Environmental Assessment and Evaluation of Oxidative Stress and Genotoxicity Biomarkers Related to Chronic Occupational Exposure to Benzene.

Environmental and occupational exposure to benzene from fuels is a major cause for concern for national and international authorities, as benzene is a known carcinogen in humans and there is no safe limit for exposure to carcinogens. The objective of this study was to evaluate the genotoxic effects of chronic occupational exposure to benzene among two groups of workers: filling station workers (Group I) and security guards working at vehicles entrances (Group II), both on the same busy highway in Rio de Janeiro, Brazil. Sociodemographic data on the workers were evaluated; the concentration of benzene/toluene (B/T) in atmospheric air and individual trans,trans-muconic acid (ttMA) and S-phenylmercapturic acid (S-PMA) were measured; oxidative stress was analyzed by catalase (CAT), glutathione S-transferase (GST), superoxide dismutase (SOD), thiol groups (THIOL) and malondialdehyde (MDA); genotoxicity was measured by metaphases with chromosomal abnormalities (MCA) and nuclear abnormalities, comet assay using the enzyme formamidopyrimidine DNA glycosylase (C-FPG), and methylation of repetitive element LINE-1, CDKN2B and KLF6 genes. Eighty-six workers participated: 51 from Group I and 35 from Group II. The B/T ratio was similar for both groups, but Group I had greater oscillation of benzene concentrations because of their work activities. No differences in ttMA and S-PMA, and no clinical changes were found between both groups, but linearity was observed between leukocyte count and ttMA; and 15% of workers had leukocyte counts less than 4.5 × 109 cells L-1, demanding close worker's attention. No differences were observed between the two groups for THIOL, MDA, MCA, or nuclear abnormalities. A multiple linear relationship was obtained for the biomarkers MCA and C-FPG. A significant correlation was found between length of time in current job and the biomarkers C-FPG, MCA, GST, and MDA. Although both populations had chronic exposure to benzene, the filling station workers were exposed to higher concentrations of benzene during their work activities, indicating an increased risk of DNA damage.

Utilização de biomarcadores de genotoxicidade e expressão gênica na avaliação de trabalhadores de postos de combustíveis expostos a vapores de gasolina / Use of genotoxicity biomarkers and gene expression on the evaluation of gas station attendants exposed to gasoline fumes

Resumo Introdução: a avaliação de uma exposição mensura sua intensidade, frequência e duração, podendo detectar danos precoces que, se ignorados, podem evoluir para um quadro nocivo. Nos campos da saúde ambiental e ocupacional, os biomarcadores de genotoxicidade tem sido largamente utilizados para essa avaliação. Objetivo: identificar, descrever e discutir os principais bioindicadores de genotoxicidade e seu uso conjunto com técnicas de avaliação de expressão gênica em estudos de exposição ocupacional ao benzeno em postos de revenda de combustíveis (PRC). Métodos: revisão bibliográfica de trabalhos publicados entre 1995 e 2015. Resultados: as técnicas identificadas foram: ensaio cometa, estresse oxidativo, micronúcleos, aberrações cromossômicas, polimorfismos, adutos de DNA e proteínas, fatores epigenéticos e expressão gênica. Foi observado que testes de danos genéticos e epigenéticos são utilizados em frentistas de PRC que participam de programas de saúde do trabalhador ou de pesquisas, embora um baixo número de publicações sobre o tema tenha sido identificado. Esse fato talvez possa ser explicado pelos poucos países onde a profissão persiste e pelas limitações para o desenvolvimento de pesquisas nesses países. Conclusão: os bioindicadores de genotoxicidade e as técnicas de expressão gênica são úteis na detecção de dano precoce desta exposição ocupacional e devem ser avaliados em conjunto.

Abstract Introduction: an exposure evaluation measures its intensity, frequencyand duration, detecting premature damage that, if ignored, might develop into a harmful framework. On environmental and occupational health fields, genotoxicity biomarkers have been widely used for this evaluation. Objective: to identify, describe and discuss main genotoxicity biomarkers and their use together with gene expression evaluation techniques in studies concerning occupational exposure to benzene in gas stations (GS). Methods: bibliographical review of studies published between 1995 and 2015. Results: the following techniques were identified: comet assay, oxidative stress, micronuclei, chromosomal aberrations, polymorphisms, DNA and protein adducts, epigenetic factors and gene expression. We observed that genetic and epigenetic damage tests are used in gas station attendants who participate in worker’s health programs or in researches, although a short number of publications on the theme have been identified. This can be explained by the small number of countries where such job still exists and by the limitations for developing research in such countries. Conclusion: genotoxicity biomarkers and gene expression techniques are useful for detecting the premature damage resulting from this occupational exposure and must be jointly evaluated.

Análise epidemiológica dos acidentes ofídicos no município de teresópolis - rj no período de 2007 a 2010 / Epidemiological analysis of snakebites in Teresópolis - RJ from 2007 to 2010

Objetivo: Determinar a epidemiologia e fluxo de informação de notificação dos acidentes ofídicos no município de Teresópolis (RJ). Métodos: Foi realizado um estudo descritivo, quantitativo e retrospectivo dos acidentes e para isso os dados foram coletados por meio de consulta ao banco de dados SINAN NET e das fichas individuais de investigação para notificação de acidentes por animais peçonhentos do SINAN, emitidas pelo Hospital das Clínicas de Teresópolis Costantino Ottaviano no período de 01/2007 ­ 12/2010. Foram avaliadas as seguintes variáveis: tipo, mês e ano do acidente, município de ocorrência, idade, escolaridade, ocupação, sexo, zona de ocorrência, tempo de atendimento, local da picada, manifestações e complicações locais e sistêmicas, tempo de coagulação, classificação, soroterapia, acidente relacionado ao trabalho e evolução do caso. Resultados: Foram analisadas 121 fichas individuais de investigação do município enquanto no SINAN estão registrados 104 casos, indicando uma subnotificação de 14,4%. O gênero Bothrops aparece como responsável pela maioria dos acidentes atendidos com 81,8% dos casos, apresentando sazonalidade maior nos meses entre outubro e março (72,7%), com predominância de acidentados do sexo masculino (78,5%), idade variando entre 20 e 59 anos (80,2%) e maior acometimento dos membros inferiores (68,8%). Conclusões: Concluiu-se que o perfil dos acidentes segue os padrões já descritos na literatura para a região sudeste, mas que existem falhas no preenchimento das fichas individuais de investigação e falta de sintonia entre o processo de coleta de dados e a apresentação dos dados no sistema SINAN NET, apesar deste ser de extrema importância para a informação sobre acidentes com animais peçonhentos (AU).

Objective: To determine the epidemiology and flow notification information of snakebites in the municipality of Teresópolis (RJ). Methods: We conducted a descriptive study, quantitative and retrospective of accidents and that the data were collected by consulting the SINAN NET database and individual research records for venomous animals by accident notification SINAN issued by the Hospital das Teresópolis Costantino Ottaviano clinics in 01/2007 period - 12/2010. The following variables were evaluated: type, month and year of accident occurrence municipality, age, education, occupation, sex, occurrence area, service time, the bite site, demonstrations and local and systemic complications, clotting time, classification, serum therapy, accident related to work and developments. Results: We analyzed 121 individual records municipal research while in SINAN are recorded 104 cases, indicating underreporting of 14.4%. The Bothrops appears as responsible for most injuries treated with 81.8% of the cases, greater seasonality in the months between October and March (72.7%), with predominance of rough male (78.5%), age ranging from 20 to 59 years (80.2%) and increased involvement of the lower limbs (68.8%). Conclusions: We concluded the accident profile follows the standards described in the literature for the southeast region but there are flaws in the filling of individual forms of research and lack of harmony between the data collection process and presentation of data in the SINAN system NET, although this is extremely important for the information on accidents with poisonous animals (AU).

Micronucleus frequency in children exposed to biomass burning in the Brazilian Legal Amazon region: a control case study.

BACKGROUND: The Amazon represents an area of 61% of Brazilian territory and is undergoing major changes resulting from disorderly economic development, especially the advance of agribusiness. Composition of the atmosphere is controlled by several natural and anthropogenic processes, and emission from biomass burning is one with the major impact on human health. The aim of this study was to evaluate genotoxic potential of air pollutants generated by biomass burning through micronucleus assay in exfoliated buccal cells of schoolchildren in the Brazilian Amazon region. METHODS: The study was conducted during the dry seasons in two regions of the Brazilian Amazon. The assay was carried out on buccal epithelial cells of 574 schoolchildren between 6-16 years old. RESULTS: The results show a significant difference between micronucleus frequencies in children exposed to biomass burning compared to those in a control area. CONCLUSIONS: The present study demonstrated that in situ biomonitoring using a sensitive and low cost assay (buccal micronucleus assay) may be an important tool for monitoring air quality in remote regions. It is difficult to attribute the increase in micronuclei frequency observed in our study to any specific toxic element integrated in the particulate matters. However, the contribution of the present study lies in the evidence that increased exposure to fine particulate matter generates an increased micronuclei frequency in oral epithelial cells of schoolchildren.

Avaliação do potencial genotóxico do impacto da queima de biomassa na região sul da Amazônia Legal: estudo de caso na Região de Tangará da Serra, MT / Evaluation of the genotoxic potential impact of biomass burning in southern Amazon: a case study in the Region of Tangara da Serra, MT

A Amazônia Legal brasileira vem sendo impactada com o desenvolvimentoeconômico e o uso do solo não sustentável, em especial, devido à expansão das commodities agrícolas sobre as áreas de florestas e grandes projetos de infraestrutura. A região de Tangará da Serra, Brasil está localizada no Sul da Amazônia Legal, caracterizando-se pela produção de cana-de-açúcar não mecanizada, além de estar na rota de dispersão da pluma dos poluentesgerados pela queima de biomassa no arco do desmatamento. O objetivo deste estudo foi avaliar o potencial de genotoxicidade e mutagenicidade gerado pela exposição aos poluentesatmosféricos liberados a partir da queima de biomassa. Para tanto, foram realizados ensaios cromossômicos por meio da contagem de micronúcleo em sistema vegetal e humano, bem como análise de mutação gênica em sistema bacteriano. Os resultados mostraram uma fortecorrelação entre a exposição à poluição atmosférica da região e o aumento na frequência de micronúcleos em células de mucosa oral e de linhagem pulmonar humana, como também em células vegetais. O estudo também observou uma relação forte e significante entre afrequência de micronúcleos e a morbidade hospitalar por doenças respiratórias em criança. Em conclusão, o presente estudo mostra que o biomonitoramento através de um ensaio sensível e de baixo custo (ensaio de micronúcleo) pode ser uma alternativa para o monitoramento da qualidade do ar nas regiões remotas e pobres.

Genotoxicity evaluation of Moringa oleifera seed extract and lectin.

UNLABELLED: This article reports the genotoxicity assessment of an extract of M. oleifera seed powder and the water-soluble Moringa oleifera lectin (WSMoL) isolated from seeds. The lectin isolated by chitin chromatography showed hemagglutinating activity with different erythrocytes, activity in a broad pH range (4.5 to 9.5), and retention of hemagglutinating activity after being heated to 100 °C. Genotoxicity of the seed extract and WSMoL were assessed using the cell-free plasmid DNA as well as the Salmonella typhimurium (Ames and Kado) assays with TA97, TA98, TA100, and TA102 in the presence or absence of hepatic metabolization. Seed extract at concentration (0.2 µg/µL) recommended to treat water was not genotoxic by Ames, Kado, and cell-free plasmid DNA assays. S. typhimurium strains showed to be sensitive to M. oleifera extract revealing a mutagenic effect at doses higher than 0.6 µg/µL with hepatic metabolization. The extract at doses higher than 0.4 µg/µL, without hepatic metabolization, was mutagenic for TA100 and TA102. WSMoL was nonmutagenic by used assays. The use of high concentrations of the extract may pose a risk to human health and the safe use of M. oleifera seed powder to treat water for human consumption requires more study; however, the purified lectin could be an alternative for water treatment. PRACTICAL APPLICATION: The concentration 0.2 µg/µL of M. oleifera seed extract recommended to treat water for humans did not pose a risk to human health. The mutagenicity detected at concentrations higher than 0.4 µg/µL was not due to WSMoL, lectin isolated from extract.

Avaliação do potencial de mutagenicidade e toxicidade da lectina hipoglicemiante de folha de bauhinia monandra (pata-de-vaca) / Mutagenic and toxic potential assessment of the hypoglycemic lectinfrom bauhinia monandra leave (pata-de-vaca)

As plantas medicinais têm sido usadas desde a antiguidade no tratamento de diversas enfermidades humanas. As folhas de Bauhinia monandra são amplamente utilizadas no Brasil como fitoterápico no tratamento do Diabetes Mellitus. A partir das folhas de B. monandra, foi purificada uma lectina galactose-específica, denominada de BmoLL, que também apresentou uma importante capacidade hipoglicemiante. Seguindo as normas propostas pela Portaria nº 116, de 8 de agosto de 1996 do Ministério da Saúde do Brasil, o trabalho objetivou avaliar o potencial de mutagenicidade e toxicidade da BmoLL, mediante a utilização dos testes com cepas de Escherichia coli da linhagem CC104 (Teste de mutagênesedireta), com cepas de Salmonella typhimurium da linhagem TA (Teste de Kado), com plasmídeo pBCKS (Quebra de DNA plasmidial) e com enzima Exonuclease III (Detecção de sítios abásicos). Os resultados demonstraram que a lectina foi incapaz de aumentar a freqüência de mutação reversa das cepas de S. typhimurium, com e sem ativador metabólico.No entanto, uma diminuição significativa na frequência de mutação espontânea foi observada nas cepas de E. coli, especialmente na deficiente de reparo (CC104mutMmutY), sugerindo um potencial antioxidante da lectina. A BmoLL é incapaz de gerar danos genotóxicos e citotóxicos, com base nas concentrações testadas e nos ensaios realizados.

Medicinal plants have been used since ancient times to treat various human diseases.The leaves of Bauhinia monandra are widely used in Brazil as herbal remedies in the treatmentof Diabetes Mellitus. From the leaves of B. monandra, a galactose-specific lectin was purified,called BmoLL, which also showed a significant hypoglycemic capacity. In accordance with therules proposed by Brazil?s Ministry of Health Decree 116 of 08 August 1996, this study aimedat assessing the potential for toxicity and mutagenicity of BmoLL by means of using tests withEscherichia coli strain CC104 (Forward mutagenesis assay) with Salmonella typhimurium strain TA(Kado test), with plasmid pBCKS (Break occurrences in plasmid DNA) and enzyme exonucleaseIII (Search of abasic sites). Results demonstrated that lectin was unable to increase the frequencyof reverse mutation of strains of S. typhimurium, with and without metabolic activity. However,a significant decrease in the frequency of spontaneous mutation was observed in strains ofE. coli, especially in poor repair (CC104mutMmutY), suggesting an anti-oxidant potential of lectin.BmoLL is unable to generate genotoxic and cytotoxic damage, based on the concentrations andtests performed.

Evaluation of the genotoxic potential of Bauhinia monandra leaf lectin (BmoLL).

Bauhinia monandra, a plant popularly known as "pata de vaca" in Brazil, is widespread in the world and widely used in folk medicine. BmoLL is a galactose-specific lectin obtained and purified from B. monandra leaves, whose hypoglycemiant potential has been recently demonstrated in rats. The present study was performed to investigate the genotoxic potential of BmoLL in a series of cell-free and bacterial assays. We based our test concentrations on those used in popular medicine. The results showed that lectin BmoLL was unable to produce genotoxicity or cytotoxicity in all the assays used. The results also demonstrated that BmoLL did not increase the frequency of reverse mutation in Salmonella typhimurium strains (TA97, TA98, TA100 and TA102), with and without metabolic activation. However, a significant decrease in the spontaneous mutation frequency was observed in Escherichia coli strains (CC104 and CC104mutMmutY), especially in the repair-deficient strain, suggesting an anti-oxidative potential. B. monandra leaf lectin (BmoLL) did not induce cytotoxic or genotoxic effects in the battery of assays used.

[Correlation of hybrid II capture cytologic exam in diagnosis of cervical lesions related to HPV]. / Importância da correlação do exame citológico com a captura híbrida II no diagnóstico de lesões intraepiteliais cervicais relacionadas ao HPV1.

PURPOSE: The aim of the present study was confront the results of the cytological examination with hybrid capture II in the diagnosis of induced cervical intraepithelial lesion-HPV, correlating the cytological findings with biomoleculares. METHODS: The research was carried through in a group of 160 sexually active women who had espontaneamente looked its gynecologists for consultation of routine, having been submitted to the collection of cervicovaginal material for cytology and for examination of hybrid capture II in the Centro de Patologia Clínica and the Hospital e Maternidade Promater, in the city of the Natal-RN. RESULTS: The results had shown to relatively high numbers of positive cases for HPV using hybrid capture II (41.87%) and the cytology (23.75%). The agreement between the two studied methods relatively was raised (59.38%). It was evident also that the viruses with high oncogênico potential had presented found in the compatible cytology with Lesion of low risk (11.88%), followed of Lesion of high risk (NIC II and III); already the viruses with low oncogênico potential were more associates the Lesion of low risk (6.25%), followed of Lesion of high risk. CONCLUSION: The cytology, exactly with its limitations, is an important method in the detention of attributable patologias to the HPV, emphasizing that the molecular method comes to complement it and to consolidate the cytological findings.

[Vulvovaginitis: vaginal pH changes and associated microflora]. / Vulvovaginites: estudo das variações de pH vaginal e microflora associada.

PURPOSE: To establish a correlation between pH vaginal and the microflora associated in carriers of vulvovaginites. METHODS: In the present study, the cytopathological examination and the vaginal flow in a group of 65 sexually active women had been carried through, 20 and 72 years, taken care of in the Laboratório de Citologia Clínica do Departamento de Análises Clínicas e Toxicológicas da Universidade Federal do Rio Grande do Norte, for determination of microorganisms in cervicovaginal sample and of pH in the vaginal flow. RESULTS: Associating pH vaginal with the presence of vulvovaginitis, it was evidenced that the Candida sp. occurred more frequently in pH 4.0, Trichomonas vaginalis in pH 6.0, Gardnerella vaginalis in pH 5.0, coconuts in pH 5.0, bacilli in pH 4.0 and cocos/bacilos in pH 6.0. It was observed that all the patients had presented at least one type of ethiological agent of vulvovaginiti and an associated microflora. CONCLUSION: The joint accomplishment of the cytological examinations and the determination of pH revealed important for directing the microflora associated with the vulvovaginiti, suggesting, of this form, that pH vaginal plays preponderant role how much to the presence of the infectious agents in the vaginal ecosystem.