Tomato and Olive Bioactive Compounds: A Natural Shield against the Cellular Effects Induced by ß-Hexachlorocyclohexane-Activated Signaling Pathways.

The ß-isomer of hexachlorocyclohexane (ß-HCH) is a globally widespread pollutant that embodies all the physicochemical characteristics of organochlorine pesticides, constituting an environmental risk factor for a wide range of noncommunicable diseases. Previous in vitro studies from our group disclosed the carcinogenic potential of ß-HCH, which contributes to neoplastic transformation by means of multifaceted intracellular mechanisms. Considering the positive evidence regarding the protective role of natural bioactive compounds against pollution-induced toxicity, micronutrients from olive and tomato endowed with the capability of modulating ß-HCH cellular targets were tested. For this purpose, the solution obtained from a patented food supplement (No. EP2851080A1), referred to as Tomato and Olive Bioactive Compounds (TOBC), was administered to the androgen-sensitive prostate cancer cells LNCaP and different biochemical and cellular assays were performed to evaluate its efficiency. TOBC shows a dose-dependent significant chemoprotection by contrasting ß-HCH-induced intracellular responses such as STAT3 and AhR activation, disruption of AR signaling, antiapoptotic and proliferative activity, and increase in ROS production and DNA damage. These experimental outcomes identified TOBC as a suitable functional food to be included in a diet regimen aimed at defending cells from ß-HCH negative effects, recommending the development of tailored enriched formulations for exposed individuals.

Selection of anodic material for the combined electrochemical-biological treatment of lindane polluted soil washing effluents.

This paper focuses on the removal of lindane from soil washing effluents (SWEs) using combined electrochemical -biological processes. In particular, it has been evaluated the influence of the anodic material used in the electrolysis of the SWE on the biodegradability and toxicity of the effluents. Four anode materials were tested: Boron Doped Diamond (BDD), Carbon Felt (CF), and Mixed Metal Oxides Anodes with iridium and ruthenium (MMO-Ir and MMO-Ru). These materials were tested at different current densities and electric current charges applied. Lindane, TOC, sulphate, and chlorine species concentrations were monitored during electrochemical experiments, showing important differences in their evolution during the treatment. In spite of reaching a good removal of lindane with all the materials tested, results showed that Boron Doped Diamond working at 15 mA cm-2 achieved the best biodegradability results in the electrolyzed effluents, because the ratio BOD5/COD increased from 0.2 to 0.5, followed by Carbon Felt anode. Regarding toxicity, Carbon Felt decreased toxicity by 80%. Opposite to what it was expected, MMO anodes did not achieve biodegradability improvement and they only showed reduction in toxicity at high electrical charges.

Hexachlorocyclohexane exposure alters the microbiome of colostrum in Chinese breastfeeding mothers.

Breast milk, especially colostrum, is not just a source of nutrients and immune factors for the newborn, but also accumulates environmental persistent pollutants and its diverse microbes affect the early colonization of the newborn's gut. Little is known about associations between environmental pollutants and the microbial composition of human colostrum. We assessed the influence of hexachlorocyclohexane (HCH), a persistent organic pollutant (POP), in colostrums on the microbial composition of human colostrum samples. HCH concentrations in 89 colostrum samples collected from a population living on the easternmost island of China were measured via gas chromatography equipped with mass spectrometer (GC-MS), HCH exposure risks for infants via dietary intake of breast milk were assessed, and for 29 colostrum samples the microbiota were profiled using 16S rRNA gene pyrosequencing to assess the association with HCH exposure levels. Our study confirmed high colostrum exposure levels of total HCHs (12.19 ±â€¯13.68 µg L-1) in this Chinese population. We predominantly identified Proteobacteria (67.6%) and Firmicutes (25.1%) in colostrum and microbial diversity at the genus level differed between samples with different HCH levels; e.g., Pseudomonas which contains several HCH degrading strains was found in significantly higher abundance in γ-HCH rich samples. Also, microbes that were statistically significantly associated with HCH levels were also highly correlated with each other (false discovery rate (FDR)＜0.01) and clustered in network analysis. Microbial diversity is associated with HCH levels in human colostrum and these associations might be attributable to their HCH degrading ability. These finding provide first insights into the role that environmental persistent pollutants may play in the microbial composition of human colostrum and the colonization of the infant gut.

Organochlorine pesticides in canned tuna and sardines on the Serbian market.

The aim of this study was to determine the level of organochlorine (OC) pesticides in 57 samples of canned tuna and 31 samples of canned sardines in vegetable oil, collected from supermarkets in Serbia. OC pesticides α-HCH, ß-HCH, δ-HCH, dichlorodiphenyltrichloroethane (DDT), DDE, DDD, dielderin, endosulfane I, endosulfane II, endosulan sulfate, endrin, endrin ketone, heptachlor, heptachlor epoxide, lindane, aldrin, metoxichlor, cis-chlordane and trans-chlordane were determined using a GS-MS method. The highest concentrations (µg kg-1, arithmetic means) in canned tuna were for δ-HCH (60.6 ± 97.0) and p, p´-DDT (55.0 ± 25.1), while the corresponding values in canned sardines were for δ-HCH (90.7 ± 102.7) and endosulfane II (78.0 ± 145.9). Mean level for the sum of endosulfans was above the maximum limit in canned sardines (85.0 µg kg-1). Also, dieldrin (39.7 µg kg-1) was measured above the ML.

Prevention of hexachlorocyclohexane-induced neuronal oxidative stress by natural antioxidants.

OBJECTIVE: Decalepis hamiltonii roots are traditionally consumed as general vitalizer and used in ayurvedic medicine preparations. We have isolated/characterized potent antioxidants from the aqueous extract of the root of this plant. In this study, we examined the antioxidant potential of the aqueous extract of the roots of D. hamiltonii (DHAE) against hexachlorocyclohexane (HCH)-induced oxidative stress in four major regions of the rat brain. METHODS: The antioxidant activity of the standardized DHAE with known antioxidant constituents was tested against HCH-induced oxidative stress in the major brain regions of 60-day-old adult male Wistar rats. RESULTS: Pretreatment of rats with multiple doses of DHAE, 50 and 100 mg/kg body weight (b.w.), for 7 consecutive days significantly prevented the HCH-induced (single dose -500 mg/kg b.w.) increase in lipid peroxidation, reduction in glutathione, and altered antioxidant enzyme activities viz. superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase in major rat brain regions viz. cortex, cerebellum, midbrain, and brain stem. DHAE, per se, elevated the antioxidant status of the rat brain. DISCUSSION: DHAE shows protective action against HCH-induced oxidative stress in rat brain regions. The protective effect of DHAE could be ascribed to the isolated/characterized antioxidant compounds which could be prospective novel nutraceuticals.

Accumulation and phytoavailability of hexachlorocyclohexane isomers and cadmium in Allium sativum L. under the stress of hexachlorocyclohexane and cadmium.

The effects of Cd and HCHs with single and combined forms on Cd and HCHs phytoavailability of Allium sativum L. were investigated. The results indicated that the coexistence of Cd and HCHs presented antagonistic interactions mostly, which might be partly due to the formation of Cd-HCHs complex, compared with single stress. The bioaccumulation of Cd and HCHs in plants depended largely on their concentrations applied in pot soils, and the phytoavailability of HCH isomers was in the sequence: δ- > γ- ≥ ß- > α-HCH.

Ameliorative effect of ajwain extract on hexachlorocyclohexane-induced lipid peroxidation in rat liver.

Effect of ajwain extract on hexachlorocyclohexane-induced oxidative stress and toxicity in rats were investigated. Six groups of rats were maintained for 12 weeks as (1) Control; (2) HCH (300 mg/kg body weight) injected (3) 1% ajwain extract incorporated diet (4)1% ajwain extract incorporated diet+HCH (5) 2% ajwain extract incorporated diet and (6) 2% ajwain extract incorporated diet+HCH. Results revealed that HCH administration lead to an increase in hepatic lipid peroxidation associated with reduction in, levels of glutathione (GSH), activity of superoxide dismutase (SOD), catalase and glucose-6-phosphate dehydrogenase. Prefeeding of ajwain extract resulted in decreased hepatic levels of lipid peroxides and increased GSH, GSH-peroxidase, G-6-PDH, SOD, catalase and glutathione S-transferase (GST) activities. At the same time there was a significant reduction in hepatic levels of HCH-induced raise in lipid peroxides as a result of the prefeeding the extract. Prefeeding of ajwain extract at 1% level to rats injected with HCH reverted the significant changes in catalase, G-6-PDH, GST and -glutamyl transpeptidase. HCH-induced formation of micronuclei in femur bone marrow was also reduced significantly. It was concluded that HCH administration resulted in hepatic free radical stress, causing toxicity, which could be reduced by the dietary ajwain extract.

Protective effects of dietary ginger (Zingiber officinales Rosc.) on lindane-induced oxidative stress in rats.

The protective effect of dietary feeding of Zingiber officinales Rosc. (ginger) against lindane-induced oxidative stress was investigated in male albino rats. Oxidative stress was monitored by estimating the extent of lipid peroxidation, activities of the oxygen free radical (OFR) scavenging enzymes superoxide dismutase (SOD) and catalase (CAT) and the status of the glutathione redox cycle antioxidants. Lindane administration (30 mg/kg bw orally for 4 weeks) was associated with enhanced lipid peroxidation and compromised antioxidant defenses in rats fed a normal diet. Concomitant dietary feeding of ginger (1%w/w) significantly attenuated lindane-induced lipid peroxidation, accompanied by modulation of OFR scavenging enzymes as well as reduced glutathione (GSH) and the GSH dependent enzymes glutathione peroxidase (Gpx), glutathione reductase (GR) and glutathione-S-transferase (GST) in these rats. These findings suggest that a diet containing naturally occurring compounds is effective in exerting protective effects by modulating oxidative stress.

Attenuation of the effect of lindane on immune responses and oxidative stress by Ocimum sanctum seed oil (OSSO) in rats.

Present study was done to evaluate the effect of Ocimum sanctum seed oil (OSSO) on the immunotoxicity and oxidative activity of lindane in rats. Rats were divided into four groups (n = 8) and were treated with lindane (10 mg/kg, po) and/or OSSO (1 mg/kg, po) during the study period. Humoral immunity was assessed by measuring haemagglutination titre to sheep red blood cells (SRBC) and delayed type hypersensitivity (DTH) was assessed by measuring foot pad thickness. Lindane showed significant decrease in anti-SRBC antibody titre and also decreased percentage change in foot pad thickness in DTH response as compared to control group. OSSO per se produced significant increase in anti-SRBC antibody titre, but did not produce significant change in the foot pad thickness as compared to control group. However, it significantly antagonized the effect of lindane on the anti-SRBC antibody titre and foot pad thickness parameters. Lindane produced oxidative stress as indicated by increase in the levels of MDA and decrease in GSH levels. Treatment with OSSO per se showed antioxidant activity and also reversed the oxidative stress produced by lindane. The results suggest that OSSO can attenuate the immunotoxicity and oxidative stress produced by lindane.

PolicosanolPlus and Neuroprevin ameliorate pesticide-mediated inhibition of neurite outgrowth and neurite degeneration.

BACKGROUND: Pesticide exposure is a recognized risk factor for neurodegenerative diseases. Recently, bifenthrin, a pyrethroid pesticide, was shown to inhibit the formation of neurites and cause neurite retraction, raising concern that these newer and less toxic pesticides may also contribute to neurodegenerative diseases. PolicosanolPlus and Neuroprevin are nutraceutical supplements which promote the survival of neurites in neuronal cell cultures. Here we determine if PolicosanolPlus and Neuroprevin can ameliorate the neurodegenerative effects of bifenthrin. MATERIAL/METHODS: PC12 cells were treated with NGF, bifenthrin, PolicosanolPlus and Neuroprevin in various combinations and the formation of neurites was assessed microscopically at times ranging from 12 to 72 hours post treatment. Bifenthrin was also withheld at the time of NGF, PolicosanolPlus and Neuroprevin treatment and added after neurite formed to assess neurite retraction. RESULTS: Bifenthrin (1 x 10(-6) M) inhibits neurite outgrowth, in the absence of cell death, by more than 50% at 12 hours and by more than 80% at 72 hours. With addition of PolicosanolPlus and/or Neuroprevin at the time of cell seeding, bifenthrin does not inhibit neurite outgrowth. Addition of bifenthrin to differentiated cells results in a retraction of 90% of neurites, while those with PolicosanolPlus and Neuroprevin show no significant retraction of neurites. CONCLUSIONS: The pesticide, bifenthrin, inhibits neurite formation and causes neurite retraction. PolicosanolPlus and Neuroprevin are nutraceutical supplements which ameliorate the effects of bifenthrin on neurite outgrowth and retraction. Dietary supplementation with PolicosanolPlus and Neuroprevin may protect against developmental and long-term neurodegenerative events that result from exposure to pesticides.

The protective effect of aloe vera juice on lindane induced hepatotoxicity and genotoxicity.

The protective effect of fresh aloe vera (AV) leaves extract on lindane (LD) - induced hepatoxicity and genotoxicity was studied. Serum levels of hepatic enzyme markers: glutamate pyruvate transaminase (GPT), glutamate oxaloacetate transaminase (GOT), gamma glutamyl transferase (GGT) and alkaline phosphatase (ALP) were determined after oral administration of aloe vera leaves extract and lindane. The level of polychromatic erythrocytes was also observed. Pretreatment with aloe vera leaves extract at concentration of 1.0 ml/kg body weight significantly decreased (P<0.05) the serum levels of GPT (by 41.8%), GOT (by 36.5%), GGT (by 14.3%) and ALP (by 10.7%) induced by 100mg/kg body weight of lindane. The level of polychromatic erythrocytes observed was not statistically significant when compared to control.

Hepatoprotective and antioxidant property of Andrographis paniculata (Nees) in BHC induced liver damage in mice.

Andrographis paniculata (AP) treatment prevents BHC induced increase in the activities of enzymes y-Glutamyl transpeptidase, glutathione-S-transferase and lipid peroxidation. The activities of antioxidant enzymes like superoxide dismutase, catalase, glutathione peroxidase and the levels of glutathione were decreased following BHC effect. Administration of AP showed protective effects in the activity of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase as well the level of glutathione. The activity of lipid peroxidase was also decreased. The result indicate antioxidant and hepatoprotective action of A. paniculata.

Acute marine sediment toxicity: a potential new test with the amphipod Gammarus locusta.

Although amphipod toxicity tests have been successfully used in the United States to assess coastal sediment toxicity, few tests have been developed with European species. The authors have been working with the amphipod Gammarus locusta, a widely spread species along European coastal areas that is particularly abundant in the Portuguese Sado estuary. This amphipod fulfills the most important requirements of a test species. It can be easily reproduced in laboratory and it is tolerant to a broad range of sediment types. A series of tests demonstrated its sensitivity to copper and gamma-hexachlorocyclohexane (lindane) in the sediment (LC50 = 6.8 mg Cu/dry kg, 0.9% total volatile solids; LC50 = 60.5 micrograms HCH/dry kg, 2% total volatile solids) and to some heavily contaminated field sediments. After assessment of the species sensitivity to several noncontaminant variables, an experimental protocol was designed to conduct acute sediment toxicity tests that are briefly described. Proposed is a 10-day static toxicity test at 15 degrees C and 33-34/1000 salinity, with laboratory-produced juveniles and mortality as the endpoint. General assay performance is identical to the American Society for Testing and Materials (ASTM) standard for sediment toxicity tests with marine and estuarine amphipods. The results previously obtained revealed a strong potential for this amphipod to be used in toxicological testing. Considering the wide geographic distribution of this species and its amenability for culturing, it may be an alternative or complementary test for ecotoxicological studies in other European coastal systems where the existing tests cannot be applied or do not offer a definitive solution.

Progress in an ecotoxicological standard protocol with protozoa: results from a pilot ring test with Tetrahymena pyriformis.

The aim of this project was to develop a standardized multigeneration growth inhibition test protocol to assess the sublethal effects of industrial chemicals and pesticides on protozoans. The inclusion of a test with protozoa would be an ecologically relevant supplement to the existing basic set in aquatic toxicity testing. The different aspects evaluated in the pilot ring study include: two media, two volumes, two end points and two different methods to measure the cell density. Taking into account the aim to develop a simple, sensitive, reproducible and cost effective test, recommendations are made for a standardized protocol.

In vitro effects of gamma-hexachlorocyclohexane on in vitro biosynthesis and metabolism of steroids in goldfish Carassius auratus.

Testicular and ovarian fragments of Carassius auratus, taken during the reproductively active prespawning phase (June) of its annual reproductive cycle, were incubated with different concentrations (0 mg/ml, 0 ppm; 0.001 mg/ml, 1 ppm; 0.01 mg/ml, 10 ppm; and 0.02 mg/ml, 20 ppm) of gamma-hexachlorocyclohexane (gamma-HCH) in the presence of either exogenous precursor [3H]-17-hydroxyprogesterone ([3H]17-P) or carp hypophyseal homogenate. The free (unconjugated) and conjugated metabolites (glucuronides and sulfates) of [3H]-17-P [androstenedione (AD), androstenetrione, 17-hydroxyprogesterone, testosterone (T), 11-deoxycortisol (S), 17, 20alpha-dihydroxy-4-pregnen-3-one (17,20alphaP), 17, 20beta-dihydroxy-4-pregnen-3-one (17,20betaP), 7alpha-pregnanetetrols (7alpha-P), and other polar metabolites] were separated by thin-layer chromatography and high-performance liquid chromatography. The endogenous production of unconjugated (free) steroids T, 17,20betaP, S, and 11-ketotestosterone (11-KT) in response to gamma-HCH were measured by radioimmunoassay. Among the in vitro metabolism of [3H]-17-P, in males, free steroids of AD, T, 17,20alphaP, S, and polar-free steroids were increased with the decreased yield of 11-KT. Percentage yield of testosterone glucuronide (TG) was increased with highly significant decreased yields of polar glucuronide steroids. The sulfate steroids of 17, 20alphaP, 17,20betaP, S, and 11-KT remain unchanged. In females, the decreased percentage of yield of AD and S and elevated T were noticed. The yield of TG was increased with decreased yield of 7alpha-P glucuronides. The percentage yield of AD sulfate and sulfate steroids of 17,20alphaP, 17,20betaP, and S were noted to be increased, but the yield for S sulfate was very high. Endogenous production of T was increased in both sexes in the presence of gamma-HCH, but 11-KT in male and S in female were depressed. 17, 20betaP was stimulated at some concentrations in both sexes but levels were very low. Results indicate that gamma-HCH in vitro perturbed the steroid biosynthesis during this phase thereby affecting reproductive physiology.

Toxicology of cupric salts in honeybees. I. Hormesis effects of organic derivatives on lethality parameters.

Feeding bees with organic cupric salts provides long-term control of the parasite Varroa jacobsoni. A set of new algebraic parameters (M. Bounias C.R. Acad. Sci. 310(3), 65-70, 1990) completely describing the population lethality function has been calculated following chronic administration of cupric gluconate, aspartate, and isoleucinate, with or without dietary pollen. Mortality curves allowed the calculation of LT50 (time for 50% lethality) as well as Hill coefficients (h) of the curves and the LD50 as a function of time. The tangent at the inflexion point of the sigmoidal time/mortality curves (delta i) gave the maximum mortality acceleration as an additional parameter. No toxicity (i.e., no decrease of TL50 vs doses and no LD50 values) was found for cupric gluconate and isoleucinate with pollen, whereas increases in LT50 and decreases in delta indicated hormesis effects. Doses decreasing by half-time LT50, h, or delta were used as objective lethality indexes for comparisons of toxicity in the other cases. Routine acute toxicity at high dosage was also compared with phosalone and lindane effects 24 hr after treatment.

Lindane does not alter the estrogen receptor or the estrogen-dependent induction of progesterone receptors in sexually immature or ovariectomized adult rats.

Lindane, gamma-1,2,3,4,5,6-hexachlorocyclohexane (gamma-HCH), has been shown to disrupt reproductive function in mammals. Many of these adverse effects on female reproduction such as alterations in sexual receptivity, disrupted ovarian cyclicity, reduction in uterine weight and termination of pregnancy are thought to be due to altered ovarian hormone secretions and/or an impaired response to circulating estrogen. It has been suggested that gamma-HCH may block the response of estrogen-dependent tissues to estradiol via an interaction with the estrogen receptor. To test this hypothesis, estrogen (ER) and progesterone (PR) receptor affinity and number were evaluated in sexually immature, 17 beta-estradiol-3-benzoate (EB)-primed Long Evans female rats following exposure to vehicle or gamma-HCH (40 mg/kg) for 7 days (Study 1) and in adult, ovariectomized EB-primed Long-Evans rats following gavage with vehicle or gamma-HCH (0, 10, 20, or 40 mg/kg) for 5 days (Study 2). Chlordecone (kepone; 40 mg/kg; i.p.) was used in Study 2 as a positive control for the alteration of the estrogen-induction of PR in the pituitary. Neither gamma-HCH nor chlordecone altered serum estradiol concentrations. gamma-HCH did not change the ER number (1, 24, or 30 h after EB) or the estrogen-dependent induction of PR (24 or 48 h after EB) in the hypothalamus (HYP), pituitary, or uterus. These data indicate that the effects of gamma-HCH on the female reproductive system do not involve an alteration in the ER and that heterogeneity exists between target tissues in their response to xenobiotics.