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Dunaliella bardawil is a carotenoid-producing alga that is being considered for use in nutraceuticals. To evaluate potential protective effects of consumption of this alga, rats were treated with two different doses of D. bardawil (2.5 and 5.0 g kg-1 body weight [bw]) as a biomass suspension daily for 14 days. Animals were tested against Carbon tetrachloride (CCl4; 2 ml kg-1)–induced liver tox-icity as measured by various biochemical marker enzymes in liver and blood. All measurements were taken 6 h following the single dose of CCl4. The results of this study show that there was a slight, but statistically significant mean serum enzyme values, with D. bar-dawil treatment, compared to higher mean values in animals receiving CCl4 alone. Lipid peroxidation is measured by thiobarbituric acid–reactive substance (TBARS) activity was likewise slightly less elevated with algae treatment. The results also demonstrated protection against DNA strand breaks in hepatocytes, as measured by single cell gel electrophoresis. Liver histopathology was less severe with D. bardawil treatment, supporting the apparent protective action of 14-day treatment on hepatic oxidative injury. [ABSTRACT FROM PUBLISHER]

The current protocol of cancer management includes surgery, radiotherapy and chemotherapy. However, these modalities have significant adverse effects and affect the quality of life. Further intensification of treatment is hindered as maximal toxicity levels are reached impeding improvement. Hence researchers are in the quest for adjunctive naturally available therapies that can alter tumor proliferation without causing significant adverse reactions. The present study aims to explore the cytotoxic potential of earthworm coelomic fluid (ECF) of Eudrilus eugeniae (EE), Eisenia foetida (EF), and Perionyx excavatus (PE) on oral cancer cell line SCC-9. The effect of ECF on cell cycle analysis and mechanism of cell death have also been investigated. All experiments reported in this paper were performed as 3 replicates per experiment. The results indicated that ECF of EE, EF and PE have potent variable cytotoxic effect on SCC-9 cells demonstrated through LDH, clonogenic and comet assay. An effective cell cycle arrest was observed at the G2M phase of cell cycle with apoptotic induction that was observed through an Annexin V - FITC/PI assay. ECF of EE was found to be superior in its cytotoxic action closely followed by ECF of PE. The present findings provide evidence for the first time that ECF of EE, EF and PE have potent cytotoxic effect on oral cancer cells in vitro . They significantly induce G2M cell cycle arrest and promote apoptosis in SCC-9 cell line. Gene expression studies have been planned to ascertain the pathways of cell death.

Introduction: The earthworm coelomic fluid (ECF) has shown proven antiproliferative effect against breast, liver, gastrointestinal, and brain cancer, but it is least explored in oral cancer. The present in vitro study is an attempt to investigate the antiproliferative activity of ECF on oral cancer cell line squamous cell carcinoma (SCC)-9., Materials and Methods: ECF was collected from the species Eudrilus eugeniae (EE), Eisenia foetida (EF), and Perionyx excavatus (PE) stored at -80°C. Percentage inhibition of ECF on squamous cell carcinoma-9 cells in vitro was recorded at 24 h. Protein estimation was done using Bradford protein assay validated by the biuret method. Cytotoxicity was tested at 2.5, 5, 10, 20, 40, and 80 μg/ml concentrations by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay in SCC-9 cells in vitro . GraphPad Prism 7.0 software was used to calculate the inhibitory concentration (IC 50 ). Chi-square test was used to analyze the difference between samples., Results: The test samples EE, EF, and PE inhibited the growth of SCC-9 cells significantly in a dose-dependent manner, and the IC 50 values were found to be 4.6, 44.69, and 5.27 μg/ml, respectively. The antiproliferative effect was found to be variable among the three earthworm species with EE showing the most promising effect followed by PE and EF., Conclusion: Establishing the antiproliferative effect of ECF on oral cancer cells could be an initial step toward drug development and future anticancer research. The preliminary investigation has shown that ECF has a promising antiproliferative effect on oral cancer cells in vitro ., Summary: The present pilot study evaluated the in vitro antiproliferative effect of earthworm coelomic fluid (ECF) of Eudrilus eugeniae (EE), Eisenia foetida (EF), and Perionyx excavatus (PE) on squamous cell carcinoma-9 cell line. The ECF inhibitory activity was promising at inhibitory concentration values of 4.6, 44.69, and 5.27 μg/ml, respectively. Further studies pertaining to antiproliferative mechanism of EE, EF, and PE have been planned. Abbreviations Used: ECF: Earthworm coelomic fluid, EE: Eudrilus eugeniae , EF: Eisenia foetida , PE: Perionyx excavatus , MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, SCC: Squamous cell carcinoma, BSA: Bovine serum albumin, PBS: Phosphored buffered saline, ATCC: American Type Culture Collection., Competing Interests: There are no conflicts of interest.

A number of studies in the recent years have evaluated the anti-proliferative activity of flavonoids. Although certain studies investigated the structure-activity based on the phenotypic assays, no study has correlated the flavonoids structure with the ability to alter gene/protein expression. Present study was focused to understand the structure-function relationship of citrus flavonoids in terms of their ability to alter the gene expression in the colon adenocarcinoma cells. Eight structurally related flavonoids found in citrus were evaluated for their ability to inhibit colon cancer (SW480) cells, as well as change the expression of apoptosis related genes/proteins. Apigenin and quercetagetin demonstrated most significant inhibition of cell proliferation with 63.6% and 45.7% inhibition of cell growth at 200μM after 48h of incubation, respectively. The cell death was also confirmed by images of fluorescently tagged cells. Furthermore, up-regulation of Bax/Bcl2 protein ratio as well as activation of Caspase3 at 200μM at 48h confirmed the induction of apoptosis by apigenin and quercetagetin. In addition, results suggest that the change in Bax/Bcl2 ratio by apigenin and quercetagetin seems to be due to their ability to alter the expression of bax and bcl2 transcription. Results of the currents study suggest that among the citrus flavonoids, double bond between C2 and C3 and hydroxyl group at C3, C6 are highly decisive for the proliferation inhibition and apoptosis induction ability. Taken together, these results demonstrate that among the major flavonoids of citrus, apigenin and quercetagetin have potent anti-cancer activity through inducing apoptosis in SW480 human colon cancer cells., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Objectives: The study was aimed to purify obacunone and obacunone glucoside (OG) from seeds of Marsh White grapefruit and understand the mode of cytotoxicity of limonoids on colon cancer (SW480) cells., Methodology: Both limonoids were purified using chromatographic techniques. The structures and purity of limonoids were confirmed by NMR and HPLC analysis, respectively., Results: Obacunone and OG inhibited SW480 cell proliferation with IC50 values of 97 and 109.7 μM respectively, at 24h. Sequence of events such as decreased ratio of bcl2/bax gene transcription, activation of caspase-3, fragmentation of DNA in cells treated with obacunone and OG demonstrated induction of apoptosis by limonoids. Additionally, higher induction of cytochrome-c in cytosol suggests the activation of intrinsic apoptosis by limonoids. Involvement of apoptosis was also confirmed through expression of bax, bcl2, pro-caspase-3 and caspase-9. Both the limonoids activated p21 and arrested cells at G1 and G2/M phase. Additive activity of proliferation inhibition and activation of caspase-3 by limonoids was observed when combined with camptothecin, demonstrating the induction of apoptosis. In conclusion, both limonoids induced apoptosis by activation of intrinsic apoptosis pathway and activation of p21 leading to arresting cells at G2/M phase of the cell cycle., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

This is the first report on the hepatoprotective potentials of marine micro algae Dunaliella species. Dunaliella salina, halotolarent green alga was cultivated in modified autotrophic medium. The alga was subjected to light and nutrient stress in order to accumulate (beta-carotene along with other carotenoids. Such beta-carotene enriched yellow cells were fed to rats by mixing with regular feed at the dose of 2.5 and of 5.0gkg(-1) b.w. for 2 weeks. The degree of hepatoprotection was measured up on challenging animals with toxin (2.0gkg(-1) of carbon tetrachloride) by estimation of biochemical parameters like, serum transaminases (serum aspartate transaminase (S)AST and serum alanine transaminase (S)ALT), serum alkaline phosphatase and total protein. The results were compared to animals on normal diet and with group fed with 100mugkg(-1) b.w. of standard all trans beta-carotene. Among the three test groups the group fed with algae of 5.0gkg(-1) body weight, showed maximum protection. The levels of (S)AST and (S)ALT was found to be 61.3+/-6.4 and 80.7+/-5.6%, against 90.8+/-10.5 and 144.7+/-13.9% in case of standard beta-carotene. The protein contents were increased in case of control to 6.1+/-0.7 and the same was found to be significantly less in case of 5.0gkg(-1)Dunaliella fed group, which shown 5.6+/-0.8% total protein. However, the activity of 2.5gkg(-1) was also significant comparatively (P<0.05). The results indicate that Dunaliella, which contains isomeric forms of beta-carotene can act as good antihepatotoxic when compared to synthetic all trans beta-carotene. Dunaliella has shown the presence of both cis and trans isomeric forms of beta-carotene, where as synthetic compounds contain only trans isomer. Hepatoprotectivity may be due to presence of various isomeric forms of carotene and other oxygenated carotenoids (xanthophylls) in algae.

Dunaliella salina a green marine alga is known for its carotenoid accumulation, having various applications in the health and nutritional products. The purpose of present study was to evaluate the ability of D. salina algal powder extract to protect against oxidative stress In vivo using animal models. Treatment of albino Wistar strain rats with 125 microg/kg and 250 microg/kg b.w. showed significant protection when compared to toxin treated (CCl4) group. Since beta-carotene is major constituent of Dunaliella the results were also compared with group treated with 250 microg/kg b.w (p.o.) synthetic all trans beta-carotene. Treatment of CCl4 at dose of 2.0 g/kg b.w. decreased the activities of various antioxidant enzymes like catalase, superoxide dismutase (SOD) and peroxidase by 45.9%, 56% and 54% respectively compared to control group and lipid peroxidation value increased nearly 2 folds. Pretreatment of rats with 125 microg carotenoid followed by CCl4 treatment caused restoration of catalase, SOD and peroxidase by 25.24%, 23.75 and 61.15% respectively as compared to control. The group treated with 250 microg/kg has shown the restoration of 53.5%, 57.7 and 90.64% of catalase, SOD and peroxidase, respectively. This group has shown 75.0% restoration of peroxidation compared to control group of animals. The above enzyme activities were not significantly restored in group treated with synthetic all trans beta-carotene, which showed 7.5%, 23.8% restore in catalase and peroxidase content. The level of superoxide dismutase remained same and lipid peroxidation value decreased only by 23% in synthetic all trans beta-carotene treated group in comparison with control group. These results clearly indicate the beneficial effect of algal carotenoid compared to synthetic carotene as antioxidant. Owing to this property, the algae Dunaliella can be further extended to exploit, its possible application for various health benefits as nutraceuticals and food additive.

Extracts of Cissus quadrangularis L. were tested for antioxidant activity by beta-carotene linoleic acid model and also by 1,1-diphenyl-2-picrylhydrazyl model. The ethyl acetate fraction of both fresh and dry stem extracts at a concentration of 100 ppm showed 64.8% antioxidant activity in the beta-carotene linoleic acid system and 61.6% in the 1,1-diphenyl-2-picrylhydrazyl system. This fraction showed the presence of sterols, vitamin C, and tannins as phytoconstituents. The antioxidant activity of methanol extract and aqueous extract were comparatively less significant than that of ethyl acetate extract, and n-hexane extract showed the least activity. The ethyl acetate extract and methanol extract of both fresh and dry stems further exhibited antimicrobial activity against Gram-positive bacteria, including Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, and Streptococcus species. The results of the study have implications in the use of C. quadrangularis as an antibacterial agent and more so as an antioxidant in several applications requiring these properties.

Antioxidant-rich fractions were extracted from pomegranate (Punica granatum) peels and seeds using ethyl acetate, methanol, and water. The extracts were screened for their potential as antioxidants using various in vitro models, such as beta-carotene-linoleate and 1,1-diphenyl-2-picryl hydrazyl (DPPH) model systems. The methanol extract of peels showed 83 and 81% antioxidant activity at 50 ppm using the beta-carotene-linoleate and DPPH model systems, respectively. Similarly, the methanol extract of seeds showed 22.6 and 23.2% antioxidant activity at 100 ppm using the beta-carotene-linoleate and DPPH model systems, respectively. As the methanol extract of pomegranate peel showed the highest antioxidant activity among all of the extracts, it was selected for testing of its effect on lipid peroxidation, hydroxyl radical scavenging activity, and human low-density lipoprotein (LDL) oxidation. The methanol extract showed 56, 58, and 93.7% inhibition using the thiobarbituric acid method, hydroxyl radical scavenging activity, and LDL oxidation, respectively, at 100 ppm. This is the first report on the antioxidant properties of the extracts from pomegranate peel and seeds. Owing to this property, the studies can be further extended to exploit them for their possible application for the preservation of food products as well as their use as health supplements and neutraceuticals.