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1.
Front Microbiol ; 12: 619313, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33746916

RESUMO

High-throughput screening methodologies to estimate lipid content in oleaginous yeasts use Nile red fluorescence in a given solvent and optimized excitation/emission wavelengths. However, Nile red fluorescence stabilization has been poorly analyzed, and high variability occurs when relative fluorescence is measured immediately or a few minutes after dye addition. The aim of this work was to analyze the fluorescence of Nile red at different incubation times using a variety of solvents and oleaginous/non-oleaginous yeast strains. We showed that fluorescence stabilization occurs between 20 and 30 min, depending on the strain and solvent. Therefore, we suggest that fluorescence measurements should be followed until stabilization, where Relative Fluorescence Units should be considered after stabilization for lipid content estimation.

2.
Oxid Med Cell Longev ; 2019: 2510936, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31772702

RESUMO

Tellurium is a rare element that has been regarded as a toxic, nonessential element, and its biological role is not clearly established. In addition, the biological effects of elemental tellurium and some of its organic and inorganic derivatives have been studied, leading to a set of interesting and promising applications. Diphenyl ditelluride (DPDT), an organic tellurium derivate, showed antioxidant, antigenotoxic, antimutagenic, and anticancer properties. The antioxidant and prooxidant properties of DPDT are complex and depend on experimental conditions, which may explain the contradictory reports of these properties. In addition, DPDT may exert its effects through different pathways, including distinct ones to those responsible for chemotherapy resistance phenotypes: transcription factors, membrane receptors, adhesion, structural molecules, cell cycle regulatory components, and apoptosis pathways. This review aims to present recent advances in our understanding of the biological effects, therapeutic potential, and safety of DPDT treatment. Moreover, original results demonstrating the cytotoxic effects of DPDT in different mammalian cell lines and systems biology analysis are included, and emerging approaches for possible future applications are inferred.


Assuntos
Antioxidantes/uso terapêutico , Derivados de Benzeno/uso terapêutico , Compostos Organometálicos/uso terapêutico , Telúrio/química , Antioxidantes/farmacologia , Derivados de Benzeno/farmacologia , Humanos , Compostos Organometálicos/farmacologia , Oxirredução
3.
Antiviral Res ; 172: 104615, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31580916

RESUMO

Epstein-Barr virus (EBV) is a widely distributed gamma-herpesvirus that has been associated with various cancers mainly from lymphocytic and epithelial origin. Although EBV-mediated oncogenesis has been associated with viral oncogenes expressed during latency, a growing set of evidence suggested that antiviral treatments directed against EBV lytic phase may contribute to prevent some forms of cancers, including EBV-positive Post-Transplant Lymphoproliferative Diseases. It is shown here that dipyridamole (DIP), a safe drug with favorable and broad pharmacological properties, inhibits EBV reactivation from B-cell lines. DIP repressed immediate early and early genes expression mostly through its ability to inhibit nucleoside uptake. Considering its wide clinical use, DIP repurposing could shortly be evaluated, alone or in combination with other antivirals, to treat EBV-related diseases where lytic replication plays a deleterious role.


Assuntos
Dipiridamol/farmacologia , Herpesvirus Humano 4/efeitos dos fármacos , Ativação Viral/efeitos dos fármacos , Antivirais/farmacologia , Linfócitos B/metabolismo , Linfócitos B/virologia , Linhagem Celular , DNA Viral/efeitos dos fármacos , Reposicionamento de Medicamentos , Infecções por Vírus Epstein-Barr/tratamento farmacológico , Expressão Gênica/efeitos dos fármacos , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/metabolismo , Humanos , Nucleosídeos/metabolismo , Latência Viral/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos
4.
Mutat Res Genet Toxicol Environ Mutagen ; 836(Pt B): 24-35, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30442341

RESUMO

DNA and chromosomal damage in individuals occupationally exposed to coal mining residues have repeatedly been reported in lymphocytes and epithelial cells, suggesting a systemic exposure-response in which generation of oxidative damage may play a major role. Nevertheless, the understanding of this mechanism is still incomplete, particularly in regard to environmental exposures. This study aimed to evaluate DNA damage using the cytome assay (BMN-cyt) in buccal cells and its relation to primary and oxidative DNA damage in lymphocytes, assessed by the high-throughput alkaline and modified (FPG-ENDO III) Comet assay in individuals with environmental exposure to coal mining residues in northern Colombia. Considering metals from coal mining activities as the main source of reactive oxygen species (ROS) generation, the concentrations of inorganic elements in blood samples was also assessed. The analysis revealed that frequencies of BMN-cyt parameters related to DNA damage (micronuclei), cytokinesis (binucleated cells) and cell death (condensed chromatin, karyorrhexis, pyknosis and karyolysis) were significantly higher in individuals that were environmentally exposed to coal compared to the unexposed group. The level of % Tail DNA in the alkaline and the modified Comet assay was 4.0 and 4.3 times higher among exposed individuals than in unexposed controls respectively. Increased MN frequencies in buccal cells were correlated with increased %Tail DNA in alkaline and FPG Comet assay. Additionally, exposed individuals had higher concentrations of Cr, Ni, Mn, and Br in the blood compared to unexposed controls. %Tail DNA in alkaline Comet assay was highly correlated with Al, Mn, and Br concentrations, while %Tail DNA in the FPG Comet assay correlated with Mn levels. These results suggest that oxidative damage, particularly purine oxidation, may play an essential role in DNA damage in individuals exposed to coal residues and that some inorganic elements are related to this effect.


Assuntos
Minas de Carvão , Ensaio Cometa/métodos , Dano ao DNA , Monitoramento Ambiental/métodos , Testes para Micronúcleos/métodos , Mucosa Bucal/patologia , Exposição Ocupacional/análise , Células Cultivadas , Humanos , Linfócitos/efeitos dos fármacos , Linfócitos/metabolismo , Linfócitos/patologia , Mucosa Bucal/efeitos dos fármacos , Mucosa Bucal/metabolismo , Exposição Ocupacional/efeitos adversos
5.
Chemosphere ; 206: 203-216, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29751246

RESUMO

During coal surface mining, several activities such as drilling, blasting, loading, and transport produce large quantities of particulate matter (PM) that is directly emitted into the atmosphere. Occupational exposure to this PM has been associated with an increase of DNA damage, but there is a scarcity of data examining the impact of these industrial operations in cytogenetic endpoints frequency and cancer risk of potentially exposed surrounding populations. In this study, we used a Geographic Information Systems (GIS) approach and Inverse Distance Weighting (IDW) methods to perform a spatial and statistical analysis to explore whether exposure to PM2.5 and PM10 pollution, and additional factors, including the enrichment of the PM with inorganic elements, contribute to cytogenetic damage in residents living in proximity to an open-pit coal mining area. Results showed a spatial relationship between exposure to elevated concentrations of PM2.5, PM10 and micronuclei frequency in binucleated (MNBN) and mononucleated (MNMONO) cells. Active pits, disposal, and storage areas could be identified as the possible emission sources of combustion elements. Mining activities were also correlated with increased concentrations of highly enriched elements like S, Cu and Cr in the atmosphere, corroborating its role in the inorganic elements pollution around coal mines. Elements enriched in the PM2.5 fraction contributed to increasing of MNBN but seems to be more related to increased MNMONO frequencies and DNA damage accumulated in vivo. The combined use of GIS and IDW methods could represent an important tool for monitoring potential cancer risk associated to dynamically distributed variables like the PM.


Assuntos
Poluição do Ar/análise , Minas de Carvão/normas , Ciências da Terra/métodos , Monitoramento Ambiental/métodos , Material Particulado/química , Humanos
6.
Food Chem Toxicol ; 112: 383-392, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29337231

RESUMO

Pleurotus sajor-caju (PSC) is an edible mushroom used in food supplements, presenting antitumor properties through induction of cell death pathways. The PSC potential against colorectal cancer was analyzed by exposing HCT116wt cells to different PSC extracts. The PSC n-hexane extract (PSC-hex) showed the highest cytotoxicity effect (IC50 value 0.05 mg/mL). The observed cytotoxicity was then associated to apoptosis-promoting and cell cycle-arrest pathways. PSC-hex was able to induce apoptosis related to breakdown of mitochondrial membrane potential and ROS generation. The absence of cytotoxicity in HTC116-p53 and HTC116-Bax cells, alongside with an increase in p53, Bax and Caspase-3 expression, and decrease in Bcl-2 expression, supports that the pro-apoptotic effect is probably induced through a p53 associated pathway. PSC-hex induced cell cycle arrest at G2/M in HCT116wt without cytotoxicity in HTC116-p21 cells. These findings suggest that a p21/p53 cell cycle regulation pathway is probably disrupted by compounds present on PSC-hex. Identification of the major components was then performed with ergosta-5,7,22-trien-3ß-ol representing 30.6% of total weight. In silico docking studies of ergosta-5,7,22-trien-3ß against Bcl-2 were performed and results show a credible interaction with the Bcl-2 hydrophobic cleft. The results show that PSC-hex can be used as supplementary food for adjuvant therapy in colorectal carcinoma.


Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias Colorretais/terapia , Suplementos Nutricionais , Pleurotus/química , Antineoplásicos/isolamento & purificação , Caspase 3/metabolismo , Divisão Celular/efeitos dos fármacos , Linhagem Celular , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Ergosterol/análogos & derivados , Ergosterol/isolamento & purificação , Ergosterol/farmacologia , Fase G2/efeitos dos fármacos , Células HCT116 , Humanos , Interações Hidrofóbicas e Hidrofílicas , Concentração Inibidora 50 , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Simulação de Acoplamento Molecular , Proteínas Proto-Oncogênicas c-bcl-2/antagonistas & inibidores , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteína Supressora de Tumor p53/metabolismo
7.
Food Chem ; 245: 150-158, 2018 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-29287356

RESUMO

A study with Pleurotus sajor-caju was conducted to: evaluate the nutritional and chemical composition of the fruiting bodies; optimize the preparation of bioactive phenolic extracts; and characterize the optimized extract in terms of bioactive compounds and properties. P. sajor-caju revealed an equilibrated nutritional composition with the presence of hydrophilic (sugars and organic acids) and lipophilic (tocopherols and PUFA) compounds. p-Hydroxybenzoic, p-coumaric and cinnamic acids were identified in the extract obtained with ethanol (30g/l ratio) at 55°C for 85min. This extract showed antioxidant properties (mainly reducing power and lipid peroxidation inhibition), antibacterial activity against MRSA and MSSA and cytotoxicity against NCI-H460, MCF-7 and HeLa. Furthermore, as the extract showed capacity to inhibit NO production in Raw 264.7 macrophages, molecular docking studies were performed to provide insights into the anti-inflammatory mechanism of action, through COX-2 inhibition by the phenolic acids identified.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Antioxidantes/farmacologia , Valor Nutritivo , Pleurotus/química , Anti-Inflamatórios não Esteroides/química , Antineoplásicos/química , Antineoplásicos/farmacologia , Antioxidantes/química , Etanol/química , Células HeLa , Humanos , Células MCF-7 , Simulação de Acoplamento Molecular , Tocoferóis/análise
8.
Ecotoxicol Environ Saf ; 148: 453-466, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29102906

RESUMO

Epidemiological studies indicate that living in proximity to coal mines is correlated with numerous diseases including cancer, and that exposure to PM10 and PM2.5 components could be associated with this phenomenon. However, the understanding of the mechanisms by which PM exerts its adverse effects is still incomplete and comes mainly from studies in occupationally exposed populations. The aims of this study were to: (1) evaluate DNA damage in lymphocytes assessing the cytokinesis-block micronucleus cytome assay (CBMN-cyt) parameters; (2) identify aneugenic or clastogenic effects in lymphocytes of exposed populations using CREST immunostaining for micronuclei; (3) evaluate multi-elemental composition of atmospheric particulate matter; and (4) verify relation between the DNA damage and PM2.5 and PM10 levels around the mining area. Analysis revealed a significant increase in micronuclei frequency in binucleated (MNBN) and mononucleated (MNMONO) cells of individuals with residential proximity to open-pit coal mines compared to residents from non-mining areas. Correlation analysis demonstrated a highly significant association between PM2.5 levels, MNBN frequencies and CREST+ micronuclei induction in exposed residents. These results suggest that PM2.5 fraction generated in coal mining activities may induce whole chromosome loss (aneuploidy) preferentially, although there are also chromosome breaks. Analysis of the chemical composition of PM2.5 by PIXE demonstrated that Si, S, K and Cr concentrations varied significantly between coal mining and reference areas. Enrichment factor values (EF) showed that S, Cr and Cu were highly enriched in the coal mining areas. Compared to reference area, mining regions had also higher concentrations of extractable organic matter (EOM) related to nonpolar and polar compounds. Our results demonstrate that PM2.5 fraction represents the most important health risk for residents living near open-pit mines, underscoring the need for incorporation of ambient air standards based on PM2.5 measures in coal mining areas.


Assuntos
Poluentes Atmosféricos/toxicidade , Minas de Carvão , Dano ao DNA , Exposição Ocupacional/efeitos adversos , Material Particulado/toxicidade , Adolescente , Adulto , Núcleo Celular/efeitos dos fármacos , Carvão Mineral , Colômbia , Monitoramento Ambiental , Feminino , Humanos , Masculino , Testes para Micronúcleos , Pessoa de Meia-Idade , Adulto Jovem
9.
Front Microbiol ; 8: 1776, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29018411

RESUMO

A high throughput screening (HTS) methodology for evaluation of cellular lipid content based on Nile red fluorescence reads using black background 96-wells test plates and a plate reader equipment allowed the rapid intracellular lipid estimation of strains from a Brazilian phylloplane yeast collection. A new oleaginous yeast, Meyerozyma guilliermondii BI281A, was selected, for which the gravimetric determination of total lipids relative to dry weight was 52.38% for glucose or 34.97% for pure glycerol. The lipid production was optimized obtaining 108 mg/L of neutral lipids using pure glycerol as carbon source, and the strain proved capable of accumulating oil using raw glycerol from a biodiesel refinery. The lipid profile showed monounsaturated fatty acids (MUFA) varying between 56 or 74% in pure or raw glycerol, respectively. M. guilliermondii BI281A bears potential as a new biodiesel feedstock.

10.
An Acad Bras Cienc ; 89(1): 155-161, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28273245

RESUMO

Several studies have shown that a high consumption of vegetables and fruits is consistently associated with a low risk of oxidative stress-induced diseases, which includes some degenerative diseases such as amyotrophic lateral sclerosis, Alzheimer and Parkinson. Therefore, the objective of this study is to verify the effects of conventional and organic grape juice in the modulation of the neurotrophic factor (BDNF) and astrocytic markers protein (S100B) in hippocampus and frontal cortex of Wistar rats. In this study, 24 male Wistar rats were divided into three groups. To the first one, it was given organic purple grape juice; to the second, conventional grape juice, while the last one received only saline. After 30 days, all rats were sacrificed and hippocampus and frontal cortex were dissected. The animals that received organic and conventional grape juice showed, in frontal cortex, an elevated BNDF levels in relation to saline group. However, S100B levels did not change. These results showed that grape juices are able to modulate important marker in brain tissue, and could be an important factor to prevent brain diseases.


Assuntos
Fator Neurotrófico Derivado do Encéfalo/análise , Lobo Frontal/química , Sucos de Frutas e Vegetais , Hipocampo/química , Subunidade beta da Proteína Ligante de Cálcio S100/análise , Vitis/química , Animais , Antioxidantes/farmacologia , Fator Neurotrófico Derivado do Encéfalo/efeitos dos fármacos , Alimentos Orgânicos , Lobo Frontal/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Masculino , Distribuição Aleatória , Ratos Wistar , Valores de Referência , Reprodutibilidade dos Testes , Subunidade beta da Proteína Ligante de Cálcio S100/efeitos dos fármacos
11.
An. acad. bras. ciênc ; 89(1): 155-161, Jan,-Mar. 2017. tab, graf
Artigo em Inglês | LILACS | ID: biblio-886639

RESUMO

ABSTRACT Several studies have shown that a high consumption of vegetables and fruits is consistently associated with a low risk of oxidative stress-induced diseases, which includes some degenerative diseases such as amyotrophic lateral sclerosis, Alzheimer and Parkinson. Therefore, the objective of this study is to verify the effects of conventional and organic grape juice in the modulation of the neurotrophic factor (BDNF) and astrocytic markers protein (S100B) in hippocampus and frontal cortex of Wistar rats. In this study, 24 male Wistar rats were divided into three groups. To the first one, it was given organic purple grape juice; to the second, conventional grape juice, while the last one received only saline. After 30 days, all rats were sacrificed and hippocampus and frontal cortex were dissected. The animals that received organic and conventional grape juice showed, in frontal cortex, an elevated BNDF levels in relation to saline group. However, S100B levels did not change. These results showed that grape juices are able to modulate important marker in brain tissue, and could be an important factor to prevent brain diseases.


Assuntos
Animais , Masculino , Fator Neurotrófico Derivado do Encéfalo/análise , Vitis/química , Subunidade beta da Proteína Ligante de Cálcio S100/análise , Sucos de Frutas e Vegetais , Lobo Frontal/química , Hipocampo/química , Valores de Referência , Distribuição Aleatória , Reprodutibilidade dos Testes , Ratos Wistar , Fator Neurotrófico Derivado do Encéfalo/efeitos dos fármacos , Alimentos Orgânicos , Subunidade beta da Proteína Ligante de Cálcio S100/efeitos dos fármacos , Lobo Frontal/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Antioxidantes/farmacologia
12.
An Acad Bras Cienc ; 88(4): 2247-2255, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27991961

RESUMO

Vitis labrusca L. is the main species used for wine and juice production in Brazil. The grapevine leaves can be used both as functional foods and as cheapest sources for the extraction of phenolic compounds. Besides the antioxidant activity, grapevine leaves exhibited significant anti-inflammatory activity. Therefore, the aim of this study was to develop and validate an analytical methodology to determine the metals selenium (96Se), chromium (53Cr), nickel (62Ni), cadmium (111Cd) and lead (206Pb) in 30 samples of grapevine leaf extracts (Vitis labrusca, Bordo cultivar) using inductively coupled plasma mass spectrometry (ICP-MS). To obtain the grapevine leaf extracts the samples were milled, weighed and digested in microwave oven with nitric acid. The method showed linearity, precision, accuracy and limits of quantification and detection acceptable for INMETRO protocol validation of analytical methods. Therefore, the method using ICP-MS was developed and validated to determine metals concentrations in grapevine leaves of Vitis labrusca L. and the proposed method could be applied in routine analytical laboratory.


Assuntos
Metais/análise , Extratos Vegetais/química , Folhas de Planta/química , Vitis/química , Brasil , Cádmio/análise , Cromo/análise , Chumbo/análise , Espectrometria de Massas/métodos , Níquel/análise , Fenóis/isolamento & purificação , Selênio/análise
13.
Artigo em Inglês | MEDLINE | ID: mdl-27637484

RESUMO

Increasing evidence suggest that occupational exposure to open-cast coal mining residues like dust particles, heavy metals and Polycyclic Aromatic Hydrocarbons (PAHs) may cause a wide range of DNA damage and genomic instability that could be associated to initial steps in cancer development and other work-related diseases. The aim of our study was to evaluate if key polymorphisms in metabolism genes CYP1A1Msp1, GSTM1Null, GSTT1Null and DNA repair genes XRCC1Arg194Trp and hOGG1Ser326Cys could modify individual susceptibility to adverse coal exposure effects, considering the DNA damage (Comet assay) and micronucleus formation in lymphocytes (CBMN) and buccal mucosa cells (BMNCyt) as endpoints for genotoxicity. The study population is comprised of 200 healthy male subjects, 100 open-cast coal-mining workers from "El Cerrejón" (world's largest open-cast coal mine located in Guajira - Colombia) and 100 non-exposed referents from general population. The data revealed a significant increase of CBMN frequency in peripheral lymphocytes of occupationally exposed workers carrying the wild-type variant of GSTT1 (+) gene. Exposed subjects carrying GSTT1null polymorphism showed a lower micronucleus frequency compared with their positive counterparts (FR: 0.83; P=0.04), while BMNCyt, frequency and Comet assay parameters in lymphocytes: Damage Index (DI) and percentage of DNA in the tail (Tail % DNA) were significantly higher in exposed workers with the GSTM1Null polymorphism. Other exfoliated buccal mucosa abnormalities related to cell death (Karyorrhexis and Karyolysis) were increased in GSTT/M1Null carriers. Nuclear buds were significantly higher in workers carrying the CYP1A1Msp1 (m1/m2, m2/m2) allele. Moreover, BMNCyt frequency and Comet assay parameters were significantly lower in exposed carriers of XRCC1Arg194Trp (Arg/Trp, Trp/Trp) and hOGG1Ser326Cys (Ser/Cys, Cys/Cys), thereby providing new data to the increasing evidence about the protective role of these polymorphisms. This modulation may involve specific and differentiated pathways in different tissues that also may cause a differential sensitivity related to differential induction of some enzymes.


Assuntos
Minas de Carvão/métodos , Reparo do DNA/genética , Exposição Ocupacional/efeitos adversos , Polimorfismo de Nucleotídeo Único , Adulto , Estudos de Casos e Controles , Colômbia , Estudos Transversais , Citocromo P-450 CYP1A1/genética , Dano ao DNA , DNA Glicosilases/genética , Proteínas de Ligação a DNA/genética , Estudos de Associação Genética , Glutationa Transferase/genética , Heterozigoto , Humanos , Inativação Metabólica/genética , Masculino , Pessoa de Meia-Idade , Proteína 1 Complementadora Cruzada de Reparo de Raio-X , Adulto Jovem
14.
An Acad Bras Cienc ; 88(3 Suppl): 1655-1674, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27556332

RESUMO

Parkinson's disease is characterized by the death of dopaminergic neurons, mainly in the substantia nigra, and causes serious locomotor dysfunctions. It is likely that the oxidative damage to cellular biomolecules is among the leading causes of neurodegeneration that occurs in the disease. Selenium is an essential mineral for proper functioning of the brain, and mainly due to its antioxidant activity, it is possible to exert a special role in the prevention and in the nutritional management of Parkinson's disease. Currently, few researchers have investigated the effects of selenium on Parkinson´s disease. However, it is known that very high or very low body levels of selenium can (possibly) contribute to the pathogenesis of Parkinson's disease, because this imbalance results in increased levels of oxidative stress. Therefore, the aim of this work is to review and discuss studies that have addressed these topics and to finally associate the information obtained from them so that these data and associations serve as input to new research.


Assuntos
Estresse Oxidativo , Doença de Parkinson/etiologia , Selênio/fisiologia , Encéfalo/fisiologia , Neurônios Dopaminérgicos/patologia , Humanos , Doença de Parkinson/prevenção & controle , Substância Negra/patologia
15.
J Pharm Pharmacol ; 68(8): 1073-84, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27283471

RESUMO

OBJECTIVE: Red propolis, an exclusive variety of propolis found in the northeast of Brazil has shown to present antitumour activity, among several other biological properties. This article aimed to help to evaluate the underlying molecular mechanisms of the potential anticancer effects of red propolis on tumour, Hep-2, and non-tumour cells, Hek-293. METHODS: Differentially expressed proteins in human cell lines were identified through label-free quantitative MS-based proteomic platform, and cells were stained with Giemsa to show morphological changes. KEY FINDINGS: A total of 1336 and 773 proteins were identified for Hep-2 and Hek-293, respectively. Among the proteins here identified, 16 were regulated in the Hep-2 cell line and 04 proteins in the Hek-293 line. Over a total of 2000 proteins were identified under MS analysis, and approximately 1% presented differential expression patterns. The GO annotation using Protein Analysis THrough Evolutionary Relationships classification system revealed predominant molecular function of catalytic activity, and among the biological processes, the most prominent was associated to cell metabolism. CONCLUSION: The proteomic profile here presented should help to elucidate further molecular mechanisms involved in inhibition of cancer cell proliferation by red propolis, which remain unclear to date.


Assuntos
Antineoplásicos/farmacologia , Apiterapia , Neoplasias/metabolismo , Própole/farmacologia , Proteoma/metabolismo , Antineoplásicos/uso terapêutico , Brasil , Linhagem Celular Tumoral , Cromatografia Líquida , Células HEK293 , Humanos , Espectrometria de Massas , Neoplasias/tratamento farmacológico , Própole/uso terapêutico , Proteômica
16.
Artigo em Inglês | MEDLINE | ID: mdl-27042187

RESUMO

Garcinia mangostana, popularly known as "mangosteen fruit," originates from Southeast Asia and came to Brazil about 80 years ago where it mainly grows in the states of Pará and Bahia. Although mangosteen or its extracts have been used for ages in Asian folk medicine, data on its potential genotoxicity is missing. We, therefore, evaluated genotoxicity/mutagenicity of hydroethanolic mangosteen extract [HEGM, 10 to 640 µg/mL] in established test assays (Comet assay, micronucleus test, and Salmonella/microsome test). In the Comet assay, HEGM-exposed human leukocytes showed no DNA damage. No significant HEGM-induced mutation in TA98 and TA100 strains of Salmonella typhimurium (with or without metabolic activation) was observed and HEGM-exposed human lymphocytes had no increase of micronuclei. However, HEGM suggested exposure concentration-dependent antigenotoxic potential in leukocytes and antioxidant potential in the yeast Saccharomyces cerevisiae. HEGM preloading effectively protected against H2O2-induced DNA damage in leukocytes (Comet assay). Preloading of yeast with HEGM for up to 4 h significantly protected the cells from lethality of chronic H2O2-exposure, as expressed in better survival. Absence of genotoxicity and demonstration of an antigenotoxic and antioxidant potential suggest that HEGM or some substances contained in it may hold promise for pharmaceutical or nutraceutical application.

17.
Oncotarget ; 7(18): 25885-901, 2016 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-27029031

RESUMO

Trabectedin (Yondelis®, ecteinascidin-743, ET-743) is a marine-derived natural product approved for treatment of advanced soft tissue sarcoma and relapsed platinum-sensitive ovarian cancer. Lurbinectedin is a novel anticancer agent structurally related to trabectedin. Both ecteinascidins generate DNA double-strand breaks that are processed through homologous recombination repair (HRR), thereby rendering HRR-deficient cells particularly sensitive. We here characterize the DNA damage response (DDR) to trabectedin and lurbinectedin in HeLa cells. Our results show that both compounds activate the ATM/Chk2 (ataxia-telangiectasia mutated/checkpoint kinase 2) and ATR/Chk1 (ATM and RAD3-related/checkpoint kinase 1) pathways. Interestingly, pharmacological inhibition of Chk1/2, ATR or ATM is not accompanied by any significant improvement of the cytotoxic activity of the ecteinascidins while dual inhibition of ATM and ATR strongly potentiates it. Accordingly, concomitant inhibition of both ATR and ATM is an absolute requirement to efficiently block the formation of γ-H2AX, MDC1, BRCA1 and Rad51 foci following exposure to the ecteinascidins. These results are not restricted to HeLa cells, but are shared by cisplatin-sensitive and -resistant ovarian carcinoma cells. Together, our data identify ATR and ATM as central coordinators of the DDR to ecteinascidins and provide a mechanistic rationale for combining these compounds with ATR and ATM inhibitors.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Proteínas Mutadas de Ataxia Telangiectasia/antagonistas & inibidores , Carbolinas/farmacologia , Dioxóis/farmacologia , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Tetra-Hidroisoquinolinas/farmacologia , Antineoplásicos Alquilantes/farmacologia , Dano ao DNA/efeitos dos fármacos , Células HeLa , Humanos , Reparo de DNA por Recombinação/efeitos dos fármacos , Trabectedina
18.
An Acad Bras Cienc ; 88(1): 281-92, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26839997

RESUMO

Salvia officinalis (Lamiaceae) has been used in south of Brazil as a diary homemade, in food condiment and tea-beverage used for the treatment of several disorders. The objective of this study was to characterize chemical compounds in the hydroalcoholic (ExtHS) and aqueous (ExtAS) extract from Salvia officinalis (L.) by gas chromatography-mass spectrometry (GC-MS) and by high-resolution electrospray ionization mass spectrometry (ESI-QTOF MS/MS), evaluate in vitro ability to scavenge the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), catalase (CAT-like) and superoxide dismutase (SOD-like) activity, moreover cytotoxic by MTT assay, alterations on cell morphology by giemsa and apoptotic-induced mechanism for annexin V/propidium iodide. Chemical identification sage extracts revealed the presence of acids and phenolic compounds. In vitro antioxidant analysis for both extracts indicated promising activities. The cytotoxic assays using tumor (Hep-2, HeLa, A-549, HT-29 and A-375) and in non-tumor (HEK-293 and MRC-5), showed selectivity for tumor cell lines. Immunocytochemistry presenting a majority of tumor cells at late stages of the apoptotic process and necrosis. Given the results presented here, Brazilian Salvia officinalis (L.) used as condiment and tea, may protect the body against some disease, in particularly those where oxidative stress is involved, like neurodegenerative disorders, inflammation and cancer.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Antioxidantes/farmacologia , Extratos Vegetais/farmacologia , Salvia officinalis/química , Linhagem Celular Tumoral/efeitos dos fármacos , Proliferação de Células , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Imuno-Histoquímica , Estresse Oxidativo/efeitos dos fármacos
19.
An Acad Bras Cienc ; 88(1): 349-60, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26839999

RESUMO

Agricultural workers involved in the harvest of tobacco crops are regularly exposed to large quantities of pesticides. In order to determine how this exposure to pesticides induces genetic alterations in these workers, blood samples were obtained from 77 exposed individuals, as well as from 60 unexposed subjects. DNA damage was analyzed by the Comet assay and by the micronucleus (MN) test. The antioxidant profile was evaluated by activity of superoxide dismutase (SOD), and the polymorphism of gene PON1 was used as a susceptibility biomarker. The content of inorganic elements in the blood samples was determined by PIXE analysis. Our results demonstrated that the damage frequency, damage index, the MN frequency, and the SOD activity were significantly elevated in the exposed relative to the unexposed group. A modulation of the MN results for the PON1 gene was observed in the exposed group. The concentrations of inorganic elements in the exposed group were higher compared to those of the unexposed group. In this study, we observed that genetic damage, and change in oxidative balance were induced by the exposure of workers to complex mixtures of pesticides in the presence of inorganic compounds, whereby an influence of the genotype was evident.


Assuntos
Dano ao DNA/efeitos dos fármacos , Fazendeiros/estatística & dados numéricos , Exposição Ocupacional/análise , Praguicidas/toxicidade , Superóxido Dismutase/sangue , Adulto , Biomarcadores/sangue , Ensaio Cometa , Dano ao DNA/genética , Diagnóstico Precoce , Feminino , Predisposição Genética para Doença/genética , Genótipo , Humanos , Masculino , Testes para Micronúcleos , Pessoa de Meia-Idade , Polimorfismo Genético , Nicotiana
20.
Int J Radiat Biol ; 91(11): 891-7, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26272641

RESUMO

PURPOSE: To assess the effect of 950 MHz ultra-high-frequency electromagnetic radiation (UHF-EMR) on biomarkers of oxidative damage to DNA, proteins and lipids in the left cerebral cortex (LCC) and right cerebral cortex (RCC) of neonate and 6-day-old rats. MATERIALS AND METHODS: Twelve rats were equally divided into two groups as controls (CR) and exposed (ER), for each age (0 and 6 days). The LCC and RCC were examined in ER and CR after exposure. Radiation exposure lasted 30 min per day for up to 27 days (throughout pregnancy and 6 days postnatal). The specific absorption rate ranged from 1.32-1.14 W/kg. The damage to lipids, proteins and DNA was verified by thiobarbituric acid reactive substances, carbonylated proteins (CP) and comets, respectively. The concentration of glucose in the peripheral blood of the rats was measured by the Accu-Chek Active Kit due to increased CP in RCC. RESULTS: In neonates, no modification of the biomarkers tested was detected. On the other hand, there was an increase in the levels of CP in the RCC of the 6-day-old ER. Interestingly, the concentration of blood glucose was decreased in this group. CONCLUSIONS: Our results indicate that there is no genotoxicity and oxidative stress in neonates and 6 days rats. However, the RCC had the highest concentration of CP that do not seem to be a consequence of oxidative stress. This study is the first to demonstrate the use of UHF-EMR causes different damage responses to proteins in the LCC and RCC.


Assuntos
Envelhecimento/metabolismo , Córtex Cerebral/metabolismo , Córtex Cerebral/efeitos da radiação , Proteínas do Tecido Nervoso/metabolismo , Exposição à Radiação , Espécies Reativas de Oxigênio/metabolismo , Animais , Relação Dose-Resposta a Droga , Campos Eletromagnéticos , Feminino , Masculino , Micro-Ondas , Estresse Oxidativo/fisiologia , Estresse Oxidativo/efeitos da radiação , Doses de Radiação , Ratos
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