Benzidine-DNA adduct levels in human peripheral white blood cells significantly correlate with levels in exfoliated urothelial cells.

In a cross-sectional study of 33 workers exposed to benzidine and benzidine dyes and 15 non-exposed controls, we previously reported that exposure status and internal dose of benzidine metabolites were strongly correlated with the levels of specific benzidine-DNA adducts in exfoliated urothelial cells. We also evaluated DNA adduct levels in peripheral white blood cells (WBC) of a subset of 18 exposed workers and 7 controls selected to represent a wide range of adducts in exfoliated urothelial cells. Samples were coded and then DNA was analyzed using 32P-postlabeling, along with n-butanol extraction. One adduct, which co-chromatographed with a synthetic N-(3'-phospho-deoxyguanosin-8-yl)-N'-acetylbenzidine standard, predominated in those samples with adducts present. The median level (range) of this adduct in WBC DNA was 194.4 (3.2-975) RAL x 10(9) in exposed workers and 1.4 (0.1-6.4) in the control subjects (p = 0.0002, Wilcoxon Rank Sum Test). There was a striking correlation between WBC and exfoliated urothelial cell adduct levels (Pearson r = 0.84, p < 0.001) among exposed subjects. In addition, the sum of urinary benzidine, N-acetylbenzidine and N,N'-diacetylbenzidine correlated with the levels of this adduct in both tissues. This is the first study in humans to show a relationship for a specific carcinogen adduct in a surrogate tissue and in urothelial cells, the target for urinary bladder cancer.

Urinary mutagenicity as a biomarker in workers exposed to benzidine: correlation with urinary metabolites and urothelial DNA adducts.

Urinary mutagenicity has been used in occupational and epidemiological studies for over two decades as a cost-effective, general biomarker of exposure to genotoxic agents. However, few studies have compared urinary mutagenicity to additional biomarkers determined among low- and high-exposed groups. To address this issue, we evaluated the relationship between urinary mutagenicity and other types of biomarkers in a cross-sectional study involving 15 workers exposed to the urinary bladder carcinogen benzidine (BZ, high exposure), 15 workers exposed to BZ-dyes (low exposure), and 13 unexposed controls in Ahmedabad, India. Urinary organics were extracted by C18/methanol and evaluated for mutagenicity in the presence of S9 in the Salmonella strain YG1024, which is a frameshift strain that overproduces acetyltransferase. The results were compared to biomarker data reported recently from the same urine samples (Rothman et al., Proc. Natl Acad. Sci. USA, 93, 5084-5089, 1996) that included a metabolite biomarker (the sum of the urinary levels of BZ + N-acetylbenzidine + N,N'-diacetylbenzidine) and a DNA adduct biomarker [a presumptive N-(3'-phosphodeoxyguanosin-8-yl)-N'-acetylbenzidine (C8dG-ABZ) DNA adduct in exfoliated urothelial cells]. The mean +/- SE urinary mutagenicity (revertants/micromol of creatinine) of the low-exposure (BZ-dye) workers was 8.2 +/- 2.4, which was significantly different from the mean of the controls (2.8 +/- 0.7, P = 0.04) as was that of the mean of the high-exposure (BZ) workers (123.2 +/- 26.1, P < 0.0001). Urinary mutagenicity showed strong, positive correlations with urinary metabolites (r = 0.88, P < 0.0001) and the level of the presumptive C8dG-ABZ urothelial DNA adduct (r = 0.59, P = 0.0006). A strong association was found between tobacco use (bidi smoking) and urinary mutagenicity among the controls (r = 0.68, P = 0.01) but not among the exposed workers (r = 0.18, P = 0.11). This study confirms the ability of a biomarker such as urinary mutagenicity to detect low-dose exposures, identify additional genotoxic exposures among the controls, and correlate strongly with urinary metabolites and DNA adducts in the target tissue (urinary bladder epithelia) in humans.

Interaction of Illumination with noise on neuropsychological performance capability.

To determine the interaction effect of illumination with noise performance data were recorded from 20 male college student volunteers on a battery of neuropsychological tests comprising 'memory and search,' 'name and number checking,' 'Flanagan's eye-hand coordination' and 'digit symbol.' Each subject worked under two of the four combinations of illumination (low, 300 lux; high, 500 lux) with quiet (60 dBA) and noise (100 dBA) conditions. A 2 x 2 analysis of variance was performed on the test scores. The accuracy of performance was found enhanced by high illumination in all the tests, but the speed of performance was impaired by noise. The interaction effect was significant, indicating the speed and efficiency of performance increasing while the accuracy of performance affected differently with high illumination under noise condition. However, the error of performance increased by noise under high illumination. Further investigation on a range of neuropsychological tests is suggested before the conclusions are drawn more firmly.

Acidic urine pH is associated with elevated levels of free urinary benzidine and N-acetylbenzidine and urothelial cell DNA adducts in exposed workers.

We evaluated the influence of urine pH on the proportion of urinary benzidine (BZ) and N-acetylbenzidine present in the free, unconjugated state and on exfoliated urothelial cell DNA adduct levels in 32 workers exposed to BZ in India. Postworkshift urine pH was inversely correlated with the proportions of BZ (r = -0.78; P < 0.0001) and N-acetylbenzidine (r = -0.67; P < 0.0001) present as free compounds. Furthermore, the average of each subject's pre- and postworkshift urine pH was negatively associated with the predominant urothelial DNA adduct (P = 0.0037, adjusted for urinary BZ and metabolites), which has been shown to cochromatograph with a N-(3'-phosphodeoxyguanosin-8-yl)-N'-acetylbenzidine adduct standard. Controlling for internal dose, individuals with urine pH < 6 had 10-fold higher DNA adduct levels compared to subjects with urine pH > or = 7. As reported previously, polymorphisms in NAT1, NAT2, and GSTM1 had no impact on DNA adduct levels. This is the first study to demonstrate that urine pH has a strong influence on the presence of free urinary aromatic amine compounds and on urothelial cell DNA adduct levels in exposed humans. Because there is evidence that acidic urine has a similar influence on aromatic amines derived from cigarette smoke, urine pH, which is influenced by diet, may be an important susceptibility factor for bladder cancer caused by tobacco in the general population.

The glutathione S-transferase M1 (GSTM1) null genotype and benzidine-associated bladder cancer, urine mutagenicity, and exfoliated urothelial cell DNA adducts.

Multiple studies in the general population have suggested that subjects with the glutathione S-transferase M1 (GSTM1)-null genotype, who lack functional GSTM1, are at higher risk for bladder cancer. To evaluate the impact of the GSTM1-null genotype on bladder cancer caused by occupational exposure to benzidine and to determine its influence on benzidine metabolism, we carried out three complementary investigations: a case-control study of bladder cancer among workers previously exposed to benzidine in China, a cross-sectional study of urothelial cell DNA adducts and urinary mutagenicity in workers currently exposed to benzidine in India, and a laboratory study of the ability of human GSTM1 to conjugate benzidine and its known metabolites in vitro. There was no overall increase in bladder cancer risk for the GSTM1-null genotype among 38 bladder cancer cases and 43 controls (odds ratio, 1.0; 95% confidence interval, 0.4-2.7), although there was some indication that highly exposed workers with the GSTM1-null genotype were at greater risk of bladder cancer compared to similarly exposed workers without this allele. However, the GSTM1 genotype had no impact on urothelial cell DNA adduct and urinary mutagenicity levels in workers currently exposed to benzidine. Furthermore, human GSTM1 did not conjugate benzidine or its metabolites. These results led us to conclude that the GSTM1-null genotype does not have an impact on bladder cancer caused by benzidine, providing a contrast to its association with elevated bladder cancer risk in the general population.

The impact of interindividual variation in NAT2 activity on benzidine urinary metabolites and urothelial DNA adducts in exposed workers.

Several epidemiologic studies indicate that NAT2-related slow N-acetylation increases bladder cancer risk among workers exposed to aromatic amines, presumably because N-acetylation is important for the detoxification of these compounds. Previously, we showed that NAT2 polymorphisms did not influence bladder cancer risk among Chinese workers exposed exclusively to benzidine (BZ), suggesting that NAT2 N-acetylation is not a critical detoxifying pathway for this aromatic amine. To evaluate the biologic plausibility of this finding, we carried out a cross-sectional study of 33 workers exposed to BZ and 15 unexposed controls in Ahmedabad, India, to evaluate the presence of BZ-related DNA adducts in exfoliated urothelial cells, the excretion pattern of BZ metabolites, and the impact of NAT2 activity on these outcomes. Four DNA adducts were significantly elevated in exposed workers compared to controls; of these, the predominant adduct cochromatographed with a synthetic N-(3'- phosphodeoxyguanosin-8-yl)-N'-acetylbenzidine standard and was the only adduct that was significantly associated with total BZ urinary metabolites (r = 0.68, P < 0.0001). To our knowledge this is the first report to show that BZ forms DNA adducts in exfoliated urothelial cells of exposed humans and that the predominant adduct formed is N-acetylated, supporting the concept that monofunctional acetylation is an activation, rather than a detoxification, step for BZ. However, because almost all BZ-related metabolites measured in the urine of exposed workers were acetylated among slow, as well as rapid, acetylators (mean +/- SD 95 +/- 1.9% vs. 97 +/- 1.6%, respectively) and NAT2 activity did not affect the levels of any DNA adduct measured, it is unlikely that interindividual variation in NAT2 function is relevant for BZ-associated bladder carcinogenesis.

The correlation of chest radiograph and pulmonary function tests in asbestos miners and millers.

An environmental-cum-medical survey was carried out in asbestos mines and milling units at Pullivendalla, Cuddaph (A.P.) India. This was done in two mines and six milling units with 95% of the total work force being surveyed. Out of a total of 633 registered workers, 329 (52%, all males) were employed in mines while 135 (21.4%) workers of whom 114 (84%, all females) were employed in the milling units. All subjects underwent limited medical examination, spirometry and chest radiographs. The levels of asbestos fiber concentration was much below threshold limit value (TLV) in underground mines but several times higher than TLV in milling units. The percentage of workers with abnormal pulmonary function tests (PFT) and chest radiographs increased with duration of exposure in smokers as well as non-smokers. Restrictive pattern of lung functions (159 workers-16.27%) was more common than obstructive (33 workers-5.21%) and combined type (22 workers-3.4%). Similarly, the parenchymal changes (156 workers-24.6%) were more common than pleural (27 workers-4.3%). As most of the males were employed in mines, where the fiber levels were much below TLV, the number of male workers with normal PFT and chest radiographs were ten times (61.3%) more than male workers with both the parameters abnormal (6.3%). As most of the females (114 our of 120--95%) were employed in milling units, where the levels of fibers were several times higher than TLV, the number of females having both the parameters normal 29 (24.1%) or abnormal 35 (29.2%) were similar.

Determination of benzidine, N-acetylbenzidine, and N,N'-diacetylbenzidine in human urine by capillary gas chromatography/negative ion chemical ionization mass spectrometry.

We report an isotope dilution mass spectrometry method using capillary gas chromatography/negative ion chemical ionization to quantitate urine concentrations of benzidine (BZ) and of its acetylated metabolites N-acetylbenzidine (ABZ) and N,N'-diacetylbenzidine (DABZ). Urine samples were purified by solid-phase extraction columns, reduced with LiAlH4/THF, and derivatized with pentafluoropropionic anhydride. The derivatives were measured by selected ion monitoring relative to deuterium-labeled internal standards. A detection limit as low as 0.5, 0.8, and 1.5 ppt for BZ, ABZ, and DABZ, respectively, can easily be achieved. Urinary concentrations of ABZ substantially exceed those of either BZ or of DABZ in workers exposed to BZ or BZ-based dyes. This method has been successfully used to measure BZ, ABZ, and DABZ in 1.0-ml urine samples collected from workers involved in manufacturing BZ and BZ-based dyes. The method should be applicable to the measurement of other aromatic amines and their acetylated metabolites.

Calcutta metro: is it safe from noise pollution hazards?

A modest assessment of noise was made in Calcutta Metro, India's first ever underground tube rail system, to examine if the range of noise levels present could endanger the hearing sensitivity of workers for the Metro. Sound measuring instruments of a sound level meter, an octave band analyzer, and a sound level calibrator were used for measuring the sound pressure levels in platforms of three stations: Esplanade, Kalighat and Tollygunge. The results indicated that the averaged A-weighted SPLs in these stations were in the range of 84-87 dBA. In the coaches of the moving train the Leq values ranged 92-99 dBA and LNP 105-117 dBA, all exceeding the safe limit of day time noise exposure of 55 dBA and 85 dBA of ACGIH. The SPLs at 4,000 Hz in the coaches were also in excess of safe exposure limit of 79 dB. The findings thus posed a potential threat to the workers.

Neurobehavioural changes in workshop painters of a public transport network.

To determine alterations in functional capacities due to exposure to solvents in painters, a battery of psychological tests consisting of associative recall, critical flicker fusion, letter cancellation, Muller-Lyer illusion and card sorting was administered before their work day schedule starting at 0800 h, 16 h after cessation of their work the previous day. Simultaneously a reference group of diesel worker and a control group were also tested at the same time of day. Age-adjusted performance scores were calculated for the painter group and diesel worker group based on the linear age trend of the control group. Analysis of variance suggested significant degradation in performance on tests of critical flicker fusion (alternate), letter cancellation, Muller-Lyer illusion and card sorting (by design configuration). Persistence of solvent exposure effects through visual and motor function impairments is reflected.

Dustiness, silicosis & tuberculosis in small scale pottery workers.

Studies were carried out in eight small scale potteries to find out the airborne dust concentrations and the prevalence of dust related diseases like silicosis and tuberculosis in 292 workers. Chest radiography revealed that 44 (15.1%) pottery workers were suffering from silicosis and an equal number showed radiological evidence of tuberculosis. The environmental study showed that the concentrations of airborne dust, containing free silica, in the work environment of all departments (except packing department) of potteries were higher than threshold limit values (TLVs). The prevalence of silicosis and tuberculosis correlated with the levels of airborne dust. The prevalence of tuberculosis increased with radiological severity of silicosis. Dust control measures combined with pre-employment and periodical medical examinations are recommended for the control of silicosis and tuberculosis in the pottery industry.

Behavioral effects and body activity level in female hospital staff nurses during work hour.

To examine if there are changes in the levels of affections over different times of day, affective state measures, alertness, irritability, mental tension, headache and anxiety of 100 female staff nurses were assessed, on visual analogue scales at 2-h intervals over a 24-h work cycle spread over three different shift works. Oral temperature was also recorded simultaneously. Sleep pattern, dietary habit and menstruation cycle of subjects were monitored. Analysis of variance reflected a significant rise in oral temperature as the day progressed with a peak during 16:00-18:00 h, which however gradually declined thereafter. Significant changes in the affective states indicated ascendancy in alertness, mental tension and anxiety within the first few hours of the beginning of each shift work, but descendancy towards the end of it. Further studies on some more affective states and physiological measures which may show parallelism have been suggested.

Assessment of noise environment in a major railway station in India.

To assess the degree of noise pollution in relation to the health and safety of the employees and commuters, a study on the levels of noise originating from various locomotives and also from different other sources was undertaken at Kalupur railway station of Gujarat State in India. The sound pressure levels (SPL) were measured on platforms. Noise dose count was monitored on a ticket collector on duty for 8 h. The results reflected that the SPLs in the platforms well exceeded the day time noise exposure limit. Further, the SPLs produced by loudspeakers were high and those by train whistles were intensely high pitched. The noise dose count of the ticket collector was also in excess of ACGIH standard. All these suggest that the SPLs in the platforms can endanger the healthful living of the users. Suitable control measures have been suggested.

Assessment of location-specific human exposure to dichloro-diphenyl trichloroethane and benzenehexachloride in Gujarat state, India.

On the basis of the use of insecticides in agriculture and vector control programmes, two locations were selected in Gujarat state, India. In location 1 the insecticides are used in both agriculture and vector control programmes while in location 2 they are used only in agriculture. Raw food commodities, water, soil and blood samples were collected from the people residing in these locations, and analysed for total dichloro-diphenyl trichloroethane and total benzenehexachloride residues. Residue levels were significantly lower in location 2 than in location 1.

Assessment of comfort of various hearing protection devices (HPD).

To evaluate the comfort of hearing protection devices, two models of ear plugs and five models of ear muffs were tested. The psychophysical method of 'single stimuli' was applied on a group of 30 subjects with or without wearing the devices for a short duration of 15 min under noise condition of 100 dBa in the acoustic chamber as also on a group of 10 weavers with the protection devices worn for longer durations of 1 h, 4 h and 8 h under noise exposure of 102-104 dBA in the weaving shed. Each subject performed 8 trials with each type of device on different days. Application force and tightness of spring were also evaluated. The results yielded a comfort grading for hearing protection devices. The comfort grading, however, depended on several factors in addition to application force and tightness of spring, which has been discussed.

Humoral immunological profile of workers exposed to asbestos in asbestos mines.

Humoral immunological profile including immunoglobulins IgG, IgA, IgM, C-reactive protein, rheumatoid factor, antinuclear antibodies and circulating immune complexes were studied in a representative sample of 36 workers suffering from asbestosis (group A), 35 workers who are exposed to asbestos but not having evidence of asbestosis (group B) and 28 control workers (group C). Mean IgG and IgA levels were found to be significantly higher in the two exposed groups than in the controls. Circulating immune complexes of IgG, IgA and IgM class were detected in a significant percentage of cases in exposed groups than in controls. In groups A and B, the percentage of positive ANF cases was much higher than in the controls. The results suggest that immunological changes are associated with exposure to asbestos and these may play an important role in the pathogenesis of the disease process.

Evaluation of human exposure to the persistent insecticides DDT and HCH in Ahmedabad, India.

Concentrations of two organochlorine insecticides, DDT and HCH, in 21 human adipose tissue samples (from cadavers) and 20 venous blood samples collected in Ahmedabad, India were determined. None of the cases had any history of occupational exposure to organochlorine insecticides. Residue concentrations of DDT and HCH in these samples indicated considerable exposure of the general population to these chemicals. Samples from males had comparatively higher concentrations of these insecticide residues than females. The mean residue concentrations of total DDT and HCH in the blood serum of males was 213.83 and 70.051 ppb, respectively, whereas females had DDT and HCH concentrations of 177.38 and 65.44 ppb, respectively. Total DDT and HCH in adipose tissue was 3.967 and 4.054 ppm in males, and 3.538 and 3.144 ppm in females, respectively. Residue concentrations reported here are lower than those reported earlier.

Pulmonary function tests--a comparison between urban industrial and rural agricultural workers of Andhra Pradesh.

Physical parameters and pulmonary function tests (VC, FEV1, FEV1% and FEF25-75%) were collected in 112 urban industrial workers and 104 rural agricultural workers. These average values, values according to age and smoking habits were compared between urban and rural workers. Inspite of no differences in age, rural workers average height, weight, BSA is significantly lower than urban workers. A significant increase in VC value to the extent of 0.22 lit (7.1%), significantly lower FEV 1% value by 4.7% and reduce FEF25-75% value (4.7%) is demonstrated in rural than urban workers. Smoking is shown to produce airway obstruction in both urban and rural workers. A wide variation of PFT values between the present workers and other reported values in India is observed. This study indicates rural workers have better pulmonary capacity and less flow rates than their urban counterpart.