Abstract  The urban water system is believed to be an important sink for the nonpoint-source pollutants nonylphenols and phthalates. The presence of nonylphenols (NPs), nonylphenol ethoxylates (NPEOs), and eight phthalates was analyzed in urban stormwater and sediment from three catchment areas in Sweden. Emission loads for these substances were then calculated for a specific urban catchment area. In addition, substance distribution in road runoff passing through a sedimentation facility was modeled using a modified QWASI-model for chemical fate. High concentrations of DEHP, DIDP and DINP (

Abstract  Phthalates are synthetic compounds widely used as plasticisers, solvents and additives in many consumer products. Several animal studies have shown that some phthalates possess endocrine disrupting effects. Some of the effects of phthalates seen in rats are due to testosterone lowering effects on the foetal testis and they are similar to those seen in humans with testicular dysgenesis syndrome. Therefore, exposure of the human foetus and infants to phthalates via maternal exposure is a matter of concern. The metabolic pathways of phthalate metabolites excreted in human urine are partly known for some phthalates, but our knowledge about metabolic distribution in the body and other biological fluids, including breast milk, is limited. Compared to urine, human breast milk contains relatively more of the hydrophobic phthalates, such as di-n-butyl phthalate and the longer-branched, di(2-ethylhexyl) phthalate (DEHP) and di-iso-nonyl phthalate (DiNP); and their monoester metabolites. Urine, however, contains relatively more of the secondary metabolites of DEHP and DiNP, as well as the monoester phthalates of the more short-branched phthalates. This differential distribution is of special concern as, in particular, the hydrophobic phthalates and their metabolites are shown to have adverse effects following in utero and lactational exposures in animal studies.

Abstract  Monitoring of environmental chemicals in Japan has revealed that several endocrine active chemicals are in river water, sediments, and wildlife as well as in the human umbilical cord. In 2001, risk assessments of tributyltin and nonylphenol have been conducted by the Ministry of the Environment, Japan. Risk assessments of di(2-ethylhexyl)phthalate and di-isononyl phthalate have also been performed by the Ministry of Health, Labour and Welfare using a toxicological point of view in 2001. In this review, an overview of recent progress in endocrine disruptor research in Japan will be provided.

Abstract  Many different types of phthalate ester are used as plasticizers and are thus found in the air. There have been several studies that suggest an association between allergies and phthalate esters. We previously found that di-butyl phthalate (DBP) has an adjuvant effect in a mouse contact hypersensitivity model, in which fluorescein isothiocyanate (FITC) is involved as an immunogenic hapten. OBJECTIVE: We examined whether other phthalate esters enhance the process of sensitization to FITC by facilitating the trafficking of FITC-presenting dendritic cells or macrophages from skin sites to draining lymph nodes. METHODS: Mice were epicutaneously sensitized with FITC dissolved in acetone containing a phthalate ester. Sensitization was evaluated as ear swelling after a challenge with FITC. Draining lymph node cells obtained 24 h after skin sensitization were examined for FITC fluorescence by means of flow cytometry. FITC-positive cells were characterized with anti-CD11c and anti-CD11b by three-colour flow cytometry. RESULTS: When mice were sensitized with FITC in acetone containing DBP or di-n-propyl phthalate (DPP), strong enhancement of the ear-swelling response was observed. Di-methyl phthalate (DMP) and di-ethyl phthalate (DEP) were less effective but produced some enhancement. Consistent enhancement was not observed with di-(2-ethylhexyl) phthalate or di-isononyl phthalate. Upon sensitization in the presence of DBP or DPP, the number of FITC-positive dendritic cells (total CD11c+ as well as CD11c+/CD11b+) was increased in draining lymph nodes. As to the other four phthalate esters, there was no significant increase in the FITC-positive cell number in the draining lymph nodes. CONCLUSION: During the process of sensitization to FITC, DBP, and DPP exert strong adjuvant effects that are associated with enhancement of trafficking of antigen-presenting dendritic cells from the skin to draining lymph nodes.

Abstract  Phthalates are suspected of endocrine disrupting effects. We aimed to develop an analytical method for simultaneous determination of several phthalate metabolites in human urine, serum, and seminal plasma and to study correlations between levels of metabolites in these matrices. Thirteen metabolites were determined in samples from 60 young Danish men. Metabolites of common di-ester phthalates were detected in most urine samples. Summed di-(2-ethylhexyl) phthalate (DEHP) metabolites were excreted in urine in the highest amount (median = 91.1 ng/mL), followed by monoethyl phthalate (MEP), mono-iso-butyl phthalate (MiBP), mono-n-butyl phthalate (MnBP), mono-benzyl phthalate (MBzP), and finally summed di-isononyl phthalate (DiNP) metabolites. All these metabolite levels correlated significantly, indicating that when a participant was highly exposed to one phthalate he was also highly exposed to other phthalates. Several metabolites were also detectable in serum and in seminal plasma, although in much lower levels. Significant correlations between MEP and MiBP levels in serum and urine were observed, showing that serum levels could be used as biomarkers of human exposure. For DEHP and DiNP metabolites, correlations between urine and serum levels were only observed for mono(2-ethyl-5-carboxypentyl) phthalate (MECPP) and mono-(4-methyl-7-carboxyheptyl) phthalate (MCiOP), indicating that these secondary carboxylated metabolites were better serum markers than primary metabolites [mono(2-ethylhexyl) phthalate (MEHP) and mono-iso-nonyl phthalate (MiNP)]. In seminal plasma, only MEP levels correlated significantly to levels in urine and in serum.

Abstract  Phthalate exposure was found to be associated with endocrine disruption, respiratory effects, reproductive and developmental toxicity. The intensive use of plastics may be increasing the exposure to phthalates in Taiwanese population, particularly for young children. We studied phthalate metabolites in pregnant women and their newborns in a prospective cohort from a medical center in Central Taiwan. One hundred maternal urine samples and 30 paired cord blood and milk samples were randomly selected from all of participants (430 pregnant women). Eleven phthalate metabolites (MEHP, 5OH-MEHP, 2cx-MEHP, 5cx-MEPP, 5oxo-MEHP, MiBP, MnBP, MBzP, OH-MiNP, oxo-MiNP, and cx-MiNP) representing the exposure to five commonly used phthalates (DEHP, di-isobutyl phthalate (DiBP), DnBP, BBP, DiNP) were measured in urine of pregnant women, cord serum and breast milk after delivery, and in urine of their children. Exposure was estimated with excretion factors and correlation among metabolites of the same parent compound. Thirty and 59 urinary samples from 2 and 5 years-old children were randomly selected from 185 children successfully followed. Total urinary phthalate metabolite concentration (geometric mean, μg L⁻¹) was found to be higher in 2-years-olds (398.6) and 5-years-olds (333.7) than pregnant women (205.2). Metabolites in urine are mainly from DEHP. The proportion of DiNP metabolites was higher in children urine (4.39 and 8.31%, ages 2 and 5) than in adults (0.83%) (p<0.01). Compared to urinary levels, phthalate metabolite levels are low in cord blood (37.45) and milk (14.90). DEHP metabolite levels in women's urine and their corresponding cord blood are significantly correlated. Compared to other populations in the world, DEHP derived metabolites in maternal urine were higher, while phthalate metabolite levels in milk and cord blood were similar. The level of phthalate metabolites in milk and cord blood were comparable to those found in other populations. Further studies of health effects related to DEHP and DiNP exposure are necessary for the children.

Abstract  Amyloid beta peptide (Abeta)-induced oxidative stress may be linked to neurodegenerative disease. Ethanol extracts of Rosa laevigata protected PC12 cells from hydrogen peroxide-induced oxidative stress. (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) reduction assays revealed a significant increase in cell viability when oxidatively stressed PC12 cells were treated with R. laevigata extract. The effect of R. laevigata on oxidative stress-induced cell death was further investigated by lactate dehydrogenase release assays and trypan blue exclusion assays. Administration of 1,2-benzenedicarboxylic acid dinonyl ester from R. laevigata extract to mice infused with Abeta significantly reversed learning and memory impairment in behavioural tests. After behavioural testing, the mice were sacrificed and brains were collected for the examination of lipid peroxidation, catalase activity and acetylcholinesterase (AchE) activity. These results suggest that 1,2-benzenedicarboxylic acid dinonyl ester from R. laevigata extract may be able to reduce Abeta-induced neurotoxicity, possibly by reducing oxidative stress. Therefore, R. laevigata extract may be useful for the prevention of oxidative stress-induced neurodegenerative disorders.

Abstract  dinP is an Escherichia coli gene recently identified at 5.5 min of the genetic map, whose product shows a similarity in amino acid sequence to the E. coli UmuC protein involved in DNA damage-induced mutagenesis. In this paper we show that the gene is identical to dinB, an SOS gene previously localized near the lac locus at 8 min, the function of which was shown to be required for mutagenesis of nonirradiated lambda phage infecting UV-preirradiated bacterial cells (termed lambdaUTM for lambda untargeted mutagenesis). A newly constructed dinP null mutant exhibited the same defect for lambdaUTM as observed previously with a dinB::Mu mutant, and the defect was complemented by plasmids carrying dinP as the only intact bacterial gene. Furthermore, merely increasing the dinP gene expression, without UV irradiation or any other DNA-damaging treatment, resulted in a strong enhancement of mutagenesis in F'lac plasmids; at most, 800-fold increase in the G6-to-G5 change. The enhanced mutagenesis did not depend on recA, uvrA, or umuDC. Thus, our results establish that E. coli has at least two distinct pathways for SOS-induced mutagenesis: one dependent on umuDC and the other on dinB/P.

Abstract  Di-iso-nonylphthalate (DINP), a complex mixture of predominantly nine-carbon branched chain dialkyl phthalate isomers, has replaced di-(2-ethylhexyl)phthalate (DEHP) as the major plasticiser for polyvinylchloride (PVC) polymers. Similar to DEHP, DINP is a developmental and reproductive toxicant in rodents. This study for the first time describes human metabolism and elimination of DINP in a male volunteer after we applied a single oral DINP dose of 1.27 mg/kg body-weight. To avoid interference by omnipresent background exposure we used deuterium-labelled DINP. We investigated the urinary excretion of the simple monoester mono-iso-nonylphthalate (MINP) and oxidised isomers with hydroxy (OH-MINP), oxo (oxo-MINP) and carboxy (carboxy-MINP) functional groups. We used isomeric MINP and three specific oxidised isomer standards for quantification: mono-(4-methyl-7-hydroxy-octyl)phthalate (7OH-MMeOP), mono-(4-methyl-7-oxo-octyl)phthalate (7oxo-MMeOP) and mono-(4-methyl-7-carboxyheptyl)phthalate (7carboxy-MMeHP). These specific DINP metabolites are currently the only synthetic DINP metabolite standards available. Within 48 h we recovered 43.6% of the applied dose in urine as the above DINP metabolites, 20.2% as OH-MINP, 10.7% as carboxy-MINP, 10.6% as oxo-MINP and only 2.2% as MINP. Other oxidised DINP metabolites not determined in this study probably increase the share of the DINP dose excreted via urine. Elimination followed a multi-phase pattern, elimination half-lives in the second phase (beginning 24h post-dose) can only roughly be estimated to be 12h for the OH- and oxo-MINP-metabolites and 18 h for carboxy-MINP metabolites. After 24h, the carboxy-MINP metabolites replaced the OH-MINP metabolites as the major urinary metabolites. All oxidised DINP metabolites are suitable parameters for biomonitoring human DINP exposure.

Abstract  Di-iso-nonylphthalate (DINP) is the major plasticizer for polyvinylchloride (PVC) polymers. Two DINP products are currently produced: DINP 1 and DINP 2. We analyzed the isononyl alcohol mixtures (INA) used for the synthesis of these two DINP plasticizer products and thus identified 4-methyloctanol-1 as one of the major constituents of the alkyl side chains of DINP 1 (8.7%) and DINP 2 (20.7%). Based on this isomer, we postulated the major DINP metabolites renally excreted by humans: mono-(4-methyl-7-hydroxy-octyl)phthalate (7OH-MMeOP), mono-(4-methyl-7-oxo-octyl)phthalate (7oxo-MMeOP) and mono-(4-methyl-7-carboxy-heptyl)phthalate (7carboxy-MMeHP). We present a fast and reliable on-line clean-up HPLC method for the simultaneous determination of these three DINP metabolites in human urine. We used ESI-tandem mass spectrometry for detection and isotope dilution for quantification (limit of quantification 0.5microg/l). Via these three oxidised DINP isomer standards, we quantified the excretion of all oxidised DINP isomers with hydroxy (OH-MINP), oxo (oxo-MINP) and carboxy (carboxy-MINP) functional groups. With this approach, we can for the first time reliably quantify the internal burden of the general population to DINP. Mean urinary metabolite concentrations in random samples from the general German population (n=25) were 14.9microg/l OH-MINP, 8.9microg/l oxo-MINP and 16.4microg/l carboxy-MINP. Metabolites strongly correlated with each other over all samples analyzed (R>0.99, p<0.0001).

Abstract  Wastewater collected from oil‐water separating tanks of ten gasoline stations for a year was fractionated into diethyl ether‐soluble neutral, acidic, and basic fractions. Mutagenicity of these fractions was measured with Salmonella typhimurium strains TA98 and TA100 in the presence or absence of S9 mix. The neutral fractions showed high mutagenicity in the absence of S9 mix. Each neutral fraction was subjected to high‐performance liquid chromatography (HPLC) and fractionated. A 1‐nitropyrene(1‐NP)‐corresponding fraction was collected and analyzed by gas chromatography‐mass spectrometry (GC‐MS) and HPLC to prove that wastewater contains 1‐NP and to quantitate 1‐NP in wastewater. GC‐MS patterns showed the following molecular and fragment ion peaks of 1‐NP: 247, 217, 201, and 189. The amount of 1‐NP in 36 samples of wastewater was 4.2–25,600 ng per liter of wastewater, and 1‐NP accounted for O.3–58.5% of the total mutagenicity of the neutral fractions. The other 19 samples of wastewater did not contain any detectable 1‐NP. The mutagenicity of wastewater may be due to water from car washing and contamination by used crankcase oil. A Soxhlet extract of crankcase oil used in a gasoline engine was fractionated into three fractions as above. Mutagenicity was measured with strains TA98, TA100, TA98NR, and TA98/1,8‐DNP6 in the absence or presence of S9 mix. The neutral fraction showed the highest mutagenicity with strain TA98 in the absence of S9 mix, and its mutagenicity was decreased in strains TA98NR and TA98/1,8DNP6. The latter result indicates that the used engine‐oil contained 1‐NP and dinitropyrenes. Actually, the amounts of 1‐NP and 1,6‐diNP in used crankcase oil were 138 and 2.0 ng per ml of oil, respectively, and these concentrations accounted for 0.45 and 2.7%, respectively, of the total mutagenicity of the neutral fraction with strain TA98 in the absence of S9 mix. Moreover, the concentrations of 1‐NP and 1,6‐diNP in used crankcase oil of a diesel engine were 349 and 31 ng per ml of oil, respectively, accounting for 0.9 and 12%, respectively, of the total mutagenicity of the neutral fraction in the same assay system.

Abstract  The National Toxicology Program (NTP) Center for the Evaluation of Risks to Human Reproduction (CERHR) conducted an evaluation of the potential for di-isononyl phthalate (DINP) to cause adverse effects on reproduction and development in humans. DINP is one of 7 phthalate chemicals evaluated by the NTP CERHR Phthalates Expert Panel. These phthalates were selected for evaluation because of high production volume, extent of human exposures, use in children's products, and/or published evidence of reproductive or developmental toxicity. DINP is a mixture of branched, C-9 phthalate isomers used to add flexibility to a wide variety of plastic products such as toys, garden hoses, flooring tiles, tarps, and pool liners. The results of this evaluation on DINP are published in a NTP-CERHR monograph which includes: 1) the NTP Brief, 2) the Expert Panel Report on the Reproductive and Developmental Toxicity of Di-isononyl Phthalate, and 3) public comments received on the Expert Panel Report. As stated in the NTP Brief, the NTP reached the following conclusions regarding the possible effects of exposure to DINP on human development and reproduction. First, although DINP could possibly affect human development if exposures are sufficiently high, there is minimal concern for DINP causing adverse effects to human reproduction or fetal development. There is no direct evidence that exposure of people to DINP adversely affects reproduction or development, but studies show that oral exposure of pregnant rats to high doses (500 and 1000 mg/kg bodyweight/day) of DINP can adversely affect fetal development. Effects on pup growth were noted in a 2-generation reproductive toxicity study in rats at doses of 143-285 mg/kg body weight/day. Human exposure information for DINP was not available but it was assumed that the general US population would be exposed to 3-30 mug/kg body weight/day, based upon the range of estimated exposures for DEHP, a more widely used phthalate. Second, based on estimates of exposure of children to DINP from mouthing toys and other objects, the NTP has minimal concern for developmental effects in children. After the expert panel meeting, a US Consumer Products Safety Commission panel estimated that the majority of children exposed to DINP had a "minimum to non-existent risk of injury" from mouthing toys. Children's exposure was estimated at 70-280 mug/kg body weight/day, a level 1000-fold lower than exposures resulting in developmental effects in rats.

Abstract  In this study, samples from a sewage treatment lagoon and those from a receiving stream were analyzed for their phthalate esters content. Knowledge of the distribution of ubiquitous phthalate esters in the sewage lagoon and the receiving stream was necessary because of the reports of their subtle toxicity to aquatic biota and humans. Liquid-liquid extraction, Clean-up experiment and High Performance Liquid Chromatography (HPLC) were the methods employed for the quantitative determination of the Phthalates. A study of uncontaminated water was done to establish blank levels. The sewage lagoon and the receiving stream were grossly polluted as several phthalate ester plasticizers: DMP, DEP, DPhP, DBP, DEHP, DOP and DINP were found present at monthly mean levels of between 24.02 mg/L and 139.25 mg/L in the sewage treatment lagoon and 10.41 mg/L and 80.53 mg/L in the receiving stream. The results showed higher levels of phthalate esters in the sewage lagoon compared to the receiving stream. The sewage lagoon was identified as a pollution point source into the receiving stream. Levels of phthalates obtained from the receiving stream are much higher than the water criteria of 3 microg/L phthalates recommended by the United States Environmental Protection Agency (USEPA) for the protection of fish and other aquatic life in water and the Suggested No-Adverse Effect Levels (SNAEL) of 7.5-38.5 microg/L for drinking water. This should give cause for great environmental concern. Peoples' health downstream is at stake and so is the 'health' of the ecosystem.

Abstract  Reproductive and developmental effects of diisononyl phthalate (DINP) and diisodecyl phthalate (DIDP) were evaluated in a Japanese medaka (Oryzias latipes) multigeneration protocol. Each phthalate was administered via fish flake diets at a concentration of 20 microg/g (1 microg/g fish/day). Two controls were included, untreated and acetone carrier. The F(0) and F(1) generation adults were reared to sexual maturation and the test was ended prior to sexual maturation of the F(2) generation. Biochemical, individual, and population parameters were evaluated: testosterone metabolism, 7-ethoxyresorufin-o-deethylase (EROD) activity, survival, development, growth, gonadal-somatic index, histopathology, sex ratio, and fecundity. Male fish showed a two-fold induction of several testosterone metabolites in the DINP-treated group compared to the untreated control but not the acetone control. In a similar manner, in female fish only the DIDP-treated group expressed greater testosterone hydroxylase activity. There were neither sex- nor treatment-related differences in the results from the EROD assay. A statistically significant transient delay in red blood cell pigmentation was observed. The male-to-female ratio was consistent across treatments and the phenotypic and histological gender classifications were in agreement. Egg production was not significantly different among treatment groups. Neither phthalate elicited an effect on reproduction or development at various levels of biological organization.

Abstract  Phthalates are plasticizers that are added to polyvinyl chloride (PVC) products to impart flexibility and durability. They are produced in high volume and generate extensive though poorly defined human exposures and unique childhood exposures. Phthalates are animal carcinogens and can cause fetal death, malformations, and reproductive toxicity in laboratory animals. Toxicity profiles and potency vary by specific phthalate. The extent of these toxicities and their applicability to humans remains incompletely characterized and controversial. Two phthalates, diethylhexyl phthalate (DEHP) and diisononyl phthalate (DINP), have received considerable attention recently because of specific concerns about pediatric exposures. Like all phthalates, DEHP and DINP are ubiquitous contaminants in food, indoor air, soils, and sediments. DEHP is used in toys and medical devices. DINP is a major plasticizer used in children's toys. Scientific panels, advocacy groups, and industry groups have analyzed the literature on DEHP and DINP and have come to different conclusions about their safety. The controversy exists because risk to humans must be extrapolated from animal data that demonstrate differences in toxicity by species, route of exposure, and age at exposure and because of persistent uncertainties in human exposure data. This report addresses sensitive endpoints of reproductive and developmental toxicity and the unique aspects of pediatric exposures to phthalates that generate concern. DEHP and DINP are used as specific examples to illustrate the controversy.

Abstract  The presence of plasticizers in PVC toys obtained in October 1998 was investigated. Diisononyl phthalate (DINP), di-2-ethylhexyl phthalate (DEHP), di-n-butyl phthalate (DBP), dinonyl phthalate (DNP), diheptyl phthalate (DHP), and di-2-ethylhexyl adipate (DEHA) were detected. The phthalates were found in all of the 68 samples. The principal phthalate found in toys was DINP, which was present in 48 of 68 samples. The DINP content ranged from 15 mg/g to 580 mg/g, and mean content was 308 mg/g. The highest content was found in a pacifier toy. DEHP was present in 20 of 68 samples and the content ranged from 2.0 mg/g to 380 mg/g. The mean content was 162 mg/g. It was found in 60% of domestic toys.

Abstract  We have previously examined the impact of perinatal exposure to ethinylestradiol (EE), methoxychlor (MXC), diisononyl phthalate (DINP), and genistein (GEN) in maternal diet on rat offspring, and found developmental and/or reproductive toxicity with 0.5 ppm EE, 1200 ppm MXC, and 20,000 ppm DINP. Although the toxicological profile with MXC was similar to the EE case, the population changes in pituitary hormone-producing cells totally differed between the two cases, changes being evident from 240 ppm with MXC. In the present study, to assess the impact of these agents on brain sexual differentiation, region-specific mRNA expression of estrogen receptors (ER) alpha and beta, the progesterone receptor (PR), gonadotrophin-releasing hormone, steroid receptor coactivators (SRC)-1 and -2, and calbindin-D in microdissected hypothalamic medial preoptic areas (MPOAs) at postnatal day 10 was first analyzed in rats exposed to 0.5 ppm-EE from gestational day 15 by real-time RT-PCR. Sexually dimorphic expression of ER alpha and PR was noted with predominance in females and males, respectively, EE up-regulating SRC-1 in males and ER beta and PR in females. Next, we similarly examined expression changes of ER alpha and beta, PR, and SRC-1 in animals exposed to MXC at 24, 240, and 1200 ppm, DINP at 4000 and 20,000 ppm, and GEN at 1000 ppm. MXC at 1200 ppm down- and up-regulated PR in males and females, respectively, and DINP at 20,000 ppm down-regulated PR in females, while GEN did not exert any clear effects. The results thus suggest that agents causing developmental and/or reproductive abnormalities in later life may affect hypothalamic PR expression during the exposure period in early life.

Abstract  Methoxychlor (MXC, 24, 240, 1200 ppm), genistein (GEN, 20, 200, 1000 ppm), diisononylphthalate (DINP, 400, 4000, 20,000 ppm), 4-nonylphenol (NP, 60, 600, 3000 ppm) or bisphenol A (BA, 60, 600, 3000 ppm) were given to maternal rats from gestational day 15 to postnatal day (PND) 10 to assess their perinatal exposure effects on offsprings. Soybean-free diet was used as a basal diet. Organ weights at PND21, onset of puberty, estrous cyclicity, gonadotrophin-immunopositive index (IPI) in pituitary at PND21 and 77, and histological changes at PND77 were assessed as well as the size of sexually dimorphic nucleus of preoptic area (SDN-POA). In terms of MXC, DINP and GEN studies, expression of GABA transporter-1 (GAT-1), an estrogen responsive gene, was analyzed in medial preoptic area (MPOA) at PND10 using microdissection and real-time RT-PCR techniques. Females exposed to 1200 ppm MXC showed accelerated onset of puberty, irregular estrous cyclicity, histological changes such as multifollicular ovaries, hyperplasia in endometrium, vaginal mucosa and anterior pituitary at PND77, and decrease in LH-IPI at PND21 and increase in FSH- and PRL-IPIs at PND77. Females of 240 ppm MXC also showed increased PRL-IPI at PND77. Males of 1200 ppm MXC showed delayed onset of puberty and decreased LH-, FSH- and PRL-IPIs at PND21. DINP at 20,000 ppm caused very slight degeneration of spermatocytes and Sertoli cells at PND77. The sizes of SDN-POA did not alter at any doses of chemicals examined. GAT-1 levels in male MPOAs decreased with DINP at 20,000 ppm, and also showed a dose-related decreasing tendency with MXC. GEN, BA and NP did not affect any endocrine parameter examined. Results suggest that maternal exposure to MXC and DINP affects reproductive system of offsprings by disrupting brain sexual differentiation.

Abstract  Diisobutyl phthalate (DIBP) is the branched isomer of DBP; DBP side chains have a four-carbon backbone (C4), whereas DIBP has its four-carbon alkyl side chains rearranged into a three-carbon backbone (C3) with a methyl branch. Di-n-butyl phthalate (DBP), and several other ortho-phthalate esters with side-chain lengths of C4-C6, are known to disrupt the androgen-dependent sexual differentiation in the male rat. This study was performed to determine whether in utero exposure to DIBP would induce permanent and dose-responsive alterations of male reproductive development. Pregnant Sprague-Dawley rats were administered olive oil (vehicle control), DIBP or DBP, by gavage on gestation Days 12-21, at doses of 125, 250, 500, 625mgDIBP/(kg day) and 500mgDBP/(kg day). DIBP caused no overt maternal toxicity, nor reduced litter size. Male offspring displayed reduced neonatal anogenital distance (Postnatal day 1, PND) at 250mgDIBP/(kg day) and higher doses, and dose-related retention of areolas/nipples (PND 12-14). Preputial separation (onset of puberty) was delayed in male offspring at 500 and 625mgDIBP/(kg day). Hypospadias, cleft prepuce, and undescended testis were observed in males (11-12 or 16-17 weeks old) exposed in utero to 500 and 625mgDIBP/(kg day). Histopathological lesions were also present in adult testes, mainly consisting in seminiferous tubule degeneration. Our results show that DIBP can cause severe and specific adverse effects on the male rat reproductive development, with a pattern similar to that of DBP. However, DIBP appeared slightly less potent than DBP in inducing malformations.

Abstract  Studies of oral ingestion of environmental phthalates are reviewed. The phthalates are compared to other known substances in terms of LD50 (median lethal dose) and acceptable daily intake (ADI) values. The literature contains studies on phthalates ranging from dimethyl to ditridecyl. Acute toxicity tests define rather high LD50 values, indicating a very low order ot toxicity as compared to many common chemical substances. The acceptable ADI values calculated for phthalates from the works cited are of the same order of magnitude as some chemicals that are approved for use as direct food additives. Three instances of human ingestion were reported. In 1 of the cases (dibutyl phthalate), some toxic symptoms were reported, but the person recovered with no after-effects; the other cases (di-2-ethylhexyl phthalate) showed no toxic effects. The various studies did reach levels of administration causing toxic effects, including fatalities of some of the test species. Most investigators concluded that the phthalates constitute a chemical family of very low order to toxicity, as measured by ingestion methods. Dimethyl phthalate, the lowest molecular weight number of the family, is mildly more toxic than all of the other phthalates, but it is not considered lethal.