Cytotoxicity Screening of Sterculia striata A.St.-Hil. & Naudin (Chichá) and Arachis hypogaea L. (Peanut) and Comparative Chemical Profiles Before and After in Vitro Digestion.

This study traced the cytotoxicity, antioxidant activity, and phytochemical profile before and after in vitro digestion of nuts from Sterculia striata A. St.-Hil. & Naudin (Malvaceae) (chichá or monkey's peanut), a native plant from Brazil, in comparison with Arachis hypogaea L. (peanut). The antioxidant activity in the 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and Ferric Reducing Antioxidant Power Assay (FRAP) assays was lower in chichá when compared with peanuts, corroborating the lower concentration of polyphenols. None of the samples studied showed significant cytotoxicity in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromideDAD: diode-array detection (MTT) assays. In vitro digestion altered the phytochemical profile in both plants, increasing the concentration of rutin in fresh and roasted chichá but only in raw peanuts. In roasted peanuts, rutin was converted into quercetin. Chichá nuts have been used by the local population for centuries, and the identification of their bioactive components can be useful to promote their benefits as a functional food.

Planar bioluminescent cytotoxicity assay via genetically modified adherent human reporter cell lines, applied to authenticity screening of Saussurea costus root.

The hyphenation of high-performance thin-layer chromatography to effect-directed assays is a very straightforward way to detect individual bioactive zones, and at the same time, to investigate several samples simultaneously. The combination of the separation technique with adherent human cells applied on the same surface was recently shown to be possible. Since on-surface adherent cell assays are in their infancy, a planar bioluminescent cytotoxicity assay was developed to expand the possibilities. Human embryonic kidney (HEK) 293 or HeLa (cervical carcinoma) cells were chosen because of their fast growth rates and high rates of successful transfection, being suitable for the generation of genetically modified reporter cells. For the first time, HeLa cells were visualized on the wettable reversed phase plate surface using digital microscopy. For the generation of bioluminescent reporter cells, vectors for the expression of three luciferase enzymes of various origins were tested. The genetically modified HEK 293T-CMV-ELuc cells were the best suitable for the new planar cytotoxicity assay due to the faster growth rate, robustness, and stronger bioluminescence signal. The stable expression of luciferase under the control of a strong constitutive promoter allowed the cells to be used for the determination of the cytotoxicity of Saussurea costus root samples obtained from the market and to assess the authenticity of these samples. Any cytotoxic zone was detected as a dark zone inhibiting the cell bioluminescence. Five replicates of the dose-response curve confirmed the good assay performance and the cytotoxicity of a zone, which was assigned to costunolide and dehydrocostus lactone. By this, the proof-of-principle of the new planar bioluminescent cytotoxicity assay, which does not require expensive licensing, was successful.

PAF signaling plays a role in obesity-induced adipose tissue remodeling.

BACKGROUND/OBJECTIVES: Platelet-activating factor receptor (PAFR) activation controls adipose tissue (AT) expansion in animal models. Our objective was twofold: (i) to check whether PAFR signaling is involved in human obesity and (ii) investigate the PAF pathway role in hematopoietic or non-hematopoietic cells to control adipocyte size. MATERIALS/SUBJECTS AND METHODS: Clinical parameters and adipose tissue gene expression were evaluated in subjects with obesity. Bone marrow (BM) transplantation from wild-type (WT) or PAFR-/- mice was performed to obtain chimeric PAFR-deficient mice predominantly in hematopoietic or non-hematopoietic-derived cells. A high carbohydrate diet (HC) was used to induce AT remodeling and evaluate in which cell compartment PAFR signaling modulates it. Also, 3T3-L1 cells were treated with PAF to evaluate fat accumulation and the expression of genes related to it. RESULTS: PAFR expression in omental AT from humans with obesity was negatively correlated to different corpulence parameters and more expressed in the stromal vascular fraction than adipocytes. Total PAFR-/- increased adiposity compared with WT independent of diet-induced obesity. Differently, WT mice receiving PAFR-/--BM exhibited similar adiposity gain as WT chimeras. PAFR-/- mice receiving WT-BM showed comparable augmentation in adiposity as total PAFR-/- mice, demonstrating that PAFR signaling modulates adipose tissue expansion through non-hematopoietic cells. Indeed, the PAF treatment in 3T3-L1 adipocytes reduced fat accumulation and expression of adipogenic genes. CONCLUSIONS: Therefore, decreased PAFR signaling may favor an AT accumulation in humans and animal models. Importantly, PAFR signaling, mainly in non-hematopoietic cells, especially in adipocytes, appears to play a significant role in regulating diet-induced AT expansion.

Evidence of traditionality of Brazilian medicinal plants: The case studies of Stryphnodendron adstringens (Mart.) Coville (barbatimão) barks and Copaifera spp. (copaíba) oleoresin in wound healing.

ETHNOPHARMACOLOGICAL RELEVANCE: The World Health Organization (WHO) recognizes the potential of plants used in secular traditional medicine and considers this an important source of evidence to assess their effectiveness and safety. Brazil is rich in biodiversity and traditional uses based on the Amerindian culture. However, many processes started with the arrival of the Portuguese in the year 1500. The successive economic cycles, for example, led to destruction of native vegetation and an intense cultural erosion. As a consequence, the information about the use of plants in the past centuries are dispersed and without interpretation. In this study a methodology to evidence the traditionality of Brazilian plants was demonstrated using data about barbatimão barks (Stryphnodendron adstringens (Mart.) Coville - Fabaceae) and Copaiba oleoresin (Copaifera spp. - Fabaceae) in wound healing, was established. MATERIAL AND METHODS: Data about use of the plants were recovered from bibliography published between 1576 and 2011. The books (101) were classified using weights, considering the date of publication and the source of Information. Older books that describe primary information received weight 10, while books written more recently and with secondary information received weight 0.4. A score for each category of medicinal use was calculated based on the books weights and the frequency of citation. A review about the current use of both plants was also performed from ethnobotanical studies published in journals. RESULTS AND DISCUSSION: The traditional secular use of barks of barbatimão and oleoresin of copaiba to treat wounds was confirmed based on the historic bibliographic research. The most frequent use of barbatimão in a timeline of 500 years of Brazil's history, was as astringent, whereas for copaíba was as healing of skin and mucosal lesions. The continuous and current use of these plants to treat wounds, confirmed by recent ethnobotanical studies, is an indicative of the resilience of these remedies and their effectiveness. CONCLUSION: The use of preparations containing barbatimão barks and copaiba oleoresin can be considered effective in the treatment of wounds. Nonetheless, it is necessary to improve the quality of the formulas as established by WHO.

The interplay between Araçatuba virus and host signaling pathways: role of PI3K/Akt in viral replication.

In this study, we describe the interaction between Araçatuba virus (ARAV), a naturally occurring Brazilian vaccinia virus isolated from an outbreak at a dairy farm, and the host cell's signal transduction pathways. Even though ARAV infection led to phosphorylation of MAPKs MEK/ERK, JNK, and p38MAPK, genetic or pharmacological inhibition of these pathways had no impact on viral replication. We also provide evidence that ARAV stimulated the phosphorylation of Akt (PKB) at serine 473 (S473-P), a signaling event that is required for full activation of Akt during the infectious cycle. Furthermore, pharmacological inhibition of PI3K (LY294002) abrogated ARAV-induced Akt activation (S473-P) and affected early and late viral gene expression, which was followed by a decrease in virus yield (~1 log). Taken together, our data shed some light onto the biological differences between ARAV and vaccinia virus strain WR (VACV-WR), which could contribute, at least in part, to the low-virulence phenotype displayed by ARAV. Thus, while the requirement for the PI3K/Akt pathway for successful ARAV replication is also shared with VACV-WR and cowpox virus strain BR (CPXV-BR), ARAV showed a lower replicative capacity, as well as a smaller plaque-size phenotype after infection of A31 cells when compared to VACV-WR.