeated administration of blood products, antivenin and supportive treatment. The patient was discharged from the hospital after 18 days in a good condition. The case report indicates that the coagulopathy may be a serious life-threatening complication after V. ammodytes ammodytes snakebite.
Central European Journal of Medicine | 2008
Vasil Atanasov; Kamen Kanev; M. Mitewa
Quetiapine fumarate (Seroquel®) is an atypical antipsychotic dibenzothiazepine derivative. Due to its extensive hepatic metabolism and low level of unchanged excretion (< 1%) the routine toxicological drug-screening analyses of urine often leads to false negative results. In the present study, we report that a newly identified metabolite of quetiapine, N-desalkylquetiapine, can be used as an indicative marker of quetiapine-intake in urine using common GC-MS screening procedure. The structure of the mentioned metabolite was solved from the mass-spectrum obtained and the quetiapine presence was proved by consequent HPLC plasma analysis.
Chemico-Biological Interactions | 2013
Vasil Atanasov; Iskra Petrova; Christophor Dishovsky
Organophosphorus compounds (OPC) were developed as warfare nerve agents. They are also widely used as pesticides. The drug therapy of intoxication with OPC includes mainly combination of cholinesterase (ChE) reactivators and cholinolytics. There is no single ChE reactivator having an ability to reactivate sufficiently the inhibited enzyme due to the high variability of chemical structure of the inhibitors. The difficulties in reactivation of ChE activity and slight antidote effect regarding intoxication with some OPC are some of the reasons for continuous efforts to obtain new reactivators of ChE. The aim of the present study was to evaluate the efficacy of some ChE reactivators against OPC intoxication (tabun, paraoxon and dichlorvos) in in vitro experiments and to compare their activity to that known for some currently used oximes (obidoxime, HI-6, 2-PAM). Experiments were carried out using rat brain acetylcholinesterase (AChE). Reactivators showed different activity in the reactivation of rat brain AChE after dichlorvos, paraoxon and tabun inhibition. AChE was easier reactivated after paraoxon treatment. The best effect showed BT-07-4M, obidoxime, TMB-4 and BT-08 from the group of symmetric oximes, and Toxidin, BT-05 and BT-03 from asymmetric compounds. The reactivation of brain AChE inhibited with tabun demonstrated better activity of new compound BT-07-4M, TMB-4 and obidoxime from symmetric oximes, and BT-05 and BT-03 possessing asymmetric structure. All compounds showed low activity toward inhibition of AChE caused by dichlorvos. Comparison of two main structure types (symmetric/asymmetric) showed that the symmetric compounds reactivated better AChE, inhibited with this OPC, than asymmetric ones.