Evaluation of the Hepatoprotective Effect of the Lumpy Bracket Medicinal Mushroom Trametes gibbosa (Agaricomycetes) on CCl4-Induced Liver Injury in Rats.

Mushrooms have been used as medicine by humans for more than 5000 years. They have had a successful role in treating immune deficiencies. Nowadays, some extracts and compounds obtained from medicinal mushrooms have increased a great prospect of treating many disorders by having a great role in modulation of immune system, cancer inhibiting, cardio-vascular health, antiviral, antibacterial, antioxidant and protective effects against hepatitis and diabetes. In this study, we evaluated the antioxidant effect of methanol and hot water extract of the Trametes gibbosa (Pers.) Fr. mushroom and hepatoprotective effect of the extract with the most radical scavenging potency. To assess the antioxidant properties of different extracts of the mushroom, DPPH method was used. For assessing the hepatoprotective properties, a seven-day experiment was designed, and liver toxicity was induced by carbon tetrachloride [intraperitoneal (ip) for 7 consecutive days, 0.5 mL/kg body weight (BW)]. Rats were simultaneously fed with aqueous extract of the mushroom with the dose of 250, 500, and 1000 mg/kg BW and silymarin (100 mg/kg BW) as positive control. At the end of the experiment, blood serums of the rats were collected for quantification of major liver factors (e.g., aspartate aminotransferase, alanine aminotransferase, alanine phosphatase, bilirubin, etc.). Tissue samples were obtained for pathological examination. Based on the results, the aqueous extract showed more potent radical scavenging activity (half-maximal inhibitory concentration = 414.33 µg/mL, compared with 936.92 µg/mL for methanolic extract). Indeed, hepatoprotective properties of the aqueous extract of the mushroom (500 and 1000 mg/kg BW) were comparable with those of silymarin and even showed superior protective effects in histopathological examination. It seems that with further complementary studies, T. gibbosa could be considered a potential candidate for hepatoprotection.

Huaier enhances the tumor-killing effect and reverses gemcitabine-induced stemness by suppressing FoxM1.

BACKGROUND: Gemcitabine is the first-line chemotherapy drug that can easily cause chemotherapy resistance. Huaier is a traditional Chinese medicine and shows an antitumor effect in pancreatic cancer, but whether it can enhance the gemcitabine chemotherapeutic response and the potential mechanism remain unknown. PURPOSE: This study was performed to explore the effect of Huaier in promoting the tumor-killing effect of gemcitabine and elucidate the possible mechanism in pancreatic cancer. METHODS: Cell Counting Kit-8 assays and colony formation assays were used to detect proliferation after different treatments. Protein coimmunoprecipitation was applied to demonstrate protein interactions. Nuclear protein extraction and immunofluorescence were used to confirm the intracellular localization of the proteins. Western blotting was performed to detect cell proliferation-related protein expression or cancer stem cell-associated protein expression. Sphere formation assays and flow cytometry were used to assess the stemness of pancreatic cancer cells. The in vivo xenograft model was used to confirm the inhibitory effect under physiological conditions, and immunohistochemistry was used to detect protein expression. RESULTS: Huaier suppressed the proliferation and stem cell-like properties of pancreatic cancer cells. We found that Huaier suppressed the expression of forkhead box protein M1 (FoxM1). In addition, Huaier inhibited FoxM1 function by blocking its nuclear translocation. Treatment with Huaier reversed the stemness induced by gemcitabine in a FoxM1-dependent manner. Furthermore, we verified the above results by an in vivo study, which reached the same conclusion as those in vitro. CONCLUSION: Overall, this study illustrates that Huaier augments the tumor-killing effect of gemcitabine through suppressing the stemness induced by gemcitabine in a FoxM1-dependent way. These results indicate that Huaier can be applied to overcome gemcitabine resistance.

Using low-shear aerated and agitated bioreactor for producing two specific laccases by trametes versicolor cultures induced by 2,5-xylidine: Process development and economic analysis.

Laccase isoforms from basidiomycetes exhibit a superior redox potential compared to commercially available laccases obtained from ascomycete fungi, rendering them more reactive toward mono-substituted phenols and polyphenolic compounds. However, basidiomycetes present limitations for large-scale culture in liquid media, restraining the current availability of laccases from this fungal class. To advance laccase production from basidiomycetes, a newly designed 14-L low-shear aerated and agitated bioreactor provided enzyme titers up to 23.5 IU/mL from Trametes versicolor cultures. Produced enzymes underwent ultrafiltration and LC/MS-MS characterization, revealing the predominant production of only two out of the ten laccases predicted in the T. versicolor genome. Process simulation and economic analysis using SuperPro designer® suggested that T. versicolor laccase could be produced at US$ 3.60/kIU in a 200-L/batch enterprise with attractive economic parameters and a payback period of 1.7 years. The study indicates that new bioreactors with plain design help to produce low-cost enzymes from basidiomycetes.

Structure elucidation and antiviral activity of a cold water-extracted mannogalactofucan Ts1-1A from Trametes sanguinea against human cytomegalovirus in vitro.

In this study, we purified a partially acetylated heteropolysaccharide (Ts1-1A) from the fruit bodies of Trametes sanguinea Lloyd through cold water extraction and serial chromatographic separation. The purified polysaccharide Ts1-1A (12.8 kDa) was characterized as a branched mannogalactofucan with a backbone of alternately connected 1,3-linked α-Fucp and 1,6-linked α-Galp, which was partially substituted by non-reducing end units of ß-Manp at O-2 and O-3 positions of 1,6-linked α-Galp. Ts1-1A showed pronounced anti-human cytomegalovirus activity at the concentration of 200 and 500 µg/mL in systematical assessments including morphological changes, western blotting, qPCR, indirect immunofluorescence and tissue culture infective dose assays. Moreover, Ts1-1A exerted its antiviral activity at two distinct stages of viral proliferation manifesting as significantly inhibiting viral protein (IE1/2 and p52) expression and reducing viral gene (UL123, UL44 and UL32) replication in the HCMV-infected WI-38 cells. At viral attachment stage, Ts1-1A interacted with HCMV and prevented HCMV from attaching to its host cells. While at early phase of viral replication stage, Ts1-1A suppressed HCMV replication by downregulating NQO1 and HO-1 proteins related to oxidative stress as an antioxidant. To sum up, Ts1-1A is a promising anti-HCMV agent which could be developed for HCMV infection prevention and therapy.

Activity of Lignin-Modifying Enzyme of Selected Medicinal Mushrooms in Submerged Fermentation of Lignocellulosic Materials.

In the present study, wide diversity in the set and activity of lignin-modifying enzymes (LME) was revealed during submerged fermentation of mandarin peel with 15 strains of white rot Basidiomycetes. Among them, Trametes pubescens BCC153 was distinguished by the simultaneous production of laccase, manganese peroxidase (MnP), and lignin peroxidase (LiP). Supplementation of CuSO4 at a concentration of 1 mM in the media for the cultivation of four Trametes species manifold increased the production of laccase. The diverse effects of chemically different lignocellulosic growth substrates and nitrogen sources on the production of individual LME have been established. The maximum laccase activity of T. pubescens was observed when the fungus was cultivated on media containing mandarin peel and wheat bran, whereas the highest MnP and LiP activities were detected in the submerged fermentation of tobacco residue. Peptone and casein hydrolysate appeared to be the best sources of nitrogen to produce laccase and both peroxidases by T. pubescens BCC153 whereas KNO3 was the worst nitrogen-containing compound for the production of all enzymes.

Production of a Cheese-Like Aroma via Fermentation of Plant Proteins and Coconut Oil with the Basidiomycetes Cyclocybe aegerita and Trametes versicolor.

Cheese is one of the most common dairy products and is characterized by its complex aroma. However, in times of climate change and resource scarcity, the possibility to mimic the characteristic cheese-like aroma from plant-based sources is in demand to offer alternatives to cheese. Accordingly, the production of a natural cheese-like aroma via fermentation of four plant-based proteins and coconut oil with basidiomycetes has been addressed. Mixtures of soy and sunflower protein with coconut oil (15 g/L) have shown the formation of a cheese-like aroma after 72 and 56 h after fermentation with Cyclocybe aegerita and Trametes versicolor, respectively. Isovaleric acid, butanoic acid, ethyl butanoate, 1-octen-3-ol, and various ketones were identified as the key odorants. Similarities to typical cheeses were observed by the principal component analysis. Overall, the finding offered an approach to a sustainable production of a natural cheese-like aroma from a plant source, thus contributing to the development of cheese alternatives.

Decolorization and detoxication of malachite green by engineered Saccharomyces cerevisiae expressing novel thermostable laccase from Trametes trogii.

Malachite Green (MG) is a widely used industrial dye that is hazardous to health. Herein, the decolourisation and detoxification of MG were achieved using the engineered Saccharomyces cerevisiae expressing novel thermostable laccase lcc1 from Trametes trogii. The engineered strain RCL produced a high laccase activity of 121.83 U L-1. Lcc1 was stable at temperatures ranging from 20 ℃ to 60 ℃ and showed a high tolerance to organic solvents. Moreover, Lcc1 could decolorize different kinds of dyes (azo, anthraquinone and triphenylmethane), among which, the decolorization ability of MG is the highest, reaching 95.10 %, and the decolorization rate of other triphenylmethane dyes also over 50 %. The RCL decolorized about 95 % of 50 mg L-1 of MG dye in 10 h at 30 ℃. The MG degradation products were analyzed. The industrial application potential of the RCL was evaluated by treating industrial wastewater and the decolourisation rates were over 90 %.

Identification of redox activators for continuous reactivation of glyoxal oxidase from Trametes versicolor in a two-enzyme reaction cascade.

Glyoxal oxidases, belonging to the group of copper radical oxidases (CROs), oxidize aldehydes to carboxylic acids, while reducing O2 to H2O2. Their activity on furan derivatives like 5-hydroxymethylfurfural (HMF) makes these enzymes promising biocatalysts for the environmentally friendly synthesis of the bioplastics precursor 2,5-furandicarboxylic acid (FDCA). However, glyoxal oxidases suffer from inactivation, which requires the identification of suitable redox activators for efficient substrate conversion. Furthermore, only a few glyoxal oxidases have been expressed and characterized so far. Here, we report on a new glyoxal oxidase from Trametes versicolor (TvGLOX) that was expressed at high levels in Pichia pastoris (reclassified as Komagataella phaffii). TvGLOX was found to catalyze the oxidation of aldehyde groups in glyoxylic acid, methyl glyoxal, HMF, 2,5-diformylfuran (DFF) and 5-formyl-2-furancarboxylic acid (FFCA), but barely accepted alcohol groups as in 5-hydroxymethyl-2-furancarboxylic acid (HMFCA), preventing formation of FDCA from HMF. Various redox activators were tested for TvGLOX reactivation during catalyzed reactions. Among them, a combination of horseradish peroxidase and its substrate 2,2'-azino-di-(3-ethylbenzthiazoline sulfonic acid) (ABTS) most efficiently reactivated TvGLOX. Through continuous reactivation of TvGLOX in a two-enzyme system employing a recombinant Moesziomyces antarcticus aryl-alcohol oxidase (MaAAO) almost complete conversion of 8 mM HMF to FDCA was achieved within 24 h.

Enhancement of the biological activity of hydroxytyrosol through its oxidation by laccase from Trametes versicolor.

The laccase-catalyzed oxidation of hydroxytyrosol (HT) towards the formation of its bioactive oligomer derivatives was investigated. The biocatalytic oligomerization was catalyzed by laccase from Trametes versicolor in aqueous or various water-miscible organic solvents and deep eutectic solvent (DES)-based media. Mass Spectroscopy and Nuclear Magnetic Resonance were used for the characterization of the products. The solvent system used significantly affects the degree of HT oligomerization. The use of 50 % v/v methanol favored the production of the HT dimer, while other organic solvents as well as DESs led to the formation of hydroxytyrosol trimer and other oligomers. In vitro studies showed that the HT dimer exhibits 3- to 4-fold enhanced antibacterial activity against Gram-positive and Gram-negative bacteria compared to the parent compound. Moreover, the ability of HT dimer to inhibit the activity of soybean lipoxygenase and Candida rugosa lipase was 1.5-fold higher than HT, while molecular docking supported these results. Furthermore, HT dimer showed reduced cytotoxicity against HEK293 cells and exhibited a strong ability to inhibit ROS formation. The enhanced bioactivity of HT dimer indicates that this compound could be considered for use in cosmetics, skin-care products, and nutraceuticals.

In-vivo and in-vitro wound healing and tissue repair effect of Trametes versicolor polysaccharide extract.

Regarding different medical benefits of fungi, using the medical mushroom extracts as wound-healing agents is gaining popularity. This study, evaluated the wound healing characteristics of Trametes versicolor. Anti-oxidant activity addressed by employing the DPPH (2,2-diphenyl-1-picrylhydrazyl) assay resulting 53.7% inhibitory effect. Besides, for anti-microbial ability determination, the MIC (Minimum Inhibitory Concentration) of extract measured which Escherichia coli growth was inhibited at 1.1 mg/ml, and Staphylococcus aureus did not grow at 4.38 mg/ml of extract. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method indicated dose dependence of the extract with 63 ± 3% and 28 ± 3% viability at 1250 µg/ml and 156.25 µg/ml of extract, which higher concentration caused higher cell viability. The outcome of gene expression analysis determined that overall expression of FGF2 (Fibroblast Growth Factor 2), IL-1ß (Interleukin-1ß), and TGF-ß1 (Transforming Growth Factor-ß1) was 4 times higher at 48 h than at 24 h in treated cells, suggesting a stimulating effect on cell growth. An in-vivo animal model suggested enhanced wound healing process after treatment with 0.01 g of extract. Furthermore, the number of fibroblasts, epidermal thickness, and collagen fiber was respectively 2, 3, and threefold higher in treated mice when compared to untreated mice. The treated wounds of mice showed 100% and 60% of untreated mice of healing within 14 days. The results of this research show promise for the fungus-based wound healing treatments, which may help with tissue regeneration and the healing of cutaneous wounds.

Antiproliferative and Antimigratory Activity of Poly-gallic Acid in Cancer Cell Lines.

BACKGROUND/AIM: Enzyme-mediated grafting of poly (gallic acid) (PGAL) and L-arginine and a-L-lysine onto PGAL produces reactive oxygen species (ROS)-suppressor multiradical molecules with low cytotoxicity, high thermostability and water solubility with cancer treatment potential. This study examined the anticancer effects of these molecules in hepatic (HepG2, ATCC HB-8065), breast (MCF7, ATCC HTB-22), and prostate (PC-3, ATCC CRL-1435 and DU 145, ATCC HTB-81) cancer cell lines, as well as in fibroblasts from healthy human skin as control cells. MATERIALS AND METHODS: PGAL was synthesized by the oxidative polymerization of the naturally abundant GA using laccase from Trametes versicolor. Insertions of amino acids L-arginine and α-L-lysine on the PGAL chain were carried out by microwave. The cells of dermal fibroblast (Fb) were obtained from primary skin cultures and isolated from skin biopsies. The cancer cells lines of hepatic (HepG2), breast (MCF7), and prostate (PC-3, DU 145) were obtained from ATCC. The viability of the cancer cells and the primary culture was obtained by the MTT assay. Proliferation was demonstrated by crystal violet assay. Cell migration was determined by Wound healing assay. Finally, cell cycle analysis was carried out with cells. RESULTS: The results show that 200 µg/ml of PGAL cultured in vitro with prostate cancer cells decreased viability, proliferation, and migration, as well as arrested cells in the G1 and S phases of the cell cycle. In contrast, the dermal fibroblasts and the hepatic line remained unaffected. The random grafting of L-Arg and a-L-Lys onto the PGAL chain also decreased the viability of prostate cancer cells. CONCLUSION: PGAL and PGAL-grafted amino acids are potential adjuvants for prostate cancer treatment, with improved physicochemical characteristics compared to GA.

Immobilization of recombinant Trametes versicolor aflatoxin B1-degrading enzyme (TV-AFB1D) with montmorillonite for absorption and in situ degradation of aflatoxin B1.

Aflatoxin B1 is a highly carcinogenic and teratogenic substance mainly produced by toxin-producing strains such as Aspergillus flavus and Aspergillus parasitic. The efficient decomposition of aflatoxin is an important means to reduce its harm to humans and livestock. In this study, Trametes versicolor aflatoxin B1-degrading enzyme (TV-AFB1D) was recombinantly expressed in Bacillus subtilis (B. subtilis) 168. MMT-CTAB-AFB1D complex was prepared by the immobilization of TV-AFB1D and montmorillonite (MMT) by cross-linking glutaraldehyde. The results indicated that TV-AFB1D could recombinantly express in engineered B. subtilis 168 with a size of approximately 77 kDa. The immobilization efficiency of MMT-CTAB-AFB1D reached 98.63% when the concentration of glutaraldehyde was 5% (v/v). The relative activity of TV-AFB1D decreased to 72.36% after reusing for 10 times. The content of AFB1 in MMT-CTAB-AFB1D-AFB1 decreased to 1.1 µg/g from the initial 5.6 µg/g after incubation at 50 °C for 6 h. The amount of 80.4% AFB1 in the MMT-CTAB-AFB1D-AFB1 complex was degraded by in situ catalytic degradation. Thus, the strategy of combining adsorption and in situ degradation could effectively reduce the content of AFB1 residue in the MMT-CTAB-AFB1D complex.

A novel, robust peptidyl-lys metalloendopeptidase from Trametes coccinea recombinantly expressed in Komagataella phaffii.

A novel peptidyl-lys metalloendopeptidase (Tc-LysN) from Tramates coccinea was recombinantly expressed in Komagataella phaffii using the native pro-protein sequence. The peptidase was secreted into the culture broth as zymogen (~38 kDa) and mature enzyme (~19.8 kDa) simultaneously. The mature Tc-LysN was purified to homogeneity with a single step anion-exchange chromatography at pH 7.2. N-terminal sequencing using TMTpro Zero and mass spectrometry of the mature Tc-LysN indicated that the pro-peptide was cleaved between the amino acid positions 184 and 185 at the Kex2 cleavage site present in the native pro-protein sequence. The pH optimum of Tc-LysN was determined to be 5.0 while it maintained ≥60% activity between pH values 4.5-7.5 and ≥30% activity between pH values 8.5-10.0, indicating its broad applicability. The temperature maximum of Tc-LysN was determined to be 60 °C. After 18 h of incubation at 80 °C, Tc-LysN still retained ~20% activity. Organic solvents such as methanol and acetonitrile, at concentrations as high as 40% (v/v), were found to enhance Tc-LysN's activity up to ~100% and ~50%, respectively. Tc-LysN's thermostability, ability to withstand up to 8 M urea, tolerance to high concentrations of organic solvents, and an acidic pH optimum make it a viable candidate to be employed in proteomics workflows in which alkaline conditions might pose a challenge. The nano-LC-MS/MS analysis revealed bovine serum albumin (BSA)'s sequence coverage of 84% using Tc-LysN which was comparable to the sequence coverage of 90% by trypsin peptides. KEY POINTS: •A novel LysN from Trametes coccinea (Tc-LysN) was expressed in Komagataella phaffii and purified to homogeneity •Tc-LysN is thermostable, applicable over a broad pH range, and tolerates high concentrations of denaturants •Tc-LysN was successfully applied for protein digestion and mass spectrometry fingerprinting.

Huaier Regulates Oxaliplatin Resistance in Colorectal Cancer by Regulating Autophagy and Inhibiting the Wnt/ß-catenin Signalling Pathway.

OBJECTIVE: The present study aims to investigate the effect of Huaier on oxaliplatin (OXA) resistance in HCT-8 colorectal cancer (CRC) cells. METHODS: Oxaliplatin-resistant HCT-8/L CRC cells were used. The Cell Counting Kit-8, western blotting, quantitative real-time polymerase chain reaction, protein extraction kit, immunofluorescence and acridine orange staining assays were used in the study. The experiment results proved that Huaier has an influence on the Wnt/ß-catenin signalling pathway, autophagy and drug resistance. The authors of the present study used chloroquine, an autophagy inhibitor and Wnt agonist 1 (a Wnt pathway agonist) to verify the present experiment. RESULTS: The results showed that Huaier can regulate autophagy, inhibit the Wnt/ß-catenin signalling pathway and reverse the drug resistance of OXA-resistant CRC cells. CONCLUSIONS: This study proved that Huaier can regulate autophagy, inhibit the Wnt/ß-catenin signalling pathway and reverse the drug resistance of OXA-resistant CRC cells.

New insights in the biodegradation of high-cyclic polycyclic aromatic hydrocarbons with crude enzymes of Trametes versicolor.

High-cyclic polycyclic aromatic hydrocarbons (PAHs), with complex fused aromatic structures, are widespread, refractory and harmful in soil, but the current remediation technologies for high-cyclic PAHs are often inefficient and costly. This study focused on the biodegradation process of high-cyclic benzo[a]pyrene by Trametes versicolor crude enzymes. The crude enzymes exhibited high laccase activity (22112 U/L) and benzo[a]pyrene degradation efficiency (42.21%) within a short reaction time. Through the actual degradation and degradation kinetics, the degradation efficiency of PAHs decreased with the increase of aromatic rings. And the degradation conditions (temperature, pH, Cu2+ concentration, mediator) were systematically optimised. The optimum degradation conditions (1.5 mM Cu2+, 28℃ and pH 6) showed significant degradation efficiency for the low and medium concentrations of benzo[a]pyrene. In addition, complete degradation of benzo[a]pyrene could be achieved using only 0.2 mM of HBT mediator compared with crude enzymes alone. Collectively, these results showed the high-cyclic PAHs degradation potential of Trametes versicolor crude enzymes, and provided references to evaluate applicable prospects of white rot fungus crude enzymes in PAHs-contaminated soils.

Huaier improves the efficacy of anti-PD-L1 Ab in the treatment of hepatocellular carcinoma by regulating tumor immune microenvironment.

BACKGROUND: Combination therapy is an effective method for augmenting the efficacy of immune checkpoint inhibitors (ICIs). Huaier is a commonly used Chinese patent medicine with substantial antitumor effects. The combination of Huaier and ICIs may increase the efficacy of ICIs against hepatocellular carcinoma (HCC). METHODS: The major components of Huaier were detected by high-performance liquid chromatography-mass spectrometry. The optimal antitumor dose of Huaier was investigated in H22-bearing mice. Next, Huaier was combined with anti-CD8α antibody (Ab) or anti-PD-L1 Ab to observe the antitumor effect. The safety of these combination drugs was evaluated through blood biochemical tests and hematoxylin and eosin staining of histological sections. RT-qPCR, immunohistochemistry, flow cytometry, and transcriptome sequencing were performed to investigate the potential action mechanism of anti-PD-L1 Ab combined with Huaier against HCC. RESULTS: HPLC-MS/MS identified 333 components of Huaier, including carboxylic acids and derivatives, thienothiophenes, phenols, flavonoids and so on. Huaier exhibited significant antitumor effects, with the strongest effect noted at a dose of 4 g/kg. Huaier boosted CD8+ T cells infiltration into the tumor. Next, CD8+ T cells were depleted by with anti-CD8α Ab, and the antitumor effect of Huaier was suppressed. Flow cytometry results revealed that CD8+ T cells were reduced in the Huaier+anti-CD8α Ab group, with the antitumor effect of this group being inhibited. This indicated that CD8+ T cells were key players in the antitumor activity of Huaier. Meanwhile, Huaier inhibited microvessel density (MVD), downregulated vascular endothelial growth factor A (VEGFA), and upregulated PD-L1 in tumor tissues. Finally, Huaier combined with anti-PD-L1 Ab exhibited a greater antitumor effect in the H22-bearing mice. And the results of liver and kidney function tests and histological section analysis unveiled that the safety of these drugs was excellent. According to the transcriptome sequencing results, Huaier combined with anti-PD-L1 Ab possibly exerted anti-HCC effects through immunomodulation, immune response, and so on. CONCLUSIONS: Huaier exhibited a significant antitumor effect. It promoted CD8+ T cells infiltration, upregulated PD-L1 expression, downregulated VEGFA expression, and inhibited MVD, thereby playing a significant antitumor immunoregulatory effect. The combination of Huaier and anti-PD-L1 Ab has significant antitumor effects, and this regimen has good safety. Therefore, Huaier combined with anti-PD-L1 Ab is a promising therapeutic approach against HCC.

Biotransformation of metenolone acetate and epiandrosterone by fungi and evaluation of resulting metabolites for aromatase inhibition.

The present study describes the microbial transformation of anabolic drugs, metenolone acetate (1), and epiandrosterone (6). Three new metabolites, 6ß,17ß-dihydroxy-1-methyl-3-oxo-5α-androst-1-en (2), 5α,15α-dihydroxy-1-methyl-3-oxo-1-en-17-yl acetate (3), 15ß-hydroxy-1-methyl-3-oxo-5α-androst-1,4-dien-17-yl acetate (4), and a known metabolite, 17ß-hydroxy-1-methyl-4-androstadiene-3-one (5) were obtained by biotransformation of metenolone acetate (1) via Trametes hirsuta mushroom. Metabolites 7, and 8 were obtained from the incubation of epiandrosterone (6) with Cunninghamella blakesleeana. While bioconversion of compound 6 with Aspergillus alliaceus yielded seven known metabolites 9-15. Modern spectroscopic techniques were employed for the structure elucidation of biotransformed products. All compounds were evaluated for their aromatase inhibitory activity. Among them, new metabolite 3 exhibited a significant human placental aromatase activity with an IC50 = 19.602 ± 0.47 µM, as compared to standard anti-cancer drug exemestane (IC50 = 0.232 ± 0.031 µM), whereas, metabolite 5 (IC50 = 0.0049 ± 0.0032 µM) exhibited a very potent activity. While substrate 6, and metabolites 2, 7, and 9 were found inactive. Aromatase plays a key role in the biosynthesis of estrogen hormone, responsible for cancer cell proliferation. Its inhibition is therefore targeted for the treatment of ER + breast cancer. Further structural modifications (lead optimization) of compound 3 can lead to more potent aromatase inhibition for possible treatment of ER + breast cancer.

The Importance of Traditional Chinese Medicine in the Intervention and Treatment of HIV while Considering its Safety and Efficacy.

Natural products have been considered a potential resource for the development of novel therapeutic agents, since time immemorial. It is an opportunity to discover cost-effective and safe drugs at the earliest, with the goal to hit specific targets in the HIV life cycle. Natural products with inhibitory activity against human immunodeficiency virus are terpenes, coumarins, flavonoids, curcumin, proteins, such as lectins, laccases, bromotyrosines, and ribosome-inactivating proteins. Terpenes inhibit virus fusion, lectins and flavonoids have an inhibitory impact on viral binding, curcumin and flavonoids inhibit viral DNA integration. The most important medicinal plants which have been used in traditional Chinese medicinal sciences with anti-HIV properties are Convallaria majalis, Digitalis lanata, Cassia fistula, Croton macrostachyus, Dodonaea angustifolia, Ganoderma lucidum, Trametes versicolor, Coriolus versicolor, Cordyceps sinensis, Gardenia jasminoides, Morus alba, Scutellaria baicalensis, Ophiopogon japonicus, Platycodon grandiflorus, Fritillaria thunbergii, Anemarrhena asphodeloides, Trichosanthes kirilowii, Citrus reticulata, Glycyrrhiza uralensis, Rheum officinale, Poria cocos, Rheum palmatum, Astragalus membranaceus, Morinda citrifolia, Potentilla kleiniana, Artemisia capillaris, Sargassum fusiforme, Piperis longi fructus, Stellera chamaejasme, Curcumae rhizoma, Dalbergia odorifera lignum, Arisaematis Rhizoma preparatum, and Phellodendron amurense. The information provided is gathered from randomized control experiments, review articles, and analytical studies and observations, which are obtained from different literature sources, such as Scopus, Google Scholar, PubMed, and Science Direct from July 2000 to August 2023. The aim of this review article is to survey and introduce important medicinal plants and herbs that have been used for the treatment of HIV, especially the medicinal plants that are common in traditional Chinese medicine, as research to date is limited, and more evidence is required to confirm TCM,s efficacy.

Durability of heat-treated Paulownia tomentosa and Pinus koraiensis woods in palm oil and air against brown- and white-rot fungi.

This study aimed to evaluate and compare the effects of oil- and air-heat treatments on the durability of Paulownia tomentosa and Pinus koraiensis woods against Fomitopsis palustris and Trametes versicolor. The wood samples were treated in palm oil and air at 180, 200, and 220 °C for 2 h. The weight loss, morphology, crystalline properties, and chemical compounds of untreated and heat-treated wood after fungal attack were investigated. The significant difference in weight loss between oil- and air-heat-treated samples was shown at 220 °C. Heat-treated wood exposed to white-rot fungus showed a lower weight loss than that exposed to brown-rot fungus. The cell components in the untreated- and heat-treated Paulownia tomentosa and Pinus koraiensis at 180 °C were severely damaged due to fungal exposure compared to those at 220 °C. A fungal effect on the relative crystallinity was observed in heat-treated wood at 180 °C, whereas the effect was not observed at 220 °C. Following brown-rot fungus exposure, untreated- and heat-treated wood at 180 °C showed a notable change in the Fourier transform infrared (FTIR) peaks of polysaccharides, whereas no noticeable change in lignin peaks was observed. Heat-treated wood at 220 °C showed no noticeable change in the FTIR spectra owing to brown-rot fungus exposure. Exposure to white-rot fungus did not noticeably change the FTIR spectra of untreated and heat-treated wood.