Evidence of increased synthesis of δ-aminolevulinic acid dehydratase in experimental lead-poisoned rats

δ-Aminolevulinic acid dehydratase (porphobilinogen synthase; 5-aminolevulinate hydro-lyase, EC 4.2.1.24) was purified from rat and rabbit erythrocytes to a homogeneous state. Specific activities were 26.0 and 26.6 units/mg protein for the rat and rabbit enzymes, respectively, and their estimated molecular weight was 280 000, each consisting of 8 subunits of M_r 35 000. In order to quantitate rat δ-aminolevulinic acid dehydratase at several stages of lead-poisoning, a radioimmunoassay technique using goat antiserum against the rat enzyme was developed for the first time. This technique was specific, reproducible and high sensitive allowing determination of 1 ng enzyme. When drinking water containing 25 mM lead acetate was given daily to rats ad lib. the δ-aminolevulinic acid dehydratase activity in the blood, assayed without any pretreatment, decreased to 8% of the control level on the next day. On the contrary, the restored enzyme activity, assayed in the presence of Zn²⁺ and dithiothreitol, was greater than normal by the fourth day of lead administration in bone-marrow cells and by the ninth day in the peripheral blood. The increased activity level stayed the same from the ninth day onward. The enzyme content as determined directly by the radioimmunoassay technique at this stage was about 2-fold above that the control. There was no significant difference in the number of reticulocytes and the distribution profile of different types of reticulocytes between the lead-exposed and non-exposed rats. Therefore, the increase in the amount of δ-aminolevulinic acid dehydratase in erythrocytes of lead-poisoned rats was suggested to be due to an increased rate of synthesis in the bone-marrow cells.     

Modification of ion transport in lipid bilayer membranes by the insecticides DDT and DDE

In order to elucidate the mechanism of action of organochlorine insecticides on the ion transport in biological membranes, we have studied the effect of DDT and its analog DDE on the structural parameters of phosphatidylethanolamine (PE) planar bilayers. DDT and DDE increase the conductance induced by the hydrophobic ions tetraphenylarsonium (TPhAs⁺) and tetraphenylborate (TPhB^?) in lipid bilayers. Neither DDT nor DDE alters the surface potential of PE monolayers. On the other hand, these organochlorine compounds increase only slightly the electric capacitance of the bilayers. These results are compatible with the hypothesis that these insecticides increase the fluidity of the membrane.     

Levels of Hexachlorocyclohexanes and Dichloro-Dipheny-Trichloroethanes in Penguin Droppings Collected from Ardley Island,Maritime Antarctic

This article reports the concentrations of hexachlorocyclohexanes (HCHs) and dichloro-diphenyl-trichloroethanes (DDTs) in fresh Gentoo penguin (Pygoscelis Papua) droppings deposited on the Ardley Island (62°13′S, 58°56′W), West Antarctica, and those in the island's two lake cores. In the fresh penguin droppings, metabolites were the predominant constituents of the DDT group, most likely due to the ban on DDT use in agriculture and the higher stability of the metabolites in the environment. In contrast, γ-HCH comprised 66% of the total HCHs, apparently due to the continuous use of Lindane in the Southern Hemisphere. The concentrations of HCHs and DDTs in the fresh droppings are many times higher than those in penguin plume, the latter currently being used as biomaterial for monitoring coastal contaminations in Antarctica. Results of this study indicate that penguin droppings have a good potential for use as another biomonitoring material in detecting the organochlorine pesticides and other contaminants in the maritime Antarctic. Because penguin droppings can be well preserved in lake sediments, they also provide a good and continuous historical record of these contaminants. The slight increase of DDTs since the 1990s, as observed in the aged penguin droppings in the sediment cores, suggests there have been new inputs of DDT into the Antarctic coastal ecosystem, probably from the illegal use of DDT in the Southern Hemisphere.     

Application of Semipermeable Membrane Devices (SPMDs) and Benthic Mussels to Evaluate the Bioavailability of Sediment-associated DDTs

Bioavailability of dichlorodiphenyltrichloroethanes (DDTs) in surface sediments was evaluated with semipermeable membrane devices (SPMDs) and two different sediment-dwelling benthic mussels, Bellamya aeruginosa (B. aeruginosa) and Corbicula fluminea (C. fluminea). After 28d laboratory exposure, the positive correlations of DDT concentrations between both SPMDs and benthic mussels with sediments documented that the bioavailability of DDTs was mainly affected by surrounding sediments, while the observed differences of DDT concentrations and congener proportions between B. aeruginosa and C. fluminea were due to the specific physiological characteristics of organisms and different physico-chemical properties of contaminants. Comparisons between SPMDs and benthic mussels showed higher values of biota-sediment accumulation factors (BSAF, 0.63-3.61 for B. aeruginosa and 2.19-17.08 for C. fluminea) than device accumulation factors (DAF, 1.00-1.74). This indicated that living organisms bioaccumulated much more DDTs from sediments than SPMDs due to the different exposure and uptake routes. Strong positive associations between DDTs in SPMDs and benthic mussels indicated SPMDs could mimic the bioaccumulation of DDTs, especially in C. fluminea. However, given the distinct differences observed for both concentrations and congener proportions of DDTs in SPMDs and B. aeruginosa, future study should be directed to develop reliable models with various sediment-dwelling organisms before SPMDs are routinely used in field study.     

Basic studies and applications on bioremediation of DDT: A review

The persistent insecticide DDT (1,1,1-trichloro-2,2-bis (4-chlorophenyl) ethane) has been widely used for pest control in the management of mosquito-borne malaria and is still used for that purpose in some tropical countries. Considering the potential for negative effects due to DDT contamination, it is necessary to determine effective methods of remediation. Several methods have been used to degrade or transform DDT into less toxic compounds. Bacteria and white-rot fungi (WRF) have been shown to enhance the degradation process in soil using both pure and mixed cultures. Recently, a biological approach has been used as an environmentally-friendly treatment, using new biological sources to degrade DDT, e.g. brown-rot fungi (BRF), cattle manure compost (CMC) and spent mushroom waste (SMW). In this review, the abilities of BRF, CMC and SMW to degrade DDT are discussed, including the mechanisms and degradation pathways. Furthermore, application of these sources to contaminated soil is also described. The review discusses which is the best source for bioremediation of DDT.     

Recombinant expression and characterization of Lucilia cuprina CYP6G3: Activity and binding properties toward multiple pesticides

The Australian sheep blowfly, Lucilia cuprina, is a primary cause of sheep flystrike and a major agricultural pest. Cytochrome P450 enzymes have been implicated in the resistance of L. cuprina to several classes of insecticides. In particular, CYP6G3 is a L. cuprina homologue of Drosophila melanogaster CYP6G1, a P450 known to confer multi-pesticide resistance. To investigate the basis of resistance, a bicistronic Escherichia coli expression system was developed to co-express active L. cuprina CYP6G3 and house fly (Musca domestica) P450 reductase. Recombinant CYP6G3 showed activity towards the high-throughput screening substrates, 7-ethoxycoumarin and p-nitroanisole, but not towards p-nitrophenol, coumarin, 7-benzyloxyresorufin, or seven different luciferin derivatives (P450-Glo™ substrates). The addition of house fly cytochrome b₅ enhanced the k_cat for p-nitroanisole dealkylation approximately two fold (17.8 ± 0.5 vs 9.6 ± 0.2 min⁻¹) with little effect on K_M (13 ± 1 vs 10 ± 1 μM). Inhibition studies and difference spectroscopy revealed that the organochlorine compounds, DDT and endosulfan, and the organophosphate pesticides, malathion and chlorfenvinphos, bind to the active site of CYP6G3. All four pesticides showed type I binding spectra with spectral dissociation constants in the micromolar range suggesting that they may be substrates of CYP6G3. While no significant inhibition was seen with the organophosphate, diazinon, or the neonicotinoid, imidacloprid, diazinon showed weak binding in spectral assays, with a Kd value of 23 ± 3 μM CYP6G3 metabolised diazinon to the diazoxon and hydroxydiazinon metabolites and imidacloprid to the 5-hydroxy and olefin metabolites, consistent with a proposed role of CYP6G enzymes in metabolism of phosphorothioate and neonicotinoid insecticides in other species.     

Accelerated internalization of junctional membrane proteins (connexin 43, N-cadherin and ZO-1) within endocytic vacuoles: An early event of DDT carcinogenicity

Stability of cell-to-cell interactions and integrity of junctional membrane proteins are essential for biological processes including cancer prevention. The present study shows that DDT, a non-genomic carcinogen used at a non-cytotoxic dose (1 μM), rapidly disrupted the cell-cell contacts and concomitantly induced the formation of cytoplasmic vacuoles close to the plasma membrane in the SerW3 Sertoli cell line. High-resolution deconvolution microscopy reveals that this vacuolization process was clathrin-dependent since a hyperosmotic media (0.2 M sucrose) blocked rhodamine-dextran endocytosis. In response to DDT, junctional proteins such as Cx43, N-Cadherin and ZO-1 were internalized and present in vacuoles. In Cx43-GFP transfected cells, time lapse videomicroscopy demonstrates that DDT rapidly enhanced fragmentation of the gap junction plaques and abolished the gap junction coupling without major modification of Cx43 phosphorylation status. Repeated exposure to DDT resulted in chronic gap junction coupling injury. The present results demonstrate that one of the early effect of DDT is to interfere with the plasma membrane and to perturb its function, specifically its ability to establish cell-cell junctions that are essential for tissue homeostasis and control of cell proliferation and differentiation. Such an alteration may play a specific role during carcinogenesis.     

Endocrine disruptors (EDs) and hormone-dependent cancers: Correlation or causal relationship?

The selective increase in the incidence of hormone-dependent cancers (breast, prostate, testicular) in industrialized countries is associated with the increasing number of endocrine disruptors (EDs) in the environment and raises questions about the role of EDs in mammary carcinogenesis. Answering these questions is difficult because the number of EDs is large and varies with time. Moreover hormonal carcinogenesis is multifactorial and progresses slowly and in stages. This discussion will be limited to breast cancer and three EDs: distilbene, bisphenol A (BPA), and dichlorodiphenyltrichloroethane (DDT). All these three EDs bind estrogen receptors, albeit with widely different affinities. Several complementary approaches have been used: French cancer records, epidemiological studies on cohorts followed over several decades, numerous in vitro experimental studies using cell cultures and in vivo animal studies. These approaches all converge to the same result, strongly suggesting a causal relationship between EDs and precancerous lesions. Except for distilbene, the mechanisms and molecular targets involved are still unclear, which makes it difficult to look for substitute products that are just as efficient, but less toxic.     

DDT and HCH (Organochlorine Pesticides) in Residential Soils and Health Assessment for Human Populations in Korba,India

Dichlorodiphenyltrichloroethane (DDT), hexachlorocyclohexane (HCH), and their isomers’ levels in residential soils were determined for the assessment of health risk in Korba, India. Observed concentrations of total HCH and total DDT in soils were more or less comparable with other parts of India and the world. ΣHCH and ΣDDT concentrations ranged between 0.9–20 μg kg^?1 and 2–315 μg kg^?1, respectively, which were lower than recommended soil quality guidelines indicating low ecotoxicological risk. Carcinogenic and non-carcinogenic impacts of HCH and DDT on human populations through soil ingestion were evaluated and presented. The incremental lifetime cancer risk (ILCR) for adults and children ranged between 7.8 × 10^?10–1.6 × 10^?7 and 4.1 × 10^?9–8.2 × 10^?7, respectively. Non-cancer health hazard quotient (HQ) ranged between 5.9 × 10^?7–1.8 × 10^?3 and 3.1 × 10^?6–9.4 × 10^?3, respectively, for adults and children. The estimated ILCR and HQ were within the safe acceptable limits of 10^?6–10^?4 and ≤1.0, respectively, indicating low risk to human populations from exposure to organochlorine pesticides (HCH and DDT) in the study area.     

Endophyte-enhanced phytoremediation of DDE-contaminated using Cucurbita pepo: A field trial

Although the use of the pesticide 2,2-bis(p-chlorophenyl)-1,1,1-trichloroethane (DDT) was banned from the mid-1970s, its most abundant and recalcitrant degradation product, 2,2-bis(p-chlorophenyl)-1,1-dichloro-ethylene (DDE), is still present in terrestrial and aquatic ecosystems worldwide.

Zucchini (Cucurbita pepo ssp. pepo) has been shown to accumulate high concentrations of DDE and was proposed for phytoremediation of contaminated soils. We performed a field trial covering a full plant life cycle. C. pepo plants inoculated with the plant growth-promoting endophytic strains Sphingomonas taxi UH1, Methylobacterium radiotolerans UH1, Enterobacter aerogenes UH1, or a consortium combining these 3 strains were grown on a DDE-contaminated field for 100 days. The effects of these inoculations were examined at both the plant level, by evaluating plant weight and plant DDE-content, and at the level of the cultivable and total endophytic communities.

Inoculating plants with S. taxi UH1, M. radiotolerans UH1, and the consortium increased plant weight. No significant effects of the inoculations were observed on DDE-concentrations in plant tissues. However, the amount of DDE accumulated by C. pepo plants per growing season was significantly higher for plants that were inoculated with the consortium of the 3 strains. Therefore, inoculation of C. pepo with DDE-degrading endophytes might be promising for phytoremediation applications.