Stimulant Paste Preparation and Bark Streak Tapping Technique for Pine Oleoresin Extraction.

Tapping technique comprises the extraction of pine oleoresin, a non-wood forest product consisting of a complex mixture of mono, sesqui, and diterpenes biosynthesized and exuded as a defense response to wounding. Oleoresin is used to produce gum rosin, turpentine, and their multiple derivatives. Oleoresin yield and quality are objects of interest in pine tree biotechnology, both in terms of environmental and genetic control. Monitoring these parameters in individual trees grown in the field provides a means to examine the control of terpene production in resin canals, as well as the identification of genetic-based differences in resinosis. A typical method of tapping involves the removal of bark and application of a chemical stimulant on the wounded area. Here we describe the methods for preparing the resin-stimulant paste with different adjuvants, as well as the bark streaking process in adult pine trees.

A Modified Protocol for High-Quality RNA Extraction from Oleoresin-Producing Adult Pines.

RNA extraction resulting in good yields and quality is a fundamental step for the analyses of transcriptomes through high-throughput sequencing technologies, microarray, and also northern blots, RT-PCR, and RTqPCR. Even though many specific protocols designed for plants with high content of secondary metabolites have been developed, these are often expensive, time consuming, and not suitable for a wide range of tissues. Here we present a modification of the method previously described using the commercially available Concert™ Plant RNA Reagent (Invitrogen) buffer for field-grown adult pine trees with high oleoresin content.

Box-Behnken factorial design to obtain a phenolic-rich extract from the aerial parts of Chelidonium majus L.

A Box-Behnken design (BBD) was developed to study the influence of four parameters (X1: % methanol; X2: extraction time; X3: extraction temperature; X4: solid/solvent ratio) on two responses, namely extraction yield and phenolics content of the aerial parts of Chelidonium majus L. The model presented a good fit to the experimental results for the extraction yield, being significantly influenced by X1 and X4. On the other hand a parameter reduction was necessary to run the model for phenolics content, showing that only X1 and X2 had great influence on the response. Two best extraction conditions were defined: X1=76.8% MeOH, X2=150.0 min, X3=60.0°C and X4=1:100 and X1=69.2%, X2=150 min, X3=42.5°C and X4=1:100. Moreover, the HPLC-DAD-ESI/MS(n) analysis conducted with the center point sample revealed the presence of 15 alkaloids and 15 phenolic compounds, from which the 9 flavonoids and 3 hydroxycinnamic acids are described for the first time. Only phenolic compounds were quantified by a validated HPLC-DAD method, the pair quercetin-3-O-rutinoside+quercetin-3-O-glucoside dominating all the 29 extracts. This study is of great importance for future works that seek to apply the phenolic profile to the quality control of C. majus samples.

A doença como crise na biografia do homem: um enfoque gastroenterológico / Disease as crisis in the biography of the man: a gastroenterologic approach

Os autores estudam o problema da doença como crise na biografia do homem, analisando-a do ponto-de-vista gastroenterológico. Apreciam a doença como geradora de uma crise e, inversamente, a crise como geradora de uma doença. Avaliam criticamente, em particular, o problema da úlcera péptica e da enterocolopatia funcional