Phenylethanoid Glycoside-Enriched Extract Prepared from Clerodendrum chinense Leaf Inhibits A549 Lung Cancer Cell Migration and Apoptosis Induction through Enhancing ROS Production.

This study aims to investigate the antioxidant and anti-cancer activities of Clerodendrum chinense leaf ethanolic extract. The phenylethanoid glycoside-enriched extract, namely verbascoside and isoverbascoside, was determined in the ethanolic C. chinense leaf extract using the validated HPLC method. The ethanolic extract showed DPPH and ABTS free radical scavenging activities with the IC50 values of 334.2 ± 45.48 µg/mL and 1012.77 ± 61.86 µg/mL, respectively, and a FRAP value of 88.73 ± 4.59 to 2480.81 ± 0.00 µM. C. chinense leaf extract exhibited anti-proliferative activity against A549 lung cancer cells in a dose- and time-dependent manner, with the IC50 value of 340.63 ± 89.43, 210.60 ± 81.74, and 107.08 ± 28.90 µg/mL after treatment for 24, 48, and 72 h, respectively. The IC50 values of verbascoside, isoverbascoside, and hispidulin were 248.40 ± 15.82, 393.10 ± 15.27, and 3.86 ± 0.87 µg/mL, respectively, indicating that the anti-proliferative effects of the C. chinense leaf extract mainly resulted from hispidulin and verbascoside. The selectivity index (SI) of C. chinense leaf extract against A549 lung cancer cells vs. normal keratinocytes were 2.4 and 2.8 after incubation for 24 and 48 h, respectively, suggesting the cytotoxic selectivity of the extract toward the cancer cell line. Additionally, the C. chinense leaf extract at 250 µg/mL induced late apoptotic cells up to 21.67% with enhancing reactive oxygen species (ROS) induction. Furthermore, the lung cancer cell colony formation was significantly inhibited after being treated with C. chinense leaf extract in a dose-dependent manner. The C. chinense leaf extract at 250 µg/mL has also shown to significantly inhibit cancer cell migration compared with the untreated group. The obtained results provide evidence of the anti-lung cancer potentials of the C. chinense leaf ethanolic extract.

Sustainable Downscaled Catalytic Colorimetric Determination of Manganese in Freshwater Using Smartphone-Based Monitoring Oxidation of 3,3',5,5'-Tetramethylbenzidine by Periodate.

A sustainable downscaled procedure using smartphone-based colorimetric determination of manganese (Mn(II)) was developed. This novel Mn(II) determination procedure is proposed using a simple, available microwell-plate platform and a smartphone as a detector. This approach is based on the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) by periodate using Mn(II) as a catalyst. The catalytic kinetics of Mn(II) under different conditions was investigated to determine the optimum condition where the different catalytic activities of various concentrations of Mn(II) evince. Under the optimum condition, the bluish-green product of oxidized TMB, proportioned to the concentration of Mn(II), was monitored using a smartphone camera, and the color signals were processed using ImageJ Software. The developed procedure showed great selectivity and sensitivity as linearity ranged from 1.8 × 10-6 to 4.6 × 10-5 M (0.1 to 2.5 µg/mL). The limits of detection and quantitation were 3.6 × 10-6 and 1.1 × 10-5 M (0.2 and 0.6 µg/mL), respectively. The determination of Mn(II) in freshwater samples was demonstrated to assess environmental water quality as an initial model to more easily promote water management according to the United Nations Sustainable Development Goals (UN-SDGs). The intensity of the red could be successfully applied to evaluate Mn(II) in canals and river water with no significant differences compared with the reference method of Inductively Coupled Plasma Optical Emission Spectrometry at a confidence level of 95%.

Anthocyanin Profile, Antioxidant, Anti-Inflammatory, and Antimicrobial against Foodborne Pathogens Activities of Purple Rice Cultivars in Northern Thailand.

Five glutinous purple rice cultivars and non-glutinous purple rice cultivated in different altitudes in the north of Thailand were collected. The samples were extracted using ethanol and determined for anthocyanins using HPLC. The total phenolic content (TPC), total flavonoid content (TFC), and the antioxidant, anti-inflammatory, and antimicrobial activities against foodborne pathogens were investigated. The highland glutinous cultivar named Khao' Gam Luem-Phua (KGLP) extract had significantly high levels of cyanidin 3-O-glucoside, peonidin 3-O-glucoside, delphinidin 3-O-glucoside, TPC, and TFC, as well as exerting a potent antioxidant activity through ABTS assay (524.26 ± 4.63 VCEAC, mg l-ascorbic-ascorbic/g extract), lipid peroxidation (IC50 = 19.70 ± 0.31 µg/mL), superoxide anions (IC50 = 11.20 ± 0.25 µg/mL), nitric oxide (IC50 = 17.12 ± 0.56 µg/mL), a suppression effect on nitric oxide (IC50 = 18.32 ± 0.82 µg/mL), and an inducible nitric oxide synthase production (IC50 = 23.43 ± 1.21 µg/mL) in combined lipopolysaccharide-interferon-γ-activated RAW 264.7 murine macrophage cells. Additionally, KGLP also exhibited antimicrobial activity against foodborne pathogens, Staphylococcus aureus, Escherichia coli, Salmonella Enteritidis, and Vibrio parahaemolyticus. These results indicate that Thai glutinous purple rice cultivated on the highland could be a potent natural source of antioxidants, anti-inflammatories, and antimicrobial agents for use as a natural active pharmaceutical ingredient in functional food and nutraceutical products.

Modulation of Intestinal Transport and Absorption of Topotecan, a BCRP Substrate, by Various Pharmaceutical Excipients and Their Inhibitory Mechanisms of BCRP Transporter.

Breast cancer resistance protein transporter (ABCG2/BCRP) is highly expressed on the intestinal epithelial membrane and has a significant impact on the oral absorption of topotecan. In this study, we examined 6 pharmaceutical excipients including BL-9EX, Brij97, Cremophor EL, Labrasol, Pluronic F68, and Tween 20 for their BCRP inhibitory effects. A bidirectional transport study using Caco-2 cells demonstrated that Tween 20 and Cremophor EL significantly increased the absorptive transport of topotecan, while simultaneously decreasing secretory transport. Interestingly, Labrasol selectively increased absorptive transport, whereas Pluronic F68 selectively decreased the secretory transport, of topotecan. Further investigation using an in situ closed loop experiment showed that 0.05% (w/v) Tween 20 and Cremophor EL significantly increased the intestinal absorption of topotecan in rats. An LDH assay demonstrated that 0.05% (w/v) Tween 20 and Cremophor EL did not cause significant damage to intestinal epithelial membranes. Furthermore, we examined the absorption-enhancing mechanisms of these excipients and found that Cremophor EL, Tween 20, and Labrasol increased the membrane fluidity of the inner lipid bilayers of the intestine. Therefore, this might be one of the most important mechanisms for inhibition of BCRP function by these excipients in the intestine.

Effects of Various Pharmaceutical Excipients on the Intestinal Transport and Absorption of Sulfasalazine, a Typical Substrate of Breast Cancer Resistance Protein Transporter.

Breast cancer resistance protein (BCRP) transporter is an efflux transporter that utilizes energy from adenosine triphosphate hydrolysis to push its substrates, regardless of the concentration gradient. Its presence on the apical membrane of the intestinal mucosa is a major obstacle for the intestinal absorption of its substrates. In this study, we examined the effects of various pharmaceutical excipients on the intestinal transport and absorption of sulfasalazine, a BCRP substrate. Four excipients, including 0.05% and 0.075% BL-9EX, 0.01% and 0.05% Brij 97, 0.075% Labrasol, and 0.05% and 0.1% Tween 20 decreased the secretory transport of sulfasalazine in an in vitro diffusion chamber. Further investigation in an in situ closed loop experiment in rats showed that 0.05% and 0.1% BL-9EX and 0.1% Brij 97 effectively enhanced the intestinal absorption of sulfasalazine while maintaining minimal toxicity to the intestinal mucosa. However, 0.1% Brij 97 also increased the intestinal absorption of 5(6)-carboxyfluorescein, a paracellular marker compound. These findings suggest that BL-9EX might effectively inhibit the BCRP-mediated efflux of sulfasalazine in vivo, indicating that BL-9EX could improve the intestinal absorption of sulfasalazine and other BCRP substrates.

Improvement of intestinal absorption of curcumin by cyclodextrins and the mechanisms underlying absorption enhancement.

Curcumin is known to possess a wide range of pharmacological activities for the treatment of chronic or inflammatory diseases, Alzheimer's disease, and various cancers. However, the therapeutic efficacy of curcumin is restricted by its poor bioavailability after oral administration. In this study, the effects of various cyclodextrins on the intestinal absorption of curcumin were evaluated in rat intestine by an in situ closed-loop method. Among the tested cyclodextrins, 50 mM α-cyclodextrin significantly enhanced the absorption of curcumin without inducing any intestinal toxicity. The analysis of cellular transport across Caco-2 cell monolayers showed that 50 mM α-cyclodextrin reduced the transepithelial electrical resistance value of cell monolayers and improved the permeability of 5(6)-carboxyfluorescein, a poorly absorbable drug, which is mainly transported via a paracellular pathway. Furthermore, the western blotting analysis showed that α-cyclodextrin decreased the expression of claudin-4, a tight junction-associated protein, in brush border membrane vesicles. Additionally, α-cyclodextrin increased the membrane fluidity of lipid bilayers in brush border membrane vesicles and may also have promoted the permeation of drug molecules via a transcellular pathway. These results suggested that α-cyclodextrin might enhance the intestinal absorption of curcumin via both paracellular and transcellular pathways.