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      Chemical and Biological Characteristics of Propolis from Apis Mellifera Caucasica from The Ardahan and Erzurum Provinces of Turkey: A Comparative Study Translated title: Usporedba kemijskih i bioloških značajki propolisa pčelinje pasmine Apis mellifera caucasica iz turskih provincija Ardahana i Erzuruma

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          Abstract

          The aim of this study was to compare the biological activities of ethanolic propolis extracts of Apis mellifera caucasica obtained from Ardahan and Erzurum provinces of Turkey. Samples were tested for antioxidant, anticytotoxic, anticarcinogenic, antibacterial, and antifungal potentials using different techniques. Propolis samples from the two provinces had different mineral and organic compositions related to their geographical origin. The ferric reducing antioxidant power (FRAP) test showed superiority of Ardahan propolis over the Erzurum. Regardless of origin and the presence of mitomycin C in the culture medium, propolis enhanced human peripheral lymphocyte viability, which depended on the duration and propolis concentration. Antiperoxidative activity on MCF-7 breast cancer cells was concentration-dependent. Erzurum propolis showed the highest anticarcinogenic activity at the concentrations of 62.5 μg/mL and 125 μg/ mL, which dropped at higher concentrations. All propolis samples also showed antibacterial activity against the tested human pathogens similar to ampicillin and penicillin controls, except for Pseudomonas aeruginosa. However, they did not exert any antifungal activity against Candida albicans and Yarrowia lipolytica. In conclusion, propolis samples from both provinces showed promising biological activities, but further research should focus on finding the right concentrations for optimal effect and include the cell necrosis pathway to get a better idea of the anticarcinogenic effects.

          Abstract

          Cilj je ovoga istraživanja bio usporediti biološku aktivnost etanolnih ekstrakata propolisa pčelinje pasmine Apis mellifera caucasica iz dviju turskih provincija: Ardahana i Erzuruma. Testirana su njihova antioksidacijska, anticitotoksična, antikancerogena, antibakterijska i antifungalna svojstva. Uzorci iz tih dviju provincija razlikovali su se u mineralnom i organskom sastavu koji je odražavao njihovo zemljopisno podrijetlo. Test redukcije željeza/antioksidacijske snage (engl. ferric reducing antioxidant power, krat. FRAP) otkrio je superiornost ardahanskoga propolisa nad erzurumskim, no bez obzira na podrijetlo i prisutnost mitomicina Cu mediju, oba su propolisa povećala vijabilnost ljudskih perifernih limfocita, a učinak je ovisio o koncentraciji i trajanju. Propolis iz Erzuruma iskazao je najveću antikancerogenu aktivnost u koncentracijama od 62,5 i 125 μg/mL, no ona se smanjila s višim koncentracijama. Oba su propolisa također iskazala antibakterijsku aktivnost sličnu ampicilinskoj i penicilinskoj kontroli, osim kad se radilo o bakteriji Pseudomonas aeruginosa. Međutim, oba su zakazala protiv plijesni Candida albicans i Yarrowia lipolytica. Može se zaključiti da uzorci propolisa iz obiju provincija pružaju obećavajuće biološke aktivnosti, no u daljnja istraživanja, koja se trebaju usmjeriti na traženje optimalnih koncentracija za postizanje željenog učinka, treba uključiti i nekrotični put u mehanizmu djelovanja kako bi se stekao bolji uvid u njihovo antikancerogeno djelovanje.

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          The ferric reducing ability of plasma (FRAP) as a measure of "antioxidant power": the FRAP assay.

          A simple, automated test measuring the ferric reducing ability of plasma, the FRAP assay, is presented as a novel method for assessing "antioxidant power." Ferric to ferrous ion reduction at low pH causes a colored ferrous-tripyridyltriazine complex to form. FRAP values are obtained by comparing the absorbance change at 593 nm in test reaction mixtures with those containing ferrous ions in known concentration. Absorbance changes are linear over a wide concentration range with antioxidant mixtures, including plasma, and with solutions containing one antioxidant in purified form. There is no apparent interaction between antioxidants. Measured stoichiometric factors of Trolox, alpha-tocopherol, ascorbic acid, and uric acid are all 2.0; that of bilirubin is 4.0. Activity of albumin is very low. Within- and between-run CVs are <1.0 and <3.0%, respectively, at 100-1000 micromol/liter. FRAP values of fresh plasma of healthy Chinese adults: 612-1634 micromol/liter (mean, 1017; SD, 206; n = 141). The FRAP assay is inexpensive, reagents are simple to prepare, results are highly reproducible, and the procedure is straightforward and speedy. The FRAP assay offers a putative index of antioxidant, or reducing, potential of biological fluids within the technological reach of every laboratory and researcher interested in oxidative stress and its effects.
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            Antibacterial, antifungal and antiviral activity of propolis of different geographic origin.

            Propolis samples from different geographic origins were investigated for their antibacterial (against Staphylococcus aureus and Escherichia coli), antifungal (against Candida albicans) and antiviral (against Avian influenza virus) activities. All samples were active against the fungal and Gram-positive bacterial test strains, and most showed antiviral activity. The activities of all samples were similar in spite of the differences in their chemical composition. In samples from the temperate zone, flavonoids and esters of phenolic acids are known to be responsible for the above mentioned activities of bee glue; tropical samples did not contain such substances but showed similar activities. Obviously, in different samples, different substance combinations are essential for the biological activity of the bee glue. It seems that propolis has general pharmacological value as a natural mixture and not as a source of new powerful antimicrobial, antifungal and antiviral compounds.
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              Characterization of the cellular reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT): subcellular localization, substrate dependence, and involvement of mitochondrial electron transport in MTT reduction.

              The MTT assay, which is widely used to measure cell proliferation and to screen for anticancer drugs, is based on reduction of the tetrazolium salt, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) by actively growing cells to produce a blue formazan product. Despite broad acceptance of this assay, neither the subcellular localization, nor the biochemical events involved in MTT reduction are known. Mitochondrial involvement in MTT reduction has been inferred from studies with respiratory inhibitors using succinate as a substrate, but the contribution of this activity to overall cellular MTT reduction is unknown. Using the bone marrow-derived cell line, 32D, we investigated the subcellular localization of MTT reduction using succinate, NADH, and NADPH as substrates. At optimum substrate concentrations, MTT reduction by whole cell homogenates was greatest with NADH and least with succinate, which accounted for less than 10% of the combined activities. Using succinate, 96% of recoverable MTT reducing activity was in particulate fractions of the cell and 77% in the mitochondrial and light mitochondrial/lysosomal fractions. When NADH and NADPH were used as substrates, increased amounts of MTT reducing activity were associated with soluble fractions of the cell and association with mitochondrial fractions was less pronounced. To further characterize MTT reduction by the mitochondrial fraction, respiratory chain inhibitors were used to explore involvement of electron transport in MTT reduction. Succinate-dependent mitochondrial MTT reduction was inhibited by 80% with chlorpromazine, 70% by antimycin A, and 85-90% by thenoyltrifluoracetone (TTFA), but inhibition was not observed with rotenone at < or = 2 microM, Amytal, or azide. These results suggest that when succinate is used as an electron donor, 70-80% of mitochondrial MTT reduction occurs subsequent to transfer of electrons from cytochrome c to cytochrome oxidase, but prior to the point of azide inhibition. In contrast to succinate, NADPH-dependent mitochondrial MTT reduction was not affected by any of the respiratory inhibitors tested, and NADH-dependent reduction was only inhibited by chlorpromazine (40-50% at plateau concentrations). These results suggest that most cellular MTT reduction occurs outside the mitochondrial inner membrane and involves NADH and NADPH-dependent mechanisms that are insensitive to respiratory chain inhibitors. This interpretation is supported by whole cell studies in which rotenone failed to affect basal and interleukin-3-stimulated MTT reduction at times up to 4 h but strongly inhibited DNA synthesis. We conclude that most cellular reduction of MTT occurs extramitochondrially and probably involves the pyridine nucleotide cofactors NADH and NADPH.
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                Author and article information

                Journal
                Arh Hig Rada Toksikol
                Arh Hig Rada Toksikol
                aiht
                aiht
                Archives of Industrial Hygiene and Toxicology
                Sciendo
                0004-1254
                1848-6312
                March 2021
                30 March 2021
                : 72
                : 1
                : 53-69
                Affiliations
                [1 ]Ardahan University Faculty of Health Sciences, Department of Health Management , Ardahan, Turkey
                [2 ]Adıyaman University Faculty of Science and Letters, Department of Biology , Adıyaman, Turkey
                [3 ]Niğde Ömer Halisdemir University Faculty of Medicine, Department of Biophysics , Niğde, Turkey
                [4 ]Malatya Turgut Özal University, Battalgazi Vocational School , Battalgazi, Turkey
                [5 ]Ardahan University Faculty of Health Sciences, Department of Nursing , Ardahan, Turkey
                [6 ]Çukurova University Faculty of Ceyhan Engineering, Department of Chemical Engineering , Ceyhan, Turkey
                [7 ]Ardahan University, Ardahan Vocational School of Health Services , Ardahan, Turkey
                [8 ]Kahramanmaraş Sütçü İmam University Faculty of Science and Letters, Department of Biology , Kahramanmaraş, Turkey
                [9 ]Kilis 7 Aralık University Faculty of Science and Letters, Department of Molecular Biology and Genetics , Kilis, Turkey
                Author notes
                [* ] Yusuf Sevgiler, Adıyaman University Faculty of Science and Letters, Department of Biology, 02040 Adıyaman, Turkey ysevgiler@ 123456adiyaman.edu.tr
                Article
                aiht-2021-72-3492
                10.2478/aiht-2021-72-3492
                8191426
                33787188
                86c8312c-a2b0-4cd4-8ea0-309bbb40259a
                © 2021 Mehmet Arslan, Yusuf Sevgiler, Celal Güven, Zehra Tuğba Murathan, Nurcan Erbil, Deniz Yıldırım, Mehmet Büyükleyla, Şakire Karadaş, Rima Çelik, and Eyyüp Rencüzoğulları, published by Sciendo

                This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

                History
                : 01 September 2020
                : 01 September 2020
                : 01 March 2021
                Page count
                Pages: 17
                Categories
                Original Article

                anticarcinogen,antimicrobial,biological activity,mineral,mitomycin c,organic composition,antikancerogeno djelovanje,antimikrobno djelovanje,biološka aktivnost,minerali,mitomicin c,organski sastav

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