Algumas áreas do canal de Piaçaguera, localizado na Baixada Santista, SP-Brasil, apresenta alto nível contaminação por hidrocarbonetos policíclicos aromáticos (HPA) entre outros contaminantes nos sedimentos. O objetivo do presente estudo foi verificar diferentes biomarcadores de exposição em bile e de efeitos genotóxicos em sangue de peixes, coletados in situ, que poderiam ser utilizados como ferramenta de monitoramento. O canal de Bertioga foi selecionado como região de referência. Embora não seja uma região sem nenhuma interferência de contaminantes, os sedimentos apresentam baixos valores para HPA, metais e atividade mutagênica. A espécie Mugil curema foi selecionada por ser freqüentemente encontrada em ambas as regiões de estudo. Observamos que os peixes coletados no canal de Piaçaguera apresentaram maior porcentagem de micronúcleos e danos ao DNA (ensaio cometa) no sangue quando comparados com a região de referência. Para o teste de micronúcleo, realizamos leituras de 1000 e 4000 células/indivíduo e não foram observadas diferenças estatísticas, sugerindo que o número de 1000 células poderia ser suficiente para gerar dados confiáveis para a espécie Mugil curema. A avaliação de mutagenicidade da bile foi realizada pelo teste Salmonella/microssoma combinado à extração de bile utilizando Blue rayon (BR) com as linhagens TA98, TA100 e YG1041 com e sem S9 (ativação metabólica). As maiores respostas mutagênicas foram observadas para os peixes coletados no canal de Piaçaguera. A mutagenicidade com a linhagem YG1041 na presença de S9 foi mais elevada quando comparado à mutagenicidade observada na ausência de S9 e também em comparação com a sua linhagem parental TA98, indicando a provável presença de compostos da classe das aminas aromáticas na bile como responsáveis pelos efeitos observados. A quantificação de metabólitos equivalentes de HPA foi realizada por HPLC/fluorescência na bile bruta e embora os resultados não se correlacionem com a mutagenicidade, altos níveis também foram encontrados para os peixes coletados no canal de Piaçaguera em comparação com Bertioga. O protocolo de amplificação do gene ras para a espécie Mugil curema foi estabelecido e os genes foram seqüenciados para verificação da presença de mutações. Não foram observadas mutações nos poucos peixes analisados e mais estudos são necessários para verificar a utilização deste biomarcador. O teste de micronúcleo, ensaio cometa e análise de mutagencidade em bile extraída por Blue rayon parecem ser ferramentas biológicas adequadas para monitoramento da qualidade ambiental de estuários contaminados. / Some areas of the Piaçaguera channel at Baixada Santista, SP, Brazil present high levels of polycyclic aromatic hydrocarbons (PAHs) among other contaminants in the sediment. The aim of the present study was to verify if biomarkers of exposure in bile and effect in blood of fishes collected in the field could be used as a monitoring tool. We selected Bertioga channel as a reference area. Although it is not a pristine area, the sediment presents low values of PAHs, metals and mutagenic activity. We selected the fish species Mugil curema, because they are frequently found in both areas during the entire year. We observed that the fish collected at the Piaçaguera channel showed a higher percentage of micronuclei and DNA damage (comet assay) in blood when compared to the Bertioga channel. The micronucleus readings were done in 1000 and 4000 cells/fish and no statistical difference was observed, suggesting that the number of 1000 cells would be sufficient to generate reliable data for the species Mugil curema , in the studied area. The mutagenicity of the bile was performed using the Salmonella/microsome assay combined with Blue rayon (BR) extraction with TA98, TA100 and YG1041 strains with and without rat liver S9. We observed that the mutagenic responses were higher in fish collected in Piaçaguera. The mutagenicity in YG1041 with S9 in fishes collected in the Piaçaguera channel was higher when compared to the mutagenicity observed in the absence of S9 and also higher than the response with the parental strain TA98. The results suggest that mutagenic polycyclic compounds, probably from the class of the aromatic amines are causing the observed effect. PAH metabolites-equivalent were also determined using HPLC/fluorescence in crude bile, and although the results did not correlate with the mutagenicity, higher levels were observed in fish collected in the Piaçaguera channel when compared to Bertioga. A protocol for amplification of the ras gene for the Mugil curema species was established and gene sequenced. No mutations were observed in the liver of the few fishes analyzed, and more studies are required to verify the utility of this biomarker in this fish. The micronuclei, comet assay and mutagenicity in bile extracted with blue rayon seem to be suitable biological tools to monitor the environmental quality of contaminated estuaries.

The occupational exposures of sewage workers are complex and variable, and include a great variety of biological and chemical agents. Previous research has focused mostly on infections and various symptoms among sewage workers, e.g. abdominal and respiratory symptoms. At several sewage plants in Sweden, concern arose about occupational cancer, specifically cancer of the stomach, the kidney, and the lung. The aim of this study was to study the cancer incidence among municipal sewage workers, some exposures that might be connected with cancer risk, and self reported abdominal and respiratory symptoms. In a cohort of municipal sewage workers there was no increase in the overall incidence of cancer when compared with the general population. However, there was a slight increase in the incidence of prostate cancer, but not in the sites of original concern among the workers. Infection by the gastric carcinogen Helicobacter pylori (determined from the presence of IgG antibodies in serum against H pylori) was no more prevalent in sewage workers than in comparable referents. Neither were sewage workers more exposed to genotoxic agents than comparable referents, as measured by the alkaline single cell gel electrophoresis (SCG) assay performed on peripheral lymphocytes. There was no increase in the three-month prevalence of abdominal symptoms when compared with other municipal workers. Specifically, there was no difference in prevalence of the common disorders dyspepsia and irritable bowel syndrome. Sewage workers reported adult bronchial asthma significantly more than the referents.

Pesticides are among the most pervasive environmental contaminants and they are an important potential risk for human health. Agricultural workers are constantly exposed to pesticide spray, drift and residues in the soil and foliage. Many agricultural pesticides are readily absorbed by the body, through contact with the skin, the respiratory track, the eyes, and the gastrointestinal system. Multiple studies have reported a strong association between pesticide exposure and various health outcomes including cancer. Oxidative stress and DNA damage have been proposed as mechanisms linking pesticide exposure to health effects and neurological diseases. The focus of the present translational study is to examine the relationship between human exposure to the organophosphate pesticide azinphos methyl (AZM) and oxidative stress by measuring biomarkers of oxidative stress in biological fluids (i.e., urine, serum) and peripheral blood lymphocytes (PBLs) of agricultural workers. The findings from these field studies will be validated in vitro by examining cultures of human lymphocytes treated with AZM for similar biomarkers of oxidative stress. Since the collection of PBLs from study participants is highly invasive and not suitable for studies involving younger subjects, we also examined buccal cells for biomarkers of oxidative stress (i.e., DNA damage) as a more universal source of human tissue to assess oxidative stress in pesticide exposed individuals. We demonstrated in this study that AZM induces oxidative stress and causes DNA damage in human tissues. Agricultural workers who had been exposed to AZM showed elevated serum levels of lipid peroxides, increased urinary levels of 8-OH-dG, and lymphocytes from these individuals showed increased DNA damage and associated changes in oxidative DNA repair enzymes. Biomarkers of oxidative stress were also elevated in human lymphocytes treated with physiologically relevant concentrations of AZM. In cultures of human lymphocytes, AZM caused a concentration-dependent loss of viability and associated increases in ROS and a reduction in intracellular GSH. We also demonstrated that viable leukocytes from the oral cavity can be readily obtained from humans and these buccal cells can be used to assess DNA damage following exposure to occupational and environmental genotoxicants. We also noted that oral leukocytes are especially sensitive to cryopreservation with DMSO and thus, these cells must be cryoprotected with 5% DMSO to preserve the viability of these cells for subsequent biochemical studies. In summary, these in vivo and in vitro studies demonstrated that AZM induces oxidative stress in a dose-dependent matter and that oral lymphocytes are a good source of human tissue for assessing DNA damage and possibly other biochemical changes. The possible health implications of the variations in these biomarkers of oxidative stress and DNA damage are undetermined. Yet the findings from these studies have provided a strong foundation for determining the mechanism by which pesticide induce oxidative stress, to explore the putative relationship between pesticide-induced oxidative stress and disease (e.g. cancer, neurodegenerative disorders) and determine whether tissue damage in humans is brought about by direct or by indirect action of organophosphate pesticides. / Graduation date: 2010

Ionisierende Strahlung verursacht in Abhängigkeit von den strahlenphysikalischen Eigenschaften der Radionuklide Zellschäden unterschiedlicher Komplexität. An humanen Lymphozyten wurde untersucht, ob die biologische Wirksamkeit von Alpha- und Betastrahlung sowie der Einfluss von Doxorubicin der Qualität des Strahlenschadens zugewiesen werden kann. Die DNA-Schäden und deren Reparatur wurden mit zellbiologischen Methoden quantifiziert.

Biologische Systeme sind in der Lage sich an eine Niedrig-Dosis-Bestrahlung anzupassen und eine geringere Sensitivität gegenüber einer nachfolgenden Hoch-Dosis-Bestrahlung zu entwickeln. Dieses Phänomen wird als adaptive Antwort bezeichnet und wurde nach der Bestrahlung mit externen Strahlungsquellen wiederholt in vivo und in vitro untersucht. Im Gegensatz dazu gibt es für die Bestrahlung mit offenen Radionukliden keine systematischen und vergleichenden Untersuchungen. Im Mittelpunkt dieser Arbeit standen der Nachweis sowie die Analyse der adaptiven Antwort an PC Cl3-Zellen nach Bestrahlung mit den offenen Radionukliden Re-188 und Tc-99m. Die Zellschädigung wurde mit dem alkalischen Komet-Assay, zur Bestimmung des initialen DNA-Schadens und dem Koloniebildungstest, zur Ermittlung des klonogenen Überlebens, untersucht. Zur Aufklärung von möglichen Regulationsmechanismen der adaptiven Antwort wurde die Induktion und Reparatur von DSB mit dem gamma-H2AX-Immunfluoreszenz-Assay und die intrazelluläre Radionuklidaufnahme betrachtet. In dieser Arbeit erfolgte erstmals eine systematische Untersuchung der adaptiven Antwort nach Bestrahlung mit offenen Radionukliden in vitro. Insgesamt zeigen diese Ergebnisse, dass nach Bestrahlung mit offenen Radionukliden eine adaptive Antwort ausgebildet wird. Diese ist von der Strahlenqualität während Vor- und Folgebestrahlung sowie der Art der DNA-Schädigung und den initiierten Reparaturprozessen abhängig. Weiter Einflussfaktoren sind die Erholungszeit, die Vorbestrahlung (Dosis, Strahlenqualität) und die Art des Schadensnachweises. Neben den bekannten Regulationsmechanismen wurde erstmals die Reduktion der intrazellulären Radionuklidaufnahme als weitere mögliche adaptive Antwort beschrieben.

The coastal areas are subject to multiple anthropogenic pressures including chemical pollution that can pose a real risk to the sustainability of aquatic species. The Arcachon Bay, macrotidal lagoon located on the French Atlantic coast, is the important ecosystem for oyster farming. But for several years, the oyster farms face lower recruitment and high mortality of oyster spat. Chemical contamination of the environment as a factor that may contribute to the observed effects on oysters has so far not been investigated.The present thesis aimed at evaluating through different approaches, of the potential toxicity of heavy metals and pesticides representative of the Arcachon Bay contamination on the early life stages of the Pacific oyster, Crassostrea gigas. Embryotoxicity, genotoxicity and expression levels of eleven targeted genes were studied. Firstly, different pesticides (S-metolachlor, irgarol, and diuron) and metals (copper and cadmium) were separately tested to determine their spectrum of effects. It were shown that exposure of gametes and embryos of oyster to environmental concentrations of pesticides and copper increased developmental abnormalities and DNA damage, and reduced fertilization success and affected offpring quality. Cadmium, meanwhile, showed no embryotoxic and genotoxic effects at the concentrations found in the Arcachon Bay. Metabolites of metolachlor, metolachlor ESA and metolachlor OA, are found in the Arcachon Bay at higher concentrations than their parent compound. The results showed that these metabolites were less embryotoxic and genotoxic on oyster embryos and spermatozoa than metolachor. Significant changes in expression of genes involved in antioxidant defense were observed for oyster larvae exposed to metolachlor and metolachlor ESA. Toxicity of mixtures of pesticides representative of the Arcachon Bay contamination with and without copper was then evaluated. Exposures of oyster embryos to these mixtures lead to development defects, DNA damage and changes in the expression of genes involved mainly in oxidative stress responses. Finally, mapping of toxicity of sediments from the Arcachon Bay was conducted for four seasons of 2011 with the oyster embryo-larvae assay. Sediments collected from Arguin exhibited low toxicity, regardless any season. In contrast, sediments from Le Tès showed higher toxicity in spring and summer seasons compared to winter season.From this work, it can be hypothesized that chemical contamination of the Arcachon Bay represents a threat for oyster reproduction and development.

Modeliais grįsta sistemų architektūra (MDA) yra „Object Management Group“ (OMG) konsorciumo iniciatyva apibrėžti naują požiūrį į programų sistemų kūrimą remiantis modeliais ir automatizuota jų transformacija į programinį kodą. Siekdama standartizuoti šį požiūrį, OMG patvirtino visą eilę standartų, bet esminiai MDA principai ir praktikos glūdi modeliais grįstame sistemų kūrimo stiliuje, kuris yra fundamentalus programų sistemų inžinerijoje. MDA idėjos, pradžioje sukėlusios didelį entuziazmą IT bendruomenėje, ilgainiui peraugo į skepticizmą ir kai kurie autoriai atvirai pradėjo abejoti modeliais grįsto sistemų kūrimo perspektyva. Šiame darbe analizuojamos praktinio MDA taikymo programų sistemų kūrimo procese aspektai ir galimybės, analizuojami galimi sprendimai ir kliūtys, dėl kurių MDA požiūris gali būti sunkiai pritaikomas. Įvairių autorių įvardijami praktiniai MDA taikymo programų sistemų kūrimo procese sunkumai, didelės tam reikalingos investicijos, pastangos ir resursai, tinkamų instrumentų trūkumas, didelė standartų įvairovė ir sudėtingos technologijos iš dalies lėmė išaugusį skepticizmą MDA paradigmai. MDA apibrėžia naujus sistemų kūrimo principus ir standartais apibrėžia technologijas, kurios pagrindžia šiuos principus. Tai neišvengiamai įtakoja patį programų sistemų kūrimo procesą, kuris turi pasipildyti naujomis veiklomis, praktikomis ir technologijomis, kaip kurti sistemas taikant MDA požiūrį. Šiame kontekste būtinas tam tikras pragmatinis požiūris į MDA ir su juo... [toliau žr. visą tekstą] / Model Driven Architecture (MDA) is an approach to using models in software development, which states that models and model-based transformations are a key part of effective automated software development. The Object Management Group (OMG) has defined standards for representing MDA models, but the principles and practice of MDA are rooted in model-based styles of development that have been fundamental to software engineering from its earliest days. Unfortunately, early enthusiasm for Model Driven Architecture (MDA) has dissipated to the point that many people are openly skeptical of the value of any model-driven approach. This paper examines the practical realities of MDA, difficulties and challenges in adopting an MDA approach to software engineering process. While MDA requires additional efforts and high investment to be adopted in software engineering process, it doesn’t provide any means or guidelines for this. This paper argues that to be successful, a pragmatic MDA approach must be executed in context of a sound Enterprise Architecture providing an integrated business architecture and governance structure that enables an organization to respond to business requirements quickly and appropriately.

Objectives: The aim of this study was to determine if exposure to volatile organic compounds can lead to DNA damage and impaired DNA repair capacity. Nail cosmetics is a fast growing industry around the world where employees and clients are subjected to various chemical substances which may be harmful to their health: such as formaldehyde, toluene, acetone, xylene, ethylmethacrylate, methylmethacrylate and n–buthyl acetate. These chemicals have the potential to be harmful to their health and exposure to these chemicals should be actively controlled. Formaldehyde is classified as a human carcinogen by the IARC, whereas, toluene and xylene are group three carcinogens, classified in 1999 (not classified as carcinogenic to humans), and various studies have linked DNA damage and impaired DNA repair to the above mentioned substances. Methods: Fifteen nail technicians were monitored by means of personal air sampling, measuring formaldehyde, toluene, xylene, acetone and ethylmethacrylate exposure. Fifteen unexposed subjects were chosen and matched for age and smoking habits with the exposed group. Heparinised blood samples were obtained from each test subject with which the Comet Assay was performed on lymphocytes to determine DNA damage and repair ability. Results: Exposure to ethylmethacrylates and methylmethacrylates leads to DNA damage. Methylmethacrylate causes DNA damage by specifically targeting pyrimidine (fpg) bases. N–buthyl acetate, xylene and acetone exposure impaired DNA repair capacity. The exposed group showed signs of Class III and Class IV DNA damage, whereas the control group had little Class III damage and no indication of Class IV damage. The overall DNA repair ability of the nail technicians was slightly impaired when compared to that of the control group, which is in concurrence with previous studies. Smoking habits and age did not show significant influences on the level of DNA damage and repair when compared with the control group. Conclusion: Exposure to volatile organic compounds such as ethylmethacryale and methylmethacrylate may lead to DNA damage and altered DNA repair in some individuals, although further studies are recommended. / Thesis (M.Sc. (Occupational Hygiene))--North-West University, Potchefstroom Campus, 2011.

Objectives: The aim of this study was to determine if exposure to volatile organic compounds can lead to DNA damage and impaired DNA repair capacity. Nail cosmetics is a fast growing industry around the world where employees and clients are subjected to various chemical substances which may be harmful to their health: such as formaldehyde, toluene, acetone, xylene, ethylmethacrylate, methylmethacrylate and n–buthyl acetate. These chemicals have the potential to be harmful to their health and exposure to these chemicals should be actively controlled. Formaldehyde is classified as a human carcinogen by the IARC, whereas, toluene and xylene are group three carcinogens, classified in 1999 (not classified as carcinogenic to humans), and various studies have linked DNA damage and impaired DNA repair to the above mentioned substances. Methods: Fifteen nail technicians were monitored by means of personal air sampling, measuring formaldehyde, toluene, xylene, acetone and ethylmethacrylate exposure. Fifteen unexposed subjects were chosen and matched for age and smoking habits with the exposed group. Heparinised blood samples were obtained from each test subject with which the Comet Assay was performed on lymphocytes to determine DNA damage and repair ability. Results: Exposure to ethylmethacrylates and methylmethacrylates leads to DNA damage. Methylmethacrylate causes DNA damage by specifically targeting pyrimidine (fpg) bases. N–buthyl acetate, xylene and acetone exposure impaired DNA repair capacity. The exposed group showed signs of Class III and Class IV DNA damage, whereas the control group had little Class III damage and no indication of Class IV damage. The overall DNA repair ability of the nail technicians was slightly impaired when compared to that of the control group, which is in concurrence with previous studies. Smoking habits and age did not show significant influences on the level of DNA damage and repair when compared with the control group. Conclusion: Exposure to volatile organic compounds such as ethylmethacryale and methylmethacrylate may lead to DNA damage and altered DNA repair in some individuals, although further studies are recommended. / Thesis (M.Sc. (Occupational Hygiene))--North-West University, Potchefstroom Campus, 2011.

Made available in DSpace on 2014-06-11T19:32:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-20Bitstream added on 2014-06-13T20:23:16Z : No. of bitstreams: 1 maschio_lr_dr_sjrp.pdf: 1208225 bytes, checksum: 581a26de1a4603e41d2d07020f15f18d (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação para o Desenvolvimento da UNESP (FUNDUNESP) / Devido às crescentes expansões demográficas e industriais observadas nas últimas décadas, o meio ambiente tem recebido uma carga significativamente crescente de efluentes domésticos, industriais e agrícolas, causando impactos severos nos ecossistemas e um potencial comprometimento à saúde humana. Dentre os efluentes domésticos, podemos citar uma gama de poluentes, tais como químicos de diversas categorias, além de contaminações por agentes biológicos diversos. Já os efluentes industriais contêm poluentes orgânicos e/ou inorgânicos, dependendo da atividade industrial. Baseando-se nestes dados, este trabalho teve como objetivo investigar, por meio de ensaios biológicos com dois organismos-teste, a possível presença de contaminantes com potencial citotóxico, genotóxico e mutagênico, que são despejados ao longo do rio Preto, inclusive na Represa Municipal de São José do Rio Preto. O material biológico utilizado neste estudo constituiu-se de sementes de Allium cepa (cebola) e peixes da espécie Oreochromis niloticus (Tilápia). Coletas de águas foram realizadas, sazonalmente, nos meses de agosto de 2006 e 2007 (estação seca) e março de 2007 e 2008 (estação chuvosa), em seis pontos distintos: Ponto 1 (P1), 8 km antes do represamento; Ponto 2 (P2), 1 km antes do represamento; Ponto 3 (P3), local de despejo do esgoto; Ponto 4 (P4), margem oposta do despejo do esgoto; Ponto 5 (P5), saída do represamento; Ponto 6 (P6), 1 km após o represamento. Análises químicas foram realizadas para todas as coletas realizadas. Para a realização do estudo, 100 sementes de Allium cepa foram submetidas à germinação, em placa de Petri, em amostras de águas coletadas nos seis diferentes pontos do rio Preto, em água ultra pura (controle negativo) e em uma substância reconhecidamente aneugênica (Trifluralina - controle positivo), sempre à temperatura ambiente... / Due to increasing population and industrial expansion observed in recent decades, the environment has received a significant increased burden of domestic industrial and agricultural sewerage, which can cause severe impacts on ecosystems, and a potential damage to human health as well. A wide range of harmful pollutants can be found in domestic effluent, such as chemicals from various categories, in addition to contamination by various biological agents. On the other hand, industrial effluents contain organic and / or inorganic pollutants, depending on industrial activity. Based on these data, this study aimed to investigate, by means of biological tests with two test-organism, the possible presence of contaminants with cytotoxic, genotoxic and mutagenic potential, which are dumped along the Preto river, an important river that flows in the region of Sao Jose do Rio Preto/SP. The biological material used in this study consisted of seeds of Allium cepa (onion) and one specie of fish (Tilapia: Oreochromis niloticus). Water samples were taken seasonally in August 2006 and 2007 (dry season) and March 2007, and 2008 (rainy season), in six distinct sites: Site 1 (S1), 8 km before the damming, Site 2 (S2), 1 km before the damming, Site 3 (S3), place of sewerage discharge; Site 4 (S4), opposite margin of sewage discharge, Site 5 (S5), end of the damming; Site 6 (S6) 1 km after damming. Chemical analyses were performed for all collected samples. For the study, 100 seeds of A. cepa were submitted to germination in Petri dishes with samples water from six different sites of the Preto river, Ultra pure water (negative control), and with an aneugenic substance (Trifluralin - positive control). For most of collection points and periods studied, root meristems cells of A. cepa, exposed to water samples collected along the Preto river, showed no significant differences... (Complete abstract click electronic access below)