The aim of this study was to evaluate the genotoxic effects of a crude extract of khat (Catha edulis, Forsk) leaves in rats. Two groups were fed khat crude extract, 1000 and 2000mg/kg body weight, for 90 days and were compared with a control group. The alkaline (pH>13) version of comet assay was used in this study. However, no previous published work has been undertaken and showed the effect of khat on DNA migration in the comet assay. To compare the comet assay results with another genetic endpoint, blood samples were analyzed for chromosomal aberrations. These results showed no DNA damage detected using comet assay in both the khat treated groups, while the results of chromosomal aberrations assay showed a significant increase (P<0.05) in the 2000mg/kg body weight treated group compared to the control group.
Methanol extract of the Gracilaria changii has been screened for antimicrobial activity against Pseudomonas aeruginosa. Antimicrobial activities were carried out using disc diffusion assay and broth dilution method against P. aeruginosa. The methanol extract of G. changii showed a good antimicrobial activity against P. aeruginosa with MIC (Minimum Inhibitory Concentration) value of 6.25mg/ml. Exposure of P. aeruginosa cells to 6.25mg/ml of methanol extract of G. changii resulted in complete inhibition of the bacterial cells. The main abnormalities noted via SEM and TEM studies were the alterations in morphology and cytology of the bacterial cells. The main reason for this deterioration was discussed. The effect of the methanol extract on the growth profile for the bacteria was also done and confirmed the bactericidal effect of the G. changii methanol extract on P. aeruginosa by changing the normal growth profile of P. aeruginosa. In an acute toxicity study using mice, the median lethal dose (LD(50)) of the extract was greater than 2000 mg/kg, and we found no pathological changes in macroscopic examination by necropsy of mice treated with extract. We conclude that G. changii might be safely used as an antimicrobial agent.
This study was conducted to evaluate the effects of topical application of aqueous extract of Hericium erinaceus fruiting bodies (HEFB) on the rate of wound healing enclosure and histology of the healed wound. Five groups of male Sprague-Dawley rats were experimentally wounded in the posterior neck area. A uniform wound area of 2.00 cm in diameter, using a circular stamp, was excised from the nape of the dorsal neck of all rats with the aid of a round seal. The animal groups were topically treated, respectively, with 0.2 mL each of sterilized distilled water (sdH2O); Intrasite gel; and 20, 30, and 40 mg/mL HEFB. Macroscopically, those rats whose wounds were dressed with HEFB and those in the Intrasite gel-treated group healed earlier than those treated with sdH2O. Histological analysis of healed wounds dressed with HEFB showed less scar width at wound enclosure and the healed wound contained fewer macrophages and more collagen with angiogenesis, compared to wounds dressed with sdH2O. In conclusion, wounds dressed with HEFB significantly enhanced the acceleration of wound healing enclosure in rats.
The present study aims to assess the antioxidant activities (AOA) and total phenolic content (TPC) of water extracts of selected edible wild mushrooms: Pleurotus porrigens, Schizophyllum commune, Hygrocybe conica, and Lentinus ciliatus. The AOA were evaluated against DPPH radical and ABTS radical cation scavenging ability, ferric-reducing antioxidant power (FRAP) and beta-carotene-linoleate bleaching (beta-CB) assays, and the Folin-Ciocalteu method for TPC. BHA was used as reference. P. porrigens showed significantly higher (p < 0.05) DPPH* scavenging ability (90.78 +/- 0.30%) and FRAP (6.37 +/- 0.22 mM FE/100g), while Sch. commune showed significantly higher (p < 0.05) ABTS*+ inhibition activity (94.96 +/- 0.70%) and beta-CB inhibition activity (94.18 +/- 0.17%), respectively. TPC was found in a descending order of P. poriggens > L. ciliatus = Pleurotus ostreatus (cultivated) > H. conica = Sch. commune. Positive correlation was observed between the AOA and TPC. When compared to BHA (2 mM), P. porrigens showed significantly higher (p < 0.05) DPPH* scavenging ability and reducing power, while Sch. commune showed comparable DPPH* scavenging ability and ABTS*+ inhibition activity. All the mushrooms have better ABTS*+ inhibition activity than BHA (1 mM). The beta-CB inhibition activity of BHA was significantly higher than those of edible wild mushrooms. The water extracts of edible wild mushrooms showed potent antioxidant activities compared to BHA to a certain extent.
The antiplasmodial activity of the crude extracts of thirty plant species collected from Sabah was evaluated using chloroquine-sensitive strain (D10) and chloroquine-resistant strain (Gombak A) of Plasmodium falciparum. Significant activities were observed for the bark extract of Polyalthia insignis (IC50 3.89 and 11.89 mg/ml against Gombak A and D10, respectively), the leaf extracts of Kopsia dasyrachis (4.62 mg/ml against Gombak A) and Litsea elliptibacea (IC50 8.88 mg/ml against Gombak A), as well as the leaf and bark extracts of Neouvaria acuminatissima (IC50 6.90-10.08 and 0.69 mg/ml against Gombak A and D10, respectively), and the bark extract of Polyalthia microtus(IC50 9.0 and 12.12 mg/ml against Gombak A and D10, respectively).
A binary mixture of starch-starch or starch with other biopolymers such as protein and non-starch polysaccharides could provide a new approach in producing starch-based food products. In the context of food processing, a specific adjustment in the rheological properties plays an important role in regulating production processing and optimizing the applicability, stability, and sensory of the final food products. This review examines various biopolymer mixtures based on starch and the influence of their interaction on physicochemical and rheological properties of the starch-based foods. It is evident that the physicochemical and rheological characteristics of the biopolymers mixture are highly dependent on the type of starch and other biopolymers that make them up mixing ratios, mixing procedure and presence of other food ingredients in the mixture. Understanding these properties will lead to improve the formulation of starch-based foods and minimize the need to resort to chemically modified starch.
We present a model case study of the activity of aqueous extract of Hericium erinaceus fresh fruit bodies in promoting functional recovery following crush injury to the peroneal nerve in adult female Sprague-Dawley rats. The aim was to explore the possible use of this mushroom in nerve repair. The activities of aqueous extract were compared to activities exhibited by mecobalamin (vitamin B12), which has been widely used in the treatment of peripheral nerve disorders. Analysis of walking track indicated that return of hind limb function and normal toe spreading occurred earlier in treated groups than in the negative control (non-treated) group. Regeneration of axons and reinnervation of motor endplates/neuromuscular junction in extensor digitorum longus muscle of rats in treated groups developed better than in the negative control group. Further, immunofluorescence studies also showed that dorsal root ganglia neurons ipsilateral to the crush injury in rats of treated groups expressed higher immunoreactivities for Akt and MAPK signaling pathways as well as c-Jun and c-Fos genes compared to the negative control group. Akt cascade plays a major role in mediating neurotrophin-promoted cell survival, while MAPK cascade is involved in mediating neurite outgrowth. Immediate early gene expression was also involved in the cascade of events leading to regeneration. Local axonal protein synthetic machinery was also enhanced in the distal segments of crushed nerves in treated groups. Therefore, daily oral administration of H. erinaceus could promote the regeneration of injured rat peroneal nerve in the early stage of recovery.
Marine fungus Fusarium proliferatum derived from marine sponge collected along Pulau Tinggi, Malaysia was cultivated on Potato Dextrose Broth and incubated for 7 days at 30oC. The liquid cultures were then extracted using ethyl acetate. The crude extract was investigated for its anti-microbial activity and was passed through Sephadex column and the fractions were collected. Reverse phase HPLC was used to monitor the component of crude extract. HPLC guided purification of crude extract resulted in the isolation of linoleic acid, 4-hydroxy phenethyl alcohol, 2,5-furandimethanol and adenosine. Their structures were elucidated by spectroscopic methods.
An endophytic fungus isolated from the plant Cinnamomum mollissimum was investigated for the bioactivity of its metabolites. The fungus, similar to a Phoma sp., was cultured in potato dextrose broth for two weeks, followed by extraction with ethyl acetate. The crude extract obtained was fractionated by high-performance liquid chromatography. Both crude extract and fractions were assayed for cytotoxicity against P388 murine leukemic cells and inhibition of bacterial and fungal pathogens. The bioactive extract fraction was purified further and characterized by nuclear magnetic resonance, mass spectral and X-ray crystallography analysis. A polyketide compound, 5-hydroxyramulosin, was identified as the constituent of the bioactive fungal extract fraction. This compound inhibited the fungal pathogen Aspergillus niger (IC(50) 1.56 μg/mL) and was cytotoxic against murine leukemia cells (IC(50) 2.10 μg/mL). 5-Hydroxyramulosin was the major compound produced by the endophytic fungus. This research suggests that fungal endophytes are a good source of bioactive metabolites which have potential applications in medicine.
Hepato- and nephrotoxicity of Khat consumption (Catha edulis Forskal) have been evoked. Therefore, this study was conducted to evaluate such possible hepatorenal toxicity in female and male Sprague-Dawley rats (SD rats) focusing primarily on liver and kidney. In addition, female and male rats were investigated separately. Accordingly, forty-eight SD-rats (100-120 g) were distributed randomly into four groups of males and female (n = 12). Normal controls (NCs) received distilled water, whereas test groups received 500 mg/kg (low dose (LD)), 1000 mg/kg (medium dose (MD)), or 2000 mg/kg (high dose (HD)) of crude extract of Catha edulis orally for 4 weeks. Then, physical, biochemical, hematological, and histological parameters were analyzed. Results in Khat-fed rats showed hepatic enlargement, abnormal findings in serum aspartate aminotransferase (AST), and alkaline phosphatase (ALP) of male and female SD-rats and serum albumin (A) and serum creatinine (Cr) of female as compared to controls. In addition, histopathological abnormalities confirmed hepatic and renal toxicities of Khat that were related to heavy Khat consumption. In summary, Khat could be associated with hepatic hypertrophy and hepatotoxicity in male and female SD-rats and nephrotoxicity only in female SD-rats.
The sclerotium of the "tiger's milk mushroom" (Lignosus rhinocerotis) is used as tonic and folk medicine for the treatment of cancer, fever, cough and asthma by the local and indigenous communities. It is traditionally prepared by either boiling or maceration-like methods; however, there is no attempt to understand how different processing methods might affect their efficacies as anticancer agents.
Fourier transform infrared spectroscopy (FTIR) combined with multivariate calibration of partial least square (PLS) was developed and optimized for the analysis of Nigella seed oil (NSO) in binary and ternary mixtures with corn oil (CO) and soybean oil (SO). Based on PLS modeling performed, quantitative analysis of NSO in binary mixtures with CO carried out using the second derivative FTIR spectra at combined frequencies of 2977-3028, 1666-1739, and 740-1446 cm(-1) revealed the highest value of coefficient of determination (R (2), 0.9984) and the lowest value of root mean square error of calibration (RMSEC, 1.34% v/v). NSO in binary mixtures with SO is successfully determined at the combined frequencies of 2985-3024 and 752-1755 cm(-1) using the first derivative FTIR spectra with R (2) and RMSEC values of 0.9970 and 0.47% v/v, respectively. Meanwhile, the second derivative FTIR spectra at the combined frequencies of 2977-3028 cm(-1), 1666-1739 cm(-1), and 740-1446 cm(-1) were selected for quantitative analysis of NSO in ternary mixture with CO and SO with R (2) and RMSEC values of 0.9993 and 0.86% v/v, respectively. The results showed that FTIR spectrophotometry is an accurate technique for the quantitative analysis of NSO in binary and ternary mixtures with CO and SO.
This paper presents novel research on the development of a generic intelligent oil fraction sensor based on Electrical Capacitance Tomography (ECT) data. An artificial Neural Network (ANN) has been employed as the intelligent system to sense and estimate oil fractions from the cross-sections of two-component flows comprising oil and gas in a pipeline. Previous works only focused on estimating the oil fraction in the pipeline based on fixed ECT sensor parameters. With fixed ECT design sensors, an oil fraction neural sensor can be trained to deal with ECT data based on the particular sensor parameters, hence the neural sensor is not generic. This work focuses on development of a generic neural oil fraction sensor based on training a Multi-Layer Perceptron (MLP) ANN with various ECT sensor parameters. On average, the proposed oil fraction neural sensor has shown to be able to give a mean absolute error of 3.05% for various ECT sensor sizes.
BACKGROUND: In our previous study, the aqueous extract of Channa striatus (family: Channidae) fillet (AECSF) showed an antidepressant-like effect in mice. However, the mechanism of the antidepressant-like effect is unknown.
AIM: The objective of this study was to explore the involvement of monoamines in the antidepressant-like effect of AECSF in mice.
MATERIALS AND METHODS: AECSF was prepared by steaming the fillets of C. striatus. The male ICR mice were pretreated with various monoaminergic antagonists viz., p-chlorophenylalanine (100 mg/kg, i.p.), prazosin (1 mg/kg, i.p.) and yohimbine (1 mg/kg, i.p.), SCH23390 (0.05 mg/kg, s.c.) and sulpiride (50 mg/kg, i.p.) followed by treatment with AECSF and tested in tail suspension test (TST). Two-way ANOVA with Tukey test were used at p < 0.05 for significance.
RESULTS: The pretreatments with p-chlorophenylalanine, prazosin and yohimbine, but not with SCH23390 and sulpiride, were able to reverse the antidepressant-like effect of AECSF in TST.
CONCLUSIONS: The antidepressant-like effect of AECSF may be mediated through the serotonergic and noradrenergic systems and not through the dopaminergic system.
Proteases in ginger rhizome have the potentials in industrial applications. This study was conducted to extract and characterize the proteolytic enzyme from ginger (Zingiber officinale Roscoe). Ginger protease (GP) was extracted from ginger rhizome by homogenization with 100 mM potassium phosphate buffer pH 7.0 containing 10 mM cysteine and 5 mM EDTA which were found to be the most efficient extraction buffer and stabilizers. After centrifugation at 10,500 x g, protein in the crude extract was precipitated using 60% ammonium sulfate following which the precipitate was redissolved in 50 mM potassium phosphate buffer pH 7.0, dialyzed and then lyophilized. The extraction method yielded 0.94% (w/w of fresh weight) of GP with a specific activity of 27.6 ± 0.1 Unit/mg protein where 1 Unit is defined as the amount of protease causing an increase in absorbance by 1 unit per minute using azocasein as the substrate. Results show that the GP was completely inhibited by heavy metal cations i.e. Cu2+and Hg2+, and a thiol blocking agent or inhibitor, n-ethyl maleimide (NEM), indicating that GP is most probably a cysteine protease. The enzyme has an optimum temperature at 60⁰C and the optimum pH ranged between pH 6 to 8. Monovalent cations (K+ and Na+) have no significant effect on activity of GP, but divalent and trivalent cations showed moderate inhibitory effect. Detergents such as sodium dodecyl sulfate increased the activity of GP while Tween 80 and Tween 20 slightly reduced the activity.
The effects of ethanol concentration (0-100%, v/v), extraction time (60-300 min) and extraction temperature (25-65°C) on the extraction of phenolic antioxidants from Andrographis paniculata was evaluated using single-factor experiments. The following complementary assays were used to screen the antioxidant properties of the crude extracts: total phenolic content (TPC), total flavonoid content (TFC), condensed tannin content (CTC), 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical-scavenging capacity and 2,2’-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging capacity. The extraction conditions chosen had significant effects (p < 0.05) on the extraction of phenolic compounds and antioxidant capacity. The optimal conditions were 60% ethanol for 60 min at 65oC for phenolic compounds and at 25oC for antioxidant capacity. Strong negative significant (p < 0.05) correlations were observed between the phenolic compounds (TPC, TFC and CTC) and antioxidant capacity comprising ABTS (-0.924, -0.909, -0.887, respectively) and DPPH radical-scavenging capacities (-0.992, -0.938, -0.928, respectively) were determined under the influence of extraction temperature.
Tinospora crispa and Tabernaemontana corymbosa have been used traditionally to treat fever, diabetes, rheumatism and sinusitis. The objective of this study was to evaluate the antioxidant activity of Tinospora crispa and Tabernaemontana corymbosa. The presence of apigenin and magnoflorine was detected using LCMS/MS in Tinospora crispa (Patawali) whereas appararicine, voafinine, conodusarine, conodurine, voacamine and voacangine were detected in Tabernaemontana corymbosa (Susur kelapa) methanol extract. The stem extract of Tinospora crispa showed high antioxidant activity in the following order: DPPH radical scavenging, reducing power and metal chelating assay (98.8%, 0.957, 81.97%) than Tabernaemontana corymbosa of leaves (90.04%, 0.652, 69.64%), stem (82.78%, 0.819, 36.70%) and root extracts (63.25%, 0.469, 51.56%), respectively. The high antioxidant activity in the stem extract of Tinospora crispa is due to the presence of apigenin and magnoflorine. The high antioxidant activity in Tabernaemontana corymbosa extract is due to its high phenol contents. There were significant linear positive correlation (r=0.788, p<0.001, r2=0.621) between the total phenolic content and DPPH free radical scavenging assay in the crude extracts of Tinospora crispa and Tabernaemontana corymbosa. Meanwhile, a significant moderate positive correlation was observed between the total phenolic content and ferric reducing power assay (r= 0.556, p<0.05, r2= 0.309). However, there was no significant difference in the correlation coefficient of total phenolic content and metal chelating assay.
The present paper focused on antioxidant and cytotoxicity assessment of crude and total saponin fraction of Chlorophytum borivilianum as an important medicinal plant. In this study, three different antioxidant activities (2,2-diphenyl-1-picrylhydrazyl radical scavenging (DPPH), ferrous ion chelating (FIC), and β -carotene bleaching (BCB) activity) of crude extract and total saponin fraction of C. borivilianum tubers were performed. Crude extract was found to possess higher free radical scavenging activity (ascorbic acid equivalents 2578 ± 111 mg AA/100 g) and bleaching activity (IC50 = 0.7 mg mL(-1)), while total saponin fraction displayed higher ferrous ion chelating (EC50 = 1 mg mL(-1)). Cytotoxicity evaluation of crude extract and total saponin fraction against MCF-7, PC3, and HCT-116 cancer cell lines using 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) cell viability assay indicated a higher cytotoxicity activity of the crude extract than the total saponin fraction on all cell lines, being most effective and selective on MCF-7 human breast cancer cell line.
Amauroderma rugosum is a wild mushroom that is worn as a necklace by the indigenous communities in Malaysia to prevent fits and incessant crying by babies. The aim of this study was to investigate the nutritive composition and antioxidant potential and anti-inflammatory effects of A. rugosum extracts on LPS-stimulated RAW264.7 cells. Nutritional analysis of freeze-dried mycelia of A. rugosum (KUM 61131) from submerged culture indicated a predominant presence of carbohydrates, proteins, dietary fibre, phosphorus, potassium, and sodium. The ethanol crude extract (EE), its hexane (HF), ethyl acetate (EAF), and aqueous (AF) fractions of mycelia of A. rugosum grown in submerged culture were evaluated for antioxidant potential and anti-inflammatory effects. EAF exhibited the highest total phenolic content and the strongest antioxidant activity based on 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays. HF showed dose-dependent inhibition of NO production in LPS-stimulated RAW264.7 cells and NO radical scavenging activity. Gas chromatographic analysis of HF revealed the presence of ethyl linoleate and ergosterol, compounds with known anti-inflammatory properties. In conclusion, the nutritive compositions and significant antioxidant potential and anti-inflammatory effects of mycelia extracts of A. rugosum have the potential to serve as a therapeutic agent or adjuvant in the management of inflammatory disorders.
This study aimed to determine anthocyanins and their antioxidative and cardioprotective properties in defatted dabai parts. Anthocyanins in crude extracts and extract fractions of defatted dabai peel and pericarp were quantified using UHPLC, while their antioxidant capacity and oxidative stress inhibition ability were evaluated by using DPPH and CUPRAC assays as well as linoleic acid oxidation system, hemoglobin oxidation, and PARP-1 inhibition ELISA. Cardioprotective effect of the defatted dabai peel extract was evaluated using hypercholesterolemic-induced New Zealand white rabbits. Six anthocyanins were detected in the defatted dabai peel, with the highest antioxidant capacities and oxidative stress inhibition effect compared to the other part. The defatted dabai peel extract has also inhibited lipid peroxidation (plasma MDA) and elevated cellular antioxidant enzymes (SOD and GPx) in the tested animal model. Major anthocyanin (cyanidin-3-glucoside) and other anthocyanins (pelargonidin-3-glucoside, malvidin-3-glucoside, cyanidin-3-galactoside, cyanidin-3-arabinoside, and peonidin-3-glucoside) detected in the defatted dabai peel are potential future nutraceuticals with promising medicinal properties.