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  1. Chemical composition of the tiger's milk mushroom, Lignosus rhinocerotis (Cooke) Ryvarden, from different developmental stages
    Lau BF, Abdullah N, Aminudin N
    J Agric Food Chem, 2013 May 22;61(20):4890-7.
    PMID: 23597270 DOI: 10.1021/jf4002507
    The chemical composition of the tiger's milk mushroom (Lignosus rhinocerotis) from different developmental stages, i.e., the fruit body, sclerotium, and mycelium, was investigated for the first time. The fruit body and sclerotium of L. rhinocerotis were rich in carbohydrates and dietary fibers but low in fat. Protein levels in L. rhinocerotis were moderate, and all essential amino acids, except tryptophan, were present. The mycelium contained high levels of potassium, phosphorus, magnesium, riboflavin, and niacin and appreciable amounts of essential fatty acids. The results indicated that the sclerotium of L. rhinocerotis that was used in ethnomedicine was not superior to the fruit body and mycelium with regard to the nutritional content and bioactive constituents. Our findings provide some insights into the selection of appropriate mushroom part(s) of L. rhinocerotis and proper cultivation techniques for the development of new nutraceuticals or dietary supplements.
  2. Comparative SELDI-TOF-MS profiling of low-molecular-mass proteins from Lignosus rhinocerus (Cooke) Ryvarden grown under stirred and static conditions of liquid fermentation
    Lau BF, Aminudin N, Abdullah N
    J Microbiol Methods, 2011 Oct;87(1):56-63.
    PMID: 21801760 DOI: 10.1016/j.mimet.2011.07.005
    Mushrooms are considered as important source of biologically active compounds which include low-molecular-mass protein/peptides (LMMP). In this study, we attempted to profile the LMMP from Lignosus rhinocerus, a wild medicinal mushroom, grown by static cultures (SC) and in stirred tank reactor (STR). Crude water extract (CWE) and protein fractions were profiled using H50 ProteinChip® arrays and SELDI-TOF-MS. Three protein peaks of 5.8, 6.9 and 9.1 kDa were found to be common to spectra of L. rhinocerus CWE from both culture conditions. Partial protein purification has resulted in detection of more peaks in the spectra of protein fractions. For protein fractions of L. rhinocerus cultured in STR, most peaks were observed in the range of 3-8 kDa whereas some peaks with molecular mass up to 14.3 kDa were noted in spectra of protein fractions from SC. Our results have demonstrated the optimization of profiling method using SELDI-TOF-MS for fungal LMMP.
  3. Chemical composition and cellular toxicity of ethnobotanical-based hot and cold aqueous preparations of the tiger's milk mushroom (Lignosus rhinocerotis)
    Lau BF, Abdullah N, Aminudin N, Lee HB
    J Ethnopharmacol, 2013 Oct 28;150(1):252-62.
    PMID: 23993912 DOI: 10.1016/j.jep.2013.08.034
    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.
  4. Ethnomedicinal uses, pharmacological activities, and cultivation of Lignosus spp. (tiger׳s milk mushrooms) in Malaysia - A review
    Lau BF, Abdullah N, Aminudin N, Lee HB, Tan PJ
    J Ethnopharmacol, 2015 Jul 1;169:441-58.
    PMID: 25937256 DOI: 10.1016/j.jep.2015.04.042
    Several members of the genus Lignosus, which are collectively known as cendawan susu rimau (in Malay) or tiger׳s milk mushrooms (TMM), are regarded as important local medicine particularly by the indigenous communities in Malaysia. The mushroom sclerotia are purportedly effective in treating cancer, coughs, asthma, fever, and other ailments. The most commonly encountered Lignosus spp. in Malaysia was authenticated as Lignosus rhinocerotis (Cooke) Ryvarden (synonym: Polyporus rhinocerus), which is also known as hurulingzhi in China and has been used by Chinese physicians to treat liver cancer, gastric ulcers, and chronic hepatitis. In spite of growing interest in the therapeutic potential of TMM, there is no compilation of scientific evidence that supports the ethnomedicinal uses of these mushrooms. Therefore, the present review is intended (i) to provide a comprehensive, up-to-date overview of the ethnomedicinal uses, pharmacological activities, and cultivation of TMM in general and L. rhinocerotis in particular, (ii) to demonstrate how recent scientific findings have validated some of their traditional uses, and (iii) to identify opportunities for future research and areas to prioritize for TMM bioprospecting.
  5. Differential toxicity and teratogenic effects of the hot water and cold water extracts of Lignosusrhinocerus (Cooke) Ryvarden sclerotium on zebrafish (Danio rerio) embryos
    Abd Rashid NA, Lau BF, Kue CS
    J Ethnopharmacol, 2022 Mar 01;285:114787.
    PMID: 34756971 DOI: 10.1016/j.jep.2021.114787
    ETHNOPHARMACOLOGICAL RELEVANCE: The sclerotium of Lignosusrhinocerus (Cooke) Ryvarden is highly valued for its purported medicinal properties. The decoction and macerated materials prepared from the sclerotium are used for treating cancer and other ailments based on extensive traditional knowledge. Scientific evidence from in vitro cytototoxicity, anti-inflammatory and immunomodulatory analyses showed the effectiveness of sclerotial water extracts but toxicity assessment of such preparations has not been reported.

    AIM OF THE STUDY: This study aimed to compare the differential toxicity and teratogenicity (if any) of the hot water (HW) and cold water (CW) extracts of both wild and cultivated sclerotium on zebrafish (Danio rerio) embryos.

    MATERIALS AND METHODS: Zebrafish embryos were treated with varying concentrations of the sclerotial HW and CW extracts (0.3-500 μg/mL) for 72 h until hatching. The hatching, mortality and heartbeat rate of the embryos as well as the potential teratogenic effect of the extracts were assessed in embryos post-treatment with the extracts.

    RESULTS: While the sclerotial HW extracts were nontoxic (LC50 > 500 μg/mL), the sclerotial CW extracts delayed the hatching of the embryos up to 48 h and showed slight toxicity with LC50 values of 398.4 μg/mL and 428.3 μg/mL for the cultivated and wild sclerotium, respectively. The sclerotial CW extracts also induced minor tachycardia in zebrafish larvae. Phenotypic assessment revealed that, while yolk sac edema was observed at high concentrations (300 and 500 μg/mL) of all extracts, curved trunk and bent tail were only observed in the embryos treated with CW extracts of wild sclerotium (300 and 500 μg/mL) but not for CW extracts of cultivated sclerotium at similar concentrations.

    CONCLUSION: The sclerotial water extracts of L.rhinocerus prepared using different methods have varying degree of toxicity and teratogenicity in zebrafish embryos with the sclerotial CW extracts showed higher toxicity than the HW extracts.

  6. Mechanical, durability, and microstructural properties of mortars containing spent mushroom substrate as partial fine aggregate replacement
    Loganathan L, Yap SP, Lau BF, Nagapan M
    Environ Sci Pollut Res Int, 2023 Jun;30(26):69176-69191.
    PMID: 37133663 DOI: 10.1007/s11356-023-27256-y
    Replacing conventional fine aggregates with spent mushroom substrate (SMS) is aimed at developing a sustainable lightweight masonry mortar. It is also an alternative solution for the current improper mushroom waste disposals. Density, workability, compressive strength, specific strength, flexural strength, ultrasonic pulse velocity, water absorption, sorptivity, and equivalent CO2 emission in relation to sand reduction in mortars containing 2.5-15.0% (by volume) SMS passing through a 4.75-mm sieve were investigated. As the percentages of replacement increased from 2.5 to 15.0%, the density of the SMS mortar reduced up to 34.8%, with corresponding compressive strengths of 24.96 to 3.37 MPa. Mixes with up to 12.5% SMS met the minimum compressive and flexural strengths as stated in the ASTM C129 standard. In addition, the equivalent CO2 emission of the mixes reduced 15.09% as the SMS content increased while cost-effectiveness increases up to 98.15% until 7.5% SMS replacement. In conclusion, the use of SMS as fine aggregates up to 12.5% is a viable mix design strategy for producing sustainable lightweight mortar with a lower carbon emission.
  7. Fulltext Evaluation of Selected Culinary-Medicinal Mushrooms for Antioxidant and ACE Inhibitory Activities
    Abdullah N, Ismail SM, Aminudin N, Shuib AS, Lau BF
    Evid Based Complement Alternat Med, 2012;2012:464238.
    PMID: 21716693 DOI: 10.1155/2012/464238
    Considering the importance of diet in prevention of oxidative stress-related diseases including hypertension, this study was undertaken to evaluate the in vitro antioxidant and ACE inhibitory activities of selected culinary-medicinal mushrooms extracted by boiling in water for 30 min. Antioxidant capacity was measured using the following assays: DPPH free radical scavenging activity, β-carotene bleaching, inhibition of lipid peroxidation, reducing power ability, and cupric ion reducing antioxidant capacity (CUPRAC). Antioxidant potential of each mushroom species was calculated based on the average percentages relative to quercetin and summarized as Antioxidant Index (AI). Ganoderma lucidum (30.1%), Schizophyllum commune (27.6%), and Hericium erinaceus (17.7%) showed relatively high AI. Total phenolics in these mushrooms varied between 6.19 to 63.51 mg GAE/g extract. In the ACE inhibitory assay, G. lucidum was shown to be the most potent species (IC(50) = 50 μg/mL). Based on our findings, culinary-medicinal mushrooms can be considered as potential source of dietary antioxidant and ACE inhibitory agents.
  8. Fulltext The potential of mycelium and culture broth of Lignosus rhinocerotis as substitutes for the naturally occurring sclerotium with regard to antioxidant capacity, cytotoxic effect, and low-molecular-weight chemical constituents
    Lau BF, Abdullah N, Aminudin N, Lee HB, Yap KC, Sabaratnam V
    PLoS One, 2014;9(7):e102509.
    PMID: 25054862 DOI: 10.1371/journal.pone.0102509
    Previous studies on the nutritional and nutraceutical properties of Lignosus rhinocerotis focused mainly on the sclerotium; however, the supply of wild sclerotium is limited. In this investigation, the antioxidant capacity and cytotoxic effect of L. rhinocerotis cultured under different conditions of liquid fermentation (shaken and static) were compared to the sclerotium produced by solid-substrate fermentation. Aqueous methanol extracts of the mycelium (LR-MH, LR-MT) and culture broth (LR-BH, LR-BT) demonstrated either higher or comparable antioxidant capacities to the sclerotium extract (LR-SC) based on their radical scavenging abilities, reducing properties, metal chelating activities, and inhibitory effects on lipid peroxidation. All extracts exerted low cytotoxicity (IC50>200 µg/ml, 72 h) against selected mammalian cell lines. Several low-molecular-weight compounds, including sugars, fatty acids, methyl esters, sterols, amides, amino acids, phenolics, and triterpenoids, were identified using GC-MS and UHPLC-ESI-MS/MS. The presence of proteins (<40 kDa) in the extracts was confirmed by SDS-PAGE and SELDI-TOF-MS. Principal component analysis revealed that the chemical profiles of the mycelial extracts under shaken and static conditions were distinct from those of the sclerotium. Results from bioactivity evaluation and chemical profiling showed that L. rhinocerotis from liquid fermentation merits consideration as an alternative source of functional ingredients and potential substitute for the sclerotium.
  9. Sustainable landfill leachate treatment: Optimize use of guar gum as natural coagulant and floc characterization
    Cheng SY, Show PL, Juan JC, Ling TC, Lau BF, Lai SH, et al.
    Environ Res, 2020 09;188:109737.
    PMID: 32554270 DOI: 10.1016/j.envres.2020.109737
    Sustainable wastewater treatment necessitates the application of natural and green material in the approach. Thus, selecting a natural coagulant in leachate treatment is a crucial step in landfill operation to prevent secondary environmental pollution due to residual inorganic coagulant in treated effluent. Current study investigated the application of guar gum in landfill leachate treatment. Central composite design in response surface methodology was used to optimize the performance of Chemical Oxygen Demand (COD) removal. Quadratic model developed indicated the optimum COD removal 22.57% at guar gum dosage of 44.39 mg/L, pH 8.56 (natural pH of leachate) and mixing speed 79.27 rpm. Scanning electron microscopy showed that floc was compact and energy-dispersive-x-ray analysis showed that guar gum was capable to adsorb multiple ions from the leachate. Structural characterization using Fourier Transform Infrared analysis demonstrated that hydrogen bonding between guar and pollutant particles was involved in coagulation and flocculation process. Therefore, guar gum coagulant present potential to be an alternative in leachate treatment where pH requirement is not required during treatment. Simultaneously, adsorption by guar gum offers added pollutant removal advantage.
  10. New Prospects for Modified Algae in Heavy Metal Adsorption
    Cheng SY, Show PL, Lau BF, Chang JS, Ling TC
    Trends Biotechnol, 2019 Nov;37(11):1255-1268.
    PMID: 31174882 DOI: 10.1016/j.tibtech.2019.04.007
    Heavy metal pollution is one of the most pervasive environmental problems globally. Novel finely tuned algae have been proposed as a means to improve the efficacy and selectivity of heavy metal biosorption. This article reviews current research on selective algal heavy metal adsorption and critically discusses the performance of novel biosorbents. We emphasize emerging state-of-the-art techniques that customize algae for enhanced performance and selectivity, particularly molecular and chemical extraction techniques as well as nanoparticle (NP) synthesis approaches. The mechanisms and processes for developing novel algal biosorbents are also presented. Finally, we discuss the applications, challenges, and future prospects for modified algae in heavy metal biosorption.
  11. Anti-angiogenic and anti-tumour activities of Lignosus rhinocerus (Cooke) Ryvarden water extracts on HCT116 human colorectal carcinoma cells implanted in chick embryos
    Yong GY, Muniandy N, Beishenaliev A, Lau BF, Kue CS
    J Ethnopharmacol, 2024 Apr 16;331:118213.
    PMID: 38636576 DOI: 10.1016/j.jep.2024.118213
    ETHNOPHARMACOLOGICAL RELEVANCE: The sclerotium of Lignosus rhinocerus (Cooke) Ryvarden is used by the local communities in Southeast Asia and China to treat cancer, asthma, fever, and other ailments based on traditional knowledge. The sclerotial water extracts were previously reported to exhibit cytotoxic, apoptotic, and immunomodulatory activities - providing a scientific basis for its use in treating cancer; however, there is still a lack of evidence on its potential anti-angiogenic activity.

    AIM OF THE STUDY: This study aimed to investigate the toxicity, anti-angiogenic, and anti-tumour activities of the hot-water and cold-water extracts of L. rhinocerus using HCT116 human colorectal carcinoma cells implanted in the chick chorioallantoic membrane (CAM) model.

    MATERIALS AND METHODS: The toxicity of L. rhinocerus extracts towards the chick embryos was determined 24 h post-treatment. The anti-angiogenic activity of the extracts was then investigated at 0.1-10 μg/embryo (6.7-670 μg/mL) at targeted blood vessels. The anti-tumour effect of selected extracts against the HCT116 human colorectal carcinoma cells xenografted onto the chick embryos was also studied.

    RESULTS: The cold-water extracts of L. rhinocerus displayed strong in ovo toxicity (LC50: 1.2-37.7 μg/mL) while the hot-water extracts are non-toxic up to 670 μg/mL. Among the extracts, the hot-water extracts demonstrated the highest anti-angiogenic activity with 44.0 ± 17.7% reduction of capillary diameter (relative to the saline-treated control). Moreover, treatment of the HCT116 cells xenografted onto the chick embryos with the hot-water extracts resulted in smaller tumour size and lower number of blood vessels compared to the saline-treated control.

    CONCLUSIONS: The hot-water extracts of L. rhinocerus sclerotium demonstrated anti-angiogenic and anti-tumour activities but most of the cold-water extracts at similar concentrations were devoid of that. Our findings provide further scientific validation of the medicinal use of the sclerotium in treating cancer and thus, expanding our knowledge on the possible mechanism of its anti-cancer effect apart from direct cytotoxicity, induction of apoptosis and immunomodulation that have been studied thus far.

  12. Landfill leachate wastewater treatment to facilitate resource recovery by a coagulation-flocculation process via hydrogen bond
    Cheng SY, Show PL, Juan JC, Chang JS, Lau BF, Lai SH, et al.
    Chemosphere, 2021 Jan;262:127829.
    PMID: 32768754 DOI: 10.1016/j.chemosphere.2020.127829
    Recent trend to recover value-added products from wastewater calls for more effective pre-treatment technology. Conventional landfill leachate treatment is often complex and thus causes negative environmental impacts and financial burden. In order to facilitate downstream processing of leachate wastewater for production of energy or value-added products, it is pertinent to maximize leachate treatment performance by using simple yet effective technology that removes pollutants with minimum chemical added into the wastewater that could potentially affect downstream processing. Hence, the optimization of coagulation-flocculation leachate treatment using multivariate approach is crucial. Central composite design was applied to optimize operating parameters viz. Alum dosage, pH and mixing speed. Quadratic model indicated that the optimum COD removal of 54% is achieved with low alum dosage, pH and mixing speed of 750 mgL-1, 8.5 and 100 rpm, respectively. Optimization result showed that natural pH of the mature landfill leachate sample is optimum for alum coagulation process. Hence, the cost of pH adjustment could be reduced for industrial application by adopting optimized parameters. The inherent mechanism of pollutant removal was elucidated by FTIR peaks at 3853 cm-1 which indicated that hydrogen bonds play a major role in leachate removal by forming well aggregated flocs. This is concordance with SEM image that the floc was well aggregated with the porous linkages and amorphous surface structure. The optimization of leachate treatment has been achieved by minimizing the usage of alum under optimized condition.
  13. Recent developments on algal biochar production and characterization
    Yu KL, Lau BF, Show PL, Ong HC, Ling TC, Chen WH, et al.
    Bioresour Technol, 2017 Dec;246:2-11.
    PMID: 28844690 DOI: 10.1016/j.biortech.2017.08.009
    Algal biomass is known as a promising sustainable feedstock for the production of biofuels and other valuable products. However, since last decade, massive amount of interests have turned to converting algal biomass into biochar. Due to their high nutrient content and ion-exchange capacity, algal biochars can be used as soil amendment for agriculture purposes or adsorbents in wastewater treatment for the removal of organic or inorganic pollutants. This review describes the conventional (e.g., slow and microwave-assisted pyrolysis) and newly developed (e.g., hydrothermal carbonization and torrefaction) methods used for the synthesis of algae-based biochars. The characterization of algal biochar and a comparison between algal biochar with biochar produced from other feedstocks are also presented. This review aims to provide updated information on the development of algal biochar in terms of the production methods and the characterization of its physical and chemical properties to justify and to expand their potential applications.
  14. Zerumbone targets the CXCR4-RhoA and PI3K-mTOR signaling axis to reduce motility and proliferation of oral cancer cells
    Zainal NS, Gan CP, Lau BF, Yee PS, Tiong KH, Abdul Rahman ZA, et al.
    Phytomedicine, 2018 Jan 15;39:33-41.
    PMID: 29433681 DOI: 10.1016/j.phymed.2017.12.011
    BACKGROUND: The CXCR4-RhoA and PI3K-mTOR signaling pathways play crucial roles in the dissemination and tumorigenesis of oral squamous cell carcinoma (OSCC). Activation of these pathways have made them promising molecular targets in the treatment of OSCC. Zerumbone, a bioactive monocyclic sesquiterpene isolated from the rhizomes of tropical ginger, Zingiber zerumbet (L.) Roscoe ex Sm. has displayed promising anticancer properties with the ability to modulate multiple molecular targets involved in carcinogenesis. While the anticancer activities of zerumbone have been well explored across different types of cancer, the molecular mechanism of action of zerumbone in OSCC remains largely unknown.

    PURPOSE: Here, we investigated whether OSCC cells were sensitive towards zerumbone treatment and further determined the molecular pathways involved in the mechanism of action.

    METHODS: Cytotoxicity, anti-proliferative, anti-migratory and anti-invasive effects of zerumbone were tested on a panel of OSCC cell lines. The mechanism of action of zerumbone was investigated by analysing the effects on the CXCR4-RhoA and PI3K-mTOR pathways by western blotting.

    RESULTS: Our panel of OSCC cells was broadly sensitive towards zerumbone with IC50 values of less than 5 µM whereas normal keratinocyte cells were less responsive with IC50 values of more than 25 µM. Representative OSCC cells revealed that zerumbone inhibited OSCC proliferation and induced cell cycle arrest and apoptosis. In addition, zerumbone treatment inhibited migration and invasion of OSCC cells, with concurrent suppression of endogenous CXCR4 protein expression in a time and dose-dependent manner. RhoA-pull down assay showed reduction in the expression of RhoA-GTP, suggesting the inactivation of RhoA by zerumbone. In association with this, zerumbone also inhibited the PI3K-mTOR pathway through the inactivation of Akt and S6 proteins.

    CONCLUSION: We provide evidence that zerumbone could inhibit the activation of CXCR4-RhoA and PI3K-mTOR signaling pathways leading to the reduced cell viability of OSCC cells. Our results suggest that zerumbone is a promising phytoagent for development of new therapeutics for OSCC treatment.

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