Supramolecular interactions of humic acids with organic and inorganic xenobiotics studied by capillary electrophoresis

Methodology based on capillary electrophoresis (CE) to study humic acids (HAs)-xenobiotics interactions is proposed. The interactions of HAs with organic and inorganic xenobiotics like paraquat, diquat, p,p(')-DDE, p,p(')-DDT, potassium ferrocyanide, potassium ferricyanide, chloride, 4-nitrocatechol and other organic compounds were studied. They were found to be of different kind depending on the structure of the xenobiotic molecule and on its charge (neutral, positive or negative). Ion binding, hydrogen bonding, van der Waals forces, ligand exchange, hydrophobic and hydrophilic adsorption, charge-transfer complexes and sequestration are some of the different mechanisms proposed to bind inorganic and organic compounds to HAs.It was also observed that some of the pollutants are strongly complexed (bound) only with some of the HA fractions forming quite stable entities of supramolecular kind, which can migrate independently. In addition, the stability constant of HA-Cl (negatively charged species) was estimated to be logk=3.1+/-0.95. In order to explain the interaction between negatively charged HAs and inorganic anions (like Cl(-), [Fe(CN)(6)](3-) and [Fe(CN)(6)](4-)), it is proposed that macropolycyclic polyamine structures are present in HA supramolecules, and that they are responsible for such strong binding.     

Hydroxycinnamic acid derivatives in an aquatic liverwort as possible bioindicators of enhanced UV radiation

We examined, under laboratory conditions, the physiological responses of the aquatic liverwort Jungermannia exsertifolia subsp. cordifolia to artificially enhanced ultraviolet (UV) radiation for 82 days, especially considering the responses of five hydroxycinnamic acid derivatives. This species lives in mountain streams, where it is exposed to low temperatures and high UV levels, and this combination is believed to increase the adverse effects of UV. Enhanced UV radiation hardly caused any change in several physiological variables indicative of vitality, such as Fv/Fm and chlorophylls/phaeopigments ratio (OD430/OD410). Thus, this liverwort seemed to be tolerant to UV radiation, probably due to the accumulation of three UV-absorbing hydroxycinnamic acid derivatives: p-coumaroylmalic acid, 5'-(7',8'-dihydroxycoumaroyl)-2-caffeoylmalic acid, and 5'-(7',8'-dihydroxy-7-O-beta-glucosyl-coumaroyl)-2-caffeoylmalic acid. These compounds might serve as bioindicators of enhanced UV radiation.     

Polybrominated diphenyl ethers (PBDEs) and their methoxylated derivatives in pike from Swedish waters with emphasis on temporal trends, 1967-2000

Temporal trends of five tetra- to hexabromodiphenyl ethers [BDE47, BDE99, BDE100, BDE153 and BDE154) and two methoxy-tetraBDEs [6-methoxy-2,2',4,4'- tetraBDE (6-MeO-BDE47) and 2'-methoxy-2,3',4,5'- tetraBDE (2'-MeO-BDE68)] in pike from Lake Bolmen for the years 1967-2000, are presented. All BDE congeners show increasing trends up to the mid-1980s (Sigma5PBDE from 60 to 1600 pg/g wet weight in 1989, i.e. a more than 25-fold increase), and then decrease or level off. The decreasing trends of PBDEs after the 1980s were considerably slower in the present study than was found in a study of an environmental matrix from the Baltic Proper covering the same time period. This difference suggests local sources near Lake Bolmen. The MeO-BDEs show initially decreasing concentrations, which for 6-MeO-BDE47 continues until the early 1990s. The concentrations of 6-MeO-BDE47 in herring from five locations along the Swedish coast increased from south to north in the Baltic Sea. No correlation between the concentrations of the BDE congeners and the MeO-BDEs was observed, indicating sources other than PBDEs for these compounds. The presence of MeO-BDEs in fish from lakes with different characteristics suggests a natural production not favoured by eutrophication, or dependent on sampling season and geographical location.     

The effects of pH, bromide and nitrite on halonitromethane and trihalomethane formation from amino acids and amino sugars

In this study, the effects of pH, bromide and nitrite on the formation of halonitromethanes (HNMs) and trihalomethanes (THMs) from eight amino acids (glycine, alanine, serine, cysteine, aspartic acid, glutamic acid, lysine and histidine) and four amino sugars (glucosamine, galactosamine, N-acetylglucosamine and N-acetylneuraminic acid) were examined for chlorination and ozonation followed by chlorination. During ozonation-chlorination, two amino acids, glycine and lysine, exhibited distinctly higher HNM formation than the other compounds. The formation of HNMs was higher at pH 8 than 6. Glycine and lysine also produced higher levels of THMs than the other compounds at pH 8. The presence of nitrite resulted in an increase in HNM formation. The presence of bromide increased the HNM formation, especially brominated HNM species. Bromine incorporation factors of trihalogenated HNMs were higher than those of THMs. For chlorination alone, HNM levels were about the detection limit (4 nM or 0.7 μg L⁻¹) at pH 6 and 8, and in the presence of bromide or nitrite. Amino acids and amino sugars tested, except glycine and lysine, showed relatively low levels of THM (∼15 μg L⁻¹) formation.     

Levels and tissue-dependent distribution of dioxin in Japanese domestic leafy vegetables--from the 1999 national investigation

In 1999, Japanese domestic leafy vegetables were successively investigated for levels of dioxins, including 17 dibenzo-p-dioxins/dibenzofurans (PCDD/Fs), four non-ortho co-planar PCBs (co-PCBs) and eight mono-ortho co-PCBs, all of which had been assigned toxic equivalency factors (TEFs) by WHO in 1997. The mean levels of dioxins in the edible portions were 0.07 (0.09) pg TEQ/g in spinach, 0.13 (0.14) pgTEQ/g in garland chrysanthemum, 0.01 (0.04) pg TEQ/g in mitsuba (marsh parsley) and 0.01 (0.03) pg TEQ/g in chingentsuai (Brassica Campestris var. chinesis), when non-detects were set to zero (and set to half the limit of detection). In order to understand the dioxin pollution of leafy vegetables in detail, a further investigation of dioxin levels in the tissues of spinach was conducted. As a result, the dioxin levels in the leaves were found to be higher than those in the stem and red collar, but they were much lower than those found in the primary and secondary roots, which are considerably affected by the soil, which is recognized as a sink of airborne dioxins. The dioxin levels in edible portions (leaves, stem and red collar) were obviously lower than those in non-edible portions (primary and secondary roots). In addition, from the finding that several lower-chlorinated PCDD/Fs and co-PCBs, namely 2,3,7,8-TCDD/F, 1,2,3,7,8-PeCDD, 1,2,3,7,8/2,3,4,7,8-PeCDFs, 1,2,3,7,8,9-HxCDD, 1,2,3,4,7,8-/1,2,3,6,7,8-HxCDFs, 3,3',4,4'-TCB, 2,3,3',4,4'-/2,3',4,4',5-PeCBs, and 2,3',4,4',5,5'-HxCB, were more highly represented in the dioxins in the leaves than in those in the secondary roots, it was suggested that in leafy vegetables the deposition of gaseous, presumably moderately volatile dioxins in leaf wax is another pollution pathway in addition to the adhesion of dioxin-contaminated particles including soil.     

Quantification of nitrogen in the liquid fraction and in vitro assessment of lysine bioavailability in the solid fraction of soybean meal hydrolysates

Soybean meal (SBM) is a product generated from the manufacture of soybean oil and has the potential for use as a source of fermentable sugars for ethanol production or as a protein source for animal feeds. Knowing the levels of nitrogen available from ammonium is a necessary element of the ethanolic fermentation process while identifying the levels of essential amino acids such as lysine is important in determining usage as a feed source. As such the purpose of this study was to quantify total nitrogen and ammonium in the liquid fraction of hydrolyzed SBM and to evaluate total and bioavailable lysine in the solid fraction of the hydrolyzed SBM. The effects of acid concentration, cellulase and β-glucosidase on total and ammonium nitrogen were studied with analysis indicating that higher acid concentrations increased nitrogen compounds with ammonium concentrations ranging from 0.20 to 1.24 g L^?1 while enzymatic treatments did not significantly increase nitrogen levels. Total and bioavailable lysine was quantified by use of an auxotrophic gfpmut3 E.coli whole-cell bioassay organism incapable of lysine biosynthesis. Acid and enzymatic treatments were applied with lysine bioavailability increasing from a base of 82% for untreated SBM to up to 97%. Our results demonstrated that SBM has the potential to serve in ethanolic fermentation and as an optimal source essential amino acid lysine.     

Biodegradation of polyaromatic hydrocarbons by recombinant bacteria containing Alasan gene

Alasan, the bioemulsifier of Acinetobacter radioresistens KA53, is a high molecular-mass complex of an alanine-containing polysaccharide and three proteins. This gene was isolated from the A. radioresistens using PCR and the amplified product (850 bp) was cloned using PGEMT cloning kit. DNA sequence was carried out for the amplification and sequence analysis showed that the PCR product was highly similar to the OMPA precursor protein which called Alan. Subcloning was carried out into pTRAC expression vector and the purified protein was used in mineralisation of the polyaromatic hydrocarbon. As well as the recombinant E. coli cells were cultured directly on MSM containing four polyaromatic hydrocarbon using real time method (phenol, naphthalene, phenantherene and antherathene). The obtained results showed that no difference between the purified protein and the using of the recombinant cells directly with the presence of the inducer like IPTG. Both of the purified protein and the recombinant cells showed high degradation rates for (anthrathene and phenantherene).     

Identification of hydroxylated metabolites in 2,2',4,4'-tetrabromodiphenyl ether exposed rats

Faeces from day 1-5 of orally administered 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) in rat have been analysed for hydroxylated metabolites. Six hydroxylated tetrabrominated diphenyl ethers, as well as three hydroxylated tribrominated diphenyl ethers found, were structurally identified. They were 2'-hydroxy-2,4,4'-tribromodiphenyl ether, 3'-hydroxy-2,4,4'-tribromodiphenyl ether, 4'-hydroxy-2,2',4-tribromodiphenyl ether, 6-hydroxy-2,2',4,4'-tetrabromodiphenyl ether, 2'-hydroxy-2,3',4,4'-tetrabromodiphenyl ether, 3-hydroxy-2,2',4,4'-tetrabromodiphenyl ether, 5-hydroxy-2,2',4,4'-tetrabromodiphenyl ether, 4'-hydroxy-2,2',4,5'-tetrabromodiphenyl ether and 4-hydroxy-2,2',3,4'-tetrabromodiphenyl ether. The analysis was performed using gas chromatography-mass spectrometry (GC-MS). The identification of the hydroxylated polybrominated diphenyl ether (OH-PBDE) metabolites in the rat faeces was supported by similar relative retention times (RRTs) versus 2,2',3,4,4',5-hexabromodiphenyl ether (BDE-138) on two columns of different polarities compared to the authentic references. The identification of the OH-PBDE metabolites was also supported by full scan electron ionisation mass spectra. Two of the identified OH-PBDE metabolites have identical structures as natural products, which previously have been isolated from marine sponges and an ascidian.     

超强静磁场对染料脱色菌Escheriehia coli的acpD基因的影响

将大肠杆菌E.coli ATCC8739置于12.0 T超强静磁场(ultra-strong static magnetic field,SMF)中进行处理,获得了磁场处理0.5、1、2、4和8 h的菌株E.coli-SMFn(n=0.5、1、2、4、8)。在37℃、pH 7、静置的条件下,菌株对偶氮染料AR14(I.C.Acid Red 14)的脱色结果指出,当反应进行到4、6和8 h时,E.coli-SMF8的脱色效率分别为18%、55%和96%,远高于E.coli ATCC8739的3%、19%和50%,表明SMF作用显著地增强了菌株的脱色效率。基因组DNA提取、PCR扩增、分子克隆以及基因测序的实验结果进一步表明,全部6例E.coli ATCC8739菌株的偶氮还原酶基因(acpD)序列均与GenBank中报道的完全一致;而E.coli-SMF8菌株的acpD-SMF8核酸序列中缺失了第99位的胞嘧啶。该缺失突变不仅使acpD-SMF8的核酸序列与acpD的存在显著不同,同时得到了具新活性中心的偶氮还原酶AzoR-SMF8。这个新的活性中心具有更强的黄素(FMN)结合能力,因此使该酶与偶氮染料的亲和力大大增加,促进了脱色效率的提高。     

Formation of mutagens by photolysis of amino acids in neutral aqueous solution containing nitrite or nitrate ion

Nine amino acids, i.e., alanine, threonine, cysteine, glutamic acid, arginine, proline, tryptophan, phenylalanine and tyrosine, were irradiated with UV light in water containing nitrite or nitrate ion under neutral conditions. The mutagenicities of the ether extracts and the residual water layers of the reaction mixtures were assayed with and without S-9 mix using $\text{Salmonella typhimurium}$ TA98 and TA100. Three aromatic amino acids, tryptophan, phenylalanine and tyrosine, were found to give direct-acting and frameshift mutagens by irradiation in aqueous nitrite solution. Among them, the ether extract of tryptophan exhibited the strongest mutagenicity toward TA98. In the case of irradiation in aqueous nitrate solution, only the ether extract of tryptophan exhibited weak mutagenicity toward TA98 without S-9 mix. The effects of nitrite concentration, irradiation time and pH on mutagen formation from tryptophan and some characteristics of the produced mutagens were examined.     

无机抗菌剂在建筑涂料中的应用研究

研究了5种无机抗菌剂:锌型沸石抗菌剂、银型沸石抗菌剂、氧化锌晶须复合抗菌剂、纳米TiO2光催化抗菌剂和Ca10-x-yTixZny(PO4)6(OH)2复合抗菌剂在涂料中的应用。以对大肠杆菌(E.coli)的灭菌效果评价抗菌性能。结果表明:锌型抗菌涂料的抗菌效果较差,后4种类型抗菌涂料具有很好抗菌效果。但是,银型抗菌剂和氧化锌晶须复合抗菌剂在涂料中存在易引起涂料变色和难分散问题;纳米TiO2光催化型抗菌剂在涂料中易加快涂料老化;而Ca10-x-yTixZny(PO4)6(OH)2复合抗菌剂既具有良好的抗菌性能,又对涂料的基本性能没有影响,它能够应用到抗菌涂料中。     

Adsorption and desorption variability of four herbicides used in paddy rice production

This investigation was performed to determine the effect of physicochemical soil properties on penoxsulam, molinate, bentazon, and MCPA adsorption-desorption processes. Four soils from Melozal (35° 43' S; 71° 41' W), Parral (36° 08' S; 71° 52' W), San Carlos (36° 24' S; 71° 57' W), and Panimavida (35° 44' S; 71° 24' W) were utilized. Herbicide adsorption reached equilibrium after 4 h in all soils. The Freundlich L-type isotherm described the adsorption process, which showed a high affinity between herbicides and sorption sites mainly because of hydrophobic and H-bonds interaction. Penoxsulam showed the highest adsorption coefficients (4.23 ± 0.72 to 10.69 ± 1.58 mL g?1) and were related to soil pH. Molinate showed K(d) values between 1.72 ± 0.01 and 2.3 ± 0.01 mL g?1 and were related to soil pH and organic matter, specifically to the amount of humic substances. Bentazon had a high relationship with pH and humic substances and its K(d) values were the lowest, ranging from 0.11 ± 0.01 to 0.42 ± 0.01 mL g?1. MCPA K(d) ranged from 0.14 ± 0.02 to 2.72 ± 0.01 mL g?1, however its adsorption was related to humic acids and clay content. According to these results, the soil factors that could explain the sorption process of the studied herbicides under paddy rice soil conditions, were principally humic substances and soil pH. Considering the sorption variability observed in this study and the potential risk for groundwater contamination, it is necessary to develop weed rice management strategies that limit use of herbicides that exhibit low soil adsorption in areas with predisposing conditions to soil leaching.     

Estrogenic and thyroid hormone activity of a series of hydroxy-polychlorinated biphenyls

A series of novel synthetic monohydroxy polychlorinated biphenyls (OH-PCBs) (5 trichloro-, 5 tetrachloro- and 5 pentachloro-compounds) have been characterized (1H and 13C NMR and high resolution MS) and their estrogenic and thyroid hormone activities assessed using a yeast two-hybrid assay, both with and without possible metabolic activation by rat liver S9 preparation. Moderate estrogenic activity was found for 2,3,4(')-trichlorobiphenyl-4-ol (compound 5) but this was eliminated when exposed to the S9 mix. 2,2('),3('),4,6-Pentachlorobiphenyl-3-ol (13) and 2('),3,3('),6-tetrachlorobiphenyl-4-ol (10) both showed weak estrogenicity in the absence of the S9 mix. The estrogenicity of compound (10) was enhanced 10-fold by exposure to S9 metabolic activation but that of compound (13) remained unchanged. 2('),4,5('),6-Tetrachlorobiphenyl-2-ol (6) showed strong thyroid hormonal activity (5% of that of T4) whereas 3('),4,6-trichlorobiphenyl-3-ol (4), compound (10) and 2,3('),4,5('),6-pentachlorobiphenyl-3-ol (14) showed moderate activity, and 2('),3,3('),5-tetrachlorobiphenyl-2-ol (8) and 3,3('),5,5('),6-pentachlorobiphenyl-2-ol (11) showed weak activity. The activity of (4) was eliminated by S9 metabolic activation whereas those of (6) and (14) were weakened and that of (10) remained unchanged.     

Potential environmental influence of amino acids on the behavior of ZnO nanoparticles

The fate of nanomaterials when they enter the environment is an issue of increasing concern and thus it is important to know how they interact with natural organic molecules since this may have a significant impact on the particles' behavior. Because of our poor knowledge in this regard, the interaction of ZnO nanoparticles with amino acids of contrasting surface charge, including Histidine (HIS), Glycine (GLY), Aspartic acid (ASP) and Glutamic acid (GLU) which occur commonly in natural habitats, such as the plant root zone, was investigated over a range of pH conditions and concentrations. The addition of the individual amino acid led to significant changes in nanoparticle colloidal zeta potential stability, particle size distribution and the extent of agglomeration. Variations in pH resulted in considerable changes in nanoparticle surface charge and hydrodynamic size. In general, the particle size distribution decreased as the amino acid concentration increased, with more acidic conditions exacerbating this effect. In addition, increased concentrations of amino acids resulted in more stable nanoparticles in aqueous suspensions. Histidine had the greatest effect on colloidal stability, followed by Glycine, Aspartic acid and finally Glutamic acid. This study illustrates how nanoparticle behavior may change in the presence of naturally occurring amino acids, an important consideration when assessing the fate of nanoparticles in the environment. Additionally, utilization of amino acids in industrial processes could reduce particle agglomeration and it could lead to a way of employing more sustainable reagents.     

Application of a pathogenicity marker found in Escherichia coli for the assessment of irrigation water quality.

A polymerase chain reaction (PCR)-based method was developed to differentiate between pathogenic and nonpathogenic Escherichia coli (E. coli). A pathogenicity marker, linked to the deletion of the ygfB gene, was identified in 80% of the clinical E. coli isolates tested. This marker, combined with the malic acid dehydrogenase gene, formed the duplex PCR that was subsequently used to screen E. coli isolates recovered from two secondary wastewater treatment plants (STPs) and a river site. All waters samples are used to irrigate dairy farm pasture in the West Gippsland region of Victoria, Australia. Results from three consecutive months of sampling (December 2001 and January and February 2002) indicated that Longwarry STP showed 8, 8, and 0% pathogenic E. coli; Pakenham STP showed 0, 12.5, and 33%; and the Bunyip river site showed 20, 12, and 25% respectively.     

Synthesis of polybrominated diphenyl ethers via symmetrical tetra- and hexabrominated diphenyliodonium salts

Polybrominated diphenyl ethers (PBDEs) are a class of brominated flame retardants (BFRs) which have become widespread environmental pollutants due to their persistence and bioaccumulativeness. Pure authentic PBDE congeners are required for chemical analysis, assessments of their chemical/physical properties and toxicological studies. We here report an improved method for synthesis of authentic PBDE congeners applying bromophenols and symmetrical brominated diphenyliodonium salts as building blocks. Altogether, 13 PBDEs were synthesized of which seven are new. The improved coupling reaction between the bromophenol and the brominated diphenyliodonium salts resulted in enhanced yields for PBDEs substituted with more than six bromine atoms. Also, improvements in iodonium salt synthesis made it possible to synthesize symmetrical hexabromodiphenyliodonium salts for the first time, i.e. 2,2',3,3',4,4'-, 2,2',4,4',5,5'- and 2,2',4,4',6,6'-hexabromodiphenyliodonium salts and they made it possible to prepare octabrominated PBDEs via the actual coupling method. All synthesized compounds were characterized by (1)H NMR, (13)C NMR spectra and by their melting points. Also, all products except for the diphenyliodonium salts were characterized by mass spectra in electron ionization mode.     

一种未见报道的厌氧氨氧化菌分子生物学鉴定

以(NH4):SO4和NaNO2作为基质,富集厌氧氨氧化污泥.提取厌氧氨氧化污泥中细菌总DNA,纯化后使用特异性引物对厌氧氨氧化菌16S rDNA进行PCR扩增.扩增产物连接到pMD19-T载体,将载体转化到感受态细胞大肠杆菌JM109中,并对其16S rDNA基因进行测序.将测序结果进行系统发育树分析,发现富集得到的厌氧氨氧化菌与Candidatus Anammoxoglobus propionicus进化关系比较接近,是一种尚未见报道的厌氧氨氧化菌.     

Deviations in hepatic amino acid profiles of mouse following repeated hexachlorophene administration

Effects of repeated administration of hexachlorophene (HCP) on the food consumption, body and liver weights, liver somatic indices and hepatic levels of the free amino acids and amino acid-enzyme activities have been studied. A dose-dependent reduction in body weights and food consumption and increased liver somatic indices suggested a progressive and on toward response to HCP. The concentrations of hepatic free amino acids: phenylalanine, tyrosine, leucine, isoleucine, valine, threonine, asparagine, glutamic acid and glutamine were significantly (p less than 0.001) increased while serine, glycine, taurine and alanine levels were unaffected. The aspartic acid levels showed a conspicuous decrement. The specific activity patterns of the amino acid-enzymes denoted reduced oxidation of amino acids in liver. The abnormal rise in branched chain and aromatic amino acids could be due to the reduced utilization because of the depletion of skeletal muscle mass and hepatic dysfunction in HCP intoxication. A pronounced reduction in aspartic acid levels may deserve consideration in the manifestation of hyperammonemia and neurologic symptoms in HCP intoxication.     

Responses of free amino acids in rice seedlings during cyanide metabolism

Responses of free amino acids to botanical assimilation of free cyanide were investigated. Young rice seedlings (Oryza sativa L. cv. XZX 45) were grown in nutrient solution amended with free cyanide (KCN). Cyanide was analyzed in solution as well as in plant materials to estimate the phyto-assimilation potential. Free amino acids in different parts of plants were also measured to determine metabolic responses to KCN exposure. Phyto-assimilation of KCN was obvious, and the rates were positively correlated to the concentration supplied. Although changes in total amino acid content in plant materials were negligible during KCN metabolism (p?>?0.05), responses of different amino acids to KCN treatments were quite different. All treatments with KCN increased the content of proline (Pro) and isoleucine (Ile) in roots significantly compared with control (p?<?0.05), while changes of aspartic acid, lysine, and histidine in roots were more evident at higher KCN treatments (p?<?0.05). Results indicate that the content of Pro, Ile, and tyrosine showed pronounced increase in shoots of rice seedlings exposed to KCN at 1.44 mg CN/L or higher (p?<?0.05). Other amino acids slightly changed in all plant materials exposed to KCN (p?>?0.05). Results indicate that specific amino acids in rice seedlings showed positive response to non-toxic concentrations of exogenous KCN. These findings could provide additional insights into the inducible mechanisms underlying the involvement of amino acids in KCN metabolism.     

Degradation of indulin, a kraft pine lignin, by Serratia marcescens

Serratia marcescens isolated from decaying coconut pith exhibited high lignolytic activity. Growth on indicator medium, analysis of residual indulin, and infra-red spectroscopic analysis indicated the lignolytic potential of the isolate. Ortho-Coumaric acid, ferulic acid, 2,3-dihydroxy cinnamic acid and protocatechuic acid were identified as intermediates involved in indulin degradation by S. marcescens. Qualitative confirmation and quantitative estimation of the intermediates were carried out by high performance thin layer chromatography (HPTLC).