Analysis of the Antibacterial Activity and the Total Phenolic and Flavonoid Contents of the Moringa oleifera Leaf Extract as an Antimicrobial Agent against Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a bacterium that causes metal deterioration by forming biofilms on metal surfaces. This work was carried out to analyze the antibacterial activity and the phenolic and flavonoid contents of the Moringa oleifera leaf extract against Pseudomonas aeruginosa. M. oleifera leaves were extracted in a methanol solution at different concentrations. The M. oleifera leaf extract yields were 12.84%, 18.96%, and 19.64% for the 100%, 75%, and 50% methanol ratios, respectively. Extracts of M. oleifera leaves had a minimum inhibiting concentration (MIC) of approximately 6144 µg/mL against P. aeruginosa for a ratio of 100% methanol. In addition, no antibacterial activity was found for the 75% and 50% methanol ratios. The total phenolic levels were 16.26%, 12.73%, and 12.33% for the 100%, 75%, and 50% methanol solvent ratios, respectively. The total amounts of flavonoids were 23.32%, 3.40%, and 0.64% for the 100%, 75%, and 50% methanol solvents, respectively. The chemical structure of M. oleifera consists of kaemferol-3-O-rutinoside, quercimeritrin, kaempferol-3-O-ß-D-glucopyranoside, stearidonic acid, trichosanic acid, pyrophaeophorbide A, and stigmastan-3,6-dione. The concentration of the solvent is essential in the extraction of plant constituents. Different concentrations indicate differences in antibacterial activity, phenolic and flavonoid contents, and chemical structure.

Soursop leaves (Annona muricata L.) endophytic fungi anticancer activity against HeLa cells.

Cervical cancer causes many deaths in females worldwide, including in Indonesia. Several studies have reported that soursop (Annona muricata L.) leaves can be used to treat cervical cancer. This study aims to determine the use of endophytic fungi of A. muricata leaves extract as an ingredient that inhibits cervical cancer. The isolated endophytic fungi from various soursop leave accessions were grown in culture media, then extracted using ethyl acetate. The extract was then tested against anti-yeast, cervical cancer cells, and on normal cells as control using the MTT method. Five isolated fungi were selected based on the greatest inhibition in one concentration, and the inhibitory concentration 50 (IC50) value was determined. The soursop leaves endophytic fungi extracts showed cytotoxicity against cervical cancer cells by inhibiting the multiplication of HeLa cancer cells in vitro. The Sir-SM2 endophytic fungi crude ethyl acetate extract showed high cytotoxicity to cervical cancer cells (HeLa cells) but less harmful to the normal Chang cells; therefore can be a natural anticancer. Identification based on morphology shows that the isolated Sir-SM2 endophytic fungi belong to the Penicillium genus, and molecular identification based on Internal Transcribed Spacer shows high similarities with Penicillium crustosum.

Antimycobacterial activity of fruit of Zanthoxylum acanthopodium DC against M ycobacterium smegmatis.

OBJECTIVE: Fruits of lemon pepper (Zanthoxylum acanthopodium DC., Rutaceae) have been traditionally used as a spice and in folk medicine for treatment of diarrhea and stomachache. Stomachache could be associated with mycobacterial infection. The present study was designed to investigate the activity of Z. acanthopodium fruits against a non-infectious Mycobacterium smegmatis and to identify the important phytochemical constituent that is toxic towards mycobacteria. MATERIALS AND METHODS: The broth microdilution method was used to determine the minimal inhibitory concentration (MIC) of ethyl acetate or hexane extract of green, young fruits of Z. acanthopodium. Effect of active extract (hexane) on cell membranee integrity was studied by measuring sodium and potassium leakage into extracelullar liquid using Atomic Absorbtion Spectrophotometer (AAS). Next, cell morphology was observed by using Scanning Microscope Electron (SEM). Column chromatography was used for fractionation and purification of hexane extract while the chemical structure of the active compound was determined using NMR technique. Rifampicin, an antimycobacterial compound, was used as positive control. RESULTS: Hexane extract was active against M. smegmatis with an MIC of 64 µg/ml. Plant extract at the concentration of 128 µg/ml caused ions leakage. Concentration of sodium in extracellular liquid of cells treated with plant extract was significantly higher than that of untreated cells. SEM observation revealed cell wall deformation in cultures treated with the extract. NMR spectroscopy analysis of the most active fraction revealed that the compound that exerted toxicity on M. smegmatis was geranyl acetate. CONCLUSION: Geranyl acetate was an important constituent of Z. acanthopodium fruit that has antimycobacterial activity. Possibly, Z. acanthopodium fruit exert its toxic effects against M. smegmatis through damaging cell membrane.

Anticancer activity test of ethyl acetate extract of endophytic fungi isolated from soursop leaf (Annona muricata L.).

OBJECTIVE: To analyze anticancer activity of an ethyl acetate extract of endophytic fungi isolated from soursop leaf (Annona muricata L.). METHODS: Anticancer activity of fungal extracts was determined by observing its toxicity against MCF-7 (Michigan Cancer Foundation-7) cells in vitro by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay method. At an extract concentration of 100 µg/mL, 4 isolates out of 12 showed high activity against the cancer cell growth. The four isolates were then selected for further IC50 determination, by measuring the inhibition of cancer cell proliferation at extract concentration of 25 µg/mL, 50 µg/mL, 100 µg/mL, 200 µg/mL and 400 µg/mL. RESULTS: Results showed that isolate Sir-G5 had the highest anticancer activity with an IC50 of 19.20 µg/mL. The best isolates were screened again using a normal cell (Chang cells) to determine its toxicity against normal cells. Results indicated that the extracts do not affect the proliferation of normal cells. Molecular identification showed that the fungal isolate Sir-G5 has a close relationship with Phomopsis sp. CONCLUSIONS: The endophytic fungi isolated from soursop leaf has the potential to be used as a source of anticancer agents.

Screening and study of antifungal activity of leaf litter actinomycetes isolated from Ternate Island, Indonesia.

OBJECTIVE: To characterize abundance of leaf litter actinomycetes from Ternate Island and to assess the antifungal activity of actinomycetes isolates against Candida albicans, Saccharomyces cerevisiae (S. cerevisiae), and Aspergillus niger. METHODS: Actinomycetes were isolated from leaf litter of Durio species, Syzygium aromaticum, Piper betle, Myristica fragrans, or Pandanus species and unknown plants. Actinomycetes isolates were cultured in a liquid medium. Bioactive compounds were extracted and tested against fungal using Beury-Kirby method with modification. Minimum inhibitor concentration and cell leakages were conducted. Actinomycetes that produced the highest antifungal activity were indentified using molecular sequence data in 16S rRNA gene. RESULTS: Out of 50 selected isolates, two isolates MG-500-1-4 and SR-2-2 has highest activity against S. cerevisiae. Concentration of material containing nucleic acids, proteins, Ca(+) and K(+) ions and morphological observations indicated that extracts of MG-500-1-4 and SR-2-2 caused cell leakage and invagination of S. cerevisiae cells. Based on 16S rRNA gene identification, MG-500-1-4 and SR-2-2 isolates are similar to Streptomyces misakiensis and Streptomyces tricolor respectively. CONCLUSIONS: Ternate Island contains interesting biodiversity of actinomycetes that has potential use in agriculture, fisheries, and human health to reduce problem of fungal pathogen.

Natural product libraries to accelerate the high-throughput discovery of therapeutic leads.

A high-throughput (HT) paradigm generating LC-MS-UV-ELSD-based natural product libraries to discover compounds with new bioactivities and or molecular structures is presented. To validate this methodology, an extract of the Indo-Pacific marine sponge Cacospongia mycofijiensis was evaluated using assays involving cytoskeletal profiling, tumor cell lines, and parasites. Twelve known compounds were identified including latrunculins (1-4, 10), fijianolides (5, 8, 9), mycothiazole (11), aignopsanes (6, 7), and sacrotride A (13). Compounds 1-5 and 8-11 exhibited bioactivity not previously reported against the parasite T. brucei, while 11 showed selectivity for lymphoma (U937) tumor cell lines. Four new compounds were also discovered including aignopsanoic acid B (13), apo-latrunculin T (14), 20-methoxy-fijianolide A (15), and aignopsane ketal (16). Compounds 13 and 16 represent important derivatives of the aignopsane class, 14 exhibited inhibition of T. brucei without disrupting microfilament assembly, and 15 demonstrated modest microtubule-stabilizing effects. The use of removable well plate libraries to avoid false positives from extracts enriched with only one or two major metabolites is also discussed. Overall, these results highlight the advantages of applying modern methods in natural products-based research to accelerate the HT discovery of therapeutic leads and/or new molecular structures using LC-MS-UV-ELSD-based libraries.