Abstract  Di-isononyl phthalate (DINP; CAS no. 68515-48-0) is a general-purpose plasticizer for polyvinyl chloride. It produced liver and kidney effects when given to rodents at high oral doses, but there were no target organ effects in primates treated under similar conditions. To assist in understanding the basis for these species differences, the pharmacokinetic properties of DINP were evaluated in rodents following both oral and dermal administration. These studies demonstrated that the pharmacokinetic properties of DINP are similar to those of other high-molecular-weight phthalates. When orally administered to rodents, DINP is rapidly metabolized in the gastrointestinal tract to the corresponding monoester, absorbed and excreted, primarily in the urine. Shortly after administration, DINP is found primarily in liver and kidneys, but it does not persist or accumulate in any organ or tissue. It is very poorly absorbed from the skin, but once absorbed it behaves in the same way as the orally administered material. The results of these rodent studies contrast with data from studies involving humans or other primates, which indicate low absorption at low oral doses and much more limited total absorption at high doses. It appears that many, if not all, of the effects of DINP in rodent studies are associated with internal doses that would be difficult, if not impossible, to achieve in humans under any circumstances. Thus, the results of rodent studies may not be very useful in assessing the potential risks to humans from high-molecular-weight phthalates.

Abstract  Recently there have been reports of liver and kidney tumors in rodents following long-term exposure to di(isononyl) phthalate (DINP). Mechanistic studies suggested that the liver tumors were a consequence of peroxisomal proliferation, whereas the kidney tumors (found only in male rats) were associated with induction of alpha(2u)-globulin. Because both peroxisomal proliferation and alpha(2u)-globulin are considered to be non-genotoxic carcinogenic processes, it seemed appropriate to investigate the genotoxic potential of DINP. Additional studies were also conducted on di(isodecyl) phthalate (DIDP), a structurally related substance that also induces peroxisomal proliferation, although it has not been tested in a carcinogenicity bioassay. The DINP was tested in Salmonella, in vitro cytogenetics and mouse micronucleus assays, whereas DIDP was evaluated in a mouse micronucleus test. All of these tests produced negative results, i.e. neither phthalate was mutagenic in any of the test systems. These data are consistent with results of other published and unpublished genotoxicity tests and provide support for the hypothesis that the liver and kidney tumors induced by DINP were the result of non-genotoxic processes.

Abstract  Macrophage activation contributes to adverse effects produced by a number of hepatotoxic compounds. Transcriptional profiles elicited by two macrophage activators, LPS and zymosan A, were compared to those produced by 100 paradigm compounds (mostly hepatotoxicants) using cDNA microarrays. Several hepatotoxicants previously reported to activate liver macrophages produced transcriptional responses similar to LPS and zymosan, and these were used to construct a gene signature profile for macrophage activators in the liver. Measurement of cytokine mRNAs in the same liver samples by RT-PCR independently confirmed that these compounds are associated with macrophage activation. In addition to expected effects on acute phase proteins and metabolic pathways that are regulated by LPS and inflammation, a strong induction was observed for many endoplasmic reticulum-associated stress/chaperone proteins. Additionally, many genes in our macrophage activator signature profile were well-characterized PPARalpha-induced genes which were repressed by macrophage activators. A shared gene signature profile for peroxisome proliferators was determined using a training set of clofibrate, WY 14643, diethylhexylphthalate, diisononylphthalate, perfluorodecanoic acid, perfluoroheptanoic acid, and perfluorooctanoic acid. The signature profile included macrophage activator-induced genes that were repressed by peroxisome proliferators. NSAIDs comprised an interesting pharmacological class in that some compounds, notably diflunisal, co-clustered with peroxisome proliferators whereas several others co-clustered with macrophage activators, possibly due to endotoxin exposure secondary to their adverse effects on the gastrointestinal system. While much of these data confirmed findings from the literature, the transcriptional patterns detected using this toxicogenomics approach showed relationships between genes and biological pathways requiring complex analysis to be discerned.

Abstract  We have investigated the effects of several phenols (octylphenol [OP], nonylphenol [NP], tert-octylphenol [tOP]) and phthalates (dioctylphthalate [DOP], diisodecylphthalate [DiDP], diisononylphthalate [DiNP]) on steroid hormone production by porcine ovarian granulosa cells after a 72-hour incubation. These chemicals are widely used as plasticisers and are suspected to possess endocrine disrupting properties. No changes were exhibited in basal progesterone production after treatment with NP or tOP, or with the tested phthalates. However, OP tended to decrease progesterone levels, while DOP and DiDP, at the lowest concentration used (10(-8)M), increased progesterone levels in the culture media. Neither of the tested phenols affected follicle stimulating hormone (FSH)-stimulated progesterone production, except for OP and NP at 10(-4)M, which decreased progesterone levels. The phthalates, tested at higher concentrations, were able to amplify FSH-stimulated progesterone release into the culture medium. An inhibitory action on oestradiol production by porcine granulosa cells was observed after the treatment with both groups of test chemicals. The results obtained in the experiments on primary granulosa cell cultures indicate that ovarian steroidogenesis might be one of the possible sites affected by the endocrine disrupting actions of phenols and phthalates.

Abstract  With Swedish summary.. The objectives of the study were to produce reliable 'background' data of phthalate concentrations in sediments, and to assess the relative importance of phthalate input from both diffuse sources and from point-sources. Much effort was first concentrated on adopting new laboratory and sampling practices with a minimum of contamination risks. Thereafter, surface sediments (0-2 cm) from different types of environments were sampled and analysed for the following nine substances and mixtures: DMP, DEP, DEHP, DBP, BBP, DnOP, DnNP, DINP and DIDP. The sampling stations were selected in order to represent: * 'background' concentrations from different parts of Sweden (remote lakes), * a gradient of increasing anthropogenic influence, downstream of a river system and * water-bodies acting as recipients of 'points-sources' (plastic industries). One sediment core was also analysed for depth profiles of phthalates. DEHP varied between 0.01 and 0.4 (mu)g/g dw in the remote lakes. The concentrations of DBP and BBP were 0-0.9 (mu)g/g dw while the other phthalates were mostly undetectable. In the gradient with increasing anthropogenic influence, DEHP concentrations (normalised to organic content in the sediments), increased downstream. The DEHP-concentrations at the two points sources were 33 and 47 (mu)g/g dw respectively, which represent more than a ten-fold decrease compared to concentrations measured in 1983. Comparatively high concentrations of the phthalate mixture DINP was measured in the sediments at one of the points sources. 17 refs, 7 figs, 4 tabs

Abstract  Nineteen samples of food in glass jars with twist closures were collected by the national food inspectors at Danish food producers and a few importers, focusing on fatty food, such as vegetables in oil, herring in dressing or pickle, soft spreadable cheese, cream, dressings, peanut butter, sauces and infant food. The composition of the plasticizers in the gaskets was analysed by gas chromatography with flame ionization detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Epoxidized soybean oil (ESBO) and phthalates were determined in the homogenized food samples. ESBO was the principal plasticizer in five of the gaskets; in 14 it was phthalates. ESBO was found in seven of the food samples at concentrations from 6 to 100 mg kg(-1). The highest levels (91-100 mg kg(-1)) were in oily foods such as garlic, chilli or olives in oil. Phthalates, i.e. di-iso-decylphthalate (DIDP) and di-iso-nonylphthalates (DINP), were found in seven samples at 6-173 mg kg(-1). The highest concentrations (99-173 mg kg(-1)) were in products of garlic and tomatoes in oil and in fatty food products such as sauce bearnaise and peanut butter. For five of the samples the overall migration from unused lids to the official fatty food simulant olive oil was determined and compared with the legal limit of 60 mg kg(-1). The results ranged from 76 to 519 mg kg(-1) and as a consequence the products were withdrawn from the market.

Abstract  The levels of dibutyl phthalate (DBP), butylbenzyl phthalate (BBP), di-(2-ethylhexyl) phthalate (DEPHP), di-isononyl phthalate (DINP), di-(2-ethylhexyl) adipate (DEHA) and di-isononyl adipate (DINA) were determined in 50 processed foods (ham and sausage, fried dumpling and shao-mai, fish paste products, croquette and fried fish, bread, noodle, pickles, etc.). DBP, BBP, DEHP, DINP, DEHA, and DINA were contained at nd approximately 47.7, nd approximately 16.6, nd approximately 749, nd approximately 358, nd approximately 57.2 and nd approximately 20,200 ppb, respectively. High-level contamination of DINA was found in fish paste products, croquette and shao-mai, presumably because of migration from plasticized wrapping film using for food packaging. We studied the relationship between DINA migration from wrapped PVC film into fried croquette and its standing time after frying. When the croquette was wrapped immediately after frying, the migration from wrapping film into the croquette was highest (36,400 ng/g). On wrapping after standing for 5 min and 30 min, the migration level was reduced to 1/3.5 and 1/14 of the highest level, respectively.

Abstract  BIOSIS COPYRIGHT: BIOL ABS. RRM DIPLORIA-STRIGOSA ARCA-ZEBRA GILL HEPATOPANCREAS MUSCLE TOXICITY ATMOSPHERIC FALLOUT BERMUDA

Abstract  This study was intended to provide data on the composition of soft PVC toys, addressing the widest practicable range of chemical additives and including non-phthalate additives. The study also included toys from as many countries as possible, since for many, no data were available. A total of 72 toys were purchased in 17 countries. The majority (64) were PVC or had PVC sections. In almost all the soft PVC toys analysed, phthalates comprised a sizeable proportion (most frequently 10-40%) of the total weight of the toy.The predominant phthalates detected were diisononyl phthalate (DINP) and di(2-ethylhexyl) phthalate (DEHP). Other phthalates identified in high concentrations in some toys include isomeric mixes of diisooctyl phthalate (DIOP) and diisodecyl phthalate (DIDP). The estrogenic chemical nonylphenol was isolated from 13 toys, while 2 toys were found to contain the fungicide Fungitrol 11 (Folpet). 78% of PVC toys contained one or more extractable organic compounds in addition to those reported above.

Abstract  Daily oral exposure of babies to phthalate was estimated on the basis of the mouthing time of infants and the oral concentration of diisononyl phthalate (DINP) released from polyvinyl chloride (PVC) toy specimens. Total mouthing time, including the use of pacifiers, ranged widely from 11.4 min to 351.8 min with the mean of 105.3 +/- 72.1 min. The mean of the total mouthing time without pacifiers was 73.9 +/- 32.9 min. The average amount of DINP in saliva was 92.4 +/- 56.8 micrograms/10 cm2/hr, ranging from 13.2 micrograms/10 cm2/hr to 240.4 micrograms/10 cm2/hr. The exposure of phthalate in two different trials was estimated by the method of Monte Carlo simulation, one with the total mouthing time with pacifiers and the other with the total mouthing time without pacifiers. The average exposure in the former trial was 21.4 micrograms/kg/day and the 95th percentile was 65.8 micrograms/kg/day, while in the latter it was 14.8 micrograms/kg/day and the 95th percentile was 35.7 micrograms/kg/day.

Abstract  Plasticizers in duplicate diet samples obtained over 1 week were analysed in order to estimate daily intake. The phthalate esters were as follows: diethyl, dipropyl, dibutyl, dipentyl, dihexyl, butylbenzyl, dicyclohexyl, di(2-ethylhexyl), dioctyl, diisooctyl (mixture of isomers) and diisononyl (mixture). Di(2-ethylhexyl) adipate was also determined. Homogenized samples of composite meals were extracted with acetonitrile, lipids were removed by extraction into n-hexane and the acetonitrile layer was cleaned using Florisil and Bondesil PSA dual layer column. Phthalates were determined by GC/MS (SIM). Phthalate recovery from the fortified food mixture by this method was 62.5-140.8%. Quality assurance as assessed by three laboratories indicated coefficient of variance in the levels of detected phthalates in same lot samples as below 10%. Detection limits were 0.1-23 ng/g for each phthalate. One-week duplicate diet samples provided by three hospitals in three remote prefectures of Japan were analysed as individual meals. In all 63 samples, DEHP was present at the highest level among all phthalates in the range 10-4400 ng/g. The intake of plasticizers estimated from all samples was 519 microg DEHP/day, 86 microg DEHA/day, 65 microg DINP/day, and 4.7 microg BBP/day. Calculated DEHP in 2-day samples out of 21 days exceeded EU TDI for a person of 50 kg body weight (1850 microg per day). Disposable PVC gloves used during the preparation of meals were suspected as the source of the high DEHP content. One-day intake of the other phthalates and DEHA was below 7% of TDI in all cases. High concentrations of DEHP (5990 ng/g) was found in baby food used in quality assurance work. The source of contamination was the PVC-tube used during production and was effectively reduced by replacing the tube by one made of stainless steel.

Abstract  Duplicate hospital diet samples obtained over 1 week in 2001 were analysed to estimate the daily intake of plasticizers and the results were compared with those obtained in 1999. The plasticizers quantified in this study were: dibutyl phthalate, butylbenzyl phthalate (BBP), di(2-ethylhexyl) phthalate (DEHP), diisononyl phthalate (DINP), di(2-ethylhexyl) adipate (DEHA), diisononyl adipate (DINA) and O-acetyl tributyl citrate (ATBC). Dipropyl, dipentyl, dihexyl and dicyclohexyl phthalate were also analysed but not detected. The analytical procedure for this follow-up study was essentially the same as in the previous one. Detection limits were 0.1-15.6 ng g(-1) for each plasticizer. One-week duplicate diet samples provided by three hospitals in three remote prefectures of Japan were analysed as individual meals. DEHP was detected at 6-675 ng g(-1) in 62 of 63 meals, significantly lower levels compared with those detected in 1999. Levels of DEHA and DINP also decreased. The mean intake of plasticizers estimated from all samples was 160 microg DEHP day(-1), 12.5 microg DEHA day(-1), 4.7 microg DINP day(-1) and 3.4 microg BBP day(-1). Levels of DINA were relatively high in meals from one hospital: in those meals, the average daily intake was 1338 microg day(-1). Those of ATBC were also higher in meals from another hospital: the average daily intake was 1228 microg day(-1). The sources of DINA and ATBC can be cling-film or sausage packaging.

Abstract  The developmental toxicity of di-isodecyl phthalate (DIDP; CAS RN 68515-49-1) and di-isononyl phthalate (DINP; CAS RN 68515-48-0) were investigated in Sprague-Dawley rats. DIDP and DINP were administered by gavage to mated rats at doses of 0, 100, 500, and 1000 mg/kg/d on Gestation Days (GD) 6 through 15. Cesarean sections were performed on GD 21 and the fetuses removed for evaluation. Maternal weight gain and food consumption were significantly reduced at 1000 mg/kg/d during the exposure period. No treatment-related effects were noted at cesarean section, nor were there any fetal morphologic observations except for an increased frequency of seventh cervical and rudimentary lumbar ribs at the maternally toxic exposure level of 1000 mg/kg/d. Under these study conditions, mild maternal and developmental effects were observed at 1000 mg/kg/d. Both maternal and developmental NOAELs were therefore established at 500 mg/kg/d. The results indicate that neither DIDP nor DINP is teratogenic or a selective developmental toxicant.

Abstract  The potential reproductive toxicity of di-isononyl phthalate (DINP: CAS RN 68515-48-0) was assessed in one- and two-generation reproductive toxicity studies. Groups of 30 male and female CRL : CD(SD)BR rats were given DINP via dietary administration at levels of either 0.0, 0.5, 1, or 1.5% (one-generation study) or 0.0, 0.2, 0. 4, or 0.8% (two-generation study). There were no changes in any of the classic reproductive parameters, i.e. mating, male or female fertility, fecundity, gestational index, or length of gestation in either study. The overall NOAELs for these effects were the highest Dietary Level (%)s tested, approximately 500 mg/kg/day in the two-generation study and 1000 mg/kg/day in the one-generation study. There were no testicular effects in parental animals exposed as juveniles and young adults at 960 mg/kg/day in the one-generation study. In the two-generation study, there were no testicular effects in either the P(1) males, exposed as juveniles and young adults or the P(2) (F(1)) offspring exposed in utero, through lactation, and continuously to terminal sacrifice. The NOAEL was 470 mg/kg/day. Offspring survival was reduced at the 1.5% level ( approximately 1100 mg/kg/day) but unaffected at the 1% level ( approximately 760 mg/kg/day). There were decreased offspring body weights both at postnatal day (PND) 0 and during lactation; however, the PND 0 effects were only clearly related to treatment at the 1.5% level. Weights of offspring during lactation were significantly reduced but within the historical control range at Dietary Level (%)s below 1%. As there was rapid recovery at the lower levels, even though treatment continued, the toxicologic significance is unclear. Adult survival was unaffected at any level in either study, but weight gain was significantly reduced at the 1% level ( approximately 600 mg/kg/day). Liver and kidney weights were elevated at Dietary Level (%)s above approximately 110 mg/kg/day, consistent with evidence from other studies of peroxisomal proliferation at these levels. This study showed that DINP treatment does not affect fertility or male reproductive development at doses of up to approximately 1000 mg/kg/day.

Abstract  BIOSIS COPYRIGHT: BIOL ABS. This study describes organic compounds in the flue gas of a hazardous waste incinerator. Polycyclic and heterocyclic aromatics, phthalate esters, phosphate esters, halogenated benzenes, biphenyls, naphthalenes, phenols as well as nitro compounds were separated by capillary gas chromatography, characterized and quantified by mass spectroscopy.

Abstract  Phthalic acid esters (phthalates) are used as plasticizers in numerous consumer products, commodities, and building materials. Consequently, phthalates are found in human residential and occupational environments in high concentrations, both in air and in dust. Phthalates are also ubiquitous food and environmental contaminants. An increasing number of studies sampling human urine reveal the ubiquitous phthalate exposure of consumers in industrialized countries. At the same time, recent toxicological studies have demonstrated the potential of the most important phthalates to disturb the human hormonal system and human sexual development and reproduction. Additionally, phthalates are suspected to trigger asthma and dermal diseases in children. To find the important sources of phthalates in Europeans, a scenario-based approach is applied here. Scenarios representing realistic exposure situations are generated to calculate the age-specific range in daily consumer exposure to eight phthalates. The scenarios demonstrate that exposure of infant and adult consumers is caused by different sources in many cases. Infant consumers experience significantly higher daily exposure to phthalates in relation to their body weight than older consumers. The use of consumer products and different indoor sources dominate the exposure to dimethyl, diethyl, benzylbutyl, diisononyl, and diisodecyl phthalates, whereas food has a major influence on the exposure to diisobutyl, dibutyl, and di-2-ethylhexyl phthalates. The scenario-based approach chosen in the present study provides a link between the knowledge on emission sources of phthalates and the concentrations of phthalate metabolites found in human urine.

Abstract  The purpose of this study was to investigate the relationship between the levels of prenatal exposure to phthalate ester and PAHs and birth outcomes among 149 Japanese pregnant women. Urinary concentrations of 9 phthalate ester metabolites, mono methyl phthalate (MMP), mono ethyl phthalate (MEP), mono-n-butyl phthalate (MnBP), mono benzyl phthalate (MBzP), mono-2-ethylhexyl phthalate (MEHP), mono-2-ethyl-5-hydroxyhexyl phthalate (MEHHP), mono-2-ethyl-5-oxohexyl phthalate (MEOHP), mono-iso-nonyl phthalate (MINP) and mono-n-octyl phthalate (MnOP) and PAHs metabolite (1-hydroxypyrene, 1-OHP) were analyzed in spot urine samples collected from pregnant women. Correlation analysis and multiple regression analysis were conducted between the concentrations of maternal urinary metabolites and birth outcomes such as birth weight, birth length, head circumference and gestational age. Creatinine-corrected concentration (geometric mean; microg/g cre) was 9.14 (MMP), 9.76 (MEP), 51.6 (MnBP), 5.62 (MBzP), 5.45 (MEHP), 10.6 (MEHHP), 11.3 (MEOHP), 0.031 (MINP), 0.025 (MnOP) and 0.121 (1-OHP). These concentrations are comparable with literature value. The relationships between prenatal exposure to phthalate esters and birth outcomes were not significant. Statistically significant negative correlation was observed between 1-OHP and birth weight, birth length and head circumstances although the correlation was insignificant when only non-smokers were included in multiple regression analysis. In conclusion, we found that prenatal exposure to phthalate esters or PAHs did not affect birth outcomes at the exposure level of the present subjects.

Abstract  We have developed a gas chromatography-mass spectrometry (GC-MS) method to determine five phthalate monoesters (monoethyl phthalate (MEP), mono-n-butyl phthalate (MBP), mono-(2-ethylhexyl) phthalate (MEHP), monoisononyl phthalate (MINP) and monobenzyl phthalate (MBz)) in human urine. Human urine samples were subjected to enzymatic deconjugation of the glucuronides followed by extraction with hexane. The extracted phthalate monoesters were methylated with diazomethane, purified on a Florisil column and then subjected to GC-MS analysis. The recoveries from urine spiked with five phthalate monoesters were 86.3%-119% with coefficients of variation of 0.6%-6.1%. We measured phthalate monoester levels in human urine by analyzing 36 samples from volunteers. MBP and MEP were detected in all samples, and their median concentrations were 60.0 and 10.7 ng/mL, respectively. MBzP and MEHP were found in 75% and 56% of samples, and their median concentrations were 10.9 and 5.75 ng/mL, respectively. MINPs were not detected in most samples (6% detectable). Women had significantly (p < 0.05) higher mean concentrations of MBP and MEP than men. The estimated daily exposure levels for the four parent phthalates excluding diisononyl phthalate ranged from 0.27 to 5.69 mug/kg/day (median).

Abstract  We have developed a gas chromatography-mass spectrometry method to measure five phthalates (dibutyl phthalate, butylbenzyl phthalate, di-2-ethylhexyl phthalate, diisooctyl phthalate, and diisononyl phthalate) in diets and beddings for experimental animals. The recoveries from diets and beddings spiked with five phthalates were 98.8%-148% with coefficients of variation of 0.4%-7.8% for diets and 94.7%-146% with coefficients of variation of 1.0%-5.0% for beddings. We analyzed commercial animal diets and beddings, and found that the levels of phthalates varied from sample to sample; the concentrations of five phthalates were 141-1,410 ng/g for diets and 20.5-7,560 ng/g for beddings.

Abstract  Mono(2-ethylhexyl) phthalate (MEHP), the active metabolite of the testicular toxicant di(2-ethylhexyl) phthalate, inhibits FSH-stimulated rat Sertoli cell cAMP accumulation, stimulates basal lactate production, and decreases intracellular ATP levels in vitro. Dibutyl phthalate and dipentyl phthalate but not diethyldimethyl or dipropyl are also age-dependent testicular toxicants in vivo. We therefore examined the effect of animal age and phthalate monoester on the Sertoli cell FSH-stimulated cAMP accumulation, lactate secretion, and ATP levels in order to determine if these effects are part of the mechanism of action of phthalate esters in vivo. MEHP, monobutyl and monopentyl phthalates but not the monoethyl, monomethyl, or monopropyl phthalates inhibited FSH-stimulated cAMP accumulation, a segregation which matches the in vivo toxicity potential of these agents. MEHP and monopentyl, but not monobutyl phthalates, also stimulated Sertoli cell lactate secretion. The effect of the active phthalates on FSH-stimulated cAMP accumulation and lactate secretion is not dependent on age of animal over a range of 13-80 days, suggesting that the age-related toxicity in vivo may be related to differences in metabolism and disposition rather than tissue sensitivity. Since the ED50 of MEHP inhibition of cAMP accumulation and lactate secretion is similar, these two effects may be related to a common initial effect of the active phthalates. Inhibition of intracellular ATP levels is specific for MEHP and is lost with age (greater than 28 days of age) and thus is not likely to be an essential part of the in vivo mechanism of action of phthalate diesters.

Abstract  Phthalate esters are found in a wide variety of consumer and food packing products. Hence there is widespread exposure of the human population to these chemicals. Some of the phthalate esters are known to be toxic to the developing male reproductive system. This paper derives a reference dose (RfD) for each of the phthalate esters (dibutyl phthalate, diisobutyl phthalate, butylbenzyl phthalate, diethylhexyl phthalate, dipentyl phthalate, and diisononyl phthalate) that cause these effects. As these phthalate esters cause similar adverse biological effects and have the same mechanism of action, it is appropriate in a risk assessment to consider the potential adverse effects from cumulative exposure to these chemicals using a dose addition model. This paper provides examples of a cumulative risk assessment using the hazard index and relative potency approaches from the RfDs derived from studies in laboratory animals and exposure information in people. The results of the cumulative risk assessments for both a US and a German population show that the hazard index is below one. Thus it is unlikely that humans are suffering adverse developmental effects from current environmental exposure to these phthalate esters.

Abstract  BACKGROUND: Phthalates are widely used chemicals, and human exposure is extensive. Recent studies have indicated that phthalates may have thyroid-disrupting properties. OBJECTIVE: We aimed to assess concentrations of phthalate metabolites in urine samples from Danish children and to investigate the associations with thyroid function, insulin-like growth factor I (IGF-I), and growth. METHODS: In 845 children 4-9 years of age, we determined urinary concentrations of 12 phthalate metabolites and serum levels of thyroid-stimulating hormone, thyroid hormones, and IGF-I. RESULTS: Phthalate metabolites were detected in all urine samples, of which monobutyl phthalate was present in highest concentration. Phthalate metabolites were negatively associated with serum levels of free and total triiodothyronine, although statistically significant primarily in girls. Metabolites of di(2-ethylhexyl) phthalate and diisononyl phthalate were negatively associated with IGF-I in boys. Most phthalate metabolites were negatively associated with height, weight, body surface, and height gain in both sexes. CONCLUSIONS: Our study showed negative associations between urinary phthalate concentrations and thyroid hormones, IGF-I, and growth in children. Although our study was not designed to reveal the mechanism of action, the overall coherent negative associations between urine phthalate and thyroid and growth parameters may suggest causative negative roles of phthalate exposures for child health.

Abstract  BACKGROUND: Diisononyl phthalate (DINP), a principal plasticizer in many polyvinyl chloride products, has been shown to have an adjuvant effect on immunoglobulin (Ig) production in mice. However, the effects of DINP on allergic diseases have not been fully elucidated. OBJECTIVES: In the present study we investigated the effects of DINP on atopic dermatitis (AD)-like skin lesions induced by Dermatophagoides pteronyssinus (Dp) in atopic-prone NC/Nga mice. METHODS: Mice were injected intradermally with Dp on their ears and were exposed to DINP (0, 0.15, 1.5, 15, or 150 mg/kg/day) intraperitoneally. We evaluated clinical scores, ear thickening, histologic findings, protein expression of cytokines/chemokines in the ear, and serum levels of Ig and histamine. Furthermore, we investigated the effects of DINP on bone-marrow-derived dendritic cells (BMDCs) or splenocytes in vitro. After exposure to DINP (0-100 microM), cells were evaluated for phenotype and function. RESULTS: DINP aggravated AD-like skin lesions related to Dp. The aggravation was consistent with eosinophilic inflammation, mast cell degranulation, and thymic stromal lymphopoietin (TSLP) expression in the ear. DINP enhanced the expression of cell surface activation markers on BMDCs and their production of TARC/CCL17 (thymus- and activation-regulated chemokine) and MDC/CCL22 (macrophage-derived chemokine), as well as their capacity to stimulate Dp-specific T-cell proliferation. DINP also enhanced interleukin-4 production and Dp-stimulated proliferation of splenocytes. CONCLUSIONS: DINP can aggravate AD-like skin lesions related to Dp. The mechanisms of the aggravation might be mediated, at least partly, through the TSLP-related activation of dendritic cells and by direct or indirect activation of the immune cells.

Abstract  Eight phthalate esters, with alcohol chain lengths of 1-11 carbon atoms and with various degrees of branching, were tested in vitro in the L5178Y mouse lymphoma mammalian cell mutation assay and in the Balb/3T3 cell transformation assay. The tests were performed as part of a voluntary testing agreement between the Chemical Manufacturers Association's Phthalate Esters Panel and the United States Environmental Protection Agency (US EPA). The esters tested were: dimethyl phthalate (DMP), di-n-butyl phthalate (DBP), butyl benzyl phthalate (BBP), di- inverted question markn-hexyl, n-octyl, n-decyl inverted question mark phthalate (610P), di-isononyl phthalate (DINP), di- inverted question markheptyl, nonyl, undecyl inverted question mark phthalate (711P), di-isodecyl phthalate (DIDP) and di-undecyl phthalate (DUP). Both DMP and DBP were found to produce significant increases in the mutant frequency in the mouse lymphoma assay in the presence but not in the absence of an Aroclor-induced rat liver activation system (S-9). Ester 610P gave equivocal results in the mouse lymphoma assay in the presence and absence of rat liver S-9. There was no indication of mutagenic potential for any of the other test materials in the mouse lymphoma assay, and none of the test materials increased transformation frequency in the Balb/3T3 cell transformation assay. Aldehyde metabolites of the de-esterified alcohols are postulated to play a role in the positive results for DMP and DBP.

Abstract  A large number of phthalate esters were screened for estrogenic activity using a recombinant yeast screen. a selection of these was also tested for mitogenic effect on estrogen-responsive human breast cancer cells. A small number of the commercially available phthalates tested showed extremely weak estrogenic activity. The relative potencies of these descended in the order butyl benzyl phthalate (BBP) > dibutyl phthalate (DBP) > diisobutyl phthalate (DIBP) > diethyl phthalate (DEP) > diisiononyl phthalate (DINP). Potencies ranged from approximately 1 x 10(6) to 5 x 10(7) times less than 17beta-estradiol. The phthalates that were estrogenic in the yeast screen were also mitogenic on the human breast cancer cells. Di(2-ethylhexyl) phthalate (DEHP) showed no estrogenic activity in these in vitro assays. A number of metabolites were tested, including mono-butyl phthalate, mono-benzyl phthalate, mono-ethylhexyl phthalate, mon-n-octyl phthalate; all were wound to be inactive. One of the phthalates, ditridecyl phthalate (DTDP), produced inconsistent results; one sample was weakly estrogenic, whereas another, obtained from a different source, was inactive. analysis by gel chromatography-mass spectometry showed that the preparation exhibiting estrogenic activity contained 0.5% of the ortho-isomer of bisphenol A. It is likely that the presence of this antioxidant in the phthalate standard was responsible for the generation of a dose-response curve--which was not observed with an alternative sample that had not been supplemented with o,p'-bisphenol A--in the yeast screen; hence, DTDP is probably not weakly estrogenic. The activities of simple mixtures of BBP, DBP, and 17beta-estradiol were assessed in the yeast screen. No synergism was observed, although the activities of the mixtures were approximately additive. In summary, a small number of phthalates are weakly estrogenic in vitro. No data has yet been published on whether these are also estrogenic in vitro. No data has yet been published on whether these are also estrogenic in vivo; this will require tests using different classes of vertebrates and different routes of exposure.