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1.
Molecules ; 26(11)2021 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-34198881

RESUMO

Euterpe oleracea Mart. (Arecaceae) is an endogenous palm tree from the Amazon region. Its seeds correspond to 85% of the fruit's weight, a primary solid residue generated from pulp production, the accumulation of which represents a potential source of pollution and environmental problems. As such, this work aimed to quantify and determine the phytochemical composition of E. oleracea Mart. seeds from purple, white, and BRS-Pará açaí varieties using established analytical methods and also to evaluate it as an eco-friendly corrosion inhibitor. The proanthocyanidin quantification (n-butanol/hydrochloric acid assay) between varieties was 6.4-22.4 (w/w)/dry matter. Extract characterization showed that all varieties are composed of B-type procyanidin with a high mean degree of polymerization (mDP ≥ 10) by different analytical methodologies to ensure the results. The purple açaí extract, which presented 22.4% (w/w) proanthocyanidins/dry matter, was tested against corrosion of carbon steel AISI 1020 in neutral pH. The crude extract (1.0 g/L) was effective in controlling corrosion on the metal surface for 24 h. Our results demonstrated that the extracts rich in polymeric procyanidins obtained from industrial açaí waste could be used to inhibit carbon steel AISI 1020 in neutral pH as an abundant, inexpensive, and green source of corrosion inhibitor.


Assuntos
Euterpe/química , Proantocianidinas/farmacologia , Aço/química , Corrosão , Química Verde , Extratos Vegetais/química , Proantocianidinas/química , Sementes/química
2.
J Nat Prod ; 82(3): 566-572, 2019 03 22.
Artigo em Inglês | MEDLINE | ID: mdl-30817148

RESUMO

Lippia species share various pharmacological activities and are used in traditional cooking and medicine worldwide. Combined chromatographic techniques such as column chromatography, high-performance liquid chromatography, and countercurrent chromatography led to the purification of two new antifungal phenylpropanoid glycosides, lippiarubelloside A (1) and lippiarubelloside B (2), by bioactivity-directed fractionation of an ethanol-soluble extract from Lippia rubella, in addition to the known active related compounds forsythoside A (3), verbascoside (4), isoverbascoside (5), and poliumoside (6). The structures of compounds 1 and 2 were determined by comparison of their NMR spectroscopic data with the prototype active compound 4. Cryptococcus neoformans, which causes opportunistic lung infections, was sensitive to compounds 1-6 in the concentration range of 15-125 µg/mL. A synergistic effect (FICindex = 0.5) between 3 and amphotericin B was demonstrated. The glycosylated flavonoids pectolinarin (7), linarin (8), and siparunoside (9) were also isolated.


Assuntos
Antifúngicos/farmacologia , Glicosídeos/farmacologia , Lippia/química , Fenilpropionatos/farmacologia , Antifúngicos/química , Candida/efeitos dos fármacos , Cryptococcus/efeitos dos fármacos , Glicosídeos/química , Fenilpropionatos/química , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Análise Espectral/métodos
3.
Mem Inst Oswaldo Cruz ; 113(8): e180120, 2018 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-29947713

RESUMO

BACKGROUND: Melanin production has been associated with virulence in various pathogenic fungi, including Fonsecaea pedrosoi, the major etiological agent for chromoblastomycosis, a subcutaneous fungal disease that occurs in South America. OBJECTIVE: The aim of this study was to evaluate the effects of acid-basic extracted F. pedrosoi melanin particles and fungal cell ghosts obtained by Novozym 234 treatment on their ability to activate the human complement system. METHODS: The ability of melanin particles and fungal cell ghosts to activate the human complement system was evaluated by complement consumption, immunofluorescence, and enzyme-linked immunosorbent assay (ELISA). FINDINGS: Unsensitised melanin particles and melanin ghosts presented complement consumption of 82.67 ± 2.08% and 96.04 ± 1.13%, respectively. Immunofluorescence assays revealed intense deposition of the C3 and C4 fragments on the surface of melanin particles and ghosts extracted from F. pedrosoi. Deposition of the C3, C4, and C5 fragments onto melanin samples and zymosan was confirmed by ELISA. Deposition of small amounts of C1q and C9 onto melanin samples and zymosan was detected by ELISA. CONCLUSION: Fonsecaea pedrosoi melanin particles and fungal cell ghosts activated the complement system mainly through an alternative pathway.


Assuntos
Ascomicetos/química , Ativação do Complemento , Proteínas do Sistema Complemento/imunologia , Melaninas/farmacologia , Animais , Ensaio de Imunoadsorção Enzimática , Imunofluorescência , Humanos , Melaninas/biossíntese , Melaninas/isolamento & purificação
4.
Molecules ; 23(6)2018 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-29865153

RESUMO

Monohexosylceramides (CMHs) are highly conserved fungal glycosphingolipids playing a role in several cellular processes such as growth, differentiation and morphological transition. In this study, we report the isolation, purification and chemical characterization of CMHs from Rhizopus stolonifer and R. microspores. Using positive ion mode ESI-MS, two major ion species were observed at m/z 750 and m/z 766, respectively. Both ion species consisted of a glucose/galactose residue attached to a ceramide moiety containing 9-methyl-4,8-sphingadienine with an amidic linkage to a hydroxylated C16:0 fatty acid. The antimicrobial activity of CMH was evaluated against Gram positive and Gram negative bacteria using the agar diffusion assay. CMH from both Rhizopus species inhibited the growth of Bacillus terrae, Micrococcus luteus (M. luteus) and Pseudomonas stutzeri (P. stutzeri) with a MIC50 of 6.25, 6.25 and 3.13 mg/mL, respectively. The bactericidal effect was detected only for M. luteus and P. stutzeri, with MBC values of 25 and 6.25 mg/mL, respectively. Furthermore, the action of CMH on the biofilm produced by methicillin-resistant Staphylococcus aureus (MRSA) was analyzed using 12.5 and 25 mg/mL of CMH from R. microsporus. Total biofilm biomass, biofilm matrix and viability of the cells that form the biofilm structure were evaluated. CMH from R. microsporus was able to inhibit the MRSA biofilm formation in all parameters tested.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Cerebrosídeos/isolamento & purificação , Cerebrosídeos/farmacologia , Rhizopus/química , Antibacterianos/química , Biomassa , Brasil , Cerebrosídeos/química , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Espectrometria de Massas por Ionização por Electrospray
5.
Molecules ; 22(4)2017 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-28422054

RESUMO

Strategies for the control of sulfate-reducing bacteria (SRB) in the oil industry involve the use of high concentrations of biocides, but these may induce bacterial resistance and/or be harmful to public health and the environment. Essential oils (EO) produced by plants inhibit the growth of different microorganisms and are a possible alternative for controlling SRB. We aimed to characterize the bacterial community of produced water obtained from a Brazilian petroleum facility using molecular methods, as well as to evaluate the antimicrobial activity of EO from different plants and their major components against Desulfovibrio alaskensis NCIMB 13491 and against SRB growth directly in the produced water. Denaturing gradient gel electrophoresis revealed the presence of the genera Pelobacter and Marinobacterium, Geotoga petraea, and the SRB Desulfoplanes formicivorans in our produced water samples. Sequencing of dsrA insert-containing clones confirmed the presence of sequences related to D. formicivorans. EO obtained from Citrus aurantifolia, Lippia alba LA44 and Cymbopogon citratus, as well as citral, linalool, eugenol and geraniol, greatly inhibited (minimum inhibitory concentration (MIC) = 78 µg/mL) the growth of D. alaskensis in a liquid medium. The same MIC was obtained directly in the produced water with EO from L. alba LA44 (containing 82% citral) and with pure citral. These findings may help to control detrimental bacteria in the oil industry.


Assuntos
Antibacterianos/farmacologia , Bactérias/efeitos dos fármacos , Bactérias/metabolismo , Indústria de Petróleo e Gás , Óleos Voláteis/farmacologia , Sulfatos/metabolismo , Água , Bactérias/genética , Testes de Sensibilidade Microbiana
6.
Int J Med Microbiol ; 305(8): 937-47, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26558847

RESUMO

The capacity to form biofilm is considered a protective mechanism that allows the bacteria to survive and proliferate in hostile environments, facilitating the maintenance of the infectious process. Recently, biofilm has become a topic of interest in the study of the human pathogen group A Streptococcus (GAS). Although GAS has not been associated with infection on medical implants, the presence of microcolonies embedded in an extracellular matrix on infected tissues has been reported. Despite the similarity between GAS and Streptococcus dysgalactiae subspecies equisimilis (SDSE), there are no studies in the literature describing the production of biofilm by SDSE. In this work, we assessed and characterized biofilm development among SDSE human isolates of group C. The in vitro data showed that 59.3% of the 118 isolates tested were able to form acid-induced biofilm on glass, and 28% formed it on polystyrene surfaces. More importantly, biofilm was also formed in a foreign body model in mice. The biofilm structure was analyzed by confocal laser scanning microscopy, transmission electron microscopy, and scanning electron microscopy. Long fibrillar-like structures were observed by scanning electron microscopy. Additionally, the expression of a pilus associated gene of SDSE was increased for in vitro sessile cells compared with planktonics, and when sessile cells were collected from biofilms formed in the animal model compared with that of in vitro model. Results obtained from the immunofluorescence microscopy indicated the biofilm was immunogenic. Our data also suggested a role for proteins, exopolysaccharide and extracellular DNA in the formation and accumulation of biofilm by SDSE.


Assuntos
Biofilmes/crescimento & desenvolvimento , Infecções Estreptocócicas/microbiologia , Streptococcus/isolamento & purificação , Streptococcus/fisiologia , Animais , Proteínas de Bactérias/análise , DNA Bacteriano/análise , Modelos Animais de Doenças , Corpos Estranhos/complicações , Humanos , Masculino , Camundongos Endogâmicos BALB C , Microscopia Confocal , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência , Polissacarídeos Bacterianos/análise , Streptococcus/crescimento & desenvolvimento
7.
Mycopathologia ; 179(3-4): 231-42, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25502596

RESUMO

Phialophora verrucosa is one of the etiologic agents of chromoblastomycosis, a fungal infection that affects cutaneous and subcutaneous tissues. This disease is chronic, recurrent and difficult to treat. Several studies have shown that secreted peptidases by fungi are associated with important pathophysiological processes. Herein, we have identified and partially characterized the peptidase activity secreted by P. verrucosa conidial cells. Using human serum albumin as substrate, the best hydrolysis profile was detected at extreme acidic pH (3.0) and at 37 °C. The enzymatic activity was completely blocked by classical metallopeptidase inhibitors/chelating agents as 1,10-phenanthroline and EGTA. Zinc ions stimulated the metallo-type peptidase activity in a dose-dependent manner. Several proteinaceous substrates were cleaved, in different extension, by the P. verrucosa metallopeptidase activity, including immunoglobulin G, fibrinogen, collagen types I and IV, fibronectin, laminin and keratin; however, mucin and hemoglobin were not susceptible to proteolysis. As metallopeptidases participate in different cellular metabolic pathways in fungal cells, we also tested the influence of 1,10-phenanthroline and EGTA on P. verrucosa development. Contrarily to EGTA, 1,10-phenanthroline inhibited the fungal viability (MIC 0.8 µg/ml), showing fungistatic effect, and induced profound morphological alterations as visualized by transmission electron microscopy. In addition, 1,10-phenanthroline arrested the filamentation process in P. verrucosa. Our results corroborate the supposition that metallopeptidase inhibitors/chelating agents have potential to control crucial biological events in fungal agents of chromoblastomycosis.


Assuntos
Antifúngicos/farmacologia , Proteínas Fúngicas/metabolismo , Metaloproteases/metabolismo , Fenantrolinas/farmacologia , Phialophora/efeitos dos fármacos , Phialophora/enzimologia , Esporos Fúngicos/crescimento & desenvolvimento , Humanos , Micoses/microbiologia , Phialophora/crescimento & desenvolvimento , Sistemas de Translocação de Proteínas/metabolismo , Transporte Proteico , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/enzimologia
8.
Cancer Invest ; 32(6): 226-35, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24745610

RESUMO

Current therapies for glioblastoma multiforme (GBM) are not effective. This study investigated the activity of the M. officinalis essential oil (EO) and its major component (citral) in GBM cell lines. Both EO and citral decreased the viability and induced apoptosis of GBM cells as demonstrated by DNA fragmentation and caspase-3 activation. Antioxidant prevented citral-induced death, indicating its dependence on the production of reactive oxygen species. Citral downmodulated the activity and inhibited the expression of multidrug resistance associated protein 1 (MRP1). These results show that EO, through its major component, citral, may be of potential interest for the treatment of GBM.


Assuntos
Apoptose/efeitos dos fármacos , Melissa/química , Monoterpenos/farmacologia , Óleos Voláteis/farmacologia , Monoterpenos Acíclicos , Caspase 3/biossíntese , Linhagem Celular Tumoral , Fragmentação do DNA/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Glioblastoma/tratamento farmacológico , Humanos , Monoterpenos/química , Proteínas Associadas à Resistência a Múltiplos Medicamentos/biossíntese , Óleos Voláteis/química , Espécies Reativas de Oxigênio/metabolismo
9.
J Enzyme Inhib Med Chem ; 29(1): 12-7, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23323991

RESUMO

Abstract Lippia alba (Miller) N.E. Brown is an aromatic plant known locally as "Erva-cidreira-do-campo" that has great importance in Brazilian folk medicine. The aim of our study was to evaluate the antidermatophytic potential of linalool-rich essential oil (EO) from L. alba and analyze the ability of this EO to inhibit peptidase and keratinase activities, which are important virulence factors in dermatophytes. The minimum inhibitory concentrations (MICs) of L. alba EO were 39, 156 and 312 µg/mL against Trichophyton rubrum, Epidermophyton floccosum and Microsporum gypseum, respectively. To evaluate the influence of L. alba EO on the proteolytic and keratinolytic activities of these dermatophytes, specific inhibitory assays were performed. The results indicated that linalool-rich EO from L. alba inhibited the activity of proteases and keratinases secreted from dermatophytes, and this inhibition could be a possible mechanism of action against dermatophytes. Due to the effective antidermatophytic activity of L. alba EO, further experiments should be performed to explore the potential of this linalool-rich EO as an alternative antifungal therapy.


Assuntos
Arthrodermataceae/enzimologia , Lippia/química , Monoterpenos/análise , Óleos Voláteis/farmacologia , Peptídeo Hidrolases/efeitos dos fármacos , Inibidores de Proteases/farmacologia , Monoterpenos Acíclicos , Eletroforese em Gel de Poliacrilamida , Concentração de Íons de Hidrogênio , Testes de Sensibilidade Microbiana , Óleos Voláteis/química
10.
BMC Microbiol ; 13: 29, 2013 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-23387945

RESUMO

BACKGROUND: Lippia sidoides Cham., also known as pepper-rosmarin, produces an essential oil in its leaves that is currently used by the pharmaceutical, perfumery and cosmetic industries for its antimicrobial and aromatic properties. Because of the antimicrobial compounds (mainly thymol and carvacrol) found in the essential oil, we believe that the endophytic microorganisms found in L. sidoides are selected to live in different parts of the plant. RESULTS: In this study, the endophytic microbial communities from the stems and leaves of four L. sidoides genotypes were determined using cultivation-dependent and cultivation-independent approaches. In total, 145 endophytic bacterial strains were isolated and further grouped using either ERIC-PCR or BOX-PCR, resulting in 76 groups composed of different genera predominantly belonging to the Gammaproteobacteria. The endophytic microbial diversity was also analyzed by PCR-DGGE using 16S rRNA-based universal and group-specific primers for total bacteria, Alphaproteobacteria, Betaproteobacteria and Actinobacteria and 18S rRNA-based primers for fungi. PCR-DGGE profile analysis and principal component analysis showed that the total bacteria, Alphaproteobacteria, Betaproteobacteria and fungi were influenced not only by the location within the plant (leaf vs. stem) but also by the presence of the main components of the L. sidoides essential oil (thymol and/or carvacrol) in the leaves. However, the same could not be observed within the Actinobacteria. CONCLUSION: The data presented here are the first step to begin shedding light on the impact of the essential oil in the endophytic microorganisms in pepper-rosmarin.


Assuntos
Antibacterianos/farmacologia , Bactérias/classificação , Biota , Endófitos/classificação , Lippia/química , Lippia/microbiologia , Óleos Voláteis/farmacologia , Antibacterianos/análise , Bactérias/efeitos dos fármacos , Bactérias/genética , Bactérias/isolamento & purificação , Análise por Conglomerados , Impressões Digitais de DNA , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Endófitos/efeitos dos fármacos , Endófitos/genética , Endófitos/isolamento & purificação , Dados de Sequência Molecular , Óleos Voláteis/análise , Filogenia , Folhas de Planta/química , Folhas de Planta/microbiologia , Caules de Planta/química , Caules de Planta/microbiologia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
11.
BMC Complement Altern Med ; 13: 107, 2013 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-23680079

RESUMO

BACKGROUND: Teas from the husk fiber of Cocos nucifera are used in the folk medicine to treat arthritis and other inflammatory processes. Some works show that some varieties have biological activities. However, one of the main variety of the species, C. nucifera var. typica, known in Brazil as "gigante", was not studied yet. Thus, this study evaluates if this variety has the anti-inflammatory and antimicrobial activities already reported in other varieties. METHODS: C. nucifera aqueous crude extract (10, 50, and 100 mg/kg) and the reference drugs morphine (1 mg/kg) and acetylsalicylic acid (100 mg/kg) were evaluated in models of inflammation (formalin-induced licking and subcutaneous air pouch). The antioxidant activity was evaluated by 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) photometric assay and compared with those of the standards (quercetin, rutin, and ascorbic acid). The extract was also screened against Candida albicans, Escherichia coli, Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus (MRSA), in the agar diffusion method. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were determined by the broth micro-dilution assay. Activities of combinations of the extract and antibiotics (methicillin or vancomycin) against MRSA were evaluated using checkerboard assays. RESULTS: The extract significantly inhibited the time that the animals spent licking the formalin-injected paws (second phase). The extract also inhibited the inflammatory process induced by subcutaneous carrageenan injection by reducing cell migration, protein extravasation, and TNF-α production. Additionally, the extract showed an antioxidant potential in vitro as good as standards in their antioxidant activity. The extract was active only against S. aureus and MRSA. MIC and the bactericidal concentrations were identical (1,024 µg/ml). The extract and methicillin acted synergistically against the clinical MRSA isolate, whereas an indifferent effect was detected when the extract was combined with vancomycin. CONCLUSIONS: The extract exhibits anti-inflammatory activity through the inhibition of the cell migration. The mixture of extract constituents and methicillin could lead to the development of a new combination antibiotic against MRSA infections.


Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Cocos/química , Extratos Vegetais/farmacologia , Animais , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Anti-Inflamatórios/química , Antioxidantes/química , Bactérias/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Humanos , Inflamação/tratamento farmacológico , Inflamação/fisiopatologia , Masculino , Camundongos , Extratos Vegetais/química
12.
Mycopathologia ; 176(3-4): 183-9, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23943428

RESUMO

The possible role of sialic acids in host cells-fungi interaction and their association with glycoproteins were evaluated using a clinical isolate of the dimorphic fungus Mucor polymorphosporus. Lectin-binding assays with spores and yeast cells denoted the presence of surface sialoglycoconjugates containing 2,3- and 2,6-linked sialylglycosyl groups. Western blotting with peroxidase-labeled Limulus polyphemus agglutinin revealed the occurrence of different sialoglycoprotein types in both cell lysates and cell wall protein extracts of mycelia, spores, and yeasts of M. polymorphosporus. Sialic acids contributed to the surface negative charge of spores and yeast forms as evaluated by adherence to a cationic substrate. Sialidase-treated spores were less resistant to phagocytosis by human neutrophils and monocytes from healthy individuals than control (untreated) fungal suspensions. The results suggest that sialic acids are terminal units of various glycoproteins of M. polymorphosporus, contributing to negative charge of yeasts and spore cells and protecting infectious propagules from destruction by host cells.


Assuntos
Sangue/imunologia , Mucor/química , Mucor/imunologia , Fagócitos/imunologia , Fagocitose/efeitos dos fármacos , Sialoglicoproteínas/imunologia , Sialoglicoproteínas/metabolismo , Humanos , Hifas/química , Hifas/imunologia , Lectinas/metabolismo , Mucor/isolamento & purificação , Mucormicose/microbiologia , Ligação Proteica , Esporos Fúngicos/química , Esporos Fúngicos/imunologia , Eletricidade Estática
13.
Molecules ; 17(6): 6305-16, 2012 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-22634841

RESUMO

The antimicrobial activities of the isomers and enantiomers of pinene were evaluated against bacterial and fungal cells. The agar diffusion test showed that only the positive enantiomers of the α- and ß-isomers of pinene were active. The minimal inhibitory concentration (MIC) and minimal microbicidal concentration (MMC) of these monoterpenes were also determined, confirming that the positive enantiomers exhibited microbicidal activity against all fungi and bacteria tested with MICs ranging from 117 to 4,150 µg/mL. However, no antimicrobial activity was detected with the negative enantiomers up to 20 mg/mL. Time-kill curves showed that (+)-α-pinene and (+)-ß-pinene were highly toxic to Candida albicans, killing 100% of inoculum within 60 min. By contrast, the bactericidal effect occurred after 6 h in methicillin-resistant Staphylococcus aureus (MRSA). In combination with commercial antimicrobials, ciprofloxacin plus (+)-α-pinene or (+)-ß-pinene presented synergistic activity against MRSA whereas an indifferent effect against all fungi was detected when amphotericin B was combined with the positive enantiomers of pinene. The potential of (+)-α-pinene and (+)-ß-pinene to inhibit phospholipase and esterase activities was also evaluated, and the best inhibition results were obtained with Cryptococcus neoformans. C. albicans biofilm formation was prevented with the MIC concentration of (+)-α-pinene and twice the MIC value of (+)-ß-pinene. Finally, the cytotoxicity of the positive enantiomers of pinene to murine macrophages was evaluated, and 250 µg/mL of (+)-α-pinene and (+)-ß-pinene reduced the cell viability to 66.8% and 57.7%, respectively.


Assuntos
Anti-Infecciosos/farmacologia , Compostos Bicíclicos com Pontes/farmacologia , Monoterpenos/farmacologia , Animais , Antibacterianos/farmacologia , Anti-Infecciosos/química , Antifúngicos/farmacologia , Bactérias/efeitos dos fármacos , Bactérias/metabolismo , Monoterpenos Bicíclicos , Biofilmes/efeitos dos fármacos , Compostos Bicíclicos com Pontes/química , Sinergismo Farmacológico , Esterases/antagonistas & inibidores , Fungos/efeitos dos fármacos , Fungos/metabolismo , Macrófagos/efeitos dos fármacos , Camundongos , Testes de Sensibilidade Microbiana , Monoterpenos/química , Fosfolipases/antagonistas & inibidores
14.
World J Microbiol Biotechnol ; 28(3): 953-62, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22805816

RESUMO

In a search for an antifungal substance with activity against the dermatophyte fungus Trichophyton rubrum, strain POC 115 was chosen among different Paenibacillus strains for its phenotypic and genetic characterization and for preliminary characterization of its antimicrobial substance. Strain POC 115 was identified as belonging to Paenibacillus kribbensis. Physico-chemical characterization of the antimicrobial substance showed that it was not stable during heat and organic solvents treatments, but its activity was preserved at a wide range of pH and after treatment with pronase E, trypsin and DNase I. The crude concentrated supernatant of POC 115 culture was partially purified and the fraction presenting antimicrobial activity was further analyzed by UPLC/Mass Spectrometry. Two peaks were observed at 2.02 (mass 1,207 D) and 2.71 (mass 1,014 D) min in the mass chromatogram. The antimicrobial substance produced by POC 115 was correlated to iturin family compounds based on a set of primers designed for the amplification of PKS operon in the POC 115 genome. As happens with the mode of action of the antibiotics of the iturin group, the AMS produced by POC 115 caused the disruption of cytoplasmic membrane of T. rubrum and the subsequent withdraw of the intracellular material. This is the first report on the production of antimicrobial substances in P. kribbensis, and it may be of great relevance as an alternative or supplementary substance to antifungal drugs currently used against dermatophytes.


Assuntos
Antibiose , Antifúngicos/farmacologia , Paenibacillus/fisiologia , Trichophyton/crescimento & desenvolvimento , Antifúngicos/química , Antifúngicos/isolamento & purificação , Antifúngicos/metabolismo , Técnicas de Tipagem Bacteriana , Membrana Celular/efeitos dos fármacos , Cromatografia Líquida , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Concentração de Íons de Hidrogênio , Espectrometria de Massas , Dados de Sequência Molecular , Paenibacillus/classificação , Paenibacillus/isolamento & purificação , Paenibacillus/metabolismo , Filogenia , RNA Ribossômico 16S/genética , Rizosfera , Análise de Sequência de DNA , Microbiologia do Solo , Temperatura , Trichophyton/efeitos dos fármacos
15.
Antonie Van Leeuwenhoek ; 99(3): 523-32, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20960060

RESUMO

Prevention or cure of different illnesses through the use of plant latex is a worldwide known concept. The antifungal activity of Hancornia speciosa latex has been observed against Candida albicans. However, H. speciosa latex is not a sterile plant exudate and secondary metabolites produced by bacteria could be involved in fungal inhibition. In the present study, the bacterial communities of the latex from three H. speciosa trees were characterized using traditional plating and molecular methods. Twelve strains isolated from the latex samples were clustered into four groups by amplified ribosomal DNA restriction analysis (ARDRA). One representative of each group was sequenced and they were identified as belonging to the genera Bacillus, Klebsiella, Enterobacter and Escherichia. None of the 12 isolates showed antifungal activity against C. albicans. A lack of a microbial origin for the antifungal properties of latex was noted. DGGE profiles generated from each of the three latex samples showed unique patterns. Sequencing of the DGGE bands demonstrated the affiliation with the genera Klebsiella, Pantoea, Enterobacter and Burkholderia. In addition, clone libraries were generated and the phylogenetic distribution of the 50 analyzed clones was similar to that obtained using DGGE. The presence of some potential pathogens should be considered before using H. speciosa latex in folk medicine.


Assuntos
Apocynaceae/química , Apocynaceae/microbiologia , Bactérias/isolamento & purificação , Bactérias/metabolismo , Látex/farmacologia , Bactérias/classificação , Bactérias/genética , Brasil , Candida albicans/efeitos dos fármacos , DNA Bacteriano/genética , Látex/química , RNA Ribossômico 16S/genética
16.
Mycoses ; 54(5): e474-80, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21554418

RESUMO

The action of the complement system on pigmented and hypopigmented mycelia of the fungus Fonsecaea pedrosoi, the major aetiological pathogen of the chromoblastomycosis is herein discussed. Fungi were grown in medium Czapeck-Dox at 37°C, for 14 days, without shaking to obtain pigmented mycelium. To obtain hypopigmented mycelium, the fungus was grown at the same conditions, but in the dark and with low oxygenation. Activation was measured by complement consumption and enzyme-linked immunosorbent assay. We also observed by immunofluorescence the deposition of C3, C4 fragments and C9 on the surface of the different forms studied. The results indicate that both forms were able to activate the complement system mainly by the alternative pathway. Pigmented mycelia had the highest consumption results, indicating that the pigment, melanin, may have influence in activation.


Assuntos
Ascomicetos/imunologia , Ativação do Complemento , Proteínas do Sistema Complemento/imunologia , Micélio/imunologia , Adulto , Meios de Cultura/química , Escuridão , Ensaio de Imunoadsorção Enzimática , Experimentação Humana , Humanos , Microscopia de Fluorescência , Micologia/métodos , Pigmentos Biológicos/metabolismo
17.
Sci Rep ; 11(1): 20311, 2021 10 13.
Artigo em Inglês | MEDLINE | ID: mdl-34645885

RESUMO

Sulphate-reducing bacteria (SRB) cause fouling, souring, corrosion and produce H2S during oil and gas production. Produced water obtained from Periquito (PQO) and Galo de Campina (GC) onshore oilfields in Brazil was investigated for SRB. Produced water with Postgate B, Postgate C and Baars media was incubated anaerobically for 20 days. DNA was extracted, 16S rDNA PCR amplified and fragments were sequenced using Illumina TruSeq. 4.2 million sequence reads were analysed and deposited at NCBI SAR accession number SRP149784. No significant differences in microbial community composition could be attributed to the different media but significant differences in the SRB were observed between the two oil fields. The dominant bacterial orders detected from both oilfields were Desulfovibrionales, Pseudomonadales and Enterobacteriales. The genus Pseudomonas was found predominantly in the GC oilfield and Pleomorphominas and Shewanella were features of the PQO oilfield. 11% and 7.6% of the sequences at GC and PQO were not classified at the genus level but could be partially identified at the order level. Relative abundances changed for Desulfovibrio from 29.8% at PQO to 16.1% at GC. Clostridium varied from 2.8% at PQO and 2.4% at GC. These data provide the first description of SRB from onshore produced water in Brazil and reinforce the importance of Desulfovibrionales, Pseudomonadales, and Enterobacteriales in produced water globally. Identifying potentially harmful microbes is an important first step in developing microbial solutions that prevent their proliferation.


Assuntos
Microbiota , Campos de Petróleo e Gás , Sulfatos/química , Microbiologia da Água , Biodiversidade , Biofilmes , Biotecnologia , Brasil , DNA Ribossômico/metabolismo , Bases de Dados Genéticas , Desulfovibrionales/genética , Ecologia , Enterobacteriaceae/genética , Gammaproteobacteria/genética , Geografia , Sulfeto de Hidrogênio/química , RNA Ribossômico 16S/genética , Água
18.
Med Mycol ; 48(2): 278-84, 2010 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20141371

RESUMO

Complement activation by spores of Mucor ramosissimus, Mucor plumbeus and Mucor circinelloides was studied using absorbed human serum in the presence or absence of chelators (EGTA or EDTA). We found that the spore caused full complement activation when incubated with EGTA-Mg2+ or without chelators, indicating that the alternative pathway is mainly responsible for this response. In order to compare activation profiles from each species, ELISAs for C3 and C4 fragments, mannan binding lectin (MBL), C-reactive protein (CRP) and IgG studies were carried out. All proteins were present on the species tested. Immunofluorescence tests demonstrated the presence of C3 fragments on the surface of all samples, which were confluent throughout fungal surfaces. The same profile of C3, C4, MBL, CRP and IgG deposition, observed in all species, suggests a similar activation behavior for these species.


Assuntos
Ativação do Complemento/imunologia , Mucor/fisiologia , Esporos Fúngicos/imunologia , Proteína C-Reativa/metabolismo , Complemento C3/metabolismo , Complemento C4/metabolismo , Ácido Edético , Ácido Egtázico , Ensaio de Imunoadsorção Enzimática , Técnica Direta de Fluorescência para Anticorpo , Humanos , Imunoglobulina G/metabolismo , Lectina de Ligação a Manose/metabolismo , Mucor/imunologia
19.
Parasitol Res ; 106(5): 1249-52, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20177905

RESUMO

Parasites from the genus Leishmania cause a variety of disease states in humans and other mammals in tropical and subtropical regions, which include cutaneous, mucocutaneous and visceral leishmaniasis. The elaboration of a culture medium for the in vitro cultivation of Leishmania spp., which promotes the growth and differentiation of the parasites, is an important tool for diagnosis, biochemical, biological and immunological studies in the genus. Herein, we have reported the development of a rapid, inexpensive and reliable monophasic culture medium. The novel medium, designated PBHIL, promoted an excellent parasite growth, generating high quantities of promastigotes with long-term viability, and was able to induce cellular differentiation of L. amazonensis promastigotes to the amastigote-like forms (93%). Additionally, we reported the influence of this novel medium on the biochemical characteristics of L. amazonensis and on the interaction of this parasite parasites with mammalian macrophages.


Assuntos
Meios de Cultura/química , Leishmania/crescimento & desenvolvimento , Parasitologia/métodos , Animais , Diferenciação Celular , Sobrevivência Celular , Células Cultivadas , Feminino , Leishmania/citologia , Macrófagos Peritoneais/parasitologia , Camundongos , Camundongos Endogâmicos BALB C
20.
Mycopathologia ; 170(1): 31-7, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20195904

RESUMO

Fonsecaea pedrosoi is the major etiologic agent of chromoblastomycosis. The virulence of F. pedrosoi is a meagerly explored phenomenon. The ability to interact with host cells and the production of hydrolytic enzymes are thought to be important virulence mechanisms of fungal pathogens. Here, we measured the production of two distinct lipolytic enzymes, phospholipase and esterase, by three clinical strains of F. pedrosoi isolated from chromoblastomycosis lesions, as well as their capabilities to interact with epithelial cells. All the strains were excellent esterase producers, generating elevated hydrolytic halos after 5 days of growth. Conversely, phospholipase activity was detected only after 10 days, except for the most recent strain of F. pedrosoi (Magé) in which measurable phospholipase activity was detected on day 5. The ability to interact with epithelial cells was also investigated. Regarding the adhesion capability, an indirect connection was observed in relation to the adaptation time of each strain in axenic culture, in which Magé strain showed the best adhesion ability followed by LDI 11428 and 5VPL strains. Both 5VPL and Magé strains were also detected inside the epithelial cells, while the LDI 11428 strain was rarely detected in cytoplasmatic vacuolar compartments. Moreover, these F. pedrosoi strains were able to cause injury in epithelial cells.


Assuntos
Ascomicetos/enzimologia , Cromoblastomicose/microbiologia , Células Epiteliais/microbiologia , Esterases/metabolismo , Proteínas Fúngicas/metabolismo , Fosfolipases/metabolismo , Fatores de Virulência/metabolismo , Animais , Ascomicetos/isolamento & purificação , Ascomicetos/patogenicidade , Células CHO , Adesão Celular , Cricetinae , Cricetulus , Humanos , Vacúolos/microbiologia
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