• Title/Summary/Keyword: chemical binding

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Thermal Stability of Israeli Carp Actomyosin and Its Protection by Chemical Additives (이스라엘 잉어 Actomyosin의 열안정성과 그 보호)

  • NAM Taek-Jeong;CHOI Yeung-Joon;PYEUN Jae-Hyeung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.17 no.4
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    • pp.271-279
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    • 1984
  • Effects of temperature and additives on the stability of actomyosin extracted from skeletal muscle of Israeli carp, Cyprinus carpio nudus, were studied by analyzing free SH-group, ATP-sensitivity and Ca-ATPase activity. The used additives were sucrose, sorbitol, Na-glutamate and L-cysteine. Furthermore, the denaturation constant($K_D$), protective effect(${\Delta}E/M$) and the other thermo-dynamic parameters on protein denaturation are systematically discussed. The actomyosin showed $4.12{\sim}4.68 mg/ml$ in protein concentration, $2.63{\sim}2.93\%$ in ribonucleic acid to the protein, $1:2.20{\sim}2.63$ in the binding ratio of myosin and actin, $4.33{\sim}5.26\%$ in fat content, 109.78 in ATP-sonsitivity, $0.159{\sim}0.201\;{\mu}M-Pi/min/mg-protein$ in Ca-ATPase activity and $3.3{\sim}3.4M/10^5$g-protein in free SH-group content. The first-order rate plots were obtained on the decrease of Ca-ATPase activity and ATP-sensitivity with an increase in temperature, while the free SH-group was increased to $60^{\circ}C$ and decreased rapidly above the temperature. The half-life of Ca-ATPase activity on the actomyosin Ca-ATPase was 280 min at $12^{\circ}C$, 125 min at $20^{\circ}C$, 55 min at $30^{\circ}C$ and 13 min at $40^{\circ}C$, and activation energy, activation enthalpy, activation entropy and free energy of the proteins at $20^{\circ}C$ wene 5,395 cal/mole, 4,814 cal/mole, -40.42 e.u. and 17,626 cal/mole, respectively. The protective effect of the additives on the actomyosin Ca-ATPase showed that the most effective material is $3\%$ sorbitol and followed in the order of $8\%$ Na-glutamate, $1\%$ sucrose and $1\%$ L-cysteine. The actomyosin was more stable at $-30^{\circ}C$ than at $0^{\circ}C$ and $-20^{\circ}C$. and when the additives were used in the low temperature storage, $8\%$ Na-glutamate was the most effective. $3\%$ sorbitol, $1\%$ sucrose and $1\%$ L-cysteine was to become lower in the order.

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The Role of Organic Amendments with Different Biodegradability in Ammonia Volatilization during Composting of Cattle Manure (우분뇨 퇴비화 중 암모니아 휘산에 대한 이분해성 및 난분해성 유기 첨가물의 역할)

  • Lim, Sang-Sun;Park, Hyun-Jung;Lee, Sun-Il;Lee, Dong-Suk;Kwak, Jin-Hyeob;Choi, Woo-Jung
    • Korean Journal of Environmental Agriculture
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    • v.28 no.1
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    • pp.20-24
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    • 2009
  • This study was conducted to investigate the roles of co-existed organic materials (OM) with different biodegradability in composting of cattle manure in terms of $CO_2$ emission and $NH_3$ volatilization. Either sawdust (SD, low biodegradability) or rice bran (RB, high biodegradability) was mixed with cattle manure at a various rate and the amounts of $CO_2$ emission and $NH_3$ volatilization were determined periodically during 4 weeks of composting. Percentage of dry matter loss during the composting period was also calculated. The amount of $CO_2$ emitted increased with increasing rate of OM and was significantly (P<0.01) higher in the RB treatment than in the SD treatment by 43 to 122% depending on the rate of OM Accordingly, % of dry matter loss during 4 weeks of composting was higher in the RB (rang: from 35.1 % to 41.5%) than that in the SD treatments (from 18.7% to 22.6%), showing that RB is more biodegradable than SD. During the early composting period up to 8 days, negligible amount of ammonia volatilization was detected in both treatments regardless of application rates. In the RB treatment, substantial amount of ammonia volatilization was detected thereafter, however, no meaningful ammonia volatilization was observed in the SD treatment until the end of composting. Such differences could be attributed to the different properties of SD and RB. For example, the high C/N ratio of SD could enhance $NH_4^+$ immobilization and thus decrease $NH_4^+$ concentration that is susceptible to ammonia volatilization. Binding of $NH_4^+$ on to phenolic compounds of SD may also contribute to the decrease in $NH_4^+$ concentration. Meanwhile, as RB has a relatively low C/N ratio, remineralization of immobilized $NH_4^+$ could increase $NH_4^+$ concentration as high as the level for the occurrence of ammonia volatilization. Therefore, our study suggests that OM which is resistant to biodegradation can reduce $NH_3$ volatilization largely by physico-chemical pathways across the entire composting period and that easily biodegradable OM can retard $NH_3$ volatilzation via microbial immobilization in the early period of composting followed by rapid remineralization, leading to substantial volatilization of $NH_3$ in the middle stage of composting.

Basic Studies for the Breeding of High Protein Rice. I. Comparison of the analytical methods for the measurement of the protein content in the brown rice (수두 고단백 계통육성을 위한 기초적 연구 I. 계통육성을 위한 조단백질 분석법의 비교)

  • Mun-Hue Heu;Hak-Soo SUH
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.12
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    • pp.1-5
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    • 1972
  • In order to compare the analytical efficiency of the Kjeldahal, Dye binding and Biurett method for the determination of nitrogen content in the brown rice, correlation coefficients were calculate with the analytical data obtained by the above mentioned 3 different methods for the brown rice of 36 varieties or lines grown at 5 different nitrogen levels (0, 7.5, 15.0, 22.5 and 30.0kg/10a). Analysis of variance were made for the data of 6 varieties among those 36, and compared the precision of the data obtained by the 3 analytical methods. The expenditure (in terms of chemicals and labour) required for the 3 methods are also compared. The results are summarized as follows; 1. The correlation between D. B. C. and Kjeldahl value were generally more significant than the correlation between the value of Biurett and the value of Kjeldahl. But, the D. B. C. method generally overestimates than the Kjeldahl method at both extreme low and extreme high nitrogen contents, and the Biurett method includes more dispersed error than other two methods, though the optical values are parallel to the Kjeldahl nitrogen values at any levels of nitrogen applied. 2. The varietal difference in nitrogen value obtained by the 3 methods were different at the different nitrogen level applied. That is the interaction between variety and analytical method, and between the nitrogen level and analytical method were significant statistically. 3. The coefficient of variance (C, V.) was largest in the data analyzed by Biurett method and next in the data analayred by D. B. C. method. In the data analyzed by Biurett, the C. V. increased along onglong increase of nitrogen applied. But, in the data obtained by D. B. C. or Biurett the C. V. increased along the decrease of nitrogen applied. 4. From the comparison of the expenditure (in terms of chemicals and labour) required for the analysis of 100 samples by 3 methods, it was noticed that, the Biurett or D. B. C. method largely curtail the chemical expenditure and labour costs. Especially the Biurett method could curtail more labour costs.

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Effect of the Preparation Method on the Activity of CeO2-promoted Co3O4 Catalysts for N2O Decomposition (촉매 제조방법에 따른 Co-CeO2 촉매의 N2O 분해 특성 연구)

  • Kim, Hye Jeong;Kim, Min-Jae;Lee, Seung-Jae;Ryu, In-Soo;Yi, Kwang Bok;Jeon, Sang Goo
    • Clean Technology
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    • v.24 no.3
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    • pp.198-205
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    • 2018
  • This study investigated the influence of catalyst preparation on the activity of $Co-CeO_2$ catalyst for $N_2O$ decomposition. $Co-CeO_2$ catalysts were synthesized by co-precipitation and incipient wetness impregnation. In order to estimate the performance of the as prepared catalysts, direct catalytic $N_2O$ decomposition test was carried out under $250{\sim}375^{\circ}C$. As a result, the catalyst prepared by co-precipitation (CoCe-CP) showed an enhanced performance on $N_2O$ decomposition reaction even in the presence of $O_2$ and/or $H_2O$, whereas the impregnation catalyst (CoCe-IM) did not. In order to investigate the difference in catalytic activity, characterization such as XRD, BET, TEM, $H_2-TPR$, $O_2-TPD$, and XPS was conducted. It is confirmed that the particle size and specific surface area were changed depending on the catalyst preparation method and the synthesis process influenced the physical properties of the catalysts. In addition, the improvement in the activity of the catalyst prepared by co-precipitation is due to the enhanced reduction from $Co^{3+}$ to $Co^{2+}$ and the improved oxygen desorption rate. However, it has been confirmed that the surface electron state and binding energy, which are related to $N_2O$ decomposition, do not change depending on the preparation method.

EFFECT OF BENZALKONIUM CHLORIDE ON DENTIN BONDING WITH NTG-GMA/BPDM AND DSDM SYSTEM (Benzalkonium Chloride가 NTG-GMA/BPDM계 및 DSDM계 상아질접착제의 접착성능에 미치는 영향)

  • Shin, Il;Park, Jin-Hoon
    • Restorative Dentistry and Endodontics
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    • v.20 no.2
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    • pp.699-720
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    • 1995
  • This study was conducted to evaluate the effect of benzalkonium chloride solution as a wetting agent instead of water on dentin bonding with NTG-GMA/BPDM system (All-bond 2, Bisco.) and DSDM system (Aelitebond, Bisco.). Benzalkonium chloride solution is a chemical disinfectant widely used in medical and dental clinics for preoperative preparation of skin and mucosa due to its strong effect of cationic surface active detergent. Eighty freshly extracted bovine lower incisor were grinded labially to expose flat dentin surface, and then were acid-etched with 10 % phosphoric acid for 15 second, water-rinsed, and dried for 10 second with air syringe. The specimens were randomly divided into 8 groups of 10 teeth. The specimens of control group were remoistured with water and the specimens of experimental groups were remoistured with 0.1 %, 0.5 %, and 1.0 % benzalkonium chloride solution respectively. And then, the Aelitefil composite resin was bonded to the pretreated surface of the specimens by use of All-bond 2 dentin bonding system or Aelitebond dentin bonding system in equal number of the specimens. The bonded specimens were stored in $37^{\circ}C$ distilled water for 24 hours, then the tensile bond strength was measured, the mode of failure was observed, the fractured dentin surface were examined under scanning electron microscopy, and FT-IR spectroscopy was taken for the purpose of investigating the changes of the dentin surface pretreated with benzal konium chloride solution followed by each primer of the dentin bonding systems. The results were as follows : In the group of bonding with NTG-GMA/BPDM dentin bonding agent(All-bond 2), higher tensile bond strength was only seen in the experimental group remoistured with 0.1 % benzal konium chloride solution than that in water-remoistured control group(p<0.05). In the group of bonding with DSDM dentin bonding agent (Aelitebond), no significant differences were seen between the control and each one of the experimental group(p<0.05). Higher tensile bond strength were seen in NTG-GMAIBPDM dentin bonding agent group than in DSDM dentin bonding agent group regardless of remoistur ization with benzal konium chloride solution. On the examination of failure mode, cohesive and mixed failure were predominantly seen in the group of bonding with NTG-GMAIBPDM dentin bonding agent, while adhesive failure was predominantly seen in the group of bonding with DSDM dentin bonding agent. On SEM examination of fractured surfaces, no differences of findings of primed dentin surface between the groups with and without remoisturization with benzal konium chloride solution. FT-IR spectroscopy taken from the control and the experimental group reve::.led that some higher absorbance derived from the primers binding to dentin surface was seen at the group pretreated with 0.1 % benzal konium chloride solution than at the control group of remoisturizing with water.

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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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The Effects of Isopropyl 2-(1,3-dithioetane-2-ylidene)-2-[N-(4-methyl-thiazol-2-yl)carbamoyl]acetate (YH439) on Potentiated Carbon Tetrachloride Hepatotoxicity (상승적 화학적 간독성에 미치는 YH439의 영향)

  • Kim, Sang-Geon;Cho, Joo-Youn
    • The Korean Journal of Pharmacology
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    • v.32 no.3
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    • pp.407-416
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    • 1996
  • The reactive intermediates formed during the metabolism of therapeutic agents, toxicants and carcinogens by cytochromes P450 are frequently capable of covalently binding to tissue macromolecules and causing tissue damage. It has been shown that YH439, a congener of malotilate, is effective in suppressing hepatic P450 2E1 expression. The present study was designed to further establish the mechanistic basis of YH439 protection against toxicant by assessing its effects against chemical-mediated potentiated hepatotoxicity. Retinoyl palmitate (Vit-A) pretreatment of rats for 7 days substantially enhanced carbon tetrachloride hepatotoxicity, as supported by an ${\sim}5-fold$ increase in serum alanine aminotransferase (ALT) activity, as compared to $CCl_4$ treatment alone. The elevation of ALT activity due to Vit-A was completely blocked by the treatment of $GdCl_3$ a selective inhibitor of Kupffer cell activity. Concomitant pretreatment of rats with both YH439 and Vit-A resulted in a 94% decrease in Vit-A-potentiated $CCl_4$ hepatotoxicity. YH439 was also effective against propyl sulfide-potentiated $CCl_4-induced$ hepatotoxicity. Whereas propyl sulfide (50 mg/kg, 7d) enhanced $CCl_4-induced$ hepatotoxicity by >5-fold, relative to $CCl_4$ treatment alone, concomitant treatment of animals with both propyl sulfide and YH439 at the doses of 100 and 200 mg/kg prevented propyl sulfide-potentiated $CCl_4$ hepatotoxicity by 35% and 90%, respectively. Allyl sulfide, a suppressant of hepatic P450 2E1 expression, completely blocked the propyl sulfide-enhanced hepatotoxicity, indicating that propyl sulfide potentiation of $CCl_4$ hepatotoxicity was highly associated with the expression of P450 2E1 and that YH439 blocked the propyl sulfide-enhanced hepatotoxicity through modulation of P450 2E1 levels. Propyl sulfide- and $CCl_4-induced$ stimulation of lipid peroxidation was also suppressed by YH439 in a dose-related manner, as supported by decreases in malonedialdehyde production. The role of P450 2E1 induction in the potentiation of $CCl_4$ toxicity and the effects of YH439 were further evaluated using pyridine as a P450 2E1 inducer. Pyridine pretreatment substantially enhanced the $CCl_4$ hepatotoicity by 23-fold, relative to $CCl_4$ alone. YH439, however, failed to reduce the pyridine-potentiated toxicity, suggesting that the other form(s) of cytochroms P450 inducible by pyridine, but not suppressible by YH439 treatment, may play a role in potentiating $CCl_4-induced$ hepatotoxicity. YH439 was capable of blocking cadmium chloride-induced liver toxicity in mice. These results demonstrated that YH439 efficiently blocks Vit-A-enhanced hepatotoxiciy through Kupffer cell inactivation and that the suppression of P450 2E1 expression by YH439 is highly associated with blocking of propyl sulfide-mediated hepatotoxicity.

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Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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Identification of Sorption Characteristics of Cesium for the Improved Coal Mine Drainage Treated Sludge (CMDS) by the Addition of Na and S (석탄광산배수처리슬러지에 Na와 S를 첨가하여 개량한 흡착제의 세슘 흡착 특성 규명)

  • Soyoung Jeon;Danu Kim;Jeonghyeon Byeon;Daehyun Shin;Minjune Yang;Minhee Lee
    • Economic and Environmental Geology
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    • v.56 no.2
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    • pp.125-138
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    • 2023
  • Most of previous cesium (Cs) sorbents have limitations on the treatment in the large-scale water system having low Cs concentration and high ion strength. In this study, the new Cs sorbent that is eco-friendly and has a high Cs removal efficiency was developed by improving the coal mine drainage treated sludge (hereafter 'CMDS') with the addition of Na and S. The sludge produced through the treatment process for the mine drainage originating from the abandoned coal mine was used as the primary material for developing the new Cs sorbent because of its high Ca and Fe contents. The CMDS was improved by adding Na and S during the heat treatment process (hereafter 'Na-S-CMDS' for the developed sorbent in this study). Laboratory experiments and the sorption model studies were performed to evaluate the Cs sorption capacity and to understand the Cs sorption mechanisms of the Na-S-CMDS. The physicochemical and mineralogical properties of the Na-S-CMDS were also investigated through various analyses, such as XRF, XRD, SEM/EDS, XPS, etc. From results of batch sorption experiments, the Na-S-CMDS showed the fast sorption rate (in equilibrium within few hours) and the very high Cs removal efficiency (> 90.0%) even at the low Cs concentration in solution (< 0.5 mg/L). The experimental results were well fitted to the Langmuir isotherm model, suggesting the mostly monolayer coverage sorption of the Cs on the Na-S-CMDS. The Cs sorption kinetic model studies supported that the Cs sorption tendency of the Na-S-CMDS was similar to the pseudo-second-order model curve and more complicated chemical sorption process could occur rather than the simple physical adsorption. Results of XRF and XRD analyses for the Na-S-CMDS after the Cs sorption showed that the Na content clearly decreased in the Na-S-CMDS and the erdite (NaFeS2·2(H2O)) was disappeared, suggesting that the active ion exchange between Na+ and Cs+ occurred on the Na-S-CMDS during the Cs sorption process. From results of the XPS analysis, the strong interaction between Cs and S in Na-S-CMDS was investigated and the high Cs sorption capacity was resulted from the binding between Cs and S (or S-complex). Results from this study supported that the Na-S-CMDS has an outstanding potential to remove the Cs from radioactive contaminated water systems such as seawater and groundwater, which have high ion strength but low Cs concentration.