ABSTRACT
This study evaluated the genotoxicity of lyophilized glycolic extract of Theobroma cacao Linné seeds (TCL), using the micronucleus assay in bone marrow of mice. The interaction between TCL and doxorubicin (DXR) was also analyzed. Experimental groups were evaluated 24-48 h after treatment with N-Nitroso-N-ethylurea (NEU: 50 mg/kg), DXR (5 mg/kg), NaCl (145 mM), TCL (0.5-2 g/kg), and TCL (2 g/kg) in combination with DXR (antigenotoxic assays). Analysis of micronucleated polychromatic erythrocytes (MNPCEs) showed no significant differences between all the treatment doses of TCL and NaCl control. Mice experimentally treated with DXR and NEU significantly induced MNPCEs. However, a significant reduction of MNPCEs was also observed when TCL was administered in combination with the chemotherapeutic agent DXR. The analysis of the PCE/NCE ratio revealed no significant differences between the NaCl control, all doses of TCL, and DXR. However, there were significant differences in the PCE/NCE ratio between positive NEU control and all other treatments. The PCE/NCE ratio observed after treatment with TCL and DXR showed significant differences and intermediate values to controls (NaCl and NEU). This study suggests absence of genotoxicity and cytotoxicity of TCL, regardless of dose, sex, and time. TCL reduced genotoxic effects induced by DXR, suggesting potential antigenotoxic effects.
Subject(s)
Cacao , Animals , DNA Damage , Doxorubicin , Erythrocytes , Mice , Micronucleus Tests , Plant Extracts/pharmacologyABSTRACT
This study evaluated the genotoxicity of lyophilized glycolic extract of Theobroma cacao Linné seeds (TCL), using the micronucleus assay in bone marrow of mice. The interaction between TCL and doxorubicin (DXR) was also analyzed. Experimental groups were evaluated 24-48 h after treatment with N-Nitroso-N-ethylurea (NEU: 50 mg/kg), DXR (5 mg/kg), NaCl (145 mM), TCL (0.5-2 g/kg), and TCL (2 g/kg) in combination with DXR (antigenotoxic assays). Analysis of micronucleated polychromatic erythrocytes (MNPCEs) showed no significant differences between all the treatment doses of TCL and NaCl control. Mice experimentally treated with DXR and NEU significantly induced MNPCEs. However, a significant reduction of MNPCEs was also observed when TCL was administered in combination with the chemotherapeutic agent DXR. The analysis of the PCE/NCE ratio revealed no significant differences between the NaCl control, all doses of TCL, and DXR. However, there were significant differences in the PCE/NCE ratio between positive NEU control and all other treatments. The PCE/NCE ratio observed after treatment with TCL and DXR showed significant differences and intermediate values to controls (NaCl and NEU). This study suggests absence of genotoxicity and cytotoxicity of TCL, regardless of dose, sex, and time. TCL reduced genotoxic effects induced by DXR, suggesting potential antigenotoxic effects.
Este estudo avaliou a genotoxicidade do extrato glicólico liofilizado de sementes de Theobroma cacao Linné (TCL), usando o ensaio do micronúcleo em medula óssea de camundongos. A interação entre TCL e doxorrubicina (DXR) foi também analisada. Grupos experimentais foram avaliados 24-48 h após tratamento com N-Nitroso-N-etilureia (NEU: 50 mg/kg), DXR (5 mg/kg), NaCl (145 mM), TCL (0,5-2 g/kg), e TCL (2 g/kg) em combinação com DXR (ensaio antigenotóxico). As análises de eritrócitos policromáticos micronucleados (EPCMNs) não mostraram diferenças significantes entre todas as doses de tratamento do TCL e o controle NaCl. Camundongos experimentalmente tratados com DXR e NEU induziram significativamente EPCMNs. Contudo, uma redução significante de EPCMNs foi também observada quando TCL foi administrada em combinação com o agente quimioterapêutico DXR. As análises da relação EPC/ENC (eritrócito policromático/eritrócito normocromático) revelaram ausência de diferenças significantes entre o controle NaCl, todas as doses de TCL e DXR. Contudo, houve diferenças significantes na relação EPC/ENC entre o controle positivo NEU e todos os outros tratamento. A relação ECP/ENC observada após o tratamento com TCL e DXR mostrou diferenças significantes e valores intermediários aos controles (NaCl e NEU). Este estudo sugere ausência de genotoxicidade e citotoxicidade de TCL, independentemente da dose, sexo e tempo. TCL reduziu os efeitos genotóxicos induzidos por DXR, sugerindo potencial efeitos antigenotóxicos.
Subject(s)
Animals , Mice , DNA Damage , Cacao/toxicity , Cytotoxins/analysis , Plant Extracts/pharmacology , Micronucleus Tests , Doxorubicin , ErythrocytesABSTRACT
Abstract Handroanthus impetiginosus has long been used in traditional medicine and various studies have determined the presence of bioactive chemical compounds and potential phytotherapeutics. In this study, the genotoxicity of the lyophilized tincture of H. impetiginosus bark (THI) was evaluated in mouse bone marrow using micronucleus assays. The interaction between THI and genotoxic effects induced by the chemotherapeutic agent, doxorubicin (DXR), was also analyzed. Experimental groups were evaluated 24 to 48 h after treatment with N-nitroso-N-ethylurea (NEU; 50 mg/kg), DXR (5 mg/kg), sodium chloride (NaCl; 150 mM), and THI (0.5-2 g/kg). Antigenotoxic assays were carried out using THI (0.5 g/kg) in combination with NEU or DXR. Analysis of the micronucleated polychromatic erythrocytes (MNPCEs) indicated no significant differences between treatment doses of THI (0.5-2 g/kg) and NaCl. Polychromatic erythrocyte (PCE) to normochromatic erythrocyte (NCE) ratios did not indicate any statistical differences between DXR and THI or NaCl, but there were differences between THI and NaCl. A significant reduction in MNPCEs and PCE/NCE ratios was observed when THI was administered in combination with DXR. This study suggested the absence of THI genotoxicity that was dose-, time-, and gender-independent and the presence of moderate systemic toxicity that was dose-independent, but time- and gender-dependent. The combination of THI and DXR also suggested antigenotoxic effects, indicating that THI reduced genotoxic effects induced by chemotherapeutic agents.
Resumo Handroanthus impetiginosus tem sido usada durante um longo período pela medicina tradicional e vários estudos têm demonstrados a presença de compostos químicos e potencial fitoterapêutico. Esta pesquisa avaliou a genotoxicidade da tintura da casca liofilizada de H. impetiginosus (THI) usando o ensaio do micronúcleo em medula óssea de camundongos. A interação entre THI e os efeitos genotóxicos induzidos pelo quimioterápico doxorrubicina (DXR) também foram analisados. Grupos experimentais foram analisados a 24-48 h após o tratamento com N-Nitroso-N-etiluréia (NEU; 50 mg/kg), DXR (5 mg/kg), NaCl (150 mM) e THI (0,5-2 g/kg). O ensaio antigenotóxico foi conduzido utilizando THI (0,5 g/kg) em combinação com NEU ou DXR. A análise de eritrócitos policromáticos micronucleados (EPCMNs) não mostrou diferenças significativas entre as doses de tratamento (0,5-2 g/kg) e NaCl. As proporções de eritrócitos policromáticos (EPC)/eritrócitos normocromáticos (ENC) não revelaram diferenças estatísticas entre DXR e THI ou NaCl, porém houve diferenças entre THI e NaCl. Uma redução significativa em EPCMNs e na razão EPC/ENC foi observada quando THI foi administrado em combinação com DXR. Essa pesquisa sugere ausência de genotoxicidade de THI, dose-, tempo- e sexo-independente, e moderada toxicidade sistêmica dose-independente, mas tempo- e sexo-dependente. A associação do THI e DXR também sugere efeitos antigenotóxicos. Por conseguinte, THI pode reduzir os efeitos genotóxicos induzidos pelo quimioterapêutico.
Subject(s)
Animals , Mice , DNA Damage/drug effects , Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Plant Extracts/pharmacology , Doxorubicin/toxicity , Protective Agents/pharmacology , Micronucleus Tests , Cells, Cultured , Tabebuia/chemistryABSTRACT
Handroanthus impetiginosus has long been used in traditional medicine and various studies have determined the presence of bioactive chemical compounds and potential phytotherapeutics. In this study, the genotoxicity of the lyophilized tincture of H. impetiginosus bark (THI) was evaluated in mouse bone marrow using micronucleus assays. The interaction between THI and genotoxic effects induced by the chemotherapeutic agent, doxorubicin (DXR), was also analyzed. Experimental groups were evaluated 24 to 48 h after treatment with N-nitroso-N-ethylurea (NEU; 50 mg/kg), DXR (5 mg/kg), sodium chloride (NaCl; 150 mM), and THI (0.5-2 g/kg). Antigenotoxic assays were carried out using THI (0.5 g/kg) in combination with NEU or DXR. Analysis of the micronucleated polychromatic erythrocytes (MNPCEs) indicated no significant differences between treatment doses of THI (0.5-2 g/kg) and NaCl. Polychromatic erythrocyte (PCE) to normochromatic erythrocyte (NCE) ratios did not indicate any statistical differences between DXR and THI or NaCl, but there were differences between THI and NaCl. A significant reduction in MNPCEs and PCE/NCE ratios was observed when THI was administered in combination with DXR. This study suggested the absence of THI genotoxicity that was dose-, time-, and gender-independent and the presence of moderate systemic toxicity that was dose-independent, but time- and gender-dependent. The combination of THI and DXR also suggested antigenotoxic effects, indicating that THI reduced genotoxic effects induced by chemotherapeutic agents.
Subject(s)
Bone Marrow Cells , DNA Damage/drug effects , Doxorubicin/toxicity , Plant Extracts/pharmacology , Protective Agents/pharmacology , Tabebuia/chemistry , Animals , Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Cells, Cultured , Mice , Micronucleus TestsABSTRACT
Abstract This study evaluated the genotoxicity of lyophilized glycolic extract of Theobroma cacao Linné seeds (TCL), using the micronucleus assay in bone marrow of mice. The interaction between TCL and doxorubicin (DXR) was also analyzed. Experimental groups were evaluated 24-48 h after treatment with N-Nitroso-N-ethylurea (NEU: 50 mg/kg), DXR (5 mg/kg), NaCl (145 mM), TCL (0.5-2 g/kg), and TCL (2 g/kg) in combination with DXR (antigenotoxic assays). Analysis of micronucleated polychromatic erythrocytes (MNPCEs) showed no significant differences between all the treatment doses of TCL and NaCl control. Mice experimentally treated with DXR and NEU significantly induced MNPCEs. However, a significant reduction of MNPCEs was also observed when TCL was administered in combination with the chemotherapeutic agent DXR. The analysis of the PCE/NCE ratio revealed no significant differences between the NaCl control, all doses of TCL, and DXR. However, there were significant differences in the PCE/NCE ratio between positive NEU control and all other treatments. The PCE/NCE ratio observed after treatment with TCL and DXR showed significant differences and intermediate values to controls (NaCl and NEU). This study suggests absence of genotoxicity and cytotoxicity of TCL, regardless of dose, sex, and time. TCL reduced genotoxic effects induced by DXR, suggesting potential antigenotoxic effects.
Resumo Este estudo avaliou a genotoxicidade do extrato glicólico liofilizado de sementes de Theobroma cacao Linné (TCL), usando o ensaio do micronúcleo em medula óssea de camundongos. A interação entre TCL e doxorrubicina (DXR) foi também analisada. Grupos experimentais foram avaliados 24-48 h após tratamento com N-Nitroso-N-etilureia (NEU: 50 mg/kg), DXR (5 mg/kg), NaCl (145 mM), TCL (0,5-2 g/kg), e TCL (2 g/kg) em combinação com DXR (ensaio antigenotóxico). As análises de eritrócitos policromáticos micronucleados (EPCMNs) não mostraram diferenças significantes entre todas as doses de tratamento do TCL e o controle NaCl. Camundongos experimentalmente tratados com DXR e NEU induziram significativamente EPCMNs. Contudo, uma redução significante de EPCMNs foi também observada quando TCL foi administrada em combinação com o agente quimioterapêutico DXR. As análises da relação EPC/ENC (eritrócito policromático/eritrócito normocromático) revelaram ausência de diferenças significantes entre o controle NaCl, todas as doses de TCL e DXR. Contudo, houve diferenças significantes na relação EPC/ENC entre o controle positivo NEU e todos os outros tratamento. A relação ECP/ENC observada após o tratamento com TCL e DXR mostrou diferenças significantes e valores intermediários aos controles (NaCl e NEU). Este estudo sugere ausência de genotoxicidade e citotoxicidade de TCL, independentemente da dose, sexo e tempo. TCL reduziu os efeitos genotóxicos induzidos por DXR, sugerindo potencial efeitos antigenotóxicos.
ABSTRACT
AIMS: The present study verified the effect of propolis alone and its association with swimming in dyslipidemia, left ventricular hypertrophy and atherogenesis of hypercholesterolemic mice. METHODS AND RESULTS: The experiments were performed in LDLr-/- mice, fed with high fat diet for 75 days, and were divided into four experimental groups (n=10): HL, sedentary, subjected to aquatic stress (5 min per day, 5 times per week); NAT submitted to a swimming protocol (1 hour per day, 5 times per week) from the 16th day of the experiment; PRO, sedentary, submitted to aquatic stress and which received oral propolis extract (70 uL/animal/day) from the 16th day of the experiment; HL+NAT+PRO, submitted to swimming and which received propolis as described above. After 75 days, blood was collected for analysis of serum lipids. The ratio between the ventricular weight (mg) and the animal weight (g) was calculated. Histological sections of the heart and aorta were processed immunohistochemically with anti-CD40L antibodies to evaluate the inflammatory process; stained with hematoxylin/eosin and picrosirius red to assess morphological and morphometric alterations. The HL animals showed severe dyslipidemia, atherogenesis and left ventricular hypertrophy, associated with a decrease in serum HDLc levels and subsequent development of cardiovascular inflammatory process, characterized by increased expression of CD40L in the left ventricle and aorta. Swimming and propolis alone and\or associated prevented the LVH, atherogenesis and arterial and ventricular inflammation, decreasing the CD40L expression and increasing the HDLc plasmatic levels. CONCLUSION: Propolis alone or associated with a regular physical activity is beneficial in cardiovascular protection through anti-inflammatory action.
Subject(s)
Atherosclerosis/prevention & control , Hypercholesterolemia/complications , Hypertrophy, Left Ventricular/prevention & control , Physical Conditioning, Animal , Propolis/administration & dosage , Swimming , Animals , Atherosclerosis/etiology , Hypertrophy, Left Ventricular/etiology , Male , Mice , Mice, Inbred C57BLABSTRACT
Inúmeros esforços têm sido dirigidos para conferir às plantas seu real papel e valor na terapia. Este estudo teve como objetivo avaliar a atividade antimicrobiana, mutagênica, toxicidade, e os efeitos no crescimento e diferenciação de Herpetomonas samuelpessoai, do extrato hidroalcoólico de Bauhinia forficata. Para avaliar a atividade antimicrobiana foi realizado o teste de difusão em ágar, bem como a determinação das concentrações inibitória (CIM) e microbicida mínimas (CMM). O potencial clastogênico e/ou aneugênico, in vivo, foi avaliado usando o teste do micronúcleo em medula óssea de camundongos Swiss albinus. Foi determinada também a dose letal média (DL50). O extrato inibiu o crescimento de oito bactérias, mostrando-se mais ativo para Gram-positivas e não foi eficiente para os fungos, tendo sido ativo nas concentrações de 2000, 1000, 500 e 250 mg/mL contra os microrganismos testados. Os resultados mostraram que nas concentrações administradas (500, 1000 e 2000 mg/Kg), não houve aumento estatisticamente significativo de micronúcleos. Não houve ação no crescimento e diferenciação de Herpetomonas samuelpessoai nas concentrações testadas. Com relação a DL50, o extrato não apresentou toxicidade.
Numerous efforts have been directed to discover the role and the value of plants in therapy. This work aimed to evaluate the antimicrobial activity, mutagenicity, toxicity and effects on growth and differentiation of Herpetomonas samuelpessoai of the hydroalcoholic extract of Bauhinia forficata. To evaluate the antimicrobial activity it was performed the agar diffusion test, minimum inhibitory (MIC) and microbicidal (MMC) concentrations. The in vivo clastogenic and / or aneugênic potential was evaluated using the micronucleus test in mice bone marrow Swiss albinus. It was also determined the median lethal dose (LD50). The extract inhibited the growth of eight bacteria, being more active against Gram-positiveones, and was not active against fungi. The microorganisms tested had MIC concentrations of 2000, 1000, 500 and 250 mg / mL. The results showed that the tested concentrations (500, 1000 and 2000 mg / kg) had no statistically significant increasedthe micronucleus. There was no action on the growth and differentiation of Herpetomonas samuelpessoai at the concentrations tested. In respect to the LD50, the extract showed no toxicity.
Subject(s)
Animals , Male , Female , Mice , Trypanosomatina/growth & development , Bauhinia/drug effects , Anti-Infective Agents/pharmacology , Mutagenicity TestsABSTRACT
INTRODUCTION: The aim of this study was to investigate the oral colonization profile of Candida albicans strains isolated from diabetic patients and their non-diabetic consorts. In addition hydrolytic enzyme activity of these isolates was analysed. METHODS: The genetic diversity of C. albicans oral isolates from 52 couples was established using isoenzyme marker and cluster analysis. Hydrolytic enzyme characteristics, namely secreted aspartyl proteinases (SAPs) and phospholipases (PLs) were also analysed. RESULTS: Simultaneous colonization by C. albicans was observed in the consorts of 12 couples (23.1%). Patterns of monoclonal and polyclonal oral colonization by C. albicans strains were identified and the coexistence of identical or highly related strains was observed in both members of eight couples. The genetic diversity observed in the total yeast population revealed four large, genetically distinct groups (A to D) and the coexistence of strains in couples or consorts conjugally unrelated. SAP and PL activity was observed in the majority of C. albicans isolates without any association to particular strain, strain clusters (highly related isolates), or clinical characteristics of the consorts (diabetic, non-diabetic, and gender). CONCLUSION: Possible sources of transmission and oral propagation of groups (clusters) of strains of C. albicans can occur between diabetic and non-diabetic consorts. A conjugal genotypic identity exists in most C. albicans-positive couples, that is, both consorts share identical or highly related strains; however, this identity is not couple-specific as seen by the coexistence of clusters in couples and unrelated consorts.
Subject(s)
Aspartic Acid Proteases/metabolism , Candida albicans/enzymology , Candida albicans/genetics , Candidiasis, Oral/microbiology , Diabetes Complications , Phospholipases/metabolism , Adult , Aged , Alleles , Candidiasis, Oral/complications , Candidiasis, Oral/transmission , Cluster Analysis , Disease Transmission, Infectious , Electrophoresis/methods , Female , Genetic Variation , Humans , Male , Middle Aged , Species Specificity , Spouses , VirulenceABSTRACT
Cinco cepas de Candida albicans y otras cinco de C. dubliniensis así como sus cepas-patrón fueron evaluadas mediante el empleo de técnicas de electroforesis en geles de almidón y de poliacrilamida. Después de las electroforesis y evaluación numérica, se obtuvieron dos taxa muy distintos, que pueden justificar el uso de MLEE y SDS-PAGE como métodos confiables para la diferenciación e identificación complementaria de C.dubliniensis.
Subject(s)
Candida albicans , Electrophoresis, Polyacrylamide Gel , Electrophoresis, Starch Gel , BrazilABSTRACT
Cinco cepas de Candida albicans y otras cinco de C. dubliniensis así como sus cepas-patrón fueron evaluadas mediante el empleo de técnicas de electroforesis en geles de almidón y de poliacrilamida. Después de las electroforesis y evaluación numérica, se obtuvieron dos taxa muy distintos, que pueden justificar el uso de MLEE y SDS-PAGE como métodos confiables para la diferenciación e identificación complementaria de C.dubliniensis. (AU)
Subject(s)
Candida albicans , Electrophoresis, Starch Gel , Electrophoresis, Polyacrylamide Gel , BrazilABSTRACT
The application of gel electrophoresis and numerical analysis of yeast soluble proteins analysis to the investigation of 12 oral yeast strains belonging to five species is described. It involves one-dimensional electrophoresis of SDS-solubilized whole-cell proteins using different culture media for the cultivation of the cells, integration densitometries in the areas of the gels and percentages of the proteins extraction. These extracts were prepared from four isolates of Candida albicans, two of C. tropicalis, C. guilliermondii, C. parapsilosis and C. krusei. The extracts from whole-cells proteins using different culture media for the cultivation of the cells were fractionated by slab electrophoresis using a discontinuous buffer system. The corresponding patterns showed at least 36 polypeptides in the range of 14.4-200 kDa. Different isolates of each species were clearly different in each of the five species. The data obtained suggest that different nutritional compositions led to the expression of different proteins derived from alternatives metabolic pathways expressed by the electrophoretic profiles. The construction of a database of protein fingerprints and numerical analysis based on such data, may have some implications in the classification and identification of such species with epidemiological, ecological and taxonomic purposes. A well defined or synthetic culture media seems to be much properly
Subject(s)
Humans , Candida , Culture Media , Mouth , Fungal Proteins/analysis , Candida , Electrophoresis, Polyacrylamide Gel , Numerical Analysis, Computer-AssistedABSTRACT
Electrophoretic studies of multilocus-enzymes (MLEE) and whole-cell protein (SDS-PAGE) were carried out in order to evaluate the parity between different methods for the characterization of five Candida species commonly isolated from oral cavity of humans by numerical taxonomy methods. The obtained data revealed that sodium dodecyl sulfate polyacrylamide gel electrophoresis is more efficient in grouping strains in their respective species while MLEE has much limited resolution in organizing all strains in their respective species-specific clusters. MLEE technique must be regarded for surveys in which just one species of Candida is involved
Subject(s)
Humans , Candida/chemistry , Electrophoresis/methods , Enzymes/analysis , Mouth/microbiology , Proteins/analysis , Candida/classification , Electrophoresis, Polyacrylamide Gel/methodsABSTRACT
Even in normalized gels, some problems with the choice and position of protein bands always have hindered the processing of electrophoretic data. We have developed a way to establish which criterion best fits the necessities in order to maximize the similarity indexes for numerical analysis. Some repetitions of a Candida albicans strain were carried out in eleven different gels. After staining, the bands were scored in numbers within ranges of +/- n values with increases of one quarter steps (+/- 0.25 kDa, +/- 0.5 kDa, +/- 0.75 kDa, +/- 1.0 kDa, +/- 1.25 kDa) that will limit the possibility of variation for the same bands that will appear on the repetitions. Using this criterion, we have determined that values scored within +/- 1.25 kDa could optimize the minimum limiting value of similarity in different dendrograms built.
Subject(s)
Candida albicans , Classification/methods , Densitometry , Electrophoresis, Polyacrylamide Gel , Mycology , Fungal Proteins/analysis , Candida albicans , Confidence Intervals , Species Specificity , Molecular WeightABSTRACT
Even in normalized gels, some problems with the choice and position of protein bands always have hindered the processing of electrophoretic data. We have developed a way to establish which criterion best fits the necessities in order to maximize the similarity indexes for numerical analysis. Some repetitions of a Candida albicans strain were carried out in eleven different gels. After staining, the bands were scored in numbers within ranges of +/- n values with increases of one quarter steps (+/- 0.25 kDa, +/- 0.5 kDa, +/- 0.75 kDa, +/- 1.0 kDa, +/- 1.25 kDa) that will limit the possibility of variation for the same bands that will appear on the repetitions. Using this criterion, we have determined that values scored within +/- 1.25 kDa could optimize the minimum limiting value of similarity in different dendrograms built.(AU)