Photodegradation of 1-(2-chlorobenzoyl)-3-(4-chlorophenyl) urea in different media and toxicity of its reaction products

The photodegradation of 1-(2-chlorobenzoyl)-3-(4-chlorophenyl) urea in simulated air, methanol, dioxane, hexane, and water, with a xenon lamp as light source, was studied. The rate constants and half-lives of this compound in various media under nitrogen or oxygen were determined. The photoreaction products were analyzed with HPLC-UV, GC-MS, and direct probe MS and found to have some differences in different cases. With (32)P postlabeling DNA adduct formation experiments, one of the main products, 2-chorobenzamide, was found to be able to form a DNA adduct.     

Protection against Trp-P-2 DNA adduct formation in C57bl6 mice by purpurin is accompanied by induction of cytochrome P450

Purpurin, an anthraquinone constituent from madder root, has previously been reported as antimutagenic in the Ames Salmonella bacterial mutagenicity assay and as antigenotoxic in Drosophila melanogaster, against a range of environmental carcinogens. Short-term dietary supplementation with purpurin inhibits the formation of hepatic DNA adducts in male C57bl6 mice after a single dose of the heterocyclic amine dietary carcinogen Trp-P-2 (30 mg/kg). Inhibition of adduct formation was dose-dependent. No DNA adducts were observed in animals treated only with purpurin. The decrease in adduct formation was accompanied by significant, dose-dependent inductions of hepatic cytochrome P450-dependent dealkylations of methoxy- (CYP1A2), ethoxy- (CYP1A1), and pentoxy- (CYP2B) resorufins, total cytochrome P450, and NADPH cytochrome P450 reductase. It is hypothesized that purpurin exhibits chemopreventive potential by inhibiting the cytochrome P450-dependent metabolism of heterocyclic amines to their genotoxic N-hydroxylamines.     

Acrylamide: a dietary carcinogen formed in vivo?

Acrylamide, a chemical formed during heating of human foods, reacts with N-terminal valine in hemoglobin (Hb) and forms stable reaction products (adducts). These adducts to N-terminal valine in Hb have been used to estimate daily intake of acrylamide. Daily intake of acrylamide estimated from Hb adduct levels was higher than daily intake estimated from dietary questionnaires, possibly indicating other sources of exposures. Therefore, in this study the possible endogenous formation of acrylamide was investigated by treating mice with FeSO 4, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-hydrochloric acid (MPTP), or methamphetamine (METH). Acrylamide Hb adducts were determined, and a significant increase ( p < 0.05) in acrylamide Hb adduct levels was observed 24 h following treatment with FeSO 4 and 72 h following treatment with MPTP or METH. The results of this study show that acrylamide Hb adduct levels are increased in mice treated with compounds known to induce free radicals, thus suggesting the endogenous production of acrylamide.     

Detection and characterization of DNA adducts formed from metabolites of the fungicide ortho-phenylphenol

The significance of DNA adduction in ortho-phenylphenol-induced carcinogenesis remains unclear. Establishing adduct structures may contribute to resolving this issue. The chemical structures of the DNA adduction products resulting from the in vitro reaction of phenylbenzoquinone, the putative ultimate carcinogenic metabolite of the fungicide/disinfectant ortho-phenylphenol, are reported here. Three isomeric adducts that resulted from reaction of deoxyguanosine were characterized by UV, LC-ESI-MS, and MS/MS, and 1D and 2D COSY-NMR spectroscopy. The proposed mechanism of product formation is nucleophilic attack by the deoxyguanosine exocyclic amine nitrogen on an electrophilic quinone carbon, followed by stabilization through enolization. Another nucleophilic attack forms a five-membered ring, which aromatizes by dehydration to form the final product. Adducts were also characterized from deoxyadenosine and deoxycytidine, although conversions were at least 10 times lower. Structures are also proposed for these products. Cell culture studies confirmed that HepG2 cells incubated with phenylbenzoquinone at concentrations associated with cytotoxicity form the same DNA adducts.     

Identification of adducts between an odoriferous volatile thiol and oxidized grape phenolic compounds: kinetic study of adduct formation under chemical and enzymatic oxidation conditions

HPLC-MS and (1)H, (13)C, and 2D NMR analyses were used to identify new addition products between 3-sulfanylhexan-1-ol (3SH) and o-quinones derived from (+)-catechin, (-)-epicatechin, and caftaric acid. The kinetics of formation of these adducts were monitored in a wine model solution and in a must-like medium by HPLC-UV-MS with the aim of understanding the chemical mechanism involved in reactions between volatile thiols and o-quinones. One o-quinone-caftaric acid/3SH adduct, three o-quinone-(+)-catechin/3SH adducts, and three o-quinone-(-)-epicatechin/3SH adducts were characterized. Caftaric acid was oxidized faster than (-)-epicatechin and (+)-catechin when these phenolic compounds were incubated in a one-component mixture with polyphenoloxidase (PPO) in the presence of 3SH. Consequently, o-quinone-caftaric acid formed adducts with 3SH more rapidly than o-quinone-(+)-catechin and o-quinone-(-)-epicatechin in the absence of other nucleophilic species. Furthermore, o-quinone-(-)-epicatechin reacted faster than o-quinone-(+)-catechin with 3SH. Sulfur dioxide decreased the yield of adduct formation to a significant extent. Under chemical oxidation conditions, the rates and yields of adduct formation were lower than those observed in the presence of PPO, and o-quinone-caftaric acid was slightly less reactive with 3SH, compared to oxidized flavan-3-ols. The identification of o-quinone-caftaric acid/3SH and o-quinone-(+)-catechin/3SH adducts in a must matrix suggests that the proposed reaction mechanism is responsible for 3SH loss in dry wines during their vinification and aging process.     

Inhibition of acrylamide toxicity in mice by three dietary constituents

The inhibitory effects of three dietary constituents, tea polyphenols, resveratrol, and diallyl trisulfide, on acrylamide-biomacromolecule (liver DNA, protamine, and hemoglobin) adduct formation at human exposure level were studied by accelerator mass spectrometry. The results demonstrated that the three dietary constituents all significantly inhibited the formation of acrylamide adducts with liver DNA, whereas tea polyphenols and diallyl trisulfide reduced protamine and hemoglobin adducts, respectively. Further biochemical studies showed that acrylamide could significantly inactivate creatine kinase and glutathione S-transferase and deplete glutathione. When the inhibitors were cotreated with acrylamide, all of them could effectively recover the activities of creatine kinase. In addition, tea polyphenols and diallyl trisulfide could increase glutathione S-transferase activity remarkably. On the basis of these results, mechanisms of the effects are discussed. This study might provide a beneficial guide to people's diet for the purpose of reducing the harmful effect of acrylamide.     

蚯蚓分子生物标志物在土壤氯代苯胺类污染评价中的应用研究

采用紫外分光光度法测定DNA加合物的形成及Western-Blot技术检测蚯蚓热休克蛋白70（HSP70）的表达,研究了蚯蚓DNA加合物和HSP70作为分子生物标志物在土壤氯代苯胺化合物污染评价中的应用。DNA加合实验的结果表明,随着3,4-二氯苯胺浓度的增加（0.77、1.55、3.10 mg.mL-1）,特征吸收峰的位移发生显著改变,交联率呈现负相关;Western-Blot检测结果表明,3,4-二氯苯胺处理后蚯蚓HSP70表达增加,与空白组比较有显著性差异。     

Inhibitory effect(s) of polymeric black tea polyphenol fractions on the formation of [(3)H]-B(a)P-derived DNA adducts

Five polymeric black tea polyphenol fractions (PBP-1-5) were isolated from a popular brand of black tea. The effect of these PBPs and epigallocatechin gallate (EGCG), a major green tea polyphenol, was studied on the formation of [(3)H]-B(a)P-derived DNA adducts in vitro, employing rat liver microsomes. PBP-1-3 inhibited microsome-catalyzed [(3)H]-B(a)P-derived DNA adduct formation in vitro in a dose-dependent manner. This inhibition was further enhanced on preincubation of microsomes with each of the PBPs. PBP-4 was not effective per se and required preincubation with microsomes to exhibit its inhibitory effect, whereas PBP-5 remained ineffective with or without preincubation with microsomes. Further investigations revealed that the observed decrease in [(3)H]-B(a)P-DNA adduct formation was due to inhibition of isozymes of CYP450s by PBPs. Overall, results suggest that polymeric black tea polyphenol fractions retain one of the chemopreventive effects exhibited by the monomeric green tea polyphenol EGCG in vitro.     

不同啶虫脒剂型对烟粉虱的毒力差异及原因分析

为探讨农药剂型对烟粉虱的毒力差异,分别以甘蓝和黄瓜叶片作为生物测定载体,比较了啶虫脒乳油(EC)、微乳剂(ME)、可溶性液剂(SL)、可溶性粉剂(SP)及可湿性粉剂(WP)等5种剂型对烟粉虱成虫的毒力,并通过对植物叶片临界表面张力、啶虫脒药液表面张力、动态接触角及叶片持液量的测定,分析了不同啶虫脒剂型间毒力差异形成的原因。结果表明:以甘蓝叶片为生物测定载体,不同啶虫脒剂型间对烟粉虱的毒力差异明显;以黄瓜叶片为生物测定载体,不同剂型间毒力差异不明显。啶虫脒SL、SP、WP在两种生物测定载体间的毒力差异大于EC和ME。甘蓝和黄瓜叶片的临界表面张力值分别为30.73 mN.m 1和57.91~63.30 mN.m 1。啶虫脒有效成分浓度大于7.81 mg.L 1时,啶虫脒EC和ME溶液表面张力即小于甘蓝和黄瓜叶片的临界表面张力;啶虫脒有效成分浓度大于500 mg.L 1时,啶虫脒SL、SP、WP溶液的表面张力才小于甘蓝叶片的临界表面张力。啶虫脒ME溶液液滴接触两种植物叶片的瞬间(0 s),液滴与叶片间的接触角就明显小于90°。甘蓝叶片对低浓度的啶虫脒ME和EC溶液的持液量高于其他3种剂型,黄瓜叶片对啶虫脒各剂型溶液的持液量无明显差异。植物叶片表面性质和不同剂型溶液表面张力的差异是导致剂型间产生毒力变化的重要原因。     

Dynamics of pesticides in potato crops

This study investigated the behavior of pesticides commonly used on potato in a part of northwestern Spain with a large area devoted to this crop. Experimental potato plots were treated with commercial formulations of two insecticides, a nematicide, a herbicide, and a combination of two fungicides, and the concentrations of the active ingredients were monitored throughout the growing season in the 0-1 and 1-15 cm soil layers, in water dripping off the potato plants, and in the potatoes themselves. The technique used for pesticide determination was gas chromatography with mass selective detection. For potato analyses, pesticide extraction was optimized. The only pesticide ever detected in potato tubers was metalaxyl, the concentration of which never exceeded half the maximum residual limit even when it was applied several times more often than is officially recommended for potato crops. Metalaxyl was also the only pesticide detected in wash-off, apparently due to its being more soluble in water than the other pesticides applied to the growing plants. A mathematical model of pesticide transport in soil was fitted to the experimental data using the program HYDRUS-1D.     

Performance characteristics of methods of analysis used for regulatory purposes. Part II. Pesticide formulations.

The precision parameters of the method-performance (collaborative) studies published in the AOAC Journal from 1915 through 1990 for pesticide formulations have been recalculated on a uniform basis by the International Union of Pure and Applied Chemistry 1987 protocol. About 93% of the 953 accepted assays, which are predominantly gravimetric (G), volumetric (V), and gas (GC) and liquid (LC) chromatographic methods, exhibit relative standard deviations among laboratories (RSDR) that are generally less than 2 times the values predicted from the Horwitz equation: RSDR (%) = 2 exp (1-0.5 log C), where C is the concentration expressed as a decimal fraction. UV, VIS, and IR spectrophotometric (S) methods are somewhat poorer, with about 80% of the reported RSDR values less than twice the predicted RSDR value. The precision parameters of pesticide formulations analyzed by the older methods (G, V, GC) are equivalent to those previously found for drug preparations in the same concentration range; the precision parameters of pesticide formulations analyzed by LC and S are somewhat poorer. Overall, however, the precision parameters of pesticide formulations are generally independent of analyte, method, and matrix, and are primarily a function of concentration. The method-acceptability decisions of the AOAC for pesticide formulations during the past 75 years can be approximated retrospectively by using a criterion for RSDR that is less than 2 times the RSDR calculated from the Horwitz equation.     

二氧化硫胁迫诱导拟南芥NIT2基因DNA甲基化修饰

DNA甲基化是一种重要的表观遗传修饰形式。利用亚硫酸氢盐修饰后测序法和甲基化敏感性限制性内切酶-PCR（MSRE-PCR）法,研究SO2胁迫对拟南芥腈水解酶（NIT2）基因序列中胞嘧啶甲基化状态的影响,分析甲基化特征改变在植物胁迫应答过程中的作用。研究发现,30 mg.m-3的SO2连续熏气3 d后,拟南芥植株地上组织细胞中NIT2基因启动子区域CG和CHH（H为C,A或T）位点甲基化水平下降,总甲基化水平降低,但未检出编码区5′端目的片段中CCGG位点甲基化状态的改变。RT-PCR分析表明,SO2胁迫组拟南芥植株地上组织细胞中NIT2基因的转录水平高于对照组。研究结果表明,SO2胁迫导致拟南芥NIT2基因启动子区甲基化水平降低,NIT2基因转录上调,说明SO2胁迫能诱发拟南芥基因胞嘧啶甲基化水平改变,启动子区甲基化水平的降低可能与防御基因的诱导表达有关,胞嘧啶甲基化修饰参与了植物的抗逆生理过程。     

Influence of arbuscular mycorrhizal fungi,inoculum level and phosphorus placement on growth and phosphorus uptake of <Emphasis Type="Italic">Phyllanthus calycinus</Emphasis> under jarrah forest soil

Phosphorus uptake and biomass production may vary between arbuscular mycorrhizas (AM) species of contrasting life cycles and their modes of interaction with host plants. This research investigated differences among three AM fungi from different genera isolated from the jarrah forest with respect to their colonisation, biomass production and P uptake over time. The understorey plant Phyllanthus calycinus, has been shown to be extensively mycorrhizal. We examined differences in the capacity of fungi associated with this plant to access a P point source from increasing distances from the root. The methodology simulated “pockets” of P in forest soil. Phosphorus sources in root-exclusion mesh bags were inserted 2, 4 and 6 cm from plant roots restrained in separate mesh bags. Acaulospora laevis colonised plants to the greatest extent, producing higher plant biomass and P uptake compared to the other two fungi. Plant biomass and P uptake were enhanced where P sources were closest to the root, as expected, but there were differences among fungi in response to level of inoculum used for each fungus. The capacity of individual AM fungi to access a point source of P at different distances varied in a highly mycorrhizal native forest understorey plant in parallel with plant growth and P uptake.     

Evaluation of CYP1A Protein Induction, Biotransformation Capacity and DNA Adduct Formation in a Rat Hepatoma Cell Line (Fao), as Biomarkers of Organic Contamination in Environmental Soil Samples

Induction of cytochrome P4501A (CYP1A) immunopositive protein was evaluated in the rat hepatoma cell line Fao, as a biomarker of organic pollution in extract of environmental soil samples, exposed to different sources and degrees of chemical contamination. Soil samples were collected in one area in Russia (Monchegorsk) and two areas in Southern Norway (Fiskaa and Birkenes). In addition, one reference soil sample was collected in Central Norway (Høoylandet). Contents of selected polycyclic aromatic hydrocarbons (PAHs) in the samples were also evaluated. To further evaluate the inducibility of the most potent soil extract (Fiskaa), S9 fraction of Fao cells, pretreated with this extract, was used as an activation system in the Ames Salmonella assay. The DNA adduct forming capacity of the soil extracts, analyzed by the ³²P-postlabeling technique, was also investigated in Fao cells. The Fao cell line has previously been found to be a very sensitive biomonitoring system, that responds to environmentally relevant concentrations of planar model contaminants with increased level of CYP1A immunopositive protein and DNA adducts. In the present study the Fao cell line also showed its potential for use in evaluating the CYP1A inducing potency of environmental samples. All soil extracts induced CYP1A protein in the Fao cells, although the level of induction varied between the soil samples. The Fiskaa soil extract was the most potent CYP1A inducer and this extract also contained the highest level of PAHs. No significant correlation was observed between the level of the total of 16 PAHs and CYP1A protein level. However, a significant correlation was observed between CYP1A protein level and the level of Benzo[a]pyrene (B[a]P), which is a very potent CYP1A inducer. The S9 fraction of pretreated Fao cells activated B[a]P to mutagens in a concentration-dependent relationship, although the response was weak. No DNA adducts were detected in cells exposed to the soil extracts. This demonstrates the necessity of determining several biomarker parameters simultaneously as one single biomarker may fail to respond to potentially harmful compounds.     

VA菌根对蚕豆吸收钼、磷营养的研究

在肥力较低的灰棕紫泥土上,研究了VA菌根(Glomus epigaeum)对蚕豆(Vicia faba)的钼、磷营养及其效应。盆裁试验结果表明,接种VA菌根不仅有利于植物对磷的吸收,而且还有利于钼的吸收。并能促进根系生长和根瘤形成,进而促进地上部分生长,植株健壮,抗赤斑病能力和抗衰老能力增强,从而使产量和品质等都优于未接种VA菌根处理的植株。其中以Mo+P+VA处理的植株最好,Mo+VA处理的植株次之.     

Soil–root cross‐talking: The role of humic substances

The biological activity of humic substances (HS) has been elucidated in the last 40 years. Growth enhancement from HS has been demonstrated in several plants in the laboratory and the field. Morphogenesis effects have also been investigated and include induction of lateral root formation and root hair initiation in intact plants and stimulation of root and shoot development in treated cell calluses. HS enhance nutrient use efficiency, aiding assimilation of both macro and microelements and promoting plant growth by the induction of carbon, nitrogen, and secondary metabolism. The review aims are to: (1) shed light on the mechanism by which plants “talk” with soil through humic substances, (2) elucidate the plant responses to the stimulatory effects of HS, the regulatory circuits that allow plants to cope with humus, and the feedback between plant community structures, and (3) show (in light of recent debate about the alkaline extraction of soil humic substances) the plant capability to acquire biologically active substances from soil. It will be shown that plants modify soils, creating and maintaining favorable habitats for growth and survival. Therefore, organic substances exuded by roots are not a wasteful loss of carbon and energy. They represent an evolved strategy by which plants “talk” to the soil. The mobilization of bioactive organic/humic substances from bulk soil or bulk humus is critical to plant and soil health.     

低氮胁迫下水稻根系的发生及生长素的响应

采用水培实验,研究了5个氮(N)浓度下(0.01～5 mmol L-1)水稻的生物量、体内氮浓度、根系发育、体内生长素浓度以及生长素外流蛋白OsPIN家族基因的表达情况。结果表明,与正常供氮水平(2.5mmol L-1)相比,低氮(0.01 mmol L-1)胁迫下水稻根冠比增加28%,地上部全氮浓度降低约20%,根系全氮浓度降低约33%,种子根长度增加25%,种子根上的侧根密度降低26%,倒一叶中的生长素含量增加140%,而根茎结合处和根系的生长素浓度分别下降22%和60%;RT-PCR的结果表明,低氮(0.01 mmol L-1)胁迫下水稻根系中OsPIN1a-b、OsPIN2、OsPIN5a-b和OsPIN9基因表达显著下调;而外源生长素α-萘乙酸(NAA)和生长素极性运输抑制剂1-萘氨甲酰苯甲酸(NPA)的施加均能影响到水稻种子根长和种子根上的侧根密度。由此推论,低氮胁迫下水稻体内生长素从倒一叶到根系极性运输减少是水稻根系对低氮胁迫响应的生理机制之一。     

Influence of soil inoculation with vesicular-arbuscular mycorrhiza and a phosphate-dissolving bacterium on plant growth and 32P-uptake

The influence of inoculation of soil with a vesicular-arbuscular mycorrhizal fungus (Glomus fasciculatus) and a phosphate-dissolving bacterium (Bacillus circulans) on phosphate solubilization, growth of finger millet (Eleusine coracana) and phosphorus uptake from ³²P-labelled tricalcium phosphate and superphosphate were studied. The mycorrhizal plants produced more dry matter and removed more ³²P from the soil than non-mycorrhizal plants, but did not show increased ³²P activity per unit plant mass. The 30 mm NH₄F-HCl extractable ³²P (available ³²P) in soil, plant ³²P activity and total P uptake were enhanced by soil inoculation with the bacterium. In the treatment receiving both inocula a synergistic effect was recorded with increased P uptake and dry matter production.     

Oxygreen process applied on nongerminated and germinated wheat: role of hydroxamic acids

Wheat samples were taken at different stages of germination characterized by their falling number (which is a relevant indicator of germination) from 400 to 60 s. Each batch was treated by the Oxygreen process, a treatment by ozone, in a closed sequential batch reactor. Leucotriene B4 (LTB 4) was induced by germination, but ozone treatment did not increase this effect. Extract obtained from these wheat batches was applied on human epithelial bronchial cells. Wheat extract from nongerminated wheat did not induce any DNA adduct. More the wheat germination gets underway, more DNA adducts are observed. In contrast, germination did not affect the cell viability. Ozonization of wheat exemplified genotoxic effects only if the wheat was germinated. The implication of hydroxamic acids is discussed. In conclusion, ozonization of wheat, of high milling quality, does not pose any problem.     

Reversible and covalent binding of 5-(hydroxymethyl)-2-furaldehyde (HMF) with lysine and selected amino acids

The chemical reactivity of 5-(hydroxymethyl)-2-furaldehyde (HMF) with lysine, glycine, and proline was studied using isotope labeling technique. To confirm the formation of HMF adducts in glucose amino acid model systems, a useful strategy was developed in which products simultaneously possessing six glucose (HMF moiety) and any number of amino acid carbon atoms in addition to nitrogen were targeted using specifically labeled precursors such as [(15)N(α)]lysine·2HCl, [(15)N(ε)]lysine·2HCl, [U-(13)C(6)]lysine·2HCl, [(13)C(6)]lysine·2HCl, and [U-(13)C(6)]glucose in the case of lysine model system. In addition, model systems containing HMF and amino acids were also studied to confirm specific adduct formation. Complete labeling studies along with structural analysis using appropriate synthetic precursors such as HMF Schiff base adducts of piperidine and glycine have indicated that HMF generated in the glucose/amino acid model systems initially forms a Schiff base adduct that can undergo decarboxylation through an oxazolidin-5-one intermediate and form two isomeric decarboxylated Schiff bases. Unlike the Schiff bases resulting from primary amines or amino acids such as glycine or lysine, those resulting from secondary amino acids such as proline or secondary amines such as piperidine can further undergo vinylogous Amadori rearrangement, forming N-substituted 5-(aminomethyl)furan-2-carbaldehyde derivatives.