Eucalyptol relieves the toxicity of diisobutyl phthalate in Ctenopharyngodon idellus kidney cells through Keap1/Nrf2/HO-1 pathway: Apoptosis-autophagy crosstalk and immunoregulation.

Diisobutyl phthalate (DiBP), one of the commonly used plasticizers in industry, is an endocrine disruptor and environmental contaminant that can persist in water and threaten the health of aquatic creatures. Eucalyptol (Euc), a monoterpenoid extracted from plants, has been proved to have anti-inflammatory, antioxidant, and detoxification properties. However, the protective mechanism of Euc against cell injury caused by DiBP exposure and the involvement of apoptosis, autophagy, and immunity remains unknown. In the current investigation, 27.8 µg/mL DiBP or/and 20 µM Euc has been applied to Ctenopharyngodon idellus kidney (CIK) cells for 24 h. The findings showed that exposure to DiBP raised intracellular ROS levels, inducing oxidative stress, and enhanced the rate of apoptosis as well as the expression of the apoptotic markers Bax, Caspase3, Caspase9, and Cytc while decreasing the expression of Bcl-2. Furthermore, DiBP inhibited IL-2, IFN-γ, Hepcidin-1, and ß-defensin expression and elevated TNF-α, and IL-1ß levels, causing immune dysfunction. DiBP and Euc co-treatment significantly activated the Keap1/Nrf2/HO-1 pathway, restored antioxidant enzyme activity, and elevated autophagy pathway-associated genes ATG5, Beclin1, and LC3B decreased p62 expression, enhanced cell autophagy, reduced apoptosis, and improved immunity. In conclusion, Euc promotes autophagy, alleviates DiBP-induced apoptosis, and improves immunological dysfunction in CIK cells by regulating the Keap1/Nrf2/HO-1 pathway. These results demonstrated the threat of DiBP exposure to fish while providing a theoretical foundation for using Euc in aquaculture.

Development of a whole-cell biocatalyst for diisobutyl phthalate degradation by functional display of a carboxylesterase on the surface of Escherichia coli.

BACKGROUND: Phthalic acid esters (PAEs) are widely used as plasticizers or additives during the industrial manufacturing of plastic products. PAEs have been detected in both aquatic and terrestrial environments due to their overuse. Exposure of PAEs results in human health concerns and environmental pollution. Diisobutyl phthalate is one of the main plasticizers in PAEs. Cell surface display of recombinant proteins has become a powerful tool for biotechnology applications. In this current study, a carboxylesterase was displayed on the surface of Escherichia coli cells, for use as whole-cell biocatalyst in diisobutyl phthalate biodegradation. RESULTS: A carboxylesterase-encoding gene (carEW) identified from Bacillus sp. K91, was fused to the N-terminal of ice nucleation protein (inpn) anchor from Pseudomonas syringae and gfp gene, and the fused protein was then cloned into pET-28a(+) vector and was expressed in Escherichia coli BL21(DE3) cells. The surface localization of INPN-CarEW/or INPN-CarEW-GFP fusion protein was confirmed by SDS-PAGE, western blot, proteinase accessibility assay, and green fluorescence measurement. The catalytic activity of the constructed E. coli surface-displayed cells was determined. The cell-surface-displayed CarEW displayed optimal temperature of 45 °C and optimal pH of 9.0, using p-NPC2 as substrate. In addition, the whole cell biocatalyst retained ~ 100% and ~ 200% of its original activity per OD600 over a period of 23 days at 45 °C and one month at 4 °C, exhibiting the better stability than free CarEW. Furthermore, approximately 1.5 mg/ml of DiBP was degraded by 10 U of surface-displayed CarEW cells in 120 min. CONCLUSIONS: This work provides a promising strategy of cost-efficient biodegradation of diisobutyl phthalate for environmental bioremediation by displaying CarEW on the surface of E. coli cells. This approach might also provide a reference in treatment of other different kinds of environmental pollutants by displaying the enzyme of interest on the cell surface of a harmless microorganism.

Presence, distribution and risk assessment of phthalic acid esters (PAEs) in suburban plastic film pepper-growing greenhouses with different service life.

The widespread usage of plastic film increased the content of phthalic acid esters (PAEs) in the environment, causing PAE residue in vegetables and subsequently increasing health risks to humans when consuming them. In this work, the presence, distribution and risk assessment of 15 PAEs in soils and peppers from suburban plastic film pepper-growing greenhouses were investigated. The total PAE contents in soil and pepper samples ranged from 320.1 to 971.2 µg/kg (586.3 µg/kg on average) and from 196.6 to 304.2 µg/kg (245.4 µg/kg on average), respectively. Di (2-ethyl)hexyl, dibutyl and diisobutyl phthalates (DEHP, DnBP and DiBP, respectively) were the most abundant in both soil and pepper samples. Specifically, DEHP showed the highest content in soils, while the DnBP content was the highest in peppers. The total PAE content in soils from pepper-greenhouses was much lower than in the agricultural soils mulched with plastic films, but significantly higher than in the agricultural soils from open uncovered fields. The total PAE content in peppers decreased as the service life of plastic film greenhouses increased. Correlation analysis suggested that the difference in distribution and accumulation behaviors of individual PAEs in greenhouse systems was correlated with their physicochemical properties. The non-cancer and carcinogenic risks of priority PAEs show low risks of PAEs detected in pepper and soil samples from the suburban plastic film greenhouses to human health.

Spatial distribution of phthalate esters and the associated response of enzyme activities and microbial community composition in typical plastic-shed vegetable soils in China.

The widespread use of phthalate esters (PAEs) in plastic products has made them ubiquitous in environment. In this study, 93 soil samples were collected in 31 plastic-sheds from one of China's largest vegetable production bases, Shouguang City, Shandong Province, to investigate the pollution characteristics and composition of PAEs in soils. Eleven PAEs were detected in the soil samples with the total concentration of 756-1590 µg kg-1 dry soil. Di (2-ethylhexyl) phthalate (DEHP), bis (2-n-butoxyethyl) phthalate (DBEP), di-isobutyl phthalate (DiBP) and di-n-butyl phthalate (DBP) were the main pollutants with the highest concentrations. Moreover, soil properties, including pH, total organic carbon (TOC), soil enzyme activities, and soil microbial community characteristics, were monitored to explore the associated formation mechanisms. The concentration of PAEs in the plastic-shed vegetable soils was regionalized and the contamination degree in different regions was related to soil microbial characteristics and soil enzyme activities. Phthalate ester is positively correlated with catalase and sucrase, and negatively correlated with dehydrogenase and urease. Furthermore, some tolerant and sensitive bacteria were selected, which possibly could be used as potential indicators of PAE contamination in soil. Dimethyl phthalate (DMP) and DBP also had greater effects on the soil microbial community than other PAEs. The results will provide essential data and support the control of PAEs in plastic-shed vegetable soils in China.

Acute effects on human sperm exposed in vitro to cadmium chloride and diisobutyl phthalate.

Epidemiological studies reported a negative relationship between concentrations of heavy metals and phthalates in seminal fluid and semen quality, likely compromising male fertility potential. The aim of this study was to investigate the in vitro effects of cadmium chloride (CdCl2), a common heavy metal, and diisobutyl phthalate (DIBP), a common phthalate ester, on human sperm functions necessary for fertilization. After in vitro incubation of spermatozoa with 10 µM CdCl2 or 100 and 200 µM DIBP for 24 h, a significant decrease of sperm progressive and hyperactivated motility was observed. The exposure to each of the two toxic agents also induced spontaneous sperm acrosome reaction and blunted the physiological response to progesterone. Both agents induced an increase of caspase activity suggesting triggering of an apoptotic pathway. Our results suggest that acute exposure of spermatozoa to these pollutants may impair sperm ability to reach and fertilize the oocyte.

Characterization and Identification of Recalcitrant Organic Pollutants (ROPs) in Tannery Wastewater and Its Phytotoxicity Evaluation for Environmental Safety.

Tannery wastewater (TWW) is of serious environmental concern to pollution control authorities, because it contains highly toxic, recalcitrant organic and inorganic pollutants. The nature and characteristics of recalcitrant organic pollutants (ROPs) are not fully explored to date. Hence, the purpose of this study was to characterize and identify the ROPs present in the treated TWW. Gas chromatography-mass spectrometry data analysis showed the presence of a variety of ROPs in the treated TWW. Results unfolded that benzyl chloride, butyl octyl phthalate, 2,6-dihydroxybenzoic acid 3TMS, dibutyl phthalate, benzyl alcohol, benzyl butyl phthalate, 4-chloro-3-methyl phenol, phthalic acid, 2'6'-dihydroxyacetophenone, diisobutyl phthalate, 4-biphenyltrimethylsiloxane, di-(-2ethy hexyl)phthalate, 1,2-benzenedicarboxylic acid, dibenzyl phthalate, and nonylphenol were present in the treated TWW. Due to endocrine disrupting nature and aquatic toxicity, the U.S. Environmental Protection Agency classified many of these as "priority pollutants" and restricted their use in leather industries. In addition, the physicochemical analysis of the treated TWW also showed very high BOD, COD, and TDS values along with high Cr and Pb content beyond the permissible limits for industrial discharge. Furthermore, phytotoxicity assessment unfolds the inhibitory effects of TWW on the seed germination, seedling growth parameters, and α-amylase activity in Phaseolus aureus L. This indicates that the TWW discharged even after secondary treatment into the environment has very high pollution parameters and may cause a variety of serious health threats in living beings upon exposure. Overall, the results reported in this study will be helpful for the proper treatment and management of TWW to combat the environmental threats.

SPME-Based Ca-History Method for Measuring SVOC Diffusion Coefficients in Clothing Material.

Clothes play an important role in dermal exposure to indoor semivolatile organic compounds (SVOCs). The diffusion coefficient of SVOCs in clothing material (Dm) is essential for estimating SVOC sorption by clothing material and subsequent dermal exposure to SVOCs. However, few studies have reported the measured Dm for clothing materials. In this paper, we present the solid-phase microextraction (SPME) based Ca-history method. To the best of our knowledge, this is the first try to measure Dm with known relative standard deviation (RSD). A thin sealed chamber is formed by a circular ring and two pieces of flat SVOC source materials that are tightly covered by the targeted clothing materials. Dm is obtained by applying an SVOC mass transfer model in the chamber to the history of gas-phase SVOC concentrations (Ca) in the chamber measured by SPME. Dm's of three SVOCs, di-iso-butyl phthalate (DiBP), di-n-butyl phthalate (DnBP), and tris(1-chloro-2-propyl) phosphate (TCPP), in a cotton T-shirt can be obtained within 16 days, with RSD less than 3%. This study should prove useful for measuring SVOC Dm in various sink materials. Further studies are expected to facilitate application of this method and investigate the effects of temperature, relative humidity, and clothing material on Dm.

Semi-volatile organic compounds in the air and dust of 30 French schools: a pilot study.

The contamination of indoor environments with chemical compounds released by materials and furniture, such as semi-volatile organic compounds (SVOCs), is less documented in schools than in dwellings-yet children spend 16% of their time in schools, where they can also be exposed. This study is one of the first to describe the contamination of the air and dust of 90 classrooms from 30 nursery and primary schools by 55 SVOCs, including pesticides, phosphoric esters, musks, polycyclic aromatic hydrocarbons (PAHs), polychlorobiphenyls (PCBs), phthalates, and polybromodiphenylethers (PBDEs). Air samples were collected using an active sampling method, and dust samples were collected via two sampling methods (wiping and vacuum cleaning). In air, the highest concentrations (median >100 ng/m3 ) were measured for diisobutyl phthalate (DiBP), dibutyl phthalate (DBP), diethyl phthalate (DEP), bis(2-ethylhexyl) phthalate (DEHP), and galaxolide. In dust, the highest concentrations (median >30 µg/g) were found for DEHP, diisononyl phthalate (DiNP), DiBP, and DBP. An attempt to compare two floor dust sampling methods using a single unit (ng/m²) was carried out. SVOC concentrations were higher in wiped dust, but frequencies of quantification were greater in vacuumed dust.

Phthalate induced toxicity in prostate cancer cell lines and effects of alpha lipoic acid.

OBJECTIVE: The effects of dimethyl phthalate, diethyl phthalate, diisobutyl phthalate, di-n-butyl phthalate, benzylbutyl phthalate, di-2-ethylhexyl phthalate were investigated on human prostate cancer cell lines DU145 and PC3 in vitro. MATERIALS AND METHODS: Standards of dimethyl phthalate, diethyl phthalate, di-isobutyl phthalate, dibutyl phthalate, benzyl butyl phthalate, and di-ethyl hexyl phthalate were used. Alpha lipoic acid was used as antioxidant compound. DU145 and PC3 human prostate carcinoma cells were used. MTT assay were used for cytotoxicity assay. RESULTS: A low dose proliferative effect of phthalates in vitro was observed. With the hypothesis of the inhibition of aerobic glycolysis activity in cancer treatment, α-lipoic acid was applied to cells; where as a contrary to previous studies, no change in the cell proliferation was observed. In combination with ALA, at IC50 and lower doses, an increase of the cytotoxic effect was found for DIBP, DBP and BBP; while for DMP, DEP and DEHP, a decrease was observed for DU145 cells. In PC3 cells, a decrease was observed for DMP, DEP and DBPs; while no significant difference were observed for DEHP, DIBP and BBP. CONCLUSSION: The present study demonstrates preliminary information regarding the low dose proliferative effects of phthalates in prostate cancer in vitro (Tab. 2, Fig. 2, Ref. 65).

Dietary exposure to di-isobutyl phthalate increases urinary 5-methyl-2'-deoxycytidine level and affects reproductive function in adult male mice.

Phthalates are a large family of ubiquitous environmental pollutants suspected of being endocrine disruptors. Epidemiological studies have associated phthalate metabolites with decreased reproductive parameters and linked phthalate exposure with the level of urinary 5-methyl-2'-deoxycytidine (5mdC, a product of methylated DNA). In this study, adult male mice were exposed to 450mg di-isobutyl phthalate (DiBP)/(kg·day) via dietary exposure for 28days. Mono-isobutyl phthalate (MiBP, the urinary metabolite) and reproductive function parameters were determined. The levels of 5mdC and 5-hydroxymethyl-2'-deoxycytidine (5hmdC) were measured in urine to evaluate if their contents were also altered by DiBP exposure in this animal model. Results showed that DiBP exposure led to a significant increase in the urinary 5mdC level and significant decreases in sperm concentration and motility in the epididymis, accompanied with reduced testosterone levels and down-regulation of the P450 cholesterol side-chain cleavage enzyme (P450scc) gene in the mice testes. Our findings indicated that exposure to DiBP increased the urinary 5mdC levels, which supported our recent epidemiological study about the associations of urinary 5mdC with phthalate exposure in the male human population. In addition, DiBP exposure impaired male reproductive function, possibly by disturbing testosterone levels; P450scc might be a major steroidogenic enzyme targeted by DiBP or other phthalates.

An integrated exposure assessment of phthalates for the general population in China based on both exposure scenario and biomonitoring estimation approaches.

The representativeness of available studies on integrated exposure assessment of phthalates for the general population in China is lacking. Based on an exhaustive review of the extensive monitoring data available for China, this study presents a large-scale estimation of exposure levels to three typical phthalates, di(2-ethylhexyl) phthalate (DEHP), di-n-butyl phthalate (DBP) and diisobutyl phthalate (DiBP), by applying both exposure scenario and biomonitoring estimation approaches. The respective median exposure levels from the exposure scenario and biomonitoring estimation approaches were 3.80, 3.02 and 1.00 µg/kg bw/day and 3.38, 3.21 and 3.32 µg/kg bw/day for DEHP, DBP and DiBP, which are acceptable levels of exposure with respect to current international guidelines. Evaluation results from the two approaches showed both similarities and differences among the different phthalates, making the exposure assessment comparable and more comprehensive. In terms of sources of exposure, food intake was the largest contributor, while indoor air exposure had greater contribution to the estimated daily intakes (EDIs) of DiBP than that of the other phthalates. Moreover, more attention should be paid to the higher exposure levels of phthalates in several intensively industrialized and urbanized areas, and the causes of the different exposure levels in the different regions need to be further explored.

Urinary excretion of phthalate metabolites in school children of China: implication for cumulative risk assessment of phthalate exposure.

We analyzed 13 metabolites of 9 phthalates in urine of 782 Chinese school children aged 8­11 years and estimated the daily intake for phthalates based on urinary metabolite levels. The daily intakes were compared with acceptable intake levels to calculate the hazard quotient (HQ) for single phthalate. Finally, the cumulative risk for each child was assessed by means of a hazard index (HI) which is the sum of HQs. Overall, 11 metabolites were found in at least 85% of the urine samples with the highest median concentration of 47.1 ng/mL (93.4 µg/g creatinine) for mono-n-butyl phthalate (MnBP). Monooctyl phthalate (MOP) and monoisononyl phthalate (MiNP) were not detectable. The cumulative risk assessment covering di(2-ethylhexyl) phthalate (DEHP), di-n-butyl phthalate (DnBP), di-isobutyl phthalate (DiBP), and butyl-benzyl phthalate (BBzP) demonstrated that 19.8% (volume model-based) and 40.3% (creatinine model-based) of the children exceeded 1 for the HI based on tolerable daily intake (TDI) values (considered as potential adverse antiandrogenic effect). Furthermore, at least 36% of the children from the manufacturing-intensive region had a HI higher than 1. The results indicate that Chinese children are widely exposed to phthalates and those from manufacturing-intensive regions are probably at a high risk of cumulative phthalate exposure.

Comparison, association, and risk assessment of phthalates in floor dust at different indoor environments in Delaware, USA.

This study aimed to compare and assess phthalate contamination in various indoor environments. In this study, 44 floor dust samples from different indoor environments in Delaware, USA were collected and analyzed for 14 phthalates using gas chromatography-mass spectrometry. Phthalates were detected in all dust samples with the total concentration ranging from 84 to 7117 mg kg(-1). DEHP (di-2-ethylhexyl phthalate), BzBP (benzylbutyl phthalate), DBP (dibutyl phthalate), and DiBP (di-isobutyl phthalate) were both the most frequently and abundantly detected phthalates. The average concentration of total phthalates in dust from offices, student dorms, gyms, stores, and daycare centers was found to be significantly or insignificantly (P = 0.05) higher than that in dust from houses and apartments. Plastic flooring materials and the application of floor care chemical products were positively associated with total phthalate concentration in floor dust. Toxicological risk assessment indicated that an investigated daycare center in this study was the only indoor environment that may cause the intake amount of DEHP of infants, toddlers, and children via dust ingestion to exceed the reference dose established by the U.S. Environmental Protection Agency (USEPA). Regular monitoring on phthalate contamination in sensitive indoor environments is recommended.

Estimated daily intake and hazard quotients and indices of phthtalate diesters for young danish men.

Because of wide exposure to phthalates, we investigated whether simultaneous exposure to several phthalates reached levels that might cause adverse antiandrogenic effects. Thirty three healthy young Danish men each delivered three 24-h urine samples during a three months period. The daily intakes of the sum of di-n-butyl and di-iso-butyl phthalate, di(2-ethylhexyl) phthalate, di-iso-nonyl phthalate, and butylbenzyl phthalate were estimated based on urinary excretion of the metabolites. Based on a hazard quotient (HQ) of the individual phthalate (i.e., the ratio between the daily intake and an acceptable level of exposure), a hazard index (HI) for each man was calculated as the sum of HQs for the individual phthalates. All men were exposed to all phthalates during the urine collection periods. Median HIs were all below 1 (i.e., below an acceptable cumulative threshold) ranging from 0.11 to 0.17 over the three different sample collections. Of the 33 men, 2 men had HIs above 1 in one of their three samples, indicating that occasionally the combined exposure to the investigated phthalates reached a level that may not be considered safe. Besides the phthalates investigated here, humans are exposed to numerous other chemicals that also may contribute to a cumulative antiandrogenic exposure.

[A new phthalide from angelicae sinensis radix].

Angelicae Sinensis Radix, dried roots of Angelicae sinensis, is a well-known traditional Chinese medicine. Six compounds were isolated and purified by silica gel and preparative HPLC. According to physicochemical properties and spectral data, the compounds were identified as senkyunolide H-7-acetate (1), o-phthalic acid (2), diisobutyl phthalate (3), p-hydroxyphenylethanol ferulate (4), ferulic acid (5) and coniferylferulate (6). Compound 1 was a new one.

Developmental toxicity and mechanism of dibutyl phthalate and alternative diisobutyl phthalate in the early life stages of zebrafish (Danio rerio).

Diisobutyl phthalate (DiBP), is widely chemical replacement for Dibutyl phthalate (DBP). Although DBP and DiBP have been detected in surface water worldwide, few studies to date have systematically assessed the risks of DBP and its alternatives to aquatic organisms. The present study compared DBP and DiBP for their individual and joint toxicity as well as thyroid hormone levels in zebrafish embryo. Transcripts of key genes related to the hypothalamic-pituitary-thyroid (HPT) axis were investigated in developing zebrafish larvae by application of real time polymerase chain reaction. The median half-lethal concentrations of DBP and DiBP to zebrafish at 96 h were 0.545 mg L-1 and 1.149 mg L-1, respectively. The joint toxic effect of DBP-DiBP (0.25-0.53 mg L-1) with the same ratio showed a synergistic effect. Thyroid hormones levels increased with exposure to 10 µg L-1 of DBP or 50 µg L-1 of DiBP, and exposure to both compounds significantly increased thyroid gland-specific transcription of thyroglobulin gene (tg), hyronine deiodinase (dio2), and transthyretin (ttr), indicating an adverse effect associated with the HPT axis. Molecular docking results indicated that DBP (-7.10 kcal/M and -7.53 kcal/M) and DiBP (-6.63 kcal/M and -7.42 kcal/M) had the same docking energy with thyroid hormone receptors. Our data facilities an understand of potential harmful effects of DBP and its alternative (DiBP).

[Effects of DiBP on the cAMP/PKA-CREB-BDNF signaling pathway of hippocampus in mice].

OBJECTIVE: To explore the effects of diisobutyl phthalate (DiBP) on the cAMP/PKA-CREB signaling pathway of hippocampus in mice. METHODS: Accommodating 30 KunMing mice to the animal house for 3 days, then dividing the mice into 5 groups according to their weights. That is, one control group and four experimental groups (I group, 50 mg/kg BW; II group, 250 mg/kg BW; Ill group, 500 mg/kg BW; IV group, 1000 mg/kg BW). The mice were fed with the corn oil in control group, and the other groups were fed with the related dose of DiBP mixture by gavages last for 8 weeks. At the end of experimental time, the mice were killed, and the tissue samples of hippocampus were taken immediately. The content of cAMP was determined by ELISA, and p-CREB, P-PKA C were measured by western blotting, while mRNA expressions including CREB, BDNF, c-fos and c-jun were checked by RT-PCR. RESULTS: The cAMP content and the p-PKA C protein of hippocampus in IV group was significantly less than control group (P<0.05) and compared with control group, the p-CREB protein of hippocampus in 1I group decreased (P<0.05), while the relative level of CREB, BDNF, c-fos and c-jun mRNA were down-regulated in all experimental groups. CONCLUSIONS: The abnormal changes of expression of the signal molecules composing the cAMP/PKA-CREB signaling pathway were observed in the hippocampus of DiBP exposure mice. This might be one of the possible mechanisms of DiBP induced cognitive impairment.

[Effect of diisobutyl phthalate on learning and memory behavior and apoptosis of hippocampus cells in mice].

OBJECTIVE: To give the original research of diisobutyl phthalate (DiBP) on learning and memory behavior, determine whether it can through blood-brain barrier and effect apoptosis of hippocampus cells in mice. METHODS: Accommodating 60 Kunming mice to the animal house for 3 days, then dividing the mice into 5 groups according to their weights. That is, one control group and four experimental groups (I group, 50 mg/kg BW. II group, 250mg/kg BW. III group, 500 mg/kg BW. IV group, 1000 mg/kg BW). The mice were fed with the corn oil in control group, and the other groups were fed with the related dose of diisobutyl phthalate mixture by gavages last for 8 weeks. At the end of experimental time, passive avoidance response was examined, then all of mice were killed, and choosed the brain tissues to test the DiBP content and apoptosis rate of hippocampal cells and hippocampal ultrastructural alterations on electron microscopy. RESULTS: In the passive avoidance response test, the exposed animals of IV group showed learning impairment as compared to unexposed mice (P < 0.05). DiBP was detected in III group and IV group, the mean content of them were (1.27 +/- 0.56) and (1.96 +/- 0.42) microg/g. The apoptosis rate of hippocampal cells (IV group vs control group) increase significantly (P < 0.05). Hippocampal ultrastructural were damaged in all dose-groups. CONCLUSION: As a result, in the experiments, exposure to DiBP could exert passive avoidance neurobehavioral effects. DiBP could through blood-brain barrier after oral intake, and disordered the way of apoptosis of hippocampal cells, and morphologic change of mitochondria mybe is the main reason of changes of neuron apoptosis.

In situ ATR-FTIR studies on MgCl2-diisobutyl phthalate interactions in thin film Ziegler-Natta catalysts.

To study the surface structure of MgCl(2) support and its interaction with other active components in Ziegler-Natta catalyst, such as electron donors, we prepared a thin film analogue for Ziegler-Natta ethylene polymerization catalyst support by spin-coating a solution of MgCl(2) in ethanol, optionally containing a diester internal donor (diisobutyl-ortho-phthalate, DIBP) on a flat Si crystal surface. The donor content of these films was quantified by applying attenuated total internal reflection-Fourier transform infrared spectroscopy (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS). Changes in the interaction of DIBP with MgCl(2) at various temperatures were monitored by in situ ATR-FTIR. Upon increasing the temperature, a shift in the (C═O) band toward lower wavenumbers was observed together with the depletion of (O-H) stretching band due to the desorption of residual ethanol. We assign this shift to gradual redistribution of adsorbed DIBP from adsorption sites on the MgCl(2) (104) surface toward the more acidic MgCl(2) (110) surface. The morphologies of MgCl(2) and MgCl(2)/DIBP films were studied by transmission electron microscopy (TEM) revealing a preferential orientation of ClMgCl layers (001) parallel to the lateral film dimensions. This orientation becomes more pronounced upon annealing. In the absence of donor, the MgCl(2) grow in to large crystals aligned in large domains upon annealing. Both crystal growth and alignment is impeded by the presence of donor.

Di-n-butyl phthalate (DnBP) and diisobutyl phthalate (DiBP) metabolism in a human volunteer after single oral doses.

An individual (male, 36 years, 87 kg) ingested two separate doses of di-n-butyl phthalate (DnBP) and diisobutyl phthalate (DiBP) at a rate of ~60 µg/kg. Key monoester and oxidized metabolites were identified and quantified in urine continuously collected until 48 h post-dose. For both DnBP and DiBP, the majority of the dose was excreted in the first 24 h (92.2 % of DnBP, 90.3 % of DiBP), while only <1 % of the dose was excreted in urine on day 2. In each case, the simple monoesters were the major metabolites (MnBP, 84 %; MiBP, 71 %). For DnBP, ~8 % was excreted as various side chain oxidized metabolites. For DiBP, approximately 20 % was excreted mainly as the oxidized side chain metabolite 2OH-MiBP, indicating that the extent of oxidative modification is around 2.5 times higher for DiBP than for DnBP. All DnBP and DiBP metabolites reached peak concentrations between 2 and 4 h post-exposure, followed by a monotonic decline. For DnBP metabolites, the elimination halftime of MnBP was 2.6 h; longer elimination halftimes were estimated for the oxidized metabolites (2.9-6.9 h). For DiBP metabolites, MiBP had the shortest halftime (3.9 h), and the oxidized metabolites had somewhat longer halftimes (4.1 and 4.2 h). Together with the simple monoesters, secondary oxidized metabolites are additional and valuable biomarkers of phthalate exposure. This study provides basic human metabolism and toxicokinetic data for two phthalates that have to be considered human reproductive toxicants and that have been shown to be omnipresent in humans.