Portrayal of Punica granatum L peel extract through High Performance Liquid Chromatography and antimicrobial activity evaluation / Retrato do extrato de casca de Punica granatum L por meio de cromatografia líquida de alto desempenho e avaliação da atividade antimicrobiana

ncreasing trend in antimicrobial resistance and failure of chemically synthesized antibiotics lead to discover alternative methods for the treatment of bacterial infections. Various medicinal plants are in use traditionally and their active compounds can be further applied for treatment of bacterial diseases. This study was designed to determine the antibacterial activity of Punica granatum (P. granatum L.) (pomegranate) peel extract against Enterobacteriaceae [Escherichia coli (E. coli), Salmonella Typhimurium (S. Typhimurium) and Shigella Dysenteriae (S. Dysenteriae)] and gram-positive bacterium [Staphylococcus aureus (Staph aureus)]. Methanolic extract of P. granatum L. peel was prepared by Soxhlet apparatus method. Total flavonoid and phenolic contents from the extract were determined by High Performance Liquid Chromatography (HPLC). The antibacterial activity of P. granatum L. peel extract was evaluated through agar well diffusion method. HPLC showed the range of phenolics (gallic acid, caffeic acid, benzoic acid, cinnamic acid) and flavonoid compounds. The chemical structures of flavonoid and phenolics found in the methanolic extract of P. granatum L. peel have been reported for the first time. The methanolic peel extract (50 ul) of yellow P. granatum L. showed 26, 10, 10 and 9mm zones of inhibition (ZOI) against S. aureus, S. Typhimurium, S. Dysenteriae and E. coli, respectively. The methanolic extract of red P. granatum L. (100 ul) showed 27, 8, 12 and 15 mm ZOI against Staph. aureus, S. Typhimurium, S. Dysenteriae and E. coli, respectively. Highest ZOI was observed against Staph. aureus. Many of the bacteria studied in the present work may cause serious gastrointestinal infections, which can lead to hemorrhagic diarrhea in children. These [...].

A tendência crescente na resistência antimicrobiana e na falha dos antibióticos sintetizados quimicamente leva à descoberta de métodos alternativos para o tratamento de infecções bacterianas. Várias plantas medicinais estão em uso tradicionalmente e seus compostos ativos podem ser posteriormente aplicados para o tratamento de doenças bacterianas. Este estudo foi desenhado para determinar a atividade antibacteriana do extrato de casca de Punica granatum (P. granatum L.) (romã) contra Enterobacteriaceae [Escherichia coli (E. coli), Salmonella Typhimurium (S. Typhimurium) e Shigella Dysenteriae (S. Dysenteriae) ] e bactéria gram-positiva [Staphylococcus aureus (Staph aureus)]. O extrato metanólico da casca de P. granatum L. foi preparado pelo método do aparelho de Soxhlet. O conteúdo total de flavonoides e fenólicos do extrato foi determinado por cromatografia líquida de alta eficiência (HPLC). A atividade antibacteriana do extrato da casca de P. granatum L. foi avaliada através do método de difusão em ágar. HPLC mostrou a gama de compostos fenólicos (ácido gálico, ácido cafeico, ácido benzoico, ácido cinâmico) e flavonoides. As estruturas químicas de flavonoides e fenólicos encontradas no extrato metanólico da casca de P. granatum L. foram relatadas pela primeira vez. O extrato metanólico da casca (50 ul) de P. granatum L. amarelo apresentou zonas de inibição (ZOI) de 26, 10, 10 e 9mm contra S. aureus, S. Typhimurium, S. Dysenteriae e E. coli, respectivamente. O extrato metanólico de P. granatum L. vermelho (100 ul) apresentou 27, 8, 12 e 15 mm IOI contra Staph. aureus, S. Typhimurium, S. Dysenteriae e E. coli, respectivamente. O ZOI mais alto foi observado contra Staph. aureus. Muitas das bactérias estudadas no presente trabalho podem causar infecções gastrointestinais graves, que podem levar à diarreia [...].

Effect of heavy metals on stem anatomical characteristics of Trapa natans L from Skadar Lake (Montenegro)

This study aimed to analyze stem anatomical characteristics of Trapa natans from five Skadar Lake locations (L1 - Milovica bay, L2 ­ inflow of the Moraca river, L3 - Kamenik, L4 - Grmozur, L5 - Lipovik) with different concentrations of Cu, Mn, Zn, Co and Pb, during the summer period of the year 2012. Cross sections of stem were made using cryotechnic procedure. For all analyzed quantitative anatomical parameters, the minimum values at the location L2 were recorded, with the presence of maximum content for all investigated metals in stem of Trapa natans. On the other hand, except for cobalt, at the location L4 the minimum concentration for all investigated metals were recorded. Plants collected from this location have the largest average values of the most measured anatomical parameters. The results of Discriminant Analysis showed that plants from different location could be clearly classified into three groups according to their stem quantitative anatomical characteristics which corresponds with heavy metals content. Our research also showed that there is no statistically significant correlation between the content of most investigated metals (Co, Cu, Pb and Zn) and the values of anatomical parameters. Statistically significant negative correlation was found between Mn content in stem and values of two anatomical characters (stem cross-sectional area and Mn content, r = - 0.88; p < 0.05; number of hypodermal cell layers and Mn content, r = - 0.90; p < 0.05).

In-vitro evaluation of Nigella sativa and Punica granatum effect on protoscolices of hydatid cysts / Avaliação in vitro do efeito de Nigella sativa e Punica granatum em protoescólices de cistos hidáticos

Abstract Cystic echinococcosis (CE) are commonly found in the liver and lungs of affected hosts. The treatment approach is usually surgical, or giving drugs in conjunction before surgery to kill protoscolices, to avoid anaphylactic shock from leakage of hydatid fluid into the peritoneum and to decrease opportunities for recurrences. The present study was to evaluate the in vitro scolicidal efficacy of hydroalcoholic extract of Punica granatum peel and Nigella sativa, on the protoscolices of CE that collected from the lungs of infected camels. Different concentrations of extracts with different exposure times were used and a viability assay was applied to measure the scolicidal effect. N. sativa showed its highest scolicidal efficacy at 100 mg/mL and 10 mg/mL concentrations after 30 and 60 min. P. granatum peel extract showed its maximum scolicidal efficacy at 100 mg/mL concentration after 120 min. All experiments of the current study revealed that the extracts of both N. sativa and P. granatum had a scolicidal effects on the protoscolices of camel hydatid cysts. It could be concluded that N. sativa extract is more potent than P. granatum peel extract regarding scolicidal effect, but the efficacies of both extracts were of moderate significant correlation to exposure time and concentrations.

Resumo Os cistos hidáticos (equinococose cística, CE) são comumente encontrados no fígado e nos pulmões dos hospedeiros afetados. A abordagem do tratamento geralmente é cirúrgica, e algumas drogas são administradas em conjunto antes da cirurgia para matar protoscólices e evitar choque anafilático devido ao vazamento de fluido hidático no peritônio e diminuir as oportunidades de recorrência. O presente estudo foi avaliar a eficácia in-vitro do extrato hidroalcoólico de casca de Punica granatum e Nigella sativa, sobre os protoescólices de cistos hidáticos, que foram coletados dos pulmões de camelos infectados. Concentrações dos extratos com diferentes tempos de exposição foram utilizados e um ensaio de viabilidade foi aplicado para medir o efeito escolicida. A N. sativa apresentou sua maior eficácia escolicida nas concentrações de 100 mg/mL e 10 mg/mL após 30 e 60 min. O extrato de casca de P. granatum mostrou sua máxima eficácia escolicida na concentração de 100 mg/mL após 120 min. Todos os experimentos do presente estudo revelaram que os extratos de N. sativa e P. granatum tiveram efeito escolicida dependente da dose e do tempo nos protoescólices dos cistos hidáticos de camelo. Pode-se concluir que o extrato de N. sativa é mais potente que o extrato de casca de P. granatum em relação ao efeito escolicida, mas a eficácia de ambos os extratos foi de correlação significativa moderada com o tempo de exposição e as concentrações.

Pollen/Fruit Syndrome: Clinical Relevance of the Cypress Pollen Allergenic Gibberellin-Regulated Protein

A pollen/food-associated syndrome (PFAS) has been described between peach and cypress pollen. Cross-reactive allergens were characterized which belong to the Gibberellin-regulated protein (GRP) family, BP14 in cypress pollen and Pru p 7 in peach. GRP are small cationic protein with anti-microbial properties. A patient suffering from a peach/cypress syndrome was explored clinically and biologically using 2 types of immunoglobulin E (IgE) multiarray microchip, immunoblots and a basophil activation test to assess the clinical relevance of various extracts and purified allergens from fruits or cypress pollen. In addition to PR10 sensitization, the patient showed specific IgE to Pru p 7, BP14 and allergen from pomegranate. These last 3 allergens and allergenic sources are able to induce ex vivo basophil activation characterized by the monitoring of the expression of CD63 and CD203c, both cell surface markers correlated with a basophil mediator release. Up to 100% of cells expressed CD203c at 50 ng/mL of BP14 protein. In contrast, snakin-1, a GRP from potato sharing 82% sequence identity with Pru p 7 did not activate patient's basophils. These results strongly suggest that, like Pru p 7, BP14 is a clinically relevant allergenic GRP from pollen. Allergen members of this newly described protein family are good candidates for PFAS where no cross-reactive allergens have been characterized.

Efecto diurético de Cuphea glutinosa Cham. et Schltdl (Lythraceae) en ratas Wistar / Diuretic effect of Cuphea glutinosa Cham. et Schltdl (Lythraceae) in Wistar rats

Cuphea glutinosa (Lythraceae), popularmente conocida como "siete sangrías" o "sanguinaria", ha sido utilizada en las prácticas de medicina popular, especialmente como diurética e hipotensora. Actualmente, es comercializada en herboristerías de la República Argentina y consumida en forma de infusión en concentraciones no específicas. En este estudio se evaluó la diuresis producida en ratas Wistar, a partir de la administración de infusiones de diferentes concentraciones de planta entera de C. glutinosa. Se utilizaron tres concentraciones al 2,5 %; 5,0 % y 10,0 %, Furosemida (10 mg/kg peso vivo) como control positivo y solución salina (blanco) (NaCl 0,9 %) como control negativo, que fueron administradas oralmente. Se midieron los niveles de excreción urinaria de las ratas, mantenidas en jaulas metabólicas, en intervalos de 2 horas, durante 8 horas, y una última recolección a las 24 horas. La administración oral de infusión al 5 % incrementó significativamente la excreción urinaria a las 8 horas, comparada con el control positivo. Además se observó una tendencia a la diuresis con la administración de infusiones al 5 % y 10 % a las 4 horas. Durante la aplicación de los tratamientos no se observaron cambios en las condiciones físicas de los animales. Se requieren nuevos estudios para la determinación de la ruta farmacológica y la toxicidad de esta especie. (AU)

Supplementing punicalagin reduces oxidative stress markers and restores angiogenic balance in a rat model of pregnancy-induced hypertension

Pre-eclampsia (PE) is a pregnancy disorder that is characterised by severe hypertension and increased risks of foetal and maternal mortality. The aetiology of PE not completely understood; however, maternal nutrition and oxidative stress play important roles in the development of hypertension. The treatment options for PE are currently limited to anti-hypertensive drugs. Punicalagin, a polyphenol present in pomegranate juice, has a range of bioactive properties. The effects of supplementation with punicalagin on angiogenesis and oxidative stress in pregnant rats with induced hypertension were investigated. The pregnant rats were randomly divided into five experimental groups (n=12 per group). Hypertension was induced using an oral dose of NG-nitro-L-arginine methyl ester (L-NAME, 50 mg/kg/day) on days 14–19 of pregnancy. Punicalagin (25, 50 or 100 mg/kg) was given orally on days 14–21 of pregnancy. Punicalagin treatment at the tested doses significantly reduced diastolic, systolic, and mean arterial blood pressure in L-NAME treated rats from day 14. Punicalagin also restored angiogenic balance by increasing the expression of vascular endothelial growth factor and downregulating vascular endothelial growth factor receptor-1/fms-like tyrosine kinase-1. Punicalagin, significantly increased the placental nitric oxide levels as compared to PE group. The increased levels of oxidative stress in rats with PE were markedly decreased by treatment with punicalagin. Punicalagin at the tested doses markedly (p < 0.05) enhanced the placental antioxidant capacity in L-NAME-treated rats. The raised catalase activity observed following L-NAME induction was significantly (p < 0.05) and restored to normal activity levels in punicalagin treatment. Further, 100 mg dose of punicalagin exhibited higher protective effects as compared to lower doses of 25 and 50 mg. This study shows that supplementation with punicalagin decreased blood pressure and oxidative stress and restored angiogenic balance in pregnant rats with induced PE.

Fitossíntese de nanopartículas de prata a partir de extrato de cascas de romã e desenvolvimento de formulação para tratamento de feridas: avaliação antimicrobiana, citotóxica e potencial cicatrizante em um modelo in vivo / Phytosynthesis of silver nanoparticles using pomegranate peel extract and development of formulation for wound healing: antimicrobial and cytotoxicity evaluation and repair potential in a in vivo model

O objetivo deste estudo foi avaliar a capacidade de produção de nanopartículas de prata através do extrato da casca de romã, e produzir formulações contendo estas nanopartículas para uso em feridas. Elas foram testadas quanto à ação antimicrobiana, citotóxica e potencial cicatrizante. Para a produção das nanopartículas de prata propôs-se uma síntese utilizando-se como base duas metodologias já estabelecidas na literatura, utilizando-se para reação carboximetilcelulose, propilenoglicol, nitrato de prata, água e extrato da casca de romã como agente redutor. O extrato da casca de romã foi caracterizado em parâmetros como pH, massa seca e quantidade de taninos bioativos (ácido elágico e totais fenólicos expressos em ácido gálico). Os totais fenólicos do extrato foram também dosados após seu aquecimento nas diferentes condições de tempo e temperatura propostos para as sínteses das nanopartículas de prata (12 minutos, 1 hora e 2 horas, à 50ºC e 100ºC). As nanopartículas de prata produzidas foram, então, adicionadas a uma solução contendo compostos para produção de formulações para serem utilizadas no tratamento de feridas. Elas foram caracterizadas através de espectroscopia UV-Visível, microscopia eletrônica de varredura (MEV), potencial zeta e dosagem de íons remanescentes após as reações. A atividade antimicrobiana tanto das nanopartículas como de suas formulações contra Candida albicans SC 5314 e Staphylococcus aureus ATCC 25923 foi avaliada por meio do método da microdiluição. Na avaliação dos extratos, apesar de ocorrer um aumento na concentração de totais fenólicos com o aumento da temperatura, a concentração inibitória mínima (CIM) manteve-se estável com valores de 391 µg/ml e 781 µg/ml para S. aureus e C. albicans. A formação das nanopartículas de prata foi confirmada com a formação de picos característicos na espectroscopia UV-Visível e pelas imagens de MEV, onde verificou-se que a síntese com tempo de reação de 12 minutos e aquecimento a 50ºC gerou nanopartículas mais uniformes e melhor distribuídas na formulação. As sínteses propostas promoveram a redução iônica da prata de aproximadamente 100%, independente do tempo temperatura utilizados na reação. Valores de CIM para as nanopartículas de prata foram de 67,50 µg/ml e 68,75 µg/ml respectivamente para S. aureus e C. albicans independente das variações das condições de síntese. Após a seleção da síntese das nanopartículas de prata por 12 min à 50ºC, estas partículas foram também caracterizadas por difração de raios-X e microscopia eletrônica de transmissão (TEM), e as respectivas formulações por meio de espalhamento de luz dinâmica, dosagem de íons livres, potencial zeta, MEV e TEM. Para essas formulações foi também realizado um teste de estabilidade variando-se umidade e temperatura. Além da atividade antimicrobiana contra Candida albicans SC 5314 e Staphylococcus aureus ATCC 25923, a citotoxicidade em fibroblastos (L929) das nanopartículas de prata e das formulações destas partículas e do extrato da casca de romã foram também avaliadas. Para os extratos foram observados valores de 3,13, 86,39±0,96% m/m, 3,64±0,03 mg/g, 392,0±9 mg/g respectivamente para pH, massa seca, ácido elágico e totais fenólicos. Como controle, foram produzidas nanopartículas de prata sintetizadas convencionalmente (AgNP química), e observou-se potencial redutor de 99,89% e 99,51% para as sínteses utilizando-se extrato de romã (AgNP green) e um agente redutor químico convencional (AgNP química). Verificou-se a formação de partículas com tamanhos médios de 89 e 19 nm para nanopartículas green e química. A formulação contendo as nanopartículas de prata apresentaram um potencial antimicrobiano expressivamente maior do que o princípio ativo isolado, sendo 255 e 4 vezes mais efetiva contra S. aureus e C. albicans, respectivamente. Os valores de citotoxicidade foram consideravelmente menores para as nanopartículas de prata sintetizadas pelo extrato de romã quando comparadas as produzidas convencionalmente. De acordo com os valores de CIM e da citotoxicidade, a concentração das formulações foi ajustada, gerando assim três formulações: i) AgNP green, ii) AgNP química e iii) extrato de romã, nas concentrações de 337,5 µg/ml, 5,55 µg/ml e 94 µg/ml respectivamente. Para o estudo in vivo, utilizou-se como controle um medicamento comercial contendo prata indicado para tratamento de feridas (Sulfadiazina de prata). Foram selecionados noventa ratos Wistar machos com peso médio de 180 gramas. Foi induzida diabetes nos ratos, e, em seguida, realizou-se duas incisões no dorso dos animais e as lesões foram imediatamente infectadas com S. aureus (ATCC 25923) e C. albicans (SC 5314). Após 24 h, os animais foram divididos em grupos de acordo com as formulações propostas em cada tratamento, seguindo-se um protocolo de duas vezes ao dia por 2, 7 e 14 dias. Após o período de tratamento os animais foram eutanasiados e verificado o potencial reparador através do índice de fechamento de ferida, avaliação do infiltrado inflamatório, angiogenese, mieloperoxidase e hidroxiprolina. Ainda foi verificado o potencial antimicrobiano das formulações através da contagem de células viáveis de cada microrganismo infectado nas feridas. De forma geral, as formulações contendo nanopartículas de prata mostraram os melhores resultados para o fechamento das feridas, apresentando ainda uma atividade anti-inflamatória maior que a do extrato de casca de romã, o qual apresentou atividade pró inflamatória. Todos os tratamentos não foram capazes de reduzir de forma significativa o número de células de C. albicans, enquanto para S. aureus todos os tratamentos apresentaram redução significativa após quatorze dias de tratamento. Independente das nanopartículas de prata serem produzidas quimicamente ou por meio do extrato da casca de romã, ambas apresentaram considerável potencial de reparo de feridas infectadas em modelos in vivo com ratos. Os achados das presentes pesquisas reforçam e estimulam o uso potencial das nanopartículas de prata no tratamento de feridas, com destaque para a síntese green por gerar menos danos ao meio ambiente e às pessoas envolvidas tanto em sua produção como aos pacientes, e por apresentar, ainda, custo inferior quando comparada às sínteses utilizando reagentes químicos e processos convencionais(AU)

The aim of this study was to investigate the production of silver nanoparticles through peel extract of pomegranate, and produce formulations containing these particles to be used in wound healings. Its antimicrobial action, cytotoxicity and healing potential were tested. The synthesis of silver nanoparticles were based on two methods proposed in the literature with some modifications, which were used carboxymethylcellulose, propylene glycol, silver nitrate, water and peel extract of pomegranate as reducing agent. The peel extract was characterized by pH, dry mass and bioactive tannins (elagic acid and total phenols expressed as galic acid). The total phenols were also quantified after being heated at 50ºC and 100ºC for 12 minutes, 1 hour and 2 hours. Then, silver nanoparticles were added in a solution containing products to develop a formulation to be tested in wound healing. They were characterized by UV-Vis spectroscopy, scanning electron microscopy (SEM), zeta potential and the quantification of remaining silver ions after the synthesis reaction. The antimicrobial activity of the nanoparticles and formulations were tested against Candida albicans (SC 5314) e Staphylococcus aureus (ATCC 25923) by microdilution method. After submitting the peel extract to different conditions of temperature and times (50ºC and 100ºC for 12 minutes, 1 hour and 2 hours), it was noted that the values of the minimum inhibitory concentration was not affected and were 391 µg/ml and 781 µg/ml for S. aureus and C. albicans. The formation of silver nanoparticles was confirmed through the formation of characteristic peaks in the UV-Vis spectroscopy and SEM images, and it was observed that the reaction at 50ºC for 12 min produced silver nanoparticles with regular forms and better dispersed in the formulation. The synthesis proposed promoted the reduction of silver ions at about 100%, regardless of the time and temperature used in the reaction, which also did not interfere in antimicrobial activity against C. albicans (68,75 µg/ml) and S. aureus (67,50 µg/ml). After selecting the reaction at 50ºC for 12 min, the silver nanoparticles produced ere also characterized by X-ray diffraction and transmission electron microscopy (TEM), and the respective formulations through dynamic light scattering, free ion dosage, zeta potential, SEM and TEM. The stability test varying humidity and temperature was also performed for those formulations. Besides antimicrobial assays, the cytotoxicity (L929 fibroblasts) of the silver nanoparticles and the formulations of these particles and of the pomegranate peel extract were evaluated. It was observed in the peel extract the values of 3,13, 86,39±0,96% m/m, 3,64±0,03 mg/g and 392,0±9 mg/g respectively for pH, dry mass, elagic acid and total phenols concentrations. Silver nanoparticles produced by conventional chemical method was prepared and used as controls, and it was noted the reduction potential of 99,89% and 99,51% for the synthesis using pomegranate peel extract (AgNP green) and chemical reducing agent (AgNP chemical). The averages sizes of green and chemical AgNP were 89 and 19 nm. The formulation containing silver nanoparticles presented an antimicrobial potential expressively higher than the active input, being 254 and 5- fold more effective against S. aureus and C. albicans. Also, the cytotoxicity was notable reduced when silver nanoparticles were produced using pomegranate peel extract. Based on the MIC values and the cytotoxicity findings, the concentration of the formulations were determined: i) AgNP green at 337.5 µg/ml, ii) AgNP chemical at 5.55 µg/ml, and iii) pomegranate peel extract at 94 µg/ml. In the in vivo study, a commercial form of silver (Sulfadizsine) to the wound healing was used as control. Ninety Wistar male rats were selected, and, after inducing diabetes, two incisions on the dorsum of the animals were made and followed infected with S. aureus (ATCC 25923) and C. albicans (SC 5314). After the infection, the animals were treated with the formulations twice a day for 2, 7 and 14 days. Then, the animals were euthanized and the repair potential was verified through wound closure index, inflammatory infiltrate evaluation, angiogenesis, myeloperoxidase and hydroxyproline. It was also determined the antimicrobial potential by counting the viable cells of each microorganism used to infect the wounds. In general, the formulations containing silver nanoparticles promoted a better closure of the wounds, and a higher anti-inflamatory activity than the peel extract formulation which otherwise presented a pro-inflamatory effect. All formulations could not significantly reduce the viable cells of C. albicans, while for S. aureus they reduced significantly the cells after 14 days of treatment. Silver nanoparticles produced by both green and conventional chemical process present notable potential in repairing infected wounds in in vivo rat model. The findings of the present research strengthen and stimulate the potential application of silver nanoparticles in wound healings, highlighting the green production of these particles which apart from being lower costly, it is ecofriendly and less detrimental to people involved in its production and use(AU)

Pomegranate leaf attenuates lipid absorption in the small intestine in hyperlipidemic mice by inhibiting lipase activity / 中国天然药物

Pomegranate leaf (PGL) has a definite role in regulating lipid metabolism. However, pharmacokinetic results show the main active ingredient, ellagic acid, in PGL has lower oral bioavailability, suggesting that the lipid-lowering effect of PGL may act through inhibiting lipid absorption in the small intestine. Our results demonstrated that pomegranate leaf and its main active ingredients (i.e., ellagic acid, gallic acid, pyrogallic acid and tannic acid) were capable of inhibiting pancreatic lipase activity in vitro. In computational molecular docking, the four ingredients had good affinity for pancreatic lipase. Acute lipid overload experiments showed that a large dosage of PGL significantly reduced serum total cholesterol (TG) and triglycerides (TC) levels in addition to inhibiting intestinal lipase activity, which demonstrated that PGL could inhibit lipase activity and reduce the absorption of lipids. We also found that PGL could reverse the reduced tight-junction protein expression due to intestinal lipid overload, promote Occludin and Claudin4 expression in the small intestine, and enhance the intestinal mucosal barrier. In conclusion, we demonstrated that PGL can inhibit lipid absorption and reduce blood TG and TC by targeting pancreatic lipase, promoting tight-junction protein expression and thereby preventing intestinal mucosa damage from an overload of lipids in the intestine.

Pomegranate extracts on larval inhibition of Haemonchus spp and Cooperia spp obtained from cattle / Extratos de romã sobre inibição larval de Haemonchus spp e Cooperia spp provenientes de bovinos

Consumers increasingly demand natural food and absent of toxic waste and medicated. The aim of this study was to evaluate the aqueous extract of dried and in natura stem from pomegranate on the larval inhibition of Haemonchus spp. and Cooperia spp. obtained from cattle. Dried and in natura aqueous extract from pomegranate stem (AEPS), negative control (autoclaved distilled water) and positive control (levamisole hydrochloride 0.05 mg / mL) were evaluated. Four mL of each treatment were used, and the results were evaluated in triplicate by egg counts per gram of feces (EPG), larval culture and counting of the larvae. The mean of the larvae that developed per gram of feces was submitted to logarithimic transformation log. (X + 10). The EPG had an average value of 3.620 eggs, identified in larval culture from Haemonchus spp. and Cooperia spp. Dried AEPS presented, by the reduction of larvae, effectiveness of 92.29% for Haemonchus spp. and 96.97% for Cooperia spp. In natura, this extract showed 80% of efficacy against Cooperia larvae, being considered moderately effective; however, for Haemonchus spp. this extract was not effective. These results are specially important for organic and agroecological systems of production, that not allow the use of chemical parasiticides. In conclusion, the use of dried AEPS showed potential anthelmintic and may represent important natural therapeutic option.

Os consumidores têm exigido cada vez mais alimentos naturais e ausentes de resíduos medicamentosos. O objetivo deste estudo foi avaliar o extrato aquoso de caule de romã seco e in natura, sobre a inibição larval de Haemonchus spp. e Cooperia spp. provenientes de bovinos. Foram avaliados extrato aquoso de caule de romã (EACR) seca e in natura, controle negativo (água destilada autoclavada) e controle positivo (cloridrato de levamisol 0,05 mg/mL). Foram utilizados quatro mL de cada tratamento, e os resultados foram avaliados em triplicata, por meio de contagem de ovos por grama de fezes (OPG), cultura de larvas e contagem das larvas. A média de larvas desenvolvidas por grama de fezes foi submetida à transformação logarítmica Log. (x+10). A contagem de OPG apresentou valor médio de 3.620 ovos, sendo identificadas na cultura de larvas de Haemonchus e Cooperia. EACR seca apresentou por meio da redução de larvas, eficácia efetiva de 92,29% para Haemonchus e 96,97% para Cooperia. In natura, este extrato apresentou 80% de eficácia sobre larvas de Cooperia, sendo considerado moderadamente efetivo, porém, para Haemonchus o mesmo não foi efetivo. Esses resultados são especialmente importantes para os sistemas orgânicos e agroecológicos de produção, que não permitem a utilização de parasiticidas químicos. Concluiu-se que a utilização de EACR seca apresentou potencial anti-helmíntico, podendo representar importante opção terapêutica natural.

Phytochemical profiling and bioactivity of a mangrove plant, Sonneratia apetala, from Odisha Coast of India / 中国结合医学杂志

<p><b>OBJECTIVE</b>To test the antioxidant, antidiabetic, anticancer and antibacterial activities along with phytochemicals of Sonneratia apetala Buch.-Ham.</p><p><b>METHODS</b>The antibacterial activity was determined by agar well diffusion method. The antioxidant activity was determined by standard assay. The antidiabetic activity was evaluated by α-glucosidase inhibition assay and in vivo anticancer property was determined against Ehrlich ascites carcinoma (EAC) cells in Swiss Albino mice. Further partial characterization of the methanol extracts was carried out by thin layer chromatography, high performance liquid chromatography, (1)H nuclear magnetic resonance spectroscopy and Fourier transform-infra red spectrum spectral analysis.</p><p><b>RESULTS</b>Four solvent extracts (acetone, ethanol, methanol and aqueous) of leaf and bark possess strong antioxidant properties. In vivo anticancer activity of methanol extract leaf indicated positive activity showing 34% inhibition against EAC cells in Swiss Albino mice. All extracts exhibited α-glucosidase inhibitory activity in a dose-dependent manner indicating presence of promising antidiabetic properties. The extracts possess strong antibacterial activity against the selected pathogenic bacteria (minimal inhibitory concentration ranging from 1.25-5.00 mg/mL). The partial characterization of the methanol extracts of leaf and bark revealed the presence of phenolics as the lead compound responsible for studied bioactivities of the plant extracts.</p><p><b>CONCLUSION</b>Sonneratia apetala extracts have potent antibacterial, antioxidant, antidiabetic and anticancer properties which can be further exploited for its pharmaceutical applications.</p>

Avaliação in vitro do potencial antimicrobibiano do decoto de cascas de romã (Punica granatum L. ) sobre amostras de saliva de escolares / In vitro assessment of the antimicrobial effects of pomegranate (Punica granatum L.) peel decoction on saliva samples

Existem vários esforços para o desenvolvimento de produtos capazes de reduzir ou eliminar os microrganismos patogênicos presentes na cavidade oral. A literatura relata uma série de efeitos adversos associados ao uso contínuo destes produtos, dentre eles vômitos, diarreia e o escurecimento da dentina. A indução da resistência microbiana é um dos fatores de destaque relacionado ao uso destes produtos. Neste trabalho, o decocto de romã (Punica granatum L.), obtido a partir das cascas do fruto, foi utilizado para avaliação de seu potencial antimicrobiano sobre cepas de Pseudomonas aeruginosa, Candida albicans e Enterococcus faecalis, sendo ativos contra os dois primeiros microrganismos. A aplicação do decocto sobre os microrganismos presentes em amostras de saliva de crianças mostrou halos de inibição semelhantes ao obtido com a solução de clorexidina a 0,12%. A atividade antimicrobiana do decocto de romã aponta esta preparação como uma fonte em potencial para o desenvolvimento de produtos de uso oral...

Several products have been developed to eliminate or reduce potential pathogenic microorganisms of the oral microbiome. The continuous use of these synthetic products can result in side effects such as vomiting, diarrhea, darkening of the teeth and the induction of microbial resistance. Pomegranate (Punica granatum) peel decoction was tested to assess its antimicrobial activity. In vitro analysis showed the decoction had antimicrobial activity against strains of Pseudomonas aeruginosa and Candida albicans, but none was detected against Enterococcus faecalis. When tested on saliva samples from children, the decoction showed great potential in reducing the load of microorganisms, the inhibition haloes produced with saliva samples being similar to those of the antimicrobial control (0.12% chlorhexidine). The pomegranate peel decoction in water could thus provide a promising source for developing solutions for use against oral diseases...

Phenol content, antioxidant and tyrosinase inhibitory activity of mangrove plants in Micronesia

OBJECTIVE@#To find out and compare the in vitro antioxidant and tyrosinase inhibitory activities of two species of mangrove plants.@*METHODS@#Mangrove samples were harvested at the shoreline on the island of Weno, Chuuk State in Micronesia. The phenol content, antioxidant activity (based on DPPH-free radical scavenging) and tyrosinase inhibitory activity in different tissues (leaves, barks and roots) of Rhizophora stylosa (R. stylosa) and Sonneratia alba (S. alba), collected from the island of Weno.@*RESULTS@#Total phenol content ranged from 4.87 to 11.96 mg per g of freeze dried samples. The highest antioxidant activity was observed in R. stylosa bark (85.5%). The highest tyrosinase inhibitory activity was found in S. alba bark. Also, total phenol content and antioxidant activity were higher in methanol extracts than in aqueous extracts.@*CONCLUSIONS@#Taken together, the results of this study proved that mangroves can be excellent sources of antioxidant compounds.

Antifungal activity of the ethanolic extracts of Punica granatum L. and evaluation of the morphological and structural modifications of its compounds upon the cells of Candida spp.

Ethanolic crude extracts prepared from the arils and seeds, pericarp, peels and from the whole fruit of Punica granatum, known as pomegranate, had their antifungal activity tested against Candida spp. The ethanolic crude extracts were analyzed by Mass Spectrometry and yielded many compounds such as punicalagin and galladydilacton. The extracts from the pericarp and peel showed activity against Candida spp., with MICs of 125 µg/mL. The effect of pericarp and peel extracts upon the morphological and structure of C. albicans and C. krusei were examined by scanning and transmission electron microscopy, with the visualization of an irregular membrane and hyphae, formation of vacuoles and thickening of the cell wall. The data obtained revealed potential antimicrobial activity against yeasts cells of the Candida genus, and the bioactive compounds could be responsible for changes in cell morphology and structure. The data obtained open new perspectives for future research in continuation to this study, where information such as determination of the site of action of the compounds could contribute to an alternative therapy against these organisms.

Atividade antimicrobiana in vitro de extrato de Punica granatum L. sobre Staphylococcus aureus isolado em leite bovino / Antimicrobial activity in vitro of extracts of Punica granatum L. over Staphylococcus aureus isolated in bovine milk

Mastite bovina é considerada principal doença causadora de grandes perdas econômicas nos rebanhos leiteiros. Antimicrobianos químicos promovem resistência farmacológica, resíduos no alimento e contaminação ambiental. Este estudo objetivou verificar a atividade antibacteriana, in vitro, do extrato de romã em Staphylococcus aureus isolado de leite bovino, avaliar sua atividade antioxidante, e quantificar teores de fenóis e flavonoides totais nos diferentes extratos utilizados. Utilizou-se para tanto extratos aquosos da casca do fruto (EAC) e folhas (EAF), secos e in natura de romã. Adicionalmente, avaliou-se atividade antioxidante (AA%), teores de fenóis e flavonóides totais. Amostras de leite foram semeadas, incubadas, e as colônias de Staphylococcus aureus foram ajustadas a 1,0x106 UFC/mL ao padrão nº6 da escala de MacFarland. A sensibilidade dos isolados microbianos foi determinada, em quintuplicata, pela técnica de difusão em discos. A Concentração Inibitória Mínima foi determinada pelo halo de inibição superior a 15 mm. Os resultados foram avaliados pelo método ANOVA, teste de Tukey 5%, utilizando-se o programa SISVAR 5.3 - DEX/UFLA. EACseco inibiu crescimento bacteriano a partir de 3%. EAFseco, EACin natura e EAFin natura, só apresentaram esta atividade a partir de 15%, 20% e 30% respectivamente. A ação antioxidante do EACseco foi verificada a partir de 75µg/mL, com valores correspondentes a 14,2%, atingindo um platô de 65,9% na concentração de 250µg/mL. Entretanto, essa atividade não foi correlacionada aos teores de fenóis totais e flavonoides. Provavelmente outras substâncias alcalóides, podem ter sido responsáveis por esta atividade. Conclui-se que os extratos de Punica granatum L., principalmente aquele obtido pela casca do fruto seco, demonstraram atividade inibitória sobre S. aureus, evidenciando a potencialidade de seu uso para o controle da mastite bovina.

Bovine mastitis is considered important disease causing major economic losses in dairy herds. Antimicrobial drug can promote resistance, chemical residues in food and environmental contamination. The purpose of this study was to evaluate the antibacterial activity in vitro of pomegranate extract on Staphylococcus aureus isolated from bovine milk, evaluate its antioxidant activity, and quantify levels of total phenols and fravonoides of the different extracts used. Aqueous extracts were used in nature and dry, from the peel of the fruit (EAC) and leaves (EAF). Additionally, it was evaluated the antioxidant activity (AA%), total phenols and flavonoids. Milk samples were inoculated, incubated, and the colonies were characterized as Staphylococcus aureus were adjusted to 1.0x106 UFC/mL in the 6 standard of the MacFarland scale. The sensitivity of the microbial isolates were determined in quintuplicate, in disk diffusion test. The minimum inhibitory concentration was determined by visible inhibition zone greater than 15mm. The results were evaluated by ANOVA, Tukey 5%, using the program SISVAR 5.3 - DEX / UFLA. Results implied that the aqueous extract from the bark of dried fruit was capable of inhibiting bacterial growth at concentrations of 3%. The other treatments only showed this activity, from the concentrations of 15%, 20% and 30% for dry EAF, in nature EAC and in nature EAF, respectively. Regarding the action antioxidant of dry EAC, was not correlated with total phenols and flavonoids. Probably other alkaloids substances present in the extract studied, may have been responsible for this activity. It is concluded that extracts of Punica granatum L., especially those obtained by the shell of the fruit, showed inhibitory activity against S. aureus, indicating your use potential for the control of bovine mastitis.

Effect of pomegranate peel polyphenol gel on cutaneous wound healing in alloxan-induced diabetic rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Pomegranate (punica granatum) belongs to the family Punicaceae, and its peel has been used as a traditional Chinese medicine because of its efficacy in restraining intestine, promoting hemostasis, and killing parasites. Pomegranate peel has been reported to possess wound-healing properties which are mainly attributed to its polyphenol extracts. The purpose of this study was to investigate the effect of pomegranate peel polyphenols (PPP) gel on cutaneous wound healing in diabetic rats.</p><p><b>METHODS</b>Alloxan-induced diabetic rats were given incisional wounds on each side of the mid-back and then treated daily with PPP gel (polyphenol mass fraction = 30%) post-wounding. Rats were sacrificed on days 4, 7, 14, and 21 post-wounding to assess the rates of wound closure, histological characteristics; and to detect the contents of hydroxyproline, production of nitric oxide (NO), and activities of NO synthase (NOS), as well as the expressions of transforming growth factor-β1 (TGF-β1), vascular endothelial growth factor (VEGF), and epidermal growth factor (EGF) in wound tissue.</p><p><b>RESULTS</b>Wound closure was significantly shortened when PPP gel was applied to the wounds of diabetic rats. Histological examination showed the ability of PPP gel to increase fibroblast infiltration, collagen regeneration, vascularization, and epithelialization in the wound area of diabetic rats. In addition, PPP gel-treated diabetic rats showed increased contents of hydroxyproline, production of NO, and activities of NOS and increased expressions of TGF-β1, VEGF, and EGF in wound tissues.</p><p><b>CONCLUSION</b>PPP gel may be a beneficial method for treating wound disorders associated with diabetes.</p>

Effects of pomegranate flowers polyphenols on liver PON expression of diabetes combining non-alcoholic fat liver diseases rats / 药学学报

To investigate the protective effects and possible mechanism of pomegranate flowers polyphenols (PFP) on liver function of rats with diabetes combining non-alcoholic fat liver diseases, diabetes combining nonalcoholic fat liver disease model rats were established with high calorie feeding and small dose intraperitoneal injection of streptozotocin (STZ). Model rats were randomly divided into: model group, metformin group, pomegranate flowers polyphenols low, medium and high dose group (75, 150 and 300 mg x kg(-1)). After four weeks treatment, the levels of FPG, blood fat profiles and serum insulin, ALT, AST levels, SOD and MDA in the liver and serum separately were analyzed with biochemical methods. Paraoxonase (PON1 and PON3) mRNA and protein expression in liver were checked by RT-PCR and immunohistochemical method. Pathological changes of the liver were observed. FPG, IRI, non-HDL-C and transaminase significantly reduced and HDL-C raised in the each PFP dose group; Furthermore, compared with model group, fat drops in liver cells significantly reduced, antioxidant ability enhanced, PON1 mRNA and protein expression level in liver increased significantly. The protective effects of PFP against diabetes combining non-alcoholic fat liver diseases rats might through the increase liver PON1 mRNA and protein expression further enhanced the body antioxidant capacity and reduced IRI so as to ameliorate the rat hepatic steatosis.

Comparative study of chemical composition of pomegranate peel pomegranates inside and pomegranate seeds / 中国中药杂志

An HPLC fingerprint of pomegranate peel was established. Using chromatographic conditions, we compared the chemical composition of pomegranate peel, inside and seeds, and simultaneously determined the contents of gallic acid and ellagic acid. By comparison, we found that there were no significant differences between pomegranate peel and inside, but there was a big difference between pomegranate seeds and another two. The contents of gallic acid and ellagic acid of pomegranate peel respectively were 0.33%, 0.59%, while in pomegranate inside the result respectively were 0.52%, 0.38%. Content of ellagic acid from pomegranate seeds was only 0.01%. By study, we thought that when pomegranate peel was processed, pomegranate seeds should be removed, while pomegranate inside could be retained on the premise of full drying.

Oxidation of fatty acid may be enhanced by a combination of pomegranate fruit phytochemicals and acetic acid in HepG2 cells

We investigated whether the combination of phytochemicals and acetic acid in the form of fruit vinegar provides an additive effect on changes of mRNA levels related to fatty acid oxidation in human hepatocyte (HepG2). Among the seven fruit vinegars (Rubuscoreanus, Opuntia, blueberry, cherry, red ginseng, mulberry, and pomegranate) studied, treatment of HepG2 with pomegranate vinegar (PV) at concentrations containing 1 mM acetic acid showed the highest in vitro potentiating effect on the mRNA expression levels of peroxisome proliferator-activated receptor alpha, carnitinepalmitoyl transferase-1, and acyl-CoA oxidase compared to the control group (P < 0.05). Reversed-phase liquid chromatography in combination with quadrupole time-of-flight mass spectrometry analysis revealed four potential compounds (punicalagin B, ellagic acid, and two unidentified compounds) responsible for altered gene expression in HepG2 cells treated with PV as compared with the others. Further investigations are warranted to determine if drinking PV beverages may help to maintain a healthy body weight in overweight subjects.

Protective effects of pomegranate polyphenols on cardiac function in rats with myocardial ischemia/reperfusion injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To determine the protective effect of pomegranate polyphenols on cardiac function in rats with myocardial ischemia/reperfusion (I/R) injury and explore the possible mechanism.</p><p><b>METHODS</b>Fifty SD rats were randomized into 5 equal groups, including a sham-operated group, an I/R model group, and 3 pomegranate polyphenol dose groups (150, 300, and 600 mg/kg). The rats were subject to a 45-min left main coronary artery occlusion followed by a 180-min reperfusion to induce myocardial I/R injury except for the rats those in the sham-operated group. The cardiac functions were monitored continuously during the experiment. At the end of the reperfusion, arterial blood samples were obtained to measure plasma contents of lactate dehydrogenase (LDH), creatine kinase (CK), malondialdehyde (MDA) and the activities of superoxide dismutase (SOD). Triphenyltetrazolium chloride (TTC) staining was used to evaluate the infarct size.</p><p><b>RESULTS</b>Compared with the sham-operated group, all the rats with I/R showed significantly decreased left ventricular systolic pressure (LVSP), maximal rates of increase/decrease of the left ventricular pressure (±dp/dtmax) (P<0.01) and significantly increased left ventricular end-diastolic pressure (LVEDP) (P<0.01). Compared with the I/R model group, all the 3 pomegranate polyphenol groups had significantly improved cardiac function (P<0.05), decreased plasma contents of CK, LDH and MDA (P<0.01), increased SOD activities (P<0.01), and obviously reduced infarct size (P<0.01).</p><p><b>CONCLUSION</b>Pomegranate polyphenols can protect the cardiac function of rats with I/R injury probably in association with their actions in enhancing oxygen free radical scavenging activity and decreasing lipid peroxidative damage of the myocardial tissues.</p>