Characterization and in vivo toxicological evaluation of multi-walled carbon nanotubes: a low-dose repeated intratracheal administration study.

This study characterized and investigated the toxicity of two multi-walled carbon nanotubes (MWCNT) NM-401 and NM-403 at 60 and 180 µg after four repeated intratracheal instillations; follow-up times were 3, 7, 30, and 90 days after the last instillation. NM-401 was needle-like, long, and thick, while NM-403 was entangled, short, and thin. Both MWCNT types induced transient pulmonary and systemic alterations in renal function and oxidative lipid damage markers in recent times. Animals showed general toxicity in the immediate times after exposures, in addition to increased pulmonary LDH release at day 3. In further times, decreased liver and kidney relative weights were noted at higher MWCNT doses. Lung histological damages included pulmonary fibrosis, for both MWCNT types, similarly to asbestos; single liver and kidney histological alterations were present. Repeated instillations led to persistent pulmonary damage at low doses, and possibly the extrapulmonary effects may be associated with the consecutive exposures.

Children Environmentally Exposed to Agrochemicals in Rural Areas Present Changes in Oxidative Status and DNA Damage.

Rural children are exposed to several chemicals. This study evaluated the environmental co-exposure of rural children to cholinesterase inhibitor insecticides and metals/metalloids, and the resulting oxidative stress and DNA damage. Seventy-two children (5 to 16 years old) were studied at two different moments: period 1, when agrochemicals were less used, and period 2, when agrochemicals were extensively used in agriculture. Biomonitoring was performed by evaluating butyrylcholinesterase (BuChE) activity in serum; arsenic (As), chromium (Cr), lead (Pb), and nickel (Ni) levels in blood; malondialdehyde (MDA) in plasma; glutathione peroxidase (GSH-Px) and glutathione S-transferase (GST) activities in whole blood; non-protein thiol levels in erythrocytes; and micronuclei (MN) assay in exfoliated buccal cells. Cr and As levels were higher than the reference values in both periods, and Ni levels were higher than the reference values in period 2 alone. BuChE activity was inhibited in period 2 compared with period 1. In period 2, there was an increase in endogenous antioxidants and a decrease in MDA, probably demonstrating a compensatory mechanism as a response to increasing xenobiotics. Also in period 2, the MN frequency increased and BuChE and As were positively associated, suggesting co-exposure. On the other hand, in period 1, it was observed that Cr, Ni, and Pb blood levels were negatively associated with GSH-Px and GST, while MDA was positively associated with As levels. Our findings demonstrated an imbalance in endogenous antioxidants, contributing to genotoxicity and lipoperoxidation, probably in response to exposure to xenobiotics, especially carcinogenic elements (Cr, As, and Ni).

Occupational exposure to crystalline silica and peripheral biomarkers: An update.

Peripheral biomarkers are important tools for detecting occupational exposures to prevent the onset and/or progression of diseases. Studies that reveal early peripheral biomarkers are highly important to preserve the health of workers and can potentially contribute to diagnosing and/or prognosing occupational pathologies. Exposure to crystalline silica is a problem in several workplaces because it increases the risk of chronic obstructive pulmonary disease (COPD), tuberculosis, cancer, and pulmonary fibrosis, clinically defined as silicosis. Silicosis is diagnosed by chest radiography and/or lung tomography in advanced stages when there is a severe loss of lung function. Peripheral biomarkers can help in diagnosing early changes prior to silicosis and represent a highly important technical-scientific advance that is minimally invasive. This review aimed to investigate the biomarkers studied for evaluating occupational exposure to crystalline silica and to understand the recent advances in this area. Potential oxidative, inflammatory, and immunological biomarkers were reviewed, as well as routine biomarkers such as biochemical parameters. It was found that biomarkers of effect such as serum CC16 and l-selectin levels could represent promising alternatives. Additionally, studies have shown that neopterin levels in urine and serum can be used to monitor worker exposure. However, further studies are needed that include a greater number of participants, different times of exposure to crystalline silica, and a combination of silicosis patients and healthy volunteers. Evaluating the concentration of crystalline silica in occupational environments, its impact on biomarkers of effect, and alterations in lung function could contribute to revealing early health alterations in workers in a more robust manner.

Sintocalmy, a Passiflora incarnata Based Herbal, Attenuates Morphine Withdrawal in Mice.

Chronic opioid use changes brain chemistry in areas related to reward processes, memory, decision-making, and addiction. Both neurons and astrocytes are affected, ultimately leading to dependence. Passiflora incarnata L. (Passifloraceae) is the basis of frequently used herbals to manage anxiety and insomnia, with proven central nervous system depressant effects. Anti-addiction properties of P. incarnata have been reported. The aim of this study was to investigate the effect of a commercial extract of Passiflora incarnata (Sintocalmy®, Aché Laboratory) in the naloxone-induced jumping mice model of morphine withdrawal. In addition, glial fibrillary acidic protein (GFAP) and S100 calcium-binding protein B (S100B) levels were assessed in the frontal cortex and hippocampus, and DNA damage was verified on blood cells. In order to improve solubilization a Sintocalmy methanol extract (SME) was used. SME is mainly composed by flavonoids isovitexin and vitexin. The effects of SME 50, 100 and 200 mg/kg (i.p.) were evaluated in the naloxone-induced withdrawal syndrome in mice. SME 50 and SME 100 mg/kg decreased naloxone-induced jumping in morphine-dependent mice without reducing locomotor activity. No alterations were found in GFAP levels, however SME 50 mg/kg prevented the S100B increase in the frontal cortex and DNA damage. This study shows anti-addiction effects for a commercial standardized extract of P. incarnata and suggests the relevance of proper clinical assessment.

Cellular response to chemicals present in air pollution in occupationally exposed workers and its potential cancer susceptibility.

The study aimed to evaluate the potential effects of the chronic exposure to chemical agents from air pollution on phenotypic and genotypic expressions of peripheral biomarkers and tumor-related proteins in mononuclear cells. This study evaluates 85 taxi drivers (outdoor workers) and 55 non-occupationally exposed persons (NOE) to air pollution (indoor workers). The biomarkers were urinary 1-hydroxypyrene (1-OHP), for organic agents, and blood As and Ni, for inorganic agents. Oxidative stress biomarkers; protein expression of ICAM-1 (CD54), ß2-integrin, L-selectin (CD62-L), and MCP1; gene expression of ICAM-1, p53 and CD26 were performed. Urinary 1-OHP and blood As and Ni levels were increased in taxi drivers and were associated with inflammatory and oxidative stress biomarkers. These exposure biomarkers were also associated with each other, suggesting a common source of exposure. The gene expression of p53, CD26 and ICAM-1 were decreased in taxi drivers and were strongly associated between them, indicating a commom regulation point. The antioxidant non-protein thiols and lycopene were negatively associated with inflammatory biomarkers, maybe regulating the immune-response. We demonstrated, for the first time, that in occupational exposure to air pollution chemicals, oxidative and inflammatory processes are involved in the immune-regulatory process, and indirectly contribute to suppressing the p53 and CD26 expressions, increasing the risk of cancer development. On the other hand, antioxidants could contribute to improving the immune-regulation, but more studies are needed.

Biochemical, hematological and immunological parameters and relationship with occupational exposure to pesticides and metals.

This study aimed to evaluate biomarkers of exposure to cholinesterase inhibitors insecticides (AChE and BuChE activities) and metals (As, Cd, Cr, Mn, Ni, and Pb blood levels) and their associations with biochemical, hematological, and immunological parameters in farmers from Southern Brazil. One hundred and sixteen individuals were divided into two groups: 62 farmers (exposed group) and 54 subjects non-occupationally exposed (NOE) to agrochemicals. Erythrocyte (AChE) and serum (BuChE) cholinesterases activities were significantly reduced as well as blood Cd and Pb levels were increased in farmers when compared to NOE group (p < 0.05). Farmers presented increased glucose and urea levels compared to NOE group, which were inversely associated with AChE and positively correlated with Cd (p < 0.05), respectively. In addition, Cd was inversely associated with the hematological cells counts, which were significantly reduced in farmers (p < 0.05). C3 complement was higher in farmers and was positively associated with blood Pb (p < 0.05). Surface protein expression analysis revealed a downregulation of LFA-1 and ICAM-1 in farmers. Inverse associations were found between LFA-1 and blood As, Cr, and Ni levels (p < 0.05). Taken together, our results pointed to a relationship between agrochemicals and metals exposure and biochemical, hematological, and immunological disorders that can lead to several chronic conditions.

Piperazine designer drugs elicit toxicity in the alternative in vivo model Caenorhabditis elegans.

Piperazine designer drugs are a group of synthetic drugs of abuse that have appeared on the illicit market since the second half of the 1990s. The most common derivatives are 1-benzylpiperazine (BZP), 1-(4-methoxyphenyl)piperazine (MeOPP) and 1-(3,4-methylenedioxybenzyl)piperazine (MDBP). They can be consumed as capsules, tablets, but also in powder or liquid forms. Generally, although less potent than amphetamines, piperazines have dopaminergic and serotonergic activities. The aim of this work was to evaluate the toxic effects of BZP, MeOPP and MDBP using Caenorhabditis elegans as in vivo model for acute toxicity, development, reproduction and behavior testing. The LC50 for BZP, MeOPP and MDBP were 52.21, 5.72 and 1.22 mm, respectively. All concentrations induced a significant decrease in the body surface of the worms, indicating developmental alterations, and decrease in the brood size. Worms exposed to piperazine designer drugs also presented a decrease in locomotor activity and mechanical sensitivity, suggesting the possible dysfunction of the nervous system. Neuronal damage was confirmed through the decrease in fluorescence of BY200 strains, indicating loss of dopaminergic transporters. In conclusion, we suggest that piperazine designer drugs lead to neuronal damage, which might be the underlying cause of the altered behavior observed in humans.

Cigarette smoking and antioxidant defences in packed red blood cells prior to storage.

BACKGROUND: Red blood cells from smoking donors can have more lesions from oxidative stress, decreasing the benefits of blood transfusion. We aimed to explore the effect of cigarette smoking on the oxidative status of packed red blood cells (PRBCs) prior to storage. MATERIALS AND METHODS: We compared serum vitamin C, plasmatic malondialdehyde (MDA), and non-protein thiol groups (GSH) levels in PRBCs, as well glutathione peroxidase (GPx) and glutathione s-transferase (GST) activity in PRBCs from smoking (n=36) and non-smoking (n=36) donors. We also correlated urinary cotinine levels with these parameters. RESULTS: Cigarette smoking was associated with decreased serum levels of vitamin C and GPx, and increased GST activity in PRBCs. We found negative correlations between cotinine, GPx activity and vitamin C levels, and a positive correlation between cotinine and GST activity. DISCUSSION: Cigarette smoking changed antioxidant defences of PRBCs prior to storage and these parameters are correlated with cotinine levels. Increased RBC antioxidants such as GST may reflect an exposure to oxidants during erythropoiesis. Because of the inability of mature RBCs to resynthesise antioxidants, PRBCs from smokers may have higher risk of storage lesions than those from non-smoker donors.

In vitro cardiotoxicity evaluation of graphene oxide.

Graphene is a two-dimensional (2D) monolayer of carbon atoms, tightly packed, forming a honey comb crystal lattice, with physical, chemical, and mechanical properties greatly used for energy storage, electrochemical devices, and in nanomedicine. Many studies showed that nanomaterials have side-effects on health. At present, there is a lack of information regarding graphene and its derivatives including their cardiotoxic properties. The aim of the present study was to evaluate the toxicity of nano-graphene oxide (nano-GO) in the rat cardiomyoblast cell line H9c2 and the involvement of oxidative processes. The cell viability was evaluated with the fluorescein diacetate (FDA)/propidium iodide (PI) and in the trypan blue exclusion assay, furthermore mitochondrial membrane potential and production of free radicals were measured. Genotoxicity was evaluated in comet assay and low molecular weight DNA experiment. Reduction of cell viability with 20, 40, 60, 80, and 100 µg/mL nano-GO was observed after 24 h incubation. Besides, nano-GO induced a mitochondrial hyperpolarization and a significant increase of free radicals production in the same concentrations. DNA breaks were observed at 40, 60, 80, and 100 µg/mL. This DNA damage was accompanied by a significant increase in LMW DNA only at 40 µg/mL. In conclusion, the nano-GO caused cardiotoxicity in our in vitro model, with mitochondrial disturbances, generation of reactive species and interactions with DNA, indicating the importance of the further evaluation of the safety of nanomaterials.

Inflammatory and oxidative stress biomarkers at protein and molecular levels in workers occupationally exposed to crystalline silica.

Workers chronically exposed to respirable crystalline silica (CS) are susceptible to adverse health effects like silicosis and lung cancer. This study aimed to investigate potential early peripheral biomarkers of inflammation and oxidative stress in miners. The subjects enrolled in this study were occupationally unexposed workers (OUW, n = 29) and workers exposed to crystalline silica (WECS), composed by miners, which were divided into two subgroups: workers without silicosis (WECS I, n = 39) and workers diagnosed with silicosis, retired from work (WECS II, n = 42). The following biomarkers were evaluated: gene expression of L-selectin, CXCL2, CXCL8 (IL-8), HO-1, and p53; malondialdehyde (MDA) plasma levels and non-protein thiol levels in erythrocytes. Additionally, protein expression of L-selectin was evaluated to confirm our previous findings. The results demonstrated that gene expression of L-selectin was decreased in the WECS I group when compared to the OUW group (p < 0.05). Regarding gene expression of CXCL2, CXCL8 (IL-8), HO-1, and p53, significant fold change decreases were observed in workers exposed to CS in relation to unexposed workers (p < 0.05). The results of L-selectin protein expression in lymphocyte surface corroborated with our previous findings; thus, significant downregulation in the WECS groups was observed compared to OUW group (p < 0.05). The MDA was negatively associated with the gene expression of CXCL-2, CXCL8 (IL-8), and p53 (p < 0.05). The participants with silicosis (WECS II) presented significant increased non-protein thiol levels in relation to other groups (p < 0.05). Taken together, our findings may contribute to help the knowledge about the complex mechanisms involved in the silicosis pathogenesis and in the risk of lung cancer development in workers chronically exposed to respirable CS.

Exposure to environment chemicals and its possible role in endocrine disruption of children from a rural area.

Endocrine disrupting chemicals (EDCs), including pesticides and metals, are present in rural areas, endangering the health of exposed populations. This work aimed to investigate the possible association between the exposure to these xenobiotics and thyroid dysfunction in children living in a rural community of Southern Brazil. Fifty-four children aged 5-16 years participated in this study. Peripheral biomarker evaluations were performed in periods of low and high exposure to pesticides. Thyroid ultrasonography was evaluated in the high exposure period. Blood levels of chromium (Cr), manganese (Mn), mercury (Hg), and lead (Pb), as well as hair Pb levels were positively correlated with thyroid stimulating hormone (TSH) concentrations and negatively associated with free thyroxine (fT4) levels in the low exposure period. Prolactin was positively associated with hair Mn in both periods. In the ultrasound tests, the majority of children presented a normal echogenicity of thyroid. Glucose was inversely associated with the biomarker of exposure to cholinesterase inhibitor insecticides, butyrylcholinesterase (BuChE). Lipid profile was above the recommended levels in both periods. In summary, our results show that children environmentally exposed to a mixture of xenobiotics in an agricultural community may have health impairments, especially on thyroid function, dyslipidemia, and glucose homeostasis disruption.

Inflammatory and oxidative stress parameters as potential early biomarkers for silicosis.

Workers involved in mining activities are exposed to crystalline silica, which leads to constant pulmonary inflammatory reactions and severe oxidative damage, resulting in silicosis. In this work, we aimed to evaluate inflammatory and oxidative stress parameters as potential early biomarkers of effect to assess crystalline silica toxicity in workers who had occupational exposure during mining. We enrolled 38 workers exposed to crystalline silica (WECS), 24 individuals with silicosis (IWS), and 30 occupationally unexposed workers (OUW), a total of 92 participants. The WECS were divided into 2 groups, according to the time of exposure: 19 workers with 1-15 years of occupational exposure (WECS I) and 19 workers with >16 years of occupational exposure (WECS II). The inflammatory parameters assessed were L-selectin, ß-2 integrin, and intercellular adhesion molecule-1 (ICAM-1) surface protein expression in lymphocytes and monocytes, complement C3 and C4, high sensitivity C-reactive protein (hsCRP), and adenosine deaminase (ADA) in serum. Plasma levels of malondialdehyde (MDA) and serum levels of vitamin C were determined as biomarkers of oxidative stress. Biochemical and hematological parameters were also investigated. L-selectin surface protein expression was significantly decreased in the WECS II group (p < 0.05), indicating the importance of this immune system component as a potential marker of crystalline-silica-induced toxicity. The MDA levels were significantly increased in the WECS I, WECS II, and IWS groups compared to the OUW group (p < 0.05). Vitamin C levels were decreased, while C3, hsCRP, ADA, and aspartate aminotransferase (AST) levels were increased in the IWS group compared to the OUW group (p < 0.05). Glucose and urea levels were significantly higher in the WECS I, II, and IWS groups compared to the OUW group (p < 0.05). Negative partial association was found between L-selectin and time of exposure (p < 0.001), supporting the relevance of this biomarker evaluation in long-term exposure to crystalline silica. Significant associations were also observed among inflammatory and oxidative stress biomarkers. Therefore, our results demonstrated the relevance of L-selectin as a potential peripheral biomarker for monitoring crystalline silica-induced toxicity in miners after chronic exposure, before silicosis has developed. However, more studies are necessary for better understanding of the use L-selectin as an early biomarker in exposed workers.

Itinerário terapêutico de mulheres com câncer de mama no sul do Brasil / Therapeutic itinerary of women with breast cancer breast in the south of Brazil

Objetivo: identificar o itinerário terapêutico de um grupo de mulheres com diagnóstico de câncer de mama. Método: estudo transversal, descritivo, com 30 pacientes usuárias do Sistema Único de Saúde (SUS) em tratamento de câncer de mama. Resultados: a idade média no momento do diagnóstico foi de 51,2 anos. O sintoma mais frequente foi o nódulo no seio (53,34%). A mastectomiafoi realizada por 100% das mulheres, 80% fizeram quimioterapia, 60% radioterapia e 100% utilizavam o Anastrazol ou Tamoxifeno. Das mulheres, 50% utilizaram o SUS para diagnóstico, 76,67% para tratamento cirúrgico e 66,67% para pagamento da quimioterapia e radioterapia. Conclusão: o itinerário percorrido para diagnóstico foi, a partir da presença de nódulos, a realização de biópsia; para tratamento, mastectomia seguida de tratamento quimioterápico, radioterápico e medicamentoso. Com relação ao plano de saúde,metade das mulheres utilizou o SUS para diagnóstico. A maioria utilizou SUS para tratamento cirúrgico e quimioterápico.

Objective: to identify the therapeutic itinerary of a group of women diagnosed with breast cancer. Method: cross-sectional,descriptive study with 30 patients, users of the Unified Health System (SUS), in treatment of breast cancer. Results: the mean age atdiagnosis was 51.2 years. The most common symptom was the nodule in the breast (53.34%). The mastectomy was performed by 100%of the women, 80% did chemotherapy, 60% radiotherapy, and 100% used Anastrazol or Tamoxifen. Of the women, 50% used SUSfor diagnosis, 76.67% for surgical treatment and 66.67% for payment of chemotherapy and radiotherapy. Conclusion: the route takenfor diagnosis was, from the presence of nodules, the biopsy; for treatment, mastectomy followed by chemotherapeutic, radiotherapeutic and drug treatment. Regarding the health plan, half of the women used SUS for diagnosis. Most of them used SUS for surgical and chemotherapeutic treatment.

Objetivo: identificar el itinerario terapéutico de un grupo de mujeres con diagnóstico de cáncer de mama. Método: estudio transversal, descriptivo, con 30 pacientes usuarias del Sistema Único de Salud (SUS) en tratamiento de cáncer de mama. Resultados:la edad media en el momento del diagnóstico fue de 51,2 años. El síntoma más frecuente fue el nódulo en el seno (53,34%). Lamastectomía fue realizada por el 100% de las mujeres, 80% hicieron quimioterapia, 60% radioterapia y 100% utilizaban el Anastrazolo Tamoxifeno. De las mujeres, 50% utilizaron el SUS para diagnóstico, 76,67% para tratamiento quirúrgico y 66,67% para pago de laquimioterapia y radioterapia. Conclusión: el itinerario recorrido para diagnóstico fue, a partir de la presencia de nódulos, la realización debiopsia; para el tratamiento, mastectomía seguida de tratamiento quimioterápico, radioterápico y medicamentoso. Con respecto al plande salud, la mitad de las mujeres utilizó el SUS para diagnóstico. La mayoría utilizó SUS para tratamiento quirúrgico y quimioterápico.

Análise dos marcadores imuno-histoquímicos associados com câncer de mama em mulheres na Região das Missões, Rio Grande do Sul, Brasil / Analysis of immunohistochemical markers associated with breast cancer in women in the Region of Missions, Rio Grande do Sul, Brazil

Objetivos: Analisar a distribuição dos subtipos moleculares de câncer de mama e correlacionar esses subtipos com o perfil etário e histológico. Métodos: Trata-se de um estudo observacional, descritivo e retrospectivo. Foram incluídas 110 mulheres com diagnóstico histológico de câncer de mama e análise imuno-histoquímica no período de janeiro de 2013 a dezembro de 2015. Com base nos laudos imuno-histoquímicos, foram definidos quatro subtipos moleculares: luminal A, luminal B, HER-2+ e triplo negativo. Foi realizada análise estatística para correlacionar subtipos moleculares e tipo histológico e idade. Resultados: A idade média das participantes do estudo foi de 56,9 anos (DP=15,15), apresentando pico na faixa etária acima dos 50 anos. O tipo histológico mais frequente (66,4%) foi o carcinoma ductal invasivo (CDI). Quanto à classifica- ção molecular, o luminal B foi o mais frequente, representando 43,6% dos casos. Conclusão: Com base nos marcadores moleculares, os carcinomas da mama foram classificados em quatro subtipos e não apresentaram diferença significativa em relação ao tipo histológico nem à idade.

Objectives: To analyze the molecular subtype's breast cancer distribution and correlate them with age and histological profile. Methods: An observational, descriptive and retrospective study. One hundred and ten women with histological diagnosis of breast cancer and immunohistochemistry analysis, from January 2013 to December 2015, were included. From the reports, four molecular subtypes were defined: luminal A, luminal B, HER-2+ and triple negative. Correlations between molecular subtypes and histologic type and age were evaluated statistically. Results: The average age of the participants of the investigation was 56.9 years (SD=15.15), with a peak in the age group over 50 years. The most common histological type (66.4%) was invasive ductal carcinoma (IDC). The luminal B was the most frequent molecular subtype, representing 43.6% of cases. Conclusions: Based on molecular markers, the breast carcinomas were classified into four subtypes and showed neither significant difference regarding histologic type nor age.