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.

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.

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.

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.

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.

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.

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.

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.

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.

Effects of benzo(a)pyrene exposure on the antioxidant enzyme activity of scallop Chlamys farreri

Scallop Chlamys farreri was exposed to different concentrations of benzo(a)pyrene (BaP) (0.5 μg/L, 1.0 μg/L, 10.0 μg/L and 50.0 μg/L) for 30 days in seawater. The 7-ethoxyresorufin O-deethylase (EROD) activity was significantly induced, and increased with the increasing BaP concentration. The glutathione-S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), Glutathione peroxidase (GPx) activities increased in short time at low concentration of BaP, and was significantly depressed at high concentrations. Scallop gill was more sensitive to BaP than the digestive gland, and the digestive gland was the main tissue to deal with oxyradicals. The contents of malondialdehyde (MDA) increased with the exposure time and there was a positive correlation (concentration-effect) between the MDA content and the concentration of BaP. The biomarkers determined in this experiment had important roles in detoxification, and showed great potential as biomarkers for oxidative stress. Controlled laboratory experiments designed to simulate field exposure scenarios are particularly useful in ascertaining biomarkers suitable for use with complex contaminant mixtures in the marine environment.

Effect of Benzo[a]pyrene on Detoxification and the Activity of Antioxidant Enzymes of Marine Microalgae

The objective of this study was to examine the effect ofbenzo[a]pyrene （BaP） on the detoxification and antioxidant systems of two microalgae, Isochrysis zhanjiangensis and Platymonas subcordiformis. In our study, these two algae were exposed to BaP for 4 days at three different concentrations including 0.5 μg L-1 （low）, 3 μg L-1 （mid） and 18 μg L-1 （high）. The activity of detoxi- fication enzymes, ethoxyresorufin O-deethylase （EROD） and glutathione S-transferase （GST） increased in P subcordiformis in all BaP-treated groups. In 1. zhanjiangensis, the activity of these two enzymes increased at the beginning of exposure, and then de- creased in the groups treated with mid- and high BaP. The activity of antioxidant enzyme superoxide dismutase （SOD） increased in/. zhanjiangensis in all BaP-treated groups, and then decreased in high BaP-treated group, while no significant change was observed in P subcordiformis. The activity of antioxidant enzyme catalase （CAT） increased in I. zhanjiangensis and P subcordiformis in all BaP- treated groups. The content of malondialdehyde （MDA） in Isochrysis zhanjiangensis increased first, and then decreased in high BaP-treated group, while no change occurred in P. subcordiformis. These results demonstrated that BaP significantly influenced the activity of detoxifying and antioxidant enzymes in microalgae. The metabolic related enzymes （EROD, GST and CAT） may serve as sensitive biomarkers of measuring the contamination level of BaP in marine water.

Simple Fluorimetric Determination of Benzo[a]pyrene in Cigarette Smoke without Preseparation Procedure

Constant-energy synchronous fluorimetry was used for the identification of benzo[a]pyrene in mixtures with a detection limit of 1.34 nmol/L. The recovery experiments in cigarette smoke samples have also obtained satisfactory results of 99.1-103.5% for benzo[a]pyrene.

Different Patterns of Cyclin D1/CDK4-E2F-1/4 Pathways in Human Embryo Lung Fibroblasts Treated by Benzo[a]pyrene at Different Doses

Objective To investigate the roles of the cyclin D1/CDK4 and E2F-1/4 pathways and compare their work patterns in cell cycle changes induced by different doses of B[a]E Methods Human embryo lung fibroblasts （HELFs） were treated with 2 μmol/L or 100 μmol/L B[a]P which were provided with some characteristics of transformed cells （T-HELFs）. Cyclin D l, CDK4 and E2F-1/4 expressions were determined by Western blotting. Flow cytometry was used to detect the distribution of cell cycle. Results After B[a]P treatment, the proportion of the first gap （G 1） phase cells decreased. CDK4 and E2F-4 expression did not change significantly. In 2 μmol/L treated cells, a marked overexpression of cyclin D1 and E2F-1 was observed. However, in T-HELFs overexpression was limited to cyclin D1 only, and no overexpression of E2F-1 was observed. The decreases of G1 phase in response to B[a]P treatment were blocked in antisense cyclin D1 and antisense CDK4 transfected HELFs （A-D1 and A-K4） and T-HELFs （T-A-D1 and T-A-K4）. After 2 μmol/L B[a]P treatment, overexpression of E2F-1 was attenuated in A-D1, and E2F-4 expression was decreased significantly in A-K4. In T-A-D1 and T-A-K4, E2F-4 expression was increased significantly, compared with T-HELFs. The E2F-1 expression remained unchanged in T-A-D1 and T-A-K4. Condusions Cyclin DI/CDK4-E2F-1/4 pathways work in different patterns in response to low dose and high dose B[a]P treatment. In HELFs treated with 2 μmol/L B[a]P, cyclin D1 positively regulates the E2F-1 expression while CDK4 negatively regulates the E2F-4 expression; however, in HELFs treated with 100 μmol/L B[a]P, both cyclin D1 and CDK4 negatively regulate the E2F-4 expression.

Modulation of Behavior and Glutamate Receptor mRNA Expression in Rats after Sub-chronic Administration of Benzo(a)pyrene

Objective The present study aimed to test whether exposure to benzo（a）pyrene [B（a）P] affects spatial learning and short-term memory by modulating the expression of the Gria1 and Grin2a glutamate receptor subunit genes in the hippocampus.Methods Thirty-six 21-24-day-old,male rats were randomly assigned into high-,medium-,and low-dose toxin exposure groups （6.25,2.5,and 1 mg/kg,respectively） and a control group,each containing nine rats.The behavioral performance of adult rats exposed to sub-chronic administration of B（a）P was monitored by learning and memory tests （Morris water maze）.Real-time PCR assays were used to quantify Gria1 and Grin2a gene expression in the hippocampus.Results At medium and high doses,B（a）P impaired spatial learning performance.The crossing-platform-location frequency and the time spent swimming in the platform area,which both relate to short-term memory,were significantly decreased in B（a）P-treated rats compared with controls.The level of Gria1 mRNA increased 2.6-5.9-fold,and the level of Grin2a mRNA increased 10-14.5-fold,with a greater fold increase associated with higher doses of B（a）P.Conclusion We demonstrated that sub-chronic administration of B（a）P inhibits spatial learning and short-term memory,and increases Gria1 and Grin2a expression in the hippocampus.This suggests a relationship of B（a）P exposure levels with Gria1 and Grin2a expression and impairment of short-term and spatial memory.

Effects of Benzo(a)pyrene on the Contractile Function of the Thoracic Aorta of Sprague-dawley Rats

Abstract Objective To evaluate the possible vascular effects of an environment carcinogen benzo（a）pyrene （BaP）. Methods The cytotoxicit of BaP and rat liver S9 （0.25 mg/mL）-activated BaP were examined by MTT assay. Thoracic aortic rings were dissected from Sprague-Dawley rats. Contraction of aortic rings was induced by 60 mmol/L KCl or 10-6 mol/L phenylephrine （PE） in an ex-vivo perfusion system after BaP （100 tlmol/L） incubation for 6 h. [Ca^2＋]i was measured using Fluo-4/AM. For in-vivo treatment, rats were injected with BaP for 4 weeks （10 mg/kg, weekly, i.p.）. Results BaP （1-500 μm） did not significantly affect cell viability; S9-activated BaP stimulated cell proliferation. BaP did not affect the contractile function of endothelium-intact or -denuded aortic rings. BaP did not affect ATP-induced （[Ca2＋]i） increases in human umbilical vein endothelial cells. In BaP-treated rats, heart rate and the number of circulating inflammatory cells were not affected. Body weight decreased while blood pressure increased significantly. The maximum aortic contractile responses to PE and KCI and the maximum aortic relaxation response to acetylcholine were significantly decreased by 25.0%, 34.2%, and 10.4%, respectively. Conclusion These results suggest, in accordance with its DNA-damaging properties, that metabolic activation is a prerequisite for BaP-induced cardiovascular toxicity.

EROD activities of liver in Mugil so-iuy exposed to benzo(a)pyrene,pyrene and their mixture

The effects on hepatic EROD (7-ethoxyresorufin O-deethylase) in Mugil so-iuy exposedto benzo(a)pyrene (BaP), pyrene and their mixtures of equal concentration were investigated, at concentrations of 0.1, 1.0, 5.0, 10.0, 50.0μg/dm3, in experimental condition. Time-effects and dose-response of the biochemical indexs were observed. The results showed that the hepatic EROD activities were induced by the exposure of BaP, pyrene and their mixtures at high concentration. Dose-response connections were that the hepatic EROD activities were elevated with increasing concentration of the pollutants. The combined effect of BaP and pyrene at 1:1 concentration ratio on hepatic EROD activity was antagonism.

Immunotoxicity Effect of Benzo[α]Pyrene on Scallop Chlamys farreri

The toxic effects of benzo[α]pyrene （B[α]P） at different concentrations （0.1, 0.5, 1, 2.5 and 7.5 μgL^-1） on scallop （Chlamys farreri） immune system were studied. The results showed that B[α]P had significant toxic effects on the haemocyte counts, neutral red uptake, phagocytosis, bacteriolytic and antibacterial activity （P〈0.05）, while the seawater control and acetone control had no significant differences. The haemocyte counts, neutral red uptake, phagocytosis and bacteriolytic activity in all B[α]P treatment groups as well as antibacterial activity in groups of 0.5, 1, 2.5 and 7.5 μgL^-1·B[α]P decreased significantly （P〈0.05）. Some of these indices tended to be stable on the sixth day and others on the ninth day, and the indices showed clear time- and concentration-response to B[α]E Bactenolytic activity in 0.1 μgL^-1 B[α]P treatment group and antibacterial activity in 0.1 ggLl and 0.5 μgL^-1 B[α]P treatment groups increased at the beginning of exposure and reached their peaks on day 1 and day 6, respectively. Following that, both activities decreased gradually and became stable after day 9. When all the indices reached stability, they were significantly lower than those in control group （P〈0.05）, except for antibacterial activity in 0.1 μgLl B[α]P treatment group （P〉0.05）. Thus, B[α]P has evident toxic effects on scallop immune system, which supports the view that a relationship exists between pollution and lmmunomodulation in aquatic organisms.

Preliminary Characterizations of a Carbohydrate from the Concentrated Culture Filtrate from <i>Fusarium solani</i>and Its Role in Benzo[a]Pyrene Solubilization

In order to investigate the mechanism of benzo[a]pyrene uptake by a filamentous fungus Fusarium solani, a biochemical characterization of its concentrated culture filtrate has been conducted. The preparation contained approximately (w/w): 50% of total carbohydrate, 6.5% of uronic acid and 6% protein, as determined by colorimetric tests. Gel filtration and anion-exchange chromatographic profiles indicated that the main product of the culture filtrate was a glycoprotein, which contained mannose, glucose and galactose in an approximate molar ratio of 1.5: 0.8: 1. The polysaccharide fraction of the culture filtrate was prepared by treatment with proteinase K, followed by gel-filtration chromatography. Its chemical structure was studied by methylation analysis, gas-liquid chromatography-mass spectrometry (GC-MS) and Nuclear Magnetic Resonance spectroscopy (NMR). The major carbohydrate was a polymer of β-(1 → 6)-linked galactofuranose units fully branched at positions O-2 by single residues of α-glucopyranose. The Fusarium concentrated culture filtrate increased 4-fold the BaP solubilization in comparison with its aqueous solubility and suggested that the carbohydrate present in this filtrate should probably be involved in this enhancement. Our findings point out the potential role of fungal glycoproteins in PAH microbial bioavaibility, an important step for PAH biodegradation.

Inhibitory Effect of Green Tea Extract on the Carcinogenesis Induced by Asbestos Plus Benzo(a)pyrene in Rat

In this experiment lung carcinoma was induced by crocidolite plus benzo(a)pyrene in rat. From the cancer models, we observcd that the incidcnce (16.0%) of lung carcinomas was lowel, and the survival time (376 days) of the first case of carcinoma and the mean survival time (758 days) of the rats with carcinoma were higher in the group of rats drinking 2% green tea extract for life than in the positive group (without drinking green tea extract). The mortality ratio (0.5047) was smaller in the cxperimental group than in the positive control group, and the survival curve of the experimental group significantly raised up, in comparison with the positive group.