Interference of aqueous and ethanolic solutions of Adiantum latifolium Lam. (Pteridaceae) leaves on in vitro Ceratocystis cacaofunesta mycelial growth / Interferência de soluções aquosas e alcoólicas de folhas de Adiantum latifolium Lam. (Pteridaceae) no crescimento micelial de Ceratocystis cacaofunesta in vitro

Ceratocystis cacaofunesta is the etiologic agent of "Ceratocystis wilt of cacao", an irreversible disease that affects the vascular system of the plant. The management of the disease is difficult and economic and alternative solutions are needed. The medicinal plants compounds are known to have antimicrobial activity, and they could be an alternative choice in the C. cacaofunesta control. Considering this, this work aimed to verify the in vitro antifungal activity of aqueous and alcoholic solutions of Adiantum latifolium leaves on C. cacaofunesta. Plant material was collected at Atlantic Forest biome in cacao cultivation area in South of Bahia state. Aqueous and ethanolic solutions were made by boiling and maceration in 70% ethanol, respectively. After filtration, they were added to culture medium at 1, 5 and 10% dilution. A 7 mm disc colony of C. cacaofunesta was inoculated in the middle of the well containing Sabouraud dextrose agar (SDA) and the mycelial growth was observed. Controls consisted on SDA with sterile water or 70% ethanol at the same dilution of treatments, and Tebuconazole at 4 µg.mL-1. Neither aqueous nor ethanolic solutions inhibited the mycelial growth. However, aqueous solution presence induced a higher mycelial growth rate. Conversely, aqueous solution treatment induced mycelial growth. Tebuconazole showed important mycelial growth inhibition and it could be considered in C. cacaofunesta propagation control in areas where genetic selection or handling management still fail.(AU)

A espécie Ceratocystis cacaofunesta é o agente etiológico do mal-do-facão, patogenia caracterizada por danos irreversíveis no sistema vascular da planta. O controle da doença é difícil e a busca por soluções alternativas e econômicas é necessária. Sabe-se que os compostos das plantas medicinais possuem atividade microbiana e podem ser uma opção alternativa no controle de C. cacaofunesta. Baseado nisso, esse trabalho se propôs a verificar in vitro o potencial antifúngico das soluções aquosa e alcóolica de Adiantum latifolium sobre C. cacaofunesta. O material vegetal foi coletado no bioma Mata Atlântica em área de plantio de cacau, no sul da Bahia. Solução aquosa foi obtida por decocção e solução etanólica por maceração em etanol 70%. As soluções foram filtradas e adicionadas ao meio de cultura em diluições de 1, 5 e 10%. Inocularam-se fragmentos de 7 mm de colônia de C. cacaofunesta no centro do meio de cultura contendo ágar Sabouraud dextrose (ASD) e se observou o crescimento do disco micelial. Os controles consistiram em SDA com água estéril ou etanol a 70% na mesma diluição de tratamentos e o antifúngico Tebuconazol a 4 µg.mL-1. Nenhuma concentração das soluções aquosa e alcóolica inibiu o crescimento micelial. Entretanto, a presença de solução aquosa induziu maior crescimento micelial. O antifúngico Tebuconazol apresentou efeito redutor importante do crescimento micelial e pode ser uma alternativa no controle da propagação do C. cacaofunesta em locais onde a seleção genética e o manejo adequado de instrumentos no momento da poda apresentam falhas.(AU)

Adipose-derived stem cells (ADSC) in the viability of a random pattern dorsal skin flap in rats

PURPOSE: To evaluate the viability of random pattern dorsal skin flaps in rats after injection of adipose-derived stem cells (ADSC). METHODS: Thirty five adult male Wistar EPM rats (weight 250-300 g) were distributed, at random, in two groups. I- Control (flap elevation with injection of saline solution) with fifteen animals and II- Experimental (flap elevation with injection of ADSC ) with fifteen animal. The ADSC were isolated from others five adult male rats. A dorsal skin flap measuring 10x4 cm was raised and a plastic barrier was placed between the flap and its bed in both groups and the injection (cells or saline solution) were perfomed immediately after the surgery. The percentage of flap necrosis was measured on the seventh postoperative day. RESULTS: The ADSC were able to replicate in our culture conditions. We also induced their adipogenic, osteogenic and chondrogenic differentiation to verify their mesenchymal stem cells potentiality in vitro. The results were statistically significant showing that the ADSC decreased the area of necrosis (p<0.05). CONCLUSIONS: The cells demonstrated adipogenic, osteogenic and chondrogenic differentiation potential in vitro. The administration of adipose-derived stem cells was effective to increase the viability of the random random pattern dorsal skin flaps in rats. .

Evaluation of antimicrobial and antitumoral activity of Garcinia mangostana L. (mangosteen) grown in Southeast Brazil

PURPOSE: To characterize the anatomy of the fruit and leaf and the presence of phytocompounds. To evaluate the antitumor and antimicrobial activity of ethanolic extract of Garcinia mangostana L. (mangosteen) cultivated in southeastern Brazil. METHODS: Anatomical characterization and histochemical reactions were performed for structural identification and the presence of phytocompounds. Preparation of ethanolic extract of the fruit, leaf and resin of mangosteen. Culture B16-F10 melanoma cells for treatment with mangosteen ethanolic extract to determine cell viability by MTT and genotoxic effect by comet assay. Evaluation by antimicrobial activity against Staphylococcus aureus and Escherichia coli by agar diffusion test and by determination of Minimum Inhibitory Concentration (MIC). RESULTS: Our results showed many secretory canals in resin fruit and leaf; identifying lipids, starch, lignin and phenolic compounds. The leaf extract induced genotoxicity and apoptosis in B16-F10 cells, since the fragmentation of DNA in the comet assay. The ethanolic extract of mangosteen obtained in the resin, leaf and fruit showed antimicrobial activity against Staphylococcus aureus and Escherichia coli with a MIC at 0.1 mg/mL. CONCLUSION: In conclusion, we have demonstrated both antimicrobial and antitumor activity of ethanol extract of mangosteen emphasizing its therapeutic potential in infectious diseases and in cancer, such as melanoma. .

Development of experimental in vitro burn model

PURPOSE: To propose an experimental burn model in NIH-3T3 cell line. METHODS: Induction of thermal injury in cultures of mouse fibroblast - NIH-3T3- cell line and determination of cell viability by MTT and imunofluorescence. RESULTS: The heating of the Petri dish increased proportionally to the temperature of the base and the time of exposure to microwave. In this in vitro burn model, using the cell line NIH-3T3 was observed drastic cellular injury with significant changes in cell viability and activity. It showed drastically modified cell morphology with altered membrane, cytoskeleton and nucleus, and low cellularity compared to the control group. CONCLUSION: The burn model in vitro using the cell line NIH-3T3 was reproductive and efficient. This burn model was possible to determine significant changes in cell activity and decreased viability, with drastic change in morphology, cell lysis and death. .

Adipose-derived stem cells (ADSC) in the viability of random skin flap in rats

PURPOSE: To evaluate the effects of the adipose-derived stem cells (ADSC) in the viability of random skin flap in rats. METHODS: Thirty five adult male Wistar rats (weight 250-300 g) were used. ADSC were isolated from adult male rats (n=5). ADSC were separated, cultured and then analyzed. A dorsal skin flap measuring 10x4 cm was raised and a plastic barrier was placed between the flap and its bed. After the surgical procedure, the animals were randomized into two groups (n=15 each group), group control and group ADSC. In all groups the procedures were performed immediately after the surgery. The percentage of flap necrosis was measured on the seventh postoperative day. RESULTS: The ADSC were able to replicate in our culture conditions. We also induced their adipogenic, osteogenic and chondrogenic differentiation, verifying their mesenchymal stem cells potentiality in vitro. The results were statistically significant showing that the ADSC decreased the area of necrosis (p<0.05). CONCLUSION: The cells demonstrated adipogenic, osteogenic and chondrogenic differentiation potential in vitro. The administration of adipose-derived stem cells was effective to increase the viability of the random skin flaps in rats. .

Evaluation of antitumoral and antimicrobial activity of Morinda lcitrifolia L. grown in Southeast Brazil

PURPOSE: To evaluate the antitumor and antimicrobial activity of ethanolic extract of Morinda citrifolia L. fruit cultivated in southeastern Brazil. METHODS: Preparation ethanolic extract of the fruit of Morinda citrifolia L. Culture of melanoma cells B16-F10 for treatment with ethanolic extract of Morinda citrifolia L. fruit to determine cell viability by MTT and determination temporal effect of ethanolic extract fruit on the cell growth B16-F10 for 8 days. Evaluation of antimicrobial activity of ethanolic extract fruit against Staphylococcus aureus and Escherichia coli by determination of Minimum Inhibitory Concentration (MIC). RESULTS: The ethanolic extract of Morinda citrifolia L. fruit (10mg/mL) decreased cellular activity and inhibited 45% the rate of cell proliferation of B16-F10 melanoma treated during period studied. The ethanolic extract of Morinda citrifolia L. fruit demonstrated antimicrobial activity inhibiting the growth of both microorganisms studied. Staphylococcus aureus was less resistant to ethanolic extract of Morinda citrifolia L. fruit than Escherichia coli, 1 mg/mL and 10 mg/mL, respectively. CONCLUSION: What these results indicate that the ethanolic extract of the fruit of Morinda citrifolia L. showed antitumor activity with inhibition of viability and growth of B16-F10 cells and also showed antibacterial activity as induced inhibition of growth of Staphylococcus aureus and Escherichia coli. .

Characterization of human adipose-derived stem cells / Caracterização de células-tronco do tecido adiposo humano

PURPOSE: There is a growing scientific interest in the plasticity and therapeutic potential of adipose-derived stem cells (ASCs), which are multipotent and abundant in adipose tissue and can differentiate in vitro into multiple lineages, including adipocytes, chondrocytes, osteoblasts, neural cells, endothelial cells and cardiomyocytes. The aim of this study was to isolate, cultivate and identify ASCs. METHODS: Human adipose precursor cells were obtained from subcutaneous abdominal tissue. Recently dispersed cells were separated by density centrifugation gradient, cultured and then analyzed. RESULTS: Human ASCs were able to replicate in our culture conditions. The cells maintained their phenotypes throughout the studied period on different passages confirming they suitability for in vitro cultivation. We also induced their adipogenic, osteogenic and chondrogenic differentiation, verifying their mesenchymal stem cells potentiality in vitro. Flow cytometry results showed that these cells expressed CD73, CD90 and CD105, (mesenchymal stem-cells markers), contrasting with the lack of expression of CD16, CD34 and CD45 (hematopoietic cells markers). CONCLUSION: It was possible to isolate human adipose-derived stem cells by in vitro cultivation without adipogenic induction, maintaining their functional integrity and high proliferation levels. The cells demonstrated adipogenic, osteogenic and chondrogenic differentiation potential in vitro.

OBJETIVO: Há um interesse científico crescente na plasticidade e potencial terapêutico das células-tronco do tecido adiposo humano, células multipotentes e abundantes no tecido adiposo que podem se diferenciar in vitro em múltiplas linhagens celulares, incluindo adipócitos, condrócitos, osteoblastos, células neurais, endoteliais e cardiomiócitos. O objetivo deste estudo foi isolar, cultivar e identificar células-tronco do tecido adiposo humano. MÉTODOS: Células precursoras humanas do tecido adiposo foram obtidas de tecido abdominal subcutâneo. As células recém-dispersas foram separadas por gradiente de centrifugação por densidade, cultivadas e então analisadas. RESULTADOS: As células-tronco do tecido adiposo humano foram capazes de se replicar nas nossas condições de cultivo e mantiveram seu fenótipo em diferentes passagens durante o estudo, confirmando sua adequabilidade para cultivo in vitro. A diferenciação adipogênica, osteogênica e condrogênica também foi induzida, confirmando seu potencial de células-tronco mesenquimais in vitro. Os resultados de citometria de fluxo evidenciaram a expressão dos marcadores de células-tronco mesenquimais CD73, CD90 e CD105, contrastando com a falta de expressão dos marcadores de células hematopoiéticas CD16, CD34 e CD45. CONCLUSÃO: Foi possível isolar células-tronco do tecido adiposo humano por cultivo in vitro sem indução adipogênica, mantendo sua integridade funcional e altos níveis de proliferação. As células demonstraram potencial de diferenciação adipogênico, osteogênico e condrogênico in vitro.

Avaliação morfológica e dos mecanismos de mobilização de Ca2+ pela glicose e acetilcolina em células pancreáticas humanas / Morphologic evaluation and Ca2+ mobilization by glicose and acetylcholine in human pancreatic cells

OBJETIVOS: Avaliar a morfologia das organelas e do citoesqueleto em células pancreáticas humanas cultivadas e a mobilização de Ca2+ em resposta à glicose e ACh por medidas fluorimétricas. MATERIAL E MÉTODOS: As células foram semeadas em lamínulas, fixadas e marcadas com uma combinação de fluoróforos: o núcleo foi corado com DAPI e as mitocôndrias, com Mytotracker Red. Foram utilizados faloidina e anticorpos secundários conjugados com Alexa Fluor verde e vermelho fluorescentes (488 e 594) para identificar proteína actina F e receptor muscarínico tipo M3, respectivamente. Para estudar a mobilização de Ca2+, as células foram incubadas com fura-2/AM. RESULTADOS: As células pancreáticas humanas apresentaram morfologia preservada com grande quantidade de mitocôndrias. Na região de maior densidade celular, evidenciou-se as pseudo-ilhotas e os receptores muscarínicos M3. Por meio da elevação da [Ca2+]c, devido à ação da glicose e ACh, mostrou-se preservação da capacidade responsiva a esses estímulos e foi dependente de concentração desses agonistas. A glicose promoveu uma resposta sustentada e a ACh induziu uma resposta bifásica. CONCLUSÃO: As células pancreáticas humanas cultivadas conservaram sua morfologia. A mobilização de Ca2+ em resposta à glicose e a ACh confirma a sua funcionalidade. Os receptores muscarínicos M3 estão presentes nessas células.

AIMS: The proposal of this study was to analyze morphology of the organelles and cytoskeleton in human pancreatic cells cultured and the mobilization of the cytosolic calcium ([Ca2+]c) in response to glucose and ACh by fluorimetry method. MATERIAL AND METHODS: The cells were plated on glass coverslips, fixed and stained with a combination of fluorophores: the nuclei were stained with DAPI and mitochondria with Mytotracker Red. It was used phalloidin and the secondary antibodies Alexa Fluor conjugated green and red-fluorescent (488 and 594) to identify the protein cell actin F and type M3 muscarinic receptor respectively. The cells also were loaded with fura-2/AM to study Ca2+ mobilization. RESULTS: The human pancreatic cells show characteristics morphologically preserved with great amount of mitochondria. In region major cell density was evidenced pseudo-islets and type M3 muscarinic receptors. Through increase of [Ca2+]c due to action of glucose and ACh were shown that the cellsÆ capacity to respond to these stimuli were conserved. The elevation of the [Ca2+]c depended on concentration by glucose-induced promoting sustained phase and ACh-induced a biphasic response. CONCLUSION: The morphologic characteristics of human pancreatic cells cultured were preserved. The Ca2+ mobilization in response to glucose and ACh confirmed its functionality. The expression of the M3 muscarinic receptors in human pancreatic cell cultured was demonstrated.

Uma síntese sobre colifagos como indicadores de poluição fecal / Colifags as Indicators of Fecal Pollution - a Summary

Em 1917 alguns autores descreveram um fenômeno lítico não específico em placas de culturas bacterianas. Este princípio lítico levantou a hipótese de que se tratava de vírus patogênico para a bactéria que causava lise de seu hospedeiro enquanto se multiplicava. Este microorganismo foi denominado bacteriófago ou fago. Colifago é o nome genérico aplicado a bacteriófagos que atacam bactérias do grupo coliforme. Acredita-se que se os colifagos estejam presentes sempre que as bactérias coliformes fecais são isoladas. Atualmente, a detecção de níveis de bacteriófagos na água poluída por esgoto tem sido proposta por diversos autores como forma de avaliar o nível de contaminação dessa água. A descrição de uma correlação entre níveis de colifagos e coliformes tem estabelecido a importância do bacteriófago como um indicador de contaminação fecal. Apesar do índice de coliformes ser o critério microbiólogico utilizado para avaliar a qualidade da água e o risco de presença de bactérias patogênicas, questiona-se a sua utilidade para indicar a presença de vírus entéricos. A detecção de níveis de bacteriógagos na água tem sido então proposta como alternativa à avaliação da contaminação de diferentes tipos de água tanto por bactérias quanto por vírus enteropatogênicos.

Efeito radioprotetor do alfa-tocoferol em íleo de camundongos expostos a radiaçäo gama: modificaçöes morfológicas, bioquímicas e funcionais / Effect radioprotector of alpha tocopherol in ilium of rabbits exposts on gama radiation: morphological, biochemical and functional modifications

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