The phenolic constituents and free radical scavenging activities of Gynura formosana Kiamnra

Gynura formosana Kiamnra (Compositae) is a herbal folk medicine that is a popular vegetable in Taiwan. The free‐radical scavenging activities of a 70% aqueous acetone extract from the herb G formosana were evaluated. Bioassay‐guided fractionation, column separation on Diaion, Toyopearl HW 40(C), Sephadex LH‐20 and MCI CHP20P, and high‐performance liquid chromatography (HPLC) were used to isolate for the first time in G formosana four potent phenolics [caffeic acid ( I ), quercetin 3‐O‐rutinoside ( II ), kaempferol 3‐O‐rutinoside ( III ) and kaempferol 3‐O‐robinobioside ( IV )]. The IC₅₀ values of 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical scavenging activity for compounds I – IV were 6.7, 7.7, 300.3 and 286.7 µM , respectively, and, for superoxide radical scavenging activity, they were 187.3, 25.8, 55.3 and 87.4 µM , respectively. Using a spin trapping electron spin resonance (ESR) method, caffeic acid ( I ) and quercetin 3‐O‐rutinoside ( II ) exhibited good hydroxyl radical activity. The free radical scavenging activity of G formosana phenolics may improve the economic value of this herb and assist in its development as a health food. Copyright © 2004 Society of Chemical Industry     

The development of a suitable manufacturing process for ‘Benifuuki’ green tea beverage with anti‐allergic effects

Epigallocatechin‐3‐O‐(3‐O‐methyl) gallate (EGCG3″Me) has been reported to inhibit type I allergy better than epigallocatechin gallate (EGCG), a major catechin in tea leaves (Camellia sinensis L). We examined the effects of extraction and sterilization on the catechin content and histamine release from mast cells, as a representative reaction of early phase allergy, in the manufacture of ‘Benifuuki’ green tea beverage. Among various varieties of tea, the cultivar ‘Benifuuki’ contains approximately 2% of EGCG3″Me. Ester‐type catechins and their epimers increased with the increased extraction temperature of the tea. A tea infusion, extracted at 90 °C, strongly inhibited histamine release from mast cells. Furthermore, sterilization affected the catechin content in the manufactured green tea beverage. Sterilization at high temperature promoted the isomerization of catechins and the sterilized green tea beverage had a strong inhibitory effect. When EGCG3″Me, EGCG, epicatechin‐3‐O‐gallate (ECG) and their epimers, GCG3″Me (gallocatechin‐3‐O‐(3‐O‐methyl) gallate), GCG (gallocatechin‐3‐O‐gallate) and CG (catechin‐3‐O‐gallate) were compared, the anti‐allergic effect of GCG3″Me was strongest, and the order of activity was GCG3″Me > EGCG3″Me > GCG > EGCG. We consequently suggest that it was necessary to extract components from tea at the highest temperature possible, and to pasteurize under retort conditions (118.1 °C, 20 min), to manufacture functional green tea beverage with an anti‐allergic action. Copyright © 2005 Society of Chemical Industry     

Fate of 14C-acrylamide in roasted and ground coffee during storage

Acrylamide (AA) is formed during heating of carbohydrate rich foods in the course of the Maillard reaction. AA has been classified as probably carcinogenic to humans. Storage experiments with roasted coffee have shown that AA levels decrease depending on storage time and temperature. In the present study the fate of AA lost during storage of roasted and ground (R&G) coffee was studied, using 14C-labeled AA as radiotracer. Radiolabel was measured in coffee brew, filter residue, and volatiles. In the brew, total (14)C-label decreased during storage of R&G coffee, while activity in the filter residue built up concomitantly. [2,3-14C]-AA (14C-AA) was the only 14C-related water extractable low molecular compound in the brew detected by radio-HPLC. No formation of volatile 14C-AA-related compounds was detected during storage and coffee brewing. Close to 90% of the radiolabel in the filter residue (spent R&G coffee, spent grounds) remained firmly bound to the matrix, largely resisting extraction by aqueous ammonia, ethyl acetate, chloroform, hexane, and sequential polyenzymatic digest. Furanthiols, which are abundant as aroma components in roasted coffee, have not been found to be involved in the formation of covalent AA adducts and thus do not contribute substantially to the decrease of AA during storage.     

Dietary factors and the regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase: implications for breast cancer and development

A role for mevalonate in cancer development has long been suggested by findings that 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity is elevated in malignant cells. Increased synthesis mevalonate and mevalonate-derived nonsterol isoprenoids supports increased cell proliferation through the activation of growth-regulatory proteins and oncoproteins, and by promoting DNA synthesis. We have recently shown that mevalonate promotes the growth of human breast cancer cells both in culture and as tumors grown in nude mice. Inhibition mevalonate synthesis, therefore, may be an effective strategy to impair the growth of malignant breast cells. Several dietary compounds with known anti-cancer effects are also reported to inhibit HMG-CoA reductase activity. Here, we review evidence suggesting that inhibition of mevalonate synthesis may mediate the protective effects of cholesterol, plant isoprenoids, genistein, and long-chain n-3 polyunsaturated fatty acids (PUFAs) on experimental breast cancer.     

Extraction,identification and enzymatic synthesis of acylated derivatives of anthocyanins from jaboticaba (Myrciaria cauliflora) fruits

Polyphenols were extracted from the skin of jabuticaba fruits (Myrciaria cauliflora). Their total concentration and in vitro antioxidant activity were analysed by the DPPH and ABTS methods. The corresponding results (dry basis) were 1290 mg gallic acid equivalent (GAE)?(100 g)^?1, 98% of DPPH radical inhibition and 120 μm TEAC?g^?1 (ABTS method). All these values are at least as higher as average values reported in the literature for other fruits. A more specific analysis of the fractions of phenolic compounds was also performed by HPLC‐MS. Ellagic acid, quercetin, rutin, delphinidin‐3‐glucoside and cyanidin‐3‐glucoside were the main compounds detected; the latter two were the most abundant. The crude extract was subjected to enzymatic acylation assays in order to synthesise new esters with new potential techno‐functionalities. Palmitic acid was used as acyl donor and lipase B of Candida antactica (CALB) as biocatalyst. HPLC‐MS evidenced the formation of palmitic monoesters in connection with the delphinidin‐3‐glucoside and cyanidin‐3‐glucoside fractions.     

Contribution of cysteine and glutathione conjugates to the formation of the volatile thiols 3‐mercaptohexan‐1‐ol (3MH) and 3‐mercaptohexyl acetate (3MHA) during fermentation by Saccharomyces cerevisiae

Background and Aims: 3‐Mercaptohexan‐1‐ol (3MH) and its ester 3‐mercaptohexyl acetate (3MHA) are potent aromatic thiols that substantially contribute to varietal wine aroma. During fermentation, non‐volatile 3MH conjugates are converted by yeast to volatile 3MH and 3MHA. Two types of 3MH conjugates have been identified, S‐3‐(hexan‐1‐ol)‐L‐cysteine (Cys‐3MH) and S‐3‐(hexan‐1‐ol)‐glutathione (GSH‐3MH). Yeast‐driven formation of 3MH from these precursors has been previously demonstrated, while the relationship between 3MHA and GSH‐3MH remains to be established. This paper aims to investigate yeast conversion of GSH‐3MH to 3MH and 3MHA, and to assess the relative contribution of each individual conjugate to the 3MH/3MHA pool of finished wines. Methods and Results: Fermentation experiments were carried out in model grape juice containing Cys‐3MH and GSH‐3MH. We found 3MH formation from GSH‐3MH to be significantly less efficient than that of Cys‐3MH. Conversely, esterification of 3MH to 3MHA was higher when 3MH was formed from GSH‐3MH. Additional in vitro assays for measuring enzyme cleavage activity suggest the involvement of a different mechanism in 3MH conversion for the two precursors. Conclusions: These results indicate that although both 3MH conjugates can be converted by yeast, the type of precursor affects the rate of formation of 3MH and 3MHA during fermentation. Significance of the Study: Management of the pool of aromatic thiols during fermentation can depend on relative proportions of different 3MH conjugates.     

Urinary 2-ethyl-3-oxohexanoic acid as major metabolite of orally administered 2-ethylhexanoic acid in human

Human metabolism of 2-ethylhexanoic acid (2-EHA), which is a known metabolite of important phthalates, was investigated using 2-EHA-contaminated food. The results of our studies reveal that the major catabolic pathway of 2-EHA in human is beta-oxidation. The dominant final urinary metabolite was identified and quantified as 3-oxo-2-ethylhexanoic acid (3-oxo-2-EHA), but only after immediate methylation of the extract from urine and prior to GC-MS analysis. Former studies without the precaution of immediate methylation had found 4-heptanone as the major metabolite, which is obviously an artifact arising from the decarboxylation of 3-oxo-2-EHA.     

Absorption of N-phenylpropenoyl-L-amino acids in healthy humans by oral administration of cocoa (Theobroma cacao)

Besides flavan-3-ols, a family of N-phenylpropenoyl-L-amino acids (NPAs) has been recently identified as polyphenol/amino acid conjugates in the seeds of Theobroma cacao as well as in a variety of herbal drugs. Stimulated by reports on their biological activity, the purpose of this study was to investigate if these amides are absorbed by healthy volunteers after administration of a cocoa drink. For the first time, 12 NPAs were quantified in human urine by means of a stable isotope dilution analysis with LC-MS/MS (MRM) detection. A maximum amount was found in the urine taken 2 h after the cocoa consumption. The highest absolute amount of NPAs excreted with the urine was found for N-[4'-hydroxy-(E)-cinnamoyl]-L-aspartic acid (5), but the highest recovery rate (57.3 and 22.8%), that means the percentage amount of ingested amides excreted with the urine, were determined for N-[4'-hydroxy-(E)-cinnamoyl]-L-glutamic acid (6) and N-[4'-hydroxy-3'-methoxy-(E)-cinnamoyl]-L-tyrosine (13). In order to gain first insights into the NPA metabolism in vivo, urine samples were analyzed by LC-MS/MS before and after beta-glucuronidase/sulfatase treatment. As independent of the enzyme treatment the same NPA amounts were found in urine, there is strong evidence that these amides are metabolized neither via their O-glucuronides nor their O-sulfates. In order to screen for caffeic acid O-glucuronides as potential NPA metabolites, urine samples were screened by means of LC-MS/MS for caffeic acid 3-O-beta-D-glucuronide and 4-O-beta-D-glucuronide. But not even trace amounts of one of these glucuronides were detectable, thus excluding them as major NPA metabolites and underlining the importance of future investigations on a potential O-methylation or reduction of the N-phenylpropenoyl moiety in NPAs.     

The N-terminal amino acid of apolipoprotein D is putatively covalently bound to 3-hydroxy-3-methyl hexanoic acid, a key odour compound in axillary sweat

Axillary sweat is odourless when freshly collected at the surface of human skin, but it contains non-odoriferous precursors, which can be transformed into odorous substances by bacteria. E-3-methyl-2-hexanoic acid (3M2H) is one of the key odorous substances, but there are two contradictory reports about its precursor form. One report states that 3M2H linked non-covalently to apolipoprotein D (apoD) is the precursor, while a second report states that 3M2H-Gln identified in human axillary sweat is the precursor. Recently, 3-hydroxy-3-methyl hexanoic acid (HMHA) and 3-methyl-3-sulfanylhexane-1-ol (3M3T) have also been identified and reported as characteristic components found in apocrine sweat. To better understand the formation of axillary odours and the structural relationships between these compounds and apoD, we characterized the linkage between odorous substances and apoD in human axillary secretions. ApoD was purified from human axillary secretions collected from 50 healthy female volunteers and was then digested by trypsin and analysed by MALDI-TOF mass spectrometry. A Mascot search showed that 8 peaks identified in the trypsin-digested samples correspond to the masses calculated for theoretically digested apoD sequences and the purified protein was assigned as a precursor of apoD [Homo sapiens]. One spectrum corresponded to the theoretical peak of HMHA linked covalently to the N-terminal fragment of apoD. In contrast, no spectrum corresponded to the theoretical peak of a 3M2H adduct or to an unmodified N-terminal fragment of apoD. These results indicate a possibility that HMHA binds covalently to the N-terminal amino acid of apoD in human axillary secretions.     

The Candida albicans 14-3-3 gene, BMH1, is essential for growth.

The 14-3-3 proteins are a family of conserved small acidic proteins that have been implicated in playing major roles in a wide variety of signalling cascades. In Saccharomyces cerevisiae, the 14-3-3 genes (BMH1 and BMH2) are essential for normal pseudohyphal induction and normal bud cell development. The Bmh proteins function in the cAMP-dependent RAS/MAPK and rapamycin-sensitive signalling cascades. Deletion of only one BMH gene demonstrates no phenotypic differences under normal growth conditions. Strains deleted of both BMH1 and BMH2 are either non-viable or demonstrate sensitivity to environmental stresses. In Schizosaccharomyces pombe, the BMH homologues (RAD24 and RAD25) are essential for cell cycle control after DNA damage and deletion of both genes renders the cell inviable. The 14-3-3 gene in Candida albicans (BMH1) was identified using a novel adherence assay and differential display RT-PCR. Unlike other yeasts, C. albicans has only one 14-3-3 gene (BMH1). It was not possible to construct double knockouts by routine methods. These results suggested that the C. albicans BMH1 gene is essential. The essentiality of C. albicans BMH1 was confirmed by a PCR disruption technique. The C. albicans bmh1 Delta/BMH1 heterozygotes exhibit growth and morphogenetic defects. Therefore, the BMH1 gene in C. albicans (Accession No. AF038154) is an excellent candidate to improve our understanding of the coordinate regulation of cell cycle and morphogenesis.     

Isolation of a free radical‐scavenging antioxidant from water spinach (Ipomoea aquatica Forsk)

Ipomoea aquatica Forsk, a green leafy vegetable that is a rich source of vitamins and amino acids with many health benefits, has been explored for the isolation and identification of its bioactive compounds. Activity‐guided repeated fractionation of a methanol extract on a silica gel column followed by an XAD column yielded a compound that exhibited antioxidant activity with an EC₅₀ value of 83 ± 1.02 µg ml^?1 reaction mixture. It also showed very strong lipid peroxidation‐inhibitory activity in a liposome model system with an EC₅₀ value of 72.2 ± 0.9 µg ml^?1. However, it showed negligible metal‐chelating activity. Based on UV, 2D nuclear magnetic resonance and gas chromatography/mass spectrometry studies, the compound was tentatively identified to be 7‐O‐β‐D ‐glucopyranosyl‐dihydroquercetin‐3‐O‐α‐D ‐glucopyranoside. This is the first report on the antioxidant properties of I aquatica leaf extracts. Copyright © 2005 Society of Chemical Industry     

3‐Chloropropane‐1,2‐diol (3‐MCPD) in Soy Sauce: A Review on the Formation,Reduction, and Detection of This Potential Carcinogen

Soy sauce, a dark‐colored seasoning, is added to enhance the sensory properties of foods. Soy sauce can be consumed as a condiment or added during the preparation of food. There are 3 types of soy sauce: fermented, acid‐hydrolyzed vegetable protein (acid‐ HVP), and mixtures of these. 3‐Chloropropane‐1,2‐diol (3‐MCPD) is a heat‐produced contaminants formed during the preparation of soy sauce and was found to be a by‐product of acid‐HVP‐produced soy sauce in 1978. 3‐MCPD has been reported to be carcinogenic, nephrotoxic, and reproductively toxic in laboratory animal testing and has been registered as a chemosterilant for rodent control. 3‐MCPD is classified as a possible carcinogenic compound, and the maximum tolerated limit in food has been established at both national and international levels. The purpose of this review is to provide an overview on the detection of 3‐MCPD in soy sauce, its toxic effects, and the potential methods to reduce its concentration, especially during the production of acid‐HVP soy sauce. The methods of quantification are also critically reviewed with a focus on efficiency, suitability, and challenges encountered in analysis.