The influence of the chlorine atom on the biological activity of 2'-hydroxychalcone in relation to the lipid phase of biological membranes - Anticancer and antimicrobial activity.

The study investigates the effect of the presence of a chlorine atom in the 2'-hydroxychalcone molecule on its interaction with model lipid membranes, in order to discern its potential pharmacological activity. Five chlorine derivatives of 2'-hydroxychalcone were synthesized and evaluated against liposomes composed of POPC and enriched with cationic (DOTAP) or anionic (POPG) lipids. The physicochemical properties of the compounds were initially simulated using SwissAdame software, revealing high lipophilicity (ilogP values: 2.79-2.90). The dynamic light scattering analysis of liposomes showed that chloro chalcones induce minor changes in the diameter of liposomes of different surface charges. Fluorescence quenching assays with a TMA-DPH probe demonstrated the strong ability of the compounds to interact with the lipid bilayer, with varying quenching capacities based on chlorine atom position. FTIR studies indicated alterations in carbonyl, phosphate, and choline groups, suggesting a transition area localization rather than deep penetration into the hydrocarbon chains. Additionally, dipole potential reduction was observed in POPC and POPC-POPG membranes, particularly pronounced by derivatives with a chlorine atom in the B ring. Antibacterial and antibiofilm assays revealed enhanced activity of derivatives with a chlorine atom compared to 2'-hydroxychalcone, especially against Gram-positive bacteria. The MIC and MBIC50 values showed increased efficacy in the presence of chlorine with 3'-5'-dichloro-2'-hydroxychalcone demonstrating optimal antimicrobial and antibiofilm activity. Furthermore, antiproliferative assays against breast cancer cell lines indicated higher activity of B-ring chlorine derivatives, particularly against MDA-MB-231 cells. In general, the presence of a chlorine atom in 2'-hydroxychalcone improves its pharmacological potential, with derivatives showing improved antimicrobial, antibiofilm, and antiproliferative activities, especially against aggressive breast cancer cell lines. These findings underscore the importance of molecular structure in modulating biological activity and highlight chalcones with a chlorine as promising candidates for further drug development studies.

A new class of polyphenolic carbosilane dendrimers binds human serum albumin in a structure-dependent fashion.

The use of dendrimers as drug and nucleic acid delivery systems requires knowledge of their interactions with objects on their way to the target. In the present work, we investigated the interaction of a new class of carbosilane dendrimers functionalized with polyphenolic and caffeic acid residues with human serum albumin, which is the most abundant blood protein. The addition of dendrimers to albumin solution decreased the zeta potential of albumin/dendrimer complexes as compared to free albumin, increased density of the fibrillary form of albumin, shifted fluorescence spectrum towards longer wavelengths, induced quenching of tryptophan fluorescence, and decreased ellipticity of circular dichroism resulting from a reduction in the albumin α-helix for random coil structural form. Isothermal titration calorimetry showed that, on average, one molecule of albumin was bound by 6-10 molecules of dendrimers. The zeta size confirmed the binding of the dendrimers to albumin. The interaction of dendrimers and albumin depended on the number of caffeic acid residues and polyethylene glycol modifications in the dendrimer structure. In conclusion, carbosilane polyphenolic dendrimers interact with human albumin changing its structure and electrical properties. However, the consequences of such interaction for the efficacy and side effects of these dendrimers as drug/nucleic acid delivery system requires further research.

Antibacterial Activity of Plant Polyphenols Belonging to the Tannins against Streptococcus mutans-Potential against Dental Caries.

Dental caries (DC) is the most common oral pathology. The main bacteria responsible for DC is Streptococcus mutans. One of the strategies that can decrease or eliminate the risk of DC development is using compounds that will inhibit both the growth and virulence factors of S. mutans. Tannins are plant polyphenols that have strong antibacterial activity. The purpose of this study was to assess the antibacterial activity of three tannins against S. mutans. In this investigation, microbiological tests (MIC and MBC) and physicochemical techniques like the fluorescence measurements of tannins' interaction with S. mutans cell membrane and membrane proteins, zeta potential, and thermodynamic analyses were used to obtain knowledge about the antibacterial potential of the investigated compounds against S. mutans as well as about the mechanisms associated with antibacterial activity. The obtained results demonstrate that the used compounds exhibit high antibacterial activity against S. mutans. The mechanisms of their antibacterial activity are linked to the strong change in the S. mutans membrane fluidity and potential, and to their interaction with membrane proteins that can result in great disturbance of bacterial physiology and ultimately the inhibition of bacterial growth, triggering their death. Therefore, it can be concluded that the investigated compounds can be potentially used as natural factors in the prevention of dental caries.

Cannabidiol and Cannabigerol Modify the Composition and Physicochemical Properties of Keratinocyte Membranes Exposed to UVA.

The action of UVA radiation (both that derived from solar radiation and that used in the treatment of skin diseases) modifies the function and composition of keratinocyte membranes. Therefore, this study aimed to assess the effects of phytocannabinoids (CBD and CBG), used singly and in combination, on the contents of phospholipids, ceramides, lipid rafts and sialic acid in keratinocyte membranes exposed to UVA radiation, together with their structure and functionality. The phytocannabinoids, especially in combination (CBD+CBG), partially prevented increased levels of phosphatidylinositols and sialic acid from occurring and sphingomyelinase activity after the UVA exposure of keratinocytes. This was accompanied by a reduction in the formation of lipid rafts and malondialdehyde, which correlated with the parameters responsible for the integrity and functionality of the keratinocyte membrane (membrane fluidity and permeability and the activity of transmembrane transporters), compared to UVA-irradiated cells. This suggests that the simultaneous use of two phytocannabinoids may have a protective effect on healthy cells, without significantly reducing the therapeutic effect of UV radiation used to treat skin diseases such as psoriasis.

Antibacterial and Antihemolytic Activity of New Biomaterial Based on Glycyrrhizic Acid and Quercetin (GAQ) against Staphylococcus aureus.

The goal of this study is to obtain and characterize the complex of quercetin with glycyrrhizic acid, which is known to serve as a drug delivery system. Quercetin is a flavonoid with a wide range of biological activities, including an antimicrobial effect. However, quercetin instability and low bioavailability that limits its use in medical practice makes it necessary to look for new nanoformulations of it. The formation of the GAQ complex (2:1) was confirmed by using UV and FT-IR spectroscopies. It was found that the GAQ exhibited antimicrobial and antihemolytical activities against S. aureus bacteria and its main virulent factor-α-hemolysin. The IC50 value for the antihemolytical effect of GAQ was 1.923 ± 0.255 µg/mL. Using a fluorescence method, we also showed that the GAQ bound tightly to the toxin that appears to underlie its antihemolytic activity. In addition, another mechanism of the antihemolytic activity of the GAQ against α-hemolysin was shown, namely, its ability to increase the rigidity of the outer layer of the erythrocyte membrane and thus inhibit the incorporation of α-hemolysin into the target cells, increasing their resistance to the toxin. Both of these effects of GAQ were observed at concentrations below the MIC value for S. aureus growth, indicating the potential of the complex as an antivirulence agent.

Interaction of Rhus typhina Tannin with Lipid Nanoparticles: Implication for the Formulation of a Tannin-Liposome Hybrid Biomaterial with Antibacterial Activity.

Tannins are natural plant origin polyphenols that are promising compounds for pharmacological applications due to their strong and different biological activities, including antibacterial activity. Our previous studies demonstrated that sumac tannin, i.e., 3,6-bis-O-di-O-galloyl-1,2,4-tri-O-galloyl-ß-D-glucose (isolated from Rhus typhina L.), possesses strong antibacterial activity against different bacterial strains. One of the crucial factors of the pharmacological activity of tannins is their ability to interact with biomembranes, which may result in the penetration of these compounds into cells or the realization of their activity on the surface. The aim of the current work was to study the interactions of sumac tannin with liposomes as a simple model of the cellular membrane, which is widely used in studies focused on the explanation of the physicochemical nature of molecule-membrane interactions. Additionally, these lipid nanovesicles are very often investigated as nanocarriers for different types of biologically active molecules, such as antibiotics. In the frame of our study, using differential scanning calorimetry, zeta-potential, and fluorescence analysis, we have shown that 3,6-bis-O-di-O-galloyl-1,2,4-tri-O-galloyl-ß-D-glucose interacts strongly with liposomes and can be encapsulated inside them. A formulated sumac-liposome hybrid nanocomplex demonstrated much stronger antibacterial activity in comparison with pure tannin. Overall, by using the high affinity of sumac tannin to liposomes, new, functional nanobiomaterials with strong antibacterial activity against Gram-positive strains, such as S. aureus, S. epidermitis, and B. cereus, can be formulated.

Antimicrobial Activity of Quercetin, Naringenin and Catechin: Flavonoids Inhibit Staphylococcus aureus-Induced Hemolysis and Modify Membranes of Bacteria and Erythrocytes.

Search for novel antimicrobial agents, including plant-derived flavonoids, and evaluation of the mechanisms of their antibacterial activities are pivotal objectives. The goal of this study was to compare the antihemolytic activity of flavonoids, quercetin, naringenin and catechin against sheep erythrocyte lysis induced by α-hemolysin (αHL) produced by the Staphylococcus aureus strain NCTC 5655. We also sought to investigate the membrane-modifying action of the flavonoids. Lipophilic quercetin, but not naringenin or catechin, effectively inhibited the hemolytic activity of αHL at concentrations (IC50 = 65 ± 5 µM) below minimal inhibitory concentration values for S. aureus growth. Quercetin increased the registered bacterial cell diameter, enhanced the fluidity of the inner and surface regions of bacterial cell membranes and raised the rigidity of the hydrophobic region and the fluidity of the surface region of erythrocyte membranes. Our findings provide evidence that the antibacterial activities of the flavonoids resulted from a disorder in the structural organization of bacterial cell membranes, and the antihemolytic effect of quercetin was related to the effect of the flavonoid on the organization of the erythrocyte membrane, which, in turn, increases the resistance of the target cells (erythrocytes) to αHL and inhibits αHL-induced osmotic hemolysis due to prevention of toxin incorporation into the target membrane. We confirmed that cell membrane disorder could be one of the direct modes of antibacterial action of the flavonoids.

Inhibition of AGEs formation, antioxidative, and cytoprotective activity of Sumac (Rhus typhina L.) tannin under hyperglycemia: molecular and cellular study.

It is well known that accumulation of advanced glycation ends products (AGEs) lead to various diseases such as diabetes and diabetic complications. In this study we showed that hydrolysable tannin from Sumac (Rhus typhina L.)-3,6-bis-O-di-O-galloyl-1,2,4-tri-O-galloyl-ß-D-glucose (C55H40O34) inhibited generation of glycation markers in bovine serum albumin such as AGEs, dityrosine, N'-formylkynurenine and kynurenine under high glucose treatment. This effect was accompanied by stabilization of the protein structure, as was shown using ATR-FT-IR spectroscopy and fluorescence methods. C55H40O34 exhibited also a neuroprotective effect in high glucose-exposed Neuro2A cells suppressing ROS formation and expression of phospho NF-κß and iNOS. At the same time C55H40O34 increased expression of heme oxygenase-1 and NAD(P)H: quinone oxidoreductase and mitochondrial complex I and V activities. Results from this study demonstrates a potent antiglycation activity of C55H40O34 in vitro and indicates its possible therapeutic application in glycation related diseases.

Electrophysiological and spectroscopic investigation of hydrolysable tannins interaction with α-hemolysin of S. aureus.

In this study, using bilayer lipid membrane technique, we report a novel facet of antihemolytic activity of two tannins (1,2,3,4,5-penta-O-galloyl-ß-D-glucose (PGG) and 1,2-di-O-galloyl-4,6-valoneoyl-ß-D-glucose (dGVG)), which consists in inhibiting the formation of α-hemolysin channels and blocking the conductivity of already formed channels. These effects were observed at tannin concentrations well below minimal inhibitory concentration values for S. aureus growth. Using spectroscopic methods, we show that these two tannins differing in molecular structure but having the same number of -OH groups and aromatic rings form firm complexes with hemolysin in aqueous solutions, which may underlie the disruption of its subsequent interaction with the membrane, thus preventing hemolysis of erythrocytes. In all experimental settings, PGG was the more active compound compared to dGVG, that indicates the important role of the flexibility of the tannin molecule in interaction with the toxin. In addition, we found that PGG, but not dGVG, was able to block the release of the toxin by bacterial cells. This toxin is a strong pathogenic factor causing a number of diseases and therefore is considered as a virulence target for treatment of S. aureus infection, so the data obtained suggest that PGG and possibly other tannins of similar structure have therapeutic potential in fighting the virulence of S. aureus.

Biophysical interactions of mixed lipid-polymer nanoparticles incorporating curcumin: Potential as antibacterial agent.

The technology of lipid nanoparticles has a long history in drug delivery, which begins with the discovery of liposomes by Alec D Bangham in the 1960s. Since then, numerous studies have been conducted on these systems, and several nanomedicinal products that utilize them have entered the market, with the latest being the COVID-19 vaccines. Despite their success, many aspects of their biophysical behavior are still under investigation. At the same time, their combination with other classes of biomaterials to create more advanced platforms is a promising endeavor. Herein, we developed mixed lipid-polymer nanoparticles with incorporated curcumin as a drug delivery system for therapy, and we studied its interactions with various biosystems. Initially, the nanoparticle physicochemical properties were investigated, where their size, size distribution, surface charge, morphology, drug incorporation and stability were assessed. The incorporation of the drug molecule was approximately 99.8 % for a formulated amount of 10 % by weight of the total membrane components and stable in due time. The association of the nanoparticles with human serum albumin and the effect that this brings upon their properties was studied by several biophysical techniques, including light scattering, thermal analysis and circular dichroism. As a biocompatibility assessment, interactions with erythrocyte membranes and hemolysis induced by the nanoparticles were also studied, with empty nanoparticles being more toxic than drug-loaded ones at high concentrations. Finally, interactions with bacterial membrane proteins of Staphylococcus aureus and the antibacterial effect of the nanoparticles were evaluated, where the effect of curcumin was improved when incorporated inside the nanoparticles. Overall, the developed mixed nanoparticles are promising candidates for the delivery of curcumin to infectious and other types of diseases.

The Structural Changes in the Membranes of Staphylococcus aureus Caused by Hydrolysable Tannins Witness Their Antibacterial Activity.

Polyphenols, including tannins, are phytochemicals with pronounced antimicrobial properties. We studied the activity of two hydrolysable tannins, (i) gallotannin-1,2,3,4,5-penta-O-galloyl-ß-D-glucose (PGG) and (ii) ellagitannin-1,2-di-O-galloyl-4,6-valoneoyl-ß-D-glucose (dGVG), applied alone and in combination with antibiotics against Staphylococcus aureus strain 8324-4. We also evaluated the effect of these tannins on bacterial membrane integrity and fluidity and studied their interaction with membrane proteins and lipids. A correlation between the antimicrobial activity of the tannins and their membranotropic action depending on the tannin molecular structure has been demonstrated. We found that the antibacterial activity of PGG was stronger than dGVG, which can be associated with its larger flexibility, dipole moment, and hydrophobicity. In addition, we also noted the membrane effects of the tannins observed as an increase in the size of released bacterial membrane vesicles.

Concise synthesis of E/F ring spiroethers from tigogenin. Carbaanalogs of steroidal sapogenins and their biological activity.

A four-step synthesis of five- and six-membered E/F ring spiroethers from tigogenin has been developed. An efficient strategy that features bis-Grignard reaction of dinorcholanic lactone with appropriate bis(bromomagnesio)alkanes followed by acid-mediated spirocyclization was employed to construct a new class of steroid compounds having E and F ring junction as an oxa-carbacyclic system. The synthesized carbaanalogs interact with liposomes and albumin, and also exhibit antibacterial and antifungal activity, demonstrating their pharmacological potential.

Enzymatic synthesis and characterization of aryl iodides of some phenolic acids with enhanced antibacterial properties.

Phenolic acids represent a class of drugs with mild antibacterial properties. We have synthesized iodinated gallic and ferulic acids and together with commercially available iodinated forms of salicylic acids studied their cytotoxicity, bacteriostatic and anti-virulence action. Out of these, iodogallic acid had lowest minimal inhibitory concentration (MIC) against Staphylococcus aureus (MIC = 0.4 mM/118.8 µg/ml). Yet, it had strong effect on erythrocyte membrane lipid ordering and on α-hemolysin secretion by the bacteria at lower non-bacteriostatic and non-cytotoxic concentrations (<0.1 mM). Iodogallic acid formed static complexes with α-hemolysin in solutions (logKb = 4.69 ± 0.07) and inhibited its nano-pore conduction in artificial lipid bilayers (IC50 = 37.9 ± 5.3 µM). These effects of iodogallic acid converged on prevention of hemolysis induced by α-hemolysin (IC50 = 41.5 ± 4.2 µM) and pointed to enhanced and diverse anti-virulence properties of some aryl iodides. The analysis of molecular surface electrostatic charge distribution, molecular hydrophilicity, electronegativity, and dipole moment of studied compounds suggested the importance of the number of hydroxyl groups and their proximity to iodine in anti-virulence activity manifestation. In iodogallic acid, charge redistribution resulted in higher hydrophilicity without concomitant change in overall molecular electronegativity and dipole moment compared to non-iodinated gallic acid. This study shows new directions for the development of antibacterial/antivirulence therapeutics.

Hydrolysable tannins change physicochemical parameters of lipid nano-vesicles and reduce DPPH radical - Experimental studies and quantum chemical analysis.

Tannins belong to plant secondary metabolites exhibiting a wide range of biological activity. One of the important aspects of the realization of the biological effects of tannins is the interaction with lipids of cell membranes. In this work we studied the interaction of two hydrolysable tannins: 1,2,3,4,6-penta-O-galloyl-ß-d-glucose (PGG) and 1,2-di-O-galloyl-4,6-valoneoyl-ß-d-glucose (T1) which had the same number of both aromatic rings (5) and hydroxyl groups (15) but differing in flexibility due to the presence of valoneoyl group in the T1 molecule with DMPC (dimyristoylphosphatidylcholine) lipid nano-vesicles (liposomes). Tannins-liposomes interactions were investigated using fluorescence spectroscopy, differential scanning calorimetry, laser Doppler velocimetry, dynamic light scattering and Fourier Transform Infra-Red spectroscopy. It was shown that more flexible PGG molecules stronger decreased the microviscosity of the liposomal membranes and increased the values of negative zeta potential in comparison with the more rigid T1. Both compounds diminished the phase transition temperature of DMPC membranes, interacted with liposomes via PO groups of head of phospholipids and their hydrophobic regions. These tannins neutralized DPPH free radicals with the stoichiometry of the reaction equal 1:1. The effects of the studied compounds on liposomes were discussed in relation to tannin quantum chemical parameters calculated by molecular modeling.

A Study on the Chemistry and Biological Activity of 26-Sulfur Analogs of Diosgenin: Synthesis of 26-Thiodiosgenin S-Mono- and Dioxides, and Their Alkyl Derivatives.

A chemoselective procedure for MCPBA oxidation of 26-thiodiosgenin to corresponding sulfoxides and sulfone was elaborated. An unusual equilibration of sulfoxides in solution was observed. Moreover, α-alkylation of sulfoxide and sulfone was investigated. Finally, the biological activity of obtained compounds was examined.

Flavonoids modulate liposomal membrane structure, regulate mitochondrial membrane permeability and prevent erythrocyte oxidative damage.

In the present work, we investigated the interaction of flavonoids (quercetin, naringenin and catechin) with cellular and artificial membranes. The flavonoids considerably inhibited membrane lipid peroxidation in rat erythrocytes treated with tert-butyl hydroperoxide (700 µM), and the IC50 values for prevention of this process were equal to 9.7 ± 0.8 µM, 8.8 ± 0.7 µM, and 37.8 ± 4.4 µM in the case of quercetin, catechin and naringenin, respectively, and slightly decreased glutathione oxidation. In isolated rat liver mitochondria, quercetin, catechin and naringenin (10-50 µM) dose-dependently increased the sensitivity to Ca2+ ions - induced mitochondrial permeability transition. Using the probes TMA-DPH and DPH we showed that quercetin rather than catechin and naringenin strongly decreased the microfluidity of the 1,2-dimyristoyl-sn-glycero-3-phosphocholine liposomal membrane bilayer at different depths. On the contrary, using the probe Laurdan we observed that naringenin transfer the bilayer to a more ordered state, whereas quercetin dose-dependently decreased the order of lipid molecule packing and increased hydration in the region of polar head groups. The incorporation of the flavonoids, quercetin and naringenin and not catechin, into the liposomes induced an increase in the zeta potential of the membrane and enlarged the area of the bilayer as well as lowered the temperature and the enthalpy of the membrane phase transition. The effects of the flavonoids were connected with modification of membrane fluidity, packing, stability, electrokinetic properties, size and permeability, prevention of oxidative stress, which depended on the nature of the flavonoid molecule and the nature of the membrane.

Inhibition of interaction between Staphylococcus aureus α-hemolysin and erythrocytes membrane by hydrolysable tannins: structure-related activity study.

The objective of the study was a comparative analysis of the antihemolytic activity against two Staphylococcus aureus strains (8325-4 and NCTC 5655) as well as α-hemolysin and of the membrane modifying action of four hydrolysable tannins with different molecular mass and flexibility: 3,6-bis-O-di-O-galloyl-1,2,4-tri-O-galloyl-ß-D-glucose (T1), 1,2,3,4,5-penta-O-galloyl-ß-D-glucose (T2), 3-O-galloyl-1,2-valoneoyl-ß-D-glucose (T3) and 1,2-di-O-galloyl-4,6-valoneoyl-ß-D-glucose (T4). We showed that all the compounds studied manifested antihemolytic effects in the range of 5-50 µM concentrations. However, the degree of the reduction of hemolysis by the investigated tannins was not uniform. A valoneoyl group-containing compounds (T3 and T4) were less active. Inhibition of the hemolysis induced by α-hemolysin was also noticed on preincubated with the tannins and subsequently washed erythrocytes. In this case the efficiency again depended on the tannin structure and could be represented by the following order: T1 > T2 > T4 > T3. We also found a relationship between the degree of antihemolytic activity of the tannins studied and their capacity to increase the ordering parameter of the erythrocyte membrane outer layer and to change zeta potential. Overall, our study showed a potential of the T1 and T2 tannins as anti-virulence agents. The results of this study using tannins with different combinations of molecular mass and flexibility shed additional light on the role of tannin structure in activity manifestation.

Spectroscopic, Zeta-potential and Surface Plasmon Resonance analysis of interaction between potential anti-HIV tannins with different flexibility and human serum albumin.

Tannins belong to secondary metabolites of plants that exhibit a variety of biological activities, including antiviral one. In this research, we studied the interaction of human serum albumin (HSA) with two ellagitannins: 2,4-valoneoyl-3,6-hexahydroxydiphenoyl-ß-d-glucose (T1) and 1,2-di-O-galloyl-3,6-valoneoyl-ß-d-glucose (T2) from Euphorbia species having antiviral potential against HIV and differing in molecular flexibility due to the presence of valoneoyl- and hexahydroxydiphenoyl groups. A fluorescence analysis demonstrated that the tannins studied strongly interacted with HSA and quenched tryptophan (Trp) fluorescence in the range of 0.25-4 µM. The quenching occurred by a static mechanism. The logKb for more flexible T2 was generally higher in comparison with stiffer T1 (4.94 ±â€¯0.82 vs. 4.12 ±â€¯0.31 and 4.94 ±â€¯0.53 vs. 4.07 ±â€¯0.45 for 296 K and 303 K respectively). The difference was also in the nature of the forces participating in the interaction with HSA. The stiff T1 reacted with HSA via hydrophobic forces, whereas the flexible T2 interacted with the protein by van der Waals forces and hydrogen bonds. The nature of the bonds was also confirmed by a study of the hydrophobicity of the compounds. Zeta-potential measurements showed slightly modifications of albumin electric charge but without significant changes in the surface structure of protein. Surface Plasmon Resonance imaging (SPRi) revealed that the used tannins fully saturated a 3 ng/mL solution of albumin at the concentrations of above 15 ng/mL. Our experiments clearly showed that the tannins used formed complexes with HSA and that the flexibility of the tannins was an important factor determining their interaction with the protein.

Ferutinin Induces Membrane Depolarization, Permeability Transition Pore Formation, and Respiration Uncoupling in Isolated Rat Liver Mitochondria by Stimulation of Ca2+-Permeability.

It is well known that the terpenoid ferutinin (4-oxy-6-(4-oxybenzoyloxy) dauc-8,9-en), isolated from the plant Ferula tenuisecta, considerably increases the permeability of artificial and cellular membranes to Ca2+-ions and produces apoptotic cell death in different cell lines in a mitochondria-dependent manner. The present study was designed for further evaluation of the mechanism(s) of mitochondrial effects of ferutinin using isolated rat liver mitochondria. Our findings provide evidence for ferutinin at concentrations of 5-27 µM to decrease state 3 respiration and the acceptor control ratio in the case of glutamate/malate as substrates. Ferutinin alone (10-60 µM) also dose-dependently dissipated membrane potential. In the presence of Ca2+-ions, ferutinin (10-60 µM) induced considerable depolarization of the inner mitochondrial membrane, which was partially inhibited by EGTA, and permeability transition pore formation, which was diminished partly by cyclosporin A, and did not influence markedly the effect of Ca2+ on mitochondrial respiration. Ruthenium Red, a specific inhibitor of mitochondrial calcium uniporter, completely inhibited Ca2+-induced mitochondria swelling and membrane depolarization, but did not affect markedly the stimulation of these Ca2+-dependent processes by ferutinin. We concluded that the mitochondrial effects of ferutinin might be primarily induced by stimulation of mitochondrial membrane Ca2+-permeability, but other mechanisms, such as driving of univalent cations, might be involved.

Interaction of α-synuclein with Rhus typhina tannin - Implication for Parkinson's disease.

The etiology of Parkinson's disease (PD) relates to α-synuclein, a small protein with the ability to aggregate and form Lewy bodies. One of its prevention strategies is inhibition of α-synuclein oligomerization. We have investigated the interaction of α-synuclein and human serum albumin with 3,6-bis-О-di-О-galloyl-1,2,4-tri-О-galloyl-ß-d-glucose (a tannin isolated from the plant Rhus typhina). Using fluorescence spectroscopy method we found that this tannin interacts strongly with α-synuclein forming complexes. Circular dichroism analysis showed a time-dependent inhibition of α-synuclein aggregation in the presence of the tannin. On the other hand, 3,6-bis-О-di-О-galloyl-1,2,4-tri-О-galloyl-ß-d-glucose had a much stronger interaction with human serum albumin than α-synuclein. The calculated binding constant for tannin-protein interaction was considerably higher for albumin than α-synuclein. This tannin interacted with albumin through a "sphere of action" mechanism. The results lead to the conclusion that 3,6-bis-О-di-О-galloyl-1,2,4-tri-О-galloyl-ß-d-glucose is a potent preventive compound against Parkinson's disease. However, this tannin interacts very strongly with human serum albumin, significantly reducing the bioavailability of this compound.