Structural and chemical insights into the prebiotic property of hemicellulosic polysaccharide from Santalum album L.

Hemicellulose was extracted by alkali treatment of de-pectinated cell wall material of Santalum album L. (sandalwood) suspension culture cells. The physicochemical properties and prebiotic activities of a purified major fraction of Hemicellulose-B, termed as HB-I, were investigated. GC analysis of hydrolyzed and derivatized HB-I showed the presence of arabinose (~64 %), galactose (~16 %) and glucose (~16 %) as major monosaccharide units along with minor amount of rhamnose. Methylation and NMR studies on the purified polysaccharide revealed the presence of 6-ß-d-Glcp, ß-d-Galp, 3,5-α-l-Araf, α-l-Araf, 5-α-l-Araf, 2,3-α-l-Araf and, α-l-Rhap residues, from which a proposed structure of repeating units was established. The growth of probiotic Lactobacillus spp. strains L. acidophilus, L. casei, L. plantarum and L. rhamnosus was promoted while that of Escherichia coli was suppressed significantly in presence of HB-I. Our results highlight valorization of sandalwood biomass and explore the role of mixed α, ß-linked heteroglycan as a potential prebiotic molecule thus indicating the possibility of development of low-cost bioprocesses for production of functional food ingredients.

Structural analysis of a water insoluble polysaccharide from pearl millet and evaluating its prebiotic activity.

Epidemiological studies have shown an inverse correlation between dietary intake of prebiotics and the risk of chronic diseases. Pearl millet is a potential economic source to develop a new class of prebiotics in the form of its polysaccharide. In the present study, the chemical structure of a water insoluble homopolysaccharide (PMG), and its prebiotic properties were investigated. The structure of PMG was elucidated on the basis of total hydrolysis, methylation analysis, and 1D/2D NMR (1H, 13C, DEPT-135, HSQC, DQF-COSY, NOESY and ROESY) experiments. The results indicated that PMG was a glucan with an average molecular weight ~ 361 kDa having a backbone of (1 â†’ 3) α-d-glucopyranosyl residues. Hydrolysis of PMG by salivary and pancreatic α amylase was 1.75 % ± 0.34 and 1.99 % ± 0.18 respectively. A positive prebiotic score of PMG with both L. acidophilus and L. brevis (0.446 ± 0.031 & 0.427 ± 0.016) hints towards its prebiotic potential. These observations suggest that PMG might be used as a potential prebiotic component in the food and pharmaceutical applications.

2D Hexagonal Boron Nitride-Coated Cotton Fabric with Self-Extinguishing Property.

Here, we report on the fabrication of flame retardant hydrophobic cotton fabrics based on the coating with two-dimensional hexagonal boron nitride (2D hBN) nanosheets. A simple one-step solution dipping process was used to coat the fabrics by taking advantage of the strong bonding between diethylenetriamine and hBN on the cotton surface. Exposure to direct flame confirmed the improvement of the flame retardant properties of the coated cotton fabrics. In turn, removal of the flame source revealed self-extinguishing properties. Molecular dynamics simulations indicate that hBN hinders combustion by reducing the rate at which oxygen molecules reach the cotton surface. This time-saving and one-step approach for the fabrication of flame-retardant cotton fabrics offers significant advantages over other, less efficient production methods.

Molecular Docking Studies of a Cyclic Octapeptide-Cyclosaplin from Sandalwood.

Natural products from plants, such as chemopreventive agents, attract huge attention because of their low toxicity and high specificity. The rational drug design in combination with structure-based modeling and rapid screening methods offer significant potential for identifying and developing lead anticancer molecules. Thus, the molecular docking method plays an important role in screening a large set of molecules based on their free binding energies and proposes structural hypotheses of how the molecules can inhibit the target. Several peptide-based therapeutics have been developed to combat several health disorders, including cancers, metabolic disorders, heart-related diseases, and infectious diseases. Despite the discovery of hundreds of such therapeutic peptides however, only few peptide-based drugs have made it to the market. Moreover, the in silico activities of cyclic peptides towards molecular targets, such as protein kinases, proteases, and apoptosis related proteins have not been extensively investigated. In this study, we explored the in silico kinase and protease inhibitor potentials of cyclosaplin, and studied the interactions of cyclosaplin with other apoptosis-related proteins. Previously, the structure of cyclosaplin was elucidated by molecular modeling associated with dynamics that were used in the current study as well. Docking studies showed strong affinity of cyclosaplin towards cancer-related proteins. The binding affinity closer to 10 kcal/mol indicated efficient binding. Cyclosaplin showed strong binding affinities towards protein kinases such as EGFR, VEGFR2, PKB, and p38, indicating its potential role in protein kinase inhibition. Moreover, it displayed strong binding affinity to apoptosis-related proteins and revealed the possible role of cyclosaplin in apoptotic cell death. The protein-ligand interactions using LigPlot displayed some similar interactions between cyclosaplin and peptide-based ligands, especially in case of protein kinases and a few apoptosis related proteins. Thus, the in silico analyses gave the insights of cyclosaplin being a potential apoptosis inducer and protein kinase inhibitor.

Evaluation of Cyclosaplin Efficacy Using a Silk Based 3D Tumor Model.

Development of novel anti-cancer peptides requires a rapid screening process which can be accelerated by using appropriate in vitro tumor models. Breast carcinoma tissue is a three-dimensional (3D) microenvironment, which contains a hypoxic center surrounded by dense proliferative tissue. Biochemical clues provided by such a 3D cell mass cannot be recapitulated in conventional 2D culture systems. In this experiment, we evaluate the efficacy of the sandalwood peptide, cyclosaplin, on an established in vitro 3D silk breast cancer model using the invasive MDA-MB-231 cell line. The anti-proliferative effect of the peptide on the 3D silk tumor model is monitored by alamarBlue assay, with conventional 2D culture as control. The proliferation rate, glucose consumed, lactate dehydrogenase (LDH), and matrix metalloproteinase 9 (MMP-9) activity of human breast cancer cells are higher in 3D constructs compared to 2D. A higher concentration of drug is required to achieve 50% cell death in 3D culture than in 2D culture. The cyclosaplin treated MDA-MB-231 cells showed a significant decrease in MMP-9 activity in 3D constructs. Microscopic analysis revealed the formation of cell clusters evenly distributed in the scaffolds. The drug treated cells were less in number, smaller and showed unusual morphology. Overall, these findings indicate the role of cyclosaplin as a promising anti-cancer therapeutic.

One- and two-photon responsive sulfur dioxide (SO2) donors: a combinatorial drug delivery for improved antibiotic therapy.

One- and two-photon activated sulfur dioxide donors based on a 4,5-dimethoxy-2-nitrobenzyl phototrigger have been developed. The designed donors have the ability to release not only SO2 but also a hydroxy-compound in a simultaneous manner. Furthermore, we demonstrated their application in combinatorial therapy by the dual release of SO2 and an active drug, i.e. ferulic acid ethyl ester (FAEE) with self-monitoring ability. Next, we investigated the in vitro cellular uptake and the capability of SO2 generation from the donors using a well-known coumarin-hemicyanine fluorescent probe. Finally, we evaluated the antibacterial activity of the designed donors (5a, 5b and 6) by broth dilution and agar well diffusion methods on E. cloacae cells (MTCC 509). The results show that the donor 5a exhibits enhanced antibacterial efficacy compared to 5b and 6, due to the synergetic effect of dually released SO2 and FAEE.

Sphingobactan, a new α-mannan exopolysaccharide from Arctic Sphingobacterium sp. IITKGP-BTPF3 capable of biological response modification.

An exopolysaccharide, from a new Arctic permafrost isolate, Sphingobacterium sp. IITKGP-BTPF3 was purified and characterized. Upon optimization of various parameters (pH, temperature, carbon and nitrogen source), the yield of EPS obtained was 1.42 g/L. Structural investigation through FT-IR, GC-MS/MS, HPLC and NMR (1D and 2D) revealed the molecule to be a mannan with α-(1 → 2) and α-(1 → 6) linkages. Anti-oxidant and macrophage immunomodulatory assays were employed for the assessment of bioactivity. Sphingobactan was found to be capable of scavenging superoxide anions, and reducing the nitric oxide production in LPS elicited murine macrophage (RAW 264.7) cell line. The in vitro findings indicate the potential of Sphingobactan as a biological response modification (BRM) agent, for containment and possible resolution of inflammatory response in vivo.

Silk fibroin-Thelebolan matrix: A promising chemopreventive scaffold for soft tissue cancer.

Research of improved functional bio-mimetic matrix for regenerative medicine is currently one of the rapidly growing fields in tissue engineering and medical sciences. This study reports a novel bio-polymeric matrix, which is fabricated using silk protein fibroin from Bombyx mori silkworm and fungal exopolysaccharide Thelebolan from Antarctic fungus Thelebolus sp. IITKGP-BT12 by solvent evaporation and freeze drying method. Natural cross linker genipin is used to imprison the Thelebolan within the fibroin network. Different cross-linked and non-cross-linked fibroin/Thelebolan matrices are fabricated and biophysically characterized. Cross-linked thin films show robustness, good mechanical strength and high temperature stability in comparison to non-cross-linked and pure matrices. The 3D sponge matrices demonstrate good cytocompatibility. Interestingly, sustained release of the Thelebolan from the cross-linked matrices induce apoptosis in colon cancer cell line (HT-29) in time dependent manner while it is nontoxic to the normal fibroblast cells (L929).The findings indicate that the cross-linked fibroin/Thelebolan matrices can be used as potential topical chemopreventive scaffold for preclusion of soft tissue carcinoma.

A p-Hydroxyphenacyl-Benzothiazole-Chlorambucil Conjugate as a Real-Time-Monitoring Drug-Delivery System Assisted by Excited-State Intramolecular Proton Transfer.

Among the well-known phototriggers, the p-hydroxyphenacyl (pHP) group has consistently enabled the very fast, efficient, and high-conversion release of active molecules. Despite this unique behavior, the pHP group has been ignored as a delivery agent, particularly in the area of theranostics, because of two major limitations: Its excitation wavelength is below 400 nm, and it is nonfluorescent. We have overcome these limitations by incorporating a 2-(2'-hydroxyphenyl)benzothiazole (HBT) appendage capable of rapid excited-state intramolecular proton transfer (ESIPT). The ESIPT effect also provided two unique advantages: It assisted the deprotonation of the pHP group for faster release, and it was accompanied by a distinct fluorescence color change upon photorelease. In vitro studies showed that the p-hydroxyphenacyl-benzothiazole-chlorambucil conjugate presents excellent properties, such as real-time monitoring, photoregulated drug delivery, and biocompatibility.

Fluorene-morpholine-based organic nanoparticles: lysosome-targeted pH-triggered two-photon photodynamic therapy with fluorescence switch on-off.

Nanocarrier-mediated photodynamic therapy (PDT) is an effective tool for anti-tumour treatment due to the targeted and image-guided delivery of photosensitizers (PSs) to diseased tissues. These nanocarriers range from inorganic, ceramic, polymeric to biological nanoparticles (NPs). Such PS-grafted bicomponent nanocarriers have limitations like (i) difficulty in surface modification, (ii) lower loading percentages of the therapeutic agent, (iii) unstable physical encapsulation, etc. By any means, if we can prepare PSs directly as NPs then we can surpass the above drawbacks. Hence, we synthesised new two-photon fluorene-functionalised morpholine (Fluo-Mor)-based organic NPs that showed strong fluorescence and profound photodynamic therapy (PDT) activity only in acidic medium. Such a pH-responsive appearance of fluorescence enables Fluo-Mor NPs for the real time monitoring of photodynamic therapeutic activity selectively in low-pH organelles viz. lysosome. Cytotoxicity of Fluo-Mor NPs was monitored using time-dependent and dose-dependent cancer cell viability assay and confocal imaging.

Silver nanoparticle anchored carbon dots for improved sensing, catalytic and intriguing antimicrobial activity.

Fluorescent carbon dots (NSCDs) with a size of ∼5 nm (λex = 320 nm and λem = 386 nm) have been synthesized under reflux from an alkaline mixture of dopamine and cysteine. The synthesized NSCDs are hybridized with in situ generated silver nanoparticles (AgNPs) obtained by mixing AgNO3 at room temperature. NSCDs enrich the plasmonic bands of AgNPs due to the localized surface plasmon resonance (LSPR) effect. Further enrichment of plasmon band, depending on the acetone concentration, enables acetone sensing down to 8 × 10(-5) M admixed in 1 M water. Thus, acetone induced hybrid particles with a sharp plasmon band (λex = 410 nm) become a sulfide sensing platform. Furthermore, vacuum dried stable particles (with or without acetone) have been proven to be an excellent catalyst for selective reduction of cationic dyes and they exhibit intriguing antimicrobial activity. These two types of dry particle act differently, which enables us to distinguish their altered surface functionalization in terms of catalysis and bacterial growth.

Coumarin-benzothiazole-chlorambucil (Cou-Benz-Cbl) conjugate: an ESIPT based pH sensitive photoresponsive drug delivery system.

We have developed an ESIPT based drug delivery system (DDS), Cou-Benz-Cbl conjugate, by incorporating a benzothiazole group at the 8th position of the 7-hydroxy-coumarin moiety for pH sensitive fluorescence properties and photocontrolled release of the anticancer drug chlorambucil. The Cou-Benz-Cbl conjugate exhibited unique photophysical properties like good absorbance at around 350 nm, a large Stokes shift (∼151 nm) and pH sensitive fluorescence properties. The pH sensitive fluorescence properties of the Cou-Benz-Cbl conjugate can be ascribed to an ESIPT turn "on and off" mechanism. At physiological pH, the ESIPT gets turned "off" and a blue fluorescence of the coumarin moiety was observed, but at acidic pH, the ESIPT gets turned "on" and a green fluorescence was noted. Photolysis of the Cou-Benz-Cbl conjugate using UV light of wavelength ≥365 nm resulted in the efficient release of the anticancer drug chlorambucil. Cellular uptake studies revealed that the Cou-Benz-Cbl conjugate was easily internalized inside the cancer cells. Further, an MTT assay showed that the Cou-Benz-Cbl conjugate has a good biocompatibility and low cytotoxicity towards the MDA-MB-231 cell line, whereas upon exposure to UV light, the Cou-Benz-Cbl conjugate exhibited enhanced cytotoxicity compared to the free drug due to the effective release of the anticancer drug chlorambucil inside the cancer cell.

Fabrication of superhydrophobic copper surface on various substrates for roll-off, self-cleaning, and water/oil separation.

Superhydrophobic surfaces prevent percolation of water droplets and thus render roll-off, self-cleaning, corrosion protection, etc., which find day-to-day and industrial applications. In this work, we developed a facile, cost-effective, and free-standing method for direct fabrication of copper nanoparticles to engender superhydrophobicity for various flat and irregular surfaces such as glass, transparency sheet (plastic), cotton wool, textile, and silicon substrates. The fabrication of as-prepared superhydrophobic surfaces was accomplished using a simple chemical reduction of copper acetate by hydrazine hydrate at room temperature. The surface morphological studies demonstrate that the as-prepared surfaces are rough and display superhydrophobic character on wetting due to generation of air pockets (The Cassie-Baxter state). Because of the low adhesion of water droplets on the as-prepared surfaces, the surfaces exhibited not only high water contact angle (164 ± 2°, 5 µL droplets) but also superb roll-off and self-cleaning properties. Superhydrophobic copper nanoparticle coated glass surface uniquely withstands water (10 min), mild alkali (5 min in saturated aqueous NaHCO3 of pH ≈ 9), acids (10 s in dilute HNO3, H2SO4 of pH ≈ 5) and thiol (10 s in neat 1-octanethiol) at room temperature (25-35 °C). Again as-prepared surface (cotton wool) was also found to be very effective for water-kerosene separation due to its superhydrophobic and oleophilic character. Additionally, the superhydrophobic copper nanoparticle (deposited on glass surface) was found to exhibit antibacterial activity against both Gram-negative and Gram-positive bacteria.

1-Acetylpyrene-salicylic acid: photoresponsive fluorescent organic nanoparticles for the regulated release of a natural antimicrobial compound, salicylic acid.

Photoresponsive 1-acetylpyrene-salicylic acid (AcPy-SA) nanoparticles (NPs) were developed for the regulated release of a natural antimicrobial compound, salicylic acid. The strong fluorescent properties of AcPy-SA NPs have been extensively used for potential in vitro cell imaging. The phototrigger capability of our newly prepared AcPy-SA NPs was utilized for the efficient release of an antimicrobial compound, salicylic acid. The photoregulated drug release of AcPy-SA NPs has been shown by the subsequent switching off and on of a visible-light source. In vitro biological studies reveal that AcPy-SA NPs of ∼68 nm size deliver the antimicrobial drug salicylic acid into the bacteria cells (Pseudomonas aeruginosa) and efficiently kill the cells upon exposure to visible light (≥410 nm). Such photoresponsive fluorescent organic NPs will be highly beneficial for targeted and regulated antimicrobial drug release because of their biocompatible nature, efficient cellular uptake, and light-induced drug release ability.

Isolation and characterization of extracellular polysaccharide Thelebolan produced by a newly isolated psychrophilic Antarctic fungus Thelebolus.

The present investigation is on a newly isolated psychrophilic Antarctic filamentous Ascomycetous fungus that has been identified as Thelebolus sp. and given the designation of Thelebolus sp. IITKGP-BT12. The culture was primarily identified through morphological studies, and was further confirmed by 18S rRNA sequencing (GenBank Accession No. KC191572), which revealed its close relatedness with Thelebolus microsporus. The exopolysaccharide (EPS) produced (1.94 g L(-1)) by the fungus was isolated, purified and characterized as glucan having an average molecular mass of 5×10(5)Da. The structure of EPS was determined by gas chromatography with tandem mass spectrometry (GC-MS/MS), Fourier transform infrared spectroscopy (FT-IR) and nuclear magnetic resonance (NMR) studies ((1)H, (13)C and HSQC). NMR analysis indicated the presence of (1→3)-linked ß-d-glucan backbone with (1→6)-linked branches of ß-d-glucopyranosyl units. Antiproliferative activity of EPS was demonstrated in B16-F0 cells, with IC50 of 275.42 µg m L(-1). Flow cytometry analysis and DNA fragmentation studies revealed that the cytotoxic action of the EPS mediated apoptosis in cancer cells. This is the first ever report on bioactive EPS thelebolan from Thelebolus sp.

Identification and structural characterization of a new pro-apoptotic cyclic octapeptide cyclosaplin from somatic seedlings of Santalum album L.

Small cyclic peptides exhibiting potent biological activity have great potential for anticancer therapy. An antiproliferative cyclic octapeptide, cyclosaplin was purified from somatic seedlings of Santalum album L. (sandalwood) using gel filtration and RP-HPLC separation process. The molecular mass of purified peptide was found to be 858 Da and the sequence was determined by MALDI-ToF-PSD-MS as 'RLGDGCTR' (cyclic). The cytotoxic activity of the peptide was tested against human breast cancer (MDA-MB-231) cell line in a dose and time-dependent manner. The purified peptide exhibited significant antiproliferative activity with an IC50 2.06 µg/mL. In a mechanistic approach, apoptosis was observed in differential microscopic studies for peptide treated MDA-MB-231 cells, which was further confirmed by mitochondrial membrane potential, DNA fragmentation assay, cell cycle analysis and caspase 3 activities. The modeling and docking experiments revealed strong affinity (kcal/mol) of peptide toward EGFR and procaspase 3. The co-localization studies revealed that the peptide sensitizes MDA-MB-231 cells by possibly binding to EGFR and induces apoptosis. This unique cyclic octapeptide revealed to be a favorable candidate for development of anticancer agents.

Water soluble bioactives of nacre mediate antioxidant activity and osteoblast differentiation.

The water soluble matrix of nacre is a proven osteoinductive material. In spite of the differences in the biomolecular compositions of nacre obtained from multiple species of oysters, the common biochemical properties of those principles substantiate their biological activity. However, the mechanism by which nacre stimulates bone differentiation remains largely unknown. Since the positive impact of antioxidants on bone metabolism is well acknowledged, in this study we investigated the antioxidant potential of a water soluble matrix (WSM) obtained from the nacre of the marine oyster Pinctada fucata, which could regulate its osteoblast differentiation activity. Enhanced levels of ALP activity observed in pre-osteoblast cells upon treatment with WSM, suggested the induction of bone differentiation events. Furthermore, bone nodule formation and up-regulation of bone differentiation marker transcripts, i.e. collagen type-1 and osteocalcin by WSM confirmed its ability to induce differentiation of the pre-osteoblasts into mature osteoblasts. Remarkably, same WSM fraction upon pre-treatment lowered the H2O2 and UV-B induced oxidative damages in keratinocytes, thus indicating the antioxidant potential of WSM. This was further confirmed from the in vitro scavenging of ABTS and DPPH free radicals and inhibition of lipid peroxidation by WSM. Together, these results indicate that WSM poses both antioxidant potential and osteoblast differentiation property. Thus, bioactivities associated with nacre holds potential in the development of therapeutics for bone regeneration and against oxidative stress induced damages in cells.

Purification and characterization of a betanidin glucosyltransferase from Amaranthus tricolor L catalyzing non-specific biotransformation of flavonoids.

Betacyanins are the major pigments present in Amaranthus tricolor, a leafy vegetable consumed globally. The terminal glycosylation of the aglycone betanidin is an important step in the biosynthesis of this natural red antioxidant pigment. A betanidin 5-O-glucosyltransferase (BGT) was fully purified to 134 folds (specific activity, 265.2 nkat mg(-1)) from the red amaranth by ammonium sulfate precipitation followed by hydrophobic interaction, anion exchange and size exclusion chromatography. Homogeneity of the purified protein was confirmed by 2-dimensional polyacrylamide gel electrophoresis (2D PAGE). The molecular weight of the enzyme determined by liquid chromatography-mass spectrometry (LC-MS) was found to be 62.8 kDa. Furthermore, the enzyme glycosylated flavonoids (kaempferol and quercetin) but not anthocyanidins, presence of which is mutually exclusive to betacyanin accumulating plants. The apparent Km (344±2.34 µM) and Vmax (17.24 µM min(-1)) of the enzyme were determined by LC-MS/MS. Peptide mass fingerprinting of the purified protein showed 38.4% coverage of peptide masses with anthocyanidin 3-O-glucosyltransferase from Zea mays. Study on this purified enzyme, for the first time, revealed its role of glycosylation in biosynthesis of betacyanin in A. tricolor and indicates promiscuous substrate-specificity.

TLC-bioautographic evaluation of in vitro anti-tyrosinase and anti-cholinesterase potentials of sandalwood oil.

Sandalwood oil, rich in sesquiterpenoid alcohols, has been used in traditional medicinal systems as a relaxant and coolant. Besides, sandalwood oil is used as an ingredient in numerous skin fairness enhancing cosmetics. However, there is no available information on biological activities that relate to the above applications. Hence, the anti-tyrosinase and anti-cholinesterase potentials of sandalwood oil were probed by both TLC-bioautographic and colorimetric methods. Results obtained from colorimetric assays indicated that sandalwood oil is a potent inhibitor of tyrosinase (IC50 = 171 microg mL(-1)) and cholinesterases (IC50 = 4.8-58 microg mL(-1)), in comparison with the positive controls used in the assays, kojic acid and physostigmine, respectively. The TLC-bioautographic assays indicated that alpha-santalol, the major constituent of the oil, is a strong inhibitor of both tyrosinase and cholinesterase. These in vitro results indicate that there is a great potential of this essential oil for use in the treatment of Alzheimer's disease, as well as in skin-care.

Evaluation of in vivo anti-hyperglycemic and antioxidant potentials of α-santalol and sandalwood oil.

Sandalwood finds numerous mentions across diverse traditional medicinal systems in use worldwide. The objective of this study was to evaluate the in vivo anti-hyperglycemic and antioxidant potential of sandalwood oil and its major constituent α-santalol. The in vivo anti-hyperglycemic experiment was conducted in alloxan-induced diabetic male Swiss albino mice models. The in vivo antioxidant experiment was performed in d-galactose mediated oxidative stress induced male Swiss albino mice models. Intraperitoneal administration of α-santalol (100mg/kg BW) and sandalwood oil (1g/kg BW) for an week modulated parameters such as body weight, blood glucose, serum bilirubin, liver glycogen, and lipid peroxides contents to normoglycemic levels in the alloxan-induced diabetic mice. Similarly, intraperitoneal administration of α-santalol (100mg/kg BW) and sandalwood oil (1g/kg BW) for two weeks modulated parameters such as serum aminotransferases, alkaline phosphatase, bilirubin, superoxide dismutase, catalase, free sulfhydryl, protein carbonyl, nitric oxide, liver lipid peroxide contents, and antioxidant capacity in d-galactose mediated oxidative stress induced mice. Besides, it was observed that the beneficial effects of α-santalol were well complimented, differentially by other constituents present in sandalwood oil, thus indicating synergism in biological activity of this traditionally used bioresource.