Taxonomy and Antimicrobial Activity of Gliding Bacterium from Indonesian Mangroves

The discovery of new antibiotic is needed, due to the increasing antimicrobial resistance in nature. Therefore, this study aims to isolate gliding bacteria and to ascertain their antimicrobial activity against pathogenic microbes. This research was conducted by isolating gliding bacteria from mangrove sediment in Muara Angke, North Jakarta, Indonesia. All of the strains were identified by 16S rRNA genes sequencing and those selected strain was characterized using polyphasic approach. The performance of crude extracts against to ten pathogenic microorganisms were detected using serial dilution test in 96-well plates. Mangrove gliding bacteria isolates designated 313MSO and 314MSO were showed high homology to Ohtaekwangia kribbensis with 96.20% and 99.12% similarity, respectively. The polyphasic approach led to the conclusion that the strain 313MSO was a new species of the genus Ohtaekwangia. Twenty-three crude extracts were obtained from cultivating the strain in twenty-three different media. The most of them inhibited the growth of Staphylococcus aureus Newman with the minimum concentration of 33.33-66.67 µg/ml. Four compounds (Marinoquinolines A-D) were obtained from HPLC-MS analysis. Furthermore, the strain 313MSO is presently being studied for in-depth identification of additional unknown metabolites detected in the crude extracts

REGULATION OF ADIPOGENESIS AND KEY ADIPOGENIC GENE EXPRESSION BY MANGOSTEEN PERICARP EXTRACT AND XANTHONES IN 3T3-L1 CELLS

Obesity is one of the risk factors for atherosclerosis and its fat occurrence and development are associated with fat accumulation and adipocyte differentiation.  Thus, the suppression of adipocyte differentiation can be a potential anti-obesity approach to this health concern. This study examined the effect of mangosteen pericarp extract (MPE) and xanthone (α-Mangostin (AM) and γ-Mangostin (GM)) on the expression of PPARγ, C/EBPα, SCD1, LPL, aP2, adipoQ, and FAS in 3T3-L1 cells. Concentrations of MPE and xanthones used were based on the cytotoxic assay on 3T3-L1 cells. Three different MPE concentrations (0, 25, and 50 µg/ml) and three different AM concentrations (0, 25 and 50 µM) and GM (0, 50, and 75 µM) were used in the experiment. The expressions of PPARγ, C/EBPα, SCD1, LPL, aP2, adipoQ, and FAS genes were measured using real-time quantitative PCR. The expression of the genes was down-regulated in the group of cells treated with 50 µg/ml of MPE and 50 µM of GM. However, the 25 µM and 50 µM of AM did not suppress PPARγ and SCD-1 expression. The 50 µM of AM also failed to reduce aP2 gene expression. Finally, MPE and GM showed potential anti-adipogenesis and anti-obesity effects by suppressing the expression of PPARγ, C/EBPα, SCD1, LPL, aP2, adipoQ and FAS genes in 3T3-L1 cells

Standardization of Pegagan Extract, Centella Asiatica as Hepatoprotectiveherbal Medicine

Herbal medicinal products would be affected by the quality of raw materials. In turn, the quality of raw material will also be influenced by various factors such as soil conditions, cultivation, post-harvest processing, and the processing of raw materials into crude drug or extract. Therefore, in order to make good herbal medicines, it is necessary to make standardization of herbal extracts that produced herbal medicines that have the same quality and functions of effectiveness in each process. From preliminary studies that have been done, Centella asiatica is one of the potential plants as a source of hepatoprotective compounds. Test in vivo and in vitro against Centella asiatica extracts have shown very good results. Ethyl acetate extract with 17.5 mg/kg of doses body weight and butanol 228.8 mg/kgof doses body weight has been applied for in vivo test using mice induced by CCl4; theydemonstrated hepatoprotective effects. Ethyl acetate extracts were able to reduce levels of the enzyme alanine aminotransferase (ALT) and aspartate aminotransferase (AST) by 56 % and 44 % respectively while butanol extract can reduce the enzymes AST levels by 3%. Standardizationof Centella asiatica extract performed in this study was the characterization of the extract in the form of non-specific and specific parameters corresponding to the reference of PPOMN (Ministry of health Republic of Indonesia, 2000) such as levels of drying shrinkage, ash content, total plate count microbial contamination, levels of water-soluble compounds, levels of compounds that are soluble in ethanol, phytochemical test, total phenolic content, total flavonoid content and the determination of Pb and Cd weight.The results showed that non-specific parameters for the ethanol extract of Centella asiatica were requirements based on Herbal Pharmacopoeia in 2008 which includes parameters such as determination of shrinkage on drying ≤ 10%, ash content ≤ 16.6% and negative microbial contamination. Specific parameters for the ethanol extract of Centella asiatica have met the requirements of Herbal pharmacopeia in 2008

Cholestan Steroids from The Stem Bark of Aglaia angustifolia Miq and Their Cytotoxic Activity against MCF-7 Breast Cancer Cell Lines

With about 120 species, Aglaia is one of the largest genera of the plant family Meliaceae (the mahogany plants). Various Aglaia species have been investigated since the 1960s for their phytochemical constituents and biological properties. This research objective was to find secondary metabolites that have activity as anti-breast cancer compounds from endemic Indonesian Aglaia, such as Aglaia angustifolia Miq. Two cholestan type steroids, stigmast-5en-3α-acetat (1),  as a new steroid with α-sterochemistry of acetyl moiety at C-3 and 23a-homostigmast-5en-3β-ol (2), with unusual side chain were isolated for the first time from the stem bark of Aglaia angustifolia Miq or known as segara tree in Kalimantan. The chemical structures of two steroids were identified with spectroscopic data, including IR, NMR (1H, 13C, DEPT 135°, HMQC, HMBC, NOESY, 1H-1H COSY) and HRTOF-MS, as well as by comparing with previously reported spectral data. These two steroids were isolated for the first time from this genus. Steroids 1 and 2 were evaluated for cytotoxic activity against MCF-7 breast cancer cells and showed weak activity with IC50 values of 829.0 and 903.0 µg/mL, respectively

Induction of Matrix Metalloproteinases in Chondrocytes by Interleukin IL-1β as an Osteoarthritis Model

Osteoarthritis (OA) is a chronic disease of the joints and bones due to trauma or other joint-related diseases (secondary). Synovial inflammation commonly causes disturbance in joint homeostasis, which is associated with OA. Enzymes such as aggrecanase and metalloproteinase generate cartilage damage, mediated by tumor necrosis factor (TNF-α) and interleukin (IL)-1. Pro-inflammatory cytokines, including TNF-α, IL-1β, and IL-6, are responsible for regulation of the extracellular matrix, cartilage degradation, and apoptosis of chondrocytes. This study aimed to observe the cell viability and expression level of matrix metalloproteinases (MMP-1 and MMP-3) and tissue inhibitor metalloproteinases (TIMP-1 and TIMP-2) in human chondrocyte cells (CHON-002) induced by IL-1β. CHON-002 was induced with IL-1β (0.1, 1 and 10 ng/mL) as an OA model. The viability of the cells was measured with a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxyme-thoxyphenyl)-2-(4-sulfophenyl)-2H-tetra zolium (MTS) assay, while expression of MMP-1, MMP-3, TIMP-1, and TIMP-2, was evaluated by RT-PCR. The viability of IL-1β-induced CHON-002 (CHON-002- IL-1β) cells at day 1 and 5 showed that treatment with up to 10 ng/mL of IL-1β was not toxic. Expression of TIMP-1 and TIMP-2 in CHON-002-IL-1β was lower compared to control, while that of MMP-1 and MMP-3 was higher compared to control. These results indicate that CHON-002 treated with 10 ng/mL IL-1β has expression patterns consistent with chondrocyte damage, so the CHON-002-IL-1β system may serve as a model for MMP induction in OA

Effects of insulin-like growth factor-induced Wharton jelly mesenchymal stem cells toward chondrogenesis in an osteoarthritis model

Objective(s): This study aimed to determine the collagen type II (COL2) and SOX9 expression in interleukin growth factor (IGF-1)-induced Wharton’s Jelly mesenchymal stem cells (WJMSCs) and the level of chondrogenic markers in co-culture IGF1-WJMSCs and IL1β-CHON002 as osteoarthritis (OA) cells model. Materials and Methods: WJMSCs were induced with IGF1 (75, 150, and 300 ng/ml) to enhance their chondrogenesis capability. The gene expression of SOX9 and COL2 was evaluated with quantitative RT-PCR. Furthermore, IGF1-WJMSCs were co-cultured with IL1β-CHON002 cells in varied ratios (1:2, 1:1, 2:1). Chondrogenic markers ADAMTS1, ADAMTS5, MMP3, MMP1, and RANKL were measured with ELISA. Results: The IGF1-WJMSCs had an increased expression of COL2 and SOX9. ADAMTS1, ADAMTS5, MMP1, MMP3, and RANKL levels were decreased in the co-culture IGF1-WJMSCs and IL1β-CHON002. Conclusion: The IGF1-induced WJMSCs were capable to enhance chondrogenesis, indicated by increased expression of SOX9 and COL2 and decreased expression of ADAMTS1, ADAMTS5, MMP3, MMP1, and RANKL. These findings can be further used in the osteoarthritis treatment

Effect of interleukins (IL-2, IL-15, IL-18) on receptors activation and cytotoxic activity of natural killer cells in breast cancer cell

Introduction: Breast cancer is one of the leading cause of cancer deaths in women. Metastasis in BC is caused by immunosurveillance deficiency, such NK cell maturation, low NK activity and decreasing cytotoxicity. This study was performed to improve activating receptors and cytotoxicity of NK cells using interleukins (ILs). Methods: Human recombinant IL-2, -15, and -18 were used to induce NK cells. We measured the activating and inhibiting receptors, proliferation activity of NK cells, and the cytotoxicity of NK cells on BC cells (MCF7). The effects of ILs were tested on the NK cell receptors CD314, CD158a and CD107a with flowcytometry, proliferation at various incubation times with 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy methoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and concentrations of TNF-\u3b1 and IFN-\u3b3 by NK cells with ELISA. Results: ILs increased NK cell receptor levels (CD314, CD158a, and CD107a) at 24 hours of incubation. ILs increased NK cell viability, which increased with longer incubation. Moreover, ILs-induced NK cells inhibited proliferation in MCF7 cells, as well as increased TNF-\u3b1, IFN-\u3b3, PRF1 and GzmB secretion. Conclusion: IL-2, IL-15, and IL-18 improved activating receptors and proliferation of NK cells. IL-induced NK cells increased TNF-\u3b1, IFN-\u3b3, PRF1 and GzmB secretion and cytotoxic activity on BC cells. High NK cell numbers increased BC cell growth inhibition

Biosynthesis of Polyketides in <i>Streptomyces</i>

Polyketides are a large group of secondary metabolites that have notable variety in their structure and function. Polyketides exhibit a wide range of bioactivities such as antibacterial, antifungal, anticancer, antiviral, immune-suppressing, anti-cholesterol, and anti-inflammatory activity. Naturally, they are found in bacteria, fungi, plants, protists, insects, mollusks, and sponges. Streptomyces is a genus of Gram-positive bacteria that has a filamentous form like fungi. This genus is best known as one of the polyketides producers. Some examples of polyketides produced by Streptomyces are rapamycin, oleandomycin, actinorhodin, daunorubicin, and caprazamycin. Biosynthesis of polyketides involves a group of enzyme activities called polyketide synthases (PKSs). There are three types of PKSs (type I, type II, and type III) in Streptomyces responsible for producing polyketides. This paper focuses on the biosynthesis of polyketides in Streptomyces with three structurally-different types of PKSs

Biosynthesis of Polyketides in <i>Streptomyces</i>

Polyketides are a large group of secondary metabolites that have notable variety in their structure and function. Polyketides exhibit a wide range of bioactivities such as antibacterial, antifungal, anticancer, antiviral, immune-suppressing, anti-cholesterol, and anti-inflammatory activity. Naturally, they are found in bacteria, fungi, plants, protists, insects, mollusks, and sponges. Streptomyces is a genus of Gram-positive bacteria that has a filamentous form like fungi. This genus is best known as one of the polyketides producers. Some examples of polyketides produced by Streptomyces are rapamycin, oleandomycin, actinorhodin, daunorubicin, and caprazamycin. Biosynthesis of polyketides involves a group of enzyme activities called polyketide synthases (PKSs). There are three types of PKSs (type I, type II, and type III) in Streptomyces responsible for producing polyketides. This paper focuses on the biosynthesis of polyketides in Streptomyces with three structurally-different types of PKSs