• Title/Summary/Keyword: enzymatic method

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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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Diagnostic Significance of Cholesterol in Pleural Effusion (흉막액내 Cholesterol치 측정의 진단적 의의)

  • Yeo, Dong-Seung;Lee, Dong-Il;Lee, Soo-Keol;Moon, Chang-Hyung;Park, Soon-Kew;Shin, Young-Kee
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.3
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    • pp.248-254
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    • 1992
  • Background: Pleural effusion is one of the most common clinical problems in pulmonology because of high prevalence of pulmonary tuberculosis and bronchogenic carcinoma in Korea. The differential diagnosis between pleural transudate and exudate is very important, but it is very difficult in some cases. Methods: In order to assess the clinical usefulness of cholesterol levels for the differential diagnosis of pleural transudate and exudate, we measured pleural fluid cholesterol levels by enzymatic method in 45 patients who were admitted due to pleural effusion. Results: The mean cholesterol level of transudate was $33.1{\pm}12.9\;mg%$, tuberculous exudate was $97.3{\pm}28.2\;mg%$ and malignant exudate was $97.3{\pm}28.2mg%$. When the cut-off value of pleural cholesterol level was 60 mg%, one case (6.7%) of tuberculous exudate and two cases (13.3%) of malignant exudate were incorrectly classified, but all cases of transudate were classified correctly. When the cut-off value of pleural/serum cholesterol ratio was 0.3, one case (6.7%) of transudate and two cases (13.3%) of malignant exudate were incorrectly classified, but all cases of tuberculous exudate were classified correctly. When the cut-off value of pleural cholesterol level to differentiate pleural transudate from exudate was 60 mg%, sensitivity was 90% and specificity was 100%. When the cut-off value of pleural/serum cholesterol level to differentiate pleural transudate form exuidate was 0.3, sensitivity was 93% and specifiity was 93%. Conclusions: From the above results, it can be concluded that measurement of pleural fluid cholesterol levels is useful for the differential diagnosis between pleural transudate and exudate.

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Enzymatic Desugarization of Egg White for Drying with Glucose Oxidase (Glucose Oxidase에 의(依)한 건조용(乾燥用) 난백(卵白)의 효소적(酵素的) 탈당(脫糖))

  • Song, Kwang Taek;Oh, Hong Rock;Kwon, Soon Ki;Lee, Bong Duck
    • Korean Journal of Agricultural Science
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    • v.11 no.2
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    • pp.223-232
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    • 1984
  • The influences of some factors involved in removing glucose from egg white by the glucose oxidase system be fore drying were investigated. And the properties between foams prepared from raw and enzyme-treat ed egg white was compared. The results obtained we re summarized as follows; 1. The dianisidine method was found to be suitable for the measurement of egg white glucose in the range up to 100ug/ml. 2. The optimal pH of glucose oxidase activity on glucose was found to be a bout 5.0, and thats activity was most stable in the pH range of about 4.0~5.0 when that enzyme was treat ed for 30 minute at $50^{\circ}C$. 3. The optimal temperature for glucose oxidase reaction on glucose was found to be about $20^{\circ}C$, and that enzyme activity was s table up to $50^{\circ}C$. 4. The removing rate of glucose from egg white with glucose oxidase was influenced by the enzyme concentration, pH and oxygen addition, and the react ion time of the desugarization was about 10 hour sunder the conditions of 0.5% hydrogen peroxide, pH 7.0 and $26^{\circ}C$. 5. All of the each egg white treated with glucose oxidase, glucose oxidase+pancreatin, glucose oxidase+trypsin showed highly foaming ability than that of natural egg white(control), but thats foam stability, on the contrary, was reversed.

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Studies on the Fermentation on Soy Sauce by Bacteria (세균(細菌)을 이용(利用)한 간장 제조(製造)에 관(關)한 연구(硏究))

  • Ju, Hyun-Kyu;Ro, Sin-Kyu;Im, Moo-Hyun
    • Korean Journal of Food Science and Technology
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    • v.4 no.4
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    • pp.276-284
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    • 1972
  • A strain of Bacillus sp. which has powerful enzymatic activity and sets free a little amount of ammonia nitrogen and large amount of amino nitrogen was separated from Meju, Denzang, Chungkukjang, and paddy straw to make the soy-sauce rapidly and conveniently in the various mixing ratio of steamed soy-bean and wheat parched or steamed. Total nitrogen and amino nitrogen were increased during the maturing. The acidity of soy-sauce was increased to $1.15{\sim}1.98%$ than ordinary soy-sauce. pH were ranged in $4.6{\sim}6.0%$. The fermenting time takes 30 days while it takes at least 4 monthes in ordinary method. The best results were obtained with the mixed ratio of 1 : 1 or 1 : 2 (wheat : soy-bean).

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Comparative Studies on the Enzymatic Properties of Trypsins from Cat-shark and Mackerel -1. Purifications and Reaction Conditions of the Trypsins- (복상어와 고등어의 Trypsin에 관한 비교 효소학적 연구 -1. Trypsin의 정제와 반응조건-)

  • PYEUN Jae-Hyeung;CHO Deuk-Moon;HEU Min-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.24 no.5
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    • pp.273-288
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    • 1991
  • To elucidate the physiological and biochemical differences between chondrichthyes and osteichthyes, the properties of the specific digestive enzymes in cat-shark, Cephaloscyllium umbratile, and mackerel, Scomber japonicus, were studied. Homogenous trypsin proved through the disc-electrophoresis, SDS-PAG electrophoresis and gel filtration was obtained from the pancreas of cat-shark by $50-70\%$ saturated ammonium sulphate fractionation, DEAE-Sephadex A-50 column chromatography, benzamidine-Sepharose 6B affinity chromatography and Sephadex G-75-120 gel filtration. Two types of trypsins were also obtained from the pyloric caeca of mackerel by $30-70\%$ saturated ammonium sulphate fractionation and the slightly modified procedure from the method adopted in the purification of cat-shark trypsin. The two trypsins, designated trypsin A and B, were proved their homogeneity by disc- and SDS-PAG electrophoresis and gel filtration. The molecular weights of the trypsins were estimated to be 31,700 for cat-shark trypsin, 30,000 for mackerel trypsin A and 29,000 for mackerel trypsin B by SDS-PAG electrophoresis, but those were estimated to be 21,500 for cat-shark trypsin, 23,700 for mackerel trypsin A and 21,500 for mackerel trypsin B by gel filtration. The trypsins exhibited their optimum conditions at pH 9.0 and on temperature ranged from $45^{\circ}C\;to\;50^{\circ}C$ for cat-shark, and at pH 8.0 and a temperature of $50^{\circ}C$ for mackerel trypsin A and B, respectively. The cat-shark trypsin was stable at pH 10.0 and the temperature below $10^{\circ}C$, whereas the mackerel trypsin A and B, were stable in the range over pH 7.0 to pH 9.0 below $10^{\circ}C$ and at pH 8.0 below $35^{\circ}C$, respectively. The mackerel trypsins were severely inhibited by some heavy metal ions such as $Ag^{2+},\;Cu^{2+}\;and\;Hg^{2+}$ compared to cat-shark trypsin. All of the enzymes were also inhibited by antipain, leupeptin, TLCK(tosyllysine chloromethyl ketone) and SBTI(soybean trypsin inhibitor) remarkably. The inhibitory effects of PMSF(phenylmethane sulphonylfluoride), DFP(diisopropyl fluorophosphate) and benzamidine were indicated that these enzymes belong to serine-proteases.

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Effect of Environmental Factors on the Growth of Rabbit Oral Keratinocytes (토끼 구강점막 상피세포 성장에 미치는 환경인자의 영향)

  • Yoon, Moon-Young;Park, Hee-Jung;Lee, Doo-Hoon;Jang, In-Keun;Park, Jung-Keug;Kim, Woo-Sik
    • Korean Chemical Engineering Research
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    • v.43 no.1
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    • pp.103-109
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    • 2005
  • Isolation and primary culture technique of rabbit oral keratinocytes, and the study for effect of environmental factors on the cell growth were carried out in T75-flask. $1.92{\pm}0.59{\times}10^6$ viable cells were isolated by trypsin enzymatic digestion method from $0.25cm^2$ biopsy of rabbit oral mucosa. Primary culture with 10 mL of K-SFM containing 50 mg/L BPE, $5.0{\mu}g/L$ EGF and 0.15 mM $Ca^{2+}$ showed confluence after 8 days and doubling time was 2.54 days. Effect of medium types, medium volume and supplement types on the cell growth was investigated after the cultured keratinocytes had been harvested from primary confluence. Serum addition showed adverse effect and the increase of serum concentration didn't have an effect on the cell growth. The increase of medium volume decreased the cell growth. The increase of calcium concentration increased the cell growth and 2.0 mM was optimum value. In conclusion, when rabbit oral keratinocytes was cultured in T75-flask, the most effective conditions was to use 10 mL of K-SFM containing 50 mg/L BPE, $5.0{\mu}g/L$ EGF and 2.0 mM $Ca^{2+}$, and doubling time was 1.32 days. This study can provide the useful informations to develop a process and design a bioreactor for the culture of keratinocytes in human body like skin and cornea, as well as mucosa.

Some Properties of the Polyphenol Oxidase from Potatoes (Solanum tubersum L.) and Inhibiting Effect of the Polyphenol Oxidase by Sulfites (감자 Polyphenol Oxidase의 효소학적 성질 및 아황산염에 의한 활성억제 효과)

  • Ha, Young-Duk;Lee, Mi-Ock
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.17 no.3
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    • pp.198-204
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    • 1988
  • This study was aimed at obtaining elementary data on enzymatic browning of potato and potato products and examining the inhibitory method of browning. Therefore, we extracted polyphenol oxidase from potatoes(Solanum tubersum L.), and investigates its general properties and inhibiting effects of its activity with the different concentrations of sulfites($Na_2S_2O_4,\;Na_2SO_3{\cdot}7H_2O,\;NaHSO_3$). The optimum pH and temperature of polyphenol oxidase were observed to be 6.5 and $37^{\circ}C$ respectively. The polyphenol oxidase at PH5 was very stable, and the activity of polyphenol oxidase between pH $5.0{\sim}9.0$ was estimated to be relatively high, showing $72{\sim}75%$ of its activity at pH5. The polyphenol oxidase was very stable when heated at $40^{\circ}C$ for one hour, and almost 50% of enzyme activity was decreased when heated at $70^{\circ}C$ for twelve minutes. At 0.1mM concentrating of sulfites the relative activity of polyphenol oxidase was 98% in all the three cases of sulfites. Thus sulfites at 0.1mM concentration was found to have little inhibiting effect on polyphenol oxidase activity. At 1mM concentration of sulfites $NaHSO_3$ showed the lowest 36% relative activity among the three. At 5mM concentration of sulfites, the relative activity of $Na_2SO_3{\cdot}7H_2O$ was the lowest 14%.

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Studies on the Hydrolysis of the Waste wood of Cortinellus edodes with Trichoderma viride Cellulase (표고재배폐재(栽培廢材)의 당화(糖化)에 관(關)한 연구(硏究))

  • Min, Du Sik
    • Journal of Korean Society of Forest Science
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    • v.43 no.1
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    • pp.31-34
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    • 1979
  • In this study, enzymatic hydrolysis of the substrate of the waste wood of Cortinellus edodes was investigated using crude cellulase preparation of Trichoderma viride Pers. ex. Fr. SANK 16374. The crude cellulase was produced by the submerged culture process and produced in the culture fluid was salted out quantitatively by the use of ammonium sulfate. Reducing sugar was determined by the dinitrosalisylic acid (DNS) method. 1. The chemical composition of the waste wood was crude protein 2.26%, c. fat 2.57%, c. fibre 44.60%, c. ash 5.58% and lignin 13.62%. In amino acid composition, no cystine and methionine was showed, but trace amount of Vitamin A, $B_1$, and $B_2$, niacine and chloride were detected. (Table 1) 2. As heat treatment of the substrate was found to produce the highest reducing sugar yield being reacted for 48hr. with T.v cellulase, the substrate was heated to $190{\pm}5^{\circ}C$. for 45 min. either before or immediately after milling. 3. The substrate heated and ball milled at $190{\pm}5^{\circ}C$. for 45 min. the reducing sugar yield reached to 11.5%. 4. The substrate without any treatment was found to produce the highest reducing sugar yield being reacted 72hr. with T. v cellulase, the reducing sugar yield reached to 10.1%. 5. The rate of reducing sugar per each treated substrate was decreased by the order of the substrated, heated and then ball milled at $190{\pm}5^{\circ}C$. for 45 min. (11.5%)> without any treatment (10.1)> ball milled and heated at $190{\pm}5^{\circ}C$. for 45 min. (6.9%). 6. Saccharification of waste wood has been shown to be possible by heat treated and milling the substrate in contact with cellulase. And it is likely to be recommended that the waste wood may be valuable for raw materials of saccharification.

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