Chlorophyll treatment combined with photostimulation increases glycolysis and decreases oxidative stress in the liver of type 1 diabetic rats

Photodynamic therapy (PDT) promotes cell death, and it has been successfully employed as a treatment resource for neuropathic complications of diabetes mellitus (T1DM) and hepatocellular carcinoma. The liver is the major organ involved in the regulation of energy homeostasis, and in pathological conditions such as T1DM, changes in liver metabolic pathways result in hyperglycemia, which is associated with multiple organic dysfunctions. In this context, it has been suggested that chlorophyll-a and its derivatives have anti-diabetic actions, such as reducing hyperglycemia, hyperinsulinemia, and hypertriglyceridemia, but these effects have not yet been proven. Thus, the biological action of PDT with chlorophyll-a on hepatic parameters related to energy metabolism and oxidative stress in T1DM Wistar rats was investigated. Evaluation of the acute effects of this pigment was performed by incubation of isolated hepatocytes with chlorophyll-a and the chronic effects were evaluated by oral treatment with chlorophyll-based extract, with post-analysis of the intact liver by in situ perfusion. In both experimental protocols, chlorophyll-a decreased hepatic glucose release and glycogenolysis rate and stimulated the glycolytic pathway in DM/PDT. In addition, there was a reduction in hepatic oxidative stress, noticeable by decreased lipoperoxidation, reactive oxygen species, and carbonylated proteins in livers of chlorophyll-treated T1DM rats. These are indicators of the potential capacity of chlorophyll-a in improving the status of the diabetic liver.

Effect of inactive dry yeast from sugar cane as protein source on rumen fermentation in Saanen goats / Efeito da levedura seca inativa da cana-de-açúcar como fonte de proteína sobre a fermentação ruminal de caprinos Saanen

Five castrated and ruminally cannulated Saanen goats (±48.19kg) were used to evaluate intake, digestibility and rumen fermentation parameters of diets with inactive dry yeast as a soybean meal substitute (0, 25, 50, 75 and 100 percent). Goats were randomly assigned to a 5×5 (five levels of dry yeast x five periods) Latin square design. Diets were composed of corn silage (40 percent), ground corn, soybean meal and/or dry yeast and mineral supplement. The intake and digestibility of DM, OM, CP, NDF and TC were not influenced by the treatments. However, EE intake showed negative linear effect. The TDN content did not change with the inclusion of dry yeast in the diets. The pH, N-NH3 concentration and rumen short-chain fatty acids content did not differ among diets. Rumen content scanning electron microscopy observations did not suggest microbial colonization and degradation changes. Dry yeast from sugar cane can replace soybean meal in diets for Saanen goats without changing the rumen fermentation pattern, intake and digestibility.

Com o objetivo de avaliar a ingestão, a digestibilidade e os parâmetros de fermentação ruminal de rações com de levedura seca em substituição ao farelo de soja (0; 25; 50; 75 e 100 por cento), foram utilizados cinco caprinos Saanen (± 48,19kg) castrados e canulados no rúmen distribuídos em quadrado latino 5 x 5 (cinco níveis x cinco períodos). As rações foram compostas de silagem de milho (40 por cento), milho moído, farelo de soja e/ou levedura seca e suplemento vitamínico-mineral. A ingestão e a digestibilidade da MS, MO, PB, FDN e dos CT não foram alteradas. A ingestão de EE apresentou efeito linear negativo. O NDT não foi alterado com a inclusão da levedura seca. O pH e as concentrações de N-NH3 e de ácidos graxos de cadeia curta no rúmen não apresentaram alterações. As observações por microscopia eletrônica de varredura não indicaram mudanças na colonização microbiana e na degradação da digesta ruminal. A levedura seca da cana-de-açúcar não altera o padrão de fermentação ruminal, a ingestão e a digestibilidade, podendo substituir o farelo de soja em rações de caprinos Saanen.

Effects of a methanolic fraction of soybean seeds on the transcriptional activity of peroxisome proliferator-activated receptors (PPAR)

Since the anti-inflammatory, antidiabetic and hypolipidemic effects of soy isoflavones may be mediated by activation of peroxisome proliferator-activated receptors (PPAR), the present study investigated whether the methanolic fractions obtained from soybean seeds (E1) and soybean seed coats with hypocotyls (E2) could influence PPARα, PPARγ and PPARβ/δ transcriptional activity. The isoflavones from E1 and E2 were quantified by HPLC analysis. E1 and E2 were rich in isoflavones (daidzin, glycitin, genistin, malonyldaidzin, malonylglycitin, malonylgenistin, daidzein, glycitein, and genistein). Moreover, E1 and E2 showed no evidence of genetically modified material containing the gene CP4 EPSPS. To investigate PPAR transcriptional activity, human promonocytic U-937 cells were treated with E1 and E2 (200, 400, 800, and 1600 µg/mL), positive controls or vehicle. Data are reported as fold-activation of the luciferase reporter driven by the PPAR-responsive element. Dose-response analysis revealed that E1 and E2 induced the transcriptional activity of PPARα (P < 0.001), with activation comparable to that obtained with 0.1 mM bezafibrate (positive control) at 1600 µg/mL (4-fold) and 800 µg/mL (9-fold), respectively. In addition, dose-response analysis revealed that E1 and E2 activated PPARβ/δ (P < 0.05), and the activation at 800 µg/mL (4- and 9-fold, respectively) was comparable to that of 0.1 mM bezafibrate (positive control). However, no effect on PPARγ was observed. Activation of PPARα is consistent with the lipid-lowering activity of soy isoflavones in vivo, but further studies are needed to determine the physiological significance of PPARβ/δ activation.

Antibacterial activity of indole alkaloids from Aspidosperma ramiflorum

We evaluated the antibacterial activities of the crude methanol extract, fractions (I-V) obtained after acid-base extraction and pure compounds from the stem bark of Aspidosperma ramiflorum. The minimum inhibitory concentration (MIC) was determined by the microdilution technique in Mueller-Hinton broth. Inoculates were prepared in this medium from 24-h broth cultures of bacteria (10(7) CFU/mL). Microtiter plates were incubated at 37°C and the MICs were recorded after 24 h of incubation. Two susceptibility endpoints were recorded for each isolate. The crude methanol extract presented moderate activity against the Gram-positive bacteria B. subtilis (MIC = 250 µg/mL) and S. aureus (MIC = 500 µg/mL), and was inactive against the Gram-negative bacteria E. coli and P. aeruginosa (MIC > 1000 µg/mL). Fractions I and II were inactive against standard strains at concentrations of <=1000 µg/mL and fraction III displayed moderate antibacterial activity against B. subtilis (MIC = 500 µg/mL) and S. aureus (MIC = 250 µg/mL). Fraction IV showed high activity against B. subtilis and S. aureus (MIC = 15.6 µg/mL) and moderate activity against E. coli and P. aeruginosa (MIC = 250 µg/mL). Fraction V presented high activity against B. subtilis (MIC = 15.6 µg/mL) and S. aureus (MIC = 31.3 µg/mL) and was inactive against Gram-negative bacteria (MIC > 1000 µg/mL). Fractions III, IV and V were then submitted to bioassay-guided fractionation by silica gel column chromatography, yielding individual purified ramiflorines A and B. Both ramiflorines showed significant activity against S. aureus (MIC = 25 µg/mL) and E. faecalis (MIC = 50 µg/mL), with EC50 of 8 and 2.5 µg/mL for ramiflorines A and B, respectively, against S. aureus. These results are promising, showing that these compounds are biologically active against Gram-positive bacteria.

Antiprotozoal and molluscicidal activities of five Brazilian plants

Leishmaniasis, Chagas' disease and schistosomiasis (bilharzia) are parasitic diseases with wide distribution on the American continent, affecting millions of people. In the present study, biological assays for antiprotozoal and molluscicidal activities were carried out with ethanolic extracts of plant species from the Brazilian part of the Upper Paraná River. Crude extracts were obtained by percolation with absolute ethanol from the leaves of Cayaponia podantha Cogn., Nectandra falcifolia (Nees) Castiglioni and Paullinia elegans Cambess., as well as from the aerial parts of Helicteres gardneriana St. Hil. & Naud. and Melochia arenosa Benth., all belonging to genera used in folk medicine. Trypanocidal activity of plants was assayed on epimastigote cultures in liver infusion tryptose. Anti-leishmanial activity was determined over cultures of promastigote forms of the parasite in Schneider's Drosophila medium. Microscopic countings of parasites, after their incubation in the presence of different concentrations of the crude extracts, were made in order to determine the percentage of growth inhibition. C. podantha and M. arenosa, at a concentration of 10 æg/mL, showed 90.4 ± 11.52 and 88.9 ± 2.20 percent growth inhibition, respectively, of epimastigote forms of Trypanosoma cruzi, whereas N. falcifolia demonstrated an LD50 of 138.5 æg/mL against promastigote forms of Leishmania (Viannia) braziliensis. Regarding molluscicidal activity, the acute toxicity of the extracts on Biomphalaria glabrata was evaluated by a rapid screening procedure. M. arenosa was 100 percent lethal to snails at 200 æg/mL and showed an LD50 of 143 æg/mL. Screening of plant extracts represents a continuous effort to find new antiparasitic drugs.

Neolignanas e análise do óleo essencial das folhas de Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck / Neolignans and the analysis of the essential oil of Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck leaves

O óleo essencial das folhas de Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck Piparaceae, coletadas no Horto de Plantas Medicinais da Universidade Estadual de Maringá, foi obtido por hidrodestilação. Uma análise preliminar por CG/EM e RMN 13C foi realizada. O b-mirceno (70 por cento) foi identificado como componente majoritário através da comparação dos espectros de massa e RMN 13C com dados da literatura. Quatro neolignanas foram isoladas do extrato hidroetanólico das folhas e identificadas: eupomatenóide-6, eupomatenóide-5, eupomatenóide-3 e conocarpano. As estruturas dessas substâncias foram estabelecidas por meio de estudos de RMN ¹H e 13C, ¹H x ¹H - COSY, HETCOR, HMBC, gNOE e EM.

The essential oil of Piper regnellii (Miq.) C. DC. var. pallescens (C. DC.) Yunck Piparaceae leaves, which were collected at a tree farm named Horto de Plantas Medicinais of the Universidade Estadual de Maringá, was obtained by hydrodistillation. A preliminary analysis by GC/MS was carried out. b-mirceno (70 percent) was identified as the main constituent by comparing MS and 13C NMR with the literature data. Four neolignans were isolated from the leaves and identified: eupomatenoid-6, eupomatenoid-5, eupomatenoid-3 and conocarpan. Their structures were established by extensive ¹H and 13C NMR, ¹H x ¹H - COSY, HETCOR, HMBC, gNOE and MS spectral studies.

Avaliação dos efeitos das raízes de Pfaffia glomerata (Spreng) Pedersen sobre o tempo de sono e crescimento bacteriano

Foram realizados testes biológicos com extrato hidroalcoólico liofilizado das raízes de Pfaffia glomerata. O teste de tempo de sono em camundongos envolveu administração de 50 mg/kg de pentobarbital sódico (via i.p.). O extrato foi administrado em várias doses agudamente e após 10 e 30 dias de tratamento oral. No teste antibacteriano, pequena quantidade do extrato de P. glomerata foi misturado com o inóculo de Staphylococcus aureus, Escherichia coli, Bacillius subtilis e Pseudomonas aeruginosa diluídos 1:10 para a microdiluição em caldo. No teste de tempo de sono, no modelo agudo obteve-se efeitos depressores para as doses de 500 e 1000 mg/kg; nos animais tratados cronicamente, aos 10 dias de tratamento obteve-se efeito depressor com 50 mg/kg e efeito estimulante com 500mg/kg; aos 30 dias de tratamento outro efeito estimulante ocorreu com a dose de 1 mg/kg. No teste antibacteriano não se observou inibição de crescimento de nenhuma das amostras das bactérias na maior concentração de 1000μg/ml do extrato. Conclui-se que o extrato das raízes de P. glomerata não tem efeito antimicrobiano e parece promover alguma interferência sobre o sono de animais de modo bifásico de acordo com as doses e tempos de tratamento.

Was made biological tests with hidroalcoolic extract of Pfaffia glomerata roots. In sleeping time test in mice, the extract was administered in several doses acutely and after 10 and 30 days of oral treatment. In the antibacterial analysis, the effect of the extract was evaluated on Staphylococcus aureus, Escherichia coli, Bacillius subtilis and Pseudomonas aeruginosa. In the sleeping time, in the acute model it was obtained depressive effects for the doses of 500 and 1000 mg/kg; in the chronically treated animals, to the 10 days of treatment it was obtained depressive effect with 50 mg/kg and stimulating effect with 500 mg/kg; to the 30 days of treatment other stimulating effect happened with the dose of 1 mg/kg. In the antibacterial test growth inhibition was not observed of none of the samples of the bacteria in the largest concentration of 1000μg/ml of the extract. It is ended that the extract of the roots of P. glomerata doesn't have antimicrobial effect and it seems to promote interference on the sleep of animals in a two-phase way in agreement with the doses and times of treatment.

Estudo físico-químico, químico e biológico de extrato das cascas de Stryphnodendron polyphyllum Mart. (Leguminosae)

Two flavan-3-ols and one proanthocyanidins have been isolated from the stem bark of Stryphnodendron polyphyllum Mart., which is traditionally used in Brazil against various diseases. The structure was determined on the basis of spectroscopic data including 1-D (¹H, 13C) and 2-D NMR (¹H/¹H COSY) and MS. The antibacterial activities of an acetone:water and semipurified extracts from the stem bark of Stryphnodendron polyphyllum Mart. were evaluated. Both the crude and semipurified extracts showed activity against Bacillus subtilis and Staphylococcus aureus. Quality control was determined using several pharmacopoeial assay.

Avaliação da atividade antibacteriana e antifúngica de extratos de plantas utilizados na medicina popular

Foram selecionados extratos de 13 plantas utilizadas na medicina popular brasileira para avaliar a atividade antimicrobiana. Destes, 10 extratos apresentaram níveis variados de atividade antibacteriana. Cinco dos extratos testados, apresentaram compostos com valores de Rf similares a de compostos antibacterianos visíveis na bioautografia. Três destas plantas pertencem à família Compositae indicando que o mesmo composto pode ser responsável pela atividade antibacteriana destas plantas. Atividade anticandida foi observada em 9 extratos de plantas. Os resultados podem explicar o uso etnobotânico das espécies estudadas para o tratamento de várias doenças infecciosas.

Extracts of 13 Brazilian medicinal plants were screened for their antimicrobial activity against bacteria and yeast. Of these, 10 plant extracts showed varied levels of antibacterial activity. Five of the plant extracts presented compounds with Rf values similar to the antibacterial compounds visible on bioautogram. Of these, three plants belong to the Compositae family. This may mean that the same compounds are responsible for the antibacterial activity in these plants. Anticandidal activity was detected in 9 plant extracts. The results might explain the ethnobotanical use of the studied species for the treatment of various infectious diseases.