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1.
Acta Trop ; 254: 107195, 2024 Mar 27.
Artículo en Inglés | MEDLINE | ID: mdl-38548212

RESUMEN

A clinical case of an adult horse with invasive, ulcerative, proliferative, pyogranulomatous disease of the skin (tumor) in the shoulder region is presented. The mass had a granulomatous and crater-shaped appearance, with serosanguinous discharge and the presence of fistulas with caseous material. The tumor was removed by surgery and sent to the laboratory for diagnosis. Histopathology was performed using Grocott-Gomori methenamine silver stain. The presence of necrotic material, fibrosis, infiltrated cells, and brown-colored hyphae, characteristic of members of the genus Pythium, were observed. To identify the infecting species, conventional PCRs for the amplification of the ITS-1 was carried out. Histopathological and PCR tests confirmed infection by a Pythium insidiosum strain closely associated with previous records from the US and Central America. Our report represents the first molecularly confirmed case of equine pythiosis in Mexico.

2.
Int J Toxicol ; : 10915818231225661, 2024 Jan 06.
Artículo en Inglés | MEDLINE | ID: mdl-38183303

RESUMEN

This work investigated the safety of extracts obtained from plants growing in Colombia, which have previously shown UV-filter/antigenotoxic properties. The compounds in plant extracts obtained by the supercritical fluid (CO2) extraction method were identified using gas chromatography coupled to mass spectrometry (GC/MS) analysis. Cytotoxicity measured as cytotoxic concentration 50% (CC50) and genotoxicity of the plant extracts and some compounds were studied in human fibroblasts using the trypan blue exclusion assay and the Comet assay, respectively. The extracts from Pipper eriopodon and Salvia aratocensis species and the compound trans-ß-caryophyllene were clearly cytotoxic to human fibroblasts. Conversely, Achyrocline satureioides, Chromolaena pellia, and Lippia origanoides extracts were relatively less cytotoxic with CC50 values of 173, 184, and 89 µg/mL, respectively. The C. pellia and L. origanoides extracts produced some degree of DNA breaks at cytotoxic concentrations. The cytotoxicity of the studied compounds was as follows, with lower CC50 values representing the most cytotoxic compounds: resveratrol (91 µM) > pinocembrin (144 µM) > quercetin (222 µM) > titanium dioxide (704 µM). Quercetin was unique among the compounds assayed in being genotoxic to human fibroblasts. Our work indicates that phytochemicals can be cytotoxic and genotoxic, demonstrating the need to establish safe concentrations of these extracts for their potential use in cosmetics.

3.
FEMS Microbiol Lett ; 3702023 01 17.
Artículo en Inglés | MEDLINE | ID: mdl-37822017

RESUMEN

Actinobacteria are known to produce a variety of secondary metabolites with skin-protective properties. This study aimed to investigate the photoprotective and antigenotoxic properties against UVB of extracts obtained from Cutibacterium acnes strains. Bacterial growth was measured spectrophotometrically and the constant maximum growth rate (µ) value to each strain, were calculated. In vitro photoprotection efficacy was evaluated using in vitro indices such as sun protection factor (SPFespectrophotometric) and critical wavelength (λc). UVB-antigenotoxicity was also evaluated using the SOS Chromotest. Correlation analysis was used to examine the relationship between SPFespectrophotometric and extract concentration and the %GI estimates. Among the studied strains, one showed low (6.0 ≤ SPFespectrophotometric ≤ 14.9) and eight showed media (15.0 ≤ SPFespectrophotometric ≤ 29.9) UVB photoprotection efficacy. All of them resulted in broad-spectrum (UVA-UVB) photoprotection (λc > 370 nm). In total, two C. acnes ecotypes with different growth rates were evidenced, but the protective metabolites in the extracts were produced without the influence of growth rate. Photoprotective efficacy depended on the extract concentration and was correlated with antigenotoxicity. We demonstrated that C. acnes extracts can be used as sunscreen ingredients that reduce UVB-induced genotoxicity.


Asunto(s)
Ecotipo , Rayos Ultravioleta , Protectores Solares/farmacología , Piel , Extractos Vegetales/farmacología
4.
Vet Res Commun ; 47(4): 2145-2152, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37470941

RESUMEN

The study of lice associated with domestic cats is a neglected area of veterinary parasitology. In particular, the presence of the cat louse Felicola subrostratus, a small Ischnoceran species found in the fur of the domestic cat, is rarely recognized. In America, this species has been reported across six countries. Although it was also recently reported in Mexico, no studies on the molecular identification of the specimens or the monitoring of potential bacterial, and protozoan pathogens have been carried out. Thus, this work aimed to collect, and identify lice associated with domestic and free ranging cats from the states of Veracruz and Tabasco, using amplification and sequencing of the mitochondrial cytochrome c oxidase subunit I (COI), and the ribosomal 18S rDNA genes, and to monitor selected vector-borne bacterial (Bartonella, Mycoplasma, and Rickettsia) and protozoan (Babesia, and Hepatozoon) agents. Only entire lice were used for molecular host and pathogen identification. Eighty-one lice, identified as F. subrostratus, were recovered from five infested cats, and 30 were selected for molecular identification and pathogen surveillance. Analysis of the COI and 18S rDNA partial sequences showed a similarity of 96.79%-100% with sequences of F. subrostratus from the US. Mycoplasma haemofelis and Hepatozoon canis DNA was detected in three and four samples, respectively. This work provides new collection locations for F. subrostratus, and the first sequences of the COI and 18S rDNA genes from Mexico. It also reports two pathogenic microorganisms found in the lice.


Asunto(s)
Babesia , Enfermedades de los Gatos , Animales , Gatos , México , Babesia/genética , ADN Ribosómico
5.
Rev. colomb. biotecnol ; 24(2): 16-25, jul.-dic. 2022. tab, graf
Artículo en Español | LILACS-Express | LILACS | ID: biblio-1423771

RESUMEN

RESUMEN En el trabajo se estudió un consorcio microbiano metanogénico de una mina de carbón de la cuenca de Bogotá en Colombia. Se establecieron cultivos de enriquecimiento de carbón ex situ para el crecimiento y la producción de gas de novo. El gas biogénico producido por los cultivos se analizó mediante cromatografía de gases con detectores de ionización de llama y conductividad térmica. Los cultivos se utilizaron para aislar estirpes microbianas y para generar bibliotecas del gene 16S rARN empleando de cebadores de bacteria y de arquea. El análisis de cromatografía de gases mostró producción de metano a 37 oC, pero no a 60 oC, donde el CO2 fue el componente principal del gas biogénico. El análisis de la secuencia del gen 16S rARN de estirpes microbianos y de las bibliotecas de clones, estableció que el consorcio microbiano metanogénico estuvo formado por especies de bacterias de los géneros Bacillus y Gracilibacter más la arquea del género Methanothermobacter. El consorcio microbiano metanogénico identificado es potencialmente responsable de la generación de gas biogénico en la mina de carbón La Ciscuda. Los resultados sugirieron que los metanógenos de este consorcio producían metano por vía hidrogenotrófica o de reducción de CO2.


ABSTRACT The work studied the methanogenic microbial consortium in a coal mine from the Bogotá basin in Colombia. Ex situ coal-enrichment cultures were established for in vitro growth and de novo gas production. Biogenic gas produced by cultures was analyzed by gas chromatography using thermal conductivity and flame ionization detectors. Cultures were used to isolate microbial specimens and to generate 16S rRNA gene libraries employing bacterial and archaeal primer sets. The gas chromatographic analysis showed methane production at 37 oC, but not at 60 oC, where CO2 was the major component of the biogenic gas. 16S rRNA gene sequence analysis of microbial isolates and clone libraries established that the methanogenic microbial consortium was formed by bacteria species from Bacillus and Gracilibacter genera plus archaea from the Methanothermobacter genus. This meth-anogenic microbial consortium was potentially responsible for biogenic gas generation in La Ciscuda coal mine. The results suggested that these methanogens produced methane by hydrogenotrophic or CO2 reduction pathways.

6.
Molecules ; 27(17)2022 Aug 27.
Artículo en Inglés | MEDLINE | ID: mdl-36080288

RESUMEN

Plants are sources of sunscreen ingredients that prevent cellular mutations involved in skin cancer and aging. This study investigated the sunscreen properties of the extracts from some ornamental plants growing in Colombia. The UV filter capability of the flower extracts obtained from Rosa centifolia L., Posoqueria latifolia (Rudge) Schult, and Ipomoea horsfalliae Hook. was examined. Photoprotection efficacies were evaluated using in vitro indices such as sun protection factor and critical wavelength. UVB antigenotoxicity estimates measured with the SOS Chromotest were also obtained. Extract cytotoxicity and genotoxicity were studied in human fibroblasts using the trypan blue exclusion and Comet assays, respectively. Major compounds of the promising flower extracts were identified by UHPLC-ESI+-Orbitrap-MS. The studied extracts showed high photoprotection efficacy and antigenotoxicity against UVB radiation, but only the P. latifolia extract showed broad-spectrum photoprotection at non-cytotoxic concentrations. The P. latifolia extract appeared to be safer for human fibroblast cells and the R. centifolia extract was shown to be moderately cytotoxic and genotoxic at the highest assayed concentrations. The I. horsfalliae extract was unequivocally cytotoxic and genotoxic. The major constituents of the promising extracts were as follows: chlorogenic acid, ecdysterone 20E, rhamnetin-rutinoside, cis-resveratrol-diglucoside, trans-resveratrol-diglucoside in P. latifolia; quercetin, quercetin-glucoside, quercetin-3-rhamnoside, kaempferol, kaempferol-3-glucoside, and kaempferol-rhamnoside in R. centifolia. The potential of the ornamental plants as sources of sunscreen ingredients was discussed.


Asunto(s)
Quempferoles , Protectores Solares , Flores , Glucósidos , Humanos , Extractos Vegetales/farmacología , Plantas , Quercetina , Protectores Solares/farmacología , Rayos Ultravioleta
7.
Photochem Photobiol ; 98(1): 254-261, 2022 01.
Artículo en Inglés | MEDLINE | ID: mdl-34403528

RESUMEN

Serratia marcescens is a bacterial species that produces an antibacterial pigment (Prodigiosin) showing a wide adaptive response to environmental stresses. The study aimed to investigate Prodigiosin production in S. marcescens wild-type strains, as well as its relation to photoprotection and antigenotoxicity against UVB. Prodigiosin yield was spectrophotometrically assayed in extracts of bacterial strains grown in different culture media. In vitro photoprotection efficacy was evaluated using the in vitro indices sun protection factor (SPFin vitro ) and critical wavelength (λc). The percentage of UVB antigenotoxicity estimates (%GI) in the SOS Chromotest was also evaluated. Correlation analysis was used to examine the relationship between Prodigiosin yield, SPFin vitro , %GI estimates and environmental traits (altitude, temperature, rainfall and solar irradiance). Prodigiosin yield in S. marcescens strains varied depending on culture media used for its growth, and it was correlated with environmental variables such as temperature and solar irradiance. SPFin vitro estimates were well correlated with Prodigiosin concentration and %GI values in the bacterial strains being studied. UVB photoprotective efficacy of the extracts obtained from S. marcescens strains depends on the strain's Prodigiosin yield and its antigenotoxic potential. The extracts with Prodigiosin yield higher than ~17 µg mL-1 could be used as sources of sunscreen ingredients.


Asunto(s)
Prodigiosina , Serratia marcescens , Colombia , Medios de Cultivo , Extractos Vegetales , Prodigiosina/farmacología , Serratia marcescens/fisiología
8.
Int J Radiat Biol ; 97(12): 1705-1715, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34586950

RESUMEN

INTRODUCTION: Plants can be sources of photoprotective/antigenotoxic compounds that prevent cellular mutations involved in skin cancer and aging by regulating UV-induced mutability. PURPOSE: The study was aimed at investigating the sunscreen properties of plants growing in Colombia. MATERIALS AND METHODS: Ultraviolet (UV) radiation-absorption capability of different plant extracts was examined. In vitro photoprotection efficacies were evaluated using in vitro indices such as sun protection factor (SPFin vitro) and critical wavelength (λc). Pearson correlation analysis was used to examine the relationship between SPFin vitro and complementary UVB- antigenotoxicity estimates (%GI) based on the SOS Chromotest database. The cytotoxicity in human fibroblasts was studied using the trypan blue exclusion assay. Major compounds of promising plant extracts were determined by gas chromatography coupled to mass spectrometry (GC/MS). RESULTS: We showed that plant extracts have sunscreen properties against UVB, whereas broad-spectrum radiation protection efficacy was poor. SPFin vitro and %GI were correlated (R = 0.71, p < .0001) for the plant extracts under study. Three extracts obtained from Achyrocline satureioides, Chromolaena pellia, and Lippia origanoides species resulted to possess high protection efficacy and relatively low cytotoxicity in human fibroblasts. These plant extracts contained major compounds such as α-pinene, trans-ß-caryophyllene, γ-muurolene, γ-cadinene and caryophyllene oxide in A. Satureioides extract, trans-ß-caryophyllene, caryophyllene oxide, squalene and α-amyrin in C. pellia extract, and p-cymene, carvacrol, trans-ß-caryophyllene and pinocembrin in L. origanoides extract. CONCLUSIONS: Plants growing in Colombia contain compounds that can be useful for potential sunscreens. SPFin vitro and %GI estimates were correlated, but %GI estimates were more sensitive to detecting activity at lower plant extract concentrations. Our results supported the need to use DNA damage detection assays as a complement to photoprotection efficacy measurement.


Asunto(s)
Lippia , Extractos Vegetales/farmacología , Protectores Solares , Colombia , Humanos , Rayos Ultravioleta/efectos adversos
9.
Asian J Androl ; 23(3): 306-313, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33269725

RESUMEN

Obesity is a major worldwide health problem that is related to most chronic diseases, including male infertility. Owing to its wide impact on health, mechanisms underlying obesity-related infertility remain unknown. In this study, we report that mice fed a high-fat diet (HFD) for over 2 months showed reduced fertility rates and increased germ cell apoptosis, seminiferous tubule degeneration, and decreased intratesticular estradiol (E2) and E2-to-testosterone ratio. Interestingly, we also detected a decrease in testicular fatty acid levels, behenic acid (C22:0), and docosahexaenoic acid (DHA, 22:6n-3), which may be related to the production of dysfunctional spermatozoa. Overall, we did not detect any changes in the frequency of seminiferous tubule stages, sperm count, or rate of in vitro capacitation. However, there was an increase in spontaneous and progesterone-induced acrosomal exocytosis (acrosome reaction) in spermatozoa from HFD-fed mice. These data suggest that a decrease in E2 and fatty acid levels influences spermatogenesis and some steps of acrosome biogenesis that will have consequences for fertilization. Thus, our results add new evidence about the adverse effect of obesity in male reproduction and suggest that the acrosomal reaction can also be affected under this condition.


Asunto(s)
Dieta Alta en Grasa/efectos adversos , Ácidos Docosahexaenoicos/deficiencia , Infertilidad Masculina/etiología , Testículo/metabolismo , Reacción Acrosómica/fisiología , Animales , Dieta Alta en Grasa/métodos , Modelos Animales de Enfermedad , Ácidos Docosahexaenoicos/análisis , Infertilidad Masculina/sangre , Masculino , Ratones , Obesidad/complicaciones , Espermatozoides/efectos de los fármacos
10.
Photochem Photobiol ; 95(4): 1010-1018, 2019 07.
Artículo en Inglés | MEDLINE | ID: mdl-30636010

RESUMEN

This work evaluated the photoprotective and antigenotoxic effects against ultraviolet B (UVB) radiation of flavonoid compounds apigenin, naringenin and pinocembrin. The photoprotective efficacy of these compounds was estimated using in vitro photoprotection indices, and the antigenotoxicity against UVB radiation was evaluated using the SOS chromotest and an enzymatic (proteinase K/T4 endonuclease V enzyme) comet assay in UV-treated Escherichia coli and human (HEK-293) cells, respectively. Naringenin and pinocembrin showed maximum UV-absorption peak in UVC and UVB zones, while apigenin showed UV-absorption capability from UVC to UVA range. These compounds acted as UV filters reducing UV-induced genotoxicity, both in bacteria and in human cells. The enzymatic comet assay resulted highly sensitive for detection of UVB-induced DNA damage in HEK-293 cells. In this work, the photoprotective potential of these flavonoids was widely discussed.


Asunto(s)
Apigenina/farmacología , Daño del ADN/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Flavanonas/farmacología , Escherichia coli/efectos de la radiación , Células HEK293 , Humanos , Protectores contra Radiación/farmacología , Rayos Ultravioleta
11.
FEMS Microbiol Lett ; 365(11)2018 06 01.
Artículo en Inglés | MEDLINE | ID: mdl-29718179

RESUMEN

Exposure to ultraviolet (UV) radiation blocks DNA replication and arrests cellular division in Escherichia coli. Restoration of chromosome replication involves nucleoid reorganization, which involves the participation of the recombination-catalyzing proteins RecA, RecO, RecR and RecN. In this work, we evaluated the influence of recN, uvrA and recJ gene mutations on post-irradiation nucleoid reorganization. We used isogenic E. coli strains that are defective for these genes to study post-irradiation kinetics of the nucleoid shape fractions using fluorescence microscopy. The results showed that in the wild-type strain, post-irradiation nucleoid reorganization occurs, which restores the nucleoid shape fractions in the cells to those observed prior to irradiation. First, the nucleoid condenses into the central area of the irradiated cell. Second, the nucleoid disperses along the cell. Third, the cell enters the chromosome replicative phase and cytokinesis. Escherichia coli cells with a recN mutation did not exhibit increased nucleoid condensation, but chromosome replication and cytokinesis occurred. In the uvrA and recJ strains, the condensation step was delayed compared to the wild-type strain, and chromosome replication and cytokinesis did not occur. The results are discussed with an emphasis on the functions of RecN, UvrA and RecJ in nucleoid reorganization in UV-irradiated E. coli cells.


Asunto(s)
Adenosina Trifosfatasas/metabolismo , Proteínas Bacterianas/metabolismo , Cromosomas Bacterianos/metabolismo , Enzimas de Restricción del ADN/metabolismo , Proteínas de Unión al ADN/metabolismo , Proteínas de Escherichia coli/metabolismo , Escherichia coli/efectos de la radiación , Exodesoxirribonucleasas/metabolismo , Mutación , Rayos Ultravioleta , Adenosina Trifosfatasas/genética , Proteínas Bacterianas/genética , Enzimas de Restricción del ADN/genética , ADN Bacteriano/metabolismo , Proteínas de Unión al ADN/genética , Escherichia coli/enzimología , Proteínas de Escherichia coli/genética , Exodesoxirribonucleasas/genética
12.
Photochem Photobiol ; 93(4): 1063-1072, 2017 07.
Artículo en Inglés | MEDLINE | ID: mdl-28178379

RESUMEN

The antigenotoxicity against ultraviolet radiation (UV)-induced DNA damage of essential oils (EO) from Lippia species was studied using SOS Chromotest. Based on the minimum concentration that significantly inhibits genotoxicity, the genoprotective potential of EO from highest to lowest was Lippia graveolens, thymol-RC ≈ Lippia origanoides, carvacrol-RC ≈ L. origanoides, thymol-RC > Lippia alba, citral-RC ≈ Lippia citriodora, citral-RC ≈ Lippia micromera, thymol-RC > L. alba, myrcenone-RC. EO from L. alba, carvone/limonene-RC, L. origanoides, α-phellandrene-RC and L. dulcis, trans-ß-caryophyllene-RC did not reduce the UV genotoxicity at any of the doses tested. A gas chromatography with flame ionization detection analysis (GC-FID) was conducted to evaluate the solubility of the major EO constituents under our experimental conditions. GC-FID analysis showed that, at least partially, major EO constituents were water-soluble and therefore, they were related with the antigenotoxicity detected for EO. Constituents such as p-cymene, geraniol, carvacrol, thymol, citral and 1,8-cineole showed antigenotoxicity. The antioxidant activity of EO constituents was also determined using the oxygen radical antioxidant capacity (ORAC) assay. The results showed that the antigenotoxicity of the EO constituents was unconnected with their antioxidant activity. The antigenotoxicity to different constituent binary mixtures suggests that synergistic effects can occur in some of the studied EO.


Asunto(s)
Antimutagênicos/farmacología , Daño del ADN , Lippia/química , Aceites Volátiles/farmacología , Aceites de Plantas/farmacología , Rayos Ultravioleta , Antioxidantes/farmacología , Cromatografía de Gases , Lippia/clasificación , Respuesta SOS en Genética , Especificidad de la Especie
13.
Int J Radiat Biol ; 92(6): 321-8, 2016 06.
Artículo en Inglés | MEDLINE | ID: mdl-26967458

RESUMEN

Purpose In this paper, the contribution of different genes involved in DNA repair for both survival and SOS induction in Escherichia coli mutants exposed to ultraviolet B radiation (UVB, [wavelength range 280-315 nm]) was evaluated. Materials and methods E. coli strains defective in uvrA, oxyR, recO, recN, recJ, exoX, recB, recD or xonA genes were used to determine cell survival. All strains also had the genetic sulA::lacZ fusion, which allowed for the quantification of SOS induction through the SOS Chromotest. Results Five gene products were particularly important for survival, as follows: UvrA > RecB > RecO > RecJ > XonA. Strains defective in uvrA and recJ genes showed elevated SOS induction compared with the wild type, which remained stable for up to 240 min after UVB-irradiation. In addition, E. coli strains carrying the recO or recN mutation showed no SOS induction. Conclusions The nucleotide excision and DNA recombination pathways were equally used to repair UVB-induced DNA damage in E. coli cells. The sulA gene was not turned off in strains defective in UvrA and RecJ. RecO protein was essential for processing DNA damage prior to SOS induction. In this study, the roles of DNA repair proteins and their contributions to the mechanisms that induce SOS genes in E. coli are proposed.


Asunto(s)
Supervivencia Celular/efectos de la radiación , Escherichia coli/fisiología , Escherichia coli/efectos de la radiación , Respuesta SOS en Genética/fisiología , Respuesta SOS en Genética/efectos de la radiación , Rayos Ultravioleta , Proteínas Bacterianas/metabolismo , Supervivencia Celular/fisiología , Relación Dosis-Respuesta en la Radiación , Escherichia coli/citología , Dosis de Radiación
14.
Genet Mol Biol ; 35(2): 503-14, 2012 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-22888301

RESUMEN

In this work, the toxicity and genotoxicity of organic solvents (acetone, carbon tetrachloride, dichloromethane, dimethylsulfoxide, ethanol, ether and methanol) were studied using the SOS chromotest. The influence of these solvents on the direct genotoxicity induced by the mutagens mitomycin C (MMC) and 4-nitroquinoline-1-oxide (4-NQO) were also investigated. None of the solvents were genotoxic in Escherichia coli PQ37. However, based on the inhibition of protein synthesis assessed by constitutive alkaline phosphatase activity, some solvents (carbon tetrachloride, dimethylsulfoxide, ethanol and ether) were toxic and incompatible with the SOS chromotest. Solvents that were neither toxic nor genotoxic to E. coli (acetone, dichloromethane and methanol) significantly reduced the genotoxicity of MMC and 4-NQO. When these solvents were used to dissolve vitamin E they increased the antigenotoxic activity of this compound, possibly through additive or synergistic effects. The relevance of these results is discussed in relation to antigenotoxic studies. These data indicate the need for careful selection of an appropriate diluent for the SOS chromotest since some solvents can modulate genotoxicity and antigenotoxicity.

15.
Genet Mol Biol ; 34(3): 479-88, 2011 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-21931523

RESUMEN

The present work evaluated the chemical composition and the DNA protective effect of the essential oils (EOs) from Lippia alba against bleomycin-induced genotoxicity. EO constituents were determined by Gas Chromatography/Mass Spectrometric (GC-MS) analysis. The major compounds encountered being citral (33% geranial and 25% neral), geraniol (7%) and trans-ß-caryophyllene (7%) for L. alba specimen COL512077, and carvone (38%), limonene (33%) and bicyclosesquiphellandrene (8%) for the other, COL512078. The genotoxicity and antigenotoxicity of EO and the compounds citral, carvone and limonene, were assayed using the SOS Chromotest in Escherichia coli. The EOs were not genotoxic in the SOS chromotest, but one of the major compound (limonene) showed genotoxicity at doses between 97 and 1549 mM. Both EOs protected bacterial cells against bleomycin-induced genotoxicity. Antigenotoxicity in the two L. alba chemotypes was related to the major compounds, citral and carvone, respectively. The results were discussed in relation to the chemopreventive potential of L. alba EOs and its major compounds.

16.
Genet Mol Biol ; 34(4): 681-8, 2011 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-22215974

RESUMEN

The aim of this study was to use the Comet assay to assess genetic damage in the direct-developing frog Eleutherodactylus johnstonei. A DNA diffusion assay was used to evaluate the effectiveness of alkaline, enzymatic and alkaline/enzymatic treatments for lysing E. johnstonei blood cells and to determine the amount of DNA strand breakage associated with apoptosis and necrosis. Cell sensitivity to the mutagens bleomycin (BLM) and 4-nitro-quinoline-1-oxide (4NQO) was also assessed using the Comet assay, as was the assay reproducibility. Alkaline treatment did not lyse the cytoplasmic and nuclear membranes of E. johnstonei blood cells, whereas enzymatic digestion with proteinase K (40 µg/mL) yielded naked nuclei. The contribution of apoptosis and necrosis (assessed by the DNA diffusion assay) to DNA damage was estimated to range from 0% to 8%. BLM and 4NQO induced DNA damage in E. johnstonei blood cells at different concentrations and exposure times. Dose-effect curves with both mutagens were highly reproducible and showed consistently low coefficients of variation (CV ≤ 10%). The results are discussed with regard to the potential use of the modified Comet assay for assessing the exposure of E. johnstonei to herbicides in ecotoxicological studies.

17.
Genet. mol. biol ; 34(3): 479-488, 2011. ilus
Artículo en Inglés | LILACS | ID: lil-595983

RESUMEN

The present work evaluated the chemical composition and the DNA protective effect of the essential oils (EOs) from Lippia alba against bleomycin-induced genotoxicity. EO constituents were determined by Gas Chromatography/Mass Spectrometric (GC-MS) analysis. The major compounds encountered being citral (33 percent geranial and 25 percent neral), geraniol (7 percent) and trans-β-caryophyllene (7 percent) for L. alba specimen COL512077, and carvone (38 percent), limonene (33 percent) and bicyclosesquiphellandrene (8 percent) for the other, COL512078. The genotoxicity and antigenotoxicity of EO and the compounds citral, carvone and limonene, were assayed using the SOS Chromotest in Escherichia coli. The EOs were not genotoxic in the SOS chromotest, but one of the major compound (limonene) showed genotoxicity at doses between 97 and 1549 mM. Both EOs protected bacterial cells against bleomycin-induced genotoxicity. Antigenotoxicity in the two L. alba chemotypes was related to the major compounds, citral and carvone, respectively. The results were discussed in relation to the chemopreventive potential of L. alba EOs and its major compounds.


Asunto(s)
Genotoxicidad , Lippia/química , Aceites Volátiles , Bleomicina , Lippia/toxicidad
18.
Genet. mol. biol ; 34(4): 681-688, 2011. ilus, tab
Artículo en Inglés | LILACS | ID: lil-605946

RESUMEN

The aim of this study was to use the Comet assay to assess genetic damage in the direct-developing frog Eleutherodactylus johnstonei. A DNA diffusion assay was used to evaluate the effectiveness of alkaline, enzymatic and alkaline/enzymatic treatments for lysing E. johnstonei blood cells and to determine the amount of DNA strand breakage associated with apoptosis and necrosis. Cell sensitivity to the mutagens bleomycin (BLM) and 4-nitroquinoline-1-oxide (4NQO) was also assessed using the Comet assay, as was the assay reproducibility. Alkaline treatment did not lyse the cytoplasmic and nuclear membranes of E. johnstonei blood cells, whereas enzymatic digestion with proteinase K (40 !g/mL) yielded naked nuclei. The contribution of apoptosis and necrosis (assessed by the DNA diffusion assay) to DNA damage was estimated to range from 0 percent to 8 percent. BLM and 4NQO induced DNA damage in E. johnstonei blood cells at different concentrations and exposure times. Dose-effect curves with both mutagens were highly reproducible and showed consistently low coefficients of variation (CV < 10 percent). The results are discussed with regard to the potential use of the modified Comet assay for assessing the exposure of E. johnstonei to herbicides in ecotoxicological studies.


Asunto(s)
Bleomicina , Ensayo Cometa
19.
Environ Toxicol ; 25(2): 130-6, 2010 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-19399845

RESUMEN

Amifostine is the most effective radioprotector known and the only one accepted for clinical use in cancer radiotherapy. In this work, the antigenotoxic effect of amifostine against gamma-rays was studied in Escherichia coli cells deficient in DNA damage repair activities. Assays of irradiated cells treated with amifostine showed that the drug reduced the genotoxicity induced by radiation in E. coli wild-type genotypes and in uvr, recF, recB, recB-recC-recF mutant strains, but not in recN defective cells. Thus, the mechanism of DNA protection by amifostine against gamma-radiation-induced genotoxicity appears to involve participation of the RecN protein that facilitates repair of DNA double-strand breaks. The results are discussed in relation to amifostine's chemopreventive potential.


Asunto(s)
Amifostina/farmacología , Daño del ADN/efectos de los fármacos , Proteínas de Escherichia coli/metabolismo , Escherichia coli/efectos de los fármacos , Rayos gamma , Protectores contra Radiación/farmacología , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Daño del ADN/efectos de la radiación , Reparación del ADN/efectos de los fármacos , Reparación del ADN/efectos de la radiación , Enzimas de Restricción del ADN/genética , Enzimas de Restricción del ADN/metabolismo , Escherichia coli/genética , Escherichia coli/efectos de la radiación , Proteínas de Escherichia coli/genética
20.
Fitoterapia ; 81(5): 343-9, 2010 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-19874875

RESUMEN

The present work evaluated the chemical composition of the essential oils (EO) obtained from Lippia origanoides and their DNA protective effect against bleomycin-induced genotoxicity. L. origanoides EO chemical composition was determined by gas chromatography-mass spectrometry (GC-MS). The major compounds of the L. origanoides EOs were thymol (34-58%) and carvacrol (26%). The antigenotoxic effects of the EOs, major compounds and standard compound (epigallocatechin gallate) were assayed in co-incubation procedures using the SOS chromotest in Escherichia coli. Both EOs and their major compounds protected bacterial cells against bleomycin-induced genotoxicity indicating that these two compounds were principally responsible for the antigenotoxicity detected in the oils. Thymol and carvacrol antigenotoxicity was lower than those observed with epigallocatechin gallate. The results were discussed in relation to the chemopreventive potential of L. origanoides EOs and their major components, carvacrol and thymol.


Asunto(s)
Antimutagênicos/farmacología , Daño del ADN/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Lippia/química , Monoterpenos/farmacología , Aceites Volátiles/farmacología , Extractos Vegetales/farmacología , Antimutagênicos/análisis , Bleomicina , Catequina/análogos & derivados , Técnicas de Cultivo de Célula , Cimenos , Escherichia coli/genética , Flores , Cromatografía de Gases y Espectrometría de Masas , Monoterpenos/análisis , Aceites Volátiles/química , Extractos Vegetales/química , Hojas de la Planta , Timol/análisis , Timol/farmacología
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