Characterization of Phenolic Compounds and Antioxidant and Anti-inflammatory Activities from Mamuyo (Styrax ramirezii Greenm.) Fruit.

Extracts of Styrax ramirezii Greenm., a fruit traditionally valued for health and wellness in Mexico, were analyzed phytochemically and evaluated for antioxidant and anti-inflammatory activity. Six norneolignans were identified by HPLC-TOF-MS, and the two major compounds were isolated for further evaluation. The effects of the isolated norneolignans, egonol and homoegonol, on lipopolysaccharide (LPS)-induced nitric oxide (NO) production, reactive oxygen species (ROS) production, and biomarkers of inflammation were evaluated. Of the tested compounds, egonol potently inhibited the production of NO and also significantly reduced the release of ROS. Consistent with these observations, the mRNA expression levels of inducible nitric oxide synthase (iNOS) (0.668 ± 0.108), cyclooxygenase-2 (COX-2) (0.553 ± 0.007), interleukin-1ß (IL-1ß) (0.093 ± 0.005), and interleukin-6 (IL-6) (0.298 ± 0.076) were reduced by egonol. The activity for both egonol and homoegonol increased in a concentration-dependent manner. These results suggest the potential of S. ramirezii Greenm. fruit to contribute to a healthy diet, rich in antioxidant and anti-inflammatory compounds.

Inhibition of pro-inflammatory responses and antioxidant capacity of Mexican blackberry (Rubus spp.) extracts.

Total polyphenolic and anthocyanin- and proanthocyanidin-rich fractions from wild blackberry genotypes (WB-3, WB-7, WB-10, and WB-11), a domesticated noncommercial breeding line (UM-601), and a commercial cultivar (Tupy) were evaluated for inhibition of pro-inflammatory responses [nitric oxide (NO) production, inducible nitric oxide synthase (iNOS) expression, cyclooxygenase-2 (COX-2) expression, and prostaglandin E2 (PGE2)] in RAW 264.7 macrophages stimulated by lipopolysaccharide (LPS). At 50 microM [cyanidin-3-O-glucoside (C3G) or catechin equivalent], most fractions significantly (P<0.05) inhibited all markers. The anthocyanin-rich fraction from WB-10 and the proanthocyanidin-rich fraction from UM-601 exhibited the highest NO inhibitory activities (IC50=16.1 and 15.1 microM, respectively). Proanthocyanidin-rich fractions from the wild WB-10 showed the highest inhibition of iNOS expression (IC50=8.3 microM). Polyphenolic-rich fractions from WB-7 and UM-601 were potent inhibitors of COX-2 expression (IC50=19.1 and 19.3 microM C3G equivalent, respectively). For most of the extracts, antioxidant capacity was significantly correlated with NO inhibition. Wild genotypes of Mexican blackberries, as rich sources of polyphenolics that have both antioxidant and anti-inflammatory properties, showed particular promise for inclusion in plant improvement programs designed to develop new varieties with nutraceutical potential.

Characterization of anthocyanins and proanthocyanidins in wild and domesticated Mexican blackberries (Rubus spp.).

This study was designed to characterize and compare wild, commercial, and noncommercial cultivated blackberry genotypes grown in Michoacan, Mexico. Six genotypes, including WB-3, WB-7, WB-10, and WB-11 (all wild blackberry types), Tupy (a commercial cultivar), and UM-601 (a cultivated breeding line), were selected and profiled for anthocyanins and proanthocyanidins by separating extracts over Amberlite XAD-7 resin and Sephadex LH-20 columns. Subsequent high-performance liquid chromatography (HPLC) and liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) analyses revealed that the major anthocyanin for all genotypes was cyanidin 3-O-glucoside. The proanthocyanidins (condensed tannins) were present in mono- to hexamer forms. Also, hydrolyzable tannins, ellagitannins, were characterized in the blackberry fruits. The average anthocyanin concentration in Sephadex LH-20 fractions was 49.2 mg/g in the commercial cultivar Tupy, while in the wild genotypes and the breeding line, the range was 361.3-494.9 mg/g (cyanidin 3-O-glucoside equivalent). The proanthocyanidin concentration varied widely among wild genotypes (417.5-1343.6 mg/g, catechin equivalent). This study demonstrated that the use of Amberlite XAD-7 followed by Sephadex LH-20 chromatography, with subsequent HPLC and LC-ESI-MS analyses, was able to effectively separate and characterize the diverse polyphenolics in blackberry genotypes. These results suggest that recommendations for dietary intake of blackberries for human health benefits need to take into account the source, because of the wide inherent variation in bioactive polyphenolic content in different blackberry genotypes.

Carotenogenic gene expression and ultrastructural changes during development in marigold.

Marigold (Tagetes erecta) flowers are a good source of carotenoids and can be used as a model studies on pigmentation during flower development. They show different levels of pigmentation caused by lutein. Here we describe the expression of several genes in the carotenoid biosynthetic pathway: phytoene synthase (Psy), phytoene desaturase (Pds), lycopene beta-cyclase (Lcy-b) and lycopene epsilon-cyclase (Lcy-e). cDNA inserts from isolated clones were 1376-1916bp long. The predicted amino acid sequences showed from 66 to 100% homology with other reported sequences (NCBI gene bank). Northern blot analyses of three varieties of marigold showed that most gene transcripts were expressed during flower development. The ultrastructural changes that occurred during plastid differentiation in flower morphogenesis were analyzed, and pigment accumulation among varieties was evaluated. The pigment deposition in specific structures (lipidic vesicles) during flower development was demonstrated.