Factors Influencing Hypoglycemia in Type 2 Diabetes Mellitus Outpatients with State Health Insurance at Regional General Hospitals in Jakarta, Indonesia.

BACKGROUND: Hypoglycemia is an acute episode that can lead to death in patients with diabetes mellitus (DM). This condition is preventable with patient education, and identifying factors influencing their occurrence is essential to creating effective and efficient education. It also leads to prevention and control by re-organizing the service system and diabetes policies. This study aimed to determine factors contributing to hypoglycemic episodes in type 2 DM outpatients covered by the state-provided Jaminan Kesehatan Nasional (JKN) health insurance. METHODS: The study used a cross-sectional design and collected data from five regional general hospitals in Jakarta, Indonesia. The outpatients were sampled consecutively from two hospitals in September-November 2021, one in January-March 2022, and two others in April-June 2023. Interviews produced primary data related to experienced hypoglycemic episodes, and medical records provided secondary data on patients' clinical characteristics and treatments. Binary logistic regression analysis was employed to process the contributing factors statistically. RESULTS: From 501 patients who met the inclusion and exclusion criteria, it was found that the prevalence of hypoglycemia was 53.3%. Factors that significantly increased hypoglycemic risk (p < 0.05) were high HbA1C levels (OR 1.9; 95% CI 1.2-2.9), comorbidities (OR 1.6; 95% CI 1.1-2.4), insulin/sulfonylurea therapy (OR 2; 95% CI 1-4), non-smoking habit (OR 2.2; 95% CI 1.3-3.6) and physically active lifestyle (OR 1.8; 95% CI 1.2-2.6). CONCLUSION: The prevalence of hypoglycemia in type 2 diabetes mellitus (DM) outpatients with the state-provided health insurance Jaminan Kesehatan Nasional (JKN) at general hospitals in Jakarta is high. The diabetes self-management education (DSME) services provided by health professionals for these outpatients must be further improved.

Pharmacognostical and Phytochemical Studies and Biological Activity of Curculigo latifolia Plant Organs for Natural Skin-Whitening Compound Candidate.

Curculigo latifolia (family Amaryllidaceae) is used empirically for medicinal purposes. It is distributed throughout Asian countries, especially Indonesia. This study aimed at standardizing the C. latifolia plant, analyzing its phytochemical profile, and evaluating its pharmacological effects. The powder from each organ (root, stem, and leaves) was standardized organoleptically and microscopically. Samples were extracted by graded maceration using hexane, ethyl acetate, and ethanol. The extracts were determined for total phenolic content (TPC) and total flavonoid content (TFC). Antioxidant (radical scavenging and metal ion reduction) and antityrosinase activities were determined by spectrophotometric methods. Extracts were analysed for phytochemical profiles by LC-ESI-MS. The highest TPC and TFC were found in the ethanolic extract of the root organ (68.63 ± 2.97 mg GAE/g) and the ethyl acetate extract of the stem (14.33 ± 0.71 mg QE/g extract). High antioxidant activities were found in the ethanolic root extract (20.42 ± 0.33 µg/mL) and ethanolic stem extract (45.65 ± 0.77 µg/mL) by DPPH• and NO• assays, respectively. The ion reduction activity (by CUPRAC assay) was most significant in the ethyl acetate stem extract (390.42 ± 14.49 µmol GAEAC/g extract). Ethanolic root extract was the most active in inhibiting tyrosinase (IC50 value of 108.5 µg/mL). The correlation matrix between TPC and antioxidant activities showed a moderate to robust correlation, whereas the TPC and antityrosinase activity showed a robust correlation. The TFC and antioxidant or antityrosinase activities showed a weak to moderate correlation. The LC-ESI-MS data identified major phenols in the active extracts, including methyl 3-hydroxy-4-methoxy-benzoate, quercetin, 4-O-caffeoylquinic acid-1, and curculigoside. Overall, this study suggests that extracts from the C. latifolia plant offer potent antioxidant and antityrosinase activities, allowing them to be used as natural antioxidants and candidates for skin-lightening compounds.

Phytochemical composition, antioxidant, in vitro and in silico studies of active compounds of Curculigo latifolia extracts as promising elastase inhibitor.

Curculigo latifolia is a plant in the Hypoxidaceae family commonly used in herbal medicine. The study objective was to evaluate the antioxidant and anti-elastase properties of C. latifolia extracts in vitro and silico as a candidate for antiaging active ingredients. This study identified secondary metabolites of the hexane (HE), ethyl acetate (EAE), and ethanol extracts (EE) from the root (R), stem (S), and leaf (L) organs by LC-ESI-MS and evaluated in vitro antioxidant and inhibitor elastase activity. An antioxidant evaluation was performed using ABTS, Beta Carotene Bleaching (BCB), and Ferric Reduction Antioxidant Power (FRAP). Evaluation of anti-elastase was carried out using elastase and followed by an in silico study of molecular docking using the target protein elastase (1B0F). Fifteen C. latifolia metabolites were identified in C. latifolia extracts, most of which were phenolic compounds. In antioxidant testing, REE, REAE, SEE, and SEAE extracts showed potent antioxidant activity based on the ABTS, BCB, and FRAP methods. In anti-elastase testing, it was found that SEE, REE, REAE, and RHE extracts gave powerful inhibition of elastase activity (in the ranges of 16.89 to 27.91 µg/mL). The in-silico study demonstrated the potential of the identified metabolites to bind to the target protein 1B0F involved in remodeling the skin aging process. This research concludes that the extracts from C. latifolia have the potential to serve as an active antiaging source.

Efficacy of Using Bacillus subtilis Enzyme as a Caffeine Level Reducer in Cascara Robusta Coffee (Coffea canephora L.).

<b>Background and Objective:</b> An increase in the consumption of robusta coffee resulted in an increase in waste from coffee, one of the coffee wastes, namely coffee bean skins or cascara. Robusta coffee cascara contains 1-1.3 g of caffeine which causes side effects, such as insomnia and seizures etc. So this research aims to reduce the caffeine content in cascara by using <i>Bacillus subtilis</i>. Using optimum conditions and capabilities. <b>Materials and Methods:</b> The experiment was conducted from May to August, 2022 in the Pharmacy Laboratory, Faculty of Mathematics and Natural Sciences, Universitas Pakuan, Indonesia. Before optimizing, cascara was extracted using the ultrasonic assisted extraction (UAE) method, validated by the High-Performance Liquid Chromatography (HPLC) method to determine caffeine content and a paired sample t-test was performed using Statistical Package for the Social Sciences (SPSS). <b>Results:</b> It showed that in validating the HPLC method, the wavelength of caffeine in cascara was 272 nm. The mobile phase was a mixture of methanol-water (adjust orthophosphate). The pH (2.4) (45:55), obtained the optimum decaffeination conditions at the concentration of bacteria <i>Bacillus subtilis</i> 6% and a long incubation time of 24 hrs resulted in a decrease in caffeine content of 51.3843±0.2503%. <b>Conclusion:</b> The results of the paired sample t-test indicate that the concentration of bacteria <i>Bacillus subtilis</i> and incubation time significantly influence caffeine levels.

In vitro studies on the cytotoxicity, elastase, and tyrosinase inhibitory activities of tomato (Solanum lycopersicum Mill.) extract.

Tomatoes (Solanum lycopersicum Mill.), a common vegetable in Indonesia, contain high levels of lycopene, which is good for the body. This research further investigates the activity of polar and nonpolar fractions of tomatoes as elastase and tyrosinase inhibitory, and cytotoxic agents. The extraction procedure used is maceration, fractionation through liquid-liquid fractionation, purification of phytochemical substances is achieved through the application of thin layer chromatography. Elastase and tyrosinase inhibitory activity was analyzed using spectrophotometry and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cytotoxic assay. The result showed that the extract yield was 0.004%. The percentage of polar fraction from the extract was 2.58%, while the nonpolar fraction was 0.69%. The elastase inhibitory activity of polar and nonpolar fractions of tomato extract is 87.21% ± 7.57% and 73.12% ± 7.44%, respectively, The elastase inhibitory activity of polar and nonpolar fractions of tomato extract is 87.21% ± 7.57% and 73.12% ± 7.44%, respectively. The fractions had higher the anti-elastase activity than the positive control quercetin (65.97% ± 3.00%). The tyrosinase inhibitory activity of polar and nonpolar fractions of tomato extract is 23.71% ± 7.91% and 41.16% ± 5.41% (kojic acid as standard is 65.07% ± 0.86%), respectively. The IC50 of the cytotoxic assay to NIH 3T3 mouse embryonic fibroblast cells of the polar and nonpolar fraction of tomato extract is 1820.90 µg/mL and 1643.86 µg/mL, respectively.

Antidiabetic Properties and Toxicological Assessment of Antidesma celebicum Miq: Ethanolic Leaves Extract in Sprague-Dawley Rats.

Antidesma is a genus of plants, and its several species are known to have antidiabetic properties. Leaves of Kayu Tuah (Antidesma celebicum Miq) have been proven to have the best α-glucosidase inhibition ability compared to other species in the Antidesma genus, as evidenced by the in vitro α-glucosidase inhibition test. However, no scientific studies have reported its antidiabetic properties and toxicity in vivo. Therefore, this research managed to verify the antidiabetic features and safety of ethanolic extract of A. celebicum leaves (EEAC) in Sprague-Dawley rats. Male rats (170-280 g) were induced diabetic with streptozotocin (35 mg/kg BW) and fed a high-fat diet comprising 24% fat, whereas control group rats were given a standard diet. Rats were treated with EEAC at 200 and 400 mg/kg BW doses for 28-days and 60 mg/kg BW acarbose for the control group. Determination of antidiabetic properties was done by analyzing lipid profiles as well as fasting blood glucose. After confirming the antidiabetic properties of EEAC, the toxicological assessment was determined using the fixed-dose method. General behavior changes, appearance, signs of toxicity, mortality, and body weight of animals were marked down during the observation period. When the treatment period ended, hematological, biochemical, and histological examinations of liver, kidneys, and heart sections were performed. The results confirmed that EEAC reduced fasting blood glucose levels and stepped forward lipid profiles of rats. Also, all animals survived, and no obvious destructive outcomes were noticeable during the study. As EEAC has promising results toward hyperglycemia and hyperlipidemia and has been proven safe through toxicity tests, it can be concluded that EEAC has good potential to be further developed into antidiabetic drugs.

New robustaflavone from Garcinia latissima Miq. leave and Its antibacterial activity.

Isolation and determination of antibacterial compounds from plants are essential to obtain a new antibacterial as a substitute for conventional resistant antibiotics. This study aims to isolate and identify a new robustaflavone as antibacterial activity from Garcinia latissima Miq. leave. In this study, the isolation process was carried out using column chromatography followed by preparative thin layer chromatography (TLC) based on the TLC profile. The fraction D was tested for anti-bacterial Bacillus subtilis using the TLC bioautography method. The isolates obtained were then identified using 1H-NMR, 13C-NMR, distortionless enhancement by polarization transfer, heteronuclear single quantum coherence, and heteronuclear multiple bond coherence. The Activity assay of the isolate was performed using the microdilution method. A pure compound obtained the result of the separation process with eluent n-hexane: Ethyl acetate (3:2) with Rf 0.6. This spot follows the spot in the contact bioautographic result of fraction D, the spot with Rf 0.6 gives an inhibition zone. After identifying and purifying the isolate were known as Robustaflavone, this compound has activity against B. subtilis with a (minimum inhibitory concentration) value of 2500 ppm. Robustaflavone successfully isolated and identified from G. latissima leave and its antibacterial activity.

The chemical constituents of Sterculia comosa (wall) Roxb woods for arginase inhibitory, antioxidant activity, and molecular docking against SARS CoV-2 protein.

Flavonoids and phenols have an arginase inhibitory and antioxidant activity. The Sterculia genus has phenols and flavonoids content. This study aimed to investigate the arginase inhibitory and antioxidant activity of the chemical constituent of Sterculia comosa (wall) Roxb and also their binding affinities to arginase. The most active extract was methanol extract. This active extract was determined for its arginase inhibitory and antioxidant activity, determined the total phenols and total flavonoids, and identified chemical compound. The methanol extract has IC50 2.787 µg/ml for arginase inhibitory activity and IC50 4,199 µg/ml for DPPH scavenging activity. The total phenols 723.61 mg GAE/gr, total flavonoids content 28.96 mg QE/gr extract. The chemical constituent: KC4.4.6 ((-)-2-(E)-caffeoyl-D-glyceric acid) and KC4.4.5.1 (trans-isoferulic acid) have an arginase inhibitory activity KC4.4.6: 98,03 µg/ml and KC4.4.5.1: 292,58 µg/ml. Antioxidant activity with DPPH methods KC4.4.6: 48,77 µg/ml and KC4.4.5.1: 88,08 µg/ml. Antioxidant by FRAP methods KC4.4.6: 16,4 FeEAC mol/g and KC4.4.5.1: 15,79 FeEAC mol/g. The isolate trans-isoferulic acid predicted has good interaction to arginase. Isolate KC4.4.6. Predicted has good interaction to PLPro of SARS CoV-2 PLpro. However, both isolates did not show good interaction to 3CLPro, nsp12, and Spike protein of SARS CoV-2.

Secondary metabolites from Garcinia daedalanthera Pierre leaves (Clusiaceae).

Two new glycerol esters, (S)-2-hydroxy-3-(octanoyloxy)propyl tetracosanoate (1) and (S)-3-(((S)-11-acetoxy octadecanoyl)oxy)propane-1,2-diyl diacetate (2), and eight known compounds, docosanedioic acid (3), 2,5-dimethylnonadecane (4), lupeol (5), stigmasterol (6), ß-sitosterol (7), heptadecanoic acid (8), hexanedioic acid, 1,6-bis[(2R)-ethylhexyl] ester (9), and 1,3-di-O-[2',2'-di-(p-phenylene)] (10) were isolated from the leaves of Garcinia daedalanthera Pierre, collected from Indonesia. Structural analysis of the isolates was performed using 1 D- and 2 D-NMR, LC- and GC-MS, IR, polarimetry, and UV-visible spectroscopic methods. Cytotoxicity assessments, as well as reactive oxygen species (ROS) analysis of the isolates, were also completed. Lupeol was the only compound found active with an IC50 value of 19.2 µM against HT-29 colon cancer cells. Significant ROS inhibition and induction activity was observed for compounds 4 and 8, respectively.

Antidiabetic activity and acute toxicity of combined extract of Andrographis paniculata, Syzygium cumini, and Caesalpinia sappan.

Andrographis paniculata, Syzygium cumini, and Caesalpinia sappan are used as traditional medicines to treat diabetes mellitus. Therefore, this study aims to examine the antidiabetic effects and the acute toxicity of combined extract (1:1:1) of A. paniculata, S. cumini, and C. sappan (ASCE). The antidiabetic effect was tested using the rats model, induced by a high-fat diet and a double dose of streptozotocin injection of 35 mg/kg BW. Subsequently, diabetic rats in the experimental group were treated with 75 mg/kg BW and 150 mg/kg BW of ASCE, and those in the diabetic control group were treated with metformin 250 mg/kg BW. After seven days of treatment, fasting blood glucose (FBG), pancreatic ß-cells numbers, and lipid profiles were used to analyze the antidiabetic effect. The results showed that the administration of 150 mg/kg BW ASCE significantly reduced FBG (p < 0.01), cholesterol levels (p < 0.05), LDL levels (p < 0.05), but not triglycerides, compared to diabetic control, this effect was comparable to metformin treatment. In addition, the pancreatic ß-cells numbers were likely increased after ASCE treatment in a dose-dependent manner. The oral administration of a single dose of ASCE was safe up to 5000 mg/kg BW and did not result in any significant difference in body weight, relative organ weight, hematological and biochemical parameters compared with the control group. Therefore, it can be concluded that ASCE has a potential antidiabetic effect and can be safely developed as alternative medicine.

Two triterpenoids from Rubus fraxinifolius leaves and their tyrosinase and elastase inhibitory activities.

Numerous therapeutic compounds have been isolated from naturally abundant organic resources, which may offer economical and sustainable sources of compounds with safe and efficacious biological activities. In the cosmetics industry, natural compounds with anti-aging activities are eagerly sought. Thus, we prepared various extracts from Rubus fraxinifolius leaves and used enzyme inhibition assays to isolate compounds with protective effects against skin aging. Two triterpenoids were isolated from Rubus fraxinifolius Poir. leaves. The structures were characterized by spectroscopic analyses (LC-ESI-MS, 1D/2D NMR) and comparison to reported data. Compound 1 and 2 were determined as 2,3-O-ethyleneglycol, 19-hydroxyurs-12-en-23,28-dioic acid and 2,3-O-propanediol,19-hydroxyurs-12-en-28-oic acid. Methanol extract and isolates were assessed for their inhibitory effects on elastase and tyrosinase. Compounds 1 and 2 inhibited elastase with IC50 122.199 µg/mL and 98.22 µg/mL, and also inhibited tyrosinase with IC50 207.79 µg/mL and 221.51 µg/mL, respectively. The molecular docking proved that both compounds have affinities toward the enzymes.

Binahong (Anredera cordifolia (Tenore) Steen.) Leaf Extract Modulates Fatty Acids and Amino Acids to Lower Blood Glucose in High-Fat Diet-Induced Diabetes Mellitus Rats.

Patients with diabetes are 1.6 times more likely to use complementary alternative medicine than nondiabetic patients. Previous studies have shown that Anredera cordifolia (Tenore) Steen. (A. cordifolia) leaf extract has the capacity to lower blood glucose, but the actual mechanisms are unclear. Therefore, in this study, we explored the effect of A. cordifolia leaf extract on the metabolism of fatty acids and amino acids. Six-week-old male Wistar rats were randomly divided into six experimental groups (n = 5 per group). Two groups were fed with a regular diet or a high-fat diet (HFD) for six weeks. The regular diet and HFD groups were administered with 0.5% carboxymethylcellulose as a vehicle, and HFD rats were also fed with a suspension of glibenclamide (0.51 mg/kg body weight (BW)) or A. cordifolia leaf extract (25, 50, and 100 mg/kg BW). During the whole treatment, BW and food intake were recorded weekly. The rats were euthanized seven weeks after treatment. Blood glucose was evaluated by spectrophotometry, while fatty acids and amino acids were evaluated using a gas chromatography/flame ionization detector (GC/FID). All doses of A. cordifolia administration reduced blood glucose significantly, and 50 mg/kg BW was most effective in lowering blood glucose, similar to the effects of glibenclamide. A. cordifolia leaf extract affected the levels of medium-chain fatty acids, especially at 50 mg/kg BW. In contrast, glibenclamide affected long-chain fatty acids (LCFAs) to lower blood glucose. Based on the analysis conducted, we conclude that administration of A. cordifolia leaf extract can decrease blood glucose levels by regulating fatty acid metabolism and that a dose of 50 mg/kg BW in rats was the optimal dose.

Antielastase Activity of Macassar Kernels (Rhus javanica) Stem Extract and Skin Elasticity Evaluation of Its Topical Gel Formulation.

BACKGROUND: Macassar kernels (Rhus javanica L.) has potential as an antiaging agent as it has antielastase activity, especially its stem extract which has best percent inhibition compared to its leaves and fruit extract. Moreover, the antiaging agent can be commonly used in the form of gel for topical applications. Hence, formulation of HEC-based topical gel from the stem extract of Macassar kernels was conducted. This study aims to determine the antielastase activity of the stem extract of Macassar kernels and evaluate the skin elasticity of its topical gel formulation by conducting dermatological safety and skin antiaging efficacy test. METHODS: The stem extract was in vitro tested for antielastase activity using a microplate reader. Then, a formulation of a topical gel containing Rhus javanica stem extract was made. Five stages of quality control, which consisted of an organoleptic test, homogeneity test, pH measurement, viscosity measurement, and physicochemical stability test, were conducted to ensure the quality of topical gel formulation. Last, clinical studies were conducted to evaluate the dermatological safety and antiaging efficacy of gel preparation containing stem extract of Rhus javanica. Results. The stem extract provided antielastase activity (IC50 = 245.68 µg/mL), and its polyphenol was valued at 23.28 ± 1.52 mg GAE/g). The gel containing 10% stem extract had better stability than the gel containing 5% stem extract. The dermatology safety test and efficacy test results indicated that the topical gel containing 10% Rhus javanica stem extract did not cause any skin irritation and significantly improved skin elasticity (p < 0.05). In the treatment group, the moisture parameter was significantly changed on day 14 (p < 0.0001), day 21 (p < 0.0001), and day 29 (p < 0.0001). The elasticity parameter was also changed significantly on day 14 (p=0.0485), day 21 (p=0.0537), and day 29 (p=0.0002). CONCLUSION: The stem extract of Rhus javanica has potential antielastase activity. The topical gel containing Rhus javanica stem extract also has potential antielastase activity by increasing the skin moisture and enhancing skin elasticity.

Hypoglycemic Effect of a Combined Andrographis paniculata and Caesalpinia sappan Extract in Streptozocin-Induced Diabetic Rats.

INTRODUCTION: Researchers usually use herbal combinations to explore and develop traditional medicine to obtain additional benefits in the treatment of diseases, including diabetes. This study aims to evaluate the hypoglycemic effect of the combination of Andrographis paniculata (Burm. f.) Wall ex Nees and Caesalpinia sappan Linn extract (APCSE) on diabetes-induced rats. There has not been sufficient research on this combination; however, single extract studies of these plants have been widely conducted. MATERIALS AND METHODS: Male Sprague Dawley rats (160-200 g) were induced by injecting a low dose of streptozotocin (35 mg/kg BW) twice and fed with a high-fat diet containing 25% fat, whereas control animals received only standard feed. Rats were treated with APCSE at doses of 100 mg and 200 mg/kg BW for seven days and compared to the APE and CSE groups treated with the extract at 100 mg, respectively. For the control group, rats were treated with metformin with a dose of 250 mg/kg. The antihyperglycemic and antihyperlipidemic effects were determined by measuring blood glucose levels and lipid profiles (cholesterol, triglycerides, HDL, and LDL). To assess the impact of the extract on pancreatic and adipose tissue, the number of pancreatic beta cells and adipocytes was evaluated through histopathological and immunohistochemical study. Results and Discussion. In a nonfasting state, the blood glucose change in APCSE 200 mg was 18.65% and was significantly lower from the DM group. However, a single extract of APE and CSE showed lower fasting blood glucose levels compared to the combined extract. Lipid profiles show no significant differences in cholesterol levels between groups; however, all treatment groups, including metformin, showed higher triglyceride levels. The APE-treated group showed significantly lower HDL and LDL, whereas CSE only showed lower LDL. The ß-cell number was significantly higher after treatment with single extract CSE. The CSE and the combined extract groups showed hyperplasia adipocytes. CONCLUSION: The combined extract of APCSE has a moderate antihyperglycemic effect; however, a single extract may have better potential than the combined extract.

Inhibition of pancreatic elastase in silico and in vitro by Rubus rosifolius leaves extract and its constituents.

OBJECTIVE: Elastases are protease enzymes, which mainly hydrolyze proteins of the connective tissue, so they have a significant impact on human disease. Rubus rosifolius is one of the Rubus species found in Indonesian mountains, and it has potential as an elastase inhibitor. The objective of this research was to examine the in vitro elastase inhibitor activity of R. rosifolius leaves and to dock different ligands of its constituents against target protein of Porcine Pancreatic Elastase (PPE) receptor. METHOD: Dried leaves powder of R. rosifolius was extracted using Soxhlet apparatus with n-hexane, ethyl acetate, and methanol. The extract was evaporated, and in vitro elastase inhibitor activity was determined using PPE with the quercetin used as control positive. Selected nine constituents of R. rosifolius were evaluated on the docking behavior of elastase receptor using Protein-Ligand ANT System (PLANTS) computational software with PPE enzyme with Protein Data Bank (PDB) file 1BRU. RESULT: The methanol extract showed significantly inhibited elastase with IC50 186.13 µg/mL, but ethyl acetate extract showed weak activity, and n-hexane extract did not show any activity. Docking studies and binding free energy calculations and hydrogen bonding with some amino acids revealed that ellagic acid showed the least binding energy for the target enzyme. CONCLUSION: This research has opened new insights into understanding that constituents of R. rosifolius methanol extract are potential inhibitors against elastase, and suggested the active compound is ellagic acid.

The Antiviral Effect of Indonesian Medicinal Plant Extracts Against Dengue Virus In Vitro and In Silico.

Dengue infections are still a worldwide burden, especially in Indonesia. There is no specific medication against the dengue virus. Recently, many types of research have been conducted to discover a new drug for dengue virus using natural resource extracts. Indonesia, as a tropical country, has a wide biodiversity. There are several medicinal plants in Indonesia that are believed to possess anti-dengue activity, such as Myristica fatua, Cymbopogon citratus, and Acorus calamus plants. We conducted an in vitro laboratory experiment of several extracts from Indonesian herbs combined with in silico analysis. The extracts were evaluated for safety and antiviral activity in Huh7it-1 cell lines, using a single dose of 20 µg/mL and dose-dependent (5, 10, 20, 40, 80 and 160 µg/mL) of plant extracts against dengue virus serotype 2 (DENV-2) NGC strain. The DMSO 0.1% was used as a negative control. The cytotoxic aspect was assessed by counting the cell viability, while the antiviral activity was calculated by counting the average inhibition. The selectivity index (SI) of plant extracts were performed from a ratio of CC50/EC50 value. In silico analysis was conducted to determine the free energy of binding between NS5 of dengue virus with bioactive compounds contained in Myristica fatua, Cymbopogon citratus and Acorus calamus extract plants. We determined that all extracts were not toxic against Huh7it-1 cell lines. The methanolic extracts of A. calamus, C. citratus, and M. fatua showed inhibition of DENV-2 at a dose of 20 µg/mL to 96.5%, 98.9%, and 122.7%, respectively. The dose-dependent effects showed that M. fatua has the best inhibition activity towards DENV-2. Molecular docking result showed that artesunic acid within M. fatua has the best free energy of binding (-7.2 kcal/mol), followed by homoegonol (-7.1 kcal/mol) which was slightly different from artesunic acid among others. The methanolic extracts of A. calamus, C. citratus, and M. fatua showed prospective anti-dengue activities both in vitro and in silico. Future research should be conducted to find the pure extracts of all useful herbs as a new candidate of antiviral drug.

A new angiotensin-converting enzyme inhibitor from Peperomia pellucida (L.) Kunth

Objective: To isolate, identify, and evaluate a new angiotensin-converting enzyme inhibitor from Peperomia pellucida (L.) Kunth herbs. Methods: A dried sample of Peperomia pellucida herb was successively macerated with n-hexane and ethyl acetate. The ethyl acetate extract solution was evaporated to obtain the crude extract. Vacuum liquid column chromatography and thin layer chromatography were performed to obtain two pure compounds. Then, both compounds were elucidated and identified using the spectroscopic method. Angiotensin-converting enzyme inhibitory activity studies of both compounds were determined using angiotensin-converting enzyme kit WST-1 with spectrophotometer microplate reader 96-well at 450 nm wavelength. Results: Two bioactive compounds were successfully isolated from Peperomia pellucida herb, including a new compound of 2,3,5-trimethoxy-9-(12,14,15-trimethoxybenzyl)-1H-indene and pellucidin A. Both compounds demonstrated angiotensin-converting enzyme inhibitory activity, with IC50 values of 72 μM (27.95 μg/mL) and 11 μM (4.4 μg/mL), respectively. Conclusions: In the present study, two active angiotensin-converting enzyme inhibitors were successfully isolated and purified from Peperomia pellucida which is used as an antihypertensive in traditional medicine, and support its use as an angiotensin-converting enzyme-inhibiting drug.

α-Glucosidase Inhibitory Activity from Ethyl Acetate Extract of Antidesma bunius (L.) Spreng Stem Bark Containing Triterpenoids.

BACKGROUND: Buni (Antidesma bunius [L.] Spreng) has been used as a traditional antidiabetic agent in Asia. OBJECTIVE: The mechanism of antidiabetic properties was studied in this study by determine its α-glucosidase inhibitory activity. METHOD: Inhibition of α-glucosidase was performed in all fraction of Buni stem bark with acarbose and miglitol as standards. The half maximal inhibitory concentration (IC50) value of acarbose and miglitol was 5.75 and 59.76 µg/mL respectively while ethyl acetate (EtOAc) fraction was the most active fraction with IC50 of 19.33 µg/mL. Three isolates (B1, B2, and B3) were found in the EtOAc fraction and elucidated by infrared, 1hydrogen-nuclear magnetic resonance,13 carbon-nuclear magnetic resonance, and two-dimensional nuclear magnetic resonance. RESULT: The chemical structures of the isolates were identified by the spectrum then compared with literature which concluded that B1 is friedelin, B2 is ß-sitosterol, and B3 is betulinic acid. Inhibition of the α-glucosidase assay showed IC50 values of B1, B2, and B3 were 19.51, 49.85, and 18.49 µg/mL, respectively. SUMMARY: α-Glucosidase inhibitory activity assay was performed in n-hexane, ethyl acetate (EtOAc), methanol fraction of Buni (Antidesma bunius (L.) Spreng) stem bark and miglitolEtOAc fraction from the liquid chromatography has the highest inhibitory activity against α-glucosidaseThe chemical structures of the isolates were identified by the spectrums infrared, 1hydrogen-nuclear magnetic resonance,13 carbon-nuclear magnetic resonance, and two-dimensional nuclear magnetic resonance, then compared with literature which concluded that B1 is friedelin, B2 is ß-sitosterol, and B3 is betulinic acidBetulinic acid and friedelin showed the highest α-glucosidase inhibitory activity. Abbreviations used: IC50: Half maximal inhibitory concentration; H-NMR: Hydrogen-nuclear magnetic resonance; C-NMR: Carbon nuclear magnetic resonance; 2D-NMR: Two dimensional-nuclear magnetic resonance; EtOH: Ethanol; EtOAc: Ethyl acetate; MeOH: Methanol; CHCl3: Chloroform; DMSO: Dimethyl sulfoxide; EtF: Ethyl acetate fraction; Na2CO3: Sodium carbonate; IR: Infrared; TGR5: Transmembrane G protein-coupled receptor 5; EC50: Half maximal effective concentration.

Manzamine Alkaloids from an Acanthostrongylophora sp. Sponge.

Five new manzamine alkaloids (1-5) and new salt forms of two known manzamines (6 and 7), along with seven known compounds (8-14) of the same structural class, were isolated from an Indonesian Acanthostrongylophora sp. sponge. On the basis of the results of combined spectroscopic analyses, the structure of kepulauamine A (1) was determined to possess an unprecedented pyrrolizine moiety, while others were functional group variants of known manzamines. These compounds exhibited weak cytotoxicity, moderate antibacterial activity, and mild inhibition against the enzyme isocitrate lyase.

Antimicrobial activity of green tea extract against isolates of methicillin-resistant Staphylococcus aureus and multi-drug resistant Pseudomonas aeruginosa.

OBJECTIVE: To evaluate antibacterial activity of the Indonesian water soluble green tea extract, Camellia sinensis, against clinical isolates of methicillin-resistant Staphylococcus aureus (S. aureus) (MRSA) and multi-drug resistant Pseudomonas aeruginosa (MDR-P. aeruginosa). METHODS: Antimicrobial activity of green tea extract was determined by the disc diffusion method and the minimum inhibitory concentration (MIC) was determined by the twofold serial broth dilutions method. The tested bacteria using in this study were the standard strains and multi-drug resistant clinical isolates of S. aureus and P. aeruginosa, obtained from Laboratory of Clinical Microbiology, Faculty of Medicine, University of Indonesia. RESULTS: The results showed that the inhibition zone diameter of green tea extracts for S. aureus ATCC 25923 and MRSA were (18.970 ± 0.287) mm, and (19.130 ± 0.250) mm respectively. While the inhibition zone diameter for P. aeruginosa ATCC 27853 and MDR-P. aeruginosa were (17.550 ± 0.393) mm and (17.670 ± 0.398) mm respectively. The MIC of green tea extracts against S. aureus ATCC 25923 and MRSA were 400 µg/mL and 400 µg/mL, respectively, whereas the MIC for P. aeruginosa ATCC 27853 and MDR-P. aeruginosa were 800 µg/mL, and 800 µg/mL, respectively. CONCLUSIONS: Camellia sinensis leaves extract could be useful in combating emerging drug-resistance caused by MRSA and P. aeruginosa.