Intracellular accumulation and DNA damage caused by methylmercury in glial cells.

Mercury is widely used in industrial and extractive processes, and the improper disposal of waste or products containing this metal produces a significant impact on ecosystems, causing adverse effects on living organisms, including humans. Exposure to methylmercury, a highly toxic organic compound, causes important neurological and developmental impairments. Recently, the genotoxicity of mercurial compounds has gained prominence as one of the possible mechanisms associated with the neurological effects of mercury, mostly by disturbing the mitotic spindle and causing chromosome loss. In this sense, it is important to investigate if these compounds can also cause direct damage to DNA, such as single and double-strand breaks. Thus, the aim of this study was to investigate the cytotoxic and genotoxic potential of methylmercury in cell lines derived from neurons (B103) and glia (C6), exposed to methylmercury (MeHg) for 24 h, by analyzing cell viability, metabolic activity, and damage to DNA and chromosomes. We found that in comparison to the neuronal cell line, glial cells showed higher tolerance to MeHg, and therefore a higher LC50 and consequent higher intracellular accumulation of Hg, which led to the occurrence of several genotoxic effects, as evidenced by the presence of micronuclei, bridges, sprouts, and chromosomal aberrations.

Copy Number Alterations in Papillary Thyroid Carcinomas: Does Loss of SESN2 Have a Role in Age-related Different Prognoses?

BACKGROUND/AIM: Thyroid cancer is the only tumor in which age is an important prognostic factor. In papillary thyroid carcinomas (PTC), 45 years of age seems to be a key point that divides adult patients into two groups, with different clinical features. The aim of the study was to perform a microarray-based analysis in two groups of patients (<45 and ≥45 years old), in order to verify the occurrence of specific copy number alterations (CNAs) that could be associated to different patient behaviors associated with age. PATIENTS AND METHODS: In order to search and compare genomic alterations that may be related to age, we evaluated the occurrence of CNAs in the genome of 24 PTC samples, divided in two groups (<45 and ≥45 years old). RESULTS: We identified only one region showing a statistically significant difference between the groups (p=0.00357): a deletion of approximately 537 kps in 1p35.3., which was more frequent in patients aged 45 years or older. This is the region where, among others, the gene SESN2 is located, which is activated under oxidative stress and plays an antioxidant role, in addition to protecting the genetic material from damage generated by reactive oxygen species (ROS). CONCLUSION: This is the first time that a CNA involving the deletion of the SESN2 gene is associated with papillary thyroid carcinomas, particularly in patients aged 45 years and older, indicating that this deletion would lead to a more malignant and prominent tumoral behavior associated to a worst prognosis.

Meningiomas: An Overview of the Landscape of Copy Number Alterations in Samples from an Admixed Population.

Meningiomas are considered the most common intracranial tumors, affecting mainly women. Studies in mixed populations can be of great importance to clarify issues related to the genetic diversity of tumors and their development. Considering that data obtained from analyses of the profile of copy number alterations (CNA) have been a useful diagnostic indicator for many types of tumors and that meningiomas show a complex pattern of gains and losses in the number of copies, our objective was to analyze the CNA profile in 33 samples of meningiomas of different histological grades (WHO Grade I-III) from patients in a city located in the Amazon region of Brazil, using aCGH. We found that the female to male ratio was 3 : 1. The aCGH analysis revealed a total of 2304 CNA, with an average of 69.8 ± 57.4 per case, of which 1197 were gains (52%), 926 were losses (40.2%), 105 were amplifications (4. 5%), and 76 were deletions (3.3%). A significant relationship was observed between the type of CNA and the degree of the tumor (chi-square test: χ 2 = 65,844; p < 0.0001; contingency coefficient: C = 0.1772; p < 0.0001). Evaluating the recurrent changes in at least 50% of the samples, we observe as the most frequent losses of the segments 22q13.1-q13.2 (82%), 1p35.3 (76%), and 14q13.1-q13.2 (67%), involving all histopathological grades. The analysis of these regions showed the inclusion of genes with functions such as regulation, maintenance of cell survival, reorganization of the cytoskeleton, cell signaling, and DNA repair, among others. However, overall, the profiles observed in meningiomas of this admixed population were very similar to the ones observed in Caucasian groups. An interesting finding was a recurrent gain of 8p22 observed only in grade I meningiomas, a region which includes DLC1, a suppressor candidate gene probably implicated in the developments or progression of meningiomas, usually found deleted, when related to CNAs.

Detection and Correlation of Single and Concomitant TP53, PTEN, and CDKN2A Alterations in Gliomas.

Gliomas are the most frequent primary tumors of central nervous system and represent a heterogeneous group of tumors that originates from the glial cells. TP53, PTEN, and CDKN2A are important tumor suppressor genes that encode proteins involved in sustaining cellular homeostasis by different signaling pathways. Though genetic alterations in these genes play a significant role in tumorigenesis, few studies are available regarding the incidence and relation of concomitant TP53, PTEN, and CDKN2A alterations in gliomas. The purpose of this study was to evaluate the occurrence of mutation and deletion in these genes, through single-strand conformational polymorphism, array-comparative genomic hybridization, and fluorescence in situ hybridization techniques, in 69 gliomas samples. Molecular results demonstrated a significant higher prevalence of TP53, PTEN, and CDKN2A alterations in astrocytoma than other tumor subtypes, and heterozygous deletion was the most frequent event. In addition, a significant association was observed between TP53 and CDKN2A alterations (p = 0.0424), which tend to coexist in low grade astrocytomas (5/46 cases (10.9%)), suggesting that they are early events in development of these tumors, and PTEN and CDKN2A deletions (p = 0.0022), which occurred concomitantly in 9/50 (18%) patients, with CDKN2A changes preceding PTEN deletions, present preferably in high-grade gliomas.

Exposure to Inorganic Mercury Causes Oxidative Stress, Cell Death, and Functional Deficits in the Motor Cortex.

Mercury is a toxic metal that can be found in the environment in three different forms - elemental, organic and inorganic. Inorganic mercury has a lower liposolubility, which results in a lower organism absorption and reduced passage through the blood-brain barrier. For this reason, exposure models that use inorganic mercury in rats in order to evaluate its effects on the central nervous system are rare, especially in adult subjects. This study investigated if a chronic exposure to low doses of mercury chloride (HgCl2), an inorganic form of mercury, is capable of promoting motor alterations and neurodegenerative in the motor cortex of adult rats. Forty animals were exposed to a dose of 0.375 mg/kg/day, for 45 days. They were then submitted to motor evaluation and euthanized to collect the motor cortex. Measurement of mercury deposited in the brain parenchyma, evaluation of oxidative balance, quantification of cellular cytotoxicity and apoptosis and density of mature neurons and astrocytes of the motor cortex were performed. It was observed that chronic exposure to inorganic mercury caused a decrease in balance and fine motor coordination, formation of mercury deposits and oxidative stress verified by the increase of lipoperoxidation and nitrite concentration and a decrease of the total antioxidant capacity. In addition, we found that this model of exposure to inorganic mercury caused cell death by cytotoxicity and induction of apoptosis with a decreased number of neurons and astrocytes in the motor cortex. Our results provide evidence that exposure to inorganic mercury in low doses, even in spite of its poor ability to cross biological barriers, is still capable of inducing motor deficits, cell death by cytotoxicity and apoptosis, and oxidative stress in the motor cortex of adult rats.

Hippocampal Dysfunction Provoked by Mercury Chloride Exposure: Evaluation of Cognitive Impairment, Oxidative Stress, Tissue Injury and Nature of Cell Death.

Mercury (Hg) is a highly toxic metal, which can be found in its inorganic form in the environment. This form presents lower liposolubility and lower absorption in the body. In order to elucidate the possible toxicity of inorganic Hg in the hippocampus, we investigated the potential of low doses of mercury chloride (HgCl2) to promote hippocampal dysfunction by employing a chronic exposure model. For this, 56 rats were exposed to HgCl2 (0.375 mg/kg/day) via the oral route for 45 days. After the exposure period, the animals were submitted to the cognitive test of fear memory. The hippocampus was collected for the measurement of total Hg levels, analysis of oxidative stress, and evaluation of cytotoxicity, apoptosis, and tissue injury. It was observed that chronic exposure to inorganic Hg promotes an increase in mercury levels in this region and damage to short- and long-term memory. Furthermore, we found that this exposure model provoked oxidative stress, which led to cytotoxicity and cell death by apoptosis, affecting astrocytes and neurons in the hippocampus. Our study demonstrated that inorganic Hg, even with its low liposolubility, is able to produce deleterious effects in the central nervous system, resulting in cognitive impairment and hippocampal damage when administered for a long time at low doses in rats.

Chronic intoxication by methylmercury leads to oxidative damage and cell death in salivary glands of rats.

Methylmercury (MeHg) is one of the most toxic species of mercury, causing several systemic damages; however, its effect on the salivary glands has rarely been explored to date. This study was aimed at analyzing the mercury deposit, oxidative stress markers, and cell viability in parotid and submandibular rat salivary glands after chronic methylmercury intoxication. Herein, forty male Wistar rats (40 days old) were used in the experiment. The animals of the experimental group were intoxicated by intragastric gavage with MeHg at a dose of 0.04 mg per kg body weight per day for 35 days, whereas the control group received only corn oil, a diluent. After the period of intoxication, the glands were obtained for evaluation of total mercury deposit, cell viability, and the malondialdehyde (MDA) and the nitrite levels. Our results indicated mercury deposits in salivary glands, with a decrease in cell viability, higher levels of MDA in both glands of intoxicated animals, and a higher concentration of nitrite only in the submandibular gland of the mercury group. Thus, the intoxication by MeHg was able to generate deposits and oxidative stress in salivary glands that resulted in a decrease in cell viability in both types of glands.

Is Low Non-Lethal Concentration of Methylmercury Really Safe? A Report on Genotoxicity with Delayed Cell Proliferation.

Human exposure to relatively low levels of methylmercury is worrying, especially in terms of its genotoxicity. It is currently unknown as to whether exposure to low levels of mercury (below established limits) is safe. Genotoxicity was already shown in lymphocytes, but studies with cells of the CNS (as the main target organ) are scarce. Moreover, disturbances in the cell cycle and cellular proliferation have previously been observed in neuronal cells, but no data are presently available for glial cells. Interestingly, cells of glial origin accumulate higher concentrations of methylmercury than those of neuronal origin. Thus, the aim of this work was to analyze the possible genotoxicity and alterations in the cell cycle and cell proliferation of a glioma cell line (C6) exposed to a low, non-lethal and non-apoptotic methylmercury concentration. Biochemical (mitochondrial activity) and morphological (integrity of the membrane) assessments confirmed the absence of cell death after exposure to 3 µM methylmercury for 24 hours. Even without promoting cell death, this treatment significantly increased genotoxicity markers (DNA fragmentation, micronuclei, nucleoplasmic bridges and nuclear buds). Changes in the cell cycle profile (increased mitotic index and cell populations in the S and G2/M phases) were observed, suggesting arrest of the cycle. This delay in the cycle was followed, 24 hours after methylmercury withdrawal, by a decrease number of viable cells, reduced cellular confluence and increased doubling time of the culture. Our work demonstrates that exposure to a low sublethal concentration of MeHg considered relatively safe according to current limits promotes genotoxicity and disturbances in the proliferation of cells of glial origin with sustained consequences after methylmercury withdrawal. This fact becomes especially important, since this cellular type accumulates more methylmercury than neurons and displays a vital role protecting the CNS, especially in chronic intoxication with this heavy metal.

[Mercury exposure evaluation among Pakaanóva Indians, Amazon Region, Brazil]. / Avaliação dos níveis de exposição ao mercúrio entre índios Pakaanóva, Amazônia, Brasil.

A cross-sectional study was conducted to evaluate mercury (Hg) exposure among 910 Pakaan va Indians from the counties of Guajar Mirim and Nova Marmor , Rond nia State, Brazil. Individual hair samples were taken from the occipital region, and Hg was measured by atomic absorption spectrometry with cold vapor generation. Mean Hg in hair samples was 8.37 micro g/g (range 0.52-83.89), indicating high exposure. Young children (< 2 years old) showed a mean Hg of 10.54 micro g/g, and children from 3 to 5 years old had a mean Hg of 9.34 micro g/g. Mercury levels in women (8,91 micro g/g) were higher than in men (7.55 micro g/g), and this difference was significant (t = 3.26; p < 0.01). These results indicate the need for surveillance programs and complementary studies including the Pakaan va Indians in Rond nia State.

Avaliaçäo dos níveis de exposiçäo ao mercúrio entre índios Pakaanóva, Amazônia, Brasil / Mercury exposure among Pakaanóva Indians, Amazon Region, Brazil

Um estudo seccional foi realizado para avaliar os níveis de exposiçäo ao mercúrio (Hg) entre 910 índios Pakaanóva, residentes nos municípios de Guajará Mirim e Nova Marmoré no Estado de Rondônia, Brasil. Amostras de cabelo da regiäo occipital foram coletadas de cada participante do estudo e os teores de Hg determinados por Espectrofotometria de Absorçäo Atômica com Geraçäo de Vapor Frio. Os teores médios de Hg nas amostras de cabelo foram de 8,37µg/g (0,52-83,89µg/g), indicando exposiçäo elevada. Crianças de até 2 anos e entre 3 e 5 anos de idade apresentaram médias de 10,54µg/g e 9,34µg/g, respectivamente. Os teores médios de Hg nas mulheres (8,91µg/g ) säo mais elevados do que os valores observados nos homens (7,55µg/g), sendo esta diferença estatisticamente significante (t = 3,26; p < 0,01). Estes resultados indicam a necessidade de desenvolvimento de programas de vigilância e estudos complementares envolvendo os índios Pakaanóva no Estado de Rondônia

Distribuição dos fenótipos de grupos sangüíneos ABO e Lewis na presença de indicadores patológicos em uma população infantil da cidade de Belém do Pará / Dostribution of the phenotypes of blood groups ABO and Lewis in the presence of pathological indicators in an infantile population of Belém city

Na região Amazônica, infecções entéricas e outras parsitoses gastrointestinais têm um papel central na patogênese da desnutrição infantil e no retardo do crescimento. Foram estudadas 150 crianças da população de Belém (Pará-Brasil) a fim de identificar associações entre as infecções parasitárias intestinais e os grupos sangüíneos ABH e Lewis. Observou-se uma freqüência significativa do fenótipo secretor de substância H, particularmente nos grupos sangüíneos O/A2, associada com essas infecções e outras alterações hematológicas. Estas condições também foram consideradas como uma das causas das discordância entre os fenótipos Lewiw salivar e sangüíneo. A assoociação parece ser biologicamente aceita, desde que estes marcadores genéticos têm sido correlacionados com as doenças infecciosas através da aderência de microorganismo às células epiteliais com especificidades fenótípicas

Distribution of the ABO and Lewis blood group phenotypes in the Caninde indian reserve

Os sistemas de grupos sangüíneos ABO e Lewis foram determinados nos níveis eritrocitário e salivar em uma populaçäo indígena miscigenada da reserva Canindé. A populaçäo desta reserva está distribuída em seis vilas compostas por indivíduos originários de diferentes tribos. A distribuiçäo dos fenótipos ABO inclui 87 por cento dos indivíduos no grupo sangüíneo O e 13 por cento no grupo sangüíneo A. Em relaçäo ao fenótipo Lewis eritrocitário, foram consideradas as seguintes freqüências: 4 por cento Le (a+b-), 48 por cento Le (a-b+), 34 por centro Le (a-b-) E 14 por cento Le (a+b+). Um problema nestes achados tem sido a observaçäo de que alguns indivíduos que apresentam o fenótipo eritrocitário Le (a-b-), mostraram atividade anti-Le b em suas secreçöes salivares. A freqüência elevada do fenótipo eritrocitário (a+b+) é compatível com a média descrita para populaçöes de origem Asiática. A análise estatística com o teste G independente näo-paramétrico mostrou uma diferença näo significativa na distribuiçäo dos grupos sangüíneos ABO e Lewis nas diferentes vilas relatadas.