DNA damage in workers occupationally exposed to pesticide mixtures.

Pesticides are used in agriculture to protect crops but represent at the same time a potential risk to farmers and environment. The aim of this work is the evaluation of 54 subjects occupationally exposed to pesticides and 30 subjects as a control group using the quantification of DNA damage level by means of the alkaline Comet assay and the evaluation of repair processes. Damage index Comet assay (DICA) and damage index repair assay (DIRA) were studied in 27 pesticide applicator workers, 27 non-pesticide applicators and controls. Our results show that both exposed groups revealed significant increase in DICA when compared with controls (P < 0.0001), as well as in DIRA (P < 0.0001). However, the spraying group exhibited a marginally significant difference in DICA (P = 0.05) when years of exposure are considered and a significant difference (P < 0.05) when the personal protective equipment used by individuals was taken as a comparison factor. The influence of confounding factors on the genotoxic effects of occupational exposure to pesticides was investigated and no significant differences were observed considering age, gender, smoking and alcohol consumption in relation to DICA and DIRA. Since DNA damage is an important step in events leading from carcinogen exposure to cancer disease, our study highlights the potential health risk associated with agrochemical exposure in developing countries with vast cultivated areas, such as Argentina.

Genotoxicity and cell death induced by tinidazole (TNZ).

Tinidazole (TNZ), a second-generation 5-nitroimidazole compound chemically related to metronidazole (MTZ), has been widely used throughout Europe and developing countries for the treatment of amoebic and parasitic infections. Despite TNZ's increasing use in therapeutics, scarce experimental reports are available in literature on its potential genotoxicity in human cells. Therefore, the aim of the present study was to achieve a precise characterization of the cytotoxic and genotoxic activities of this nitroimidazole in cultured human lymphocytes at therapeutic concentrations (0.1, 1, 10 and 50 microg/ml of culture) and evaluate the possible cell death mechanism associated with it. The endpoints analyzed included: mitotic index (MI), replication index (RI), sister chromatid exchange (SCE) and chromosomal aberrations (CA). A significant decrease (p<0.0001) in MI as well as an increase in SCE (p<0.0001) and CA (p<0.0001) frequencies were observed. No modifications in RI were found. The results suggest a genotoxic and cytotoxic effect of TNZ related with cell death process. Therefore, we evaluated this mechanism by DNA fragmentation (laddering), fluorescence microscopy using acridine orange/ethidium bromide (AO/EB) staining and flow cytometry propidium iodide (PI). DNA extracts of TNZ-treated cells resulted in nucleosomal DNA ladder pattern after 48 h of cell treatment; meanwhile no differences were detected in untreated cells. This pattern correlated with the observed decrease in cellular viability (p<0.05), morphological evidence of apoptosis and increase in the percentage of nuclei with hypodiploid DNA content of TNZ exposed cultures compared with control (p<0.05). We concluded that TNZ is genotoxic, cytotoxic and is able to modulate cell death through apoptotic mechanisms in the experimental design employed.

Genotoxic and aneugenic properties of an imidazole derivative.

To contribute to a more accurate characterization of the mutagenic and aneugenic effects of thiabendazole (TBZ), a widely used antiparasitic and food preservative drug, the induction of sister chromatid exchanges (SCEs) and mitotic spindle anomalies as cytogenetic end-points were investigated. Studies were carried out in Chinese hamster ovary (CHO) cells and human peripheral blood lymphocytes. A significant dose-dependent increase in SCE frequency was observed in CHO cells with S9-Mix (P < 0.01) in the 50-100 microg ml(-1) dose-range, while in the absence of S9-Mix, an enhancement of the SCE frequency was exhibited at the highest dose (P < 0.01). In CHO-K1 cells a significant increase in mitotic spindle anomalies (P < 0.01) was observed with the highest concentration assayed reflecting the specific effect of TBZ formulation at the microtubule level. Cell proliferation kinetics (CPK) were not modified by the addition of this pharmaceutical product. In human lymphocyte cultures, exposure to 100 microg ml(-1) TBZ formulation resulted in a significant decrease of the mitotic index (MI) (P < 0.003) and changes in the replication index (RI) (P < 0.05).

Anti-genotoxic effect of Aloysia triphylla infusion against acrylamide-induced DNA damage as shown by the comet assay technique.

Aloysia triphylla a perennial, bushy plant originally from South America has long been used in traditional medicine. Its aqueous extract contains considerable amounts of polyphenolic compounds, namely flavonoids and phenolic acids. In view of the interest in natural phenolic compounds as antioxidant in preventive medicine, this study was undertaken to investigate the chemoprotective effects of cedron leaves infusion against the genetic damage induced by acrylamide (AA) by using the alkaline version of the comet assay technique. Mice were separated in nine groups (eight animals each): (I) untreated, (II) negative control, (III) treated with infusion of cedron leaves 5%, 20 days twice a day, (IV) treated with AA (5 mg/kg b.w.), (V) treated with AA (20 mg/kg b.w.), (VI) treated with AA (30 mg/kg b.w.), (VII) treated with AA (50 mg/kg b.w.), (VIII) pretreated with infusion and treated with AA (50 mg/kg b.w.) and (IX) positive control (cyclophosphamide, 20 mg/kg b.w.). Three hundred blast cells were digitally evaluated per animal from three different slides (100 each). Media of tail moment (TM) values were analyzed by ANOVA test. No statistical differences (p>0.05) were found between untreated animals, negative control and infusion-treated mice. A single dose of AA-induced genetic damage as revealed by a statistically significant increase in TM values (p<0.01). Pretreatment with infusion prior to AA injection significantly reduces the capacity of AA to induce genetic damage. In these conditions, tail moments values did not differ from data obtained in negative control (p>0.05) and exhibit statistical differences from animals treated only with AA (p<0.01). Cell viability was at least 90% in all cases as measured by the trypan blue exclusion method. The ferric reducing ability of plasma (FRAP) method reveals that the plasma of infusion-treated mice has a significantly higher antioxidant capacity than plasma from controls (p<0.01). The results suggest that the infusion could exerts an in vivo chemo protective action, probably due to its scavenging potency towards free radicals.

Argentine folk medicine: genotoxic effects of Chenopodiaceae family.

Chenopodium ambrosioides L. and Chenopodium multifidum L. (Chenopodiaceae), common name: Paico, are medicinal plants. They are aromatic shrubs growing in South America. For centuries, they have been used due to its medicinal properties. However, there are few reports in literature about the genotoxic effects of these plants. There for, the aim of these work is the evaluation of genetic damage induced by decoction and infusion of this plants which were assayed in different concentrations (1, 10, 100, 1,000 microL extract/mL culture), by addition of the extract to human lymphocyte cell cultures, negative controls were included. The endpoints evaluated were chromosomal aberrations (CA), sister chromatid exchanges (SCE), cell proliferation kinetics (CPK) and mitotic index (MI). The repeated measure analysis of variance was used for statistic evaluation of the results. The results showed: (a) statistical increase in the percentage of cells with CA and in the frequency of SCE when cultures were exposed to both aromatic plants, (b) a decrease in MI of both Paicos assayed, although no modification in the CPK values was observed, (c) no effect was noticed in the analysis of Chenopodium album L., which was used as negative control of the essential oil. These results suggest a cyto and genotoxic effect of Chenopodium ambrosioides and Chenopodium multifidum aqueous extracts related to the essential oil of the plant (as Chenopodium album did not perform).

Teratogenic evaluation of metronidazole and ornidazole using Drosophila melanogaster as an experimental model.

BACKGROUND: Drosophila and vertebrates show similarities that suggest that the mechanisms involved in the induction of developmental defects may be similar in both. Therefore, Drosophila has been proposed as a useful, rapid, and economical model in the preliminary screening for teratology studies. The objective of the present study was to investigate the effect of metronidazole (MTZ) and ornidazole (ONZ) on the developmental stages of Drosophila melanogaster. METHODS: Samarkand wild-type females were allowed to lay eggs for 24 hr in media containing MTZ or ONZ at concentrations of 0, 500, 1000, and 2000 microg/ml. When larvae completed their development, the emerging flies were counted and examined for morphological abnormalities. RESULTS: After the analysis of 400-1000 flies for each concentration, ONZ-treated flies did not show an incidence of malformations above control values, although a significant high number of individuals with reduced body size was observed (p < 0.005, chi2 test). On the other hand, the 1000- and 2000-microg/ml MTZ-treated series presented higher frequencies of total abnormalities than did concurrent and historic controls (p < 0.05, chi2 test), indicating an MTZ effect during developmental morphogenesis. CONCLUSIONS: These findings contribute to the characterization of both nitroimidazoles, which are widely used, especially in underdeveloped countries. At the same time, this Drosophila bioassay is sensitive enough to detect differential effects of MTZ and ONZ (abnormalities vs. growth effects), showing specificity and selectivity.

Toxicogenetic evaluation of metronidazole in the treatment of women infected with Trichomonas vaginalis.

Samples of peripheral blood were collected once from non-smoking women who were healthy (controls) and twice (immediately before and immediately after 7 days of treatment with metronidazole at 500 mg/day) from non-smoking women infected with Trichomonas vaginalis. Lymphocyte cultures were prepared and used, in toxicogenetic studies, to determine the frequency of sister-chromatid exchange (SCE), the mitotic index (MI), and the replication index (RI) for each sample. MTZ treatment of the infected women led to an increase in the frequency of SCE (P <0.001), a decrease in the MI (P <0.003), and a modification in the kinetics of cell proliferation, with a decrease in the RI (P <0.0006). The differences seen between the results for the controls and those for the infected women, before and after MTZ treatment, may be attributed to the presence of the parasite, to the treatment itself, and/or to variation in the host's response to infection with T. vaginalis.

Cytogenetic evaluation of two nitroimidazole derivatives.

5-Nitroimidazoles are a well-established group of antiprotozoal and antibacterial agents. Thanks to their antimicrobial activity these chemotherapeutic agents inhibit the growth of both anaerobic bacteria and certain anaerobic protozoa such as Trichomonas vaginalis, Entamoeba histolytica and Giardia lamblia. The aim of the present study is to achieve a precise characterization of the genotoxic activity of these compounds and to establish the value of cytogenetic assays in order to determine the effect of these drugs, at therapeutic doses, to settle an improved risk assessment. Two nitroimidazole were studied, metronidazole and ornidazole, at four different concentrations (0.1, 1, 10 and 50 microg/ml of peripheral blood lymphocyte culture). Endpoints analyzed included: mitotic index (MI), replication index (RI), sister chromatid exchange (SCE) and chromosomal aberrations (CA). An analysis of variance test (ANOVA) was performed to evaluate the results. A significant decrease (P<0.0001) in MI as well as an increase in SCE (P<0.0001) and CA (0.0001) frequencies for both drugs was observed. No modifications in RI were found. The results suggest a genotoxic and cytotoxic effect of MTZ and ONZ in human peripheral blood cultures in vitro.

The behavior of sex chromosomes in two human X-autosome translocations: failure of extensive X-inactivation spreading.

Two patients, one adult male and one infant girl, bearing different X-autosome translocations, were studied with cytogenetical, ultrastructural and chromosome-painting techniques. The adult male, is a carrier of a reciprocal, balanced translocation involving the X and #2 chromosomes: 46,Y,t(X;2) (q13;p21). This man showed infertility with spermatogenesis arrest at the spermatocyte stage. Synaptonemal complex analysis at pachytene showed the quadrivalent structure and the putative breakage points. Sex-chromatin condensation did not spread towards the autosomal regions of the quadrivalent. The female infant showed diminished body growth and multiple somatic anomalies. She is a 45,Xp-,t(X;21)(p11;p13) carrier, an unbalanced translocation involving chromosomes X and #21, which leads to a monosomy of almost all Xp. The translocated #21 is practically complete, and its centromere is the active one in the rearranged product. The analysis of interphase nuclei with the X-centromere probe shows that the Xq region of the rearranged chromosome is the late -replicating and inactive element. However, X-inactivation does not spread to the attached #21, as shown by the R-banding pattern. Thus, both in the male adult and in the female infant there is a barrier to the spreading effect of X-chromosome inactivation, which is probably due to different mechanisms.

The behavior of sex chromosomes in two human X-autosome translocations: failure of extensive X-inactivation spreading

Two patients, one adult male and one infant girl, bearing different X-autosome translocations, were studied with cytogenetical, ultrastructural and chromosome-painting techniques. The adult male, is a carrier of a reciprocal, balanced translocation involving the X and #2 chromosomes: 46,Y,t(X;2) (q13;p21). This man showed infertility with spermatogenesis arrest at the spermatocyte stage. Synaptonemal complex analysis at pachytene showed the quadrivalent structure and the putative breakage points. Sex-chromatin condensation did not spread towards the autosomal regions of the quadrivalent. The female infant showed diminished body growth and multiple somatic anomalies. She is a 45,Xp-,t(X;21)(p11;p13) carrier, an unbalanced translocation involving chromosomes X and #21, which leads to a monosomy of almost all Xp. The translocated #21 is practically complete, and its centromere is the active one in the rearranged product. The analysis of interphase nuclei with the X-centromere probe shows that the Xq region of the rearranged chromosome is the late -replicating and inactive element. However, X-inactivation does not spread to the attached #21, as shown by the R-banding pattern. Thus, both in the male adult and in the female infant there is a barrier to the spreading effect of X-chromosome inactivation, which is probably due to different mechanisms.

The behavior of sex chromosomes in two human X-autosome translocations: failure of extensive X-inactivation spreading

Two patients, one adult male and one infant girl, bearing different X-autosome translocations, were studied with cytogenetical, ultrastructural and chromosome-painting techniques. The adult male, is a carrier of a reciprocal, balanced translocation involving the X and #2 chromosomes: 46,Y,t(X;2) (q13;p21). This man showed infertility with spermatogenesis arrest at the spermatocyte stage. Synaptonemal complex analysis at pachytene showed the quadrivalent structure and the putative breakage points. Sex-chromatin condensation did not spread towards the autosomal regions of the quadrivalent. The female infant showed diminished body growth and multiple somatic anomalies. She is a 45,Xp-,t(X;21)(p11;p13) carrier, an unbalanced translocation involving chromosomes X and #21, which leads to a monosomy of almost all Xp. The translocated #21 is practically complete, and its centromere is the active one in the rearranged product. The analysis of interphase nuclei with the X-centromere probe shows that the Xq region of the rearranged chromosome is the late -replicating and inactive element. However, X-inactivation does not spread to the attached #21, as shown by the R-banding pattern. Thus, both in the male adult and in the female infant there is a barrier to the spreading effect of X-chromosome inactivation, which is probably due to different mechanisms.(AU)

[Pneumonia due to mycoplasma pneumoniae in a primary health care district]. / Neumonía por Mycoplasma pneumoniae en una zona básica de salud.

OBJECTIVE: Most studies of atypical pneumonia due to Mycoplasma pneumoniae have been performed in hospitalized patients. The aim of this study was to determine the epidemiological, clinical and radiological characteristics as well as the evolution of patients with atypical pneumonia due to Mycoplasma pneumoniae in an out-of-hospital setting. METHODS: A prospective observational study was conducted in 31 patients with atypical pneumonia due to Mycoplasma pneumoniae in the pediatric population of a primary health care district from May to July 1996. In all patients serological confirmation of infection by Mycoplasma pneumoniae was obtained using a complement-fixation test. RESULTS: The age of the children in the study ranged from 4-13 years. The most common clinical manifestations were cough (93 %), high fever (84 %) and pharyngitis (48 %). The most frequent auscultatory finding was crackles (93 %), which were bilateral in 14 patients. Clinical-radiological dissociation was found in 16 % of the children; 27 (87 %) showed radiological alterations. No characteristic radiological pattern was detected because alveolar and interstitial alterations were equally frequent, with a predominance of lung base involvement (67.7 %). In all patients response to macrolides was excellent, fever abated within 48-72 h and the remaining symptoms progressively improved. CONCLUSIONS: Atypical pneumonia due to Mycoplasma pneumoniae normally presents in children aged more than 5 years old or in adolescents. No signs, symptoms, or radiological patterns clearly indicate the etiology. However, because of its frequency, this disease should be suspected when school-aged children or adolescents present a pneumonia syndrome. Macrolides administration

Evaluation of genetic damage induced by a nitroimidazole derivative in human lymphocytes: Tinidazole (TNZ).

One of the useful drugs in the treatment against infestations caused by Trichomonas vaginalis, Entamoeba histolytica and Giardia lamblia is Tinidazole (TNZ) 1-[2-(ethylsulfonyl) ethyl]-2-methyl-5-nitroimidazole) (Gilman R.H., Marquis G.S., Miranda E., Vestegui M., Martinez H., 1988. Rapid reinfection by Giardia lamblia after treatment in a hyperendemic third world community. Lancet i, 343-345). We decided to evaluate the potential genetic damage induced by TNZ using different biological biomarkers such as the mitotic index (MI), sister chromatid exchange (SCE) and cell proliferation kinetics (CPK). We observed a significant decrease (P<0.0005) in the MI as well as an increase (P<0.0005) in SCE frequency and no modifications in the replication index (RI). The results obtained suggest a potential genotoxic and cytotoxic effect of TNZ in human peripheral blood cultures in vitro.

Embryolethality induced by metronidazole (MTZ) in Rattus norvegicus.

Parasitic illnesses is increasing all over the world, especially in developing countries, and metronidazole (MTZ) is the therapeutic agent usually administered to children as well as adults at the reproductive age. In this work, we propose an evaluation of MTZ in order to analyze the potential reproductive damage in females by using Rattus norvegicus (Sprague-Dawley) as an animal model. Adult female rats were mated after MTZ treatments, and they were sacrificed at 21 days of gestation. Different types of damage were evaluated by using mortality, phenotypic abnormalities and reproductive capacity as parameters, and were studied and scored in 70 adult specimens (450 g/bw). They were divided into five groups: a) untreated females as a control group; females treated with b) DMSO as a solvent control group or c) 500 mg/kg/bw of MTZ per day for 7 days as therapeutic dose (TD); d) a half therapeutic dose (HD); and e) a double therapeutic dose (DD). Pre-implantation death in MTZ-treated groups was not significantly different from controls. However, drug treatments significantly increased the frequency of post-implantation deaths and the dominant lethals were ranged between 12.0 % and 17.8 %.

[Vertical transmission of hepatitis C virus]. / Transmisión vertical del virus de la hepatitis C.

OBJECTIVES: Few studies have been published on vertical transmission of hepatitis C virus (HCV), although it is the most common cause of hepatitis C in children. We aimed to determine the rate of vertical transmission of HCV in at risk neonates and to assess the effect of possible risk factors. METHODS: A prospective follow-up study was conducted in 35 children of seropositive mothers during an 18-month period (July 1997-January 1999). Testing for anti-HCV antibodies was performed with third generation enzyme linked immunoadsorbent assay. HCV-RNA was qualitatively analyzed with reverse transcriptase polymerase chain reaction (RT-PCR) and hepatic enzyme studies. RESULTS: All the 35 children studied were positive for HCV antibodies at birth. The children became HCV negative at a mean age of 6 months. HCV infection was detected in two children (5.7%). The mother of one of these children had both HCV and human immunodeficiency virus (HIV) infection. Among the 35 seropositive mothers, a risk factor for percutaneous transmission of HCV (parenteral injection, drug addiction, or previous transfusions) was detected in 19(54%) and HIV coinfection was found in 9(26%). CONCLUSIONS: The present study is consistent with other studies that found a vertical HCV transmission rate of approximately 5%, with a greater risk if the mothers had HCV/HIV coinfection or parenteral risk factors. Studies with greater numbers of subjects are required to determine the prevalence of HCV in expectant mothers and the precise rate of vertical transmission. Infected children should be followed up to evaluate the repercussions of HCV infection.

The behavior of sex chromosomes in two human X-autosome translocations: failure of extensive X-inactivation spreading.

Two patients, one adult male and one infant girl, bearing different X-autosome translocations, were studied with cytogenetical, ultrastructural and chromosome-painting techniques. The adult male, is a carrier of a reciprocal, balanced translocation involving the X and #2 chromosomes: 46,Y,t(X;2) (q13;p21). This man showed infertility with spermatogenesis arrest at the spermatocyte stage. Synaptonemal complex analysis at pachytene showed the quadrivalent structure and the putative breakage points. Sex-chromatin condensation did not spread towards the autosomal regions of the quadrivalent. The female infant showed diminished body growth and multiple somatic anomalies. She is a 45,Xp-,t(X;21)(p11;p13) carrier, an unbalanced translocation involving chromosomes X and #21, which leads to a monosomy of almost all Xp. The translocated #21 is practically complete, and its centromere is the active one in the rearranged product. The analysis of interphase nuclei with the X-centromere probe shows that the Xq region of the rearranged chromosome is the late -replicating and inactive element. However, X-inactivation does not spread to the attached #21, as shown by the R-banding pattern. Thus, both in the male adult and in the female infant there is a barrier to the spreading effect of X-chromosome inactivation, which is probably due to different mechanisms.

A nursing approach: towards the integral health care of renal patients.

In February 1993, after several multicentre meetings held together with nephrologists, the decision was taken to create the Peritoneal Dialysis Nursing Group for the Central Area; having as its primary purpose the provision of integrated medical care to renal patients.