ß-sitosterol isolated from the leaves of Trema orientalis (Cannabaceae) promotes viability and proliferation of BF-2 cells.

Trema orientalis is a pioneer species in the cannabis family (Cannabaceae) that is widely distributed in Thai community forests and forest edges. The mature leaves are predominantly used as an anti-parasite treatment and feed for local freshwater fish, inspiring investigation of their phytochemical composition and bioactivity. The purpose of this work was to investigate the bioactive compounds in T. orientalis leaf extract and their cytotoxicity in the BF-2 fish cell line (ATCC CCL-91). Flash column chromatography was used to produce 25 mL fractions with a mixture solvent system comprised of hexane, diethyl ether, methanol, and acetone. All fractions were profiled with HPLC-DAD (mobile phase methanol:aqueous buffer, 60:40 v/v) and UV detection (wavelengths 256 and 365 nm). After drying, a yellowish powder was isolated from lipophilic leaf extract with a yield of 280 µg/g dry weight. Structure elucidation by nuclear magnetic resonance (NMR) indicated it to consist of pure ß-sitosterol. The lipophilic extract and pure compound were evaluated for cytotoxicity using BF-2 cells. MTT assays showed both leaf extract and pure compound at 1 µg/mL to increase cell viability after 24 h treatment. The respective half maximal inhibitory concentration (IC50) values of leaf extract and ß-sitosterol were 7,027.13 and 86.42 µg/ml, indicating a lack of toxicity in the BF-2 cell line. Hence, T. orientalis can serve as a source of non-toxic natural lipophilic compounds that can be useful as bioactive ingredients in supplement feed development.

Lasianthus ranongensis (Rubiaceae), a new species from Andaman tropical rainforest Thailand.

We describe Lasianthus ranongensis Sinbumr. & Napiroon as a new species in the genus Lasianthus. The new taxon is intensively discussed through taxonomic affinities and information on its habitat, distribution and conservation status is provided. Moreover, line drawings and stereo microscope images of important fertile organs are demonstrated. The new species is morphologically similar to L. stipularis but differs in its having flattened branches (vs. terete), leaf blade elliptic-oblong shape of 15.0-20.0 × 4.0-6.0 cm (vs. oblanceolate-oblong 12.0-16.0 × 3.0-5.5 cm), 9-12 pairs of veins (vs. 9-10 pairs), stipule 5-7 mm long, half covering cymes (vs. 10-12.5 mm long, entirely covering cymes), four or five bracts narrowly lanceolate, 2.5-3.0 mm long (vs. bracts numerous, broadly triangular, 10-15 mm), flowers with cupular calyx (vs. with campanulate calyx), corolla villous on internal surface, and six or seven lobed (vs. pubescent on internal surface and four lobed) and drupes with five pyrenes (vs. with four pyrenes). It is also similar to L. pseudo-stipularis, but from which it is obviously distinguished by its stipule half covers cymes, secondary veins have 9-12 pairs of secondary veins, cupular calyx shape, six or seven lobed corolla, and drupe with five pyrenes, whereas L. pseudo-stipularis has stipule entirely covering cymes, 7-8 pairs of secondary veins, obconic calyx, four lobed corolla, and drupe with four pyrenes.

Cannabinoids from inflorescences fractions of Trema orientalis (L.) Blume (Cannabaceae) against human pathogenic bacteria.

BACKGROUND: Cannabinoids; tetrahydrocannabinol (THC), cannabidiol (CBD) and cannabinol (CBN), might show antibacterial activity. Trema orientalis is a species in the Cannabaceae that is closely related to Cannabis through plastome phylogenetic evidence. This species is widely distributed throughout tropical Asia and is used as traditional medicine, particularly for the treatment of infectious diseases. However, no studies on the antibacterial activity of cannabinoid-containing inflorescences extracts are available. Thus, the aim of this study was to determine cannabinoid content and antibacterial activity of inflorescences fractions from T. orientalis native to Thailand. METHODS: We hypothesized that inflorescences from T. orientalis might display cannabinoids similar to Cannabis because of their close taxonomic relationship. We extracted the mature inflorescences and infructescence of T. orientalis in three disparate populations from different Thailand floristic regions. Extractions were subsequently partitioned into hydrophilic and lipophilic fractions using distilled water and chloroform. The lipophilic extracts were further fractionated by the column chromatography with gradient elution and analyzed by gas chromatography-mass spectrometry (GC-MS). Characterized cannabinoids were used in bioassays with multidrug-resistance bacteria. RESULTS: Lipophilic extracts and fractions of inflorescences from all Thailand floristic regions consistently displayed cannabinoids (THC, CBD and CBN) in various quantities. These extracts exhibited inhibitory activity for Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii strains with minimum inhibitory concentration values varying from 31.25 to 125 µg/mL. CONCLUSION: Our study is the first to report cannabinoid detection in extracts from inflorescences of T. orientalis, a species in the Cannabaceae. These extracts and their fractions containing cannabinoids showed pronounced antibacterial activity. The use of analytic methods also demonstrated reproducible cannabinoid extraction.

ANTIBACTERIAL PROPERTY TESTING OF TWO SPECIES OF TROPICAL PLANT LASIANTHUS (RUBIACEAE).

Two tropical plant species, Lasianthus pilosus and Lasianthus stipularis are used in traditional medicine but there have been no published studies of the extracts of these plants against bacteria. In this study, we aimed to determine the antimicrobial activities of the above two plants against two gram-positive and seven gram-negative bacterial strains to determine the potential of these two plant species for possible antimicrobial drug development. The antibacterial activities of the lipophilic extracts of these plants were evaluated by disk diffusion and broth microdilution methods. The zone diameters and minimum inhibitory concentrations (MIC) for these plant extracts exhibited their highest antibacterial activity against Pseudomonas aeruginosa, followed by Staphylococcus aureus and Acinetobacter baumannii, respectively. The MIC of these extracts against P. aeruginosa (ATCC 37166 and ATCC 27853) varied from 50 to 200 µg/ml. Thin layer chromatography and detection using different specific reagents revealed the presence of terpenoids, phenolic compounds and iridoid. Cell lysis due to the effect of the lipophilic extracts of these tested plants was demonstrated using scanning electron microscopy. In conclusion, the bioactive compounds of these plants should be studied further to develop potential antimicrobial agents.