Identificación de fosforilaciones CaMKII-dependientes en mitocondrias aisladas de corazón de rata

La participación de CaMKII en la apoptosis ha sido demostrada hace varios años. En nuestro laboratorio hemos descrito que la muerte por apoptosis/necrosis inducida por isquemia/reperfusión (IR) es mediada por CaMKII a nivel del retículo sarcoplasmático (RS), y que además, en este proceso participan las mitocondrias. Aún no se sabe si la fosforilación dependiente de CaMKII de proteínas mitocondriales es responsable de parte del este efecto deletéreo en la IR. La hipótesis es que el aumento del Ca2+ citosólico y posterior activación de CaMKII en la IR, podría fosforilar proteínas no sólo del RS (ya demostrado) sino además de las mitocondrias, por un aumento en la concentración local de Ca2+ y transferencia en el microdominio mitocondrias-RS. Esto permitiría una activación de CaMKII localmente y la fosforilación de proteínas, afectando la función mitocondrial y contribuyendo al efecto deletéreo de la CaMKII en la IR.Facultad de Ciencias Médica

Technological development of hydroalcoholic extractive solutions from Calendula officinalis L. flowers by using factorial design

L., pelo processo de maceração (MAC) e de turbo-extração (TURB), empregando planejamento fatorial 32 (dois fatores e três níveis), para avaliar a influência do tempo de extração (MAC = 5, 10 e 15 dias; TURB = 5, 15 e 25 min) e da relação planta:solvente (2,5, 7,5 e 12,5%, m/V) sobre o teor de flavonóides totais e resíduo seco nas soluções extrativas. Para o processo de maceração, a relação planta: solvente de 12,5% e o tempo de 15 dias resultaram nas melhores condições para alcançar um maior teor de flavonóides totais, em relação à turbo-extração que obteve o mesmo desempenho em 5 min. Para ambos os processos, o resíduo seco foi proporcional à relação planta:solvente.The work aimed at the development of extractive solutions from the flowers of Calendula officinalis, through maceration (MAC) and turbo-extraction (TURB) processes, employing the 32 (two factors and three levels) factorial design, in order to evaluate the influence of the time (MAC = 5, 10 and 15 days; TURB = 5, 15 and 25 min) and of the plant: solvent ratio (2.5, 7.5 and 12.5%, w/v) on the flavonoid content and the dry residue yields in the extractive solutions. For the maceration process, the 12.5% (w/v) plant: solvent ratio over 15 days were the best conditions to achieve the highest flavonoid content, in the turbo-extraction process the same performance was achieved in 5 min. For both processes the dry residue was proportional to the plant: solvent ratio.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Anti-inflammatory, antiallodynic effects and quantitative analysis of gallic acid in spray dried powders from Phyllanthus niruri leaves, stems, roots and whole plant

The anti-inflammatory and antiallodynic effects of spray dried powders starting from leaves, stems, roots, the mixture of leaves and stems, as well as the whole plant aqueous solutions of Phyllanthus niruri L., Phyllanthaceae, were assessed. Gallic acid, used as chemical marker, was quantified by HPLC in the spray dried powders. Carrageenan-induced inflammatory and allodynic responses in the mouse paw were used as pharmacological models. Quantitative and qualitative differences among chemical composition of different herb parts were observed. The oral administration of leaves or leaves plus stems spray dried powders (100 mg/kg) significantly inhibited the carrageenan-induced allodynic effect (42±5 and 54±3%, respectively). Additionally, the spray dried powders of leaves significantly reduced carrageenan-induced paw oedema (35±6%). The spray dried powders of roots, stems, or the mixture of leaves, stems and roots (100 mg/kg, p.o.) did not exhibit antiallodynic or antioedematogenic effect in the same model. In conclusion, differences in the chemical composition of spray dried powders from P. niruri are reflected in their in vivo pharmacological actions. Despite of a direct relationship of anti-inflammatory and antiallodynic effects with the gallic acid content had been observed, especially in the spray dried powders of leaves, the use of spray dried powders of leaves plus stems showed to be more effective, suggesting a synergic effect between their constituents

Simultaneous determination of epicatechin and procyanidin A2 markers in Litchi chinensis leaves by high-performance liquid chromatography

The fruits of Litchi chinensis Sonn., Sapindaceae, are renowned for their biological activities. However, their leaves are poorly explored, although they represent an important source of vegetable raw material with biological properties as antioxidant, anti-inflammatory and antinociceptive. An HPLC method was developed and validated for the simultaneous quantification of epicatechin and procyanidin A2 in the leaf hydroethanolic extract of L. chinensis. The markers and other unidentified components were separated on a Luna Phenomenex C18 column (250 mm × 4.6 mm, 5 μm) with mobile phase composed of acetonitrile: water pH 3.0 (with sulfuric acid), in a gradient run; at 1.0 ml min−1, 30 °C and 278 nm for detection. The method was linear over an epicatechin and procyanidin A2 concentration range of 10–100 μg ml−1. The Limit of Quantification for epicatechin and procyanidin A2 were 1.7 and 2 μg ml−1, respectively. The Relative Standard Deviation (%) values for markers (intra- and inter-day precision studies) were <4.0% and the accuracy was 100 ± 5%. The method was applied to ten samples collected in the state of Santa Catarina (Brazil), which showed 14.8–44.5 and 44.8–69.6 mg g−1 of epicatechin and procyanidin A2, respectively. The proposed method could be a valuable tool for quality assessment of L. chinenis leaves as well as their herbal derivatives. Keywords: Analytical validation, Epicatechin, Leaf extract, LC-UV method, Litchi chinensis, Procyanidin A

Chemical and technological evaluation of the Phyllanthus niruri aerial parts as a function of cultivation and harvesting conditions

Tea from Phyllanthus niruri L., Phyllanthaceae, aerial parts is commonly used by Brazilian folk medicine for its benefits on the treatment of genitourinary disorders, for what the polyphenolic compounds are mainly responsible. The yield of such compounds may be influenced by several variables related with the plant growth. The effects of planting periods and harvesting conditions are investigated in this work, including four different seasons. The cultivation was characterized by dry mass yield of aerial parts, and the effect of pruning was analyzed. Leaves, stems and their mixtures were analyzed after drying and milling. Loss on drying and water soluble extractives were used as physical parameters for quality control. Flavonoid content and gallic acid were chosen as chemical markers for this work. The spectrophotometric trial based on the aluminum chloride complexes was applied to evaluate the total flavonoids content. Gallic acid contents were measured from the water extractive solutions by high-performance liquid chromatography. The pruning caused a positive influence on the amount of leaves and stems. The highest flavonoids and gallic acid contents were found in the leaves, which were developed over the summer and the winter, respectively, both from the second harvesting (after pruning). Chomatographic profile by HPLC was characterizes by the presence of gallic acid and two other major peaks (not identified substances), which relation was peculiar to each aerial part. In conclusion, these results suggest that even under less favorable climatic conditions, in winter, the pruning seems to cause a strong influence over the P. niruri polyphenolics production. Indeed, the total flavonoids content, as well as the HPLC profile, can be used as indicative parameters of the ratio of leaves and stem in the vegetal raw material