Hepatoprotective activity of Rhus oxyacantha root cortex extract against DDT-induced liver injury in rats.

The present investigation aimed to study the antioxidant activity and hepatoprotective effects of ethyl acetate extract of R. oxyacantha root cortex (RE) against DDT-induced liver injury in male rats. The RE exhibited high total phenolic, flavonoid and condensed tannins contents. The antioxidant activity in vitro systems showed a significant potent free radical scavenging activity of the extract. The HPLC finger print of R. oxyacantha active extract showed the presence of five phenolic compounds with higher amounts of catechol and gallic acid. The in vivo results showed that a single intraperitoneal administration of DDT enhanced levels of hepatic markers (ALT, AST and LDH) in serum of experimental animals. It also increased the oxidative stress markers resulting in increased levels of the lipid peroxidation with a significant induction of SOD and GPx, metallothioneins (MTs) and a concomitant decrease of non protein thiols (NPSH) in liver. However, pretreatment of rats with RE at a dose of 150 and 300mg/kg body weight significantly lowered serum transaminases and LDH in treated rats. A significant reduction in hepatic thiobarbituric reactive substances and a decrease in antioxidant enzymes activities and hepatic MTs levels by treatment with plant extract against DDT, were observed. These biochemical changes were consistent with histopathological observations, suggesting marked hepatoprotective effect of RE with the two doses used. These results strongly suggest that treatment with ethyl acetate extract normalizes various biochemical parameters and protects the liver against DDT-induced oxidative damage in rats and thus help in evaluation of traditional claim on this plant.

Insecticidal effect of plant extracts on Phlebotomus argentipes (Diptera: Psychodidae) in Bihar, India.

BACKGROUND & OBJECTIVES: Phlebotomus argentipes (Diptera: Psychodidae), the established vector for kala-azar is presently being controlled by indoor residual spray of DDT in kala-azar endemic areas in India. Search for non-hazardous and non-toxic biodegradable active molecules from botanicals may provide cost-effective and eco-friendly alternatives to synthetic insecticides. The present study was aimed at evaluating various plant extracts from endemic and non-endemic areas of Bihar for their insecticidal activity against sandfly to identify the most effective plant extract. METHODS: Bio-assay test was conducted with larvae and adult of P. argentipes with different plant extracts collected in distilled water, hexane, ethyl acetate, acetone and methanol. Thin layer chromatography (TLC), column chromatography and high performance liquid chromatography (HPLC) were conducted for detection of active molecules. RESULTS: Adults and larvae of sandflies exposed to the aqueous extract of Nicotiana tabacum resulted in 100 per cent mortality. The hexane extract of Clerodendrum infortunatum was found to kill 77 per cent adults but was ineffective against larvae. Bio-assay test of the ninth fraction (hexane extract-methanol phase) separated by column chromatography was found to be 63 per cent effective. The purple spot on the TLC of this fraction indicated the presence of a diterpenoid. HPLC of this fraction detected nine compounds with two peaks covering 20.44 and 56.52 per cent areas with retention time of 2.439 and 5.182 min, respectively supporting the TLC results. INTERPRETATION & CONCLUSIONS: The column separated 9 [th] fraction of C. infortunatum extract was found to be effective in killing 63 per cent of adult P. argentipes. Compounds of this fraction need to be evaluated further for identification and characterization of the active molecule by conducting individual bio-assay tests followed by further fractionation and HPLC. Once the structure of the active molecule is identified and validated, it may be synthesized and formulated as a product.

Effects of formoterol on protein metabolism in myotubes during hyperthermia.

Proteolysis in skeletal muscle is mainly carried out by the activity of the ubiquitin-dependent proteolytic system. For the study of protein degradation through the ubiquitin-proteasome pathway, we used a model of hyperthermia in murine myotubes. In C2C12 cells, hyperthermia (41°C) induced a significant increase in both the rate of protein synthesis (18%) and degradation (51%). Interestingly, the addition of the ß(2) -adrenoceptor agonist formoterol resulted in a significant decrease in protein degradation (21%) without affecting protein synthesis. The decrease in proteolytic rate was associated with decreases in gene expression of the different components of the ubiquitin-dependent proteolytic system. The effects of the ß(2) -agonist on protein degradation were dependent exclusively on cAMP formation, because inhibition of adenylyl cyclase completely abolished the effects of formoterol on protein degradation. It can be concluded that hyperthermia is a suitable model for studying the anti-proteolytic potential of drugs used in the treatment of muscle wasting.

Effects of selected food phytochemicals in reducing the toxic actions of TCDD and p,p'-DDT in U937 macrophages.

To assess the effectiveness of selected food phytochemicals in reducing the toxic effects of the environmental toxicants, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and p,p'-DDT (DDT), we tested the potencies of auraptene, nobiletin, zerumbone, and (±)-13-hydroxy-10-oxo-trans-11-octadecenoic acid (13-HOA) in reversing the inflammatory action of these toxicants in U937 human macrophages. Using quantitative RT-PCR as the initial screening assay, we identified antagonistic actions of zerumbone and auraptene against the action of TCDD and DDT in up-regulating the mRNA expressions of COX-2 and VEGF. The functional significance of the inhibitory action of zerumbone on COX-2 expression was confirmed by demonstrating its suppression of TCDD-induced activation of COX-2 gene expression in mouse MMDD1 cells. We tested auraptene on DDT-induced reactive oxygen species (ROS) formation in U937 macrophages and found that auraptene is a powerful agent antagonizing this action of DDT. To confirm the significance of these actions of zerumbone and auraptene at the cellular level, we assessed their influence on TCDD-induced apoptosis resistance in intact U937 macrophages and found that they are capable of reversing this action of TCDD. In conclusion, zerumbone and auraptene were identified to be the most effective agents in protecting U937 macrophages from developing these cell toxic effects of TCDD and DDT.

Program to eradicate malaria in Sardinia, 1946-1950.

During 1946-1950, the Rockefeller Foundation conducted a large-scale experiment in Sardinia to test the feasibility of indigenous vector species eradication. The interruption of malaria transmission did not require vector eradication, but with a goal of developing a new strategy to fight malaria, the choice was made to wage a rapid attack with a powerful new chemical. Costing millions of dollars, 267 metric tons of DDT were spread over the island. Although malaria was eliminated, the main objective, complete eradication of the vector, was not achieved. Despite its being considered almost eradicated in the mid-1940s, malaria 60 years later is still a major public health problem throughout the world, and its eradication is back on the global health agenda.

Presence of a putative steroidal allosteric site on glycoprotein hormone receptors.

In a previous work we found that the insecticide 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT), inhibits the accumulation of cAMP as induced by the bovine thyroid stimulating hormone (bTSH) in cells transfected with the TSH receptor. In this work, we demonstrate that the DDT molecular analogues, diethylstilbestrol and quercetine, are more potent inhibitors of the TSH receptor activity than DDT itself. The notion that all these compounds interfere with nuclear estrogen receptors, as either agonists (DDT and diethylstilbestrol) or antagonists (quercetin), prompted us to test the ability of the steroid hormone 17-beta-estradiol to inhibit the TSH receptor activity. We found that estrogen exposure causes a modest but significant inhibition of the bTSH induced cAMP accumulation both in transfected CHO-TSH receptor and Fischer Rat Thyroid Low Serum 5% (FRTL-5) cells. When applied to CHO cells transfected with the luteinizing hormone receptor, 17-beta-estradiol proved capable of inhibiting the hCG induced cAMP accumulation at a concentration as low as 10nM, though the effect was not greater than 35%. The effect of 17-beta-estradiol was not estrogen receptors mediated, as co-transfection of the estrogen receptor alpha and beta subunits with LH receptor caused cAMP to increase above the level attained by the sole hCG stimulation, and not to decrease it as expected. These data suggest the presence of a steroidal-like allosteric binding site on glycoprotein hormone receptors.

Gene expression profiling in Ishikawa cells: a fingerprint for estrogen active compounds.

Several anthropogenous and naturally occurring substances, referred to as estrogen active compounds (EACs), are able to interfere with hormone and in particular estrogen receptor signaling. EACs can either cause adverse health effects in humans and wildlife populations or have beneficial effects on estrogen-dependent diseases. The aim of this study was to examine global gene expression profiles in estrogen receptor (ER)-proficient Ishikawa plus and ER-deficient Ishikawa minus endometrial cancer cells treated with selected well-known EACs (diethylstilbestrol, genistein, zearalenone, resveratrol, bisphenol A and o,p'-DDT). We also investigated the effect of the pure antiestrogen ICI 182,780 (ICI) on the expression patterns caused by these compounds. Transcript levels were quantified 24 h after compound treatment using Illumina BeadChip Arrays. We identified 87 genes with similar expression changes in response to all EAC treatments in Ishikawa plus. ICI lowered the magnitude or reversed the expression of these genes, indicating ER dependent regulation. Apart from estrogenic gene regulation, bisphenol A, o,p'-DDT, zearalenone, genistein and resveratrol displayed similarities to ICI in their expression patterns, suggesting mixed estrogenic/antiestrogenic properties. In particular, the predominant antiestrogenic expression response of resveratrol could be clearly distinguished from the other test compounds, indicating a distinct mechanism of action. Divergent gene expression patterns of the phytoestrogens, as well as weaker estrogenic gene expression regulation determined for the anthropogenous chemicals bisphenol A and o,p'-DDT, warrants a careful assessment of potential detrimental and/or beneficial effects of EACs. The characteristic expression fingerprints and the identified subset of putative marker genes can be used for screening chemicals with an unknown mode of action and for predicting their potential to exert endocrine disrupting effects.

Stimulation of transactivation of the largemouth bass estrogen receptors alpha, beta-a, and beta-b by methoxychlor and its mono- and bis-demethylated metabolites in HepG2 cells.

The purpose of this study was to determine the mechanisms by which the pesticide, methoxychlor (MXC), acts as an environmental endocrine disruptor through interaction with the three largemouth bass (Micropterus salmoides) estrogen receptors (ERs) alpha, betaa, and betab. MXC is a less-environmentally persistent analog of DDT that behaves as a weak estrogen. Using transient transfection assays in HepG2 cells, we have previously shown that each receptor is responsive to the endogenous ligand 17beta-estradiol (E(2)) in a dose-dependent manner. The parent compound, MXC, showed dose-dependent stimulation of transcriptional activation through all three ERs. In addition to the parent molecule, each of the metabolites was also estrogenic with all three ERs. The order of potency for ERalpha and ERbetab was HPTE>OH-MXC>MXC, while the opposite order was seen for ERbetaa. HepG2 cells did not substantially metabolize MXC to the active metabolites, thus the activity of MXC was not due to metabolism. When examining the effects of increasing concentrations of MXC at a fixed concentration of E(2), all three ERs show increased activity compared to that with E(2) alone, showing that the effects of MXC and E(2) are additive. However, when this experiment was repeated with increasing concentrations of HPTE at a fixed concentration of E(2), the activity of ERalpha was decreased, that of ERbetab was increased, while that of ERbetaa was unaffected compared to E(2) alone. These experiments suggest that HPTE functions as an E(2) antagonist with ERalpha, an E(2) agonist with ERbetab and does not perturb E(2) stimulation of ERbetaa. While it is clear the ERbeta subtypes are the products of different genes (due to a gene duplication in teleosts) the differences in their responses to MXC and its metabolites indicate that their functions diverge, both in their in vivo molecular response to E(2), as well as in their interaction with endocrine disrupting compounds found in the wild.

Flow cytometric cell cycle analysis allows for rapid screening of estrogenicity in MCF-7 breast cancer cells.

The quantitative measurement of individual cells and their characteristics by means of flow cytometry is already for many years of great value for clinical studies. However, its potential as a tool in (eco)toxicology has only recently been discovered. Analysis of cell cycle kinetics with DNA-staining dyes can offer a valuable alternative to detect effects of chemicals on cell proliferation, an important endpoint in screening estrogen-like properties of chemicals. In the present study, flow cytometric cell cycle analysis in growth arrested MCF-7 cells exposed to five xenoestrogens correspond well with cell proliferation results of the conventionally used E-screen assay. Moreover, re-induction of proliferation in MCF-7 cells, indicated by the percentage of cells in S(ynthesis)-phase, is most pronounced after 24 h exposure, thus allowing a faster screening of xenoestrogens. This flow cytometric proliferation assay confirms that the estrogenic activity of structurally analogous parabens is mediated by the estrogen receptor pathway and is proportional to the alkyl chain length. Moreover, the ER-mediated mode of action of two fluorotelomer alcohols (6:2 FTOH and 8:2 FTOH), recently reported as xenoestrogenic, could be elucidated. These results support the potential of flow cytometric cell cycle kinetics as a screening assay for estrogen-like properties of chemicals.

Analogous pleiotropic effects of insecticide resistance genotypes in peach-potato aphids and houseflies.

We show that single-point mutations conferring target-site resistance (kdr) to pyrethroids and DDT in aphids and houseflies, and gene amplification conferring metabolic resistance (carboxylesterase) to organophosphates and carbamates in aphids, can have deleterious pleiotropic effects on fitness. Behavioural studies on peach-potato aphids showed that a reduced response to alarm pheromone was associated with both gene amplification and the kdr target-site mutation. In this species, gene amplification was also associated with a decreased propensity to move from senescing leaves to fresh leaves at low temperature. Housefly genotypes possessing the identical kdr mutation were also shown to exhibit behavioural differences in comparison with susceptible insects. In this species, resistant individuals showed no positional preference along a temperature gradient while susceptible genotypes exhibited a strong preference for warmer temperatures.

Ability of xeno- and phytoestrogens to modulate expression of estrogen-sensitive genes in rat uterus: estrogenicity profiles and uterotropic activity.

The function of the uterus is regulated by female sex steroids and it is, therefore, used as the classical target organ to detect estrogenic action. Uterine response to estrogens involves the activation of a large pattern of estrogen-sensitive genes. This fact offers the opportunity to analyze the estrogenic activity of xeno- and phytoestrogens, and the mechanisms of their molecular action by a correlation of the uterotropic activity and their ability to modulate the expression of estrogen-sensitive genes. We have analyzed the expression of androgen receptor (AR), progesterone receptor (PR), estrogen receptor (ER), clusterin (CLU), complement C3 (C3), and GAPDH mRNA in the rat uterus following oral administration of ethinylestradiol (EE), bisphenol A (BPA), o,p'-DDT (DDT), p-tert-octylphenol (OCT) and daidzein (DAI). A significant stimulation of the uterine wet weight could be observed after administration of all the substances. The activity of all analyzed compounds to stimulate uterine weight was low in comparison to EE. DDT has the highest activity to stimulate uterine weight whereas BPA and DAI turned out to be less potent. The analysis of gene expression revealed a very specific profile of molecular action in response to the different compounds which cannot be detected by judging the uterotropic response alone. A dose dependent analysis revealed that C3 mRNA is already modulated at doses where no uterotropic response was detectable. Although DAI and BPA were very weak stimulators of uterine growth, these substances were able to alter the expression of AR, ER and C3 very strongly. Based on these investigations the analyzed compounds can be subdivided into distinct classes: First, compounds which exhibit a similar gene expression fingerprint as EE (e.g. OCT); second, compounds exhibiting a significant uterotropic activity, but inducing a pattern of gene expression different from EE (e.g. DDT); and third, compounds like BPA and especially DAI which exhibit a very low uterotropic activity, but nevertheless modulate the expression of estrogen-sensitive genes. These findings strongly suggest that the fingerprint of uterine gene expression is a very sensitive tool to investigate estrogenicity of natural and synthetic compounds and offers the possibility to get information in regard to the molecular mechanisms involved in the action of the respective compounds.

Determining relative estrogenicity by quantifying vitellogenin induction in rainbow trout liver slices.

Precision cut tissue slices, by modeling the entire organ, are a valuable tool for studying protein induction or inhibition by test chemicals. This manuscript describes parameters to quantify relative estrogenicity of chemicals in rainbow trout liver slices by measuring vitellogenin (Vg) induction, a well-characterized biomarker of estrogen receptor signal transduction. Hank's medium (phenol-red free) supplemented with Hepes, sodium bicarbonate, and 1% bovine serum albumin was utilized. The experimental parameters were optimized using 1000 nM 17beta-estradiol, a potent estrogen in rainbow trout that induces Vg production in vivo. The addition of trout serum and retention of the media was essential, probably to allow for the accumulation of Vg in the slices and media. Histological examination and ATP analyses indicated no toxicity in control or 17beta-estradiol-treated liver slices after 120 h. Induction was 4-fold greater with 25% serum containing media compared to media with 10% serum. We observed Vg induction as great as 500-fold over controls at 96 h in liver slices and media containing 25% serum and 1000 nM 17beta-estradiol. Controls without 17beta-estradiol, incubated in media with 10 or 25% serum, exhibited no detectable Vg production, indicating that the induction seen was not from the media or serum. We observed that 48 h was required for significant Vg induction in the media and liver slices. Maximum induction in slices occurred at 96 h, whereas media Vg levels continued to increase to 120 h, suggesting a time delay between Vg production and excretion by the liver. The feasibility of this model to detect weak environmental estrogens was determined with 0-250 microM o,p'DDE and bisphenol A. Both compounds induced Vg in this model with EC50 values of 10(4) and 2x10(5) higher than E(2), respectively. Our results indicate the importance of media, serum, and time selection for optimal Vg induction. This model allows for the determination of relative estrogenicity of chemicals in a controlled in vitro system while utilizing the advantages of precision cut slice technology.

Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor beta.

The rat, mouse and human estrogen receptor (ER) exists as two subtypes, ER alpha and ER beta, which differ in the C-terminal ligand-binding domain and in the N-terminal transactivation domain. In this study, we investigated the estrogenic activity of environmental chemicals and phytoestrogens in competition binding assays with ER alpha or ER beta protein, and in a transient gene expression assay using cells in which an acute estrogenic response is created by cotransfecting cultures with recombinant human ER alpha or ER beta complementary DNA (cDNA) in the presence of an estrogen-dependent reporter plasmid. Saturation ligand-binding analysis of human ER alpha and ER beta protein revealed a single binding component for [3H]-17beta-estradiol (E2) with high affinity [dissociation constant (Kd) = 0.05 - 0.1 nM]. All environmental estrogenic chemicals [polychlorinated hydroxybiphenyls, dichlorodiphenyltrichloroethane (DDT) and derivatives, alkylphenols, bisphenol A, methoxychlor and chlordecone] compete with E2 for binding to both ER subtypes with a similar preference and degree. In most instances the relative binding affinities (RBA) are at least 1000-fold lower than that of E2. Some phytoestrogens such as coumestrol, genistein, apigenin, naringenin, and kaempferol compete stronger with E2 for binding to ER beta than to ER alpha. Estrogenic chemicals, as for instance nonylphenol, bisphenol A, o, p'-DDT and 2',4',6'-trichloro-4-biphenylol stimulate the transcriptional activity of ER alpha and ER beta at concentrations of 100-1000 nM. Phytoestrogens, including genistein, coumestrol and zearalenone stimulate the transcriptional activity of both ER subtypes at concentrations of 1-10 nM. The ranking of the estrogenic potency of phytoestrogens for both ER subtypes in the transactivation assay is different; that is, E2 >> zearalenone = coumestrol > genistein > daidzein > apigenin = phloretin > biochanin A = kaempferol = naringenin > formononetin = ipriflavone = quercetin = chrysin for ER alpha and E2 >> genistein = coumestrol > zearalenone > daidzein > biochanin A = apigenin = kaempferol = naringenin > phloretin = quercetin = ipriflavone = formononetin = chrysin for ER beta. Antiestrogenic activity of the phytoestrogens could not be detected, except for zearalenone which is a full agonist for ER alpha and a mixed agonist-antagonist for ER beta. In summary, while the estrogenic potency of industrial-derived estrogenic chemicals is very limited, the estrogenic potency of phytoestrogens is significant, especially for ER beta, and they may trigger many of the biological responses that are evoked by the physiological estrogens.

Unscheduled DNA synthesis in plant populations exposed to chronic irradiation.

The intensity of gamma-ray-induced unscheduled DNA synthesis (UDS) was measured in a number of natural populations of Vicia cracca which have been growing for some decades under conditions of chronic alpha- or beta-irradiation. It was shown that the level of UDS increased in more radioresistant beta-irradiated populations as compared to control populations and this increase was dose rate-dependent. In alpha-irradiated populations, the intensity of UDS was decreased, but only at the highest dose of gamma-radiation (500 Gy) and was not changed at the lower doses. The sensitivity of UDS to inhibitors of DNA and protein synthesis was studied in control and radioresistant populations. In control plants UDS was resistant to cycloheximide (Cyc) and aphidicolin (Aph), but totally inhibited by dideoxythymidine (ddT). In radioresistant population UDS was inhibited by both Aph and ddT as well as by Cyc. I assume that in controls, UDS is mediated by beta-like DNA polymerase; however, in a radioresistant population, both DNA polymerases alpha and beta take part in this process. In the radioresistant population UDS is partially inducible.

Mitochondrial bioenergetics as affected by DDT.

The organochloride insecticide DDT (2,2-bis(p-chlorophenyl)-1,1-trichloroethane) depresses the phosphorylation efficiency of mitochondria as inferred from the decrease of respiratory control ratio (RCR) and P/O ratio, perturbations of transmembrane potential (delta psi) fluctuations associated with mitochondrial energization and phosphorylative cycle induced by ADP. DDT depresses the delta psi developed by energized mitochondria and prevents complete repolarization, that is delayed and resumed at a lower rate. The inhibitory action of DDT on phosphorylation efficiency may result from: (1) a direct effect on the ubiquinol-cytochrome c segment of the redox chain; (2) direct action on the ATP-synthetase complex; (3) partial inhibition of the phosphate transporter. DDT preferentially interacts with phosphorylation process in relation to respiration. High concentrations of DDT induce destruction of the structural integrity of mitochondria.

1-(o-chlorophenyl)-1-(p-chlorophenyl)2,2,2-trichloroethane: a probe for studying estrogen and progestin receptor mediation of female sexual behavior and neuroendocrine responses.

A number of chlorinated insecticides have been shown to interact with estrogen receptors and to mimic estrogen action in peripheral reproductive tissues (e.g. vagina and uterus). The present study was designed to assess whether the dichlorodiphenyltrichloroethane (DDT) isomer o,p'-DDT has estrogenic or antiestrogenic activity in neural estrogen target tissues. Single sc injections of up to 500 mg/kg o,p'-DDT had no effect on female sexual behavior (lordosis). However, more prolonged treatments induced high levels of lordosis behavior, inhibited compensatory ovarian hypertrophy, and reduced body weight gain in ovariectomized adult female rats. Since o,p'-DDT was able to mimic the action of estrogen on these three neuroendocrine responses, further experiments were performed to determine whether the compound could be used as a tool to investigate the role of hypothalamic steroid receptors in estrogen stimulation of reproductive behavior. It was found that single injections of 500 mg/kg o,p'-DDT (which did not induce sexual receptivity) interacted with cytosol estrogen receptors in both the hypothalamus and pituitary. However, depletion of cytosol receptors by o,p'-DDT was very slow and incomplete; maximal depletion was only 51% and did not occur until 8 h postinjection. It was also observed that neither behaviorally effective nor ineffective injections of o,p'-DDT were able to induce progestin receptor synthesis in the hypothalamus or pituitary. Thus, it appears that the inability of the compound to promote lordosis behavior after a single injection probably results from inadequate receptor interaction in hypothalamic cell nuclei, but that failure to induce neural progestin receptor synthesis is not the critical factor. These data also suggest that the study of o,p'-DDT action in the brain may provide useful information regarding mechanisms of steroid mediation of neuroendocrine responses.

Role of phospholipids in the inhibitory action of DDT and permethrin on the nerve ATPase of lobster, Homarus americanus.

Efforts were made to understand the nature of the site of 1,1-bis-(p-chlorophenyl)-2,2,2-trichloroethane (DDT) inhibition of nerve ATPase. THe phospholipid content of nerve preparations from the walking leg of the lobster was reduced by treating them with phospholipase A, or with a chloroform-methanol mixture at -75 degrees. By these treatments the enzymes lost approximately 70 or 95% of their phospholipids and 50-80% of their Na,K- and Ca-ATPase activities. The lost ATPase activities could be partially restored by the addition of phospholipids, either the ones extracted from the lobster nerves or those from commercial sources. ATPase inhibition by DDT and permethrin was found to be highest in preparations where the phospholipids were removed by the above treatments, next highest with the untreated original enzyme, and least with the reconstituted ATPase regardless of the source of phospholipids used for reconstitution. This tendency was more pronounced in the case of Ca-ATPase. The effects of DDT and permethrin on inhibition of reconstituted Ca-ATPase were higher when the insecticide was first added to the protein portion and the enzyme was then reconstituted with the phospholipids, than when the same amount of insecticide was first added to the phospholipids which were then used for reconstitution.

The effect of a single dose of DDT on thyroid function in male rats.

A single dose of 100 mg/kg of commercially available 1, 1, 1,-trichloro-2, 2, bis (p-chlorophenyl), ethane, (DDT), dissolved in corn oil, was administrated to adult male Sprague-Dawley rats by stomach tube and two indices of thyroid function were examined. The iodide transport function of the thyroid gland determined by the thyroid: serum radioiodide concentration ration, (T:S) was depressed, and complete inhibition in thyroidal radioiodine release occurred 8 to 10 hr following administration of DDT and continued for 24 to 30 hr. Subsequently the block in thyroidal 131I release was followed by a return to a rate not significantly different from that of control rats. Whether the acute effects of a single large dose of DDT on thyroid function are a consequence of a local action on the thyroid gland or on pituitary thyrotropin release, or of some action elsewhere is not clear.