HEPATOPROTECTIVE POTENTIAL OF EUGENIA?UNIFLORA?L AGAINST GENTAMYCIN INDUCED HEPATOXICITY

Hepatotoxicity is a common drug adverse effect and gentamycin has been linked to hepatotoxic adverse reactions. Folklore and ethnobotanical studies indicate Eugenia uniflora L is used in the treatment of gastrointestinal ailments and has exhibited diverse biological activities. We investigated the hepatoprotective potential of this plant in treating gentamycin-induced hepatoxicity and compared the effect with Celebrex a COX2 selective inhibitor. Twenty-eight male adult Wistar rats average of 225g were used for the study and randomised into groups as described: normal control( normal saline), negative control: Gentamycin (40mg/kg. i.p), positive control; Gentamycin (40mg/kg. i.p) +5mg/kg Celebrex. Extract low dose: Gentamycin (40mg/kg i.p)+(50mg/kg) Eugenia uniflora L. leaves, intermediate-dose: Gentamycin (40mg/kg i.p) +100mg/kg Eugenia uniflora L leaves, high dose : Gentamycin (40mg/kg, i.p)+ (200mg/kg, ) of Eugenia uniflora L leaves. Our findings indicate that the body weight was unaffected throughout the experiment, as clearly demonstrated by the lack of significant variability (p<0.05). Hepatotoxicity was confirmed by dose-dependent alteration in Liver marker enzymes, including AST, ALT, and ALP. Eugenia uniflora L leaves were able to ameliorate the levels of these liver enzymes to a normal level. Liver tissue revealed a dose-dependent curative effect with Eugenia uniflora L compared to the COX2 inhibitor (Celebrex) treatment. Consequently, we hereby report, for the first time, that an aqueous extract of Eugenia uniflora L leaves confers hepatoprotection against gentamycin-induced hepatoxicity in Wistar rats.

In vivo Chemosuppressive effects of Combinations of Three Nigerian Ethnomedicinal Antimalarial Plants

Aims: Appropriate ratios for combining some African ethnomedicinal plants with proven anti-plasmodial activity were determined with the aim of obtaining herbal remedies with higher efficacies. Place and Duration of the Study: Study was conducted in the Department of Pharmacognosy, Obafemi Awolowo University, Ile-Ife Nigeria. Study Design: Infusions of dried powders of three Nigerian ethnomedicinal plants, Eugenia uniflora leaf, Gongronema latifolium root and stem and Artocarpus altilis stem bark, were separately evaluated and in varied combination ratios for their anti-plasmodial activities Methodology: The infusions were separately prepared, concentrated in vacuo, freeze-dried and evaluated at 400mg/kg against Plasmodium berghei berghei parasites, using the Peter’s four-day chemosuppressive mice model. Normal saline and chloroquine (10mg/kg) were negative and positive controls, respectively. 2- and 3-combinations of the infusions were similarly tested. Results: Of the 2-plant combination ratios, 1:1, 1:3, 2:3 of EG and GA elicited percentage chemo suppressions which were comparable (P=.28; P=.07) to those of the single drugs. Only the EG ratios gave activities that were comparable (P=.28) to the positive control in addition to double survival times and high survivor values by EG 2:3 and GA 1:3. However, those of the EA group, gave relatively low values, barely above 20% with only the ratios 2:3 and 3:2 giving values which were significantly (P=.00) higher than negative control with double survival times. The 3-plant combination ratios, EGA 2:1:2 and 3:3:2 gave suppressions that were significantly (P=.00) higher than the negative control with others but comparable (P=.33) activities to those of the individual drugs. The other ratios with low suppression values were relatively inactive. But three ratios, EGA 3:1:2, 2:1:1, 1:1:1 elicited survival times doubled (204, 242 and 202 %) that of the negative control without commensurate high antiplasmodial activities. Conclusion: Ethnomedicinal antimalarial plants should not be combined without a data of previous scientific evaluations.

Essential oils in pathogen resistance induction of Eucalyptus benthamii Maiden et Cambage / Óleos essenciais na indução de resistência a patógenos em Eucalyptus benthamii Maiden et Cambage

ABSTRACT: This study evaluated the essential oils of Melaleuca alternifolia, Casearia sylvestris and Eugenia uniflora as inducers of defense mechanisms in Eucalyptus benthamii seedlings. Four mL of each oil, with a concentration of 0.75% were sprayed in E. benthamii seedlings and two bioassays were performed, in the first, the essential oils were sprayed and after 30 days, sugars, proteins, peroxidases, phenylalanine ammonia (PAL), and phenols were evaluated and; in the second, seven days after the first analysis, the essential oils were again sprayed and after three days, the same variables were evaluated. The essential oils of M. alternifolia, C. sylvestris, and E. uniflora sprayed had no significant effects on E. benthamii seedlings after 30 days in terms of total sugars, proteins, peroxidase, PAL activity, and phenols. However, when M. alternifolia and E. uniflora essential oils were sprayed seven days after the first analysis with evaluation after 3 days, an increase in total sugars was observed. After these days, all essential oils promoted an increase in protein levels. The oils of E. uniflora and C. sylvestris also increased peroxidase levels. The PAL defense enzyme not showed increased when essential oils were used. The essential oils of M. alternifolia and C. sylvestris had potential as inducers of defense mechanisms on E. benthamii seedlings after 3 days of their application, what it demonstrated not be permanent.

RESUMO: Este trabalho teve como objetivo avaliar os óleos essenciais de Melaleuca alternifolia, Casearia sylvestris e Eugenia uniflora como indutores de mecanismos de defesa em mudas de Eucalyptus benthamii. Quatro mL de cada óleo, com concentração de 0,75%, foram pulverizados em mudas de E. benthamii e foram realizados dois bioensaios. No primeiro foram pulverizados os óleos essenciais e após 30 dias, açúcares, proteínas, peroxidases, fenilalanina amônia (PAL) , e fenóis foram avaliados e; na segunda, sete dias após a primeira análise, os óleos essenciais foram novamente pulverizados e após três dias, as mesmas variáveis foram avaliadas. Os óleos essenciais de M. alternifolia, C. sylvestris e E. uniflora pulverizados não tiveram efeito significativo sobre as mudas de E. benthamii após 30 dias em termos de açúcares totais, proteínas, peroxidase, atividade PAL e fenóis. Entretanto, quando os óleos essenciais de M. alternifolia e E. uniflora foram pulverizados sete dias após a primeira análise com avaliação após três dias, foi observado um aumento nos açúcares totais. Depois desses dias, todos os óleos essenciais promoveram aumento nos níveis de proteínas. Os óleos de E. uniflora e C. sylvestris também aumentaram os níveis de peroxidase. A enzima de defesa PAL não se mostrou aumentada quando óleos essenciais foram usados. Os óleos essenciais de M. alternifolia e C. sylvestris apresentaram potencial como indutores de mecanismos de defesa em mudas de E. benthamii após três dias de sua aplicação, o que demonstrou não ser permanente.

Antioxidant activity, antibacterial and inhibitory effect of intestinal disaccharidases of extracts obtained from Eugenia uniflora L. seeds / Atividade antioxidante, antibacteriana e efeito inibidor de dissacaridases intestinais de extratos obtidos das sementes de Eugenia uniflora L

The use of medicinal plants for disease prevention, treatment and cure is an ancient practice used by humanity, and many plants species are used in bioprospecting research. In this context, its stands out Eugenia uniflora L., populary known as pitangueira and belongs to the Myrtaceae family, with a wide geographic distribution and native of Brazil. In view of the therapeutic qualities of the plant and the lack of the studies on its seeds, the present study had as objective to evaluate the phytochemical profile of the extracts of Eugenia uniflora L. seeds, from different solvents, as well as their antibacterial activity, antioxidant and its inhibitory effect of intestinal disaccharidases. Results showed a high content of phenolic compounds and total flavonoids, thus characterizing antioxidant activity, also highlighting the best bacteriostatic action for the Gram positive strain of Staphylococcus aureus in the ethanolic fraction. Regarding the disaccharidases, a strong inhibitory action was observed for all concentrations, evidencing a antihyperglycemic potential. The present research allowed to concluded that Eugenia uniflora L. seeds have promising biological activities for the industrial sector, but a more detailed investigation is needed regarding their bioactive compounds.

A utilização de plantas com fins medicinais para prevenção, tratamento e cura de doenças é uma prática antiga utilizada pela humanidade, sendo que muitas espécies vegetais são usadas para a pesquisa da bioprospecção. Neste contexto, destaca-se a Eugenia uniflora L., conhecida popularmente como pitangueira e pertencente à família Myrtaceae, com ampla distribuição geográfica e nativa do Brasil. Diante das qualidades terapêuticas da planta e a falta de estudos sobre suas sementes, o presente trabalho teve como objetivo avaliar o perfil fitoquímico dos extratos das sementes de Eugenia uniflora L. a partir de diferentes solventes, bem como sua atividade antibacteriana, antioxidante e seu efeito inibidor de dissacaridases intestinais. Os resultados mostraram um alto teor de compostos fenólicos e flavonóides totais, caracterizando a atividade antioxidante, destacando também a melhor ação bacteriostática para a cepa Gram positiva de Staphylococcus aureus na fração etanólica. Em relação às dissacaridases, uma forte ação inibitória foi observada para todas as concentrações, evidenciando um potencial anti-hiperglicêmico. A presente pesquisa permitiu concluir que as sementes de Eugenia uniflora L. apresentam atividades biológicas promissoras para o setor industrial, mas é necessária uma investigação mais detalhada de seus compostos bioativos.

Chemical composition of various plant extracts and their in vitro efficacy in control of Fasciola hepatica eggs / Composição química de vários extratos vegetais e sua eficácia in vitro no controle de ovos de Fasciola hepática

ABSTRACT: Fasciolosis has been diagnosed in cattle, goats, sheep and horses in southern and southeastern Brazil. Effective alternative treatments are the targets of study. One promising alternative is the use of plant extracts. The aim of this study was to perform phytochemical analysis of extracts of Eugenia uniflora L., Harpagophytum procumbens, Psidium guajava L. and Stryphnodendron adstringens, and to evaluate the in vitro efficacy of these extracts on ovicidal activity in Fasciola hepatica. Plant extracts were analyzed for phytochemical properties. F. hepatica eggs were collected directly from the gallbladders of animals diagnosed as positive for fasciolosis on post mortem examination. One hundred eggs were incubated with 3 ml of each extract at concentrations of 0.10%, 0.25% and 0.50%, albendazole 0.50% (positive control) or tap water (negative control). To determine anti larval efficacy of each plant extract, hatched eggs were counted and the averages were used. Phytochemical analysis revealed the presence of phenolic compounds, tannins and terpenes in most extracts. E. uniflora L. extract was 100% effective at 0.10%, H. procumbens was effective at 0.25% and P. guajava L. and S. adstringens extracts were 100% effective at all concentrations tested. Taken together, the data suggested that ovicidal activity in F. hepatica is due to the presence of these bioactive compounds.

RESUMO: A fasciolose tem sido diagnosticada em bovinos, caprinos, ovinos e equinos no sul e sudeste do Brasil, sendo que tratamentos alternativos mais eficazes são alvos de estudo. Umas das alternativas promissoras é o uso de extratos vegetais no controle dessa e outras enfermidades. O objetivo deste estudo foi realizar a análise fitoquímica dos extratos de Eugenia uniflora L., Harpagophytum procumbens, Psidium guajava L. e Stryphnodendron adstringens, além de avaliar a eficácia in vitro desses extratos na atividade ovicida em Fasciola hepatica. Os extratos vegetais foram obtidos e analisados para determinação fitoquímica. Ovos de F. hepatica foram coletados diretamente das vesículas biliares de animais diagnosticados como positivos para fasciolose no exame post mortem. Cem ovos foram incubados com três mililitros de cada extrato nas concentrações de 0,10%, 0,25% e 0,50%, de albendazol a 0,50% (controle positivo) e água de torneira (controle negativo). Para determinar a eficácia de cada extrato vegetal os ovos eclodidos foram contados, e a média utilizada para os cálculos de eficácia. A análise fitoquímica revelou a presença de compostos fenólicos, taninos e terpenos na maioria dos extratos. O extrato de E. uniflora L. apresentou eficácia de 100% na concentração de 0,10%, o de H. procumbens a 0,25% e os extratos de P. guajava L. e S. adstringens apresentaram 100% de eficácia em todas as concentrações testadas. Assim, sugere-se que a atividade ovicida em F. hepatica seja devido à presença desses compostos bioativos.

Anti-Helicobacter pylori and Anti-inflammatory Properties of Eugenia uniflora L

Abstract Helicobacter pylori is a bacterium that reaches half of the world population and it's recognized as the main cause of chronic gastritis and peptic ulcer. In this study, we evaluated the anti-H. pylori, antioxidant and immunomodulatory activities of the methanolic (MeOH) extract of Eugenia uniflora leaves and chemical profile. Anti-H. pylori activity was evaluated by spectrophotometric broth microdilution technique by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), in addition to the evaluation of the effect on the urease enzyme. The antioxidant activity was evaluated by capturing O2 •-, HOCl e NO• radicals. The immunomodulatory effect was evaluated on the cytokines TNF-α, IL-6 and on nitric oxide through inhibition in LPS-stimulated macrophages. The chemical profile was performed by total phenolic, tannin and flavonoid contents and mass spectrometry analysis by ESI-FT-ICR MS. In the anti-H. pylori assay the extract showed MIC of 128 μg/mL, however it did not obtain MBC. The extract also showed ability to inhibit the urease enzyme about 20%. The antioxidant activity of the MeOH extract showed EC50 values of 29.77 µg/mL, 15.71 µg/mL and 442.10 µg/mL to O2 •-, HOCl and NO•, respectively. The extract also showed influence on the release of TNF-α, IL-6 and NO in LPS-stimulated macrophages, ranging from 39% to 97% inhibition. Flavonoids, phenylpropanoids, tannins, triterpenoids and carbohydrates were the major classes of compounds present in the MeOH extract as identified by (-)-ESI-FT-ICR MS. The results indicate important anti-H. pylori, antioxidant and immunomodulatory activities from Eugenia uniflora highlighting its importance in the prevention and treatment of diseases caused by H. pylori infection.

Evaluation of antimicrobial, hypotensive and diuretic effect of Eugenia uniflora extracts / Avaliação do efeito antimicrobiano, hipotensivo e diurético de extratos de Eugenia uniflora

The genus Eugenia sp. (Myrtaceae) comprises plants with reported antioxidant and antidiarrheal capability among other therapeutic potentials. The aim of this study was to evaluate the antimicrobial activity of essential oil; diuretic and hypotensive activities of aqueous extracts from leaves of Eugenia uniflora. The antimicrobial activity was evaluated . The diuretic and hypotensive activities were evaluated in normotensive Wistar rats by measuring blood pressure and urine flow after received four different concentrations of aqueous extracts (10%, 15%, 20% and 25%). Essential oil inhibited the growth of Candida parapsilosis and Candida albicans with MIC values lower than 14.41 mg/mL, equal to 57.75 mg/mL for Candida krusei. Among antibacterial effect, essential oil inhibited growth with a MIC equals to 153.93 mg/mL for all strains tested, except for Escherichia coli (MIC equals to 307.96 mg/mL. Aqueous extracts showed powerful reductions of the arterial pressure (34% and 31% lower than the control), after administration of 10% and 25% of aqueous extract, respectively. However, the animals that received the aqueous extract at the 15% and 20% concentrations presented a discrete hypotensive effect (20% and 21% lower than control group, respectively) concomitantly to powerful diuretic effect (280% and 91% higher than control group, respectively). These data confirmed the potential biological effect of this species, and represents an important step toward a depth study on the therapeutic properties of this species

O gênero Eugenia sp. (Myrtaceae) compreende plantas com capacidade antimicrobiana e antioxidante entre outros potenciais terapêuticos. O objetivo deste estudo foi avaliar a atividade antimicrobiana de óleo essencial; atividade diurética e hipotensora de extrato aquoso de folhas de Eugenia uniflora. A atividade antimicrobiana foi avaliada pela determinação da concentração inibitória mínima (MIC) e concentração mínima bactericida (MBC) de cepas bacterianas e concentração fungicida mínima (MFC) para fungos. A atividade diurética e hipotensora foi avaliada em ratos Wistar normotensos pela mensuração da pressão sanguínea e fluxo urinário após administração de quatro diferentes concentrações de extrato aquoso (10%, 15%, 20% e 25%). Óleo essencial inibiu o crescimento de Candida parapsilosis e Candida albicans com valores de MIC menores que 14,41 mg/mL, igual a 57,75 mg/mL para Candida krusei. A respeito do efeito antimicrobiano, o óleo essencial inibiu o crescimento de todas as cepas testadas, com MIC igual a 153,93 mg/mL, exceto para Escherichia coli (MIC igual a 307.96 mg/mL). O extrato aquoso mostrou redução importante da pressão arterial (34% e 31% quando comparado ao controle), após administração de 10% e 25% do extrato aquoso, respectivamente. Contudo, os animais que receberam o extrato aquoso na concentração de 15% e 20% apresentaram discreto efeito hipotensor (20% e 21% menor que o grupo controle, respectivamente) concomitantemente ao importante efeito diurético (280% e 91% maior quando comparado ao grupo controle, respectivamente). Esses achados confirmam o potencial efeito biológico dessa espécie, e representa um importante embasamento para estudos relacionados as propriedades terapêuticas da Eugenia uniflora

Chromatographic profiles of extractives from leaves of Eugenia uniflora

ABSTRACT Eugenia uniflora L., Myrtaceae, popularly known as "pitanga", is used in traditional medicine due the properties attributed to its chemical content, these being mainly hydrolysable tannins and flavonoids. This study provides a qualitative and quantitative evaluation of chemical profile from leaves of E. uniflora. The HPLC analysis was carried out on a C18 column (4.6 mm × 250 mm, 5 µm) by gradient elution with methanol and water (acidified with trifluoracetic acid); and silica gel Plates 60-F 254 with 10-12 µm and 5-6 µm particles, respectively for TLC and High HPTLC analysis. The chromatographic data obtained from HPLC, TLC and HPTLC presented bands and peaks related to flavonoids (myricitrin and derivatives) and tannins (gallic and ellagic acids), which were observed from different samples. The chromatographic similarities enabled the building of a typical fingerprint for the herbal material. The similarity analysis of the sample data by Pearson correlation showed R values >0.9 among peaks (HPLC) and bands (HPTLC). In addition, the analytical methodology developed by HPLC enabled the satisfactory quantification of marker substances [ellagic acid = 0.22% and 0.20% (m/m); gallic acid = 0.20% and 0.43%; myricitrin = 0.42 and 1.74% (m/m) in herbal drug and crude extract, respectively]. The procedure was also validated in accordance with the assays required by Brazilian legislation. Thus, the HPTLC and HPLC methods developed in this study provide helpful and simple tools for the quality evaluation both qualitatively and quantitatively of raw materials and extractives from leaves of E. uniflora.

Antineoplastic potential of the aqueous crude extract of Eugenia uniflora L. in human cervical cancer

Abstract Considering the high prevalence of human cervical cancer and the adverse effects of the available treatments, it is important to develop studies involving plants. Eugenia uniflora L. is a Brazilian native plant widely used in folk medicine and some biological effects have already been described. In this study, we investigated the biologicals effects of the aqueous crude extract of E. uniflora leaves in relation to the viability of human cervical cancer cells (SiHa), non-tumorigenic cells HaCaT and human lymphocytes. Our results demonstrated that different concentrations of E. uniflora's extract significantly inhibited the viability of the Siha cell line at 24, 48 and 72 hours of treatment, but did not induce significant changes in the HaCat cell line and human lymphocytes. Tumor cells had adhesion capacity, migration processes, ability of colony forming and the potential to recover its viability after treatment. withdrawal, significantly reduced. The nuclear morphology revealed chromatin condensation, and the flow cytometry showed predominantly cell death by apoptosis in the treated tumor cells. Therefore, the E. uniflora's extract may contribute for future studies aiming at new therapeutic perspectives for human cervical cancer.

LC-MS characterization, anti-kinetoplastide and cytotoxic activities of natural products from Eugenia jambolana Lam.and Eugenia uniflora

Objective:To evaluate the trypanocidal,leishmanicidal and cytotoxic activity of Eugenia jambolana (E.jambolana) and Eugenia uniflora (E.uniflora) extracts and fractions.Methods:The products were characterized by LC-MS.Antiparasitic assays were performed and cytotoxicity was evaluated in fibroblastos.In vitro assays were performed using spectrophotometric evaluation.All assays were performed in thrice.Results:The results showed that the extracts and the tannic fraction from E.jambolana inhibited 100％ of the epimastigote lines.The ethanolic extract was the most efficient in all concentrations tested against the three parasite strains.In the cytotoxicity assay the flavonoid fraction showed low toxicity.All E.uniflora samples showed cytotoxicity at the highest concentration tested,but the extract showed no toxic effect on the fibroblasts at the lowest concentration.The flavonoid and tannic fractions were more efficient against Leishmania braziliensis promastigotes compared to the extract.However,the extracts and the tannic fraction were more effective against Leishmania infantum strains.The effect on epimastigote cells was observed at all concentrations tested,with all E.uniflora samples.However,the samples were more effective at the highest concentration,where there was inhibition in 100％ of the Trypanosoma cruzi strains.Contusions:The species E.jambolana and E.uniflora presented antiparasitic activity against all tested parasite strains,indicating that these species can serve as an altemative therapy as they were efficient in the tests performed.The E.uniflora extract and the E.jambolana flavonoid fraction presented a low cytotoxicity,opening the floor for new biological studies.

Antioxidant activity, antibacterial and inhibitory effect of intestinal disaccharidases of extracts obtained from Eugenia uniflora L. Seeds

Abstract The use of medicinal plants for disease prevention, treatment and cure is an ancient practice used by humanity, and many plants species are used in bioprospecting research. In this context, its stands out Eugenia uniflora L., populary known as pitangueira and belongs to the Myrtaceae family, with a wide geographic distribution and native of Brazil. In view of the therapeutic qualities of the plant and the lack of the studies on its seeds, the present study had as objective to evaluate the phytochemical profile of the extracts of Eugenia uniflora L. seeds, from different solvents, as well as their antibacterial activity, antioxidant and its inhibitory effect of intestinal disaccharidases. Results showed a high content of phenolic compounds and total flavonoids, thus characterizing antioxidant activity, also highlighting the best bacteriostatic action for the Gram positive strain of Staphylococcus aureus in the ethanolic fraction. Regarding the disaccharidases, a strong inhibitory action was observed for all concentrations, evidencing a antihyperglycemic potential. The present research allowed to concluded that Eugenia uniflora L. seeds have promising biological activities for the industrial sector, but a more detailed investigation is needed regarding their bioactive compounds.

Resumo A utilização de plantas com fins medicinais para prevenção, tratamento e cura de doenças é uma prática antiga utilizada pela humanidade, sendo que muitas espécies vegetais são usadas para a pesquisa da bioprospecção. Neste contexto, destaca-se a Eugenia uniflora L., conhecida popularmente como pitangueira e pertencente à família Myrtaceae, com ampla distribuição geográfica e nativa do Brasil. Diante das qualidades terapêuticas da planta e a falta de estudos sobre suas sementes, o presente trabalho teve como objetivo avaliar o perfil fitoquímico dos extratos das sementes de Eugenia uniflora L. a partir de diferentes solventes, bem como sua atividade antibacteriana, antioxidante e seu efeito inibidor de dissacaridases intestinais. Os resultados mostraram um alto teor de compostos fenólicos e flavonóides totais, caracterizando a atividade antioxidante, destacando também a melhor ação bacteriostática para a cepa Gram positiva de Staphylococcus aureus na fração etanólica. Em relação às dissacaridases, uma forte ação inibitória foi observada para todas as concentrações, evidenciando um potencial anti-hiperglicêmico. A presente pesquisa permitiu concluir que as sementes de Eugenia uniflora L. apresentam atividades biológicas promissoras para o setor industrial, mas é necessária uma investigação mais detalhada de seus compostos bioativos.

LC–MS characterization, anti-kinetoplastide and cytotoxic activities of natural products from Eugenia jambolana Lam. and Eugenia uniflora

Objective To evaluate the trypanocidal, leishmanicidal and cytotoxic activity of Eugenia jambolana (E. jambolana) and Eugenia uniflora (E. uniflora) extracts and fractions. Methods The products were characterized by LC–MS. Antiparasitic assays were performed and cytotoxicity was evaluated in fibroblastos. In vitro assays were performed using spectrophotometric evaluation. All assays were performed in thrice. Results The results showed that the extracts and the tannic fraction from E. jambolana inhibited 100% of the epimastigote lines. The ethanolic extract was the most efficient in all concentrations tested against the three parasite strains. In the cytotoxicity assay the flavonoid fraction showed low toxicity. All E. uniflora samples showed cytotoxicity at the highest concentration tested, but the extract showed no toxic effect on the fibroblasts at the lowest concentration. The flavonoid and tannic fractions were more efficient against Leishmania braziliensis promastigotes compared to the extract. However, the extracts and the tannic fraction were more effective against Leishmania infantum strains. The effect on epimastigote cells was observed at all concentrations tested, with all E. uniflora samples. However, the samples were more effective at the highest concentration, where there was inhibition in 100% of the Trypanosoma cruzi strains. Conclusions The species E. jambolana and E. uniflora presented antiparasitic activity against all tested parasite strains, indicating that these species can serve as an alternative therapy as they were efficient in the tests performed. The E. uniflora extract and the E. jambolana flavonoid fraction presented a low cytotoxicity, opening the floor for new biological studies.

Eugenia uniflora Dentifrice for Treating Gingivitis in Children: Antibacterial Assay and Randomized Clinical Trial

Abstract School-age children are frequently at high risk for the onset of biofilm-dependent conditions, including dental caries and periodontal diseases. The objective of this study was to evaluate the clinical efficacy of a dentifrice containing Eugenia uniflora Linn. (Surinam cherry) extract versus a triclosan-based comparator in treating gingivitis in children aged 10-12 years. The in vitro antibacterial potential of the dentifrice was tested against oral pathogens (Streptococcus mutans, Streptococcus oralis and Lactobacillus casei). Then a phase-II clinical trial was conducted with 50 subjects aged 10-12 years, with clinical signs of gingivitis. The subjects were randomly assigned to the experimental group (n=25) and control group (n=25), in which participants used the experimental dentifrice and a triclosan-based fluoridated dentifrice (Colgate Total 12(r)), respectively. Clinical examinations assessed the presence of gingivitis (primary outcome) and biofilm accumulation (secondary outcome) using the Gingival-Bleeding Index (GBI) and Simplified Oral Hygiene Index (OHI-S), respectively, at baseline and after seven days of tooth brushing 3x/day. The data were analyzed using paired and unpaired t-test (GBI) and Wilcoxon and Mann-Whitney (OHI-S), with p≤0.05. The experimental dentifrice showed efficient antibacterial activity in vitro. In the clinical trial, a significant reduction in gingival bleeding was observed in both experimental and control groups (p<0.0001), with no statistical difference between them (p=0.178), although a small size effect was observed. Biofilm accumulation was only reduced in the control group (p=0.0039). In conclusion, E. uniflora dentifrice showed anti-gingivitis properties in children aged 10-12 years. Thus, it may be a potentially efficient and safe product to be used alternatively in preventive dental practice.

Resumo Crianças em idade escolar apresentam, com frequência, alto risco para o desenvolvimento de doenças biofilme-dependentes, incluindo cárie e doenças periodontais. Este estudo investigou a eficácia clínica de um dentifrício contendo o extrato de Eugenia uniflora Linn. (pitanga) comparado a um dentifrício com triclosan no combate à gengivite em crianças de 10 a 12 anos. Foi avaliado o potencial antibacteriano in vitro do dentifrício sobre microorganismos da cavidade oral (S. mutans S. oralis and L. casei) e realizado um estudo clínico fase II incluindo 50 sujeitos, com sinais clínicos de gengivite, divididos aleatoriamente em dois grupos: Experimental - 25 sujeitos usaram o dentifrício contendo extrato de pitanga; e Controle - 25 sujeitos que usaram um dentifrício fluoretado contendo triclosan (Colgate Total 12(r)). Na baseline e após sete dias consecutivos de escovação, foi realizado o exame clínico para diagnóstico de gengivite (desfecho primário) e acúmulo de biofilme (desfecho secundário), utilizando o Índice de Sangramento Gengival (ISG) e Índice de Higiene Oral Simplificado (IHO-S). Os dados foram analisados utilizando o teste t pareado e não pareado (ISG) e teste de Wilcoxon e Mann-Whitney (IHO-S), com nível de significância de p≤0,05. Observou-se uma efetiva atividade antibacteriana do dentifrício experimental. No estudo clínico, observou-se redução de sangramento gengival em ambos os grupos experimental e controle (p<0,0001), não havendo diferença entre eles (p 0,178), embora com uma pequena magnitude de efeito. Apenas o grupo controle reduziu significantemente o acúmulo de biofilme (p=0,0039). Concluiu-se que o dentifrício experimental de E. uniflora mostrou-se eficaz na redução de gengivite em crianças de 10 a 12 anos. Assim, este dentifrício apresenta potencial para ser utilizado de forma eficaz e segura em odontologia preventiva.

Ação dos extratos de quatro plantas sobre larvas infectantes de nematódeos gastrintestinais de ovinos / Acción de extractos de cuatro plantas en las larvas infectivas de nematodos gastrointestinales de ovinos / Action of extracts of four plants against infective larvae of gastrointestinal nematodes of sheep

Introdução: a ação de extratos hidroalcoólicos de Eugenia uniflora L. (pitangueira), Mentha x piperita L. (hortelã), Myrcianthes pungens (O. Berg) D. Legrand (guabiju) e Peltophorum dubium (Spreng.) Taub. (canafístula) foi avaliada sobre o desenvolvimento de nematódeos gastrintestinais nas coproculturas de ovinos. Objetivo: avaliar a ação in vitro dos extratos vegetais sobre os nematódeos gastrintestinais de ovinos. Métodos: cada extrato foi testado em culturas triplicadas de fezes nas seguintes concentrações: 200, 100, 20 e 2 mg.mL-1. Como controle positivo foi utilizado moxidectina (0,001 mg.mL-1) e água destilada como controle negativo. Após incubação, as larvas recuperadas foram fixadas, coradas e conservadas para posterior leitura. Resultados: a concentração de 200 mg.mL-1 do extrato de Eugenia uniflora foi a que promoveu menor número de larvas infectantes recuperadas das coproculturas de ovinos (número de larvas por grama de fezes= 19) quando comparada as outras concentrações. Este valor foi dez vezes menor que o ldpg recuperado das coproculturas tratadas com o mesmo extrato na menor concentração (2 mg.mL-1). Conclusão: a ação do extrato de Eugenia uniflora na maior concentração não diferiu estatisticamente do controle positivo (p< 0,05), sugerindo um efeito sobre a cultura destes nematódeos gastrointestinais.

Introducción: la acción de los extractos hidroalcohólicos de Eugenia uniflora L. (pitangueira), Mentha x piperita (menta), Myrcianthes pungens (O. Berg) D. Legrand (guabiju) y Peltophorum dubium (Spreng.) Taub. (cañafístula) se valoró en el desarrollo de los nematodos gastrointestinales en cultivos fecales de ovinos. Objetivo: evaluar el efecto in vitro de extractos de plantas sobre los nematodos gastrointestinales de ovinos. Métodos: cada extracto se probó en cultivos por triplicado de las heces en las concentraciones siguientes: 200 mg.mL-1, 100 mg.mL-1, 20 mg.mL-1 y 2 mg.mL-1. Se utilizó como control positivo moxidectina (0,001 mg.mL-1) y agua destilada como control negativo. Después de la incubación, las larvas se recuperaron, se fijaron y tiñeron, y se conservaron para su posterior lectura. Resultados: la concentración de 200 mg.mL-1 de extracto de Eugenia uniflora se promovió a la disminución del número de larvas infectantes, se recuperó a partir de cultivos de heces de ovejas (número de larvas por gramos de heces= 19) en comparación con otras concentraciones. Este valor fue 10 veces menor que el recuperado de coprocultivos tratados con la concentración más baja en el extracto crudo (2 mg.mL-1). Conclusión: la acción del extracto de Eugenia uniflora en la mayor concentración no difirió estadísticamente del control positivo (p< 0,05), lo que sugiere un efecto en el cultivo de los nematodos gastrointestinales.

Introduction: an assessment was made of the action of hydroalcoholic extracts of Eugenia uniflora L. (pitangueira), Mentha x piperita (menta), Myrcianthes pungens (O. Berg) D. Legrand (guabiju) and Peltophorum dubium (Spreng.) Taub. (cañafistula) on the development of gastrointestinal nematodes in fecal cultures from sheep. Objective: evaluate the in vitro effect of the plants upon gastrointestinal nematodes of sheep. Methods: Each extract was tested in triplicate cultures of the faeces at the following concentrations: 200 mg.mL-1, 100 mg.mL-1, 20 mg.mL-1 and 2 mg.mL-1. moxidectin (0.001 mg.mL-1) was used as positive control, and distilled water as negative control. After incubation, the larvae were recovered, fixed and stained, and preserved for future reading. Results: the 200 mg.mL-1 concentration of Eugenia uniflora extract was promoted to a decrease in the number of infective larvae, and was recovered from cultures of sheep faeces (number of larvae per gram of faeces = 19), in comparison with other concentrations. This value is 10 times smaller than the one recovered from coprocultures treated with the lowest concentration in the crude extract (2 mg.mL-1). Conclusion: statistically, the action of the extract of Eugenia uniflora at the highest concentration did not differ from the positive control (p< 0.05). This is suggestive of an effect on the culture of gastrointestinal nematodes.

Atividade leishmanicida in vitro de Eugenia uniflora e Momordica charantia / Antileishmanial in vitro activity of Eugenia uniflora and Momordica charantia

A Leishmaniose Tegumentar Americana no Brasil é causada por uma variedade de espécies de Leishmania e uma grande diversidade destes parasitas pode ser encontrada na Região Amazônica. Revisões recentes na quimioterapia de leishmaniose enfatizam as deficiências dos agentes terapêuticos atualmente disponíveis e mostram a necessidade urgente de novos candidatos. Uma alternativa para substituir esses medicamentos são extratos naturais de Eugenia uniflora e Momordica charantia. Foram preparados extratos etanólicos das folhas de E. uniflora e M. charantia. Para os testes in vitro de Leishmania brasiliensis foram utilizadas formas promastigotas. O ensaio de citotoxicidade foi realizado com linhagens de fibroblastos. Nossos resultados indicam que E. uniflora foi eficaz contra a cepa de parasita testada, representando uma fonte alternativa de produtos naturais com atividade contra L. brasiliensis.

Cutaneous leishmaniasis is caused in Brazil by several species of the genus Leishmania and a wide variety of these protozoan parasites can be found in Brazil, mainly in the Amazon region. Recent reviews on the chemotherapy of leishmaniasis show the low effectiveness of the usual therapeutic agents, demonstrating the need for new drugs. An interesting possible alternative to the conventional drugs is offered by natural products extracted from Eugenia uniflora and Momordica charantia. Ethanol extracts were prepared from the leaves of Eugenia uniflora and Momordica charantia and assayed in vitro against Leishmania brasiliensis promastigotes and fibroblasts to assess their antileishmanial and cytotoxic activities, respectively. Our results indicate that E. uniflora was active against the parasitic forms of L. brasiliensis

Pré-tratamento com água e doses de ácido indolbutírico para estaquia herbácea de pitangueiras / Pre-treatments with water and indole butyric acid dosis for herbaceous cuttings of Surinam cherry

No Brasil, a maioria dos pomares de pitangueira (Eugenia uniflora) é formada por mudas do tipo pé-franco, o que torna os plantios com baixa uniformidade genética. A propagação vegetativa via estaquia é uma alternativa viável para propagação de diversas espécies frutíferas, podendo ser utilizada também com as espécies nativas, proporcionando a formação de pomares homogêneos, com a produção de mudas com alta sanidade, além de antecipar a produção. Nesse contexto, conduziu-se um estudo visando à multiplicação vegetativa da pitangueira, em que se testou a estaquia herbácea a partir de estacas coletadas de plantas matrizes jovens e adultas, submetidas a três períodos de imersão em água (0, 24 ou 48 horas), além da aplicação de doses de ácido indolbutírico (0, 2000, 4000 e 6000mg L-1). Foram avaliadas a sobrevivência ( por cento), a retenção e a emissão foliar (n° folhas/ estaca), a calogênese ( por cento) e enraizamento das estacas ( por cento). Ao final do experimento, avaliou-se massa fresca e seca de parte aérea e raiz, número de folhas e área foliar. O delineamento experimental foi o completamente casualizado, com três repetições de dez plantas por tratamento para os parâmetros não destrutivos e três repetições de cinco plantas por tratamento para os parâmetros destrutivos. A estaquia herbácea foi eficiente para produzir mudas de pitangueira, especialmente quando utilizadas estacas oriundas de plantas jovens sem necessidade de pré-lavagem nem adição de auxinas exógenas.

In Brazil, most of the Surinam cherry (Eugenia uniflora) orchards is formed by ungrafted plants, which makes the plantations have low genetic uniformity. The vegetative minicutting propagation is a viable alternative for the propagation of several fruit species, and can also be used with native species providing the formation of orchards with homogeneous populations of plants, with production of seedlings with high health and accelerated propagation process. In this context, a study for the multiplication of Surinam cherry was conduced with herbaceous cuttings, taken from young and adult mother plants, immersed in indole butyric acid doses (0, 2000, 4000 and 6000mg L-1), in three different water periods (0, 24 or 48 hours). Survival rate ( percent), leaf retention and emission (No. leaves /cutting), callus rate ( percent) and rooting ( percent) were evaluated. At the end of the experiment, were also evaluated fresh and dry weight of shoot and root, leaf number and leaf area. The experiment has a completely randomized design with three replications of ten plants per treatment for nom destructive and three replications of five plants per treatment for the destrutive parameters. The herbaceous cuttings were efficient to produce seedlings of Surinam cherry, particularly when using cuttings derived from seedlings in the absence of the immersion in water and exogenous auxin.

Influence of foliar nutrients on phenol levels in leaves of Eugenia uniflora

Eugenia uniflora L., Myrtaceae, leaves contain high amounts of phenolic compounds which are responsible for several pharmacological activities. In order to evaluate the phenolics seasonal variation leaves were analysed on a monthly basis during the period of two years for the contents of hydrolysable tannins, total phenols, flavonoids, and nutrients (N, P, K, S, Ca, Mg, Mn, Zn, Cu, and Fe). Results were correlated with climate conditions (rainfall, humidity, and mean temperature) by Principal Component and ClusterAnalysis which allowed four groups to be distinguished with respect to the age of the leaves and the content of some metals. Young leaves were characterised by high levels of Zn and nitrogen whereas old leaves contained high levels of Fe and calcium, and both groups had moderate amounts of phenolics. Adult leaves were divided in two groups and results revealed that while one group had the highest levels of all phenols and lowest amounts of Mn and Cu, the other showed opposite quantities. The Canonical Correlation Analysis confirmed a highly significant negative correlation between phenol contents and Mn and Cu. These facts suggested that flavonoids and tannins production depends of the amounts of foliar nutrients, Cu and Mn in particular, which are cofactors of enzymes involved in phenol degradation and lignin biosynthesis. This knowledge can improve this specie cultivation in order to enhance the phenolic compounds concentration.

A new approach for quantifying furanodiene and curzerene: a case study on the essential oils of Eugenia uniflora L., Myrtaceae (pitangueira) leaves

The essential oil obtained from the leaves of Eugenia uniflora L., Myrtaceae, which grows in the Brazilian savannah, was studied by gas chromatography mass spectrometry (GC-MS). Furanodiene (1.2 percent) was thermally rearranged to curzerene (85.1 percent) to produce a combined content of 86.3 percent. GC analysis carried out under mild conditions (with a constant temperature of 100 ºC) showed that the furanodiene concentration was three-fold greater than the curzerene concentration, i.e., the essential oil contained 64.7 percent furanodiene and 21.6 percent curzerene. Germacrene B also rearranged to γ-elemene and the concentration of both was 2.3 percent. Special care should be taken when conventional gas chromatography analysis is used for quantifying compounds that can rearrange at high temperatures.

Seasonal variation in the phenol content of Eugenia uniflora L. leaves / Variação sazonal nos teores de fenóis de folhas de Eugenia uniflora L

Hydrolysable tannins, total phenols and flavonoids in Eugenia uniflora leaves were monthly analysed for one year. The results were correlated with climate conditions (rainfall, humidity, cloudiness and mean temperature) through chemometric methods. Principal component analysis revealed high levels of hydrolysable tannins in the rainy season, whereas flavonoids were mainly produced in the dry season. These facts suggest that climatic changes may be one of the factors affecting phenol levels in Eugenia uniflora.

Taninos hidrolisáveis, fenóis totais e flavonóides presentes em folhas de Eugenia uniflora foram quantificados mensalmente durante um ano. Os resultados foram correlacionados com as condições climáticas (pluviosidade, umidade, nebulosidade e temperatura média) através de métodos quimiométricos. Análise de componentes principais revelou a ocorrência de altos teores de taninos hidrolisáveis durante a estação de chuvas, enquanto os flavonóides foram produzidos principalmente na estação seca. Estes fatos sugerem que mudanças climáticas podem ser um dos fatores que afetam os níveis de fenóis em Eugenia uniflora.

Repetibilidade de caracteres de fruto em araçazeiro e pitangueira / Repeatability traits of strawberry guava and surinam cherry fruits

O objetivo deste trabalho foi estimar a repetibilidade de carateres de fruto em araçazeiro e pitangueira. Foram avaliados, em quatro ciclos produtivos, os caracteres peso, diâmetro, sólidos solúveis totais e produção de frutos em 10 seleções de araçazeiro e peso, diâmetro e sólidos solúveis totais em nove seleções de pitangueira, do Banco de Germoplasma da Embrapa Clima Temperado, em Pelotas-RS. O coeficiente de repetibilidade, o número de medições necessárias e o coeficiente de determinação foram estimados pelos métodos de análise de variância, componentes principais e análise estrutural. O coeficiente de repetibilidade foi mais eficientemente estimado pelo método dos componentes principais, com base na matriz de covariâncias. O caractere peso de fruto é mais adequado para ser utilizado na seleção fenotípica em araçazeiro e pitangueira. É necessária apenas uma medição para predizer o valor de peso de fruto de genótipos de araçazeiro, com 90 por cento de acurácia. Com quatro anos de avaliação, é possível inferir o valor real com confiabilidade acima de 80 por cento para todos os caracteres. Isso facilita a seleção fenotípica para esses caracteres em programas de melhoramento de araçazeiro e pitangueira.

The aim of this research was to estimate the repeatability traits of strawberry guava and surinam cherry fruits. It was evaluated in four productive cycles, the traits weight, diameter, total soluble solids and fruit production in ten selection of strawberry guava and weight, diameter and total soluble solids in nine selection of surinam cherry from the Germplasm Bank of the Embrapa Clima Temperado, in Pelotas-RS, Brazil. The repeatability coefficient, minimum observations number and determination coefficient were estimated by analysis of variance, principal components analysis and structural analysis. The repeatability coefficient was better estimated by the method of principal components analysis, based on covariance matrix. The fruit weight is adequate to phenotypic selection in strawberry guava and surinam cherry. It's only required a measurement to obtain the value of fruit weight value of strawberry guava selections, with 90 percent accuracy. With four years of evaluation, it's possible to infer the actual value with reliability above 80 percent for all characters. This facilitates phenotypic selection for these traits in strawberry guava and surinam cherry breeding programs.