Preliminary safety assessment of C-8 xylitol monoester and xylitol phosphate esters.

OBJECTIVES: Most of the cosmetic compounds with preservative properties available in the market pose some risks concerning safety, such as the possibility of causing sensitization. Due to the fact that there are few options, the proper development of new molecules with this purpose is needed. Xylitol is a natural sugar, and the antimicrobial properties of xylitol-derived compounds have already been described in the literature. C-8 xylitol monoester and xylitol phosphate esters may be useful for the development of skincare products. As an initial screen for safety of chemicals, the combination of in silico methods and in vitro testing can aid in prioritizing resources in toxicological investigations while reducing the ethical and monetary costs that are related to animal and human testing. This study was designed to evaluate the safety of C-8 xylitol monoester and xylitol phosphate esters regarding carcinogenicity, mutagenicity, skin and eye irritation/corrosion and sensitization through alternative methods. METHODS: For the initial safety assessment, quantitative structure-activity relationship methodology was used. The prediction of the parameters carcinogenicity/mutagenicity, skin and eye irritation/corrosion and sensitization was generated from the chemical structure. The analysis also comprised physical-chemical properties, Cramer rules, threshold of toxicological concern and Michael reaction. In silico results of candidate molecules were compared to 19 compounds with preservative properties that are available in the market. Additionally, in vitro tests (Ames test for mutagenicity, cytotoxicity and phototoxicity tests and hen's egg test--chorioallantoic membrane for irritation) were performed to complement the evaluation. RESULTS: In silico evaluation of both molecules presented no structural alerts related to eye and skin irritation, corrosion and sensitization, but some alerts for micronucleus and carcinogenicity were detected. However, by comparison, C-8 xylitol monoester, xylitol phosphate esters showed similar or better results than the compounds available in the market. Concerning experimental data, phototoxicity and mutagenicity results were negative. As expected for compounds with preservative activity, xylitol-derived substances presented positive result in cytotoxicity test. In hen's egg test, both molecules were irritants. CONCLUSION: Our results suggested that xylitol-derived compounds appear to be suitable candidates for preservative systems in cosmetics.

ß-1,3-Glucan given orally modulates immunomyelopoietic activity and enhances the resistance of tumour-bearing mice.

ß-Glucans have been reported to be potent adjuvants in stimulating innate and adaptive immune responses. The aim of the present study was to determine the immunohematopoietic effects of Imunoglucan (HEBRON) following its oral administration to normal and Ehrlich ascites tumour (EAT)-bearing mice. Mice were treated with 250, 500 and 1000 mg/kg per day, p.o., Imunoglucan (ß-1,3-glucan extracted from Saccharomyces cerevisae) for 18 consecutive days. Treatment started 10 days prior to and ended 8 days after tumour inoculation. At 500 and 1000 mg/kg per day, Imunoglucan enhanced the life span of EAT-bearing mice and prevented myelosuppression and splenomegaly caused by the tumour by increasing the number of granulocyte-macrophage progenitors in the bone marrow and increasing colony-stimulating activity in the serum. At 500 mg/kg, Imunoglucan restored the reduced ability of stromal cells to display myeloid progenitors in long-term bone marrow cultures of EAT-bearing mice and upregulated the production of interleukin (IL)-6 and IL-1α by these cells, consistent with a higher number of non-adherent cells. Moreover, 500 mg/kg Imunoglucan restored natural killer cell activity in tumour-bearing mice, consistent with the increased production of interferon (IFN)-γ observed. The results of the present study suggest that Imunoglucan given orally indirectly modulates immune activity and probably disengages tumour-induced suppression by producing a higher reserve of myeloid progenitors in the bone marrow in consequence of biologically active cytokine release (colony-stimulating factors, IL-1α, IL-6 and IFN-γ).

Bcl-2 expression and apoptosis induction in human HL60 leukaemic cells treated with a novel organotellurium(IV) compound RT-04.

Organotellurium(IV) compounds have been reported to have multiple biological activities including cysteine protease-inhibitory activity, mainly cathepsin B. As cathepsin B is a highly predictive indicator for prognosis and diagnosis of cancer, a possible antitumor potential for these new compounds is expected. In this work, it was investigated the effectiveness of organotellurium(IV) RT-04 to produce lethal effects in the human promyelocytic leukaemia cell line HL60. Using the MTT tetrazolium reduction test, and trypan blue exclusion assay, the IC50 for the compound after 24 h incubation was 6.8 and 0.35 microM, respectively. Moreover, the compound was found to trigger apoptosis in HL60 cells, inducing DNA fragmentation and caspase-3, -6, and -9 activations. The apoptsosis-induced by RT-04 is probably related to the diminished Bcl-2 expression, observed by RT-PCR, in HL60-treated cells. In vivo studies demonstrated that the RT-04 treatment (2.76 mg/kg given for three consecutive days) produces no significant toxic effects for bone marrow and spleen CFU-GM. However, higher doses (5.0 and 10 mg/kg) produced a dose-dependent reduction in the number of CFU-GM of RT-04-treated mice. These results suggest that RT-04 is able to induce apoptosis in HL60 cells by Bcl-2 expression down-modulation. Further studies are necessary to better clarify the effects of this compound on bone marrow normal cells.

Phenotypic and genotypic characterisation of Escherichia coli strains serogrouped as enteropathogenic E. coli (EPEC) isolated from pasteurised milk.

Fifty-six Escherichia coli strains, serogrouped as EPEC, isolated from three different brands of pasteurised milk commercialised in Rio de Janeiro, Brazil, were tested for enteropathogenicity markers. Most of the strains (71.4%) were adherent to HEp-2 cells. The adherent strains were distributed among 7 EPEC serogroups (O26, O55, O111, O114, O125, O127, O128, O158). Although almost half of these strains (33.9%) presented unrecognisable adherence phenotypes, classical adherence patterns (localised-like, aggregative and diffuse adherence) described for E. coli and epidemiologically associated with diarrheagenic strains were observed. None of the strains showed typical localised adherence, usually associated with EPEC strains, but 4 of them displayed a localised-like adherence (LAL) phenotype, characterised by fewer and less compact microcolonies but that is still associated with diarrheagenic strains as well as strains of non-human origin. Indeed, 3 of these 4 strains were able to elicit the attaching-effacing lesion (FAS-positive), the central feature of EPEC pathogenesis, and hybridised with bfpA and eae DNA probes. The other LAL-positive strain hybridised with the bfpA probe but gave negative results for the eae probe and FAS assays. Interestingly, all LAL-positive strains produced amplicons of 200 bp in the PCR for bfpA, instead of the expected 326 bp fragment. PCR reactions for stx1 and stx2, two shiga-toxin-encoding genes, gave negative results. Typing of LEE-associated genes by PCR showed the profile eae (beta), tir (beta), espA (alpha) and espB (alpha) for one of the LAL-positive strain. The most prevalent adherence phenotype was the aggregative pattern which is observed in strains epidemiologically associated with persistent diarrhea. Additionally, one strain promoted complete detachment of the Hep-2 cell monolayer after 3 h of infection which might be related to the production of citotoxins, a feature that has been increasingly observed in clinical strains. The possession of EPEC-related O and H antigens is no longer deemed an essential characteristic of true pathogenic EPEC strains, emphasising the importance of routinely screen for virulence markers in E. coli strains isolated from foods. Our results are in accordance with data from the literature that demonstrate that environmental strains display atypical features but yet are capable of eliciting the classical A/E lesion and thus must be considered as potentially pathogenic. Further, our results demonstrate the potential of pasteurised milk as a vehicle for transmission of diarrheagenic E. coli in Brazil.

Myelopoietic response in tumour-bearing mice by an aggregated polymer isolated from Aspergillus oryzae.

The effects of magnesium ammonium phospholinoleate-palmitoleate anhydride (MAPA), a proteic aggregated polymer isolated from Aspergillus oryzae, on the growth and differentiation of granulocyte-macrophage progenitor cells (colony-forming unit-granulocyte-macrophage [CFU-GM]) in normal and Ehrlich ascites tumour-bearing mice were studied. Myelosuppression concomitant with increased numbers of spleen CFU-GM was observed in tumour-bearing mice. Treatment of these animals with MAPA (0.5-10 mg/kg) stimulated marrow myelopoiesis in a dose-dependent manner and reduced spleen colony formation. No changes were observed in total and differential marrow cell counts. The dose of 5.0 mg/kg MAPA, given prior or after tumour inoculation, was the optimal biologically active dose in tumour-bearing mice and this dose schedule also stimulated myelopoiesis in normal mice. MAPA significantly enhanced survival and concurrently reduced tumour growth in the peritoneal cavity. We propose that the modulatory effect of MAPA on the myelopoietic response may be related to its antitumour activity as a possible mechanism for regulation of granulocyte-macrophage production and expression of functional activities.

Effects of deltamethrin on the growth and differentiation of bone marrow hematopoietic stem cells.

1. The effects of deltamethrin on mouse bone marrow and spleen progenitor cell responsiveness to granulocyte and macrophage colony-stimulating factors (CSFs) were evaluated. 2. Deltamethrin (1-5 mg/kg) was administered four times subcutaneously on alternate days for one week to male BALB/c mice, 5-8 weeks old (N = 6 mice/group), raised under pathogen-free conditions and maintained in conventional animal rooms for four weeks before use. Soft agar colony formation (CFU-C), marrow and spleen cell counts as well as body, spleen and thymus weights were determined. 3. Although treatment with the lowest dose (1 mg kg-1 48 h-1) produced no significant effect on CFU-C, the administration of 3 and 5 mg kg-1 48 h-1 caused a more than two-fold increase in the formation of granulocyte and macrophage colonies in the marrow, but not in the spleen (control value = 100.5 +/- 12 for N = 6). Colony numbers returned to normal values within five days after the end of deltamethrin administration. 4. No changes were observed in the total (range: 1-3 x 10(8) per spleen) and differential marrow and spleen cell counts, nor was there any alteration in spleen weight. However, treatment with the three doses resulted in a dramatic reduction in thymus weight. 5. These effects were not due to the liberation of endotoxin, because if endotoxin had been present it would have been < 0.060 ng/ml, a concentration that would not have a biological effect. 6. In vitro addition of 0.10 to 10 microM deltamethrin to marrow cell cultures obtained from untreated mice did not induce any response. 7. These data indicate that the CSF-driven granulocyte and macrophage development provides a useful model for the study of the effects of toxicants on myelopoiesis.

Measurement of the respiratory burst and chemotaxis in polymorphonuclear leukocytes from mercury-exposed workers.

The chemotactic and nitroblue tetrazolium reducing activities of neutrophils from 48 mercury-exposed workers were examined and compared with those of non-exposed, age- and sex-matched individuals. At the time of testing, the exposed population had a mean (+/- s.d.) urinary mercury concentration of 24.0 +/- 20.1 micrograms g-1 creatinine and in 44 of these workers urinary mercury levels were below the accepted threshold level (TLV) of 50 micrograms g-1 creatinine. The two neutrophil functions were significantly reduced in the mercury-exposed workers compared with the controls. In 28 of these workers, chemotaxis was re-evaluated 6 months later. During the intervening 6 months, the level of hygiene was improved throughout the plant and urinary mercury concentrations were determined monthly in each worker. Despite a significant reduction in urinary mercury concentrations, neutrophil migration did not return to within the normal range. These results suggest that 'safe' level mercury exposure may lead to impairment of neutrophil function.

Stimulation of myelopoiesis in Listeria monocytogenes-infected mice by an aggregated polymer isolated from Aspergillus oryzae.

In this work, we investigated the effects of the proteic aggregated polymer of magnesium ammonium phospholinoleate-palmitoleate anhydride (MAPA) isolated from Aspergillus oryzae on the growth and differentiation of bone marrow granulocyte-macrophage progenitor cells (CFU-GM) in Listeriamonocytogenes-infected mice. A significant reduction in the CFU-GM number was observed in the initial phase of infection with a sublethal dose of Listeria. Treatment of mice with 0.5, 2.0 and 5.0 mg/kg MAPA for 7 days prior to infection significantly stimulated myelopoiesis in a dose-dependent manner. Moreover, treatment with 0.5 and 5.0 mg/kg MAPA resulted in 30% and 40% cures of mice lethally infected with Listeria, respectively. MAPA added directly to the culture dishes hardly affected colony formation by bone marrow cells, suggesting an indirect effect ofthis compound on myelopoiesis in vivo. In summary, the data show that MAPA can modulate the CFU-GM generation and antibacterial resistance in listeriosis. As the ability of hematopoietic tissues to produce phagocytes is of particular significance to mediate resistance to Listeria, the promotion of bone marrow CFU-GM by MAPA may contribute to a rapid restoration of phagocyte numbers in infected sites, thus mitigating the course of infection.

Defective neutrophil function in workers occupationally exposed to hexachlorobenzene.

In this work we have studied the respiratory burst and chemotaxis of polymorphonuclear leukocytes from 51 workers exposed to chlorinated compounds, which were compared with those of non-exposed, age- and sex-matched individuals. These two neutrophil functions were significantly reduced as compared to controls. No correlation was observed between the length of exposure, hexachlorobenzene (HCB) blood concentrations and neutrophil chemotaxis or the extent of nitroblue tetrazolium reduction.

Abnormal antioxidant system in erythrocytes of mercury-exposed workers.

To investigate the effects of chronic exposure to mercury we studied the red cell antioxidant system in mercury-exposed workers through the evaluation of reduced glutathione, catalase and superoxide dismutase systems. Of these workers, some were being exposed at the time and had presented urinary mercury levels considered safe for occupational exposure for at least 3 months prior to the initiation of this study, and others had been on leave for at least 6 months because of intoxication symptoms. Reduced glutathione levels were lower and catalase activity was higher in the workers which were still being exposed, compared to those on leave and controls. No differences were observed between the workers on leave and controls.

Immunoglobulin levels in workers exposed to hexachlorobenzene.

The serum immunoglobulin (IgG, IgM and IgA) concentrations of 52 chlorinated-exposed workers were examined and compared with those of non-exposed, age- and sex-matched individuals. At the time of testing, the exposed population had mean hexachlorobenzene (HCB) blood levels of 3.84 micrograms/dl with a range of 0.1 to 16 micrograms/dl. Increased IgG and IgM levels were found in the HCB-exposed workers (P < 0.05 and P < 0.01, respectively). Hepatic function was evaluated by serum aspartate (AST) and alanine aminotransferase (ALT) activities, as well as by bilirubin levels. IgM concentrations were positively correlated with three of the studied parameters, namely, length of exposure (r = 0.367) and the activities of both AST (r = 0.367) and ALT (r = 0.507).

Active immunization against hepatitis B virus (HBV) with low-doses of plasma-derived vaccine by intradermal route.

Schedule for vaccination against HBV infection has usually been based on three separate injections of 20 mcg of the vaccine by intramuscular route. One of the main shortcomings to its use in large scale programs has been its high cost. Ninety out of 300 health workers were submitted to three injections of 2 mcg of plasma-derived vaccine (PDV) by intradermal (ID) route on days 0, 30, and 180. Anti-HBs was detected in 74 (82.2%) after the second dose and in 80 (88.9%) after the third dose, a non-significant difference. However, levels above 10 times the cut-off were observed in 29 (32.2%) and 77 (85.5%), respectively (p less than 0.001). The results showed that a low-dose schedule is effective when used in health workers and should be tried with other risk groups.

Isolation and serological identification of enteropathogenic Escherichia coli in pasteurized milk in Brazil.

OBJECTIVE: To evaluate the microbiological quality of pasteurized milk commercialized in Rio de Janeiro, Brazil, and determine serologically enteropathogenic Escherichia coli (EPEC) strains in E. coli isolates obtained from milk samples. METHODS: Ninety samples of pasteurized milk - types B and C - of three different commercial brands, purchased in supermarkets and bakeries in Rio de Janeiro, were examined. The amount of total and fecal coliform bacteria was estimated using the Most Probable Number technique. Mesophilic, psychrotrophic, and thermoduric microorganism counts were determined by the Standard Plate Count technique. Isolation and identification of E. coli were carried out using conventional physiological tests. Commercial antisera were used for serological characterization of EPEC. RESULTS: The three milk brands analyzed revealed bacterial counts above the regulated values of the Brazilian government. It was found that among 208 strains of E. coli isolated, 46 (22.1%) were serologically classified as EPEC. The most common EPEC serogroup was O55 (15.2%). CONCLUSIONS: Though recent studies on virulence factors indicate that not all strains serologically classified as EPEC are able to attaching/effacing lesion, it is believed that the isolation of EPEC serogroups from pasteurized milk represent a potential risk for children, as well as an indicative of the presence of other enteropathogens.

3,4-methylenedioxymethamphetamine (MDMA--Ecstasy) decreases neutrophil activity through the glucocorticoid pathway and impairs host resistance to Listeria monocytogenes infection in mice.

Ecstasy is the popular name of the abuse drug 3,4-methylenedioxymethamphetamine (MDMA) that decreases immunity in animals. The mechanisms that generate such alterations are still controversial. Seven independent pharmacological approaches were performed in mice to identify the possible mechanisms underlying the decrease of neutrophil activity induced by MDMA and the possible effects of MDMA on host resistance to Listeria monocytogenes. Our data showed that MDMA (10 mg kg(-1)) administration decreases NFκB expression in circulating neutrophils. Metyrapone or RU-486 administration prior to MDMA treatment abrogated MDMA effects on neutrophil activity and NFκB expression, while 6-OHDA or ICI-118,551 administration did not. As MDMA treatment increased the plasmatic levels of adrenaline and noradrenaline, propranolol pre-treatment effects were also evaluated. Propranolol suppressed both MDMA-induced increase in corticosterone serum levels and its effects on neutrophil activity. In a L. monocytogenes experimental infection context, we showed that MDMA: induced myelosuppression by decreasing granulocyte-macrophage hematopoietic progenitors (CFU-GM) in the bone marrow but increased CFU-GM in the spleen; decreased circulating leukocytes and bone marrow cellularity and increased spleen cellularity; decreased pro-inflammatory cytokine (IL-12p70, TNF, IFN-γ, IL-6) and chemokine (MCP-1) production 24 h after the infection; increased the production of pro-inflammatory cytokines and chemokines 72 h after infection and decreased IL-10 levels at all time points analyzed. It was proposed that MDMA immunosuppressive effects on neutrophil activity and host resistance to L monocytogenes rely on NFκB signaling, being mediated by HPA axis activity and corticosterone.

Interactions of clinical and environmental Aeromonas isolates with Caco-2 and HT29 intestinal epithelial cells.

AIM: Evaluation of adherence and invasion of Aeromonas spp. to human colon carcinoma cell lines Caco-2 and HT29 and assessment of cytotoxic activity. METHODS AND RESULTS: A number of 27 strains of Aeromonas caviae and 23 strains of Aeromonas hydrophila was analysed. All strains were capable to adhere to sub-confluent monolayers of Caco-2 and HT29 cell types, presenting aggregative and diffuse adherence patterns cells, respectively. In the cytotoxic assays all strains showed cytopathic and/or cytotoxic activities to Vero cells. The evaluation of the tetrazolium salt (MTT test) reduction capability was carried out in Vero, Caco-2, and HT29 cells. MTT test showed that Vero cell line was the most sensitive cell type. In the invasion test, 13 strains were analysed on Caco-2 and HT29 monolayers. Only two (15%) of the 13 strains, A. hydrophila and A. caviae species, both isolated from vegetables were invasive to Caco-2 cells. No strains were able to invade the HT29 cells. CONCLUSIONS: A. hydrophila and A. caviae isolated from human diarrhoeic faeces, vegetables, and water, were able to adhere to and produce cytotoxic/cytopathic effects in intestinal epithelial cell lines. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of Aeromonas spp. in food and water samples expressing virulence factors suggest that these sources may act as dissemination vehicles of human pathogen with implication in the public health.

Early proliferation of umbilical cord blood cells from premature neonates.

BACKGROUND AND OBJECTIVE: Human umbilical cord blood (UCB) is an important source of haematopoietic stem cells; however, the behaviour of progenitor cells obtained from premature and full-term neonates is still a controversy subject. Thus, the aim of this study was to evaluate cell cycle parameters and the proliferative capacity of UCB progenitor cells from premature and full-term neonates. MATERIAL AND METHODS: Clonogenic assays were performed with methylcellulose, medium supplemented with recombinant stimulating growth factors and the colonies were scored on the seventh day and the 14th day of culture. A cell cycle study was carried out by DNA analysis using flow cytometry and 30 000 events were acquired; p107 and p130 expressions were analysed by Western blotting. RESULTS: Cultures obtained from UCB of premature neonates showed an early growth of colony-forming unit (CFU)-burst forming unit erythroid/CFU-granulocyte, erythrocyte, macrophage and megakaryocyte (BFU-E/GEMM), and CFU-granulocyte, macrophage (GM) by the seventh day of culture (P < 0.001). Therefore, the number and morphological characteristics of these colonies were comparable with those obtained from full-term neonates, on the 14th day of culture. At the 14th day, a large amount of CFU-GM was detected in the premature group (P < 0.0032). The premature culture on the 14th day showed fibroblasts and was comparable to those of full-term neonates on the 21st day in terms of number and morphology of the colonies. DNA analysis showed that the number of cells in S-phase was also higher in premature samples when compared to full-term neonates, P < 0.0021 (0 h = 12.8 vs. 2.5%; 16 h = 10.5 vs. 5.9%; 20 h = 13.5 vs. 10.3%; 24 h = 13.8 vs. 9.1%; 48 h = 14.0 vs. 5.4%; 72 h = 20.5 vs. 8.9%; 96 h = 13.8 vs. 7.7%). The Western blotting results demonstrated that p107 and p130 cell cycle protein expressions were higher in premature cells than in full-term cells. CONCLUSION: These results suggest that the higher capacity of proliferation and early differentiation of premature UCB might not be related only to the amount of stem/progenitor cells but also to a different timing of cell cycle entry.

Hematopoietic effects in mice exposed to deltamethrin and hydrocortisone.

The effects of deltamethrin on bone marrow and spleen progenitor cell responsiveness to granulocyte and macrophage colony-stimulating factors were evaluated. Deltamethrin was tested in parallel with hydrocortisone to further investigate some similarity in the in vivo effects of both compounds observed in previous studies in our laboratory. In vivo effects were studied after the subcutaneous administration in mice of three 5 mg/kg injections of deltamethrin and a single 30 mg/kg injection of hydrocortisone to Balb/c mice. Soft agar colony formation, marrow and spleen cell counts, body, spleen and thymus weights were determined. Data obtained in vivo indicate that deltamethrin and hydrocortisone reversibly increase the formation of granulocyte and macrophage colonies in the marrow, but not in the spleen. No changes were observed in total and differential cell counts in the marrow and spleen and spleen weights. Treatment with both compounds, however, resulted in a dramatic reduction in thymus weights. Assays for endotoxin demonstrate that these effects were not due to the liberation of endotoxin. In vitro addition of 10(-5), 10(-6) and 10(-7) M deltamethrin and hydrocortisone to marrow cultures from untreated mice resulted in different effects from those observed in vivo. Hydrocortisone increased granulocyte and reduced macrophage colonies, whereas deltamethrin was without in vitro effects. It is suggested that deltamethrin effects are due to an indirect action on the hypothalamic-pituitary axis leading to increased corticosteroid levels. The importance of biotransformation mechanisms is also emphasized.

T lymphocytes in mercury-exposed workers.

In this work we have investigated the changes in T-helper and T-suppressor cells and T-cell proliferative response to phytohemagglutinin (PHA) in mercury-exposed workers. The study group consisted of 33 workers from a mercury-producing plant with a mean age of 29 years and a mean exposure period of 19 months. At the time of testing, and for the three previous months, the exposed population had urinary mercury levels below the currently accepted limit of 50 micrograms/g creatinine. A reverse CD4+/CD8+ ratio was observed in the mercury-exposed individuals which was characterized by a reduction in the number of CD4+ lymphocytes. No changes were observed in the proliferative response of lymphocytes from exposed individuals to PHA. Similarly, no proliferative response was observed when lymphocytes from normal individuals were cultivated in the presence of serum from the exposed workers. We found no correlations between lymphocytes changes and urinary mercury concentrations, time of exposure or the age of the workers.

Immunoglobulin E and autoantibodies in mercury-exposed workers.

In this work we have studied the serum immunoglobulin E (IgE) levels and the concentration of anti-DNA and anti-nucleus antibodies in mercury-exposed workers. The study group consisted of 36 workers from a mercury producing plant, with a mean age of 27 years and a mean exposure period to mercury of 19 months. At the time of testing, and for the three previous months, the exposed population had urinary mercury levels below the currently accept limit of 50 ug/g creatinine. Significantly increased IgE levels was found in the mercury-exposed individuals. Moreover, a moderate negative correlation (r = -0.43 P < 0.05) between the length of exposure to mercury and IgE levels was observed. Anti-DNA and anti-nucleus antibodies were not detected in these workers. These results suggest that the humoral immune response is an indicator of cellular changes in workers chronically exposed to mercury, even in those with urinary mercury concentrations within levels considered safe in the occupational area.