Effects of Strobzlanthes Crispus Crude and Tea Extracts in Streptozotocin-Induced Hyperglycemic Rats

Strobilanthes crispus leaf has been used ethnomedically to treat diabetes mellitus and related disorders in Asia. The first part of this study is to develop a tea from leaves of S. crisps and investigate its antioxidant properties in vitro. Fermented and unfermented teas from young and old leaves of S. crispus were developed according to Camellia sinensis and Camellia theifera preparations for black and green tea, respectively. Three methods were used to determine the antioxidant activities i.e 1) p-carotene bleaching method 2) DPPH fiee radical scavenging assay 3) Ferric reducinglantioxidant power (FRAP) assay. The total phenolic content was also estimated using Folin-Ciocalteu method. The result showed that unfermented S. crisps tea displayed a higher antioxidant activity compared to fermented S. crispus tea. Tea developed from old or matured leaves possessed higher antioxidant activity compared to young leaves. However, commercial green (Sencha, UK) and black (Boh, Malaysia) tea that were developed from leaves of C. sinensis exhibited higher antioxidant activity among all teas tested. The second part of this study was aimed at determining the effect of S. crispus crude extract on STZ-induced hyperglycemic rats. S. crispus (young and old leaves) were extracted with distilled water and given to normal and hyperglycernic rats at concentrations of 2.5, 5.0 and 7.5% for 21 days. Plasma glucose, lipid profile (total cholesterol, triglyceride, HDL-cholesterol, LDL-cholesterol), total antioxidant status and serum potassium and magnesium contents were determined on baseline (day O), day 7 and day 21. The results showed that S. crispus crude extract at concentrations of 2.5, 5.0 and 7.5% from old or matured leaves reduced glucose level significantly in hyperglycemic rats (~4.05). Third part of this study evaluated the effect of S. crispus fermented and unfermented tea in STZ-induced hyperglycemic rats at a concentration of 2% for 21 days. Plasma glucose, lipid profile (total cholesterol, triglyceride, HDLcholesterol, LDL-cholesterol), total antioxidant status and serum potassium and magnesium contents were determined during baseline (day O), day 7 and day 21. The results showed that both fermented and unfermented S. crispus tea reduced glucose level in hyperglycemic rats (p<0.05). Fermented and unfermented S. crispus tea also improved antioxidant status and lipid profile in hyperglycemic rats by lowering the total cholesterol, triglyceride, and LDL-cholesterol. Total antioxidant status and HDLcholesterol also increased in hyperglycemic rats treated with fermented or unfermented tea S. crispus. Both fermented or unfermented S. crisps tea failed to prevent the reduction of serum magnesium in hyperglycemic rats

PHYSICOCHEMICAL, PHYTOCHEMICAL AND ANTIMICROBIAL PROPERTIES OF WILD HONEY COLLECTED AT MANGROVE AND MOUNTAIN AREAS IN SABAH, MALAYSIAN BORNEO

Objective: The aim of this study was to determine the physicochemical, phytochemical content and antimicrobial properties of selected honey of Sabah, Malaysian Borneo. Methods: A standardized protocols were used to evaluate the physicochemical properties of selected honey of Sabah while the phytochemicals content (phenolics and flavonoids) were determined using Folin-Ciocalteau and aluminium colorimetric methods. Antimicrobial properties were evaluated using disc diffusion assay. Results: For 80% methanol extract, old Upper Mountain honey contained the highest free acidity, conductivity, total phenolic and flavonoid contents with the values 23.84 ± 0.42 ml/g, 0.61 ± 0.01 mS/cm, 9.71 ± 0.01 mg gallic acid equivalent (GAE)/g and 7.76 ± 0.02 mg rutin equivalent (RU)/g, respectively. Antimicrobial activity showed strong inhibition by old Upper Mountain honey extract (80% methanol extract) with the value of 6.00 ± 0.01 mm at concentration of 100% against gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis). The same trend of phytochemicals content and antimicrobial activity was also observed in absolute methanol extracts. Conclusion: The present results suggested that wild raw honey collected at mangrove and mountain area in Sabah contained a wide range of phytochemical compounds which has the potential for human health

Chemopreventive and chemotherapeutic properties of selected fruits endemic to Borneo: investigation on Mangifera pajang and Artocarpus odoratissimus

Consumption of fruits and vegetables has been shown to reduce the risk of various types of cancer. Macro- and micro-nutrients as well as non-nutritive phytochemicals present in fruits and vegetables have been associated with this effect. This study was conducted to investigate the chemopreventive and chemotherapeutic potential of two types of fruits which are endemic to Borneo Island: Mangifera pajang (bambangan) and Artocarpus odoratissimus (tarap). The first part of the project was to study the antioxidant potential of the crude extracts of the plants in vitro. The fruits were first separated into flesh, kernel and peel for M. pajang and flesh and seed for A. odoratissmus. DPPH (2,2- diphenyl-I-picrylhydrazyl) free radical scavenging and FRAP (ferric reducing / antioxidant power) assay were employed for the antioxidant study. The result showed that the kernel of M. pajang extract displayed strongest antioxidant activity as assessed using both assays, followed by M. pajang peel, A. odoratissmus seed, M. pajang flesh and A. odoratissmus flesh. The presence of selected phytochemicals in the plant extracts was determined in the next chapter. Polyphenols have been identified as major phytochemicals in the plant extracts, and in M. pajang kernel extract represents about 10% of its total weight. Gallic acid, coumaric acid, sinapic acid, caffeic acid, ferulic acid, chlorogenic acid, naringin, hesperidin, rutin, luteolin and diosmin have been identified as the key polyphenol phytochemicals present in the kernel of M. pajang which might be responsible for the superior antioxidant properties as compared to other extracts. Concern that the results for the chemical antioxidant assay do not necessarily reflect cellular activity led to the third part of the project; assessment of the cytoprotective activity of the crude extracts against oxidative stress induced by tert-butyl hydroperoxide (t-BHP). Only M. pajang kernel extract as well as the positive control (quercetin) displayed cytoprotective activity against this toxicant. It seems that non-cell based antioxidant assay does not necessarily reflect the activity in cell-based antioxidant assay. This is shown by lack of cytoprotective activity of both M. pajang peel and A. odoratissimus extracts despite their considerably high antioxidant activity in DPPH free radical scavenging and FRAP assay. In order to study which proteins might be involved in the cytoprotection mechanism, western blotting method was employed to determine the expression of various Cytoprotective proteins [i.e. quinone reductase (NQO I), glutathione peroxidase (OR), methionine sulfoxide reductase A (MSRA), heat shock protein 27 (HSP27) and heat shock protein 70 (HSP 70)]. Different cytoprotective mechanisms were observed by the kernel extracts and quercetin. In the present study, NQOI, OR, MSRA, HSP27 and HSP 70 have been shown to be involved in the cytoprotection activity of quercetin while only OR and MSRA were involved in the cytoprotection activity of M. pajang kernel extracts. Other cytoprotective proteins remain to be studied to fully understand the cytoprotection mechanism of both plant extract and quercetin. Some chemopreventive agents have been shown to suppress cancer proliferation, induce apoptosis in cancer cells as well as inhibit angiogenesis and metastasis in pre-clinical and clinical trials. Thus, the last part of the project was to determine the anti-cancer potential of plant extracts in a variety of cancer cell lines (derived from breast, colon, liver and ovarian carcinoma). The results showed that the kernel extract of M. pajang displayed strong anti proliferative activity in breast cancer cell lines (MCF-7 and MDA-MB-231). The kernel extract induced cell cycle arrest in MCF-7 cells at the sub-G1 (apoptosis) phase of the cell cycle in a time-dependent manner. For MDA-MB-231 cells, the kernel extract induced strong G2-M arrest in cell cycle progression at 24 hours, resulting in substantial sub-G1 (apoptosis) arrest after 48 and 72 hours of incubation. Staining with Annexin V -FITC and propidium iodide revealed that this apoptosis occurred early in both cell types, 36 hours for MCF-7 cells and 24 hours for MDA-MB- 231 cells, with 14.0% and 16.5% of the cells respectively undergoing apoptosis at these times. This apoptosis appeared to be dependent on caspases-2 and -3 in MCF-7 cells and on caspases-2, -3 and -9 in MDA-MB-231 cells. As a conclusion, from the two plants (M. pajang and A. odoratissimus) studied, the extract of M. pajang kernel displayed diverse health benefit properties, antioxidant, chemoprevention and chemotherapeutic potential. M. pajang could be fully utilized for pharmaceutical, nutraceutical as well as food products. Further study (i.e. animal and clinical study, isolation of pure compounds, bioavailability study) are required to determine the efficacy in human population

Traditional vegetable salad (ulam) of Borneo as source of functional food

Traditional vegetable salad or “ulam” are shown to have beneficial properties for health maintenance and should be further studied and used as a source of food as well as medicine. Apparently, the fern species are highlighted common plants that have been consumed as traditional vegetable all around Borneo, especially Stenochlaena palustris or locally known as “lemiding” (Brunei), “lambiding” (Sabah), “midin” (Sarawak), and “kalakai” (Kalimantan). Stenochlaena palustris was also studied for its phenolic contents and antioxidant properties. The extract of the edible young sterile frond of Stenochlaena palustris contained more anthocyanins (51.32 mg/100 g dry matter) compared to extracts of mature sterile, young fertile, and mature fertile fronds of the fern. The Penan people also used Stenochlaena palustris as herbal medicine to reduce high fever and served it to new mothers during the postpartum recovery period. Other fern species that have been cited used in Borneo are Diplazium esculentum, Nephrolepis acutifolia, and Nephrolepis bisserata. Fruit of Solanum torvum, leaves of Cosmos caudatus, the flower buds of Etlingera spp., young shoot of bamboo species, and banana inflorescence have been consumed as traditional vegetable in many parts of Borneo, the preparation being either fresh or used as a condiment in cooking. In previous literature also noted that seaweeds namely Kappahycus alvarezzii and Eucheuma denticulatum, as well as mushrooms like Termitomyces aurantiacus, have also been administered as traditional vegetable. It is scientifically shown that traditional vegetable from Borneo have potential as food that could bring health benefit to the consumer, especially in preventing oxidative damage related diseases, microbial infection, and metabolic disorder such as diabetes. Innovation on consumption method of traditional vegetable also suggested, for example, the traditional vegetable be developed into natural food products and food additive, e.g. herbal teas, essential o

Effects of Strobilanthes crispus tea aqueous extracts on glucose and lipid profile in normal and streptozotocin-induced hyperglycemic rats.

Strobilanthes crispus (Acanthaceae) has been used traditionally as antidiabetic, diuretic, antilytic, and laxative and has been proven scientifically to possess high antioxidant activity, anti-AIDS, and anticancer properties. It is commonly consumed in the form of herbal tea. The ethnopharmacological value of this plant, such as the development of nutraceutical S. crispus herbal tea (fermented and unfermented) and assessment of their antihyperglycemic properties were investigated. The antidiabetic properties of S. crispus fermented and unfermented tea was carried out in normal and streptozotocin-induced hyperglycaemic rats for 21 days. Glucose and lipid profile (total cholesterol, triglyceride, HDL-cholesterol, LDL-cholesterol) were determined at day 0 (baseline), day 7, and day 21. The results showed that the hot water extract of both fermented and unfermented S. crispus tea reduced blood glucose in hyperglycaemic rats. S. crispus unfermented tea also reduced glucose level in normal rat. Both fermented and unfermented S. crispus tea also showed to improve lipid profile. Antioxidant and polyphenol content that present in the extracts might contribute to the antihyperglycemic and antilipidemic properties. Further study is needed to be carried out in pre-clinical and clinical environment to prove its efficacy in human

Influence of Light Intensity on the Photosynthesis and Phenolic Contents of Mangifera Indica

Light is an important environmental factor that have an influence on a plants photosynthesis and production of secondary metabolites like phenolic compounds and flavonoid. Mangifera indica from the family Anacardiaceae is known to have bioactivity due to its phenolic and flavonoid contents. The objective of the study is to determine the influence of light on the photosynthesis and phenolic contents of M. indica. Photosynthesis of the plant was measured using a portable photosynthesis system referred to as LICOR- 6400. Photosynthetic pigments as well as phenolic and flavonoid contents were quantified using a UV-VIS spectrophotometer. The outcome derived from the study shows that sun exposed leaves of the studied plant were having the maximum photosynthesis, saturation and compensation points (P &lt; 0.05).  Moreover, sun exposed leaves were having higher carotenoid, phenolic and flavonoid contents but lower chlorophyll contents. This leads to a conclusion that sun leaves of M. indica contribute the highest photosynthesis and phenolic contents to the plant

Unfermented Freeze-Dried Leaf Extract of Tongkat Ali (Eurycoma longifolia Jack.) Induced Cytotoxicity and Apoptosis in MDA-MB-231 and MCF-7 Breast Cancer Cell Lines

possible anticancer mechanism of action against breast cancer cell lines: non-hormone-dependent MDA-MB-231 and hormonedependent MCF-7. -e leaves of E. longifolia were processed into unfermented and fermented batches before drying using freeze and microwave-oven drying techniques. Obtained extracts were tested for cytotoxicity effect using MTT assay and phenolic determination using HPLC-DAD technique. -e most toxic sample was analyzed for its apoptotic cell quantification, cell cycle distribution, and the expression of caspases and apoptotic protein using flow cytometry technique. Fragmentation of DNA was tested using an agarose gel electrophoresis system. -e results determined that the unfermented freeze-dried leaf extract was the most toxic towards MDA-MB-231 and MCF-7 cells, in a dose-dependent manner. -is extract contains the highest phenolics of gallic acid, chlorogenic acid, ECG, and EGCG. -e DNA fragmentation was observed in both cell lines, where cell cycle was arrested at the G2/M phase in MCF-7 cells and S phase in MDA-MB-231 cells. -e number of apoptotic cells for MDA-MB-231 was increased when the treatment was prolonged from 24 h to 48 h but slightly decreased at 72 h, whereas apoptosis in MCF-7 cells occurred in a time-dependent manner. -ere were significant activities of cytochrome c, caspase-3, Bax, and Bcl-2 apoptotic protein in MDA-MB-231 cells, whereas MCF-7 cells showed significant activities for caspase-8, cytochrome c, Bax, p53, and Bcl-2 apoptotic protein. -ese results indicate the ability of unfermented freeze-dried leaf extract of E. longifolia to induce apoptosis cell death on MDA-MB-231 and MCF-7, as well as real evidence on sample preparation effect towards its cytotoxicity level

Optimization of extraction conditions of phytochemical compounds and anti-gout activity of euphorbia hirta l. (Ara Tanah) using response surface methodology and Liquid Chromatography-Mass Spectrometry (LC-MS) analysis

Gout is a common disease affected most of the people due to the elevation of uric acid in the blood. Flavonoid and phenolic compounds are reported to exert the anti-gout activity of medicinal plants. Hence, this study aimed at optimizing the extraction conditions of phenolic and flavonoid compounds as well as the anti-gout (xanthine oxidase inhibitory activity) in vitro of Euphorbia hirta using response surface methodology (RSM). /e plant part used was the whole plant excluding roots. /e effects of three independent variables (extraction time, X1; extraction temperature, X2; and solid-to-liquid ratio, X3) on three response variables (total flavonoid content, Y1; total phenolic content, Y2; and xanthine oxidase inhibitory activity, Y3) were determined using central composite design (CCD) while phytochemical profiling of the extracts was determined by liquid chromatography-mass spectrometry (LC-MS). Quadratic models produced a satisfactory fitting of the experimental data with regard to total flavonoid content (r2 � 0.9407, p < 0.0001), total phenolic content (r2 � 0.9383, p < 0.0001), and xanthine oxidase inhibitory activity (r2 � 0.9794, p < 0.0001). /e best extraction conditions observed for total flavonoid content, total phenolic content, and xanthine oxidase inhibitory activity were at a temperature of 79.07°C for 17.42 min with solid-to-liquid ratio of 1 : 20 g/ml. /e optimum values for total flavonoid, total phenolic, and xanthine oxidase inhibitory activity were 67.56 mg RE/g, 155.21 mg GAE/g, and 91.42%, respectively. /e main phytochemical compounds in the optimized E. hirta extract are neochlorogenic acid, quercetin-3β-D-glucoside, syringic acid, caffeic acid, ellagic acid, astragalin, afzelin, and quercetin. As conclusion, this study clearly demonstrated the best conditions to obtain higher xanthine oxidase inhibitory activity and phytochemical compounds which can be further used for the development of anti-gout agents

Effect of light intensity on the photosynthesis and stomatal density of selected plant species of Gunung Ledang, Johor

Light intensity influences the photosynthesis and stomatal density in plants. In this study, it was hypothesized that if the transpiration rate of a plant is low, then the stomatal density might be high. The objective of this research is to investigate the effect of varying light intensity on the photosynthetic capacity and stomatal density of Mikania micrantha, Tridax procumbens, Clidemia hirta and Centella asiatica plant species of Gunung Ledang, Johor, Malaysia. Sun-exposed and shaded species were selected for the study. Gas exchange characteristics including stomatal conductance to water vapour (gsw), transpiration rate and intracellular CO2 (Ci) were measured using LI-6400 portable photosynthesis system while net assimilation rate (Anet), light saturation point (LSP), light compensation point (LCP), and apparent quantum yield (Aqy) were calculated from the photosynthetic light response curve. Stomatal density was determined using nail polish impression method and examined using a stereo microscope. The result of this study shows that high light intensity influences the net assimilation rate and stomatal density of the studied plants. As the light intensity increases, Amax, LSP, LCP and stomatal density of the plants increases. There is a significant difference between the light response characteristics and stomatal density of the sun-exposed and shaded species (P < 0.05). A statistically significant negative correlation (P < 0.05) was achieved among stomatal density and transpiration rate. As stomatal density increases, the transpiration rate decreases. The result leads to a conclusion that Mikania micrantha can efficiently utilise the amount of light available more than Tridax procumbens, Clidemia hirta and Centella asiatica and grow abundantly as dominant species in an introduced environment

Cytotoxic properties of selected Etlingera spp. and Zingiber spp. (Zingiberaceae) endemic to Borneo

Zingiberaceae are known as valuable herbs with an important role in the prevention and treatment of various diseases. More than 300 species of Zingiberaceae were documented in Borneo. In this study, methanolic extracts of three species of Zingiberaceae (namely, Etlingera velutina, Etlingera belalongensis and Zingiber vinosum) were analysed for their total phenolic and flavonoid contents and cytotoxic activity in vitro. The cytotoxic activities of these extracts were tested against several cancer cell lines, such as hormone dependent breast cancer (MCF-7), non-hormone dependent cancer (MDA-MB-231), ovarian cancer (CaOV3) and cervical cancer (Hela) using MTT assay. Crude extracts from rhizome of E. belalongensis and E. velutina showed significant cytotoxic activity against MDA-MB-231 cell line proliferation, with IC50 values (concentration which inhibit 50% of cell population) of 51.00±4.24 µg/ml and 67.00±9.89 µg/ml, respectively. The methanol extracts were further analysed for the cell cycle analysis using flow cytometry. The results showed that the Etlingera species exhibited higher antioxidant activity and stronger cytotoxic activity in selected cancer cell lines, with the highest cell death accumulated in G1 phase as compared to Zingiber species. Thus, polyphenol phytochemicals could be the major contributors to the cytotoxic activity of these species. As a conclusion, tropical gingers in Borneo investigated in this study have the potential to be developed as anticancer remedies