Wildfires are common in seasonally dry parts of the world with a Mediterranean climate. Prescribed burning is used to reduce fuel load and fire risk, but often without reliable information on its effects. This study investigated the effects of prescribed burns in different seasons on Pterostylis revoluta, an autumn-flowering Australian terrestrial orchid, and its orchid mycorrhizal fungi (OMFs) to find the least damaging season for a prescribed burn. Burns were conducted mid-season in spring and summer 2011 and autumn and winter 2012. Orchids were enumerated and measured during their flowering season in autumn 2011–2014 and mycorrhizal fungi were isolated before and after the burns in autumn 2011, 2012 and 2014. Micro-organisms isolated were characterized. DNA was extracted from the OMFs, and the internal transcribed spacer region was amplified by PCR. Amplicons were clustered by restriction fragment length polymorphism (RFLP), and representative amplicons were sequenced. OMF were tested for sensitivity to smoke water. The number of plants increased up to 4-fold and 90 % of plants became vegetative during this study. Isolation of mycorrhizal fungi increased and isolation of bacteria decreased. Before the burns, the main OMF isolated was unexpectedly Tulasnella calospora (Boud.) Juel. By 2014, after the burns, the expected Ceratobasidium sp. D.P. Rogers was the only OMF isolated in most burnt quadrats, whereas T. calospora was confined to a minority of unburnt ‘control’ and the ‘spring burn’ quadrats, which were also the only ones with flowering plants. The decline in rainfall during 2010–2012 probably caused the switch from mainly flowering to mainly vegetative plants and the change in OMFs. Burning in spring to summer was less damaging to this orchid than burning in autumn to winter, which should be noted by authorities in fire management plans for fire-prone areas in which this orchid occurs.

Clinacanthus nutans (Burm. f.) Lindau leaves are widely used by cancer patients and the leaf extracts possess cytotoxic and antiproliferative effects on several human cancer cell lines. However, the effect of C. nutans leaf extract on human melanoma, which is the least common but most fatal form of skin cancer and one of the most common cancers diagnosed in both sexes worldwide, is unknown. There is also limited information on whether the bioactivity of extracts differs between C. nutans leaves grown in different geographical locations with varying environmental conditions. The present study, for the first time, compared and demonstrated the cytotoxicity of the crude methanol extracts of C. nutans leaves from 11 different locations in Malaysia, Thailand and Vietnam, with diverse environmental conditions against D24 melanoma cells through WST-8 assay. The percentage of apoptotic cells following treatment with the most active extract was determined in a dose- and time-dependent manner by a cytofluorometric double staining technique. Biochemical and morphological changes in the treated and untreated cells were examined by fluorescence and transmission electron microscopy techniques, respectively, to further affirm the induction of apoptosis. The leaves of plants grown at higher elevations and lower air temperatures were more cytotoxic to the D24 melanoma cells than those grown at lower elevations and higher air temperatures, with the leaf extract from Chiang Dao, Chiang Mai, Thailand exhibited the highest cytotoxicity (24 h EC50: 0.95 mg/mL and 72 h EC50: 0.77 mg/mL). This most active crude extract induced apoptotic cell death in the D24 cells in a dose- and time-dependent manner. Typical biochemical and morphological characteristics of apoptosis were also observed in the treated D24 cells. The results, showing the cytotoxicity of C. nutans and the induction of apoptosis in D24 cells, are significant and useful to facilitate the development of C. nutans as a potential novel chemotherapeutic agent for the management of skin melanoma.

Background: Gac (Momordica cochinchinensis Spreng) seeds have long been used in traditional medicine as a remedy for numerous conditions due to a range of bioactive compounds. This study investigated the solvent extraction of compounds that could be responsible for antioxidant activity and anticancer potential. Methods: Defatted Gac seed kernel powder was extracted with different solvents: 100% water, 50% methanol:water, 70% ethanol:water, water saturated butanol, 100% methanol, and 100% ethanol. Trypsin inhibitors, saponins, phenolics, and antioxidant activity using the 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the ferric reducing antioxidant power (FRAP) assays; and anticancer potential against two melanoma cancer cell lines (MM418C1 and D24) were analysed to determine the best extraction solvents. Results: Water was best for extracting trypsin inhibitors (581.4 ± 18.5 mg trypsin/mg) and reducing the viability of MM418C1 and D24 melanoma cells (75.5 ± 1.3 and 66.9 ± 2.2%, respectively); the anticancer potential against the MM418C1 cells was highly correlated with trypsin inhibitors (r = 0.92, p < 0.05), but there was no correlation between anticancer potential and antioxidant activity. The water saturated butanol had the highest saponins (71.8 ± 4.31 mg aescin equivalents/g), phenolic compounds (20.4 ± 0.86 mg gallic acid equivalents/g), and antioxidant activity, but these measures were not related to anticancer potential. Conclusions: Water yielded a Gac seed extract, rich in trypsin inhibitors, which had high anticancer potential against two melanoma cell lines.