RESUMO
OBJECTIVE: This study evaluated the influence of smoking on the oral cells genotoxicity before and after at-home bleaching using 22% carbamide peroxide (CP). MATERIALS AND METHODS: This is a prospective observational analytics cohort study which evaluated nonsmokers (NS; n = 24) and smokers (S; n = 16) patients. At-home bleaching was performed using 22% CP gel in individual trays for 1 h per day for 14 days in both groups. Scrapped cells from marginal gums were collected before the bleaching treatment (D0-baseline) and 1 day (D1), 15 days (D15), and 1 month (D30) after its finishing. Cells were stained with Giemsa 10%, and the micronucleus (MN) and metanuclear alterations (MA) were counted by a trained operator in 1000 cells per patient. The collections and data analysis occurred blindly. Data was analyzed by Kruskal-Wallis, Dunn, and Mann-Whitney test (α = 0.05). RESULTS: MN frequency was not influenced by smoking or bleaching. An increase of MA was observed between D0 and D30 for both groups (p < 0.001); however, no statistical difference was found between NS and S (p > 0.05) in the evaluation times. CONCLUSION: Smoking associated with 22% carbamide peroxide gel for at-home bleaching does not show genotoxic potential analyzed by the MN counts. However, a significant increase of MA was found for smokers and nonsmokers. CLINICAL RELEVANCE: Despite of the increase in MA, smoking associated with 22% CP peroxide at-home bleaching showed no important genotoxic potential (MN) for oral cells. Therefore, at-home bleaching treatment is safe for nonsmokers and smokers even with a high carbamide peroxide concentration of 22%.
Assuntos
Clareadores Dentários , Clareamento Dental , Peróxido de Carbamida , Estudos de Coortes , Dano ao DNA , Humanos , Peróxido de Hidrogênio , Peróxidos/toxicidade , Fumar/efeitos adversos , Clareamento Dental/efeitos adversos , Clareadores Dentários/toxicidade , UreiaRESUMO
Gap junctions in liver, as in other organs, play a critical role in tissue homeostasis. Inherently, these cellular constituents are major targets for systemic toxicity and diseases, including cancer. This review provides an overview of chemicals that compromise liver gap junctions, in particular biological toxins, organic solvents, pesticides, pharmaceuticals, peroxides, metals and phthalates. The focus in this review is placed upon the mechanistic scenarios that underlie these adverse effects. Further, the potential use of gap junctional activity as an in vitro biomarker to identify non-genotoxic hepatocarcinogenic chemicals is discussed.
Assuntos
Comunicação Celular/efeitos dos fármacos , Junções Comunicantes/efeitos dos fármacos , Fígado/efeitos dos fármacos , Animais , Conexinas/biossíntese , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Humanos , Fígado/metabolismo , Metais/toxicidade , Peróxidos/toxicidade , Praguicidas/toxicidade , Ácidos Ftálicos/toxicidade , Medição de Risco , Solventes/toxicidade , Toxinas Biológicas/toxicidadeRESUMO
RESEARCH QUESTION: Can culture conditions influence the sensitivity of a Mouse Embryo Assay and its potential to detect peroxide-related toxicity in mineral oil samples? DESIGN: Protein type and concentration, embryo density and culture dish design were selected as the variables in the culture system with the potential to influence the assay's sensitivity. Fresh 1-cell mouse embryos were cultured under mineral oil samples with known peroxide concentrations. Protein type (human serum albumin [HSA]â¯+â¯α/ß-Globulins versus HSA versus bovine serum albumin [BSA]), concentration (5 mg/ml versus 0.5 mg/ml), embryo density (25 versus 3 µl/embryo) and culture dish (Petri versus micro-well dish) were adjusted to define the culture conditions with the highest sensitivity. RESULTS: High concentrations of peroxides can be easily detected by current quality control standards. However, for oil samples with a lower concentration of peroxides, supplementing the culture medium with 5 mg/ml of HSAâ¯+â¯alpha/beta-globulins or with HSA resulted in an increased detection of embryo toxicity compared with when BSA was used as the protein supplement. The sensitivity of the assay was greatly reduced when embryos were cultured in groups and when certain micro-well dishes were used. CONCLUSIONS: Current quality control protocols may not be sensitive enough to identify low concentrations of peroxides, which, if undetected, can increase over time and become potentially harmful during gamete and embryo culture. The different parameters established in this study allow the sensitivity of the Mouse Embryo Assays to be optimized to specifically detect peroxides in mineral oil samples prior to their release into the market and their broad use in human IVF.
Assuntos
Bioensaio , Técnicas de Cultura Embrionária/métodos , Embrião de Mamíferos/citologia , Camundongos/embriologia , Óleo Mineral/química , Peróxidos/isolamento & purificação , Animais , Bioensaio/métodos , Bioensaio/normas , Células Cultivadas , Meios de Cultura/química , Meios de Cultura/farmacologia , Contaminação de Medicamentos , Técnicas de Cultura Embrionária/normas , Desenvolvimento Embrionário/efeitos dos fármacos , Feminino , Fertilização in vitro/métodos , Fertilização in vitro/normas , Masculino , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Óleo Mineral/farmacologia , Peróxidos/toxicidade , Proteínas/fisiologia , Controle de Qualidade , Testes de Toxicidade/métodos , Testes de Toxicidade/normasRESUMO
OBJECTIVES: The aim of this study was to evaluate the cytotoxicity and the influence of bleaching agents on immunologically cell surface antigens of murine macrophages in vitro. MATERIALS AND METHODS: RAW 264.7 cells were exposed to bleaching gel extracts (40% hydrogen peroxide or 20% carbamide peroxide) and different H2O2 concentrations after 1 and 24-h exposure periods and 1-h exposure and 23-h recovery. Tests were performed with and without N-acetyl cysteine (NAC) and buthionine sulfoximine (BSO). Cell viability was determined by MTT assay. The expression of surface markers CD14, CD40, and CD54 with and without LPS stimulation was detected by flow cytometry, while the production of TNF-α was measured by ELISA. Statistical analysis was performed using the Mann-Whitney U test (α = 0.05). RESULTS: Extracts of bleaching agents were cytotoxic for cells after a 1-h exposure; cells could not recover after 24 h. This effect can be mitigated by the antioxidant NAC and increased by BSO, an inhibitor of glutathione (GSH) synthesis. LPS stimulated expression of all surface markers and TNF-α production. Exposure to bleaching agent extracts and H2O2 leads to a reduction of TNF-α, CD14, and CD40 expression, while the expression of CD54 was upregulated at non-cytotoxic concentrations. Whereas NAC reduced this effect, it was increased in the presence of BSO. CONCLUSIONS: Extracts of bleaching agents were irreversibly cytotoxic to macrophages after a 1-h exposure. Only the expression of CD54 was upregulated. The reactions are mediated by the non-enzymatic antioxidant GSH. CLINICAL RELEVANCE: The addition of an antioxidant can downregulate unfavorable effects of dental bleaching.
Assuntos
Antígenos de Superfície/efeitos dos fármacos , Clareadores/toxicidade , Peróxido de Hidrogênio/toxicidade , Peróxidos/toxicidade , Ureia/análogos & derivados , Acetilcisteína/farmacologia , Animais , Antígenos de Superfície/imunologia , Butionina Sulfoximina/farmacologia , Peróxido de Carbamida , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Macrófagos/efeitos dos fármacos , Camundongos , Clareamento Dental , Fator de Necrose Tumoral alfa/imunologia , Ureia/toxicidadeRESUMO
INTRODUCTION: Mesenchymal stem cells (MSCs) have been publicized to ameliorate kidney injury both in vitro and in vivo. However, very less is known if MSCs can be differentiated towards renal lineages and their further application potential in kidney injuries. MATERIALS AND METHODS: The present study developed a conditioning system of growth factors fibroblast growth factor 2, transforming growth factor-ß2, and leukemia inhibitory factor for in vitro differentiation of MSCs isolated from different sources towards nephrogenic lineage. Less invasively isolated adipose-derived MSCs were also compared to bone marrow-derived MSCs for their differentiation potential to induce renal cell. Differentiated MSCs were further evaluated for their resistance to oxidative stress induced by oxygen peroxide. RESULTS: A combination of growth factors successfully induced differentiation of MSCs. Both types of differentiated cells showed significant expression of pronephrogenic markers (Wnt4, Wt1, and Pax2) and renal epithelial markers (Ecad and ZO1). In contrast, expression of mesenchymal stem cells marker Oct4 and Vim were downregulated. Furthermore, differentiated adipose-derived MSCs and bone marrow-derived MSCs showed enhanced and comparable resistance to oxygen peroxide-induced oxidative stress. CONCLUSIONS: Adipose-derived MSC provides a promising alternative to bone marrow-derived MSC as a source of autologous stem cells in human kidney injuries. In addition, differentiated MSCs with further in vivo investigations may serve as a cell source for tissue engineering or cell therapy in different renal ailments.
Assuntos
Tecido Adiposo/citologia , Diferenciação Celular , Linhagem da Célula , Rim/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Peróxidos/toxicidade , Animais , Diferenciação Celular/genética , Linhagem Celular , Linhagem da Célula/genética , Senescência Celular , Meios de Cultura/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/patologia , Fenótipo , Ratos Sprague-DawleyRESUMO
Investigation of the Australian rainforest plant Croton insularis led to isolation of the first casbane hydroperoxide diterpenes EBC-304 and EBC-320. Extensive DFT and electronic circular dichroism (ECD) calculations in combination with 2D NMR spectroscopy determined the absolute configurations. EBC-304 and EBC-320 both display significant cytotoxicity.
Assuntos
Croton/química , Diterpenos/química , Peróxidos/química , Austrália , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Dicroísmo Circular , Croton/metabolismo , Diterpenos/isolamento & purificação , Diterpenos/toxicidade , Humanos , Espectroscopia de Ressonância Magnética , Conformação Molecular , Peróxidos/isolamento & purificação , Peróxidos/toxicidade , Floresta ÚmidaRESUMO
ABSTRACT Tooth bleaching is a technique of choice to obtain a harmonious smile, but bleaching agents may damage the dental pulp. Objective: This study evaluated the inflammatory responses of human dental pulp after the use of two bleaching techniques. Material and Methods: Pulp samples were collected from human third molars extracted for orthodontic reasons and divided into three groups: control - no tooth bleaching (CG) (n=7); at-home bleaching with 15% carbamide peroxide (AH) (n = 10), and in-office bleaching with 38% hydrogen peroxide (IO) (n=12). Pulps were removed and stained with hematoxylin-eosin for microscopic analysis of inflammation intensity, collagen degradation, and pulp tissue organization. Immunohistochemistry was used to detect mast cells (tryptase+), blood vessels (CD31+), and macrophages (CD68+). Chi-square, Kruskal-Wallis, and Mann Whitney tests were used for statistical analysis. The level of significance was set at p<.05. Results: The inflammation intensity and the number of macrophages were significantly greater in IO than in AH and CG (p<0.05). The results of CD31+ (blood vessels per mm2) were similar in CG (61.39±20.03), AH (52.29±27.62), and IO (57.43±8.69) groups (p>0.05). No mast cells were found in the pulp samples analyzed. Conclusion: In-office bleaching with 38% hydrogen peroxide resulted in more intense inflammation, higher macrophages migration, and greater pulp damage then at-home bleaching with 15% carbamide peroxide, however, these bleaching techniques did not induce migration of mast cells and increased the number of blood vessels.
Assuntos
Humanos , Pulpite/induzido quimicamente , Clareamento Dental/efeitos adversos , Polpa Dentária/efeitos dos fármacos , Clareadores Dentários/toxicidade , Peróxidos/toxicidade , Pulpite/patologia , Fatores de Tempo , Clareamento Dental/métodos , Ureia/análogos & derivados , Ureia/toxicidade , Vasos Sanguíneos/efeitos dos fármacos , Vasos Sanguíneos/patologia , Imuno-Histoquímica , Antígenos de Diferenciação Mielomonocítica , Distribuição Aleatória , Antígenos CD , Contagem de Células , Colágeno/efeitos dos fármacos , Estatísticas não Paramétricas , Molécula-1 de Adesão Celular Endotelial a Plaquetas , Polpa Dentária/patologia , Peróxido de Hidrogênio/toxicidadeRESUMO
To replace animal testing and improve the prediction of skin sensitization, significant attention has been directed to the use of alternative methods. The direct peptide reactivity assay (DPRA), the regulatory agencies' approved alternative in chemico method, has been applied for understanding the sensitization capacity of activated ascaridole. Ascaridole, the oxidative metabolite of α-terpinene, is considered to be one of the components responsible for the contact allergy associated with essential oils derived from Chenopodium and Melaleuca species. The recently developed high-throughput screening based on the dansyl cysteamine (HTS-DCYA) method was applied to understand the reported enhanced reactivity of activated ascaridole and possibly to identify the resulting elusive radical or other reactive species. For the first time, a substituted cyclohexenone was identified as a potential electrophilic intermediate resulting in higher depletion of nucleophilic DCYA, along with several nonreactive byproducts of ascaridole via a radical degradation mechanism. Formation of electrophilic species via radical degradation is one of the possible pathways should be considered for the peptide reactivity of in aged tea tree oil or oils rich in terpinenes along with commonly believed reactants, allylic-epoxides and allylic-peroxides.
Assuntos
Dermatite Alérgica de Contato , Monoterpenos/química , Monoterpenos/toxicidade , Peróxidos/química , Peróxidos/toxicidade , Pele/efeitos dos fármacos , Pele/imunologia , Animais , Monoterpenos Cicloexânicos , Cicloexanonas/química , Humanos , Estrutura Molecular , Monoterpenos/metabolismo , Peróxidos/metabolismoRESUMO
Potential biomedicinal applications of graphene oxide (GO), for example, as a carrier of biomolecules or a reagent for photothermal therapy and biosensing, are limited by its cytotoxicity and mutagenicity. It is believed that these properties are at least partially caused by GO-induced oxidative stress in cells. However, it is not known which chemical fragments of GO are responsible for this unfavorable effect. We generated four GOs containing variable redox-active groups on the surface, including Mn(2+), C-centered radicals, and endoperoxides (EPs). A comparison of the abilities of these materials to generate reactive oxygen species in human cervical cancer cells revealed that EPs play a crucial role in GO-induced oxidative stress. These data could be applied to the rational design of biocompatible nontoxic GOs for biomedical applications.
Assuntos
Grafite/química , Grafite/toxicidade , Óxidos/química , Óxidos/toxicidade , Peróxidos/toxicidade , Neoplasias do Colo do Útero/metabolismo , Materiais Biocompatíveis/química , Materiais Biocompatíveis/metabolismo , Materiais Biocompatíveis/toxicidade , Linhagem Celular Tumoral , Desenho de Fármacos , Feminino , Grafite/metabolismo , Humanos , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Óxidos/metabolismo , Peróxidos/química , Peróxidos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Neoplasias do Colo do Útero/patologiaRESUMO
O clareamento dental é um dos tratamentos mais realizados nos consultórios odontológicos a fim de melhorar a aparência do sorriso. O procedimento consiste na aplicação de um gel clareador, a base de peróxido de carbamida ou de hidrogênio, sobre os dentes a serem clareados. A sensibilidade dentária é o efeito adverso mais frequentemente relatado no clareamento dentário e é a principal causa de desmotivação dos pacientes. O mecanismo pelo qual se produz a sensibilidade após clareamento dentário ainda não foi completamente elucidado; no entanto, parece estar associado à rápida difusão dos agentes clareadores através do esmalte e dentina que, devido ao seu grau de citotoxicidade, podem agir agredindo as células pulpares, causando sensibilidade. O objetivo deste estudo foi avaliar a biocompatibilidade de nanopartícula de hidroxiapatita adicionada ao gel clareador peróxido de carbamida 16% com flúor e sem flúor e, para tanto, foram realizados testes de citotoxidade empregando MTT. Como resultados verificou-se que as nanopartículas de hidroxiapatita, quando comparadas aos géis de peróxido de carbamida a 16% com e sem flúor, foram as menos citotóxicas (p< 0.05). As diluições citotóxicas convertidas para 70% das amostras testadas também foram comparadas através do teste de Anova com tukey. Foi possível observar que as partículas de nanoHap quando adicionadas nos géis de clareamento com e sem flúor reduziu significativamento a citotoxidade. Concluímos que o novo material proposto nesta investigação apresenta melhor biocompatibilidade do que o gel sem hidroxiapatita, acompanhado da redução da citotoxidade, tais aspectos sugerem que as nanopartículas de hidroxiapatita podem ter aplicações clínicas futuras em tecidos mineralizados em procedimentos de clareamento dental, contribuindo para a redução da sensibilidade dentária.
The dental bleaching is one of the most procedures performed at dental offices to improve the appearance of the smile. The procedure consists of the application of a bleching gel, based on carbamide peroxide or hydrogen over the teeth to be whitened. Tooth sensibility is the frequently reported adverse effect on tooth whitening and is the leading cause of demotivation by the patients. The mechanism that provides the sensibility after tooth whitening has not already been fully elucidated; however, it appears to be associated with the rapid diffusion of bleaching agents through the enamel and dentin that, because of their degree of cytotoxicity, may act attacking pulp cells, causing the sensibility. The aim of this study was to evaluate the biocompatibility of hydroxyapatite nanoparticles added to whitening gel of 16% carbamide peroxide with and without fluoride and, therefore, cytotoxicity tests were performed using MTT. As a result, it was found that the hydroxyapatite nanoparticles, compared to the gels of 16 % carbamide peroxide with and without fluoride, were less cytotoxic (p <0.05). Cytotoxic dilutions converted to 70 % of the tested samples were compared using ANOVA test with Tukey. We concluded that the new material proposed in this research has a better biocompatibility in comparison with the gel without hydroxyapatite, followed by a reduction of cytotoxicity. These aspects suggest that the hydroxyapatite nanoparticles may have clinical future applications in mineralized tissues when dental bleaching procedures are performed, contributing to the reduction of tooth sensitivity. We conclude that the new material proposed in this research has a better biocompatibility of the gel without hydroxyapatite, accompanied by a reduction of cytotoxicity, these aspects suggest that the hydroxyapatite nanoparticles may have future clinical applications in mineralized tissues in dental bleaching procedures, contributing to a reduction tooth sensitivity.
Assuntos
Animais , Camundongos , Peróxidos/toxicidade , Clareamento Dental/efeitos adversos , Ureia/toxicidade , Durapatita/toxicidade , Sensibilidade da Dentina/prevenção & controle , Nanopartículas/toxicidade , Técnicas In Vitro , Teste de Materiais , Técnicas de Cultura de Células , Dessensibilizantes Dentinários/toxicidade , Clareadores Dentários/toxicidade , Clareadores Dentários/farmacologia , Antioxidantes/toxicidadeRESUMO
Aloe vera has been used in traditional medicine for the therapy of a variety of disorders, such as wounds and burns. However, few studies have examined the antioxidant capacities of A. vera plants during different growth periods. In order to investigate the effects of growth on antioxidant activity, A. vera was prepared from 2-, 4-, 6-, 8-, and 12-month-old aloe. The extracts from 6-month-old A. vera showed the highest contents of flavonoids (9.750 mg catechin equivalent/g extract) and polyphenols (23.375 mg gallic acid equivalent/g extract) and the highest ferric reducing antioxidant power (0.047 mM ferrous sulfate equivalent/mg extract). The extract from 6-month-old A. vera exhibited the highest free radical scavenging potential, and the lowest IC50 values were found for 2,2-diphenyl-1-picrylhydrazyl (0.26 mg/ml) and alkyl radicals (0.50 mg/ml). In addition, the extract from 6-month-old A. vera showed the greatest effects on cell viability in normal liver cells. Based on these findings, the extract from 6-month-old A. vera was examined further in order to determine its protective potential against tert-butyl hydroperoxide (t-BHP)-induced oxidative stress. The extract from 6-monthold A. vera at a concentration of 0.25 mg/ml showed the highest protective activity against t-BHP-induced reactive oxygen species production. These findings suggested that harvesting regimens were critical in the regulation of effects of the bioactive potential of A. vera on antioxidant activity.
Assuntos
Aloe/química , Antioxidantes/farmacologia , Sequestradores de Radicais Livres/farmacologia , Hepatócitos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Peróxidos/toxicidade , Extratos Vegetais/farmacologia , Antioxidantes/isolamento & purificação , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Flavonoides/análise , Sequestradores de Radicais Livres/isolamento & purificação , Radicais Livres/análise , Hepatócitos/fisiologia , Humanos , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Polifenóis/análiseRESUMO
There is an urgent need for new in vitro strategies to identify neurotoxic agents with speed, reliability and respect for animal welfare. Cell models should include distinct brain cell types and represent brain microenvironment to attain higher relevance. The main goal of this study was to develop and validate a human 3D neural model containing both neurons and glial cells, applicable for toxicity testing in high-throughput platforms. To achieve this, a scalable bioprocess for neural differentiation of human NTera2/cl.D1 cells in stirred culture systems was developed. Endpoints based on neuronal- and astrocytic-specific gene expression and functionality in 3D were implemented in multi-well format and used for toxicity assessment. The prototypical neurotoxicant acrylamide affected primarily neurons, impairing synaptic function; our results suggest that gene expression of the presynaptic marker synaptophysin can be used as sensitive endpoint. Chloramphenicol, described as neurotoxicant affected both cell types, with cytoskeleton markers' expression significantly reduced, particularly in astrocytes. In conclusion, a scalable and reproducible process for production of differentiated neurospheres enriched in mature neurons and functional astrocytes was obtained. This 3D approach allowed efficient production of large numbers of human differentiated neurospheres, which in combination with gene expression and functional endpoints are a powerful cell model to evaluate human neuronal and astrocytic toxicity.
Assuntos
Astrócitos/citologia , Técnicas de Cultura de Células/métodos , Neurônios/citologia , Esferoides Celulares/citologia , Testes de Toxicidade/métodos , Acrilamida/toxicidade , Astrócitos/efeitos dos fármacos , Diferenciação Celular , Linhagem Celular Tumoral , Cloranfenicol/toxicidade , Expressão Gênica/efeitos dos fármacos , Humanos , Modelos Biológicos , Neurônios/efeitos dos fármacos , Peróxidos/toxicidade , Esferoides Celulares/efeitos dos fármacosRESUMO
OBJECTIVE: This single-blind controlled study evaluated the genotoxicity and efficacy of at-home bleaching in smokers and nonsmokers. METHODS: We selected 60 patients with central incisors A2 or darker: 30 smokers (experimental group) and 30 nonsmokers (control group). The bleaching was carried out with 10% carbamide peroxide for three hours a day for three weeks. The color was evaluated using a shade guide, Vita Bleachedguide 3D-Master, at baseline, during bleaching (first, second, and third weeks), and one week and one month after bleaching. Smears were obtained with a moistened wooden spatula from marginal gingiva. All the cytologic smears were stained with Giemsa solution. From each slide, 1000 cells were examined under 40× magnification and where micronuclei (MN) were located, they were examined under 100× magnification. The change in shade guide units at the different assessment periods and the frequency of MN were subjected to a two-way repeated measures analysis of variance and Tukey test (α=0.05). RESULTS: In both groups we detected a whitening of approximately 4 to 5 shade guide units, without color rebound after one month (p>0.05). The frequency of MN was significantly higher in the experimental group than in the control group, regardless of the bleaching treatment (p>0.001). CONCLUSION: The efficacy of bleaching does not appear to be affected by the smoking habit. Additionally, at-home bleaching did not induce DNA damage to the gingival tissue during the bleaching period.
Assuntos
Peróxidos/uso terapêutico , Fumar/efeitos adversos , Clareadores Dentários/uso terapêutico , Clareamento Dental/métodos , Ureia/análogos & derivados , Adolescente , Adulto , Peróxido de Carbamida , Dano ao DNA/efeitos dos fármacos , Feminino , Gengiva/efeitos dos fármacos , Humanos , Masculino , Testes de Mutagenicidade , Peróxidos/administração & dosagem , Peróxidos/toxicidade , Autocuidado/efeitos adversos , Autocuidado/métodos , Método Simples-Cego , Clareamento Dental/efeitos adversos , Clareadores Dentários/administração & dosagem , Clareadores Dentários/toxicidade , Descoloração de Dente/tratamento farmacológico , Resultado do Tratamento , Ureia/administração & dosagem , Ureia/uso terapêutico , Ureia/toxicidade , Adulto JovemRESUMO
Alkyl hydroperoxide reductase subunit C gene (ahpC) functions were characterized in Vibrio parahaemolyticus, a commonly occurring marine food-borne enteropathogenic bacterium. Two ahpC genes, ahpC1 (VPA1683) and ahpC2 (VP0580), encoded putative two-cysteine peroxiredoxins, which are highly similar to the homologous proteins of Vibrio vulnificus. The responses of deletion mutants of ahpC genes to various peroxides were compared with and without gene complementation and at different incubation temperatures. The growth of the ahpC1 mutant and ahpC1 ahpC2 double mutant in liquid medium was significantly inhibited by organic peroxides, cumene hydroperoxide and tert-butyl hydroperoxide. However, inhibition was higher at 12°C and 22°C than at 37°C. Inhibiting effects were prevented by the complementary ahpC1 gene. Inconsistent detoxification of H2O2 by ahpC genes was demonstrated in an agar medium but not in a liquid medium. Complementation with an ahpC2 gene partially restored the peroxidase effect in the double ahpC1 ahpC2 mutant at 22°C. This investigation reveals that ahpC1 is the chief peroxidase gene that acts against organic peroxides in V. parahaemolyticus and that the function of the ahpC genes is influenced by incubation temperature.
Assuntos
Peróxidos/metabolismo , Peroxirredoxinas/metabolismo , Subunidades Proteicas/metabolismo , Vibrio parahaemolyticus/enzimologia , Meios de Cultura/química , Deleção de Genes , Teste de Complementação Genética , Peróxidos/toxicidade , Peroxirredoxinas/genética , Subunidades Proteicas/genética , Temperatura , Vibrio parahaemolyticus/efeitos dos fármacos , Vibrio parahaemolyticus/genética , Vibrio parahaemolyticus/crescimento & desenvolvimentoRESUMO
BACKGROUND: Bronchopulmonary dysplasia is one of the main complications associated with extreme prematurity. Oxidative stress is suspected to be a trigger event of this lung disease, which is characterized by impaired alveolar development. Peroxides, mainly ascorbylperoxide and H2O2, are known contaminant of parenteral nutrition. We hypothesize that these oxidant molecules induce bronchopulmonary dysplasia development. The aim was to determine if the infusion of ascorbylperoxide, whether in presence or absence of H2O2, is associated with oxidative stress, apoptosis and loss of alveoli in the lungs of newborn guinea pigs. METHOD: Three-day-old guinea pigs received parenteral solutions containing 0, 20, 60 or 180 µM ascorbylperoxide in the presence or not of 350 µM H2O2 (concentrations similar to those measured in parenteral nutrition). After 4 days, the lungs were collected for determination of glutathione's redox potential, caspase-3 activation (an apoptosis marker), alveolarization index (by histology), activation of Nrf2 and NF?B (biological markers of oxidative stress), and IL-6 and PGJ2 levels (markers of NF?B activation). Groups were compared by ANOVA, p < 0.05. RESULTS: Loss of alveoli was associated with ascorbylperoxide in a dose-dependent manner, without an influence of H2O2. The dose-dependent activation of caspase-3 by ascorbylperoxide was lower in the presence of H2O2. Ascorbylperoxide induced an increase of redox potential in a dose-dependent manner, which reached a plateau in presence of H2O2. Nrf2 and NF?B were activated by H2O2 but not by ascorbylperoxide. CONCLUSION: Results suggest that ascorbylperoxide, generated in parenteral nutrition, is involved in the development of bronchopulmonary dysplasia, independently of the increase of the redox potential. This study underlines the importance of developing a safer formulation of parenteral nutrition.
Assuntos
Ácido Ascórbico/análogos & derivados , Glutationa/metabolismo , Pulmão/efeitos dos fármacos , Peróxidos/toxicidade , Alvéolos Pulmonares/fisiopatologia , Animais , Animais Recém-Nascidos , Ácido Ascórbico/toxicidade , Caspase 3/metabolismo , Cobaias , Peróxido de Hidrogênio/toxicidade , Interleucina-6/metabolismo , Pulmão/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , NF-kappa B/metabolismo , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Nutrição Parenteral , Prostaglandina D2/análogos & derivados , Prostaglandina D2/análiseRESUMO
Candida albicans is a well-adapted human commensal but is also a facultative pathogen that can cause superficial and systemic infections. Its remarkable capacity to thrive within the human host relies on its ability to adapt and respond to the local environment of different niches. C. albicans is able to cope with oxidative stress in a coordinated fashion via upregulation of different protective mechanisms. Here, we unravel the role of a family of glutathione peroxidase (GPx), designated Gpx31, Gpx32, and Gpx33, in oxidative stress resistance. We show that GPx activity in C. albicans is induced upon exposure to peroxides and that this enzymatic activity is required for full resistance to oxidative stress. The GPx activity relies on the presence of GPX31, with no apparent contribution from GPX32 and GPX33 during in vitro short-term (3 h) exposure to peroxides. However, a triple gpx31-33Δ/Δ mutant exhibited a more pronounced sensitivity than a single gpx31Δ/Δ mutant on solid media in the presence of oxidants, suggesting that GPX32 and GPX33 may be involved in long-term adaptation to oxidative stress. Interestingly, reintegration of a single allele of GPX31 was sufficient to restore the wild-type phenotype in both the single and triple mutants. We found that mutants lacking GPX31-33 were more susceptible to killing by phagocytic cells, suggesting that GPxs are required for full resistance to innate immune effector cells. Despite the sensitivity to oxidative stress and phagocytes, these mutants were not affected in their virulence in the chicken embryo model of candidiasis.
Assuntos
Candida albicans/efeitos dos fármacos , Candida albicans/fisiologia , Tolerância a Medicamentos , Glutationa Peroxidase/metabolismo , Oxidantes/toxicidade , Estresse Oxidativo , Peróxidos/toxicidade , Animais , Candida albicans/enzimologia , Candida albicans/genética , Células Cultivadas , Embrião de Galinha , Deleção de Genes , Glutationa Peroxidase/genética , Humanos , Macrófagos/imunologia , Macrófagos/microbiologia , Viabilidade Microbiana/efeitos dos fármacos , VirulênciaRESUMO
Resistance to therapy-mediated apoptosis in inflammatory breast cancer, an aggressive and distinct subtype of breast cancer, was recently attributed to increased superoxide dismutase (SOD) expression, glutathione (GSH) content, and decreased accumulation of reactive species. In this study, we demonstrate the unique ability of two Mn(III) N-substituted pyridylporphyrin (MnP)-based SOD mimics (MnTE-2-PyP(5+) and MnTnBuOE-2-PyP(5+)) to catalyze oxidation of ascorbate, leading to the production of excessive levels of peroxide, and in turn cell death. The accumulation of peroxide, as a consequence of MnP+ascorbate treatment, was fully reversed by the administration of exogenous catalase, showing that hydrogen peroxide is essential for cell death. Cell death as a consequence of the action of MnP+ascorbate corresponded to decreases in GSH levels, prosurvival signaling (p-NF-κB, p-ERK1/2), and in expression of X-linked inhibitor of apoptosis protein, the most potent caspase inhibitor. Although markers of classical apoptosis were observed, including PARP cleavage and annexin V staining, administration of a pan-caspase inhibitor, Q-VD-OPh, did not reverse the observed cytotoxicity. MnP+ascorbate-treated cells showed nuclear translocation of apoptosis-inducing factor, suggesting the possibility of a mechanism of caspase-independent cell death. Pharmacological ascorbate has already shown promise in recently completed phase I clinical trials, in which its oxidation and subsequent peroxide formation was catalyzed by endogenous metalloproteins. The catalysis of ascorbate oxidation by an optimized metal-based catalyst (such as MnP) carries a large therapeutic potential as an anticancer agent by itself or in combination with other modalities such as radio- and chemotherapy.
Assuntos
Apoptose/efeitos dos fármacos , Neoplasias da Mama/metabolismo , Metaloporfirinas/administração & dosagem , Espécies Reativas de Oxigênio/metabolismo , Ácido Ascórbico/metabolismo , Neoplasias da Mama/patologia , Caspases/genética , Feminino , Humanos , Peróxido de Hidrogênio/metabolismo , NF-kappa B/metabolismo , Oxirredução , Peróxidos/metabolismo , Peróxidos/toxicidade , Espécies Reativas de Oxigênio/toxicidade , Superóxido Dismutase/metabolismoRESUMO
Sugarcane molasses is a rich source of antioxidant materials with peroxyl radical scavenging effects. To explore the potent antioxidant activity of sugarcane molasses against 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced peroxyl radicals, 7 methanolic fractions of sugarcane molasses (F1-F7) were separated via bioassay-guided fractionation and evaluated by oxygen radical absorbance capacity (ORAC), cellular antioxidant activity (CAA), and oxidative DNA damage protective activity assays. The ORAC values of sugarcane molasses fractions ranged from 4399 to 6,266 µmol TE/g, whilst the EC50 values for CAA ranged from 3.7 to 5.9 µg/ml. Moreover, it was found that sugarcane molasses fractions, particularly F6 and F7, could protect against oxidative DNA damage caused by peroxyl radicals at an effective concentration of 100 µg/ml. Ten phenolic constituents were identified in the fractions, including known antioxidative compounds, viz., schaftoside, isoschaftoside, ferulic acid, p-coumaric acid, and p-hydroxybenzaldehyde.
Assuntos
Amidinas/química , Sequestradores de Radicais Livres/química , Melaço/análise , Peróxidos/química , Extratos Vegetais/química , Saccharum/química , Amidinas/toxicidade , Linhagem Celular , Dano ao DNA/efeitos dos fármacos , Sequestradores de Radicais Livres/farmacologia , Humanos , Peróxidos/toxicidade , Extratos Vegetais/farmacologiaRESUMO
The aim of this study was to evaluate the possibility of fluoride solutions applied to enamel to protect pulp cells against the trans-enamel and transdentinal cytotoxicity of a 16% carbamide peroxide (CP) bleaching gel. The CP gel was applied to enamel/dentin discs adapted to aicial pulp chambers (8 h/day) during 1, 7 or 14 days, followed by fluoride (0.05% or 0.2%) application for 1 min. The extracts (culture medium in contact with dentin) were applied to MDPC-23 cells for 1 h, and cell metabolism (MTT assay), alkaline phosphatase (ALP) activity and cell membrane damage (flow cytometry) were analyzed. Knoop microhardness of enamel was also evaluated. Data were analyzed statistically by ANOVA and Kruskal-Wallis tests (α=0.05). For the MTT assay and ALP activity, significant reductions between the control and the bleached groups were observed (p<0.05). No statistically significant difference occurred among bleached groups (p>0.05), regardless of fluoride application or treatment days. Flow cytometry analysis demonstrated 30% of cell membrane damage in all bleached groups. After 14 days of treatment, the fluoride-treated enamel presented significantly higher microhardness values than the bleached-only group (p<0.05). It was concluded that, regardless of the increase in enamel hardness due to the application of fluoride solutions, the treated enamel surface did not prevent the toxic effects caused by the 16% CP gel to odontoblast-like cells.
Resumo O objetivo do presente estudo foi avaliar o possível efeito protetor de soluções fluoretadas aplicadas sobre o esmalte dentário frente à citotoxicidade trans-amelodentinária de um gel clareador com 16% de peróxido de carbamida (PC). O gel de PC foi aplicado sobre discos de esmalte/dentina adaptados a câmaras pulpares aiciais (8 h/dia) durante períodos de 1, 7 ou 14 dias, seguido de aplicação de soluções fluoretadas (0,05% ou 0,2%) durante 1 min. Os extratos (meio de cultura em contato com a dentina) foram aplicados sobre células MDPC-23 durante 1 h, seguido de análise do metabolismo celular (teste do MTT), atividade de fosfatase alcalina (ALP) e danos à membrana celular (citometria de fluxo). A microdureza Knoop do esmalte dental foi avaliada. Os dados foram analisados pelos testes de ANOVA e Kruskal-Wallis. Para o teste do MTT e atividade de ALP, redução significante entre os grupos controle e clareados foram observados (p<0,05). Nenhuma diferença entre os grupos clareados foi observada (p>0,05), independente da aplicação das soluções fluoretadas ou tempo de tratamento. A análise por citometria de fluxo demonstrou lesão à membrana celular em torno de 30% para todos os grupos clareados. Após 14 dias de tratamento, os espécimes clareados e fluoretados apresentaram aumento significante na microdureza do esmalte (p<0,05). Pôde-se concluir que apesar do aumento na dureza do esmalte decorrente da aplicação das soluções fluoretadas, este tratamento não preveniu os efeitos tóxicos causados pelo gel com 16% de PC sobre as células odontoblastóides. .
Assuntos
Animais , Bovinos , Esmalte Dentário/efeitos dos fármacos , Polpa Dentária/efeitos dos fármacos , Fluoretos/farmacologia , Peróxidos/toxicidade , Substâncias Protetoras/farmacologia , Clareadores Dentários/toxicidade , Ureia/análogos & derivados , Fosfatase Alcalina/efeitos dos fármacos , Linhagem Celular , Membrana Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Corantes , Cavidade Pulpar/efeitos dos fármacos , Polpa Dentária/citologia , Dentina/efeitos dos fármacos , Dureza , Odontoblastos/efeitos dos fármacos , Odontoblastos/metabolismo , Propídio , Succinato Desidrogenase/efeitos dos fármacos , Fatores de Tempo , Ureia/toxicidadeRESUMO
Oxidant imbalance is one of the causative mechanisms of aluminum-induced neurotoxicity. In this study, we investigated aluminum-induced oxidant imbalance in non-neuronal tissues (liver, kidney and testis) and temporal cortex in rats. The differences in adaptations to superoxide and peroxide handling capacities (SPHC) of studied organs due to aluminum insult were also evaluated. Male Wistar rats were exposed to aluminum (10 mg/Kg body wt/day) for 4 weeks through orogastric intubation. Liver showed significant decrease in reduced glutathione level, while significant alteration in lipid peroxidation was observed in temporal cortex in aluminium-exposed animals. Superoxide dismutase activity was significantly altered in liver and temporal cortex and catalase activity significantly reduced in the liver due to aluminum exposure, while glutathione reductase and glutathione peroxidase activities were altered in all the tested organs. Among the organs, glutathione-independent SPHC was relatively higher in liver and kidney, while glutathione-dependent SPHC was relatively higher in testis and temporal cortex. As compared to control, aluminum-exposed rats demonstrated reduction in glutathione-dependent SPHC in temporal cortex and increment of the same in testis, while increment in glutathione-independent SPHC was observed in liver. In conclusion, aluminum-induced alteration in oxidant handling capacity could be the cause of oxidative stress both in the neuronal and non-neuronal tissues.