RESUMEN
Glioblastomas are tumors that present a high mortality rate. Artemether (ART) is a lactone with antitumor properties, demonstrating low bioavailability and water solubility. In the present study, we developed lipid-core nanocapsules (LNC) containing pequi oil (Caryocar brasiliense Cambess) as the oily core for ART-loaded LNCs (LNCART) and evaluated their effect on human glioblastoma cells (U-87 MG). LNCs were developed by interfacial deposition of a preformed polymer, followed by physicochemical characterization. LNCART revealed a diameter of 0.216 µm, polydispersity index of 0.161, zeta potential of -12.0 mV, and a pH of 5.53. Furthermore, mitochondrial viability, proliferation, total antioxidant status, and antioxidant enzyme activity were evaluated. ART reduced cell viability after 24 h and proliferation after 48 h of treatment at concentrations equal to or above 40 µg . mL-1. LNCART, at 1.25 µg . mL-1, reduced these parameters after 24 h of treatment. Furthermore, superoxide dismutase (SOD) activity was elevated, while glutathione reductase (GR) activity was reduced. These findings suggest that ART loaded into LNC may be a promising alternative to improve its pharmacological action and possible application as a therapeutic agent for glioblastoma.
Asunto(s)
Glioblastoma , Nanocápsulas , Humanos , Nanocápsulas/química , Antioxidantes/farmacología , Arteméter , Supervivencia Celular , Glioblastoma/tratamiento farmacológico , Lípidos/química , PolímerosRESUMEN
The Buccal Micronucleus Cytome Assay (BMCyt) has become an important biomonitoring tool for assessing cytogenetic damage in many studied populations. Each laboratory applies protocols that vary according to the method of collecting and preparing samples. Besides, Brazil is a country of great territorial extensions that received immigrants from various parts of the world with different genetic backgrounds. Therefore, the present study aimed to evaluate the inter-laboratory variation in scoring the same set of slides using the more comprehensive scoring criteria, to standardize the BMCyt protocol, to observe the basal alterations in populations of different Brazilian regions and to compare it with other places around the world. Our results showed that a valuable number of laboratories participated, ten laboratories from different regions of the country, for the validation of the BMCyt in human biomonitoring studies, resulting in the 804 healthy individuals. This was possible because we observed: a range of measures needs to be considered, such as the baseline frequency of DNA damage and cell death in non-exposed individuals; age when grouped showed an influence on DNA damage, although when evaluated by group we did not see an influence; association between smoking habit and all endpoints of the BMCyt (except karyolytic cells) was evident; the basal MN frequency, in the majority of groups, follows those around the world; and the BMCyt was confirmed as a good health status biomarker. We emphasize the need for constant discussions on the parameters of cell death due to greater difficulty among the analyzers.
Asunto(s)
Bioensayo/normas , Núcleo Celular/genética , Células Epiteliales/ultraestructura , Laboratorios/normas , Micronúcleos con Defecto Cromosómico , Pruebas de Micronúcleos/normas , Mucosa Bucal/citología , Adolescente , Adulto , Bioensayo/métodos , Brasil , Muerte Celular/genética , Núcleo Celular/ultraestructura , Daño del ADN , Femenino , Humanos , Masculino , Micronúcleos con Defecto Cromosómico/estadística & datos numéricos , Pruebas de Micronúcleos/métodos , Persona de Mediana Edad , Mucosa Bucal/ultraestructura , Adulto JovenRESUMEN
Paullinia cupana is associated with a diverse community of pathogenic and endophytic microorganisms. We isolated and identified endophytic fungal communities from the roots and seeds of P. cupana genotypes susceptible and tolerant to anthracnose that grow in two sites of the Brazilian Amazonia forest. We assessed the antibacterial, antitumor and genotoxic activity in vitro of compounds isolated from the strains Trichoderma asperellum (1BDA) and Diaporthe phaseolorum (8S). In concert, we identified eight fungal species not previously reported as endophytes; some fungal species capable of inhibiting pathogen growth; and the production of antibiotics and compounds with bacteriostatic activity against Pseudomonas aeruginosa in both susceptible and multiresistant host strains. The plant genotype, geographic location and specially the organ influenced the composition of P. cupana endophytic fungal community. Together, our findings identify important functional roles of endophytic species found within the microbiome of P. cupana. This hypothesis requires experimental validation to propose management of this microbiome with the objective of promoting plant growth and protection.
Asunto(s)
Biodiversidad , Endófitos , Hongos/clasificación , Hongos/metabolismo , Paullinia/microbiología , Metabolismo Secundario , Animales , Células CHO , Análisis por Conglomerados , Cricetulus , Hongos/aislamiento & purificación , Metaboloma , Metabolómica/métodos , Raíces de Plantas/microbiología , Carácter Cuantitativo Heredable , Semillas/microbiologíaRESUMEN
Titanium dioxide nanocrystals (TiO2 NCs) crystalline structures include anatase, rutile and brookite. This study evaluated the genotoxic effects of 3.4 and 6.2 nm anatase TiO2 NCs and 78.0 nm predominantly rutile TiO2 NCs through an in vitro micronucleus (MN) assay using V79 cells and an in vivo somatic mutation and recombination test in Drosophila wings. The MN assay was performed with nontoxic concentrations of TiO2 NCs. Only anatase (3.4 nm) at the highest concentration (120 µM) induced genotoxicity in V79 cells. In the in vivo test, Drosophila melanogaster larvae obtained from standard (ST) or high bioactivation (HB) crosses were treated with TiO2 NCs. In the ST cross, no mutagenic effects were observed. However, in the HB cross, TiO2 NCs (3.4 nm) were mutagenic at 1.5625 and 3.125 mM, while 78.0 nm NCs increased mutant spots at all concentrations tested except 3.125 mM. Only the smallest anatase TiO2 NCs induced mutagenic effects in vitro and in vivo. For rutile TiO2 NCs, no clastogenic/aneugenic effects were observed in the MN assay. However, they were mutagenic in Drosophila. Therefore, both anatase and rutile TiO2 NCs induced mutagenicity. Further research is necessary to clarify the TiO2 NCs genotoxic/mutagenic action mechanisms.
Asunto(s)
Citocinesis , Daño del ADN/efectos de los fármacos , Drosophila melanogaster/efectos de los fármacos , Nanopartículas del Metal/toxicidad , Pruebas de Micronúcleos/métodos , Titanio/toxicidad , Alas de Animales/efectos de los fármacos , Animales , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Cricetulus , Drosophila melanogaster/citología , Drosophila melanogaster/genética , Femenino , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Larva/citología , Larva/efectos de los fármacos , Larva/genética , Pulmón/citología , Pulmón/efectos de los fármacos , Masculino , Pruebas de MutagenicidadRESUMEN
In this study, we evaluated the toxic and genotoxic potential of zinc oxide nanoparticles (ZnO NPs) of 20 nm and the mutagenic potential of these ZnO NPs as well as that of an amorphous ZnO. Toxicity was assessed by XTT colorimetric assay. ZnO NPs were toxic at concentrations equal to or higher than 240.0 µM. Genotoxicity was assessed by in vitro Cytokinesis Block Micronucleus Assay (CBMN) in V79 cells. ZnO NPs were genotoxic at 120.0 µM. The mutagenic potential of amorphous ZnO and the ZnO NPs was assayed using the wing Somatic Mutation and Recombination Test (SMART) of Drosophila melanogaster. In the Standard cross, the amorphous ZnO and ZnO NPs were not mutagenic. Nevertheless, Marker trans-heterozygous individuals from the High bioactivation cross treated with amorphous ZnO (6.25 mM) and ZnO NPs (12.50 mM) displayed a significant increased number of mutant spots when compared with the negative control. In conclusion, the results were not dose related and indicate that only higher concentrations of ZnO NPs were toxic and able to induce genotoxicity in V79 cells. The increase in mutant spots observed in D. melanogaster was generated due to mitotic recombination, rather than mutational events.
Asunto(s)
Nanopartículas del Metal/toxicidad , Mutagénesis/efectos de los fármacos , Mutágenos/toxicidad , Recombinación Genética/efectos de los fármacos , Óxido de Zinc/toxicidad , Animales , Animales Modificados Genéticamente , Bioensayo , Línea Celular , Cricetulus , Cruzamientos Genéticos , Drosophila melanogaster , Femenino , Marcadores Genéticos/efectos de los fármacos , Pérdida de Heterocigocidad/efectos de los fármacos , Masculino , Nanopartículas del Metal/ultraestructura , Pruebas de Micronúcleos , Pruebas de Mutagenicidad , Mutágenos/química , Pigmentación/efectos de los fármacos , Alas de Animales , Óxido de Zinc/químicaRESUMEN
Leprosy transmission still occurs despite the availability of highly effective treatment. The next step towards successfully eliminating leprosy is interrupting the chain of transmission of the aetiological agent, Mycobacterium leprae. In this investigation, we provide evidence that household contacts (HHCs) of leprosy patients might not only have subclinical infections, but may also be actively involved in bacilli transmission. We studied 444 patients and 1,352 contacts using anti-phenolic glycolipid-I (PGL-I) serology and quantitative polymerase chain reaction (qPCR) to test for M. leprae DNA in nasal swabs. We classified the patients according to the clinical form of their disease and the contacts according to the characteristics of their index case. Overall, 63.3% and 34.2% of patients tested positive by ELISA and PCR, respectively. For HHCs, 13.3% had a positive ELISA test result and 4.7% had a positive PCR test result. The presence of circulating anti-PGL-I among healthy contacts (with or without a positive PCR test result from nasal swabs) was considered to indicate a subclinical infection. DNA detected in nasal swabs also indicates the presence of bacilli at the site of transmission and bacterial entrance. We suggest that the concomitant use of both assays may allow us to detect subclinical infection in HHCs and to identify possible bacilli carriers who may transmit and disseminate disease in endemic regions. Chemoprophylaxis of these contacts is suggested.
Asunto(s)
Humanos , Antígenos Bacterianos/sangre , Composición Familiar , Glucolípidos/sangre , Lepra/transmisión , Mycobacterium leprae , Infecciones Asintomáticas , Anticuerpos Antibacterianos/sangre , Portador Sano , ADN Bacteriano/análisis , Lepra/diagnóstico , Lepra/epidemiología , Mycobacterium leprae/genética , Mycobacterium leprae/inmunología , Mucosa Nasal/microbiología , Reacción en Cadena de la Polimerasa , PrevalenciaRESUMEN
Leprosy transmission still occurs despite the availability of highly effective treatment. The next step towards successfully eliminating leprosy is interrupting the chain of transmission of the aetiological agent, Mycobacterium leprae. In this investigation, we provide evidence that household contacts (HHCs) of leprosy patients might not only have subclinical infections, but may also be actively involved in bacilli transmission. We studied 444 patients and 1,352 contacts using anti-phenolic glycolipid-I (PGL-I) serology and quantitative polymerase chain reaction (qPCR) to test for M. leprae DNA in nasal swabs. We classified the patients according to the clinical form of their disease and the contacts according to the characteristics of their index case. Overall, 63.3% and 34.2% of patients tested positive by ELISA and PCR, respectively. For HHCs, 13.3% had a positive ELISA test result and 4.7% had a positive PCR test result. The presence of circulating anti-PGL-I among healthy contacts (with or without a positive PCR test result from nasal swabs) was considered to indicate a subclinical infection. DNA detected in nasal swabs also indicates the presence of bacilli at the site of transmission and bacterial entrance. We suggest that the concomitant use of both assays may allow us to detect subclinical infection in HHCs and to identify possible bacilli carriers who may transmit and disseminate disease in endemic regions. Chemoprophylaxis of these contacts is suggested.
Asunto(s)
Antígenos Bacterianos/sangre , Composición Familiar , Glucolípidos/sangre , Lepra/transmisión , Mycobacterium leprae , Anticuerpos Antibacterianos/sangre , Infecciones Asintomáticas , Portador Sano , ADN Bacteriano/análisis , Humanos , Lepra/diagnóstico , Lepra/epidemiología , Mycobacterium leprae/genética , Mycobacterium leprae/inmunología , Mucosa Nasal/microbiología , Reacción en Cadena de la Polimerasa , PrevalenciaRESUMEN
OBJECTIVE: Our aim was to compare the performance of three serological assays in leprosy patients and their household contacts utilising two quantitative ELISA tests using native PGL-I (PGL-1 ELISA), synthetic ND-O-HSA (ND-O-HSA ELISA), and the semi-quantitative lateral flow test (ML Flow). METHODS: Comparisons among three immunological assays, PGL-I ELISA, ND-O-HSA ELISA, and ML Flow were performed in 154 leprosy patients, 191 household contacts and 52 health subjects. RESULTS: The sensitivity results of the PGL-1, ND-O-HSA, and ML Flow were 68.83%, 63.84%, and 60.65%, respectively, with specificity of 98% for both ELISA assays. The native and synthetic PGL-I ELISA assays detected antibodies in 22.73% and 31.82% of the paucibacillary (PB) patients, respectively and the ML Flow test did not detect antibodies in this group. The ML Flow test was able to discriminate patients into PB or multibacillary (MB) forms, while the native PGL-I and ND-O-HSA was correlated with the bacillary load and the Ridley-Jopling clinical forms. In household contacts, the native PGL-I, ND-O-HSA, and ML Flow assays detected seropositivity of 25%, 17%, and 10%, respectively. CONCLUSIONS: The use of ELISA and ML Flow tests are thus recommended as additional tools in the diagnosis and classification of the clinical forms, aiding in prescribing the correct treatment regimen to prevent subsequent nerve damage and disability, and besides, the PGL-I ELISA may be used to detect subclinical infection in leprosy.