Estrogenicity and acute toxicity of selected anilines using a recombinant yeast assay

Suspected estrogen modulators include industrial organic chemicals (i.e., xenoestrogens), and have been shown to consist of alkylphenols, bisphenols, biphenylols, and some hydroxy-substituted polycyclic aromatic hydrocarbons. The most prominent structural feature identified to be important for estrogenic activity is a polar group capable of donating hydrogen bonds (i.e., hydroxyl) on an aromatic system. The present study was undertaken to explore the estrogenic activity and acute toxicity of chemicals containing a weaker hydrogen bond donor group on aromatic systems, i.e., the amino substituent. There is a great deal of chemical similarity between aromatic amines (anilines) and aromatic alcohols (phenols). The chemicals chosen for the current study contained an amino-substituted benzene ring with hydrophobic constituents varying in size and shape. Thus, 37 substituted aromatic amines were assayed for estrogenic activity EC50 and acute toxicity LC50 using the Saccharomyces cerevisiae recombinant yeast assay. While the EC50 of 17-beta-estradiol occurs at the 10(-10) range, the aniline with the greatest activity had an EC50 of 10(-6) M. Thus, anilines, in general, are capable only of very weak estrogenic activity in this assay. A comparison of estrogenic potency between the present group of anilines and a set of previously tested analogous phenols indicated that anilines are consistently less estrogenic than phenols. A comparison of hazard indices (EC50/LC50) of these chemicals revealed that, for the vast majority of anilines, the EC50 and LC50 were in the same order of magnitude. More specifically, estrogenic activity of para-substituted alkylanilines increases with alkyl group size up to 5 carbons in length, after which the acute toxicity of the larger alkyl-substituents precluded the ability of the compound to induce the estrogenic response.     

Structure-activity relationships and response-surface analysis of nitroaromatics toxicity to the yeast (Saccharomyces cerevisiae)

Inhibition of growth of the yeast Saccharomyces cerevisiae (Cmiz, the minimum concentration that produced a clear inhibition zone within 12 h) for 24 nitroaromatic compounds was investigated and a quantitative structure-activity relationship (QSAR) developed based on hydrophobicity expressed as the l-octanol/water partition coefficient in logarithm form, log K(ow), electrophilicity based on the energy of the lowest unoccupied orbital (E(lumo)). All nitrobenzene derivatives exhibited enhanced reactive toxicity than baseline. The toxicities of mono-nitrobenzenes and di-nitrobenzenes were elicited by different mechanisms of toxic action. For mono-nitro-derivatives, both significant log K(ow) based and strong E(lumo)-dependent relationships were observed indicating that their toxicities were affected both by the penetration process and the interaction with target sites of interaction. The toxicities of di-nitrobenzenes were greater than mono-nitrobenzenes and no log K(ow)-dependent but highly significant E(lumo)-based relationship was obtained. This suggests that toxicity of di-nitrobenzenes was highly electrophilic and involved mainly their in vivo electrophilic interaction with biomacromolecules. In an effort to model the elevated toxicity of all nitrobenzenes, a response-surface analysis was performed and this resulted in a highly predictive two-variable QSAR without reference to their exact mechanisms (Cmiz = 0.41 log K(ow) - 0.89 E(lumo) - 0.46, r2 = 0.87, Q2 = 0.86, n = 24).     

Yeasts as a model for assessing the toxicity of the fungicides Penconazol, Cymoxanil and Dichlofluanid

In the present work the sensitivity of yeast strains of Kluyveromyces marxianus, Pichia anomala, Candida utilis, Schizosaccharomyces pombe and Saccharomyces cerevisiae, to the fungicides cymoxanil, penconazol, and dichlofluanid, was evaluated. Dichlofluanid induced the most negative effects, whereas penconazol in general was not very toxic. Overall, our results show that the parameters IC50 for specific respiration rates of C. utilis and S. cerevisiae and C(D) for cell viability of S. cerevisiae can be applied to quantify the toxicity level of the above compounds in yeast. Hence, could be explored as an alternative or at least as a complementary test in toxicity studies and, therefore, its potential for inclusion in a tier testing toxicity test battery merits further research.     

非固定化和固定化啤酒酵母对Cd^2＋和Cu^2＋的吸附特性研究

对比了不同吸附剂对重金属的吸附效果，同时研究了啤酒酵母的固定化方法、菌体用量对吸附效果的影响、非同定化和固定化啤酒酵母吸附热力学特性。研究结果表明，非固定化死啤酒酵母对Cd^2＋的单位菌体吸附量是常用吸附剂活性炭的3倍；由1：3的海藻酸钠与碱处理啤酒酵母（w／w）制得的固定化颗粒吸附效果最好；菌体用量的增加会降低单位菌体对重金属的吸附量；啤酒酵母对重金属的吸附位点有限，Cd^2＋的实际最大吸附量为13．95mg／g，Cu^2＋为7．67μg／g。非固定化和固定化啤酒酵母对Cu^2＋和Cd^2＋的等温吸附过程均可用Linear方程、Langmuir方程和Freundlich方程来进行拟合，但非同定化啤酒酵母以Langmuir方程最优，其拟合计算的最大吸附量qmzxCd和qmxxCu分别为13．96mg／g和8．01mg／g；固定化啤酒酵母以Freundlich方程最优，实际最大吸附量Cd为75．41mg／g，Cu为66．58mg／g。     

含氯代烃废水的生物毒性与处理方法探讨

卤代烃是在有机物分子中的碳原子上，用卤素基团取代出氢的卤化产物，这个变化使有机物的生物毒性增大，这是卤素有机态毒性的体现；另一方面，卤代烃在生物水解或降解过程中，又会重新释放出带正电荷的卤素，与水结合后成为次卤酸而具有无机态卤素的生物毒性。作者在提出这种卤代烃生物毒性学说的基础上，提出了一系列在含卤代烃废水预处理与生物处理中的解毒、降毒、抗毒和减荷及提高可生化性的措施，以提高含卤代烃废水的综合处理效率。     

Biosorption of water-soluble dyes on magnetically modified Saccharomyces cerevisiae subsp. uvarum cells

Brewer's yeast (bottom yeast, Saccharomyces cerevisiae subsp. uvarum) cells were magnetically modified using water based magnetic fluid stabilized with perchloric acid. Magnetically modified yeast cells efficiently adsorbed various water soluble dyes. The dyes adsorption can be described by the Langmuir adsorption model. The maximum adsorption capacity of the magnetic cells differed substantially for individual dyes; the highest value was found for aniline blue (approx. 220 mg per g of dried magnetic adsorbent).     

Polyfluorinated chemicals in a spatially and temporally integrated food web in the Western Arctic

This study reports on an investigation of the presence of polyfluorinated chemicals in a spatially and temporally integrated set of biological samples representing an Arctic food web. Zooplankton, Arctic cod, and seal tissues from the western Canadian Arctic were analyzed for perfluoroalkyl sulfonates [PFAS], perfluorocarboxylates [PFCAs], and other polyfluorinated acids. Perfluorooctane sulfonate [PFOS] was found in all samples [0.20-34 ng/g] and in the highest concentrations. PFCAs from nine to 12 carbons were quantified in most of the samples [0.28-6.9 ng/g]. PFCAs with carbon chain lengths of eight or less were not detected. Likewise, 8-2 fluorotelomer acid [8-2 FTA] and 8-2 fluorotelomer unsaturated acid [8-2 FTUA], products of fluorotelomer environmental transformation, were not detected. 2H,2H,3H,3H, heptadecafluoro decanoic acid [7-3 Acid], an additional metabolite from fluorotelomer biological transformation, was detected only in seal liver tissue [0.5-2.5 ng/g]. The ratios of branched to linear PFOS isomers in fish and seal tissue were not similar and did not match that of technical PFOS as manufactured. No branched PFCA isomers were detected in any samples. It is concluded that differing pharmacokinetics complicate the use of branched to linear ratios of PFCAs in attributing their presence to a specific manufacturing process. A statistical analysis of the data revealed significant correlations between PFOS and the PFCAs detected as well as among the PFCAs themselves. The 7-3 Acid was not correlated with either PFCAs or PFAS, which suggests that it may have a different exposure pathway.     

Design and development of novel insect growth regulators: synthesis, characterization and effect of benzoyl thymyl thioureas and ureas on total haemocyte count of Dysdercus koenigii

Insect-growth regulators (IGRs) have been receiving foremost attention as potential means of selective insect control. Benzoyl phenyl urea (BPU) is a well-known IGR having chitin synthesis inhibitor activity. Mimics of BPU have been synthesized by suitable derivatization of a naturally occurring monoterpenoid, thymol (2-isopropyl-5-methyl phenol) to form a = series of substituted benzoyl thymyl thioureas (BTTUs) [IVa-f] and benzoyl thymyl ureas (BTUs) [Va-f]. The synthesized compounds have been characterized by (1)H and (13)C NMR, LC-MS and elemental analysis. These derivatives have been screened for their effect on total haemocyte count of Dysdercus koenigii. It has been observed that the introduction of substituted benzoyl thiourea and urea linkage into a thymol ring via an amino group results in higher activity than the parent compound thymol and a comparable pattern of results with the standard insect-growth regulators, Penfluron. Urea [Va-f] compounds exhibited greater effect on Total Haemocyte Count (THC) than thiourea [IVa-f]. Fluoro substitution enhanced the effect on THC more than chloro substituted compounds, while ortho-substitution resulted in a better effect than para-substitution. The results described in this paper are promising and provide new array of synthetic chemicals that may be utilized as insect growth regulators.     

A comparative study on the biosorption characteristics of some yeasts for Remazol Blue reactive dye

Biosorption capacities and rates of different kinds of dried yeasts (Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces marxianus, Candida sp., C. tropicalis, C. lipolytica, C. utilis, C. quilliermendii and C. membranaefaciens) for Remazol Blue reactive dye from aqueous solutions were compared under laboratory conditions as a function of initial pH and initial dye concentration. Optimum initial biosorption pH was determined as 2 for all the yeasts. All the yeast species showed comparable and very high dye sorption at 100 mg/l initial dye concentration. The equilibrium sorption capacity of the biomass increased with increasing initial dye concentration up to 400 mg/l for Candida sp. C. lipolytica and C. tropicalis; up to 300 mg/l for C. quilliermendii and C. utilis and up to 200 mg/l for S. cerevisiae, S. pombe, K. marxianus and C. membranaefaciens while the adsorption yield of dye showed the opposite trend for all the yeasts. Among the nine yeast species, C. lipolytica exhibited the highest dye uptake capacity (Q(0) = 250 mg/g). Both the Freundlich and Langmuir adsorption models were found suitable for describing the biosorption of the dye by all the Candida yeasts (except C. membranaefaciens). The results indicated that the dye uptake process followed the pseudo-second-order kinetics for each dye-yeast system.     

Atmospheric BTX and polyaromatic hydrocarbons in Rio de Janeiro, Brazil

Polycyclic aromatic and monoaromatic (benzene, toluene and xylene, or BTX) hydrocarbons were monitored in Rio de Janeiro, Brazil, during the summer of 1998/1999. The levels of these aromatic chemicals decreased with distance from main roads, indicating mobile sources are the main pollutant emitters in this Latin American city. Benzo[ghi]perylene/indeno[1,2,3-cd]pyrene and benzene/toluene ratios corroborate this idea. However, higher benzene/toluene ratios at one of the major access routes into the city suggest pollutant inputs from a nearby refinery. Literature data were reviewed in order to outline differences and similarities among sources and levels of aromatic pollutants in large urban agglomerations worldwide. Concentrations of benzo[a]pyrene and benzene, which are well-known carcinogenic chemicals, were relatively low in Rio de Janeiro. This fact was attributed to specific atmospheric conditions during the tropical summer and differences in vehicle fuel composition.     

Beta-cyclodextrin(beta-CD) influence on the biotoxicities of substituted benzene compounds and pesticide intermediates

The acute toxicities of 19 substituted benzene compounds and intermediates of pesticides to Photobacterium phosphoreum in beta-CD aqueous solution (5 g/L) were studied and compared with those in water through luminescent bacteria toxicity test, and 15min-EC50 values were obtained. The presence of beta-CD significantly reduce biotoxicities of the low-polarity compounds, while for the higher-polarity compounds, such effect was slight. The differences of biotoxicities were correlated with Log10Kow (octanol/water partition coefficients). A significant linear relationship was obtained with some abnormal data picked out. Some explanations concerning molecular structure of chemicals as well as beta-CD were given for the relationship.     

Investigation of photodegradation and hydrolysis of selected substituted urea and organophosphate pesticides in water

Introduction  

Photodegradation and hydrolysis of two substituted urea herbicides, monolinuron [3-(4-chlorophenyl)-1-methoxy-1-methylurea] and linuron [3-(3,4-dichlorophenyl)-1-methoxy-1-methylurea], and one organophosphorous insecticide, phoxim [2-(diethoxyphosphinothioyloxyimino)-2-phenylacetonitrile], were studied using buffered sterilized distilled water (pH 4, 7 and 9).     

A new sensitive test based on yeast cells for studying environmental pollution

Different tests based on yeast cells were developed for determination of mutagenic/carcinogenic action; however, they all showed lower sensitivity compared to bacterial tests, the main reason for this being the limited permeability of yeast cells. We found that general permeability of Saccharomyces cerevisiae cells can be increased by mutation and on this basis we developed a more sensitive test. The aim of this study was to prove the applicability of our test, called D7ts1, in environmental studies. Soil, water and air samples were taken during 1998 from regions in Bulgaria with declared low, average or high pollution levels and investigated for presence of mutagenic/carcinogenic activities in the bacterial test of Ames, the yeast D7 test of Zimmermann and our new D7ts1 test. Results obtained evidenced the following conclusions: (1) the usage of D7ts1 test instead of D7 test permits a clearer measurement of positive samples and detects mutagenic/carcinogenic activities undetectable by D7 test; (2) all samples with positive Ames test were positive in the D7ts1 test; however, some samples, clearly positive in the D7ts1, were negative in the Ames test; therefore, the simultaneous usage of D7ts1 and Ames tests in environmental studies is advantageous because it detects dangers for the human health activities to which bacterial cells do not respond; and (3) regions in Bulgaria declared clean were found to be polluted; particularly troubled are the whole-year positive data in the three tests for air samples from a 'clean' region.     

Fugacity modelling to predict the distribution of organic contaminants in the soil:oil matrix of constructed biopiles

Level I and II fugacity approaches were used to model the environmental distribution of benzene, anthracene, phenanthrene, 1-methylphenanthrene and benzo[a]pyrene in a four phase biopile system, accounting for air, water, mineral soil and non-aqueous phase liquid (oil) phase. The non-aqueous phase liquid (NAPL) and soil phases were the dominant partition media for the contaminants in each biopile and the contaminants differed markedly in their individual fugacities. Comparison of three soils with different percentage of organic carbon (% org C) showed that the % org C influenced contaminant partitioning behaviour. While benzene showed an aqueous concentration worthy of note for leachate control during biopiling, other organic chemicals showed that insignificant amount of chemicals leached into the water, greatly reducing the potential extent of groundwater contamination. Level II fugacity model showed that degradation was the dominant removal process except for benzene. In all three biopile systems, the rate of degradation of benzo(a)pyrene was low, requiring more than 12 years for soil concentrations from a spill of about 25 kg (100 mol) to be reduced to a concentration of 0.001 microgg(-1). The removal time of 1-methylphenanthrene and either anthracene or phenanthrene was about 1 and 3 years, respectively. In contrast, benzene showed the highest degradation rate and was removed after 136 days in all biopile systems. Overall, this study confirms the association of risk critical contaminants with the residual saturation in treated soils and reinforces the importance of accounting for the partitioning behaviour of both NAPL and soil phases during the risk assessment of oil-contaminated sites.     

Design and development of novel insect growth regulators: Synthesis,characterization and effect of benzoyl thymyl thioureas and ureas on total haemocyte count of Dysdercus koenigii

Insect-growth regulators (IGRs) have been receiving foremost attention as potential means of selective insect control. Benzoyl phenyl urea (BPU) is a well-known IGR having chitin synthesis inhibitor activity. Mimics of BPU have been synthesized by suitable derivatization of a naturally occurring monoterpenoid, thymol (2-isopropyl-5-methyl phenol) to form a = series of substituted benzoyl thymyl thioureas (BTTUs) [IVa-f] and benzoyl thymyl ureas (BTUs) [Va-f]. The synthesized compounds have been characterized by ¹H and ¹³C NMR, LC-MS and elemental analysis. These derivatives have been screened for their effect on total haemocyte count of Dysdercus koenigii. It has been observed that the introduction of substituted benzoyl thiourea and urea linkage into a thymol ring via an amino group results in higher activity than the parent compound thymol and a comparable pattern of results with the standard insect-growth regulators, Penfluron. Urea [Va-f] compounds exhibited greater effect on Total Haemocyte Count (THC) than thiourea [IVa-f]. Fluoro substitution enhanced the effect on THC more than chloro substituted compounds, while ortho-substitution resulted in a better effect than para-substitution. The results described in this paper are promising and provide new array of synthetic chemicals that may be utilized as insect growth regulators.     

Bioluminescent yeast assays for detecting estrogenic and androgenic activity in different matrices

In this paper we describe the construction and use of a set of bioluminescent yeast strains for the detection of compounds that can affect androgen or estrogen receptor mediated hormonal signalling. The set includes Saccharomyces cerevisiae strains expressing human androgen receptor (AR), estrogen receptor alpha (ERalpha) or estrogen receptor beta (ERbeta), along with firefly luciferase controlled by a respective hormone responsive promoter. A constitutively luminescent strain was included in the set for determining the cytotoxicity of the sample. Yeast cells were incubated with pure chemicals or complex samples for 2.5 h, after which the signal could be detected from the cell-sample mixture after simply adding the D-luciferin substrate. The assays could be completed in one day and they required no cell lysis or centrifugation steps, which makes them suitable for high-throughput analysis of samples. Due to a short incubation time the assays are directly applicable to different sample matrices, requiring no pretreatment of the samples. The assays were used to assess the hormonal activity in moisturizing lotions as an example of a complex sample matrix known to contain endocrine disrupting chemicals. Six out of eight tested moisturisers showed high estrogenic activity, whereas no androgenic activity was observed in the samples.     

Gene expression of heat shock protein 70, interleukin-1β and tumor necrosis factor α as tools to identify immunotoxic effects on Xenopus laevis: a dose-response study with benzo[a]pyrene and its degradation products

The exposure to benzo[a]pyrene (B[a]P) results in an alteration of immune function in mammals and fish, and the analysis of cytokine mRNA levels has been suggested for predicting the immunomodulatory potential of chemicals. To obtain evidence of the innate immune responses to B[a]P in Xenopus laevis, the present study monitored the mRNA expression of interleukin 1-β (IL-1β), tumor necrosis factor α (TNF-α) and heat shock protein 70 (HSP70) in a laboratorial exposure. Tadpoles exposed to 8.36, 14.64, 89.06 and 309.47 μg/L of B[a]P,were used for detecting hsp70, IL-1β and TNF-α mRNA induction. A dose-response increase in the expression of hsp70 and IL-1β mRNA was found. The results of this study confirmed the use of hsp70 and IL-1β, but not TNF-α, as sensitive indicators of immunotoxic effect of B[a]P in X. laevis. Further research would be required for the validation of these endpoints.