• Journal of Life Science
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• v.30 no.4
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• pp.359-369
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• 2020

Although many studies on immune modulatory materials have used RAW 264.7 cells, few have used T cell-derived TK-1 cell lines. Moreover, although some studies have investigated the efficacy of plant-derived β-sitosterol, few have examined the immunomodulatory activity of its analogue, daucosterol. In this study, β-sitosterol and daucosterol were isolated from D. batatas and identified by nuclear magnetic resonance spectroscopy. To evaluate the immune-enhancing or inhibitory effects of the isolated phytosterols, the expression levels of the inflammatory response genes COX-2, TNF-α, IL-6, and iNOS were analyzed by RT-PCR. The relative expression levels of TNF-α and iNOS in RAW 264.7 cells were increased more than threefold with β-sitosterol treatment comparing to those of untreated control. In the case of TK-1 cells, the expression level of TNF-α was decreased and the expression level of iNOS was increased in a β-sitosterol concentration-dependent manner. The expression levels of COX-2, TNF-α, and IL-6 increased by approximately 0.7-1.2 times in RAW 264.7 cells treated with daucosterol compared to those of untreated control, but iNOS expression decreased by 0.8-0.18 times. In the case of daucosterol-treated TK-1 cells, the expression levels of TNF-α, IL-6, and iNOS were markedly reduced from those of TK-1 cells treated only with lipopolysaccaride. As a conclusion, β-sitosterol treatment increased TNF-α and iNOS expression levels in RAW 264.7 cells, thus exerting an immune- boosting effect. However, in TK-1 cells, iNOS expression increased while TNF-α expression decreased, indicating an immunosuppressive activity of β-sitosterol. Daucosterol appears to exert an immunosuppressive effect in both macrophages and T cell lines by inhibiting iNOS expression in RAW 264.7 cells and greatly inhibiting the expression of TNF-α, IL-6, and iNOS in TK-1 cells.

• Journal of Life Science
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• v.14 no.1
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• pp.104-109
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• 2004

lt is well known that some kinds of leaves contain a lot of antioxidants in them; however, little attention has been given to the study on the amount of antioxidants in them and the changes in the amount of antioxidants in them during their growth. Therefore, we examined the antioxidants in the leaves of persimon, moraceae, and trifoliage orange during their growth. The amount of total polyphenol tended to decrease during plants' growth, and each amount was 4.62 g/dried 100 g persimon leaf, 1.70 gloried 100 g moraceae leaf, and 0.91 g/dried 100 g trifoliage orange leaf in April. The amount of total polyphenol in persimon leaf was 2.7∼5.1 times higher than moraceae and trifoliage orange leaf. The amount of ascorbic acid also decreased during plants' growth, and each amount was 2.7∼6.0 mg/dried 100 g moraceae leaf, 5.3∼9.9 mg/dried 100 g persimon leaf, and 3.7∼6.9 mg/dried 100 g trifoliage orange leaf. Persimon leaf was found to contain higher amount of ascorbic acid than moraceae leaf or trifoliage orange leaf. The amount of glutathione tended to decrease during plants' growth, and each average amount was 35.7 mg/dried 100 g trifoliage orange leaf, 15.8 mg/dried 100 g moraceae leaf, and 2.3 mg/dried 100 g persimon leaf. Trifoliage orange leaf contained the highest amount of glutathione. $\beta$ -Carotene tended to increase during the growth, and each amount was 411.2 mg/dried 100 g moraceae leaf, 198.5 mg/dried 100 g persimon leaf, 144.1 mg/dried 100 g trifoliage orange leaf in September. $\alpha$ -Tocopherol also tended to increase during the growth, and each amount was 52.8 mg/dried 100 g trifoliage orange leaf, 48.6 mg/dried 100 g moraceae leaf, and 61.7 mg/dried 100 g persimon leaf in September.

• Applied Biological Chemistry
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• v.18 no.4
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• pp.194-202
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• 1975

This study was conducted to study the properties of the water-soluble natural chelating agents from garbage compost and activated sewage sludge responsible for Fe chelation, which is closely associated with the effectiveness in correcting iron chlorosis in plant. The water-soluble fraction of these materials was fractionated by menas of Sephadex gel filtration and the fractions of Fe chehates were traced by radioactive $^{59}Fe$. The fractions were examined by ultraviolet and infrared. spectroscopy and stability constants for Fe. The water-soluble fraction from garbage compost was separated by Sephadex G-25 into approximately four fractions. Most of the added $^{59}Fe$ was associated with fraction I, which appeared at the void volume. Further fractionation by Sephadex G-50 indicated that the molecular weight of water-soluble chelating agents is in the approximate range of 5000 to 10,000. The water-soluble fraction from activated sewage sludge gave six fractions by Sephadex G-25. Most of the added $^{59}Fe$ was found in the fraction I,II, and III, The molecular weights of most chelating agents associated with $^{59}Fe$ appeared to be less than 5,000 and those of fraction I that appeared at the void volume was in the range of 5,000 to 1,000. Discrepancy between radio activity count and UV absorption indicated the heterogeneity of the fractions obtained by Sephadex gel filtration. Ultraviolet absorption spectra of all fractions separated by Sephadex G-25 and containing chelating agents showed no differences. Fraction IV and V of sewage extract showed absorption maxima and shifting similar to nucleic acid components suggesting the presence of decomposition products of nucleic acid. Similarity fraction VI contained phenolic type amino acid groups. Fraction I of compost extract contained most of the added $^{59}Fe$ and showed weak but extra definite absorption in the 1230, and $1270cm^{-1}$ region, suggesting that extra oxygen groups in polyphenolic structure were probably involved in Fe chelation. In sewage extract, fraction I,II, and III in which most of the $^{59}Fe$ was found, showed strong definite polypeptide absorption in the region of $1540cm^{-1}$ due to NH deformation and C-N stretching of amide groups in the peptidebond. These extra functional groups in fraction I, II, and III appeared to be associated with Fe chelation. The other fractions, not associated with $^{59}Fe$, still have carboxyl and hydroxyl groups, suggesting that these functional groups in these water extracts may not independently form the Fe chelates. Precipitation of ferric hydroxide precluded measuring the stability constants for Fe-chelates. However, the formation constants for Zn chelates as log K values for compost extract and sewage extract at pH 4.0 from which the strength of chelation with Fe could be presumed, were 8.23, and 9.75, respectively, indicating strong complexation with metals. The chelating capacity of compost extract containing 6.5 g organic matter per liter was 0.82 mM, and that of sewage extract containing 5.3 g per liter was 0. 64 mM.

• Applied Biological Chemistry
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• v.42 no.2
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• pp.116-122
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• 1999

In order to characterize the property of $Ca^{2+}$ transport in plant cell, microsomes were prepared from the roots of tomato and microsomal $^{45}Ca^{2+}$ uptake was measured. When 1 mM vanadate, a selective inhibitor of P-type ATPases, 50 mM $NO_3^-$, a specific inhibitor of vacuolar $H^{+}-ATPase$, and both of these inhibitors were treated, the microsomal $^{45}Ca^{2+}$ uptakes were inhibited by 20, 33 and 47%, respectively. The inhibitory effects of these two inhibitors were investigated by using a protonophore, gramicidin. When the chemical gradient of $H^{+}$ was relieved by gramicidin, the uptake was decreased by 30%, implying the presence of $Ca^{2+}/H^+$ antiporter in the microsomal membrane. In the $^{45}Ca^{2+}$ uptake experiment, the effect of gramicidin was independent of vanadate-induced inhibition. However, when the activity of vacuolar $H^{+}-ATPase$ was inhibited by $NO_3^-$, the effect of gramicidin was severely decreased. Meanwhile, thapsigargin, a specific antagonist of ER/SR-type $Ca^{2+}-ATPase$, inhibited the microsomal $^{45}Ca^{2+}$ uptake and the maximum inhibitory effect was obtained at $10\;{\mu}M$. The effect of thapsigargin was blocked by $NO_3^-$ and gramicidin, but not by vanadate. These results imply that vanadate directly inhibits the activity of $Ca^{2+}-ATPase$; however, $NO_3^-$ and thapsigargin block the activity of $Ca^{2+}/H^+$ antiporter by inhibiting the vacuolar $H^{+}-ATPase$. In conclusion, the microsomal $^{45}Ca^{2+}$ uptakes are mediated by two major enzymes, $Ca^{2+}-ATPase$ and $Ca^{2+}/H^+$ antiporter in tomato root tissue.

• Applied Biological Chemistry
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• v.8
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• pp.101-111
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• 1967

Daily doses of phenylmercuric acetate arranged in $30{\gamma}\;(group\;I)$, 3{\gamma}\;(group\;II)$ and $0.3{\gamma}\;(group\;III)$ were administered respectively to rabbits for 90 days. The chief histopathological changes in the organs and the analytical data on mercury residues in the excretion and liver were as follows. 1. Kidney: In group I, severe degrees of vacuolization and cloudy swelling were occurred in the epithelial cells of proximal convoluted tubules and severe cloudy swelling and coagulative necrosis were observed in the proximal straight tubules. There were many hyaline casts in the collecting tubules. In group II, moderate degrees of vacuolization and cloudy swelling were observed in the epithelial cells of proximal convoluted tubules and moderate cloudy swelling and coagulative necrosis were encountered in the proximal straight tubules. A little numbers of hyaline casts were located in the lumen of collecting tubules. In group III, slight degree of cloudy swelling were observed in the epithelial cells of proximal convoluted and straight tubules. 2. Liver: In group I, cloudy swelling, fatty changes and coagulative necrosis were observed in the central zone of hepatic lobules. Dissociation of hepatic cell cords was encountered. Hyperplsia of hepatic cells were remarkable in group II. No Pathological changes were observed in group III. 3. Spleen: Deposition of hemosiderin pigment was prominant in group I and small amount of the pigment was observed in group II. There were no pathological changes in group III. 4. Adrenal, colon and heart: No pathological changes were detected in all 3 groups. 5. In an average about 76.5% of mercury was excreted from group I, 85.4% from group II and 79.8% from group III. 6. Mercury content in the liver was 0.0348 g in group I, 0.00378 g and 0.00066 g in group II and group III respectively. 7. In general, as to increased mercury doses the concentration of mercury accumulation in the liver became higher, how·ever, the accumulation quantity against a total amount of mercury doses showed an adverse trend. In other word, the quantity of mercury accumulation was not increased proportionately by higher dose of mercury.

• Journal of Mushroom
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• v.9 no.2
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• pp.59-62
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• 2011

To develop a new cultivar of King oyster mushroom(Pleurotus eryngii), G09-21 as parental strain was selected by the method of Di-mon crossing between monokaryotic strains derived from ASI 2824(Keunneutari No.2) and dikaryotic strain ASI 2887(Aeryni 3). The Pe21-51($G09-21-10{\times}2844-11$) was shown the best cultural characteristics, selected to be a new cultivar and named as 'Song-A'. The 'Song-A' was formed incompatibility line distinctly in the confrontation growth of parental strains Keunneutari No.2, Aeryni 3 and ASI 2844. The optimum temperature for mycelial growth, fruiting body development and pH arrange were $25{\sim}30^{\circ}C$, $14{\sim}16^{\circ}C$ and pH5~8, respectively. Fruiting body production per bottle was about $94.7{\pm}29.5$ g which is almost 106% quantity compared to that of other cultuvar Keunneutari No.2. And also the stip is thick and long but the number of available stipe is few. Analysis of the genetic characteristics of the new cultivar 'Song-A' showed a different DNA profile as that of the control strains, Keunneutari No.2, Aeryni 3 and ASI 2844, when RAPD(Random Amplified Polymorphic DNA) primers URP4 and 7 were used. This new cultivar 'Song-A' of Pleurotus eryngii is characterized by a small number of primordia formation and the stip is thick and long. Therefore, we expect that this new strain will save of labor and cost by without culling work.

• Journal of Mushroom
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• v.17 no.4
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• pp.197-204
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• 2019

In this study, smart farm technology was used by farmers cultivating 'CHIKUMASSHU T-011' in order to develop an optimal growth model for the precision cultivation of bottle-grown winter mushroom and the results of the same are mentioned herein. Farmers participating in the experiment used 60 ㎡ of bed area with 4 rows and 13 columns of shelf shape, 20 horsepower refrigerator, 100T of sandwich panel for insulation, 6 ultrasonic humidifiers, 12 kW of heating, and 20,000 bottles of Flammulina velutipes mushroom spores. The temperature, humidity, and carbon dioxide concentrations, which directly affect the growth of the mushroom, were collected and analyzed from the environmental sensors installed at the winter mushroom cultivation area. The initial temperature was found to be 14.5℃, which was maintained at 14℃ to 15℃ until the 10th day. In the restriction phase, the initial temperature was 4℃ and was maintained between 2℃ and 3℃ until the 15th day, while during the growth phase, it was maintained between 7.5℃ to 9.5℃. Analysis of the humidity data revealed initial humidity to be 100%, which varied between 88% to 98% during primordia formation period. The humidity remained between 77% to 96% until the 15th day, in the restriction phase and between 75% to 83% during the growth phase. The initial carbon dioxide concentration was 3,500 ppm and varied between 3,500 ppm to 6,000 ppm during primordia formation period and was maintained at 6,000 ppm until the 15th day. During the growth phase, the carbon dioxide concentration was found to be over 6,000 ppm. Fruiting body characteristics of 'CHIKUMASSHU T-011' cultivated in the farmhouse were as follows: Pileus diameter of 7.5 mm and thickness of 4.1 mm, stipe thickness of 3.3 mm, and length of 154.2 mm. The number of valid fruiting bodies was 1,048 unit per 1,400 mL bottle, and the individual weight was 0.71 g per unit. The yield of fruiting bodies was 402.8 g per 1,400 mL bottle.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.342-347
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• 2004

A bacterial strain YC4963 with antifungal activity against Colletotrichum orbiculare, a causal organism of cucumber anthracnose was isolated from the rhizosphere soil of Siegesbeckia pubescens Makino in Korea. Based on physiological and biochemical characteristics and 16S ribosomal DNA sequence analysis, the bac­terial strain was identified as Pseudomonas aurantiaca. The bacteria also inhibited mycelial growth of several plant fungal pathogens such as Botrytis cinerea, Fusarium oxysporum and Rhizoctonia solani on PDA and 0.1 TSA media. The antifungal activity was found from the culture filtrate of this isolate and the active compound was quantitatively bound to XAD adsorption resin. The antibiotic compound was purified and identified as phenazine-l-carboxylic acid on the basis of combined spectral and chemical analyses data. This is the first report on the production of phenazine-l-carboxylic acid by Pseudomonas aurantiaca.

• Korean Journal of Microbiology
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• v.51 no.2
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• pp.97-107
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• 2015

Wetlands constitute a transitional zone between terrestrial and aquatic ecosystems and have unique characteristics such as frequent inundation, inflow of nutrients from terrestrial ecosystems, presence of plants adapted to grow in water, and soil that is occasionally oxygen deficient due to saturation. These characteristics and the presence of vegetation determine physical and chemical properties that affect decomposition rates of organic matter (OM). Decomposition of OM is associated with activities of various extracellular enzymes (EE) produced by bacteria and fungi. Extracellular enzymes convert macromolecules to simple compounds such as labile organic carbon (C), nitrogen (N), phosphorus (P), and sulfur (S) that can be easily taken up by microbes and plants. Therefore, the enzymatic approach is helpful to understand the decomposition rates of OM and nutrient cycling in wetland soils. This paper reviews the physical and biogeochemical factors that regulate extracellular enzyme activities (EEa) in wetland soils, including those of ${\beta}$-glucosidase, ${\beta}$-N-acetylglucosaminidase, phosphatase, arylsulfatase, and phenol oxidase that decompose organic matter and release C, N, P, and S nutrients for microbial and plant growths. Effects of pH, water table, and particle size of OM on EEa were not significantly different among sites, whereas the influence of temperature on EEa varied depending on microbial acclimation to extreme temperatures. Addition of C, N, or P affected EEa differently depending on the nutrient state, C:N ratio, limiting factors, and types of enzymes of wetland soils. Substrate quality influenced EEa more significantly than did other factors. Also, drainage of wetland and increased temperature due to global climate change can stimulate phenol oxidase activity, and anthropogenic N deposition can enhance the hydrolytic EEa; these effects increase OM decomposition rates and emissions of $CO_2$ and $CH_4$ from wetland systems. The researches on the relationship between microbial structures and EE functions, and environmental factors controlling EEa can be helpful to manipulate wetland ecosystems for treating pollutants and to monitor wetland ecosystem services.

• Journal of Applied Biological Chemistry
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• v.60 no.4
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• pp.363-372
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• 2017

Hypericum ascyron has long been used as medicinal plant and recent studies reported that H. ascyron has anti-diabetic, anti-oxidant, and anti-bacterial effects. In this study, inhibitory effect from H. ascyron on pro-inflammatory responses has been investigated. H. ascyron was extracted at optimal extraction condition. Total phenolic contents in water and 90% ethanol were 29.75 and 31.82 mg/g, respectively. Hyaluronidase inhibitory activity of H. ascyron extracts ($50-200{\mu}g/mL$ phenolics) was 0.00-14.81% and 15.33-47.49%, respectively. In cell viability, cell toxicity was shown at concentration of $100{\mu}g/mL$ and $30{\mu}g/mL$ of water and 90% ethanol extract. Therefore, $10-50{\mu}g/mL$ in water extracts and $5-20{\mu}g/mL$ in ethanol extracts was selected each for further study. Inducible nitric oxide synthase (iNOS) derived nitric oxide (NO) and cyclooxygenase (COX)-2-derived prostaglandin $E_2$ ($PGE_2$) protein expression inhibitory effect of extracts were inhibited in a dose dependent manner, significantly. Also, the pro-inflammatory cytokines inhibitory effect such as tumor necrosis $factor-{\alpha}$, nterleukin (IL)-6 and $IL-1{\beta}$ were decreased in the dose dependent manner. The results indicate that H. ascyron extracts reduced inflammatory responses in lipopolysaccharide-induced 264.7 cells via the regulation of the iNOS, COX-2, NO, $PGE_2$, and pro-inflammatory cytokines. Therefore, H. ascyron extracts have significant anti-inflammatory effect and a source as therapeutic materials.