Bioactive Sesquiterpene Lactones Isolated from the Whole Plants of Vernonia cinerea.

Twelve sesquiterpene lactones were isolated from the whole plants of Vernonia cinerea. These included eight new compounds, vercinolides A-H (1-8), along with four known substances (9-12). The structures of the new compounds were determined by 1D and 2D NMR experiments and mass spectrometric methods. The absolute configurations of compounds 1-8 were determined by Mosher experiments and ECD data. Compounds 1-8 are the first examples of a new class of sesquiterpene lactones possessing a rare 4α,10α-ether ring and a 2,14-ether ring. Compounds 1-4, 6, 8, 10, and 12 were evaluated for their cytotoxic and anti-inflammatory activities. Compounds 10 and 12 exhibited inhibitory effects against nitric oxide production in lipopolysaccharide-activated RAW 264.7 mouse macrophage cells with IC50 values of 21 and 23 µM, respectively. Both compounds were inactive for HeLa cells (IC50 > 10 µM).

Optimal condition of cannabis maceration to obtain the high cannabidiol and Δ9-tetrahydrocannabinol content.

The aim of this work was to optimize a maceration condition of cannabis (Cannabis sativa L.). A circumscribed central composite experimental design was applied in this work. Temperature and time were varied from 40-80 °C and 30-90 min, respectively. The three responses (i.e., extraction yield, cannabidiol content, and Δ9- tetrahydrocannabinol content) were predicted by computer software. The yield was high when cannabis was macerated using ethanol at high temperature and long duration time. While cannabidiol and Δ9- tetrahydrocannabinol content was high when macerating at a low heating temperature and short duration time. The optimal condition provided the simultaneous high of cannabidiol and Δ9- tetrahydrocannabinol content was 40 °C for 30 min. The prediction was accurate due to low percent error. This optimal condition could be used as a guide for maceration of cannabis to obtain the extract containing a high content of cannabidiol and Δ9- tetrahydrocannabinol.

Fabrication of Orally Fast-Disintegrating Wafer Tablets Containing Cannabis Extract Using Freeze-Drying Method.

Introduction: The development of a novel dosage form for cannabis extract is necessary to improve drug delivery and also enhance patient convenience. Methods: Orally fast-disintegrating wafer tablets containing cannabis extract, which were prepared using the freeze drying technique, were developed in this work. The formulation consisted of several key components: cannabis extract as the active compound, Tween® 80 as a surfactant and solubilizer, gelatin and mannitol as structural components, sucralose as a sweetening agent, and sodium methylparaben and sodium propylparaben as preservatives. Results: The optimized formulation consists of the following ingredients: 5% cannabis extract, 1.25% Tween® 80, 5% gelatin, 88.34% mannitol, 0.2% sucralose, 0.19% sodium methylparaben, and 0.02% sodium propylparaben. The resulting wafer tablets exhibited the following characteristics: a porous structure, an average weight of approximately 200 mg, minimal weight variation (less than 1.4%), slightly acidic pH (pH 5.12), disintegration within 10 s, low moisture content (less than 3%), a Δ9-tetrahydrocannabinol content of approximately 2.8 mg, and a cannabidiol content of approximately 0.9 mg. Additionally, the wafer tablets rapidly dissolved in simulated saliva fluid containing sodium lauryl sulfate. Conclusion: This work succeeded in the fabrication of orally fast-disintegrating wafer tablets containing cannabis extract with desired properties.

Community Culture Survey - Revised: Measuring neighborhood culture and exploring geographic, socioeconomic, and cultural determinants of health in samples across the United States and in Thailand.

Objectives: Research on links between social, geographic, and cultural determinants of health has been thwarted by inadequate measures of culture. The purpose of this study was to improve the measurement of community culture, defined as shared patterns of attitudes and behaviors among people within a neighborhood that distinguish it from others, and to examine dimensions of culture, independent of socioeconomic and demographic factors, and their relationships with health. Study design: A survey research design with correlational analyses was used. Methods: A survey packet including the Community Culture Survey - Revised (CCS-R), demographic, health, and other individual-level measures was administered through convenience sampling across the United States (US) and to a sample in Thailand from 2016 to 2018. US county-level variables were obtained from zip codes. Results: 1930 participants from 49 US states (n = 1592) and Thailand (n = 338) completed all CCS-R items, from which 12 subscales were derived: Social Support & Connectedness, Responsibility for Self & Others, Family Ties & Duties, Social Distress, Urban Diversity, Discontinuity, Church-Engaged, External Resource-Seeking, Locally Owned Business-Active, Power Deference, Next Generation Focus, and Self-Reliance. Neighborhood culture subscale scores varied more by geography than by participant's demographics. All subscales predicted one or more health indicator, and some of these relationships were significant after adjusting for participant age and county-level socioeconomic variables. Most of the significant differences on subscales by race/ethnicity were no longer significant after adjusting for participant's age and county-level socioeconomic variables. Most rural/urban and regional differences in culture within the US persisted after these adjustments. Based on correlational analyses, Social Support & Connectedness and Responsibility for Self & Others were the best predictors of participants' overall health and quality of life, and Responsibility for Self & Others was the best predictor (inversely) of the CDC's measures of social vulnerability. Conclusions: Neighborhood culture is measurable, multi-dimensional, distinct from race/ethnicity, and related to health even after controlling for age and socioeconomic factors. The CCS-R is useful for advancing research and practice addressing the complex interactions between individuals, their neighborhood communities, and health outcomes.

In vitro antioxidant, anti-inflammatory, and anticancer activities of mixture Thai medicinal plants.

BACKGROUND: The phytochemical study of medicinal plants is rapidly gaining popularity with many pharmacologic effects. This study aims to determine the antioxidant capacity as well as anticancer and antimigration activities of Clear belongs Plus extract (CBL-P) which consisted of five medicinal plants namely, Alpinia galanga, Piper nigrum, Citrus aurantifolia, Tiliacora triandra, and Cannabis sativa on human colon cancer cells SW620 and HCT116 cell lines, and human non-small cell lung cancer cells A549 and NCI-H460 cell lines. METHODS: In this study the dried-plant powder was extracted using 90% ethanol. Additionally, CBL-P was studied antioxidative activity via DPPH and ABTS assays and anti-inflammatory activities using nitric oxide assay using Griess reaction. Antiproliferation and antimigration of CBL-P were investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and scratch assay. RESULTS: The results showed that CBL-P had potent antiproliferative activity with IC50 values in a concentration- and time-dependent manners for all four cell lines. CBL-P also possessed potent antimigration activity against all studied cancer cells. CBL-P demonstrated antimigration activity on four different types of cancer cells (A549, NCI-H460, HCT116, and SW620) after 48 h of incubation, with the greatest effect seen at the highest concentration tested (15 µg/mL) in A549 cells (10.23% of wound closure) and NCI-H460 cells (9.16% of wound closure). CBL-P was also effective in reducing migration in HCT116 and SW620 cells, with a range of closure area from 10-50%. In addition, CBL-P had antioxidant activity with IC50 values of 8.549 ± 0.241 mg/mL and 2.673 ± 0.437 mg/mL for DPPH and ABTS assays, respectively. CBL-P also showed anti-inflammatory activity with the best inhibitory activity on NO production at a concentration of 40 µg/mL. CONCLUSION: In conclusion, the mixture extract possessed antioxidant and anti-inflammatory activity. Furthermore, the mixture plant extract significantly exhibited antiproliferative and antimigration activities on SW620, HCT116, A549, and NCI-H460 cells (P ≤ 0.05). Taken together, our results suggest that medicinal plants may have synergistic effects that could potentially enhance the effectiveness of cancer treatment when used as adjuvants. These findings provide a solid scientific foundation for future efforts to explore the mechanism of action.

Anti-inflammatory sesquiterpene lactones from the flower of Vernonia cinerea.

Bioassay-guided fractionation of the hexane extract from the flowers of Vernonia cinerea (Asteraceae) led to the isolation of a new sesquiterpene lactone, 8α-hydroxyhirsutinolide (2), and a new naturally occurring derivative, 8α-hydroxyl-1-O-methylhirsutinolide (3), along with seven known compounds (1 and 4-9). The structures of the new compounds were determined by 1D and 2D NMR experiments and by comparison with the structure of compound 1, whose relative stereochemistry was determined by X-ray analysis. The isolated compounds were evaluated for their cancer chemopreventive potential based on their ability to inhibit nitric oxide (NO) production and tumor necrosis factor alpha (TNF-α)-induced NF-κB activity. Compounds 1, 2, 4, 5, and 9 inhibited TNF-α-induced NF-κB activity with IC(50) values of 3.1, 1.9, 0.6, 5.2, and 1.6 µM, respectively; compounds 4 and 6-9 exhibited significant NO inhibitory activity with IC(50) values of 2.0, 1.5, 1.2, 2.7, and 2.4 µM, respectively.

Antitumor Effects of Delta (9)-Tetrahydrocannabinol and Cannabinol on Cholangiocarcinoma Cells and Xenograft Mouse Models.

Cholangiocarcinoma (CCA) is a very aggressive tumor. The development of a new therapeutic drug for CCA is required. This study aims to evaluate the antitumor effect of ∆9-tetrahydrocannabinol (THC), the major psychoactive component of marijuana (Cannabis sativa), and cannabinol (CBN), a minor, low-psychoactive cannabinoid, on CCA cells and xenograft mice. THC and CBN were isolated, and their identities were confirmed by comparing 1H- and 13C-NMR spectra and mass spectra with a database. Cell proliferation, cell migration, and cell apoptosis assays were performed in HuCCT1 human CCA cells treated with THC or CBN. The phosphorylation of signaling molecules in HuCCT1 cells was detected. To determine the effects of THC and CBN in an animal model, HuCCT1 cells were inoculated subcutaneously into nude mice. After the tumors reached an appropriate size, the mice were treated with THC or CBN for 21 days. Tumor volumes were monitored and calculated. The 1H- and 13C-NMR data of THC and CBN were almost identical to those reported in the literature. THC and CBN significantly inhibited cell proliferation and migration and induced apoptosis in HuCCT1 cells. The phosphorylation of AKT, GSK-3α/ß, and ERK1/2 decreased in HuCCT1 cells treated with THC or CBN. CCA xenograft mice treated with THC showed significantly slower tumor progression and smaller tumor volumes than control mice. THC and CBN induced apoptosis in CCA by inhibiting the AKT and MAPK pathways. These findings provide a strong rationale for THC and CBN as therapeutic options for CCA.

[Using liquid chromatography-mass spectrometry based metabolomics to discriminate between cold pressed rice bran oils produced from two different cultivars of Oryza sativa L. ssp. indica in Thailand].

A newly developed liquid chromatography-mass spectrometry (LC-MS) method for the analysis of cold pressed rice bran oil (RBO) was established and used to discriminate between RBOs produced from two different cultivars of major Thai fragrant rice species. The cold pressed RBO was prepared using the screw compression method. The LC-MS data were preprocessed with MZmine 2.10 program before evaluating with principal component analysis using SIMCA 13 software. The LC-MS method was able to detect and quantify several kinds of valuable constituents such as fatty acids, vitamin E, and γ-oryzanol. The chromatographic condition was feasible; short time for analysis and simple method were achieved. From score plot and loading plot of principle component analysis (PCA) , two rice cultivar samples were clearly separated, and it was revealed that Khao-Hom-Pathum was more suitable than Khao-Hom-Mali for cold pressed RBO production since it contained high total γ-oryzanol and less saturated free fatty acids. As with the fixed price of all the rice brans, this information can be used in order to, if possible, preserve the price of rice brans from different cultivars.

An optimization approach of dynamic maceration of Centella asiatica to obtain the highest content of four centelloids by response surface methodology

ABSTRACT Centella asiatica (L.) Urb., Apiaceae, is commonly used as food, food supplement, and medicine. Development of the extraction process to obtain the high extent of the active compound is necessary. So, the response surface methodology was used in this work to optimize the dynamic maceration of C. asiatica to obtain the highest content of the four centelloids including asiatic acid, madecassic acid, asiaticoside, and madecassoside. Two factors: extraction temperature and extraction time, were studied. The content of four centelloids was observed. After the extraction of C. asiatica using ethanol, the content of four centelloids was analyzed using validated high performance liquid chromatography. The optimization result showed that madecassoside and asiaticoside had a similar pattern of the contour plots and response surfaces. These two centelloids were highly extracted at a high extraction time and high extraction temperature. The other two centelloids had the same pattern, they had a high content at a high temperature and time as well as at a low temperature and time. The simultaneous highest content of four centelloids was achieved when extracted at 60 °C for 120 min. The optimal condition could be used as standard condition for extraction of C. asiatica to provide the highest content of four centelloids.

Bioactive sesquiterpene lactones and other compounds isolated from Vernonia cinerea.

Four new sesquiterpene lactones, 8α-(2'Z-tigloyloxy)-hirsutinolide (1), 8α-(2'Z-tigloyloxy)-hirsutinolide-13-O-acetate (2), 8α-(4-hydroxytigloyloxy)-hirsutinolide (3), and 8α-hydroxy-13-O-tigloyl-hirsutinolide (4), along with seven known derivatives (5-11), three norisoprenoids (12-14), a flavonoid (15), and a linoleic acid derivative (16), were isolated from the chloroform partition of a methanol extract from the combined leaves and stems of Vernonia cinerea. Their structures were established by 1D and 2D NMR, UV, and MS analyses. Compounds 1-16 were evaluated for their inhibitory effects against the viability of U251MG glioblastoma and MDA-MB-231 breast cancer cells that harbour aberrantly-active STAT3, compared to normal NIH3T3 mouse fibroblasts that show no evidence of activated STAT3. Among the isolates, compounds 2 and 7 inhibited the aberrant STAT3 activity in glioblastoma or breast cancer cells. Further, compounds 7 and 8 inhibited viability of all three cell lines, compounds 2, 4, and 9 predominantly inhibited the viability of the U251MG glioblastoma cell line.

Validated high performance liquid chromatography for simultaneous determination of stability of madecassoside and asiaticoside in film forming polymeric dispersions

ABSTRACT The objective of the work was to validate the high performance liquid chromatography for simultaneous determination of stability of madecassoside and asiaticoside in Centella asiatica (L.) Urb., Apiaceae, extract-loaded film forming polymeric dispersions. High performance liquid chromatography method was validated in five topics: linearity and range, limit of detection and limit of quantitation, specificity, precision, and accuracy. Results showed the method had a good linearity (R2 > 0.9990) in the range of 5-150 µg/ml and specific. The limit of detection and limit of quantitation of madecassoside were 81 and 245 ng/ml and asiaticoside were 21 and 64 ng/ml, respectively. The percent relative standard deviation of intraday and interday precision were less than 1 and 3%, respectively. The accuracy presented as percent recovery was 101.54-103.29% for madecassoside and 100.39-102.58% for asiaticoside. This validated high performance liquid chromatography method was used to determine the stability of the formulation containing Centella asiatica extract. Centella asiatica extract-loaded film forming polymeric dispersions used Eudragit® RS 30D and Eudragit® RL 30D as film former, glycerin as plasticizer, and absolute ethanol as solvent and penetration enhancer. Three formulations with different ratio of Eudragit® RS 30D and Eudragit® RL 30D were prepared and stored for 90 days at 4 ºC, 25 ºC, and 40 ºC. Stability results showed that almost all of the formulations were unstable at 25 ºC and 40 ºC. Except, two of three formulations were stable at 4 ºC. However, the formulation was further developed to improve the stability of madecassoside and asiaticoside in the formulation.

Lipid-lowering and antioxidative activities of aqueous extracts of Ocimum sanctum L. leaves in rats fed with a high-cholesterol diet.

The present study was conducted to investigate the lipid-lowering and antioxidative activities of Ocimum sanctum L. (OS) leaf extracts in liver and heart of rats fed with high-cholesterol (HC) diet for seven weeks. The results shows that OS suppressed the high levels of serum lipid profile and hepatic lipid content without significant effects on fecal lipid excretion. Fecal bile acids excretion was increased in HC rats treated with OS. The high serum levels of TBARS as well as AST, ALT, AP, LDH, CK-MB significantly decreased in HC rats treated with OS. OS suppressed the high level of TABARS and raised the low activities of GPx and CAT without any impact on SOD in the liver. As for the cardiac tissues, OS lowered the high level of TABARS, and raised the activities of GPx, CAT, and SOD. Histopathological results show that OS preserved the liver and myocardial tissues. It can be concluded that OS leaf extracts decreased hepatic and serum lipid profile, and provided the liver and cardiac tissues with protection from hypercholesterolemia. The lipid-lowering effect is probably due to the rise of bile acids synthesis using cholesterol as precursor, and antioxidative activity to protect liver from hypercholesterolemia.

Antioxidant Activity and Lipid-Lowering Effect of Essential Oils Extracted from Ocimum sanctum L. Leaves in Rats Fed with a High Cholesterol Diet.

It has been reported that Ocimum sanctum L. (OS) leaves decrease serum lipid profile in normal and diabetic animals. No experimental evidences support the anti-hyperlipidemic and antioxidative actions against hypercholesterolemia. Moreover the identity of the specific chemical ingredients in OS leaves responsible for these pharmacological effects are unknown. Since OS leaves are rich in essential oil (EO). Therefore the present study was conducted to investigate the anti-hyperlipidemic and antioxidative activities of EO extracted from OS leaves in rats fed with high cholesterol (HC) diet. EO was extracted by the hydrodistillation method and the chemical constituents were then identified by Gas Chromatography-Mass Spectrometry. The experiment was performed in Male Wistar rats fed with 2.5 g%(w/w) of cholesterol diet for seven weeks. During the last 3 weeks, rats were daily fed with EO. The results showed that phenyl propanoid compounds including eugenol and methyl eugenol were the major constituents of EO. EO suppressed the high serum lipid profile and atherogenic index as well as serum lactate dehydrogenase and creatine kinase MB subunit without significant effect on high serum levels of aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase in rats fed with HC diet. In addition, EO was found to decrease the high levels of thiobarbituric acid reactive substances (TBARS), glutathione peroxidase (GPx) and superoxide dismutase (SOD) without impacting catalase (CAT) in the cardiac tissue while in the liver, it decreased high level of TBARS without significantly effecting GPx, SOD and CAT. Histopathological results confirmed that EO preserved the myocardial tissue. It can be concluded that EO extracted from OS leaves has lipid-lowering and antioxidative effects that protect the heart against hypercholesterolemia. Eugenol that is contained in EO likely contribute to these pharmacological effects.