No description available.

No description available.

O chumbo (Pb) é um metal tóxico que se acumula no organismo e provoca diversos efeitos, afetando vários sistemas, tais como renal, gastrointestinal, reprodutor, endócrino, hepático e hematopoiético. No que se refere a estudos epigenéticos, quando comparados a outros metais, o Pb é, até o momento, o menos estudado e, portanto, muito pouco se sabe a respeito dos efeitos epigenéticos sobre a absorção, distribuição, metabolismo e excreção (ADME) do Pb e, consequentemente, sobre a toxicidade induzida pela exposição ao Pb. Assim, o objetivo do presente estudo consiste em avaliar o padrão de metilação de regiões promotoras dos genes GCLC e MT2A, quantificar a porcentagem de metilação global além das dosagens de parâmetros hepáticos e renais e associá-los à exposição ao Pb. De acordo com os guidelines STROBE e STREGA, 100 trabalhadores do sexo masculino com idades entre 18 e 67 anos participaram do trabalho. Cerca de 6,0 mL de sangue e 5,0 mL de soro foram coletados após a assinatura do Termo de Consentimento Livre e Esclarecido (TCLE). As concentrações de Pb no sangue (Pb-s) e no plasma (Pb-p) foram determinadas por espectrometria de massas com plasma indutivamente acoplado (ICP-MS); o DNA genômico foi extraído do sangue total e quantificado por kits específicos. As amostras de DNA foram tratadas com bissulfito de sódio e a quantificação da porcentagem de metilação global do DNA dos indivíduos expostos ao metal foi determinada pelo método de Elisa indireto e o sequenciamento das amostras para avaliar o padrão de metilação de regiões promotoras dos genes GCLC (Subunidade Catalítica de Glutamato-Cisteína Ligase) e MT2A (Metalotioneína 2A), pela técnica de pirosequenciamento. Também foram determinadas as atividades das enzimas transaminase glutâmica oxalacética (TGO) e pirúvica (TGP), bem como da gama-glutamil transferase (GGT) no plasma por espectrofotometria UV/Visível. Cerca de 32% dos voluntários consomem bebidas alcoólicas regularmente e 12% são fumantes. A concentração de Pb-s em média foi de 19 ± 10 ?g/dL (variando de 1,8 a 48 ?g/dL) e Pb-p 0,56 ± 0,64 ?g/dL atingindo valores de até 4,0 ?g/dL. A maioria dos indivíduos, apresentou concentrações de ureia, creatinina, TGO, TGP e GGT dentro dos valores de referência. Em relação aos marcadores epigenéticos, foi observada uma associação inversa entre concentrações de Pb-s e Pb-p, ou seja, indivíduos que apresentavam altas concentrações do metal, tinham menor % de metilação global do DNA. Em relação ao estudo das regiões promotoras dos genes MT2A e GCLC, os resultados sugerem que o Pb não altera a metilação nas ilhas CpGs localizadas nas regiões promotoras dos genes MT2A e GCLC, entretanto, foi capaz de induzir alterações no padrão de metilação global do DNA, fornecendo resultados acerca das possíveis interações entre o genoma e o metal. / Lead (Pb) is a toxic metal that accumulates in the body and induces several effects, affecting several systems, such as renal, gastrointestinal, reproductive, endocrine, hepatic and hematopoietic. Regarding epigenetic studies, when compared to other metals, Pb is the least studied and, therefore, very little is known about the epigenetic effects on absorption, distribution, metabolism and excretion (ADME) of Pb and, consequently, on the toxicity induced by the metal. Therefore, the aim of the present study was to evaluate the methylation pattern of GCLC and MT2A gene promoter regions, to quantify the percentage of global methylation in addition to hepatic and renal measurement levels and to associate them with Pb exposure. According to the STROBE and STREGA guidelines, 100 male workers between 18 and 67 years old participated in the study. About 6.0 mL of blood and 5.0 mL of serum were collected after signing the free and informed consent term (TCLE). Pb concentrations in blood (b-Pb) and plasma (p-Pb) were determined by inductively coupled plasma mass spectrometry (ICP-MS); the genomic DNA was extracted from whole blood and quantified by specific kits. The DNA samples were treated with sodium bisulfite and the quantification of the DNA global methylation percentage of workers exposed to the metal was determined by the indirect Elisa method and the sequencing of the samples to evaluate the methylation pattern of GCLC (Catalytic Subunit of Glutamate-Cysteine Ligase) and MT2A (Metallothionein 2A) by the technique of pyrosequencing. The activities of glutamic oxalacetic transaminase (TGO) and pyruvic (TGP) enzymes as well as gamma-glutamyl transferase (GGT) in plasma were also determined by UV / Visible spectrophotometry. About 32% of volunteers regularly drink alcohol and 12% are smokers. The mean b-Pb concentration was 19 ± 10 ?g / dL (ranging from 1.8 to 48 ?g / dL) and p-Pb 0.56 ± 0.64 ?g / dL reaching values up to 4.0 ?g / dL. Most of the individuals had concentrations of urea, creatinine, TGO, TGP and GGT within the reference values. In relation to the epigenetic markers, an inverse association was observed between b-Pb and p-Pb concentrations, individuals with high concentrations of the metal, had lower % of global DNA methylation. Regarding the study of the promoter regions of the MT2A and GCLC genes, the results suggest that Pb does not alter the methylation in the CpG islands located in the promoter regions of the MT2A and GCLC genes, however, it was able to induce changes in the DNA global methylation pattern, providing results on possible interactions between the genome and the metal.

Relative to other metals, little is known about lead toxicity in fish. The use of predictive models such as the biotic ligand model (BLM) has been limited, a situation that is at least partially due to the lack of understanding of lead's acute toxic mechanism and characterization of key binding sites involved in this toxicity. Using the rainbow trout as a model species, the acute toxic mechanism for lead was found to be ionoregulatory disruption. While having no apparent respiratory or acid/base effects, Pb exposure resulted in significant ionoregulatory impacts that affected Ca2+ homeostasis, as well as Na+ and Cl- balance. Active Ca2+ uptake by the gills obeyed typical Michaelis-Menten kinetics, and Pb interacted in a competitive fashion with the uptake process. Exposure to increasing waterborne Pb concentrations resulted in significant increases in Km value while Jmax showed little or no change. A slower, non-competitive interaction occurred after prolonged Pb-exposure, evidenced by a significant reduction of high-affinity Ca2+ -ATPase activity that correlated well with branchial Pb accumulation. Conversely, calcium had a protective effect against branchial Pb accumulation, this relationship being predominately competitive in nature. Voltage-independent calcium channel blockers La3+, Cd, and Zn significantly reduced gill Pb burden while the voltage-dependent, L-type calcium channel blockers, nifedipine and verapamil, did not, suggesting Pb enters fish by a similar mechanism to that of Ca2+. Stimulated stanniocalcin release by CaCl2 injection also significantly reduced branchial Pb accumulation. Based on the evidence presented in this thesis, it is apparent that acute Pb toxicity occurs by ionoregulatory disruption. It is likely that Pb shares a similar uptake pathway as that for Ca2+ and that resulting accumulation results in disruption of Ca2+ influx as well as Na+ and Cl- balance. This study has provided data essential to the characterization of key binding sites involved in Pb toxicity, and ultimately, validates the development and application of predictive models such as the BLM. / Thesis / Master of Science (MSc)

No description available.

A lack of consistent, evidence-based practices for blood lead testing of children existed in a local public health department (LHD). No known blood lead level is safe, and toxicity can result in behavioral and cognitive impairments. The purpose of this project was to develop and analyze a clinical practice guideline to establish blood lead testing procedures in the LHD to improve testing procedures and enhance future testing within the jurisdiction. The RE-AIM framework was used to address the reach, effectiveness, adoption, implementation, and maintenance of the clinical practice guideline. Five experts evaluated the guideline using the Appraisal of Guidelines for Research and Evaluation instrument. The assessment results indicated 96.4% agreement across all domains. The experts agreed unanimously to recommend adoption of the clinical practice guideline. Implementation of the guideline might advance nursing practice and patient care in the LHD through incorporation of evidence-based practices. Implementation might also lead to early identification of lead-burdened children and may provide the opportunity for treatment to mitigate cognitive and behavioral deficits related to lead toxicity, thereby improving child health and decreasing related health care costs. Engagement of the clinical practice guideline will support positive social change through the empowerment of public health nurses to provide optimal care to a population of children at risk of deleterious and long-term side effects of lead exposure.

A exposição a baixos níveis de chumbo é associada a desordens cognitivas e neurológicas em crianças. Foi descrito um aumento dos níveis de chumbo no sangue de crianças que vivem em comunidades que recebem água fluoretada. Este estudo testou se flúor a 100 ppm na forma de ácido fluorsilícico na água induziria aumento na concentração de chumbo no sangue e tecidos calcificados de ratos Wistar expostos a baixos níveis de chumbo desde a idade gestacional. Ratas foram separadas em quatro grupos: controle e grupos que receberam água que continham 100 mg/L de fluoreto (F), 30 mg/L de chumbo (Pb), ou 100 mg/L de F e 30 mg/L de Pb (F+Pb), desde uma semana antes do acasalamento até que os filhotes completassem 81 dias. Todos os animais foram pesados ao longo do experimento. Sangue e tecidos calcificados foram coletados com 81 dias para análise de chumbo, flúor e fósforo em esmalte, dentina, osso total, osso superficial e sangue. Chumbo foi determinado por ICP-MS (Inductively Coupled Plasma Mass Spectrometry). Flúor foi medido através de elétrodo íon-específico e fósforo foi determinado por reação colorimétrica. As concentrações de chumbo triplicaram no grupo F+Pb (76.7±11.0 µg/dL) em comparação ao grupo Pb (22.6±8.5 µg/dL)(p <0.001), com a mesma tendência observada em todos os tecidos calcificados analisados (p <0.001 para todas as comparações). No esmalte, as concentrações de chumbo analisadas foram 2.5 vezes mais altas no grupo F+Pb em comparação ao grupo Pb (4,369±1,353 µg/g versus 1,768±1,892 µg/g). Na dentina, a concentração de chumbo encontrada no grupo F+Pb era 8.5±2.0 µg/g versus 4.9±1.7 µg/g no grupo Pb. Na superfície óssea, a concentração de chumbo encontrada no grupo F+Pb era 3.1 vezes maior do que as determinadas no grupo Pb, com 28.0±10.6 e 9.0±3.7 µg/g nos grupos F+Pb e Pb, respectivamente. No osso total, os valores de chumbo dobraram no grupo F+Pb (14.2±2.6 µg/g) em comparação com o grupo Pb (6.8±1.7 µg/g). Os valores de chumbo estavam abaixo do limite de detecção na maioria das amostras dos grupos Controle e F. As concentrações de flúor aumentaram em ambos os grupos expostos a flúor (F e F+Pb), com diferenças estatisticamente significantes dos grupos controle e Pb, mas nenhuma diferença foi encontrada nas concentrações de F entre os grupos F e F+Pb em quaisquer dos tecidos calcificados testados. Em conclusão, este estudo mostra um aumento nas concentrações de chumbo no sangue total, esmalte, dentina, superfície óssea e osso total em ratas com 81 dias expostas ao flúor e chumbo desde o período pré-natal. / Low-level lead exposure is linked to cognitive and neurological disorders in children. An increased risk for higher blood lead levels was described for children living in communities that receive fluoridated-water. This study tested whether water fluoride would induce increases in the blood and calcified tissue lead concentrations in Wistar rats exposed to low lead levels since gestational age. Female rats were allocated in four groups: control, and 3 groups that received water containing 100 mg/L of fluoride (F), 30 mg/L of lead (Pb), or 100 mg/L of F and 30 mg/L of Pb (F+Pb). Females mated and delivered their pups receiving the same water treatment. Female pups were maintained on the same water regimen until day 81. Lead was determined by ICP-MS (Inductively Coupled Plasma Mass Spectrometry), and fluoride was measured by ion-specific electrode in the whole blood, superficial enamel, dentine, surface bane, and whole bane. Mean whole blood lead concentrations triplicated in the F+Pb group (76.7±11.0 µg/dL) in comparison to the Pb group (22.6±8.5 µg/dL)(p < 0.001), with the same trend observed in all calcified tissues analyzed (p < 0.001 for all comparisons). In the enamel, mean lead concentrations were 2.5 times higher in the F+Pb group compared with the Pb group (4,369±1,353 µg/g versus 1,768±892 µg/g). In dentine, mean lead concentration found in F+Pb group was 8.5 ±2.0 µg/g versus 4.9±1.7 µg/g in the Pb group. In the bane surface, the mean lead concentration found in the F+Pb group was 3.1 times that determined in the Pb animals, with 28.0±10.6 and 9.0±3.7 µg/g in the F+Pb and Pb groups, respectively. In whole bane, mean lead values doubled in the F+Pb group (14.2±2.6 µg/g) in comparison with the Pb group (6.8±1.7 µg/g). Lead values were below detection limit in most Control and F group samples. Fluoride concentrations were increased in both groups exposed to fluoride (F and F+Pb), with statistically significant differences from the control and Pb groups, but no differences in the F concentrations were found between the F and F+Pb group in any of the calcified tissues tested. In conclusion, this study shows a fluoride-induced increase in the concentrations of lead of whole blood, enamel, dentine, surface bane and whole bane of 81-day female rats exposed to lead since gestational age, suggesting that a biological effect not recognized so far may underlie the epidemiological association between increased blood lead levels in children living in water-fluoridated communities.

A exposição a baixos níveis de chumbo é associada a desordens cognitivas e neurológicas em crianças. Foi descrito um aumento dos níveis de chumbo no sangue de crianças que vivem em comunidades que recebem água fluoretada. Este estudo testou se flúor a 100 ppm na forma de ácido fluorsilícico na água induziria aumento na concentração de chumbo no sangue e tecidos calcificados de ratos Wistar expostos a baixos níveis de chumbo desde a idade gestacional. Ratas foram separadas em quatro grupos: controle e grupos que receberam água que continham 100 mg/L de fluoreto (F), 30 mg/L de chumbo (Pb), ou 100 mg/L de F e 30 mg/L de Pb (F+Pb), desde uma semana antes do acasalamento até que os filhotes completassem 81 dias. Todos os animais foram pesados ao longo do experimento. Sangue e tecidos calcificados foram coletados com 81 dias para análise de chumbo, flúor e fósforo em esmalte, dentina, osso total, osso superficial e sangue. Chumbo foi determinado por ICP-MS (Inductively Coupled Plasma Mass Spectrometry). Flúor foi medido através de elétrodo íon-específico e fósforo foi determinado por reação colorimétrica. As concentrações de chumbo triplicaram no grupo F+Pb (76.7±11.0 µg/dL) em comparação ao grupo Pb (22.6±8.5 µg/dL)(p <0.001), com a mesma tendência observada em todos os tecidos calcificados analisados (p <0.001 para todas as comparações). No esmalte, as concentrações de chumbo analisadas foram 2.5 vezes mais altas no grupo F+Pb em comparação ao grupo Pb (4,369±1,353 µg/g versus 1,768±1,892 µg/g). Na dentina, a concentração de chumbo encontrada no grupo F+Pb era 8.5±2.0 µg/g versus 4.9±1.7 µg/g no grupo Pb. Na superfície óssea, a concentração de chumbo encontrada no grupo F+Pb era 3.1 vezes maior do que as determinadas no grupo Pb, com 28.0±10.6 e 9.0±3.7 µg/g nos grupos F+Pb e Pb, respectivamente. No osso total, os valores de chumbo dobraram no grupo F+Pb (14.2±2.6 µg/g) em comparação com o grupo Pb (6.8±1.7 µg/g). Os valores de chumbo estavam abaixo do limite de detecção na maioria das amostras dos grupos Controle e F. As concentrações de flúor aumentaram em ambos os grupos expostos a flúor (F e F+Pb), com diferenças estatisticamente significantes dos grupos controle e Pb, mas nenhuma diferença foi encontrada nas concentrações de F entre os grupos F e F+Pb em quaisquer dos tecidos calcificados testados. Em conclusão, este estudo mostra um aumento nas concentrações de chumbo no sangue total, esmalte, dentina, superfície óssea e osso total em ratas com 81 dias expostas ao flúor e chumbo desde o período pré-natal. / Low-level lead exposure is linked to cognitive and neurological disorders in children. An increased risk for higher blood lead levels was described for children living in communities that receive fluoridated-water. This study tested whether water fluoride would induce increases in the blood and calcified tissue lead concentrations in Wistar rats exposed to low lead levels since gestational age. Female rats were allocated in four groups: control, and 3 groups that received water containing 100 mg/L of fluoride (F), 30 mg/L of lead (Pb), or 100 mg/L of F and 30 mg/L of Pb (F+Pb). Females mated and delivered their pups receiving the same water treatment. Female pups were maintained on the same water regimen until day 81. Lead was determined by ICP-MS (Inductively Coupled Plasma Mass Spectrometry), and fluoride was measured by ion-specific electrode in the whole blood, superficial enamel, dentine, surface bane, and whole bane. Mean whole blood lead concentrations triplicated in the F+Pb group (76.7±11.0 µg/dL) in comparison to the Pb group (22.6±8.5 µg/dL)(p < 0.001), with the same trend observed in all calcified tissues analyzed (p < 0.001 for all comparisons). In the enamel, mean lead concentrations were 2.5 times higher in the F+Pb group compared with the Pb group (4,369±1,353 µg/g versus 1,768±892 µg/g). In dentine, mean lead concentration found in F+Pb group was 8.5 ±2.0 µg/g versus 4.9±1.7 µg/g in the Pb group. In the bane surface, the mean lead concentration found in the F+Pb group was 3.1 times that determined in the Pb animals, with 28.0±10.6 and 9.0±3.7 µg/g in the F+Pb and Pb groups, respectively. In whole bane, mean lead values doubled in the F+Pb group (14.2±2.6 µg/g) in comparison with the Pb group (6.8±1.7 µg/g). Lead values were below detection limit in most Control and F group samples. Fluoride concentrations were increased in both groups exposed to fluoride (F and F+Pb), with statistically significant differences from the control and Pb groups, but no differences in the F concentrations were found between the F and F+Pb group in any of the calcified tissues tested. In conclusion, this study shows a fluoride-induced increase in the concentrations of lead of whole blood, enamel, dentine, surface bane and whole bane of 81-day female rats exposed to lead since gestational age, suggesting that a biological effect not recognized so far may underlie the epidemiological association between increased blood lead levels in children living in water-fluoridated communities.

Im Rahmen der Studien zur Wirkung von Bleinitrat auf die Embryonalstadien des afrikanischen Welses Clarias gariepinus wurde zunächst der Einfluß der Besamung auf den Härtungsprozess des Chorions untersucht, um die Bedeutung des gehärteten Chorions als Schutzfunktion im Hinblick auf Schadstoffeinwirkung zu klären. Das Studium der Embryonalentwicklung war erforderlich, um das Ausmaß der Änderung der Normalentwicklung unter dem Einfluß von Bleinitrat bewerten zu können. Im Rahmen der toxikologischen Untersuchungen der Wirkung des Bleinitrats auf die Embryonalstadien wurden folgende biologische Marker (Biomarker) betrachtet: Änderungen in der Entwicklung und der Schlüpfrate, morphologische und histologische Änderungen, sowie biochemische Veränderungen (Änderungen von Stoffwechsel-Enzymaktivitäten) und molekulare Veränderungen (Erfassung von DNA-Schädigungen). Die Exposition der besamten Eier mit Bleinitrat führte zu einer Verlängerung der Inkubationszeit und zu starken Mißbildungen. Der Rückgang der Häufigkeiten der Mißbildungen mit der Zeit ließ die Annahme zu, daß die mißgebildeten Embryonen starben. Im Gegensatz zu den morphologischen Mißbildungen wurden histopathologische Effekte nur bei Embryonen gefunden, die den höchsten Dosierungen (300 µg/l und 500 µg/l Bleinitrat) ausgesetzt waren. Nach dem Schlupf war das Muster der Enzymaktivitäten nach Exposition mit Bleinitrat uneinheitlich; die Aktivität von G6PDH nahm zu, die von LDH nahm ab und die von PK zeigte unregelmäßige Fluktuationen. Die Embryonalstadien zeigten signifikante Dosis-abhängige Antworten über die Zeit, da das Ausmaß der DNA-Schädigungen signifikant mit den Bleinitrat Konzentrationen anstieg. Vor dem Schlupf konnten bei den Embryonen nach Bleinitrat Exposition keine Änderungen in den Enzymaktivitäten gefunden werden und nur geringe DNA-Schädigungen, d.h die toxischen Effekte waren sehr gering. Eine Erklärung könnte die schützende Wirkung der Eihülle gegenüber Schadstoffen sein. Die gewählten Biomarker stellen sensitive Detektionsmethoden für Bleinitrat dar. So könnten sie sich als sinnvolle Bioindikatoren für Ägypten erweisen, da dort zunehmend Umweltverschmutzung mit Blei und Bleiakkumulation in Lebensmitteln zu verzeichnen ist. / In order to study the embryo-toxic effects of lead nitrate of the African catfish Clarias gariepinus, we first had to study the effect of fertilization on the hardening process of the chorion to clarify the role of the hardened chorion on the protection of the embryo from the pollutants. Also we had to study the embryonic development of C. gariepinus for providing us with a model for comparison when normal patterns of development are altered due the exposure to lead nitrate. The present toxicological work focuses on lead toxicity in different developmental stages of C. gariepinus considering different biological markers (biomarkers) comprising changes in the development and hatching rate, morphological and histological changes, biochemical changes (alteration of metabolic enzymes activity) and molecular changes (monitoring of DNA damage). Exposure of fertilized eggs to lead nitrate prolonged the incubation period and caused severe morphological malformations. Since the frequencies of the morphological malformations decreased with time, we conclude a lethal impact and selected mortality of abnormal embryos. Unlike the morphological malformation, histopathological changes were only recorded in embryos exposed to the highest dosages (300 µg/l and 500 µg/l lead nitrate). In the post-hatching stages, the patterns of the enzymes activities after lead exposure varied, G6PDH increased, LDH decreased and PK showed fluctuations. Embryonic stages revealed significant dose-related DNA damage response over time, since the degree of DNA damage increased significantly with higher lead concentrations. No specific response in the activities of the selected enzymes and low DNA damage were recorded in the pre-hatching stage after exposure to the lead nitrate doses. This means the lead nitrate had a minute toxic effect on the pre-hatched embryos. We conclude that, low susceptibility in pre-hatching stages is most probably a consequence of the chorion, which seems to protect the embryos from a range of external pollutants. The selected biomarkers were sensitive detection methods for low-level toxicity of lead nitrate. Thus, these are useful tools for biomonitoring, urgently required in Egypt with regard to increasing environmental deposition of lead and bioaccumulation in human food recently observed.