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Summary Developing functional food products requires a structured approach for screening potential ingredients. Using the development of a product with antioxidant properties as a model, eight fruit juice concentrates were screened, including New Zealand fruits. The total phenolic content and the antioxidant capacity, using chemical-based assays (ferric reducing antioxidant power and oxygen radical absorbance capacity assays), were determined. The potential to protect against oxidative stress in Jurkat cells was also determined. Results indicate that the total phenolic content and antioxidant capacity in chemical-based assays were strongly correlated. However, the total phenolic content did not dictate the level of protection from oxidative stress in this cell-based model, suggesting that the type of phenolic compounds present and interactions between them may be important, although the activity of other compounds, such as vitamin C or carotenoids, may also be implicated. This model for screening ingredients may be used to design synergistic combinations of ingredients, and results may provide supportive evidence in the development of functional foods for this and other health targets.

The cytoprotective effects of five flavonoids (quercetin, rutin, catechin, kaempferol and morin) and four hydroxycinnamic acids (caffeic, ferulic, sinapic and chlorogenic acids) were evaluated by the degree of protection they provided against H2O2-induced damage to human neuroblastoma SH-SY5Y cells. All compounds exhibited protection against H2O2-mediated cytotoxicity in a dose dependent manner. The concentration required to give a 50% reduction in cell death (EC50 value) were derived from their dose–response relationships. The cytoprotective activities of these phenolic compounds in the order of quercetin > caffeic acid > rutin > chlorogenic acid > catechin > ferulic acid > sinapic acid > morin > kaempferol. The EC50 values of the phenolic compounds were strongly related to their chemical structures. The EC50 values were compared with the antioxidant activities as determined by five different chemically based antioxidant assays [2,2-azinobis (3-ethyl-benzothiazoline-6-sulfonic acid (ABTS), 1,1-diphenyl-2-picrylhydrazyl (DPPH), oxygen radical absorbance capacity (ORAC), ferric reducing antioxidant power (FRAP) and lipid peroxidation inhibition capacity (LPIC)]. The ability of these phenolic compounds to protect from H2O2-induced SH-SY5Y cell death correlated (r 2 = 0.85) with their determined LPIC values and weakly (r 2 = 0.44) with their ABTS activities. There was no correlation between EC50 values and ORAC, FRAP or DPPH activities. The cytoprotection assay is a more biologically relevant measurement than the chemically defined antioxidant activity assays because it uses human cells as a substrate and therefore accounts for some aspects of uptake, metabolism and location of antioxidant compounds within cells.

Purpose: To determine the antitumor activity of the anti–mesothelin immunotoxin SS1P in combination with gemcitabine against mesothelin-expressing tumor xenografts. Experimental Design: The in vitro activity of SS1P in combination with gemcitabine against the mesothelin-expressing cell line A431/K5 was evaluated using cytotoxicity and apoptosis assays. The antitumor activity of this combination was evaluated in nude mice bearing A431/K5 tumor xenografts. Tumor-bearing mice were treated with different doses and schedules of gemcitabine alone, SS1P alone (0.2 mg/kg i.v. every other day × three doses), or with both agents together, and tumor volumes were measured over time. Results: In vitro studies failed to show the synergy of SS1P plus gemcitabine against the mesothelin-expressing A431/K5 cells. In contrast, in the in vivo setting, there was a marked synergy when SS1P was combined with gemcitabine for the treatment of mesothelin-expressing tumor xenografts. This synergy was present using different doses and schedules of gemcitabine administration. In mice treated with fractionated doses of gemcitabine in combination with SS1P, complete tumor regression was observed in all mice and was long-lasting in 60% of the animals. Also, this antitumor activity was specific to SS1P because HA22, an immunotoxin targeting CD22 not expressed on A431/K5 cells, did not increase the efficacy of gemcitabine. Conclusions: SS1P in combination with gemcitabine results in marked antitumor activity against mesothelin-expressing tumors. This combination could be potentially useful for the treatment of human cancers that express mesothelin and are responsive to gemcitabine therapy.

To simulate the effects of digestion and metabolism on the survival of different polyphenolic compounds, extracts of blueberry and apple were deglycosylated by acid hydrolysis, followed by enzymic glucuronidation under neutral conditions, yielding approximately 5% overall recovery of polyphenolics. The major polyphenolics before and after the treatment were compared, to estimate which species are likely to be present in the intestinal lumen, undegraded and available for absorption, after consumption of the fruit. Whereas blueberry extract consisted predominantly of anthocyanins, epicatechin and caffeoyl quinate esters, the major components of the treated extract were quercetin glucuronides and (unglucuronidated) caffeoyl quinates, with only traces of anthocyanidin derivatives. In apple extract, compositional changes were less marked, but caffeoyl quinates, procyanidins and quercetin were enriched at the expense of caffeic acid, epicatechin and catechin. Hydrophobic compounds like phloretin and quercetin were extensively glucuronidated, whereas caffeic acid and caffeoyl quinate were not. These results suggest that the major polyphenolic components of a fruit are not necessarily the most important contributors to any health benefits because the polyphenolic composition in the intestinal lumen and consequently, in the circulation, may be considerably different.

Background: Although ethanol addiction is believed to be mediated by the mesolimbic dopamine system, originating from the ventral tegmental area (VTA), how acute ethanol increases the activity of VTA dopaminergic (DA) neurons remains unclear. Method: Patch-clamp recordings of spontaneous firings of DA and GABAergic neurons in the VTA in acute midbrain slices from rats. Results: Ethanol (20–80 mM) excites DA neurons, and more potently depresses firing of local GABAergic neurons. The ethanol-induced excitation of DA neurons is considerably attenuated by DAMGO (Tyr-d-Ala-Gly-N-Me-Phe-Gly-ol enkephalin), a μ-opioid agonist that suppresses firing of GABAergic neurons, or by naloxone, a general opioid antagonist. The ongoing opioid-induced facilitation of DA cell firing (revealed by naloxone) is enhanced by ethanol, probably by an increase in opioid release or action. Conclusion: Ethanol excites VTA DA neurons at least partly by increasing ongoing opioid-mediated suppression of local GABAergic inhibition. This indirect mechanism may contribute significantly to the positively reinforcing properties of ethanol.

Phenolic acid esters have various potential uses as functional food ingredients, e.g., as antioxidants and preservative agents. Combinatorial mixtures of phenolic acid esters were generated by mixing several different alcohol and acid substrates in various combinations and esterifying enzymically. It was shown to be feasible to identify all the combinatorial products using LCMS and to separate the individual components by HPLC for separate bioassay. Ester mixtures were tested, in comparison with un-esterified substrate mixtures, for anti-microbial effects and found to be active against strains of human gut bacteria. Esterification did not increase the strength of the activity, but significant synergies, both positive and negative, between the components of different ester mixtures were found. Isolated components of a mixture had much less activity than the mixture itself. In tests for protection of cultured cells from H2O2-induced oxidative damage and consequent apoptosis, esterification could increase the strength of protection, possibly because of increased bioavailability of the more membrane-soluble hydrophobic esters, compared with the water-soluble substrates. The combinatorial enzymic approach to generation of synergistic mixtures of phenolic acid esters has potential to find synergistic combinations that would be missed by testing individual compounds and combining only the active compounds.

To explore the effects of exogenous carbon monoxide-releasing molecules 2 (CORM-2) on LPS-induced abnormal activation of platelets in peripheral blood of healthy human donors and its possible molecular mechanism.Venous blood samples were collected from a healthy volunteer, and platelet-rich plasma (PRP) from the blood were isolated by differential centrifugation. The PRP was subpackaged into siliconized test tubes and then divided into control group, LPS group, inactive CORM-2 (iCORM-2) group, 10 µmol/L CORM-2 group, and 50 µmol/L CORM-2 group according to the random number table, with 3 tubes in each group. The PRP in control group did not receive any treatment. The PRP in LPS group received LPS (20 mL, 10 µg/mL) stimulation, and the PRP in iCORM-2 group, 10 µmol/L CORM-2 group, and 50 µmol/L CORM-2 group underwent the same LPS stimulation and treatment of 50 µmol/L iCORM-2, 10 µmol/L CORM-2, and 50 µmol/L CORM-2, respectively, with the dosage of 20 mL. After being cultured for 30 min, the platelet adhesion rate was determined by glass bottle method, the number of platelet spreading on fibrinogen was determined with immunofluorescent method, and the platelet aggregation rate was measured by turbidimetric method. The platelet poor plasma (PPP) was prepared from PRP, the levels of ATP in PPP and platelets were determined by chemical fluorescein method. The expressions of platelet glycoprotein I bα (GPIbα) and GPVI were analyzed by flow cytometer. The expressions of glycogen synthase kinase 3β (GSK-3β) and phosphorylated GSK-3β were determined by Western blotting and immunoprecipitation, respectively. Measurement of the above indices was repeated for 3 times. Data were processed with one-way analysis of variance and SNK test.Compared with those in control group, the platelet adhesion rates, numbers of platelets spreading on fibrinogen, platelet aggregation rates, expressions of GPIbα and GPVI in PRP, levels of ATP in PPP in LPS and iCORM-2 groups were significantly increased, while levels of ATP in platelets were significantly decreased (with P values below 0.05). Compared with those in LPS group, the former 7 indices in iCORM-2 group showed no significant differences (with P values above 0.05), while the levels of ATP in platelets in the 10 µmol/L CORM-2 and 50 µmol/L CORM-2 groups were significantly increased, and the other 6 indices in 10 µmol/L CORM-2 and 50 µmol/L CORM-2 groups were significantly decreased (with P values below 0.05). The expression levels of GSK-3β of the platelets in PRP in control, LPS, iCORM-2, 10 µmol/L CORM-2, and 50 µmol/L CORM-2 groups were 0.550 ± 0.060, 1.437 ± 0.214, 1.210 ± 0.108, 0.720 ± 0.010, and 0.670 ± 0.010, respectively, and the expression levels of the phosphorylated GSK-3β of the platelets in PRP in the above 5 groups were 0.950 ± 0.070, 1.607 ± 0.121, 1.420 ± 0.040, 1.167 ± 0.015, and 0.513 ± 0.122, respectively. Compared with those in control group, both the expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP in LPS and iCORM-2 groups were significantly increased (with P values below 0.05). The expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP between LPS group and iCORM-2 group were similar (with P values above 0.05). The expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP in 10 µmol/L CORM-2 and 50 µmol/L CORM-2 groups were significantly decreased compared with those in LPS group (with P values below 0.05).LPS stimulation can abnormally activate the platelets in peripheral blood of healthy human, but the abnormal activation can be inhibited by CORM-2 intervention, and the mechanism of the latter may involve the phosphorylation of GSK-3β mediated by GP.

Background The development and evaluation of new therapeutic approaches for malignant mesothelioma has been sparse due, in part, to lack of suitable tumor models. Methods We established primary mesothelioma cultures from pleural and ascitic fluids of five patients with advanced mesothelioma. Electron microscopy and immunohistochemistry (IHC) confirmed their mesothelial origin. Patient derived xenografts were generated by injecting the cells in nude or SCID mice, and malignant potential of the cells was analyzed by soft agar colony assay. Molecular profiles of the primary patient tumors, early passage cell cultures, and patient derived xenografts were assessed using mutational analysis, fluorescence in situ hybridization (FISH) analysis and IHC. Results Primary cultures from all five tumors exhibited morphologic and IHC features consistent to those of mesothelioma cells. Mutations of BAP1 and CDKN2A were each detected in four tumors. BAP1 mutation was associated with the lack of expression of BAP1 protein. Three cell cultures, all of which were derived from BAP1 mutant primary tumors, exhibited anchorage independent growth and also formed tumors in mice, suggesting that BAP1 loss may enhance tumor growth in vivo. Both early passage cell cultures and mouse xenograft tumors harbored BAP1 mutations and CDKN2A deletions identical to those found in the corresponding primary patient tumors. Conclusions The mesothelioma patient derived tumor xenografts with mutational alterations that mimic those observed in patient tumors which we established can be used for preclinical development of novel drug regimens and for studying the functional aspects of BAP1 biology in mesothelioma. Electronic supplementary material The online version of this article (doi:10.1186/s12885-015-1362-2) contains supplementary material, which is available to authorized users.

Viable neurons in brain slices are crucial for electrophysiological studies. The present study describes a new method for obtaining viable cells in several regions of the central nervous system including the ventral tegmental area, the hypothalamus, the periaqueductal grey matter and the spinal cord. The essence of the method was to use a modified artificial cerebrospinal fluid (ACSF) in which all NaCl was replaced initially by equi-osmotic glycerol. This modified glycerol-based ACSF was used during slice preparation. The underlying principle for the modification is to prevent the possible acute neurotoxic effects of passive chloride entry, subsequent cell swelling and lysis. This method significantly increased the live/dead ratio in morphology compared to the normal ACSF or sucrose-base ACSF, in which NaCl was replaced by sucrose. An examination of some electrophysiological and pharmacological properties of the neurons in these preparations, by means of current-clamp and voltage-clamp recordings, revealed similar properties of those neurons obtained with the traditional ACSF method. Due to the increase in the number of viable neurons, the new ACSF increases the productivity of experiments. Based on our data, we propose that this glycerol-based solution may protect CNS neurons.

Several polyphenolic compounds, including flavonoids and phenolic acids, were compared with their per-methylated forms in both chemical and cell-based assays for antioxidant capacity. Methylation largely eliminated “chemical” antioxidant capacity, according to ferric reducing antioxidant power and oxygen radical absorbance capacity assays. Methylation, however, only moderately reduced protection of human Jurkat cells in culture, from hydrogen peroxide-mediated cytotoxicity, at physiologically relevant concentrations. Neither methylated nor un-methylated compounds were detectably metabolized by the cells. It appears that the protective mechanism of polyphenolic antioxidants against high concentrations of hydrogen peroxide in human cells may be largely unrelated to chemical antioxidant capacity.

Purpose: To determine whether mesothelin, a cell surface protein highly expressed in mesothelioma and ovarian cancer, is shed into serum and if so to accurately measure it. Experimental Design: We developed a sandwich ELISA using antibodies reacting with two different epitopes on human mesothelin. To quantitate serum mesothelin levels, a standard curve was generated using a mesothelin-Fc fusion protein. Sera from 24 healthy volunteers, 95 random hospital patients, 56 patients with mesothelioma, and 21 patients with ovarian cancer were analyzed. Serum mesothelin levels were also measured before and after surgical cytoreduction in six patients with peritoneal mesothelioma. Results: Elevated serum mesothelin levels were noted in 40 of 56 (71%) patients with mesothelioma and in 14 of 21 (67%) patients with ovarian cancer. Serum mesothelin levels were increased in 80% and 75% of the cases of mesothelioma and ovarian cancer, respectively, in which the tumors expressed mesothelin by immunohistochemistry. Out of the six patients with peritoneal mesothelioma who underwent surgery, four had elevated serum mesothelin levels before surgery. Out of these four patients, three had cytoreductive surgery and the serum mesothelin level decreased by 71% on postoperative day 1 and was undetectable by postoperative day 7. Conclusions: We developed a serum mesothelin assay that shows that mesothelin is elevated in patients with mesothelioma and ovarian cancer. The rapid decrease in mesothelin levels after surgery in patients with peritoneal mesothelioma suggests that serum mesothelin may be a useful test to monitor treatment response in mesothelin-expressing cancers.

There is growing interest both from consumers and researchers in the role that berries play in human health. The objective of this study was to investigate whether anthocyanins and other phenolics present in boysenberries and blackcurrants are effective in protecting cells against the oxidative damage induced by hydrogen peroxide (H2O2). The concentrations of polyphenols used were within the human physiological range. The data showed that SH-SY5Y human neuroblastoma cells were protected against H2O2-induced toxicity by the anthocyanins and phenolic fractions. The concurrent addition of either fractions of these berries with H2O2 significantly inhibited the increase in intracellular reactive oxygen species (ROS) production. Pre-incubation of cells with the same concentrations had no effect on the ROS level—a result that may be due to the metabolic conversion to inactive compounds. Anthocyanins and phenolic fractions of blackcurrant were better at protecting DNA of HL-60 human promyelocytic cells from damage than similar fractions from boysenberry. The phenolic extract of blackcurrant demonstrated the highest protective effect against H2O2-induced neurotoxicity, oxidative stress and DNA damage and may be a good candidate for inclusion into a processed functional food.  2006 Society of Chemical Industry

Purpose: Mesothelin is a glycosyl-phosphatidylinositol–anchored glycoprotein present on the cell surface. Mesothelin is a differentiation antigen that is highly expressed on mesothelioma, ovarian cancer, and pancreatic cancer. The existence of a spontaneous humoral immune response to mesothelin in humans has not been fully studied. Here we addressed the issue of whether mesothelin elicits a humoral immune response in patients with mesothelioma and ovarian cancer. Experimental Design: Using an ELISA, we analyzed immunoglobulin G antibodies specific for mesothelin in sera from patients with mesothelioma and epithelial ovarian cancer. Tumor specimens were examined by immunohistochemistry for mesothelin protein expression. Results: Elevated levels of mesothelin-specific antibodies were detected in the sera of 39.1% of patients with mesothelioma (27 of 69 patients) and 41.7% with epithelial ovarian cancer (10 of 24 patients) when compared with a normal control population (44 blood donors; P < 0.01 for both mesothelioma and ovarian cancer). We also found that 53% to 56% of patients with mesothelin immunostaining-positive mesothelioma and ovarian cancer had antibodies specific for mesothelin, whereas only 0% to 8% of patients with negative mesothelin immunostaining had detectable mesothelin-specific antibodies (χ2 test: P < 0.01 for mesothelioma and P = 0.025 for ovarian cancer). Conclusions: Our findings indicate that mesothelin is a new tumor antigen in patients with mesothelioma and ovarian cancer and the immunogenicity of mesothelin is associated with its high expression on the tumor cells. Mesothelin represents an excellent target for immune-based therapies.

Replication competent lentivirus (RCL) has been the major safety concern associated with applications of lentivirus-based gene transfer systems for human gene therapy. Minimization and elimination of overlaps between the packaging and the transfer vector constructs are expected to reduce the potential to generate RCL. We previously developed second- and third-generation bovine immunodeficiency virus (BIV)-based gene transfer systems. However, some sequence homologies between the vector and gag/pol packaging constructs remained. In order to minimize the sequence homologies, we recoded gag/pol with codon usage optimized for expression in human cells in this report. Expression of the recoded gag/pol was Rev/RRE independent. Thus, RRE was eliminated from the packaging construct, thereby removing a 312 bp block of homology. In addition, recoding gag/pol minimized overall homologies between the packaging and transfer vector constructs. Vectors generated by the recoded packaging construct with a four plasmid system had titers greater than 1 x 10(6) transducing units per milliliter, equivalent to those of the earlier generation systems. The vectors were functional in vitro and efficiently transduced rat pigment epithelial cells in vivo. Generation of the synthetic packaging construct provides further advances to the safety of lentiviral vectors for clinical applications.

In this study a 12 week clinical trial was conducted with Enzogenol®, a commercially available proanthocyanidin-rich flavonoid extract derived from the bark of Pinus radiata that was formulated with vitamin C. The study was to determine whether the oxidative injury markers of protein and DNA damage could be affected by the product. Twenty four (14 males and 10 females) subjects aged between 55-75 years completed the study. The group was given a twice daily dose of 240 mg of Enzogenol® and 120 mg vitamin C for 12 weeks and blood samples were collected at the start of the study before supplementation, 6 weeks and 12 weeks. Plasma samples were analysed for protein carbonyl concentrations as a measure of protein oxidation by an ELISA method. Isolated peripheral blood mononuclear cells were analysed for DNA damage using the alkaline comet assay. Protein carbonyl concentration reductions were highly significant after 6 and 12 weeks of supplementation. DNA damage reduction, as measured by the comet assay, was not significant after 6 weeks but highly significant after 12 weeks of supplementation.

Correction for ‘pH and reduction-activated polymeric prodrug nanoparticles based on a 6-thioguanine-dialdehyde sodium alginate conjugate for enhanced intracellular drug release in leukemia’ by Yanming Wan et al., Polym. Chem., 2018, DOI: 10.1039/c8py00577j.

The results in this study show that the rhodamine fluorophore can be specifically conjugated to Angiotensin II at Lys3 residue (substituted for a Val) without altering the biological activity of the parent compound. The conjugated peptide was characterized using HPLC, mass spectrometry, and N-terminal sequencing. The rhodamine-Angiotensin II binds effectively to AT1 receptor and gets internalized in clathrin coated vesicles by endocytosis. These results clearly suggest the usefulness of fluorophore-conjugated peptides in studies such as, ligand-receptor binding, and ligand-receptor complex internalization, for drug delivery using cell receptors and as an alternative to peptide hormone radioimmunoassays.

To study the clinicopathologic features, immunophenotype and prognosis of matrix-producing metaplastic carcinoma (MPC).Sixteen cases of MPC diagnosed between 2002 and 2012 in West China Hospital were identified. The clinicopathologic features were analyzed. Immunohistochemistry for E-cadherin, S-100 protein, CK5/6, HCK, PCK, CK7, CK8, p63, SMA, EMA, CD99, MSA, CK14, EGFR, ER, PR, HER2 and Ki-67 was performed with EnVision method. The clinical outcome was evaluated and compared to matched controls of invasive ductal carcinoma.All patients were women and ranged in age from 29 to 69 years (median age 48 years). The median size of primary tumor was 4 cm. Most of the tumors were well-circumscribed with expansile and multinodular appearance. Histology showed invasive carcinoma with a direct transition from carcinoma to cartilaginous/chondromyxoid matrix without an intervening spindle cell component. Tumor distribution was either nodular or diffuse. The matrix component accounted for 10%-80% of the tumor volume. All the tumors were strongly positive for S-100 protein and basal-like cytokeratin with triple negative phenotype (ER, PR and HER2 negative). Alcian blue stain was positive for the cartilaginous/chondromyxoid matrix. Compared with invasive ductal carcinoma, patients with MPC had increased locoregional recurrence (P = 0.010), increased distant recurrence (P = 0.011) and shorter disease-free survival (P = 0.017).MPC is a rare variant of mammary metaplastic carcinoma with unique characteristics of morphology and immunohistochemical staining pattern. This subtype seems to have aggressive biologic behavior.

The mesothelin (MSLN) gene encodes a 71 kilodalton (kDa) precursor protein that is processed into megakaryocytic potentiating factor (MPF), a 31 kDa protein that is secreted from the cell, and mature mesothelin (mMSLN), a 40 kDa cell surface protein. The mMSLN binds to CA125, an interaction that has been implicated in the intra-cavitary spread of mesothelioma and ovarian cancer. To better define the role of MPF and mMSLN, growth of the lung cancer cell line A549 was evaluated in immuno-deficient mice with inactivation of the Msln gene. We observed that Msln–/– mice xenografted with intraperitoneal A549 tumors survive significantly long than tumor-bearing Msln+/+ mice. When tumor-bearing Msln–/– mice are supplemented with recombinant MPF (and to a lesser extent mMSLN), most of this survival advantage is lost. These studies demonstrate that MPF and mMSLN have an important role in the growth of lung cancer cells in vivo and raise the possibility that inactivation of MPF may be a useful treatment for lung and other MSLN expressing cancers.

To explore peptide hormone-induced conformational changes, we attempted to engineer a metal-ion binding site between the cytoplasmic loops CD and EF in the angiotensin II type 1 (AT(1)) receptor. We constructed 12 double and six triple histidine mutant receptors, and tested the ability of each mutant and the wild-type to activate inositol phosphate (IP) production with and without ZnCl(2). Inhibition by ZnCl(2) in the double and triple His mutant receptors was not significant, but these mutations directly decreased the IP production. Systematic analysis of single His mutants demonstrated that the loop CD-mutants displayed 52-74% inhibition of IP production, whereas the loop EF-mutants did not affect IP production. These results indicate that the cytoplasmic loop CD-segment from Tyr(127) to Ile(130) is important for G(q/11) activation by the AT(1) receptor.

An organic-inorganic hybrid mono-Sm(III) substituted phosphotungstate KNa3[HPro]₇[Sm(α-PW₁₁O₃₉)₂]·Pro·₁₈H₂O (1) (Pro=D-proline) has been synthesized in the conventional aqueous solution and structurally characterized by elemental analyses, inductively coupled plasma atomic emission spectrometry (ICP-AES) analyses, IR spectra, UV spectra, powder X-ray diffraction (PXRD), thermogravimetric analysis (TGA) and single-crystal X-ray diffraction. The molecule of 1 consists of a classical 1:2-type [Sm(α-PW₁₁O₃₉)₂](11-) fragment and free D-proline components. It should be pointed out that the synergistic action between the in-situ formed chiral [Sm(α-PW₁₁O₃₉)₂](11-) fragment and chiral D-proline components results in the formation of the chiral 1. The luminescence emission of 1 reveals three characteristic bands that derive from the (4)G₅/₂→(6)H₅/₂, (4)G₅/₂→(6)H₇/₂ and (4)G₅/₂→(6)H₉/₂ transitions of the Sm(III) cation as well as the synergistic contribution of the O→W transitions of [α-PW₁₁O₃₉](7-) moieties and a π(*)-n or π(*)-π transitions of Pro. Its ferroelectric behavior has been measured.

The recombinant plasmid PGC was constructed for transcription unit of c-myc gene with diorientation in vitro , to make RNA probes for detection of c-myc mRNA and antisence RNA expression of tranfectant HL R 60 -9, which was obtained from HL60 cells transfected with inducible c-myc antisense RNA expression plasmid. The results from HL R 60 -9 cells induced by Cd 2+ indicated that expression of c-myc antisense RNA increased with Cd 2+ concentration and exposure time, while c-myc mRNA expression progressively reduced. Using immunohistochemical technique no c-myc P62 protein expression was detected. The incorporation of 3 H-TdR, 3 H-UR and 3 HLeu revealed significant suppression of DNA, RNA and protein biosynthesis. It is suggested that the reversion changes previously reported in malignant phenotypes of HL R 60 -9 cells and the inhibition of macromolecular biosynthesis mentioned above were associated with the blockade of c-myc gene expression by its antisense RNA.

We report a method to generate and purify large quantities of fully active mouse iNOS from E. coli, and show that calmodulin coexpression is essential to generate the active iNOS. E. coli were transformed with a plasmid containing mouse iNOS with a six-histidine tag on its N-terminus or were cotransformed with piNOS and a distinct plasmid that contained human calmodulin. Protein expression was induced by IPTG followed by culture at room temperature. Coexpression with calmodulin enabled production of active iNOS (20 mg/L culture), of which half could be recovered in pure form by sequential metal chelate and 2', 5' ADP Sepharose chromatography. The calmodulin-replete iNOS was dimeric, contained normal quantities of heme, flavins, and tightly bound calmodulin, and had high NO synthesis activity (0.7 - 1.2 mumol NO/min per mg). In contrast, calmodulin-deficient iNOS was monomeric, devoid of flavins and heme, and had no NO synthesis activity. We conclude that calmodulin is essential to fold and stabilize mouse iNOS.

Two organic-inorganic hybrid copper-lanthanide heterometallic germanotungstates KNa2H7[enH2]3[Cu(en)2(H2O)]2[Cu(en)2]2{Cu(en)2[Eu(α-GeW11O39)2]2}·13H2O (1) and Na2H4[Cu(en)2(H2O)]2[Cu(en)2]6[Cu(en)2]{Cu(en)2[La(α-GeW11O39)2]2}·12H2O (2) have been hydrothermally synthesized by reaction of K8Na2[A-α-GeW9O34]·25H2O with CuCl2·2H2O and EuCl3/LaCl3 in the presence of en (en=ethylenediamine) and structurally characterized by elemental analyses, IR spectra and single-crystal X-ray diffraction. 1 exhibits the 1D chain motif built by tetrameric {[Cu(en)2(H2O)]2[Cu(en)2]2{Cu(en)2[Eu(α-GeW11O39)2]2}}(16-) moieties through square antiprismatic K(+) cations while 2 displays the 1D architecture made by tetrameric [[Cu(en)2]6[Cu(en)2]{Cu(en)2[La(α-GeW11O39)2]2}](10-) units via octahedral [Cu(en)2](2+) cations. Furthermore, the solid-state electrochemical and electrocatalytic properties of 1 have been investigated and 1 indicates the good electrocatalytic activity for nitrite reduction. In addition, the photoluminescence property of 1 has been investigated.

Three triterpene-caffeates have been isolated from skins of a russeted apple cultivar "Merton Russet" and identified by LC-MS and NMR as betulinic acid-3-cis-caffeate, betulinic acid-3-trans-caffeate, and oleanolic acid-3-trans-caffeate. Betulinic acid-3-trans-caffeate and oleanolic acid-3-trans-caffeate were also found in russeted pear skins. These compounds have not been previously reported in apples or pears, or in any other foods. Their presence was related to suberized tissue as they were only found in russet portions of the partially russeted apple cultivar "Cox's Orange Pippin" and were not detected in the waxy apple cultivar "Royal Gala". High concentrations of betulinic acid-3-trans-caffeate were found in the bark of both "Merton Russet" and "Royal Gala" trees. The three triterpene-caffeates showed anti-inflammatory activity in vitro, inhibiting NF-κB activation with IC50's of 6-9 μM. Betulinic acid-3-trans-caffeate, the predominant compound in the apples, was immuno-modulatory at around 10 μM in the in vitro and ex vivo bioassays, boosting production of the pro-inflammatory cytokine TNFα in cells stimulated with bacterial lipopolysaccharides.

The introduction of stimuli-responsive polymers into the study of organic catalysis leads to the generation of a new kind of polymer-based stimuli-responsive recyclable catalytic system. Owing to their reversible switching properties in response to external stimuli, these systems are capable of improving the mass transports of reactants/products in aqueous solution, modulating the chemical reaction rates, and switching the catalytic process on and off. Furthermore, their stimuli-responsive properties facilitate the separation and recovery of the active catalysts from the reaction mixtures. As a fascinating approach of the controllable catalysis, these stimuli-responsive catalytic systems including thermoresponsive, pH-responsive, chemo-mechano-chemical, ionic strength-responsive, and dual-responsive, are reviewed in terms of their nanoreactors and mechanisms.

Compounds (IV), (VI), and (VIII) are characterized by powder and single crystal XRD, IR and optical diffuse reflectance spectroscopy, magnetic measurements, and luminescence spectroscopy.

In the brain, the four subunits of the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor, glutamate A1 (GluA1), GluA2, GluA3, and GluA4 form functionally different tetramers. Of these, GluA1 is very important. It forms calcium-permeable (without GluA2) AMPA receptors and induces the trafficking and integration of AMPA receptors within synaptic membranes. Increased GluA1 expression and their phosphorylation are common mechanisms for the treatment of Alzheimer's disease, schizophrenia, depression, and chronic drug addiction. Moreover, GluA1 is also involved in pain and epilepsy. Increased phosphorylation of serine831 in the GluA1 receptor is a mechanism necessary to alleviate Alzheimer's disease and depression. GluA1-/- knockout mice are used as a model of schizophrenia. A decrease in the total cell AMPA receptor currents and phosphorylation of serine845 of GluA1 is observed in chronic drug addiction. Increased expression of GluA1 causes pain and is involved in epilepsy. GluA1-promoting AMPA receptor potentiators could be used to treat Alzheimer's disease and memory loss. In conclusion, GluA1 agonists or antagonists might be effective in various disorders and conditions of the central nervous system that are based on GluA1 status at the synaptic region.

Background Many theoretical researches predicted that the larch species would decrease drastically in China under future climatic changes. However, responses of the structural and compositional changes of Gmelin larch (Larix gmelinii var. gmelinii) forests to climatic changes have rarely been reported. Methodology/Principal Findings Field survey was conducted to examine the structures and compositions of natural Gmelin larch forests along a climatic gradient. Stepwise linear regression analyses incorporating linear and quadratic components of climatic and non-climatic factors were performed on the structural and compositional attributes of those natural Gmelin larch forests. Isothermality, Max Temperature of Warmest Month (TempWarmestMonth), Precipitation of Wettest Month (PrecipWettestMonth), Precipitation Seasonality (PrecipSeasonality) and Precipitation of Driest Quarter (PrecipDriestQuarter) were observed to be effective climatic factors in controlling structure and composition of Gmelin larch forests. Isothermality significantly affected total basal area of larch, while TempWarmestMonth, PrecipWettestMonth and PrecipSeasonality significantly affected total basal area of Mongolian pine, and PrecipDriestQuarter significantly affected mean DBH of larch, stand density of larch and total basal area of spruce and fir. Conclusions/Significance The summer and winter temperatures and precipitations are all predicted to increase in future in Northeast China. Our results showed the increase of total basal area of spruce and fir, the suppression of regeneration and the decrease of stand density of larch under increased winter precipitation, and the decrease of total basal area of larch under increased summer temperature in the region of current Gmelin larch forest. Therefore, we suggest that larch would decrease and spruce and fir would increase in the region of future Gmelin larch forest.

The role of myeloid derived suppressor cells (MDSCs) in mediating tumor immunosuppression in patients with malignant mesothelioma has not been well characterized. The goal of our study was to analyze for the presence of both monocytic (Mo) and granulocytic (Gr) MDSCs in blood of mesothelioma patients, and characterize their mechanism of suppression of T cells responses. We evaluated the peripheral blood of patients (n = 23) for the presence of Gr-MDSC (CD11b+CD15+CD14-HLADR-) and Mo-MDSC (CD11b+CD14+HLADR-). Gr-MDSC (62.3±13.9% vs. 47.8±17.8%; p = 0.005) as well as monocytic MDSC (0.2±0.4% vs. 0.1±0.3%; p = 0.01) were significantly elevated in peripheral blood of mesothelioma patients as compared to healthy donors (n = 21). The frequency of Gr-MDSC was higher than Mo-MDSC. Thus, Gr-MDSCs appeared to be the main immunosuppressive population and were investigated further.Gr-MDSC from mesothelioma patients were found to inhibit the proliferation of both autologous CD4 and CD8 T cells (mean inhibition of proliferation of 61.9% for CD4 and 75.5% for CD8 at 1:2 T: Gr-MDSC ratio) and was accompanied by a decrease in IFN-γ released by T cells in the co-culture supernatants (92.4 pg/mL at 1:2 ratio). To determine the mechanism by which Gr-MDSC inhibit T cells, we tested the levels of reactive oxygen species (ROS), and nitrite oxide species and arginase activity in Gr-MDSC sorted from peripheral blood. Both arginase (0.3±0.3 vs. 0.2±0.3 μM) and nitrite levels (1.8±2.5 vs. 0 μM) in Gr-MDSC were below detection limit in both patients and donors and were marginally affected by the addition of their respective inhibitors (nor-NOHA and L-NMMA). However, the levels of ROS in Gr-MDSCs derived from mesothelioma patients were on average 3 folds higher than the Gr-myeloid cells from healthy donors (average MFI ∼15000 vs. ∼5000). In summary our results show that the major immunosuppressive cell population in mesothelioma patients are Gr-MDSCs and they suppress T cell proliferation and activation through release of reactive oxygen species. Citation Format: Swati Khanna, Francis Mussai, Anish Thomas, Gary Middleton, Constance Yuan, Betsy Morrow, Jingli Zhang, Ira Pastan, Maryalice Stetler-Stevenson, Raffit Hassan. Granulocytic MDSCs (CD11b+CD15+CD14-HLADR-) in peripheral blood of mesothelioma patients are elevated and suppress T cell proliferation and function via reactive oxygen species. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3247.

Mesothelin is a tumor differentiation antigen that is highly expressed in many cancers including malignant mesothelioma. RG7787 is a second-generation anti-mesothelin immunotoxin that is designed to be less immunogenic in patients and has reduced non-specific toxicity in pre-clinical studies. In this study, we evaluated the anti-tumor efficacy of RG7787 alone and in combination with nab-paclitaxel using primary mesothelioma cell lines obtained from patients with pleural or peritoneal mesothelioma. Four early passage malignant mesothelioma cell lines: NCI-Meso16, NCI-Meso19, NCI-Meso21 and NCI-Meso29 were evaluated for mesothelin expression and have 18×103- 56×103 mesothelin sites/cell. All four cell lines were sensitive to RG7787 with IC50s of 0.3 to 10 ng/ml. Three of the cell lines: NCI-Meso 16, NCI-Meso 21 and NCI-Meso 29 were also sensitive to nab-paclitaxel with IC50s of 4, 40 and 2 ng/ml respectively while, NCI-Meso 19 was resistant with IC50 >100 ng/ml. Synergistic in vitro cell cytotoxicity was observed with both NCI-Meso16 and NCI-Meso21 cells, when treated with the combination of RG7787 and nab-paclitaxel. To evaluate the efficacy of RG7787 with nab-paclitaxel in vivo, athymic nude mice were inoculated subcutaneously with 5×106 NCI-Meso 16 cells. When the tumors reached 100 mm3 in size, mice (n = 7 in each group) were treated with 2 cycles (separated by 3 days) of either RG7787 2.5 mg/kg every other day x 3 doses, nab-paclitaxel 100 mg/kg on day 1, a combination of RG7787 and nab-paclitaxel at the same dose and schedule as for single agent therapy, or received no treatment (control). Although treatment with RG7787 and nab-paclitaxel alone resulted in tumor shrinkage compared to control mice, none of the mice had complete tumor regressions. However, in mice treated with RG7787 plus nab-paclitaxel all mice had complete tumor regressions by day 64 that persisted in 7/7 mice when the experiment was terminated 100 days after starting treatment. In the slow growing NCI-Meso 21 in-vivo model, we also observed similar synergistic anti-tumor effects with the combination of RG7787 and nab-paclitaxel. Taken together, our findings show that RG7787 in combination with nab-paclitaxel has significant anti-tumor activity against primary mesothelioma cell lines both in vitro and in vivo, suggesting that this combination treatment could be useful to treat patients with mesothelioma. Citation Format: Jingli Zhang, Qun Jiang, Christine Alewine, Swati Khanna, Betsy Morrow, Anish Thomas, Ira Pastan, Raffit Hassan. The reduced immunogenicity anti-mesothelin immunotoxin RG7787 in combination with nab-paclitaxel has significant anti-tumor efficacy against primary mesothelioma cell lines and patient derived mesothelioma tumor xenografts. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3103.

SS1P is a recombinant immunotoxin (RIT) engineered for the targeted elimination of malignant cells that express the tumor-associated antigen mesothelin. It is composed of an antimesothelin antibody variable fragment (Fv) linked to a cytotoxic fragment of Pseudomonas exotoxin A (PE) that includes domains II and III of native PE. The clinical use of SS1P is limited by its propensity to induce neutralizing antibodies and to cause a dose-limiting capillary leak syndrome (CLS) in patients. In this article, we describe a reengineered SS1P with improved properties that overcome these deficits. The redesign of SS1P consists of (i) removing the bulk of PE domain II (residues 251–273 and 284–394 of native PE), leaving only an 11-residue furin cleavage site, (ii) adding a Gly–Gly–Ser peptide linker after the furin cleavage site, and (iii) replacing eight highly solvent-exposed residues in the catalytic domain of PE. The new molecule, SS1-LR/GGS/8M, has cytotoxic activity comparable with SS1P on several mesothelin-expressing cell lines and remarkably improved activity on primary cells from patients with mesothelioma. In a mouse xenograft tumor model, high doses of SS1-LR/GGS/8M elicit antitumor activity superior to the activity of SS1P at its maximum-tolerated dose. In addition, SS1-LR/GGS/8M has greatly decreased ability to cause CLS in a rat model and reduced antigenicity or reactivity with antibodies to the sera of patients previously treated with SS1P. Mol Cancer Ther; 12(1); 48–57. ©2012 AACR.

An efficient L1 /Cu(II)-catalyzed highly chemoselective and enantioselective nitroaldol reaction of α- or β-haloenals with nitromethane has been described. The reaction delivered chiral functionalized allylic alcohols bearing halo and nitro groups with excellent results (up to 94% yield and 99% ee) under mild reaction conditions.

We have simulated the hybrid current using Neurons in Action software to observe the hypothetical effect of Mitragyna speciosa in the model rat hippocampus CA3 pyramidal neuron at different Calcium ion conductance and at different temperature. Previous studies using the patch clamp technique showed that Mitragyna speciosa or Ketum could block long lasting Calcium channel (L-type Ca channel) currents in N1E-115 neuroblastoma cells. There is currently no such study of the effects of ketum on pyramidal neurons using the patch clamp technique. Our team is interested to study the ability of Ketum alakaloid to block L-type Ca channel in a rat's brain. The results of the simulation study gave us valuable information and guidelines for our future study to replicate the Ca channel blocking nature of the alkaloid of the Mitragyna speciosa.

Two milk-based beverages delivering twice the average daily antioxidant intake were formulated, based on synergistic combinations of fruit and vegetable extracts, and containing vitamin C (1.00 mg/ml) for shelf stability. Smokers (n = 42) consumed prototype milk A, B or non-supplemented milk (no extracts or vitamin C; 200 ml) twice daily for 6 weeks. Fasting and post-prandial (2 h after milk consumption) blood samples were collected at baseline and the end of each treatment. Non-supplemented milk significantly reduced fasting inflammatory cytokines (interleukin (IL) 6, IL-1β, tumour necrosis factor-α) compared to baseline. Both supplemented milk-based beverages significantly increased fasting plasma vitamin C concentrations and antioxidant potential and decreased serum uric acid, compared to non-supplemented milk. The beverages did not induce post-prandial oxidative stress or inflammation. Therefore, regular consumption of the supplemented milks may confer health benefits because of increased antioxidant potential or through mechanisms resulting from increased vitamin C or decreased uric acid concentrations.

The Pd/Ti electrode was prepared by coating with Pd on the surface of a Ti electrode at stable electric potential, and the reduction of pentachlorophenol (PCP) dechlorination was studied in an electro-biological system composed of the Pd/Ti electrode and the anaerobic microbes. The results showed that when the potential in the electro-biological system was −1.3V, the current density increased to the largest value which was 0.0014 A, and the efficiency of the electron transfer was higher. The efficiency of the PCP dechlorination in the electro-biological system was greater than that in single biological system by 23.3%, and the dechlorination of microbial respiration is the internal impetus during the dechlorination of PCP in the electro-microbe system. Electron transport chain of microbial respiration coupled with the hydrogen supply of the electric field, which resulted in the interaction of the electric field and biological enzyme, and the electric field in the anaerobic microbial degradation accelerated the reductive dechlorination of the PCP.

The effects of the conditions of nitrogen- and phosphorus-deficiency on the filamentous abundance index (FI), the removal efficiency of pollutants and the membrane fouling were investigated. The results indicated that the removal efficiency of CODCr reduced obviously at nitrogen deficiency and FI was higher than that at phosphorus deficiency. When filamentous bulking happened, FI increased and the micro-animals: swimming ciliates→vorticella→rotifers→Epistylis→arcella became the dominant population in turn, which could be indicative for the operation of the MBR. When FI increased from 2 to 4, Zeta potential lowered and extracellular polymeric substances (EPS) and the relative hydrophobicity (RH) in the mixed liquor rose sharply, that accelerated the membrane fouling. Filamentous bulking has the important influence on the RH of membrane, the dynamic contact angles of polluted membrane were 64°(FI=1), 67°(FI=2), 71°(FI=3), 76°(FI=4)and 81°(FI=5), respectively. The increase of EPS and sludge volume index (SVI) has reinforced the adsorption of filamentous bacteria on the membrane surface, which caused the membrane fouling to become more serious. Filamentous bulking, which influenced the operation of MBR, should be avoided in MBR.

Kiwifruit are nutrient-dense fruit with a reputation for promoting good health. Although this could be attributed to the high vitamin C content of kiwifruit, other phytochemicals could also provide health benefits. Kiwifruit are commonly reported to be a good source of vitamin E and in addition contain phenolics and carotenoids. The antioxidant properties of kiwifruit have received attention as possible mechanisms for their health-promoting effects. In this review, the antioxidant capacity of kiwifruit is discussed in the context of biologically relevant in vitro assays for predicting antioxidant activity in a biological setting compared with chemical antioxidant assays, and the ability of kiwifruit to protect cells from dying after exposure to an oxidative insult by hydrogen peroxide (cytoprotection). Some recent data are included, where extracts from twenty kiwifruit genotypes, derived from germplasm held at The New Zealand Institute for PlantFood Research Ltd, were compared for their cellular antioxidant activity and cytoprotection, using human gut-derived epithelial cell lines. Our knowledge of how this type of result is currently reflected in vivo is summarised, together with the 'naturally protective' properties of kiwifruit that involve modulating immune responses in a positive way. Finally, the ways in which these antioxidant and natural protective properties of kiwifruit may influence human health and wellness are discussed.

High performance liquid chromatography (HPLC)-based system was utilized for generating chemical fingerprints of 11 Ganoderma strains. The data were statistically evaluated by using chemometric methods in order to classify the samples. The similarities of all the 11 samples and the relative peak areas of 13 common peaks were newly calculated separately. Then different chemometrics methods including hierarchical cluster analysis (HCA), principal component analysis (PCA) and discriminate analysis (DA) were applied to classify the G. lucidum samples. Consistent results show that the application of HPLC fingerprint coupled with powerful chemometrics analysis in the discrimination and classification of Ganoderma is a reliable and scientific approach.

Flowering progress, floral morphology and nectar characteristics of Cerasus cerasoides were investigated from November 2011 to January 2012 in Kunming, Yunnan, China. We found that nectar removal had a positive effect on nectar production. The phenomenon of nectar reabsorption in C. cerasoides occurred at the end of the floral lifespan, on the tenth day after anthesis when allowing for nectar accumulation. A flower reabsorbs nectar when it has not been foraged, with the reabsorbed sugar content accounting for about 34% of the maximum sugar content. Therefore, nectar reabsorption can only reclaim a part of the energy allocated for nectar production.

Novel therapeutic agents that are safe and effective are needed for the treatment of pancreatic, ovarian, lung adenocarcinomas and mesotheliomas. Mesothelin is a glycosyl-phosphatidyl inositol (GPI)-linked membrane protein of 40 kDa over-expressed in all pancreatic adenocarcinoma and mesothelioma, in70% of ovarian adenocarcinoma, and in non-small cell lung and colorectal cancers. The biological functions of mesothelin are not known, although it appears to be involved in cell adhesion via its interaction with MUC16. We have recently developed MORAb-009, a mouse-human chimeric IgG1kappa monoclonal antibody with an affinity of 1.5 nM for human mesothelin. Here we provide evidence that MORAb-009 prevents adhesion of mesothelin-bearing tumor cells to MUC16 positive cells and can elicit cell-mediated cytotoxicity on mesothelin-bearing tumor cells. Treatment that included MORAb-009 in combination with chemotherapy led to a marked reduction in tumor growth of mesothelin-expressing tumors in nude mice compared to chemotherapy or MORAb-009 treatment alone. No adverse effects of MORAb-009 were noted during toxicology studies conducted in non-human primates. The preclinical data obtained from our studies warrants pursuing clinical testing of MORAb-009. We have in fact initiated a Phase I clinical study enrolling patients with mesothelin-positive pancreatic, mesothelioma, non-small cell lung and ovarian cancers.

This study modeled, in vitro, the potential effect of conjugative (phase II) metabolism on the cytoprotective capacity of fruit flavonoids against oxidative stress. Flavonoid aglycones were compared with their corresponding isomeric mixtures of glucuronides for their ability to enhance the survival of cultured human Jurkat T and neuroblastoma cells stressed with hydrogen peroxide. Various polyphenolic compounds were tested as substrates in vitro for an ovine liver glucuronyl transferase preparation. Flavonoids and their glycoside derivatives were found to be good substrates, whereas phenolic acids were either poor or nonsubstrates. Five common flavonoids were glucuronidated to prepare mixtures for bioassay testing. Glucuronidation generally weakened the cytoprotective capacities of flavonoids (in the presence of H(2)O(2)), but some compounds were weakened much more than others. The concentration that halved cell death was well below 0.5 microM for most flavonoids tested, but glucuronidation increased median effective concentration values to a range of 1-16 microM. This compares with the generally accepted physiological range (0.1-10 microM) for circulating dietary polyphenolics detected in the body. Therefore, some flavonoids may retain a reduced cytoprotective capacity in vitro, after glucuronidation, whereas others may be effectively inactivated.

The free radical scavenging activities of three flavonoids (quercetin, rutin and catechin) and four hydroxycinnamic acids (caffeic, ferulic, sinapic, and chlorogenic acids) were evaluated using both oxygen radical absorbance capacity (ORAC) and lipid peroxidation inhibition capacity (LPIC) assays. The cytoprotective effects of these compounds were also measured by the degree of protection against H(2)O(2)-induced damage of human Jurkat cells. All compounds exhibited protection against H(2)O(2)-mediated cytotoxicity in a dose-dependent manner. The concentrations required to result in a 50% reduction in cell death (EC(50) value) were calculated from their dose-response curves. These ranged from 0.15-2.65 microM. Overall, the four hydroxycinnamic acids tested were less effective than the three flavonoids, and of all compounds tested, quercetin offered the strongest protection against H(2)O(2)-induced cell death. A comparison of the results showed that the ability to inhibit peroxidation of lipids in a liposomal system (LPIC) correlated well with the cytoprotective activities (EC(50)), but not with the ability to protect an aqueous fluorescent substrate in the ORAC assays. The results suggest that the behavior of antioxidants in a liposomal membrane is to some extent similar to the mechanism involved in the protection of living cells from oxidative damage.