• Title/Summary/Keyword: Biological effects

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The Study of Cosmeceutical Activities from Lentinula edodes extracts and Application a Natural Cosmetic Material (표고버섯 추출물의 화장품약리활성 검증과 천연화장품 소재로써의 활용에 관한 연구)

  • Seo, Myeong-Seong;Jang, Young-Ah;Lee, Jin-Tae
    • Journal of the Korean Applied Science and Technology
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    • v.35 no.4
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    • pp.1003-1012
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    • 2018
  • This study is for checking the possibility of Lentinula edodes as cosmetic materials. For this we carried out biological active evaluation about anti-oxidant and anti-inflammatory effects by Lentinula edodes extracts. We extracted Lentinula edodes with water and 70% ethanol and then in order to evaluate anti-oxidant activity we treated samples by concentrations (100, 500, 1000) ${\mu}g/ml$ and carried out 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, the activity of 2,2'-azino-bis ( 3-ethylbenzothiazoline-6-sulphonic acid )-diammonium salt (ABTS) cation radical scavenging and superoxide dismutase(SOD) like activity. Also, in order to evaluate effect of anti-inflammatory the samples in macrophages(RAW 264.7 cells), we carried out evaluation of cell viability, nitric oxide inhibitory activity western blot. The results of DPPH, $ABTS^+$ radical scavenging activity and SOD-like activity of the Lentinula edodes extracts increased in dose-dependent manner. The cytotoxic of samples by MTT assay showed no toxicity at the concentrations of 10, 25 and $50{\mu}g/ml$ of Lentinula edodes extract. Nitric oxide inhibition activity results showed that the extracts reduced NO productions in a concentration-dependent manner. Expression of inflammatory cytokines as $TNF-{\alpha}$, $PGE_2$ and $IL-1{\beta}$ decreased in a concentration-dependent manner and iNOS and COX-2 proteins expression rates were decreased significantly in western blot analysis. From the results of the experiment, it was comfirmed that the Lentinula edodes extracts had excellent anti-oxidant and anti-inflammatory effect and could be used as a safe natural cosmetic material in the future.

Effect of Semisulcospira libertina Extracts from Different Extraction Processes on Liver Cell Toxicity and Ethanol Metabolism (간세포 독성과 에탄올 대사에서 추출 조건에 따른 다슬기 추출물의 효과)

  • Cho, Kyoung Hwan;Choo, Ho Jin;Seo, Min Gyun;Kim, Jong Cheol;Shin, Yu Jin;Ryu, Gi Hyung;Cho, Hee Young;Jeong, Chi-Young;Hah, Young-Sool
    • Food Engineering Progress
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    • v.21 no.2
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    • pp.158-166
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    • 2017
  • Although Semisulcospira libertina is generally regarded as a supplement for the alleviation of alcohol hangover, little is known about its effects on cell metabolism. Therefore, this study was conducted to analyze the constituents of the extracts prepared using different extraction methods and to compare their biochemical properties. The amino acid contents were found to be much higher in acidic and enzymatic hydrolysates than hot water extracts from S. libertina. DPPH radical scavenging activities in acidic and enzymatic hydrolysates were higher than those of hot water extracts. Three types of S. libertina hydrolysate was added to HepG2 cells damaged by acetaminophen (AAP), after which the survival rate of HepG2 cell were measured. In addition, lactate dehydrogenase (LDH) activities in the culture media were evaluated. The survival rates of HepG2 cells were $77.0{\pm}4.3%$ and $81.5{\pm}1.3%$ at 3 h and 5h enzymatic hydrolysates, respectively. These cell survival rates were higher compared to those of the negative control group ($67.8{\pm}4.3%$) treated only with acetaminophen. Cellular toxicities induced by treatment with AAP were also significantly alleviated in response to treatment with the extracts of S. libertina. In addition, the activities of 2 key enzymes that metabolize ethanol, alcohol dehydrogenase and aldehyde dehydrogenase, were upregulated by 4.7- and 2.7-fold respectively in response to treatment with a 3 h enzymatic hydrolysate of S. libertina. Taken together, these results provide biochemical evidence of the method by which S. libertina exerts its biological functions, including the alleviation of alcohol hangover and the protection of liver cells against toxic insults.

Optimization of cultivation conditions for pullulan production from Aureobasidium pullulans MR by response surface methodology (반응표면분석법을 이용한 Aureobasidium pullulans MR의 풀루란 생산을 위한 배양 조건 최적화)

  • Jo, Hye-Mi;Kim, Ye-Jin;Yoo, Sang-Ho;Kim, Chang-Mu;Kim, KyeWon;Park, Cheon-Seok
    • Korean Journal of Food Science and Technology
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    • v.53 no.2
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    • pp.195-203
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    • 2021
  • Aureobasidium pullulans, a black yeast, produces pullulan, a linear α-glucan composed of maltotriose repeating units linked by α(1→6)-glycosidic linkages. Pullulan can be widely used in food, cosmetic, and biotechnology industries. In this study, we isolated eight strains of A. pullulans from Forsythia koreana, Magnolia kobus DC., Spiraea prunifolia var. simpliciflora, Cornus officinalis, Cerasus, and Hippophae rhamnoides. Among them, A. pullulans MR was selected as the best pullulan producer. The effects of a carbon source, a nitrogen source, and pH on pullulan production were examined. The optimal cultivation conditions for pullulan production by A. pullulans MR were determined by response surface methodology as 15% sucrose, 0.4% soy peptone, and an initial pH of 7 at 26℃. Under these conditions, the predicted pullulan production was 47.6 g/L, which was very close to the experimental data (48.9 g/L).

Characterization of Antidiabetic Compounds from Extract of Torreya nucifera (비자나무 추출물의 항당뇨 활성물질의 특성 연구)

  • Kim, Ji Won;Kim, Dong-Seob;Lee, Hwasin;Park, Bobae;Yu, Sun-Nyoung;Hwang, You-Lim;Kim, Sang Hun;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.32 no.1
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    • pp.1-10
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    • 2022
  • Natural products have gained increasing attention due to their advantage of long-term safety and low toxicity for a very long time. Torreya nucifera is widespread in southern Korea and Jeju Island and its seeds are commonly used as edible food. Oriental ingredients have often been reported for their insecticidal, antioxidant and antibacterial properties, but there have not yet been any studies on their antidiabetic effect. In this study, we investigated several biological activities of T. nucifera pericarp (TNP) and seeds (TNS) extracts and proceeded to characterize the antidiabetic compounds of TNS. The initial results suggested that TNS extract at 15 and 10 ㎍/ml concentration has inhibitory effects on α-glucosidase and protein tyrosine phosphatase 1B, that is 14.5 and 4.35 times higher than TNP, respectively. Thus, the stronger antidiabetic TNS was selected for the subsequent experiments to characterize its active compounds. Ultrafiltration was used to determine the apparent molecular weight of the active compounds, showing 300 kDa or more. Finally the mixture was then partially purified using Diaion HP-20 column chromatography by eluting with 50~100% methanol. Therefore we concluded that the active compounds of TNS have potential as therapeutic agents in functional food or supplemental treatment to improve diabetic diseases.

Comparison of Anti-inflammatory, Skin Barrier Improvement, and Anti-aging Efficacy of Eleutherococcus divaricatus var. chiisanensis and various Eleutherococcus Genus Extract (지리산오갈피, 가시오갈피, 오갈피나무, 오가나무 추출물의 항염증, 피부장벽개선, 항노화 효능 비교)

  • Jiwon, Han;Bomi, Nam;Beom seok, Lee;Jin-A, Ko;Jiyoung, Hwang
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.4
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    • pp.373-383
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    • 2022
  • Inflammation caused by active oxygen and the resulting barrier damage have been consistently pointed out as the cause of wrinkle formation. In this study, effective index ingredient search and efficacy analysis were performed to verify the value of use as a functional cosmetic material related to antioxidant, anti-inflammatory and skin barrier improvement, and anti-aging for extracts of four types of Eleutherococcus divaricatus var. chiisanensis (ED), Eleutherococcus senticosus (EN), Eleutherococcus sessiliflorus (ES), and Eleutherococcus sieboldianus (EI) belonging to the Eleutherococcus genus. To identify the effective index composition, the content of the ingredients was measured by high-performance liquid chromatography. The content of eleutheroside E and chlorogenic acid was the highest in ED among the Eleutherococcus genus. As for anti-oxidant activity, DPPH radical scavenging activity was the highest in ED. In anti-inflammatory effects, ED extracts inhibited nitric oxide generation in inflammatory macrophage cells due to lipopolysaccharide by 40% at 100 ㎍/mL. In the case of IL-6 inhibition, which is known as a pro-inflammatory cytokine, ED showed 41% inhibition at 100 ㎍/mL. In addition, filaggrin and involucrin, which are skin barrier-related factors, were increased by 2.5 times and 1.6 times, respectively, in 100 ㎍/mL of ED extracts, and as for the collagenase, which is a wrinkle-related factor, ED extract showed 29% efficacy at 100 ㎍/mL. Thus, these result suggested that ED extract, among the four Eleutherococcus genus, can be used as a cosmetic ingredient for suppressing inflammation in the skin, reinforcing the skin barrier, and reducing wrinkles.

Newly Synthesized Phosphodiesterase 4 (PDE4) Inhibitor, DWP205505, Inhibits TNF-$\alpha$ Secretion and mRNA Expression

  • Lee, Suk-Kyeong;Lee, Sun-A;Byun, Hye-Sin;Cho, Mi-La;Kim, Wan-Uk;Park, Sung-Hwan;Cho, Chul-Soo;Joo, Young-Shil;Lee, Shin-Seok;Yoo, Eun-Sook;Son, Ho-Jung;Kim, Ho-Youn
    • Journal of Microbiology and Biotechnology
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    • v.9 no.1
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    • pp.106-112
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    • 1999
  • The therapeutic potential of phosphodiesterase 4(PDE4) inhibitors in inflammatory diseases including some autoimmune diseases has been explored recently with some hopeful results. These PDE4 inhibitors are thought to show their anti-inflammatory effect by down-regulating tumor necrosis factor-a (TNF-$\alpha$) production in lymphocytes and macrophages. A high concentration of TNF-$\alpha$has been found in rheumatoid arthritis (RA) synovium and reducing TNF-$\alpha$using biological agents was proven to be an effective RA treatment. To test the possibility of using PDE4 inhibitors for RA treatment, the effects of a newly synthesized PDE4 inhibitor, DWP205505, on TNF-$\alpha$ and IL-10 production was tested in cells isolated from normal peripheral blood and rheumatoid arthritis synovial fluid. Cytokine production was assayed at the protein level by sandwich enzyme-linked immunosorbent assay (ELISA) and at the mRNA expression level by semi-quantitative RT-PCR. Another PDE4 inhibitor, RP73401, was used for comparison. DWP205505 and RP73401 had no harmful effect on cell viability up to 10 $\mu$M concentration during the 24 h culture period. DWP205505 as well as RP73401 significantly reduced TNF-$\alpha$ secretion from lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (pBMC) and synovial fluid mononuclear cells (SFMC). The effect of DWP205505 or RP73401 treatment on the mRNA expression of TNF-$\alpha$ was also studied in LPS-stimulated PBMC and SFMC. TNF-$\alpha$ mRNA expression was increased by LPS stimulation and both of the PDE4 inhibitors suppressed TNF-$\alpha$ mRNA expression. For interleukin-l0 (IL-l0), a little different results were obtained from PBMC and SFMC; IL-l0 secretion was unaffected by LPS stimulation and only minimally affected by both of the PDE4 inhibitors in PBMC. In unstimulated SFMC, DWP205505 and RP73401 slightly enhanced IL-10 secretion, while they reduced IL-l0 secretion from LPS-stimulated SFMC where IL-l0 secretion was a lot higher than unstimulated SFMC. These results suggest that the newly synthesized PDE4 inhibitor DWP205505 may have anti-rheumatoid arthritis activity.

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Relationships between Above-ground Carbon Stock and Species and Structural Diversity in South Korean Forests (우리나라 산림의 지상부 탄소저장량과 종 다양성 및 구조적 다양성과의 관계)

  • Heon Mo Jeong;Eui Joo Kim;Jae Hoon Park;Hae Ran Kim
    • Korean Journal of Ecology and Environment
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    • v.57 no.3
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    • pp.123-133
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    • 2024
  • To analyse the relationship between above-ground carbon stocks, species diversity and broadleaved forests structural diversity of South Korean forests, we collected vegetation inventories from environmental impact assessment projects over the past 10 years. The available data were selected and organised including tree species, DBH and area each projects. The data was classified by forest type, aboveground carbon stocks were calculated and compared, and the correlation between aboveground carbon stocks and biodiversity and structural diversity was analysed. The results showed that above-ground carbon stocks were higher in mixed forests and broadleaved forests and lower in needleleaved forests, similar to previous studies. However aboveground carbon stocks of mixed forests were higher in natural forests than in plantations. Aboveground carbon stocks in broadleaved forests were higher in plantations than natural forests, and there was no statistical different of between natural and plantations in needleleaved forest. This could be the result of a variety influences including biological and environmental factors in the study area, and further research is needed to analyse the effects on carbon sequestration. Correlation analysis showed no correlation between biodiversity and above-ground carbon stocks, but a positive correlation between structural diversity and above-ground carbon stocks. This indicates that above-ground carbon stocks in forests are associated with unevenness diameters and the proportion and evenness of tree species by diameter. In addition, it has been analysed that the high succession stages in forest have higher species diversity and structural diversity, and greater efficiency in the utilization of resources required for plant growth, leading to increased plant productivity and storage. Considering that the study sites were young forests with an average DBH of 14.8~23.7 cm, it is expected that carbon stocks will increase as biodiversity and structural diversity increase. Further research is needed to develop techniques to quantitatively assess the relationship of diversity to carbon stocks for policy use in assessing and increasing carbon stocks in forests.

Studies on Takju Brewing with Potatoes (감자를 이용(利用)한 탁주제조(濁酒製造)에 관(關)한 연구(硏究))

  • Kim, S.Y.;Oh, M.J.;Kim, C.J.
    • Applied Biological Chemistry
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    • v.17 no.2
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    • pp.81-92
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    • 1974
  • In order to prepare the mashing materials for 'Takju', Korean wine, with potatoes, theywere steamed, dryed, and pulverized, and their chemical components were analyzed. As a brewing method of Takju with potatoes, general 2nd stage process with Ipkuk and Bunkuk (enzyme sources), commonly used now, was carried out and the effects of preparing conditions of Ipkuk(koji) with potato flour, mashing materials and brewing conditions on the contents of Takju mash, and of storing time on the contents of Takju, were investigated and the results obtained were summarized as follows, 1. Chemical components of steamed potatoes and potato flour were Moisture; 76.2, 10.8%, Total sugar; 16.1, 69.8%, Reducing sugar; 3.45, 13.4%, Crude protein; 2.1, 11.3%, Total acid; 0.012, 0.023% and Volatile acid; 0.0012, 0.0025% respectively. 2. The most effective preparing conditions of Ipkuk with potato flour were to incubate the potato flour added 40-50% of water for 48 hours by general preparing process of Koji, and liquefying and saccharogenic amylase activities of Ipkuk incubated at above conditions were $D40^{\circ}$ 30' 128 W.V. and 13.2 A.U.. 3. The effects of various brewing conditions on the contents of Takju mashes were as follows; 1) Optimum ratio of mashing water and materials for Takju brewing with potato flour was 140ml of water to 60g of flour in 1st stage and 260ml to 140g in 2nd stage. 2) Optimum fermentating times and temperatures for Takju brewing were at $25^{\circ}C$ for 48 hours in 1st stage and at $30^{\circ}C$ for 48 hours in 2nd stage. 3) Optimum amounts of enzyme sources for Takju brewing were 20-30% of Ipkuk and 0.5% of Bunkuk in 1st stage and 1.0% of Bunkuk in 2nd stage. 4) Methanol content of the Takju mash brewed with raw potato flour was much more than that with steamed potato flour. 5) Alcohol, fusel oil and Formol nitrogen contents of the Takju mash brewed with potato flour were less than that with wheat flour, on the contrary, methanol contents and total acidifies of them were showed conversely above. 4. The changes of chemical components and microflora in the mashes during the brewing potato flour Takju were as follows; 1) The accumulation of ethanol followed rapidly in early stage, being the highest at 72 hours (11.9%). 2) Total sugar content of the mash was decreased considerably within 48-72 hours, being 2.62% at 72 hours, and thereafter slowly. 3) Reducing sugar of the mash had a tendency of decreasing, being 0.29% at 48 hours. 4) Total acidity, volatile acidity and Formol nitrogen content of the mash were increased slowly, being 7.30, 0.20, 2.55 at 48 hours. 5) Total cells of yeast appeared the highest in 72 hours ($2.1{\times}10^8$) and thereafter decreased slowly. 6) Total cells of bacteria appeared the highest in 48 hours ($2.4{\times}10^8$) and thereafter decreased or increased slightly. 5. Takju was made from the fermented mash mixed with water to be 6% of alcohol content, and the change of alcohol content, total acidity, total cells of yeast and bacteria during the storing at $30^{\circ}C$ were as follows; 1) Alcohol content of Takju was increased slightly at 24 hours (6.2%), and thereafter decreased slowly. 2) Total acidity of Takju was increased gradually, being 6.1 at 72 hours 3) Total cells of yeast and bacteria appeared the highest at 48 hours ($2.3{\times}10^8,\;1.5{\times}10^8$), and thereafter decreased slowly. 6. Alcohol content, total acidity and Formol nitrogen content of the Takju brewed with potato flour Ipkuk or wheat flour Ipkuk and steamed potatoes(1:5) were 9.8-11.3%, 5.8-7.4, 2.5-3.3 respectively, and the color of the Takju. was similar to commercial Takju. 7. The results of sensory test for various experimental Takju, showed that the Takjues brewed with the materials combined with wheat flour and steamed potatoes(4:5 or 3.5:7.5) were not significantly different in color, taste and flavor from commercial Takju, However, those with potato flour and wheat flour (1:1 or 7:3) were significantly different from commercial Takju.

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Pharmacological Studies of Cefoperazone(T-1551) (Cefoperazone(T-1551)의 약리학적 연구)

  • Lim J.K.;Hong S.A.;Park C.W.;Kim M.S.;Suh Y.H.;Shin S.G.;Kim Y.S.;Kim H.W.;Lee J.S.;Chang K.C.;Lee S.K.;Chang K.C.;Kim I.S.
    • The Korean Journal of Pharmacology
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    • v.16 no.2 s.27
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    • pp.55-70
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    • 1980
  • The pharmacological and microbiological studies of Cefoperazone (T-1551, Toyama Chemical Co., Japan) were conducted in vitro and in vivo. The studies included stability and physicochemical characteristics, antimicrobial activity, animal and human pharmacokinetics, animal pharmacodynamics and safety evaluation of Cefoperazone sodium for injection. 1) Stability and physicochemical characteristics. Sodium salt of cefoperazone for injection had a general appearance of white crystalline powder which contained 0.5% water, and of which melting point was $187.2^{\circ}C$. The pH's of 10% and 25% aqueous solutions were 5.03 ana 5.16 at $25^{\circ}C$. The preparations of cefoperazone did not contain any pyrogenic substances and did not liberate histamine in cats. The drug was highly compatible with common infusion solutions including 5% Dextrose solution and no significant potency decrease was observed in 5 hours after mixing. Powdered cefoperazone sodium contained in hermetically sealed and ligt-shielded container was highly stable at $4^circ}C{\sim}37^{\circ}C$ for 12 weeks. When stored at $4^{\circ}C$ the potency was retained almost completely for up to one year. 2) Antimicrobial activity against clinical isolates. Among the 230 clinical isolates included, Salmonella typhi was the most susceptible to cefoperazone, with 100% inhibition at MIC of ${\leq}0.5{\mu}g/ml$. Cefoperazone was also highly active against Streptococcus pyogenes(group A), Kletsiella pneumoniae, Staphylococcus aureus and Shigella flexneri, with 100% inhibition at $16{\mu}g/ml$ or less. More than 80% of Escherichia coli, Enterobacter aerogenes and Salmonella paratyphi was inhibited at ${\leq}16{\mu}/ml$, while Enterobacter cloaceae, Serratia marcescens and Pseudomonas aerogenosa were somewhat less sensitive to cefoperagone, with inhibitions of 60%, 55% and 35% respectively at the same MIC. 3) Animal pharmacokinetics Serum concentration, organ distritution and excretion of cefoperazone in rats were observed after single intramuscular injections at doses of 20 mg/kg and 50 mg/kg. The extent of protein binding to human plasma protein was also measured in vitro br equilibrium dialysis method. The mean Peak serum concentrations of $7.4{\mu}g/ml$ and $16.4{\mu}/ml$ were obtained at 30 min. after administration of cefoperazone at doses of 20 mg/kg and 50 mg/kg respectively. The tissue concentrations of cefoperazone measured at 30 and 60 min. were highest in kidney. And the concentrations of the drug in kidney, liver and small intestine were much higher than in blood. Urinary and fecal excretion over 24 hours after injetcion ranged form 12.5% to 15.0% in urine and from 19.6% to 25.0% in feces, indicating that the gastrointestinal system is more important than renal system for the excretion of cefoperazone. The extent of binding to human plasma protein measured by equilibrium dialysis was $76.3%{\sim}76.9%$, which was somewhat lower than the others utilizing centrifugal ultrafiltration method. 4) Animal pharmacodynamics Central nervous system : Effects of cefoperazone on the spontaneous movement and general behavioral patterns of rats, the pentobarbital sleeping time in mice and the body temperature in rabbits were observed. Single intraperitoneal injections at doses of $500{\sim}2,000mg/kg$ in rats did not affect the spontaneous movement ana the general behavioral patterns of the animal. Doses of $125{\sim}500mg/kg$ of cefoperazone injected intraperitonealy in mice neither increased nor decreased the pentobarbital-induced sleeping time. In rabbits the normal body temperature was maintained following the single intravenous injections of $125{\sim}2,000mg/kg$ dose. Respiratory and circulatory system: Respiration rate, blood pressure, heart rate and ECG of anesthetized rabbits were monitored for 3 hours following single intravenous injections of cefoperazone at doses of $125{\sim}2,000mg/kg$. The respiration rate decreased by $3{\sim}l7%$ at all the doses of cefoperazone administered. Blood pressure did not show any changes but slight decrease from 130/113 to 125/107 by the highest dose(2,000 mg/kg) injected in this experiment. The dosages of 1,000 and 2,000 mg/kg seemed to slightly decrease the heart rate, but it was not significantly different from the normal control. All the doses of cefoperazone injected were not associated with any abnormal changes in ECG findings throughout the monitering period. Autonomic nervous system and smooth muscle: Effects of cefoperazone on the automatic movement of rabbit isolated small intestine, large intestine, stomach and uterus were observed in vitro. The autonomic movement and tonus of intestinal smooth muscle increased at dose of $40{\mu}g/ml$ in small intestine and at 0.4 mg/ml in large intestine. However, in stomach and uterine smooth muscle the autonomic movement was slightly increased by the much higher doses of 5-10 mg/ml. Blood: In vitro osmotic fragility of rabbit RBC suspension was not affected by cefoperazone of $1{\sim}10mg/ml$. Doses of 7.5 and 10 mg/ml were associated with 11.8% and 15.3% prolongation of whole blood coagulation time. Liver and kidney function: When measured at 3 hours after single intravenous injections of cefoperaonze in rabbits, the values of serum GOT, GPT, Bilirubin, TTT, BUN and creatine were not significantly different from the normal control. 5) Safety evaluation Acute toxicity: The acute toxicity of cefoperazone was studied following intraperitoneal and intravenous injections to mice(A strain, 4 week old) and rats(Sprague-Dawler, 6 week old). The LD_(50)'s of intraperitonealy injected cefoperazone were 9.7g/kg in male mice, 9.6g/kg in female mice and over 15g/kg in both male and female rats. And when administered intravenously in rats, LD_(50)'s were 5.1g/kg in male and 5.0g/kg in female. Administrations of the high doses of the drug were associated with slight inhibition of spontaneous movement and convulsion. Atdominal transudate and intestinal hyperemia were observed in animals administered intraperitonealy. In rats receiving high doses of the drug intravenously rhinorrhea and pulmonary congestion and edema were also observed. Renal proximal tubular epithelial degeneration was found in animals dosing in high concentrations of cefoperazone. Subacute toxicity: Rats(Sprague-Dawley, 6 week old) dosing 0.5, 1.0 and 2.0 g/kg/day of cefoperazone intraperitonealy were observed for one month and sacrificed at 24 hours after the last dose. In animals with a high dose, slight inhibition of spontaneous movement was observed during the experimental period. Soft stool or diarrhea appeared at first or second week of the administration in rats receiving 2.0g/kg. Daily food consumption and weekly weight gain were similar to control during the administration. Urinalysis, blood chemistry and hematology after one month administration were not different from control either. Cecal enlargement, which is an expected effect of broad spectrum antibiotic altering the normal intestinal microbial flora, was observed. Intestinal or peritoneal congestion and peritonitis were found. These findings seemed to be attributed to the local irritation following prolonged intraperitoneal injections of hypertonic and acidic cefoperazone solution. Among the histopathologic findings renal proximal tubular epithelial degeneration was characteristic in rats receiving 1 and 2g/kg/day, which were 10 and 20 times higher than the maximal clinical dose (100 mg/kg) of the drug. 6) Human pharmacokinetics Serum concentrations and urinary excretion were determined following a single intravenous injection of 1g cefoperazone in eight healthy, male volunteers. Mean serum concentrations of 89.3, 61.3, 26.6, 12.3, 2.3, and $1.8{\mu}g/ml$ occured at 1,2,4,6,8 and 12 hours after injection respectively, and the biological half-life was 108 minutes. Urinary excretion over 24 hours after injection was up to 43.5% of administered dose.

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Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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