Differential Analysis of Korean and Chinese Lingzhi or Reishi Medicinal Mushroom Ganoderma lucidum (Agaricomycetes) Spore Powder by Infrared Spectroscopy with Stoichiometry.

To investigate the differences between Korean Ganoderma lucidum spore powder (KP), broken-spo-roderm KP (BSKP), Chinese traditional G. lucidum spore powder (CP), and broken-sporoderm CP (BSCP), they were identified by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), second derivative infrared spectroscopy (SD-IR), dual-index sequence analysis (DISA) and X-ray diffraction (XRD). SEM showed that there were no significant differences in microstructure between the two kinds of spore powders. FT-IR spectra showed that the four spore powders appeared with characteristic peaks of 3400, 3006, 2925, 1745, 1535, 1454, 1249, 1074, 1049, and 896 cm-1, respectively, they were contained the characteristic peaks of total triterpenes, polysaccharides and fatty acids. DISA showed that the same species of spore powders, the overall similarity of before and broken the sporoderm was high with minor differences and there were no differences between the different kinds of spore powders. Similarity analysis showed that the four spore powders were in high agreement and were no differences. The polysaccharide, total triterpene, spore oil and protein content of the four spore powders were determined separately. The results showed that the active ingredients content of the batch of KP were lower than that of CP, that of BSKP were lower than that of BSCP, while the active ingredients content of both broken-sporoderm spore powders were higher than that of before broken-sporoderm. It is inferred that the structure of the main chemical and component of KP is the same as that of CP. This study provides a reference for the future development and application of G. lucidum.

Water-based Fe3O4 magnetic fluid-coated Aspergillus niger spores for treating liquid contaminated with Cr(VI).

The use of magnetic biosorbents for the remediation of heavy metals has attracted increasing attention due to their ease of separation and reusability. We developed a method for preparing superparamagnetic biosorbent materials using water-based magnetic fluids. Water-based magnetic fluid-spores (WMFSs) were obtained by combining water-based magnetic fluid (WMF) with Aspergillus niger spores at ratios of 0.6:1 (WMFS1), 0.8:1 (WMFS2), 1:1 (WMFS3), 1.2:1 (WMFS4), and 1.4:1 (WMFS5). A magnetic composite material was prepared from magnetic nanoparticles and spores in a ratio of 1:1 as a control. The adsorption efficiency and separation effect of WMFS3 were significantly better than those of the magnetic composite material. The morphology and structure of WMFS3 were characterized by performing transmission electron microscopy. The results showed that Fe3O4 magnetic particles were uniformly coated on the spore surface. The superparamagnetism of WMFS3 was tested using a vibrating sample magnetometer. At pH 2.0, the maximum adsorption capacity of WMFS3 for Cr(VI) was 105 mg/g; in the pH range of 2.0-3.0, the adsorption equilibrium time of WMFS3 was 60 min. Thus, the adsorption process conformed to the pseudo-second-order kinetic model and Freundlich isotherm. Thermodynamic studies showed that the process was spontaneous and endothermic. The adsorption mechanisms of WMF3 for Cr(VI) included electrostatic, reduction, and complexation adsorption. This biosorbent material showed excellent adsorption performance for Cr(VI) and is promising for wastewater resource applications.

Association of airborne particulate matter with pollen, fungal spores, and allergic symptoms in an arid urbanized area.

Studies focused on the seasonal distribution of pollen and spores in semiarid cities are scarce. At these sites, climate change potentiates the emission and transport of fine (PM10) to ultrafine particles (PM2.5), easily attached to pollen surfaces, causing allergen's release. This study examines the potential correlation of seasonal variations of pollen, fungal spores, PM10, and meteorological parameters with allergic reactions of 150 people living in a Sonoran desert city. We collected PM10, airborne pollen, and spores during a year. We also studied topsoil and road dust samples as potential PM-emission sources. We obtained dust-mineralogy, chemistry, and particle size attached to pollen by X-ray diffraction and scanning electron microscope. Results show that seasonal high PM-loading in the urban atmosphere coincides with aeroallergens promoting micro- to nanoparticles' attachment to pollen's surface. A collapsed membrane was observed in several samples after individual grains show the following maximum wall coverage: Poaceae 28%, Asteraceae 40%, Chenopodiaceae-Amaranthacea 29%, Fabaceae 18%. Most of the particles covering pollen's surface have a geogenic origin mixed with metals linked to traffic (bromide, chlorine, and antimony). Mineralogical, granulometric analysis, and main wind-direction show that two local soil-types are the main contributors to PM. A high frequency of positive sensitization to pollen with high particle loading was detected. These results suggest that climate-driven dust emissions may alter pollen and spore surfaces' physicochemical characteristics with the further consequences in their allergenic potential.

Prophylactic effects of sporoderm-removed Ganoderma lucidum spores in a rat model of streptozotocin-induced sporadic Alzheimer's disease.

ETHNOPHARMACOLOGICAL RELEVANCE: Ganoderma lucidum (G. lucidum, Lingzhi), also known as "immortality mushroom" has been broadly used to improve health and longevity for thousands of years in Asia. G. lucidum and its spores have been used to promote health, based on its broad pharmacological and therapeutic activity. This species is recorded in Chinese traditional formula as a nootropic and has been suggested to improve cognitive dysfunction in Alzheimer's disease. However, little is known about the nootropic effects and molecular mechanism of action of G. lucidum spores. AIM OF THE STUDY: The present study investigated the protective effects of sporoderm-deficient Ganoderma lucidum spores (RGLS) against learning and memory impairments and its mechanism of action. MATERIALS AND METHODS: In the Morris water maze, the effects of RGLS on learning and memory impairments were evaluated in a rat model of sporadic Alzheimer's disease that was induced by an intracerebroventricular injection of streptozotocin (STZ). Changes in amyloid ß (Aß) expression, Tau expression and phosphorylation, brain-derived neurotrophic factor (BDNF), and the BDNF receptor tropomyosin-related kinase B (TrkB) in the hippocampus were evaluated by Western blot. RESULTS: Treatment with RGLS (360 and 720 mg/kg) significantly enhanced memory in the rat model of STZ-induced sporadic Alzheimer's disease and reversed the STZ-induced increases in Aß expression and Tau protein expression and phosphorylation at Ser199, Ser202, and Ser396. The STZ-induced decreases in neurotrophic factors, including BDNF, TrkB and TrkB phosphorylation at Tyr816, were reversed by treatment with RGLS. CONCLUSION: These findings indicate that RGLS prevented learning and memory impairments in the present rat model of STZ-induced sporadic Alzheimer's disease, and these effects depended on a decrease in Aß expression and Tau hyperphosphorylation and the modulation of BDNF-TrkB signaling in the hippocampus.

Polysaccharide from spore of Ganoderma lucidum ameliorates paclitaxel-induced intestinal barrier injury: Apoptosis inhibition by reversing microtubule polymerization.

Side effects of chemotherapy are burning questions for physicians and patients involved in cancers. Ganoderma lucidum is a widely consumed traditional Chinese medicine and edible mushroom with multiple functional properties. The present study aims to investigate the potential of polysaccharides from spore of G. lucidum (SGP) on small intestinal barrier function recovery against paclitaxel (PTX) challenge in a breast cancer mice model and IEC-6 cell line. The 4T1 tumor-bearing mice were treated with PTX together with four-week daily oral administration of SGP. Results indicated that combination of PTX and SGP reversed body weight lost and remolded the histology of small intestine, accompanied with promoted proliferation but suppressed apoptosis in intestinal cells. Intestinal barrier function was enhanced by the combination as indicated by reduced endotoxemia and the up-regulation of tight junction proteins, including Zonula occludens-1 (ZO-1), E-cadherin, ß-catenin and Occludin. The protection of SGP was further confirmed in IEC-6 cells affected by PTX in vitro. The combination treatment prevented PTX-induced apoptosis in IEC-6 by inhibiting microtubule polymerization, and the aforementioned tight junction proteins were also upregulated. These findings suggest a promising protective effect of SGP against small intestinal barrier injury caused by PTX, highlighting its clinical implication against the chemotherapy side effects.

Structural Analysis and Antioxidant Activity of Extracellular Polysaccharides Extracted from Culinary-Medicinal White Jelly Mushroom Tremella fuciformis (Tremellomycetes) Conidium Cells.

Using Tremella fuciformis conidium cells for submerged fermentation is a cost-effective way to harvest bioactive compounds. In this study, we emphasized the structural and functional analysis of extracellular polysaccharides (EPS) extracted from T. fuciformis conidium cells. An EPS high-yield strain tyc63 was selected and a 6-day optimum fermentation period was determined. Crude EPS was extracted and three high molecular weight (5189, 171.6, and 661 kDa) polysaccharides TFP-1, TFP-2, and TFP-3 were isolated and purified. TFP-1 is mainly composed of glucose, xylose, mannose, and fucose, while both TFP-2 and TFP-3 are mainly composed of rhamnose, arabinose, mannose, galactose, and glucose. FT-IR analysis revealed that TFP-1, TFP-2, and TFP-3 have typical polysaccharide structure. The antioxidant assay revealed that the crude EPS, TFP-1, TFP-2, and TFP-3 presented high free radical scavenging activities but low ferric reducing power, suggesting that the EPS produced by liquid fermentation could be used as a potent radical scavenger.

The effect of Ganodermalucidum spore oil in early skin wound healing: interactions of skin microbiota and inflammation.

The mushroom Ganoderma lucidum (G. lucidum Leyss. ex Fr.) Karst has been a traditional Chinese medicine for millennia. In this study, we isolated the Ganoderma lucidum spore oil (GLSO) and evaluated the effect of GLSO on skin burn wound healing and the underlying mechanisms. Mice were used to perform skin wound healing assay. Wound analysis was performed by photography, hematoxylin/eosin staining, Masson's Trichrome staining and immunohistochemical analysis. Microbiota on the wounds were analyzed using the 16s rRNA sequence and quantitative statistics. The lipopolysaccharide (LPS) content was examined in skin wounds and serum using an enzyme-linked immunosorbent assay (ELISA). The expression of Toll-like receptor 4 (TLR4) and the relative levels of inflammatory cytokines were determined by qPCR and immunofluorescence assay. A pseudo-germfree mouse model treated with antibiotics was used to investigate whether GLSO accelerated skin burn wound healing through the skin microbiota. We found that GLSO significantly accelerated the process of skin wound healing and regulated the levels of gram-negative and gram-positive bacteria. Furthermore, GLSO reduced LPS and TLR4, and levels of some other related inflammatory cytokines. The assay with the pseudo-germfree mice model showed that GLSO had a significant acceleration on skin wound healing in comparison with antibiotic treatment. Thus, GLSO downregulated the inflammation by regulating skin microbiota to accelerate skin wound healing. These findings provide a scientific rationale for the potential therapeutic use of GLSO in skin burn injury.

Solid-state fermentation for enhanced production of selenium nanoparticles by gamma-irradiated Monascus purpureus and their biological evaluation and photocatalytic activities.

Selenium nanoparticles (SeNPs) were successfully synthesized using the culture extract of Monascus purpureus ATCC16436 grown on sugarcane bagasse under solid-state fermentation. The rapid synthesis of SeNPs was completed after 30 min as confirmed by UV-Vis spectroscopy. Functional groups present in the synthesized SeNPs samples were confirmed by Fourier transform infrared spectroscopy. The synthesized SeNPs showed a single-phase crystalline structure. Transmission electron microscope revealed the spherical shape and the mean particle size was 46.58 nm. Dynamic light scattering analysis showed that the synthesized SeNPs were monodispersed and the recorded polydispersity index value was 0.205. Zeta potential value of - 24.01 mV indicated the high stability of SeNPs. Besides, the biological activities of antioxidant, anticancer and antimicrobial as well as the photocatalytic activities were also studied. SeNPs showed promising antioxidant activity with 50% inhibitory concentration of 85.92 µg mL-1. Based on the MTT assay, SeNPs inhibited the proliferation of normal human melanocytes, human breast and liver cancer cell lines with 50% inhibitory concentrations of 45.21, 61.86 and 200.15 µg mL-1, respectively. SeNPs showed broad spectrum of antimicrobial potential against the tested human and plant pathogens. SeNPs showed efficient degradation of methylene blue dye. Moreover, the effect of gamma irradiation on the production enhancement of SeNPs was also adopted. Exposure of the fungal spores to gamma rays at 1000 Gy increased the yield of SeNPs to approximately fivefold. Hence, this study suggests a new and alternate approach with the excellent biotechnological potentiality for the production of SeNPs.

Ganoderma lucidum spore oil induces apoptosis of breast cancer cells in vitro and in vivo by activating caspase-3 and caspase-9.

ETHNOPHARMACOLOGICAL RELEVANCE: The mushroom Ganoderma lucidum (G. lucidum) is a traditional Chinese medicine reported to have a variety of pharmacological properties, including anti-cancer activity. G. lucidum spore oil (GLSO) is a lipid substance extracted from sporoderm-broken spore of G. lucidum. However, the effect of GLSO on breast cancer and the underlying molecular mechanism remain unclear. AIM OF THE STUDY: The aim of this study was to identify the effects of GLSO on breast cancer cells in vitro and in vivo as well as to investigate the mechanistic basis for the anticancer effect of GLSO. MATERIALS AND METHODS: First, in vitro MDA-MB-231 cells were treated with GLSO (0.2, 0.4, and 0.6 µL/mL). The protein levels of B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), X-linked inhibitor of apoptosis (XIAP), total poly (ADP-ribose) polymerase (PARP), caspase-3 and caspase-8 were examined using western blotting. The mRNA expression levels of Fas-associated protein with death domain (FADD), TNF receptor-associated factor 2 (TRAF2), caspases-3, -8, -9 and Bax were examined using qRT-PCR. Second, in vivo the anticancer properties of GLSO were assessed by H&E, TUNEL and immunohistochemistry in BALB/c mice injected with 4T1 cells. In addition, the levels of caspase-9/caspase-3 signaling pathway proteins in tumor tissue were evaluated by immunoblotting. Finally, MDA-MB-231 cells were treated with caspase inhibitors to measure cell viability, the protein levels were examined with western blotting. RESULTS: The results in vitro showed that GLSO up-regulated the expression of Bax and caspase-3 in MDA-MB-231 cells, but had no effect on the expression of caspase-8. Moreover, the growth of tumors in vivo was significantly suppressed in the GLSO-treated group. The results of Western blot were consistent with in vitro. In vitro, co-treatment of MDA-MB-231 cells with caspase inhibitors reduced the inhibitory effect of GLSO on cell growth. CONCLUSIONS: GLSO inhibits the growth of MDA-MB-231 cells and tumors in vivo by inducing apoptosis, which may be achieved through the mitochondrial apoptotic pathway.

Facile Nanolization Strategy for Therapeutic Ganoderma Lucidum Spore Oil to Achieve Enhanced Protection against Radiation-Induced Heart Disease.

Radiotherapy (RT) has been extensively utilized for clinical cancer therapy, however, excessive generation of reactive oxygen species (ROS) is becoming a main cause for radiation-induced heart disease (RIHD). Ganoderma lucidum spore oil (GLSO) is a popular functional food composite with potent antioxidant activity, but it is compromised by poor solubility and stability for further application. Therefore, a strategy for rational fabrication of GLSO@P188/PEG400 nanosystem (NS) is demonstrated in this study to realize good water solubility and achieve enhanced protection against RIHD. As expected, GLSO@P188/PEG400 NS can attenuate X-ray-induced excessive ROS levels thanks to its enhanced free radical scavenging capability, simultaneously protecting on mitochondria from X-ray irradiation (IR). Moreover, GLSO@P188/PEG400 NS alleviates DNA damage and promotes self-repair processes against IR, thus recovering G0/G1 proportion back to normal levels. Furthermore, pre- and post-treated GLSO@P188/PEG400 NS demonstrates potential protection on heart from X-rays in vivo, as evidenced by attenuating cardiac dysfunction and myocardial fibrosis. Meanwhile, the cell antioxidant capacity (including T-SOD, MDA, and GSH-x) stays in balance during this process. This study not only provides a promising strategy for facile nanolization of functional food composites with hydrophobic defects but also sheds light on their cardiac protection and action mechanisms against IR-induced disease.

Quantitative analysis of diverse sporomorph-derived sporopollenins.

Over the years studies on sporopollenin have reported a wide variety of structures. However, the methods and techniques used to elucidate sporopollenin structures are highly diverse so that much is still unclear with respect to the nature and structural diversity of sporopollenins. In order to investigate the structural diversity in sporopollenin between different taxa, extant sporomorphs of ten different species ranging from a mushroom to a cycad were examined using a relatively simple and fast analytical procedure. Sporomorphs, before and after saponification, were analysed for sporopollenin composition by Thermally assisted Hydrolysis and Methylation (THM) using [13C]tetramethylammonium hydroxide ([13C]TMAH). The sporomorp chemical composition differed markedly between the groups of organisms analysed. Moreover, we not only identified the nature and relative quantities of the well-known sporopollenin constituents p-coumaric acid and ferulic acid but also many other phenolic moieties, such as caffeic acid, which appeared to be the most abundant phenolic constituent in spores of Equisetum palustre, Salvinia molesta, Cyrtomium falcatum and Anemia phyllitidis. Within the two Equisetum species analysed as well as in the closely related Azolla and Salvinia species the same suite of phenolic constituents were observed, but their relative distribution varied largely. We thus demonstrate the existence of a high structural diversity, both qualitatively and quantitatively in sporopollenins enabling future studies related to the evolution, phylogeny and (palaeo)environment of sporopollenin-producing organisms. Furthermore, a better knowledge of sporopollenin and its structural variety is of relevance to the rapidly growing application of spores and pollen as a drug delivery agent in medicine.

Synthesis and Evaluation of Herbal Chitosan from Ganoderma Lucidum Spore Powder for Biomedical Applications.

Chitosan is an extremely valuable biopolymer and is usually obtained as a byproduct from the shells of crustaceans. In the current work, chitosan is obtained from an herbal source (Ganoderma lucidum spore powder (GLSP)) for the first time. To show this, both standard (thermochemical deacetylation, (TCD)) and emerging (ultrasound-assisted deacetylation (USAD)) methods of chitosan preparation were used. The obtained chitosan was characterized by elemental analysis, XRD (X-ray diffraction), FT-IR (Fourier transform infrared spectroscopy) and thermogravimetric measurements. The process resulted in chitosan possessing comparable values of DD, [η] and [Formula: see text] to the commercial product. Chitosan obtained via both processes (TCD and USAD) displayed excellent biocompatibility; although the USAD prepared biopolymer exhibited significantly improved fibroblast (L929 cell) viability and enhanced antibacterial zones for both Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). The findings of new herbal chitosan mark key developments of natural biomaterials; marking a potential shift from conventional sea-based organisms.

Morphological Characteristics of Monokaryotic and Dikaryotic Collections of Three Medicinal Coprinellus Species (Agaricomycetes).

This article presents data from morphological observations of mycelia of 40 monokaryotic and 11 dikaryotic collections of 3 medicinal Coprinellus species (C. disseminatus, C. micaceus, and C. xanthothrix). The growth rate, colony morphology, and micromorphological characteristics of mycelia and anamorphs on 1.5% malt-extract agar (MEA) and potato-dextrose agar (PDA) are described. Well-developed white, cottony-felt colonies, which later show creamy, yellowish to rusty brown pigmentation on mycelia and agar, were typical for the studied Coprinellus collections. Mycelial growth was denser on PDA than on MEA, whereas the average growth rate indicators (GRavr) were higher in dikaryotic isolates on MEA. Clamp connections were described only in dikaryotic isolates of C. disseminatus and C. micaceus; mycelia of C. xanthothrix had no clamps. Nonsporulating Ozonium-type anamorphic mycelia (with a rusty brown septate and parallel hyphal strands), a taxonomic feature characteristic of the clade Coprinellus, was present in the studied monokaryotic and dikaryotic collections, whereas Hormographiella-type sporulating anamorphs developed only in monokaryotic and dikaryotic isolates of C. xanthothrix. Yellowish-rusty-brownish regular hyphal loops were also observed in the collections of all 3 Coprinellus species. Allocyst-like hyphal swellings were observed in monokaryons of C. xanthothrix, and hyphocystidia were observed in dikaryons of C. micaceus. Hyphal loops and hyphal cystidia presumably were derived from Ozonium mycelia. Thick-walled, oval chlamydospores and chlamydospore-like swellings were described only in dikaryons of C. xanthothrix. Under these experimental conditions, primordia and fruiting bodies developed in dikaryons of C. xanthothrix on MEA and PDA, respectively, and in dikaryons of C. micaceus on MEA. The taxonomic significance of the mycelial and anamorphic characteristics of studied Coprinellus species was evaluated. They could be useful for identifying mycelial cultures during biotechnological cultivation.

Aeroallergens in North-Central Nigeria.

INTRODUCTION AND OBJECTIVES: Aeroallergens are airborne organic substances which are responsible for allergenic diseases in hypersensitive individuals. People are exposed to their allergens either directly or after their entrance into the interiors. The spatio-temporal pattern of aeroallergens and their relationship with weather variability in Abuja and Nassarawa, North-Central Nigeria was studied. MATERIALS AND METHODS: Aerosamples were trapped with modified Tauber-like pollen traps. Samples were collected monthly and centrifuged at 2500rpm for 5 min and subjected to acetolysis. Meteorological data were collected from the Nigerian Meteorological Agency. RESULTS AND CONCLUSION: Aeroallergens concentration were unequivocally regulated by weather variables in both locations, indicating the possible use of aeroallergens especially pollen and spores as bio-indicators of weather variations and change. Aeroallergens encountered were fungal spores, pollen, diatom frustules, fern spores, algal cyst/cells in decreasing order of dominance. Among pollen group, Poaceae, Amarathaceae/Chenopodiaceae and Hymenocardia acida dominated. Spores of Smut species, Puccinia, Curvularia and Nigrospora were major contributors among aeromycoflora. Fungal spores morphotype dominated during the rainier months and were major contributors of the aeroallergen spectrum with those belonging to Deuteromycete preponderant. Aeroallergens which were previously identified as triggers of conjunctivitis, asthma, allergic sinusitis and bronchopulmonary allergic diseases were frequently present in both locations. Pollen prevailed more during the harmattan, influenced by northeast trade wind. Pollen component differed and was based on autochthonous source plants, indicating difference in sub-vegetational types.

Naphthoquinone Metabolites Produced by Monacrosporium ambrosium, the Ectosymbiotic Fungus of Tea Shot-Hole Borer, Euwallacea fornicatus, in Stems of Tea, Camellia sinensis.

The tea shot-hole borer beetle (TSHB, Euwallacea fornicatus) causes serious damage in plantations of tea, Camellia sinensis var. assamica, in Sri Lanka and South India. TSHB is found in symbiotic association with the ambrosia fungus, Monacrosporium ambrosium (syn. Fusarium ambrosium), in galleries located within stems of tea bushes. M. ambrosium is known to be the sole food source of TSHB. Six naphthoquinones produced during spore germination in a laboratory culture broth of M. ambrosium were isolated and identified as dihydroanhydrojavanicin, anhydrojavanicin, javanicin, 5,8-dihydroxy-2-methyl-3-(2-oxopropyl)naphthalene-1,4-dione, anhydrofusarubin and solaniol. Chloroform extracts of tea stems with red-colored galleries occupied by TSHB contained UV active compounds similar to the above naphthoquinones. Laboratory assays demonstrated that the combined ethyl acetate extracts of the fungal culture broth and mycelium inhibited the growth of endophytic fungi Pestalotiopsis camelliae and Phoma multirostrata, which were also isolated from tea stems. Thus, pigmented naphthoquinones secreted by M. ambrosium during spore germination may prevent other fungi from invading TSHB galleries in tea stems. The antifungal nature of the naphthoquinone extract suggests that it protects the habitat of TSHB. We propose that the TSHB fungal ectosymbiont M. ambrosium provides not only the food and sterol skeleton necessary for the development of the beetle during its larval stages, but also serves as a producer of fungal inhibitors that help to preserve the purity of the fungal garden of TSHB.

Biological characteristics, bioactive components and antineoplastic properties of sporoderm-broken spores from wild Cordyceps cicadae.

BACKGROUND: Cordyceps cicadae, an entomogenous fungus has been used as a dietary therapeutic in traditional Chinese medicine for several millennia, in the form of powders and decoction. However, wild C. cicadae is notably scarce. To date, there is still a lack of comprehensive and deep studies on the biological characteristics, chemical profiles and antineoplastic mechanisms of C. cicadae, especially its spores. AIM OF THE STUDY: This study aimed to identify wild C. cicadae using rDNA-ITS sequences. Active constituents and volatile ingredients of C. cicadae sporoderm-broken spore powders (CCBSP) were elucidated using UPLC-ESI-Q-TOF-MS and GC-MS, respectively. The underlying anti-neoplastic mechanisms of CCBSP were further investigated in A549 lung carcinoma cells. RESULTS: Molecular phylogenetic analysis of nuclear rDNA sequences indicated that wild C. cicadae belonged to Paecilomyces cicadae. Eight primary compounds from CCBSP were identified by MS fragmentation ions including nucleosides, cordycepic acid, cordycepin, beauvericin and myriocin. In total, forty-nine volatile components representing 99.56% of CCBSP were clearly identified. CCBSP exhibited antiproliferative effects on A549 cells with IC50 value of 125.54 ± 2.71 µg/ml, blocking the cell cycle in the G2/M phase. The nuclear morphology exhibited typical characteristics of apoptosis by Hoechst fluorescent stain. AnnexinV-FITC/PI staining revealed that the number of apoptotic cells increased after CCBSP treatment. Furthermore, immunofluorescence experiments indicated that CCBSP lowered the expressions of ß-catenin and N-cadherin, which was accompanied by repressed Wnt/ß-catenin signalling and activation of caspase-mediated apoptosis pathways. CONCLUSIONS: rDNA-ITS sequencing enabled molecular identification of wild C. cicadae. Importantly, these findings provide the first evidence regarding the full-scale bioactive components and antineoplastic properties of CCBSP. These data highlight the significance of C. cicadae as a potential antineoplastic agent.

Image Cytometric Analysis of Algal Spores for Evaluation of Antifouling Activities of Biocidal Agents.

Chemical biocides have been widely used as marine antifouling agents, but their environmental toxicity impose regulatory restriction on their use. Although various surrogate antifouling biocides have been introduced, their comparative effectiveness has not been well investigated partly due to the difficulty of quantitative evaluation of their antifouling activity. Here we report an image cytometric method to quantitatively analyze the antifouling activities of seven commercial biocides using Ulva prolifera as a target organism, which is known to be a dominant marine species causing soft fouling. The number of spores settled on a substrate is determined through image analysis using the intrinsic fluorescence of chlorophylls in the spores. Pre-determined sets of size and shape of spores allow for the precise determination of the number of settled spores. The effects of biocide concentration and combination of different biocides on the spore settlement are examined. No significant morphological changes of Ulva spores are observed, but the amount of adhesive pad materials is appreciably decreased in the presence of biocides. It is revealed that the growth rate of Ulva is not directly correlated with the antifouling activities against the settlement of Ulva spores. This work suggests that image cytometric analysis is a very convenient, fast-processable method to directly analyze the antifouling effects of biocides and coating materials.

Demulsification of crude oil-in-water emulsions by means of fungal spores.

The present feature describes for the first time the application of spores from Aspergillus sp. IMPMS7 to break out crude oil-in-water emulsions (O/W). The fungal spores were isolated from marine sediments polluted with petroleum hydrocarbons. The spores exhibited the ability to destabilize different O/W emulsions prepared with medium, heavy or extra-heavy Mexican crude oils with specific gravities between 10.1 and 21.2°API. The isolated fungal spores showed a high hydrophobic power of 89.3 ± 1.9% and with 2 g of spores per liter of emulsion, the half-life for emulsion destabilization was roughly 3.5 and 0.7 h for extra-heavy and medium crude oil, respectively. Then, the kinetics of water separation and the breaking of the O/W emulsion prepared with heavy oil through a spectrofluorometric technique were studied. A decrease in the fluorescence ratio at 339 and 326 nm (I339/I326) was observed in emulsions treated with spores, which is similar to previously reported results using chemical demulsifiers.

Pharmacological Properties of Biocompounds from Spores of the Lingzhi or Reishi Medicinal Mushroom Ganoderma lucidum (Agaricomycetes): A Review.

Ganoderma lucidum is a well-known representative of mushrooms that have been used in traditional Chinese medicine for centuries. New discoveries related to this medicinal mushroom and its biological properties are frequently reported. However, only recently have scientists started to pay special attention to G. lucidum spores. This is in part because of the recent development of methods for breaking the spore wall and extracting biocompounds from the spore. Although some research groups are working with G. lucidum spores, data in the literature are still limited, and the methods used have not been systematized. This review therefore describes the main advances in techniques for breaking the spore wall and extracting biocompounds from the spore. In addition, the major active components identified and their biological properties, such as neurological activity and antiaging and cell-protective effects, are investigated because these are of importance for potential drug development.

Sporoderm-Broken Spores of Ganoderma lucidum Inhibit the Growth of Lung Cancer: Involvement of the Akt/mTOR Signaling Pathway.

The sporoderm-broken spores of Ganoderma lucidum (SBGS) and their extracts exhibited a wide range of biological activities. In the present study, we prepare ethanol/ethanol extract (E/E-SBGS) and ethanol/aqueous extract (E/A-SBGS) from SBGS and examine their antitumor activities against human lung cancer. Our results showed that E/E-SBGS, not E/A-SBGS, inhibited the survival and migration of lung cancer cells in a dose-dependent manner. E/E-SBGS arrested cell cycle at G2/M phase and triggered apoptosis by decreasing the expression and activity of cell cycle regulators, cyclin B1 and cdc2, as well as anti-apoptotic proteins, Bcl-2 and Bcl-xl. Consequently, colony formation of lung cancer cells was markedly blocked by E/E-SBGS at subtoxic concentrations. Oral administration of both E/E-SBGS and SBGS significantly suppressed tumor volume and tumor weight without gross toxicity in mice. Mechanism study showed that E/E-SBGS dose-dependently suppressed the activation of Akt, the mammalian target of rapamycin (mTOR) and their downstream molecules S6 kinase and 4E-BP1 in treated tumor cells. Taken together, these results indicate that the ethanol extract of sporoderm-broken spores of G. lucidum suppresses the growth of human lung cancer, at least in part, through inhibition of the Akt/mTOR signaling pathway, suggesting its potential role in cancer treatments.