Browsing by Author "Guncheva M."

Now showing 1 - 7 of 7
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    Biophysical properties and cytotoxicity of feruloylated helix lucorum hemocyanin
    (2020-03-20) Guncheva M.; Idakieva K.; Todinova S.; Stoyanova E.; Yancheva D.
    For the first time Helix lucorum hemocyanin (HlH) has been feruloylated. Two HlH conjugates with 40- and 120- ferulic acid residues were prepared, denoted as FA-HlH-1 and FA-HlH-2. Expectedly, the feruloylation of HlH induced a rearrangement of the protein molecule, a decrease in the α-helical structure at the expense of β-structures was observed. Besides, the FA-HlH conjugates were more prone to aggregation, which is probably due to the stabilization of the partially unfolded protein molecules by non-covalent bonding. Interestingly, the thermal stability of HlH was not affected by the modification. The native and feruloylated HlH were not toxic to normal fibroblasts (BJ cells). We observed a decrease in cell viability of breast cancer MCF-7 cells to about 66% after a 48h exposure to 70 µg/well of FA-HlH-2.
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    Characterization of art materials and degradation processes in the exterior wall paintings of the main church of Rila Monastery, Bulgaria
    (2023-09-01) Stamboliyska B.; Tapanov S.; Kovacheva D.; Atanasova-Vladimirova S.; Ranguelov B.; Yancheva D.; Velcheva E.; Stoyanov S.; Guncheva M.; Fischer D.; Lederer A.
    The present study focused on the characterization of the art materials and the degradation processes in the exterior (exonarthex) mural paintings of the main church of the Nativity of the Virgin in Rila monastery, Bulgaria, which is believed to be the last large-scale example of Eastern Orthodox wall painting. For the first time the art materials used to create a unique and colourful polychrome decoration of the outer gallery and the possible degradation products, caused by atmospheric influence - permanent exposure to open air and moisture - were revealed by a multi-technique approach. The mineral pigments were identified by means of attenuated total reflectance Fourier transform infrared (ATR-FTIR), micro-Raman spectroscopy, scanning electron microscopy energy dispersive X-ray spectroscopy (SEM-EDS) and X-ray diffraction (XRD). The natural yellow, red ochre, and green earth pigments, as well as some synthetic ones such as ultramarine and vermillion, were found in stable condition. Minium and emerald green pigments showed chemical transformations due to adverse environmental conditions which lead to chromatic changes of wall paintings. Black discolouration occurred due to the conversion of orange minium to black plattnerite (PbO2) and white discolouration – due to its transformation to white lead carbon oxide (PbCO3). The copper acetoarsenite (Cu(CH3COO)2.3Cu(AsO2)2) in the emerald green pigment showed partial transformation to arsen-containing mineral phases clinoclase and lindackerite, which fortunately did not affect much the colour appearance. Gypsum and calcium oxalate were found in the majority of the microsamples as decay products. Analysis of the binders by enzyme-linked immunosorbent assay (ELISA) implied the use of the Orthodox Church post-Byzantine egg-tempera technique. The registered Ca metal oxalates in accordance with ELISA results suggested binder chemical degradation induced by external factors. Most of the used painting materials are close to the those found in other Eastern Orthodox Byzantine and post-Byzantine monuments which indicates that the wall painting decoration of the main church of Rila monastery continues the post-Byzantine traditions. On the other hand, the study showed that the exonarthex wall paintings of the main charge of Rila monastery bear some new features as the religious artists supplemented the colourful scheme by emerald green as a new pigment and replaced smalt by the brighter synthetic ultramarine.
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    Ketoprofen-based ionic liquids: Synthesis and interactions with bovine serum albumin
    (2020-01-01) Ossowicz P.; Kardaleva P.; Guncheva M.; Klebeko J.; Świątek E.; Janus E.; Yancheva D.; Angelov I.
    The development of ionic liquids based on active pharmaceutical ingredients (API-ILs) is a possible solution to some of the problems of solid and/or hydrophobic drugs such as low solubility and bioavailability, polymorphism and an alternative route of administration could be suggested as compared to the classical drug. Here, we report for the first time the synthesis and detailed characterization of a series of ILs containing a cation amino acid esters and anion ketoprofen (KETO-ILs). The affinity and the binding mode of the KETO-ILs to bovine serum albumin (BSA) were assessed using fluorescence spectroscopy. All compounds bind in a distance not longer than 6.14 nm to the BSA fluorophores. The estimated binding constants (KA) are in order of 105 L mol-1, which is indicative of strong drug or IL-BSA interactions. With respect to the ketoprofen-BSA system, a stronger affinity of the ILs containing l-LeuOEt, l-ValOBu, and l-ValOEt cation towards BSA is clearly seen. Fourier transformed infrared spectroscopy experiments have shown that all studied compounds induced a rearrangement of the protein molecule upon binding, which is consistent with the suggested static mechanism of BSA fluorescence quenching and formation of complexes between BSA and the drugs. All tested compounds were safe for macrophages.
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    Modulation Effect on Tubulin Polymerization, Cytotoxicity and Antioxidant Activity of 1H-Benzimidazole-2-Yl Hydrazones
    (2023-01-01) Argirova M.; Guncheva M.; Momekov G.; Cherneva E.; Mihaylova R.; Rangelov M.; Todorova N.; Denev P.; Anichina K.; Mavrova A.; Yancheva D.
    1H-benzimidazol-2-yl hydrazones with varying hydroxy and methoxy phenyl moieties were designed. Their effect on tubulin polymerization was evaluated in vitro on porcine tubulin. The compounds elongated the nucleation phase and slowed down the tubulin polymerization comparably to nocodazole. The possible binding modes of the hydrazones with tubulin were explored by molecular docking at the colchicine binding site. The anticancer activity was evaluated against human malignant cell lines MCF-7 and AR-230, as well as against normal fibroblast cells 3T3 and CCL-1. The compounds demonstrated a marked antineoplastic activity in low micromolar concentrations in both screened in vitro tumor models. The most active were the trimethoxy substituted derivative 1i and the positional isomers 1j and 1k, containing hydroxy and methoxy substituents: they showed IC50 similar to the reference podophyllotoxin in both tumor cell lines, accompanied with high selectivity towards the malignantly transformed cells. The compounds exerted moderate to high ability to scavenge peroxyl radicals and certain derivatives—1l containing metha-hydroxy and para-methoxy group, and 1b-e with di/trihydroxy phenyl moiety, revealed HORAC values high or comparable to those of well-known phenolic antioxidants. Thus the 1H-benisimidazol-2-yl hydrazones with hydroxy/methoxy phenyl fragments were recognized as new agents exhibiting promising combined antioxidant and antineoplastic action.
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    Structural, thermal, and storage stability of rapana thomasiana hemocyanin in the presence of cholinium-amino acid-based ionic liquids
    (2021-03-02) Guncheva M.; Idakieva K.; Todinova S.; Yancheva D.; Paunova-Krasteva T.; Ossowicz P.; Janus E.
    Novel biocompatible compounds that stabilize proteins in solution are in demand for biomedical and/or biotechnological applications. Here, we evaluated the effect of six ionic liquids, containing mono-or dicholinium [Chol]1or2 cation and anions of charged amino acids such as lysine [Lys], arginine [Arg], aspartic acid [Asp], or glutamic acid [Glu], on the structure, thermal, and storage stability of the Rapana thomasiana hemocyanin (RtH). RtH is a protein with huge biomedicinal potential due to its therapeutic, drug carrier, and adjuvant properties. Overall, the ionic liquids (ILs) induce changes in the secondary structure of RtH. However, the structure near the Cu-active site seems unaltered and the oxygen-binding capacity of the protein is preserved. The ILs showed weak antibacterial activity when tested against three Gram-negative and three Gram-positive bacterial strains. On the contrary, [Chol][Arg] and [Chol][Lys] exhibited high anti-biofilm activity against E. coli 25213 and S. aureus 29213 strains. In addition, the two ILs were able to protect RtH from chemical and microbiological degradation. Maintained or enhanced thermal stability of RtH was observed in the presence of all ILs tested, except for RtH-[Chol]2 [Glu].
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    SYNTHESIS AND STABILITY OF A RAPANA THOMASIANA HEMOCYANIN CONJUGATED WITH VITAMIN B9
    (2020-01-01) Guncheva M.; Raynova Y.; Idakieva K.; Todinova S.; Yancheva D.
    This is the first report on the synthesis of vitamin B9-Rapana thomasiana hemocyanin conjugates (vit. B9-RtH). Proteins modified with 25 and 150 vitamin B9 residues аre obtained in a coupling reaction between N-hydroxysuccinimide-activated γ -carboxylic groups of vitamin B9 and the amino-groups of N-terminal amino acids and lysines comprising the protein. ATR-FTIR spectroscopy is applied to monitor the changes of the secondary structure upon the modification. The comparison of the secondary structure of the native and the vitamin B9-conjugated RtHs shows that the modification results in an increase of the β-structures in a favor of α-helices and coiled structures. The therrmal stability of the novel preparations is evaluated by differential scanning calorimetry (DSC). Despite the serious changes of the conformation, the vit.-B9-RtH conjugates are characterized by a high thermal stability. It is assumed that the changes of the DSC profiles are due to alternative protein-protein interactions, e.g H-bonding, stacking or hydrophobic interactions with vitamin B9 moieties that are possible in the modified RtH molecules
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    Thermal stability and secondary structure of feruloylated Rapana thomasiana hemocyanin
    (2019-11-01) Guncheva M.; Todinova S.; Yancheva D.; Raynova Y.; Idakieva K.
    This is the first report on the conjugation of a hemocyanin from Rapana thomasiana (RtH) with ferulic acid, which itself exhibits an immunostimulatory activity. In a two-step reaction, 20 to 180 ferulic acid residues were covalently linked to one molecule RtH. The effect of modification of RtH with ferulic acid on the thermal unfolding was evaluated using differential scanning calorimetry (DSC). The DSC curves of the feruloylated-RtHs characterize with an asymmetric shape, which is an indication for the existence of more than one structural unit in the analyzed samples. The protein thermal stability is not affected by the modification; however, the profiles of the DSC curves of the feruloylated-RtH and the native RtH differ, which implies the reorganization in the protein molecule and is in correlation with the secondary structure analyses. The conformational changes in the RtH molecules that are due to the feruloylation were followed in the Amide I region (1600–1700 cm−1) of the ATR-FTIR spectra. In all prepared samples, we observed a rearrangement in protein molecule, decrease in α-helices and coiled structures in favor of the β-structures, and no aggregation in comparison with the native RtH.