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Three water soluble dicopper(II) complexes, tetrakis(μ-4-chloro-2-nitrobenzoato-κ2O,O′) bis[aquacopper(II)] (1), tetrakis(μ-2-chloro-6-fluorobenzoato-κ2O,O′)bis[aquacopper(II)] (2) and tetrakis(μ-2-chlorobenzoato-κ2O,O′)bis[aquacopper(II)] (3), have been synthesized. Complex 1 crystallizes in monoclinic system, space group C2/c with a = 28.813(3), b = 7.2368(6), c = 17.0053(14) A, β = 105.395° and Z = 4, while 2 crystallizes in triclinic system, space group P 1 ¯ with a = 7.5638(2), b = 13.0494(4), c = 16.7079(5) A, α = 106.003°, β = 90.388°, γ = 91.855° and Z = 2. In addition, all the complexes have been characterized by their elemental analysis, FT-IR, UV–Vis spectroscopy and cyclic voltammetry. All complexes have been found to be redox active displaying two quasi-reversible redox couples corresponding to CuIICuII to CuIICuI and CuIICuI to CuICuI redox processes, respectively. The results from the gel electrophoresis experiment of 1–3 on pBR322 DNA, have demonstrated the occurrence of oxidative cleavage in the presence of hydrogen peroxide. The hydroxyl radical has been found to be the reactive oxygen species that is responsible for the cleavage reaction. In the antiproliferation tests against MCF-7, DU 145, HeLa and HepG2 cancer cell lines, all complexes show significant preference against HepG2 cell lines. All complexes also show significant preference against HepG2 cancer cells line over Chang human normal liver cells which means 1–3 are more potent against cancer cells than normal cells. In antibacterial tests against nine pathogenic bacterial species, all complexes show a very selective trend whereby they exhibit antibacterial activity against the Gram negative bacteria Enterobacter aerogenes.

Kidney stone disease is a common health problem in industrialised nations. We developed a SDS-PAGE-based method to quantify Tamm Horsfall glycoprotein (THP) for screening of kidney stone disease. Urinary proteins were extracted by using ammonium sulphate precipitation at 0.27 g salt/mL urine. The resulted pellet was dissolved in TSE buffer. Ten microliters of the urinary proteins extract was loaded and separated on 10% SDS-PAGE under reducing condition. THP migrated as single band in SDS-PAGE. The assay reproducibility and repeatability were 4.8% CV and 2.6% CV, respectively. A total of 117 healthy subjects and 58 stone patients were tested using this assay, and a distinct cut-off (P < 0.05) at 5.6 μg/mL THP concentration was used to distinguish stone patients from healthy subjects. The sensitivity and specificity of the method were 92.3% and 83.3%, respectively.

The acute toxicity of standardized extract of Orthosiphon stamineus was studied in Sprague Dawley rats. The rats were administered a single dose of 5000 mg/kg body weight (BW) orally on Day 0 and observed for 14 days. There were no deaths recorded and the animals did not show signs of toxicity during the experimental period. The effect of the extract on general behavior, BW, food and water intake, relative organ weight per 100 g BW, hematology and clinical biochemistry were measured. All the parameters measured were unaffected as compared to the control. The acute toxicity LD50 was estimated to be >5000 mg/kg BW.

Renal calculi disease or known as kidney stone disease is the most common urological disorder in both men and women, although it is more prevalent in men. The lifetime chance for an individual to develop renal calculi is approximately 10% whereas the risk of recurrence in a 10-year period is 74%. Therefore, a diagnostic tool for screening or detecting renal calculi is greatly needed. In this study, we analyze urinary protein profiles from patients with renal calculi for the first time (RC), healthy subjects (HS), and patients with recurrent renal calculi (RRC) to identify a biomarker for detecting the disease. Urinary proteins were isolated by salt precipitation and the proteins resolved by SDS-PAGE. Target proteins were analyzed with LC/MS/MS. Thirty-two proteins were identified from healthy subjects and patients. Uromodulin was the most abundant urinary protein in HS but was a very faint band if detected at all from those that formed renal calculi for the first time (p0.05). Yet the excreted levels of urinary uromodulin in RRC were similar to those of the HS suggesting that uromodulin is a reliable biomarker for only RC. In addition, a few immunoglobulins that were commonly found in the urine of both RC and RRC, which include Ig alpha heavy chain 1, Ig gamma-2 c region, Ig gamma-3 heavy chain disease protein, Ig heavy chain variable region, Ig heavy constant region gamma 4, and Ig heavy chain. Ig heavy chain Fab frag and antibody a5b7 chain B may serve as potential biomarkers for renal calculi disease.

The hydrodistilled essential leaves and stems oils of Orthosiphon stamineus Benth were analysed by GC–MS/MS. Sixty nine compounds representing 97.6 and 97.4% of the total leaves and stems oils, respectively were identified, of which -caryophyllene (24.0 and 35.1%), -humulene (14.2 and 18.4%), -elemene (11.1 and 8.5%), 1-octen-3-ol (8.2 and 7.0%), bourbonene (3.4 and 3.0%), -pinene (2.1 and 1.7%), caryophyllene oxide (1.6 and 2.2%), camphene (1.6 and 1.3%) and limonene (1.2 and 1.1%) were the major compounds. Thus, the monoterpenes and sesquiterpenes were the predominant portions of the oils. Essential oils and methanol extract of O. stamineus and the derived fractions of hexane, chloroform, and ethyl acetate were tested for anti-fungal activity, which was determined by disc diffusion and minimum inhibitory concentration (MIC) determination methods. The oils, methanol extract and derived fractions of methanol extract displayed great potential of anti-fungal activity as a mycelial growth inhibitor against the tested phytopathogenic fungi such as Botrytis cinerea, Rhizoctonia solani, Fusarium solani, Colletotricum capsici and Phytophthora capsici, in the range of 49.3–70.3% and minimum inhibitory concentration ranging from 500 to 1000g/ml.

A disposable screen printed array sensor strip based on self-plasticized lipid membranes combined with chemometric algorithm has been developed and applied for quantification of Orthosiphon stamineus Benth extracts. Sinensetin, a pharmacologically active flavonoid in Orthosiphon stamineus Benth, was quantified with the sensor system using standard addition method. The method was compared with high performance thin layer chromatography (HPTLC). Partial least square (PLS) and principal component regression (PCR) were applied to the array sensor output to determine the sinensetin in O. stamineus samples from different suppliers. Comparison between the PLS and PCR models presented in the quantitative analysis showed that PLS have substantially better predictive capability than PCR. The root mean square error (RMSE) of Prediction for PLS and PCR were 0.17 ppm and 0.19 ppm, respectively. The concentration of sinensetin by PLS fell within the range of 0.25%–0.30% in six different batches of extracts that were supplied by Hovid Sdn Bhd (HV) while a range 0.18%–0.24% was obtained in ten different batches of extracts supplied by Nusantara Herbs Sdn Bhd (NH). The array sensor showed good correlation (0.9902) with the HPTLC method.

Kidney stone disease affects 1 - 20% of the general population. At present, the diagnosis of a stone is done using radiography method when noticeable symptoms appeared. We developed a non-invasive quantitative assay for urinary THP, namely ELISA; whereby our previous study and other reports had shown the usefulness of THP as biomarker for kidney stone disease. Since urine is biological fluid that is easily obtainable, this method could be used as a screening assay for kidney stone prior to confirmation with radiography. The ELISA gave assay linearity r2 > 0.999 within the range of 109 ng/mL to 945 ng/mL THP. Assay precisions were < 4% (C.V.) for repeatability and < 5% (C.V.) for reproducibility. Assay accuracy range from 97.7% to 101.2% at the various THP concentrations tested. Assay specificity and sensitivity were 80% and 86%, respectively. The cut-off points at P < 0.05 were 37.0 and 41.2 μg/mL for male and female, respectively. The assay is cost effective and rapid whereby the cost for assaying each urine sample in duplicate is approximately USD0.35 and within 5 hours, 37 samples can be assayed alongside full range of standards and 3 QC samples in each plate. Furthermore, sample preparation is relatively easy where urine sample was diluted 10 times in TEA buffer. The usability of the ELISA method for diagnosis of kidney stone disease is evaluated with 117 healthy subjects and 58 stone formers.

Kidney stone disease affects 1 - 20% of the general population. At present, the diagnosis of a stone is done using radiography method when noticeable symptoms appeared. We developed a non-invasive quantitative assay for urinary THP, namely ELISA; whereby our previous study and other reports had shown the usefulness of THP as biomarker for kidney stone disease. Since urine is biological fluid that is easily obtainable, this method could be used as a screening assay for kidney stone prior to confirmation with radiography. The ELISA gave assay linearity r2 > 0.999 within the range of 109 ng/mL to 945 ng/mL THP. Assay precisions were < 4% (C.V.) for repeatability and < 5% (C.V.) for reproducibility. Assay accuracy range from 97.7% to 101.2% at the various THP concentrations tested. Assay specificity and sensitivity were 80% and 86%, respectively. The cut-off points at P < 0.05 were 37.0 and 41.2 μg/mL for male and female, respectively. The assay is cost effective and rapid whereby the cost for assaying each urine sample in duplicate is approximately USD0.35 and within 5 hours, 37 samples can be assayed alongside full range of standards and 3 QC samples in each plate. Furthermore, sample preparation is relatively easy where urine sample was diluted 10 times in TEA buffer. The usability of the ELISA method for diagnosis of kidney stone disease is evaluated with 117 healthy subjects and 58 stone formers.

Hibiscus sabdariffa tea is a widely used medicinal beverage and a treatment for high blood pressure and high blood cholesterol in many parts of the world. Many studies on H. sabdariffa have been conducted including extraction and identification of main biocompounds. However, information on the effects of processing the plant is scarce. This is important as sample processing procedure influence the composition of the end product. Hence, the main objective of this present study was to examine the effect of sample processing (non-extracted, ethanol extract and water extract) on H. sabdariffa composition. Fourier Transform Infrared (FTIR) was used for the process of identification. The powdered sample of H. sabdariffa (FT34) was obtained from a local company in Peninsula Malaysia. A fresh sample obtained from the same company was processed in the Phytochemistry Laboratory, Institute for Medical Research and labelled as FT35. Sample and potassium bromide (KBr) were mixed (1:250) to form a 1-2 mm transparent disk under 9.80 psi in vacuum. The FTIR Spectra were recorded with 32 scans and 0.2 cms-1 OPD speed. Spectra of FT34 and FT35 raw samples indicated obvious differences in the range of 1500-1135 cm-1. The FT34 ethanol extract using trifluoroacetic acid (TFA) showed that the peak at 1629 cm-1 was the highest in the range of 1800-1500 cm-1, whereas for FT35, the highest peak was 1739 cm-1. The peak at 1071 cm-1 of FT35 was the only one compatible to standard dephinidin-3-O-sambubioside and cyanidin-3-Osambubioside which are used for qualification of sample content. In fact, both standards showed up as different chromatographs in thin layer chromatography. Water extract of FT35 showed a peak at 1676 cm-1 which was not detected in water extract spectrum of FT34, while the pattern of spectrum varied within the range of 1300-400 cm-1. Second derivative spectra enhanced the comparable base peaks of both sample and the target standards. There were five matched ethanol extract base peaks, indicating the macrofingerprint of H. sabdariffa. Two dimensional correlation spectrum of FT34 raw powder showed different correlation spot especially in the cluster of 1425 cm-1 to 1743 cm-1 compared with FT35. The three-stage infrared spectroscopy comprehensively analysed the holographic spectra and hierarchically characterized the integrated constituents involved.

Background Orthosiphon stamineus (OS) Benth is a medicinal plant and native in Southeast Asia. Pharmacological effects of OS are attributed to the presence of lipophilic flavones. However; lipophilic compounds suffer from poor aqueous solubility which limits the OS oral bioavailability and therapeutic applications. Therefore, OS was prepared in nano formulation form using liposomes from soybean phospholipids. The aim of the present study is to evaluate the in vitro genotoxicity and in vivo oral toxicity of nano liposomes of OS ethanolic extract (OS-EL). Methods In the acute toxicity study Sprague Dawley female rats were given a single dose of the OS-EL at 5000 mg/kg/day orally and screened for two weeks after administration. In the subchronic study, three different doses of OS-EL were administered for 28 days. Mortality, clinical signs, body weight changes, hematological and biochemical parameters, gross findings, organ weights, and histological parameters were monitored during the study. Genotoxicity was assessed using the Ames test with the TA98 and TA100 Salmonella typhimurium strains. High-performance liquid chromatography was performed for identification and quantification of the major marker compounds in OS-EL. Heavy metal detection was performed using an atomic absorption spectrometer. Results The acute toxicity study showed that the LD50 of the extract was greater than 5000 mg/kg. In the repeated dose 28-day oral toxicity study, the administration of 250 mg/kg, 500 mg/kg, and 1000 mg/kg/day of OS-EL per body weight revealed no significant difference in food and water consumptions, bodyweight change, haematological and biochemical parameters, relative organ weights, gross findings or histopathology compared to the control group. The Ames test revealed that the OS-EL did not have any potential to induce gene mutations in S. Typhimurium. Conclusions Analyses of these results with the information of signs, behaviour, and health monitoring could lead to the conclusion that the long-term oral administration of OS-EL for 28 days does not cause sub-chronic toxicity.

Two flavanones named (2S)-7-Hydroxy-5-methoxy-6,8-dimethyl flavanone (1), (S)-5,7-dihydroxy-6,8-dimethyl-flavanone (2), along with known chalcone, namely, (E)-2ʹ,4ʹ- dihydroxy-6ʹ-methoxy-3ʹ,5ʹ-dimethylchalcone (3) and two triterpenoids, namely, betulinic and ursolic acids (4 and 5), were isolated from the leaves of Syzygium campanulatum Korth (Myrtaceae). The structures of compounds (1 and 2) were determined on the basis of UV-visible, FTIR, NMR spectroscopies and LC-EIMS analytical techniques. Furthermore, new, simple, precise, selective, accurate, highly sensitive, efficient and reproducible RP-HPLC method was developed and validated for the quantitative analysis of the compounds (1–5) from S. campanulatum plants of five different age. RP-HPLC method was validated in terms of specificity, linearity (r2 ≤ 0.999), precision (2.0% RSD), and recoveries (94.4%–105%). The LOD and LOQ of these compounds ranged from 0.13–0.38 and 0.10–2.23 μg·mL−1, OPEN ACCESS respectively. Anti-proliferative activity of isolated flavanones (1 and 2) and standardized extract of S. campanulatum was evaluated on human colon cancer (HCT 116) cell line. Compounds (1 and 2) and extract revealed potent and dose-dependent activity with IC50 67.6, 132.9 and 93.4 μg·mL−1, respectively. To the best of our knowledge, this is the first study on isolation, characterization, X-ray crystallographic analysis of compounds (1 and 2) and simultaneous RP-HPLC determination of five major compounds (1–5) from different age of S. campanulatum plants.

Background Studies on selected metabolites profiling of Orthosiphon stamineus extracts using chromatographic and spectroscopic techniques combined with chemometric tools have not been fully elucidated. Thus present study was performed to profile selected metabolites in O. stamineus leaves extracts using HPLC and FTIR combined with chemometric tools and correlated with biological activities. Methods Five different extracts were prepared using three methods; maceration, soxhlet and reflux. The extracts were analyzed using UV-Vis, HPLC and FTIR techniques. Analysis of selected primary and secondary metabolites was also evaluated. The antioxidant and cytotoxic activities of the extracts were evaluated. Chemometric tools were employed to classify the extracts based on HPLC analysis and FTIR fingerprints. Results The ethanolic extract using maceration characterized high content of phenolics and flavonoids, (rosmarinic acid and eupatorin) with high antioxidant activity. Ethanolic (50 %) and methanolic extracts using soxhlet showed high proteins and glycosaponins. Water extracts using reflux and maceration showed high polysaccharides. Methanolic extract (50 %) using soxhlet and methanolic extract using maceration showed strong cytotoxic effect against MCF7 and HCT116 cell lines, respectively. Antioxidant and cytotoxic activities showed significant correlation with selected primary and secondary metabolites. HPLC fingerprints combined with chemometrics showed the extracts have been clustered based on selected major peaks profile. FTIR fingerprints combined with chemometrics showed that the extracts have been clustered based on protein and polysaccharide contents. Conclusion Ten different extracts of O. stamineus have showed significant differences in the content of selected primary and secondary metabolites as well as the biological activities. Chemometric tools were able to classify and discriminate the distinctive features of extracts thus can be correlated with the biological activities.

Most herbs have their own characteristic smell due to the presence of volatile compounds. Traditionally, volatiles are analyzed by using sophisticated and expensive gas chromatography (GC) in tandem with a selective mass spectrometric (MS) detector. An alternative approach is described based on the use of an electronic nose. The approach is much simpler than the traditional GC–MS counterparts, but providing key information of the samples analyzed. Using lipids and gas chromatography stationary phase materials with different polarities as sensing membranes, a quartz crystal microbalance smell sensor array has been developed for the analysis of traditional medicinal plants. The headspace vapors of different types of Tongkat Ali (Eurycoma longifolia) extracts were analyzed by the smell sensor and GC–MS. Correlation between the sensor response and the identified compounds were studied using principal component analysis. Some of the identified compounds exhibited good correlation with the quartz crystal microbalance (QCM) sensor array data.

This preliminary work investigates whether Fourier transform infrared spectroscopy (FT-IR), in combination with multivariate analysis can be used to distinguish Phyllanthus niruri Linn. samples from different locations. A FT-IR spectrometer used in this work is equipped with a transmittance accessory. All samples were scanned in the mid-infrared range of 400–4000 cm −1 but only the region of 400–2000 cm −1 that contained the most spectral information was analyzed with multivariate techniques of principal component analysis (PCA), genetic algorithm (GA), linear discriminant analysis (LDA) and SIMCA. PCA was used to reduce the spectra to three principal components, which explained 96.2% of the whole variance and a plot of principal components two and three showed groupings of the samples according to origin. Secondly, the spectra compressed with PCA were used to develop a LDA. The GA applied for 50 or 100 generations used concept of maximizing the ratio of between-groups variance to within-groups variance and identified the most discriminating variable for subsequent LDA. The SIMCA technique consisted of constructing an enclosure for each location using separate principal component models and using these models to classify the whole data. The best discriminatory approach was the technique using GA with LDA, where using just six wavenumber variables obtained from application of GA for 100 generations, gave 100% correct prediction for both originally grouped and cross-validated data. However, both PCA–LDA and SIMCA also gave very good classification. Basically, FT-IR analysis is rapid and has the power to discriminate samples in terms of geographical origin with the aid of multivariate analyses.

The modified Schneider's gel slide method is used to monitor the inhibitory effect of 50% methanol extract of Orthosiphon stamineus on the growth of calcium oxalate crystals. The images of crystals grown in the gel were captured at 1, 2, 3, 4 and 24 h with digital camera and image analysis was carried out. A total of nine variables relating to size and shape parameters were calculated and six were used for further analysis. Principal component analysis (PCA) and self-organizing map (SOM) were applied in the visualization of the size and shape distribution of the produced crystals using the following six variables: area, convex area, aspect ratio, equivalent diameter, roundness and full ratio. The results indicate that the modified gel slide method with the use of PCA and SOM to reduce the dimensionality of the data allowed an intuitive presentation of the differences in the studied inhibitors. The decrease in crystal growth for the extract and positive control of sodium citrate was clearly evident in the principal component scores and also on the location in the SOM. Evidence about the decrease in growth was also provided by an inhibition index of 0.290 for the extract and 0.255 for the positive control.

A PVC blended lipid membrane coated quartz crystal microbalance has been prepared to mimic human olfaction system. Transient response curves of the sensor on exposure to methanol, ethanol, chloroform, acetone and benzene were studied. Transient parameters, viz., simple parameters consisting of peak heights, derivatives, slopes and integrals, and polynomial parameters consisting of coefficients from the curve fitting equations, were extracted from the transient response curves and used as data for the subsequent chemometric data analysis. The sensor showed good separation and classification of VOCs.

Objective: To investigate the effect of methanolic extract of Melastoma malabathricum (M. malabathricum) in thrombocyte counts in mice. Methods: Methanolic extract of M. malabathricum corresponding to 1.5 to 2 mg/10 g body weight in saline was administered to mices via oral route. Control group was given normal saline. Twenty five microlitres of blood were drawn at 0 h and thereafter at 1, 2, 4, 8, 12, 20 h after dosing via tail bleeding technique. The thrombocytes were counted in the triple laminated middle 25 squares of the haemocytometer using light microscope. Results: The control group showed a moderate rise after the administration of saline whereas the M. malabathricum treated group shows significant rise within 2nd h in thrombocyte counts with an increment of 51.64% compared to baseline count. Conclusions: Based on the results, it can be concluded that M. malabathricum could be a potential remedy in treating thrombocytopenic condition.

Phytochemical investigation on the leaves of Blumea balsamifera DC resulted in the isolation of 11 flavonoids. Their chemical structures were elucidated by means of elemental analyses and different spectroscopic methods, such as UV, IR, NMR and MS. The free radical scavenging activity of organic extracts of B. balsamifera DC leaves and that of pure flavonoids isolated from the leaves was evaluated using 1,1-diphenyl-2-picrylhydrazyl radical. A dose response curve was plotted for determining SC50 values (the concentrations required to inhibit radical formation by 50%). The antioxidant activities of crude extracts decreased in the order: methanol extract > chloroform extract > pet-ether extract. The antioxidant activities of all compounds tested decreased in the order: quercetin > rhamnetin > luteolin > luteolin-7-methylether > l -ascorbic acid > blumeatin > butylated hydroxyanisole > 5,7,3′,5′-tetrahydroxyflavanone > tamarixetin > butylated hydroxytoluene > α-tocopherol > dihydroquercetin-4′-methylether > dihydroquercetin-7,4′-dimethylether. This result indicates that flavonoid contents of different solvent extracts of B. balsamifera DC leaves were responsible for their antioxidant properties.

With the proliferation of numerous herbal remedies on the market, the ability to verify the actual presence of the active herbal ingredient in a product would be very useful. In this paper, a microcontroller-based electronic taste sensing system capable of discriminating between liquid samples containing Eurycoma longifolia and those that do not, is described. The ‘taste’ of the liquid sample is sensed using specially fabricated disposable screen-printed array of non-specific lipid-membrane sensors and classified by means of artificial neural network (ANN). The overall system is controlled by an embedded microcontroller, which performed the data acquisition, the ANN-based pattern recognition and the user interface tasks. From the extensive tests that were performed, excellent recognition results have been obtained. The system is flexible and could easily be trained for other herbal samples.

This study evaluates the primary and secondary metabolite profiles of Eurycoma longifolia Jack (EL) stems and leaves to determine whether it can be utilized for therapeutic purposes as the roots. A total of six types of extracts were tested. The extracts showed high content of glycosaponins, polysaccharides, proteins and phenolics. The presence of flavonoids and phospholipids was also detected. High Performance Thin Layer Chromatography (HPTLC) and High Performance Liquid Chromatography (HPLC) analysis showed the presence of bioactive marker of EL root, eurycomanone and 14,15AŸ-dihydroxyklaineanone in stem and leaf extracts. Primary and secondary metabolites identified were reported to associate with the enhancement of ergogenic and aphrodisiac activities in animal and human subjects. The result shows that stem and leaves of E. longifolia has the potential for therapeutic purposes.

Xanthones are a group of oxygenated heterocyclic compounds with anticancer properties, but poor aqueous solubility and low oral bioavailability hinder their therapeutic application. This study sought to prepare a xanthones extract (81% α-mangostin and 16% γ-mangostin) in polymeric nanoparticles and to investigate its intracellular delivery and cytotoxicity toward colon cancer cells. The nanoparticles were prepared in Eudragit RL100 and Eudragit RS100 by the nanoprecipitation method at drug loading and entrapment efficiency of 20% and >95%, respectively. Freeze-drying of bulk nanoparticle solutions, using glucose or sucrose as cryoprotectants, allowed the collection of nanoparticles at >95% yield. Solubility of the xanthones extract was improved from 0.1 µg/mL to 1250 µg/mL. Transmission electron microscopy (TEM) and dynamic light scattering (DLS) of the freeze-dried final formulation showed the presence of cationic round nanoparticles, with particle size in the range of 32–130 nm. Scanning electron microscopy (SEM) showed the presence of nanospheres, and Fourier transform infrared (FTIR) spectroscopy indicated intermolecular interaction of xanthones with Eudragit polymers. Cellular uptake of nanoparticles was mediated via endocytosis and indicated intracellular delivery of xanthones associated with potent cytotoxicity (median inhibitory concentration26.3±0.22 µg/mL). Presented results suggest that cationic nanoparticles of xanthones may provide a novel oral drug delivery system for chemoprevention or treatment of intestinal and colon tumors.

Obesity is one of the major public health problems worldwide and it is generally associated with many diseases. Although synthetic drugs are available for the treatment of obesity, herbal remedies may provide safe, natural, and cost-effective alternative to synthetic drugs. One example of such drugs is Melastoma malabathricum var Alba Linn (MM). Although several studies have been reported for the pharmacological activities of MM, there is no report on the anti-obesity effect of MM. The aim of the present study is to evaluate the anti-obesity potential of methanolic extract of MM. The anti-obesity effect of MM on rats fed with a high-fat diet was investigated through determination of the changes in body weight, fat weight, organ weights, and blood biochemicals. The animals in this study were divided into three groups: a normal group with a standard diet (N), a control group fed with high-fat diet (C), and a MM treatment group fed with high-fat (HFD + MM) diet for 8 weeks. There was no significant difference in the amount of food intake between control and HFD + MM treatments. These results also suggest that MM does not induce a dislike for the diet due to its smell or taste. The study shows that MM significantly prevented increases in body weight, cholesterol, LDL, HDL, and total lipids that resulted from the high-fat diet. MM also decreased the epididymal fat (E-fat) and retroperitoneal fat (R-fat) weights and phospholipid concentrations induced by the high-fat diet. On the basis of these findings, it was concluded that MM had anti-obesity effects by suppressing body weight gain and abdominal fat formation.

Tephrosia apollinea is a perennial shrublet widely distributed in Africa and is known to have medicinal properties. The current study describes the bio-assay (cytotoxicity) guided isolation of (-)-pseudosemiglabrin from the aerial parts of T. apollinea. The structural and stereochemical features have been described using spectral and x-ray crystallographic techniques. The cytotoxicity of isolated compound was evaluated against nine cancer cell lines. In addition, human fibroblast was used as a model cell line for normal cells. The results showed that (-)-pseudosemiglabrin exhibited dose-dependent antiproliferative effect on most of the tested cancer cell lines. Selectively, the compound showed significant inhibitory effect on the proliferation of leukemia, prostate and breast cancer cell lines. Further studies revealed that, the compound exhibited proapoptotic phenomenon of cytotoxicity. Interestingly, the compound did not display toxicity against the normal human fibroblast. It can be concluded that (-)-pseudosemiglabrin is worthy for further investigation as a potential chemotherapeutic agent.

Recently, there has been increasing interest in Ficus deltoidea (Moracea) due to its chemical composition and the potential health benefits. The leaves of the plant have been suggested to have potential antidiabetic effects. Inhibition of carbohydrate-hydrolysing enzymes, such as α-glucosidase and α-amylase is one of the therapeutic approaches to control postprandial hyperglycemia. In this study, enzymes inhibitory effect and antioxidant properties of different fractions of methanolic extract obtained from F. deltoidea leaves was evaluated. Further, the possible relationship between pharmacological properties and phytochemical content of fractions was investigated. The n-butanol fraction showed significant α-glucosidase and α-amylase inhibitory effects (IC 50 values 15.1 and 39.42 μg/ml, respectively) along with the remarkable antioxidant activity when compared to the other fractions. High performance liquid chromatography (HPLC) chemical profiling of the n-butanol fraction revealed that the contents of isovitexin (24.63 mg/g) and vitexin (8.3 mg/g) were found to be significantly higher than the other fractions. These results indicate that F. deltoidea could be the potential source of promising anti-diabetic drug. Key words : Ficus deltoidea, enzymes, phytochemical, high performance liquid chromatography (HPLC), isovitexin, vitexin.

Background Syzygium campanulatum Korth (Myrtaceae) is an evergreen shrub rich in phenolics, flavonoid antioxidants, and betulinic acid. This study sought to investigate antiangiogenic and anti-colon cancer effects of S.C. standardized methanolic extract. Methods Betulinic acid was isolated from methanolic extract by crystallization and chromatography techniques. S.C. methanolic extract was analyzed by UV-Vis spectrophotometry, FTIR, LC-MS, and HPLC. Antiangiogenic effect was studied on rat aortic rings, matrigel tube formation, cell proliferation and migration, and expression of vascular endothelial growth factor (VEGF). Antitumor effect was studied using a subcutaneous tumor model of HCT 116 colorectal carcinoma cells established in nude mice. Results Analysis by HPLC, LC-MS and FTIR confirm presence of betulinic acid in S.C. methanolic extract. Quantitative analysis by HPLC indicates presence of betulinic acid in S.C. extract at 5.42 ± 0.09% (w/w). Antiangiogenesis study showed potent inhibition of microvessels outgrowth in rat aortic rings, and studies on normal and cancer cells did not show any significant cytotoxic effect. Antiangiogenic effect was further confirmed by inhibition of tube formation on matrigel matrix that involves human endothelial cells (IC50 = 17.6 ± 2.9 μg/ml). S.C. extract also inhibited migration of endothelial cells and suppressed expression of VEGF. In vivo antiangiogenic study showed inhibition of new blood vessels in chicken embryo chorioallantoic membrane (CAM), and in vivo antitumor study showed significant inhibition of tumor growth due to reduction of intratumor blood vessels and induction of cell death. Conclusion Collectively, our results indicate S. campanulatum as antiangiogenic and antitumor candidate, and a new source of betulinic acid.

OBJECTIVE: Ficus deltoidea leaves have been used in traditional medicine in Southeast Asia to treat diabetes, inflammation, diarrhea, and infections. The present study was conducted to assess the genotoxicity and acute and subchronic toxicity of a standardized methanol extract of F. deltoidea leaves. METHODS: Sprague Dawley rats were orally treated with five different single doses of the extract and screened for signs of toxicity for two weeks after administration. In the subchronic study, three different doses of the extract were administered for 28 days. Mortality, clinical signs, body weight changes, hematological and biochemical parameters, gross findings, organ weights, and histological parameters were monitored during the study. Genotoxicity was assessed using the Ames test with the TA98 and TA100 Salmonella typhimurium strains. Phytochemical standardization was performed using a colorimeter and high-performance liquid chromatography. Heavy metal detection was performed using an atomic absorption spectrometer. RESULTS: The acute toxicity study showed that the LD50 of the extract was greater than 5000 mg/kg. In the subchronic toxicity study, there were no significant adverse effects on food consumption, body weight, organ weights, mortality, clinical chemistry, hematology, gross pathology, or histopathology. However, a dose-dependent increase in the serum urea level was observed. The Ames test revealed that the extract did not have any potential to induce gene mutations in S. typhimurium, either in the presence or absence of S9 activation. Phytochemical analysis of the extract revealed high contents of phenolics, flavonoids, and tannins. High-performance liquid chromatography analysis revealed high levels of vitexin and isovitexin in the extract, and the levels of heavy metals were below the toxic levels. CONCLUSION: The no-observed adverse effect level of F. deltoidea in rats was determined to be 2500 mg/kg.

The leaves of Blumea balsamifera are used as a folk medicine in kidney stone diseases in South-East Asia. Phytochemical investigation revealed leaves contained a number of flavonoids. In view of these, the present work was aimed to quantify and preliminary pharmacokinetic investigation of five flavonoids viz. dihydroquercetin-7,4¢-dimethyl ether (I), dihydroquercetin-4¢-methyl ether (II), 5,7,3¢,5¢-tetrahydroxyflavanone (III), blumeatin (IV) and quercetin (V) in rat plasma following oral administration (0.5g/Kg) of B. balsamifera leaf extract in rats. Quantification was achieved by using a validated, reproducible high-performance liquid chromatographic method. The mean recoveries of I, II, III, IV and V were 90.6, 93.4, 93.5, 91.2 and 90.3% respectively. The limit of quantification was 25 ng/mL for I and IV, 10 ng/mL for II and III and 100 ng/mL for V respectively. The within day and day-to-day precision for all the compounds were10%. The validated HPLC method herein was applied for pharmacokinetic studies and the main pharmacokinetic parameters were: t1/2 (hr) 5.8, 4.3, 2.9, 5.7 and 7.3, Cmax (ng/mL) 594.9, 1542.9 1659.9, 208.9 and 3040.4; Tmax (hr) 4.7, 1.0, 1.0, 3.5 and 2.3; AUC0-oo (ng hr/mL) 5040, 5893, 9260, 1064 and 27233 for I, II, III, IV and V respectively. The developed method was suitable for pharmacokinetic studies and this preliminary study also revealed significant absorption after oral dosing in rats.

Background: Orthosiphon stamineus (OS) Benth is a medicinal plant and native in Southeast Asia. Previous studies have shown that OS leaves possess antioxidant, cytotoxic, diuretic, antihypertensive, and uricosuric effects. These beneficial effects have been attributed to the presence of primary and secondary metabolites such as polyphenols, amino acids, and flavonoids. Objective: To develop and validate an high-performance liquid chromatography (HPLC)-diode array detector (DAD) method combined with solid-phase extraction that involves precolumn derivatization with O-phthaladehyde for simultaneous analysis of free amino acids in OS leaves extracts. Materials and Methods: OS leaves were extracted with water (OS-W), ethanol (OS-E), methanol (OS-M), 50% ethanol (OS-EW), and 50% methanol (OS-MW). The extracts were treated by C18 cartridge before derivatization, resulting in great improvement of separation by Zorbox Eclipse XDB-C18 column. Results: The HPLC–DAD method was successfully developed and validated for analyzing the contents of free amino acids in OS extracts. The results showed that l-aspartic acid with 0.93 ± 0.01 nmol/mg was the major free amino acid in OS-W extract. However, in OS-E, OS-M, OS-EW, and OS-MW, l-glutamic acid with 3.53 ± 0.16, 2.17 ± 0.10, 4.01 ± 0.12, and 2.49 ± 0.12 nmol/mg, respectively, was the major free amino acid. Subsequently, l-serine, which was detected in OS-W, OS-E, and OS-M, was the minor free amino acid with 0.33 ± 0.02, 0.12 ± 0.01, and 0.06 ± 0.01 nmol/mg, respectively. However, l-threonine with 0.26 ± 0.02 and 0.19 ± 0.08 nmol/mL in OS-EW and OS-MW, respectively, had the lowest concentration compared with other amino acid components. Conclusion: All validation parameters of the developed method indicate that the method is reliable and efficient to simultaneously determine the free amino acids content for routine analysis of OS extracts. Abbreviations used: HPLC-DAD: High-Performance Liquid Chromatography with Diode-Array Detection, OS: Orthosiphon stamineus, OS-W: Orthosiphon stamineus water extract, OS-E: Orthosiphon stamineus ethanol extract, OS-M: Orthosiphon stamineus methanol extract, OS-EW: Orthosiphon stamineus 50% ethanol extract, OS-MW: Orthosiphon stamineus 50% methanol extract, OPA: O-phthaladehyde, SPE: Solid Phase Extraction, UV: Ultraviolet, LOD: Limit of Detection, LOQ: Limit of Quantification, RSD: Relative Standard Deviation.

Objective: This study aimed to perform pharmacokinetic profile of rosmarinic acid (RA), sinensitin (SIN), eupatorin (EUP) and 3΄-hydroxy-5,6,7,4΄-tetramethoxyflavone (TMF) in Orthosiphon stamineus ethanolic extract (OS-E) and its nanoliposomes (OS-EL) after oral and intravenous administration in Sprague-Dawley rat’s plasma by developing and validating a high-performance liquid chromatography (HPLC) method with ultraviolet (UV) detection.Methods: An isocratic elution program consisting of methanol: tetrahydrofuran: water (0.1% H3PO4) mixture in the volume ratio 55: 5: 40 on Nucleosil C18 column (250 × 4.6 mm internal diameter × 5 µm particles size) was applied. The current study followed a two-ways crossover study design. OS-E and OS-EL were administered orally at 1000 and 500 mg/kg, respectively. They were also administered intravenously at 250 mg/kg via the tail vein.Results: The HPLC-UV method was successfully developed and validated for simultaneous determination of major chemical constituent from OS-E and OS-EL in rat’s plasma. The method recorded the mean recoveries from extraction were between 91.39 and 100.32%. With regards to the intravenous administration of OS-EL, all four marker compounds appeared to be poorly distributed and cleared slowly from the body compared to OS-E. Whilst in oral administration of OS-EL, the bioavailability of all marker compounds were higher than OS-E due to higher solubility of encapsulation in phospholipids.Conclusion: The higher solubility and bioavailability of OS-EL may contribute to encapsulation in phospholipids.

Purpose: To develop a rapid and reliable reverse phase high performance liquid chromatography (RPHPLC) method to quantify the two isomers of asarones in P. sarmentosum extracts using two different columns under similar analytical conditions. Methods: Two isomers, α- and β-asarone, were analyzed using two types of C 18 columns with 0.1 % orthophosphoric acid: acetonitrile: methanol (50: 40: 10) as mobile phase. The developed method was applied to determine the contents of α- and β-asarone in extracts of different parts of P. sarmentosum. Results: Column A retention times for the elution of α- and β-asarone were 11.890 ± 0.008 and 10.80 ± 0.004 min, respectively, and were significantly shorter than those of column B (15.110 ± 0.024 and 13.290 ± 0.018, respectively, p < 0.001). Column B showed better resolution (1.82 ± 0.025 of the isomers than column A (1.10 ± 0.01, p < 0.001). Both columns showed comparable sensitivity, precision and selectivity of the compounds investigated. α-Asarone level was in the range 0.36 - 5.14 % in ethanol and 50 % ethanol extracts, but absent in all water extracts. β-Asarone occurred in the range of 0.01 - 0.15 % in ethanol and 50 % ethanol extracts but was absent in all water extracts of P. sarmentosum . Conclusion: The results indicate that the developed method is a suitable quality assurance method for determining α- and β-asarone isomers in herbal extracts and food preparations. Keywords: α- Asarone, Isomers, Piper sarmentosum , Herbal extracts, Retention time

Garcinia mangostana fruit rinds contain high concentration of xanthones such as α-, and γ-mangostin. The α-mangostin-rich extracts have been used widely in nutritional supplements, herbal cosmetics and pharmaceutical preparations. This study aims to develop a reverse phase HPLC method for the quantification of α-, β- and γ-mangostin in G. mangostana fruit rind extracts. The method was validated at 244, 254, 316 and 320 nm. Selectivity was determined by comparing the retention time and the UV-Vis spectra of α-, β- and γ-mangostin in G. mangostana extracts with those of the reference compounds. Linearity was in the range 0.2 to 200 µg/ml at R2 > 0.9999. The intraday and interday precision was determined as a relative standard deviation, and was found to be (0.4±0.4) and (0.3 ± 0.3)%, respectively. The percentage recovery was in the range (96.3 ± 2.5) to (100.5 ± 3.4)%. The limits of detection and quantification were in the range 0.06 to 0.12 and 0.14 to 0.37 µg/ml, respectively. The reported method was applied for the determination of α-, β- and γ-mangostin concentration in 7 extracts of G. mangostana fruit rinds, and it could be considered as an important analytical tool for quality control, stability studies, pharmacokinetics, and standardization purposes. Key words: Alpha-, beta-, and gamma-mangostin, Garcinia mangostana, mangosteen.

Background Xanthones are a group of oxygen-containing heterocyclic compounds with remarkable pharmacological effects such as anti-cancer, antioxidant, anti-inflammatory, and antimicrobial activities. Methods A xanthones extract (81% α-mangostin and 16% γ-mangostin), was prepared by crystallization of a toluene extract of G. mangostana fruit rinds and was analyzed by LC-MS. Anti-colon cancer effect was investigated on HCT 116 human colorectal carcinoma cells including cytotoxicity, apoptosis, anti-tumorigenicity, and effect on cell signalling pathways. The in vivo anti-colon cancer activity was also investigated on subcutaneous tumors established in nude mice. Results The extract showed potent cytotoxicity (median inhibitory concentration 6.5 ± 1.0 μg/ml), due to induction of the mitochondrial pathway of apoptosis. Three key steps in tumor metastasis including the cell migration, cell invasion and clonogenicity, were also inhibited. The extract and α-mangostin up-regulate the MAPK/ERK, c-Myc/Max, and p53 cell signalling pathways. The xanthones extract, when fed to nude mice, caused significant growth inhibition of the subcutaneous tumor of HCT 116 colorectal carcinoma cells. Conclusions Our data suggest new mechanisms of action of α-mangostin and the G. mangostana xanthones, and suggest the xanthones extract of as a potential anti-colon cancer candidate.

Background Xanthones are a group of oxygen-containing heterocyclic compounds with remarkable pharmacological effects such as anti-cancer, antioxidant, anti-inflammatory, and antimicrobial activities. Methods A xanthones extract (81% α-mangostin and 16% γ-mangostin), was prepared by crystallization of a toluene extract of G. mangostana fruit rinds and was analyzed by LC-MS. Anti-colon cancer effect was investigated on HCT 116 human colorectal carcinoma cells including cytotoxicity, apoptosis, anti-tumorigenicity, and effect on cell signalling pathways. The in vivo anti-colon cancer activity was also investigated on subcutaneous tumors established in nude mice. Results The extract showed potent cytotoxicity (median inhibitory concentration 6.5 ± 1.0 μg/ml), due to induction of the mitochondrial pathway of apoptosis. Three key steps in tumor metastasis including the cell migration, cell invasion and clonogenicity, were also inhibited. The extract and α-mangostin up-regulate the MAPK/ERK, c-Myc/Max, and p53 cell signalling pathways. The xanthones extract, when fed to nude mice, caused significant growth inhibition of the subcutaneous tumor of HCT 116 colorectal carcinoma cells. Conclusions Our data suggest new mechanisms of action of α-mangostin and the G. mangostana xanthones, and suggest the xanthones extract of as a potential anti-colon cancer candidate.

Piper sarmentosum Roxb. (Piperaceae) is a traditional medicinal as well as a culinary plant in South East Asian countries, whereby aerial parts of the plant are consumed as a vegetable in various forms and the whole plant or parts are used as folk remedies, alone or in combination with other herbs, to treat various ailments. The plant has extensively been investigated in a broad range of studies to provide scientific evidence for folklore claims or to find new therapeutic uses; however, heretofore, a summary of the data are not available.In order to describe nutritional and therapeutic potential of P. sarmentosum and summarize scientific evidence that supports traditional claims, a literature review and latest advances in research of the plant are given herein.The literature has been retrieved from a number of databases such as Google Scholar, PubMed, Medline, Science Direct and SciFinder. The articles related to synthetic work, ecology and agriculture have been excluded.The review has not only revealed a number of pharmacological activities supporting the traditional claims but indicates new prospects for the plant. Antiangiogenic activity and toxicity studies suggest the usage of the plant in treating diseases involving neo-vascularization. The available efficacy, safety, pharmacokinetic and stability data urge clinical studies on extracts of the plant.The present review may be helpful to future researchers intending to investigate the plant and natural pharmaceutical industry for preparing evidence-based formulations.

Hitherto, only a few studies are reported about using the combination of TLC and RP-HPLC for the separation and determination of analyte(s) from a complex matrix. The present study is aimed to develop a simple and rapid method for the separation and determination of betulinic acid from a complex matrix, extracts of Orthosiphon stamineus, using a combination of the two techniques. The samples having higher contents of the analyte and fewer interfering species were prepared using TLC. The samples were then eluted through C(18) column using isocratic solvent system comprising acetonitrile, methanol and acetic acid acidified water of pH 2.8 in a ratio of 70 : 20 : 10 (v/v/v), respectively, and detection was carried out at 210 nm. The method was validated and applied successfully to quantify betulinic acid in various types of extracts of the plant. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.0005 and 0.0050 μg/ml, respectively. The method exhibited linearity in a concentration range of 0.005-100.00 μg/ml (R(2)= 0.9999). The recovery was found to be 97.10 - 97.60% (RSD5%), whereas, intra-day and inter-days accuracy values were 97.13 - 98.67% (RSD5%) and 96.45 - 98.00% (RSD5%), respectively. The results of the present study indicate that the developed method is simple, rapid, sensitive and accurate, and may be of a value to natural product industry and researchers for the standardization of extracts containing betulinic acid in a lesser time and consuming fewer solvents.

The Clerodendrum serratum (Spreng.) known as ‘Timba Tasek’ is widely used in Asian countries especially Malaysia as the traditional medicine to treat various diseases. This study aimed to evaluate the antioxidant, antiangiogenic and vasorelaxant activities as well as the chemical profiles of C. serratum leaves extract. The dried powder leaves of C. serratum were extracted serially with petroleum ether, chloroform, followed by methanol and water by maceration method. To elucidate the antiangiogenic properties, the inhibitory effects of these extracts on blood vessel growth formation were adapted in rat aortic ring assay. In another set of experiments, the possible vasorelaxant activity of C. serratum leaves extracts were examined on an isolated rat aortic ring preparations and responses of cumulative doses of noradrenaline (NA) were used. To determine antioxidant activity of this plant, the present study used well-established methods, that is, 2,2’-diphenyl-1-picrylhydrazyl (DPPH•) radical scavenging activity and trolox equivalent antioxidant capacity (TEAC) assay. The results showed that, amongst four extracts, methanolic extract of C. serratum (ME-CS) showed the most potent antioxidant, antiangiogenic and vasorelaxant activities. In another hand, qualitative study proved that ME-CS contains polyphenolics (hydrolysable tannins and flavonoids), terpenoids, saponins and may not contain any alkaloids. Therefore, while polyphenolics are the predominant compounds found in ME-CS, it is highly probable that they may play an important (dominant) role in antioxidant, antiangiogenic and vasorelaxant activity. Since all the three activities of C. serratum extracts end up in the same results, it is likely that, all the activities were contributed by same group (such as polyphenolics) or totally different group of chemical compounds that may act synergistically together with polyphenolics. Polyphenolics are responsible for antioxidant, antiangiogenic and vasorelaxant effects of plants as herbal therapy such as C. serratum leaves. Key words: Clerodendrum serratum (Spreng.) leaves, polyphenolics, antioxidant, antiangiogenic, vasorelaxant, phytochemical analysis.

Parkia speciosa Hassk is a traditional medicinal plant with strong antioxidant and hypoglycemic properties. This study aims to investigate the total phenolic content, antioxidant, cytotoxic and antiangiogenic effect of eight extracts from P. speciosa empty pods. The extracts were found to contain high levels of total phenols and demonstrated strong antioxidant effect in DPPH scavenging test. In rat aortic rings, P. speciosa extracts significantly inhibited the microvessel outgrowth from aortic tissue explants by more than 50%. The antiangiogenic activity was further confirmed by tube formation on matrigel matrix involving human endothelial cells. Cytotoxic effect was evaluated by XTT test on endothelial cells as a model of angiogenesis versus a panel of human cancer and normal cell lines. Basically the extracts did not show acute cytotoxicity. Morphology examination of endothelial cells indicated induction of autophagy characterized by formation of plenty of cytoplasmic vacuoles. The extracts were found to work by decreasing expression of vascular endothelial growth factor in endothelial cells.

Cat's whiskers (Orthosiphon stamineus) is commonly used as Java tea to treat kidney stones including a variety of angiogenesis-dependent diseases such as tumorous edema, rheumatism, diabetic blindness, and obesity. In the present study, antitumor potential of standardized 50% ethanol extract of O. stamineus leaves (EOS) was evaluated against colorectal tumor in athymic mice and antiangiogenic efficacy of EOS was investigated in human umbilical vein endothelial cells (HUVEC). EOS at 100 mg/kg caused 47.62 ± 6.4% suppression in tumor growth, while at 200 mg/kg it caused 83.39 ± 4.1% tumor regression. Tumor histology revealed significant reduction in extent of vascularization. Enzyme-linked immunosorbent assay showed EOS (200 mg/kg) significantly reduced the vascular endothelial growth factor (VEGF) level in vitro (211 ± 0.26 pg/ml cell lysate) as well as in vivo (90.9 ± 2 pg/g tissue homogenate) when compared to the control (378 ± 5 and 135.5 ± 4 pg, respectively). However, EOS was found to be noncytotoxic to colon cancer and endothelial cells. In vitro, EOS significantly inhibited the migration and tube formation of human umbilical vein endothelial cells (HUVECs). EOS suppressed VEGF-induced phosphorylation of VEGF receptor-2 in HUVECs. High performance liquid chromatography (HPLC) analysis of EOS showed high rosmarinic acid contents, whereas phytochemical analysis revealed high protein and phenolic contents. These results demonstrated that the antitumor activity of EOS may be due to its VEGF-targeted antiangiogenicity.

Deregulated cell signaling pathways result in cancer development. More than one signal transduction pathway is involved in colorectal cancer pathogenesis and progression. Koetjapic acid (KA) is a naturally occurring seco-A-ring oleanene triterpene isolated from the Sandoricum koetjape stem bark. We report the cellular and molecular mechanisms of anticancer activity of KA towards human colorectal cancer. The results showed that KA induces apoptosis in HCT 116 colorectal carcinoma cells by inducing the activation of extrinsic and intrinsic caspases. We confirmed that KA-induced apoptosis was mediated by DNA fragmentation, nuclear condensation and disruption in the mitochondrial membrane potential. Further studies on the effect of KA on cancer pathways show that the compound causes down-regulation of Wnt, HIF-1α, MAP/ERK/JNK and Myc/Max signaling pathways and up-regulates the NF-κB signaling pathway. The result of this study highlights the anticancer potential of KA against colorectal cancer.

Background Angiogenesis, the formation of new blood vessels, has become an important target in cancer therapy. Angiogenesis plays an important role in tumor growth and metastasis. Koetjapic acid (KA) is a seco-A-ring oleanene triterpene isolated from S. koetjape. The solvent extract of this plant species was shown previously to have strong antiangiogenic activity; however the active ingredient(s) that conferred the biological activity and the mode of action was not established. Given the high concentration of KA in S. koetjape, an attempt has been made in this study to investigate the antiangiogenic properties of KA. Results Treatment with 10-50 μg/ml KA resulted in dose dependent inhibition of new blood vessels growth in ex vivo rat aortic ring assay. KA was found to be non-cytotoxic against HUVECs with IC50 40.97 ± 0.37 μg/ml. KA inhibited major angiogenesis process steps, endothelial cell migration and differentiation as well as VEGF expression. Conclusions The non-cytotoxic compound, KA, may be a potent antiangiogenic agent; its activity may be attributed to inhibition of endothelial cells migration and differentiation as well VEGF suppression.

In vitroassays are economical and easy to perform but to establish relevance of their results to real clinical outcome in animals or human, pharmacokinetics is prerequisite. Despite variousin vitropharmacological activities of extracts ofPiper sarmentosum, there is no report of pharmacokinetics. Therefore, the present study aimed to evaluate ethanol extract of fruit of the plant in dose of 500 mg kg−1orally for pharmacokinetics. Sprague-Dawley rats were randomly divided into groups 1, 2, and 3 (eachn= 6) to study absorption, distribution and excretion, respectively. High performance liquid chromatography (HPLC) with ultraviolet detection was applied to quantify pellitorine, sarmentine and sarmentosine in plasma, tissues, feces and urine to calculate pharmacokinetic parameters. Pellitorine exhibited maximum plasma concentration (Cmax) 34.77 ng mL−1± 1.040, time to achieveCmax(Tmax) 8 h, mean resident time (MRT) 26.00 ± 0.149 h and half life (t1/2) 18.64 ± 1.65 h. Sarmentine showedCmax191.50 ± 12.69 ng mL−1,Tmax6 h, MRT 11.12 ± 0.44 h andt1/210.30 ± 1.98 h. Sarmentosine exhibited zero oral bioavailability because it was neither detected in plasma nor in tissues, and in urine. Pellitorine was found to be distributed in intestinal wall, liver, lungs, kidney, and heart, whereas sarmentine was found only in intestinal wall and heart. The cumulative excretion of pellitorine, sarmentine and sarmentosine in feces in 72 h was 0.0773, 0.976, and 0.438 μg, respectively. This study shows that pellitorine and sarmentine have good oral bioavailability while sarmentosine is not absorbed from the gastrointestinal tract.

Blumea balsamifera DC (Compositae) leaves have been recommended for use as a folk medicine in the treatment of various diseases related to urolithiasis in southeast Asia. Phytochemical studies of this plant revealed it contains four classes of flavonoids (e.g., flavonols, flavones, flavanones, and dihydroflavonol derivatives).In view of the broad pharmacological activity of flavonoids, this study was carried out to determine the xanthine oxidase (XO) inhibitory and enzymatically produced superoxide radical scavenging activity of different organic extracts and that of the isolated flavonoids from B. balsamifera leaves.The inhibitory activity of XO was assayed spectrophotometrically at 295 nm. The superoxide radicals scavenging activity was assessed by NBT reduction method, spectrophotometrically at 560 nm. A dose response curve was plotted for determining IC₅₀ values.The methanol extract (IC₅₀ = 0.111 mg/mL) showed higher XO inhibitory activity than the chloroform (0.138 mg/mL) and pet-ether extracts (0.516 mg/mL). IC₅₀ values of scavenging of superoxide radicals for extracts decreased in the order of: methanol (0.063 mg/mL) chloroform (0.092 mg/mL) pet-ether (0.321 mg/mL). The XO inhibitory activity of the isolated flavonoids and reference compounds tested decreased in the order of: allopurinol luteolin quercetin tamarixetin 5,7,3',5'-tetrahydroxyflavanone rhamnetin luteolin-7-methyl ether blumeatin dihydroquercetin-4'-methyl ether dihydroquercetin-7,4'-dimethyl ether L-ascorbic acid.The results indicated that the flavone derivatives were more active than the flavonol derivatives. The flavanone derivatives were moderately active and the dihydroflavonol derivatives were the least. The higher flavonoid content of extracts contributed to their higher XO inhibitory activity.

In the present study, L. ferrugineus methanol extract (LFME) was evaluated for its blood pressure lowering effect in anesthetized normotensive Sprague Dawley (SD) rats and its spasmogenic effect in isolated guinea pig ileum. The possible mechanism(s) of action were also investigated. LFME was obtained by Soxhlet extraction. The rats were fasted overnight and anesthetized with sodium pentobarbitone (60 mg/kg i.p.). LFME was administered in i.v. boluses in the concentrations of 25, 50, 100 and 200 mg/kg respectively, with concomitant monitoring of mean arterial pressure (MAP). It was found that LFME dose-dependently reduced MAP. An i.v. bolus injection of atropine significantly decreased the blood pressure lowering effect of LFME. Similarly, L-NAME (Nω-nitro-L-arginine methyl ester) significantly lowered both the MAP and the action duration. Conversely, no significant change in MAP was seen following i.v. injections of neostigmine, hexamethonium, prazosin and propranolol. LFME also produced a dose-dependent contractile effect in guinea pig ileum. This contraction was significantly reduced in atropine pre-incubated tissue segments, yet it was significantly enhanced in the presence of neostigmine. No appreciable change in the ability of LFME to contract guinea pig ileum was seen in the presence of hexamethonium. Accordingly, it can be postulated that LFME possesses a marked hypotensive effect that can be attributed to stimulation of muscarinic receptors and/or stimulation of nitric oxide (NO) release. Moreover, LFME retains a considerable spasmogenic action due to its cholinergic properties. The hypotensive and spasmogenic effects of LFME justify its traditional uses.

The present study aimed to investigate standardized ethanol extracts of fruit and leaves of Piper sarmentosum for their in vivo antioxidant activity in rats using a CCl (4)-induced oxidative stress model. The standardization was based on the quantification of the markers pellitorine, sarmentine and sarmentosine by high performance liquid chromatography (HPLC), and determination of total primary and secondary metabolites. The rats, divided into 7 groups each (n = 6), were used as follows: group 1 (CCl (4), negative control), group 2 (untreated, control), groups 3 and 4 (fruit extract 250 and 500 mg/kg, respectively), groups 5 and 6 (leaf extract 250 and 500 mg/kg, respectively) and group 7 (vitamin-E 100 mg/kg, positive control). The doses were administered orally for 14 days; 4 h following the last dose, a single dose of CCl (4) (1.5 mg/kg) was given orally to all the groups except group 2, and after 24 h, blood and liver of each animal were obtained. Analysis of plasma and liver homogenate exhibited significant preservation of markers of antioxidant activity, total plasma antioxidant activity (TPAA), total protein (TP), superoxide dismutase (SOD), catalase (CAT), and thiobarbituric acid reactive species (TBARS), in the pretreated groups as compared to the CCl (4) group (p < 0.05). Histology of the liver also evidenced the protection of hepatocytes against CCl (4) metabolites in the pretreated groups. The results of this study indicate the IN VIVO antioxidant activity of both extracts of the plant, which may be valuable to combat diseases involving free radicals.