Noppamas Soonthornchareonnon

Antitumour effects of phyllanthus emblica L.: Induction of cancer cell apoptosis and Inhibition of in vivo tumour promotion and in vitro invasion of human cancer cells

Phyllanthus emblica Linn. (PE) is a medicinal fruit used in many Asian traditional medicine systems for the treatment of various diseases including cancer. The present study tested the potential anticancer effects of aqueous extract of PE in four ways: (1) against cancer cell lines, (2) in vitro apoptosis, (3) mouse skin tumourigenesis and (4) in vitro invasiveness. The PE extract at 50–100 µg/mL significantly inhibited cell growth of six human cancer cell lines, A549 (lung), HepG2 (liver), HeLa (cervical), MDA‐MB‐231 (breast), SK‐OV3 (ovarian) and SW620 (colorectal). However, the extract was not toxic against MRC5 (normal lung fibroblast). Apoptosis in HeLa cells was also observed as PE extract caused DNA fragmentation and increased activity of caspase‐3/7 and caspase‐8, but not caspase‐9, and up‐regulation of the Fas protein indicating a death receptor‐mediated mechanism of apoptosis. Treatment of PE extract on mouse skin resulted in over 50% reduction of tumour numbers and volumes in animals treated with DMBA/TPA. Lastly, 25 and 50 µg/mL of PE extract inhibited invasiveness of MDA‐MB‐231 cells in the in vitro Matrigel invasion assay. These results suggest P. emblica exhibits anticancer activity against selected cancer cells, and warrants further study as a possible chemopreventive and antiinvasive agent. Copyright

Rapid micropropagation of Curcuma longa using bud explants pre-cultured in thidiazuron-supplemented liquid medium

Multiple shoots of Curcuma longa were induced by culture of bud explants for 1 week in Murashige and Skoog (MS) liquid medium supplemented with 72.64 μM thidiazuron (TDZ) prior to culture on MS gelled medium without growth regulator for 8 weeks. The regeneration rate was up to 11.4 ± 1.7 shoots/explant. Rooting was spontaneous and the regenerated plants were successfully transferred to soil. This protocol can be an alternative for rapid micropropagation of C. longa used for phytomedicine raw material production.

Partial depolymerization of pectin by a photochemical reaction

Complex heterogeneous polysaccharides that comprise pectin were partially depolymerized by a photochemical reaction using ultraviolet light in the presence of titanium dioxide catalyst. In a period of 6h at pH 7, this UV/TiO(2) process decreased the average molecular weight of pectin from 400kDa to 200kDa. The characterization of the partially depolymerized pectin, which was fractionated by size-exclusion chromatography, was performed by (1)H NMR spectroscopy, and the spectra obtained showed that the resulting oligosaccharides and polysaccharides maintained the intact core structure of pectin. The monosaccharide content and depolymerization profile were determined by high-performance anion-exchange chromatography coupled with pulsed amperometric detection. This controlled photochemical depolymerization technique might be useful for preparation of pectin oligosaccharides as an ingredient in food and pharmaceutical products.

Aurocitrin and Related Polyketide Metabolites from the Wood-Decay Fungus Hypocrea sp. BCC 14122

The known gentisaldehyde antibiotic aurocitrin (1), its cis-olefinic isomer (2), a salicylaldehyde analogue (3), two new benzofuran derivatives (4 and 5), and a new dihydroisocoumarin (6) were isolated from the wood-decay fungus Hypocrea sp. BCC 14122. The structures were elucidated primarily by NMR and mass spectroscopic analyses.

Estrogenic activity of the dichloromethane extract from Pueraria mirifica

Pueraria mirifica and its extracts are widely used as the ingredient(s) in many rejuvenating products. Up to now, the extract of P. mirifica roots that has been used in most studies, is the alcoholic extract. In the present study, we investigated the estrogenic activity using uterotropic and MCF-7 cell proliferation models of the dichloromethane extract as well as the water extract which was obtained from partitioning the ethanolic extract. The results indicated that among the three extracts, i.e. the ethanolic extract (PM1), the water extract (PM2) and dichloromethane extract (PM3), PM3 exhibited the most potent estrogenic activity in both models, followed by PM1. The extracts produced uterotropic activity associated with the increase of water content while uterotropic activity of 17beta-estradiol was related to the increase of muscle mass. The two isoflavonoids, genistein and daidzein, were not the major active phytoestrogens involving the estrogenic activity of these extracts.

Alkaline phosphatase activity-guided isolation of active compounds and new dammarane-type triterpenes from Cissus quadrangularis hexane extract

ETHNOPHARMACOLOGICAL RELEVANCE The stem of Cissus quadrangularis L. (CQ) is used in traditional medicine to treat bone fractures and swelling. Anti-osteoporotic activity of CQ hexane extract has been reported, but the active compounds in this extract remain unknown. Thus, we aimed to identify the active compounds in CQ hexane extract using bioassay-guided isolation. MATERIALS AND METHODS The CQ hexane extract was fractionated sequentially with benzene, dichloromethane, ethyl acetate, and methanol. The examination of CQ extract and its fractions was guided by bioassays for alkaline phosphatase (ALP) activity during the differentiation of MC3T3-E1 osteoblastic cells. The cells were treated with or without the CQ extract and its fractions for a period of time, and then the stimulatory effect of the alkaline phosphatase enzyme, a bone differentiation marker, was investigated. The compounds obtained were structurally elucidated using spectroscopic techniques and re-evaluated for activity during bone differentiation. RESULTS A total of 29 compounds were isolated, viz., triterpenes, fatty acid methyl esters, glycerolipids, steroids, phytols, and cerebrosides. Four new dammarane-type triterpenes were isolated for the first time from nature, and this report is the first to identify this group of compounds from the Vitaceae family. Seven compounds, viz., glycerolipids and squalene, stimulated ALP activity at a dose of 10μg/mL. Moreover, the synergistic effect of these compounds on bone formation was demonstrated. CONCLUSION This report describes, for the first time, the isolation of active compounds from CQ hexane extract; these active compounds will be useful for the quality control of extracts from this plant used to treat osteoporosis.

A preliminary study of the acaricidal activity of clove oil, Eugenia caryophyllus.

BACKGROUND The search for more eco-friendly acaricides has prompted testing of medicinal plants from botanical sources. OBJECTIVE To evaluate the eradication of house dust mites (HDM), Dermatophagoides pteronyssinus, by direct contact using the essential clove oil (Eugenia caryophyllus). METHODS A pilot study was initiated to determine the killing power of clove oil. Synthetic fibers were immersed in 2% clove oil for 30 min, dried in a hot air oven at 60°C for 2 hrs after which 0.5 gm of HDMs were exposed to these coated fibers placed in the Siriraj Chamber (SC). Two additional long-term methods were employed. Ten mites were placed in the SC and 10 μl of clove oil was pipetted or sprayed onto them. These latter two procedures were each carried out for 3 consecutive days at 0, 1, 3 and 6 months. The solutions antimicrobial and antifungal properties were evaluated by exposing common bacteria and fungi to sterile filter disks impregnated with the mixture, and after overnight incubation, the disc diffusion method on nutrient agar was used. Ethyl alcohol served as the placebo. 99% and 81%, respectively, while the placebo mortality was <5%. The zone of inhibition indicated significant clearance for all the bacteria and fungi indicating greater biocidal activity when compared to the controls. RESULTS SEMs revealed dead mites on the fibers. The effectiveness of pipetting and spraying was 99% and 81%, respectively, while the placebo mortality was <5%. The zone of inhibition indicated significant clearance for all the bacteria and fungi indicating greater biocidal activity when compared to the controls. CONCLUSIONS Clove oil is a promising agent for killing dust mites with a potential use in dust-mite laden mattresses. Spraying diminishes in efficiency after 3 months.

Characterisation and immuno-stimulating activity of polysaccharides from Thai medicinal plants

Water-soluble polysaccharides were isolated from the tubers of Butea superba Roxb. and Pueraria candollei Wall. Ex Benth. var. mirifica (Shaw et Suvat.) C. Niyomdham, the leaves of Centella asiatica (L.) Urb, Ocimum basilicum L., Psidium guajava and Andrographis paniculata (Burn. f.) Nees, the stems of Cymbopogon citratus (Stapf ExG), and the fruits of Psidium guajava and Scaphium scaphigerum. The immunological impacts of the polysaccharides on T-lymphocyte proliferation in vitro was investigated by flow cytometric (immunofluorescence) analysis using staphylococcal enterotoxin B (SEB) as a positive control. It was found that the polysaccharides enhanced T-lymphocyte proliferation, ranging from 4.5 to 27.0% at a concentration of 100 µg mL−1, while the activity of SEB was 13.3%. The medicinal plants showing the highest immuno-stimulating activity were the tubers of Butea superba Roxb. The water-extracted tubers contained 60.0% (w/w) carbohydrates with 6.6% (w/w) uronic acid. The major constituent monosaccharides of the tubers were 28.2 mol% galactose, 10.5 mol% arabinose and 36.4 mol% glucose.

Efficacy and safety of “Yahom” as a traditional Thai herbal therapy: a systematic review

ETHNOPHARMACOLOGICAL RELEVANCE Yahom is a traditional Thai medicine used to treat syncope and abdominal discomfort. AIM OF THE STUDY This study aimed to systematically review all available evidence which purports to support these claims. MATERIAL AND METHODS The systematic review accorded with the Cochrane Collaboration framework and PRISMA reporting. Databases including MEDLINE, Excerpta Medica Database (EMBASE), Cochrane library database, and Google Scholar were searched by keywords, Yahom and Ya-hom. Pharmacological and toxicity data from non-animal and animal studies were included. RESULTS Twenty-four articles: 2 on in vitro cell lines or bacteria, 3 in vitro cell-free, 5 in vitro animal, 13 in vivo and 1 human mainly reported (A) Cardiovascular effects (i) transient hypotension (0.2-0.8g/kg, intravenous injection (i.v.)), increased cerebral blood flow (2g/kg, single oral) and vascular dilatation/relaxation (ii) elevated blood pressure (BP) (0.2-0.8g/kg, i.v. or 2-4g/kg oral) and vasocontraction. Single Yahom doses (3g) given to healthy volunteers had no effect on cutaneous blood flow, ECG or systolic BP although marginally increased diastolic BP was claimed. (B) Yahom (2-4g/kg) completely inhibited gastric acid secretion evoked by gastric secretagogues

Antiplasmodial activities of a Thai traditional antipyretic formulation, Bencha-Loga-Wichian: A comparative study between the roots and their substitutes, the stems

ETHNOPHARMACOLOGICAL RELEVANCE Bencha-Loga-Wichian (BLW) is a polyherbal antipyretic formulation that is comprised of Capparis micracantha, Clerodendrum indicum, Ficus racemosa, Harrisonia perforata, and Tiliacora triandra. A traditional medical textbook has documented the use of this formulation for the treatment of many types of fever, including malaria-like fever. Traditionally, BLW is composed of the root parts of those plants. However, in current practice, the stem parts are frequently substituted. Thus, this study aimed to evaluate the antiplasmodial activities of BLW and compare the efficacy between the stem and root parts. MATERIALS AND METHODS BLW formulations produced from either the stem or root parts of the various constituent plants as well as the stems or roots of the individual plants were separately extracted and tested against the chloroquine-sensitive (Pf3D7) and -resistant (PfW2) strains Plasmodium falciparum using flow cytometry. The cytotoxicity against peripheral blood mononuclear cells was evaluated using the WST-8 assay to determine the selectivity index (SI). The active compounds of BLW were isolated using antiplasmodial-guided isolation and quantified using Ultra-Performance Liquid Chromatography (UPLC). RESULTS The stem and root parts of BLW and the individual plants exhibited antiplasmodial activities at the same levels with good SI values in the range of 3.55-19.74. The extracts of BLW exhibited promising antiplasmodial activity against both Pf3D7 (IC50<5µg/mL) and PfW2 (IC50=6-10µg/mL). Among the five component plants, T. triandra was the most active and exhibited an IC50<5µg/mL against both strains of parasites with SI values >10. We isolated tiliacorinine and yanangcorinine as the major active compounds (IC50<2µg/mL). However, these two compounds demonstrated cytotoxic effects (SI<1). The UPLC analysis identified these compounds in both the stem and root parts of BLW in the range of 0.57-7.66%, which correlated with the antiplasmodial activity. The concentrations of these compounds in BLW, at comparable efficacy, were much less than those at the IC50s for the single compounds alone. It suggested that synergistic interactions increased the antiplasmodial effects as well as alleviated the toxicity of the active compounds in BLW. CONCLUSION This study described a promising antiplasmodial activity of BLW that had good selectivity and a toxicity-alleviating effect. The results provided scientific support for the use of this formulation for the treatment of malaria. In addition, the stem and root parts of the plants in BLW exhibited equivalent activities, which indicates the potential for the substitution of the stem parts in the formulation. Thus, we recommend additional study of the stem parts of these plants for further development on the basis of the availability and sustainability.