Assessment of Prevalence and Risk Factors for Diabetic Retinopathy in Patients with Type 1 and Type 2 Diabetes Examined at a Tertiary Care.

Introduction: Diabetic retinopathy (DR) is a microvascular complication of diabetes mellitus and the leading cause of visual impairment and blindness. The aim of the study was to estimate and compare the prevalence of DR and to determine an association between DR and systemic risk factors in hospitalized type 1 (DMT1) and type 2 (DMT2) diabetic patients. Material and methods: We analyzed 260 patients with diabetes, 43 with DMT1 and 217 with DMT2. The following data were collected: age, gender, type and duration of diabetes, glycemic control, blood pressure, estimated glomerular filtration rate, ophthalmologic examinations and routine biochemical parameters. Results: Out of the total number of 260 patients, 77 (29.6%) had non-proliferative DR (NPDR), 21 (8.1%) had proliferative DR (PDR), 29 (11.1%) had diabetic macular edema (DME), and 69 (23.5%) had diabetic cataracts. Forty-three (16.5%) patients were previously diagnosed with DMT1 and 217 (83.5%) with DMT2. The duration of diabetes was not significantly longer in DMT1 (12.8±11.2 years) in comparison to DMT2 (11.07±8.1 years). The prevalence of NPDR and PDR did not differ statistically in either groups. DME was more prevalent in DMT2 than in DMT1 (P<0.05). Diabetic cataract was found in 26.7% vs. 6.7% of patients with DMT2 and DMT1, respectively (p<0.01). The duration of diabetes significantly correlated with NPDR and PDR in DMT1 (r=o.31, p<0.05; r=0.55, p<0.001, respectively). In DMT2, significant correlations were found between the duration of diabetes and cataract, NPDR, PDR and DME (r=0.31, p<0.001; r=0.43 p<0.01, r=0.16 p<0.05 and r=0.20 p<0.01, respectively). Fasting plasma glucose (FPG) significantly correlated with PDR (r=0.258, p<0.05), while HbA1c with DME (r= 0.15 p<0.05). Conclusion: The duration of diabetes and hyperglycemia were associated with DR in both types of diabetes.

Gold(III) Complexes with Phenanthroline-derivatives Ligands Induce Apoptosis in Human Colorectal and Breast Cancer Cell Lines.

Due to their promising effects, gold(III) complexes recently drew increasing attention in the design of new metal-based anticancer therapeutics. Two gold(III) complexes, square-planar [Au(DPP)Cl2]+ - Complex 1 and distorted square-pyramidal [Au(DMP)Cl3] - Complex 2 (where DPP=4,7-diphenyl-1,10-phenanthroline and DMP=2,9-dimethyl-1,10-phenanthroline) were previously synthetized, described and approved as complexes with pronounced cytotoxic effects on colorectal HCT-116 and breast MDA-MB-231 cancer cells. This study investigated the type of cell death by AO/EB double staining, and identification of possible targets responsible for their cytotoxicity, monitored by immunofluorescence and qPCR methods. Both complexes induced apoptosis in all applied concentrations. In the HCT-116 cells apoptosis was activated by external apoptotic pathway, via increase of Fas receptor protein expression and Caspase 8 gene expression. Also, the mitochondrial pathway was triggered by affecting the Bcl-2 members of regulatory proteins and increased caspase 9 protein expression. In MDA-MB-231 cells, apoptosis was initiated from the mitochondria, due to disbalance between expressions of pro- and anti-apoptotic Bcl-2 family members and caspase 9 activation. Complex 1 shows better activity compared to Complex 2, which is in accordance with its structural characteristics. The results deal weighty data about proapoptotic activity of gold(III) complexes and highlighted potential targets for cancer therapy.

Royal Jelly and trans-10-Hydroxy-2-Decenoic Acid Inhibit Migration and Invasion of Colorectal Carcinoma Cells.

Research background: Acquisition of migratory potential is pivotal for cancer cells, enabling invasion and metastasis of colorectal carcinoma. Royal jelly and its bioactive component trans-10-hydroxy-2-decenoic acid (10H2DA) showed remarkable antimetastatic potential, but the molecular mechanism underlying this activity is unclear. Experimental approach: Identification and quantification of 10H2DA in royal jelly originating from Serbia was done by HPLC method. Cytotoxicity of 10H2DA was measured by tetrazolium dye MTT test in concentration range 1-500 µg/mL after 24 and 72 h. Its effect on the collective and single-cell migration was measured by wound healing and transwell migration assays. Invasive potential of cancer cells was evaluated by a transwell method modified with collagen. Immunofluorescence was used for migratory and invasive protein expression, while the gene expression of these markers was evaluated by quantitative real time polymerase chain reaction (qRT-PCR). All assays were applied on human colorectal carcinoma HCT-116 and SW-480 cell lines and, except for MTT, evaluated after 24 h of treatment with two selected concentrations of royal jelly and 10H2DA. Results and conclusions: According to HPLC, the mass fraction of 10H2DA in royal jelly was 0.92% (m/m). Treatment with 10H2DA showed no cytotoxic effect; however, significant inhibitory potential of royal jelly and 10H2DA on the motility and invasiveness of colorectal cancer cells was observed. More pronounced effect was exerted by 10H2DA, which significantly suppressed collective cell migration and invasiveness of SW-480 cells, as well as single- and collective cell migration and invasive potential of HCT-116 cell line. Treatments increased epithelial markers E-cadherin and cytoplasmic ß-catenin in HCT-116 cells, thus stabilizing intercellular connections. In SW-480 cells, 10H2DA increased E-cadherin on protein and gene level, and suppressed epithelial-mesenchymal transition (EMT) markers. In both cell lines, treatments induced significant suppression of promigratory/proinvasive markers: N-cadherin, vimentin and Snail on protein and gene level, which explains decreased migratory and invasive potential of HCT-116 and SW-480 cells. Novelty and scientific contribution: Our study presents new findings and elucidation of royal jelly and 10H2DA molecular mechanism that underlies their antimigratory/antiinvasive activity on colorectal cancer cells. These findings are shown for the first time indicating that these natural products are a valuable source of anticancer potential and should be reconsidered for further antitumour therapy.

Pseudevernia furfuracea inhibits migration and invasion of colorectal carcinoma cell lines.

ETHNOPHARMACOLOGICAL RELEVANCE: Pseudevernia furfuracea (L.) Zopf is common lichen species, traditionally used worldwide in treating various medical conditions, among which are intestinal issues and cancer. Most studies are focused mainly on cytotoxic potential of lichens, whilst their antimigratory and antiinvasive properties are often disregarded. Migration and invasion of cancer cells are pivotal processes in cancer metastasis, wherein cancer cells are able to migrate individually or in form of a coherent mass. One of successful strategies in anticancer treatments is targeting Wnt/ß-catenin signal pathway, that is aberrantly activated in colorectal carcinoma, as well as lowering level of migratory/invasive markers. AIM OF THE STUDY: Present study aimed to show antimigratory/invasive potential of Pseudevernia furfuracea methanol extract on HCT-116 and SW-480 colorectal carcinoma cell lines and to elucidate possible mechanism of its action. MATERIALS AND METHODS: Collective cell migration was assessed by Wound healing assay and single cell migration in real time by RTCA method. Analysis of anti- and promigratory protein expression was performed using immunofluorescent staining. Additionally, gene expression of antimigratory/promigratory and invasive (E-cadherin, ß-catenin, N-cadherin, Vimentin, Snail and MMP-9) markers were investigated by qRT-PCR method. Concentration of MMP-9 was determined colorimetrically by ELISA test. RESULTS: P. furfuracea extract was able to suppress both collective and single cancer cell migration, by inhibiting expression of promigratory/invasive markers and possibly re-establishing cell-cell adhesions. The present study indicates at P. furfuracea as effective antimigratory treatment, and HCT-116 cells were proved to be a more sensitive cell line to applied treatment. CONCLUSIONS: This lichen species is a promising candidate for application in treatment of cancer in order to prevent metastasis.

Modulation of cytotoxicity by consecutive adsorption of tannic acid and pesticides on surfactant functionalized zeolites.

This study investigated the environmental application of FAU type zeolites modified with cationic surfactants (cetylpyridinium chloride, tetrapropylammonium chloride and benzalkonium chloride). Adsorbent characterization was conducted using Fourier-transform infrared and Raman spectroscopy, thermogravimetry and differential thermal analysis, atomic force microscopy and X-ray powder diffraction. The efficiency for tannic acid adsorption from aqueous solution on the surface of prepared composites is studied and the adsorption process was modelled with different isotherm equations. Surfactant modifications of zeolites led to improved adsorption properties compared to FAU zeolites alone. The proposed mechanism controlling the adsorption of tannic acid onto surfactant modified zeolites mainly relies on π-π and hydrophobic interactions. The investigated materials are promising adsorbents for tannic acid and similar phenolics and may be important for environmental and dietary aspects of polyphenol persistence and usage. Further on, functionalized zeolites were studied for insecticide acetamiprid removal, prior to and after tannic acid retention. Promising findings of insecticide co-adsorption with tannic acid led to cytotoxicity evaluation. The cytotoxicity modulation effect of zeolites and tannic acid on acetamiprid points to the essential role of both components in insecticide toxicity reduction.

Hygrophorus eburneus, edible mushroom, a promising natural bioactive agent.

It is known that many edible mushrooms have important medicinal properties, including effects on different types of cancers. This is the first report regarding the neuroprotective, antimicrobial, antioxidative and anticancer activities of the acetone extract of edible mushroom Hygrophorus eburneus. Neuroprotective potential was evaluated by measuring the capacity of the extract to inhibit acetylcholinesterase. In this assay, the tested extract showed activity against acetylcholinesterase in a dose-dependent manner where the percentage of inhibition ranged from 13.19 to 46.44 %. The antimicrobial potential was determined by the microdilution method against five species of bacteria and eight species of fungi and the results of this method exhibited moderate antimicrobial activity of H. eburneus with MIC values ranging from 6.25 to 25 mg/mL. Antioxidant activity was evaluated by measuring the scavenging capacity of the tested sample on DPPH and superoxide anion radicals, by the reducing power assay and by measuring the amounts of total phenolics in extract. As a result of the study, H. eburneus extract showed a potent antioxidant activity (IC50 were 102.93 µg/mL for DPPH radical scavenging activity and 123.27 µg/mL for superoxide anion radicals scavenging) while absorbances for reducing power assay were from 0.0235 to 0.1161. The total phenolic content in the extract was 9.27 µg PE/mg. Finally, anticancer effects were evaluated by MTT test for cytotoxicity, acridine orange/ethidium bromide staining for detection of the type of cell death and wound healing assay for antimigratory effects on human colorectal cancer cell line (HCT-116) and human breast cancer cell line (MDA-MB-231). The results for cytotoxicity and apoptosis were measured after 24 and 72 h and for anti-migratory effect after 12 and 24 h. The tested H. eburneus mushroom extract expressed cell selectivity, with notable cytotoxic effects observed on HCT-116 cells, with a strong proapoptotic potential. The migration of HCT-116 cells was significantly inhibited, while MDA-MB-231 cells were less sensitive to the treatment. The results of this study revealed that the tested extract had relatively strong neuroprotective, antimicrobial, antioxidant, and anticancer effects. It suggests that this mushroom can be proposed as a novel source of nutraceuticals and pharmaceuticals.

The silk of Plodia interpunctella as a potential biomaterial and its cytotoxic effect on cancer cells.

Insect silk has been widely studied for its application in regenerative medicine. However, the data about Plodia interpunctella silk as a biomaterial and its anticancer properties are insufficient. Thus, the aim of this study was to investigate native silk as a substrate for growing normal human fibroblasts MRC-5, and test potential cytotoxic effects of the two silk extracts (with DMSO and Trypsin for sericin isolation) on HCT-116 colorectal carcinoma cells and MRC-5 fibroblasts as a control. Fifth-instar larval silk, collected for 15 and 30 days, was used for testing of proliferation and adhesion of MRC-5, 24 h and 72 h after seeding. Light- and fluorescence-microscope showed cell adhesion and spread on silk, as well as enhanced number of cells after 72 compared to 24 h and nonsignificant percentage of apoptotic cells on the silk. Although insoluble, P. interpunctella silk showed remarkable cytotoxic activity on HCT-116 cells, without significant cytotoxity on normal fibroblasts after 24 h and weak effects after 72 h. This study provides significant information about P. interpunctella silk as a potential biomaterial and shows the presence of some active constituents with anticancer properties, thus pointing to the possibility for exploitation of this worldwide pest insect in biomedical application.

Synthesis, characterization, DFT study, DNA/BSA-binding affinity, and cytotoxicity of some dinuclear and trinuclear gold(III) complexes.

In this study, we have synthesized a series of dinuclear and trinuclear gold(III) complexes of the general formula [Au2(N-N)Cl6] (1-3) for dinuclear and [Au3(N-N)2Cl8]+ (4-6) for trinuclear compounds, respectively, in which N-N is a bidentate ligand (1,4-diaminobutane; 1,6-diaminohexane or 1,8-diaminooctane). These complexes were characterized by elemental analysis, molar conductivity, and spectroscopic techniques (IR, UV-Vis, 1H NMR, ESI-MS). We performed DFT calculations to get insight into the geometry of the studies complexes. DNA-binding studies were performed by UV-Vis spectrophotometry and fluorescence spectroscopy. The results of competitive reactions between gold(III) complexes and ethidium bromide (EB) towards DNA have shown that selected complexes can displace EB from DNA-EB adduct. In addition, these experiments confirm that polynuclear gold(III) complexes interact with DNA covalently or via intercalation. Furthermore, high values of binding constants of gold(III) complexes towards bovine serum albumin (BSA) protein indicate good binding affinity. In addition, redox stability of complexes in the presence of DNA/BSA was confirmed by cyclic voltammetry. Results of the interactions between gold(III) complexes with DNA/BSA were discussed in reference to molecular docking data obtain by Molegro virtual docker. The cytotoxic activity of synthesized gold(III) complexes was evaluated on human breast cancer cell line (MDA-MB-231), human colorectal cancer cell line (HCT-116), and normal human lung fibroblast cell line (MRC-5). All complexes dose-dependently reduced cancer and normal cells viabilities, with significant cytotoxic effects (IC50 < 25 µM) for trinuclear gold(III) complexes (4, 5) on HCT-116 cells.

Potential of Teucrium chamaedrys L. to modulate apoptosis and biotransformation in colorectal carcinoma cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Teucrum chamaedrys L. is one of the known medicinal plants, useful for treatment of various health problems, especially digestive. In this study, we investigated methanol, ethyl-acetate and acetone extracts of T. chamaedrys in respect to their anticancer properties in SW480 colorectal cancer cells. MATERIALS AND METHODS: Cytotoxicity and proapoptotic potential were assessed by MTT cell viability assay and AO/EB double staining. Molecular mechanisms of induced apoptosis were determined by monitoring Fas receptor protein expression through immunofluorescence, Caspase 8 and 9 activity, as well as concentrations of O2.- spectrophotometrically. Additionally, mRNA expression of biotransformation enzymes (CYP1A1, CYP1B1, GSTP1) and membrane transporters (MRP1 and MRP2) involved in drug resistance were investigated by qPCR method. Qualitative analysis of individual phenolic compounds was performed by reversed phase HPLC-MS analysis. RESULTS: Methanol extract shows the best cytotoxicity and selectivity compared to ethyl-acetate and acetone extracts, mainly causing apoptosis of SW480 cells, without affecting normal HaCaT keratinocytes. The increased expression of Fas receptor protein and caspase 8 activity indicate that the death receptor-mediated pathway plays a crucial role in the observed apoptosis. The increased caspase 9 activity and O2.- concentration suggest that mitochondria are also involved in the apoptosis. T. chamaedrys methanol extract inhibits mRNA expression of CYP1A1, CYP1B1, GSTP1, MRP1 and MRP2 in SW480 cells. CONCLUSIONS: Induction of apoptosis and inhibition of CYP1A1, CYP1B1, GSTP1, MRP1 and MRP2 mRNA expression implies that T. chamaedrys can serve as a valuable source of bioactive compounds as dietary supplements or selective anticancer agents, with the ability to induce apoptosis and modulate drug resistance in colorectal cancer cells.

Proapoptotic and Antimigratory Effects of Pseudevernia furfuracea and Platismatia glauca on Colon Cancer Cell Lines.

The aim of this study is to investigate cytotoxic, proapoptotic, antimigratory and pro-antioxidant effects of methanol, acetone and ethyl acetate extracts of lichens Pseudevernia furfuracea and Platismatia glauca on colorectal cancer (HCT-116 and SW-480) cell lines. We compared the cytotoxic effects on colorectal cancer cells with the effects obtained from normal human fibroblast (MRC-5) cell line. Tetrazolium (MTT) test evaluated the cytotoxic effects, Transwell assay evaluated cell migration, acridine orange/ethidium bromide (AO/EB) fluorescent method followed the apoptosis, while prooxidant/antioxidant effects were determined spectrophotometrically through concentration of redox parameters. The tested extracts showed considerable cytotoxic effect on cancer cells with no observable cytotoxic effect on normal cells. Ethyl acetate and acetone extract of P. furfuracea induced the highest cytotoxicity (IC50=(21.2±1.3) µg/mL on HCT-116, and IC50=(51.3±0.8) µg/mL on SW-480 cells, respectively, after 72 h), with noteworthy apoptotic and prooxidant effects, and antimigratory potential of methanol extract. P. glauca extracts induced cytotoxic effects on HCT-116 cells after 72 h (IC50<40 µg/mL), while only methanol and acetone extracts had cytotoxic effects on SW-480 cells after 24 h, with proapoptotic/necrotic activity, as a consequence of induced oxidative stress. In conclusion, lichen extracts changed to a great extent cell viability and migratory potential of colorectal cancer cell lines. HCT-116 cells were more sensitive to treatments, P. furfuracea had better proapoptotic and antimigratory effects, and both investigated lichen species might be a source of substances with anticancer activity.

Naphthoquinone rich Onosma visianii Clem (Boraginaceae) root extracts induce apoptosis and cell cycle arrest in HCT-116 and MDA-MB-231 cancer cell lines.

In the present study, five root extracts of Onosma visianii Clem were investigated for their in vitro cytotoxic activity. On the basis of HPLC-PDA analysis, these extracts have proved to be a rich source of naphthoquinones as natural colourants for food and cosmetic industry. All investigated root extracts contain acetylshikonin, isobutyrylshikonin and α-methylbutyrylshikonin as major compounds. As the most abundant source of active compounds for antitumour therapy, acetone, chloroform and ethyl acetate extracts showed strong cytotoxic activity towards HCT-116 and MDA-MB-231 cancer cell lines. Also, these extracts induced apoptosis and cell cycle arrest in HCT-116 and MDA-MB-231 cancer cell lines.

Estradiol decreases blood pressure in association with redox regulation in preeclampsia.

In this study, we tested a hypothesis that a short-term estradiol therapy may reduce blood pressure in preeclampsia by modulating plasma oxidative stress. The intramuscular injections of 10 mg 17-beta-estradiol were prescribed to preeclamptic pregnant women during the 3-day therapy before a labor induction. The analyses of mean arterial pressure (MAP), serum estradiol concentrations, plasma superoxide anion (O2.), hydrogen peroxide (H2O2), nitrites (NO2-), and peroxynitrite (ONOO-) were conducted before and during the therapy. We found that the plasma concentrations of oxidative stress markers, such as O2- and H2O2, are higher in preeclampsia and positively correlated with the MAP value. Moreover, it was shown that the plasma concentration of NO2- as an indicator of NO levels is higher in preeclampsia. A short-term intramuscular application of estradiol decreases the MAP value and the plasma concentration of O.-, H2O2, NO2-, and ONOO- in preeclampsia. A positive correlation between the decrease of MAP values and the decrease of plasma concentrations of O2-, H2O2, and ONOO- was found in preeclampsia during a short-term estradiol therapy. We conclude that the short-term estradiol therapy decreases the MAP value in preeclampsia by modulating the plasma oxidative stress. We speculate that the estradiol metabolism in preeclampsia is an important mechanism that contributes to vascular dysfunction.

Novel seleno-hydantoin palladium(II) complex - antimigratory, cytotoxic and prooxidative potential on human colon HCT-116 and breast MDA-MB-231 cancer cells.

Selenium and palladium containing compounds separately exert multifunctional effects on cells. While selenium containing compounds usually exert antioxidative properties, palladium(II) containing compounds are cytotoxic and prooxidative. Here we investigated biological effects of bicyclic seleno-hydantoin cis-7a-ethyl-5-methyl-5-phenylselanylmethyl-tetrahydro-pyrrolo[1,2-c]imidazole-1,3-dione (Hid-Se), and its palladium(II) complex, trans-bis-(cis-7a-ethyl-5-methyl-5-phenylselanylmethyl-tetrahydro-pyrrolo[1,2-c]imidazole-1,3-dionato) palladium(II) chloride ((Hid-Se)2Pd) on human colon HCT-116 and breast MDA-MB-231 cancer cell lines. Hid-Se and (Hid-Se)2Pd showed prooxidative and cytotoxic character. In all performed experiments (Hid-Se)2Pd proved to be more active, i.e. this substance exerted greater prooxidative effect, cytotoxicity and influence on cell migration potential. Even though Hid-Se and (Hid-Se)2Pd enhanced migration of HCT-116 cells, very important feature of these substances is the strong antimigratory potential on metastatic MDA-MB-231 cells.

Quality of Life in Patients of Different Age Groups before and after Coronary Artery By-Pass Surgery.

PURPOSE: The study evaluates the changes in quality of life (QOL) six months after coronary artery bypass grafting (CABG) related to the patients' age. METHODS: The total of 243 consecutive patients completed the Nottingham Health Profile Questionnaire part 1 before and six months after CABG. Postoperative questionnaire was completed by 226 patients. Patients were divided into four examined groups (<50, 50-59, 60-69 and ≥70 years), according to their age. RESULTS: Six months after CABG, the quality of life in different sections has been significantly improved in most patients.The analysis of the relation between the age and the changes in QOL of patients six months after CABG showed a significant correlation among the patients' age and the improvement of QOL in the sections of physical mobility (r = 0.18, p = 0.008), social isolation (r = 0.17, p = 0.01) and energy ( r = 0.21, p = 0.002). The most prominent improvement was found in older patients. The age was not an independent predictor of QOL deterioration after CABG. CONCLUSIONS: The most noticeable improvement of QOL six months after CABG was found in older patients. Age is not the independent predictor of deterioration of QOL after CABG.

Antioxidative and antiproliferative evaluation of 2-(phenylselenomethyl)tetrahydrofuran and 2-(phenylselenomethyl)tetrahydropyran.

PURPOSE: To determine the antioxidant and antiproliferative influence of 2-(phenylselenomethyl)tetrahydrofuran (1a) and 2-(phenylselenomethyl)tetrahydropyran (2a) on colon cancer cell line HCT-116 and breast cancer cell line MDA-MB-231. METHODS: Cell viability was monitored in a dose-dependent manner using MTT assay. The concentration of superoxide anion radical (O2 •(-)) was determined spectrophotometrically. Spectrophotometric determination of nitrites (NO2 -) was performed by using the Griess method. Determination of total glutathione (GSH) was also performed spectrophotometrically. RESULTS: HCT-116 cell line was more sensitive to the effects of the investigated substances than MDA-MB-231 cell line. Also, it was noticed that 1a produced greater effect compared to 2a. Moreover, both investigated compounds decreased to a certain degree the oxidative stress by decreasing the O2•(-) and thus the peroxynitrite concentration. At the same time, 1a and 2a acted more efficiently in promoting the endogenous antioxidative capacities (increased GSH concentration) providing better self-defence capabilities for cells. CONCLUSION: Our findings showed that the investigated selenium compounds play an important role in reducing the levels of reactive oxygen species (ROS); therefore, we believe that, as antioxidants, they could prevent the processes arising as a consequence of oxidative stress, including cancer.

Radioprotectors - the evergreen topic.

To protect organisms from ionizing radiation (IR), and to reduce morbidity or mortality, various agents, called radioprotectors, have been utilized. Because radiation-induced cellular damage is attributed primarily to the harmful effects of free radicals, molecules with radical-scavenging properties are particularly promising as radioprotectors. Early development of such agents focused on thiol synthetic compounds, known as WR protectors, but only amifostine (WR-2721) has been used in clinical trials as an officially approved radioprotector. Besides thiol compounds, various compounds with different chemical structure were investigated, but an ideal radioprotector has not been found yet. Plants and natural products have been evaluated as promising sources of radioprotectors because of their low toxicity, although they exhibit an inferior protection level compared to synthetic thiol compounds. Active plant constituents seem to exert the radioprotection through antioxidant and free radical-scavenging activities. Our research established that plants containing polyphenolic compounds (raspberry, blueberry, strawberry, grape, etc.) exhibit antioxidative activities and protect genetic material from IR.

Chemical composition, cytotoxic and antioxidative activities of ethanolic extracts of propolis on HCT-116 cell line.

BACKGROUND: Propolis is a complex resinous sticky substance that honeybees collect from buds and exudates of various plants. Owing to its versatile biological and pharmacological activities, propolis is widely used in medicines, cosmetics and foods. The aim of this study was to evaluate the cytotoxic and antioxidative effects of various ethanolic extracts of propolis (EEPs) on human colon cancer cell line HCT-116 and compare them with their composition determined by HPLC-DAD. RESULTS: The most abundant flavonoids in all samples were chrysin, pinocembrin and galangin (12.697-40.811 µg mg⁻¹), while the main phenolic acids were caffeic acid, ferulic acid and isoferulic acid. Dose- and time-dependent inhibition of growth of HCT-116 cells was observed for all propolis samples, with IC50 values ranging from 26.33 to 143.09 µg mL⁻¹. Differences in cytotoxic activity of propolis samples were associated with differences in their composition. All EEP samples reduced both superoxide anion radical and nitrite levels and also had strong DPPH-scavenging activity. CONCLUSION: All tested propolis samples had pronounced cytotoxic and antioxidative activities.

Biological effects, total phenolic content and flavonoid concentrations of fragrant yellow onion (Allium flavum L.).

The antioxidant, antibacterial and antiproliferative activities, total phenolic content and concentrations of flavonoids of A. flavum extracts were determined. The total phenolic content was determined with Folin-Ciocalteu reagent and it ranged between 42.29 to 80.92 mg GA/g. The concentration of flavonoids in various extracts of A. flavum was determined using spectrophotometric method with aluminum chloride and obtained results varied from 64.07 to 95.71 mg RU/g. The antioxidant activity was monitored spectrophotometrically and expressed in terms of IC50 (µg/ml), and its values ranged from 64.34 to 243.34 µg/ml. The highest phenolic content and capacity to neutralize DPPH radicals were found in acetone extract. Antibacterial efficacy was defined by determining minimum inhibitory and minimum bactericidal concentrations using microdilution method. Significant antibacterial activity, especially for ethyl acetate extract, was observed. The best activity was showed against G+ bacteria, Staphylococcus aureus ATCC 25923 and Bacillus subtilis, while Escherichia coli was one of the least sensitive bacteria. Antiproliferative activity of the methanolic extract on HCT- 116 cell line was determined by MTT assay. Results showed that A. flavum has good antiproliferative activity with IC50 values of 28.29 for 24 h and 35.09 for 72 h. Based on these results, A. flavum is a potential source of phenols as natural antioxidant, antibacterial and anticancer substance of high value. Phenolic content of extracts depend on the solvents used for extraction.

Antiproliferative and proapoptotic activities of methanolic extracts from Ligustrum vulgare L. as an individual treatment and in combination with palladium complex.

The aim of this study is to examine the growth inhibitory effects of methanolic leaf and fruit extracts of L. vulgare on HCT-116 cells over different time periods and their synergistic effect with a Pd(apox) complex. The antiproliferative activity of plant extracts alone or in combination with the Pd(apox) complex was determined using MTT cell viability assay, where the IC(50) value was used as a parameter of cytotoxicity. Results show that antiproliferative effects of L. vulgare extracts increase with extension of exposure time, with decreasing IC(50) values, except for 72 h where the IC(50) values for methanolic leaf extract were lower than for the fruit extract. The Pd(apox) complex alone had a weak antiproliferative effect, but combination with L. vulgare extracts caused stronger effects with lower IC(50) values than with L. vulgare extracts alone. The type of cell death was explored by fluorescence microscopy using the acridin orange/ethidium bromide method. Treatments with plant extracts caused typical apoptotic morphological changes in HCT-116 cells and co-treatments with Pd(apox) complex caused higher levels of apoptotic cells than treatment with plant extracts alone. The results indicate that L. vulgare is a considerable source of natural bioactive substances with antiproliferative activity on HCT-116 cells and which have a substantial synergistic effect with the Pd(apox) complex.

Antioxidant, antimicrobial and antiproliferative activities of five lichen species.

The antioxidative, antimicrobial and antiproliferative potentials of the methanol extracts of the lichen species Parmelia sulcata, Flavoparmelia caperata, Evernia prunastri, Hypogymnia physodes and Cladonia foliacea were evaluated. The total phenolic content of the tested extracts varied from 78.12 to 141.59 mg of gallic acid equivalent (GA)/g of extract and the total flavonoid content from 20.14 to 44.43 mg of rutin equivalent (Ru)/g of extract. The antioxidant capacities of the lichen extracts were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging. Hypogymnia physodes with the highest phenolic content showed the strongest DPPH radical scavenging effect. Further, the antimicrobial potential of the lichen extracts was determined by a microdilution method on 29 microorganisms, including 15 strains of bacteria, 10 species of filamentous fungi and 4 yeast species. A high antimicrobial activity of all the tested extracts was observed with more potent inhibitory effects on the growth of Gram (+) bacteria. The highest antimicrobial activity among lichens was demonstrated by Hypogymnia physodes and Cladonia foliacea. Finally, the antiproliferative activity of the lichen extracts was explored on the colon cancer adenocarcinoma cell line HCT-116 by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) viability assay and acridine orange/ethidium bromide staining. The methanol extracts of Hypogymnia physodes and Cladonia foliacea showed a better cytotoxic activity than the other extracts. All lichen species showed the ability to induce apoptosis of HCT-116 cells.