Submerged Culture of Edible and Medicinal Mushroom Mycelia and Their Applications in Food Products: A Review.

Edible mushrooms have rich nutrition (e.g., proteins, dietary fibers, polysaccharides) and they can be potential sources of important ingredients in food processing. However, the cultivation of mushroom fruiting bodies needs a relatively long time, and they can be easily polluted during the growth process. At the same time, a lot of labor and larger planting areas are also required. As we all know, submerged fermentation is a good way to produce edible mushroom mycelia with less environmental pollution and small footprint, which are also rich in nutrition and bioactive components that are used as dietary supplements or health care products in the food industry. Therefore, it can be considered that the replacement of edible mushroom fruiting bodies with edible mushroom mycelia produced through submerged fermentation has great application potential in food production. At present, most of the research about edible mushroom mycelia focuses on the production of bioactive metabolites in fermentation liquid, but there are few reports that concentrate on their applications in food. This paper reviews the research progress of submerged culture of edible mushroom mycelia and their applications in food products.

A Synthetic Small RNA Homologous to the D-Loop Transcript of mtDNA Enhances Mitochondrial Bioenergetics.

Mitochondrial malfunction is a hallmark of many diseases, including neurodegenerative disorders, cardiovascular and lung diseases, and cancers. We previously found that alveolar progenitor cells, which are more resistant to cigarette smoke-induced injury than the other cells of the lung parenchyma, upregulate the mtDNA-encoded small non-coding RNA mito-ncR-805 after exposure to smoke. The mito-ncR-805 acts as a retrograde signal between the mitochondria and the nucleus. Here, we identified a region of mito-ncR-805 that is conserved in the mammalian mitochondrial genomes and generated shorter versions of mouse and human transcripts (mmu-CR805 and hsa-LDL1, respectively), which differ in a few nucleotides and which we refer to as the "functional bit". Overexpression of mouse and human functional bits in either the mouse or the human lung epithelial cells led to an increase in the activity of the Krebs cycle and oxidative phosphorylation, stabilized the mitochondrial potential, conferred faster cell division, and lowered the levels of proapoptotic pseudokinase, TRIB3. Both oligos, mmu-CR805 and hsa-LDL1 conferred cross-species beneficial effects. Our data indicate a high degree of evolutionary conservation of retrograde signaling via a functional bit of the D-loop transcript, mito-ncR-805, in the mammals. This emphasizes the importance of the pathway and suggests a potential to develop this functional bit into a therapeutic agent that enhances mitochondrial bioenergetics.

Flavin Adenine Dinucleotide Depletion Caused by electron transfer flavoprotein subunit alpha Haploinsufficiency Leads to Hepatic Steatosis and Injury in Zebrafish.

The electron transfer flavoprotein (ETF) complex, made up of the ETF alpha subunit (ETFA), ETF beta subunit (ETFB), and ETF dehydrogenase (ETFDH), regulates fatty acid ß-oxidation activity while scavenging leaked electrons through flavin adenine dinucleotide (FAD)/reduced form FAD (FADH2) redox reactions in mitochondria. Here, we hypothesized that ETF dysfunction-mediated FAD deficiency may result in increased mitochondrial oxidative stress and steatosis and subsequent liver injury. We report that etfa haploinsufficiency caused hyperlipidemia, hypercholesterolemia, and hepatic steatosis and injury in adult zebrafish. Further, etfa+/ - mutant livers had reduced levels of FAD and glutathione and an increase in reactive oxygen species. Because FAD depletion might be critical in the pathogenesis of the liver lesion identified in etfa+/ - mutants, we used riboflavin to elevate FAD levels in the liver and found that riboflavin supplementation significantly suppressed hepatic steatosis and injury in etfa+/ - mutants through suppression of oxidative stress and de novo lipogenesis in the liver. Additionally, we found that adenosine triphosphate-linked mitochondrial oxygen consumption and mitochondrial membrane potential were reduced in etfa+/ - primary hepatocytes and that riboflavin supplementation corrected these defects. Conclusion: FAD depletion caused by etfa haploinsufficiency plays a key role in hepatic steatosis and oxidative stress-mediated hepatic injury in adult zebrafish. This raises the possibility that people with ETFA haploinsufficiency have a high risk for developing liver disease.

Enhancement of carotenoid production and its regulation in edible mushroom Cordyceps militaris by abiotic stresses.

Cordyceps militaris carotenoids are widely used as food additives, animal feed supplements, and so on. However, the biosynthetic pathway of carotenoids in C. militaris is still obscure. In this paper, changes of mycelial morphology and carotenoid accumulation of C. militaris were investigated under oxidative (KMnO4) and osmotic stress (NaCl). Subsequently, qRT-PCR was employed to detect the expression levels of genes related to carotenogenesis to explore the mechanism of adaptation to abiotic stress. When the concentrations of KMnO4 and NaCl were respectively 0.4 g/L and 2 g/L, carotenoid accumulation reached a maximum of 6616.82 ±â€¯666.43 µg/g and 6416.77 ±â€¯537.02 µg/g. Under the oxidative stress condition of KMnO4, the expressions of psy and hsp70 increased significantly compared with control. Besides, the genes fus3 and hog1 were significantly enriched in the MAPK signal pathway. Compared with the control group, there was no significant difference in expression of psy in the NaCl group. Moreover, the accumulation of triacylglycerols may contribute significantly to the increase in carotenoid accumulation. The increased accumulation of antioxidant carotenoids induced under environmental stress is to resist oxidative conditions. Fus3 and Hog1 signaling in the MAPK pathway was activated and subsequently take effects on the resistance of oxidative condition by regulating related metabolic processes. C. militaris resist the stress of high oxygen by producing a large amount of glycerol and carotenoids when this fungus is cultured in a saline environment for a long time.

An Efficient Strategy for Enhancement of Bioactive Compounds in the Fruit Body of Caterpillar Medicinal Mushroom, Cordyceps militaris (Ascomycetes), by Spraying Biotic Elicitors.

Cordyceps militaris is a mushroom species with high nutritive and medicinal values based on diverse bioactive metabolites. The contents of bioactive ingredients are indicative of the quality of commercially available fruit body of this fungus. Although the application of biotic elicitors has been an efficient strategy to induce the accumulation of valuable bioactive compounds in vivo, related research in C. militaris is rarely reported. In this study, five biotic elicitors in different concentrations (0.05, 0.5, 1, and 2 mg/mL), including chitosan (CHT), 2,4-dichlorophenoxyacetic acid (2,4-D), methyl jasmonate (MeJA), gibberellic acid (GA), and triacontanol (TRIA), were first introduced to enhance the production of 10 kinds of major bioactive components in the fruit body of C. militaris. Results showed that the effect of biotic elicitors on bioactive compounds in the fruit body of C. militaris was elicitor-specific and concentration-dependent. Overall, 1 mg/L CHT was considered the most favorable for the production of 10 bioactive ingredients in C. militaris fruit body, which could increase the content of protein, polysaccharides, polyphenol, triterpenoids, flavonoids, cordyceps acid, cordycepin, and anthocyanins by 20.38-, 1.41-, 0.7-, 0.47-, 11.90-, 1.09-, 0.34-, and 2.64-fold, respectively, compared with the control. The results of this study would provide an efficient strategy for the production of a superior quality fruit body of and contribute to further elucidation of the effects of biotic elicitors on metabolite accumulation in C. militaris.

Transcriptome Analysis of Cordyceps militaris Reveals Genes Associated With Carotenoid Synthesis and Identification of the Function of the Cmtns Gene.

Cordyceps militaris, a valuable edible and medicinal fungus, has attracted increasing attention because of its various bioactive ingredients. However, the biosynthetic pathway of C. militaris carotenoids is still unknown due to lack of transcriptome information. To uncover genes related to the biosynthesis of C. militaris carotenoids, the transcriptomes of mycelia CM10_D cultured under dark conditions and mycelia CM10_L cultured under light exposure conditions were sequenced. Compared with mycelia CM10_D, 866 up-regulated genes and 856 down-regulated genes were found in mycelia CM10_L. Gene ontology (GO) analysis of differentially expressed genes (DEGs) indicated that DEGs were mainly classified into the "metabolic process," "membrane," and "catalytic activity" terms. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of DEGs suggested that DEGs were mainly enriched in "metabolic pathways," "MAPK signaling pathway-yeast," and "biosynthesis of secondary metabolites." In addition, the carotenoid content of the Cmtns gene deletion mutant (ΔCmtns) was significantly lower than that of the wild-type C. militaris CM10, while the carotenoid content of the complementary strain (ΔCmtns-c) of the Cmtns gene was not significantly different from that of C. militaris CM10, suggesting that the Cmtns gene significantly affected the biosynthesis of carotenoids in C. militaris. These results potentially pave the way for revealing the biosynthetic pathway of carotenoids and improving carotenoids production in C. militaris.

In-vivo pharmacokinetics, tissue distribution and anti-tumour effect of hydroxycamptothecin delivered in oil-in-water submicron emulsions.

OBJECTIVES: The aim of this study was to investigate the pharmacokinetics, tissue distribution and anti-tumour effect of hydroxycamptothecin submicron emulsions (HCPT-SEs). METHODS: HCPT-SEs or HCPT injection (HCPT-I) was administered intravenously into the tail vein of rats or S180 tumour-bearing mice. KEY FINDINGS: HCPT-SEs increased the plasma concentration of HCPT compared with HCPT-I at all time points. The AUC(0-∞) , elimination half-life and mean residence time of anionic submicron emulsions containing HCPT (HCPT-ASEs) and cationic submicron emulsions containing HCPT (HCPT-CSEs) were significantly greater than those of HCPT-I (P <0.01). Especially, a prolonged elimination half-life was found for HCPT-CSEs. HCPT-CSEs and HCPT-ASEs resulted in a 7.9-fold and 3.1-fold increase in AUC(0-6h) of tumour compared with HCPT-I, respectively. The targeting efficiency (T(e) ) of HCPT-ASEs and HCPT-CSEs indicated their selectivity to tumour and the T(e) of HCPT-CSEs was significantly higher than that of HCPT-ASEs (P<0.01). The anti-tumour effect studies showed that HCPT-SEs improved the therapeutic efficiency of HCPT compared with HCPT-I. The percentage of tumour growth suppression rate of mice treated with HCPT-CSEs (2.0 mg HCPT eq./kg) increased 2.1 fold compared with that of HCPT-I. CONCLUSIONS: Submicron emulsions can alter the pharmacokinetic characteristics and tissue distribution of HCPT, and enhance tumour targeting and anti-tumour activity.

In silico analysis of phytoene synthase and its promoter reveals hints for regulation mechanisms of carotenogenesis in Duanliella bardawil.

MOTIVATION: Previous researches showed that phytoene synthase (Psy) from Dunaliella bardawil is the first regulatory point in carotenogenesis. We hypothesize certain interactions between the environmental stress factors and the regulatory sequences of Psy in D.bardawil (DbPsy). Consequently, LA PCR-based genomic walking approach was performed for isolation of psy promoter and terminator, respectively. The obtained nucleic acid sequences and the corresponding protein structure of DbPsy were analyzed and predicted using various bioinformatics tools. Finally, we presented some hints for the regulation mechanisms of DbPsy at the molecular level according to the computed results. RESULTS: LA PCR-based genomic walking results showed that the isolated sequences are the promoter and terminator of psy, correspondingly. Computational analysis demonstrated several candidate motifs of the promoter exhibiting hypothetic UV-B-, norglurzon- and salt-induced characteristics, as well as some typical domains universally discovered in promoter sequences, such as TATA-box, CCAAT-box and GATA-box, etc. Furthermore, the structure of Psy was also predicted and aligned along with many counterparts at the protein level. Low homology of N-terminus was found in D.bardawil, while a relatively conserved C-terminus was predicted to be involved in the catalytic activity and substrate recognization/binding. Phylogenic analysis classified the DbPsy into a cluster with other algae. These results implied that Psy may share similar regulation mechanisms among algae with respect to their C-termini; while the diversity in N-terminus among Psys, along with the predicted inducible motifs in psy promoter from D.bardawil, may confer the fine tuning differences between D.bardawil and other algae. CONCLUSION: By means of computer techniques, we found in D.barawali that two interesting conserved motifs of psy promoter may involve in UV-B, norglurzon and salt regulation correspondingly; and that the diversity of Psy protein mainly lies in the N-termini among algae. These results indicate some hints for regulation mechanisms of carotenogenesis in D.bradawil. CONTACT: jgjiang@scut.edu.cn.

[Experimental study of Gukangling Decoction combined with technetium [99Tc] methylene diphosphonate injection in treating osteoporotic rabbits].

OBJECTIVE: To evaluate the advantage of Gukangling Decoction (GKLD), a compound traditional Chinese herbal medicine, combined with technetium [(99)Tc] methylene diphosphonate injection ((99)Tc-MDP) in treating osteoporosis in rabbits. METHODS: A rabbit model of osteoporosis was established by intramuscular injection of dexamethasone (DX). Fifty-six rabbits were divided into 8 groups: Group A, B, C, D, E, F, G and H. Rabbits in groups A and B were intramuscularly injected normal saline as normal control, groups C and D were untreated groups, rabbits in group E were treated by (99)-MDP, rabbits in group F were treated by aminodiphosphate, rabbits in group G were treated by GKLD, and rabbits in group H were treated by (99)-MDP and GKLD. Rabbits in groups A and C were executed to demonstrate the establishment of the rabbit model of osteoporosis at the 8th week of experiment. Rabbits in the other six groups were executed after 16-week experiment (8-week treatment), and then bone structure and cell shape were observed by electron microscope, X-ray, CT and emission computed tomography (ECT). Bone density, biomechanical parameters, the levels of bone specific alkaline phosphatase (BALP) and bone Gla protein (BGP) were measured too. RESULTS: After 8-week of intramuscular injection of DX, the bone trabecula in group A were regular and showed normal configuration, while the bone trabecula in group C were sparse, ruptured and showed damaged form. The bone density and biomechanical parameters in group A were higher than those in group C, indicating that the rabbit model of osteoporosis was established successfully. At the 9th week of experiment, the results of cell pathology in group D showed that the bone trabeculas were sparse, ruptured, defected or had hollow section, but the bone trabeculas in group B were regular and dense. The bone trabeculas in groups H and E were restored, and were thicker than those in group D. The bone quality in groups H and E was better than group D significantly, the bone quality in group F was better than group G, and the bone quality in group G was better than group D slightly. CONCLUSION: GKLD combined with (99)-MDP had superiority in treating osteoporosis of rabbits as compared with the respective single therapy.