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Toma Tomov, Associate Professor, PhD

Biliana Nikolova, Research Associate, PhD

Marina Suss, Biologist

Yordan Yordanov, Biologist

Aneliya Kostadinova, Research Associate

Nataliya Krasteva, Research Associate,  PhD

Roumiana Tzoneva, Research Associate, PhD

Dencho Gugutkov, Biologist

Victoria Pechlivanova, Biologist

Kameliya Hristova, PhD student

Nikolay Zaekov, PhD student

George Altankov – Professor, Associate member of IBF-BAS

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DNA Transfer Through Membrane

Electroporation, electrofusion and electrotransformatiom. Electrotransfer of macromolecules through lipid membranes. Electrical properties of biological membranes. Electrochemotherapy of tumour cells.

Cell adhesion and reorganization of cytoskeleton after electrotreatment. Influence of low voltage, low frequency electric field on the adhesive properties of nanostructures (tissue engineering).

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Electroporation and electrotransformation methods, patch clamp, fluorescent method for measuring of transmembrane potentials, radioisotope methods, determination of particle electrophoretic mobility, DNA fragmentation, MMT test, FACS analysis.

Cell Adhesion:

1. Colorimetric assays for cell adhesion, growth and viablitiy.
2. Immunofluorescence microscopy
   • Determing the extend of cell spreading and the strength of cell adhesion
   • Visualization of cell cytoskeleton
   • Dynamic of cell receptors upon cell adhesion
3. Enzyme immunoassay
4. Western Blotting
5. Immunoprecipitation
6. Zymography

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Collaborations

1. Prof. E. Peycheva, M. Georgieva, Ph. D., National Centre of Oncology, Sofia.

2. Prof. I. Dozinsky, Centre of Biomedical Engineering, BAS, Sofia

3. Prof. M. P. Rols, CNRS, Toulouse, France.

4. Prof. E. Neumann, Chemical Faculty, University of Bielefeld, Germany.

5. Prof. M. Berger, German Cancer Research Center, Heidelberg, Germany.

6. Prof. G. Altankov, Institute for Bioengineering of  Catalonia, Spain.

7. Prof. T. Groth, Martin-Luter-Univirsitetat, Halle-Wintenberg, Germany.

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International and National Projects

International Projects:

1. French-Bulgarian Laboratory “Vesicles and Membranes” 1997-2002.

2. Biophysical Mechanism of the effect of electromagnetic field on biological and model membrane, EU project-COST 244, 1994-1996.

3-4. Mechanism of electrical permeabilisation, DFG, 436-100, 1993-1998 (2 projects).

5. Mechanism of electroporative DNA and oligo transport through cell membranes and lipid bilayers, DFG 436/113, 1999-2001.

6. Transmembrane transport of anticancer drugs and DNA by electrical pulses, French-Bulgarian project, (CNRS-BAS), 2003-2005, Toulouse, France.

7. Electrotransfer of anticancer drugs and DNA, French-Bulgarian project, (CNRS-BAS), 2006-2007, Toulouse, France.

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National Projects:

8. Electrotransfer of macromolecules, K-31, 1991-1994.

9. Effect of electrical pulses at the interaction and transfer of poly ions through biological membranes, K-411, 1994-1997.

10. Family of apparatus for electrochemotherapy of malignant tumors: laboratory and clinical studies - prototypes and construction- N 5011, 1998-1999.

11. Effect of electric field on macromolecules in biological and model membranes, K-707, 1997-2002.

12. Transmembrane transfer of drugs and DNA by electrical pulses, K-1303, 2003-2006.

13. Exploitation and characterization of nanostrctured biomimetic surfaces  as substrates for cultivating of osteoblasts –DAAD15, 2007.

14. Study of the interaction cell-nanostructured biomaterial (nanofiber nonwoven textile). НТ-4, МОН 2006-2008. 

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Tsoneva I, Nikolova B, Georgieva M, Guenova M, Tomov T, Rols M-P, Berger M R, (2005) Induction of apoptosis by electrotransfer of positively charged proteins as Cytochrom C and Histone H1 into cells. Biochem. Biophys. Acta 1721, 55-64

Berger M, Tsoneva I, Konstantinov S, Eibl H, (2003) Induction of apoptosis by EPC3 is associated with changes in signal molecule expression and location. Ann NY Acad. Sci. 1010, 307-311.

Dimitrov V, Stoimenova M, Tsoneva I, (2002) Electrically induced concentration fluctuations in E. coli suspensions. Colloid and Surfaces, A Physicochem and Eng. Aspects 209, 201-205.

Hristova N, Angelova M, Tsoneva I, (2002) An experimental approach for direct observation of the interaction of polyanions with spingosine-containing giant vesicles. Bioelectrochem. 58, 65-73.

Nikolova B, Dimitrov V, Tsoneva I, (2000) Mechanism of DNA transfer to E. coli by weak alternating electric. J. Balkan Ecol. 3, 48-56.

Tomov T, Tsoneva I, (2000) Are the stainless steel electrodes inert? Bioelectrochem. 51, 207-209.

Angelova M, Tsoneva I, (1999) Interaction of DNA with giant liposomes. Chem. Phys. Lipids. 101, 123-137.

Angelova M, Hristova N, Tsoneva I (1999) DNA -induced endocytosis upon local microinjection to giant unilamellar cationic vesicles. Eur. Biophys. J. 28, 142-150.

Phylipova E, Suss M, Tomov T, Tsoneva I (1997) Effect of external electric fields on the activity of membrane bound enzymes at transmembrane potential equal to zero. C.R. Bulg. Acad. Sci. 50, 105-108.

Hristova N, Tsoneva I, Neumann E (1997) Sphingosine-mediated electroporative DNA transfer through lipid bilayer. FEBS Lett. 415, 81- 86.

Neumann E, Kakorin S, Tsoneva I, Nikolova B, Tomov T (1996) Calcium mediated DNA adsorption to yeast cells and kinetics of cell transformation by electroporation. Biophys. J. 71, 868- 877.

Tomov T, (1995) Quantitative dependence of electroporation on the pulse parameters. Bioelectochem.  Bioenerg. 37, 101-107.

Spassova M, Petrov AG, Tsoneva I, Petkova JI, Neumann E (1994) Dip patch clamp currents suggest electrodifusive transport of the polyelectrolyte DNA through lipid bilayers. Biophys. Chem. 52, 267-274.

Ganeva V, Tsoneva I (1993) Effect of n-alcohols on the electrotransformation and permeability of Saccharomyces cerevisiae. Appl. Microbiol. Biothechnol. 38, 792- 798.

Tsoneva I, Tomov T, Panova I, Doncheva J (1992) Electrofusion in the presence of divalent cations. Bioelectochem. Bioenerg. 27, 37-40.

Tomov T, Tsoneva I (1992) Influence of electrical pulse on the transmembrane potential in yeast. Bioelectrochem. Bioenerg. 27, 33 -36.

Tsoneva I, Panova I, Tomov T, Strahilov D (1990) Effective production of hybridomas secreting monoclonal antibodies against Hc antigen of Salmonella by electrofusion. Bioelectrochem. Bioenerg. 24, 41-49.

Tomov T, Tsoneva I (1989) Changes in the surface charge of cells induced by electrical pulses. Bioelectrochem. Bioenerg. 22, 127-133.

Tomov T, Tsoneva I, Doncheva, J (1988) Electrical stability of erythrocytes in the presence of divalent cations. Biosci Rep. 8, 421-426.

Tomov T, Tsoneva I (1988) Electroinduction of long - lived potential in yeast. Bioelectrochem. Bioenerg. 19, 397-403

Tsoneva I, Doinov P, Dimitrov DS (1989) Electrofusion of fragile mutants of Saccharomyces cerevisiae. FEMS Microbiol. Lett. 60, 61-66.

Tomov T, (1986) Pyronin G as a fluorescent probe for quantitative determination of membrane potential of mitochondria. J. Biochem. Biophys. Methods, 13, 29-38.

Tsoneva I, Zhelev D, Dimitrov DS (1986) Red blood cell dielectrophoresis in axisymmetric field. Cell Biophys. 8, 89-101.

Tsoneva I, Nikolova B, Georgieva M, Guenova M, Tomov T, Rols M-P, Berger M R, (2005) Induction of apoptosis by electrotransfer of positively charged proteins as Cytochrom C and Histone H1 into cells. Biochem. Biophys. Acta 1721, 55-64

Berger M, Tsoneva I, Konstantinov S, Eibl H, (2003) Induction of apoptosis by EPC3 is associated with changes in signal molecule expression and location. Ann NY Acad. Sci. 1010, 307-311.

Dimitrov V, Stoimenova M, Tsoneva I, (2002) Electrically induced concentration fluctuations in E. coli suspensions. Colloid and Surfaces, A Physicochem and Eng. Aspects 209, 201-205.

Hristova N, Angelova M, Tsoneva I, (2002) An experimental approach for direct observation of the interaction of polyanions with spingosine-containing giant vesicles. Bioelectrochem. 58, 65-73.

Nikolova B, Dimitrov V, Tsoneva I, (2000) Mechanism of DNA transfer to E. coli by weak alternating electric. J. Balkan Ecol. 3, 48-56.

Tomov T, Tsoneva I, (2000) Are the stainless steel electrodes inert? Bioelectrochem. 51, 207-209.

Angelova M, Tsoneva I, (1999) Interaction of DNA with giant liposomes. Chem. Phys. Lipids. 101, 123-137.

Angelova M, Hristova N, Tsoneva I (1999) DNA -induced endocytosis upon local microinjection to giant unilamellar cationic vesicles. Eur. Biophys. J. 28, 142-150.

Phylipova E, Suss M, Tomov T, Tsoneva I (1997) Effect of external electric fields on the activity of membrane bound enzymes at transmembrane potential equal to zero. C.R. Bulg. Acad. Sci. 50, 105-108.

Hristova N, Tsoneva I, Neumann E (1997) Sphingosine-mediated electroporative DNA transfer through lipid bilayer. FEBS Lett. 415, 81- 86.

Neumann E, Kakorin S, Tsoneva I, Nikolova B, Tomov T (1996) Calcium mediated DNA adsorption to yeast cells and kinetics of cell transformation by electroporation. Biophys. J. 71, 868- 877.

Tomov T, (1995) Quantitative dependence of electroporation on the pulse parameters. Bioelectochem.  Bioenerg. 37, 101-107.

Spassova M, Petrov AG, Tsoneva I, Petkova JI, Neumann E (1994) Dip patch clamp currents suggest electrodifusive transport of the polyelectrolyte DNA through lipid bilayers. Biophys. Chem. 52, 267-274.

Ganeva V, Tsoneva I (1993) Effect of n-alcohols on the electrotransformation and permeability of Saccharomyces cerevisiae. Appl. Microbiol. Biothechnol. 38, 792- 798.

Tsoneva I, Tomov T, Panova I, Doncheva J (1992) Electrofusion in the presence of divalent cations. Bioelectochem. Bioenerg. 27, 37-40.

Tomov T, Tsoneva I (1992) Influence of electrical pulse on the transmembrane potential in yeast. Bioelectrochem. Bioenerg. 27, 33 -36.

Tsoneva I, Panova I, Tomov T, Strahilov D (1990) Effective production of hybridomas secreting monoclonal antibodies against Hc antigen of Salmonella by electrofusion. Bioelectrochem. Bioenerg. 24, 41-49.

Tomov T, Tsoneva I (1989) Changes in the surface charge of cells induced by electrical pulses. Bioelectrochem. Bioenerg. 22, 127-133.

Tomov T, Tsoneva I, Doncheva, J (1988) Electrical stability of erythrocytes in the presence of divalent cations. Biosci Rep. 8, 421-426.

Tomov T, Tsoneva I (1988) Electroinduction of long - lived potential in yeast. Bioelectrochem. Bioenerg. 19, 397-403

Tsoneva I, Doinov P, Dimitrov DS (1989) Electrofusion of fragile mutants of Saccharomyces cerevisiae. FEMS Microbiol. Lett. 60, 61-66.

Tomov T, (1986) Pyronin G as a fluorescent probe for quantitative determination of membrane potential of mitochondria. J. Biochem. Biophys. Methods, 13, 29-38.

Tsoneva I, Zhelev D, Dimitrov DS (1986) Red blood cell dielectrophoresis in axisymmetric field. Cell Biophys. 8, 89-101.

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Tzoneva R, Seifert B, Albrecht W, Richau K, Lendlein A, Groth T (2007) Poly (ether imide) membranes – studies on the effect of surface modification and protein preadsorption on endothelial cell adhesion, growth and function, Journal of Biomaterials Science: Polymer (in press).

Tzoneva R, Faucheux N, Groth T (2007) Wettability of substrata controls cell-substrate and cell-cell adhesions”, Biochim Biophys Acta 1770(11), 1538-1547.    

Faucheux N, Tzoneva R, Nagel MD, Groth T (2006) The dependence of fibrillar adhesions in human fibroblasts on substratum chemistry”, Biomaterials, 2006, 27, 2, 234-245.   

Vladkova TG, Keranov IL, Dineff PD, Youroukov SY, Avramova IA, Krasteva N, Altankov GP (2005) Plasma based Ar+ beam assisted poly(dimethylsiloxane) surface modification. Nuclear Instruments and Methods in Physics Research, Section B: Beam Interactions with Materials and Atoms 236 (1-4), pp. 552-562.

Krasteva N, Seifert B, Hopp M, Malsch G, Albrecht W, Altankov G,  Groth Th (2005) Membranes for biohybrid liver support: The behaviour of C3A hepatoblastoma cells is dependent on the composition of acrylonitrile copolymers. Journal of Biomaterials Science, Polymer Edition 16 (1), pp. 1-22.

Krasteva N, Seifert B, Albrecht W, Weigel T, Schossig M, Altankov G,  Groth Th (2004) Influence of polymer membrane porosity on C3A hepatoblastoma cell adhesive interaction and function. Biomaterials 25 (13), pp. 2467-2476.

Tzoneva R, Groth T, Altankov G, Paul D (2002) Remodelling of fibrinogen by endothelial cells in dependence on fibronectin matrix assembly. Effect of substratum wettability, Journal of Materials Science: Materials in Medicine 13, 1235-1244.

Groth T, Seifert B, Malsch G, Albrecht W, Paul D, Kostadinova A, Krasteva N, Altankov G (2002) Interaction of human skin fibroblasts with moderate wettable polyacrylonitrile-copolymer membranes. Journal of Biomedical Materials Research 61 (2), pp. 290-300.

Krasteva N, Harms U, Albrecht W, Seifert B, Hopp M, Altankov G, Groth T (2002) Membranes for biohybrid liver support systems - Investigations on hepatocyte attachment, morphology and growth. Biomaterials 23 (12), pp. 2467-2478.

Vladkova T, Krasteva N, Kostadinova A, Altankov G (1999) Preparation of PEG-coated surfaces and a study for their interaction with living cells. Journal of Biomaterials Science, Polymer Edition 10 (6), pp. 609-620.

Groth T, Altankov G, Kostadinova A, Krasteva N, Albrecht W, Paul D (1999) Altered vitronectin receptor ({v) integrin) function in fibroblasts adhering on hydrophobic glass. Journal of Biomedical Materials Research 44 (3), pp.341-351.

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