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Impairment of Mitochondrial ATP Synthesis Induces RIPK3-dependent Necroptosis in Lung Epithelial Cells During Lung Injury by Lung Inflammation

  • Su Hwan Lee;Ju Hye Shin;Min Woo Park;Junhyung Kim;Kyung Soo Chung;Sungwon Na;Ji-Hwan Ryu;Jin Hwa Lee;Moo Suk Park;Young Sam Kim;Jong-Seok Moon
    • IMMUNE NETWORK
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    • v.22 no.2
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    • pp.18.1-18.15
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    • 2022
  • Dysfunction of mitochondrial metabolism is implicated in cellular injury and cell death. While mitochondrial dysfunction is associated with lung injury by lung inflammation, the mechanism by which the impairment of mitochondrial ATP synthesis regulates necroptosis during acute lung injury (ALI) by lung inflammation is unclear. Here, we showed that the impairment of mitochondrial ATP synthesis induces receptor interacting serine/threonine kinase 3 (RIPK3)-dependent necroptosis during lung injury by lung inflammation. We found that the impairment of mitochondrial ATP synthesis by oligomycin, an inhibitor of ATP synthase, resulted in increased lung injury and RIPK3 levels in lung tissues during lung inflammation by LPS in mice. The elevated RIPK3 and RIPK3 phosphorylation levels by oligomycin resulted in high mixed lineage kinase domain-like (MLKL) phosphorylation, the terminal molecule in necroptotic cell death pathway, in lung epithelial cells during lung inflammation. Moreover, the levels of protein in bronchoalveolar lavage fluid (BALF) were increased by the activation of necroptosis via oligomycin during lung inflammation. Furthermore, the levels of ATP5A, a catalytic subunit of the mitochondrial ATP synthase complex for ATP synthesis, were reduced in lung epithelial cells of lung tissues from patients with acute respiratory distress syndrome (ARDS), the most severe form of ALI. The levels of RIPK3, RIPK3 phosphorylation and MLKL phosphorylation were elevated in lung epithelial cells in patients with ARDS. Our results suggest that the impairment of mitochondrial ATP synthesis induces RIPK3-dependent necroptosis in lung epithelial cells during lung injury by lung inflammation.

Identification of key genes and carcinogenic pathways in hepatitis B virus-associated hepatocellular carcinoma through bioinformatics analysis

  • Sang-Hoon Kim;Shin Hwang;Gi-Won Song;Dong-Hwan Jung;Deok-Bog Moon;Jae Do Yang;Hee Chul Yu
    • Annals of Hepato-Biliary-Pancreatic Surgery
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    • v.26 no.1
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    • pp.58-68
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    • 2022
  • Backgrounds/Aims: Mechanisms for the development of hepatocellular carcinoma (HCC) in hepatitis B virus (HBV)-infected patients remain unclear. The aim of the present study was to identify genes and pathways involved in the development of HBV-associated HCC. Methods: The GSE121248 gene dataset, which included 70 HCCs and 37 adjacent liver tissues, was downloaded from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) in HCCs and adjacent liver tissues were identified. Gene ontology and Kyoto Encyclopedia of Genes and Genome pathway enrichment analyses were then performed. Results: Of 134 DEGs identified, 34 were up-regulated and 100 were down-regulated in HCCs. The 34 up-regulated DEGs were mainly involved in nuclear division, organelle fission, spindle and midbody formation, histone kinase activity, and p53 signaling pathway, whereas the 100 down-regulated DEGs were involved in steroid and hormone metabolism, collagen-coated extracellular matrix, oxidoreductase activity, and activity on paired donors, including incorporation or reduction of molecular oxygen, monooxygenase activity, and retinol metabolism. Analyses of protein-protein interaction networks with a high degree of connectivity identified significant modules containing 14 hub genes, including ANLN, ASPM, BUB1B, CCNB1, CDK1, CDKN3, ECT2, HMMR, NEK2, PBK, PRC1, RACGAP1, RRM2, and TOP2A, which were mainly associated with nuclear division, organelle fission, spindle formation, protein serine/threonine kinase activity, p53 signaling pathway, and cell cycle. Conclusions: This study identified key genes and carcinogenic pathways that play essential roles in the development of HBV-associated HCC. This may provide important information for the development of diagnostic and therapeutic targets for HCC.

The Biochemical Studies on Stored Soy-sauce (저장(貯藏)간장의 생화학적(生化學的) 연구(硏究))

  • Chang, Chi-Hyun
    • Applied Biological Chemistry
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    • v.9
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    • pp.9-27
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    • 1968
  • Studies were carried out in order to elucidate chemical components and microflora in three types of soy-sauce, 12-year aged soy-sauce prepared by improved method. 7-year aged and 20-year aged soy-sauce prepared by ordinary method. They results are summarized as follows: 1. The followings are found to be the important factors affecting the quality of soy sauce. a. Organic acids, reducing sugars and free amino acids were increased in the course of storage. b. In the aged soy-sauces under study non-volatile organic acid increased while volatile organic acid decreased and the total acidity was dependent only upon tie latter. c. It was found that suit concentration decreased during the storage. 2. The results of investigation of microflora in the stored soy-sauce are shown as follows. Soy-sauce Improved Ordinary Microbe 12-Y. 20-Y. 7-Y. Aerobic bacteria colony/1ml. 6 123 2 Halophilic lactic acid bacteria colony/1ml. 4 6 10 Osmophilic yeast colony/1ml. $828{\times}10^4$ 248 - b. In the stored soy-sauces, aerobic bacteria are incapable of growing due to drop in pH value and the influence of salt concentration. c. Halophilic lactic acid bacteria are incapable of growing due to drop in pH value, even the salt concentrations decreased during the storage. d. Osmophilic yeast are still growing in low pH value and in the decreasing salt concentration during the strage. 3. The results of amino acid analysis by paper partition chromatographic and calorimetric methods are shown as follows. a. Fourteen kinds of amino acid and thirteen amino acids were detected in the soy-sauce of 12-year aged improved soy-sauce and 7-year aged and 20-year aged ordinary one, respectively. b. The contents of aspartic acid, glutamic acid, serine, valine, leucine, lysine, histidine and methionine increased in the 20-year aged ordinary soy-sauce compared to the 7-year aged one. On the other hand those of alanine, tyrosine, phenylalanine and cystine decreased. 4. The results of sugar analysis by paper chromatography are as follows. a. In the 12-year aged improved soy-sauce, galactose, glucose, arabinose, xylose, rhamnose, maltose and an unknown were detected, and their amounts were in the above order except maltose and an unknown. b. Both in the 7 and 12-year aged ordinary soy-sauces, galactose, arabinose, xylose, glucose and rhamnose were detected and the amounts of the sugars were in the above order. c. In the non-aged ordinary soy-sauce, glucose was not detected but detected from 7-year and 20-year aged ordinary soy-sauce. 5. The results of organic acid analysis by paper chromatography were as follows. a. As volatile acids, acetic, propionic and butyric acids were detected in the 7-year aged ordinary soy-sauce. On the other hand in both the 20-year aged ordinary soy-sauce and the 12-year aged improved ones, only acetic acid was abundant while propionic and butyric acids were round in trace. It was found that propionic and butyric acids, as the unpleasant flavor components, decreased during the storage. b. In the ordinary soy-sauce, citric acid were produced during the storage and lack, malic and tartaric acids increased in the course of aging while succinic, glycolic, fumaric and malonic acids were shown to decrease. Glutaric and oxalic acids disappeared. Citric acid was produced also in the improved soy-sauce, but lactic, tartaric, succinic, malic, and glycolic acids decreased, while both malonic and glutaric acids disappeared. From the above results the citric acid production was considered to be a favorable factor for the taste. c. In the aged soy-sauces, pyruvic, α-ketoglutaric and probably acetoacetic and oxaloacetic acids (both in trace) were present and their amounts were in the above order. All of the α-keto acid abruptly decreased during the storage.

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MiR-126-3p inhibits apoptosis and promotes proliferation by targeting phosphatidylinositol 3-kinase regulatory subunit 2 in porcine ovarian granulosa cells

  • Zhou, Xiaofeng;He, Yingting;Jiang, Yao;He, Bo;Deng, Xi;Zhang, Zhe;Yuan, Xiaolong;Li, Jiaqi
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.6
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    • pp.879-887
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    • 2020
  • Objective: Numerous studies have indicated that the apoptosis and proliferation of granulosa cells (GCs) are closely related to the normal growth and development of follicles and ovaries. Previous evidence has suggested that miR-126-3p might get involved in the apoptosis and proliferation of GCs, and phosphatidylinositol 3-kinase regulatory subunit 2 (PIK3R2) gene has been predicted as one target of miR-126-3p. However, the molecular regulation of miR-126-3p on PIK3R2 and the effects of PIK3R2 on porcine GCs apoptosis and proliferation remain virtually unexplored. Methods: In this study, using porcine GCs as a cellular model, luciferase report assay, mutation and deletion were applied to verify the targeting relationship between miR-126-3p and PIK3R2. Annexin-V/PI staining and 5-ethynyl-2'-deoxyuridine assay were applied to explore the effect of PIK3R2 on GCs apoptosis and proliferation, respectively. Real-time quantitative polymerase chain reaction and Western Blot were applied to explore the regulation of miR-126-3p on PIK3R2 expression. Results: We found that miR-126-3p targeted at PIK3R2 and inhibited its mRNA and protein expression. Knockdown of PIK3R2 significantly inhibited the apoptosis and promoted the proliferation of porcine GCs, and significantly down-regulated the mRNA expression of several key genes of PI3K pathway such as insulin-like growth factor 1 receptor (IGF1R), insulin receptor (INSR), pyruvate dehydrogenase kinase 1 (PDK1), and serine/threonine kinase 1 (AKT1). Conclusion: MiR-126-3p might target and inhibit the mRNA and protein expressions of PIK3R2, thereby inhibiting GC apoptosis and promoting GC proliferation by down-regulating several key genes of the PI3K pathway, IGF1R, INSR, PDK1, and AKT1. These findings would provide great insight into further exploring the molecular regulation of miR-126-3p and PIK3R2 on the functions of GCs during the folliculogenesis in female mammals.

Comparative effect of dietary borage oil and safflower oil on anti-proliferation and ceramide metabolism in the epidermis of essential fatty acid deficient guinea pigs (필수지방산 결핍이 유도된 기니피그에서 보라지유와 홍화유 섭취의 표피 과증식 억제 및 세라마이드 대사에 미치는 효과 비교)

  • Lee, Se Ryung;Cho, Yunhi
    • Journal of Nutrition and Health
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    • v.48 no.4
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    • pp.319-326
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    • 2015
  • Purpose: Borage oil (BO) and safflower oil (SO) are efficacious in reversing epidermal hyperproliferation, which is caused by the disruption of epidermal barrier. In this study, we compared the antiproliferative effect of dietary BO and SO. Altered metabolism of ceramide (Cer), the major lipid of epidermal barrier, was further determined by measurement of epidermal levels of individual Cer, glucosylceramide (GlcCer), and sphingomyelin (SM) species, and protein expression of Cer metabolizing enzymes. Methods: Epidermal hyperproliferation was induced in guinea pigs by a hydrogenated coconut diet (HCO) for 8 weeks. Subsequently, animals were fed diets of either BO (group HCO + BO) or SO (group HCO + SO) for 2 weeks. As controls, animals were fed BO (group BO) or HCO (group HCO) diets for 10 weeks. Results: Epidermal hyperproliferation was reversed in groups HCO + BO (67.6% of group HCO) and HCO + SO (84.5% of group HCO). Epidermal levels of Cer1/2, GlcCer-A/B, and ${\beta}$-glucocerebrosidase (GCase), an enzyme of GlcCer hydrolysis for Cer generation, were higher in group HCO + BO than in group HCO, and increased to levels similar to those of group BO. In addition, epidermal levels of SM1, serine palmitoyltransferase (SPT), and acidic sphingomyelinase (aSMase), enzymes of de novo Cer synthesis and SM hydrolysis for Cer generation, but not of Cer3-7, were higher in group HCO + BO than in group HCO. Despite an increase of SPT and aSMase in group HCO + SO to levels higher than in group HCO, epidermal levels of Cer1-7, GlcCer-A/B, and GCase were similar in these two groups. Notably, acidic ceramidase, an enzyme of Cer degradation, was highly expressed in group HCO + SO. Epidermal levels of GlcCer-C/D and SM-2/3 did not differ among groups. Conclusion: Dietary BO was more prominent for reversing epidermal hyperproliferation by enhancing Cer metabolism with increased levels of Cer1/2, GlcCer-A/B, and SM1 species, and of GCase proteins.

Effect of Monosodium Glutamate and Temperature Change on the Content of Free Amino Acids in Kimchi (Monosodium Glutamate와 숙성온도 변화가 김치의 유리아미노산 함량에 미치는 영향)

  • 이예경;이명예;김순동
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.2
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    • pp.399-404
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    • 2004
  • This study was conducted to investigate the effect of monosodium glutamate (MSG) and fermentation methods (C-I; fermented for 5 days at 1$0^{\circ}C$ after 2$0^{\circ}C$ fermentation for 2 days, C-II; fermented for 7 days at 1$0^{\circ}C$, M-I; kimchi with MSG fermented for 5 days at 1$0^{\circ}C$ after 2$0^{\circ}C$ fermentation for 2 days, M-II; kimchi with MSG fermented for 7 days at 1$0^{\circ}C$ on fermentation and free amino acid content. Fermentation of M-I and M-II was slightly delayed compared to C-I and C-II. Total microbe of C-I and C-IIwere lower than those of M-I and M-II, and lactic acid bacteria of C-I and C-II were lower than those of M-I and M-II respectively. The major free amino acids were alanine, asparagine, homocystine and valine in C-I, especially, glutamic acid and ornithine were high in C-II. Homocystine, alanine, asparagine and valine in M-I, glutamic acid, alanine, hydroxyproline, asparagine, homocystine, ornithine and valine were the major free amino acid in M-II, respectively. The sour taste of M-I and M-II was lower than those of C-I and C-II, respectively, and the effect of delaying fermentation at 1$0^{\circ}C$ did not showed in the C-I and M-I. The crispy taste of the M-I and M-II was higher than those of C-I and C-II, which was the opposite results of sour taste. Palatable and overall taste of M-I and M-II were higher than those of C-I and C-II, respectively These results suggest that the MSG in kimchi affect not only increment of free amino acid content but also shelf-life and taste improvement, and continuous fermentation at 1$0^{\circ}C$ also enhance the content of free amino acid and shelf-life of kimchi.

Growth Characteristics and Variation in Amino Acids Composition of Quality Protein Maize Lines (고라이신(QPM) 주요 계통과 교잡계의 생육특성 및 아미노산 조성 변이)

  • Bae, Hwan-Hee;Son, Beom-Young;Go, Young-Sam;Park, Hye-Young;Yi, Gibum;Ha, Jun Young;Kim, Mi-Jung;Kim, Sun-Lim;Baek, Seong-Bum
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.3
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    • pp.222-230
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    • 2020
  • Maize grain quality can be improved by raising lysine content, which is an essential amino acid present in insufficient quantities in normal maize. Maize varieties with such modifications are known as quality protein maize (QPM). To date, no Korean maize cultivars contain high amounts of lysine. To introduce quality protein maize to Korean cultivars, we crossed QPM CIMMYT maize lines (CML) with KS140, an elite inbred line used as a parent of several cultivars such as 'Gangdaok' and 'Pyeonganok'. We analyzed the phenotypic characteristics of F1 plants as well as the protein contents, amino acids, and fatty acids profiles of the self-pollinated seeds of the F1 hybrids, and evaluated the feasibility of CML as a source of QPM. Days to anthesis of the CML ranged from 78 to 90 days after planting (DAP), whereas a range of 81~87 was recorded for F1 hybrids. The average days to anthesis was 85 for KS140, CML, and the F1 hybrids. The protein content of the CML was measured to be between 9.1 and 12.1%, with the highest and lowest values being recorded in CML153 and 191, respectively. The F1 hybrids had protein contents of 9.1~11.1%, and the highest content was observed in KS141/CML188. The fatty acids profiles were very similar across all analyzed maize samples, and linoleic acid (C18:2) composed the greatest proportion. Glutamic acid made up the largest proportion of amino acids in all maize samples. Lysine composition was highest in CML155 (6.92% of all amino acids), with an average composition of 4.83% across the CML. In contrast, KS140 showed a lysine content of 2.51%. In F1 hybrids, the average lysine composition was 3.46%, and KS140/CML164 (4.18%) and KS140/CML163 (4.99%) contained more lysine than either parent. Taken together, these results indicate that CML could become promising QPM sources to improve grain quality in Korean maize cultivars.

Effect of Germination on the Quality and Amino acid Composition of Soymilk (대두발아(大豆發芽)가 대두유(大豆乳)의 품질(品質) 및 아미노산(酸) 조성(組成)에 미치는 영향)

  • Kim, Woo-Jung;Oh, Hoon-Il;Oh, Myung-Won;Byun, Si-Myung
    • Korean Journal of Food Science and Technology
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    • v.15 no.1
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    • pp.12-18
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    • 1983
  • Changes in the quality and amino acid composition of soymilk prepared from soaked and germinated soybeans were investigated. Soybeans were soaked in water for 3 hrs and germinated at $18{\pm}1^{\circ}C$ for 5 days followed by water extraction at room temperature, and then the soymilk was boiled for 30 min. The initial yields of total solid and protein after soaking were 80.7% and 88.6%, respectively and decreased slowly during germination. A slow decrease in lipids and a rapid reduction in total sugar content were found during germination. The change in protein fraction content of soymilk showed an initial increase followed by a gradual decrease. The intrinsic viscosity increased rapidly after 3 days of germination to maximum value at 4th day, then decreased. The amino acid composition of protein fraction of soymilk showed little change while that of nonprotein fraction changed significantly. After 4 days of germination, aspartic acid and alanine increased more than twice, and methionine and tyrosine decreased to half of their initial composition in soymilk prepared from soaked soybean.

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Biological Activities and Cultural Characteristics of an Entomogenous fungus, Paecilomyces tenuipes (Peck) Samson (눈꽃동충하초의 배양적 특성 및 생리활성)

  • Ha, Nam-Gyu;Kim, Seung-Yul;Kang, Jin-Ho;Kang, Pil-Don;Sung, Gyoo-Byung;Hong, In-Pyo
    • Journal of Sericultural and Entomological Science
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    • v.47 no.1
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    • pp.12-17
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    • 2005
  • To develop technique for the production of P. tenuipes stromata on a large scale, the infection of P. tenuipes and the growth of stroma were investigated by silkworm (Bombyx mori) variety. Also, studied about biological activites of fruiting body formed on silkworm. Infection rate of the 5th instar larvae of the silkworm with P. tenuipes was the highest in Yangwonjam, followed by Hachojam, Baegokjam and Chilbojam in that order. Also, as the inoculation times was increased, infection rate tended to be raised. The rate of fruiting body formation of the silkworm pupae infected with P. tenuipes was the highest in Baegokjam, followed by Yangwonjam and Chilbojam in the order. But, actually the fruiting body formation of the 5th instar larvae of the silkworm tested was good in Chilbojam, followed by Yangwonjam and Baegokjam in that order in 3 times spraying inoculation. The fruiting bodies of Yangwonjam and Chilbojam infected with P. tenuipes had high amount of Mannitol, but Baegokjam and Hachojam had high concentration of Glucose on a dry weight basis. The mean content of total amino acid in the fruiting bodies of P. tenuipes was 1.03 ${\mu}mole/g$. The distribution rate of amino acid components decreased in the order of Arginine (12.2%)>Glycine (10.5%)> Proline (9.6)>Tyrosine (8.9%)>Serine>Leucine>Threonine. The most abundant amino acid in the fruiting bodies of the Baegokjam, Chilbojam and Hachojam infected with P. tenuipes was arginine, while Yangwonjam was Glycine. The most abundant fatty acid in P.tenuipes was Oleic acid on a dry weight basis. The unsaturated fatty acids such as Oleic acid, Linoleic acid and Linolenic acid accounted for more than 78% of the total fatty acids.

Studies on the Ecology of Occurrence and Identification of Typhula Snow Mold of Graminous Plants -II. Several Factors Affecting Growth of Typhula incarnata- (화본과식물에 발생하는 설부소입균핵병균(雪腐小粒菌核病菌)의 동정 및 발생상태에 관한 연구 -II. Typhula incarnata의 생육에 미치는 몇 가지 요인-)

  • Kim, Jin-Won;Lee, Du-Hyung;Shim, Gyu-Yul
    • The Korean Journal of Mycology
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    • v.20 no.1
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    • pp.37-43
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    • 1992
  • Typhula incarnata grew over a temperature range of -5 to $20^{\circ}C$ with maximum growth at 10 to $15^{\circ}C$. Sclerotial production for T. incarnata was greatest at the higher temperature. Maximum mycelial growth of this pathogen occurred from pH 5.4 to 6.2. When carbon sources were added to a basal salt medium (Czapek's dox agar) at 5 g carbon sources/l, inulin, soluble starch, galactose, glucose, mannose, manitol, sucrose, maltose, cellobirose, trehalose, raffinose, and dextrin supported growth better than other carbon sources did. Of the twenty-three nitrogen sources tested, glycine, serine, ammonium sulfate, asparagine, asparatic acid, and ${\beta}-alanine$ were the most favorable for mycelial growth of T. incarnata. Cystine and cysteine were poor nitrogen sources. Ammonium salt of nitrogen sources supported growth better than nitrate salt of nitrogen sources. Potato dextrose agar, oat meal agar, and V-8 juice agar were the most favorable for mycelial growth and sclerotial formation. Appropriate addition of pepton to PDA decreased mycelial dry weight, but sucrose supported good growth of T. incarnata. Percent viable sclerotia of T. incarnate buried in bentgrass soil decreased from 2 months after treatment remarkably. Trichoderma riride and bacteria were isolated from non-germinated sclerotia. Live orchard grass leaf pieces within the soil were colonized by T. incarnata better than sterile and unsterile dead leaf pieces at $0^{\circ}C$. Saprophytic ability of T. incarnate on sterile leaf sheath occurred better at $0^{\circ}C$ than at $10^{\circ}C$. Saprophytic microflora consisting of Cladosporium sp., Fusarium sp., Mucor sp., Pythium sp., and unidentified fungi were the competitors for the sterilized and unsterilized substrate, but their colonization was not find on live leaf sheath buried in the soil at $0^{\circ}C$. In the effect of fungicides to Typhula snow mold disease of creeping bentgrass, mixture of polyoxin and thiram was the most effective, followed by iprodione, mixture of iprodione and oxine copper, thiophanate-methyl, myclobutanil, and tolclofos-methyl.

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