• Title/Summary/Keyword: Enzymatic method

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Studies on the Hydrolysis of Holocellulose with Trichoderma viride Cellulase. (III). Effects of the Optimum Treated Conditions and Reactivation of Residue of Digested Substrates (Cellulase에 의한 목재당화(木材糖化)에 관(関)한 연구(硏究) - (III) 최적(最適) 처리조건(処理條件)과 효소처리(酵素処理) 잔사(殘渣)의 재기질화(再基質化) 효과(效果) -)

  • Min, Du Sik
    • Journal of Korean Society of Forest Science
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    • v.49 no.1
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    • pp.1-5
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    • 1980
  • In this study, enzymatic saccharification of substrates from Alnus hirsuta Ruper (8-14 years). Quercus acutissima Carruthers, Betula platyphylla var. japonica Nera, Populus euramericana Guiner and Platanus orientalis L. were investigated using crude cellulase preparations of Trichoderma viride Pers. ex. Fr. SANK 16374, and conduced on the optimum treated conditions of the cellulase sacchrification and reactivation of residue of digested substrates. The Trichoderma viride cellulase was produced by the submerged culture process and produced in the culture fluid was salted out quantitatively by the use of ammonium sulfate. The method of dilignification from wood (5 species) was treated by the peracetic acid(PA) method. The reducing sugar was determined by the dinitrosalicylic acid (DNS) method. 1. The results of tests carried out for 96 hr. (Figure 1), show conclusively the initial substrates from 5 species ($S_3$) which has been rendered highly reactive form and the mean rate of reducing sugar was 28.3 %. 2. The results of tests carried out for 96 hr., the reactivation of residue of digested substrates (improvement in the quality of the substrate through preheating in air at $190^{\circ}C$. for 45 min. followed by milling was (60 mesh size) at the same substrate level, increased concentrations of cellulase at the same substrate level, and increased concentrations of cellulase increases the rate of hydrolysis considerably. 3. Figure 1. shows conclusively that the residue of digested substrates ($S_1$ dried at $60^{\circ}C$) which has been rendered extremly resistant to cellulase action can be reactivated into a highly reactive form ($S_2$), almost comparable to that of the initial substrates ($S_3$). And the reducing sugar formation did not show statistically significent differences at 5% levels by initial substrates and the residue of digested substrates (preheating in air at $190^{\circ}C$. for 45 min. fallowed by milling was (60 mesh size).

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Direct PCR Detection of the Causal Agents, Soybean Bacterial Pustule, Xanthomonas axonopodis pv. glycines in Soybean Seeds (콩 종자에서 Xanthomonas axonopodis pv. glycines의 검출을 위한 Direct PCR 방법 개발)

  • Lee, Yong-Ju;Kang, Mi-Hyung;Noh, Tae-Hwan;Lee, Du-Ku;Lee, Geon-Hwi;Kim, Si-Ju
    • Research in Plant Disease
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    • v.15 no.2
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    • pp.83-87
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    • 2009
  • Direct Polymerase Chain Reaction (PCR) method that combines biological and enzymatic amplification of PCR targets was developed for the detection of Xanthomonas axonopodis pv. glycines on soybeen seeds without DNA isolation. Primers Xag F1 and Xag R1 were designed to specifically amplify a 401 bp fragment of the glycinecin A gene of X axonopodis pv. glycines. Xag F1 and Xag R1 were used to carry out the PCR analysis with genomic DNA from 45 different bacterial strains including phylogenetically related bacteria with X axonopodis pv. glycines, and other bacterial strains of different genus and species. The PCR assay using this set of primers were able to detect X axonopodis pv. glycines with DNA concentration as low as 200 fg and $1.8{\times}10^3$ cfu/ml. The Xag was detected from the seed samples incubated for 2 hrs with shaking and the intensity of the band was increase with the incubation time of seeds. The Direct PCR assay method without DNA isolation makes detection of X. axonopodis pv. glycines on soybean seeds easier and more sensitive than other conventional methods. The developed seed assay using direct PCR method will be useful for the specific detection of X. axonopodis pv. glycines in soybean seed samples.

Method Validation for the Determination of Eleutherosides and β-Glucan in Acanthopanax koreanum (탐라오가피의 Eleutheroside B, E 및 β-Glucan 함량 분석 및 분석법 검증)

  • Kim, Young-Hyun;Bae, Da-Bin;Park, Sun-Ok;Lee, Sang-Jong;Cho, Ok-Hyun;Lee, Ok-Hwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.9
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    • pp.1419-1425
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    • 2013
  • The aim of this study was to investigate the method validation for the determination of eleutherosides (B and E) and ${\beta}$-glucan in Acanthopanax (A.) koreanum. This medicinal plant reportedly mainly included eleutherosides which exhibit the pharmacological effects, and ${\beta}$-glucan substantially enhances the function of the immune system by activating macrophages. The specificity, linearity, precision, accuracy, limit of detection (LOD, S/N=3), and limit of quantification (LOQ, S/N=10) were measured by HPLC and enzymatic methods. Our results showed that the coefficient of calibration correlation ($R^2$) for eleutheroside B and E were 0.9997 and 0.9999, respectively. The limits of detection (LOD) for eleutheroside B and E were $0.050{\mu}g/mL$ and $0.025{\mu}g/mL$, respectively. The recovery rate of eleutheroside B and E were revealed in the high range of 100.66~110.04% and 94.26~111.62%, respectively. The inter-day precision of eleutheroside B and E in the root and stem in A. koreanum were 1.4~5.0% and 1.1~2.5%, respectively. The intra-day precision of eleutheroside B and E in the root and stem in A. koreanum were 2.8~2.9% and 0.4~1.1%, respectively. Furthermore, the inter-day and intra-day precision of ${\beta}$-glucan in the stem, leaf, and fruit of A. koreanum were 1.32~5.67% and 8.01~11.76%, respectively. In conclusion, the methods were validated for the detection of eleutherosides and ${\beta}$-glucan in A. koreanum.

Improvement of an Simultaneous Determination for Clenbuterol and Ractopamine in Livestock Products using LC-MS/MS (LC-MS/MS를 이용한 축산물 중 clenbuterol과 ractopamine의 동시 분석법 개선)

  • Cho, Yoon-Jae;Chae, Young-Sik;Kim, Jae-Young;Kim, Hyung-Soo;Kang, Ilhyun;Do, Jung-Ah;Oh, Jae-Ho;Kwon, Kisung;Chang, Moon-Ik
    • Korean Journal of Food Science and Technology
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    • v.45 no.1
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    • pp.25-33
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    • 2013
  • Clenbuterol and ractopamine, which are ${\beta}$-agonists, have been misused as a growth promoting agent in meat producing animals. Clenbuterol was banned for veterinary drug in Korea because of its problems regarding safety. Due to their adverse effects, such as cardiovascular and central nervous diseases on human health proper control and monitoring should be conducted. The existing analytical method of clenbuterol and ractopamine in the Food code was improved through our present study. The bovine muscle samples were subjected to enzymatic hydrolysis, extracted with ethyl acetate and defatted by hexane-methanol partitioning. A molecular imprinted polymer (MIP) solid phase extraction cartridge was used for clean-up and LC-MS/MS was operated in positive multiple reaction monitoring (MRM). Clenbuterol-$d_9$ and ractopamine-$d_3$ were used as an internal standard. The renewed method was validated according to the CODEX guideline. The limits of quantitation for clenbuterol and ractopamine were 0.2 and 0.5 ${\mu}g/kg$, respectively. The mean recoveries ranged in 104.2-113.5% for clenbuterol and in 107.6-118.1% for ractopamine. The improved method was able to save both time and expenses.

Effects of Intracavitary Urokinase Instillation in Complicated Pleural Effusion (합병성 흉막 삼출에 대한 국소적 Urokinase 주입치료 효과에 관한 연구)

  • Sohn, Dong-Hyun;Yoon, Su-Mi;Kim, Chung-Mi;Park, Ik-Soo;Sohn, Jang-Won;Yang, Seok-Chul;Yoon, Ho-Joo;Shin, Dong-Ho;Park, Sung-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.3
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    • pp.357-364
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    • 2000
  • Background : Complicated exudative pleural fluid collections have traditionally been treated by either closed tube thoracostomy drainage or by open surgical drainage. Complete drainage is important in order to control pleural sepsis, restore pulmonary function, and entrapment. Recently intracavitary fibrinolytic therapy has been advocated as a method to facillitate drainage of complicated exudative pleural effusion and to allow enzymatic debridemant of the restrictive fibrinous sheets covering the pleural surface. The purpose of this study is to prospectively evaluate the effects of image-guided catheter drainage with high dose urokinase(UK) instillation in the treatment of complicated pleural effusions. Patients : Twenty complicated pleural effusion patients that poorly respond to image-guided drainage were allocated to receive UK. There were 8 pneumonia and 12 tuberculosis. Methods : Drugs were diluted in 250 mL normal saline and were infused intrapleurally through the chest tube or pig-tail catheter in a daily dose of 250,000 IU of UK. Response was assessed by clinical outcome, fluid drainage, chest radiography, pleural ultrasound and/or computed tomography. Results : The mean UK instillation time was $1.63{\pm}0.10$. The mean volume drained UK instillation was $381.3{\pm}314.4\;mL$, and post-UK was $321.6{\pm}489.5\;mL$. The follow up duration after UK therapy was mean $212.9{\pm}194.5$ days. We had successful results in 19 cases (95.0%). There were 12 pleural thickenings (60.0%), 2 markedly decreased effusions (10.0%) and 5 cases of no thickening or effusion. There was recurrence after treatment in only one patient(5%) with complicated pleural effusiondue to tuberculosis. Conclusions : Image-guided drainage with high dose UK instillation (250,000 U/day) in complicated pleural effusion is a safe and more effective method than closed thoracostomy drainage. And this management, in turn, can obviate surgery in most cases.

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The Predictors of Effectiveness on Urokinase Instillation Therapy into Loculated Pleural Effusion. (소방이 형성된 흉막질환에서 유로키나제 주입치료의 예후인자)

  • Song, Kee-San;Bang, Jei-So;Kwak, Seung-Min;Cho, Chul-Ho;Park, Chan-Sup
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.621-628
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    • 1997
  • Background : As the pleural inflammation progresses, exudative pleural fluid becomes loculated rapidly with pleural thickening. Complete drainage is important to prevent pleural fibrosis, entrapment and depression of lung function. Intrapleural urokinase instillation therapy has been advocated as a method to facilitate drainage of gelatinous pleural fluid and to allow enzymatic debriment of pleural surface. This study was designed to investigate the predictors of effectiveness of intrapleural urokinase in the treatment of loculated pleural effusion. Method : Thirty-five patients received a single radiographically guided pig-tail catheter ranging in size from 10 to 12 French. Twenty-two patients had tuberculous pleural effusions, and 13 had non-tuberculous postpneumonic empyemas. A total of 240,000 units of urokinase was dissolved in 240 ml of normal saline and the aliquots of 80mL was instilled into the pleural cavity via pig-tail catheter per every 8hr. Effectiveness of intrapleural urokinase instillation therapy was assessed by biochemical markers, ultrasonography, and technical details. A greater than 50% improvement on follow-up chest radiographs was defined as success group. Result : Twenty-seven of 35 (77.1%) patients had successful outcome to urokinase instillation therapy. Duration of symptoms before admission was shorter in success group ($11.8{\pm}6.9day$) than in failure group ($26.62{\pm}16.5day$) (P<0.05). Amount of drained fluid during urokinase therapy was larger in success group ($917.1{\pm}392.7ml$) than in failure group ($613.8{\pm}259.7ml$) (P<0.05). Pleural fluid glucose was higher in success group ($89.7{\pm}35.9mg/dl$) than in failure group ($41.2{\pm}47.1mg/dl$) (P<0.05). Pleural fluid LDH was lower in success group ($878.4{\pm}654.3IU/L$) than in failure group ($2711.1{\pm}973.1IU/L$) (P<0.05). Honeycomb septated pattern on chest ultrasonography was observed in six of eight failure group, but none of success group (P<0.05). Conclusion : Longer duration of symptoms before admission, smaller amount of drained fluid during urokinase therapy, lower glucose value, higher LDH value in pleural fluid examination, and honeycomb septation pattern on chest ultrasonograph were predictors for failure group of intrapleural urokinase instillation therapy.

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Dehydration of Lactic Acid to Bio-acrylic Acid over NaY Zeolites: Effect of Calcium Promotion and KOH Treatment (NaY 제올라이트 촉매 상에서 젖산 탈수반응을 통한 바이오아크릴산 생산: Ca 함침 및 KOH 처리 영향)

  • Jichan, Kim;Sumin, Seo;Jungho, Jae
    • Clean Technology
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    • v.28 no.4
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    • pp.269-277
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    • 2022
  • With the recent development of the biological enzymatic reaction industry, lactic acid (LA) can be mass-produced from biomass sources. In particular, a catalytic process that converts LA into acrylic acid (AA) is receiving much attention because AA is used widely in the petrochemical industry as a monomer for superabsorbent polymers (SAP) and as an adhesive for displays. In the LA conversion process, NaY zeolites have been previously shown to be a high-activity catalyst, which improves AA selectivity and long-term stability. However, NaY zeolites suffer from fast deactivation due to severe coking. Therefore, the aim of this study is to modify the acid-base properties of the NaY zeolite to address this shortcoming. First, base promoters, Ca ions, were introduced to the NaY zeolites to tune their acidity and basicity via ion exchange (IE) and incipient wetness impregnation (IWI). The IWI method showed superior catalyst selectivity and stability compared to the IE method, maintaining a high AA yield of approximately 40% during the 16 h reaction. Based on the NH3- and CO2-TPD results, the calcium salts that impregnated into the NaY zeolites were proposed to exit as an oxide form mainly at the exterior surface of NaY and act as additional base sites to promote the dehydration of LA to AA. The NaY zeolites were further treated with KOH before calcium impregnation to reduce the total acidity and improve the dispersion of calcium through the mesopores formed by KOH-induced desilication. However, this KOH treatment did not lead to enhanced AA selectivity. Finally, calcium loading was increased from 1wt% to 5wt% to maximize the amount of base sites. The increased basicity improved the AA selectivity substantially to 65% at 100% conversion while maintaining high activity during a 24 h reaction. Our results suggest that controlling the basicity of the catalyst is key to obtaining high AA selectivity and high catalyst stability.

Some Tertiary Interactions in 5S rRNA from Xanthomonas celebensis (Xanthomonas celebensis 5S rRNA의 몇 가지 삼차상호작용)

  • Bongrae Cho;Yeonghoon Lee;Myung-Un Choi;Inwon Park
    • Journal of the Korean Chemical Society
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    • v.37 no.2
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    • pp.237-243
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    • 1993
  • The primary and secondary structure of the 5S rRNA isolated from Xanthomonas celebensis were determined by enzymatic and chemical degradation methods. It consists of 119 nucleotides and contains no modified nucleosides. As with the 5S rRNAs of X. maltophilia and X. citri, it contains an additional uridine residue on the 5'-terminus. Its secondary structure was almost identical to the models previously proposed by us for the 5S rRNA of two Xanthomonas species. Its secondary structure consists of five helices, five loops and two bulges. The tertiary interactions in the 5S rRNA molecule were analyzed by Fe(II)-EDTA treatment and hybridization method using deoxyhexamer. From the fact that some adenine residues in loop M, region $I_1-C$, loop $H_1$, and loop $H_2$ become susceptible to diethylpyrocarbonate when the 5S rRNA was hybridized with deoxyhexamer complementary to the sequence $U_{35}CCCAU_{40}$ and that some nucleotide residues in loop M, loop $H_1$ and region $D-I_2$ become resistant Fe(II)-EDTA cleavage in the presence of $Mg^{2+}$, it is presumed that loops $H_1$ and $H_2$ interact with loop M in some way. In the tertiary interaction, the regions $I_1-C$ and $D-I_2$ seem to act as hinges in folding the stems $B-I_1-C$ and $D-I_2-E.$ It was found that loop $H_1$ changes into a smaller loop of three bases by forming noncanonical A : C base-pairs ih acidic environment.

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Autometallography for Zinc Detection in the Central Nervous System (중추신경계통내 분포하는 Zinc의 조직화학적 동정)

  • Jo, Seung-Mook;Gorm, Danscher;Kim, Sung-Jun;Park, Seung-Kook;Kang, Tae-Cheon;Won, Moo-Ho
    • Applied Microscopy
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    • v.30 no.4
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    • pp.347-355
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    • 2000
  • Zinc is one of the most abundant oligoelements in the living cell. It appears tightly bound to some metalloproteins and nucleic acids, loosely bound to some metallothioneins or even as free ion. Small amounts of zinc ions (in the nanomolar range) regulate a plentitude of enzymatic proteins, receptors and transcription factors, thus rolls need accurate homeostasis of zinc ions. Zinc is an essential catalytic or structural element of many proteins, and a signaling messenger that is released by neural activity at many central excitatory synapses. Growing evidences suggest that zinc may also be a key mediator and modulator of the neuronal death associated with transient global ischemia and sustained seizures, as well as perhaps other neurological disease stoles. Some neurons have developed mechanisms to accumulate zinc in specific membrane compartment ('vesicular zinc') which can be evidenced using histochemical techniques. Substances giving a bright colour or emitting fluorescence when in contact with divalent metal ions are currently used to detect them inside cells; their use leads to the so called 'direct' methods. The fixation and precipitation of metal ions as insoluble salt precipitates, their maintenance along the histological process and, finally, their demonstration after autometallographic development are essential steps for other methods, the so called 'indirect methods'. This study is a short report on the autometallograhical approaches for zinc detection in the central nervous system (CNS) by means of a modified selenium method.

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Clinical Experiences of Continuous Tepid Blood Cardioplegia; Valvular Heart Surgery (미온혈 심정지액의 임상적 고찰)

  • 이종국;박승일;조재민;원준호;박묘식
    • Journal of Chest Surgery
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    • v.32 no.2
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    • pp.130-137
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    • 1999
  • Background: In cardiac surgery, hypothermia is associated with a number of major disadvantage, including its detrimental effects on enzymatic function, energy generation and cellular integrity. Warm cardioplegia with normothermic cardiopulmonary bypass cause three times more incidence of permanent neurologic deficits than the cold crystalloid cardioplegia with hypothermic cardiopulmonary bypass. Interruptions or inadequate distribution of warm cardioplegia may induce anaerobic metabolism and warm ischemic injury. To avoid these problems, tepid blood cardioplegia was recently introduced. Material and Method: To evaluate whether continuous tepid blood cardioplegia is beneficial in clinical practice during valvular surgery, we studied two groups of patients matched by numbers and clinical characteristics. Warm group(37$^{\circ}C$) consisted of 18 patients who underwent valvular surgery with continuous warm blood cardioplegia. Tepid group(32$^{\circ}C$) consisted of 17 patients who underwent valvular surgery with continuous tepid blood cardioplegia. Result: Heartbeat in 100% of the patients receiving continuous warm blood cardioplegia and 88.2% of the patients receiving continuous tepid blood cardioplegia converted to normal sinus rhythm spontaneously after removal of the aortic cross clamp. There were no differences between these two groups in CPB time, ACC time, the amount of crystalloid cardioplegia used and peak level of potassium. During the operation, the total amount of urine output was more in the warm group than the tepid group(2372${\pm}$243 ml versus 1535${\pm}$130 ml, p<0.01). There were no differences between the two groups in troponin T level measured 1hr and 12hrs after the operation. Conclusion: Continuous tepid blood cardioplegia is as safe and effective as continuous warm blood cardioplegia undergoing cardiac valve surgery in myocardial protection.

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