2007 Vol. 20, No. 3

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Significant Positive Correlation of Plasma BPDE-Albumin Adducts to Urinary 1-Hydroxypyrene in Coke Oven Workers
HONG WANG, XIAO-BO YANG, AI-LIN LIU, HONG-YAN ZHEN, LIANG GUO, HUA-SHAN LIANG, YONG-YI BI, YUN BAI, YONG-WEN CHEN, TANG-CHUN WU
2007, 20(3): 179-183.
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,2.96 μmol/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 prnol/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.
Production and Characterization of Monoclonal Antibody Against Recombinant Human Erythropoietin
JIE-BO MI, JIN YAN, XIAO-JIE DING, ZHEN-QUAN GUO, MEI-PING ZHAO, WEN-BAO CHANG
2007, 20(3): 184-188.
Objective To produce specific monoclonal antibody(mAb)against recombinant human erythropoietin(rHuEPO)for development of higmy efficient methods for erythropoietin detection in biological fluids.Methods rHuEPO was covalently coupled with bovine serum albumin(BSA)and the conjugate was used to immunize mice to produce specific mAb against rHuEPO based on hybridoma technology.The obtained F3-mAb was characterized by enzyme-linked immunosorbent assay (ELISA),SDS-PAGE and Western blot.Results The isotype of F3-mAb Was found to be IgM with an affinity constant of 2.1x108 L/mol.The competitive ELISA using the obtained IgM showed a broader linear range and lower detection limit compared with previous work.Conclusions The modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO.
Effect of Lidamycin on Telomerase Activity in Human Hepatoma BEL-7402 Cells
RUI-JUAN GAO, YUE-XIN LIANG, DIAN-DONG LI, HONG-YIN ZHANG, YONG-SU ZHEN
2007, 20(3): 189-197.
Objective To investigate the effect of lidamycin(LDM)on telomerase activity in human hepatoma BEL-7402 cells under the condition of LDM inducing mitotic cell death and senescence.Methods Chromatin condensation was detected by co-staining with Hoechst 33342 and PI.Cell multinucleation was observed by Giemsa staining and genomic DNA was separated by agarose gel electrophoresis.Fluorescent intensity of Rho123 Was determined for mitochondrial membrane potential.MTT assay and SA-β-gal staining were employed to analyze the senescence-like phenotype.The expression of proteins was analyzed by Western blot.Telomerase activity was assayed by telomerase PCR-ELISA.Results Mitotic cell death occurred in LDM-treated cells characterized by unique and atypical chromatin condensation,multinucleation and increased mitochondrial membrane potential.However,no apoptotic bodies or DNA ladders were found.In addition,apoptosis-related proteins remained nearly unaltered.Senescence-like phenotype was identified by increased and elongated size of cells,growth retardation,enhanced SA-β-gal activity and the changes of senescence-related protein expression.Telomerase activity markedly decreased (P<0.01)in LDM-treated hepatoma BEL-7402 cells. Conelusion Mitotic cell death and senescence could be triggered simultaneously or sequentially after exposure of hepatoma BEL-7402 cells to LDM.The decrease in telomerase activity may play a key role in the defective mitosis and aging morphology.Further investigation of detailed mechanism is needed.
Treatment of Scrapie Pathogen 263K With Tetracycline Partially Abolishes Protease-resistant Activity in vitro and Reduces Infectivity in vivo
YAN-JUN GUO, JUN HAN, HAI-LAN YAO, BAO-YUN ZHANG, JIAN-MEI GAO, JIN ZHANG, XIN-LI XIAO, XIAO-FAN WANG, WEI-QIN ZHAO, DE-XIN WANG, XIAO-PING DONG
2007, 20(3): 198-202.
Objective To study the possible effect of tetracycline on protease-resistant activity in vitro and infectivity in vivo of a scrapie strain 263K.Methods Scrapie pathogens were incubated with tetracycline at different concentrations for various periods of time and protease-resistant PrP signals were evaluated with proteinase K-treatment and Western blots.The preparations treated with tetracycline were intracerebrally inoculated into golden hamsters and typical TSE manifestations were noted.PrPSc in brain tissues of the infected animals was detected by PrP specific Western blot assays. Results Protease-resistant PrP was significantly reduced in or removed from the preparations treated with tetracycline in a dose-dependant manner.Compared with the control group after incubated for 53.75±0.50 days,the preparations treated with 5 mmol/L and 20 mmol/L tetracycline prolonged the incubation time of 61.5±1.73 and 59.5±0.58 days (P<0.05). Conclusion Treatment of scrapie pathogen 263K with tetracycline reduces or removes its protease-resistant activity in vitro.
Trace Metal Levels in Lichen Samples From Roadsides in East Black Sea Region, Turkey
OZGUR DOGAN ULUOZLU, KADIR KINALIOGLU, MUSTAFA TUZEN, MUSTAFA SOYLAK
2007, 20(3): 203-207.
Objective To determine the metal contents of lichen species from East Black Sea region of Turkey for investigation of trace metal pollution sourced traffic.Methods The levels of copper,cadmium,lead,zinc,manganese,iron,chromium,nickel,cobalt,palladium in lichen samples collected from East Black Sea region of Turkey were determined by flame and graphite furnace atomic absorption spectrometry after microwave digestion method.The accuracy of the method was corrected by standard reference material(NIST SRM IAEA-336 Lichen). Results The contents of investigated trace metals in lichen samples were 7.19-22.4 μg/g for copper,0.10-0.64 μg/g for cadmium,4.03-44.6 μg/g for lead,14.5-41.8 1.μg/g for zinc,25.8-208 μg/g for manganese,331-436 μg/g for iron,1.20-3.01 μg/g for chromium,1.48-3.90 μg/g for nickel,0.20-3.55 μg/g for cobalt.0.11-0.64 μg/g for palladium.The results were compared with the literature values.Conclusion Some lichen species such as Xanthoparmelia conspersa,Xanthoria calcicola,Peltigera membranacea,and Physcia adscendens are accumulated trace metals at a high ratio.