In vitro Evaluation of Anti-Inflammatory Activity of "Habanero" Chili Pepper (Capsicum chinense) Seeds Extracts Pretreated with Cellulase.

The aim of this study was to explore the effect of capsaicin and particular phenolic compounds profile from cellulase assisted extracts of Habanero (Capsicum chinense) chili pepper seeds (CPS) on the concentration of cytokines (IL-2, IL-6, TNF-α, IL-1ß) in murine macrophages (RAW 264.7) stimulated with lipopolysaccharides (LPS). Capsaicin was quantified by HPLC-DAD, and the phenolic profile was determined by UPLC-MS-QqQ. Anti-inflammatory activity was evaluated by Mouse Cytokine/Chemokine Magnetic Bead Panel 96-well plate assay. Among the 15 different phenolics found in CPS extracts obtained at 120 or 150 min of maceration with 2,500 UI/L at 30 ºC or 45 ºC in a 1:15 (w:v) proportion, the most abundant was vanillic acid (7.97-12.66 µg/g). The extract obtained at 30 ºC and 120 min, showed similar effects than the observed for synthetic anti-inflammatory drugs indomethacin and dexamethasone, and capsaicin standard. Beyond capsaicin, salicylic, protocatechuic and trans-cinnamic acids as well as vanillin in CPS extracts were correlated with the anti-inflammatory effect. On the other hand, capsaicin and chlorogenic acid contents were potential immunostimulants whose concentration varied depending on the cellulase treatment time.

Enzyme-assisted extraction of anti-inflammatory compounds from habanero chili pepper (Capsicum chinense) seeds.

Capsaicinoids are the main bioactive compounds extracted from chili pepper seeds (CPSs) but other bioactive compounds such as phenolic compounds may be found. Enzyme-assisted extraction (EAE) improves the extraction of bioactive compounds from fruits and seeds. The aim of this study was to establish the cellulase-assisted extraction conditions of capsaicinoids and phenolic compounds from Habanero CPSs (Capsicum chinense) and to evaluate the anti-inflammatory activity of the obtained extracts on murine macrophages. EAE was performed using different temperatures (T1 = 30°C, T2 = 45°C and T3 = 60°C), enzyme concentrations (E1 = 2,500 UI/L and E2 = 250 UI/L), and extraction time periods (0-150 min). Total phenolic compounds were quantified using the Folin-Ciocalteu assay, capsaicin (CAP) and dihydrocapsaicin (DHC) contents were evaluated by HPLC, and anti-inflammatory activity was performed with Griess assay on murine macrophage RAW 264.7 cell culture. The highest phenolic compound content (337.96 mg GAE/L) was achieved at 30°C, 2,500 UI/L, and 150 min of extraction. The highest CAP content (310.23 µg/ml) was obtained at 45°C with 250 UI/L for 150 min, while for DHC (167.72 µg/ml), the conditions were 60°C, 2,500 UI/L, and 120 min. The highest anti-inflammatory response was obtained when 60°C, E2, and 150 min were used for the extraction, and nitric oxide (NO) production was reduced to 22.56%. Based on the results obtained in this research, EAE allowed the recovery of compounds with anti-inflammatory activity from CPS using water as a solvent. There was a correlation between the extraction of CAP and DHC. But although a moderate direct correlation between the concentration of capsaicinoids and total phenolic compounds (TPCs) and an inverse correlation of the presence of the bioactive compounds (TPC, CAP, and DHC) with the NO synthesis, these were not statistically significant. We demonstrated that Habanero seeds are an important raw material to recover anti-inflammatory compounds beyond capsaicinoids using water in EAE.

Inhibition of miR31 and miR92a as Oncological Biomarkers in RKO Colon Cancer Cells Treated with Kaempferol-3-O-Glycoside Isolated from Black Bean.

MicroRNAs (miRNAs) are small molecules of 19-23 nucleotides of RNA that act as regulators of the expression of proteins in eukaryotic cells. Currently, the participation of miRNAs in the development of different types of cancer has been observed. To evaluate the inhibitory effect of kaempferol-3-O-glycoside on the expression of oncological biomarkers, miR31 and miR92a in a colon cancer cell line (RKO) were analyzed. Cells were cultured and treated with 1 mM kaempferol-3-O-glycoside isolated from black bean. Expression levels of miR31 and miR92a were evaluated by real-time PCR using TaqMan probes; in addition, two oncogenes (KRAS and c-MYC) and two tumor suppressors (AMP-activated protein kinase [AMPK] and adenomatous tumors of polyposis coli [APC]) were quantified to validate the biological effects; normalization of expression levels were carried out by 2-ΔΔCt. Results were analyzed by one-way ANOVA. The expression levels of miR31, miR92a, KRAS oncogene, and the c-MYC transcription factor were subexpressed upon 72 h post-treatment with kaempferol-3-O-glycoside compared with the control without treatment (P < .05); in contrast, the tumor suppressor genes AMPK (∼4.85, P = .005) and APC (∼2.71, P = .066) tumor suppressors genes were overexpressed. Our results showed the inhibitory effect of isolated black bean flavonoid kaempferol-3-O-glycoside on cancer biomarkers: miR31 and miR92a; based on our results, this flavonoid may have interesting nutritional, therapeutic, and/or prophylactic applications to combat colon cancer.

Isorhamnetin glycoside isolated from Opuntia ficus-indica (L.) MilI induces apoptosis in human colon cancer cells through mitochondrial damage.

This work aimed to evaluate the mechanisms involved in the apoptosis induction of isorhamnetin-3-O-glucosyl-pentoside (IGP) in metastatic human colon cancer cells (HT-29). To achieve this, we assessed phosphatidylserine (PS) exposure, cell membrane disruption, chromatin condensation, cell cycle alterations, mitochondrial damage, ROS production, and caspase-dependence on cell death. Our results showed that IGP induced cell death on HT-29 cells through PS exposure (48%) and membrane permeabilization (30%) as well as nuclear condensation (54%) compared with control cells. Moreover, IGP treatment induced cell cycle arrest in G2/M phase. Bax/Bcl-2 ratio increased and the loss of mitochondrial membrane potential (63%) was observed in IGP-treated cells. Finally, as apoptosis is a caspase-dependent cell death mechanism, we used a pancaspase-inhibitor (Q-VD-OPh) to demonstrate that the cell death induced by IGP was caspase-dependent. Overall these results indicated that IGP induced apoptosis through caspase-dependent mitochondrial damage in HT-29 colon cancer cells.

Nutraceutical Potential of Five Mexican Brown Seaweeds.

In search of pharmaceutically active products to control type 2 diabetes, five brown seaweeds (Silvetia compressa, Cystoseira osmundacea, Ecklonia arborea, Pterygophora californica, and Egregia menziesii) from the Northwest Mexican Pacific coast were investigated. Proximate composition and total polyphenol content (TPC) as phloroglucinol equivalents (PGE) were determined for the five seaweed powders and their respective hydroethanolic (1 : 1) extracts. Extracts were screened for their radical scavenging activity (DPPH and ORAC) and glycosidase inhibitory activity. HPLC-DAD, HPLC-MS-TOF, and ATR-FT-IR methodologies were used to identify the most abundant phlorotannins and sulfated polysaccharides in the extracts. Hydroethanolic extracts contained minerals (17 to 59% of the dry matter), proteins (4 to 9%), ethanol-insoluble polysaccharides (5.4 to 53%), nitrogen-free extract (NFE) (24.4 to 70.1%), lipids (5 to 12%), and TPC (2.6 to 47.7 g PGE per 100 g dry extract). S. compressa and E. arborea dry extracts presented the lowest ash content (26 and 17%, respectively) and had some of the highest phenolic (47.7 and 15.2 g PGE per 100 g extract), NFE (57.3 and 70.1%), and soluble polysaccharide (19.7 and 53%) contents. S. compressa and E. arborea extracts had the highest antioxidant activity (IC50 DPPH 1.7 and 3.7 mg mL-1; ORAC 0.817 and 0.801 mmol Trolox equivalent/g extract) and the highest α-amylase and α-glucosidase inhibitory capacities (IC50 940 and 1152 µg mL-1 against α-amylase and 194 and 647 µg mL-1 against α-glucosidase). The most abundant phlorotannins identified in the extracts were phloretol, fucophloroethol, and two- and three-phloroglucinol unit (PGU) phlorotannins. Laminarin, fucoidan, and alginate were among the sulfated polysaccharides identified in the extracts. The bioactivities of S. compressa and E. arborea extracts were mainly related with their contents of three PGU phlorotannins and sulfated polysaccharides (e.g., fucoidan, laminarin, and alginate). These results suggest S. compressa and E. arborea are potential candidates for food products and nutraceutical and pharmaceutical preparations, and as additives for diabetes management.

Peptides and isoflavones in gastrointestinal digests contribute to the anti-inflammatory potential of cooked or germinated desi and kabuli chickpea (Cicer arietinum L.).

It is largely unknown how processing affects bioactive potential of chickpea proteins to prevent bowel inflammatory diseases. The aim was to investigate the anti-inflammatory activity of protein concentrates from germinated and cooked chickpeas (GC and CC, respectively) and its relationship with protein and isoflavone composition before and after in vitro gastrointestinal digestion and absorption. Anti-inflammatory activity of GC digests was almost 2-fold higher than CC digests (p < 0.05), which was associated to greater content of peptides, formononetin and biochanin A (p < 0.05). Anti-inflammatory activity of phenolic fraction in digests was 7-fold higher than the protein fraction (p < 0.05). The most active peptide fraction from GC digest (IC50 = 93 µg/mL) contained a total of 24 peptides derived from legumin and vicilin. In conclusion, this study stands out the potential of germinated chickpea proteins concentrates to exert anti-inflammatory effects in the lower gut which may contribute to the prevention of bowel inflammatory diseases.

Smilax aristolochiifolia Root Extract and Its Compounds Chlorogenic Acid and Astilbin Inhibit the Activity of α-Amylase and α-Glucosidase Enzymes.

Regulating activities of α-amylase and α-glucosidase through the use of specific inhibitors is a main strategy for controlling type 2 diabetes. Smilax aristolochiifolia root decoctions are traditionally used in Mexico as hypoglycemic and for weight loss, but the active principles and mechanisms underlying such putative metabolic effects are yet unknown. Here, we isolated the major bioactive compounds from a hydroethanolic extract of S. aristolochiifolia root by fast centrifugal partition chromatography and evaluated their effects against pancreatic α-amylase and yeast α-glucosidase. A chlorogenic acid-rich fraction (CAF) inhibited α-amylase activity with an IC50 value of 59.28 µg/mL in an uncompetitive manner and α-glucosidase activity with an IC50 value of 9.27 µg/mL in a noncompetitive mode. Also, an astilbin-rich fraction (ABF) inhibited α-glucosidase activity with an IC50 value of 12.30 µg/mL, in a noncompetitive manner. CAF inhibition α-amylase was as active as acarbose while both CAF and ABF were 50-fold more potent inhibitors of α-glucosidase than acarbose. The molecular docking results of chlorogenic acid and astilbin with α-amylase and α-glucosidase enzymes correlated with the inhibition mechanisms suggested by enzymatic assays. Our results prove that S. aristolochiifolia roots contain chlorogenic acid and astilbin, which inhibit carbohydrates-hydrolyzing enzymes, suggesting a new mechanism for the hypoglycemic effect reported for this plant.

Poly(3-hydroxybutyrate) graft copolymer dense membranes for human mesenchymal stem cell growth

Background: The use of novel materials as an artificial extracellular matrix for stem cell growth is a current strategy of increasing interest for regenerative medicine. Here, we prepare thermal-remolded membrane scaffolds from poly(3-hydroxybutyrate) grafted with 2-amino-ethyl methacrylate hydrochloride. However, it is unclear whether these membranes are useful for tissue engineering. Results: The mechanical properties, tribology, and morphology of the dense membranes were assessed. The results show that tensile strain at break and roughness of the compressed membrane decrease with increasing graft degree. Moreover, graft copolymer membranes showed lower resistance to scratching, greater degree of swelling and higher brittleness than un-grafted P(3HB) films. Thus, it effectively supports the growth of dermal fibroblast, as demonstrated by epifluorescence microscopy. Conclusions: It is concluded that the developed membrane can be properly used in is the restoration of skin tissue. How to cite: González-Torres M, Sánchez-Sánchez R, Solís-Rosales SG, et al. Poly(3-hydroxybutyrate) graft copolymer dense membranes for human mesenchymal stem cell growth.

Effect of Agave americana and Agave salmiana Ripeness on Saponin Content from Aguamiel (Agave Sap).

Steroidal saponins have shown beneficial health effects. Agave spp. leaves and rhizomes are sources of these compounds, but their presence has not been reported in the aguamiel. Aguamiel is the sweet edible sap from mature agave, and its quality is influenced by the plant ripening stage. The purpose of this research was to identify and quantitate saponins in aguamiel from Agave americana and Agave salmiana at two ripening stages. Saponins and sapogenins were identified with HPLC/ESI-MS/TOF and quantitated with HPLC/ELSD. Results proved the presence of saponins derived from kammogenin, manogenin, gentrogenin, and hecogenin. The saponin content in aguamiel from immature A. salmiana was 2-fold higher (478.3 protodioscin equivalents (PE) µg/g aguamiel (DM)) compared with A. americana (179.0 PE µg/g aguamiel (DM)). In both species, saponin content decreased when plants reached sexual maturity. This should be considered before evaluating the effects of Agave spp. as a source of bioactive saponins.

Isolation and structure elucidation of avocado seed (Persea americana) lipid derivatives that inhibit Clostridium sporogenes endospore germination.

Avocado fruit extracts are known to exhibit antimicrobial properties. However, the effects on bacterial endospores and the identity of antimicrobial compounds have not been fully elucidated. In this study, avocado seed extracts were tested against Clostridium sporogenes vegetative cells and active endospores. Bioassay-guided purification of a crude extract based on inhibitory properties linked antimicrobial action to six lipid derivatives from the family of acetogenin compounds. Two new structures and four compounds known to exist in nature were identified as responsible for the activity. Structurally, most potent molecules shared features of an acetyl moiety and a trans-enone group. All extracts produced inhibition zones on vegetative cells and active endospores. Minimum inhibitory concentrations (MIC) of isolated molecules ranged from 7.8 to 15.6 µg/mL, and bactericidal effects were observed for an enriched fraction at 19.5 µg/mL. Identified molecules showed potential as natural alternatives to additives and antibiotics used by the food and pharmaceutical industries to inhibit Gram-positive spore-forming bacteria.