Botanical Origin Differentiation of Malaysian Stingless Bee Honey Produced by Heterotrigona itama and Geniotrigona thoracica Using Chemometrics.

Stingless bee honey, specifically honeydew honey, is generally valued for its better health benefits than those of most blossom types. However, scientific studies about the differentiation of stingless bee honey based on honeydew and blossom origins are very limited. In this study, 13C NMR spectroscopy was employed to quantify the seven major sugar tautomers in stingless bee honey samples, and the major sugar compositions of both honeydew and blossom types were found not significantly different. However, several physicochemical properties of honeydew honey including moisture content, free acidity, electrical conductivity, ash content, acetic acid, diastase, hydrogen peroxide, and mineral elements levels were significantly higher; while total soluble solid, proline, and hydroxymethylfurfural were significantly lower than blossom honey. Greater antioxidant capacity in honeydew honey was proven with higher total phenolic compounds, ABTS, DPPH, superoxide radical scavenging activities, peroxyl radical inhibition, iron chelation, and ferric reducing power. Using principal component analysis (PCA), two clusters of stingless bee honey from the honeydew and blossom origin were observed. PCA also revealed that the differentiation between honeydew and blossom origin of stingless bee honey is possible with certain physicochemical and antioxidant parameters. The combination of NMR spectroscopy and chemometrics are suggested to be useful to determine the authenticity and botanical origin of stingless bee honey.

Antifungal and cytotoxic activities of selected medicinal plants from Malaysia.

This study was conducted to investigate the antifungal potential and cytotoxicity of selected medicinal plants from Malaysia. The extracts from the stem of Cissus quadrangularis and the leaves of Asplenium nidus, Pereskia bleo, Persicaria odorata and Sauropus androgynus were assayed against six fungi using p-iodonitrotetrazolium-based on colorimetric broth microdilution method. All the plant extracts were found to be fungicidal against at least one type of fungus. The strongest fungicidal activity (minimum fungicidal concentration=0.16 mg/mL) were exhibited by the hexane extract of C. quadrangularis, the hexane, chloroform, ethanol and methanol extracts of P. bleo, the hexane and ethyl acetate extracts of P. odorata, and the water extract of A. nidus. In terms of cytotoxicity on the African monkey kidney epithelial (Vero) cells, the chloroform extract of P. odorata produced the lowest 50% cytotoxic concentration (100.3 ± 4.2 µ g/mL). In contrast, none of the water extracts from the studied plants caused significant toxicity on the cells. The water extract of A. nidus warrants further investigation since it showed the strongest fungicidal activity and the highest total activity (179.22 L/g) against Issatchenkia orientalis, and did not cause any toxicity to the Vero cells.

Bioactivity-guided isolation and structural characterization of the antifungal compound, plumbagin, from Nepenthes gracilis.

CONTEXT: Despite several phytochemical studies of Nepenthes gracilis Korth (Nepenthaceae), the biological activities of this pitcher plant remain to be explored. OBJECTIVE: This study evaluates the antifungal activity of N. gracilis extracts, isolates, and characterizes its bioactive compound and evaluates the cytotoxicity of the isolated compound. MATERIALS AND METHODS: Fresh leaves of N. gracilis were sequentially extracted. The fungistatic and fungicidal activities of the extracts were evaluated against six species of fungi of medical importance using a colorimetric broth microdilution method. The most active extract was fractionated by liquid-liquid partitioning and further purified by a preparative thin layer chromatography. Structural elucidation was carried out using FT-IR, GC-MS, and NMR. Cytotoxicity testing against rhesus monkey kidney epithelial cells (LLC-MK2) was assessed by a neutral red uptake (NRU) assay. RESULTS: The hexane extract, which showed the lowest minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC), both at 20 µg/mL against Candida albicans, Issatchenkia orientalis, and Trichophyton mentagrophytes, was subjected to bioactivity-guided fractionation. The isolated compound exhibited potent activity with the MIC values ranging from 2 to 31 µg/mL against all the fungi. The active compound was identified as plumbagin (5-hydroxy-2-methyl-naphthalene-1,4-dione). The 50% cytotoxicity concentration (CC50) of plumbagin was 0.60 µg/mL. DISCUSSION AND CONCLUSION: The selectivity indices of plumbagin against all the fungi were less than 1.0, indicating that plumbagin is more toxic to mammalian than fungal cells. This study provides information on the antifungal properties of N. gracilis leaf extracts, as well as the antifungal and cytotoxicity properties of plumbagin.

Mosquito Larvicidal Activity and Cytotoxicity of the Extracts of Aromatic Plants from Malaysia.

Despite ongoing control efforts, the mosquito population and diseases vectored by them continue to thrive worldwide, causing major health concerns. There has been growing interest in the use of botanicals as alternatives to insecticides due to their widespread insecticidal properties, biodegradability, and adaptability to ecological conditions. In this study, we investigated the larvicidal activity and cytotoxicity effects of solvent extracts from three aromatic plants-Curcuma longa (turmeric), Ocimum americanum (hoary basil), and Petroselinum crispum (parsley)-against Aedes albopictus. Subsequently, we examined the phytochemical composition of the extracts through gas chromatography-mass spectrometry (GC-MS) analysis. Results revealed that the hexane extracts of O. americanum and P. crispum exhibited the greatest larvicidal activity with the lowest median lethal concentration (LC50) values (<30 µg/mL) at 24 h post-treatment, with the former found to be significantly less toxic towards African monkey kidney (Vero) cells. The GC-MS analysis of the said extract indicated the presence of different classes of metabolites, including phenylpropanoids, very long-chain alkanes, fatty acids and their derivatives, and terpenes, with the most abundant component being methyl eugenol (55.28%), most of which, have been documented for their larvicidal activities. These findings provide valuable insights into the potential use and development of bioinsecticides, particularly from O. americanum.

Effect of Alpha-S1-Casein Tryptic Hydrolysate and L-Theanine on Poor Sleep Quality: A Double Blind, Randomized Placebo-Controlled Crossover Trial.

This randomized, placebo-controlled, crossover and double-blind study investigates the effects of RLX2™ containing alpha-s1-casein tryptic hydrolysate and L-theanine in working adults affected by poor sleep quality. The supplement or placebo was randomly and blindly assigned to 39 subjects for four weeks and the changes in the subjective sleep assessment via the Pittsburgh Sleep Quality Index (PSQI), heart rate, blood pressure, salivary cortisol by high-performance liquid chromatography method and alpha power of awake electroencephalogram (EEG) were studied. The data were analyzed in two ways, by crossover and crossover summed up. The latter depicted that RLX2™ improved PSQI total score, sleep latency, sleep duration, sleep habitual efficiency, daytime dysfunction, and increased total and frontal alpha power significantly (p < 0.05). The supplement prolonged the total sleeping time by 45 min in the supplement receiving group compared to the placebo group (p < 0.001). However, only sleep duration and sleep habitual efficiency showed a profound effect in both analyses (p < 0.05). In conclusion, being given its beneficial effects without notable adverse events, it would be advantageous to use these nutraceutical ingredients to promote better sleep quality. Further studies with a larger number participants are warranted to support these findings.

Chemical composition, antioxidant, antimicrobial and antiviral activities of the leaf extracts of Syzygium myrtifolium.

This study was conducted to evaluate the chemical composition and biological activities of the leaf extracts of Syzygium myrtifolium Walp. (Myrtaceae). The results indicate that the leaf extracts of S. myrtifolium contain various classes of phytochemicals (alkaloids, anthraquinones, flavonoids, phenolics, saponins, tannins and triterpenoids) and possess antioxidant, antibacterial, antifungal and antiviral activities. Ethyl acetate, ethanol, methanol, and water extracts exhibited significantly higher (p < 0.05) oxygen radical absorbance capacity and ferric-reducing antioxidant power than the hexane and chloroform extracts. However, all extracts exhibited stronger inhibitory activity against four tested species of yeasts (minimal inhibitory concentration: 0.02-0.31 mg mL-1) than against six tested species of bacteria (minimal inhibitory concentration: 0.16-1.25 mg mL-1). The ethanolic extract offered the highest protection of Vero cells (viability > 70 %) from the cytopathic effect caused by the Chikungunya virus while the ethyl acetate extract showed significant replication inhibitory activity against the virus (p < 0.001) using the replicon-enhanced green fluorescent protein reporter system.

Mode-Dependent Antiviral Activity of Medicinal Plant Extracts against the Mosquito-Borne Chikungunya Virus.

The lack of specific treatment for chikungunya fever makes the need for anti-chikungunya virus agents more crucial. This study was conducted to evaluate 132 extracts obtained by sequential solvent extraction from 21 medicinal plants for cytopathic effect inhibitory activity using virus-infected Vero cells in two different sample introduction modes. Among the extracts, 42 extracts (31.8%) from 12 plants in the concurrent mode and three extracts (2.3%) from a plant in the non-concurrent mode displayed strong cytopathic effect inhibitory activity (cell viability ≥70%). Viral load quantification analysis unveiled that the extracts of Clinacanthus nutans (chloroform, ethyl acetate, and ethanol), Hydrocotyle sibthorpioides (ethanol), and Ocimum americanum (ethanol and methanol) hindered the release of viral progeny from the infected cells while the extracts of Ficus deltoidea (ethanol), Gynura bicolor (water), H. sibthorpioides (water), and O. americanum (chloroform and ethyl acetate) blocked the entry of virus into the cells. The extracts of Diodella sarmentosa (ethyl acetate), Diplazium esculentum (chloroform, ethyl acetate, and ethanol), and G. bicolor (ethanol) possessed virucidal effect and caused 5.41-log to 6.63-log reductions of viral load compared to the virus control. The results indicate that these medicinal plants are potential sources of anti-chikungunya virus agents that have varied modes of action.

Self-management using crude herbs and the health-related quality of life among adult patients with hypertension living in a suburban setting of Malaysia.

PURPOSE: To determine the prevalence of crude herbs' use in the self-management of hypertension and the health-related quality of life (HRQOL) in patients with hypertension. METHODS: This cross-sectional study was performed among patients with hypertension attending a government health clinic. Socio-demographic characteristics, lifestyle modifications, medical history and predictors of crude herbs users were obtained. The diversity of crude herbs used was assessed using a modified international complementary and alternative medicine questionnaire (I-CAM-Q) and the HRQOL was assessed using the SF36 instrument. RESULTS: Out of the 294 patients recruited, 52.4% were female, 41.5% were Malay and 38.8% were within the 60 to69 age category. The prevalence of crude herbs users was 30.6% and the most common herbs used were pegaga (Centella asiatica), peria (Momordica charantia) and betik (Carica papaya). Using the regression analysis, significantly higher odds of using crude herbs are noted among Malay or Indian patients who have these characteristics: attained secondary education, experienced falls or muscle pain, and had systolic blood pressure of more than 140 mmHg. There was no significant difference in HRQOL domains between the crude herb users and non-users (p>0.05). CONCLUSION: Besides taking allopathic medications, certain patients with hypertension use crude herbs as a form of self-management. Although patients are adamant about integrating crude herbs as a form of self-management, the effects of doing so have not been properly investigated. This implies that the healthcare staff members need to communicate with the patients regarding the use of crude herbs together with conventional drugs.

The Antibacterial Potential of Honeydew Honey Produced by Stingless Bee (Heterotrigona itama) against Antibiotic Resistant Bacteria.

Scientific studies about the antibacterial effects of honeydew honey produced by the stingless bee are very limited. In this study, the antibacterial activities of 46 blossom and honeydew honeys produced by both honey bees and stingless bees were evaluated and compared. All bacterial isolates showed varying degrees of susceptibility to blossom and honeydew honeys produced by the honey bee (Apis cerana) and stingless bee (Heterotrigona itama and Geniotrigona thoracica) in agar-well diffusion. All stingless bee honeys managed to inhibit all the isolates but only four out of 23 honey bee honeys achieved that. In comparison with Staphylococcus aureus, Escherichia coli was found to be more susceptible to the antibacterial effects of honey. Bactericidal effects of stingless bee honeys on E. coli were determined with the measurement of endotoxins released due to cell lysis. Based on the outcomes, the greatest antibacterial effects were observed in honeydew honey produced by H. itama. Scanning electron microscopic images revealed the morphological alteration and destruction of E. coli due to the action of this honey. The combination of this honey with antibiotics showed synergistic inhibitory effects on E. coli clinical isolates. This study revealed that honeydew honey produced by H. itama stingless bee has promising antibacterial activity against pathogenic bacteria, including antibiotic resistant strains.

Investigation of twenty selected medicinal plants from Malaysia for anti-Chikungunya virus activity.

Chikungunya virus is a reemerging arbovirus transmitted mainly by Aedes mosquitoes. As there are no specific treatments available, Chikungunya virus infection is a significant public health problem. This study investigated 120 extracts from selected medicinal plants for anti-Chikungunya virus activity. The plant materials were subjected to sequential solvent extraction to obtain six different extracts for each plant. The cytotoxicity and antiviral activity of each extract were examined using African monkey kidney epithelial (Vero) cells. The ethanol, methanol and chloroform extracts of Tradescantia spathacea (Commelinaceae) leaves showed the strongest cytopathic effect inhibition on Vero cells, resulting in cell viabilities of 92.6% ± 1.0% (512 µg/ml), 91.5% ± 1.7% (512 µg/ml) and 88.8% ± 2.4% (80 µg/ml) respectively. However, quantitative RT-PCR analysis revealed that the chloroform extract of Rhapis excelsa (Arecaceae) leaves resulted in the highest percentage of reduction of viral load (98.1%), followed by the ethyl acetate extract of Vernonia amygdalina (Compositae) leaves (95.5%). The corresponding 50% effective concentrations (EC50) and selectivity indices for these two extracts were 29.9 ± 0.9 and 32.4 ± 1.3 µg/ml, and 5.4 and 5.1 respectively. Rhapis excelsa and Vernonia amygdalina could be sources of anti-Chikungunya virus agents. [Int Microbiol 19(3):175-182 (2016)].

High performance liquid chromatography profiling of health-promoting phytochemicals and evaluation of antioxidant, anti-lipoxygenase, iron chelating and anti-glucosidase activities of wetland macrophytes.

BACKGROUND: The phytochemistry and bioactivity of wetland macrophytes are underexplored. Plants are known as the natural sources of phytochemical beneficial to health. OBJECTIVE: The objective of this study is to analyze the phytochemical profiles and bioactivities of 10 extracts prepared from different plant parts of wetland macrophytes Hanguana malayana, Ludwigia adscendens and Monochoria hastata. MATERIALS AND METHODS: High performance liquid chromatography (HPLC) was used to analyze the phytochemical profile of the extracts. Antioxidant assay such as 2,2-diphenyl-1-picrylhydrazyl, nitric oxide (NO) radical scavenging activity and ferric reducing antioxidant power were performed. Bioactivity assays carried out were anti-lipoxygenase, anti-glucosidase, and iron chelating. RESULTS: Leaf extract of L. adscendens had the highest 2,2-diphenyl-1-picrylhydrazyl (half of maximal effective concentration [EC50] =0.97 mg/mL) and NO (EC50 = 0.31 mg/mL) scavenging activities. The extract also exhibited the highest iron chelating (EC50 = 3.24 mg/mL) and anti-glucosidase (EC50 = 27.5 µg/mL) activities. The anti-glucosidase activity of L. adscendens leaf extract was comparable or superior to those of acarbose, myricetin and quercetin. Correlation between iron chelating and radical scavenging activities among the extracts implies the presence of dual-function phytoconstituents with concurrent iron chelating and radical scavenging activities. HPLC analysis revealed the presence of p-coumaric acid (p-CA), gallic acid (GA) and myricetin in all or most extracts. M. hastata fruit and leaf extracts had the highest p-hydroxybenzoic acid content. Antioxidant and anti-glucosidase activities of the extracts were correlated with p-CA, GA, and myricetin contents. CONCLUSION: Our study demonstrated that wetland macrophytes H. malayana, L. adscendens and M. hastata are potential sources of health-promoting phytochemicals with potent therapeutically-relevant bioactivities.