Buah tanaman wualae (bahasa Tolaki) atau Etlingera elatior digunakan oleh masyarakat Tolaki (Sulawesi Tenggara) sebagai bumbu masak dan obat tradisional. Untuk mengungkap potensi tersebut, khususnya buah wualae, dilakukan kajian fitokimia dan farmakologi buah wualae. Kajian fitokimia meliputi isolasi dan penentuan struktur dikerjakan dengan metode kromatografi (kromatografi vakum cair dan kromatografi radial) dan spektroskopi (1H dan 13C NMR). Kajian farmakologi meliputi uji aktivitas biologi terhadap enam jenis bakteri (Bacillus subtilis FNCC 0060, Escherichia coli ATCC 35218, Pseudomonas aeruginosa ATCC 27853, Salmonella enterica ATCC 14028, Staphylococcus aureus ATCC 25923, dan Streptococcus mutans ATCC 25175). Hasilnya menunjukkan tiga senyawa fenolik telah berhasil diisolasi dan diidentifikasi dari buah wualae yaitu asam vanilat (1), asam p-hidroksibenzoat (2), dan asam kumarat (3). Ketiga senyawa tersebut pertama kali dilaporkan berhasil diisolasi dari buah E. elatior. Aktivitas antibakteri ekstrak metanol buah wualae dan tiga senyawa hasil isolasi menunjukkan bahwa sampel tersebut paling aktif terhadap bakteri S. mutans. Dengan demikian aktivitas senyawa-senyawa hasil isolasi dari buah E. elatior  berkorelasi dengan aktivitas ekstrak metanol buah E. elatior  terhadap S. mutans, dimana senyawa paling aktif adalah asam vanilat. Ekstrak yang dominan mengandung asam vanilat dari tanaman ini dapat dikembangkan sebagai herbal antibakteri S. mutans seperti herbal karies gigi dan obat kumur. Kata kunci: Etlingera elatior, wualae, buah, senyawa aromatik, antibakteri. The fruits of wualae (Tolakinese) or Etlingera elatior is utilized by Tolakinese (Sulawesi Tenggara) as spices and traditional medicine. To reveal the potencies, a study on phytochemical and pharmacological aspects have been carried out. The phytochemical study includes isolation and structure determination used chromatography methods (vacuum liquid chromatography and radial chromatography) and spectroscopy methods (1H NMR and 13C NMR). Biological activity was evaluated against some bacteria that are Bacillus subtilis FNCC 0060, Escherichia coli ATCC 35218, Pseudomonas aeruginosa ATCC 27853, Salmonella enterica ATCC 14028, Staphylococcus aureus ATCC 25923, and Streptococcus mutans ATCC 25175). The results showed that three aromatic compounds had been isolated and identified that are vanillic acid (1), p-hydroxybenzoic acid (2) and coumaric acid (3). All compounds are firstly reported that successfully isolated from fruits of Wualae. Antibacterial potency indicated that the methanol extracts and the isolated compounds are the most active towards S. mutans and have good activity correlation between the extracts and the compounds. Vanillic acid is the most active compound from E. elatior fruits. Thus, the extracts contain dominantly vanillic acid from          E. elatior fruits can be developed to be antibacterial herbals caused by S. mutans. Keywords: Etlingera elatior, wualae, fruits, aromatic compounds, antibacterial activity.

Flavonoids, present in fruits, vegetables and traditional medicinal plants, show anticancer effects in experimental systems and are reportedly non-toxic. This is a favorable property for long term strategies for the attenuation of lymph node metastasis, which may effectively improve the prognostic states in breast cancer. Hence, we studied two flavonoids, apigenin and luteolin exhibiting strong bio-activity in various test systems in cancer research and are readily available on the market. This study has further advanced the mechanistic understanding of breast cancer intravasation through the lymphatic barrier. Apigenin and luteolin were tested in a three-dimensional (3-D) assay consisting of MDA-MB231 breast cancer spheroids and immortalized lymph endothelial cell (LEC) monolayers. The 3-D model faithfully resembles the intravasation of breast cancer emboli through the lymphatic vasculature. Western blot analysis, intracellular Ca2+ determination, EROD assay and siRNA transfection revealed insights into mechanisms of intravasation as well as the anti-intravasative outcome of flavonoid action. Both flavonoids suppressed pro-intravasative trigger factors in MDA-MB231 breast cancer cells, specifically MMP1 expression and CYP1A1 activity. A pro-intravasative contribution of FAK expression in LECs was established as FAK supported the retraction of the LEC monolayer upon contact with cancer cells thereby enabling them to cross the endothelial barrier. As mechanistic basis, MMP1 caused the phosphorylation (activation) of FAK at Tyr397 in LECs. Apigenin and luteolin prevented MMP1-induced FAK activation, but not constitutive FAK phosphorylation. Luteolin, unlike apigenin, inhibited MMP1-induced Ca2+ release. Free intracellular Ca2+ is a central signal amplifier triggering LEC retraction through activation of the mobility protein MLC2, thereby enhancing intravasation. FAK activity and Ca2+ levels did not correlate. This implicates that the pro-intravasative contribution of FAK and of Ca2+ release in LECs was independent of each other and explains the better anti-intravasative effects of luteolin in vitro. In specific formulations, flavonoid concentrations causing significant anti-intravasative effects, can certainly be achieved in vivo. As the therapeutic strategy has to be based on permanent flavonoid treatment both the beneficial and adverse effects have to be investigated in future studies.

The rhizome of Etlingera elatior or Wualae (Tolakinese) has many advantages on traditional remedies and cooking in Sulawesi Tenggara. To support those advantages, two secondary metabolites derived from steroid and phenylpropanoic acid classes, stigmast-4-en-6β-ol-3-one (1) and p-coumaric acid (2), respectively, have been firstly isolated and identified from the E. elatior rhizome. Isolation of these two compounds was performed using several chromatography techniques, including thin layer chromatography (TLC), vacuum liquid chromatography (VLC) and radial chromatography (RC). Identification of isolates was carried out using 1H and 13C NMR spectroscopy and comparing the spectroscopy data with the library. The potency of antibacterial of the methanol extract of Wualae rhizome and the isolates were evaluated against Pseudomonas aeruginosa ATCC 27853, Bacillus subtilis FNCC 0060, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 35218, Salmonella enterica ATCC 14028, and Streptococcus mutans ATCC 25175 using agar diffusion method. Antioxidant activity was evaluated against DPPH radicals (2,2-diphenyl-1-picrylhydrazyl). The results show that the antibacterial potential of Wualae methanol extract is better than compound 1 and 2. Furthermore, the antioxidant properties of Compound 2 (IC50 159.47 μg/mL) was stronger than the antioxidant properties of Compound 1 (IC50 219.95 μg/mL) and the methanol extract (IC50 586.38 μg/mL).

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Metastasising breast cancer cells communicate with adjacent lymph endothelia, intravasate and disseminate through lymphatic routes, colonise lymph nodes and finally metastasize to distant organs. Thus, understanding and blocking intravasation may attenuate the metastatic cascade at an early step. As a trigger factor, which causes the retraction of lymph endothelial cells (LECs) and opens entry ports for tumour cell intravasation, MDA-MB231 breast cancer cells secrete the pro-metastatic arachidonic acid metabolite, 12S-hydroxy-5Z,8Z,10E,14Z-eicosatetraenoic acid [12(S)-HETE]. In the current study, treatment of LECs with 12(S)-HETE upregulated the expression of the transcription factors SRY-related HMG-box 18 (SOX18) and prospero homeobox protein 1 (PROX1), which determine endothelial development. Thus, whether they have a role in LEC retraction was determined using a validated intravasation assay, small interfering RNA mediated knockdown of gene expression, and mRNA and protein expression analyses. Specific inhibition of SOX18 or PROX1 significantly attenuated in vitro intravasation of MDA-MB231 spheroids through the LEC barrier and 12(S)-HETE-triggered signals were transduced by the high and low affinity receptors, 12(S)-HETE receptor and leukotriene B4 receptor 2. In addition, the current findings indicate that there is crosstalk between SOX18 and nuclear factor κ-light-chain-enhancer of activated B cells, which was demonstrated to contribute to MDA-MB231/lymph endothelial intravasation. The present data demonstrate that the endothelial-specific and lymph endothelial-specific transcription factors SOX18 and PROX1 contribute to LEC retraction.

Meistera chinensis is one of the new generations of the Zingiberaceae family. Zingiberaceae have a large number of species and still need research to reveal their chemical content. Meistera chinensis, a local plant that is widespread in Konawe Regency, Southeast Sulawesi. Empirically, M. chinensis is used as a flavor enhancer in food, aches, and increases body immunity. There are no reports of chemical content or its biological activity. In this study, the fruit was tested for phytochemicals, antioxidants using the ABTS test, total phenolic, total flavonoid content, and toxicity test with BSLT. Meistera chinensis was extracted with ethanol and fractionated by ethyl acetate solvent use vacuum liquid chromatography. Phytochemical screening was carried out qualitatively by using the calorimetric method. The fraction toxicity was monitored by a lethal test for brine shrimp (BSLT). These fractions for the ABTS method obtained fractions 1-8 (F1-F8) and ascorbic acid were used as controls. The results showed radical scavenging activities fraction of M. chinensis fruit was a very strong activity with IC50 of 42.7±3.53 mg/L (F8). The total phenolic and flavonoid contents were 30.72±1.07 mgGAE/g and 8.02±0.48 mgQE/g, respectively. The phytochemical evaluation contains terpenoids, saponins, phenolics, steroids, alkaloids, and flavonoids. The BSLT toxicity test was found to be very toxic with IC50 of 5.20±0.72 mg/L. These findings indicate that the fruit of M. chinensis acts as an antioxidant and toxicity agent.

Two known compounds, namely stigmast-4-en-6β-ol-3-one (1) and p-hydroxybenzoic acid (2) are firstly reported from stems of E. elatior. Isolation were carried out by chromatography technique including Thin Layer Chromatrography (TLC), vacuum liquid chromatography (VLC) and radial chromatography (RC) with silica gel as adsorbent and mixture of solvents as eluen. Structure of isolated compounds were determined by spectroscopy methods i.e. FTIR, 1H and 13C NMR and also by comparison the spectroscopies data with the same data from references. Biological activities of isolated compounds evaluated against some bacteria consist of Bacillus subtilis FNCC 0060, Escherichia coli ATCC 35218, Pseudomonas aeruginosa ATCC 27853, Salmonella enterica ATCC 14028, Staphylococcus aureus ATCC 25923, dan Streptococcus mutans ATCC 25175 and also against DPPH (2,2-diphenil 1-pichylhydazyl) radical as antiradical scavenger activity. The result showed that generally, antibacterial potency those samples i.e. methanol extract, stigmast-4-en-6β-ol-3-one and p-hydroxybenzoic acid less active than positive control (chloramphenicol). Individually, antibacterial activity of p-hydroxybenzoic acid > methanol extract > stigmast-4-en-6 β-ol-3-one. In addition, p-hydroxybenzoic acid is the most active against S. mutans. Radical scavenger activity of tested samples is less active than than positive control (ascorbic acid). Furthermore, the radical scavenger potency of p-hydroxybenzoic acid > methanol extract > stigmast-4-en-6 β-ol-3-one.

Soft coral Nepthea sp. grows in the seas of South-East Sulawesi, Indonesia. However, information on the chemical and pharmaceutical aspects of this genus is still limited. Therefore, this research aims to explore the chemical contents and biological activities of Nepthea sp. The sample was collected from the waters of Saponda Island by SCUBA diving. It was extracted by ethyl acetate and fractionated using vacuum liquid chromatography. The chemical content was analyzed by phytochemical screening, LC-MS/MS analysis, Total Phenolics Content and Total Flavonoids Contents. Antioxidant potency was evaluated by DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals and ABTS (2,2’-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid). Cytotoxicity property was analyzed by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays. The result showed that the fractionation of Nepthea ethylacetate extracts produced six fractions (A-F). Fractions A and B contain non-polar compounds. Based on LC-MS/MS data, the non-polar compounds in Fraction A and B include achillin, atractylenolide II, buthyl isobuthyl phthalate, rengyolester, 2a-acetoxycostic acid, ocotillol acetate, petasitolone and some unidentified compounds that are C33H58O4, C15H21NO, C21H33NO, and C16H20O4. In general, the antioxidant and cytotoxic properties of all samples are in the weak category, however, when examined for each sample, the antioxidant properties of fraction B is slightly better than fraction A based on the IC50 value of DPPH and ABTS. Cytotoxicity of Fraction A is better than Fraction B against Breast Cancer cell lines MCF-7. The non-polar fraction of Nepthea sp. can be developed as raw material for the discovery of new compounds, antioxidant and anticancer agents, especially breast cancer.