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Urinary biomarkers of occupational N,N-dimethylformamide (DMF) exposure attributed to the dermal exposure.

OBJECTIVE: To evaluate whether the dermal exposure to N,N-dimethylformamide (DMF) exerts significant effects and to determine the unit increment of dermal exposure on the total body burden of two biomarkers in urine: metabolism-required N-methylformamide (U-NMF) and non-metabolized DMF (U-DMF) in actual occupational environments. METHODS: Exposure via respiratory and dermal routes was assessed on an individual basis for 75 workers from four DMF-related factories directly exposed to DMF. Respiratory exposure was determined by breathing-zone sampling for a full-work shift, and dermal exposure was assessed on the palms and forearms of both hands by an adhesive tape-patch method. U-NMF and U-DMF collected immediately postshift were measured. RESULTS: The average concentrations of airborne DMF, DMF on hands and on forearms, U-NMF, and U-DMF (GM) were 1.51 ppm, 0.04 microg/cm(2), 0.03 microg/cm(2), 0.47 mg/l, and 0.38 mg/l, respectively. In multiple linear regression tests, only airborne DMF and DMF on hands remained significantly (P<0.001) associated with U-NMF and U-DMF. Based on model estimates, the unit increment of hands' exposure (microg/cm(2)) could contribute to 0.53 and 0.46 mg/l of the increment of U-NMF and U-DMF, respectively, given a daily occupational airborne exposure to DMF at about 1.5 ppm. CONCLUSIONS: Dermal exposure provides a substantial contribution to the total body burden of DMF. A control remedy such as the enforcement of wearing impermeable gloves by workers occupationally exposed to DMF should be implemented with the highest priority.

Administration, Cutaneous↗

The methyl C-H blueshift in N,N-dimethylformamide-water mixtures probed by two-dimensional Fourier-transform infrared spectroscopy.

Two-dimensional correlation spectroscopy was used to study the composition-dependent spectral variations of the CH-stretching bands of N,N-dimethylformamide (DMF)-water mixtures with X(DMF) ranging from 0.98 to 0.60. By a detailed correlation analysis of the spectral changes of the CH- and OH-stretching bands, it is found that the intensities of the CH and OH bands change in different ways when the water content is increased. It is also found that two different regions of the water content can be distinguished, in which the intensity changes have different signatures. A tentative explanation for how these phenomena might be related to structural changes in the mixture is proposed. The structural change of DMF induced by the water hydrogen bonded on the carbonyl group is supposed to be the possible origin of the methyl C-H blueshift instead of the direct C-H...O interactions before the hydrophobic hydration takes place.

Algorithms↗

Effects of dimethylformamide (DMF) on coagulation and platelet activity.

The effects of dimethylformamide (DMF) on hemostatic functions, especially on platelet activity, were examined both in vitro and in vivo in 15 workers exposed to DMF (27 mg/m3, median value). Twenty-eight control subjects who were not exposed to DMF, but comparable for age, anthropometric data, and smoking habits, were also studied. Workers exposed to DMF showed a decrease in the number of platelets and had longer coagulation times, probably due to a change caused by DMF on the membrane receptor of platelets and on the phospholipid components of the clotting system.

Adenosine Diphosphate↗

Dimethylformamide-induced liver damage among synthetic leather workers.

Prevalence of liver injury associated with dimethylformamide (DMF) exposure was determined. Medical examinations, liver function tests, and creatine phosphokinase (CPK) determinations were performed on 183 of 204 (76%) employees of a synthetic leather factory. Air concentrations of solvents were measured with personal samplers and gas chromatography. The concentration of DMF in air to which each worker was exposed was categorized. High exposure concentrations of DMF (i.e., 25-60 ppm) were significantly associated with elevated alanine aminotransferase (ALT) levels (ALT greater than or equal to 35 IU/l), a result that did not change even after stratification by hepatitis B carrier status. Modeling by logistic regression demonstrated that exposure to high concentrations of DMF was associated with an elevated ALT (p = .01), whereas hepatitis B surface antigen (HBsAg) was slightly but independently associated with an elevated ALT (p = .07). In those workers who had normal ALT values, there occurred still significantly higher mean ALT and aspartate aminotransferase (AST) activities, especially among those who were not HBsAg carriers. A significant association existed between elevated CPK levels and exposure to DMF. However, an analysis of the CPK isoenzyme among 143 workers did not reveal any specific damage to muscles. This outbreak of liver injury among synthetic leather workers is ascribed to DMF. It is recommended that the occupational standard for DMF and its toxicity among HBsAg carriers be evaluated further.

Adult↗

Monitoring for N,N-dimethylformamide and N,N-dimethylacetamide with a diffusive sampler using distilled water as an absorbent.

To obtain an appropriate sampling method for evaluating individual workers' exposure concentrations to chemicals highly soluble in water, the suitability of LiPS (liquid passive sampler) as diffusive sampler was examined. Although adsorbed chemical substances with activated carbon felt being used as the adsorbent must be desorbed in such samplers, this process can be omitted by using liquid as the absorbent. This kind of simplification can be utilized for water-soluble solvents such as N,N-dimethylformamide (DMF) and N,N-dimethylacetamide (DMAC). In laboratory experiments DMF and DMAC levels determined by this method were sufficiently correlated with exposure concentrations, exposure duration, and cumulative exposure levels. The sampling rates for DMF and DMAC obtained from these results were 0.50 and 0.43 mL/min, respectively, and the theoretical sampling rate for DMF obtained using Fick's diffusion formula was consistent with the experimental value. The effects of relative humidity, wind velocity, and temperature on the measured values were negligible within the range expected in a practical industrial environment. In a field survey the exposure concentrations for 38 workers handling DMF in the process of manufacturing spandex fiber and polyurethane gloves were determined both by this method and by the National Institute of Occupational Safety and Health (NIOSH) method, in which silica gel tubes are used. The DMF concentrations detected by the two methods were comparable (Y = 0.912X, correlation coefficient: 0.932). This LiPS method proved to be useful in determining the concentrations of DMF and DMAC to which workers were exposed.

Absorption↗

Direct automated EMIT d.a.u. analysis of N,N-dimethylformamide-modified serum, plasma, and postmortem blood for benzodiazepines, benzoylecgonine, cannabinoids, and opiates.

Serum, plasma, and postmortem blood treated with two volumes of N,N-dimethylformamide (DMF) and centrifuged, were directly analyzable by means of the EMIT d.a.u. reagents on the Syva Autocarousel in the same manner as urine. Cutoff values in milligrams (immunochemically cross-reactive analyte equivalents)/L are 0.05 for morphine (MOR), 0.15 for benzoylecgonine (BZE), 0.20 for oxazepam (OX), and 0.02 for 11-nor-delta 9-tetrahydrocannabinol carboxylic acid (THCC). The relationship of concentrations (micrograms/mL serum) to absorbance changes (delta A) were S-shaped up to greater than 3.0 for MOR, 4.0 for BZE, greater than 5.0 for OX, and 0.2 for THCC. Beyond these maximal concentrations, delta A values declined. Thus, negatives should be repeated on substantially diluted aliquots to avoid missing extraordinarily high positives. "False" quantitative negative/positive noncongruence between total EMIT cross-reactives and free-drug analyses by gas chromatography/mass spectrometry (GC/MS) were 0/17 (N = 75) for opiates, 8/0 (N = 119) for cocaine products, and 19/5 (N = 103) for cannabinoids. For benzodiazepines (N = 58) the "false" negative/"false" positive ratio of EMIT (total)/high performance liquid chromatography (HPLC) (free) was 4/4. Within-day precision as coefficient of variation (CV) of quantitative estimates was 8-18%. For between-day precision, quantitative estimates varied by 8% for MOR, 15% for BZE, 18% for OX, and 34% for THCC.

Benzodiazepines↗

Direct automated EMIT d.a.u. analysis of N,N-dimethylformamide-modified serum, plasma, and postmortem blood for amphetamines, barbiturates, methadone, methaqualone, phencyclidine, and propoxyphene.

The addition of two volumes of N,N-dimethylformamide (DMF) to serum, plasma, and postmortem blood with subsequent centrifugation resulted in supernatant that could be directly analyzed by EMIT d.a.u. urine reagents on the Syva autocarousel. Application of this method to the drugs below gave cutoff concentrations in milligrams of immunochemically cross-reactive analyte equivalents/L as follows: 0.05 for amphetamine, 0.05 for secobarbital, 0.075 for methadone, 0.05 for methaqualone, 0.025 for phencyclidine, and 0.05 for propoxyphene. Quantitative "false" negative/positive noncongruence between total EMIT cross-reactants and free-drug analyses by gas chromatography/mass spectrometry were 3/4 (n = 50) for amphetamines, 2/0 (n = 60) for barbiturates, 0/0 (n = 47) for methadone, 0/0 (n = 48) for methaqualone, 1/0 (n = 44) for phencyclidine, and 1/2 for propoxyphene (n = 53). Within-day precision, as indicated by the coefficient of variation, of quantitative estimates using low and high controls ranged from 3.7 to 11% and 1.8 to 10.3%, respectively. Using the same control levels, between-day precision of quantitative estimates varied from 5.8 to 30.3% and 3.0 to 11.8%, respectively.

Amphetamines↗

Biological monitoring of workers exposed to dimethylformamide in a textile polyurethane unit.

Dimethylformamide (DMF) is a universal solvent noted for its solubility in both aqueous and lipid media. It is hepatotoxic, and there are concerns about its carcinogenicity. Our objective was to determine the correlation between air monitoring measurements and biological measurements of a suitable metabolite of DMF in a cohort of operatives in a polyurethane production unit. This was done with a view to assessing how much the inhalation route contributed to total DMF exposure, mainly for control purposes. We investigated the relationship between personal air sample measurements of DMF and biological measurements of N-methylformamide (NMF) in nine adult subjects, recruited across the shifts, with varying levels of exposure to DMF. Personal exposure monitoring was carried out with a low-flow-rate Model 222-4 SKC pump, while post-shift urine samples were obtained for further analysis. Operatives were asked to abstain from consuming alcohol for 24 h before the designated shift, as advised by the laboratory responsible for the analysis of urine samples. We found a very strong statistical association between air sample measurements of DMF and NMF in the urine of the sample population (R(2) = 0.95, P < 0.0001). This study suggests that airborne exposure contributes significantly to elevated levels of NMF and, therefore, indicates that efforts should be concentrated on engineering controls in reducing workplace DMF exposure.

Adult↗

Directed evolution of subtilisin E in Bacillus subtilis to enhance total activity in aqueous dimethylformamide.

Sequential rounds of error-prone PCR to introduce random mutations and screening of the resultant mutant libraries have been used to enhance the total catalytic activity of subtilisin E significantly in a non-natural environment, aqueous dimethylformamide (DMF). Seven DNA substitutions coding for three new amino acid substitutions were identified in a mutant isolated after two additional generations of directed evolution carried out on 10M subtilisin E, previously "evolved' to increase its specific activity in DMF. A Bacillus subtilis-Escherichia coli shuttle vector was developed in order to increase the size of the mutant library that could be established in B.subtilis and the stringency of the screening process was increased to reflect total as well as specific activity. This directed evolution approach has been extremely effective for improving enzyme activity in a non-natural environment: the resulting-evolved 13M subtilisin exhibits specific catalytic efficiency towards the hydrolysis of a peptide substrate succinyl-Ala-Ala-Pro-Phe-p-nitroanilide in 60% DMF solution that is three times that of the parent 10M and 471 times that of wild type subtilisin E. The total activity of the 13M culture supernatant is enhanced 16-fold over that of the parent 10M.

Anilides↗

Thirteen-week inhalation toxicity of N,N-dimethylformamide in F344/N rats and B6C3F1 mice.

Male and female F-344 rats and B6C3F1 mice (10/sex/group) were exposed to N,N-dimethylformamide (DMF) by whole body inhalation exposure at 0, 50, 100, 200, 400, or 800 ppm, 6 h/day, 5 days/week, for 13 weeks. A concentration-dependent depression in body weight occurred in rats of both sexes at 400 (6-11%) and 800 ppm (20-22%). In contrast, all weight changes in both sexes of mice were within 10% of controls. No rats died, while 5 mice died from nonexposure-related causes. Relative liver weights were significantly increased at all DMF concentrations in both sexes and both species. Activities of serum sorbitol dehydrogenase (SDH) were statistically increased in male and female rats (200 to 800 ppm) on study days 4, 24, and 91 (13 weeks). Activities of alanine aminotransferase (ALT) and isocitrate dehydrogenase (ICD) were statistically increased in both sexes of rats exposed to 800 ppm DMF at all time points. Cholesterol (CHOL) levels were statistically increased in male and female rats (50-800 ppm) at all sampling time points. Levels of total bile acids (TBA) were statistically increased in both sexes of rats (400-800 ppm) on days 24 and 91. Centrilobular hepatocellular necrosis (minimal to moderate) was seen in rats of both sexes exposed at 400 and 800 ppm, with the lesions more severe in females. Centrilobular hepatocellular hypertrophy (minimal to mild) was found in all groups of DMF-exposed male mice, and in female mice exposed at 100-800 ppm. For male and female rats the no-observed-adverse-effect concentration (NOAEC) for microscopic liver injury was 200 ppm. The NOAEC was 50 ppm for female mice, but an NOAEC based upon the absence of microscopic liver injury was not determined in male mice.

Administration, Inhalation↗

Growth-inhibitory and differentiation-inducing activity of dimethylformamide in cultured human malignant glioma cells.

OBJECTIVE: To determine the growth-inhibitory and differentiation-inducing activity of dimethylformamide (DMF) on a human glioma cell line (SHG-44). DMF is a type of polar solvent and a potent differentiation-inducing agent in many kinds of human solid tumors, yet its effect on human glioma remains unclear. METHODS: The effects of DMF on cell proliferation using 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, cell cycle distribution (with flow cytometry), colony-forming efficiency in double-layer soft agar, tumorigenicity in athymic nude mice, morphological changes, and glial fibrillary acidic protein expression were studied. RESULTS: At dose ranges of 0.25, 0.5, 0.75, and 1.0%, DMF caused a dose-dependent proliferation inhibitory effect in monolayers and a marked dose-dependent suppression of colony-forming efficiency in double-layer soft agar with a complete loss of colony-forming ability in cells exposed to 0.75 and 1.0% DMF. Accumulation of cells in G0/G1 phases was observed in DMF-treated (0.5 and 1.0%) cells, also in a dose-dependent manner. SHG-44 cells exposed to DMF (0.5 and 1.0%) for 15 days changed morphologically from small spindle-shaped to large polygonal and flattened stellate cells with multiple slender processes. These cells were still tumorigenic in athymic nude mice, but the growth of xenografts was remarkably reduced, especially in the 1.0% DMF-treated group. The expression of glial fibrillary acidic protein was notably increased by DMF (0.5 and 1.0%). Washout experiments revealed that the effects of DMF on cell proliferation and cell cycle distribution were reversible. CONCLUSION: Our results suggest that DMF drove the SHG-44 cells to a more mature phenotype with inhibited growth.

Adult↗

Cancer incidence of workers exposed to dimethylformamide and/or acrylonitrile.

This study was to determine whether exposure to dimethylformamide (DMF) and acrylonitrile (ACN) separately or in combination, was associated with increased cancer incidence. Workers exposed to DMF and/or ACN were observed from 1956 through 1984 for cancer incidence. The workers exposed to DMF but not ACN showed significant excesses in incidence for buccal cavity and pharynx cancer and malignant melanoma. A significant excess of prostate cancer incidence was observed among workers exposed to DMF and ACN. No dose-response relationships were observed between DMF or ACN exposure and cancer incidence. The significant excesses in cancer incidence among employees exposed to DMF and/or ACN could be due to statistical chance or other factors, such as tobacco and alcohol consumption. Further studies are warranted.

Acrylonitrile↗

Mortality study of workers exposed to dimethylformamide and/or acrylonitrile.

Workers exposed to dimethylformamide (DMF) and/or acrylonitrile (ACN) were observed from 1950 through 1982 for mortality. The wage-roll workers exposed to DMF showed significant excess in total deaths attributable mainly to ischemic heart disease and external causes when compared with rates from E. I. Du Pont de Nemours & Co. However, there were no significant excesses in mortality when expected numbers were based on US or local statistics. No dose-response relationships were observed between DMF or ACN exposure and mortality. The significant excesses in mortality among employees exposed to DMF and/or ACN could be due to statistical chance or life-style factors

Acrylonitrile↗

Abnormal liver function associated with occupational exposure to dimethylformamide and hepatitis B virus.

N,N-Dimethylformamide (DMF) has excellent solvent properties and is used intensively in the production of synthetic leather and resins. It has caused hepatoxicity in human and animal studies. Hepatitis B virus (HBV) and hepatitis C virus infections are reported to be the major causes of chronic liver diseases (including liver cirrhosis and liver cancer) in Taiwan. This study examined the dose-response relationship of the observed abnormal liver function among the DMF-exposed workers and the interactions among DMF, other chemical exposures, HBV infection, and potential confounders on liver abnormalities. The average DMF exposure concentration was 11.6 ppm (median, 5.9 ppm; range, 0.1 to 86.6 ppm); 65 of 176 workers (36.9%) had high (> 10 ppm) DMF exposure, 37 (21%) had middle (> 5 ppm, < or = 10 ppm) exposure, and 74 (42%) had low (< or = 5 ppm) exposure. There were 24 of 65 abnormal liver function test results (LFTs) (36.9%) (elevations of either glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, or gamma-glutamyl transpeptidase) among the workers with high DMF exposure, 10 of 37 abnormal LFTs (27%) among workers with middle DMF exposure, and 11 of 74 abnormal LFTs (22%) among workers with low DMF exposure. Compared with the workers having low DMF exposure, the HBV, drinking, body mass index (BMI), sex, duration of employment, epichlorohydrin, and toluene exposure adjusted odds ratios (ORs) (and 95% confidence intervals [CIs]) for abnormal LFTs were 1.62 (0.61, 4.28) for workers with middle DMF exposure and 2.93 (1.27, 6.8) for those with high DMF exposure, and there was a significant dose response between DMF exposure and the prevalence of abnormal LFTs (P = 0.006). There were significant associations between abnormal LFTs and HBV carriers (adjusted OR: 3.11; 95% CI: 1.29, 7.5; P = 0.01) and between abnormal LFTs and increased BMI (adjusted OR: 2.2; 95% CI: 1.02, 4.72; P = 0.041). Ultrasonography showed significant associations between chronic liver diseases and HBV carrier status, increased BMI, and high cumulative (> 100 ppm-years) DMF exposure (respectively, adjusted OR: 9.58, 95% CI: 1.79, 51.4, P = 0.007; adjusted OR: 13.2, 95% CI: 1.32, 132, P = 0.025; and adjusted OR: 6.2, 95% CI: 1.14, 34.1, P = 0.032). Drinking and BMI were significantly associated with fatty liver (respectively, adjusted OR: 4.9, 95% CI: 1.39, 17.3, P = 0.012; and adjusted OR: 7.93, 95% CI: 1.6, 39.3, P = 0.01). In conclusion, this study demonstrated that (1) a significant dose-response relationship existed between liver function abnormalities and DMF exposure among workers in Taiwan, (2) HBV carrier status or increased BMI had synergistic effects with DMF in causing liver abnormalities (abnormal LFTs and clinical chronic liver diseases).

Adult↗

Evaluation of current biological exposure index for occupational N, N-dimethylformamide exposure from synthetic leather workers.

The aim of this study was (1) to investigate the correlation between external exposure to N, N-dimethylformamide (DMF) and urinary excretion of DMF and N-methylformamide; (2) to assess whether the correspondence between the current occupational exposure limit setting and recommended urinary biological exposure index is substantial; and (3) to evaluate whether coexposure to toluene, methyl ethyl ketone, and ethyl acetate has an effect on urinary excretion of DMF and N-methylformamide (NMF). Urinary DMF and NMF were significantly correlated (P < 0.01) with one another and also significantly correlated with airborne DMF (P < 0.01) over the range of 1.55 to 152.8 mg/m. Urinary DMF can be considered a complementary marker for short-term exposure. Urinary concentration of NMF and DMF, corresponding to the 8-hour exposure to airborne DMF at 30 mg/m, was estimated to 38.4 mg/L or 39.4 mg/g creatinine for NMF and to 0.92 mg/L or 0.96 mg/g creatinine for DMF.

Acetates↗

Paracoccus aminophilus sp. nov. and Paracoccus aminovorans sp. nov., which utilize N,N-dimethylformamide.

Two methylamine- and N,N-dimethylformamide-utilizing Paracoccus spp. are described. These bacteria are gram-negative, nonsporeforming, nonmotile, coccoid or short rod-shaped organisms. Their DNA base composition is 62 to 68 mol% G + C. Their cellular fatty acids include large amounts of C18:1 acid. Their major hydroxy acids are 3-OH C10:0 and 3-OH C14:0 acids. The major ubiquinone is Q-10. These bacteria are distinguished from Paracoccus denitrificans and Paracoccus alcaliphilus by physiological characteristics and by DNA-DNA-homology. Paracoccus aminophilus sp. nov. and Paracoccus aminovorans sp. nov. are proposed. The type strain of P. aminophilus is DM-15 (= JCM 7686), and the type strain of P. aminovorans is DM-82 (= JCM 7685). Paracoccus halodenitrificans is distinguished from other Paracoccus species on the basis of cellular fatty acid composition, hydroxy fatty acid composition, and DNA-DNA homology. It may not be a valid member of the genus Paracoccus.

Base Composition↗

(5,11,17,23-Tetra-tert-butyl-26,28-dihydroxycalix[4]arene-25,27-dioxy)diacetic acid N,N-dimethylformamide trisolvate.

The title compound, C48H60O8.3C3H7NO, is a derivative of calix[4]arene in the cone conformation, modified with distal carboxylic acid functional groups at the lower rim. A clathrate dimethylformamide (DMF) molecule is located within the calix[4]arene cone, and two DMF solvate molecules participate in hydrogen bonding with the carboxylic acid groups. Intramolecular hydrogen bonds are also formed between the OH groups and the adjacent ether groups in the partially substituted calix[4]arene.

Calixarenes↗

Mixtures of trifluoroethanol or hexafluoroisopropanol and dimethylformamide are not of general applicability for peptide condensations catalyzed by trypsin.

Mixtures of a good hydrogen bond donor, 2,2,2-trifluoroethanol (TFE) or 1,1,1,3,3,3-hexafluroisopropanol, and an acceptor, dimethylformamide (DMF) (1:1, v/v), containing 4% buffer have been described as adequate solvent systems for trypsin-catalyzed peptide fragment condensations [Mihara et al. (1993) Int. J. Pept. Protein Res. 41, 405]. Thus, we decided to study the behaviour of trypsin in such solvent systems. We investigated whether this protease would efficiently catalyze condensations between fragments derived from an analogue of the gp-41 capsid protein of HIV virus or from cholecystokinin-22. None of the reactions carried out yielded the desired condensation products. However, when Fmoc-NLQNLDPSHR-OH and cholecystokinin-12 (H-ISDRDYMGWMDF-NH2) were used as substrates, the last had its R-D peptide bond hydrolyzed producing cholecystokinin-8. The proteolytic activity of this enzyme measured against a fluorogenic peptide derivative was 50 times lower in DMF/TFE containing 5% of aqueous phase than in buffer. Steady-state fluorescence studies in DMF/TFE buffer were performed to examine the structure of this protease in these media. Steady-state spectra obtained with increasing proportions of these two organic solvents in buffer showed that the emission intensities built up. Quenching studies with iodide revealed that the Io/I ratio (where Io and I are the fluorescence emission intensities in the absence and presence of quencher, respectively) changed from 1.2 in aqueous media to 2.2 in DMF/TFE (1:1, v/v) containing 11% 0.2 M Tris-HCl buffer, pH 8.0, for 0.5 M iodide. The complete data indicated a higher exposure of tryptophan residues to the quencher in organic media, probably because of the partial unfolding of the enzyme.

Amino Acid Sequence↗