Glycine-β-cyclodextrin-assisted cometabolism of phenanthrene and pyrene by Pseudomonas stutzeri DJP 1 from marine sediment

Cometabolic degradation is currently an effective and extensively way to remove high molecular weight polycyclic aromatic hydrocarbons(HMW-PAHs).Unfortunately,due to low bio-accessibility and high biotoxicity,the cometabolic degradation rate of HMW-PAHs is limited.Glycine-β-cyclodextrin(GCD)was obtained through amino modification ofβ-cyclodextrin(BCD)and added to cometabolic system of phenanthrene(PHE)and pyrene(PYR)to assist PYR biodegradation.Results show that the addition of GCD(100 mg/L)effectively improved the removal rate of PYR(20 mg/L)by 42.3%.GCD appeared to increase the bio-accessibility and reduce the biotoxicity of PHE and PYR,and then promoted the growth of Pseudomonas stutzeri DJP1 and stimulated the elevation of dehydrogenase(DHA)and catechol 12 dioxygenase(C12O)activities.The phthalate metabolic pathway was accelerated,which improved the cometabolic degradation.This study provided a new reference for the cometabolic degradation of HMW-PAHs.

Determination of Benzo[a]pyrene in Edible Oil by High Performance Liquid Chromatography-Fluorescence Detector (HPLC-FL)

In this study, an optimized high performance liquid chromatography-fluorescence detector (HPLC-FL) method for the determination of benzo[a]pyrene in edible oil was established. HPLC was performed with Thermo Fisher Scientific C18 column (250 mm×4.6 mm, 5 μm) as the chromatographic column and acetonitrile and water as the mobile phase, and the excitation wavelength and emission wavelength of fluorescence detector were 286 and 430 nm, respectively. The response was high, and the linear range was 0.5-10.0 ng/ml. The lowest limit of detection was 0.11 ng/ml, and the average recovery was 92.5%. This method is suitable for quantitative analysis of benzo[a]pyrene content in edible oil.

Switch of phosphorylation to O-GlcNAcylation of AhR contributes to vascular oxidative stress induced by benzo[a]pyrene

Benzo[a]pyrene(B[a]P)is a food contaminant toxic for cardiovascular diseases.The nuclear translocation of Arylhydrocarbon receptor(AhR)plays an important role in B[a]P-induced oxidative stress and vascular diseases.We confi rmed that B[a]P promoted ROS production in vascular smooth muscle cells(VSMCs)in vitro and in vivo,associated with the nuclear translocation of AhR.It is known that phosphorylation inhibits while dephosphorylation of AhR promotes nuclear translocation of AhR.However,from the posttranslational modifi cation level,the mechanism by which B[a]P activates and regulates the nuclear translocation of AhR is unclear.Co-immunoprecipitation results showed that cytoplasmic AhR was phosphorylated before B[a]P stimulation,and switched to O-GlcNAcylation upon B[a]P 1-h stimulation in VSMCs,suggesting there may be a competitively inhibitory relationship between O-GlcNAcylation and phosphorylation of AhR.Next,siRNAs of O-linked N-acetylglucosamine transferase(OGT),O-GlcNAcase(OGA)and OGA inhibitor PUGNAc were used.SiOGT blocks but siOGA and PUGNAc promote B[a]P-dependent AhR nuclear translocation and oxidative stress.Ser11 may be the competitive binding site for phosphorylation and O-GlcNAcylation of AhR.Phosphorylation-mimic variant inhibits but O-GlcNAcylation of AhR promotes AhR nuclear translocation and oxidative stress.Our fi ndings highlight a new perspective for AhR nuclear translocation regulated by the competitive modifi cation between phosphorylation and O-GlcNAcylation.

Pyrene分子(小片石墨晶体)的电子传导特性(英文)

利用量子理论中基于Green函数的tight-binding方法,对pyrene分子的电子传导和电子流分布进行了理论研究。在考虑到界面耦合和Hopping积分的情况下,得出了电子透射谱和流分布的模拟结果。结果显示透射与电子的能量紧密相关;谱的振荡特征是能级量子化的结果;流分布有着特定的方向,并且在每一个原子点上符合Kirchhoff量子流守恒定律。另外还发现了桥接pyrene分子的正负能开关特性。

Pyrene衍生物掺杂SiO_2荧光纳米粒子的制备及光物理性能

以泊洛沙姆(F-127)为模板剂,以硅氧烷化芘的衍生物4-(1-芘基)-N-[3-(三乙氧基硅基)丙基]丁酰胺(Pyrene-1)与正硅酸四乙酯(TEOS)的混合物为硅源,通过胶束模板法制备了Pyrene-1掺杂的SiO_2荧光纳米粒子.采用透射电子显微镜(TEM)、动态光散射仪(DLS)、紫外-可见分光光度计和荧光光谱仪等对Pyrene-1掺杂的SiO_2荧光纳米粒子的表观形貌、粒径分布和光物理性质进行了表征.结果表明,所制备Pyrene-1掺杂的SiO_2荧光纳米粒子粒径均一(TEM统计粒径约为12 nm,DLS表征的水合直径约为33 nm),在水中具有很好的分散稳定性.通过调节染料的掺杂比例可以有效调控染料分子间距.同时,由于染料分子被共价键固定在Si—O—Si骨架中,使Pyrene-1掺杂的SiO_2荧光纳米粒子的荧光量子效率大幅提高(从有机溶剂中的0.15提升至0.95),荧光寿命得以延长(从有机溶剂中的17.9 ns延长至191.8 ns).在染料掺杂比例较高时,通过生成激基缔合物(Excimer),形成了具有较大Stokes位移(~100 nm)的发光.

Inhibitory effects of RRR-α-tocopheryl succinate on benzo (a) pyrene (B (a) P)-induced forestomach carcinogenesis in female mice

AIM To study the inhibitory effects of VES( RRR-α-tocopheryl Succinate, VES ), aderivative of natural Vitamin E, on benzo (a)pyrene (B (a) P)-induced forestomach tumor infemale mice.METHODS The model of B (a)P-inducedforestomach tumor was established according tothe methods of Wattenberg with slightmodifications. One hundred and eighty femalemice (6 weeks old) were divided into six groupsequally; negative control (Succinic acid),vehicle control ( Succinate + B (a) P), positivecontrol(B(a) P), high VES(2.5g/kg. b. w + B(a)P), Iow VES(1 .25 g/kg. b. w + B(a) P) ig as wellas VES by ip (20 mg/kg, b. w + B(a) P). Exceptthe negative control group, the mice wereadministrated with B(a)P ig. and correspondingtreatments for 4 weeks to study the anti-carcinogenetic effect of VES during the initiationperiod. The experiment lasted 29 weeks, inwhich the inhibitory effects of VES both ontumor incidence and tumor size were tested.RESULTS The models of B (a)P-inducedforestomach tumor in female mice wereestablished successfully. Some werecauliflower-like, others looked like papilla, evena few were formed into the ulcer cavities. VES at1.25 g/kg. b. w, 2.5 g/kg. b.w. by ig and 20 mg/kg. b. w. via ip could decrease the number oftumors per mouse (1.7 ± 0. 41, 1.6 ± 0.34 and 1.1±0.43), being lower than that of B(a)P group(5.4 ± 0.32, P＜0.05). The tumor incidence wasinhibited by 18.2%, 23.1% and 50.0%. VES at1.25g/kg.b.w., 2.5 g/ kg.b.w. by ig and20 mg/kg. b.w. via ip reduced the total volumeof tumors per mouse (54.8 ± 8.84, 28.4 ± 8.32and 23.9± 16.05), being significantly lower thanthat of B(a)P group (150.2±20.93, P＜0.01).The inhibitory rates were 63.5%, 81.1% and84.1%, respectively.CONCLUSION VES has inhibitory effects on B(a) P-induced forestomach carcinogenesis infemale mice, especially by ip and it may be apotential anti-cancer agent in vivo.

Phytoremediation for phenanthrene and pyrene contaminated soils

Phytoremediation of soil contaminated with phenanthrene and pyrene was investigated using twelve plant species. Plant uptake and accumulation of these chemicals were evaluated. At the end of the experiment(45 d), the remaining respective concentrations of soil phenanthrene and pyrene in spiked vegetated soils, with initial phenanthrene of 133 3 mg/kg and pyrene of 171 5 mg/kg, were 8 71—16 4 and 44 9—65 0 mg/kg, generally 4 7%—49 4% and 7 1%—35 9% lower than their concentrations in the non vegetated soils. The loss of phenanthrene and pyrene in vegetated spiked soils were 88 2%—93 0% and 62 3%—73 8% of the added amounts of these contaminants, respectively. Although plant uptake and accumulation of these compounds were evident, and root concentrations and RCFs(root concentration factors; defined as the ratio of PAH concentrations in roots and in the soils on a dry weight basis) of these compounds significantly positively correlated to root lipid contents, plant uptake and accumulation only accounted for less than 0 01% and 0 23% of the enhanced loss of these chemicals in vegetated versus non vegetated soils. In contrast, plant promoted microbial biodegradation was the dominant mechanism of the phytoremediation for soil phenanthrene and pyrene contamination. Results from this study suggested a feasibility of the establishment of phytoremediation for soil PAH contamination.

Photochemical behavior of benzo[a]pyrene on soil surfaces under UV light irradiation

The rates of photodegradation and photocatalysis of benzo [a]pyrene （BaP） on soil surfaces under UV light have been studied. Different parameters such as temperature, soil particle sizes, and soil depth responsible for photodegradation, catalyst loads and wavelength of UV irradiation blamed for photocatalysis have been monitored. The results obtained indicated that BaP photodegradation follows pseudo-first-order kinetics. BaP photodegradation was the fastest at 30℃ . The rates of BaP photodegradation at different soil particle size followed the order： less than 1 mm〉less than 0.45 mm〉less than 0.25 mm. When the soil depth increased from 1 mm to 4 ram, the half-life increased from 13.23 d to 17.73 d. The additions of TiO2 or Fe2O3 accelerated the photodegradation of BaP, and the photocatalysis of BaP follows pseudo-first-order kinetics. Changes in catalyst loads of TiO2 （0.5%, 1%, 2%, and 3% （wt）） or Fe203 （2%, 5%, 7%, and 10% （wt）） did not significantly affect the degradation rates. Both BaP photocatalysis in the presence of TiO2 and Fe2O3 were the fastest at 254 nm UV irradiation.

Biodegradation of benzo[a]pyrene in soil by Mucor sp.SF06 and Bacillus sp.SB02 co-immobilized on vermiculite

Two indigenous microorganisms, Bacillus sp. SB02 and Mucor sp. SF06, capable of degrading polycyclic aromatic hydrocarbons （PAHs） were co-immobilized on vermiculite by physical adsorption and used to degrade benzo[a] pyrene （BaP）. The characteristics of BaP degradation by both free and co-immobilized microorganism were then investigated and compared. The removal rate using the immobilized bacterial-fungal mixed consortium was higher than that of the freely mobile mixed consortium. 95.3% of BaP was degraded using the co-immobilized system within 42 d, which was remarkably higher than the removal rate of that by the free strains. The optimal amount of inoculated co-immobilized system for BaP degradation was 2%. The immobilized bacterial-fungal mixed consortium also showed better water stability than the free strains. Kinetics of BaP biodegradation by co-immobilized SF06 and SB02 were also studied. The results demonstrated that BaP degradation could be well described by a zero-order reaction rate equation when the initial BaP concentration was in the range of 10--200 mg/kg. The scanning electronic microscope （SEM） analysis showed that the co-immobilized microstructure was suitable for the growth of SF06 and SB02. The mass transmission process of co-immobilized system in soil is discussed. The results demonstrate the potential for employing the bacterial-fungal mixed consortium, co-immobilized on vermiculite, for in situ bioremediation of BaP.

Antioxidant responses to benzo[a]pyrene,tributyltin and their mixture in the spleen of Sebasticus marmoratus

It has been reported that there is an interaction between Benzo[a]pyrene （BaP）, a widespread carcinogenic polycyclic aromatic hydrocarbon, and tributyltin （TBT）, an organometal used as an antifouling biocide. This study was therefore designed to examine the potential in vivo influence of BaP, TBT and their mixture on splenic antioxidant defense systems of Sebastiscus marmoratus. The fish were exposed to water containing environmentally relevant concentrations of BaP, TBT and their mixture. Spleens were collected for biochemical analysis after exposure for 7, 25, 50 d and after recovery for 7, 20 d. Cotreatment with BaP and TBT for 7 d potentiated the induction of glutathione peroxidase （GPx） activity by BaP or TBT alone. The cotreatment for 25 and 50 d resulted in inhibition of GPx activity, which was similar to the effect of TBT. Splenic glutathione S-transferase （GST） activities were significantly elevated in S. marmoratus exposed to BaP starting from 7 d and remained high up to 25 d. However, no further activity change was found with prolonged exposure. Cotreatment of BaP and TBT primarily inhibited the GST activity, which was similar to the effect of TBT. Cotreatment with BaP and TBT for 25 or 50 d potentiated the depletion of GSH （glutathione） by BaP or TBT alone. MDA （malondialdehyde） contents in spleen of S. marmoratus were not significantly altered compared with the control during the test period. Spleen, as an immune organ, is sensitive to exposure of BaP or TBT. It should have an effective mechanism to counteract oxidative damage. Antioxidative defense systems in spleen of S. marmoratus should be considered as potential biomarkers. Short-term exposure of BaP or TBT could result in induction of antioxidant defense system. A significant decrease of these indices, such as GSH, GST, GPx might indicate more severe contamination.

Isolation of marine benzo[a]pyrene-degrading Ochrobactrum sp. BAP5 and proteins characterization

A bacterial strain BAP5 with a relatively high degradation ability of benzo[a]pyrene （BaP） was isolated from marine sediments of Xiamen Western Sea, China and identified as Ochrobactrum sp. according to 16S rRNA gene sequence as well as Biolog microbial identification system. Strain BAP5 could grow in mineral salt medium with 50 mg/L of BaP and degrade about 20% BaP after 30 d of incubation. Ochrobactrum sp. BAP5 was able to utilize other polycyclic aromatic hydrocarbons （PAHs） （such as phenanthrene, pyrene and fluoranthene） as the sole carbon source and energy source, suggesting its potential application in PAHs bioremediation. The profile of total soluble protein from Ochrobactrum sp. BAP5 was also investigated. Some over- and special-expressed proteins of strain BAP5 when incubated with the presence of BaP were detected by two-dimensional polyacrylamide gel electrophoresis, and found to be related with PAHs metabolism, DNA translation, and energy production based on peptide fingerprint analysis through matrix-assisted laser desorption/ionization-time of flight mass spectrometry.

Phytotoxicity assessment of phenanthrene, pyrene and their mixtures by a soil-based seedling emergence test

Seedling emergence tests were conducted in a meadow brown soil using five plant species(i.e., Chinese cabbage, green onion, tomato, turnip and wheat) to determine the phytotoxicity of phenanthrene, pyrene and their mixtures. The soil was amended with up to 1000 mg/kg soil of phenanthrene or 600 mg/kg soil of pyrene. Seedling emergence and root growth were measured. The results indicated that root elongation was more sensitive than seedling emergence. Root length decreased with increasing phenanthrene or pyrene concentrations(p≤0 05). Phenanthrene was more phytotoxic than pyrene. The lowest observable adverse effect concentration(LOAEC) was 10 mg/kg for phenanthrene when tested with green onion, and 50 mg/kg for pyrene when tested with wheat. Among the five species, wheat was found to be the most sensitive. When amended jointly at or below their respective LOAEC, phenanthrene and pyrene produced a synergistic toxic effect.

Uptake and accumulation of phenanthrene and pyrene in spiked soils by Ryegrass(Lolium perenne L.)

Phytoremediation has long been recognized as a cost-effective method for the removal of polycyclic aromatic hydrocarbons （PAHs） from soil. A study was conducted to investigate the uptake and accumulation of PAHs in root and shoot of Lolium perenne L. Pot experiments were conducted with series of concentrations of 3.31-378.37 mg/kg for phenanthrene and those of 4.22-365.38 mg/kg for pyrene in a greenhouse. The results showed that both ryegrass roots and shoots did take up PAHs from spiked soils, and generally increased with increasing concentrations of PAH in soil. Bioconcentration factors（BCFs） of phenanthrene by shoots and roots were 0.24- 4.25 and 0.17-2.12 for the same treatment. BCFs of pyrene by shoots were 0.20-1.5, except for 4.06 in 4.32 mg/kg treatment, much lower than BCFs of pyrene by roots （0.58-2.28）. BCFs of phenanthrene and pyrene tended to decrease with increasing concentrations of phenanthrene and pyrene in soil. Direct uptake and accumulation of these compounds by Lolium perenne L. was very low compared with the other loss pathways, which meant that plant-promoted microbial biodegradation might be the main contribution to plant-enhanced removal of phenanthrene and pyrene in soil. However, the presence of Lolium perenne L. significantly enhanced the removal of phenanthrene and pyrene in spiked soil. At the end of 60 d experiment, the extractable concentrations of phenanthrene and pyrene were lower in planted soil than in non-planted soil, about 83.24%-91.98% of phenanthrene and 68.53%-84.10% of pyrene were removed from soils, respectively. The results indicated that the removal of PAHs in contaminated soils was a feasible approach by using Lolium perenne L.

In vitro Study on Role of Hsp70 Expression in DNA Damage of Human Embryonic Lung Cells Exposed to Benzo[a]pyrene

Objective Benzo[a]pyrene (B[a]P), a ubiquitous environmental pollutant, is a potent procarcinogen and mutagen that can elicit tumors, leading to malignancy. Heat shock proteins (Hsp) have been shown to protect cells against damages caused by various stresses including exposure to numerous chemicals. Whether Hsps, or more specifically Hsp70, are involved in repair of B[a]P-induced DNA damage is currently unknown. Methods We assessed the potential role of the inducible form of Hsp70 in B[a]P-induced DNA damage of human embryonic lung (HEL) cells using immunoblot and the comet assay (i.e., the single cell gel electrophoresis assay). Results Exposure to B[a]P induced a dose-dependent decrease in the level of Hsp70, but a dose-dependent +-increase in DNA damage both in untreated (control) HEL cells and in cells preconditioned by a heat treatment. Heat preconditioning prior to B[a]P exposure potentiated the effect of B[a]P at a low dose (10 μmol/L), but appeared to be protective at higher doses. There was a negative correlation between Hsp70 level and DNA damage in the non-preheated as well as in the preconditioned cells. Conclusion These data suggest that exposure of HEL cells to B[a]P may induce a dose-dependent reduction in the levels of the inducible Hsp70. The detailed mechanisms for the reduction of Hsp70 levels by B[a]P and the role of Hsp70 in DNA damage under different concentrations of B[a]P remains to be determined.

1-Pyrenecarboxaldehyde thiosemicarbazone:A novel fluorescent molecular sensor towards mercury(Ⅱ) ion

A novel and simple fluorescent molecular sensor,1-pyrenecarboxaldehyde thiosemicarbazone（Hpytsc）,was synthesized.Its higher sensitivity and selectivity to mercury（Ⅱ） ion were studied through absorption and emission channels.The UV-vis spectra show that the increasing mercury（Ⅱ） ion concentrations result in the decreasing absorption intensity.The fluorescence monomer emission of Hpytsc is enhanced upon binding mercury（Ⅱ） ion,which should be due to the 1：1 complex formation between Hpytsc and metal ion.

n-Hexadecane and pyrene biodegradation and metabolization by Rhodococcus sp. T1 isolated from oil contaminated soil

The high-molecular weight polycyclic aromatic hydrocarbons(PAHs) pyrene and typical long chain alkane nhexadecane are both difficult to degrade. In this study, n-hexadecane and pyrene degrading strain Rhodococcus sp. T1 was isolated from oil contaminated soil. Strain T1 could remove 90.81% n-hexadecane(2 vol%) and 42.79% pyrene(200 mg·L^(-1)) as a single carbon within 5 days, respectively. Comparatively, the degradation of pyrene increased to 60.63%, but the degradation of n-hexadecane decreased to 87.55% when these compounds were mixed. Additionally, identification and analysis of degradation metabolites of Rhodococcus sp. T1 in the above experiments showed that there were significant changes in alanine, methylamine, citric acid and heptadecanoic acid between sole and dual substrate degradation. The optimal conditions for degradation were then determined based on analysis of the pH, salinity, additional nutrient sources and liquid surface activity.Under the optimal conditions of pH 7.0, 35 °C, 0.5% NaCl, 5 mg·L^(-1) of yeast extract and 90 mg·L^(-1) of surfactant,the degradation increased in single or dual carbon sources. To our knowledge, this is the first study to discuss metabolite changes in Rhodococcus sp. T1 using sole substrate and dual substrate to enhance the long-chain alkanes and PAHs degradation potential.

Binding of pyrene to dissolved organic matters: fractionation and characterization

Based on a modified Leenheer DOM fractionation scheme, fractionation of DOM from the paddy soil was conducted by using XAD-8 resin into hydrophobic bases(HOB), hydrophobic acids(HOA), acid-insoluble matter(AIM), hydrophobic neutrals(HON) and hydrophilic matter(HIM). In total carbon content of DOM, 35.32% were the HIM and only 0.73% the HOB. However, HOA and AIM altogether occupied 53.45%, while the HON fraction represented 10%. The sorption experiments were conducted to determine the sorption capacity of pyrene on unfractionated DOM and its fractions. Elemental analysis, 1H-NMR and FTIR spectra were carried out on unfractionated DOM and its fractions to examine the relationship between the structure of DOM and partition coefficients(K oc). The results showed that HON had a greater affinity for binding pyrene than other fractions. While HON was characterized by large long-chain alkylate (aliphatic structure). AIM exhibited relative higher K oc values than HOA and HIM, due to much aromatic structure in AIM, while the high content of carboxylic groups of HOA and HIM depressed their binding capacity. This study demonstrated HON is a key subcomponents of DOM in binding of pyrene, in other words, aliphalic structure in DOM play an important role in binding of pyrene.

Significant Positive Correlation of Plasma BPDE-Albumin Adducts to Urinary 1-Hydroxypyrene in Coke Oven Workers

Objective To investigate the application of BPDE-albumin adducts as monitoring biomarkers for coke oven workers exposed to polycyclic aromatic hydrocarbons （PAHs） and to explore possible relationship between BPDE-albumin adducts and urinary 1-hydroxypyrene （1-OHP） levels in them. Methods Thirty-seven coke oven workers from a coke plant and 47 controls without the occupational exposure to PAHs were recruited in this study. The levels of plasma BPDE-albumin adducts and urinary 1-OHP were analyzed using high performance liquid chromatography. Results The median levels of BPDE-albumin adducts （42.10 fmol/mg albumin） and urinary 1-OHP （5.46 μmol/mol creatinine） were significantly higher in coke oven workers than in controls （14.16 fmol/mg albumin, 2.96μmol/mol creatinine, respectively; P〈0.01）. Multiple logistic regression analysis showed that coke oven workers were at higher risk of having BPDE-albumin adduct levels above 25.30 μmol/mg albumin （OR=1.79, P〈0.01） and urinary 1-OHP levels above 4.13 μmol/mol creatinine （OR=2.45, P〈0.05）. There was a positive correlation between the levels of BPDE-albumin adducts and urinary 1-OHP in all subjects （rs=0.349, P〈0.01）. Conclusion BPDE-albumin adduct is a useful biomarker for monitoring long-term exposure to PAHs, and plasma BPDE-albumin adducts level is significantly correlated to urinary 1-OHP levels in coke oven workers.

Combined effect of tributyltin and benzo[a] pyrene on the levels of sex hormone and vitellogenin in female Sebastiscus marmoratu

Tributyltin（TBT）, an organometal used as an antifouling biocide, has been reported to induce masculinization of fish. Benzo [a]pyrene （BaP）, a widespread carcinogenic polycyclic aromatic hydrocarbon, has been reported that its microsomal metabolites can produce an estrogenic response when tested in vitro. This study was therefore designed to examine the potential in vivo influence of TBT, BaP and their mixture on sex hormone levels in serum of Sebastiscus marmoratus, which were given 2 separate intraperitoneally （ip） injections（a single injection every 7 d） of TBT（0.5, 1, 5 and 10 mg/kg）, BaP（0.5, 1, 5 and 10 mg/kg）, or both in combination（0.5, 1, 5 and 10 mg/kg）; control fish received olive oil vehicle only. Six days after the 2nd injection, serum samples were collected and analyzed for sex hormone levels and alkali labile protein phosphorus （ALPP）, which is related to the yolk precursor protein vitellogenin. The pollutants at all doses significantly reduced serum testosterone, estradiol and ALPP content after 2 injections compared with the corresponding controls. The reduction of the estradiol levels should be response for the decrease of the vitellogenin levels. The results in the present study suggested that aromatase seems not the major target acted by TBT and BaP in fish. This study demonstrated that TBT or BaP exposure both inhibit the reproductive potential in female Sebastiscus marmoratus. Combined effect of TBT and BaP on the serum testosterone, estradiol and ALPP was not antagonism from the anticipation.