Anti-Acanthamoeba activity of a semi-synthetic mangostin derivative and its ability in removal of Acanthamoeba triangularis WU19001 on contact lens.

Garcinia mangostana L., also known as the mangosteen tree, is a native medicinal plant in Southeast Asia having a wide variety of pharmacologically active compounds, including xanthonoid mangostin. In this study, we examined the pharmacological activities of the selected semi-synthetic mangostin derivative, namely, amoebicidal activity, encystation inhibition, excystation activity, and removal capacity of adhesive Acanthamoeba from the surface of contact lens (CL). Among the three derivatives, C1 exhibited promising anti-Acanthamoeba activity against Acanthamoeba triangularis WU19001 trophozoites and cysts. SEM images displayed morphological changes in Acanthamoeba trophozoites, including the loss of acanthopodia, pore formation in the cell membrane, and membrane damage. In addition, the treated cyst was shrunken and adopted an irregular flat cyst shape. Under a fluorescence microscope, acridine orange and propidium iodide (AO/PI) staining revealed C1 induced condensation of cytoplasm and chromatin with the loss of cell volume in the treated trophozoites, while calcofluor white staining demonstrated the leakage of cell wall in treated cysts, leading to cell death. Interestingly, at the concentration ranges in which C1 showed the anti-Acanthamoeba effects (IC50 values ranging from 0.035-0.056 mg/mL), they were not toxic to Vero cells. C1 displayed the highest inhibitory effect on A. triangularis encystation at 1/16×MIC value (0.004 mg/mL). While C1 demonstrated the excystation activity at 1/128×MIC value with a high rate of 89.47%. Furthermore, C1 exhibited the removal capacity of adhesive Acanthamoeba from the surface of CL comparable with commercial multipurpose solutions (MPSs). Based on the results obtained, C1 may be a promising lead agent to develop a therapeutic for the treatment of Acanthamoeba infections and disinfectant solutions for CL.

Effects of Artonin E on Cell Growth Inhibition and Apoptosis Induction in Colon Cancer LoVo and HCT116 Cells.

Today, colon cancer is the leading cause of cancer death. In Thailand, colon cancer is the third most common cancer in men and the second in women. Currently, the treatments for colon cancer include surgery, chemotherapy, radiation therapy, immunotherapy, hormone therapy, targeted drug therapy, and stem cell therapy. However, some treatments have side effects for cancer patients, causing unwanted symptoms. In addition, targeted therapy comes with a high cost for patients. Therefore, bioactive compounds might be a good choice for colon cancer treatment. In this study, we investigated the effect of artonin E on apoptosis induction in colon cancer LoVo and HCT116 cells. The concentration ranges of artonin E at 3, 5, 10, and 30 µg/mL in LoVo cells and 1, 1.5, 2, and 3 µg/mL in HCT116 cells were examined. The results implied that artonin E decreased cell viability and increased apoptotic cells in a dose-dependent manner. In addition, artonin E stimulated mitochondrial membrane potential (ΔΨm) changes associated with apoptosis by increasing the sub-G1 population analyzed by flow cytometry. Western blotting showed that artonin E increased the proapoptotic protein, Bax, and decreased anti-apoptotic proteins' (Bcl-2 and Bcl-x) expression. Moreover, artonin E also increased cleaved caspase-7 and cleaved-PARP expression in both LoVo and HCT116 cells. Interestingly, artonin E induced apoptosis through p-ERK1/2, p-p38/p38, and p-c-Jun expression in both cells. Our results suggested that artonin E induced apoptosis via caspase activation associated with the MAPKs signaling pathway. Therefore, artonin E might be used as a potential anticancer drug for colon cancer in the future.

Phytochemical, anti-Acanthamoeba, and anti-adhesion properties of Garcinia mangostana flower as preventive contact lens solution.

Acanthamoeba keratitis infection extends due to the growing number of contact lens users. Indigenous plants including Garcinia mangostana play a vital role in human health and well being. Many species of this plant have been reported with myriads of potent medicinal properties. However, the aims of this study were, for the first time, to isolate compounds from the flower of G. mangostana and to test their anti-Acanthamoeba and anti-adhesion activity against Acanthamoeba triangularis. Powdered flowers of G. mangostana were extracted and chromatographed on a silica gel column. The structures of the compounds were established with the aid of 1H NMR. More so, the anti-Acanthamoeba and anti-adhesion properties were tested on a 96-well polystyrene microtiter plate and soft contact lenses. Scanning electron microscope (SEM) was used to determine the features of A. triangularis on contact lenses. Eight pure compounds were obtained, namely 9-hydroxycalabaxanthone, tovophillin A, garcinone E, garcinone B, α-mangostin, gartinin, 8-deoxygartinin and γ-mangostin. The extract and pure compounds exhibited anti-Acanthamoeba activity with MIC values in the range of 0.25-1 mg/mL. In addition, the extract and α-mangostin displayed significant activity against the adhesion of A. triangularis trophozoites both in polystyrene plate and in contact lenses at 0.5 × MIC (0.25 mg/mL). Furthermore, α-mangostin has the potential to remove A. triangularis adhesion in contact lenses similar to a commercial multipurpose solution (MPS). SEM study confirmed that crude extract and α-mangostin are effective as solutions for contact lenses, which removed A. triangularis trophozoites within 24 h. Alpha-mangostin was non-toxic to Vero cells at a concentration below 39 µM in 24 h. Crude extract of G. mangostana flower and its α-mangostin serve as candidate compounds in the treatment of Acanthamoeba infection or as lens care solution, since they can be used as a source of natural products against Acanthamoeba and virulence factor associated with the adhesion of A. triangularis.

Anti-Acanthamoeba synergistic effect of chlorhexidine and Garcinia mangostana extract or α-mangostin against Acanthamoeba triangularis trophozoite and cyst forms.

Acanthamoeba spp. can cause amoebic keratitis (AK). Chlorhexidine is effective for AK treatment as monotherapy, but with a relative failure on drug bioavailability in the deep corneal stroma. The combination of chlorhexidine and propamidine isethionate is recommended in the current AK treatment. However, the effectiveness of treatment depends on the parasite and virulence strains. This study aims to determine the potential of Garcinia mangostana pericarp extract and α-mangostin against Acanthamoeba triangularis, as well as the combination with chlorhexidine in the treatment of Acanthamoeba infection. The minimal inhibitory concentrations (MICs) of the extract and α-mangostin were assessed in trophozoites with 0.25 and 0.5 mg/mL, for cysts with 4 and 1 mg/mL, respectively. The MIC of the extract and α-mangostin inhibited the growth of A. triangularis trophozoites and cysts for up to 72 h. The extract and α-mangostin combined with chlorhexidine demonstrated good synergism, resulting in a reduction of 1/4-1/16 of the MIC. The SEM results showed that Acanthamoeba cells treated with a single drug and its combination caused damage to the cell membrane and irregular cell shapes. A good combination displayed by the extract or α-mangostin and chlorhexidine, described for the first time. Therefore, this approach is promising as an alternative method for the management of Acanthamoeba infection in the future.

Tedaniophorbasins A and B-Novel Fluorescent Pteridine Alkaloids Incorporating a Thiomorpholine from the Sponge Tedaniophorbas ceratosis.

Two new fluorescent pteridine alkaloids, tedaniophorbasins A (1) and B (2), together with the known alkaloid N-methyltryptamine, were isolated, through application of mass directed purification, from the sponge Tedaniophorbas ceratosis collected from northern New South Wales, Australia. The structures of tedaniophorbasins A and B were deduced from the analysis of 1D/2D NMR and MS data and through application of 13C NMR DFT calculations. Tedaniophorbasin A possesses a novel 2-imino-1,3-dimethyl-2,3,7,8-tetrahydro-1H-[1,4]thiazino[3,2-g]pteridin-4(6H)-one skeleton, while tedaniophorbasin B is its 2-oxo derivative. The compounds show significant Stokes shifts (~14,000 cm-1) between excitation and emission wavelengths in their fluorescence spectra. The new compounds were tested for bioactivity against chloroquine-sensitive and chloroquine-resistant strains of the malaria parasite Plasmodium falciparum, breast and pancreatic cancer cell lines, and the protozoan parasite Trypanosoma brucei brucei but were inactive against all targets at 40 µM.

New flavonoids and xanthone from the stem bark of Artocarpus rigidus blume and cytotoxicity.

Three new flavonoids named artorigidinones A-C and a new xanthone named artorixanthone together with seven known compounds were isolated from the stem bark of Artocarpus rigidus Blume. Their structures were characterized by spectroscopic data. γ-Geranylapigenin exhibited cytotoxicity to a fibroblast-like cell line (SW1353) (IC50 < 0.32 µg/mL) stronger than a standard drug.

Artonin E sensitizes TRAIL-induced apoptosis by DR5 upregulation and cFLIP downregulation in TRAIL-refractory colorectal cancer LoVo cells.

BACKGROUND: The TRAIL treatment is an ideal strategy for colorectal cancer (CRC) therapy because of minimal collateral damage to normal cells. Unfortunately, some CRC is TRAIL-refractory cancer, such as LoVo cells. In an effort to overcome TRAIL-refractory cancer, we investigated the effect of artonin E in regulating death receptor 5 (DR5) and cellular FLICE (FADD-like IL-1ß-converting enzyme)-inhibitory protein (cFLIP), two major mediators regulate TRAIL-induced apoptosis, in LoVo cells as a model of TRAIL refractory CRC. METHODS: TRAIL-refractory cancer (LoVo cells) was treated with artonin E and TRAIL. Cell viability was determined by MTT assay. Apoptotic chromatin condensation was observed by fluorescent Hoechst33342 staining. The mRNA and protein expression of DR5 and FLIP was determined by quantitative PCR and Western blotting analysis, respectively. RESULTS: The combination treatment of artonin E and TRAIL enhanced cytotoxicity and apoptotic chromatin condensation in LoVo cells significantly, while treatment of artonin E or TRAIL alone was not. Artonin E enhanced both mRNA and protein expression of DR5. Interestingly, this is the first report showing that artonin E decreased protein expression of cFLIP. All together we showed that artonin E enhanced TRAIL-induced apoptosis in LoVo cells through DR5 upregulation and cFLIP downregulation. CONCLUSIONS: Artonin E was able to increase DR5 expression and decrease cFLIP expression in LoVo cells. These results showed that LoVo cells sensitized TRAIL-induced apoptosis in combined treatment with artonin E and TRAIL. Therefore, the combination treatment of artonin E and TRAIL is one of the potential strategies used for TRAIL-refractory CRC therapy in the future.

Goniothalamin induces mitochondria-mediated apoptosis associated with endoplasmic reticulum stress-induced activation of JNK in HeLa cells.

Goniothalamin, a natural occurring styryl-lactone isolated from Goniothalamus macrophyllus (Blume) Hook. f. & Thomson var. macrophyllus, can trigger cancer cell death in various types of cancer cell. The present study focused on elucidation of the mitochondria-mediated apoptosis associated with endoplasmic reticulum (ER) stress-induced activation of c-Jun NH2-terminal kinase (JNK) by goniothalamin in HeLa cervical cancer cells. Cell viability was determined using an MTT assay, and DNA condensation and loss of mitochondrial membrane potential were determined using Hoechst 33342 and JC-1 staining, respectively. Flow cytometry was used for cell cycle and phosphatidyl-serine exposure analyses. Apoptotic-associated ER stress signaling pathways were determined using immunoblotting, reverse transcription-polymerase chain reaction (RT-PCR) and RT-quantitative PCR analyses. The results suggested that goniothalamin suppressed cell proliferation in a time- and dose-dependent manner. The induction of apoptosis was confirmed by increased DNA condensation, loss of mitochondrial membrane potential and cell surface phosphatidyl-serine presentation. The cell cycle analysis demonstrated that the goniothalamin-treated HeLa cells were in G2/M arrest. Determination of the caspase cascade and apoptotic proteins indicated the induction of apoptosis through the intrinsic pathway. In addition, the levels of phosphorylated JNK and the transcription factor, C/EBP homologous protein (CHOP), an ER stress-associated apoptotic molecule, were increased in the goniothalamin-treated cells. These data indicated that goniothalamin exerted a cytotoxic effect against HeLa cells via the induction of mitochondria-mediated apoptosis, associated with ER stress-induced activation of JNK.

Xanthones from the green branch of Garcinia dulcis.

Two new prenylated xanthones, namely dulcisxanthone H and dulcisxanthone I along with garciniaxanthone C, were isolated from the dichloromethane extract of the green branch of Garcinia dulcis. Their structures were elucidated by the analysis of 1-D and 2-D NMR spectral data. Their antibacterial activities were also examined.

Constituents of Fagraea fragrans with Antimycobacterial Activity in Combination with Erythromycin.

Seven new compounds constituted by three secoiridoids (1-3), two isocoumarins (4 and 5), an iridoid (6), and an aromatic derivative (7) in addition to 24 known compounds were isolated from the stem bark of Fagraea fragrans. The structures of the new compounds were determined on the basis of spectroscopic data analysis. The isolated compounds showed no antibacterial activity against Staphylococcus aureus and Escherichia coli. However, 5-formylisochromen-1-one (4), (-)-mellein (8), and swermacrolactone C (9) exhibited potent antimycobacterial activities against Mycobacterium smegmatis when used in combination with the antibiotic drug erythromycin.

Anti-Proliferation and Apoptosis Induction in Breast Cancer Cells by Cratoxylum cochinchinense Extract.

Background: Breast cancer is the most common invasive cancer in females worldwide. It was found about 37.5% in Thai females and is one of the leading causes of death-related cancers in women. Therefore, new finding of anti-cancer compound as a therapeutic candidate in breast cancer is necessary. Objective: To investigate the effect of Cratoxylum cochinchinense extract on anti-proliferation and apoptosis induction in breast cancer cells. Material and Method: Cell proliferation and cell viability assay were determined by MTT assay. Hoechst 33342 and JC-1 staining were used to determined nuclear morphological changes and mitochondrial membrane potential, respectively. Resu;ts: C. cochinchinense extract showed anti-proliferation in MDA-MB-468 treated cells in a time- and dose-dependent manner with IC50 value of 19.19+0.8 µg/ml. In addition, C. cochinchinense extract induced nuclear condensation and apoptotic bodies in MDA-MB-468 treated cells. JC-1 staining revealed that C. cochinchinense extract induced mitochondrial membrane dysfunction. Conclusion: C. cochinchinense extract showed anti-proliferation and apoptosis induction properties in MDA-MB-468 treated cells. These results suggested that C. cochinchinense extract may be a potential candidate for anti-cancer drug developing. The underlying mechanisms of apoptosis induction should be further studied.

Emilia sonchifolia extract activity against white spot syndrome virus and yellow head virus in shrimp cell cultures.

Emilia sonchifolia (L.) DC is a plant used in traditional medicine to treat several viral and bacterial diseases. The antiviral activities of selected Sephadex LH-20 column fractions and HPLC subfractions of an acetone extract of E. sonchifolia leaves were determined in shrimp Penaeus merguiensis primary lymphoid cells infected with either white spot syndrome virus (WSSV) or yellow head virus (YHV). WSSV and YHV replication was quantified using quantitative real-time PCR tests targeted to the VP19 and ORF1b gene transcripts, respectively. In lymphoid organ cells exposed to 100 µg ml⁻¹ of either the Sephadex fraction F14 or the HPLC F14 subfraction SF4, both fractions caused reduced replication, but YHV replication was reduced only by SF4. In the asthiazolyl blue mitochondrial enzyme activity assays to assess extract cytotoxicity, >60% of primary lymphoid organ cells remained viable following exposure to 100 µg ml⁻¹ of either F14 or SF4. GC-MS analysis of the HPLC F14 subfraction SF4 showed that it contained 2,4-di-tert-butylphenol. This study is the first to show that E. sonchifolia leaf extracts might be useful as bioactive agents to protect shrimp against viruses such as WSSV and YHV.

A new anthraquinone from Morinda elliptica Ridl.

A new anthraquinone, morinquinone, together with 18 known anthraquinones were isolated from the stems of Morinda elliptica Ridl. Their structures were elucidated on the basis of spectroscopic data. They each showed weak inhibitory activity against a susceptible strain of Staphylococcus aureus and a methicillin-resistant S. aureus. Damnacanthal was effective against Microsporum gypseum (MIC 1 µg/mL). Lucidin was active against Entamoeba histolytica (MIC 31.25 µg/mL) and Giardia intestinalis (MIC 7.8 µg/mL).

A new prenylated biflavonoid from the leaves of Garcinia dulcis.

A new prenylated biflavonoid, named dulcisbiflavonoid A, together with five biflavonoids were isolated from the leaves of Garcinia dulcis. Their structures were elucidated by analysing their spectroscopic data, especially 1D and 2D NMR.

Antiproliferation and Apoptosis Induction in Colorectal Cancer Cells by Goniothalamin.

OBJECTIVE: To investigate the effect of goniothalamin on antiproliferation and apoptosis induction in three types of colorectal cancer cells. BACKGROUND: Colorectal cancer is the third of the twentieth most commonly diagnosed cancer. Different types of colorectal cancer cells differ in genotype and characteristics leading to different responses to anticancer drugs. Therefore, finding new anticancer compound for the colorectal cancer cells is necessary. MATERIAL AND METHOD: Antiproliferative response of goniothalamin on three colorectal cancer cell lines including Colo 205, SW480, and LoVo were determined by MTT assay. The antiproliferative response at different time and dose was also observed. Apoptosis induction by goniothalamin was observed in all three cell-lines via morphological changes and nuclear condensation by Hoechst33342 staining. RESULTS: Goniothalamin showed different antiproliferative response on Colo 205, SW480, and Lo Vo cells at the IC50 value is 9.86 ± 0.38 µM, 22.00 ± 4.40 µM, and 65.25 ± 1.85 µM respectively. In addition, the antiproliferative response of goniothalamin was a time- and dose-dependent manner Apoptosis morphological changes and nuclear condensation were clearly observed in Colo 205, SW480 and LoVo cells treated with 10 µM, 25 µM, and 50 µM goniothalamin, respectively. CONCLUSION: Goniothalamin showed antiproliferation and apoptosis induction in colorectal cancer cells with different sensitivity depending on cell type. Investigation of mechanisms underlying apoptosis and its potential use for colorectal cancer treatment should be further studied.

New depside from Citrus reticulata Blanco.

A new depside, named depcitrus A (1), and 31 known compounds were isolated from the peels, leaves and branch barks of Citrus reticulata Blanco. Methylation of the high polarity fractions from the branch barks and peels gave one new methylated compound named depcitrus B (14) and five known compounds. Their structures were established based on spectroscopic evidence. The antioxidant, antimicrobial and cytotoxic activities of some pure compounds were evaluated.

A new bisanthraquinone and cytotoxic xanthones from Cratoxylum cochinchinense.

A new bisanthraquinone has been isolated from the stems of Cratoxylum cochinchinense together with vismiaquinone C and 16 known xanthones. Their structures were characterised by using spectroscopic methods. Two of the isolated compounds are modified xanthones (6 and 15) and exhibited strong cytotoxicity against human epidermoid carcinoma (A341: IC50 2.01, 1.78 µg/mL) and human breast cancer cell lines (SKBR3: IC50 1.54, 0.69 µg/mL).

New acridone from the wood of Citrus reticulata Blanco.

A new acridone, named citruscridone (1) together with five known compounds were isolated from the wood of Citrus reticulata Blanco. Their structures were established based on spectroscopic evidence. The antibacterial and antifungal activities of the wood extracts and pure compounds were evaluated.

A new chromone from the leaves of Melaleuca cajuputi Powell.

A new chromone, 5,7-dihydroxy-2 -(hydroxymethyl)-6,8-dimethyl-chromen-4-one, named melachromone, together with 12 known compounds, including chromones, anthraquinone, flavonoids, flavonoid glycosides, benzene derivatives, ellagic acids and terpenes, were isolated from the leaves of Melaleuca cajuputi Powell. Their structures were characterised by spectroscopic methods.

Alkaloid and coumarins from the green fruits of Aegle marmelos.

Five (1-5) and 15 known compounds were isolated from the acetone extract of the green fruits of Aegle marmelos. The structure of compounds 1-5, marmesiline (1), 6-(4-acetoxy-3-methyl-2-butenyl)-7-hydroxycoumarin (2), 6-(2-hydroxy-3-hydroxymethyl-3-butenyl)-7-hydroxycoumarin (3), marmelonine (4) and 8-hydroxysmyrindiol (5), were determined on the basis of spectroscopic analyses. Antifungal and antibacterial activities of selected compounds were also evaluated.