Inter-individual variation of smoking-related DNA adducts in lymphocytes-relationship to mRNA levels for CYP1A1 and DNA repair enzymes.

The measurement of DNA adducts is a useful indicator for environmental carcinogen exposure monitoring. To clarify the effect of metabolic activation and DNA repair system on the inter-individual variation of DNA adduct levels, aromatic DNA adducts and mRNA expression of metabolic and repair enzymes were measured in 43 human lymphocytes. Aromatic DNA adducts were measured by the nuclease P1 postlabelling method. The metabolic activation enzyme; cytochrome P4501A1 (CYPIA1), and the repair enzyme; excision repair cross complimenting gene (ERCC1), and the xeroderma pigmentosum C group cell gene (XPCC), mRNA expression were measured by the reverse transcription-PCR method. The mean adduct levels were 1.01 ± 0.49 in 43 subjects. There was a positive correlation between DNA adducts and CYP1A1 mRNA (r = 0.33, p = 0.12). DNA adduct levels had a positive correlation with ERCC1 (r = 0.35, p = 0.03) and a negative correlation with XPCC mRNA levels (r = -0.28, p = 0.07). We found Brinkman index, CYP1A1 genotypes, CYP1A1 mRNA and XPCC mRNA as a predictor for log DNA adduct levels in multivariate analysis. Metabolic activation and the repair system may explain the inter-individual variation of DNA adducts in lymphocytes.

Effects of genetic polymorphism of metabolic enzymes, nutrition, and lifestyle factors on DNA adduct formation in lymphocytes.

Although cigarette smoking is one major determinant of lung carcinogenesis, not all smokers develop cancer. This phenomenon is due to individual variation in genetic susceptibility to carcinogens, nutrition, and lifestyle. Previous studies have shown that genetic polymorphism of metabolic enzymes and plasma micronutrients are associated with lung cancer risk. DNA adducts may serve as a molecular dosimeter for exposure to carcinogens. In this cross-sectional study, we analyzed the blood samples of 158 subjects to evaluate the effects of polymorphisms of cytochrome P450 1A1 (CYP1A1), glutathione S-transferase M1 (GSTM1), T (GSTT), N-acetytransferase 2 (NAT2), and aldehyde dehydrogenase 2 (ALDH2) as well as the effects of plasma beta-carotene and alpha-tocopherol on lymphocyte DNA adducts measured by 32P-postlabeling analysis. The DNA adduct level of smokers (mean +/- SD, 1.26 +/- 0.79/10(8) nucleotides) was significantly higher than that of nonsmokers (0.87 +/- 0.33, P = 0.007). Smokers with CYP1A1 minor homozygotes and GSTM1 null genotypes had a significantly higher level of DNA adducts than those without (P = 0.027 for homozygotes, P = 0.049 for heterozygotes). Smokers with NAT2 minor homozygotes also tended to have a higher DNA adduct level than those with heterozygotes and wild alleles, but the difference was not statistically significant. The DNA adduct level of smokers with ALDH2 heterozygotes was significantly higher than that of smokers with minor homozygotes (P = 0.045). When smokers were divided into "high" and "low" groups according to mean level of plasma beta-carotene or alpha-tocopherol, in the low beta-carotene group, the subjects with CYP1A1 minor homozygotes had higher DNA adduct levels than those with other CYP1A1 genotypes. Smokers with GSTT null genotype and high beta-carotene tended to have a higher DNA adduct level than those with GSTT present and high beta-carotene (P = 0.07), and those with GSTT null genotype and low beta-carotene (P = 0.07). There was weak correlation between DNA adduct level and number of cigarettes smoked per day in the low plasma beta-carotene group (r = 0.28, n = 36, p < 0.1). These results suggested that polymorphisms of CYP1A1, GSTM1, T, NAT2, and ALDH2, and plasma beta-carotene may modulate the level of DNA adducts.

Neurotropic melanoma invading the median nerve.

We report a case of acral lentiginous melanoma of the right thumbnail bed which demonstrated characteristic intraneural invasion and extension along the median nerve. Four years after amputation of the involved thumb, a melanotic tumor recurred on the right thenar. Radiation therapy was given. The tumor invaded the median nerve, however, causing progressive pain and paralysis of the right hand. Despite right arm amputation, the tumor extensively metastasized, and the patient died three years later. Histopathologically, the tumors were characterized by extensive proliferation of spindle-shaped cells forming neuroid fascicles especially prominent in the metastatic region. Tumor cells were positive immunohistochemically with S-100 protein antisera.

Evaluation of organic solvent-induced inflammation modulated by neuropeptides in the abdominal skin of hairless rats.

In this study, the severity and time course of inflammation induced by 4 organic solvents (acetone, cyclohexane, toluene and m-xylene), and the effect of neuropeptides during the inflammation were investigated in the hairless rat abdominal skin. Plasma extravasation used as a parameter of inflammation was measured by Evans blue and 125I-bovine serum albumin (BSA). Total volume of plasma extravasation induced by 4 organic solvents in 240-min exposure was as follows: toluene > m-xylene > cyclohexane > acetone = 0. While hydrophobic solvents (toluene, m-xylene, cyclohexane) induced plasma extravasation, the hydrophilic solvent, acetone, did not induce plasma extravasation. It was suggested that the severity and time course of plasma extravasation depend on chemical characteristics of the organic solvents. In immunohistochemical study, substance P (SP)-immunoreactive nerve fibers (IRNF) and calcitonin gene-related peptide (CGRP)-IRNF were intact during 240-min exposure to acetone. In contrast, cyclohexane, toluene, and m-xylene reduced the number of SP-IRNF and CGRP-IRNF in 10 min exposure and further reduced immunoreactivity. In hairless rats treated with systemic capsaicin, the above plasma extravasation was significantly reduced, along with SP-IRNF and CGRP-IRNF; however, protein gene product 9.5 (PGP 9.5)-IRNF was nearly intact. These results indicated that certain organic solvents induce instance of inflammation that vary widely in terms of their severity and time course, and that these differences are correlated with neuropeptides.

Lymphocytes,DNA adducts and genetic polymorphism for metabolic enzymes in low dose cigarette smokers.

The aim of this study was to investigate the relationship between genetic polymorphism of metabolic enzymes and DNA adduct levels in lymphocytes of low dose cigarette smokers (less than 20 cigarettes per day). We previously reported the effects of cytochrome P4501A1 (CYP1A1) and glutathione S-transferase M1 (GSTM1) on lymphocyte DNA adducts. This time we considered not only CYP1A1 and GSTM1 but also cytochrome P4502E1 (CYP2E1) and glutathione S-transferase T1 (GSTT1). DNA adducts in lymphocytes obtained from low dose cigarette smokers (n = 41) and nonsmokers (n = 56) were measured by the 32P-postlabelling method. The adduct levels were compared regarding smoking status and polymorphic genotypes of these four enzymes. The mean SD of DNA adduct levels in all low dose cigarette smokers and non-smokers was 1 05 0 83 per 108 nucleotidesand 0 85 0 35 per 108 nucleotides, respectively. In low dose cigarette smokers, adduct levels were higher in the rare homozygous (MM) for CYP1A1-exon 7 polymorphism compared with the other types such as common homozygous (WW) and heterozygous (WM). CYP1A1-WM, MM in combination with GSTM1 null showed highest adduct levelamong low smokers. The low smokers with rare homozygous for CYP2E1 Dra1 polymorphism tended to have lower adduct levels than wild types. Low dose cigarette smokers with combined GSTM1 null and T1 null had a higher tendency for adduct levels than others. However none of the differences reached statistical significance.

Relationship between plasma concentrations of beta-carotene and alpha-tocopherol and life-style factors and levels of DNA adducts in lymphocytes.

beta-Carotene and alpha-tocopherol have been thought to reduce risk of lung cancer. Whether beta-carotene and alpha-tocopherol influence human DNA adducts, indicators of biologically effective doses of carcinogens, has been seldom studied. In this cross-sectional study, we measured plasma beta-carotene and alpha-tocopherol in 192 healthy men and DNA adducts in lymphocytes in 104 of the subjects. Because genetic polymorphism of cytochrome P-4501A1 (CYP1A1) and glutathione S-transferase M1 (GSTM1) has been associated with interference in formation of reactive intermediates and detoxification of polycyclic aromatic hydrocarbons, we also obtained data concerning genetic polymorphism of CYP1A1 and GSTM1. In multiple regression analysis, parameters such as alcohol consumed per day, high-density lipoprotein cholesterol, Quetelet index, and cigarettes smoked per day were correlated inversely, whereas age, plasma alpha-tocopherol, and intake frequency of fruits were correlated positively with plasma beta-carotene concentration. DNA adduct levels of high plasma beta-carotene or alpha-tocopherol groups were not significantly different from the DNA adduct levels of low plasma beta-carotene or alpha-tocopherol groups among current smokers or nonsmokers. In variant states of CYP1A1 or GSTM1 polymorphism, after controlling for effect of cigarettes smoked per day, no significant correlation was found between plasma beta-carotene or alpha-tocopherol and DNA adduct levels. These results indicated that alcohol consumption, cigarette smoking, and plasma alpha-tocopherol have a close relationship with plasma beta-carotene. The plasma beta-carotene and alpha-tocopherol were not likely to influence the level of DNA adducts in lymphocytes.

Postlabeling analysis of DNA adducts in white blood cells from lead smelter workers.

We analyzed DNA adducts levels in white blood cells from 14 lead smelter workers for the first time as an indicator of the effect of lead exposure and compared with some lead exposure indices to evaluate whether lead exposure induces DNA adducts significantly. DNA adducts were measured by the nuclease P1 enhanced 32P-postlabeling method. As the lead exposure indices, we chose blood lead (Pb-B), urinary delta-aminolevulinic acid (ALA) and erythrocyte protoporphyrin (FEP). The levels of DNA adducts had a weak positive correlation with urinary ALA concentration (r = 0.62), but not significant correlation with Pb-B and FEP. This result suggests that lead exposure might have the effect to remain DNA adducts. The inhibition of DNA repair system may be one reason. This preliminary study need be followed by extended surveys on lead exposure.

A unique case of extramammary Paget's disease. Derivation from eccrine porocarcinoma?

A unique case of extramammary Paget's disease is reported that may have derived from eccrine porocarcinoma. A palm-sized erythematous plaque on the patient's pubis spread to the lower abdominal wall. The center of the lesion contained a reddish tumor. Histologic findings of the erythematous plaque showed features of extra-mammary Paget's disease. Those of the reddish tumor, however, corresponded most closely to eccrine porocarcinoma, though we could not entirely rule out that the changes corresponded to larger nests of less differentiated Paget cells. The two distinct neoplastic areas showed continuity both clinically and histologically; our case differed from epidermotropic eccrine porocarcinoma in several clinicopathologic respects. Our case suggests the possibility that extramammary Paget's disease could arise from preexisting porocarcinoma.