Review of the pharmacological properties of marine macroalgae used in the treatment of diabetes mellitus in Indonesia.

Indonesia is the largest archipelagic country in the world, with 70% of its territory covered by oceans that are rich in various types of biological resources. Indonesia's biodiversity has made it possible to develop natural medicine. Marine algae have enormous potential, but the types of marine algae used still need to be more varied. Research on the pharmacology of marine macroalgae has been conducted in Indonesia, but studies on such topic related to diabetes mellitus (DM) still need to be completed. This study provides a comprehensive dataset of pharmacological anti-diabetic potential of marine macroalgae used for managing DM and reports on preclinical trials that provide pharmacological evidence. Data on the Indonesian marine macroalgae used to lower blood glucose were obtained from online sources. The bioactive chemicals of marine macroalgae have been found efficient at blocking several diabetes enzymes in in-vivo and in-vitro studies, and such chemicals have anti-inflammatory, anti-obesity, antioxidant, and other therapeutic benefits. The Google Scholar was used to search for the pharmacological literature with the keywords marine AND macroalgae AND diabetes AND Indonesia. Pharmacological research on the anti-diabetic activity of marine macroalgae has been carried out on five major Indonesian islands, including Sumatra, Kalimantan, Java, Sulawesi, and Papua, which encompassed 12 provinces: Southwest Papua, South Sulawesi, West Kalimantan, Riau Archipelago, Banten, West Java, North Sulawesi, East Java, Yogyakarta, Maluku, Jakarta, and Bengkulu. Articles on preclinical tests (in vitro and in vivo) were also used for the phytochemical problem section. The results briefly describe which class of algae has been widely used in Indonesia as an anti-diabetic. The findings of this research can be utilized to help find DM treatment drugs based on natural resources from marine macroalgae.

Pharmacokinetic Herb-Drug Interactions of Glipizide with Andrographis paniculata (Burm. f.) and Andrographolide in Normal and Diabetic Rats by Validated HPLC Method.

Co-administered medicinal herbs can modify a drug's pharmacokinetics (PK), effectiveness, and toxicity. Andrographis paniculata (Burm. f.) ethanolic extract (APE) and andrographolide (AND) (a potent CYP2C9 inducer/inhibitor) can alter the pharmacokinetic parameters of glipizide (GLZ). This study aimed to determine the potential pharmacokinetics of herb−drug interactions between GLZ and APE/AND in the plasma of normal and diabetic rats using the HPLC bioanalysis method. The glipizide bioanalytical method established with RP-HPLC/UV instrument was validated following the EMA guidelines. GLZ was administered alone and in combination with APE or AND to normal and diabetic rats. The GLZ pharmacokinetic parameters were estimated according to the correlation between concentration and sampling time using the PK solver program. A simple and rapid GLZ bioanalysis technique with a lower limit of quantitation of 25 ng/mL was developed and presented the following parameters: accuracy (error ≤ 15%), precision (CV ≤ 15%), selectivity, stability, and linearity (R2 = 0.998) at concentrations ranging 25−1500 ng/mL. APE administration significantly improved the Cmax and AUC0−t/AUC0−∞ GLZ values in normal and diabetic rats (p < 0.05). AND significantly reduced the bioavailability of GLZ in diabetic rats with small values of T 1/2, Cmax, and AUC0−t/AUC0−∞ (p < 0.05). This combination can be considered in administering medications because it can influence the pharmacological effects of GLZ.

Designing and collecting data for a longitudinal study: the Sleman Health and Demographic Surveillance System (HDSS).

BACKGROUND: This paper describes the methodological considerations of developing an urban Health and Demographic Surveillance System (HDSS), in the Sleman District of Yogyakarta, Indonesia. METHODS: 1) The Sleman District was selected because it is mostly an urban area. 2) The minimum sample size was calculated to measure infant mortality as the key variable and resulted in a sample of 4942 households. A two-stage cluster sampling procedure with probability proportionate to size was applied; first, 216 Censuses Blocks (CBs) were selected, and second, 25 households in each CB were selected. 3) A baseline survey was started in 2015, and collected data on demographic and economic characteristics and verbal autopsy (VA); the 2nd cycle collected updated demographic data, VA, type of morbidity (communicable and non-communicable diseases, disability and injury) and health access. 4) The data were collected at a home visit through a Computer-Assisted Personal Interview (CAPI) on a tablet device, and the data were transferred to the server through the Internet. 5) The quality control consisted of spot-checks of 5% of interviews to control for adherence to the protocol, re-checks to ensure the validity of the interview, and computer-based data cleaning. 6) A utilization system was designed for policy-makers (government) and researchers. RESULTS: In total, 5147 households participated in the baseline assessment in 2015, and 4996 households participated in the second cycle in 2016 (97.0% response rate). CONCLUSIONS: Development of an urban HDSS is possible and is beneficial in providing data complementary to the existing demographic and health information system at local, national and global levels.

MALARIA SELF-CARE IN NIMBORAN SUBDISTRICT, JAYAPURA DISTRICT, PAPUA PROVINCE, INDONESIA.

Papua is one of five provinces with high malaria incidence in Indonesia. In 2009, the Indonesian Ministry of Health issued decree No 293 on malaria elimination. Socioeconomic, culture and psychological conditions, and perception of malaria are determining factors in seeking treatment. Health seeking behavior also are influenced by enabling factors, such as income and health insurance; and by health providers, such as availability of health care facilities, tariffs and living locationss. Self-care is one form of community participation in knowledge, prevention and early detection of malaria, and in seeking treatment and compliance to malaria treatment, especially among inhabitants in malaria endemic areas. This study was an observation in Nimboran Subdistrict, Jayapura District, Papua Province, Indonesia during 2013. Thirty individuals from 'Pengurus Rukun Tetangga' group were chosen randomly for the survey. Facts evaluated were knowledge of cause of malaria, disease transmission, symptoms and complication, diagnosis, treatment and side effects, medical seeking behavior and treatment, vector breeding sites, and attitude towards compliance of malaria treatment and use of mosquito nets. Self-care against malaria was considered important by 65% of the respondents. All participants had visited health centers and complied with prescribed drug regimen. All respondents with malaria-infected neighbors visited health centers. Regarding antimalarial malaria drugs, the majority of respondents knew of Darplex® and were aware that a common side effect of antimalarials was tinnitus. The majority of respondents identified ponds as malaria vector breeding places and recognized the importance of managing vectors in malaria prevention. The study concludes that malaria self-care was needed for awareness, prevention and treatment of this debilitating disease.

Diagnosis and Management of Infections Caused by Enterobacteriaceae Producing Extended-Spectrum b-Lactamase.

Bacterial resistance to antibiotics is a serious problem worldwide that affect the increment of morbidity and mortality rate; Enterobacteriaceae producing ESBL is one of the causes. However, there are still limited information regarding diagnosis and management of ESBL-E infection. Detection of ESBL-E requires certain steps that are problematic and time consuming. Diagnosis and management of ESBL-E infection have become more and more challenging due to limited diagnostic method available and choice of antibiotics that may be used, along with growing subtyped of ESBL through various of mutations. This article is aimed to give an overview on current situation of ESBL-E infections, with a focus on diagnosis and management of such infection by reviewing several recent studies on related issue.

Anti-inflammatory Activity of Saxifragin via Inhibition of NF-κB Involves Caspase-1 Activation.

Saxifragin, the 5-glucoside of the flavonoid quercetin, is found in plants and insects. It has been reported that saxifragin has peroxynitrite-scavenging effects. However, the mechanism of anti-inflammatory effects of saxifragin has not yet been clearly identified. In this study, we investigated the anti-inflammatory effects of saxifragin in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages and animal models of inflammation. We found that saxifragin suppressed the production of nitric oxide (NO) and prostaglandin E2 (PGE2) in LPS-activated RAW 264.7 macrophages by suppressing the level of protein and mRNA expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), respectively. Furthermore, saxifragin inhibited mRNA expression of pro-inflammatory cytokines including tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1ß. We studied the inhibitory effects of saxifragin on the nuclear translocation of nuclear factor (NF)-κB, activation of caspase-1, and phosphorylation of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK). Furthermore, pretreatment with saxifragin increased the survival rate of mice with LPS-induced septic death. Collectively, these findings suggest that saxifragin exerts anti-inflammatory activity by inhibiting NF-κB, caspase-1, and mitogen-activated protein kinase (MAPK) activation.

Effect of Chicoric Acid on Mast Cell-Mediated Allergic Inflammation in Vitro and in Vivo.

Chicoric acid (dicaffeoyl-tartaric acid), is a natural phenolic compound found in a number of plants, such as chicory (Cichorium intybus) and Echinacea (Echinacea purpurea), which possesses antioxidant, anti-inflammatory, antiviral, and analgesic activities. Although these biological effects of chicoric acid have been investigated, there are no reports of its antiallergic-related anti-inflammatory effects in human mast cells (HMC)-1 or anaphylactic activity in a mouse model. Therefore, we investigated the antiallergic-related anti-inflammatory effect of chicoric acid and its underlying mechanisms of action using phorbol-12-myristate 13-acetate plus calcium ionophore A23187 (PMACI)-stimulated HMC-1 cells. Chicoric acid decreased the mRNA expression of pro-inflammatory cytokines, such as tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1ß. We studied the inhibitory effects of chicoric acid on the nuclear translocation of nuclear factor kappa B (NF-κB) and activation of caspase-1. However, mitogen-activated protein kinase (MAPK) activation was not sufficient to abrogate the stimulus. In addition, we investigated the ability of chicoric acid to inhibit compound 48/80-induced systemic anaphylaxis in vivo. Oral administration of chicoric acid at 20 mg/kg inhibited histamine release and protected mice against compound 48/80-induced anaphylactic mortality. These results suggest that chicoric acid has an antiallergic-related anti-inflammatory effect that involves modulating mast cell-mediated allergic responses. Therefore, chicoric acid could be an efficacious agent for allergy-related inflammatory disorders.

Isoeugenin, a Novel Nitric Oxide Synthase Inhibitor Isolated from the Rhizomes of Imperata cylindrica.

Phytochemical studies on the constituents of the rhizomes of Imperata cylindrica (Gramineae) were performed using high-performance liquid chromatography (HPLC). We also aimed to search for any biologically active substance capable of inhibiting nitric oxide (NO) formation in lipopolysaccharide (LPS)-activated macrophage 264.7 cells, by testing four compounds isolated from this plant. Four compounds, including a new chromone, isoeugenin, along with ferulic acid, p-coumaric acid, and caffeic acid were isolated and identified by NMR spectroscopy. The structure of isoeugenin was determined as 7-hydroxy-5-methoxy-2-methylchromone by the 2D-NMR technique. Among the four compounds, isoeugenin has the lowest IC50 value on the inhibition of NO production in LPS-activated macrophage RAW264.7 cells (IC50, 9.33 µg/mL). In addition, isoeugenin significantly suppressed the LPS-induced expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and proinflammatory cytokines mRNA levels. Taken together, these results suggest that the anti-inflammatory activity of isoeugenin is associated with the down-regulation of iNOS, COX-2, and pro-inflammatory cytokines in RAW264.7 cells. Accordingly, our results suggest that the new chromone isoegenin should be considered a potential treatment for inflammatory disease.

Simultaneous quantification and validation of new peroxynitrite scavengers from Artemisia iwayomogi.

CONTEXT: Artemisia iwayomogi Kitamura (Compositae) has been very widely used for the treatment of acute or chronic hepatitis, jaundice, and gastritis. In the course of our continuing efforts to identify and quantify peroxynitrite scavengers from Compositae plants, A. iwayomogi was used in this study. OBJECTIVE: The present study was aimed to identify and quantify the peroxynitrite scavengers of A. iwayomogi. MATERIALS AND METHODS: Silica gel and ODS were used for column chromatography. The isolated compounds were quantified using an HPLC equipped with a Capcell Pak C18 column (5 µm, 250 mm × 4.6 mm i.d.), and the method was validated for the quality control. Peroxynitrite (ONOO(-))-scavenging activities of the compounds and extracts were evaluated on the measurement of highly fluorescent rhodamine 123 converted from non-fluorescent dihydrorhodamine (DHR)-123 under the presence of peroxynitrite. RESULTS: Based on the spectroscopic evidences, a new compound, 2"-O-caffeoylrutin (2"-O-trans-caffeic acid ester of quercetin 3-O-α-L-rhamnopyranosyl(1 → 6)-ß-D-glucopyranoside) was isolated and determined together with patuletin 3-O-glucoside, scopolin, scopoletin, rutin, 3,4-dicaffeoylquinic acid, and chlorogenic acid. All of them were potent peroxynitrite scavengers (IC50 ≤ 1.88 µg/mL). DISCUSSION AND CONCLUSION: The peroxynitrite scavengers were mainly distributed in the EtOAc fraction rather than the ether and BuOH fractions. The 70% MeOH extract exhibited a high peroxynitrite-scavenging activity. Through the validation, the present HPLC method was verified to be sufficiently sensitive, accurate, precise, and stable. Therefore, this method can be used for the quality control of A. iwayomogi.

Pharmacokinetics evaluation of newly formulated beads alginate/gum acacia loaded ketoconazole in rabbit plasma by oral administration.

Background and purporse: The combination of alginate and gum acacia in previous studies showed good results in inhibiting ketoconazole precipitation due to the supersaturation phenomenon. Ketoconazole-loaded alginate and gum acacia can produce hydrogel beads through cross-linking with Ca2+ using ionotropic gelation techniques. However, the pharmacokinetic study of the ketoconazole beads loaded to alginate and gum acacia needs further investigation. This study aimed to evaluate pharmacokinetic parameters using rabbits via oral administration. Experimental approach: The drug was administered orally to 2 groups of rabbits: pure ketoconazole (KTZ) and formulation of ketoconazole (AG75) groups. Blood samples were obtained from the ear marginal vein at various time points: 0 (before administration), 15, 30, 45, 60, 90, 120, 150, 180, 240, 300, 360, and 420 minutes after oral dosage. The pharmacokinetic study employed a non-compartment analysis to calculate the area under the curve (AUC), the volume of distribution (Vd F-1), clearance (Cl F-1), maximum concentration (Cmax), and time to reach maximum concentration (tmax). The data obtained from the parameter result was analyzed using the independent-sample T-test. Key result: The results of the KTZ group include AUC of 15.83±0.62 h µg mL-1, VdF-1 of 8.95±1.17 mL, ClF-1 of 3.45±0.3 mL h-1, Cmax of 4.7±0.69 µg mL-1, and tmax of 1.67±0.17 h. The results of the AG75 group include AUC of 27.8±1.01 h µg mL-1, VdF-1 of 11.5±2.4 mL, ClF-1 of 2.15±0.11 mL h-1, Cmax of 4.49±0.52 µg mL-1, and tmax of 2.5±0.5 h. Conclusion: The formulation incorporating ketoconazole beads resulted in a higher AUC0-∞ than the pure ketoconazole. This finding suggests that the created formulation has enhanced the bioavailability of ketoconazole.

A Wound-Healing Effect of Nanoemulgel of Kangkung (Ipomoea Reptans, Poir) Leaf Extract in STZ Diabetic Rats.

Context: The chronic diabetes mellitus (DM) condition may lead to diabetic wounds that increase morbidity in patients. Ipomoea reptans Poir leaves have been widely reported to possess anti-diabetic activity due to their flavonoid contents. To enhance drug penetration, a nanoemulgel preparation was formulated. Aims: This study aimed to evaluate the activity of nanoemulgel preparations of Ipomoea reptans Poir leaf extract on diabetic and non-diabetic wound-healing using male Wistar rats. Settings and Design: This research was an experimental study with a post-test only control group design. Materials and Methods: The rats (n = 32) were randomly divided into two groups: diabetic (induced by 40 mg/kg BW STZ) and non-diabetic model. Each model consisted of four groups: normal, positive control, I. reptans leaf extract (IRLE), and nanoemulgel of I. reptans leaf extract (NIRLE). All the animals studied were shaved from the back, and a 2.5 × 0.5 cm full-thickness excision wound was made. IRLE and NIRLE were administered daily and observed for the wound-healing process. Statistical Analysis Used: The one-way analysis of variance with the Tukey post-hoc test was used for the statistical analysis. Results: A NIRLE formulation has been developed to produce a preparation that meets the physical requirements. IRLE and NIRLE possessed wound-healing activity in normal and diabetic rat models. However, the wound-healing process in diabetic rats treated with NIRLE was faster than those with IRLE. Conclusions: NIRLE increased the activity of wound-healing effect of I. reptans leaves on diabetic rats in comparison with the extract form.

Lessons Learned From Telephone-Based Data Collection for Health and Demographic Surveillance Systems During the COVID-19 Pandemic in Indonesia.

The Sleman Health and Demographic Surveillance System (HDSS) is a longitudinal survey held routinely since 2014 to collect demographic, social, and health changes in Sleman Regency, Special Region of Yogyakarta, Indonesia. During the COVID-19 pandemic in Indonesia, we needed to adjust our method of conducting data collection from in-person to telephone interviews. We describe the Sleman HDSS data collection strategy used and the opportunities it presented. First, the Sleman HDSS team completed a feasibility study and adjusted the standard operational procedures to conduct telephone interviews. Then, the Sleman HDSS team collected data via a telephone interview in September-October 2020. Ten interviewers were equipped with an e-HDSS data collection application installed on an Android-based tablet to collect data. The sample targeted was 5,064 households. The telephone-based data collection successfully interviewed 1,674 households (33% response rate) in 17 subdistricts. We changed the data collection strategy so that the Sleman HDSS could still be conducted and we could get the latest data from the population. Compared to in-person interviewing, data collection via telephone was sufficiently practical. The telephone interview was a safe and viable data collection method. To increase the response rate, telephone number activation could be checked, ways of building rapport could be improved, and engagement could be improved by using social capital.

Anti-diabetic effect of a combination of andrographolide-enriched extract of Andrographis paniculata (Burm f.) Nees and asiaticoside-enriched extract of Centella asiatica L. in high fructose-fat fed rats.

Traditionally, a combination of medicinal plants is commonly used for lowering blood glucose in diabetic patients in order to provide additional benefits of the single drug. A. paniculata and C. asiatica are two traditional medicines form South Asian and Southeast Asain countries consumed by people for treating daibates mellitus and its complications. Hyperglycemia in the rats was stimulated by high fructose-fat diet that consists of 36% fructose, 15% lard, and 5% egg yolks in 0.36 g/200 g body weight for 70 days. The rats were orally administered with the combination of andrographolide-enriched extract of A. paniculata (AEEAP) leaves and asiaticoside-enriched extract of C. asiatica (AEECA) herbs from day 70 for 7 days. Antidiabetic activity was evaluated by estimating mainly the blood glucose levels and other parameters such as HDL, LDL, cholesterol and triglyceride. The results showed that combination at the ratio of 70:30 exhibited a promosing antidiabetic effect in high-fat-fructose-fed rat, and exhibited sinergistic effects on blood cholesterol and HDL levels. It can be concluded that its antidiabetic effect was better than that of single treatment of AEEAP or AEECA. That combination was also potential to develop as a blood glucose-lowering agent for diabetic patients.

Antiangiogenic Activity of n-hexane Insoluble Fraction and Its Tylophorine Component from Ficus septica Leaves in Chicken Chorioallantoic Membrane Induced by bFGF.

OBJECTIVE: Ficus septica is an Indonesian medicinal plant traditionally used to treat various illness, including cancer. The n-hexane insoluble fraction of the ethanolic extract of F. septica leaves (HIFFS) shows a potential anticancer activity against breast cancer cell line T47D. Considering that angiogenesis is a pivotal factor in malignant cancer growth, progression, and invasion, we aimed to investigate the antiangiogenic effect of HIFFS on chicken chorioallantoic membrane (CAM) induced by bFGF. We also evaluated tylophorine, the cytotoxic alkaloid of F. septica. METHODS: Chicken CAM was used to assess the antiangiogenic effect. Fertilized chicken eggs were induced with basic fibroblast growth factor (bFGF) ex ovo. Prior to bFGF induction, HIFFS (2.33, 4.65, 6.98, and 9.30 µg/mL) or tylophorine (9.20 µM) was added (10 µL) to a paper disk and implanted to the CAM. After 48 h of incubation, each treatment group was photographed, and the number of new blood vessel was calculated and compared with that in the solvent-treated group to determine the antiangiogenic activity. Histology of the CAM was evaluated after hematoxylin-eosin and Mallory acid fuchsin staining. RESULTS: We found that HIFFS at low concentrations (2.33, 4.65, 6.98, and 9.30 µg/mL) inhibited angiogenesis activity (31.87, 41.99, 53.65, and 70.08, respectively) in chicken CAM induced by bFGF. Tylophorine (9.20 µM) also showed similar antiangiogenesis activity in the same model. Histopathology analysis revealed that HIFFS and tylophorine reduced the number of new blood vessels in CAM induced by bFGF. CONCLUSION: HIFFS and tylophorine showed antiangiogenic effect on chicken CAM induced by bFGF. This finding emphasized the potential of F. septica as a candidate anticancer agent.

Regulation of mucosal mast cell activation by short interfering RNAs targeting syntaxin4.

Mucosal mast cells (MMCs) have an important role in allergic inflammation, and effective antagonists are required for their regulation. To discover a possible mechanism of controlling the activation of MMCs, we investigated the expression and function of syntaxin4, one of the soluble membrane N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins, in RBL-2H3 cells, which is a rat mucosal mast cell line. Syntaxin4 silencing was induced by transfection of short interfering RNAs (siRNAs). Syntaxin4 was knocked down in mast cells at both the mRNA and protein levels. The release of granule contents that are involved in inflammation, such as histamine and hexosaminidase, was significantly suppressed by the gene silencing of syntaxin4. Silencing of this gene was also induced in the trachea and bronchi of rats by intratracheal application of the siRNAs using an atelocollagen delivery system. The activation of MMCs, which was monitored by the level of rat mast cell protease-II (RMCPII) in the bronchoalveolar lavage fluid (BALF), was inhibited, and asthmatic airway constriction was prevented by administration of the syntaxin/atelocollagen complex. These results indicate that siRNAs targeting syntaxin4 can stabilize mucosal mast cells and may have beneficial therapeutic effects on the asthmatic response.

Characterization of pyrene biodegradation by white-rot fungus Polyporus sp. S133.

A white-rot fungus of Polyporus sp. S133 was isolated from an oil-polluted soil. The metabolism of pyrene by this fungus was investigated in liquid medium with 5 mg of the compound. Depletion of pyrene was evident during the 30-day growth period and was 21% and 90%, respectively, in cometabolism and metabolism of pyrene alone. Pyrene was absorbed to fungal cells or biodegraded to form simpler structural compounds. Seventy-one percent of eliminated pyrene was transformed by Polyporus sp. S133 into other compounds, whereas only 18% was absorbed in the fungal cell. The effects of pH and temperature on biomass production of Polyporus sp. S133 for pyrene were examined; the properties of laccase and 1,2-dioxygenase produced by Polyporus sp. S133 during pyrene degradation were investigated. The optimal values of pH were 3, 5, and 4 for laccase, 1,2-dioxygenase, and biomass production, respectively, whereas the optimal values of temperature were 25 °C for laccase and 50 °C for 1,2-dioxygenase and biomass production. Under optimal conditions, pyrene was mainly metabolized to 1-hydroxypyrene and gentisic acid. The structure of 1-hydroxypyrene and gentisic acid was determined by gas chromatography-mass spectrometry after identification using thin-layer chromatography.

Synergistic effects of ethyl acetate fraction of Ficus septica Burm. f. and doxorubicin chemotherapy on T47D human breast cancer cell line.

OBJECTIVE: Previously, ethanolic extract of Ficus septica Burm. f. (Moraceae) leaves and its ethyl acetate soluble fraction (EASF) exhibited potent cytotoxic effects on T47D breast cancer cells. In the present study, we further investigated the effects of EASF of ethanolic extract of F. septica leaves in combination with doxorubicin on T47D breast cancer cell line in cytotoxicity, cell cycle arrest and apoptosis induction. METHODS: The cytotoxic effect analysis on T47D cells was carried out using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Analysis of cell cycle distribution was performed using a flowcytometer and the data were analyzed using ModFit LT 3.0 program. Apoptosis assay was carried out by double staining method using ethidium bromide-acridin orange. The expression of cleaved-poly ADP-ribose polymerase in the T47D cell line was identified using immunohistochemical techniques. RESULTS: The combination of doxorubicin (2 to 8 nmol/L) with EASF (0.875 to 7 µg/mL) more potently inhibited cell growth than the single treatment of doxorubicin in T47D cells. In addition, the combination of doxorubicin and EASF could increase the incidence of cells undergoing apoptosis. EASF was found to improve cytotoxic effect of doxorubicin by changing the inhibition of cell cycle G(2)/M to G(1) phase. The combination also exhibited a more intensive stimulatory effect on cleaved-PARP expression in T47D cells than the single treatment. CONCLUSION: It is concluded that EASF may enhance doxorubicin activities in T47D cells by inducing apoptosis and cell cycle arrest.

Clitorin ameliorates western diet-induced hepatic steatosis by regulating lipogenesis and fatty acid oxidation in vivo and in vitro.

Nonalcoholic fatty liver disease (NAFLD) is usually correlated with metabolic diseases, such as obesity, insulin resistance, and hyperglycemia. Herein, we investigated the inhibitory effects and underlying governing mechanism of clitorin in a western diet (WD)-induced hepatic steatosis mouse model, and in oleic acid-stimulated HepG2 cells. Male C57BL/6 mice were fed a normal diet, WD, WD + 10 or 20 mg/kg orlistat, and WD + 10 or 20 mg/kg clitorin. HepG2 cells were treated with 1 mM oleic acid to induce lipid accumulation with or without clitorin. Clitorin significantly alleviated body weight gain and hepatic steatosis features (NAFLD activity score, micro-, and macro-vesicular steatosis) in WD-induced hepatic steatosis mice. Additionally, clitorin significantly decreased protein expressions of sterol regulatory element-binding protein 1 (SREBP1), peroxisome proliferator-activated receptor γ (PPARγ), and CCAAT/enhancer binding protein α (C/EBPα) in WD-induced hepatic steatosis mice. Moreover, clitorin significantly diminished the mRNA levels of SREBP1, acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), and hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR) and enhanced the mRNA levels of peroxisome proliferator-activated receptor α (PPARα) and carnitine palmitoyltranserase-1 (CTP-1), as well as adenosine monophosphate-activated protein kinase (AMPK) in the liver of WD-induced hepatic steatosis mice and oleic acid-stimulated HepG2 cells. Overall, our findings demonstrated that clitorin can be a potentially efficacious candidate for NAFLD management.

Effects of aegeline, a main alkaloid of Aegle Marmelos Correa leaves, on the histamine release from mast cells.

Aegeline or N-[2-hydroxy-2(4-methoxyphenyl) ethyl]-3-phenyl-2-propenamide is a main alkaloid isolated from Aegle marmelos Correa collected in Yogyakarta Indonesia. In our study, we investigated the effects of aegeline on the histamine release from mast cell. The study was performed by using (1) rat basophilic leukemia (RBL-2H3) cell line, and (2) rat peritoneal mast cells (RPMCs). DNP(24)-BSA, thapsigargin, ionomycin, compound 48/80 and PMA were used as inducers for histamine release from mast cell. In our study, aegeline inhibited the histamine release from RBL-2H3 cells induced by DNP(24)-BSA. Indeed, aegeline showed strong inhibition when RBL-2H3 cells induced by Ca(2+) stimulants such as thapsigargin and ionomycin. Aegeline is suggested to influence the intracellular Ca(2+) pool only since could not inhibit the (45)Ca(2+) influx into RBL-2H3 cells. Aegeline showed weak inhibitory effects on the histamine release from RPMCs, even though still succeed to inhibit when the histamine release induced by thapsigargin. These findings indicate that aegeline altered the signaling pathway related to the intracellular Ca(2+) pool in which thapsigargin acts. Based on the results, the inhibitory effects of aegeline on the histamine release from mast cells depended on the type of mast cell and also involved some mechanisms related to intracellular Ca(2+) signaling events via the same target of the action of thapsigargin or downstream process of intracellular Ca(2+) signaling in mast cells.

Effects of marmin, a compound isolated from Aegle marmelos Correa, on contraction of the guinea pig-isolated trachea.

Marmin or 7-(6',7'-dihydroxygeranyl-oxy)coumarin is a compound isolated from Aegle marmelos Correa. In the study, we examined the effects of marmin on the contraction of guinea pig-isolated trachea stimulated by several inducers, namely histamine, metacholine, compound 48/80. We also evaluated its action against contraction induced by extracellular or intracellular calcium ion. The possibility of marmin to potentiate the relaxation effect of isoprenaline was also studied. Marmin added in the organ bath at 10 min prior to the agonist inhibited the contraction elicited by histamine and metacholine in a concentration-dependent manner. Moreover, marmin antagonized the histamine-induced contraction in competitive manner. Marmin mildly potentiated the relaxation effect of isoprenaline. In the study, marmin abrogated the contraction of tracheal smooth muscle induced by compound 48/80, an inducer of histamine release. Besides, marmin successfully inhibited CaCl(2)-induced contraction in Ca(2+)-free Krebs solution. Marmin also inhibited two phases of contraction which were consecutively induced by metacholine and CaCl(2) in Ca(2+)-free Krebs solution. Based on the results we concluded that marmin could inhibit contraction of the guinea-pig tracheal smooth muscle, especially by interfering histamine receptor, inhibiting the histamine release from mast, inhibiting intracellular Ca(2+) release from the intracellular store and the Ca(2+) influx through voltage-dependent Ca(2+) channels.