Exposure to the plasticizer diisopentyl phthalate can cause Vero cell line death.

Phthalates are synthetic plasticizers present in the daily lives of humans, as part of the composition of different products, such as food packaging, water bottles, and toys. These compounds can migrate from plastic materials to the environment changing biological systems. Although diisopentyl phthalate (DiPeP) is largely used in Brazil, there is a lack of information on the possible toxic effects of this compound. This research aims to evaluate the toxicity of DiPeP in the Vero renal cells. These cells were exposed to the 1-1000 µM of DiPeP for 24 and 72 h and subsequently, the cytotoxicity, apoptosis and necrosis-inducing potential, and antioxidant system (SOD, GPx, and GST) were investigated. DiPeP neither caused cytotoxicity nor altered SOD and GPx activity, although GST has been increased at 100 or 1 µM (24 and 72 h, respectively). However, cell death by apoptosis and necrosis was observed. These results indicate that DiPeP caused cell death by a non-oxidative stress-mediated mechanism, which shows the relevance of investigate other process in further researches.

Prenatal androgenization (2D:4D) predictions of tennis match-play success in junior players: A search for physiological explanations.

AIM: This study aims to investigate the possible association between digit ratio (2D:4D) and match-play success (MPS) in junior tennis players. In addition, we consider the possible explanatory pathways of these associations in relation to psychological, strength, power, and hormonal parameters. METHODS: We performed a cross-sectional study, with a sample comprised of 64 male junior tennis players (11-18 years old). Digit ratio was calculated from direct finger measurements. In addition, we measured the ratio of wins by number of matches played in 5 years of official competition (MPS), handgrip strength (HGS), standing long jump (SLJ), training (in weekly hours), and expertise (number of years in official competition). Salivary testosterone and cortisol levels were measured before and after physical "challenge" tests. RESULTS: The 2D:4D correlated negatively with HGS and SLJ. MPS was also negatively associated with 2D:4D, but was positively correlated to HGS, expertise, training, and self-confidence (SC). Multiple linear regression showed 2D:4D and expertise were associated with MPS (43%-54%). None of the physical, or hormonal variables tested explained the links between 2D:4D and MPS. CONCLUSION: Therefore, the specific fitness components influenced by prenatal androgenization that moderate sports success remain unknown. Future studies should explore the interaction of 2D:4D, with tennis exercises as a challenge to induce hormonal change, the effect of pubertal stage, and the influence of aerobic endurance in determining MPS.

Association between Late Manifestations of Testicular Dysgenesis Syndrome and Anogenital Distance: A Systematic Review and Meta-analysis.

Background: In 2001, Skakkebæk et al. proposed that certain male reproductive disorders might be grouped into a syndrome called testicular dysgenesis syndrome (TDS), as they all appear to be associated with disruption of the embryonic and foetal programming of gonadal development. TDS may be manifested in early life by the presence of genital malformations (hypospadias and cryptorchidism) and in adult life as disorders represented by low sperm counts and testicular cancer. Changes in androgen hormones during the foetal development, in addition to resulting in TDS, can also cause permanent changes in anopenile anogenital distance (AGDap) and anoscrotal anogenital distance (AGDas). Aims: The objective of this study was to determine whether there is a relationship between late manifestations of TDS and reduced anogenital/anoscrotal distance. Materials and Methods: The present study is a systematic review and meta-analysis. The research included papers from 2001 to 2020, comprising a total of 737 articles, and 13 articles were selected. Results: Linear regression analysis was performed to evaluate the relationship between the two anogenital distance measures, which showed a significant positive association (P = 0.039). A meta-analysis was also performed and compared AGDap and AGDas between control and case groups, with cases defined as men with any late TDS manifestation. These data showed a significant reduction in AGDas in the affected population (P = 0.04), but no differences in the AGDap measure (P = 0.59). Conclusion: Our study confirmed a significant relationship between reduced AGDas and late manifestations of TDS, providing further support to the association between prenatal androgen deficiency and late-onset reproductive disorders.

Evidence of genotoxicity, neurotoxicity, and antioxidant imbalance in silver catfish Rhamdia quelen after subchronic exposure to diisopentyl phthalate.

Diisopentyl phthalate (DiPeP) is a plasticizer with significant offer and application in Brazilian industries. This is attributed to its origin, which is closely linked to the refining process of sugarcane for ethanol production in the country. In this work, we developed a model for trophic exposure to environmentally relevant doses (5, 25, and 125 ng/g of DiPeP) to identify possible target tissues and toxic effects promoted by subchronic exposure to DiPeP in a Neotropical catfish species (Rhamdia quelen). After thirty days of exposure, blood, liver, kidney, brain, and muscle were collected and studied regarding DNA damage in blood cells and biochemical analyses. The kidney was the most affected organ, as in the head kidney, genotoxicity was evidenced in all groups exposed to DiPeP. Besides, the caudal kidney showed a reduction in the superoxide dismutase and glutathione peroxidase activities as well as a reduced glutathione concentration. In the liver, exposure to 125 ng/g of DiPeP increased glutathione S-transferase activity and reduced glutathione levels. In muscle, acetylcholinesterase (AChE) was reduced. However, in the brain, an increase in AChE activity was observed after the exposure to lowest doses. In contrast, a significant reduction of brain AChE activity after exposure to the highest dose was detected. The pronounced genotoxicity observed in head kidney cells is of concern, as it may compromise different functions performed by this organ (e.g., hematopoiesis, immune and endocrine functions). In our study, DiPeP proved to be a compound of environmental concern since we have evidenced its nephrotoxic and neurotoxic potential even in low doses.

Association of occupational exposure to pesticides with overweight in farmers in Southern Brazil.

INTRODUCTION: Exposure to pesticides may be related to overweight and associated comorbidities. The aim of this work was to evaluate occupational exposure to pesticides, overweight and associated comorbidities among farmers in Southern Brazil. METHODS: This cross-sectional study included a random sample of 257 farmers, living in the municipality of Mafra and Planalto, southern Brazil. Data on pesticide use and overweight prevalence from farmers were collected using an in-person interview questionnaire, followed by blood collection and biochemical analyses. RESULTS: Pesticide exposure was positively correlated with body mass index, waist circumference, waist-to-hip ratio, triglycerides and glucose levels, presence of hypertension and metabolic syndrome. Besides that, the fact of being exposed to pesticides represents a decrease of no protein thiol groups. Furthermore, the main pesticides used by farmers have hepatic toxicity. CONCLUSION: These findings suggest that exposure to pesticides may be associated with overweight and associated comorbidities. Further studies are required to validate our findings and elucidate the specific mechanisms by which these pollutants contribute to the development of overweight.

Reproductive toxicity following in utero and lactational exposure to a human-relevant phthalate mixture in rats.

This rodent (Wistar rats) study examined reproductive effects of in utero/lactational exposure to a mixture of 6 antiandrogenic phthalates (PMix): diisobutyl phthalate, di-n-butyl phthalate, diisopentyl phthalate, butylbenzyl phthalate, di-2-ethylhexyl phthalate, and diisononyl phthalate. The PMix was defined based on exposure data from pregnant women in Brazil. Experimental groups were established by extrapolating the estimated human dose to rats (0.1 mg/kg/day), followed by up to 3 additional doses corresponding to 5, 1000, and 5000 times the starting rat dose: 0 (control), 0.1, 0.5, 100, and 500 mg/kg/day. The fetal experiment assessed gestational exposure effects on fetal gonads, whereas the postnatal experiment evaluated reproductive parameters in males and females after in utero and lactational exposure. Prenatal exposure decreased fetal testicular testosterone production at 0.5 and 500 mg/kg/day. PMix 500 also reduced mRNA expression of steroidogenesis-related genes, upregulated transcript expression of the retinoic acid-degrading enzyme Cyp26b1, and increased multinucleated gonocytes incidence in fetal testes. Postnatal assessment revealed antiandrogenic effects at the highest dose, including reduced anogenital distance, nipple retention, and decreased weight of reproductive organs. Early puberty onset (preputial separation) was observed at the lowest dose in males. In contrast, females did not show significant changes in fetal and adult endpoints. Overall, the PMix recapitulated early and late male rat phthalate syndrome phenotypes at the highest dose, but also induced some subtle changes at lower doses, which warrant confirmation and mechanistic assessments. Our data support the use of epidemiologically defined mixtures for exposure risk assessments over traditional toxicological approaches.

Evaluation of the toxicity of di-iso-pentyl-phthalate (DiPeP) using the fish Danio rerio as an experimental model.

The presence of phthalates constitutes a risk to the health of aquatic environments and organisms. This work aimed to evaluate the toxic effects of di-iso-pentyl-phthalate (DiPeP) at environmentally relevant concentrations of 5, 25, and 125 µg/L in Danio rerio after subchronic exposure for 14 days. DiPeP altered the antioxidant system in the liver (125 µg/L), intestine (25 µg/L), brain, and gills in all concentrations tested. In animals exposed to 125 µg/L, DNA damage was identified in the gills. In addition, loss of cell boundary of hepatocytes, vascular congestion, necrosis in the liver, and presence of immune cells in the intestinal lumen were observed. Erythrocytic nuclear alterations in the blood occurred in animals exposed to 25 µg/L. DiPeP was quantified in muscle tissue at all exposure concentrations, appearing in a concentration-dependent manner. Contaminants such as DiPeP will still be used for a long time, mainly by industries, being a challenge for industry versus environmental health.

Exposure to phthalates and female reproductive health: A literature review.

Endocrine-disrupting chemicals (EDCs) are exogenous compounds that have been known for their ability to interfere with the action of hormones and affect endocrine pathways, including the ones involved in the development and function of both male and female reproductive systems. EDCs comprise a wide class of compounds, such as pesticides, bisphenol A, phthalates and, parabens, that are present in the environment and in several daily use products. Phthalate esters, compounds commonly used as plasticizers and additives in many industrial applications, have attracted special attention because of the widespread human exposure and the potential for disruption of androgen-dependent development in males. Although phthalates are rapidly metabolized and excreted, their ubiquitous presence ensures continuous exposures throughout different life stages from conception to adult life, as documented by a number of human biomonitoring studies worldwide. Although most research efforts have been placed on the impact of phthalates on male reproductive development and functions, there is a large body of recent experimental and observational data indicating that phthalates can negatively affect female reproductive health, and in particular alter ovarian and uterine functions, potentially contributing to disorders like polycystic ovarian syndrome, endometriosis, and other common female reproductive problems. This review summarizes the most recent experimental and epidemiologic literature on the potential effects of phthalate exposures on female reproductive health and their impact on female fertility.

The Analgesic Dipyrone Affects Pregnancy Outcomes and Endocrine-Sensitive Endpoints in Female and Male Offspring Rats.

Dipyrone is an analgesic and antipyretic drug commonly used in many countries. Although generally not recommended during pregnancy, it is known that many women use dipyrone during the gestational period. In this study, we investigated the endocrine and reproductive effects of dipyrone in female and male offspring rats exposed in utero from gestational days 10-21. Pregnant rats were treated with dipyrone at 25, 75, and 225 mg/kg/day via oral gavage. Developmental landmarks-anogenital index (AGI), number of nipples, vaginal opening, first estrus, and preputial separation-were evaluated in the offspring. Reproductive parameters, including estrous cycle regularity, daily sperm production, weight and histopathology of reproductive organs, steroid hormone levels, and gene expression of selected markers of reproductive function were assessed at adulthood. At the highest dose, dipyrone induced a significant increase in postimplantation losses/fetal death and delayed parturition in dams. Offspring exposed in utero to the highest dose also exhibited significant changes in some early life markers of endocrine disruption, in particular increased AGI in females, indicating a proandrogenic effect, and increased rate of retained nipples in males, indicating an antiandrogenic response. No changes were observed in markers of puberty onset or reproductive parameters at adulthood. These results suggest that exposure to therapeutically relevant doses of dipyrone may induce mild endocrine disruptive effects that can be detected in late pregnancy and early life. Such effects may be relevant considering dipyrone use by pregnant women and the possibility of coexposures with other endocrine disruptors.

Uterotrophic and in vitro screening for (anti)estrogenic activity of dipyrone.

Dipyrone is a commonly used analgesic in many countries and there is limited data on its possible endocrine disrupting effects. We performed a screening for in vivo and in vitro anti(estrogenic) activity of dipyrone. For the in vivo uterotrophic assay, immature female rats (22-days-old) were treated daily by oral gavage for three days with different doses of dipyrone alone (50, 100, 200 mg/kg/day) and associated with three ethynylestradiol (EE) doses (1, 3 and 10 µg/kg/day), which were based on a dose-response curve experiment. The uterine weight was used as a biomarker for estrogenicity. In a parallel in vitro approach, we used a yeast-based transcriptional activation reporter gene assay (Yeast Estrogen Screening - YES) for assessment of estrogenic agonistic and antagonistic effects of dipyrone and its main metabolites 4-methylaminoantipyrine (MAA) and 4-aminoantipyrine (AA). In the uterotrophic assay, animals that received EE at 1, 3 and 10 µg/kg/day showed an increase in relative uterine weight compared with vehicle-only rats (canola oil). Dipyrone did not increase uterine weight at any dose tested (50, 100 and 200 mg/kg/day) in relation to vehicle control, indicating absence of estrogenic activity. Furthermore, co-administration of dipyrone (50 and 200 mg/kg/day) and EE (1, 3 or 10 µg/kg/day) was unable to block EE estrogenic action in comparison to the groups treated with EE alone, indicating absence of antiestrogenic activity. In the YES assay dipyrone and its metabolites did not demonstrate estrogen agonistic or antagonistic properties in the yeast cells. These results suggest that dipyrone and its metabolites do not produce (anti)estrogenic effects in vivo or in vitro.

Could Glyphosate and Glyphosate-Based Herbicides Be Associated With Increased Thyroid Diseases Worldwide?

The increased incidence of thyroid diseases raises a series of questions about what the main predisposing factors are nowadays. If dietary restriction of iodine was once a major global health concern, today, the processes of industrialization of food and high exposure to a wide variety of environmental chemicals may be affecting, directly or indirectly, thyroid function. The homeostasis of hypothalamus-pituitary-thyroid (HPT) axis is finely regulated through the negative feedback mechanism exerted by thyroid hormones. Allostatic mechanisms are triggered to adjust the physiology of HPT axis in chronic conditions. Glyphosate and glyphosate-based herbicides are pesticides with controversial endocrine disrupting activities and only few studies have approached their effects on HPT axis and thyroid function. However, glyphosate has an electrophilic and nucleophilic zwitterion chemical structure that may affect the mechanisms involved in iodide oxidation and organification, as well as the oxidative phosphorylation in the ATP synthesis. Thus, in this review, we aimed to: (1) discuss the critical points in the regulation of HPT axis and thyroid hormones levels balance, which may be susceptible to the toxic action of glyphosate and glyphosate-based herbicides, correlating the molecular mechanisms involved in glyphosate toxicity described in the literature that may, directly or indirectly, be associated to the higher incidence of thyroid diseases; and (2) present the literature regarding glyphosate toxicity in HPT axis.

Controversies on Endocrine and Reproductive Effects of Glyphosate and Glyphosate-Based Herbicides: A Mini-Review.

Glyphosate-based herbicides (GBHs) are among the most used pesticides worldwide, presenting high potential for human exposure. Recently, a debate was raised on glyphosate risks to human health due to conflicting views over its potential carcinogenic and endocrine disruptive properties. Results from regulatory guideline studies, reports from Regulatory Agencies, and some literature studies point to a lack of endocrine disrupting properties of the active ingredient glyphosate. On the other hand, many in vivo and in vitro studies, using different experimental model systems, have demonstrated that GBHs can disrupt certain hormonal signaling pathways with impacts on the hypothalamic-pituitary-gonadal axis and other organ systems. Importantly, several studies showed that technical-grade glyphosate is less toxic than formulated GBHs, indicating that the mixture of the active ingredient and formulants can have cumulative effects on endocrine and reproductive endpoints, which requires special attention from Regulatory Agencies. In this mini-review, we discuss the controversies related to endocrine-disrupting properties of technical-grade glyphosate and GBHs emphasizing the reproductive system and its implications for human health.

The endocrine disrupting effects of sodium arsenite in the rat testis is not mediated through macrophage activation.

Arsenic (As) is an endocrine disrupting chemical that can disturb the male reproductive system. In a previous study, it was suggested that testicular macrophages could display a role in endocrine disruption induced by As exposure. This work aimed to evaluate the effects of chronic As exposure in the testis function of Wistar rats and examine the participation of macrophage activation and inflammatory response in these processes. We examined gene expression of steroidogenic machinery and immunological markers by RT-QPCR, plasma testosterone concentrations, sperm count and morphology, and histomorphometrical parameters after 60-days exposure to 1 or 5 mg.kg-1.day-1 of sodium arsenite, combined or not with 50 µg.kg-1 of LPS administered one day before euthanasia. We have demonstrated that As exposure reduced the weight of androgen-dependent organs and induced changes in spermatogenesis, in particular at the highest dose. LPS and As co-exposure promoted a decrease in testosterone synthesis, but did not increase the overexpression of markers of macrophage activation seen in LPS-only rats. Our results suggest that As does not alter the testicular macrophage function, but under immunological challenges LPS and As can display a complex interaction, which could lead to endocrine disruption.

Prenatal diclofenac exposure delays pubertal development and induces behavioral changes in rats.

Diclofenac is a non-steroidal anti-inflammatory drug widely used by the general population and, although generally contraindicated during pregnancy, it is also used by some pregnant women. This study investigated endocrine, reproductive and behavioral effects of diclofenac in male and female offspring rats exposed in utero from gestational days 10-20. Pregnant rats were treated with diclofenac at doses of 0.2, 1 and 5 mg/kg/day via oral gavage. Anogenital distance (AGD), number of nipples, and developmental landmarks of puberty onset - vaginal opening (VO), first estrus (FE) and preputial separation (PPS) - were evaluated in the offspring. At adulthood, behavioral and reproductive parameters were assessed. Male and female rats were tested in the elevated plus maze test to assess locomotor activity and anxiety-like behaviors, while male rats were also evaluated in the partner preference test. No significant effects were observed on AGD and number of nipples in both males and females. Diclofenac treatment induced an overall delay in developmental landmarks of puberty onset in male and female offspring, which reached statistical significance for PPS at the lowest diclofenac dose. Prenatal exposure to all tested doses abolished the preference of male rats for an estrous female, suggesting an impairment of brain masculinization. No changes were observed on male or female reproductive parameters at adulthood. Overall, our results indicate that prenatal exposure to therapeutically relevant doses of diclofenac may have an impact in the pubertal development of rats and negatively affect male partner preference behavior.

Prepubertal acrylamide exposure causes dose-response decreases in spermatic production and functionality with modulation of genes involved in the spermatogenesis in rats.

The increase in human infertility prevalence due to male reproductive disorders has been associated with extensive endocrine-disrupting chemical (EDC) exposure. Acrylamide (AA) is a compound formed spontaneously during heat processing of some foods that are mainly consumed by children and adolescents. In this study, we evaluated the prepubertal AA exposure effects on male adult reproductive physiology using a prepubertal experimental model to analyze the pubertal development, spermatogenesis hormones levels and genes expression involved in male reproductive function. This study is the first one to use the validated protocol to correlate the AA exposure with puberty development, as well as the AA-induced endocrine disrupting effects on reproductive axis. AA did not affect the age at puberty, the reproductive organ's weight and serum hormonal levels. AA reduces spermatogenesis, induces morphological and functional defects on sperm and alters transcript expression of sexual hormone receptors (Ar and Esr2), the transcript expression of Tnf, Egr2, Rhcg and Lrrc34. These findings suggest that excessive AA consumption may impair their reproductive capacity at adulthood, despite no changes in hormonal profile being observed.

Effects of diisopentyl phthalate exposure during gestation and lactation on hormone-dependent behaviours and hormone receptor expression in rats.

Phthalates are found in different plastic materials, such as packaging, toys and medical devices. Some of these compounds are endocrine disruptors, comprising substances that are able to induce multiple hormonal disturbances and downstream developmental effects, including the disruption of androgen-dependent differentiation of the male reproductive tract and changes in pathways that regulate hormone-dependent behaviours. In a previous study, metabolites of diisopentyl phthalate (DiPeP), a potent anti-androgenic phthalate, were found in the urine of Brazilian pregnant women. Therefore, the present study aimed to evaluate the effects of DiPeP exposure during critical developmental periods on behaviours controlled by sex hormones in rats. Pregnant Wistar rats were treated with DiPeP (1, 10 or 100 mg kg day-1 ) or canola oil by oral gavage between gestational day 10 and post-natal day (PND) 21. Male offspring were tested in a behavioural battery, including the elevated plus maze task, play behaviour, partner preference and sexual behaviour. After the behavioural tests, the hypothalamus and pituitary of these animals were removed on PND 60-65 and PND 145-160 to quantify gene expression for aromatase, androgen receptor (Ar) and oestrogen receptors α (Esr1) and ß (Esr2). Male rats exposed to 1 and 10 mg kg day-1 DiPeP displayed no preference for the female stimulus rat in the partner preference test and 1 mg kg day-1 DiPeP rats also showed a significant increase in mount and penetration latencies when mated with receptive females. A decrease in pituitary Esr1 expression was observed in all DiPeP treated groups regardless of age. A reduction in hypothalamic Esr1 expression in rats exposed to 10 mg kg day-1 DiPeP was also observed. No significant changes were found with respect to Ar, Esr2 and aromatase expression in the hypothalamus. These results suggest that DiPeP exposure during critical windows of development in rats may induce changes in behaviours related to mating and the sexual motivation of males.

In Utero and Lactational Exposure to Diisopentyl Phthalate Induces Fetal Toxicity and Antiandrogenic Effects in Rats.

A previous study has demonstrated exposure of Brazilian pregnant women to diisopentyl phthalate (DiPeP), which reduces fetal rat testosterone production in a dose-responsive manner. In this study, we examined gene expression of steroidogenic proteins in rat fetal testes and investigated the effects of in utero and lactational DiPeP exposure on male rat reproductive development and function. For the prenatal experiment, we orally exposed pregnant Wistar rats to DiPeP or di-n-butyl phthalate (reference phthalate) at 0, 125, 250, and 500 mg/kg/day from gestation day 14-18 and the fetal testis was evaluated for transcript expression of Star, Cyp11a1, Cyp17a1, Cyp19a1, Insl3, Ar, Esr1, Esr2, and Gper1 by real-time quantitative PCR (RT-qPCR). Diisopentyl phthalate lowered mRNA levels of key steroidogenic proteins, lending support to the previously reported reductions in fetal testosterone production. Diisopentyl phthalate also lowered fetal testis transcript levels of Insl3 and changed gene expression of some steroid hormones receptors. For the postnatal experiment, pregnant rats were exposed orally to vehicle (canola oil) and 4 DiPeP doses (1, 10, 100, and 300 mg/kg/day) between gestation day 10 and postnatal day 21. Diisopentyl phthalate induced a range of reproductive and antiandrogenic effects that are typical of the rat phthalate syndrome, including reduced anogenital distance at the highest dose, reduced weight of seminal vesicles at 10 mg/kg/day and above, and testicular morphological and functional changes. Signs of fetal toxicity were observed at the highest dose. Together, our results indicate that DiPeP, a compound relevant to the human exposure scenario, is one of the most active antiandrogenic phthalates.

Effects of in utero and lactational exposure to phthalates on reproductive development and glycemic homeostasis in rats.

Prenatal exposure to phthalates is associated with reproductive and metabolic systems alterations. We investigated the effects of in utero and lactational exposure to Di-(2-ethyl-hexyl) phthalate (DEHP) and Di-n-butyl phthalate (DBP) on the reproductive system and glycemic homeostasis in male and female offspring of rats. Pregnant rats were exposed to equimolar doses (0.018, 0.18 and 1.8 mmol/kg/day) of DEHP or DBP corresponding to 7, 70, and 700 mg/kg/day for DEHP and 5, 50, and 500 mg/kg/day for DBP, respectively, by oral gavage from gestation day 13 to postnatal day 21, and using canola oil as vehicle control. Male and female offspring were examined for body weight development, external markers of prenatal androgenization and puberty onset, plasma concentrations of glucose and insulin, insulin tolerance (ITT), glucose-stimulated insulin secretion (GSIS), and the expression of peroxisome proliferator-activated receptor gamma (PPARγ) and pancreatic and duodenal homeobox 1 protein (PDX-1). Male and female rats exposed to the highest doses of DEHP and DBP exhibited increased fasting glucose levels. In rats exposed to DEHP 700 mg/kg/day we also observed a reduced glucose decay rate (Kitt) following insulin administration and decreased insulin secretion in the GSIS assay. Male offspring exposed to DEHP 700 mg/kg/day had reduced anogenital distance (AGD) on PDN 4 and delayed preputial separation at puberty, while female offspring exposed to DEHP 70 and 700 mg/kg/day and to the highest DBP dose had delayed vaginal opening. Our results suggest that maternal treatment with DEHP and DBP can induce a wide range of metabolic and reproductive alterations in offspring rats, with more pronounced effects following DEHP exposure.

Unexpected, ubiquitous exposure of pregnant Brazilian women to diisopentyl phthalate, one of the most potent antiandrogenic phthalates.

BACKGROUND: Human exposure to phthalates and other non-persistent chemicals in developing countries is largely unknown. A preliminary analysis of urinary samples from pregnant Brazilian women revealed the presence of metabolites of Diisopentyl phthalate (DiPeP). OBJECTIVES: Reliably quantify DiPeP metabolites in human urine and investigate the potential antiandrogenic activity of this phthalate in rats. METHODS: We initiated a pilot pregnancy cohort in Curitiba, Brazil, to examine phthalate exposure in urine samples collected in early pregnancy (n = 50) or pooled samples from early, mid and late pregnancy (n = 44). Our well established phthalate method was modified to include the primary DiPeP metabolite, monoisopentyl phthalate (MiPeP), and two additional secondary oxidized metabolites, 3OH-MiPeP and 4OH-MiPeP. In a parallel approach, we orally exposed pregnant rats to DiPeP or Di-n-butyl phthalate (DnBP; reference phthalate) at 0, 125, 250, and 500 mg/kg/day from gestation day 14 to 18 and measured ex vivo fetal testis testosterone production. RESULTS: We were able to detect and quantify specific DiPeP metabolites in nearly all (98%) of the early pregnancy urine samples and in all gestational pool samples with a median concentration for MiPeP of 3.65 and 3.15 µg/L, respectively, and for the two oxidized metabolites between 1.00 and 1.70 µg/L. All three urinary DiPeP metabolites were strongly correlated (r = 0.89 to 0.99). In the rat model, the effective dose (mg/kg/day) inhibiting fetal testosterone production by 50% (ED50 [95% confidence interval]) was 93.6 [62.9-139.3] for DiPeP which was significantly lower than for DnBP (220.3 [172.9-280.7]), highlighting the strong antiandrogenic potency of DiPeP within the spectrum of the phthalates. CONCLUSIONS: We unveiled and confirmed the exposure of pregnant Brazilian women to DiPeP via specific urinary metabolites. This unexpected and ubiquitous DiPeP exposure indicates to unique DiPeP exposure sources in Brazil. These exposures spark considerable concern because DiPeP is one of the most potent antiandrogenic phthalates.