Inducing antioxidant and antimicrobial activities in endophytic and endolichenic fungi by the use of light spectra treatments.

The influence of light exposure on antioxidant and antimicrobial activities of nine fungal isolates [Pseudopestalotiopsis theae (EF13), Fusarium solani (EF5), Xylaria venustula (PH22), Fusarium proliferatum (CCH), Colletotrichum boninese (PL9), Colletotrichum boninese (PL1), Colletotrichum boninese (OL2), Colletotrichum gloeosporioides (OL3) and Colletotrichum siamense (PL3)] were determined. The isolates were incubated in blue, red, green, yellow and white fluorescent light (12 h photoperiod of alternating light/dark). It was observed that green light induced higher total phenolic content (TPC) (2.96 ± 0.16 mg-30.71 ± 1.03 mg GAE/g) and ferric reducing antioxidant power (FRAP) in most isolates (4.82 ± 0.04-53.55 ± 4.33 mg GAE/g), whereas red light induced higher total flavonoid content (TFC) levels (1.14 ± 0.08-18.40 ± 1.12 mg QE/g). The crude extracts from most fungal cultures exposed to green and red lights were also notably more potent against the tested pathogens, as larger zones of inhibition (ZOI) (9.00 ± 1.00-38.30 ± 2.90 mm) and lower minimum inhibitory concentration (MIC) (0.0196-1.25 mg/mL) were achieved for antimicrobial effect. This study showed that light treatments are effective strategies in enhancing production of more potent antimicrobial compounds and valuable antioxidants from fungal isolates.

Influence of light regulation on growth and enzyme production in rare endolichenic fungi.

The influence of light regulation on the growth and enzyme production of three endolichenic fungal isolates, i.e. Pseudopestalotiopsis theae (EF13), Fusarium solani (EF5), and Xylaria venustula (PH22), was determined. The isolates were exposed to blue, red, green, yellow, white fluorescent light (12 h light-12 h dark photoperiod) (test), and 24 h dark (control) conditions. Results revealed that the alternating light-dark conditions resulted in the formation of dark rings in most fungal isolates but was absent in PH22. Red light induced sporulation while yellow light elicited higher biomass in all isolates (0.19 ± 0.01 g, 0.07 ± 0.00 g, and 0.11 ± 0.00 g, for EF13, PH22, and EF5, respectively) as compared to incubation in the dark. Results also showed that blue light induced higher amylase activity in PH22 (15.31 ± 0.45 U/mL) and L-asparaginase activity in all isolates (0.45 ± 0.01 U/mL, 0.55 ± 0.39 U/mL, and 0.38 ± 0.01 U/mL, for EF13, PH22, and EF5, respectively) compared to both control conditions. Green light enhanced the production of xylanase (6.57 ± 0.42 U/mL, 10.64 ± 0.12 U/mL, and 7.55 ± 0.56 U/mL for EF13, PH22, and EF5, respectively) and cellulase (6.49 ± 0.48 U/mL, 9.57 ± 0.25 U/mL, and 7.28 ± 0.63 U/mL, for EF13, PH22, and EF5, respectively). In contrast, red light was the least effective light treatment as production of enzymes was the least, with lower levels of amylase, cellulase, xylanase, and L-asparaginase detected. To conclude, all three endolichenic fungi are light-responsive, with fungal growth regulated with the use of red light and yellow light, and manipulation of enzyme production via blue and green light.

Antioxidant, cytotoxic, and antibacterial activities of Clitoria ternatea flower extracts and anthocyanin-rich fraction.

Clitoria ternatea flower is a traditional medicinal herb that has been used as a natural food colourant. As there are limited studies on investigating the bioactivities of the anthocyanin-rich fraction of Clitoria ternatea flower, this study aimed to determine an efficient column chromatography method to obtain the anthocyanin-rich fraction from this flower and characterise its composition, antioxidant, antibacterial, and cytotoxic activities. Amberlite XAD-16 column chromatography was more efficient in enriching the total anthocyanin content (TAC) of the fraction with the highest TAC to total phenolic content (TPC) ratio of 1:6 than that using C18-OPN. A total of 11 ternatin anthocyanins were characterised in the anthocyanin-rich fraction by LC-MS analysis. The antioxidant activity of the anthocyanin-rich fraction was more potent in the chemical-based assay with an IC50 value of 0.86 ± 0.07 mg/mL using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay than cellular antioxidant assay using RAW 264.7 macrophages. In vitro cytotoxicity assay using human embryonic kidney HEK-293 cell line showed the anthocyanin-rich fraction to be more toxic than the crude extracts. The anthocyanin-rich fraction had more potent antibacterial activity than the crude extracts against Bacillus cereus, Bacillus subtilis, and Escherichia coli. The anthocyanin-rich fraction of C. ternatea has the potential to be used and developed as a functional food ingredient or nutraceutical agent.

Antibiofilm properties of Clitoria ternatea flower anthocyanin-rich fraction towards Pseudomonas aeruginosa.

In Asia, Clitoria ternatea flowers are commonly used as a traditional medicinal herb and as a food colourant. Their bioactive compounds have anti-inflammatory, anti-microbial and anti-biofilm activities. Pseudomonas aeruginosa is one of the major pathogens that cause biofilm-associated infections resulting in an increase in antimicrobial resistance. Hence, the aim of this study was to investigate if the anti-biofilm properties of the anthocyanin-rich fraction of C. ternatea flowers were effective against P. aeruginosa . The effect of the anthocyanin-rich fraction of C. ternatea flowers on P. aeruginosa biofilms formed on a polystyrene surface was determined using the crystal violet assay and scanning electron microscopy (SEM). The anthocyanin-rich fraction reduced biofilm formation by four P. aeruginosa strains with a minimum biofilm inhibitory concentration value ranging between 0.625 and 5.0 mg ml-1. We further show that the biofilm-inhibiting activity of C. ternatea flowers is not due to the flavonols but is instead attributed to the anthocyanins, which had significant biofilm inhibitory activity (64.0±1.1 %) at 24 h in a time-response study. The anthocyanin-rich fraction also significantly reduced bacterial attachment on the polystyrene by 1.1 log c.f.u. cm-2 surface based on SEM analysis. Hence, anthocyanins from C. ternatea flowers have potential as an agent to decrease the risk of biofilm-associated infections.

Removal of triphenylmethane dyes by Streptomyces bacillaris: A study on decolorization, enzymatic reactions and toxicity of treated dye solutions.

This study revealed Streptomyces bacillaris as an efficient biological agent for the removal of triphenylmethane (TPM) dyes. The isolate decolorized Malachite Green (MG), Methyl Violet (MV), Crystal Violet (CV), and Cotton Blue (CB) effectively. S. bacillaris in the treated dye solutions were analyzed for enzyme production, and the cell biomass was observed for functional groups and cell morphology. The treated dye solutions were also analyzed for degraded compounds and their toxicity. Results revealed high decolorization activities for MG (94.7%), MV (91.8%), CV (86.6%), CB (68.4%), attributed to both biosorption and biodegradation. In biosorption, dye molecules interacted with the hydroxyl, amino, phosphoryl, and sulfonyl groups present on the cell surface. Biodegradation was associated with induced activities of MnP and NADH-DCIP reductase, giving rise to various simpler compounds. The degraded compounds in the treated dyes were less toxic, as revealed by the significant growth of Vigna radiata in the phytotoxicity test. There were no significant changes in cell morphology before and after use in dye solutions, suggesting S. bacillaris is less susceptible to dye toxicity. This study concluded that S. bacillaris demonstrated effective removal of TPM dyes via biosorption and biodegradation, rendering the treated dyes less toxic than untreated dyes. Findings in this study enabled further explorations into the potential application of lesser-known actinobacteria (i.e. Streptomyces sp.) for dye removal.

Sources and relative stabilities of acylated and nonacylated anthocyanins in beverage systems.

Anthocyanins are considered as the largest group of water-soluble pigments found in the vacuole of plant cells, displaying range of colors from pink, orange, red, purple and blue. They belong to flavonoids, a polyphenolic subgroup. Application of anthocyanins in food systems as natural food colourants is limited due to the lack of stability under different environmental conditions such as light, pH, heat etc. Anthocyanins esterified with one or more acid groups are referred as acylated anthocyanins. Based on the presence or absence of acyl group, anthocyanins are categorized as acylated and nonacylated anthocyanins. Acylated anthocyanins are further classified as mono, di, tri, tetra acylated anthocyanins according to the number of acyl groups present in the anthocyanin. This review classifies common anthocyanin sources into non-acylated, mono-, di-, tri- and tetra-acylated anthocyanins based on the major anthocyanins present in these sources. The relative stabilities of these anthocyanins with respect to thermal, pH and photo stress in beverage systems are specifically discussed. Common anthocyanin sources such as elderberry, blackberry, and blackcurrant mainly contain nonacylated anthocyanins. Red radish, purple corn, black carrot also mainly contain mono acylated anthocyanins. Red cabbage and purple sweet potato have both mono and diacylated anthocyanins. Poly acylated anthocyanins show relatively higher stability compared with nonacylated and monoacylated anthocyanins. Several techniques such as addition of sweeteners, co-pigmentation and acylation techniques could enhance the stability of nonacylated anthocyanins. Flowers are main sources of polyacylated anthocyanins having higher stability, yet they have not been commercially exploited for their anthocyanins.

Dietary phytochemicals that influence gut microbiota: Roles and actions as anti-Alzheimer agents.

The last decide has witnessed a growing research interest in the role of dietary phytochemicals in influencing the gut microbiota. On the other hand, recent evidence reveals that dietary phytochemicals exhibit properties of preventing and tackling symptoms of Alzheimer's disease, which is a neurodegenerative disease that has also been linked with the status of the gut microbiota over the last decade. Till now, little serious discussions, however, have been made to link recent understanding of Alzheimer's disease, dietary phytochemicals and the gut microbiota together and to review the roles played by phytochemicals in gut dysbiosis induced pathologies of Alzheimer's disease. Deciphering these connections can provide insights into the development and future use of dietary phytochemicals as anti-Alzheimer drug candidates. This review aims at presenting latest evidence in the modulating role of phytochemicals in the gut microbiota and its relevance to Alzheimer's disease and summarizing the mechanisms behind the modulative activities. Limitations of current research in this field and potential directions will also be discussed for future research on dietary phytochemicals as anti-Alzheimer agents.

Extraction methods of butterfly pea (Clitoria ternatea) flower and biological activities of its phytochemicals.

Clitoria ternatea or commonly known as 'Butterfly pea' has been used traditionally in Ayurvedic medicine in which various parts of the plants are used to treat health issues such as indigestion, constipation, arthritis, skin diseases, liver and intestinal problems. The flowers of C. ternatea are used worldwide as ornamental flowers and traditionally used as a food colorant. This paper reviews the recent advances in the extraction and biological activities of phytochemicals from C. ternatea flowers. The application of maceration or ultrasound assisted extraction greatly increased the yield (16-247% of increase) of phytochemicals from C. ternatea flowers. Various phytochemicals such as kaempferol, quercetin and myricetin glycosides as well as anthocyanins have been isolated from C. ternatea flowers. Clitoria ternatea flower extracts were found to possess antimicrobial, antioxidant, anti-inflammatory, cytotoxic and antidiabetic activities which are beneficial to human health. Clitoria ternatea flower is a promising candidate for functional food applications owing to its wide range of pharmacotherapeutic properties as well as its safety and effectiveness.

Choices of chromatographic methods as stability indicating assays for pharmaceutical products: A review.

Stability indicating assay describes a technique which is used to analyse the stability of drug substance or active pharmaceutical ingredient (API) in bulk drug and pharmaceutical products. Stability indicating assay must be properly validated as per ICH guidelines. The important components in a stability indicating assay include sensitivity, specificity, accuracy, reliability, reproducibility and robustness. A validated assay is able to measure the concentration changes of drug substance/API with time and make reliable estimation of the quantity of the degradation impurities. The drug substance is separated and resolved from the impurities. Pros and cons of HPLC, GC, HPTLC, CE and SFC were discussed and reviewed. Stability indicating assay may consist of the combination of chromatographic separation and spectroscopic detection techniques. Hyphenated system could demonstrate parallel quantitative and qualitative analysis of drug substances and impurities. Examples are HPLC-DAD, HPLC-FL, GC-MS, LC-MS and LC-NMR. The analytes in the samples are separated in the chromatography while the impurities are chemically characterised by the spectroscopy in the system. In this review, various chromatographic methods which had been employed as stability indicating assays for drug substance and pharmaceutical formulation were systematically reviewed, and the application of hyphenated techniques in impurities characterisation and identification were also discussed with supporting literatures.

Anthocyanins From Clitoria ternatea Flower: Biosynthesis, Extraction, Stability, Antioxidant Activity, and Applications.

Clitoria ternatea plant is commonly grown as an ornamental plant and possesses great medicinal value. Its flower is edible and also known as blue pea or butterfly pea flower. The unique feature of anthocyanins present in blue pea flowers is the high abundance of polyacylated anthocyanins known as ternatins. Ternatins are polyacylated derivatives of delphinidin 3,3',5'-triglucoside. This review covers the biosynthesis, extraction, stability, antioxidant activity, and applications of anthocyanins from Clitoria ternatea flower. Hot water extraction of dried or fresh petals of blue pea flower could be employed successfully to extract anthocyanins from blue pea flower for food application. Blue pea flower anthocyanins showed good thermal and storage stability, but less photostability. Blue pea flower anthocyanins also showed an intense blue colour in acidic pH between pH 3.2 to pH 5.2. Blue pea flower anthocyanin extracts demonstrate significant in vitro and cellular antioxidant activities. Blue pea flower anthocyanins could be used as a blue food colourant in acidic and neutral foods. The incorporation of blue pea flower anthocyanins in food increased the functional properties of food such as antioxidant and antimicrobial properties. Blue pea flower anthocyanins have also been used in intelligent packaging. A comparison of blue pea flower anthocyanins with two other natural blue colouring agents used in the food industry, spirulina or phycocyanin and genipin-derived pigments is also covered. Anthocyanins from blue pea flowers are promising natural blue food colouring agent.

Anti-dengue virus serotype 2 activity of tannins from porcupine dates.

BACKGROUND: Dengue fever is currently endemic in tropical and subtropical countries worldwide and effective drug against DENV infection is still unavailable. Porcupine dates, which are traditionally used to treat dengue fever, might contain potential anti-dengue compounds. Two porcupine dates, black date (BD) and powdery date (PD) from Himalayan porcupine (Hystrix brachyura), were investigated for their antiviral activities against DENV-2 in vitro. METHODS: The methanol crude extracts (MBD and MPD) were prepared from the raw material of porcupine dates. The tannin-rich fractions (BDTF and PDTF) were isolated from their methanol crude extracts using column chromatography. The presence of tannins in BDTF and PDTF extracts was determined by fourier-transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR) analyses. The cytotoxicity and anti-DENV-2 activities including virus yield inhibition, virucidal, virus attachment and pre-treatment assays of the extracts were examined in Vero cells. RESULTS: Our findings revealed that all the extracts of porcupine dates exhibited antiviral activity against DENV-2 in Vero cells. The IC50 of BDTF and PDTF were 25 µg/mL and 11 µg/mL respectively, while their methanol crude extracts demonstrated lower antiviral efficacy (IC50 ≈ 101-107 µg/mL). BDTF and PDTF also exerted a similar higher virucidal effect (IC50 of 11 µg/mL) than methanol crude extracts (IC50 ≈ 52-66 µg/mL). Furthermore, all the extracts inhibited the attachment of DENV-2 by at least 80%. Pre-treatments of cells with BDTF and PDTF markedly prevented DENV-2 infection when compared to methanol crude extracts. CONCLUSION: This study suggests that porcupine dates possess antiviral properties against DENV-2, which is attributed to its tannin compounds.

Electrospun cellulose acetate butyrate/polyethylene glycol (CAB/PEG) composite nanofibers: A potential scaffold for tissue engineering.

Electrospinning is a common method to prepare nanofiber scaffolds for tissue engineering. One of the common cellulose esters, cellulose acetate butyrate (CAB), has been electrospun into nanofibers and studied. However, the intrinsic hydrophobicity of CAB limits its application in tissue engineering as it retards cell adhesion. In this study, the properties of CAB nanofibers were improved by fabricating the composite nanofibers made of CAB and hydrophilic polyethylene glycol (PEG). Different ratios of CAB to PEG were tested and only the ratio of 2:1 resulted in smooth and bead-free nanofibers. The tensile test results show that CAB/PEG composite nanofibers have 2-fold higher tensile strength than pure CAB nanofibers. The hydrophobicity of the composite nanofibers was also reduced based on the water contact angle analysis. As the hydrophilicity increases, the swelling ability of the composite nanofiber increases by 2-fold with more rapid biodegradation. The biocompatibility of the nanofibers was tested with normal human dermal fibroblasts (NHDF). The cell viability assay results revealed that the nanofibers are non-toxic. In addition to that, CAB/PEG nanofibers have better cell attachment compared to pure CAB nanofibers. Based on this study, CAB/PEG composite nanofibers could potentially be used as a nanofiber scaffold for applications in tissue engineering.

Approach to preserve phenolics in Thunbergia laurifolia leaves by different drying treatments.

Different drying methods and blanching were investigated as to their effects on antioxidant and oxidase activities of Thunbergia laurifolia leaves. Results showed that oven-drying had the highest degradation of total phenolic content (TPC) and antioxidant activity at >85%, while freeze-drying had the lowest at <20%. However, inactivation of oxidase enzymes by blanching at 100°C resulted in a lesser decrease in TPC for oven-drying at 50 and 100°C (51% and 65%, respectively), indicating the importance of inactivating the oxidase enzymes for lower degradation of phenolics on drying. The high-performance liquid chromatography analysis showed that its major antioxidant, rosmarinic acid, degraded tremendously in the presence of oxidase enzymes, but only degraded slightly upon inactivation of oxidase enzymes. Hence, this work showed that by controlling the enzymatic activity, the preservation of phenolics with specific bioactivity in herbal tea leaves can be achieved. PRACTICAL APPLICATIONS: Thunbergia laurifolia leaves have been frequently consumed in the form of a tea or pill due to its medicinal properties. Processing of fresh herbal plant leaves by drying is required to preserve antioxidant phenolic compounds and quality of the plant leaves. Although the drying effects on the antioxidant properties have been studied, the factors that cause the change in properties have not been investigated in-depth. Controlling the factors that affect the phenolic content can help to preserve the beneficial antioxidants when processing the leaves by drying. The result of this study will be of relevance and beneficial to the herbal tea industry.

Current anti-biofilm strategies and potential of antioxidants in biofilm control.

INTRODUCTION: Biofilm formation is a strategy for microorganisms to adapt and survive in hostile environments. Microorganisms that are able to produce biofilms are currently recognized as a threat to human health. Areas covered: Many strategies have been employed to eradicate biofilms, but several drawbacks from these methods had subsequently raised concerns on the need for alternative approaches to effectively prevent biofilm formation. One of the main mechanisms that drives a microorganism to transit from a planktonic to a biofilm-sessile state, is oxidative stress. Chemical agents that could target oxidative stress regulators, for instance antioxidants, could therefore be used to treat biofilm-associated infections. Expert commentary: The focus of this review is to summarize the function and limitation of the current anti-biofilm strategies and will propose the use of antioxidants as an alternative method to treat, prevent and eradicate biofilms. Studies have shown that water-soluble and lipid-soluble antioxidants can reduce and prevent biofilm formation, by influencing the expression of genes associated with oxidative stress. Further in vivo work should be conducted to ensure the efficacy of these antioxidants in a biological environment. Nevertheless, antioxidants are promising anti-biofilm agents, and thus is a potential solution for biofilm-associated infections in the future.

Recycling of phenolic compounds in Borneo's tropical peat swamp forests.

BACKGROUND: Tropical peat swamp forests (TPSF) are globally significant carbon stores, sequestering carbon mainly as phenolic polymers and phenolic compounds (particularly as lignin and its derivatives) in peat layers, in plants, and in the acidic blackwaters. Previous studies show that TPSF plants have particularly high levels of phenolic compounds which inhibit the decomposition of organic matter and thus promote peat accumulation. The studies of phenolic compounds are thus crucial to further understand how TPSF function with respect to carbon sequestration. Here we present a study of cycling of phenolic compounds in five forests in Borneo differing in flooding and acidity, leaching of phenolic compounds from senescent Macaranga pruinosa leaves, and absorption of phenolics by M. pruinosa seedlings. RESULTS: The results of the study show that total phenolic content (TPC) in soil and leaves of three species of Macaranga were highest in TPSF followed by freshwater swamp forest and flooded limestone forest, then dry land sites. Highest TPC values were associated with acidity (in TPSF) and waterlogging (in flooded forests). Moreover, phenolic compounds are rapidly leached from fallen senescent leaves, and could be reabsorbed by tree roots and converted into more complex phenolics within the leaves. CONCLUSIONS: Extreme conditions-waterlogging and acidity-may facilitate uptake and synthesis of protective phenolic compounds which are essential for impeded decomposition of organic matter in TPSF. Conversely, the ongoing drainage and degradation of TPSF, particularly for conversion to oil palm plantations, reverses the conditions necessary for peat accretion and carbon sequestration.

Antioxidant and Intracellular Reactive Oxygen Species/Reactive Nitrogen Species Scavenging Activities of Three Porcupine Bezoars from Hystrix brachyura.

BACKGROUND: Porcupine dates are phytobezoar stones that are used in Traditional Chinese Medicine (TCM) treatments against cancer, postsurgical recovery, dengue fever, etc. The medicinal values have not been scientifically investigated due to the availability and high pricing of the dates. OBJECTIVES: This paper represents the first report on the phytochemical content, in vitro antioxidant and intracellular reactive oxygen species (ROS)/reactive nitrogen species (RNS) scavenging properties of the extracts of three porcupine dates: grassy date (GD), black date (BD), and powdery date (PD). MATERIALS AND METHODS: Dried samples were extracted with methanol and lyophilized. Samples were screened for phytochemical constituents, in vitro antioxidant assays based on total phenolic content (TPC), free radical scavenging, and ferric reducing power (FRP) as well as intracellular ROS and RNS scavenging properties. RESULTS: Phytochemical screening and total tannins assay revealed that tannins, cardiac glycosides, and terpenoids were found in all porcupine dates with tannins forming the major portion of the TPC. In comparison to GD, BD and PD were found to contain significantly high TPC, radical scavenging activity, and FRP. At 200 µg/ml, BD and PD remarkably scavenged 2, 2-azobis (2-amidinopropane) dihydrochloride-induced ROS in RAW264.7 cells and significantly reduced nitric oxide in lipopolysaccharide-stimulated cells. CONCLUSION: Overall, BD and PD exhibited promising in vitro antioxidant as well as intracellular ROS/RNS scavenging properties. SUMMARY: Tannins, cardiac glycoside, and terpenoids were found in all three types of porcupine dates with tannins being the major compoundsAntioxidant contents and properties of three dates were in the order black date (BD) > powdery date (PD) > grassy dateBD and PD extracts showed significant intracellular reactive oxygen species and reactive nitrogen species scavenging properties. Abbreviations Used: TCM: Traditional Chinese Medicine, BD: Black date, GD: Grassy date, PD: Powdery date, TPC: Total phenolic content, FRS: Free radical scavenging, FRP: Ferric reducing power, NO: Nitric oxide, ROS: Reactive oxygen species, RNS: Reactive nitrogen species, GAE: Gallic acid equivalent, AAE: Ascorbic acid equivalent, PVPP: Polyvinylpolypyrrolidone, DCFH-DA: Dichloro-dihydro-fluorescein diacetate, AAPH: 2, 2-azobis (2-amidinopropane) dihydrochloride, LPS: Lipopolysaccharide.

Isolation and Characterisation of a Proanthocyanidin With Antioxidative, Antibacterial and Anti-Cancer Properties from Fern Blechnum orientale.

BACKGROUND: Blechnum orientale Linn. (Blechnaceae), a fern, is traditionally used in the treatment of various ailments, such as skin diseases, stomach pain, urinary bladder complaints, and also as a female contraceptive. Previously, we reported a strong radical scavenging activity, antibacterial activity and cytotoxicity against HT29 colon cancer cells by aqueous extract of B. orientale. OBJECTIVE: In this study, we attempted to isolate and identify the active compound from the aqueous extract of B. orientale. MATERIALS AND METHODS: Aqueous extract of B. orientale was subjected to repeated MCI gel chromatography, Sephadex-LH-20, Chromatorex C18 and semi-preparative high performance liquid chromatography and was characterized using nuclear magnetic resonance and electrospray ionization mass-spectrometry spectroscopic methods. Antioxidant activity was determined using 2, 2-diphenyl-1-picrylhydrazyl radical scavenging assay. Antibacterial assays were conducted using disc diffusion whereas the minimum inhibitory concentration (MIC) and minimum bactericidal concentration were determined using the broth microdilution assay. Cytotoxicity was assessed using thiazolylblue tetrazoliumbromide. RESULTS: A polymeric proanthocyanidin consisting of 2-12 epicatechin extension units and epigallocathecin terminal units linked at C4-C8 was elucidated. Bioactivity studies showed strong radical scavenging activity (IC50 = 5.6 ± 0.1 µg/mL), antibacterial activity (MIC = 31.3-62.5 µg/mL) against five gram-positive bacteria and selective cytotoxicity against HT29 colon cancer cells (IC50 = 7.0 ± 0.3 µg/mL). CONCLUSION: According to our results, the proanthocyanidin of B. orientale demonstrated its potential as a natural source of antioxidant with antibacterial and anti-cancer properties. SUMMARY: A bioactive proanthocyanidin was isolated from the aqueous extract of medicinal fern Blechnum orientale Linn and the structure was elucidated using NMR and ESI-MS spectral studies.The proanthocyanidin compound possessed strong radical scavenging activity (IC50 5.6 ± 0.1 µg/mL)The proanthocyaniding compound showed bactericidal activity against five gram-positive bacteria inclusive of MRSA (minimum inhibitory concentration, MIC and minimum bactericidal concentration, MBC 31.3-62.5 µg/mL).The proanthocyanidin compound is strongly cytotoxic towards cancer cells HT29 (IC50 7.0 ± 0.3 µg/mL), HepG2 (IC50 16 µg/mL) and HCT116 (IC50 20 µg/mL) while weakly cytotoxic towards the non-malignant Chang cells (IC50 48 µg/mL). Abbreviation used: CC: Column chromatography, DP: degree of polymerization, DPPH: 2,2-diphenyl-1-picrylhydrazyl, ESI-MS: electronsprayionisation mass-spectrometry, MBC: Minimum bactericidal concentration, MIC: Minimum inhibitory concentration, MTT: Thiazolyl Blue Tetrazolium Bromide, MRSA: methicillin-resistant Staphylococcus aureus, NMR: nuclear magnetic resonance, TLC: thin layer chromatography, PD: prodelphinidin.

Enzymatic synthesis of quercetin oleate esters using Candida antarctica lipase B.

OBJECTIVES: To investigate the lipase-catalyzed acylation of quercetin with oleic acid using Candida antarctica lipase B. RESULTS: Three acylated analogues were produced: quercetin 4'-oleate (C33H42O8), quercetin 3',4'-dioleate (C51H74O9) and quercetin 7,3',4'-trioleate (C69H106O10). Their identities were confirmed with UPLC-ESI-MS and 1H NMR analyses. The effects of temperature, duration and molar ratio of substrates on the bioconversion yields varied across conditions. The regioselectivity of the acylated quercetin analogues was affected by the molar ratio of substrates. TLC showed the acylated analogues had higher lipophilicity (152% increase) compared to quercetin. Partition coefficient (log P) of quercetin 4'-oleate was higher than those of quercetin and oleic acid. Quercetin 4'-oleate was also stable over 28 days of storage. CONCLUSIONS: Quercetin oleate esters with enhanced lipophilicity can be produced via lipase-catalyzed reaction using C. antarctica lipase B to be used in topical applications.

Antioxidant and tyrosinase inhibition activity of the fertile fronds and rhizomes of three different Drynaria species.

BACKGROUND: For generations, the rhizomes of Drynaria ferns have been used as traditional medicine in Asia. Despite this, the bioactivities of Drynaria rhizomes and leaves have rarely been studied scientifically. METHODS: This study evaluates the antioxidant properties of the methanolic extracts of the fertile fronds and rhizomes from three species in this genus: Drynaria quercifolia, Drynaria rigidula and Drynaria sparsisora. The phenolic and flavonoid contents of the samples were respectively quantified with the total phenolic content (TPC) and total flavonoid content (TFC) assays, while the antioxidant activities were determined via measuring the DPPH radical scavenging activity (FRS), ferric reducing power (FRP), ferrous ion chelating (FIC) activity and lipid peroxidation inhibition (LPI). The tyrosinase inhibition activity of all three species was also reported. RESULTS: The fertile fronds of D. quercifolia were found to exhibit the highest overall TPC (2939 ± 469 mg GAE/100 g) and antioxidant activity amongst all the samples, and the fertile fronds of D. quercifolia and D. rigidula exhibited superior TPC and FRP compared to their rhizomes, despite only the latter being widely used in traditional medicine. The fronds of D. quercifolia had high tyrosinase inhibition activity (56.6 ± 5.0 %), but most of the Drynaria extracts showed unexpected tyrosinase enhancement instead, particularly for D. sparsisora's fronds. CONCLUSION: The high bioactivity of the fertile fronds in the fern species indicate that there is value in further research on the fronds of ferns which are commonly used mostly, or only, for their rhizomes.

Antioxidant and antibacterial activity of Rhoeo spathacea (Swartz) Stearn leaves.

The decoction and infusion of Rhoeo spathacea (Swartz) Stearn leaves have been recognized as a functional food particularly in South America, but has not yet gained international popularity as a beverage. The primary aim of this study was to establish the viability of R. spathacea aqueous leaf extracts as a beverage, in terms of its antioxidant activity and antibacterial activity. The antioxidant contents of aqueous and methanol leaf extracts were evaluated by the total phenolic content (TPC) and total flavonoid content (TFC) assays. The antioxidant activities measured were DPPH radical scavenging activity (FRS), ferric reducing power (FRP) and ferrous ion chelating (FIC) activity. The aqueous leaf extracts in the forms of decoction and infusion, were found to have comparable TPC and antioxidant activity with other herbal teas previously reported by our research group. Both decoction and infusion also exhibited antibacterial activity against six species of Gram positive and four species of Gram negative bacteria, notably methicillin-resistant Staphylococcus aureus and Neisseria gonorrhoeae. A total of four different known phenolic compounds were identified by HPLC and MS, three of which have not been previously reported to be found in this plant. Both the decoction and infusion of the leaves R. spathacea have potential to be popularized into a common beverage.