Associations between hazard indices of di-n-butylphthalate and di-2-ethylhexylphthalate exposure and serum reproductive hormone levels among occupationally exposed and unexposed Chinese men.

The knowledge on the dose-response relationships between cumulative phthalate exposure and reproductive hormones in human are lacking. To assess the characteristics of the associations between hazard index (HI) of cumulative di-n-butylphthalate (DBP) and di-2-ethylhexylphthalate (DEHP) exposures and serum concentrations of free testosterone (fT), estradiol, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), we used restricted cubic spline function to characterize the dose-response curves between the HI values and reproductive hormones for 74 male workers occupationally exposed to high levels of DBP and DEHP, and 63 male construction workers as comparison group matched for age and smoking status. The median of HI value was 5.30 for exposed workers, 53.0-fold that of unexposed workers (0.10). 89.2% of exposed workers and 1.6% of unexposed workers have HI over 1.00. We observed a borderline significantly negative association between HI and fT in exposed workers (r=-0.195, p=0.096), but not in unexposed workers. The exposed workers showed inverted long-tailed J-shaped fT and FSH curves, and small changes in the LH curve, whereas unexposed workers had inverted and flattened-S-shaped fT and mirror-S-shaped LH and FSH curves. Both T production and hypothalamo-pituitary-testis (HPT) axis function were damaged in workers with high HI of phthalate exposures. HPT feedback function was activated in workers with both high and low HI, and plays an important role in preventing fT level from further decreasing with a rise in HI.

STAT3 is required for the gp130-mediated full activation of the c-myc gene.

The signal transducers and activators of transcription (STAT) family members have been implicated in regulating the growth, differentiation, and death of normal and transformed cells in response to either extracellular stimuli, including cytokines and growth factors, or intracellular tyrosine kinases. c-myc expression is coordinately regulated by multiple signals in these diverse cellular responses. We show that STAT3 mostly mediates the rapid activation of the c-myc gene upon stimulation of the interleukin (IL)-6 receptor or gp130, a signal transducing subunit of the receptor complexes for the IL-6 cytokine family. STAT3 does so most likely by binding to cis-regulatory region(s) of the c-myc gene. We show that STAT3 binds to a region overlapping with the E2F site in the c-myc promoter and this site is critical for the c-myc gene promoter- driven transcriptional activation by IL-6 or gp130 signals. This is the first identification of the linkage between a member of the STAT family and the c-myc gene activation, and also explains how the IL-6 family of cytokines is capable of inducing the expression of the c-myc gene.

Clinical significance of beta 2-microglobulin in serum of adult T cell leukemia.

To clarify the clinical and biological significance of beta 2-microglobulin (beta 2-M) in serum of adult T cell leukemia (ATL) associated with human lymphotropic virus type-I (HTLV-I), beta 2-M was measured in 52 patients with ATL (acute ATL, 35 patients; lymphoma ATL, two patients; chronic ATL, 12 patients; smoldering ATL, three patients), and it was compared with serum lactic dehydrogenase (LDH). Statistical analysis disclosed a correlation between beta 2-M level and the percentage of abnormal lymphocytes (P < 0.05) and platelet count (P < 0.01). There was a correlation between LDH and platelet count (P < 0.01), and a tendency of correlation between LDH and the percentage of abnormal lymphocytes (P < 0.15). Significant difference was present in beta 2-M as well as LDH between acute ATL and chronic ATL (P < 0.01), and between acute ATL and smoldering ATL (P < 0.01). We also investigated a significant inverse correlation between beta 2-M level as well as LDH level and the length of survival after the initial diagnosis (P < 0.01). Thus, the beta 2-M level may indicate the aggressiveness of ATL cells and predict the length of survival.

Urinary excretion of 5-hydroxyindoleacetic acid, delta-aminolevulinic acid and coproporphyrin isomers in rats and men exposed to lead.

The urinary excretion of 5-hydroxyindoleacetic acid (5-HIAA), delta-aminolevulinic acid (ALA) and coproporphyrin (CP) isomers (I and III) was investigated in rats and workers exposed to lead. Urinary excretion of 5-HIAA, which is the major metabolite of serotonin, was not influenced by the lead exposure. However, a significant increase in urinary ALA and CP was found in rats and workers exposed to lead. In particular, the urinary excretion of CP-III was markedly increased compared to that of CP-I in rats and workers exposed to lead.

Comparison of inhibition of erythrocyte pyrimidine 5'-nucleotidase and delta-aminolevulinic acid dehydratase by lead.

The effect of lead on the activities of erythrocyte pyrimidine 5'-nucleotidase (P5N) and delta-aminolevulinic acid dehydratase (ALAD) was investigated in control and lead-exposed workers. The inhibitory effect of lead in vivo and in vitro was more remarkable on the ALAD than on the P5N activity values. It was demonstrated that the pH optimum of both ALAD and P5N in lead workers shifts to the acidic side (nearly pH 6.0) compared to that in control workers.

Effect of lead on the activity of erythrocyte porphobilinogen deaminase in vivo and in vitro.

The effect of lead on the activity of erythrocyte porphobilinogen deaminase (PBGD) in vivo and in vitro was investigated using blood specimens obtained from controls and lead-exposed workers. When lead nitrate was added to the incubation mixture at a final concentration of 10(-4) M, 83% inhibition of erythrocyte PBGD activity was found. However, in workers occupationally exposed to lead, no inhibition of erythrocyte PBGD activity was detected. This finding indicates that the erythrocyte PBGD test is not useful for evaluating exposure to lead in workers. In addition, the in vitro study confirmed that mercuric chloride strongly inhibits erythrocyte PBGD activity.

Tolerance to lead-induced porphyrin metabolic disorders following lead pretreatment in mice.

The protective effect of pretreatment with lead on lead-induced toxicity was investigated in mice, using some biological parameters such as urinary excretion of delta-aminolevulinic acid (ALA) and coproporphyrin, accumulation of erythrocyte protoporphyrin and inhibition of erythrocyte delta-aminolevulinic acid dehydratase; these are useful indicators for evaluating the effects on health of lead. It was demonstrated that pretreatment with a single intraperitoneal dose of 2 mg Pb/kg, 7 days prior to the challenge dose, prevents in part the increasing excretion of urinary ALA induced by a challenge exposure to lead (200 ppm) in the drinking water for 7 days.

Elevated urinary excretion of beta-aminoisobutyric acid and delta-aminolevulinic acid (ALA) and the inhibition of ALA-synthase and ALA-dehydratase activities in both liver and kidney in mice exposed to lead.

Urinary excretion of beta-aminoisobutyric acid (ABA) and delta-aminolevulinic acid (ALA) was investigated in mice exposed to lead (500 p.p.m.) in drinking water for 14 days. Concentrations of both urinary ABA and urinary ALA increased significantly in the lead-exposed mice. However, the degree of increasing excretion was higher in urinary ALA (10-fold of the control) than in urinary ABA (2-fold of the control). On the other hand, it was demonstrated that ALA dehydratase in liver and kidney is inhibited by exposure to lead, while ALA synthase in these tissues has no inhibitory effect.

Species difference of urinary excretion of delta-aminolevulinic acid and coproporphyrin in mice and rats exposed to lead.

The urinary excretion of delta-aminolevulinic acid (ALA) was first investigated in mice exposed to lead in the drinking water (500 ppm) for 30 days, and the results were compared with those obtained from rats exposed to lead under the same exposure condition. The result indicated that the level of urinary ALA excretion in lead-exposed mice was much higher than that in lead-exposed rats. This finding suggests a species difference between mice and rats with regard to sensitivity to lead. On the other hand, urinary coproporphyrin (CP) was also determined in rats and mice exposed to lead. The result obtained indicated that the increasing level of urinary CP excretion was higher in rats than in mice, contrary to expectation. The reason for the discrepancy between urinary excretion of ALA and that of CP in rats and mice exposed to lead is, however, unknown.

The in vitro effect of 13 metals on the activity of erythrocyte pyrimidine 5'-nucleotidase (P5N).

The in vitro effect of 13 metal ions on the activity of erythrocyte pyrimidine 5'-nucleotidase (P5N) was investigated. Particular metals belonging to the boron group have recently been used as materials in a semi-conductor manufacturing. Of these metal ions, Ga exhibited no inhibition of erythrocyte P5N, while in extensively inhibited the erythrocyte P5N activity at concentrations of 10(-4) to 10(-3) M. In addition, the activity of erythrocyte P5N was inhibited by 90-96% when the metal ions such as Cu, Ag, Cd, Hg or Pb were added to the incubation mixture at concentrations of 10(-4) to 10(-3) M. On the other hand, there was no difference between sulfate and nitrate in the in vitro effect of the metal ions on the activity of erythrocyte P5N.

Interrelation between urinary delta-aminolevulinic acid (ALA), serum ALA, and blood lead in workers exposed to lead.

Using a fluorometric HPLC method, we determined delta-aminolevulinic acid (ALA) in sera and urine samples from 16 lead workers with blood lead levels ranging from 19 to 107 microG/100 ml. The concentration of ALA in serum correlated highly with the urinary ALA concentration (gamma = 0.957 for ALA mg/1; gamma = 0.967 for ALA mg/g creatinine). The ALA concentrations in the serum of lead workers ranged from 11 to 151 micrograms/1 with a mean of 51 micrograms/1. In addition, the concentrations of urinary ALA (mg/g creatinine) and serum ALA micrograms/1) had a strong correlation with blood lead concentrations (gamma = 0.838 and 0.892, respectively). These data indicate that the measurement of serum ALA, as well as urinary ALA, is very useful for the biological monitoring of occupational lead exposure.

Measurement of urinary delta-aminolevulinic acid (ALA) by fluorometric HPLC and colorimetric methods.

We devised a fluorometric HPLC method for determining delta-aminolevulinic acid (ALA) in the urine of lead-exposed workers. With this fluorometric HPLC method and the conventional colorimetric method, the concentrations of urinary ALA in 84 lead workers were determined and compared. In the measurement of urinary ALA at lower levels (< or = 5 mg/1), the value of urinary ALA obtained by the fluorometric HPLC method was much lower than that obtained by the conventional colorimetric method, indicating that the colorimetric method also measures urinary ALA-like compounds such as aminoacetone. On the other hand, the measurement of urinary ALA at higher levels (> 5 mg/1) demonstrated that the ALA value obtained by the fluorometric HPLC method corresponded well with that of the conventional colorimetric method. A correlation coefficient between the fluorometric HPLC method and the colorimetric method was 0.856 for 60 urine samples with ALA < or = 5 mg/1, and 0.996 for 24 urine samples with ALA > 5 mg/1.

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.

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.

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.