• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.11 no.2
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• pp.118-125
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• 2001

Recently, biochemical analysis using hemoglobin adduct is frequently performed to evaluate the exposure to chemical carcinogens. However, data on the effect of co-exposure with other chemicals on hemoglobin adduct formation are seldom provided. The objective of this study is to evaluate the effects of pretreatment of ethanol(EtOH) and phenobarbital(PB), which are known to affect metabolism of xenobiotics, on the formation of hemoglobin adducts in the rats(Sprague-Dawley) administered benzidine(BZ). The experimental rats were divided into control, EtOH, and P8 groups. Rats were pretreated with EtOH or PB 24 hours before the oral administration of BZ. Blood sampling was taken before the administration of the chemicals and 0.5, 3, 6, 9, 12, 24, 48, 72, 96, and 144 hours after the administration of the BZ in 5 rats each. The blood was separated into hemoglobin and plasma immediately after taking the blood samples, and the adducts were undergone basic hydrolysis to convert them into aromatic amines. Hydrolyzed BZ, monoacetylbenzidine (MABZ), and 4-aminobiphenyl(4ABP) were separated by reversed-phase liquid chromatography without derivatization, and quantitative analyses of them were performed by a highperformance liquid chromatograph equipped with electrochemical detector. The quantitative amount of the metabolites was expressed by hemoglobin binding index(HBI), BZ-, MABZ-, and 4ABP-HBI of EtOH and PB groups were increased more than those of control group. These results are attributable to the fact that EtOH and PB induced N-hydroxylation related to the hemoglobin adduct formation. The ratio of N-acetylation (viz, MABZ-HBI/BZ-HBI) showed no significant difference between EtOH group and control group. It means that EtOH increased N-hydroxylation and N-acetylation in a similar degree. The N-acetylation ratio of PB group was relatively lower than control group because the PB increased N-hydroxylation induction. The N-acetylation ratios of all groups were higher than 1 during the entire experimental period. This result suggests that the effects of EtOH or PB need to be considered in the biochemical monitoring for the assessment of intermittent exposure of benzidine.

• Journal of the Korean Society of Tobacco Science
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• v.32 no.1
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• pp.1-11
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• 2010

This study was carried out to investigate the influence of stem materials such as black stem on the quality of expended stem and cigarettes. Normal and black stem were separated by tobacco scan and then, those stems were expanded after treating with their respective stem casings. Total sugar, ether extract, ash contents and pH were slightly low in black stem compared with normal stem. However, the number of bacteria and fungi ratio were remarkably higher in black stem than that of normal stem. As compared with normal stems, ratio of rushed stem in rolled process was approximately 2 times higher in black stem with the consequency that the filling capacity of black stem was decreased. The ratio of large particles (> 3.35 mm) of expanded black stem showed decreasing tendency and small particles rate (1.40 mm <) was increased compared with normal stem. When expanded stems were prepared using stem containing 5 levels (0, 10, 20, 30 and 100 %) of black stem, the filling capacity was decreased and static burning rate was significantly decreased with increasing expanded black stem rate. However, the weight and hardness of cigarettes were slightly increased with increasing expanded black stem rate. The contents of phenol compounds, aromatic amines and carbonyl compounds in the cigarette mainstream smoke from the cigarette which was manufactured with various ratio of expended black stem, were gradually increased with increasing expanded black stem rates. Also, the cytotoxicity and the mutagenicity of the TPM were significantly increased with increasing expanded black stem rate. The sensory test result showed that cigarettes blended with 10 and 30 % level of black stem rate was exhibited significantly high sensory attributions such as off-taste, impact, hotness, bitterness and irritation as compared with cigarette blended with normal stem, while smoke fullness and cleanness were slightly decreased with increasing expanded black stem rates. The number of brown spots on cigarettes paper was 2 to 3 times high in cigarettes containing black stem than that of cigarette made from normal stem and were high with increasing black stem rate. The overall assessment in this study suggest, that black stem should not be used because of bad quality of expanded stem and high toxicological activity of cigarette mainstream smoke.

• Journal of Preventive Medicine and Public Health
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• v.35 no.3
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• pp.229-235
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• 2002

Objectives : To evaluate the effects on the formation of benzidine-hemoglobin, and benzidine metabolite-hemoglobin adducts, caused by pretreatment with the known xenobiotic metabolism effectors, ethanol and phenobarbital, in rats administered Direct Black 38 dye. Methods : The experimental rats were divided into three groups: a control group, an ethanol group and a phenobarbital group. Rats were pretreated with ethanol (1g/kg) or phenobarbital (80mg/kg) 24 hours prior to the oral administration of Direct Black 38 (0.5mmol/kg), with the control group being administered the same amount of distilled water. Blood samples were obtained from the vena cava of 5 rats from each group prior to, and at 30 min, 3h, 5h, 9h, 12h, 24h, 48h, 72h, 96h, and 144h following the oral administration of Direct Black 38. Directly after sampling the blood was separated into hemoglobin and plasma, with the adducts being converted into aromatic amines by basic hydrolysis. Hydrolyzed benzidiene, monoacetylbenzidine and 4-aminobiphenyl were analyzed by reverse-phase liquid chromatography with an electrochemical detector, The quantitative amount of the metabolites was expressed by the hemoglobin binding index (HBI). Results : In the ethanol group, benzidine-, monoacetylbenzidine-, and 4-aminobiphenyl-HBI were increased to a greater extent than those in the control group. These results were attributed to the ethanol inducing N-hydrgxylation, which is related to the formation of the hemoglobin adduct, In the phenobarbital group, all the HBIs, with the exception of the benzidine-HBI, were increased to a greater extent than those of the control group. These results were attributed to the phenobarbital inducing N-hydroxylation related to the formation of the hemoglobin adduct. The N-acetylation ratio was only increased with the phenobarbital pretreatment due to the lower benzidine-HBI of the phenobarbital group compared to these of the control and ethanol groups. The N-acetylation ratios for all groups were higher than f for the duration of the experimental period. Although the azo reduction was unaffected by the ethanol, it was inhibited by the phenobarbital, The ratio of the benzidine-HBI in the phenobarbital group was lower than those of the ethanol the control groups for the entire experiment. Conclusion : Our results indicate that both ethanol and phenobarbital increase the formation of adducts by the induction of N-hydroxylation, but also induced N-acetylation. Phenobarbital decreased the formation of benzidine-HBI due to the decrease of the azo reduction. These results suggest that the effects or ethanol and phenobarbital need to be considered in the biochemical monitoring of Direct Black 38.

• The Korean Journal of Pharmacology
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• v.31 no.3
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• pp.299-311
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• 1995

Platelets resemble monoaminergic neurons in several respects, i.e. the uptake of 5-HT and its inhibition, the subcellular storage and release of 5-HT, and the metabolism of aromatic amines brought about by monoamine oxidase. And the 5-HT content of rabbit platelets is well known to be about 40 times higher than that of human platelets. Therefore, this study was carried out to investigate the influences of amitriptyline (AMT) and sertraline (SRT) on the aggregation, contents of signaling second messengers, and protein phosphorylations of rabbit platelets in response to thrombin, 0.25 unit/ml, comparing with those of chlorpromazine (CPZ). Thrombin-induced aggregation was inhibited by SRT $(IC50:4.37{\times}10^{-5}\;M)$, CPZ $(IC50:5.76{\times}10^{-5}\;M)$, and AMT $(IC50:1.15{\times}10^{-4}\;M)$, respectively, and the aggregation by A23187 $(1.0\;{\mu}M)$ or PMA (320 nM) was also inhibited by SRT, CPZ, and AMT. AMT, SRT, and CPZ had little affects on basal contents of platelet $TXB_2$ and $PGE_2$, but all of them inhibited the thrombin-induced increase of $TXB_2$. Thrombin did not change the platelet contents of cAMP and cGMP. CPZ, AMT, and SRT produced the slight decrease of basal cAMP content, and their effects were not affected by thrombin-treatment. But SRT and AMT moderately increased the basal cGMP content, and the cGMP content of thrombin-stimulated platelets was gradually increased by the pretreatment with SRT, AMT, and CPZ. Particularly, the SRT-dependent increase of the cGMP content was notable. Platelet $Ins(1,4,5)P_3$ content was rapidly increased up to a plateau within 10 sec after thrombin-stimulation, AMT, SRT, and CPZ increased the basal $Ins(1,4,5)P_3$ content, and the thrombin-dependent increase was enhanced by pretreatment with CPZ and AMT, but was blunted by SRT. Platelet $[Ca^{2+}]_i$, was rapidly increased up to a peak level within 20 sec after thrombin-stimulation. The increase of $[Ca^{2+}]_i$ was sisnificantly inhibited by AMT, SRT, and CPZ. Thrombin- or PMA-induced phosphorylations of platelet $41{\sim}43\;kDa$ and 20 kDa proteins were significantly inhibited by AMT, SRT, and CPZ. These results suggest that the antiplatelet activities of AMT and CPZ may be considerably attributed to the inhibition of protein kinase C activity, and the activity of SRT may be associated with the inhibitory effect on the thrombin-induced increase of $Ins(1,4,5)P_3$ and the increasing effect on the cGMP content of ptatelets. Therefore, it seems to be evident that AMT and SRT may produce their antidepressant activity, at least, partly through the inhibition of protein kinase C activity or the increase of resting $Ins(1,4,5)P_3$, content and in case of SRT, to a lesser extent, via the increase of cGMP in the brain.