• Korean Journal of Microbiology
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• v.48 no.2
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• pp.73-78
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• 2012

We previously reported that the truncated Cel44C-$Man26A_{P558}$ ${\beta}$-glycosyl hydrolase protein exhibits multifunctional activities, including cellulase, xylanase, and lichenase. DNA shuffling of the truncated Cel44C-$Man26A_{P558}$ enzyme was performed to enhance the enzymatic activity of the multifunctional ${\beta}$-glycosyl hydrolase. Two mutant enzymes, M2Cel44C-$Man26A_{P558}$ that carries one mutation (P438A) and M21Cel44C-$Man26A_{P558}$ that carries two mutations (A273T and P438A) were obtained. The enzymatic activity of the M21Cel44C-$Man26A_{P558}$ double mutant was lower than enzymatic activity of the single mutant (M2Cel44C-$Man26A_{P558}$). However, both mutants displayed the enhancements in their enzyme activities that were ${\approx}1.3$- to 2.2-fold higher than the original enzymatic activity in Cel44C-$Man26A_{P558}$. In particular, the mutant M2Cel44C-$Man26A_{P558}$ exhibited an approximate 1.5- to 2.2-fold increase in the cellulase, xylanase, and lichenase activities in comparison with the control (Cel44C-$Man26A_{P558}$). The optimum cellulase, linchenase, and xylanase activities of ${\beta}$-glycosyl hydrolase were observed at pH 7.0, pH 7.0 and pH 6.0, respectively. These results, therefore, suggest that the amino acid residue Ala438 plays important roles in the enhancement of the activity of multifunctional ${\beta}$-glycosyl hydrolase.

• Journal of Animal Environmental Science
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• v.18 no.3
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• pp.191-200
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• 2012

This study was conducted to investigate the effects of application of cattle slurry on forage productivity and environmental pollution in rice paddy land. Cropping systems used in this study consisted of two designs, such as whole crop rice applied with standard fertilizer (MWRS) and whole crop rice applied with cattle slurry (MWRC). The field experiments were conducted on the clay loam at Backsanmyun, Kimje, Chunlabukdo province in Korea for three years (May 2006 to Apr. 2009). This study was arranged in completely randomized design with three replicates. The yield of dry matter of whole crop rice in MWRS was similar as compared with that of MWRC. The contents of crude protein of whole crop rice in MWRS significantly increased as compared with those of MWRC (P<0.05). However, the contents of acid detergent fiber (ADF), neutral detergent fiber (NDF) and total digestible nutrients (TDN) of whole crop rice were hardly influenced by cattle slurry application. The pH, and contents of T-N, $P_2O_5$ and organic matter (OM) in soil samples collected at the end of the experiment increased as compared with those at the beginning of the experiment. After the end of experiment, the concentrations of exchangeable cations (Ca, Na, Mg and K) in soil samples collected at the end of the experiment were remarkably higher than those at the beginning of the experiment (P<0.05). The concentrations of $NO_3$-N and $NH_4$-N in discharge water in MWRS, MWRC and DWBRC were higher in 2008 than those in 2007. But the concentration of $PO_4$-P in discharge water was hardly influenced by the cropping system during experimental period. The concentrations of $NO_3$-N, $NH_4$-N and $PO_4$-P in leaching water were hardly influenced by application of cattle slurry.

• Journal of Yeungnam Medical Science
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• v.11 no.1
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• pp.42-48
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• 1994

Among the erythrocyte membrane defects, hereditary spherocytosis is the most common. The erythrocyte membrane defect results from a deficiency of spectrin, the most important structural protein in red cell. Hereditary spherocytosis often presents with hemolytic anemia, jaundice, moderate splenomegaly. Diagnosis is established by the presence of spherocytes in the peripheral blood, reticulocytosis, an increased osmotic fragility, and a negative Coombs test. In children, splenectomy is usually performed after age 6 years but can be done at a younger age if warranted by the severity of the anemia and the need for frequent transfusions. In the period December 1987 to Agust 1993, 9 patients with hereditary spherocytosis underwent splenectomy and the following results were obtained. 1. Nine patients were comprised of five males and four females. 2. Five patients(55.6%) had been admitted to our hospital during age 6-10 years. 3. Four of the nine patients had autosomal dominant inheritance with variable expression. The other five patients had no known inheritance. 4. The diagnosis of the spherocytosis was based on the increased osmotic fragility and increased autohemolysis of the erythrocytes, as well as on the appearance of spherocytes in the peripheral blood smear. 5. In all cases splenectomy was performed. Two patients had concomitant gall stones and choledocholithiasis, respectively. One patient with concomitant gall stones underwent simultaneous cholecystectomy and splenectomy. The other patient associated with choledocholithiasis underwent splenectomy, cholecystectomy, choledocholithotomy, and T-tube drainage. 6. Complete hematologic recovery was obtained by the splenectomy in all cases. 7. Postoperative complication was not occurred.

• The Korean Journal of Nuclear Medicine Technology
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• v.15 no.1
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• pp.117-120
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• 2011

Purpose: There are 3 warnings for Glucagon tests. First, EDTA tubes that already contain Aprotinin must be used for plasma collection. Second, for freezer storage of centrifuged plasma, glass tubes must be used. Last, glass tubes must be used for testing procedure. So we compared the glucagon results of next 3 situation to those of control group. First, We compared to results by tubes without Aprotinin and with aprotinin. Second, we compared to results by tubes(plastic vs glass) for plasma storage. Third, we compared to results by tubes(plastic vs glass) for testing. We tried to evaluate the results of the 3 different condition. Materials and Methods: 40 healthy adults were studied with normal results on the general medical check up and laboratory tests. We compared the results of 3 different condition belows: Blood were collected in EDTA tube containing aprotinin and plasma was stored in the glass tube for 3 days in a freezer and results were obtained by tests in the glass tubes. Results from EDTA plasma without aprotinin, results from platic tubes for freezer stroage, results from plastic tube when testing. Simple linear regression analysis and paired t-test using SPSS were done for statistical analysis. Commercial glucagon kit(RIA-method)which made by Siemens company were used. Results: Correlation coefficient between results of EDTA tubes with Aprotinin vs without Aprotinin was r=0.783 (p=0.064). Result of specimen in plastic tubes stored 3 days in a freezer showed lower value compared to those in glass tube(r=0.979, p=0.005). Also, results of testing in plastic tubes showed lower values than those testing in glass tubes. (r=0.754, p<0.001). Conclusion: It is recommended for glucagon determination to use EDTA tube with Aprotinin which is a inhibitor of protein breakdown enzyme. Results of plastic tube when storage and testing showed lower value than those of glass tubes, so it is recommended to store and test in glass tubes.

• Journal of Chest Surgery
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• v.39 no.11 s.268
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• pp.828-837
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• 2006

Background: Tissue hypoxia is characteristic of many human malignant neoplasm, and hypoxia inducible factor-1(HIF-1) plays a pivotal role in essential adaptive response to hypoxia, and activates a signal pathway for the expression of the hypoxia-regulated genes, resulting in increasing $O_2$ delivery or facilitating metabolic adaptation to hypoxia. Increased level of HIF-$1{\alpha}$ has been reported in many human malignancies, but in non-small cell lung carcinoma the influence of HIF-$1{\alpha}$ on tumor biology, including neovascularization, is not still defined. In present study the relationship of HIF-$1{\alpha}$ expression on angiogenetic factors, relationship between the tumor proliferation and HIF-$1{\alpha}$ expression, interaction of HIF-$1{\alpha}$ expression and p53, and relationship between HIF-$1{\alpha}$ expression and clinico-pathological prognostic parameters were investigated. Material and Method: Archival tissue blocks recruited in this study were retrieved from fifty-nine patients with primary non-small cell lung carcinoma, who underwent pneumonectomy or lobectomy from 1997 to 1999. HIF-$1{\alpha}$, VEGF(vascular endothelial growth factor), and p53 protein expression and Ki-67 labeling index in tumor tissues were evaluated, using a standard avidin-biotin-peroxidase complex(ABC) immunohistochemistry. Relationship between the HIF-$1{\alpha}$ expression and VEGF, p53 overexpression and correlation between the HIF-$1{\alpha}$ expresseion and Ki-67 index were analyzed. Clinico-pathologic prognostic parameters were also analyzed. Result: HIF-$1{\alpha}$ expression in cancer cells was found in 24 of 59 cases of non-small cell lung carcinoma(40.7%). High HIF-$1{\alpha}$ expression was significantly associated with several pathological parameters, such as pathological TMN stage(p=0.004), pT stage(p=0.020), pN stage (p=0.029), and lymphovascular invasion(p=0.019). High HIF-$1{\alpha}$ expression was also significantly associated with VEGF immunoreactivity(p<0.001), and aberrant p53 expression(p=0.040). but was marginally associated with Ki-67 labeling index(p=0.092). The overall 5-year survival rate was 42.3%. The survival curve of patients with a high HIF-$1{\alpha}$ expression was worse than that of patients with low-expression(p=0.002). High HIF-$1{\alpha}$ expression was independent unfavorable factors with a marginal significance in multivariate analysis performed by Cox regression. Conclusion: It is suggested that high HIF-$1{\alpha}$ expression may be associated with intratumoral neovascularization possibly through HIF-VEGF pathway, and high HIF-$1{\alpha}$ expression could be associated with lymph node metastasis and post operative poor prognosis in patients with non-small cell lung carcinoma.

• The Korean Journal of Nuclear Medicine
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• v.28 no.1
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• pp.98-105
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• 1994

The search for tumor-avid agents for use in nuclear medicine imaging or therapy is a field of ongoing importance. Metallothionein (MT) is an intracellular protein that binds many metals with isotopes having imaging or radiotherapeutic potential. The purpose of the study was to determine whether uptake of radioisotopes that bind to MT is increased in tumor. We measured the uptake of Cd-109 and Ga-67 in tumor and normal tissues of sarcoma-bearing mice. Tumors were grown subcutaneously in female Balb/C mice from cultured Balb/3T3 cells transformed by the Moloney murine sarcoma virus (MMSV). When the tumors reached about 1 cm in diameter, mice were injected subcutaneously with Cd-109 and Ga-67. Eighteen and seventy-two hours later, the mice were sacrified. Organs and tissues were removed, weighed, and activity per mg tissue determined by gamma well-counting. Uptake of Cd-109 by MMSV tumors exceeded that by normal tissues examined, with the exception of liver and kidney (the organs known to be richest in MT). The tumor-to-tissue ratios of uptake for Cd-109 were far greater than those for Ga-67 for many normal tissues of great importance in terms of background activity (bone, intestine, fat, muscle, and blood). We concluded that metals that bind to MT may be useful for oncologic imaging or rediotherapy of cancer.

• Journal of Oral Medicine and Pain
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• v.34 no.3
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• pp.261-276
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• 2009

Lactacystin, a microbial natural product synthesized by Streptomyces, has been commonly used as a selective proteasome inhibitor in many studies. Proteasome inhibitors is known to be preventing the proliferation of cancer cells in vivo as well as in vitro. Furthermore, proteasome inhibitors, as single or combined with other anticancer agents, are suggested as a new class of potential anticancer agents. This study was undertaken to examine in vitro effects of cytotoxicity and growth inhibition, and the molecular mechanism underlying induction of apoptosis in SCC25 human tongue sqaumous cell carcinoma cell line treated with lactacystin. The viability of SCC25 cells, human normal keratinocytes (HaCaT cells) and human gingiva fibroblasts (HGF-1 cells), and the growth inhibition of SCC25 cells were assessed by MTT assay and clonogenic assay respectively. The hoechst staining, hemacolor staining and TUNEL staining were conducted to observe SCC25 cells undergoing apoptosis. SCC25 cells were treated with lactacystin, and Western blotting, immunocytochemistry, confocal microscopy, FAScan flow cytometry, MMP activity, and proteasome activity were performed. Lactacystin treatment of SCC25 cells resulted in a time- and does-dependent decrease of cell viability and a does-dependent inhibition of cell growth, and induced apoptotic cell death. Interestingly, lactacytin remarkably revealed cytotoxicity in SCC25 cells but not normal cells. And tested SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA contents, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, the up-regulation of Bax, and the activation of caspase-7, caspase-3, PARP, lamin A/C and DFF45 (ICAD). Flow cytometric analysis revealed that lactacystin resulted in G1 arrest in cell cycle progression which was associated with up-regulation in the protein expression of CDK inhibitors, $p21^{WAF1/CIP1}$ and $p27^{KIP1}$. We presented data indicating that lactacystin induces G1 cell cycle arrest and apoptois via proteasome, mitochondria and caspase pathway in SCC25 cells. Therefore our data provide the possibility that lactacystin could be as a novel therapeutic strategy for human tongue squamous cell carcinoma.

• Tuberculosis and Respiratory Diseases
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• v.59 no.4
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• pp.406-412
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• 2005

Background : Recently, two commercialized whole-blood assays, $QuantiFERON^{(R)}-TB$ Gold (QFT) and T $SPOT-TB^{(R)}$ (SPOT), which measure the $IFN-{\gamma}$ released in the whole blood after being incubation with mycobacterial antigens, were approved for the diagnosis of a latent tuberculosis infection (LTBI). However, there is data on whether or not the previously used PPD skin tests (TST) have any influence on the diagnostic ability of these ex-vivo $IFN-{\gamma}$ assays. Methods : Forty-six 15 year-old students who did not appear to be infected with Mycobacterium tuberculosis were enrolled in this study. The peripheral blood was collected and used for two $IFN-{\gamma}$ assays. The $IFN-{\gamma}$ assays and TST were performed at the baseline ($1^{st}$). The TST was repeated two months later ($2^{nd}$), and the $IFN-{\gamma}$ assays were repeated two ($2^{nd}$) and four months ($3^{rd}$) later only in those subjects who had negative results at the baseline in both the $IFN-{\gamma}$ assays and TST. An induration size > 10 mm was considered to be positive in the TST. Results : The mean TST value was $3.1{\pm}5.4mm$ (range: 0-20). Of the 46 subjects examined, 13 subjects (28.3%) showed positive results in the two-step TST. Nine (19.6%) were SPOT-positive and only one (2.2%) was QFT-positive. The $2^{nd}$ and $3^{rd}$ QFT were carried out in 23 and 25 all-negative subjects, respectively, and all showed negative results. The $2^{nd}$ SPOT was performed in 23 subjects and only one (4.3%) showed a weak-positive result. Conclusion : Even though there were some discrepancies in the results of the two ex-vivo $IFN-{\gamma}$ assays, it appears that their results were not influenced by a previous TST carried out in two or four months earlier.

• Food Science and Preservation
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• v.23 no.6
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• pp.811-818
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• 2016

The purpose of this study was to investigate changes in the amino acid content and physicochemical properties of Cheonggukjang prepared by using various soybean cultivars (Daewon, Deapung, Seadanbeak, and Taekwang) and a functional microorganism (Bacillus subtilis HJ18-9). These soybeans were conventional Cheonggukjang (control) and Cheonggukjang fermented with Bacillus subtilis HJ18-9 (treated). The moisture contents of steamed, control, and treated soybean were 62.45~67.12%, 63.28~67.14%, and 64.50~66.87%; amino-type nitrogen contents were 6.53~24.25 mg%, 27.63~122.09 mg%, and 37.29~133.48 mg%; and ammonia-type nitrogen contents were 26.92~47.95 mg%, 45.45~156.36 mg%, and 28.02~121.13 mg%, respectively. The umami taste associated with several amino acids (aspartic acid and glutamic acid) in Cheonggukjang was lower than that for steamed soybeans, while the bitter taste from amino acids (methionine, valine, isoleucine, leucine, and phenylalanine) was higher than that for steamed soybeans. The result of sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that the molecular weight of steamed soybeans was less than 100 kDa, while control and treated groups showed low molecular weights below 34 kDa, confirming their protein hydrolysis to small molecular weight. These results are information for developing functional fermented soybean paste and diversification using soybean cultivars.

• 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.