The use of a barley-based well to define cationic betaglucan to study mammalian cell toxicity associated with interactions with biological structures

Among potential macromolecule-based pharmaceuticals, polycations seem particularly interesting due to their proven antimicrobial properties and use as vectors in gene therapy. This makes an understanding of the mechanisms of these molecules’ interaction with living structures important, so the goal of this paper was to propose and carry out experiments that will allow us to characterize these phenomena. Of particular importance is the question of toxicity of such structures to mammalian cells and, in the work presented here, two lines, normal fibroblasts 3T3-L1 and A549 lung cancer, were used to determine this. In this work, three well-defined cationic derivatives of barley-derived betaglucans obtained in a reaction with glycidyltrimethylammonium chloride (BBGGTMAC) with different degrees of cationization (50, 70, and 100% per one glucose unit) and electrostatic charge were studied. The studies address interactions of these polymers with proteins (bovine serum proteins and BSA), nucleic acids (DNA), glycosaminoglycans (heparin), and biological membranes. The results described in this study make it possible to indicate that toxicity is most strongly influenced by interactions with biological membranes and is closely related to the electrostatic charge of the macromolecule. The presentation of this observation was the goal of this publication. This paper also shows, using fluorescently labeled variants of polymers, the penetration and impact on cell structure (only for the polymer with the highest substitution binding to cell membranes is observed) by using confocal and SEM (for the polymer with the highest degree of substitution, and the appearance of additional structures on the surface of the cell membrane is observed). The labeled polymers are also tools used together with dynamic light scattering and calorimetric titration to study their interaction with other biopolymers. As for the interactions with biological membranes, lipid Langmuir monolayers as model membrane systems were used

Preliminary studies on the mechanism of antifungal activity of new cationic β-glucan derivatives obtained from oats and barley

[Image: see text] New chemical structures with antifungal properties are highly desirable from the point of view of modern pharmaceutical science, especially due to the increasingly widespread instances of drug resistance in the case of these diseases. One way to solve this problem is to use polymeric drugs, widely described as biocidal, positively charged macromolecules. In this work, we present the synthesis of new cationic β-glucan derivatives that show selective antifungal activity and at the same time low toxicity toward animal and human cells. Two β-glucans isolated from oats and barley and modified using glycidyltrimethylammonium chloride were obtained and evaluated for biocidal properties on the cells of mammals and pathogenic fungi and bacteria. These compounds were found to be nontoxic to fibroblast and bacterial cells but showed selective toxicity to certain species of filamentous fungi (Scopulariopsis brevicaulis) and yeasts (Cryptococcus neoformans). The most important aspect of this work is the attempt to explain the mechanisms of action of these compounds by studying their interaction with biological membranes. This was achieved by examining the interactions with model biological membranes representative of given families of microorganisms using Langmuir monolayers. The data obtained partly show correlations between the results for model systems and biological experiments and allow indicating that the selective antifungal activity of cationic β-glucans is related to their interaction with fungal biological membranes and partly lack of such interaction toward cells of other organisms. In addition, the obtained macromolecules were characterized by spectral methods (Fourier transform infrared (FTIR) and (1)H nuclear magnetic resonance (NMR) spectroscopies) to confirm that the desired structure was obtained, and their degree of modification and molecular weights were determined

Isolation of beta glucans from Pleurotus ostreatus and Hordeum vulgare fruiting bodies and synthesis of cationic derivatives from so obtained polymers

Wśród izolowanych oraz badanych związków mających potencjalne działanie terapeutyczne często znajdują się składniki pożywienia. Jednym z takich związków zawartych w owocnikach grzybów oraz zbożach jest β-glukan, polisacharyd, który w wielu badaniach wykazał się właściwościami przeciwwirusowymi, antybakteryjnymi oraz antynowotworowymi. W pracy podjęto próbę izolacji β-glukanu z owocników grzybowych Pleurotus ostreatus oraz Hordeum vulgare L oraz innych potencjalnych źródeł β-glukanu. Dokonano syntezy odpowiednich polikationów i dowiedziono działanie przeciwgrzybicze polikationów zbożowych w stosunku do gatunku grzyba Scopulariopsis brevicaulis. Dodatkowo stwierdzono efekt proproliferacyjny polikationów pochodzenia zbożowego oraz słaby efekt toksyczny polikationu grzybowego w stosunku do mysich komórek fibroblastycznych 3T3-L1, w porównaniu do toksycznego efektu komercyjnie dostępnych leków przeciwgrzybiczych.Food ingredients are often found among isolated and tested compounds with potential therapeutic effects. One of such compounds contained in fruiting bodies of mushrooms and cereals is β-glucan, a polysaccharide, which in many studies has shown antiviral, antibacterial and anti-cancer properties. The study attempts to isolate β-glucan from fungal fruiting bodies of Pleurotus ostreatus and Hordeum vulgare L and other potential sources of β-glucan. The appropriate polycations were synthesized and antifungal activity of cereal polycations was proven against the species of the fungus Scopulariopsis brevicaulis. In addition, the pro-proliferative effect of cereal polycations and a weak toxic effect of the fungal polycation on murine 3T3-L1 fibroblastic cells were found, compared to the toxic effect of commercially available antifungal drugs

Interaction of RNA with polypyridyl ruthenium complexes

Terapia celowana w RNA jest jedną z nowych dróg w terapii przeciwnowotworowej. Powoduje ona m. in. uszkodzenie mitochondrium, które odpowiada za wzrost i przetrwanie komórek nowotworowych. Celem pracy było zbadanie oddziaływań trzech kompleksów polipirydylowych rutenu(II) ([Ru(dip)2bpy]2+,[Ru(dip)3]2+ oraz [Ru(bpy)3]2+, gdzie dip = 4,7-difenylo-1,10-fenantrolina, bpy = 2,2’-bipirydyna), które akumulują się w mitochondium. Oddziaływanie zostało zbadane metodami spektroskopii absorpcyjnej oraz spektrofluorescencji. Wyznaczone zostały stałe oddziaływania kompleksów z RNA, które się zmieniają w następującym szeregu: [Ru(dip)2bpy]2+ > [Ru(dip)3]2+ > [Ru(bpy)3]2+. Kompleks [Ru(dip)2bpy]2+ najsilniej oddziałuje z RNA, co wskazuje że oddziaływanie polipirydylowych kompleksów Ru z biomakromolekułami zależy nie tylko od wielkości ligandów, ale również od ich wzajemnego ułożeniaRNA-based therapy has become a new path in anticancer therapy. It causes among others, mitochondrial damage which is responsible for growth and survival of cancer cells. The aim of this thesis was to study the interaction of three ruthenium(II) polypyridyl complexes ([Ru(dip)2bpy]2+,[Ru(dip)3]2+ and [Ru(bpy)3]2+, where dip = 4,7-diphenyl-1,10-phenanthroline, bpy = 2,2’-bipyridine),which are known to accumulate in mitochondria. The interaction was investigated by UV-visible spectroscopy and fluorescence spectroscopy. The binding constants were determined and can be presented in following order [Ru(dip)2bpy]2+ > [Ru(dip)3]2+ > [Ru(bpy)3]2+. Complex [Ru(dip)2bpy]2+ binds most strongly to RNA which indicates that interaction of Ru complexes with biomacromolecules depends not only on the size of ligands but also their mutual positio