1. 6. References 1 The Hebrew University of Jerusalem, Faculty of Medicine, School of Pharmacy, The Institute for Drug Research, Jerusalem, Israel 1. Background The aim of this research was to find out the proliferation inhibition properties in vitro of Chilean and Polish berries [Myrteola nummularia, ‘Murtilla-like’] vs. well known ‘Murtilla’, blueberries, and raspberries and the possible correlations with bioactive compounds and the antioxidant activities (AA). Dimethylsulfoxide (DMSO) extracts of berries on human colorectal cancer (CRC) cell lines HT ‑ 29 and SW48 proliferation were investigated. 2. Materials and methods Bioactive compounds and the antioxidant activities in berries were determined by radical scavenging assays. The interaction between bovine serum albumin (BSA) and quercetin, and BSA and berry extracts was measured by 3-dimensional fluorescence (3D-FL) and FTIR (1-4). MTT assay, flow cytometric analyses of cell cycle and apoptosis were used. 3. Results 4. Discussion Figure 1. A, Box/Whisker and Dot Plots of DPPH-free radical activity of DMSO extracts; B, Principal component analysis (PCA), based on DPPH measurements of DMSO extracts of MNR- ‘Murtilla’ non-ripe, MLN- ‘Murtilla-like’ non-ripe, BP- blueberry Poland, BC- blueberry Chile, and R- raspberry; Extract concentrations (1- 25 mgml -1 ) and DPPH reaction times (1 – 90 min) were used. Figure 4. Changes in the fluorescence intensity as a result of binding affinity (A): 2 x 10 -5 M/L BSA (first line from the top) and 8  g ‘Murtilla’ (second line from the top) with addition of 0.88 x 10 -5 M/L quercetin after 20 min (last line from the top). The reaction times: 0, 2, 5, 10, 15, and 20 min; (B): The same conditions as in (A), but quercetin and ‘Murtilla’ extracts were added immediately to BSA at 0 time (second line from the top); (C) contour map (from 3 D-FL spectra) illustrates the quenching results after reaction time of 20 min as in (A). DPPH (1, 1-diphenyl-2-picrylhydrazyl radical) kinetic measurements (Fig. 1A) showed that the quenching ability of the DMSO extracts were comparable between them, with the exception of higher values of ‘Murtilla’ non-ripe berries. The plot of principal components (Fig. 1B) illustrates the large variability of the investigated berries. PCA segregated from the rest berries clearly only one group of ‘Murtilla’ non-ripe with the highest antiradical activity. 5. Conclusions 1.Gorinstein, S., Leontowicz, H., Leontowicz, M., Jesion, I., Namiesnik, J., Drzewiecki, J., Park, Y.S, Ham, K.S., Giordani, E., & Trakhtenberg, S. (2011). Nutrition 1.Gorinstein, S., Leontowicz, H., Leontowicz, M., Jesion, I., Namiesnik, J., Drzewiecki, J., Park, Y.S, Ham, K.S., Giordani, E., & Trakhtenberg, S. (2011). Nutrition, 27, 838-846. 2.Park, Y.S., Leontowicz, H., Leontowicz, M., Namiesnik, J., Suhaj, M., Cvikrova, M., Martincova, O., Weisz, M., & Gorinstein, S. (2011), Journal of Food Composition and Analysis 2.Park, Y.S., Leontowicz, H., Leontowicz, M., Namiesnik, J., Suhaj, M., Cvikrova, M., Martincova, O., Weisz, M., & Gorinstein, S. (2011), Journal of Food Composition and Analysis, 24, 963-970. 3.Gorinstein, S., Poovarodom, S., Leontowicz, H., Leontowicz, M., Namiesnik, J., Vearasilp, S., Haruenkit, R., Ruamsuke, P., Katrich, E., & Tashma, Z. (2011). Food Research International 3.Gorinstein, S., Poovarodom, S., Leontowicz, H., Leontowicz, M., Namiesnik, J., Vearasilp, S., Haruenkit, R., Ruamsuke, P., Katrich, E., & Tashma, Z. (2011). Food Research International, 44, 2222-2232. European Journal of Integrative Medicine 4.Leontowicz, H., Leontowicz, M., Jesion, I., Bielecki, W., Poovarodom, S., Vearasilp, S., González- Aguilar, G., Robles-Sánchez, M., Trakhtenberg, S., & Gorinstein, S., 2011. European Journal of Integrative Medicine, 3, 169–181. INHIBITION OF CANCER CELL PROLIFERATION IN VITRO BY DIFFERENT BERRIES AND CORRELATION WITH THEIR ANTIOXIDANT ACTIVITIES Shela Gorinstein #1, Yong Seo Park 2,Sylwia Flis 3, Zenon Jastrzebski 3, Patricia Arancibia-Avila 4, Milan Suhaj 5, Elena Katrich 1, Moshe Weisz 1, Zeev Tashma 1 Shela Gorinstein #1, Yong Seo Park 2, Sylwia Flis 3, Zenon Jastrzebski 3, Patricia Arancibia-Avila 4, Milan Suhaj 5, Elena Katrich 1, Moshe Weisz 1, Zeev Tashma 1 Figure 2. A, Survival of human colorectal cancer (CRC) cells of the SW48 and HT ‑ 29 lines. B, Changes in the cell cycle progression of human CRC cells after 72 h of treatment with berries extracts: blueberry Poland (BBP), blueberry Chile (BBCh), 'Murtilla' (MT) and 'Murtilla-like' (MTL). Figure 3. Induction of cell death by berries extracts in the human CRC cells. Death cells (%) was determined by FACS analysis after 72 h of treatment. Staining - annexin V-FITC/PI. The induction of cell death was significant (*) in comparison with control (P<0.05). Cells stained with Calcein-AM and PI (10×). Green fluorescence indicates living cells, red fluorescence - dead cells. Blueberry Poland (BBP), blueberry Chile (BBCh), 'Murtilla' (MT) and 'Murtilla-like' (MTL). The berry extracts decreased the proliferation of both CRC cells (Figs. 2, 3), and the effect was concentration dependent. The inhibition effect of growth of human CRC cells for the highest concentration of the extracts varied 2-3- fold among the berries. The lowest IC 50 values 751 and 858 µg/ml were for MT extract on HT-29 and SW48. HT-29 cells treated with MT showed a decrease in G1 phase cells from 77% to 56%. Death cells (%) treated with MT were 80.1 and 72.5 for SW48 and HT-29 cells, respectively. Fluorescence emission system of BSA was studied with increased concentrations of flavonoids and berry extracts and with increased duration of reaction time (Fig. 4). After interaction with quercetin and berries extracts FTIR spectrum of BSA showed a shift in the maximum of Amides I and II in the region of 1650 and 1550 cm -1, respectively. There were differences in the contents of the analyzed antioxidants in the extracts: the highest amount of polyphenols, flavonoids and antioxidant activities were in 'Murtilla' berry extract (1-4). DPPH kinetic measurements were used to compare, distinguish and discriminate the antiradical activity among berry extracts by multivariate analysis. It is known that the interaction between drugs and serum albumin plays an important role in the distribution and metabolism of drugs, therefore the interaction between two kinds of flavonoids (catechin and quercetin) and BSA and berries extracts was investigated by 3D- FL and FTIR spectroscopy (4). The results of this study indicated that flavonoids have strong ability to quench the intrinsic fluorescence of BSA by forming complexes. The interaction between polyphenol extracts of 'Murtilla- like' and BSA showed that the new kind of berries has a strong ability as other studied samples to decrease the intrinsic fluorescence of BSA. These findings suggest that the intake of a new kind of berry, as a source of natural antioxidants, may reduce colon cancer risk. # This research was done in memory of my dear brother Prof. Simon Trakhtenberg, who died in November 2011, who encouraged and supported me during all his life A B A B AB C 3 Department of Pharmacology, National Medicines Institute, Warsaw, Poland 4 Department of Basic Sciences, Universidad del Bio-Bio, Chillan, Chile 5 Food Research Institute, 824 75 Bratislava, Slovakia Our publications- http://www.bashanfoundation.org/shela/shelapub.htmlhttp://www.bashanfoundation.org/shela/shelapub.html The inhibition of cancer cell proliferation correlated with the levels of polyphenols, flavonoids and their antioxidant activities. 2 Department of Horticultural Science, Mokpo National University, Muan, South Korea