Interaction of insecticides with lipid membranes

The permeability of liposome membranes is increased by organophosphorus and organochlorinated insecticides at concentrations of 10⁻⁵−10⁻⁴ M. The order of effectiveness is similar to the toxicity of the compounds to mammals, and is the following for permeation of non-electrolytes and for valinomycin-induced permeation of K⁺: parathion > 1,1,1-trichloro-2,2-bis(p-chlorophenyl) ethane (DDT) ≈ aldrin malathion > lindane. The degree of effectiveness for X-537A-induced permeation of Ca²⁺ was the following: aldrin DDT > parathion malathion > lindane. The organophosphorus compound, ethyl azinphos (10⁻⁴ M), dramatically increases the permeability of liposome membranes to all the tested substances, probably as a consequence of surfactant effects. Some organochlorinated insecticides appear to react with cation ionophores and modulate their motion across lipid membranes.It is suggested that the insecticides may exert some of their toxic actions by modifying certain mechanisms in the cell membrane.     

Bacterial bioconcentration of chlorinated hydrocarbon insecticides from aqueous systems

The prevalence of chlorinated hydrocarbon insecticide uptake by chemoorganotrophic bacteria has been investigated. Thirteen bacterial species were observed to sorb and concentrate (bioconcentratc)α-chlordane,β-chlordane, dieldrin, heptachlor epoxide, and lindane from aqueous systems. Bioconcentration, as expressed by the ratio of cellular insecticide in ng/mg (dry wt) to supernatant insecticide in ng/μl, ranged from 10 for lindane byEnterobacter aerogenes to a high of 55,900 forβ-chlordane byCaulobacter vibrioides var.limonus. Amounts of cellular chlorinated hydrocarbon insecticides (CHI) detected and the bioconcentration ratios were observed to have the following order in magnitude:α- orβ-chlordane > dieldrin > heptachlor epoxide > lindane. This decreasing order was the inverse of reported water solubilities for the CHI and the inverse relationship was mathematically defined. The CHI were not easily removed from cells by washing (desorbing) and desorption was directly proportional to insecticide water solubility. Uptake of the CHI was rapid, near-maximum amounts being sorbed within 15 min, and pH 7 appeared optimal for bioconcentration as examined over the range pH 6 to 8. Implications of this investigation are that bioconcentration of CHI by bacteria might serve as a means of introducing these toxic compounds into aquatic food chains and that the bioconcentration phenomenon might lend itself as a treatment procedure for the intentional removal of residual CHI from water supplies and wastewater.     

Residues of chlorinated hydrocarbon insecticides in the North Sea environment

Summary 1. Sandwich terns(Sterna sandvicensis) of a colony in the Dutch Wadden Sea, which were seen dying in tremors and convulsions, were analysed for their contents of chlorinated hydrocarbon insecticides. These proved to be present in considerable concentrations.2. The concentrations found were compared to those present in chickens of hens, poisoned in the laboratory. The amounts present in the dying or dead Sandwich terns were high enough to cause their death.3. Tern eggs from different coastal places in Ireland, Great Britain, The Netherlands and West Germany were sampled and analysed for these compounds. The amounts of dieldrin, telodrin and endrin in the eggs from the first two countries were much lower than those in the eggs from the last two ones. The amounts of DDE were quite the same.4. Tern food fishes(Clupea sprattus, Clupea harengus andAmmodytes lanceolatus) were sampled in the Dutch Wadden Sea and analysed. They contained considerable amounts of chlorinated hydrocarbon insecticides.5. Mussels(Mytilus edulis) were sampled at a series of locations alongside the Dutch coast. The highest concentrations of the insecticides were found on places near the mouth of the river Rhine. Also telodrin, an insecticide not used in Europe, was found.6. A big insecticide producing industry near the mouth of the river Rhine was informed of these facts. Measures taken by this form resulted in a decrease of the amounts of telodrin in the coastal North Sea environment.

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Rückstände von chlorierten Kohlenwasserstoff-Insektiziden im Lebensraum der Nordsee

Kurzfassung In einer großen Brutkolonie von Brandseeschwalben(Sterna sandvicensis) auf der Insel Griend im holländischen Wattenmeer war die Anzahl der Brutvögel von > 20 000 Paaren im Jahre 1955 auf etwa 1000 Paare im Jahre 1965 zurückgegangen. Viele Vögel wurden tot oder sterbend mit Krampfsymptomen aufgefunden. Organe und Blut dieser Vögel wurden auf ihren Gehalt an nerventoxischen Stoffen, insbesondere an chlorierten Kohlenwasserstoff-Insektiziden, untersucht. Die Konzentrationen wurden mit denen von Jungvögeln verglichen, die im Laboratorium auf künstliche Weise mit diesen Stoffen versehen worden waren und daran auch starben. Es konnte somit nachgewiesen werden, daß die in den Brandseeschwalben gefundenen Konzentrationen zum Tod der Tiere geführt haben. Die an verschiedenen Orten gesammelten Eier der Brandseeschwalben wurden ebenfalls untersucht. Eier aus den Niederlanden und aus Deutschland enthielten deutlich mehr Endrin, Dieldrin und Telodrin als Eier aus England und Irland. Verschiedene Fischarten(Clupea sprattus, Clupea harengus, Ammodytes lanceolatus), die von den Brandseeschwalben gefressen werden, wurden im holländischen Wattenmeer gesammelt und analysiert; sie enthielten beträchtliche Mengen an chlorierten Kohlenwasserstoff-Insektiziden. Der Herkunft dieser Stoffe wurde durch Untersuchungen von Miesmuscheln(Mytilus edulis) nachgegangen, die an verschiedenen Stellen entlang der holländischen Küsten gesammelt wurden. Die größten Insektizid-Konzentrationen wurden in Miesmuscheln gefunden, die aus dem Gebiet der Rheinmündung stammten. Einem dort gelegenen chemischen Werk, das diese Insektenvertilgungsmittel herstellt, wurden die Untersuchungsergebnisse mitgeteilt. Zugleich wurden geeignete Maßnahmen vorgeschlagen, um das Einleiten von insektizidhaltigen Abwässern zu verhindern. Augenscheinlich hat inzwischen die Konzentration einiger dieser Stoffe inMytilus edulis abgenommen.

     

Partition of lindane in synthetic and native membranes

Partition coefficients of the insecticide γ-1,2,3,4,5,6-hexachlorocyclohexane (trivially, lindane) were determined in model and native membranes. Partition in egg phosphatidylcholine bilayers decreases linearly with temperature, over a range (10–40°C) at which the lipid is in the liquid-crystalline state. Addition of 50 mol% cholesterol dramatically decreases partition (2100 falls to 100, at 10°C) and abolishes the temperature dependence. First-order phase transitions of dimyristoyl-, dipalmitoyl- and distearoylphosphatidylcholines (DMPC, DPPC and DSPC) are accompanied by a sharp increase in lindane partition. Apparently, the insecticide is easily accommodated in bilayers of short-aliphatic-chain lipids, since the partitions were 2450, 600 and 50 in DMPC, DPPC and DSPC, respectively, at temperatures 10 Cdeg below the midpoint of their transitions. The lindane partition sequence in native membranes is as follows: mitochondria, sarcoplasmic reticulum, myelin, brain microsomes and erythrocytes. This sequence correlates reasonably well with the relative content of cholesterol and is similar in liposomes of total extracted lipids, although the absolute partitions showed decreased values. Therefore, the presence of proteins in native membranes contributes to the insecticide partition, probably by favouring its interaction with lipids.     

Decline of chlorinated hydrocarbon insecticides residues in the tea-garden soils of Taiwan

Residues of chlorinated hydrocarbon insectidies and some of their metabolites, including p,p'-DDT, p,p'-DDD, p,p'-DDE, gamma-BHC, aldrin, dieldrin, heptachlor and heptachlor epoxide, were examined in the soils collected from 21 tea-gardens distributed in 15 countries of 6 prefectures in 1977, and from 100 locations distributed in 46 counties of 10 prefectures in 1984 in Taiwan. Although the use of these chlorinated hydrocarbon insecticides has been banned for many years, minor amounts of residues in the soils have still been detected. From 1977 to 1984, average residues of the chlorinated hydrocarbon insecticides and their metabolites in the soils of tea-gardens decreased from 5.64 to 0.06 ppb for gamma-BHC, 97 54 to 1.10 ppb for aldrin, 35.91 to 1.00 ppb for p,p'-DDE, 17.41 to 0.28 ppb for dieldrin, and 30.17 to 3.42 ppb for p,p'-DDT. But the number of positive cases increased for all chlorinated hydrocarbon insecticides tested, the residual levels were not significantly different among soils with different physiochemical properties or among soil layers. No significant amount of these insecticides was found in the processed tea.     

Optically active gossypol as a circular dichroism probe of interactions with serum albumins

The (+)-enantiomer of the polyphenolic binaphthyl gossypol, has been shown to be a useful CD probe of interactions with human and bovine serum albumin. (+)-Gossypol binds to albumin with same affinity as recemic (±)-gossypol, as shown by fluorescence quenching, and also displaces bilirubin from its albumin binding site. The CD characteristics of bound gossypol are different in the case of the two proteins.     

Interactions of chlorinated hydrocarbon pesticides with the 8S estrogen-binding protein in rat testes

The effect of certain DDT analogs on the binding of ³H-estradiol to the 8–9S estrogen binding protein of rat testicular cytosol was studied by sucrose sedimentation analysis. The binding of ³H-estradiol to testicular cytosol was inhibited by o,p'DDT, a DDT analog which is estrogenic in the intact female, but not by p,p'DDE which is a nonestrogen in the female. The pesticide methoxychlor, which is estrogenic $\text{in vivo}$ in the female, failed to inhibit ³H-estradiol binding, presumably requiring metabolic activation for binding to the testicular cytosol. In fact, its di-demethylated metabolite 2,2-bis(p-hydroxyphenyl)-1, 1,1-trichloroethane (HPTE), also estrogenic $\text{in vivo}$, caused $\text{marked}$ suppression of ³H-estradiol binding.     

Interaction of insecticides with mammalian P-glycoprotein and their effect on its transport function

We studied the effects of four commonly used insecticides (methylparathion, endosulfan, cypermethrin and fenvalerate) on P-glycoprotein isolated from multidrug-resistant cells. All the pesticides stimulated P-glycoprotein ATPase activity, with maximum stimulation of up to 213% in a detergent-solubilized preparation, and up to 227% in reconstituted liposomes. The ATPase stimulation profiles were biphasic, displaying lower stimulation, and in the case of methylparathion, inhibition of activity, at higher insecticide concentrations. Quenching of the intrinsic Trp fluorescence of purified P-glycoprotein was used to quantitate insecticide binding; the estimated K_d values fell in the range 4-6 μM. Transport of the fluorescent substrate tetramethylrosamine (TMR) into proteoliposomes containing P-glycoprotein was monitored in real time. The TMR concentration gradient generated by the transporter was collapsed by the addition of insecticides, and prior addition of these compounds prevented its formation. The rate of TMR transport was inhibited in a saturable fashion by all the compounds, indicating that they compete with the substrate for membrane translocation. Taken together, these data suggest that the insecticides bind to Pgp with high affinity and effectively block drug transport. Inhibition of Pgp by pesticides may compromise its ability to clear xenobiotics from the body, leading to a higher risk of toxicity.     

Reaction of the soil microflora after contamination with chlorinated aromatic compounds and HCH

Abstract The effect of the pollution of an industrial land site with chlorinated benzenes, chlorinated phenols, hexachlorocyclohexane-isomers (HCH) on the soil microflora was investigated. Cell counts (microscopic and by plate count) as well as respiration rated did not correlate negatively with the concentration of the contaminants. Soil microorganisms grew in the presence of up to 750 μmol 1⁻¹ pf chlorinated compounds in liquid culture. Only 150 μmol l⁻¹ 2,4,5-trichlorophenol (2,4,5-TCP) inhibited growth totally. In enrichment cultures, bacteria used α- and γ-HCH, 3-chlorophenol (3-CP), 2,3-dichlorophenol (2,3-DCP), 2,6-DCP, 2,4,5-TCP, and 1,2,4,5-tetrachlorobenzene (1,2,4,5-TeCB) as a sole source of carbon and energy under aerobic conditions. No growth was observed with β-HCH. Under anaerobic conditions no growth was observed with any of the substances tested     

Depth profiling in membranes by fluorescence quenching

Membrane penetration depth is an important parameter in relation to membrane structure and organization. A methodology has been developed to analyze the membrane penetration depths of fluorescent molecules or groups utilizing differential fluorescence quenching caused by membrane embedded spin-label probes located at different depths. The method involves determination of the parallax in the apparent location of fluorophores, detected when quenching by phospholipids spin-labelled at two different depths is compared. By use of relatively simple algebraic expressions, the method allows calculation of depth in å. This method has been used to determine the location of fluorophores in NBD-labelled lipids and anthroyloxy-labelled fatty acids in model membranes and of the membrane embedded tryptophan residues in the reconstituted nicotinic acetylcholine receptor.     

The influence of some chlorinated hydrocarbon insecticides on the mineralization of N fertilizers and plant growth

Summary The variable effects, previously observed under field and laboratory conditions, resulting from the application of the widely used chlorinated hydrocarbon insecticides has stimulated further investigations. Because of the possible cumulative harmful effects of the insecticides on the activities of soil micro-organisms, several investigations have been concerned with the influence of these insecticides on the conversion of ammonia to nitrate.The present investigation is concerned with the relationship between the deleterious effect of certain pesticides on the activities of the soil microflora and the use of nitrogenous fertilizers; an aspect of the problem which has not previously been considered.The results of the investigation may allow the anomalous effects of the insecticides in the laboratory and in the field to be understood.The observations indicate that the basis of the effects of hexachlorocyclohexane on soils is as an inhibitor of nitrification.     

Interactions of metal ions with phosphatidylserine bilayer membranes: effect of hydrocarbon chain unsaturation

A combination of surface monolayer, scanning calorimetry, 31P NMR, and spin-label ESR techniques has been used to monitor the interactions of monovalent (NH4+, Na+, and Li+) and divalent (Ca2+) cations with phosphatidylserines (PS) differing in their levels of chain unsaturation. Comparisons are made between the disaturated dimyristoyl-, dipalmitoyl-, and dihexadecyl-PS (DMPS, DPPS, and DHPS), saturated cis-monounsaturated palmitoyloleoyl-PS (POPS) (and bovine brain PS), di-trans-monounsaturated dielaidoyl-PS (DEPS), and di-cis-monounsaturated dioleoyl-PS (DOPS). Na+ and NH4+ cations interact weakly with all PS monolayers and bilayers without significant changes in molecular conformation, chain packing, or headgroup dynamics and without dependence on chain composition. In contrast, considering these structural and dynamic parameters, Li+ shows a gradation in its interaction with PS (DMPS greater than POPS approximately bovine brain PS greater than DOPS), suggesting that Li+-PS interactions depend on the interfacial properties of the PS molecules (e.g., surface area). Finally, Ca2+ interacts strongly with all PS monolayers and bilayers, without obvious chain selectivity. Thus, ion binding to PS depends not only on the properties of the cation (Na+ vs Li+ vs Ca2+) but also on the molecular details of the PS membrane surface.     

Microbial transformation of chlorinated benzoates

Chlorinated benzoates enter the environment through their use as herbicides or as metabolites of other halogenated compounds. Ample evidence is available indicating biodegradation of chlorinated benzoates to CO₂ and chloride in the environment under aerobic as well as anaerobic conditions. Under aerobic conditions, lower chlorinated benzoates can serve as sole electron and carbon sources supporting growth of a large list of taxonomically diverse bacterial strains. These bacteria utilize a variety of pathways ranging from those involving an initial degradative attack by dioxygenases to those initiated by hydrolytic dehalogenases. In addition to monochlorinated benzoates, several bacterial strains have been isolated that can grow on dichloro-, and trichloro- isomers of chlorobenzoates. Some aerobic bacteria are capable of cometabolizing chlorinated benzoates with simple primary substrates such as benzoate. Under anaerobic conditions, chlorinated benzoates are subject to reductive dechlorination when suitable electron-donating substrates are available. Several halorespiring bacteria are known which can use chlorobenzoates as electron acceptors to support growth. For example, Desulfomonile tiedjei catalyzes the reductive dechlorination of 3-chlorobenzoate to benzoate. The benzoate skeleton is mineralized by other microorganisms in the anaerobic environment. Various dichloro- and trichlorobenzoates are also known to be dechlorinated in anaerobic sediments.     

Three new chlorinated phenolic glycosides from Przewalskia tangutica

Three new chlorinated phenolic glycosides, namely przewatangosides A-C (1-3), along with one known compound, globosumoside A (4), were isolated from the whole plants of Przewalskia tangutica. Their structures were unequivocally determined by extensive spectroscopic analysis and chemical method. The cytotoxic activities of the isolated phenolic glycosides (1-4) were evaluated against the five human cancer cell lines A549, MCF-7, SMMC-7721, HepG2 and HL-60. Przewatangoside A (1) exhibited weak cytotoxicity against SMMC-7721 with the IC₅₀ value of 38.1 μM. All the tested compounds were inactive (IC₅₀ > 50 μM) to the normal human hepatocyte cell line (L02).     

Interactions of Chromium (III) and Chromium (VI) with Bovine Serum Albumin Studied by UV Spectroscopy,Circular Dichroism,and Fluorimetry

In the present work, the interactions of bovine serum albumin (BSA) with chromium (III) chloride, potassium dichromate, and chromate were studied by fluorescence, circular dichroism, and UV–vis absorbance spectroscopy. Fluorescence quenching of BSA by chromium (III) was found to be a dynamic process in the beginning, turning static at later stages. Spectroscopic data show that both dichromate and chromate bind in similar electrostatic fashion to BSA and does not follow the fluorescence quenching observation for chromium (III).