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Sparassis crispa which looks like the light yellow flower of cabbage, spontaneously shapes and grows from edge of the stump or the dead tree in autumn. The mycelium of the mushroom has the ability which extirpation of the various germs. Also, known as a key component, protein-bound polysaccharide (PBP) has potential medical effects like anticancer and immunity. In this study, we establish the conditions for extraction of PBP from cultivated Sparassis crispa mycelium. In order to increase the extraction yield of PBP from the mycelium, first, three well known extraction methods such as hot water, ultrasonic and microwave were used. And then extract using pretreatment methods such as ultra-fine pulverization(UFP; particle sizes are less than 100 §) and ultra high pressure(UHP; 100 Mpa). Second, the most important factors of PBP extracted by using different two solvent(1 N NaOH and H2O). The results showed that total content of PBP by using 1 N NaOH as a extraction solvent was increased from 1.92 to 42.16 mg/g than H2O. Namely, the obtained optimum conditions from 1 N NaOH extraction method of the PBP from Sparassis crispa mycelium were 150¡É for 60 minutes. In addition, the functional group of PBP was confirmed by using FT-IR, and images of extracted Sparassis crispa mycelium were observed by using SEM and TEM.
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Mushroom ¥â-glucan are increasingly being utilized to treat a wide variety of diseases. ¥â-glucan (SCG), purified 6-branched 1, 3-beta-D- glucan obtained from Sparassis crispa, exhibits diverse biological activities including immunomodulation and antitumor activites. However, the cellular and molecular mechanism underlying its therapeutic effect is poorly understood. During the evolution of neoplastic diseases, dendritic cell (DC) functions are usually attenuated, and this presents a problem to DC-based immunotherapies against cancer. DCs play pivotal roles in the initiation of T cell-mediated immune response, making them an attractive in immuno vaccines. In this study, we investigated the effects of SCG on immune functions of dendritic cells. SCG efficiently increased the maturation of TLR4+/+ DCs from C57BL/6 and C3H/HeN mice, but not TLR4-/- DCs from C3H/HeJ mice. Phenotypic maturation was confirmed by the elevated expressions of CD40, CD80, CD86, MHC-class I, and MHC- class II molecules, and functional maturation by increased in the gene expression of IL-12, IL-1¥â, TNF-¥á, and IFN-¥á/¥â, and decreased endocytosis. Also, SCG made DCs to activate allogenic T cells to proliferate and to produce cytokines such as IL-2 and IFN-¥ã. As a mechanism of action, SCG was able to activate mitogen-activated protein kinases, such as ERK, p38, and JNK. In addition, SCG activated nuclear transcription factor NF-¥êB p65/p50, which are signaling molecules down-stream of toll-like receptor 4 (TLR4). SCG activity was not affect by polymyxin B, a specific inhibitor of LPS. These results indicate that DCs appear to be a potential target for the immunomodulatory capacity of SCG and suggest the possible use of SCG in DC-based immunotherapies.
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Sparassis latifolia (formerly known as S. crispa) is a culinary-medicinal mushroom that has recently become popular in Korea, China, and Japan. S. latifolia is a good source of food and nutraceuticals, or dietary supplements, due to its rich flavor compounds and b-glucan content. Researches have particularly been done of the effect of b-glucan on anti-cancer and immuno-modulation effects using mice models and human trials. While, some researches are doing on the effect of brain function including neuroprotective effect of mushroom b-glucan. Thus it would be good for this mushroom to have much nutrients such as g-aminobutyric acid (GABA) as well as b-glucan. Biofortification is agricultural technology which can enhance some nutritions in crops. Most of all, supplying the fertilized soil or media nutrients to mushrooms is the most safe and effective way of biofortification. GABA, a non-essential and free amino acid, is one of the functional compounds. It helps increasing the cognitive function and improving the insomnia. Mushrooms have the GABA metabolism and it is related to TCA cycles which is an essential for growth. In this study, the enhancement of growth and GABA production in a medicinal and edible mushroom mycelia, S. latifolia, was hypothesized and amino acids were added as biofortification factors.
 S. latifolia mycelia (JF02-06) was provided by Jeonnam Forest Research Institute (Naju, Korea). To biofortify mushroom with two amino acids, one major approach can be used. Mycelia were pre-incubated on agar plate. Experiment for effect of amino acids was conducted with liquid media. Amino acid free medium was potato dextrose broth (PDB). Amino acids added were L-glutamic acid (PDBG media) and L-ornithine (PDBO media). The range of amino acids is 0.6-2.2% (w/v) and PDB media was used as negative control. The mycelial growth was measured by diameter of mycelia of solid media and dried weight of mycelia for liquid media. GABA and b-glucan content of dried mycelia were measured. GABA was extracted from dried mycelia with 0.1 N HCl and the extracts were freeze-dried. Freeze-dried extracts were derivatized with 1,4-dinitrofluorobenzen (DNFB). GABA was analyzed with HPLC equipped with a C18 column and gredient elution of mobile phase. b-Glucan content was measured using an assay kit from Megazyme. All results of this study were statistically analyzed using IBM SPSS Statistics version 23. The results were as follows:
 1) The statistical analysis was conducted one-way ANOVA, and the result was F(10)=3.672, p<0.01. The growth of mycelia was determined to be 0.9¡¾0.00 g/L in PDB media. And in PDBG and PDBO were 2.2¡¾0.16 g/L and 1.93¡¾0.34 g/L, respectively.
 2) The statistical analysis was conducted one-way ANOVA, and the result was F(10)=20.183, p<0.001. The GABA content was 21.3¡¾0.9 mg/100 g in PDB media. In PDBG 1.4% media, it was 115.4¡¾30.2 mg/100 g and it was the best of all. However, the PDBO media was not effective to increasing the GABA content in mycelia.
 3) The statistical analysis was conducted one-way ANOVA, and the result was F(10)=0.823, p>0.05. Amino acids have no effect on change of b-glucan content of mycelia. The b-glucan content were 39.7¡¾1.4 mg/100 mg in PDB media, 34.4¡¾0.2 mg/100 mg in PDBG 2.0% media and 35.2¡¾9.2 mg/100 mg in PDBO 1.4% media.
 Addition of glutamic acid and ornithine positively affected the growth of S. latifolia mycelia and glutamic acid also positively affected the GABA production in mycelia. The key enzyme in GABA biosynthesis is glutamic acid decarboxylase (GAD) in GABA-shunt, a short GABA metabolism. So it seems that the glutamic acid is decarboxylated into GABA by GAD through the GABA-shunt. No degradation of GABA was observed in the addition of glutamic acid. More metabolism study should be done to elucidate the GABA-shunt of S. latifolia. The results of this study can be applied to other mushrooms production, and the biofortification-based approach can be a valuable asset in mushroom industry.
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Sparassis crispa is an edible and medicinal mushroom, which commonly called the cauliflower mushroom. S. crispa is known to be a good source of food and nutraceuticals due to their rich flavors and ¥â-glucan contents as well as a good source of antiviral and anticancer compounds. However, there have been no report on the effect on hair growth stimulation and/or growth inhibition. C57BL/6 mice the most widely used lab mouse strain were used in this animal study due to their availability and the similar hair growth cycle to human hair. In this study, hair-removed five-year-old C57BL/6 mice were used to examine the effect of S. crispa extracts(70% ethanol) on hair growth stimulation and inhibition of the mice. The extracts and control compounds were topically treated once a day for 4 weeks. The hair growth was photographically and histologically examined during the total period of 4 weeks. Hair growth of the test group was a little inhibited compared to positive (3% minoxidil) and negative control groups (water and ethanol treated groups), respectively. The whole data showed little improvement of hair growth effect of S. crispa extracts but with no real statistical significance. These results suggest that S. crispa extract has little hair growth promoting activity and more works should be done to elucidate the inhibition mechanism.
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