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Biomedical subjects

R Marcos

Publications and source records attributed to R Marcos.

At least 73 records · Page 4Linked to original sources
PubMed1998

Genotoxic activity of four inhibitors of DNA topoisomerases in larval cells of Drosophila melanogaster as measured in the wing spot assay.

Four inhibitors of DNA topoisomerases namely nalidixic acid, camptothecin, m-amsacrine and etoposide, have been evaluated for genotoxic effects in the wing spot test of Drosophila melanogaster. This assay assesses somatic recombination and mutational events. We studied nalidixic acid as an inhibitor of bacterial DNA gyrase, camptothecin as a topoisomerase I inhibitor, as well as m-amsacrine and etoposide as topoisomerse II inhibitors. The genotoxic effects were determined from the appearance of wing spots in flies trans-heterozygous for the recessive markers multiple wing hairs (mwh) and flare, as well as in flies heterozygous for mwh and the multiply inverted TM3 balancer chromosome. From our results it appears that whilst nalidixic acid and m-amsacrine were compounds that did not increase the incidence of mutant clones, camptothecin and etoposide proved to be significantly genotoxic in this test, being camptothecin more effective than etoposide. A significant proportion of the total spot induction was due to mitotic recombination, confirming previously reported data. On the other hand, the cotreatments of each topoisomerase inhibitor with the alkylating agent ethyl methanesulfonate (EMS) indicate that, while nalidixic acid, m-amsacrine and etoposide show a tendency to an antagonistic interaction, camptothecin shows an additive effect, suggesting mechanistic differences between the activity of the four inhibitors of DNA topoisomerases studied.

Amsacrine↗
PubMed1998

Granulomatous epididymo-orchitis: sonographic features and clinical outcome in brucellosis, tuberculosis and idiopathic granulomatous epididymo-orchitis.

PURPOSE: We reviewed and analyzed sonographic findings in granulomatous epididymo-orchitis in an attempt to differentiate this rare inflammatory entity from other causes of enlarged scrotum, such as tumors. MATERIALS AND METHODS: We retrospectively reviewed sonographic features of 9 patients a mean of 41 years old with specific and nonspecific granulomatous epididymo-orchitis, including brucellosis in 7, tuberculosis in 1 and idiopathic granulomatous epididymo-orchitis in 1. Histological confirmation was obtained in 3 cases, and in the remaining 6 positive clinical manifestations and laboratory tests for brucellosis were accepted as the main criteria for the diagnosis of brucellar epididymo-orchitis. RESULTS: The most notable echographic finding was an enlarged and heterogeneous epididymis, predominantly in the body and tail. Testicular involvement consisted of a diffusely hypoechoic testis or focal intratesticular areas. Thickening of the scrotal wall and tunica albuginea, and moderate hydrocele were also noted occasionally. Followup scans revealed intratesticular abscesses in 3 patients. CONCLUSIONS: These echographic findings may suggest the diagnosis of granulomatous epididymo-orchitis in the appropriate clinical setting, and help to avoid unnecessary orchiectomy for benign disease.

Adult↗
PubMed1998

Application of the single cell gel electrophoresis (SCGE) assay to the detection of DNA damage induced by 131I treatment in hyperthyroidism patients.

To provide further data on the possible applications of the single cell gel electrophoresis (SCGE) or Comet assay in human biomonitoring studies, we have evaluated the eventual genetic damage induced by therapeutic exposure to 131I, by measuring the tail length of the comet and the amount of DNA damage in peripheral blood cells from 16 hyperthyroidism patients treated with [131I]sodium iodide by oral administration. Blood samples were taken just before the treatment and 1 week and 1 month after. The results show a slight but significant increase in the mean tail length in the sample obtained 1 month after treatment. When the cells were classified according to the grade of damage the two post-treatment samples showed a clear increase in the proportion of damaged cells. The results of this study indicate that the DNA damage caused by 131I can be detected with the Comet assay, but when comparing the data reported here with our previous results obtained from the same patients and sampling times with the sensitive and well-established micronucleus test, the response in the Comet assay was less clear.

Adult↗
PubMed1998

Induced somatic and germinal reversion of the white-spotted-1 insertional mutant phenotype in Drosophila melanogaster.

The white-spotted-1 (Wsp1) mutant of Drosophila melanogaster is characterized by the presence of an 8.7 kb retrotransposon (B104) inserted in the regulatory region of the white locus. The frequency of reversion in both somatic tissue and the germline after exposure to three different alkylating agents has been analysed. To determine if germinal revertants were induced by precise excision of the insertional element we analysed several phenotypic revertants using PCR and Southern blot techniques. The results indicate that, under our experimental conditions, the mutagens used did not induce excision of B104 in the white gene. In addition, the revertant phenotypes obtained were due to the existence of second site modifiers acting on expression of white. Such modifiers map near the white locus and, at least in one case, may correspond to suppressor-of-white-spotted.

ATP-Binding Cassette Transporters↗
PubMed1998

Micronuclei analysis in lymphocytes of pesticide sprayers from Concepción, Chile.

To estimate the genetic risk associated with pesticide exposure in a defined population, the frequency of micronuclei (MN) in peripheral blood lymphocytes from a group of 22 pesticide sprayers from Concepción, Chile, occupationally exposed to pesticide mixtures was evaluated. After scoring 1,000 binucleated cells for each donor, no significant increases were observed either for the total number of MN or for binucleated cells with MN, when compared with a concurrent control population. In addition, when the effects of different confounding factors such as age, smoking, and drinking habits were considered, no significant effect was observed. Our conclusion is that, in this specific group of workers and under the particular conditions of exposure to pesticides, when evaluated by the micronucleus assay, no genetic risk was detected.

Adult↗
PubMed1997

Micronuclei, centromere-positive micronuclei and chromosome nondisjunction in cytokinesis blocked human lymphocytes following mitomycin C or vincristine treatment.

The influence of sampling time on the frequencies of micronuclei, centromere-positive micronuclei and chromosome nondisjunction was investigated in binucleated lymphocytes following treatment with a known clastogen (mitomycin C) or an aneuploidy-inducing agent (vincristine sulfate). Cytochalasin B (6 micrograms/ml) was added 44 h after mitogen stimulation and cultures were harvested 12, 28, 36 and 48 h thereafter. Micronucleated cells and micronuclei were significantly induced by the two treatments at all sampling times. Furthermore, in situ hybridization with an 'all centromeres' probe showed that vincristine-induced micronuclei were prevalently centromere-positive whereas in mitomycin C-treated cultures only a minor fraction of induced micronuclei contained the hybridization signals. Chromosome nondisjunction rates, as measured by in situ hybridization with chromosome 7- and 11-specific alphoid probes, significantly increased following vincristine treatment. Chromosome nondisjunction and total micronucleus frequencies were found to increase with time both in controls and in mutagen-treated cultures, whereas the percentage of centromere-positive micronuclei in the different treatments was not influenced by the sampling time. Our data suggest that even in the presence of 6 micrograms/ml cytochalasin B, the abnormal segregation of binucleated cells may contribute to the baseline level of micronuclei and influence the results obtained. The introduction of a short cytochalasin B treatment (between 12 and 28 h) in the cytokinesis-blocked micronucleus assay may avoid the cytochalasin B effect on micronucleus frequencies.

Aneuploidy↗
PubMed1997

SCE analysis in peripheral blood lymphocytes of a group of filling station attendants.

Petroleum derivatives constitute a complex mixture of chemicals which contain known genotoxicants. Thus, chronic occupational exposure to such derivatives may be considered to possess genotoxic risk. Service station attendants are workers exposed to petroleum derivatives and, as consequence, biomonitoring studies designed to evaluate the genetic risk are required. Here we present the data on sister chromatid exchange (SCE) frequencies, proliferation rate index (PRI) and cells with high frequency of SCE (HFC) from a group of 42 attendant workers. In addition, data from a group of 45 unexposed (controls) are presented. The results obtained indicate that there are no significant differences in SCE values between the exposed workers when compared to the non-exposed individuals. The only difference is found when smoking habit is considered, the smokers having significant increases in the SCE frequency. There is a slight increase in the HFC frequency of the occupationally exposed group but the difference did not attain statistical significance.

Adult↗
PubMed1997

Micronuclei induction by 131I exposure: study in hyperthyroidism patients.

To evaluate the eventual genetic damage induced by therapeutic exposure to 131I, we have studied the presence of micronuclei (MN) in binucleated peripheral blood lymphocytes from a group of 28 hyperthyroidism patients who received 131I sodium iodide, via oral administration. The study was conducted over time and blood samples were obtained before the treatment, and 1 week, 1 month and 3 months after it. The results obtained indicate a positive relationship between dose and BNMN frequency as calculated by the linear regression coefficient, showing significant increases in the frequency of MN and BNMN (binucleated cells with MN) in the subgroup of patients that received more than 500 MBq. Taking into account that the patients studied were treated with relatively low doses of 131I, our positive results support the view that the MN assay is sensitive enough to monitor the chromosome damage resulting from the exposure.

Administration, Oral↗
PubMed1997

Genotoxicity of humic acid in cultured human lymphocytes and its interaction with the herbicides alachlor and maleic hydrazide.

The genotoxicity of humic acid and its possible interaction with the herbicides alachlor and maleic hydrazide have been evaluated in cultured human lymphocytes from two donors. Humic acid and the two herbicides have been tested (alone and combined) for sister-chromatid exchange (SCE) induction. In addition, the effect of two different preincubation times, 2 and 24 hr, was analyzed. The results indicate that humic acid and the herbicides alachlor and maleic hydrazide appear to significantly enhance the frequency of SCE, the effect of the herbicides being more pronounced. With reference to the possible interaction of humic acid with the herbicides, the results do not show a common pattern, although mainly an additive effect was obtained. Nevertheless, there is some evidence suggesting that antagonism may occur, especially in the combined treatment of humic acid and maleic hydrazide.

Acetamides↗
PubMed1997

Decrease in platelet, erythrocyte and lymphocyte acetylcholinesterase activities due to the presence of protease inhibitors in the storage buffer.

The effect of the presence of protease inhibitors in the storage buffer on acetylcholinesterase (AChE) activity was studied in crude membrane preparations from sheep platelets before and after solubilization of the membranes with Triton X-100. Although sensitive to the action of trypsin, the biological activity of AChE remained unchanged for as long as 6 days at 4 degrees C in a protease-inhibitor-free medium. At 10(-5) M final concentration PMSF reduced AChE activity to 50% after 24 hours of storage. This reduction was abolished in mixtures in which PMSF was present together with 3 mM EGTA, 5 mM EDTA and 0.1 mg/mL trypsin inhibitor. Nevertheless, under these conditions, storage periods longer than 24 hours still drastically reduced AChE activity. A mixture of EDTA, EGTA and trypsin inhibitor also produced a decrease in AChE activity after 24 hours. A more complex cocktail of inhibitors including several commonly used peptides decreased AChE activity only if PMSF or EDTA were present in the mixtures. Similar results were obtained with sheep erythrocytes or lymphocytes, and bovine erythrocytes.

Acetylcholinesterase↗
PubMed1997

Aneugenic activity in human cultured lymphocytes. An overall study with colchicine using the micronucleus assay and fluorescence in situ hybridization techniques.

The effects induced by aneugenic agents on chromosome segregation are manifold. The biological relevance of these effects has led to the development of assays specifically detecting aneugens. In this context, the micronucleus (MN) assay in binucleated human lymphocytes along with FISH has been considered a pertinent tool for detecting aneugenic and clastogenic activity. However, the MN assay is insensitive in detecting aneugenic effects other than chromosome loss. By using the aneugenic model compound colchicine and X chromosome centromere-specific FISH, we have shown that besides chromosome loss in binucleated cells, other effects such as MN in mononucleated cells, cells arrested at metaphase, polyploidy and non-disjunction are also consistently induced by aneugenic agents. A chromosome 1 centromeric probe was used simultaneously with X chromosome centromeric labeling in mononucleated cells in order to distinguish polysomy from polyploidy. It is concluded that all these effects should be considered for a comprehensive evaluation of aneugenic activity.

Adult↗
PubMed1997

Radioactive iodine induces clastogenic and age-dependent aneugenic effects in lymphocytes of thyroid cancer patients as revealed by interphase FISH.

After the Chernobyl nuclear accident, a dramatic 131I-related increase in the incidence of thyroid cancer has been reported in exposed children. However, little is known about the eventual genotoxic effects of 131I in exposed humans. Thyroid cancer patients are usually treated with 131I and, therefore, they provide us with an opportunity to study cytogenetic damage induced by known doses of this radionuclide. FISH techniques have been employed to study the origin of micronuclei as well as X chromosome non-disjunction and X chromosome numerical abnormalities in lymphocytes from 131I-treated women suffering from thyroid cancer. Blood was sampled before and 1 week after 131I treatment. Cells were analysed with either pancentromeric FISH to classify micronuclei or X chromosome centromere-specific FISH in mononucleated and binucleated cells to evaluate X chromosome numerical abnormalities and non-disjunction respectively. Our data indicate that 131I-induced clastogenic and age-dependent aneugenic effects in the lymphocytes of exposed patients. The X chromosome was not preferentially involved in the aneugenic effect induced by 131I. It is concluded that besides its major clastogenic effect, 131I can also induce an X chromosome-independent aneugenic activity mainly in patients with spontaneous proneness to chromosome loss.

Adolescent↗
PubMed1997

Genotoxicity studies on ebrotidine.

Five genotoxicity studies on ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene)amino]-4-thiazolyl]methyl]thio] ethyl]amino]methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542), including at least four of the battery of tests recommended by toxicological regulatory guidelines for new drugs, were conducted. These tests were the Ames test for determination of bacterial gene mutations, sex-linked recessive lethal mutation test in Drosophila for gene mutations in eukaryotic systems, in vitro chromosome aberration test and micronucleus test for evaluation of structural and numerical aberrations, and sister chromatid exchange frequency test for assessment of non-specific damage to chromatin. Negative and positive controls were used in all the experiments. The effects were investigated in the absence or presence of metabolic activation by S-9 microsomal fraction from rat liver homogenate. A dose range toxicity study was also performed to determine the dosage levels or concentrations to be tested for the assessment of genotoxic effects. None of the tests showed a significant increase in the genotoxic parameters, both in vitro and in vivo in somatic or germ cells. It is, therefore, concluded that ebrotidine has not caused mutagenic or clastogenic effects in any of the experimental systems tested.

Animals↗
PubMed1997

Effects in the Comet assay of storage conditions on human blood.

The Comet assay is a rapid and sensitive method for analyzing single cells for DNA damage. Using human lymphocytes, the assay is particularly useful for human monitoring studies, as well as for in vitro genotoxicity testing of chemicals. In such studies, it is not always possible to collect and process matched samples on the same day as the blood is taken. It would be useful if some samples could be stored and examined at a different time, without loss of viability or other factors affecting responses. It is thus important to understand the effects of storage conditions on blood to be used in such studies and how exposure or treatment might modify such responses. In a joint study in two laboratories, blood was taken from various donors and stored under different conditions. It was examined on day 1 (day on which sample was taken) and days 2, 3, 4, 5, or 8 at room temperature, 4 degrees C, or -20 degrees C. Cells were treated after storage (from day 2 onward) with bleomycin (BLM) and ethylnitrosourea (ENU). The data were analyzed either by eye (classifying cells with different categories of damage) or by using a computerized image analysis system (Kinetic Imaging Ltd., Liverpool UK. Software Package Comet 3.0) where the tail moment, which is considered to be a sensitive measurement, has been analyzed. There was no loss of cell viability at 4 degrees C or room temperature up to 8 days when measured by trypan blue dye exclusion. Findings suggest that on days 1-4 for the untreated samples at room temperature or 4 degrees C there were no biologically meaningful changes in both the different categories of cell damage and tail moment data. In treated cultures up to day 4, either at room temperature or at 4 degrees C, responses were only minimally affected and changes were considered not to be of biological significance. However, there was slightly less variability between samples at 4 degrees C than at room temperature in one laboratory. The reverse was true in the other. This would suggest that samples can probably be stored up to day 4 at 4 degrees C or room temperature without any untoward effects. Provided samples can be processed within this 4-day time frame, it would not seem necessary to cryopreserve samples at -196 degrees C.

Bleomycin↗
PubMed1996

Genotoxic evaluation of the herbicide trifluralin on human lymphocytes exposed in vitro.

The herbicide trifluralin was evaluated for genotoxicity in cultured human peripheral blood lymphocytes. Sister-chromatid exchanges (SCE), chromosome aberrations (CA) and micronuclei (MN) were scored as genetic endpoints. To detect eventual metabolic modification in the genotoxicity of this herbicide, the cultures for SCE and MN demonstration were also treated with S9 fraction. From our results we can conclude that trifluralin was able to exert a weak cytotoxic effect, reducing both the proliferative rate index (PRI) and the cytokinesis block proliferation index (CBPI), and also to induce a slight but statistically significant increase in the frequency of SCE. Under our conditions of testing, no genotoxic effects of trifluralin were observed in the CA and MN assays.

Cells, Cultured↗
PubMed1996

Sister-chromatid exchanges (SCE) induction by inhibitors of DNA topoisomerases in cultured human lymphocytes.

The induction of sister-chromatid exchanges (SCE) in cultured human lymphocytes by four inhibitors of DNA topoisomerases: m-amsacrine, camptothecin, etoposide and nalidixic acid has been evaluated. Although the four compounds apparently increase the frequency of SCE, the effect of nalidixic acid is weak because only a statistically significant positive response was found in one donor at the highest concentration (500 microM). The other compounds tested act as SCE inducers in both donors, camptothecin being the most effective. In addition, the influence of these four topoisomerase inhibitors on the SCE frequency induced by MMC was also analysed. The results reveal that less than additive SCE effect was induced by the combined treatments which could suggest that the process leading to SCE induction by MMC and the four inhibitors of DNA topoisomerases are not totally independent.

Amsacrine↗
PubMed1996

No increase in micronuclei frequency in cultured blood lymphocytes from a group of filling station attendants.

Service station attendants are workers that are definitely exposed to petroleum derivatives. Taking into account that this exposure has been considered to possess genotoxic risk, here we present data on the biomonitoring of a group of 50 service station workers and 43 controls. Micronuclei (MN) from peripheral blood lymphocytes has been considered as the genetic endpoint to be studied and, in addition, data on the concentration of aromatic hydrocarbons at the workplace, urinary metabolites and differential white blood cell count have also been analysed. The results obtained indicate no significant differences between petrol station attendants and controls, when the effects of petrol exposure were investigated by differential white blood cell count and analysis of MN frequencies in phytohaemagglutinin-stimulated lymphocytes. Regarding the urinary metabolites, a significant increase in the phenol level was found in the exposed workers.

Adolescent↗