Garcinol, or polyisoprenylated benzophenone, isolated from the rind of fruiting bodies of Garcinia indica, has been used in traditional medicine for its potential antiinflammatory and antioxidant properties. The objective of this study was to investigate the effect of garcinol on pancreatic cancer (PaCa) cell viability and proliferation. For this, 2 human PaCa cell lines, BxPC-3 and Panc-1, with wild and mutant k-ras, respectively, were treated with garcinol (0-40 μM). Garcinol significantly (P < 0.05) inhibited cell growth (trypan blue exclusion) by induction of apoptosis in a dose- and time-dependent manner. Flow cytometric analysis revealed G0-G1 phase cell cycle arrest in both cell lines. The molecular mechanism of garcinol's action on PaCa cells was investigated by targeting signaling moieties involved in apoptosis (X-IAP, cIAP, caspase-3, 9, and PARP cleavage), transcription factor NF-κB, believed to contribute toward a chemoresistance phenotype in pancreatic tumors, and molecules associated with neovascularization and metastasis (MMP-9, VEGF, IL-8, and PGE(2)). Garcinol significantly (P < 0.05) augmented antiproliferative, proapoptotic, antimetastatic, and antiangiogenic effects in both PaCa cell types relative to untreated cells. These effects were more pronounced in Panc-1. This is the first report on the therapeutically relevant effect of garcinol in PaCa. Further studies are warranted, based on our findings.

Advanced glycation endproducts (AGE) are oxidative products formed from the reaction between carbohydrates and a free amino group of proteins that are provoked by reactive species (RS). It is also known that AGE enhance the generation of RS and that the binding of AGE to a specific AGE receptor (RAGE) induces the activation of the redox-sensitive, pro-inflammatory transcription factor, nuclear factor-kappa B (NF-kB). In this current study, we investigated the anti-oxidative effects of short-term kaempferol supplementation on the age-related formation of AGE and the binding activity of RAGE in aged rat kidney. We further investigated the suppressive action of kaempferol against AGE's ability to stimulate activation of pro-inflammatory NF-kB and its molecular mechanisms. For this study, we utilized young (6 months old), old (24 months old), and kaempferol-fed (2 and 4 mg/kg/day for 10 days) old rats. In addition, for the molecular work, the rat endothelial cell line, YPEN-1 was used. The results show that AGE and RAGE were increased during aging and that these increases were blunted by kaempferol. In addition, dietary kaempferol reduced age-related increases in NF-kappaB activity and NF-kB-dependant pro-inflammatory gene activity. The most significant new finding from this study is that kaempferol supplementation prevented age-related NF-kappaB activation by suppressing AGE-induced nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase). Taken together, our results demonstrated that dietary kaempferol exerts its anti-oxidative and anti-inflammatory actions by modulating the age-related NF-kappaB signaling cascade and its pro-inflammatory genes by suppressing AGE-induced NADPH oxidase activation. Based on these data, dietary kaempferol is proposed as a possible anti-AGE agent that may have the potential for use in anti-inflammation therapies.

Natural antioxidants such as polysaccharides with strong antioxidant activities are used to protect against oxidative damage, yet little is known so far about the antioxidant effects of litchi fruit polysaccharides. In the present study, four different polysaccharide-enriched fractions were isolated from litchi pulp tissue and partially purified by a stepwise method of ethyl alcohol (EtOH) precipitation. Their chemical and physical characteristics were determined by chemical methods, gas chromatography and IR spectrophotometry. Antioxidant activities of these fractions were investigated using various in vitro assay systems. These four polysaccharide-enriched fractions exhibited a dose-dependent free radical scavenging activity as shown by their DPPH radical, superoxide anion and hydroxyl radical inhibition, chelating ability and reducing power. Among the different fractions, LFP-III showed the strongest scavenging activity against DPPH radical, superoxide and hydroxyl radicals and chelating ability. These findings suggest litchi polysaccharides from pulp tissue have potential as functional foods with enhanced antioxidant activity.

Whether garcinol, the active component of Garcinia indica, can modulate the sensitivity of cancer cells to TRAIL, a cytokine currently in phase II clinical trial, was investigated. We found that garcinol potentiated TRAIL-induced apoptosis of cancer cells as indicated by intracellular esterase activity, DNA strand breaks, accumulation of the membrane phospholipid phosphatidylserine, mitochondrial activity, and activation of caspase-8,-9, and -3. We found that garcinol, independent of the cell type, induced both of the TRAIL receptors, death receptor 4 (DR4) and DR5. Garcinol neither induced the receptors on normal cells nor sensitized them to TRAIL. Deletion of DR5 or DR4 by small interfering RNA significantly reduced the apoptosis induced by TRAIL and garcinol. In addition, garcinol downregulated various cell survival proteins including survivin, bcl-2, XIAP, and cFLIP, and induced bid cleavage, bax, and cytochrome c release. Induction of death receptors by garcinol was found to be independent of modulation of CCAAT/enhancer-binding protein-homologous protein, p53, bax, extracellular signal-regulated kinase, or c-Jun-NH(2)-kinase. The effect of garcinol was mediated through the generation of reactive oxygen species, in as much as induction of both death receptors, modulation of antiapoptotic and proapoptotic proteins, and potentiation of TRAIL-induced apoptosis were abolished by N-acetyl cysteine and glutathione. Interestingly, garcinol also converted TRAIL-resistant cells into TRAIL-sensitive cells. Overall, our results indicate that garcinol can potentiate TRAIL-induced apoptosis through upregulation of death receptors and downregulation of antiapoptotic proteins. Mol Cancer Ther; 9(4); 856-68.(c)2010 AACR.

Previous studies have demonstrated that the persistent exposure of human bronchial epithelial cells to nicotine (Nic) through nicotinic acetylcholine receptors increases cyclin D1 promoter activity and protein expression. The main purpose of this study is to elucidate the carcinogenic role of cyclin D3, which is involved in breast tumorigenesis when induced by Nic. Real-time PCR analysis revealed that cyclin D3 is highly expressed at the mRNA level in surgically dissected breast tumor tissue, compared to the surrounding normal tissue (tumor/normal fold ratio = 17.93, n = 74). To test whether Nic/nicotinic acetylcholine receptor (nAChR) binding could affect cyclin D3 expression in human breast cancer cells, the transformed cell line MCF-10A-Nic (DOX) was generated from normal breast epithelial cells (MCF-10A) with inducible α9-nAChR gene expression, using the adenovirus tetracycline-regulated Tet-off system. Tet-regulated overexpression of α9-nAChR in MCF-10A-Nic (DOX) xenografted BALB/c-nu/nu mice resulted in a significant induction of cyclin D3. In contrast, cyclin D3 expression was down-regulated in α9-nAChR knock-down (siRNA) MDA-MB-231-xenografted tumors in NOD.CB17-PRKDC(SCID)/J(NOD-SCID) mice. Furthermore, we found that Nic-induced human breast cancer (MDA-MB-231) cell proliferation was inhibited by 1 μM of garcinol (Gar), isolated from the edible fruit Garcinia indica, through down-regulation of α9-nAChR and cyclin D3 expression. These results suggest that α9-nAChR-mediated cyclin D3 overexpression is important for nicotine-induced transformation of normal human breast epithelial cells. The homeostatic regulation of cyclin D3 has the potential to be a molecular target for antitumor chemotherapeutic or chemopreventive purposes in clinical breast cancer patients.

Using albumin as model, we conducted series of in vitro glycation experiments to examine role of zinc in glycation using glucose at 4-100 mg/ml, incubations at 37°C or 60°C, duration of 2 or 4 weeks and in presence of zinc or ascorbic acid (AA) or folic acid (FA). Modifications of bovine serum albumin (BSA) were examined by using fluorescence of advanced glycation end products (AGEs) and dityrosine, UV, and Fourier transformed infrared spectroscopy. Adding zinc (0 to 768.5 μmol/l) resulted in significant inhibition of albumin glycation by glucose with a linear fit, y =-0·0895x + 230·99(R² = 0·7676, p = 0·013). The glycation by fructose was greater than that of glucose with stronger inhibitory effect by zinc in fructose-glycation (t= -5.8, p=0.002). Addition of zinc significantly decreased fluorescence as seen in Zn + FA or Zn + AA sets as compared to sets of FA alone (p=0.00056) or AA alone (p=0.037). The fluorescence for dityrosine and AGE had a correlation of 0.897 (p<0.01). The data from fluorescence, UV, and FTIR spectra collectively suggested inhibitory effect of zinc in BSA glycation alone or in presence of FA and AA, showing new dimension for the protective action of zinc in hyperglycemic conditions.

This study investigated the antioxidant activity of methanol (MeOH) and water extracts from roots of Cirsium japonicumin vitro. MeOH extract showed a stronger free radical scavenging activity than water extract. However, both of extracts showed a concentration dependent hydroxyl radical scavenging activity, reducing power and metal chelating ability. MeOH extract had greater phenolic and flavonoid contents than water extract. The antidiabetic activity of these two extracts was evaluated by the alpha-glucosidase inhibition assay. The water extract showed a considerable alpha-glucosidase inhibitory activity. To our knowledge, this may be the first time to report the antioxidant and antidiabetic activities in Cirsium japonicum roots.

Many new polyisoprenylated benzophenones with a bicyclo[3.3.1]-nonane-2,4,9-trione core structure have been isolated from plants in the Clusiaceae family, and their potent biological properties have been the subject of several studies. This review summarizes the biological activities reported for these secondary metabolites including cytotoxic, antimicrobial, antioxidant, and anti-inflammatory activities. Our efforts during the past years have foremost been directed towards isolating new polyisoprenylated benzophenones, as well as understanding the possible target and mechanism of action through which these compounds arrest cancer cells and inhibit the progression of the cell-cycle. The transcription of genes is affected in cancer cells treated with polyisoprenylated benzophenones; the oncogene c-Myb is down-regulated and endoplasmatic stress genes XBP1, ATF4, and DDIT3/CHOP are turned on. Consequently, the expression of iNOS and cell cycle regulators such as cyclin D and E are reduced. Evidence presented by independent investigators suggests that polyisoprenylated benzophenones affect the mediators in the Akt/mTOR stress pathway, although the specific target remains to be discovered. In addition, benzophenones isolated from plants display high antioxidant capacity and protect cells from oxidative stress and the formation of ROS involved during the inflammatory process. Since antiviral activity was initially reported for guttiferone A, potent synthetic analogues have been developed as effective new non-nucleoside reverse transcriptase inhibitors (NNRTI) to treat drug resistant HIV-1. In addition, benzophenones exert antimicrobial effects particularly against MRSA. The structure-activity relationships of polyisoprenylated benzophenones from natural sources and those of synthetic analogues are included in this review. Absorption, metabolism, and elimination of benzophenones are also discussed.

Garcinol, harvested from Garcinia indica, has traditionally been used in tropical regions and appreciated for centuries; however its biological properties are only beginning to be elucidated. There is ample data to suggest potent antioxidant properties of this compound which have been used to explain most of its observed biological activities. However, emerging evidence suggests that garcinol could be useful as an anti-cancer agent, and it is increasingly being realized that garcinol is a pleiotropic agent capable of modulating key regulatory cell signaling pathways. Here we have summarized the progress of our current research knowledge on garcinol and its observed biological activities. We have also provided an explanation of observed properties based on its chemical structure and provided an insight into the structure and properties of chalcones, the precursors of garcinol. The available data is promising but more detailed investigations into the various properties of this compound, particularly its anti-cancer activity are urgently needed, and it is our hope that this review will stimulate further research for elucidating and appreciating the value of this nature's wonder agent.

The effects of garcinol and guttiferone K, two polyisoprenylated benzophenones occurring a food plant called Garcinia cambogia, on oxidative/nitrative protein damage (determined by parameters such as levels of protein carbonyl groups and nitrotyrosine residues) in human blood platelets and plasma after treatment with peroxynitrite (ONOO(-)) were studied in vitro. We also investigated the effects of garcinol and guttiferone K on lipid peroxidation in blood platelets and plasma induced by ONOO(-)(100 microM). Exposure of blood platelets or plasma to peroxynitrite (100 microM) resulted in an increased level of carbonyl groups and nitrotyrosine residues in proteins, and an increase of lipid peroxidation measured by the level of thiobarbituric acid reactive species (TBARS). In the presence of garcinol and guttiferone K (0.1-25 microg/ml), a distinct reduction in the formation of carbonyl groups in plasma and platelet proteins together with the decrease of TBARS caused by 100 microM peroxynitrite, was observed. However, garcinol and guttiferone K did not inhibit plasma and platelet protein nitration induced by peroxynitrite. Polyisoprenylated benzophenones present in human diet such as garcinol or guttiferone K in vitro have protective effects against lipid and protein oxidation and may have some promising effects in vivo because they are good antioxidants in the tested models in vitro. Garcinol and guttiferone K can be also useful as protecting factors against diseases associated with oxidative stress.

Garcinol, a polyisoprenylated benzophenone derivative, was purified from Garcinia indica fruit rind, and its free radical scavenging activity was studied using electron spin resonance (ESR) spectrometry. In the hypoxanthine/xanthine oxidase system, emulsified garcinol suppressed superoxide anion to almost the same extent as DL-alpha-tocopherol by weight. In the Fenton reaction system, garcinol also suppressed hydroxyl radical more strongly than DL-alpha-tocopherol. In the H(2)O(2)/NaOH/DMSO system, garcinol suppressed superoxide anion, hydroxyl radical, and methyl radical. It was thus confirmed that this derivative is a potent free radical scavenger and able to scavenge both hydrophilic and hydrophobic ones including reactive oxygen species. Orally administered garcinol prevented acute ulceration in rats induced by indomethacin and water immersion stress caused by radical formation. These results suggested garcinol might have potential as a free radical scavenger and clinical application as an antiulcer drug.

An H(2)O(2)/NaOH/DMSO system has been developed for the formation of three free radicals, and the application of the system was examined with the antioxidants ascorbic acid and tocopherol. Superoxide anion, hydroxyl radical,and methyl radical are simultaneously generated in this system. The scavenging activity of ascorbic acid and tocopherol for these radicals was estimated by 5, 5'-dimethyl-1-pyrroline-N-oxide spin trapping electron spin resonance. Both water-soluble and oil-soluble antioxidants could be evaluated by using this system. Ascorbic acid specifically inhibited the superoxide anion and hydroxyl radical, whereas tocopherol suppressed the methyl radical.

The scavenging effects of grape seed extract (GSE) on free radicals formed in an H(2)O(2)/NaOH/DMSO system were examined using a spin-trapping electron spin resonance (ESR) method and compared with other natural antioxidants, ascorbic acid, dl-alpha-tocopherol, and beta-carotene. GSE reduced greatly the ESR signal intensity of superoxide radical-5,5-dimethyl-1-pyrroline-N-oxide (DMPO) adducts. GSE also exhibited weak scavenging activity on hydroxyl radical and a little scavenging activity on methyl radical. Ascorbic acid exhibited strong superoxide and hydroxyl radical scavenging activities, but it increased the amount of methyl radical at high concentration. dl-alpha-Tocopherol reduced the amount of superoxide anion, especially the amount of methyl radical. However, it slightly reduced the amount of hydroxyl radical. beta-Carotene reduced the amount of hydroxyl radical and methyl radical, but it also slightly reduced superoxide anion. In the case of combination use of beta-carotene and dl-alpha-tocopherol, all radical species were suppressed. Combination of GSE and dl-alpha-tocopherol also could reduce all radical species. beta-Carotene and dl-alpha-tocopherol could reduce the methyl radical formation induced by ascorbic acid.

Bisphenol A diglycidyl ether (BADGE) is an epoxy resin monomer. Epoxy-based solution coatings are used in many applications as additives for a variety of plastic coatings in food packaging. It is well known that unreacted BADGE can migrate from epoxy-based packing materials into foods. Not only BADGE but also its derivatives can easily migrate into foods and it is likely that we intake BADGE and its derivatives through food or drink. Recently, endocrine disrupting chemicals (EDCs) have attracted attention because they have been shown to affect reproduction in wildlife. The estrogenic activity of BADGE derivatives has not previously been investigated. Therefore, we investigated the estrogenic activity of the BADGE derivatives, dihydrolysed BADGE (BADGE-4OH) and chlorohydroxy BADGE (BADGE-2Cl), using breast cancer cell (T47D) proliferation assay and estrogen receptor (ER)(alpha) binding assay. These chemicals exhibited T47D cell proliferation at concentrations of 10(-14)-10(-4) M. However, these chemicals did not bind to ER (alpha) in the binding assay.

A sensitive HPLC method for determination of bisphenol A (BPA) in plasma samples using 4-(4,5-diphenyl-1H-imidazol2-yl)benzoyl chloride (DIB-Cl) as a fluorescence labeling reagent was developed. The fluorescence labeling reaction was completed within 10 min at room temperature. DIB-Cl reacts with the phenolic hydroxyl group of BPA in the presence of triethylamine (TEA). The DIB-Cl derivative of BPA (DIB-BPA) was separated within 30 min with an ODS column using acetonitrile-water (90:10, v/v) as the isocratic eluent. Calibration graphs were linear over the range of 1.0-100 ng/ml (r=0.999). The detection limit of DIB-BPA was 0.05 ng/ml (2.5 pg) at a signal-to-noise ratio of 3. The relative standard deviations (RSDs) of the method for between-run were 1.0-5.0%. The analytical recoveries of known amounts (1.0 and 100 ng/ml) of BPA-spiked rabbit plasma were around 95%.

We report a new approach for assessing human exposure to bisphenol A (BPA) by measuring BPA in urine after enzymatic deglucuronidation. This method involves addition of (13)C(12)-labeled BPA, enzymatic deconjugation, solid-phase extraction, and derivatization with pentafluorobenzyl bromide. The product of the derivatization is separated by gas chromatography followed by mass spectrometric detection using negative chemical ionization and selected ion monitoring. Using this analysis method, urine samples fortified with both a constant level of labeled BPA and a range of unlabeled BPA levels (0.27-10.6 ng/ml) demonstrated constant percentage recovery. In addition, a range of urine sample volumes (0.25-10.0 ml) with constant amounts of added internal standard produced a linear response (r(2)=0.99). The method limit of detection was 0.12 ng/ml. This method was validated by duplicate analyses using gas chromatography coupled to a high-resolution mass spectrometer.

Migration of 4-nonylphenol (NP) from polyvinyl chloride (PVC) films for food packaging into food simulants and foods has been studied in domestic applications such as wrapping of food and reheating in a microwave oven. The migration of NP from the PVC films was determined by high-performance liquid chromatography with electrochemical coulometric-array detection (LC/ED). Twelve PVC films intended for commercial use and ten for domestic applications (total: 22 samples) were analysed. Some of the PVC films (two home-use and ten retail-use) contained NP at concentrations of between 500 and 3300 microg/g. Migration of NP from the films was influenced by the test conditions (n-heptane at 25 degrees C for 60 min, distilled water at 60 degrees C for 30 min and 4% acetic acid at 60 degrees C for 30 min). The amount of NP migrating from the PVC films into n-heptane (0.33-1.6 microg/cm2) was higher than the amount migrating into distilled water or 4% acetic acid (up to 9.7 ng/cm2) for the 11 films in which NP was detected. Up to 0.23% of the NP migrated into distilled water and 4% acetic acid and up to 62.5% into n-heptane. In addition, we investigated NP migration into cooked rice samples wrapped in PVC film. Using spiked samples the method gave an average recovery of 83.7%(n = 5) with a standard deviation of 2.5%. Migration of NP ranged from not detectable (< 1.0 ng/g) to 410.0 ng/g by reheating samples in a microwave oven for 1 min and from not detectable to 76.5 ng/g by keeping samples at room temperature for 30 min.

Alkylphenols can affect human health because they disrupt the endocrine system. In this study, an analytical method for determining trace amounts of 4-nonylphenol (NP) and 4-octylphenol (OP) in human blood samples was developed. Reversed-phase HPLC with multi-electrode electrochemical coulometric-array detection was used for the determination of NP and OP in plasma and serum samples prepared with a solid-phase extraction method. The separation was achieved using an isocratic mobile phase of 0.7% phosphoric acid-acetonitrile with a C18 reversed phase column. The detection limits of NP and OP were 1.0 and 0.5 ng ml-1, respectively. The recoveries of NP and OP added to human plasma samples were above 70.0% with a relative standard deviation of less than 15.5%. The method was found to be applicable to the determination of NP and OP in various human blood samples such as serum and plasma.

A simple and sensitive method using high-performance liquid chromatography with multi-electrode electrochemical detection (HPLC-ED) including a coulometric array of four electrochemical sensors has been developed for the determination of bisphenol A in water and human serum. For good separation and detection of bisphenol A, a CAPCELL PAK UG 120 C18 reversed-phase column and a mobile phase consisting of 0.3% phosphoric acid-acetonitrile (60:40) were used. The detection limit obtained by the HPLC-ED method was 0.01 ng/ml (0.5 pg), which was more than 3000-times higher than the detection limit obtained by the ultraviolet (UV) method, and more than 200-times higher than the detection limit obtained by the fluorescence (FL) method. Bisphenol A in water and serum samples was pretreated by solid-phase extraction (SPE) after removing possible contamination derived from a plastic SPE cartridges and water used for the pretreatment. A trace amount (ND approximately 0.013 ng/ml) of bisphenol A was detected from the parts of cartridges (filtration column, sorbent bed and frits) by extraction with methanol, and it was completely removed by washing with at least 15 ml of methanol in the operation process. The concentrations of bisphenol A in tap water and Milli-Q-purified water were found to be 0.01 and 0.02 ng/ml, respectively. For that reason, bisphenol A-free water was made to trap bisphenol A in water using an Empore disk. In every pretreatment, SPE methods using bisphenol A-free water and washing with 15 ml of methanol were done in water and serum samples. The yields obtained from the recovery tests using water to which 0.5 or 0.05 ng/ml of bisphenol A was added were 83.8 to 98.2%, and the RSDs were 3.4 to 6.1%, respectively. The yields obtained from the recovery tests by OASIS HLB using serum to which 1.0 ng/ml or 0.1 ng/ml of bisphenol A was added were 79.0% and 87.3%, and the RSDs were 5.1% and 13.5%, respectively. The limits of quantification in water and serum sample were 0.01 ng/ml and 0.05 ng/ml, respectively. The method was applied to the determination of bisphenol A in healthy human serum sample, and the obtained detection was 0.32 ng/ml. From these results, the HPLC-ED method should be the most useful in the determination of bisphenol A at low concentration levels in water and biological samples.

The modifying effects of dietary feeding of a polyisoprenylated benzophenone, garcinol, isolated from Garcinia indica fruit rind on the development of azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) were investigated in male F344 rats. We also assessed the effects of garcinol on proliferating cell nuclear antigen (PCNA) index in ACF and activities of detoxifying enzymes of glutathione S-transferase (GST) and quinone reductase (QR) in liver. In addition, we examined the effects of garcinol on 12-O-tetradecanoylphorbol-13-acetate-induced O(2)(-) generation in differentiated human promyelocytic HL-60 cells and lipopolysaccharide (LPS)- and interferon (IFN)-gamma-induced nitric oxide (NO) generation in mouse macrophage RAW 264.7 cells. Western blotting analysis of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression was done in LPS- and IFN-gamma-treated mouse macrophage RAW 264.7 cells. Rats were given subcutaneous injections of AOM (15 mg/kg body wt) once a week for 3 weeks to induce ACF. They also received the experimental diet containing 0.01 or 0.05% garcinol for 5 weeks, starting 1 week before the first dosing of AOM. AOM exposure produced 97 +/- 15 ACF/rat at the end of the study (week 5). Dietary administration of garcinol caused significant reduction in the frequency of ACF: 72 +/- 15 (26% reduction, P < 0.01) at a dose of 0.01% and 58 +/- 8 (40% reduction, P < 0.001) at a dose of 0.05%. Garcinol administration significantly lowered PCNA index in ACF. Feeding of garcinol significantly elevated liver GST and QR activities. In addition, garcinol could suppress O(2)(-) and NO generation and expression of iNOS and COX-2 proteins. These findings might suggest possible chemopreventive ability of garcinol, through induction of liver GST and QR, inhibition of O(2)(-) and NO generation and/or suppression of iNOS and COX-2 expression, on colon tumorigenesis.

The purpose of the current study was to determine the anti-obesity and anti-inflammatory effects of an extract of purple sweet potatoes (PSPs) on 3T3-L1 adipocytes. For this purpose, differentiated 3T3-L1 adipocytes were treated with a PSP extract at concentrations of 1,000, 2,000, and 3,000 μg/mL for 24 hours. Then, we measured the changes in the sizes of the adipocytes, the secretion of leptin, and the mRNA/protein expression of lipogenic, inflammatory, and lipolytic factors after the treatment with the PSP extract. The PSP extract diminished leptin secretion, indicating that growth of fat droplets was suppressed. The extract also suppressed the expression of mRNAs of lipogenic and inflammatory factors and promoted lipolytic action. The antioxidative activity of the PSP extract was also measured using three different in vitro methods: 1,1-diphenyl-2-picrylhydrazyl free radical scavenging activity, ferric reducing ability potential assay, and chelating activity of transition metal ions. Taken together, our study shows that PSP extract has antilipogenic, anti-inflammatory, and lipolytic effects on adipocytes and has radical scavenging and reducing activity.

In the present study, the seeds and fruit rind of six plants of the Fabaceae family were selected to evaluate their potential as antioxidant and antibacterial agents. The dried powders were individually extracted with various organic solvents by the cold percolation method, were evaluated for antibacterial activity and methanol extracts used for antioxidant activities. Total phenol, protein and sugar contents were also measured. Antioxidant activities were measured by DPPH free radical scavenging activity, superoxide anion radical scavenging activity and reducing capacity assessment. Antibacterial activity was measured by the agar well diffusion method against four Gram positive and four Gram negative bacteria. The methanol extract of the fruit rind of C. indica showed the maximum DPPH free radical scavenging activity, superoxide anion radical scavenging activity, a high reducing capacity assessment and also had the highest total phenol content. There was a direct correlation between the phenol content and the antioxidant activity. The antibacterial activity of all the extracts was more pronounced on Gram positive bacteria than on Gram negative bacteria. Thus, the fruit rind of C. indica showed the best antioxidant and antibacterial activities.

Rhizophora mangle (L.) produce a variety of substances that possesses pharmacological actions. Although it shown antioxidant properties in some assays, there is no available information about its effect on some free radical species. So the objective of the present research is to evaluate the DPPH radical and superoxide anion scavenging properties of R. mangle extract and its polyphenol fraction. Rhizophora mangle (L.) bark aqueous extract and its major constituent, polyphenols fraction, were investigated for their antioxidant activities employing 2 in vitro assay systems: 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide anion radicals scavenging. IC(50) for DPPH radical-scavenging activity was 6.7 µg tannins/mL for extract and 7.6 µg tannins/mL for polyphenolic fraction. The extract showed better activity than its fraction (P < 0.05) in the DPPH radicals reducing power. Polyphenolic fraction exhibited better superoxide anion scavenging ability (IC(50)= 21.6 µg tannins/mL) than the extract (IC(50)= 31.9 µg tannins/mL). Antioxidant activities of both samples increased with the rise of tannins concentration. The comparison of regression lines showed significant differences (P < 0.05) between extract and its polyphenolic fraction in both assays, indicating that extract was more effective in DPPH radical scavenging than its fraction at tannin concentrations below the crossing point of both lines, while that fraction was more effective than extract inhibiting the superoxide anions generation. R. mangle aqueous extract showed a potent antioxidant activity, achieved by the scavenging ability observed against DPPH radicals and superoxide anions. Regarding its polyphenolic composition, the antioxidant effects observed in this study are due, most probably, to the presence of polyphenolic compounds.

The phytochemical composition of carotenoids, tocopherols, free sugars, organic acids, L-ascorbic acid, capsaicinoids, and flavonoids in green and red paprika (GP and RP), and paprika leaves (PL) cultivated in Korea were analyzed. The ethanolic extracts of GP, RP, and PL were obtained with 80% ethanol, and their antioxidative activities were determined by measuring their ABTS and DPPH radical scavenging activities. RP showed the highest contents of capsanthin (58.33 ± 3.91 mg/100 g dry weight) and L-ascorbic acid (1987.25 ± 19.64 mg/100 g dry weight), and main compounds of PL were lutein, chlorophyll, and γ-tocopherol (96.91 ± 14.58, 2136.71 ± 21.11, and 723.49 ± 54.10 mg/100 g dry weight, respectively). RP showed the strongest antioxidant activity (IC(50)= 55.23 ± 6.77 μg/mL in a 2, 2'-azino-di-[3-ethylbenzthiazoline sulphonate] assay and 150.40 ± 8.07 μg/mL in a 2, 2-diphenyl-2-picrylhydrazyl assay), and the antioxidant activity of PL was higher than β-carotene but lower than RP. The results indicate that the amounts of capsanthin and L-ascorbic acid in RP correlate well with antioxidant activity. PL, which has various phytochemicals such as lutein, chlorophyll, and γ-tocopherol, might be used in nutraceuticals and pharmaceuticals for improving human health.

Taking bovine serum albumin (BSA) as typical molecules, the sonodynamic damage of protein in the presence of Levofloxacin (LVFX) and its mechanism were studied by fluorescence and UV-vis spectra. Various influencing factors such as ultrasonic irradiation time, pH value, ionic strength and solution temperature on the damage of BSA were also discussed. The results showed that ultrasound can enhance the damage of LVFX on BSA. The damage degree of BSA was aggravated with the increase of ultrasonic irradiation time, solution temperature and ionic strength, whereas decreased with the increase of solution pH value. Furthermore, the reactive oxygen species (ROS) in reaction system were studied by oxidation and extraction photometry. Experimental results showed that the amounts of superoxide anion radical (.O(2)(-)) and hydroxyl radical (.OH) were significantly more than that of singlet oxygen ((1)O(2)) in the presence of LVFX under ultrasonic irradiation.

Alcalase-treated zein hydrolysate (ZH) was separated by gel filtration, ultrafiltration, and reversed-phase HPLC, and the scavenging activities for 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS*+), 1,1-diphenyl-2-picrylhydrazyl (DPPH*), and superoxide anion (O2*-) radicals of different peptide fractions were measured to establish the antioxidant efficacy. Results showed that the ability to stabilize water-soluble free radicals (ABTS*+) by ZH components was insensitive to the peptide size, whereas that against ethanol-soluble free radicals (DPPH*) and O2*- was molecular weight dependent. Antioxidative peptides of <1 kDa were further separated by HPLC into 30 components, of which those with great hydrophobicity exhibited strong DPPH* and O2*- scavenging ability and those with intermediate hydrophobicity displayed the maximum ABTS*+ scavenging activity. Two dominant components (fractions 8 and 17) were further purified and identified by LC-PDA-ESI-MS to be Tyr-Ala and Leu-Met-Cys-His, respectively. The results demonstrated that the free radical scavenging activity of ZH depended on the radical species and was strongly related to the molecular weight and hydrophobicity of the constituting peptides.

AIMS To evaluate the free radical-scavenging capacity of Lactobacillus fermentum and its effects on antioxidant enzyme levels in finishing pigs. METHODS AND RESULTS The free radical-scavenging activity of Lact. fermentum was analysed in vitro. The tested Lactobacillus showed a high scavenging ability against DPPH (1,1-diphenyl-2-picrylhydrazyl), superoxide and hydroxyl radicals which was dose dependent. Subsequently, 108 crossbred pigs weighing 20.67 BW, were allotted to dietary treatments including a basal diet or the basal diet supplemented with either aureomycin or 10.2 x 10(7) Lact. fermentum CFU g(-1) diet. Supplementation of Lact. fermentum increased total antioxidant capacity (P < 0.01) in serum from 50 kg pigs, while serum superoxide dismutase (P = 0.01) and glutathione peroxidase (P < 0.01) increased, and malondialdehyde levels decreased (P < 0.01) in 90 kg pigs. Hepatic catalase (P = 0.04), muscle superoxide dismutase (P < 0.01) and copper-zinc-superoxide dismutase were enhanced (P = 0.01), whereas malondialdehyde levels were reduced (P = 0.05) by Lact. fermentum. CONCLUSIONS The free radical-scavenging capacity of Lact. fermentum was dose dependent and its supplementation improved the antioxidant status of pigs. SIGNIFICANCE AND IMPACT OF THE STUDY Lactobacillus fermentum could be used to alleviate oxidative stress and increase pig performance and improve pork quality.

The antioxidative activities of water extract (WE) and crude hot-water soluble polysaccharide (PS) from Ficus carica L. fruit were investigated using various assays in vitro, including scavenging abilities on DPPH, superoxide and hydroxyl radicals and reducing power. The immunity activities of PS were evaluated using the carbon clearance test and serum hemolysin analysis in mice. In addition, total phenolics and flavonoids contents were also determined. Both WE and PS have notable scavenging activities on DPPH with the EC(50) values of 0.72 and 0.61 mg/ml, respectively. The PS showed higher scavenging activity than WE on superoxide radical (EC(50), 0.95 mg/ml) and hydroxyl anion radical (scavenging rate 43.4% at concentration of 4 mg/ml). The PS (500 mg/kg) also has a significant increase in the clearance rate of carbon particles and serum hemolysin level of normal mice. The results indicate that both WE and PS might be applicable in healthy medicine and food industry.

To identify the potential of green leafy vegetables (GLV) as antioxidants, methanolic extracts of Amaranthus sp., Centella asiatica, Murraya koenigii and Trigonella foenum graecum were studied for their antioxidant activity in different systems at multiple concentrations. Total antioxidant activity assessed by phosphomolybdenum method, free radical scavenging activity by 1,1-diphenly-2-picryl hydrazyl (DPPH), reducing power and ferrous ion chelating activity were determined. The GLV were analyzed for ascorbic acid, total and beta-carotene and total polyphenol contents. The ascorbic acid, total carotene, beta-carotene and total phenolic content (tannic acid equivalents) of the GLV ranged between 15.18-101.36, 34.78-64.51, 4.23-8.84 and 150.0-387.50 mg/100 g GLV, respectively. The extracts were found to have significantly different levels of antioxidant activities in the systems tested. The total antioxidant activity was highest in Murraya koenigii (2,691.78 micromol of ascorbic acid/g sample) and least in Centella asiatica (623.78 micromol of ascorbic acid/g sample). The extract concentration causing 50% inhibition of DPPH (IC50) was determined (M. koenigii<C.asiatica<Amaranthus sp.<T. graecum). The maximum DPPH scavenging activity and reducing power was exhibited by Murraya koenigii. Multiple regression analysis showed that the relationship of total antioxidant activity, free radical scavenging activity, and reducing power with polyphenol and total and beta-carotene was highly significant.

Model Maillard reaction products were generated by heating glucose-lysine mixtures (GL) at 150 degrees C for different times (15, 30, 60, and 90 min). Samples were characterized by free lysine, browning, and UV-visible spectra and assessed for antioxidant properties, metal chelating ability, and effects on mineral absorption across Caco-2 monolayers. It was found that the capacity to retard lipid peroxidation in a model linoleic acid emulsion system increased with heating time up to 60 min and then leveled off, whereas the scavenging activity toward 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radicals increased in early periods of the reaction (15 and 30 min of heating) and decreased thereafter. The iron binding affinity of the different samples was not correlated with antioxidant properties, and iron transport in Caco-2 cells was unchanged between samples. On the contrary, copper chelating activity showed significant correlation with free radical scavenging activity and with copper absorption across intestinal cells. It can be concluded that severe heat treatment of GL mixtures maintained the ability to reduce lipid peroxidation but decreased the free radical scavenging activity. Moreover, antiradical activity, copper chelation ability, and positive effects on copper absorption were correlated and associated to compounds formed at early stages of the Maillard reaction.