Binding of heterocyclic amines by yeast cells and their fractions

The binding of mutagenic pyrolysis products to cells of 50 yeast strains and their cell fractions was investigated. Cells of all yeast strains effectively bound 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) and 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2). Cell walls (CW), and cell wall glucan and mannan (5 mg in each case) showed the highest binding of Trp-P-1 (50 μg ml^?1); glucan adsorbed virtually all of the Trp-P-1. Cytoplasm also showed some binding but was much less effective. Glucans also bound well with 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and 2-amino-3,8-dimethylimidazo 4,5-quinoxaline (MeIQX) much more than CW, but 2-amino-5-phenylpyridine (Phe-P-1) and 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MelQ) were not effectively bound. The quantity of IQ, MeIQ, Phe-P-1 and MeIQX bound was dependent on the strain of yeast. The mutagenic pyrolysis products bound to cells were effectively extracted by aqueous methanol, ammonia (50 g litre^?1) and alcohol, but not by water. The binding was pH dependent and inhibited by metal salts. When yeast cells were heated to 100° for 15 min, the binding of Trp-P-1 decreased by about 30% but Saccharomyces cerevisiae 50 heated to 100° did not differ much from untreated cells in its binding ability.     

The binding ability of alpha-lactalbumin and beta-lactoglobulin to mutagenic heterocyclic amines.

The binding ability of bovine milk proteins with mutagenic heterocyclic amines was investigated. Binding was determined with 2 mg of beta-lactoglobulin and 20 micrograms of heterocyclic amine in .4 ml of pH 7.4, 50 mM phosphate buffer, at 37 degrees C, in a shaker for 10 min. The unbound heterocyclic amine in protein-free ultrafiltrate was analyzed by HPLC method. The binding of alpha-lactalbumin, beta-lactoglobulin A and beta-lactoglobulin B were 90.44, 81.38, and 89.18%, respectively, with 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole; 37.85, 34.04, and 43.90%, respectively, with 3-amino-1-methyl-5H-pyrido[4,3-b]indole; and 49.11, 43.25, and 57.44%, respectively, with 2-amino-6-methyldipyrido[1,2-a:3',2'-d] imidazole. Binding of beta-lactoglobulin and alpha-lactalbumin to 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole and 3-amino-1-methyl-5H-pyrido [4,3-b]-indole was higher at pH conditions above 7.4, and binding was lost at pH less than 5.5. Maximum binding of both proteins to 2-amino-6-methyl-dipyrido[1,2-a:3',2'-d]imidazole was at pH 7.4, and binding was inhibited at pH conditions above 8.5 and less than 6.5.     

The binding ability of bovine milk caseins to mutagenic heterocyclic amines.

The binding ability of bovine milk caseins with mutagenic heterocyclic amines was investigated. Binding was determined with 2 mg of casein and 20 micrograms of heterocyclic amine in .40 ml of pH 7.4, 50 mM phosphate buffer, at 37 degrees C, in a shaker for 10 min. The unbound heterocyclic amine in protein-free ultrafiltrate was analyzed by HPLC. The binding ability of whole casein, alpha s-casein, beta-casein, and kappa-casein, respectively, was 90.08, 83.06, 90.92, and 96.70% with 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole; 85.48, 51.54, 63.62, and 82.71% with 3-amino-1-methyl-5H-pyrido[4,3-b]indole; and 87.03, 59.77, 97.04, and 88.30% with 2-amino-6-methyldipyrido[1,2-a:3',2'-d]-imidazole. Higher binding of 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole and 3-amino-1-methyl-5H-pyrido[4,3-b]indole to alpha s-casein, beta-casein, and kappa-casein was observed at pH above 7.4, and the binding was inhibited at pH below 6.5. The maximum binding of 2-amino-6-methyldipyrido[1,2-a:3',2'-d]imidazole to these caseins was at pH 6.6 and 7.4. The binding was inhibited at alkaline pH above 8.5 and acidic pH below 6.5.     

Lack of formation of heterocyclic amines in fumes from frying French fries

The formation of heterocyclic amines (HAs) in the fumes from frying French fries in soybean oil or lard was studied. A high-pressure liquid chromatography method was used to determine the various HAs in fumes. Results showed that the yields of fumes produced from soybean oil when heated alone for 2 or 4 h were higher than from lard; however, a reversed trend was found when frying French fries in soybean oil and lard. Most fumes from soybean oil and lard while frying French fries were adsorbed onto the condensation apparatus, while the other portions were adsorbed onto the wool and glass beads, which were incorporated in our experimental design for collecting the fumes. The fumes from soybean oil when heated alone were found to contain three HAs, namely, 2-amino-3-methylimidazo[4,5-f]quinoxaline (IQx), 2-amino-3-methylimidazo[4,5-f ]quinoline (IQ), and 1-methyl-9H-pyrido[4,3-b ]indole (Harman), whereas two more HAs, 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ) and 3-amino-1,4-dimethyl-5H-pyrido[4,3-b ]indole (Trp-P-1), were generated in lard. Lard was more susceptible to the formation of HAs than soybean oil when both were heated alone. No HAs were detected in the fumes from French fries fried in soybean oil and lard.     

Binding of mutagens by fractions of the cell wall skeleton of lactic acid bacteria on mutagens

The binding effect of cells and cell fractions, cell wall skeleton, cytoplasm, whole cells, and cell wall skeleton treated by lysozyme and alpha-amylase at 37 degrees C for 5 h, on Trp-P-1 (3-amino-1,4-dimethyl-[5H]pyrido [4,3-b]indole) and Trp-P-2 (3-amino-1-methyl-[5H]-pyrido[4,3-b]indole) were investigated. The cell and cell wall skeleton of Streptococcus cremoris Z-25 had greater binding activity, but cytoplasm and extract of cell wall skeleton did not bind Trp-P-1 and Trp-P-2. When the cells or cell wall skeleton were treated with lysozyme and alpha-amylase, unbound Trp-P-1 and Trp-P-2 concentrations were greater than that of the untreated control. It is possible that cell walls may be involved in the binding of mutagenic pyrolyzates to lactic acid bacteria. The cell wall skeleton of S. cremoris Z-25, Lactobacillus acidophilus IFO 13951, and Bifidobacterium bifidum IFO 14252 showed binding of Trp-P-1, 2-amino-6-methyldipyrido(1,2-a:3',2'- d)imidazole, 2-amino-5-phenylpyridine, 2-amino-3-methylimidazo(4,5-f)quinoline, 2-amino-3,4-dimethylimidazo(4,5-f) quinoline, and 2-amino-3,8-dimethylimidazo(4,5-f)quinoxaline. The cell wall skeleton of S. cremoris group and Streptococcus lactis also showed the binding activity with A N-nitrosodimethylamine. The binding of Trp-P-1 to cell walls was very high, and the binding of mutagenic pyrolyzates was variable with different bacterial species. The peptidoglycan complex and polysaccharides liberated from cell wall skeleton of S. cremoris Z-25 showed strong binding of Trp-P-2. Peptidoglycans has a binding effect of about 19.86 micrograms/mg; polysaccharides had a binding effect of 14 micrograms/mg.     

Effects of various additives on the formation of heterocyclic amines in fried fish fibre

The effects of various additives on the formation of heterocyclic amines (HAs) in fried fish fibre (Trachinooephlus myops) were studied. Fried fish fibre was prepared by boiling raw snake fish, followed by deboning, eviscerating, separating of fish meat and pressing. The fish meat was subjected to frying, during which treatment the additives, such as sugar, monosodium glutamate (MSG), antioxidants and edible oil, were added. The HAs were analyzed by high-performance liquid chromatography (HPLC) with diode-array detection. Results showed that the formation of HAs was retarded after the addition of a high level of sugar (19%), and the amount of 9H-pyrido-[4,3-b]indole (Norharman), 1-methyl-9H-pyrido-[4,3-b]indole (Harman), 2-amino-9H-pyrido[2,3-b]indole (AαC) or 2-amino-3-methyl-9H-pyrido[2,3-b]indole (MeAαC) also decreased to a minimum. The total amount of HAs rose with increasing levels of MSG, and the individual HAs, Norharman, Harman, AαC and MeAαC showed the same trend. Antioxidants, such as vitamin C, α-tocopherol and BHT (Butylated hydroxytoluene), did not show any consistent effect of concentration on HAs formation. Coconut oil contributed to the highest levels of HAs formation, followed by lard and soybean oil.     

Selective in vitro binding of dietary mutagens, individually or in combination, by lactic acid bacteria.

Specific strains of lactic acid bacteria possessing antimutagenic properties are suggested to remove mutagenic contaminants of foods through binding and an investigation of their substrate specificity is required. The ability of Lactobacillus rhamnosus strains GG and LC-705 in viable and non-viable (heat- and acid-treated) forms to remove both dietary mutagens and other aromatic dietary substrates from solution was studied using HPLC. Overall, removal increased in the order: caffeine = vitamin B12 =folic acid < ochratoxin A < aflatoxin B1 = PhIP (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine) < Trp-P-1 (3-amino-1, 4-dimethyl-5H-pyrido[4,3-b]indole) (p < 0.05). Aflatoxin B1, Trp-P-1 and PhIP were removed in high amounts (77-95%) and ochratoxin A was removed in moderate amounts (36-76%). By contrast, only minimal amounts of caffeine, vitamin B12 andfolic acid were removed (9-28%). The significant removal of selected mutagens, but not other substrates, suggests these strains may be useful for dietary detoxification. Since exposure to multiple mutagens is likely, the removal of aflatoxin B1 and Trp-P-1 from a mixture of these substrates was also investigated. Removal of AFB1 significantly increased (p < 0.05) in the presence of Trp-P-1, while removal of Trp-P-1 significantly decreased (p < 0.05) in the presence of AFB1. Overall, no significant differences in removal were found between bacterial strains or between viable, heat- and acid-treated bacteria.     

Antimutagenicity and binding of lactic acid bacteria from a Chinese cheese to mutagenic pyrolyzates

The microbiological characteristics of Nai Ge Da, a traditional cheese in China, was investigated. The lactic acid bacteria species were identified as Streptococcus cremoris, Streptococcus lactis ssp. diacetylactis, Streptococcus faecalis, and Leuconostoc paramesenteroides, Leuconostoc dextranicum, and Leuconostoc mesenteroides. Streptococcus cremoris, and Leuconostoc paramesenteroides were the dominant strains. The antimutagenicities and binding of strains of S. cremoris and S. lactic ssp. diacetylactis to mutagenic pyrolyzates was investigated. The lyophilized cells of strains showed the highest inhibitory effect on the 3-amino-1,4-dimethyl-[5H]pyrido[4,3-b] indol (Trp-P-1) and 3-amino-1-methyl-[5H]pyrido[4,3-b] indol (Trp-P-2), but many strains had no significant inhibition against 2-amino-6-methyldipyrido[1,2-a:3',2'-d] imidazole (Glu-P-1). The Trp-P-1 and Trp-P-2 were effectively bound to all bacterial cells, but binding of Glu-P-1 to cells was not effective. Among the strains tested, S. cremoris C-25 not only indicated highest binding to Trp-P-1 and Trp-P-2, but it also it had a 25.36% binding ability with Glu-P-1. When all strains were autoclaved for 15 min at 120 degrees C, binding ability to mutagens was reduced by 3 to 19%. The decrease of binding ability of S. cremoris C-25 was much less, being only 2%. After heating at 80 degrees C for 3 h, the binding ability of all strains to mutagenic pyrolyzates was not much different from those of parent viable cells.     

熟肉制品中杂环胺的形成与抑制研究进展

杂环胺是熟肉制品中最常见的危害物,其种类繁多,形成途径复杂,抑制机制也不尽相同。本文介绍了2 类杂环胺的结构,主要阐述喹啉和喹喔啉类、2-氨基-1-甲基-6-苯基咪唑[4,5-b]吡啶、β-咔啉类（1-甲基-9H-吡啶[4,3-b]吲哚（1-methyl-9H-pyrido[4,3-b]indole,harman）、9H-吡啶[4,3-b]吲哚（9H-pyrido[4,3-b]indole,norharman））以及结合态杂环胺的形成途径,归纳减少杂环胺形成的措施及机理（控制加工条件和前体物、添加外源物质清除自由基和中间体、抑制美拉德反应等）,并对目前的研究现状进行概括。     

Influence of extra virgin olive oil on the formation of heterocyclic amines in roasted beef steak

In this study, heterocyclic anime (HCA) contents were monitored in commonly consumed pan-fried beefsteak based on the highest level of human exposure. Effect of addition of extra virgin olive oil (EVOO) on HCAs formation in fried beef steaks was evaluated. After EVOO was spread on the meat surface, the raw beef was cooked at 200°C for 5 min on each side. The HCAs were extracted from the meat samples and purified using a solid-phase extraction method and then analyzed by liquid chromatography-mass spectrometry (LC-MS). Among the 15 HCAs, 3-amino-1,4-dimethyl-5H-pyrido-[4,3-b]indole (Trp-P-1), 3-amino-1-methyl-5H-pyrido [4,3-b]indole (Trp-P-2), 9H-pyrido [3,4-b]indole (Norharman), 1-methyl-9H-pyrido [3, 4-b]indole (Harman), 2-amino-9H-pyrido [2,3-b]indole (AαC), 2-amino-3-methyl-9H-pyrido [2,3-b]indole (MeAαC), 2-amino-3,8-dimethylimidazo [4,5-f]-quinoxaline (MeIQx), and 2-amino-1-methyl-6-phenylimidazo [4,5-b]-pyridine (PhIP) were detected in all of the cooked beefsteaks. HCAs formation was significantly reduced (p<0.05) when the EVOO was added to the beef prior to cooking. The addition of 2 and 4 g of EVOO considerably inhibited HCAs formation in the fried beefsteak. However, adding excess amounts of EVOO promoted some HCAs formation.     

Studies on antimutagenic effect of milk cultured with lactic acid bacteria on the Trp-P2-induced mutagenicity to TA98 strain of Salmonella typhimurium.

The inhibitory effects of cultured milk using 76 strains of lactic acid bacteria isolated from milk products were investigated on the mutagenicity of 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P2), a tryptophan pyrolysate for Salmonella typhimurium TA98. Each cultured milk sample displayed its characteristic antimutagenic effect against the mutagenicity of Trp-P2. The milk cultured with Lactobacillus acidophilus LA106 (LA2) showed the highest inhibition of 82.1% among the strains used. Milk samples cultured with Lactococcus lactis subsp. lactis, Lll103 (10-3) and Lll102 (KM) also exhibited higher inhibition percentages.     

植物多酚抑制热加工肉制品中杂环胺形成机制研究进展

杂环胺（heterocyclic amines,HAs）是肉制品热加工过程中产生的一类具有极强致畸致癌活性的有毒化合物,其中2-氨基-3-甲基咪唑并[4,5-f]喹啉（2-amino-3-methyl-imidazo[4,5-f]-quinoline,IQ）和2-氨基-3,4-二甲基咪唑并[4,5-f]喹啉（2-amino-3,4-dimethyl-imidazo[4,5-f]-quinoline,MeIQ）,2-氨基-1-甲基-6-苯基咪唑并[4,5-b]吡啶（2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine,PhIP）,1-甲基-9H-吡啶并[3,4-b]吲哚（1-methyl-9Hpyrido[3,4-b]indole,Harman）和9H-吡啶并[3,4-b]吲哚（9H-pyrido[3,4-b]indole,Norharman）3类HAs在热加工肉制品中较为常见且分布广泛。本文即围绕上述3类HAs,简要分析其形成的途径及机制,并详细阐述外源添加植物多酚抑制HAs形成的作用机制,最后依据现有研究归纳总结出多酚化合物抑制HAs与其化...     

3种熏烤鸡肉制品中杂环胺含量的检测与比较

目的 以3种熏烤鸡肉制品(鸡胗、鸡翅、鸡爪)为研究对象, 通过高效液相色谱测定其中5种杂环胺类物质(heterocyclic aromatic amines, HAAs), 即2-氨基-3-甲基咪唑并[4,5-f]喹啉(2-Amino-3- methylimidazo(4,5-f)quinoline, IQ)、2-氨基-1-甲基-6-苯基-咪唑并[4,5-b]吡啶(2-Amino-1-methyl- 6-phenylimidazo(4,5-b)pyridine, PhIP)、9H-吡啶并[4,3-b]吲哚(9H-pyrido[4,3-b]indole, Norharman)、1-甲基-9H-吡啶并[4,3-b]吲哚(1-methyl-9H-pyrido[4,3-b]indol, Harman)、2-氨基-9H-吡啶并[2,3-b]吲哚(2-amine-9H-pyrido[2,3-b]indol, AaC)的含量并进行比较。方法 采用二元流动相乙腈/醋酸-醋酸铵(pH=3.4)进行梯度洗脱, 其中乙腈/醋酸-醋酸铵随时间配比为: 0 min: 10%/90%; 10 min: 20%/80%; 25 min: 50%/50%; 35 min: 10%/90%。紫外检测波长263 nm; 荧光激发波长/发射波长随时间改变为0 min: 300/440 nm; 17 min: 315/390 nm; 21 min: 335/410 nm。结果 在3种鸡肉熏烤制品中提取的5种杂环胺类物质中, PhIP的含量最高, 且在鸡胗中最为显著, 高达152.38 ng/g, 工业化生产的鸡胗中PhIP含量仅为4.23 ng/g; 各产品中Harman的含量最低, 且在鸡爪产品及工业化生产的鸡翅产品中未检出; 熏烤的鸡肉制品中提取的IQ、PhIP、Norharman、Harman、AaC的含量均高于工业化产品。结论 工业化生产的鸡胗、鸡翅、鸡爪中的5种HAAs的含量均低于3家传统熏烤产品的含量, 传统熏烤条件下鸡肉制品的食用安全系数相对较低, 工业化产品的安全性相对较高。     

花椒叶提取物对烤牛肉饼杂环胺形成的影响

以牛肉和花椒叶为研究对象,研究花椒叶提取物（Zanthoxylum bungeanum Maxim. leaf extract,ZME）添加量（0.015%、0.030%、0.045%）对烤牛肉饼中杂环胺（heterocyclic amines,HAs）形成的影响。结果表明,与对照组相比,ZME能够显著抑制烤牛肉饼中总HAs的形成（P＜0.05）,对其最大抑制率为39.87%,但对不同种类HAs形成的影响不同。其中,添加0.045% ZME对2-氨基-1-甲基-6-苯基咪唑并[4,5-b]吡啶、2-氨基-3-甲基咪唑并[4,5-f]喹啉、2-氨基-3,4-二甲基咪唑并[4,5-f]喹啉、1-甲基-9H-吡啶并[3,4-b]吲哚和2-氨基-9H-吡啶并[2,3-b]吲哚的抑制率分别达到71.76%、78.02%、49.07%、35.82%和100%;而0.045% ZME却促进2-氨基-3,8-二甲基咪唑并[4,5-f]喹喔啉和9H-吡啶并[3,4-b]吲哚的形成。进一步分析ZME对烤牛肉饼中HAs前体物的影响,结果显示前体物的消耗随着ZME添加量的增加呈降低趋势,相关性分析表明HAs的形成与多种游离氨基酸的消耗显著相关。上述结果表明ZME能显著抑制烤牛肉饼中HAs的形成,为其提高加工食品安全性及更广泛的应用提供理论依据。     

Identification of antimutagenic activities in the extract of an edible brown alga,Hijikia fusiforme (Hijiki) by umu gene expression system in Salmonella typhimurium (TA 1535/pSK 1002)

A significant antimutagenic activity was found in the hot-water-soluble extract of an edible brown alga, Hijikia fusiforme (Hijiki in Japanese) which showed a significant inhibitory effect on the umu gene expression system in SOS response against DNA damage of Salmonella typhimurium induced by typical genotoxic substances such as N-methyl-N′-nitro-nitrosoguanidine (MNNG), furylfuramide (AF-2), 2-acetylaminofluorene(2AAF) and 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), This activity was divided into polysaccharide and non-polysaccharide fractions and the former fraction showed a relatively high antimutagenic activity compared with that of the latter fraction. The polysaccharide fraction caused roughly equal inhibitory effects on MNNG-, AF-2-, 2-AAF- or Trp-P-1-induced mutagenesis and the major portion of the antimutagenic activity of this fraction was estimated to have a molecular weight of more than 30 kDa. The non-polysaccharide fraction also exhibited significant antimutagenic activities against MNNG-, AF-2-, 2-AAF- or Trp-P-1-induced mutagenesis and the major activity or this fraction was associated with low-molecular-weight substances with less than 3.5 kDa. We discuss the biological significance of the antimutagenic activities from H fusiforme which were compared with those of Laminaria japonica and Undaria pinnatifida reported previously.     

Influence of fructooligosaccharides and garlic on formation of heterocyclic amines in fried ground beef patties

The effects of fructooligosaccharide (FOS) and garlic on the formation of 15 heterocyclic amines (HCAs) were evaluated in fried beef patties. The HCAs were extracted from the fried meat samples and purified using a solid-phase extraction method and then analyzed on a liquid chromatography-mass spectrometry. Among the 15 HCAs, 3-amino-1, 4-dimethyl-5H-pyrido-[4,3-b]indole (Trp-P-1), 3-amino-1-methyl-5H-pyrido [4,3-b]indole (Trp-P-2), 2-amino-6-methyldipyrido [1,2-a:3′,2′-d]imidazole (Glu-P-1), 2-aminodipyrido [1,2-a:3′,2′-d]imidazole (Glu-P-2), 9H-pyrido [3,4-b]indole (norharman), 1-methyl-9H-pyrido [3,4-b]indole (harman), 2-amino-9H-pyrido [2,3-b]indole (AαC), 2-amino-3,8 dimethylimidazo [4,5-f]-quinoxaline (MeIQx), and 2-amino-1-methyl-6-phenylimidazo [4,5-b]-pyridine (PhIP) were detected in all of the cooked beef patties. Analysis of variance revealed that the addition of 1 or 3 g of FOS significantly reduced the formation of total amino-carboline type HCAs in the cooked beef patties, and adding the 1 or 3 g of FOS to ground beef patties reduced levels of PhIP and MeIQx (amino-imidazo-azaarenes; AIAs) in the patties. When it is compared with the HCA formation in control, additions of minced garlic (5.0, 10.0, and 15.0 g) to the ground beef patties (100 g) reduced HCA formation in the range of 14 to 100%.     

Antimutagenicity and mutagen-binding activation of mutagenic pyrolyzates by microorganisms isolated from japanese miso

The microbiological flora of miso, a traditional fermented food in Japan, were investigated. Bacteria, a yeast and a mould were isolated and identified as Pediococcus acidilactici, Leuconostoc paramesenteroides, Micrococcus halobius, Zygosaccharomyces rouxii and Aspergillus sp. P acidilactici strains were dominant bacteria in miso. The binding and antimutagenic activities of all microbial strains towards mutagenic pyrolysates were investigated. The lyophilised cells of strains of the bacteria and yeast showed the largest antimutagenic effect on 3-amino-l-methyl-[⁵H]pyrido[4,3-b]indole (Trp-P-2), but the mould was less antimutagenic than the bacteria and yeast. Most strains tested had no effective antimutagenic activity against 2-amino-3-methyl imidazo[4,5-f] quinoline (IQ). Trp-P-2 was effectively bound by all non-mould strains but binding of IQ to cells was much less effective. Among the strains tested, Leuconostoc paramesenteroides No 28 indicated the highest binding activity, not only to Trp-P-2 but also to IQ (30% binding capacity). As the concentration of Trp-P-2 was increased, the limits of binding ability of P acidilactici No 23, Z rouxii No 6 and Aspergillus sp 1 were 650, 500 and 400μg per 5 mg respectively. The binding ability and antimutagenicity for Trp-P-2 of all strains was reduced by autoclaving at 100°C for 5 min or 121°C for 15 min, by 6–20% and 7–28%, respectively. Aspergillus sp 1 was unaffected by autoclaving. The binding ability and antimutagenicity of cell walls towards Trp-P-2 was very high, being more than 85 YO effective, but it was lower than that of cytoplasm.     

Suppressive effect of poly-gamma-glutamate on SOS response of Salmonella typhimurium induced by chemical mutagens

We studied the suppressive effect of poly-gamma-glutamate (PGA) on the SOS response of Salmonella typhimurium induced by several direct [furylframide, AF-2; N-methyl-N'-nitro-N-nitrosoguanidine, MNNG; and 4-nitroquinoline 1-oxide, 4NQO] and indirect [3-amino-1-methyl-5H-pyrido-(4,3-b) indole, Trp-P-2; 2-amino-3-methylimidazo (4,5-f) quinoline, IQ; and 2-amino-3,8-dimethylimidazo (4,5-f) quinoxaline, MeIQx] mutagens. PGA preparations with average molecular masses of 50, 2000, 4000, 6000, and 8000 kDa from Bacillus subtilis (chungkookjang) were used. When we used PGA Na salt with a molecular mass of 4000 kDa, the suppression rate increased with increasing PGA concentration; 3% PGA showed 80-90% suppression irrespective of the type of chemical mutagen. PGA preparations with molecular masses of 50 kDa and more than 6000 kDa were less effective. Glutamate and acidic polymers such as carboxymethylcellulose and xanthan gum showed lower suppressive effects than PGA. PGA proved to have high antimutagenic activity.     

Formation of heterocyclic amines in a model system during heating

The formation of heterocyclic amines (HAs) in a model system during heating at 100, 150, or 200 degrees C for varied lengths of time was studied. The following systems were used: (i) phenylalanine, (ii) creatinine, (iii) phenylalanine and glucose, (iv) phenylalanine and creatinine, (v) glucose and creatinine, and (vi) phenylalanine, glucose, and creatinine. Results showed that HAs could only be found in both systems of glucose and creatinine, and of phenylalanine, glucose, and creatinine. Both 2-amino-9H-pyrido-[2,3-b]indole (AalphaC) and 2-amino-3-methyl-9H-pyrido-[2,3-b]indole (MeAalphaC) were observed in the former system, and the formation rate of AalphaC was higher than MeAalphaC. For the latter system, 2-amino-3-dimethylimidazo[4,5-f]quinoxaline (IQx), 9H-pyrido-[4,3-b]indole (Norharman), 1-methyl-9H-pyrido-[4,3-b]indole (Harman), AalphaC, and MeAalphaC were present. The formation rate of AalphaC was the highest at 200 degrees C, followed by MeAalphaC, Harman, Norharman, and IQx. In addition, the varieties of HAs increased both with increasing heating time and temperature. The amounts of HAs formed were less in the model system of glucose and creatinine than that of phenylalanine, glucose, and creatinine. The same trend was also observed for the formation rate of HAs. At 150 or 200 degrees C, the amounts of IQx, Harman, Norharman, AalphaC, and MeAalphaC increased in the initial period of heating and then reached a plateau. The results of this study provided a basic understanding of the conditions that result in the formation of HAs during cooking of meat and fish products.     

Consumption of Brussels sprouts protects peripheral human lymphocytes against 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and oxidative DNA-damage: results of a controlled human intervention trial

To find out if the cancer protective effects of Brussels sprouts seen in epidemiological studies are due to protection against DNA-damage, an intervention trial was conducted in which the impact of vegetable consumption on DNA-stability was monitored in lymphocytes with the comet assay. After consumption of the sprouts (300 g/p/d, n = 8), a reduction of DNA-migration (97%) induced by the heterocyclic aromatic amine 2-amino-1-methyl-6-phenyl-imidazo-[4,5-b]pyridine (PhIP) was observed whereas no effect was seen with 3-amino-1-methyl-5H-pyrido[4,3-b]-indole (Trp-P-2). This effect protection may be due to inhibition of sulfotransferase 1A1, which plays a key role in the activation of PhIP. In addition, a decrease of the endogenous formation of oxidized bases was observed and DNA-damage caused by hydrogen peroxide was significantly (39%) lower after the intervention. These effects could not be explained by induction of antioxidant enzymes glutathione peroxidase and superoxide dismutase, but in vitro experiments indicate that sprouts contain compounds, which act as direct scavengers of reactive oxygen species. Serum vitamin C levels were increased by 37% after sprout consumption but no correlations were seen between prevention of DNA-damage and individual alterations of the vitamin levels. Our study shows for the first time that sprout consumption leads to inhibition of sulfotransferases in humans and to protection against PhIP and oxidative DNA-damage.