Mario Altamirano-Lozano

Total antioxidant levels, gender, and age as risk factors for DNA damage in lymphocytes of the elderly.

During past years, the association of oxidative stress with DNA damage and its possible clinical translation into chronic degenerative illnesses, such as atherosclerosis, cancer, diabetes mellitus and Alzheimers disease, has been demonstrated. In addition, it has been pointed out that age and gender are factors that influence the generation of DNA damage; however, this is still controversial. We have previously reported the results of a study of 88 subjects older than 60 years of age in whom DNA damage is related with serum levels of total antioxidants. The results of this study demonstrate a greater frequency of DNA damage in elderly persons with normal levels of antioxidants, in addition to males, and in the younger group of subjects, i.e., 60-69 years. In this work, we enlarged our study sample to 160 elderly subjects; in this way, we were able to evaluate the consistency of the influence of total antioxidants, age, and gender on the magnitude and grade of DNA damage in lymphocytes of the elderly. The results demonstrated that 45% of the subjects showed DNA damage, measured by an alkaline unicellular electrophoresis technique (comet assay). Similarly, 62% of the subjects presented low levels of total antioxidant levels measured by a colorimetric method (Randox Kit). A greater percentage of DNA damage was observed in subjects with normal levels of antioxidants (48%) compared with subjects with low levels (43%), although the difference was not statistically significant. The group of subjects 70 years of age or older showed a greater percentage of DNA damage (50%) than the group of subjects of 60-69 years of age (41%). However, the difference was again not statistically significant (P>0.05). With respect to gender, 64% of males and 38% of females had DNA damage with an odds ratio (OR) of 2.86 and a 95% confidence interval (CI) of 1.31-6.32 (P<0.05). In the logistic regression analysis, the interaction of the male sex variables with low antioxidants had an OR of 2.5 (CI 95%, 1.33-4.68; P<0.01). We conclude that the interaction of male sex factors-low levels of antioxidants would justify the indication of antioxidant dietetic supplements.

Reprotoxic and genotoxic studies of vanadium pentoxide in male mice

Effects of vanadium pentoxide (V2O5) treatment on reproductive function and testicular DNA in male mice were investigated. These functions were evaluated with fertility rate, implants, resorptions, sperm counts, motility, and morphology. The DNA damage in individual testis cells was analyzed by single-cell gel electrophoresis technique (COMET assay). V2O5 treatment resulted in a decrease in fertility rate, implantations, live fetuses, and fetal weight, and an increase in the number of resorptions/dam. Sperm count, motility, and morphology were impaired with the advancement of treatment. Vanadium treatment induced DNA damage depending on the dose in the testis cells that was expressed and detected as DNA migration in the COMET assay. The distribution of DNA migration among cells, a function of dose, revealed that the majority of cells of treated animals expressed more DNA damage than cells from control animals. It is concluded that vanadium pentoxide was a reprotoxic and genotoxic agent in mice.

Genotoxic effects of vanadium(IV) in human peripheral blood cells

Vanadium has been considered an aneuploidogen; however, there is controversial information about the clastogenic effects of vanadium compounds. In this study, the genotoxicity of vanadium(IV) tetraoxide (V(2)O(4)) was evaluated in human cultured lymphocytes and leukocytes using the mitotic index (MI), the replicative index (RI), chromosome aberrations (CA), sister chromatid exchanges (SCE), satellite associations (SA) and the single cell gel electrophoresis (SCGE) assay. This chemical induced a clear dose-response in MI inhibitions and modifications in the RI. In the CA, including breaks and exchanges and in the SCE, a significant increase appeared in the treated group compared with the controls. The SA test did not reveal an important difference. For the detection of genotoxic properties of vanadium(IV) using the SCGE assay, the 2 h evaluation period was not long enough for the chemical to enter the cell. These results indicate that vanadium(IV) tetraoxide is capable of inducing cytotoxic and cytostatic effects and chromosomal damage.

Genetic toxicology of thallium: a review.

This review summarizes the current knowledge about the general toxicity of thallium (Tl) and its environmental sources, with special emphasis placed on its potential mutagenic, genotoxic, and cytotoxic effects on both eukaryotic and prokaryotic cells. Tl is a nonessential heavy metal that poses environmental and occupational threats as well as therapeutic hazards because of its use in medicine. It is found in two oxidation states, thallous (Tl+) and thallic (Tl3+), both of which are considered highly toxic to human beings and domestic and wild organisms. Many Tl compounds are colorless, odorless and tasteless, and these characteristics, combined with the high toxicity of TI compounds, have led to their use as poisons. Because of its similarity to potassium ions (K+), plants and mammals readily absorb Tl+ through the skin and digestive and respiratory systems. In mammals, it can cross the placental, hematoencephalic, and gonadal barriers. Inside cells, Tl can accumulate and interfere with the metabolism of potassium and other metal cations, mimicking or inhibiting their action. The effects of Tl on genetic material have not yet been thoroughly explored, and few existing studies have focused exclusively on Tl+. Both in vivo and in vitro studies indicate that Tl compounds can have a weak mutagenic effect, but no definitive effect on the induction of primary DNA damage or chromosomal damage has been shown. These studies have demonstrated that Tl compounds are highly toxic and lead to changes in cell-cycle progression.

DNA damage induction in human cells exposed to vanadium oxides in vitro

Vanadium and vanadium salts cause genotoxicity and elicit variable biological effects depending on several factors. In the present study, we analyzed and compared the DNA damage and repair processes induced by vanadium in three oxidation states. We used human blood leukocytes in vitro and in a single cell gel electrophoresis assay at two pH values. We observed that vanadium(III) trioxide and vanadium(V) pentoxide produced DNA single-strand breaks at all of the concentrations (1, 2, 4, or 8 μg/ml) and treatment times (2, 4, or 6 h) tested. Vanadium(IV) tetraoxide treatment significantly increased DNA damage at all concentrations for 4 or 6 h of treatment but not for 2 h of treatment. The DNA repair kinetics indicated that most of the cells exposed to vanadium III and V for 4 h recovered within the repair incubation time of 90 min; however, those exposed to vanadium(IV) repaired their DNA within 120 min. The data at pH 9 indicated that vanadium(IV) tetraoxide induced DNA double-strand breaks. Our results show that the genotoxic effect of vanadium can be produced by any of its three oxidation states. However, vanadium(IV) induces double-strand breaks, and it is known that these lesions are linked with forming structural chromosomal aberrations.

Genotoxic studies of vanadium pentoxide (V2O5) in male mice. II. Effects in several mouse tissues

Vanadium pentoxide (V2O5) was tested for its ability to induce genotoxic damage in six different organs (liver, kidney, lung, spleen, heart, and bone marrow) of mice by using the alkaline Single Cell Gel Electrophoresis (SCGE) assay. Animals were sacrificed 24 h after i.p. administration of the vanadium pentoxide of 23.0, 11.5, or 5.75 microg/g (corresponding to the LD50, 1/2 LD50 and 1/4 LD50, respectively). In all tissues and organs evaluated (except for bone marrow), V2O5 increased the number of cells with damage. Our results showed that i.p. injection of V2O5 induced DNA damage in different organs and tissues, and that this kind of damage can be observed even 24 h after treatment. The analysis of DNA migration and the distribution of DNA damage showed that there are differences in sensitivity between organs and tissues to this compound. In addition the sensitivity of SCGE assay allows the detection of long term DNA damage and the possibility to compare it in various tissues and target organs.

Antigenotoxic Effects of (–)-Epigallocatechin-3-Gallate (EGCG), Quercetin, and Rutin on Chromium Trioxide-Induced Micronuclei in the Polychromatic Erythrocytes of Mouse Peripheral Blood

This study was conducted to investigate the modulating effects of (–)-epigallocatechin-3-gallate (EGCG), quercetin, and rutin on the genotoxic damage induced by Cr(VI) in polychromatic erythrocytes of CD-1 mice. The animals were divided into the following groups: (i) vehicle only; (ii) flavonoids (10 mg/kg EGCG, 100 mg/kg quercetin, 625 mg/kg rutin, or 100–625 mg/kg quercetin–rutin); (iii) Cr(VI) (20 mg/kg of CrO3); and (iv) flavonoids concomitantly with Cr(VI). All of the treatments were administered intraperitoneally (ip). The genotoxic damage was evaluated based on the number of micronucleated polychromatic erythrocytes (MN-PCE) obtained from the caudal vein 0, 24, 48, and 72 h after treatment. Groups treated with EGCG and quercetin exhibited no significant statistical changes in induction of MN-PCE. However, CrO3 treatment significantly increased MN-PCE induction 24 and 48 h after injection. Treatment with flavonoids prior to CrO3 exposure decreased MN-PCE induction compared with CrO3 only. The magnitudes of the potency of flavonoids were in the following order: rutin (82%) > quercetin (64%) > quercetin–rutin (59%) and EGCG (44%). The group treated with rutin significantly reduced genotoxic damage in mice treated with Cr(VI) (antioxidant effect). However rutin exerted a marginal genotoxic effect when administered alone (pro-oxidant effect). Our findings suggest protective effects of EGCG, quercetin, and rutin against genotoxic damage induced by Cr(VI).

Effect of chlorophyllin on chromium trioxide-induced micronuclei in polychromatic erythrocytes in mouse peripheral blood.

The effect of chlorophyllin on micronucleated polychromatic erythrocytes (MN-PCE) induction by chromium trioxide (CrO(3)) exposure in peripheral blood of mice was studied. Animals were treated with a single intraperitoneal dose of chlorophyllin (CHL) (20mg/kg), CrO(3) (20mg/kg), and CHL (20mg/kg) 4h before (CHL-CrO(3)) or 4h before and 20h after chromium treatments (20mg/kg; CHL-CrO(3)-CHL). Peripheral blood samples were drawn from the caudal vein at 0, 12 and 48h, and analyzed by the acridine orange (AO) technique. The results obtained in present study shown that CHL injection did not modify the number of MN-PCE. CrO(3) treatment resulted in a significantly increases 12 and 48h after the injection, reaching a four-fold increase 48h after CrO(3) administration. Whereas treatment with 20mg/kg of CHL prior to chromium, decreased the MN frequency induced by chromium in the 12h samples. When the samples were analyzed 48h after CrO(3) injection, no significant differences between CHL-CrO(3) and CHL-CrO(3)-CHL in comparison with CrO(3) treatment, were observed. These results indicate that increase of MN-PCE by CrO(3) is CHL-blocked in both protocols used (CHL-CrO(3) and CHL-CrO(3)-CHL) at 12h after treatment, but it was unable to modify the frequency of MN-PCE measured 48h after CrO(3) injection. The absence of a protective effect by CHL in the MN-PCE induction by CrO(3) at 48h, show that CHL has action only on one of the times of MN induction and suggests the possible action of CrO(3) by two different mechanisms, and not by CHL time-limited in vivo.

High Dosage of Ascorbic Acid and Alpha-Tocopherol Is Not Useful for Diminishing Oxidative Stress and DNA Damage in Healthy Elderly Adults

Aim: To determine the useful dosage of ascorbic acid and alpha-tocopherol against oxidative stress and DNA damage in the elderly. Methods: A double-blind controlled clinical assay carried out in a sample of 66 healthy subjects divided into three age-paired random groups with 22 subjects in each group. Group A received placebo and group B was administered 500 mg of ascorbic acid and 400 IU of alpha-tocopherol, whereas group C received 1,000 mg of ascorbic acid and 400 IU of alpha-tocopherol for a 6-month period. The following measurements were performed before and after the 6-month treatment period: thiobarbituric acid reactive substances (TBARS); total antioxidant status (TAS); superoxide dismutase (SOD), and glutation peroxidase (GPx) and DNA damage by comet assay. Results: After 6 months, group B subjects exhibited an increase in SOD and GPx enzyme levels; however, this was not statistically significant (p > 0.05). Likewise, TBARS and TAS concentrations remained unchanged (p > 0.05). In addition, in group C the decrease in TBARS and increase in SOD, GPx, and TAS were not statistically significant (p > 0.05). Similarly, average DNA migration showed no significant differences with high-dosage ascorbic acid and alpha-tocopherol. Conclusion: These findings suggest that administration of 1,000 mg of ascorbic acid plus 400 IU of alpha-tocopherol for 6 months is not useful for diminishing oxidative stress and DNA damage in healthy elderly adults.

DNA damage in lymphocytes of elderly patients in relation with total antioxidant levels.

DNA damage may occur as a result of an imbalance between the production and removal of free radicals, a process in which age plays an outstanding role. The purpose of this study was to analyze the relationship between total antioxidants and DNA damage in a sample of old age people in Mexico City. The sample included a total of 88 subjects, 15 males and 69 females, with a mean age of 65.5 years old (range between 60 and 79 years old), all of whom had lived in Mexico City during the last 10 years and had been diagnosed as clinically healthy. Results showed that 52% of the subjects presented DNA damage in peripheral blood lymphocytes which was assessed through an alkaline unicellular electrophoresis procedure (Comet Test), regardless of total antioxidant serum levels quantified through a colorimetric method (Randox Kit). Higher non-damage occurrences were observed in subjects with low antioxidant levels, a difference that was statistically significant (P < 0.05). Furthermore, the highest incidence of damaged cells was observed in subjects belonging to the 70-years-old-and-above group (P < 0.05). As to the magnitude and intensity of the damage associated to total antioxidant concentrations, a trend toward greater DNA damage in subjects with low serum levels was observed. It is concluded that low antioxidant levels are not always indicative of oxidative strain and therefore should not be considered as predictors of DNA damage in this population.