Biological responses of selected cell types to the activity of Fucus spiralis Linnaeus i Fucus virsoides algae extracts J.Agardh in vitro

Makroalge iz roda Fucus predstavljaju značajan izvor biološki vrednih komponenti jer su bogate kompleksnim polisaharidima, masnim kiselinama i polifenolima. Cilj ove studije je bio ispitivanje hemijskog sastava, in vitro antikancerskih, antioksidativnih, antidijabetičkih, antimikrobnih svojstava, dihlormetan-metanolnog ekstrakta i tri frakcije algi F. spiralis i F. virsoides kao i sezonskih varijacija u sastavu i aktivnosti između letnjih i jesenjih ekstrakata i frakcija alge F. virsoides. Od osnovnog dihlormetan-metanolnog ekstrakta (1:1) su tečno - tečnom ekstrakcijom dobijene petroletarska, etilacetatna i n-butanolna frakcija. Određen je sadržaj masnih kiselina i ukupnih fenola. DPPH metodom je utvrđena antioksidativna sposobnost ekstrakata i frakcija. MTT testom ispitana je in vitro citotoksičnost ekstrakata i frakcija prema nizu kancerskih ćelijskih linija kao i prema normalnoj ćelijskoj liniji. Distribucija HeLa ćelija (adenokarcinom grlića materice) po fazama ćelijskog ciklusa nakon tretmana ekstraktima i frakcijama, evaluirana je uz pomoć protočne citometrije. Analiziran je tip ćelijske smrti HeLa ćelija nakon primene ekstrakata i frakcija. Antimigracijski i antiangiogeni efekat ekstrakata i frakcija je ispitivan na humanim transformisanim endotelnim ćelijama umbilikalne vene EA.hy926. Ispitan je uticaj ekstrakata i frakcija na inhibiciju enzima α-glukozidza, i time je utvrđena antidijabetička aktivnost. Antimikrobni efekat ekstrakata i frakcija je ispitan mikrodilucionom metodom. Petroletarska frakcija vrste F. spiralis je imala najveći citotoksični efekat prema HeLa ćelijskoj liniji. Jesenji ekstrakt i frakcije vrste F. virsoides su ispoljili jači citotoksični efekat prema kancerskim ćelijskim linijama u poređenju sa letnjim uzorcima. Pokazana je i selektivnost ekstrakata i frakcija u odnosu na normalnu ćelijsku liniju. Rezultati su pokazali razliku u sadržaju masnih kiselina i ukupnih fenola i biološkoj aktivnosti ekstrakata i frakcija dobijenih iz vrste F. virsoides uzorkovanih tokom različitih sezona. Proapototska aktivnost prema HeLa ćelijama je bila izražena nakon tretmana etilacetatnim frakcijama letnjeg uzorka vrste F. virsoides a zatim i vrste F. spiralis. Najizraženiji uticaj na zaustavljane ćelijske migracije EA.hy926 ćelija je iskazala etilacetatna frakcija vrste F. spiralis. Sprečavanje formiranje mreže EA.hy926 ćelija je bilo pod uticajem osnovnih ekstrakata, petroletarske i etilacetane frakcije vrsta F. spiralis i jesenjeg uzorka vrste F. virsoides, dok je samo etilacetatna frakcija letnjeg uzorka vrste F. virsoides iskazala in vitro antiangiogeno dejstvo. Svi ispitani ekstrakti i frakcije su inhibirali aktivnost enzima α-glukozidaze. Ekstrakti i frakcije obe vrste algi su iskazali antimikrobnu i antioksidativnu aktivnost. Ispitivanje biološke aktivnosti ekstrakata i frakcija vrsta F. spiralis i F. virsoides, ukazuje na obećavajući antikancerski i farmakološki potencijal ovih jestivih makroalgi.Macroalgae from the Fucus genus are a valuable source of bioactive components as they are rich in complex polysaccharides, fatty acids, and polyphenols. This study aimed to investigate the chemical composition and in vitro anticancer, antimicrobial, antioxidative and antidiabetic activities of the dichloromethane-methanol extract and three fractions of the Fucus spiralis and Fucus virsoides as well as seasonal variations on algal composition and activity of summer and fall extracts and fractions of Fucus virsoides. Fractions were made from dichloromethane-methanol (1:1) extract of algae using liquid - liquid extraction: petroleum-ether, ethyl-acetate n-butanol. Fatty acid and total phenolic content were obtained. DPPH method was used to determine the antioxidative capacity of extracts and fractions. Extract and fractions were screened for in vitro cytotoxicity by MTT assay against human cancer and normal human cell lines. Cell cycle distribution of the human cervical adenocarcinoma HeLa cells was evaluated using flow cytometry. The type of cell death of HeLa cells was evaluated after treatment with extracts and fractions. Antimigratory and antiangiogenic properties were investigated using scratch and tube formation assays against human endothelium derived permanent EA.hy926 cell line. Inhibition of α-glucosidase assay was used to assess antidiabetic activity. The antimicrobial effect was tested using the microdilution method against bacterial and fungal strains. Petroleum-ether fraction оf F. spiralis exerted the highest cytotoxicity against HeLa cells. Fall extract and fractions of F. virsoides exerted stronger cytotoxic effect against cancer cell lines compared to summer extract and fractions. Cytotoxic activity towards cancer cells was selective compared to normal cells. The seasonal variation in content and biological activity of F. virsoides samples was detected. Results have shown differences in fatty acid and total phenolic content. Proapoptotic activity against HeLa cells was pronounced after treatment with ethyl-acetate fractions of F. virsoides summer sample followed by F. spiralis. Ethyl-acetate fraction of F. spiralis showed the best results in suppressing the migration of EA.hy926 cells. Extracts, petroleum-ether, ethyl-acetate of F. spiralis, and F. virsoides fall samples, as well as ethyl-acetate summer sample of F. virsoides, prevented tube formation of EA.hy926 cells. All tested extract and fractions inhibited α-glucosidase enzymatic activity. Extract and fractions of both species exerted antioxidative and antimicrobial activities. The investigation of biological activities of the extract and fractions obtained from F. spiralis and F. virsoides may suggest a promising anticancer and pharmacological potential of these edible macroalgae

Структурна анализа спирохидантоина са уврнутом амидном везом као антиканцерских агенаса

Five spirohydantoin derivatives were synthesized and analyzed by single crystal X-ray diffraction and DFT calculations.Пет деривата спирохидантоина је синтетисано и проучавано рендгенском структурном анализом и квантно-хемијским прорачунима

Stabilization of Black Locust Flower Extract via Encapsulation Using Alginate and Alginate–Chitosan Microparticles

Black locust flower extract contains various polyphenols and their glucosides contribute to the potential health benefits. After intake of these bioactive compounds and passage through the gastrointestinal tract, their degradation can occur and lead to a loss of biological activity. To overcome this problem, the bioactive compounds should be protected from environmental conditions. This study aimed to encapsulate the black flower extract in the microparticles based on biodegradable polysaccharides, alginate, and chitosan. In the extract, the total antioxidant content was found to be 3.18 ± 0.01 g gallic acid equivalent per 100 g of dry weight. Also, the presence of lipids (16), phenolics (27), organic acids (4), L-aspartic acid derivative, questinol, gibberellic acid, sterol, and saponins (2) was confirmed using the UHPLC–ESI–MS analysis. In vitro assays showed that the extract has weak anti-α-glucosidase activity and moderate antioxidant and cytotoxic activity against the HeLa cell line. The extrusion method with secondary air flow enabled the preparation of microparticles (about 270 μm) encapsulated with extract. An encapsulation efficiency of over 92% was achieved in the alginate and alginate–chitosan microparticles. The swelling study confirmed a lower permeability of alginate–chitosan microparticles compared with alginate microparticles. For both types of microparticles, the release profile of antioxidants in the simulated gastrointestinal fluids at 37 °C followed the Korsmeyer–Peppas model. A lower diffusion coefficient than 0.5 indicated the simple Fick diffusion of antioxidants. The alginate–chitosan microparticles enabled a more sustained release of antioxidants from extract compared to the alginate microparticles. The obtained results indicated an improvement in the antioxidant activity of bioactive compounds from the extract and their protection from degradation in the simulated gastric conditions via encapsulation in the polymer matrixes. Alginate–chitosan showed slightly slower cumulative antioxidant release from microparticles and better antioxidant activity of the extract compared to the alginate system. According to these results, alginate–chitosan microparticles are more suitable for further application in the encapsulation of black locust flower extract. Also, the proposed polymer matrix as a drug delivery system is safe for human use due to its biodegradability and non-toxicity

Cytotoxic and antimicrobial activity of selected Hieracium L. extracts and isolated sesquiterpene lactones

Cytotoxic and antimicrobial activities were investigated for MeOH extracts of flowering aerial parts of Hieracium calophyllum R. Uechtr. (CAL), H. coloriscapum Rohlena & Zahn (COL), H. pseudoschenkii (Rohlena & Zahn) Niketić (PSE), H. valdepilosum Vill. s.l. (VAL) and H. glabratum Willd. (GLA), their two isolated sesquiterpene lactones 8-epiixerisamine A and crepiside E, as well as for CH2Cl2 extract of flowering aerial parts of H. scheppigianum Freyn (SCH). Crepiside E (IC50 = 46.00 μg/mL), COL (IC50 = 152.28 μg/mL) and SCH (IC50 = 148.12 μg/mL) showed the highest cytotoxic activity against HeLa cells in MTT test. The cytotoxic activity against LS174, A549, as well as against normal MRC-5 cells was significantly weaker. In microdilution test, crepiside E and 8-epiixerisamine A exhibited noteworthy antifungal effect against Aspergillus ochraceus, Penicillium funiculosum, Candida albicans and C. krusei (MIC = 0.15 - 0.4 mg/mL, MFC = 0.3 - 0.8 mg/mL). Among the extracts, the best antibacterial activity was shown by SCH and CAL against Pseudomonas aeruginosa (MIC = 1.68 and 2.52 mg/mL, MBC = 3.36 and 5.04 mg/mL), and the highest antifungal activity by SCH and VAL against C. albicans (MIC = 2.48 and 2.52 mg/mL, MFC = 4.96 and 5.04 mg/mL)

Biological responses of selected cell types to the activity of Fucus spiralis Linnaeus i Fucus virsoides algae extracts J.Agardh in vitro

Makroalge iz roda Fucus predstavljaju značajan izvor biološki vrednih komponenti jer su bogate kompleksnim polisaharidima, masnim kiselinama i polifenolima. Cilj ove studije je bio ispitivanje hemijskog sastava, in vitro antikancerskih, antioksidativnih, antidijabetičkih, antimikrobnih svojstava, dihlormetan-metanolnog ekstrakta i tri frakcije algi F. spiralis i F. virsoides kao i sezonskih varijacija u sastavu i aktivnosti između letnjih i jesenjih ekstrakata i frakcija alge F. virsoides. Od osnovnog dihlormetan-metanolnog ekstrakta (1:1) su tečno - tečnom ekstrakcijom dobijene petroletarska, etilacetatna i n-butanolna frakcija. Određen je sadržaj masnih kiselina i ukupnih fenola. DPPH metodom je utvrđena antioksidativna sposobnost ekstrakata i frakcija. MTT testom ispitana je in vitro citotoksičnost ekstrakata i frakcija prema nizu kancerskih ćelijskih linija kao i prema normalnoj ćelijskoj liniji. Distribucija HeLa ćelija (adenokarcinom grlića materice) po fazama ćelijskog ciklusa nakon tretmana ekstraktima i frakcijama, evaluirana je uz pomoć protočne citometrije. Analiziran je tip ćelijske smrti HeLa ćelija nakon primene ekstrakata i frakcija. Antimigracijski i antiangiogeni efekat ekstrakata i frakcija je ispitivan na humanim transformisanim endotelnim ćelijama umbilikalne vene EA.hy926. Ispitan je uticaj ekstrakata i frakcija na inhibiciju enzima α-glukozidza, i time je utvrđena antidijabetička aktivnost. Antimikrobni efekat ekstrakata i frakcija je ispitan mikrodilucionom metodom. Petroletarska frakcija vrste F. spiralis je imala najveći citotoksični efekat prema HeLa ćelijskoj liniji. Jesenji ekstrakt i frakcije vrste F. virsoides su ispoljili jači citotoksični efekat prema kancerskim ćelijskim linijama u poređenju sa letnjim uzorcima. Pokazana je i selektivnost ekstrakata i frakcija u odnosu na normalnu ćelijsku liniju. Rezultati su pokazali razliku u sadržaju masnih kiselina i ukupnih fenola i biološkoj aktivnosti ekstrakata i frakcija dobijenih iz vrste F. virsoides uzorkovanih tokom različitih sezona. Proapototska aktivnost prema HeLa ćelijama je bila izražena nakon tretmana etilacetatnim frakcijama letnjeg uzorka vrste F. virsoides a zatim i vrste F. spiralis. Najizraženiji uticaj na zaustavljane ćelijske migracije EA.hy926 ćelija je iskazala etilacetatna frakcija vrste F. spiralis. Sprečavanje formiranje mreže EA.hy926 ćelija je bilo pod uticajem osnovnih ekstrakata, petroletarske i etilacetane frakcije vrsta F. spiralis i jesenjeg uzorka vrste F. virsoides, dok je samo etilacetatna frakcija letnjeg uzorka vrste F. virsoides iskazala in vitro antiangiogeno dejstvo. Svi ispitani ekstrakti i frakcije su inhibirali aktivnost enzima α-glukozidaze. Ekstrakti i frakcije obe vrste algi su iskazali antimikrobnu i antioksidativnu aktivnost. Ispitivanje biološke aktivnosti ekstrakata i frakcija vrsta F. spiralis i F. virsoides, ukazuje na obećavajući antikancerski i farmakološki potencijal ovih jestivih makroalgi.Macroalgae from the Fucus genus are a valuable source of bioactive components as they are rich in complex polysaccharides, fatty acids, and polyphenols. This study aimed to investigate the chemical composition and in vitro anticancer, antimicrobial, antioxidative and antidiabetic activities of the dichloromethane-methanol extract and three fractions of the Fucus spiralis and Fucus virsoides as well as seasonal variations on algal composition and activity of summer and fall extracts and fractions of Fucus virsoides. Fractions were made from dichloromethane-methanol (1:1) extract of algae using liquid - liquid extraction: petroleum-ether, ethyl-acetate n-butanol. Fatty acid and total phenolic content were obtained. DPPH method was used to determine the antioxidative capacity of extracts and fractions. Extract and fractions were screened for in vitro cytotoxicity by MTT assay against human cancer and normal human cell lines. Cell cycle distribution of the human cervical adenocarcinoma HeLa cells was evaluated using flow cytometry. The type of cell death of HeLa cells was evaluated after treatment with extracts and fractions. Antimigratory and antiangiogenic properties were investigated using scratch and tube formation assays against human endothelium derived permanent EA.hy926 cell line. Inhibition of α-glucosidase assay was used to assess antidiabetic activity. The antimicrobial effect was tested using the microdilution method against bacterial and fungal strains. Petroleum-ether fraction оf F. spiralis exerted the highest cytotoxicity against HeLa cells. Fall extract and fractions of F. virsoides exerted stronger cytotoxic effect against cancer cell lines compared to summer extract and fractions. Cytotoxic activity towards cancer cells was selective compared to normal cells. The seasonal variation in content and biological activity of F. virsoides samples was detected. Results have shown differences in fatty acid and total phenolic content. Proapoptotic activity against HeLa cells was pronounced after treatment with ethyl-acetate fractions of F. virsoides summer sample followed by F. spiralis. Ethyl-acetate fraction of F. spiralis showed the best results in suppressing the migration of EA.hy926 cells. Extracts, petroleum-ether, ethyl-acetate of F. spiralis, and F. virsoides fall samples, as well as ethyl-acetate summer sample of F. virsoides, prevented tube formation of EA.hy926 cells. All tested extract and fractions inhibited α-glucosidase enzymatic activity. Extract and fractions of both species exerted antioxidative and antimicrobial activities. The investigation of biological activities of the extract and fractions obtained from F. spiralis and F. virsoides may suggest a promising anticancer and pharmacological potential of these edible macroalgae

STRUCTURAL ANALYSIS OF TWISTED AMIDES BASED ON THE SPIROHYDANTOIN SCAFFOLD WITH THE ANTICANCER ACTIVITY

Five spirohydantoin derivatives were synthesized and analyzed by single crystal X-ray diffraction and DFT calculations (Figure 1). Compounds 1–3 and 5 crystallize as a syn conformation with respect to the ureic part (N1–C2(=O)–N3) of the hydanoin moiety, while compound 4 crystallize as an anti conformation. All compounds feature a distorted exocyclic amide bond that results from the twisting and pyramidalization of the N3 atom (Figure 2). Compound 2 with τ = 86.9° (97% of theoretical maximum) contains one of the most twisted amide bonds reported so far. The anticancer effects of the investigated compounds were evaluated against selected human cancer cell lines, as well as human normal lung fibroblasts (MRC5) cell lines. It appears that moderately distorted twisted amides with highly lipophilic substituents are the most potent within the series

Stabilization of Black Locust Flower Extract via Encapsulation Using Alginate and Alginate–Chitosan Microparticles

Black locust flower extract contains various polyphenols and their glucosides contribute to the potential health benefits. After intake of these bioactive compounds and passage through the gastrointestinal tract, their degradation can occur and lead to a loss of biological activity. To overcome this problem, the bioactive compounds should be protected from environmental conditions. This study aimed to encapsulate the black flower extract in the microparticles based on biodegradable polysaccharides, alginate, and chitosan. In the extract, the total antioxidant content was found to be 3.18 ± 0.01 g gallic acid equivalent per 100 g of dry weight. Also, the presence of lipids (16), phenolics (27), organic acids (4), L-aspartic acid derivative, questinol, gibberellic acid, sterol, and saponins (2) was confirmed using the UHPLC–ESI–MS analysis. In vitro assays showed that the extract has weak anti-α-glucosidase activity and moderate antioxidant and cytotoxic activity against the HeLa cell line. The extrusion method with secondary air flow enabled the preparation of microparticles (about 270 μm) encapsulated with extract. An encapsulation efficiency of over 92% was achieved in the alginate and alginate–chitosan microparticles. The swelling study confirmed a lower permeability of alginate–chitosan microparticles compared with alginate microparticles. For both types of microparticles, the release profile of antioxidants in the simulated gastrointestinal fluids at 37 °C followed the Korsmeyer–Peppas model. A lower diffusion coefficient than 0.5 indicated the simple Fick diffusion of antioxidants. The alginate–chitosan microparticles enabled a more sustained release of antioxidants from extract compared to the alginate microparticles. The obtained results indicated an improvement in the antioxidant activity of bioactive compounds from the extract and their protection from degradation in the simulated gastric conditions via encapsulation in the polymer matrixes. Alginate–chitosan showed slightly slower cumulative antioxidant release from microparticles and better antioxidant activity of the extract compared to the alginate system. According to these results, alginate–chitosan microparticles are more suitable for further application in the encapsulation of black locust flower extract. Also, the proposed polymer matrix as a drug delivery system is safe for human use due to its biodegradability and non-toxicity