Endometriosis prevalence has been known to be quite high among women of reproductive age and with... more Endometriosis prevalence has been known to be quite high among women of reproductive age and with pelvic pain and/or infertility. The reason is that the estrogen level in the eutopic endometrium of women with endometriosis is higher than in normal endometrium which may possibly be caused by the lack of interaction between progesterone and progesterone receptor (PR). Dexa Laboratories of Biomolecular Sciences (DLBS) has developed DLBS1442, a bioactive fraction from Phaleria macrocarpa (Scheff) Boerl fruit, which has been found to be potential to treat symptoms of primary dysmenorrhea and alleviate endometriosis. Therefore, the identification of DLBS1442 active compounds which act as a PR agonist was necessary. Identification was performed using metabolomics study which resulted in 14 compounds. Crystal structure of the PR with asoprisnil as the reference was obtained from PDB (4A2J). Virtual screening validation process was performed using Protein-Ligand ANT System (PLANTS) and Python-based Protein-Ligand Interaction Fingerprinting (PyPLIF). According to the virtual screening protocol validation, the highest Enrichment Factor (EF) 1% value was obtained with hydrogen interaction with GLN725 and ARG766 residue. Virtual screening of the DLBS1442 metabolomics study showed that only glyceryl pentacosanoate exhibited a lower Chem Piecewise Linear Potential (ChemPLP) than the cutoff. This compound might have a role as a PR agonist which supported the previous findings of DLBS1442 to alleviate endometriosis. However, this finding requires further in vitro and/or in vivo study to ensure the agonist activity of glyceryl pentacosanoate as a DLBS1442 active compound.
Inflammation is characterized by an increase in vascular permeability leading to edema formation.... more Inflammation is characterized by an increase in vascular permeability leading to edema formation. Phosphoinositide 3-kinase (PI3K) signaling pathway plays the main role to regulate several key events in the inflammatory response to damage and infection. Our previous in silico study showed that anthocyanin and ternatin flavonoids may offer anti-inflammatory potential via molecular docking model. Here, we explored whether Clitoria ternatea anthocyanin extract (CTA) had protective activity against inflammation via regulating gene expression related to PI3Ks in rat edema. CTA was extracted in methanol and acetone using maceration and lyophilization. Edema model was done by injecting carrageenan into rat paw. Oral supplementation of CTA (100, 250, and 500 mg/kg BW) and ibuprofen standard (15 mg/kg BW) was given to rats daily for 7 days. After sample treatment, the right hind paw of the rat was injected with carrageenan to induce edema condition. The ability of CTA to suppress the inflammation in paw edema was measured by quantifying the increased percentage of rat paw volume at 1, 3, and 5 hrs after injection. Rat paw tissue was collected, and gene expression related to PI3Ks, i.e. Akt1, PKB, Ilk, Pdk2, Pik3Ca, and p53 was determined using quantitative PCR (qPCR). Carrageenan showed a time-dependent increase in volume of rat paw starting from injection until the next 5 hrs. It reached maximum volume increase of rat paw at 3 hrs after injection. CTA treatment had a dose-dependent pattern in reducing the edema volume in carrageenan-induced paw edema. At 500 mg/kg BW, CTA significantly suppressed the inflammation in paw edema compared to that of positive control, indicating by the low percentage of paw edema volume at 1, 3, and 5 hrs after carrageenan injection. At gene level, qPCR data showed that CTA significantly down-regulated the expression of genes involved in PI3Ks, such as Akt1, PKB, Ilk, Pdk2, Pik3Ca, and p53 in paw tissues. Thus, CTA may have a protective effect against edema via suppressing genes of PI3Ks.
Butterfly pea (Clitoria ternatea) is an indigenous plant from Moluccas in Indonesia. Due to its p... more Butterfly pea (Clitoria ternatea) is an indigenous plant from Moluccas in Indonesia. Due to its pharmacological properties, the flower of this plant is also used as a medicine in Ayurvedic practices in India. The present work was aimed to determine the antidiabetic activity of C. ternatea protein extract (PCT) via the inhibition of α-amylase activity in vitro, and the modulation of genes related to diabetes in diabetic mice in vivo. C. ternatea flower protein extract was prepared using isoprecipitation, and its efficacy as an antidiabetic candidate was tested in vitro through α-amylase inhibition assay, and in vivo through the measurement of blood glucose level and diabetes-related genes expression, including those that encode for peroxisome prolif-erator-activated receptors gamma (PPARγ), glucose transporter 2 (Glut2), transcription factor 7-like 2 (Tcf7l2), calpain-10 (Capn10), and monocyte chemoattractant protein 1 (MCP1) in alloxan-induced diabetic mice. PCT showed α-amylase inhibition in a dose-dependent manner. The highest α-amylase inhibition (20.63%) was observed at 1 mg/mL of PCT. The in vivo study showed that PCT decreased the blood glucose level in alloxan-induced diabetic mice. PCT also altered the expression of diabetes-related genes. In adipose and skeletal muscle tissues, PCT upregulated the gene expression of PPARγ and Tcf7l2, meanwhile MCP1 expression was downregulated. In the pancreas, PCT upregulated the gene expression of Glut2, Capn10, and Tcf7l2. These results indicate that PCT is a potential antidiabetic agent for the treatment of type 2 diabetes.
Inflammatory markers such as cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), myel... more Inflammatory markers such as cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), myeloperoxidase (MPO), and prostaglandin (PEG) are widely known as major targets in discovering natural anti-inflammatory drugs for the treatment of inflammation-related diseases. Non-steroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen and aspirin are mostly used at present, however, some NSAIDS have been reported to cause gastrointestinal side effect due to ligand-protein interaction. Molecular docking is a promising tool to study such modes of interaction. In this study, we evaluated the potential use of anthocyanin and ternatin flavonoids as natural anti-inflammatory agents for treatment of inflammatory-related diseases using in silico molecular docking assay. Automated docking study using Protein-Ligand ANT System (PLANTS) and AutoDock Vina was performed with various ligand molecules, including ibuprofen, anthocyanin, and ternatin against the protein crystal structures of COX-1, COX-2, iNOS, and MPO. The in silico data demonstrated that ibuprofen bound effectively to the active site of COX-1 and MPO with minimum binding energy, yet the compound required more energy to bind the active site of COX-2. Ternatin flavonoid was bound to COX-2 and iNOS with minimum binding energy. In terms of binding energy, anthocyanin flavonoid was found to be effective for inhibiting COX-1, COX-2, and iNOS. These results suggested that anthocyanin and ternatin flavonoids may potentially be developed as anti-inflammatory drug candidate for the treatment of inflammatory-related diseases.
Photoaging damages skin tissue via activation of matrix metalloproteinases (MMPs) and MMP-3 induc... more Photoaging damages skin tissue via activation of matrix metalloproteinases (MMPs) and MMP-3 induces activation of MMP-1 and 9. Tempeh is made from fermented soybean in Indonesia and has been known for its high protein content. This study was aimed to investigate the effectiveness of peptide extracts from tempeh against photoaging at gene level in UVB-treated mice. Peptide was extracted from soybean and black soybean tempeh, and identified for its amino acid profiling. Photoaging model was made by gradient UVB exposure to seven groups of mice for 4 weeks. Mice were supplemented with 100 and 300 mg/kg body weight dosages of peptide extracts from soybean and black soybean tempeh. Dorsal skin tissue was collected for histology staining and quantitative polymerase chain reaction (qPCR). This present study found that both tempeh peptide extracts were rich in glutamic acid and branched-chain amino acids. Histology staining showed that tempeh protected mice skin from photoaging. At 100 and 300 mg/kg, both tempeh peptide extracts significantly attenuated MMP-3gene expression that lead to the decrease level of MMP-1 and MMP-9 gene expression. These results suggest that supplementation of both tempeh peptide extracts may possess anti-photoaging effect via altering MMP-3 gene expression in UVB-treated mice.
Halitosis is associated with the excessive production of acid and volatile sulphur compounds (VSC... more Halitosis is associated with the excessive production of acid and volatile sulphur compounds (VSCs), as well as accumulation of biofilm plaque by oral bacteria in mouth cavity. Zanthoxylum acanthopodium, locally known as andaliman or lemon pepper, is an Indonesian endemic spice with high content of essential oils, including carveol that has been reported for its potential antimicrobial and anti-inflammatory activities. We determined whether extract of Z. acanthopodium essential oil (ZAEO) exerted anti-halitosis effect on reducing biofilm plaque, acid production, and VSCs by using Actinomyces viscosus model in vitro. ZAEO was extracted in n-hexane followed by evaporation. ZAEO at various doses (20-100 μg/ml) was tested and quantified for its antibiofilm, acid production, and total VSCs production toward A. viscosus oral bacteria in vitro by conducting biofilm assays for preventing and eradicating effects, pH-stat analysis, and VSCs assay. At lowest dose (20 μg/ml), ZAEO inhibited and removed >50% of A. viscosus biofilms. A. viscosus was able to produce acid rapidly in 20 minutes, resulting in the pH terminal of 5.57, and ZAEO treatment at 40 μg/ml exerted significant inhibition on acid production with the terminal pH of 5.93, respectively. ZAEO at lowest dose also reduced >50% of total VCSs produced by A. viscosus. Our results suggest that ZAEO extract could be applied as a natural ingredient for halitosis treatment.
In vitro antibacterial activity of essential oils from twelve aromatic plants from East Kalimanta... more In vitro antibacterial activity of essential oils from twelve aromatic plants from East Kalimantan, Indonesia. Biodiversitas 20: 2039-2042. In the aim of this works was to investigate the antibacterial activity of twelve essential oils against Streptococcus mutans and Streptococcus sobrinus, oral pathogen causing dental caries. The essential oils were collected by a steam distillation method. Refractive index was measured by using a refractometer. The antibacterial activity of essential oils was determined using an agar well diffusion method. The yields of essential oils varied with the plant species. The steam distillation yielded clear to yellowish essential oils. Refractive indexes of oil were found to be i n the range of 1.344 to 1.500. Syzigium sp. appeared to be more abundant in oil (1.54%) than the other plant species, while C. odorata was poorer (0.04%). All of the essential oils tested showed a varied level of inhibition zone (0-53.65 mm) against S. mutans and S. sobrinus. The oil from leaves of Cymbopogon citratus had the highest activity against S. sobrinus and S. mutans (53.15 and 52.85 mm, respectively). On the other hand, the Magnolia x alba oil showed the lowest activity against S. mutans and S. sobrinus (10.50 and 11.65 mm, respectively). The research results demonstrated that the essential oil in this study has the potency for development of dental health products for preventing and treating oral infections.
Curcuminoid cider fermented from Curcuma xanthorrhiza curcuminoids attenuates gene expression rel... more Curcuminoid cider fermented from Curcuma xanthorrhiza curcuminoids attenuates gene expression related to obesity-induced inflammation in hypercholesterolaemic rats Abstract Obesity is an important risk factor in the development of atherosclerosis, and often associated with hypercholesterolemia. Curcuminoid cider, a fermented beverage made from curcuminoid fraction isolated from Curcuma xanthorrhiza, has been reported to have high contents in organic acids and curcuminoids. Herein, we investigated whether curcuminoid cider affected the expression of genes related to obesity-induced inflammation (peroxisome proliferator-activated receptors γ (PPARγ), CCAT/enhancer binding protein α (C/EBPα), fatty acid-binding protein 4 (FABP4)) and genes generating pro-inflammatory cytokines (interleukin (IL) 1β (IL1β), tumour necrosis factor α (TNFα) and chemokine) in hypercholesterolaemic rats by conducting quantitative real time-PCR (qRT-PCR). Twenty-four male Sprague-Dawley rats were divided into six groups: normal group diet, high cholesterol diet (HCD), HCD+ 1% v/v curcuminoid cider, HCD + 2% v/v curcuminoid cider, HCD + 100 mg/kg bw curcuminoid fraction, and HCD + 300 mg/kg bw curcuminoid fraction for four weeks. Rats' lung and liver were collected for total RNA extraction, followed by quantitative analysis to determine gene expression related to obesity and pro-inflammatory cytokines. Results showed that curcuminoid cider at 1 and 2% v/v significantly reduced >80% gene expression of obesity-induced inflammation, including PPARγ, C/EBPα, FABP4, IL1β, TNFα and chemokine in hypercholesterolaemic rats. Meanwhile, curcuminoid fraction at 100 and 300 mg/kg was effective to attenuate up to 80% PPARγ gene expression correlated to obesity. These results indicate that curcuminoid cider may exert antiobesity potential at molecular level through suppressing various genes related to obesity and pro-inflammatory cytokines in hypercholesterolaemic rats. It may be used as a potential candidate in functional beverage for management of obesity-induced inflammation.
Polyunsaturated fatty acids (PUFAs) are natural substances that are considered for skin protectio... more Polyunsaturated fatty acids (PUFAs) are natural substances that are considered for skin protection caused by UVB exposure. The increased levels of photoaging and inflammation markers, i.e. matrix metalloproteinases (MMPs), cyclooxygenase (COX)-2, and inducible nitric oxide synthase (iNOS) are correlated with the signs of photoaging and inflammation in the skin. Tempeh, one of the Indonesian fermented foods is a promising source of PUFAs for food applications including nutraceuticals and cosmeceuticals. In this study, PUFA-rich oil was extracted from white and black soybean tempeh and tested on its efficacy on down-regulating several genes related to photoaging and inflammation in UVB-irradiated premature skin aging in mice model by conducting hematoxylin and eosin (H&E) staining and quantitative Real Time-PCR (qRT-PCR) assays. Tempeh oil was extracted in methanol and chloroform using Bligh and Dyer method, and PUFA constituents in tempeh oil were determined by pyrolysis gas chromatography-mass spectrometry (py/GC-MS) analysis. Mice skin dorsal was irradiated with UVB gradually for 4 weeks to induce premature photoaging. Mice were grouped into negative control; positive control; black soybean tempeh oil at 100 and 300 mg/kg; white soybean tempeh oil at 100 and 300 mg/kg; and omega 3 standard. Mice were orally supplemented with tempeh oil for 4 weeks along with UVB irradiation. The py/GC-MS profiling revealed that both tempeh oils contained major linoleic acid PUFA with 52.3% in white tempeh oil and 85.69% in black tempeh oil, respectively. H&E staining demonstrated that UVB irradiation caused premature skin photoaging in mice, indicating by necrosis, epidermal atrophy, and tissue damage. Orally tempeh oil treatment improved tissue damage and increased the proliferation of hair follicle in the skin. Tempeh oil was found to be effectively inhibited the genes expression of MMP-1, MMP-3, MMP-9, COX-2 and iNOS in UVB-irradiated mice skin. Among these genes, tempeh oil from black and white soybean tempeh exerted a significant effect on down-regulating the expression of MMP-9 gene in UVB-irradiated mice skin. In conclusion, tempeh oil may offer the promising photoprotective ingredient in application of nutraceuticals and cosmeceuticals for protecting skin photoaging.
Halitosis is caused by oral bacteria including Streptococcus sanguinis in mouth producing volatil... more Halitosis is caused by oral bacteria including Streptococcus sanguinis in mouth producing volatile sulphur compounds (VSCs), such as hydrogen sulfide, ethyl mercaptan, and methyl mercaptan which have pungent odor. Bacteria producing sulphur compounds produce oral biofilms as the accumulation of caries promotion. Caries is caused by the acid produced by oral bacteria that lead to tooth demineralization in low pH condition. Clove bud (Syzygium aromaticum), known as endogenous spice in Indonesia, has been traditionally used for centuries for treatment of periondal diseases. In this study, we extracted essential oil from Syzygium aromaticum (SAEO), identifed for its major essential oils by pyrolysis gas chromatograpy mass spectrometry (py-GC/MS), determined its antihalitosis efficacy on preventing and eradicating S. sanguinis oral biofilms, inhibiting VSCs and acid production in vitro. Chromatogram profile showed that SAEO contained major eugenol (22.10%) and aceteugenol (13.31%). For antibiofilm effect toward oral bacteria S. sanguinis, SAEO at 40 and 60 μg/mL effectively prevented biofilm formation up to 60% and removed the existed biofilms up to 50%. SAEO at 60 μg/mL also demonstrated a significant inhibition on VSCs production (up to 58%) and acid produced by S. sanguinis by increasing the terminal pH from 5.66 to 6.30. These data suggest that SAEO could be applied for a promising candidate for developing oral care functional products for management of halitosis.
Inflammation plays an important role in the development of type 2 diabetes mellitus (T2DM), inclu... more Inflammation plays an important role in the development of type 2 diabetes mellitus (T2DM), including obesity-related insulin resistance. Biomarkers of inflammation, such as C-reactive protein (CRP), tumor necrosis factor alpha (TNF-α), interleukin-6, and monocyte chemoattractant protein-1 (MCP-1) are present at increased concentrations in insulin resistant and obese individuals. In this study, we investigated the antidiabetic activity of Zanthoxylum acanthopodium essential oil (ZAEO) on attenuation of proteins related to diabetes in hepatocytes in vitro. ZAEO was extracted from Z. acanthopodium fruits using hexane and identified by using pyrolysis gas chromatography-mass spectrometry (py-GC/MS). ZAEO was tested for its cytotoxicity against human Chang liver cells as an in vitro cultured hepatocytes model. Effect of ZAEO (1-50 μg/mL) on protein expression related to T2DM, including CRP, TNF-α, and MCP-1 was quantified by enzyme-linked immunosorbent assay (ELISA) assay. Chromatogram profile demonstrated that ZAEO mainly consisted of carveol (~47.7%). MTT profile showed that safest dose of ZAEO against hepatocytes was reached up to 10 μg/mL. ELISA data showed that among three proteins, ZAEO exerted significant inhibitory effect against MCP-1 protein expression secreted by hepatocytes. At lowest dose (1 μg/mL), ZAEO attenuated MCP-1 expression up to 30%, respectively. These findings suggest that ZAEO may down-regulate inflammation related to T2DM through inhibiting MCP-1 expression that leads to the increase of insulin-stimulated glucose uptake.
Breastfeeding mothers need more nutrients compared to mothers who are not breastfed to maintain q... more Breastfeeding mothers need more nutrients compared to mothers who are not breastfed to maintain quality and quantity of human milk they produce. Black soybean and black rice contain carbohydrate, protein, fat, and other nutrients that needed by breastfeeding mothers. The purpose of this study was to develop a snack bar for breastfeeding mothers. Snack bars were made from three ratio variations of black soybean flour: black rice flour (30:70, 50:50, and 70:30) with baking temperature 150 0 C for 20 minutes. They were then subjected to sensory analysis (taste, flavor, color, texture, aftertaste, and overall); physical analysis (hardness, color); and chemical analysis (nutri-tional analysis, genistein, and daidzein). The results showed that snack bar with ratio 30:70 (black soybean: black rice) was preferred by panelists. Its level of hardness was 6.10±0.01 N. Its color was showed in color index L* = 25.04±1.71, a* = 6.05±0.70, b* = 3.89 ± 0.71, chroma = 6.66±0.25, and hue = 54.70±2.40. One serving (30 gram/serving) of this snack bar contained 16.28 grams of carbohydrate; 3.10 grams of protein; and 6.05 grams of fat. It could suffice 4.98 % of total energy based on Recommended Dietary Allowance of Indonesian breastfeeding mother per day. Meanwhile, the content of daidzein and genistein were 0.09 mg/serving and 1.35 mg/serving, respectively. In conclusion, snack bar from black soybean and black rice can be used as a complementary food for breastfeeding mothers due to its nutrient content and acceptability.
Soybean milk is a derivative product from soybean that has been known for its functionality effec... more Soybean milk is a derivative product from soybean that has been known for its functionality effects on human. However further development is needed to improve its organoleptic quality. Fermentation is known for reducing beany flavor on soybean milk, on the other hand diminishing its shelf life. In this study, the effects of soybean milk fermentation using Lactobacillus bulgaricus and Streptococcus thermophilus, and post-fermentation sterilization on quality of soybean milk were investigated. Three soybean milk products, i.e. soybean milk (SM), fermented soybean milk (FSM), and sterilized fermented soybean milk (SFSM) were tested for their physico-chemical properties, proximate analysis, amino acid composition, vitamin B 12 level, antioxidant activity, and organoleptic test. Our results showed that fermentation in FSM effectively reduced pH and total soluble solids, increased viscosity, changed amino acid composition, and increased antioxidant activity compared to those of SM. Interestingly, FSM had higher contents of methionine and cysteine than SM. Meanwhile, post-fermentation sterilization in SFSM changed the texture of fermented soybean milk, degraded heat-sensitive compounds, and reducing the antioxidant activity. However, organoleptic profiles for both FSM and SFSM showed a slight decrease in consumer acceptance compared to that of SM. In summary, fermention and sterilization treatments may contribute to the increased quality of soybean milk.
Periodontitis, an inflammatory disease of the gingival tissue, triggered by microbial-derived ele... more Periodontitis, an inflammatory disease of the gingival tissue, triggered by microbial-derived elements, such as lipopolysaccharide (LPS), collapses the periodontal tissues and resorbs the alveolar bone. This study evaluated the inhibitory effects of standardized Boesenbergia pandurata extract (BPE) and panduratin A (PAN) on periodontitis-induced inflammation and alveolar bone loss. Sprague-Dawley rats with LPS-induced periodontitis were orally administered BPE (50 and 200 mg/kg/ day) and PAN (20 mg/kg/day) for 8 days. Histological analysis revealed that BPE-and PAN-administered groups showed decreased cell infiltration and alveolar bone resorption. Furthermore, the BPE and PAN significantly alleviated the mRNA and protein expression levels of nuclear factor kappa B (NF-jB), interleukin-1b, matrix metalloproteinase (MMP)-2, and MMP-8. BPE and PAN also inhibited the expression of nuclear factor of activated T cells, cytoplasmic 1, c-Fos, and ostoclastogenesis-related enzymes, including cathepsin K and tartrate-resistant acid phosphatase (ALP). BPE and PAN not only upregulated the osteoblastogenesis-associated markers, such as collagen type I (COL1A1) and ALP, but also increased the ratio of osteopro-tegerin to receptor activator of NF-jB ligand. Collectively, BPE and PAN efficiently prevent destruction of periodontal tissues and stimulating the loss of alveolar bone tissues, strongly indicative of their potential as natural antiperiodontitis agents.
Fermented foods have been explored for their potentials as main sources of fibrinolytic enzymes. ... more Fermented foods have been explored for their potentials as main sources of fibrinolytic enzymes. In this study, we did screening for fibrinolytic enzymes-producing bacteria isolated from several Indonesian fermented foods, including black tauco, salted tauco, dadih, cassava tape, brem, salted vegetable, fermented fish, terasi, tempe, oncom, fermented fish, tahu, and tempoyak. Fibrinolytic activity was directly determined by fibrin zymogram and fibrin-plating assays. Crude enzyme from selected isolate producing optimal fibrinolytic activity was produced, purified, and further characterized for its biochemical properties (optimum pH, optimal temperature, protease inhibitor, metal ion, and substrate specificity). Crude enzyme was purified using ammonium sulphate fractionation with 65% saturation, dyalisis using nitrocellulose acetate membrane (cut off 10 kDa), concentration with polyethylene glycol, followed by fractionation and purification using fast protein liquid chromatography with DEAE-Sepharose column. Our results demonstrated that a fibrinolytic enzyme (F7 eluate) was successfully isolated, purified, and characterized from the culture filtrate of TH-5 strain isolated from the black tauco. Spesific activity of F7 eluate was 15.421 U/mg with 45.6 purification fold compared to its crude enzyme. The optimum pH and temperature for enzyme activity of F7 eluate were 7 and 50 o C, respectively. Enzyme activity of F7 eluate was totally inhibited by addition of phenylmethylsolfonylfluoride (PMSF) and N-α-tosyl-L-lysine chloromethyl ketone (TLCK) inhibitors at final concentration of 1 mM, indicating that enzyme was grouped in serine protease family. Metal ions (K + , Na + , Mg 2+ , Mn 2+ , and Cu 2+) at total concentration of 5 mM strongly reduced enzyme activity with residual activity less than 30%, indicating that enzyme catalytic of F7 eluate was significantly inhibited by the addition of the ions. Fibrin zymogram profile of F7 eluate resulted in single fibrinolytic band with estimated molecular weight of 36.2 kDa, and the enzyme had a specific characteristic to degrade both fibrin and fibrinogen substrates.
Acne vulgaris has been a skin disease that is caused by excessive oil on the skin that gives an o... more Acne vulgaris has been a skin disease that is caused by excessive oil on the skin that gives an optimal environment for acne-causing skin microbes, including Propionibacterium acnes. Sugar palm fruit (Arenga pinnata) has the potential for acne treatment due to its bioactive compounds that have been reported for exerting anti-inflammation and antioxidant activities. This research was focused on investigation of antibacterial and antibiofilm activities of sugar palm fruit extract (SPFE) for management of acne caused by P. acnes. Sugar palm fruit was extracted in methanol to produce SPFE and the chemical compounds of SPFE were identified by using pyrolysis gas chromatography-mass spectrometry (py-GC/MS). Bioefficacy of SPFE as antibacterial and antibiofilm agents was tested on inhibiting P. acnes growth, eradicating the existed P. acnes biofilms, and preventing P. acnes biofilm formation in vitro. Chromatographic profiling by py-GC/MS showed that SPFE consisted of major compounds, including levoglucosan and methyl-β-D-glucoside as pyran group. SPFE at 16 µg/mL effectively inhibited ~40% of P. acnes growth. For antibiofilm effect, SPFE was more effective for eradication effect on existed P. acnes biofilm compared to that of preventive effect on P. acnes biofilm formation. At 100 µg/mL, SPFE removed up to 50% of the existed P. acnes biofilms. Thus, SPFE may offer alternative candidate to treat acne vulgaris caused by P. acnes by inhibiting its growth and removing its existed biofilms.
Many health related problems such as cardiovascular diseases are associated with the formation of... more Many health related problems such as cardiovascular diseases are associated with the formation of excessive clot in the blood (thrombus). Approaches in cardiovascular disease treatment are preventing the formation or removing the thrombus. The present thrombolytic agents can be classified as plasminogen activators, fibrinolytic enzyme which directly degrades fibrinogen or fibrin and heparin type which act as thrombin inhibitor. Recently, microbial fibrinolytic enzymes of food origin receive more attention that leads to escalating efforts to explore traditional fermented foods as the natural sources. We have successfully isolated microorganism from Indonesian fermented soybean tofu dregs " Oncom " that secretes fibrinolytic enzyme. The microorganism identified as Stenotrophomonas sp. is unique because most of the reported fibrinolytic microorganism belongs to Bacillus sp. This isolate was found to produce extracellular fibrinolytic enzyme which could degrade fibrinogen and fibrin directly as determined by fibrinogen zymography and fibrin plate methods. More importantly, the 30-kD purified enzymes was found to demonstrate not only fibrin and fibrinogen degradation capabilities, but also acted as thrombin inhibitor as determined using specific substrates for thrombin. This is the first report of a fibrinolytic enzyme that demonstrates additional synergistic activities. This finding accentuates the importance of further development of the enzyme into a powerful agent to treat the thrombus-related disease effectively.
Obesity is a prevalent metabolic disorder that is induced by inflammatory responses that lead to ... more Obesity is a prevalent metabolic disorder that is induced by inflammatory responses that lead to the increased levels of pro-inflammatory mediators, including interleukin-6 (IL-6) and leptin. Pithecellobium jiringa or jengkol, is a functional tropical plant in Southeast Asia regions including Indonesia that has been for culinary and traditional medicines. This research was aimed to identify chemical constituents of protein fractions from P. jiringa peel and seed by using pyrolysis gas chromatography mass spectrometry (py-GC/MS), and their inhibitory effects at various concentrations (5-100 µg/ml) on IL-6 and leptin expression in mouse 3T3-L1 preadipocytes by conducting the enzyme-linked immunosorbent assay (ELISA). Both protein fractions from P. jiringa peel and seed consisted of amino acid derivatives, including alanine ethyl ester (peel part), dimehylamine, piperidine, aziridine, and phenyl acetyl glutamine (seed part). Protein fractions from P. jiringa peel at 25 µg/ml decreased >50% leptin expression in 3T3-L1 preadipocytes compared to that of seed protein fractions. Meanwhile, seed protein fraction dose-dependently inhibited IL-6 expression. At 10 µg/ml, seed protein fraction reduced up to 50% IL-6 production, respectively. Our results suggest that protein fractions from both P. jiringa peel and seed may have potential anti-obesity effects on 3T3-L1 preadipocytes via attenuating IL-6 and leptin expression. P. jiringa protein fractions can be used as alternative food supplements for reducing the risk of obesity.
In previous study, 141 isolates of marine bacteria were successfully isolated from Indonesian coa... more In previous study, 141 isolates of marine bacteria were successfully isolated from Indonesian coastal sediment. This study was aimed to investigate whether crude extracts of these selected isolates exerted potential antimicrobial and antioxidant activities in vitro. Crude extract was prepared by solvent extraction using 1-butanol. Antimicrobial assay of crude extracts (1-500 µg/mL) against Bacillus cereus and Staphylococcus aureus was determined by using Minimum Inhibitory Concentration (MIC). Antioxidant assay of crude extracts (5-500 µg/mL) was done by employing 3 assays, including 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferrous chelating activity (FCA), and anti-lipid peroxidation. Our results demonstrated that crude extracts of isolates exerted moderate antimicrobial activities against growth of B. cereus and S. aureus with MIC values > 500 µg/mL. Some isolates showed potential antioxidant activity in a independent-dose manner. Among 3 antioxidant systems, crude extracts of isolates at 500 µg/mL exerted antioxidant activity < 45% using DPPH assay, < 35% using FCA assay, and < 75% using anti-lipid peroxidation assay. Based on molecular identification using 16S rRNA gene sequence, these isolates with antimicrobial and antioxidant properties were identified as Psychrobacter alimentarius, Bacillus sp. and B. niabensis. These results suggest that crude extracts of P. alimentarius may potentially be an alternative antimicrobial and antioxidant agents.
Skin is an important barrier that protects the body from damage due to its direct contact with th... more Skin is an important barrier that protects the body from damage due to its direct contact with the outside environment. Skin has epidermal units that are responsible for melanin production and distribution, a process called melanogenesis that is regulated by various enzymes, including tyrosinase. Due to its central role in melanogenesis, tyrosinase is a key target for screening and discovery of new inhibitors. The aim of this study was to investigate the efficacy of purified protein fractions from soybean (Glycine max) on inhibiting tyrosinase in vitro. Protein was extracted by using isoprecipitation method, followed by protein purification using Fast Protein Liquid Chromatography (FPLC) with DEAE anion exchange system and gradient NaCl elution. Among 9 purified protein fractions, there were 2 protein fractions (10 µg/mL) with molecular weights of 42 and 46 kDa exerted significant inhibition up to 95% on tyrosinase activity in vitro. In conclusion, purified protein fraction from soybean demonstrated potential tyrosinase inhibitory effect, indicating their potential use on cosmeceutical application.
Endometriosis prevalence has been known to be quite high among women of reproductive age and with... more Endometriosis prevalence has been known to be quite high among women of reproductive age and with pelvic pain and/or infertility. The reason is that the estrogen level in the eutopic endometrium of women with endometriosis is higher than in normal endometrium which may possibly be caused by the lack of interaction between progesterone and progesterone receptor (PR). Dexa Laboratories of Biomolecular Sciences (DLBS) has developed DLBS1442, a bioactive fraction from Phaleria macrocarpa (Scheff) Boerl fruit, which has been found to be potential to treat symptoms of primary dysmenorrhea and alleviate endometriosis. Therefore, the identification of DLBS1442 active compounds which act as a PR agonist was necessary. Identification was performed using metabolomics study which resulted in 14 compounds. Crystal structure of the PR with asoprisnil as the reference was obtained from PDB (4A2J). Virtual screening validation process was performed using Protein-Ligand ANT System (PLANTS) and Python-based Protein-Ligand Interaction Fingerprinting (PyPLIF). According to the virtual screening protocol validation, the highest Enrichment Factor (EF) 1% value was obtained with hydrogen interaction with GLN725 and ARG766 residue. Virtual screening of the DLBS1442 metabolomics study showed that only glyceryl pentacosanoate exhibited a lower Chem Piecewise Linear Potential (ChemPLP) than the cutoff. This compound might have a role as a PR agonist which supported the previous findings of DLBS1442 to alleviate endometriosis. However, this finding requires further in vitro and/or in vivo study to ensure the agonist activity of glyceryl pentacosanoate as a DLBS1442 active compound.
Inflammation is characterized by an increase in vascular permeability leading to edema formation.... more Inflammation is characterized by an increase in vascular permeability leading to edema formation. Phosphoinositide 3-kinase (PI3K) signaling pathway plays the main role to regulate several key events in the inflammatory response to damage and infection. Our previous in silico study showed that anthocyanin and ternatin flavonoids may offer anti-inflammatory potential via molecular docking model. Here, we explored whether Clitoria ternatea anthocyanin extract (CTA) had protective activity against inflammation via regulating gene expression related to PI3Ks in rat edema. CTA was extracted in methanol and acetone using maceration and lyophilization. Edema model was done by injecting carrageenan into rat paw. Oral supplementation of CTA (100, 250, and 500 mg/kg BW) and ibuprofen standard (15 mg/kg BW) was given to rats daily for 7 days. After sample treatment, the right hind paw of the rat was injected with carrageenan to induce edema condition. The ability of CTA to suppress the inflammation in paw edema was measured by quantifying the increased percentage of rat paw volume at 1, 3, and 5 hrs after injection. Rat paw tissue was collected, and gene expression related to PI3Ks, i.e. Akt1, PKB, Ilk, Pdk2, Pik3Ca, and p53 was determined using quantitative PCR (qPCR). Carrageenan showed a time-dependent increase in volume of rat paw starting from injection until the next 5 hrs. It reached maximum volume increase of rat paw at 3 hrs after injection. CTA treatment had a dose-dependent pattern in reducing the edema volume in carrageenan-induced paw edema. At 500 mg/kg BW, CTA significantly suppressed the inflammation in paw edema compared to that of positive control, indicating by the low percentage of paw edema volume at 1, 3, and 5 hrs after carrageenan injection. At gene level, qPCR data showed that CTA significantly down-regulated the expression of genes involved in PI3Ks, such as Akt1, PKB, Ilk, Pdk2, Pik3Ca, and p53 in paw tissues. Thus, CTA may have a protective effect against edema via suppressing genes of PI3Ks.
Butterfly pea (Clitoria ternatea) is an indigenous plant from Moluccas in Indonesia. Due to its p... more Butterfly pea (Clitoria ternatea) is an indigenous plant from Moluccas in Indonesia. Due to its pharmacological properties, the flower of this plant is also used as a medicine in Ayurvedic practices in India. The present work was aimed to determine the antidiabetic activity of C. ternatea protein extract (PCT) via the inhibition of α-amylase activity in vitro, and the modulation of genes related to diabetes in diabetic mice in vivo. C. ternatea flower protein extract was prepared using isoprecipitation, and its efficacy as an antidiabetic candidate was tested in vitro through α-amylase inhibition assay, and in vivo through the measurement of blood glucose level and diabetes-related genes expression, including those that encode for peroxisome prolif-erator-activated receptors gamma (PPARγ), glucose transporter 2 (Glut2), transcription factor 7-like 2 (Tcf7l2), calpain-10 (Capn10), and monocyte chemoattractant protein 1 (MCP1) in alloxan-induced diabetic mice. PCT showed α-amylase inhibition in a dose-dependent manner. The highest α-amylase inhibition (20.63%) was observed at 1 mg/mL of PCT. The in vivo study showed that PCT decreased the blood glucose level in alloxan-induced diabetic mice. PCT also altered the expression of diabetes-related genes. In adipose and skeletal muscle tissues, PCT upregulated the gene expression of PPARγ and Tcf7l2, meanwhile MCP1 expression was downregulated. In the pancreas, PCT upregulated the gene expression of Glut2, Capn10, and Tcf7l2. These results indicate that PCT is a potential antidiabetic agent for the treatment of type 2 diabetes.
Inflammatory markers such as cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), myel... more Inflammatory markers such as cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), myeloperoxidase (MPO), and prostaglandin (PEG) are widely known as major targets in discovering natural anti-inflammatory drugs for the treatment of inflammation-related diseases. Non-steroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen and aspirin are mostly used at present, however, some NSAIDS have been reported to cause gastrointestinal side effect due to ligand-protein interaction. Molecular docking is a promising tool to study such modes of interaction. In this study, we evaluated the potential use of anthocyanin and ternatin flavonoids as natural anti-inflammatory agents for treatment of inflammatory-related diseases using in silico molecular docking assay. Automated docking study using Protein-Ligand ANT System (PLANTS) and AutoDock Vina was performed with various ligand molecules, including ibuprofen, anthocyanin, and ternatin against the protein crystal structures of COX-1, COX-2, iNOS, and MPO. The in silico data demonstrated that ibuprofen bound effectively to the active site of COX-1 and MPO with minimum binding energy, yet the compound required more energy to bind the active site of COX-2. Ternatin flavonoid was bound to COX-2 and iNOS with minimum binding energy. In terms of binding energy, anthocyanin flavonoid was found to be effective for inhibiting COX-1, COX-2, and iNOS. These results suggested that anthocyanin and ternatin flavonoids may potentially be developed as anti-inflammatory drug candidate for the treatment of inflammatory-related diseases.
Photoaging damages skin tissue via activation of matrix metalloproteinases (MMPs) and MMP-3 induc... more Photoaging damages skin tissue via activation of matrix metalloproteinases (MMPs) and MMP-3 induces activation of MMP-1 and 9. Tempeh is made from fermented soybean in Indonesia and has been known for its high protein content. This study was aimed to investigate the effectiveness of peptide extracts from tempeh against photoaging at gene level in UVB-treated mice. Peptide was extracted from soybean and black soybean tempeh, and identified for its amino acid profiling. Photoaging model was made by gradient UVB exposure to seven groups of mice for 4 weeks. Mice were supplemented with 100 and 300 mg/kg body weight dosages of peptide extracts from soybean and black soybean tempeh. Dorsal skin tissue was collected for histology staining and quantitative polymerase chain reaction (qPCR). This present study found that both tempeh peptide extracts were rich in glutamic acid and branched-chain amino acids. Histology staining showed that tempeh protected mice skin from photoaging. At 100 and 300 mg/kg, both tempeh peptide extracts significantly attenuated MMP-3gene expression that lead to the decrease level of MMP-1 and MMP-9 gene expression. These results suggest that supplementation of both tempeh peptide extracts may possess anti-photoaging effect via altering MMP-3 gene expression in UVB-treated mice.
Halitosis is associated with the excessive production of acid and volatile sulphur compounds (VSC... more Halitosis is associated with the excessive production of acid and volatile sulphur compounds (VSCs), as well as accumulation of biofilm plaque by oral bacteria in mouth cavity. Zanthoxylum acanthopodium, locally known as andaliman or lemon pepper, is an Indonesian endemic spice with high content of essential oils, including carveol that has been reported for its potential antimicrobial and anti-inflammatory activities. We determined whether extract of Z. acanthopodium essential oil (ZAEO) exerted anti-halitosis effect on reducing biofilm plaque, acid production, and VSCs by using Actinomyces viscosus model in vitro. ZAEO was extracted in n-hexane followed by evaporation. ZAEO at various doses (20-100 μg/ml) was tested and quantified for its antibiofilm, acid production, and total VSCs production toward A. viscosus oral bacteria in vitro by conducting biofilm assays for preventing and eradicating effects, pH-stat analysis, and VSCs assay. At lowest dose (20 μg/ml), ZAEO inhibited and removed >50% of A. viscosus biofilms. A. viscosus was able to produce acid rapidly in 20 minutes, resulting in the pH terminal of 5.57, and ZAEO treatment at 40 μg/ml exerted significant inhibition on acid production with the terminal pH of 5.93, respectively. ZAEO at lowest dose also reduced >50% of total VCSs produced by A. viscosus. Our results suggest that ZAEO extract could be applied as a natural ingredient for halitosis treatment.
In vitro antibacterial activity of essential oils from twelve aromatic plants from East Kalimanta... more In vitro antibacterial activity of essential oils from twelve aromatic plants from East Kalimantan, Indonesia. Biodiversitas 20: 2039-2042. In the aim of this works was to investigate the antibacterial activity of twelve essential oils against Streptococcus mutans and Streptococcus sobrinus, oral pathogen causing dental caries. The essential oils were collected by a steam distillation method. Refractive index was measured by using a refractometer. The antibacterial activity of essential oils was determined using an agar well diffusion method. The yields of essential oils varied with the plant species. The steam distillation yielded clear to yellowish essential oils. Refractive indexes of oil were found to be i n the range of 1.344 to 1.500. Syzigium sp. appeared to be more abundant in oil (1.54%) than the other plant species, while C. odorata was poorer (0.04%). All of the essential oils tested showed a varied level of inhibition zone (0-53.65 mm) against S. mutans and S. sobrinus. The oil from leaves of Cymbopogon citratus had the highest activity against S. sobrinus and S. mutans (53.15 and 52.85 mm, respectively). On the other hand, the Magnolia x alba oil showed the lowest activity against S. mutans and S. sobrinus (10.50 and 11.65 mm, respectively). The research results demonstrated that the essential oil in this study has the potency for development of dental health products for preventing and treating oral infections.
Curcuminoid cider fermented from Curcuma xanthorrhiza curcuminoids attenuates gene expression rel... more Curcuminoid cider fermented from Curcuma xanthorrhiza curcuminoids attenuates gene expression related to obesity-induced inflammation in hypercholesterolaemic rats Abstract Obesity is an important risk factor in the development of atherosclerosis, and often associated with hypercholesterolemia. Curcuminoid cider, a fermented beverage made from curcuminoid fraction isolated from Curcuma xanthorrhiza, has been reported to have high contents in organic acids and curcuminoids. Herein, we investigated whether curcuminoid cider affected the expression of genes related to obesity-induced inflammation (peroxisome proliferator-activated receptors γ (PPARγ), CCAT/enhancer binding protein α (C/EBPα), fatty acid-binding protein 4 (FABP4)) and genes generating pro-inflammatory cytokines (interleukin (IL) 1β (IL1β), tumour necrosis factor α (TNFα) and chemokine) in hypercholesterolaemic rats by conducting quantitative real time-PCR (qRT-PCR). Twenty-four male Sprague-Dawley rats were divided into six groups: normal group diet, high cholesterol diet (HCD), HCD+ 1% v/v curcuminoid cider, HCD + 2% v/v curcuminoid cider, HCD + 100 mg/kg bw curcuminoid fraction, and HCD + 300 mg/kg bw curcuminoid fraction for four weeks. Rats' lung and liver were collected for total RNA extraction, followed by quantitative analysis to determine gene expression related to obesity and pro-inflammatory cytokines. Results showed that curcuminoid cider at 1 and 2% v/v significantly reduced >80% gene expression of obesity-induced inflammation, including PPARγ, C/EBPα, FABP4, IL1β, TNFα and chemokine in hypercholesterolaemic rats. Meanwhile, curcuminoid fraction at 100 and 300 mg/kg was effective to attenuate up to 80% PPARγ gene expression correlated to obesity. These results indicate that curcuminoid cider may exert antiobesity potential at molecular level through suppressing various genes related to obesity and pro-inflammatory cytokines in hypercholesterolaemic rats. It may be used as a potential candidate in functional beverage for management of obesity-induced inflammation.
Polyunsaturated fatty acids (PUFAs) are natural substances that are considered for skin protectio... more Polyunsaturated fatty acids (PUFAs) are natural substances that are considered for skin protection caused by UVB exposure. The increased levels of photoaging and inflammation markers, i.e. matrix metalloproteinases (MMPs), cyclooxygenase (COX)-2, and inducible nitric oxide synthase (iNOS) are correlated with the signs of photoaging and inflammation in the skin. Tempeh, one of the Indonesian fermented foods is a promising source of PUFAs for food applications including nutraceuticals and cosmeceuticals. In this study, PUFA-rich oil was extracted from white and black soybean tempeh and tested on its efficacy on down-regulating several genes related to photoaging and inflammation in UVB-irradiated premature skin aging in mice model by conducting hematoxylin and eosin (H&E) staining and quantitative Real Time-PCR (qRT-PCR) assays. Tempeh oil was extracted in methanol and chloroform using Bligh and Dyer method, and PUFA constituents in tempeh oil were determined by pyrolysis gas chromatography-mass spectrometry (py/GC-MS) analysis. Mice skin dorsal was irradiated with UVB gradually for 4 weeks to induce premature photoaging. Mice were grouped into negative control; positive control; black soybean tempeh oil at 100 and 300 mg/kg; white soybean tempeh oil at 100 and 300 mg/kg; and omega 3 standard. Mice were orally supplemented with tempeh oil for 4 weeks along with UVB irradiation. The py/GC-MS profiling revealed that both tempeh oils contained major linoleic acid PUFA with 52.3% in white tempeh oil and 85.69% in black tempeh oil, respectively. H&E staining demonstrated that UVB irradiation caused premature skin photoaging in mice, indicating by necrosis, epidermal atrophy, and tissue damage. Orally tempeh oil treatment improved tissue damage and increased the proliferation of hair follicle in the skin. Tempeh oil was found to be effectively inhibited the genes expression of MMP-1, MMP-3, MMP-9, COX-2 and iNOS in UVB-irradiated mice skin. Among these genes, tempeh oil from black and white soybean tempeh exerted a significant effect on down-regulating the expression of MMP-9 gene in UVB-irradiated mice skin. In conclusion, tempeh oil may offer the promising photoprotective ingredient in application of nutraceuticals and cosmeceuticals for protecting skin photoaging.
Halitosis is caused by oral bacteria including Streptococcus sanguinis in mouth producing volatil... more Halitosis is caused by oral bacteria including Streptococcus sanguinis in mouth producing volatile sulphur compounds (VSCs), such as hydrogen sulfide, ethyl mercaptan, and methyl mercaptan which have pungent odor. Bacteria producing sulphur compounds produce oral biofilms as the accumulation of caries promotion. Caries is caused by the acid produced by oral bacteria that lead to tooth demineralization in low pH condition. Clove bud (Syzygium aromaticum), known as endogenous spice in Indonesia, has been traditionally used for centuries for treatment of periondal diseases. In this study, we extracted essential oil from Syzygium aromaticum (SAEO), identifed for its major essential oils by pyrolysis gas chromatograpy mass spectrometry (py-GC/MS), determined its antihalitosis efficacy on preventing and eradicating S. sanguinis oral biofilms, inhibiting VSCs and acid production in vitro. Chromatogram profile showed that SAEO contained major eugenol (22.10%) and aceteugenol (13.31%). For antibiofilm effect toward oral bacteria S. sanguinis, SAEO at 40 and 60 μg/mL effectively prevented biofilm formation up to 60% and removed the existed biofilms up to 50%. SAEO at 60 μg/mL also demonstrated a significant inhibition on VSCs production (up to 58%) and acid produced by S. sanguinis by increasing the terminal pH from 5.66 to 6.30. These data suggest that SAEO could be applied for a promising candidate for developing oral care functional products for management of halitosis.
Inflammation plays an important role in the development of type 2 diabetes mellitus (T2DM), inclu... more Inflammation plays an important role in the development of type 2 diabetes mellitus (T2DM), including obesity-related insulin resistance. Biomarkers of inflammation, such as C-reactive protein (CRP), tumor necrosis factor alpha (TNF-α), interleukin-6, and monocyte chemoattractant protein-1 (MCP-1) are present at increased concentrations in insulin resistant and obese individuals. In this study, we investigated the antidiabetic activity of Zanthoxylum acanthopodium essential oil (ZAEO) on attenuation of proteins related to diabetes in hepatocytes in vitro. ZAEO was extracted from Z. acanthopodium fruits using hexane and identified by using pyrolysis gas chromatography-mass spectrometry (py-GC/MS). ZAEO was tested for its cytotoxicity against human Chang liver cells as an in vitro cultured hepatocytes model. Effect of ZAEO (1-50 μg/mL) on protein expression related to T2DM, including CRP, TNF-α, and MCP-1 was quantified by enzyme-linked immunosorbent assay (ELISA) assay. Chromatogram profile demonstrated that ZAEO mainly consisted of carveol (~47.7%). MTT profile showed that safest dose of ZAEO against hepatocytes was reached up to 10 μg/mL. ELISA data showed that among three proteins, ZAEO exerted significant inhibitory effect against MCP-1 protein expression secreted by hepatocytes. At lowest dose (1 μg/mL), ZAEO attenuated MCP-1 expression up to 30%, respectively. These findings suggest that ZAEO may down-regulate inflammation related to T2DM through inhibiting MCP-1 expression that leads to the increase of insulin-stimulated glucose uptake.
Breastfeeding mothers need more nutrients compared to mothers who are not breastfed to maintain q... more Breastfeeding mothers need more nutrients compared to mothers who are not breastfed to maintain quality and quantity of human milk they produce. Black soybean and black rice contain carbohydrate, protein, fat, and other nutrients that needed by breastfeeding mothers. The purpose of this study was to develop a snack bar for breastfeeding mothers. Snack bars were made from three ratio variations of black soybean flour: black rice flour (30:70, 50:50, and 70:30) with baking temperature 150 0 C for 20 minutes. They were then subjected to sensory analysis (taste, flavor, color, texture, aftertaste, and overall); physical analysis (hardness, color); and chemical analysis (nutri-tional analysis, genistein, and daidzein). The results showed that snack bar with ratio 30:70 (black soybean: black rice) was preferred by panelists. Its level of hardness was 6.10±0.01 N. Its color was showed in color index L* = 25.04±1.71, a* = 6.05±0.70, b* = 3.89 ± 0.71, chroma = 6.66±0.25, and hue = 54.70±2.40. One serving (30 gram/serving) of this snack bar contained 16.28 grams of carbohydrate; 3.10 grams of protein; and 6.05 grams of fat. It could suffice 4.98 % of total energy based on Recommended Dietary Allowance of Indonesian breastfeeding mother per day. Meanwhile, the content of daidzein and genistein were 0.09 mg/serving and 1.35 mg/serving, respectively. In conclusion, snack bar from black soybean and black rice can be used as a complementary food for breastfeeding mothers due to its nutrient content and acceptability.
Soybean milk is a derivative product from soybean that has been known for its functionality effec... more Soybean milk is a derivative product from soybean that has been known for its functionality effects on human. However further development is needed to improve its organoleptic quality. Fermentation is known for reducing beany flavor on soybean milk, on the other hand diminishing its shelf life. In this study, the effects of soybean milk fermentation using Lactobacillus bulgaricus and Streptococcus thermophilus, and post-fermentation sterilization on quality of soybean milk were investigated. Three soybean milk products, i.e. soybean milk (SM), fermented soybean milk (FSM), and sterilized fermented soybean milk (SFSM) were tested for their physico-chemical properties, proximate analysis, amino acid composition, vitamin B 12 level, antioxidant activity, and organoleptic test. Our results showed that fermentation in FSM effectively reduced pH and total soluble solids, increased viscosity, changed amino acid composition, and increased antioxidant activity compared to those of SM. Interestingly, FSM had higher contents of methionine and cysteine than SM. Meanwhile, post-fermentation sterilization in SFSM changed the texture of fermented soybean milk, degraded heat-sensitive compounds, and reducing the antioxidant activity. However, organoleptic profiles for both FSM and SFSM showed a slight decrease in consumer acceptance compared to that of SM. In summary, fermention and sterilization treatments may contribute to the increased quality of soybean milk.
Periodontitis, an inflammatory disease of the gingival tissue, triggered by microbial-derived ele... more Periodontitis, an inflammatory disease of the gingival tissue, triggered by microbial-derived elements, such as lipopolysaccharide (LPS), collapses the periodontal tissues and resorbs the alveolar bone. This study evaluated the inhibitory effects of standardized Boesenbergia pandurata extract (BPE) and panduratin A (PAN) on periodontitis-induced inflammation and alveolar bone loss. Sprague-Dawley rats with LPS-induced periodontitis were orally administered BPE (50 and 200 mg/kg/ day) and PAN (20 mg/kg/day) for 8 days. Histological analysis revealed that BPE-and PAN-administered groups showed decreased cell infiltration and alveolar bone resorption. Furthermore, the BPE and PAN significantly alleviated the mRNA and protein expression levels of nuclear factor kappa B (NF-jB), interleukin-1b, matrix metalloproteinase (MMP)-2, and MMP-8. BPE and PAN also inhibited the expression of nuclear factor of activated T cells, cytoplasmic 1, c-Fos, and ostoclastogenesis-related enzymes, including cathepsin K and tartrate-resistant acid phosphatase (ALP). BPE and PAN not only upregulated the osteoblastogenesis-associated markers, such as collagen type I (COL1A1) and ALP, but also increased the ratio of osteopro-tegerin to receptor activator of NF-jB ligand. Collectively, BPE and PAN efficiently prevent destruction of periodontal tissues and stimulating the loss of alveolar bone tissues, strongly indicative of their potential as natural antiperiodontitis agents.
Fermented foods have been explored for their potentials as main sources of fibrinolytic enzymes. ... more Fermented foods have been explored for their potentials as main sources of fibrinolytic enzymes. In this study, we did screening for fibrinolytic enzymes-producing bacteria isolated from several Indonesian fermented foods, including black tauco, salted tauco, dadih, cassava tape, brem, salted vegetable, fermented fish, terasi, tempe, oncom, fermented fish, tahu, and tempoyak. Fibrinolytic activity was directly determined by fibrin zymogram and fibrin-plating assays. Crude enzyme from selected isolate producing optimal fibrinolytic activity was produced, purified, and further characterized for its biochemical properties (optimum pH, optimal temperature, protease inhibitor, metal ion, and substrate specificity). Crude enzyme was purified using ammonium sulphate fractionation with 65% saturation, dyalisis using nitrocellulose acetate membrane (cut off 10 kDa), concentration with polyethylene glycol, followed by fractionation and purification using fast protein liquid chromatography with DEAE-Sepharose column. Our results demonstrated that a fibrinolytic enzyme (F7 eluate) was successfully isolated, purified, and characterized from the culture filtrate of TH-5 strain isolated from the black tauco. Spesific activity of F7 eluate was 15.421 U/mg with 45.6 purification fold compared to its crude enzyme. The optimum pH and temperature for enzyme activity of F7 eluate were 7 and 50 o C, respectively. Enzyme activity of F7 eluate was totally inhibited by addition of phenylmethylsolfonylfluoride (PMSF) and N-α-tosyl-L-lysine chloromethyl ketone (TLCK) inhibitors at final concentration of 1 mM, indicating that enzyme was grouped in serine protease family. Metal ions (K + , Na + , Mg 2+ , Mn 2+ , and Cu 2+) at total concentration of 5 mM strongly reduced enzyme activity with residual activity less than 30%, indicating that enzyme catalytic of F7 eluate was significantly inhibited by the addition of the ions. Fibrin zymogram profile of F7 eluate resulted in single fibrinolytic band with estimated molecular weight of 36.2 kDa, and the enzyme had a specific characteristic to degrade both fibrin and fibrinogen substrates.
Acne vulgaris has been a skin disease that is caused by excessive oil on the skin that gives an o... more Acne vulgaris has been a skin disease that is caused by excessive oil on the skin that gives an optimal environment for acne-causing skin microbes, including Propionibacterium acnes. Sugar palm fruit (Arenga pinnata) has the potential for acne treatment due to its bioactive compounds that have been reported for exerting anti-inflammation and antioxidant activities. This research was focused on investigation of antibacterial and antibiofilm activities of sugar palm fruit extract (SPFE) for management of acne caused by P. acnes. Sugar palm fruit was extracted in methanol to produce SPFE and the chemical compounds of SPFE were identified by using pyrolysis gas chromatography-mass spectrometry (py-GC/MS). Bioefficacy of SPFE as antibacterial and antibiofilm agents was tested on inhibiting P. acnes growth, eradicating the existed P. acnes biofilms, and preventing P. acnes biofilm formation in vitro. Chromatographic profiling by py-GC/MS showed that SPFE consisted of major compounds, including levoglucosan and methyl-β-D-glucoside as pyran group. SPFE at 16 µg/mL effectively inhibited ~40% of P. acnes growth. For antibiofilm effect, SPFE was more effective for eradication effect on existed P. acnes biofilm compared to that of preventive effect on P. acnes biofilm formation. At 100 µg/mL, SPFE removed up to 50% of the existed P. acnes biofilms. Thus, SPFE may offer alternative candidate to treat acne vulgaris caused by P. acnes by inhibiting its growth and removing its existed biofilms.
Many health related problems such as cardiovascular diseases are associated with the formation of... more Many health related problems such as cardiovascular diseases are associated with the formation of excessive clot in the blood (thrombus). Approaches in cardiovascular disease treatment are preventing the formation or removing the thrombus. The present thrombolytic agents can be classified as plasminogen activators, fibrinolytic enzyme which directly degrades fibrinogen or fibrin and heparin type which act as thrombin inhibitor. Recently, microbial fibrinolytic enzymes of food origin receive more attention that leads to escalating efforts to explore traditional fermented foods as the natural sources. We have successfully isolated microorganism from Indonesian fermented soybean tofu dregs " Oncom " that secretes fibrinolytic enzyme. The microorganism identified as Stenotrophomonas sp. is unique because most of the reported fibrinolytic microorganism belongs to Bacillus sp. This isolate was found to produce extracellular fibrinolytic enzyme which could degrade fibrinogen and fibrin directly as determined by fibrinogen zymography and fibrin plate methods. More importantly, the 30-kD purified enzymes was found to demonstrate not only fibrin and fibrinogen degradation capabilities, but also acted as thrombin inhibitor as determined using specific substrates for thrombin. This is the first report of a fibrinolytic enzyme that demonstrates additional synergistic activities. This finding accentuates the importance of further development of the enzyme into a powerful agent to treat the thrombus-related disease effectively.
Obesity is a prevalent metabolic disorder that is induced by inflammatory responses that lead to ... more Obesity is a prevalent metabolic disorder that is induced by inflammatory responses that lead to the increased levels of pro-inflammatory mediators, including interleukin-6 (IL-6) and leptin. Pithecellobium jiringa or jengkol, is a functional tropical plant in Southeast Asia regions including Indonesia that has been for culinary and traditional medicines. This research was aimed to identify chemical constituents of protein fractions from P. jiringa peel and seed by using pyrolysis gas chromatography mass spectrometry (py-GC/MS), and their inhibitory effects at various concentrations (5-100 µg/ml) on IL-6 and leptin expression in mouse 3T3-L1 preadipocytes by conducting the enzyme-linked immunosorbent assay (ELISA). Both protein fractions from P. jiringa peel and seed consisted of amino acid derivatives, including alanine ethyl ester (peel part), dimehylamine, piperidine, aziridine, and phenyl acetyl glutamine (seed part). Protein fractions from P. jiringa peel at 25 µg/ml decreased >50% leptin expression in 3T3-L1 preadipocytes compared to that of seed protein fractions. Meanwhile, seed protein fraction dose-dependently inhibited IL-6 expression. At 10 µg/ml, seed protein fraction reduced up to 50% IL-6 production, respectively. Our results suggest that protein fractions from both P. jiringa peel and seed may have potential anti-obesity effects on 3T3-L1 preadipocytes via attenuating IL-6 and leptin expression. P. jiringa protein fractions can be used as alternative food supplements for reducing the risk of obesity.
In previous study, 141 isolates of marine bacteria were successfully isolated from Indonesian coa... more In previous study, 141 isolates of marine bacteria were successfully isolated from Indonesian coastal sediment. This study was aimed to investigate whether crude extracts of these selected isolates exerted potential antimicrobial and antioxidant activities in vitro. Crude extract was prepared by solvent extraction using 1-butanol. Antimicrobial assay of crude extracts (1-500 µg/mL) against Bacillus cereus and Staphylococcus aureus was determined by using Minimum Inhibitory Concentration (MIC). Antioxidant assay of crude extracts (5-500 µg/mL) was done by employing 3 assays, including 1,1-diphenyl-2-picrylhydrazyl (DPPH), ferrous chelating activity (FCA), and anti-lipid peroxidation. Our results demonstrated that crude extracts of isolates exerted moderate antimicrobial activities against growth of B. cereus and S. aureus with MIC values > 500 µg/mL. Some isolates showed potential antioxidant activity in a independent-dose manner. Among 3 antioxidant systems, crude extracts of isolates at 500 µg/mL exerted antioxidant activity < 45% using DPPH assay, < 35% using FCA assay, and < 75% using anti-lipid peroxidation assay. Based on molecular identification using 16S rRNA gene sequence, these isolates with antimicrobial and antioxidant properties were identified as Psychrobacter alimentarius, Bacillus sp. and B. niabensis. These results suggest that crude extracts of P. alimentarius may potentially be an alternative antimicrobial and antioxidant agents.
Skin is an important barrier that protects the body from damage due to its direct contact with th... more Skin is an important barrier that protects the body from damage due to its direct contact with the outside environment. Skin has epidermal units that are responsible for melanin production and distribution, a process called melanogenesis that is regulated by various enzymes, including tyrosinase. Due to its central role in melanogenesis, tyrosinase is a key target for screening and discovery of new inhibitors. The aim of this study was to investigate the efficacy of purified protein fractions from soybean (Glycine max) on inhibiting tyrosinase in vitro. Protein was extracted by using isoprecipitation method, followed by protein purification using Fast Protein Liquid Chromatography (FPLC) with DEAE anion exchange system and gradient NaCl elution. Among 9 purified protein fractions, there were 2 protein fractions (10 µg/mL) with molecular weights of 42 and 46 kDa exerted significant inhibition up to 95% on tyrosinase activity in vitro. In conclusion, purified protein fraction from soybean demonstrated potential tyrosinase inhibitory effect, indicating their potential use on cosmeceutical application.
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