• Title/Summary/Keyword: Enzymatic

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Uptake of the Fresh and Aged Residues of Carbofuran by Rice Plants from Soil (침투성(浸透性) 살충제(殺蟲劑) Carbofuran 토양(土壤) 잔류물(殘留物)의 벼에 의(依)한 흡수(吸收))

  • Lee, Jae-Koo;Kyung, Kee-Sung;Wheeler, W.B.
    • Korean Journal of Environmental Agriculture
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    • v.8 no.2
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    • pp.103-118
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    • 1989
  • In order to investigate the uptake of the systemic insecticide, carbofuran, 2,3-dihydro-2,2-dimethyl-7-benzofuranyl-N-methyl(arbamate) residues, fresh and aged, by rice plants, they were grown for 42 days in soils containing freshly treated (T-1), 3-month-aged (T-2), and 6-month-aged residues (T-3). The amounts of $^{14}CO_2$ evolved from $^{14}C-carbofuran$ during the 3-and 6-month aging in soil (temp. $22{\pm}1^{\circ}C$ ; moisture, 50% of the maximum water-holding capacity) were 8.9 and 26.7% of the original radioactivity applied, respectively. Mineralization of $^{14}C-carbofuran$ in soil to $^{14}CO_2$ during 42 days of rice growing was 4.4% (T-1), 11.0% (T-2), and 15.7 (T-3). The methanol extract of the 3-and 6-month-aged soils revealed that 3-keto carbofuran phenol (2,3-dihydro-2,2-dimethyl-3-oxo-7-benzofuranol) was the major metabolite, where as 3-hydroxy carbofuran (2,3-dihydro-2,2-dimethyl-3-hydroxy-7-benzofuranyl-N-methylcarbamate) turned out to be the major metabolite in the shoots by the enzymatic cleavage of the possible conjugate present in the methanol extract. Volatilization of $^{14}C-carbofuran$ in soil during 3-and 6-month-aging, and 42 days of rice growing was 0.026, 0.05, and 0.012-0.018% of the applied radioactivity, respectively. The $^{14}C-radioactivity$ which was absorbed from the soils by rice plants during 42 days of the growing period and persisted in rice plant tissues was 26.8, 21.4, and 10.3% in T-1, T-2, and T-3, respectively. The non-extractable bound residues were 8.3, 37.9, and 54.6% of the originally applied carbofuran in T-1. T-2, and T-3, respectively. The small translocation of $^{14}C-radioactivity$ in T-3 upwards suggests that major metabolite 3-keto carbofuran phenol is conjugated in roots and the low recovery in T-1 indicates the loss of carbofuran from the shoots.

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Single Cell Dissociation Methods for Flow Cytometric Cell Death Analysis of Hypoxia-Ischemia Injured Newborn Rat Pup Brain (저산소성 허혈성 뇌손상이 유발된 신생백서에서 단일세포의 분리)

  • Hwang, Jong Hee;Sung, Dong Kyung;Choi, Chang Won;Kang, Saem;Chang, Yun Sil;Park, Won Soon;Lee, Munhyang
    • Clinical and Experimental Pediatrics
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    • v.48 no.5
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    • pp.545-550
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    • 2005
  • Purpose : Newborn brain tissue has to be dissociated into a single cell suspension for flow cytometric analysis of cell death during hypoxia-ischemia. Thus the development of a method to dissociate cells from the brain tissue with least damage and maintenance of membrane and antigen integrity remains the challenge for the in vivo application of this technique. We evaluated the efficacy of mechanical or enzymatic (collagenase or tryspin) methods of brain tissue disaggregation. Methods : The extent of the damage to the plasma membrane and loss of the characteristics of the membrane induced with each dissociation method was determined by comparing the flow cytometric results labeled with both fluorescent annexin V and propidium iodide of the newborn rat pup brain tissue in the control group (n=10) and in the 48-hour after hypoxia-ischemia group (n=10). Results : In the control group, the cell percentage of damaged, apoptotic and necrotic cells of both hemispheres with the mechanical dissociation method was significantly increased compared to the trypsin or collagenase method. In the 48-hour after hypoxia-ischemia group, the cell percentage of apoptotic and necrotic cells of the right hemisphere with the collagenase method significantly increased, and live cells significantly decreased compared to the left hemisphere, control group. Although the same trend was observed, the extent of alterations made with the trypsin method was significantly less compared to the collagenase method. Conclusion : The dissociation of neonatal brain tissue for flow cytometric analysis with collagenase was most efficacious with the least cell damage and preservation of the plasma membrane characteristics.

On-site Output Survey and Feed Value Evaluation on Agro- industrial By-products (농산업부산물들에 대한 배출 현장 조사 및 사료적 가치 평가)

  • Kwak, W. S.;Yoon, J. S.
    • Journal of Animal Science and Technology
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    • v.45 no.2
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    • pp.251-264
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    • 2003
  • This study was conducted to make on-site survey on the output pattern and utilization situation of 19 by-products selected, to evaluate their nutritional characteristics, to find out a reliable index with which digestion of by-products can be predicted on the basis of chemical compositions analyzed and to diagnose the risk of using book values in the absence of the actual values analyzed for diet formulation. Production and utilization situations of by-products were quite various. Nutritionally, fruit processing by-products such as apple pomace (AP), pear pomace (PP), grape pomace (GP), and persimmon peel (PSP), and bakery by-products (BB) were classified as energy feeds. Soybean curd meal (SCM), animal by- products such as blood (BD), feather meal (FM) and poultry by-products (PB), and activated milk processing sludge (AMS) were classified as protein feeds. Soy hulls (SH), spent mushroom compost (SMC), barley malt hulls (BMH), waste paper (WP) and broiler litter (BL) were classified as roughage. Rumen contents (RC) and restaurant food waste (FW) were nutritionally analogous to complete diets for cattle and swine, respectively. Compared to soybean meal (SBM), BD and FM contained high (P<0.05) levels of amino acids and barley malt sprouts (BMS), AMS and FW contained low (P<0.05) levels of amino acids. Enzymatic (pepsin) digestibilities of proteinaceous feeds ranged between 99 and 66%. In vitro DM digestibility was high (P<0.05) in the order of FW, BB, AP, SH, PP, PSP, BMH, BMS, SCM, GP, RC, PB, BL, WP, SMC, AMS, FM and BD. In vitro DM digestibility had the highest correlation (r=0.68) with nonfibrous carbohydrate among chemical components. Differences between analyzed values of chemical components and book values were considerable. Caution is required in using book values when large amount of by-products are used in diets.

Enzymatic Hydrolysis of Yellowfin Sole Skin Gelatin in a Continuous Hollow Fiber Membrane Reactor (연속식 중공사막 반응기를 이용한 각시가자미피 젤라틴의 가수분해)

  • KIM Se-Kwon;BYUN Hee-Guk;KANG Tae-Jung;SONG Dae-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.26 no.2
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    • pp.120-132
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    • 1993
  • A continuous hollow fiber membrane reactor(CHFMR) was developed and optimized for the production of yellowfin sole(Limanda aspera) skin gelatin hydrolysates using trypsin. The results were summerized as follows: The $K_m$ value of the CHFMR was 2.4 times higher than that of the batch reactor, indicating reduced enzyme affinity for the substrate. The $K_2$ value of the CHFMR was 8.5 times lower than that of the batch process, showing a significant reduction in trypsin activity in the CHFMR. The optimum operating conditions for the CHFMR process were $55^{\circ}C$, pH 9.0, flux 7.79 ml/min, residence time 77min, and trypsin to substrate ratio, 0.01(w/w) After operating for 60min under the above conditions, $79\%$ of the total amount of initial gelatin was hydrolysed. Enzyme leakage was observed through the 10,000 MWCO membrane after the 20min of reactor operation, while none occurred after 5hr. Total enzyme leakage was about $12.95\%$ at $55^{\circ}C$ for 5hrs. However, there was no apparent correlation between enzyme leakage and substrate hydrolysis. The membrane has a significant effect on trypsin activity loss for 60min of the CHFMR operation. The CHFMR operating with the membrane lost $34\%$ of the initial activity versus a $23\%$ loss of activity after 3hr in the continuous reactor lacking the hollow fiber membrane. The measurement of fouling property showed that relative flux reduction was $91\%$ and flux recover rate was $92\%$ at $10\%$ substrate solution. The productivity(378.85mg product/mg enzyme) of the CHFMR was more than 4 times higher than that of the batch reactor at $55^{\circ}C$.

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Purification and Enzymatic Characteristics of the Bacillus pasteurii Urease Expressed in Escherichia coli (Escherichia coli에서 발현된 Recombinant Bacillus pasteurii Urease의 정제 및 효소학적 특성)

  • 이은탁;김상달
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.519-526
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    • 1992
  • The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

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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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Shalf Life Enhancement of Minimally Processed Fruits and Vegetables

  • Kim, Dong-Man
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 1993.12a
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    • pp.6-9
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    • 1993
  • According to changes in population, economic conditions, life-stile and eating habits, the frui ts and vegetables market wi 11 be shi fted from processed (i. e. , canned) to fresh. Undressed fresh produce, consisting of washed, disinfected and peeled fruits and vegetables that either sliced or grated, are currently increased in demand by retail and institutional market which use them as salad components or in ready-to use foods, Main attributes of minimally processed fruits and vegetables are convenience and fresh-like quality. Minimally processed Products readily deteriorate in quality, especially color and texture, as a result of endogeneous enzyme enhanced respiration and microorganisms which lead to reduced shelf Iife. According to changes in population, economic conditions, life-stile and eating habits, the frui ts and vegetables market wi 11 be shi fted from processed (i. e. , canned) to fresh. Undressed fresh produce, consisting of washed, disinfected and peeled fruits and vegetables that either sliced or grated, are currently increased in demand by retail and institutional market which use them as salad components or in ready-to use foods, Main attributes of minimally processed fruits and vegetables are convenience and fresh-like quality. Minimally processed Products readily deteriorate in quality, especially color and texture, as a result of endogeneous enzyme enhanced respiration and microorganisms which lead to reduced shelf Iife. Thus. to prevent these undesirable changes , val'ious techniques such as controlled atmosphere (CA) storage, modified atmosphere OIA) storage, including vacuum packaging have been receiving considerable attention, Although milch research has been done to find optimal conditions for whole intact frui ts and vegetables, only limi ted information is avai lable on fresh cut. and other minimally processed products. 81 iced frui ts exhibi t increas~d ethylene production and respiration compal'ed to whole f, 'uits during distribution in response to tissue damage. As a result, accelerated senescence and enzymatic browning OCCUI', Recent l'esearch on minimally processed fl'uits and vegetables has mainly focused on methods to inhibit browning, due to ban on use of sulfur dioxide, In order to retard or prevent these physiological changes, val'ious al ternatives, reducing agents. acidulants, chelating agents and inol'ganic sal ts have been evaluated for use on fresh cut fl'ui ts. Al though some agents were effective replacement for sulfur dioxide. consum$\textregistered$I'S demandless use of chemical on such products. Shel~ life of minimally processed products has been extended by inhibition of metabolic reactions associated with loss of quality and by inhibition of aerobic spoilage caused by wide variety of microorganisms. Appl ication of ~I.-\ packaging, including vacuum packaging, retards the rate of respiration, prevents growth of aerobic spoilage organisms, inhibits oxidation and color deterioration. Tissue softening is another major problem in minimally processed products because enzymes re 1 a ted to ce 11 wa 11 degrada t i on are not inactivated. Various treatments have been investigated for retardation of the softening of sliced products. Some studies have concentrated on the application of an active packaging system with ~I, l. packaging and calcium infi 1 tration as possible measures to retain firmness of processed products. In my opinion, one important step for production of minimally processed frui ts wi th favorabl e color of cut surface and firm texture is the selection of better cultivar. As the view, changing tendency of fresh color by apple cultivars and relationship between the tendency and PPO activity will be discussed in the seminar. In addition to the topic, research result on quality enhancement of fresh apple slices by heat shock treatment will be introduced.

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Studies on the Antibacterial Activity of Enzymatic Hydrolyzates of Lactoferrin Derived from Bovine Colostrum (유우 Lactoferrin 효소가수분해물 항균 활성에 관한 연구)

  • Han, Su Yeon;Kim, Jong Woo
    • Korean Journal of Agricultural Science
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    • v.25 no.1
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    • pp.52-67
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    • 1998
  • The investigative research on the mammalian milk purely consisted of the physiological quality of lactoferrin was conducted to reveal the antimicrobial ativity of specifically functional foods with antibiotic characteristics as a basic data in food manufacturing. Bovine lactoferrin were isolated from raw milk samples, and was digested with pepsin, trypsin and chymotrypsin. It was necessary then to separate and purify lactoferrin from bovine raw milk, and in order to analyze the antimicrobial activity of the enzyme-treated bovine lactoferrin in their required quantitative fraction. Afterwards Escherichia coli and Staphylococcus aureus was incubated in it. It was that investigated to enzyme-treated fractions molecular weight and the peptide fragment with antimicrobial effect. 1. The purity of enzyme-treated bovine lactoferrin(BLF) was tested by SDS-PAGE. As a results of 12% SDS-PAGE assay, pepsin-treated LF did not exhibited band until if reaches 14 KDa, while trypsin and chymotrypsin treated LF, known to contain the non-digestive lactoferrin exhibited band at a molecular weight of 33 KDa. 2. Bovine lactoferrin was sucessfully purified through the use of Sephadex G-50 Column. In order to assay LF through the Sephadex G-50 column chromatography, the digestive bovine lactoferrin (BLFs) was eluted with a linear gradient of 0.05% Tris-HCl. When the gel-filtration analysis, pepsin, trypsin and chymotrypsin treatments of BLF fragments was showed 2, 3, and 2 peak, respectively. The results of the HPLC analysis confirmed that had a non-digestive lactoferrin receptor, and trypsin and chymotrypsin treated BLFs has an antimicrobial effect. 3. To measure the strength of the antimicrobial effect of enzyme treated lactoferrin it was compared to the antimicrobial activity taking place at the incubated Escherichia coli and Staphylococcus aureus. This might explain the resistance of the microorganisms for peptide fragment. The pepsin-treated of bovine lactoferrin was markedly reduced by incubation of the cells. Trypsin-treated of BLF was similar to chymotrypsin-treated of BLF. However, trypsin and chymotrypsin treatments of BLFs were showed the antimicrobial effect until eight hours incubation for native bovine lactoferrin. Therefore the enzyme-treated lactoferrin have an antimicrobial effect even non-digestive lactoferrin. 4. The digestive bovine lactoferrin fragments assay was carried out by the use of Sephadex G-50 column chromatography and SDS-PAGE. The pepsin and chymotrypsin-treated fragments has a low molecular weight and trypsin-treated lactoferrin was only showed a band. It was described that characteristics of digestive protein. It appeared that there may be a relation between virulence and resistance to enzyme-treated BLF.

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Physicochemical Characteristics of ${\beta}-Glucan$ Isolated from Barley (보리로부터 분리한 ${\beta}-Glucan$의 이화학적 특성)

  • Kim, Sung-Ran;Choi, Hee-Don;Seog, Ho-Moon;Kim, Sung-Soo;Lee, Young-Tack
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1164-1170
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    • 1999
  • The physicochemical characteristics of ${\beta}-glucan$ isolated from waxy and non-waxy barley were investigated. The hull-less waxy and non-waxy barley containing 6.5% and 5.3% of total ${\beta}-glucan$ respectively, were used as a starting material. The yield and ${\beta}-glucan$ content of crude ${\beta}-glucan$ from waxy barley was 5.54% and 62.9%, respectively, and those were higher than 3.34% and 59.2% from non-waxy barley. The crude ${\beta}-glucan$ purified with selective precipitation and enzymatic treatment to obtain the ${\beta}-glucan$ isolate of high purity (>99%). The total yield of purified ${\beta}-glucan$ from waxy and non-waxy barley was 4.46% and 2.59%, respectively. The surface appearance of the purified ${\beta}-glucan$ by scanning electron microscopy (SEM) showed randomly entangled multi-net structure of ${\beta}-glucan$ microfibrils. The melting temperature of ${\beta}-glucan$ from waxy and non-waxy barley measured by differential scanning calorimetry (DSC) was $184.6^{\circ}C$, and $180.3^{\circ}C$, respectively. DSC endotherm of ${\beta}-glucan$ solution showed 2 peaks near $68^{\circ}C$ and $84^{\circ}C$. Enthalpy of phase transition was higher in non-waxy ${\beta}-glucan$ than waxy ${\beta}-glucan$, and the intrinsic viscosity of ${\beta}-glucan$ solution from waxy barley was higher than that of non-waxy ${\beta}-glucan$. The pasting viscosity of barley starch with the purified ${\beta}-glucan$ determined by Rapid Visco-Analyzer was higher than that of barley starch without ${\beta}-glucan$, and the effect of ${\beta}-glucan$ on increasing the paste viscosity was greater in non-waxy barley starch.

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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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