Susceptibility of virulent and resistant Escherichia coli strains to non-polar and polar compounds identified in Microplumeria anomala.

BACKGROUND AND AIM: Escherichia coli is one of the main pathogens responsible for veterinary and human infections, and it is associated with significant economic losses in the livestock, as it causes severe diseases to humans, particularly in children. For that reason, there is a need for introducing new drugs to treat E. coli diseases. The Brazilian species richness is a source of potential new antibacterial natural products. The study aimed at the biological and chemical investigation of the organic extract obtained from the stem of Microplumeria anomala (Apocynaceae), EB127, as it was identified as a potential source of new antibacterial compounds to be used in Veterinary. MATERIALS AND METHODS: The antibacterial activity was evaluated by disk diffusion and microdilution assays; chromatography, nuclear magnetic resonance spectrometry, and mass spectrometry were used in the isolation and identification of compounds. RESULTS: EB127 showed activity against E. coli ATCC25922, and against three E. coli strains that were isolated from frigarte's cloaca, named 31/1A, 35A, and 51A. Lupeol, 3-acetyl-11-oxo-ß-amyrin, 3-acetyl-11-oxo-α-amyrin, sitosterol, stigmasterol, 3ß,7α-dihydroxy-cholest-5-ene, 3ß-hydroxy-cholest-5-en-7-one, and 3ß-hydroxy-cholest-5,22-dien-7-one were identified in fraction Hex/CHCl3, while loganin, loganic acid, methylanomaline, and anomaline were all identified in EB127 and protocatechuic acid hexoside, ferulic acid, secoxyloganin, feruloylquinic acid, vanillic acid hexoside, protocatechuic acid-4-O-ß-hexoside, and rosmarinic acid were tentatively identified in fraction 10%ACN/H2O. E. coli 51A (virulent/non-resistant) showed sensitivity to the antibacterial action of fraction Hex/CHCl3 which contains alkaloids, triterpenes, and steroids, while E. coli 35A (resistant/non-virulent) were more susceptible to 10%ACN/H2O, which contains iridoids as loganin and loganic acid, and glycosylated and non-glycosylated caffeic acids. CONCLUSION: Fraction 10%ACN/H2O is of interest in pursuing new drugs to treat resistant E. coli, in veterinary. All compounds were isolated from the plant for the first time and have shown potential as new antibacterial natural products from Amazon plants to be used in veterinary and human diseases.

Amazon climatic factors driving terpene composition of Iryanthera polyneura Ducke in terra-firme forest: A statistical approach.

Iryanthera polyneura (Myristicaceae) is popularly known as cumala-colorada, and can be found in the Amazon rain forest. The present study aimed the evaluation of the chemical composition of the essential oils (EOs) obtained from the leaves of I. polyneura throughout a two-year period in order to correlate chemical findings with seasonality and climatic variation. Leaves from I. polyneura were collected 15 times from three different individuals, identified as 22EO, 80EO and 530EO, between the years of 2009 and 2011. The EOs were obtained and tested by gas-chromatography-mass spectrometry techniques. ANOVA and multivariate analyses were used to assess the relationship between the percentage of terpenes and seasonal/climatic variations. Fifty-nine compounds were detected in the EOs, of which 44 were identified (74.5%). The main components of the EOs were spathulenol (6.42 ± 1.02%), α-cadinol (5.82 ± 0.40%) and τ-muurolol (5.24 ± 0.03%). Higher levels of spathulenol were observed in 22EO during rainy season, while τ-muurolol occurred in high amounts during the dry season, as did α-cadinol in 22EO and 80EO. Correspondence analysis revealed a distinction between two groups of EOs based on seasonality, whereas a canonic correspondence analysis and ANOVA determined how the major compounds were related to both seasonality and climatic factors. Non-metric multidimensional scaling in association with an analysis of similarities showed that the abundance and composition of terpenes in the samples varied moderately among the three Iryanthera individual plants. Present findings have shown that there is variation in the occurrence of the major compounds spathulenol, τ-muurolol and α-cadinol produced by I. polyneura and that the pattern of variation is related to season and climatic changes.

The Synergistic Behavior of Antioxidant Phenolic Compounds Obtained from Winemaking Waste's Valorization, Increased the Efficacy of a Sunscreen System.

Grape pomace retains polyphenols in the peels and in the seeds after winemaking, which is indicative of the high valorization potential of this industrial waste. There is strong evidence that phenolics are robust antioxidants and confer photoprotection; thus, it is rational to apply these active compounds from winemaking waste to sunscreens, in order to increase UV protection. Despite the importance of this class of cosmetics to public health, more efficacious strategies are still needed to overcome the problems caused by the photoinstability of some UV filters. The hydroethanolic extract of Vitis vinifera L. grapes was obtained by percolation and then lyophilized. Six formulations were developed: Type I-cosmetic base and UV filters; Type II-cosmetic base and extract; and Type III-cosmetic base, extract and UV filters. Each formulation was prepared in the pHs 5 and 7. The antioxidant activities of the samples were measured by DPPH• and expressed in Trolox® equivalents (TE), and their photostability and in vitro sun protection factor (SPF) were analyzed by diffuse reflectance spectrophotometry. The anti-radical efficiencies observed in the formulations with grape extract were: (II) 590.12 ± 0.01 µmol TE g-1 at pH 5 and 424.51 ± 0.32 µmol TE g-1 at pH 7; (III) 550.88 ± 0.00 µmol TE g-1 at pH 5 and 429.66 ± 0.10 µmol TE g-1, at pH 7, demonstrating that the UV filters, butylmethoxydibenzoyl methane, ethylhexyl methoxycinnamate and ethylhexyl dimethyl 4-aminobenzoic acid had no influence on this effect. The photoprotective efficacy and the photostability of formulation III containing the extract and UV filters at pH 5 suggested that a synergism between the active molecules provided an 81% increase in SPF. Additionally, this was the only sample that maintained a broad spectrum of protection after irradiation. These results confirmed that the grape pomace extract has multifunctional potential for cosmetic use, mainly in sunscreens, granting them superior performance.

Preliminary analysis of Brazilian Amazonian plant extracts using bench-top assays in a through-put basis aiming the identification of potential antioxidant natural product

Plants are a source of compounds that are used for the treatment of human and veterinary diseases. Brazil is one of the richest countries in the world in terms of biodiversity. The present study evaluated extracts using thin-layer chromatography to identify antioxidant activity and determine the presence of groups of compounds, in a large-scale basis. A total of 1,260 aqueous and organic extracts were obtained from plants that were collected in the Amazon and Atlantic rain forests. Thin-layer chromatography was performed to evaluate the presence of alkaloids, anthraquinones, cardioactive glicosides, terpenes, and phenolics and determine antioxidant and radical scavenging activity using the following reagents: Dragendorff's reagent, KOH, Kedde's reagent, sulphuric acid, NP reagent, ß-carotene, and 2,2-diphenyl-1-picrylhydrazyl (DPPH). Of the 1,260 extracts, 837 (66.43%) presented a ß-carotene/bleaching response, and 1,205 (95.63%) presented a radical scavenging response. Alkaloids were found in 203 extracts (16.11%). Anthraquinones were found in 14 extracts (1.11%), cardenolides were found in eight extracts (0.63%). The present findings shows the importance of the Brazilian Amazon plants as sources of antioxidant and radical scavenging active compounds.(AU)

Can mouth washes containing chlorhexidine 0.12% be used as synonym of a water solution of chlorhexidine 0.12%? / Pode lavagens da boca contendo 0.12 % de clorexidina ser utilizado como sinónimo de uma solução aquosa de cloro-hexidina 0.12 % ?

Chlorhexidine digluconate (CHX) is a gold standard drug in dentistry and is widely used as a reference in both in vitro and in vivoexperiments. Due to ease of access, mouth washes containing CHX 0.12% are used as a substitute for aqueous CHX 0.12% solution in laboratory experiments. Additionally, it is well known that for product flavor purposes, volatile compounds are added to mouth washes formulations. Volatiles added to CHX 0.12% may improve wash's antibacterial ability. Volatiles add potency to the mouth wash formulation. Compared with an aqueous CHX 0.12% solution, it is proposed that CHX solutions and Periogard^® would have antimicrobial activity. Antimicrobial activity was assessed in the present study via disk diffusion assays against Streptococcus mutans, Streptococcus sanguinisand Escherichia coli. Periogard^® showed a significantly higher antibacterial activity in relation to CHX 0.12% (p<0.05) and a similar activity in relation to CHX 1% (p>0.05). Periogard^(r) volatiles were analyzed by gas-chromatography/mass spectrometry (GCMS) and the presence of antibacterial menthol, menthone, isomenthol, menthyl acetate, trans-anethol and eugenol was verified. Finally, the use of Periogard^® as a synonym of CHX 0.12% must be avoided, because its antibacterial activity is closely related to CHX 1%...

Gluconato de clorexidina (CHX) é um fármaco considerado padrão ouro, em Odontologia, amplamente usado como referência em estudos in vitro e in vivo. Em razão da facilidade de acesso, enxaguatórios bucais que contêm CHX 0,12% são usados em substituição à solução aquosa de clorexidina (CHX 0,12%), em experimentos laboratoriais. É sabido que devido à palatabilidade do produto, os mesmos enxaguatórios bucais contêm compostos voláteis em sua formulação, além da CHX 0.12%. Visto que voláteis adicionados podem acrescentar poder antibacteriano à formulação, a comparação da resposta antibacteriana da solução aquosa de CHX em diferentes concentrações e de Periogard^® é proposta no presente artigo. Para tanto, utilizou-se o ensaio do disco de difusão em ágar com inóculos de Streptococcus mutans, Streptococcus sanguinis e Escherichia coli. Periogard^® mostrou atividade antibacteriana significativa contra as três cepas analisadas, quando comparada à atividade de CHX 0.12% (p<0,05) e atividade similar à CHX diluída a 1% (p>0,05). A presença de compostos voláteis no Periogard^® foi analisada por GC-MS e observou-se que mentol, mentona, isomentol, acetato de mentila, trans-anetol e eugenol estão presentes na formulação. Deste modo, o uso de Periogard^(r) como sinônimo de CHX 0,12% deve ser evitado, uma vez que sua atividade se assemelha àquela da CHX diluída a 1%...

In vitro activity of Amazon plant extracts against Enterococcus faecalis.

Previous studies analyzing 2,200 plant extracts indicated anti-enterococcal activity in 25 extracts obtained from Brazilian forests' plants. In the present study, these extracts were subjected to microdilution broth assay (MDBA) and disk diffusion assay (DDA) using planktonic Enterococcus faecalis ATCC(®) 29212™ and were submitted to phytochemical analysis in TLC and HPLC. Three extracts obtained from Ipomoea alba (MIC < 40 µg/mL), Diclinanona calycina (MIC ≤ 40 µg/mL) and Moronobea coccinea (40 < MIC < 80 µg/mL; MBC = 80 µg/mL) showed significant bactericidal activity in the MDBA and four extracts obtained from I. alba (14.04 ± 0.55 mm diameter) S. globulifera (14.43 ± 0.33 mm and 12.18 ± 0.28 mm diameter) and Connarus ruber var. ruber (13.13 ± 0.18 mm diameter) were active in DDA. Residues H2O obtained from Psidium densicomum (mean of 16.78 mm diameter) and from Stryphnodendron pulcherrimum (mean of 15.97 mm diameter) have shown an improved antibacterial activity after fractionation if compared to that obtained from the respective crude extracts. Antioxidant activity was observed in some residues of the active extracts. TLC analysis showed that phenolic compounds are likely to be found in active extracts. Three molecules were isolated from S. globulifera and were identified by (13)C NMR lupeol, α-amyrin and 3ß-hydroxyglutin-5-ene. The present chemical and biological findings suggest that these extracts are a potential source of new anti-Enterococcus compounds to be introduced in endodontic therapy.

In vitro activity of Amazon plant extracts against Enterococcus faecalis

Previous studies analyzing 2,200 plant extracts indicated anti-enterococcal activity in 25 extracts obtained from Brazilian forests' plants. In the present study, these extracts were subjected to microdilution broth assay (MDBA) and disk diffusion assay (DDA) using planktonic Enterococcus faecalis ATCC® 29212TM and were submitted to phytochemical analysis in TLC and HPLC. Three extracts obtained from Ipomoea alba (MIC < 40 µg/mL), Diclinanona calycina (MIC < 40 µg/mL) and Moronobea coccinea (40 < MIC < 80 µg/mL; MBC = 80 µg/mL) showed significant bactericidal activity in the MDBA and four extracts obtained from I. alba (14.04 ± 0.55 mm diameter) S. globulifera (14.43 ± 0.33 mm and 12.18 ± 0.28 mm diameter) and Connarus ruber var. ruber (13.13 ± 0.18 mm diameter) were active in DDA. Residues H2O obtained from Psidium densicomum (mean of 16.78 mm diameter) and from Stryphnodendron pulcherrimum (mean of 15.97 mm diameter) have shown an improved antibacterial activity after fractionation if compared to that obtained from the respective crude extracts. Antioxidant activity was observed in some residues of the active extracts. TLC analysis showed that phenolic compounds are likely to be found in active extracts. Three molecules were isolated from S. globulifera and were identified by 13C NMR lupeol, α-amyrin and 3β-hydroxyglutin-5-ene. The present chemical and biological findings suggest that these extracts are a potential source of new anti-Enterococcus compounds to be introduced in endodontic therapy.

Anti-Streptococcal activity of Brazilian Amazon Rain Forest plant extracts presents potential for preventive strategies against dental caries

Caries is a global public health problem, whose control requires the introduction of low-cost treatments, such as strong prevention strategies, minimally invasive techniques and chemical prevention agents. Nature plays an important role as a source of new antibacterial substances that can be used in the prevention of caries, and Brazil is the richest country in terms of biodiversity. Objective: In this study, the disk diffusion method (DDM) was used to screen over 2,000 Brazilian Amazon plant extracts against Streptococcus mutans. Material and Methods: Seventeen active plant extracts were identified and fractionated. Extracts and their fractions, obtained by liquid-liquid partition, were tested in the DDM assay and in the microdilution broth assay (MBA) to determine their minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs). The extracts were also subjected to antioxidant analysis by thin layer chromatography. Results: EB271, obtained from Casearia spruceana, showed significant activity against the bacterium in the DDM assay (20.67±0.52 mm), as did EB1129, obtained from Psychotria sp. (Rubiaceae) (15.04±2.29 mm). EB1493, obtained from Ipomoea alba, was the only extract to show strong activity against Streptococcus mutans (0.08 mg/mL<MIC<0.16 mg/mL; MBC=0.16 mg/mL) in the MBA. Conclusions: The active extracts, discovered in the Amazon rain forest, show potential as sources of new antibacterial agents for use as chemical coadjuvants in prevention strategies to treat caries. .

Anti-Streptococcal activity of Brazilian Amazon Rain Forest plant extracts presents potential for preventive strategies against dental caries.

UNLABELLED: Caries is a global public health problem, whose control requires the introduction of low-cost treatments, such as strong prevention strategies, minimally invasive techniques and chemical prevention agents. Nature plays an important role as a source of new antibacterial substances that can be used in the prevention of caries, and Brazil is the richest country in terms of biodiversity. OBJECTIVE: In this study, the disk diffusion method (DDM) was used to screen over 2,000 Brazilian Amazon plant extracts against Streptococcus mutans. MATERIAL AND METHODS: Seventeen active plant extracts were identified and fractionated. Extracts and their fractions, obtained by liquid-liquid partition, were tested in the DDM assay and in the microdilution broth assay (MBA) to determine their minimal inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs). The extracts were also subjected to antioxidant analysis by thin layer chromatography. RESULTS: EB271, obtained from Casearia spruceana, showed significant activity against the bacterium in the DDM assay (20.67±0.52 mm), as did EB1129, obtained from Psychotria sp. (Rubiaceae) (15.04±2.29 mm). EB1493, obtained from Ipomoea alba, was the only extract to show strong activity against Streptococcus mutans (0.08 mg/mL

New trends in dentistry: plant extracts against Enterococcus faecalis. The efficacy compared to chlorhexidine

Enterococcus faecalis is an important pathogen associated with endodontic diseases, and its elimination and control are of paramount importance, as it represents one of the major causes of failure in the treatment of endodontic disease. Twenty-five plant extracts obtained from Brazilian forests were found to be effective against planktonic E. faecalis and were subjected to two traditional antibacterial assays, the microdilution broth assay (MDBA) and the disk diffusion assay (DDA), using chlorhexidine (CHX) as a control. Seven out of 25 extracts showed significant antibacterial activity and were tested in a biofilm assay, and three of these extracts were subjected to chemical fractionation. Residues were tested for their antibacterial activity, and the first chemical findings were described based on thin layer chromatography (TLC). Extracts obtained from Ipomoea alba, Symphonia globulifera and Moronobea coccinea showed significant bactericidal activity in the MDBA. The same I. alba and S. globulifera extracts, as well as the extract obtained from Connarus ruber var. ruber, showed significant activity in the DDA. RH2O obtained from Psidium densicomum and Stryphnodendron pulcherrimum showed better antibacterial activity compared to the respective crude extracts and CHX. TLC analysis showed that phenolic compounds and triterpenes represent the first findings of chemical groups that may occur in all species. The results of the present study include the discovery of six active extracts against planktonic E. faecalis and support further testing via assays involving biofilm formation, as well as the determination of the compounds' chemical profiles, as their activity was significantly better than that observed for CHX.

New trends in dentistry: plant extracts against Enterococcus faecalis. The efficacy compared to chlorhexidine.

Enterococcus faecalis is an important pathogen associated with endodontic diseases, and its elimination and control are of paramount importance, as it represents one of the major causes of failure in the treatment of endodontic disease. Twenty-five plant extracts obtained from Brazilian forests were found to be effective against planktonic E. faecalis and were subjected to two traditional antibacterial assays, the microdilution broth assay (MDBA) and the disk diffusion assay (DDA), using chlorhexidine (CHX) as a control. Seven out of 25 extracts showed significant antibacterial activity and were tested in a biofilm assay, and three of these extracts were subjected to chemical fractionation. Residues were tested for their antibacterial activity, and the first chemical findings were described based on thin layer chromatography (TLC). Extracts obtained from Ipomoea alba, Symphonia globulifera and Moronobea coccinea showed significant bactericidal activity in the MDBA. The same I. alba and S. globulifera extracts, as well as the extract obtained from Connarus ruber var. ruber, showed significant activity in the DDA. RH2O obtained from Psidium densicomum and Stryphnodendron pulcherrimum showed better antibacterial activity compared to the respective crude extracts and CHX. TLC analysis showed that phenolic compounds and triterpenes represent the first findings of chemical groups that may occur in all species. The results of the present study include the discovery of six active extracts against planktonic E. faecalis and support further testing via assays involving biofilm formation, as well as the determination of the compounds' chemical profiles, as their activity was significantly better than that observed for CHX.

Cytotoxicity of cashew flavonoids towards malignant cell lines.

The leaves of the Cashew plant (Anacardium occidentale L.) are used by the folk medicine in South America and West Africa. This plant is rich in flavonoids, which are polyphenolic compounds widespread in plants, and that have diverse physiological effects. In a sub-acute toxicity assay it was found that an ethanolic extract of Cashew leaves elicited lymphopenia in rats. The extract was also found to be cytotoxic and to induce apoptosis in Jurkat (acute lymphoblastic leukemia) cells. The crude ethanolic extract was fractionated and resolved by HPLC. One of the four fractions obtained led to the isolation of the biflavonoid agasthisflavone. [(3)H]-thymidine incorporation assays and flow cytometry analysis showed that the isolated compound displayed a high anti-proliferative effect in Jurkat cells with an IC(50) of 2.4 µg/ml (4.45 µM). The effect of agathisflavone on the acute promyelocytic leukemia cell line HL60, Burkitt lymphoma Raji cells and Hep-2 laryngeal carcinoma cells was also tested. The two latter ones were only mildly affected by agathisflavone. It is also shown that agathisflavone induces apoptosis in Jurkat cells and it this proposed that this is the likely mechanism of agathisflavone specific cytotoxicity.

Evaluation of gastroprotective activity of Plinia edulis (Vell.) Sobral (Myrtaceae) leaves in rats.

Plinia edulis, an arboreous species popularly known as "cambucá", is native to the Brazilian Atlantic Rain Forest. Despite its traditional uses, no reports are available on the safety of this utilization or on the relationship between the antiulcer activity of its extract and its phytochemical compounds. This paper reports on the investigation of the acute toxicity and gastroprotective effect of the aqueous ethanol extract of leaves of Plinia edulis on HCl/ethanol-induced ulcers. In order to correlate the secondary metabolites and the efficacy of the crude drug in traditional medicine, the extract was submitted to chromatographic fractionation after solvent partition. The extract did not show acute toxicity in mice treated with 5 g/kg p.o., but exhibited significant antiulcer activity in rats at doses of 100, 200, and 400mg/kg p.o., more active than the reference drug lansoprazole.The ethyl acetate fraction yielded beta-amyrin, oleanolic acid, ursolic acid, and maslinic acid, which were identified based on spectrometric analyses. Since antiulcerogenic activity is not restricted to one class of compounds in plants, the triterpenoids isolated in the extract can be associated with the observed effect.