• Title/Summary/Keyword: carbohydrate specificity

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Tumor Markers in Serum and Ascites in the Diagnosis of Benign and Malignant Ascites

  • Zhu, Fang-Lai;Ling, An-Sheng;Wei, Qi;Ma, Jie;Lu, Gang
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.2
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    • pp.719-722
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    • 2015
  • Objective: To evaluate the values of 4 tumor markers in serum and ascites and their ascites/serum ratios in the identification and diagnosis of benign and malignant ascites. Materials and Methods: A total of 76 patients were selected as subjects and divided into malignant ascites group (45 cases) and benign ascites group (31 cases). Samples of ascites and serum of all hospitalized patients were collected before treatment. The levels of carcinoembryonic antigen (CEA), alpha fetoprotein (AFP), cancer antigen 125 (CA125) and carbohydrate antigen 19-9 (CA19-9) were detected by chemiluminescence (CLIA). Results: CEA, AFP and CA19-9 in both serum and ascites as well as CA125 in ascites were evidently higher in the malignant ascites group than in the benign ascites group (P<0.01). Malignant ascites was associated with elevated ascites/serum ratios for AFP and CA125 (P<0.01). The areas under receiver operating characteristic (AUROCs) of CEA and CA125 in ascites and the ratios of ascites/serum of AFP, CEA, CA125 and CA19-9 were all >0.7, suggesting certain values, while those of ascites CA19-9 and serum CEA were 0.697 and 0.629 respectively, indicating low accuracy in the identification and diagnosis of benign and malignant ascites. However, the AUROCs of the remaining indexes were <0.5, with no value for identification and diagnosis. Compared with single index, the sensitivity of combined detection increased significantly (P<0.05), in which the combined detection of CEA, CA19-9 and CA125 in ascites as well as the ratio of ascites/serum of CEA, CA19-9, CA125 and AFP had the highest sensitivity (98.4%) but with relevantly low specificity. Both sensitivity and specificity of combined detection should be comprehensively considered so as to choose the most appropriate index. Conclusions: Compared with single index, combined detection of tumor markers in serum and ascites can significantly improve the diagnostic sensitivity and specificity.

Isolation and Carbohydrate Binding Specificity of a Lectin from the Hemolymph of Coastal Crab Macrophthalmus Japonicus (해양칠게 혈림프로부터 렉틴의 분리 및 당 결합특성)

  • Kim, Ha-Na;Chung, Woo-Hyuk;Bae, Chan-Hyung;Hwang, Kwang-Woo;Kim, Ha-Hyung
    • YAKHAK HOEJI
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    • v.50 no.3
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    • pp.166-171
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    • 2006
  • A calcium-dependent sialic acid-binding lectin has been isolated by thyroglobulin-affinity chromatography from the coastal crab Macrophthalmus Japonicus. This lectin, Macrophthalmus Japonicus lectin (MJL), was eluted with 50mM Tris-HCl, 0.3 M NaCl, 10 mM EDTA, and the recovery yield from the crude protein extract was about 5.6%. The molecular weight of MJL was estimated as 65 kDa in SDS-PAGE both under reducing and non-reducing conditions. MJL induced an agglutination reaction in rabbit, rat, and mouse erythrocytes, but not in human ABO types. This activity was effectively inhibited by sialoglycoproteins such as fetuin, bovine submaxillary mucin, and thyroglobulin.

Expression of Carboxypeptidase Z cDNA from Absidia zychae in saccharomyces cerevisiae and its characteristics (Saccharomyces cerevisiae에서 Absidia zychae 의 Carboxypeptidase A cDNA 의 발현과 특성)

  • 이병로;김종화
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.150-155
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    • 1995
  • Carboxypeptidase Z(CPZ) cDNA of Absidia zychae was experssed in Saccharomyces cerevisiae. The expressed CPZ(YCPZ) was secreted about 30 mg/l into the medium and has a little higher molecular weight than the wild type CPZ in SDS-PAGE. By the result of N-terminal amino acid sequencing, YCPZ has additional 15 amino acids residues in N-terminus of CPZ. But YCPZ shows no difference with CPZ in enzyme activity and substrate specificity. For the identification of processing mechanism of YCPZ, 36-Arg was changed to 36-Thr by site specific mutagenesis. Mutant YCPZ does not processed at 36-Thr. It was, therefore, concluded that the YCPZ was processed by KEX2. According to endo F treatment, high amount of carbohydrate was N-glycosylated in YCPZ.

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Enzymatic Characteristics and Applications of Microbial Chitin Deacetylases (미생물 Chitin Deacetylase의 특성과 응용)

  • Kuk Ju-Hee;Jung Woo-Jin;Kim Kil-Yong;Park Ro-Dong
    • Microbiology and Biotechnology Letters
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    • v.33 no.1
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    • pp.9-15
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    • 2005
  • Chitin deacetylase (CDA; EC 3.5.1.41) catalyzes the hydrolysis of N-acetamide bonds of chitin, converting it to chitosan. Chitosan has several applications in areas such as biomedicine, food ingredients, cosmetics, pharmaceuticals, and agriculture. In this paper, occurrence, assay and purification protocols, enzymatic characteristics, substrate specificity, and mode of action of microbial CDAs have been described. Several lines of evidence have substantiated the biological roles involved in cell wall formation and plant-pathogen interactions for fungal CDAs. The gene structure of CDAs has been compared with other family 4 carbohydrate esterases which deacetylate a wide variety of acetylated poly/oligo-saccharides. The use of CDAs for the conversion of chitin to chitosan, in contrast to the presently used chemical procedure, offers the possibility of a controlled, non-degradable process, resulting in the production of well-defined chitosan oligomers and polymers. Insect pathogen that can secrete high levels of chitin-metab­olizing enzymes including CDA can be a possible alternative for new pest management tools.

Roles of Carbohydrate-Binding Module (CBM) of an Endo-β-1,4-Glucanase (Cel5L) from Bacillus sp. KD1014 in Thermostability and Small-Substrate Hydrolyzing Activity

  • Lee, Jae Pil;Shin, Eun-Sun;Cho, Min Yeol;Lee, Kyung-Dong;Kim, Hoon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2036-2045
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    • 2018
  • An endo-${\beta}$-1,4-glucanase gene, cel5L, was cloned using the shot-gun method from Bacillus sp.. The gene, which contained a predicted signal peptide, encoded a protein of 496 amino acid residues, and the molecular mass of the mature Cel5L was estimated to be 51.8 kDa. Cel5L contained a catalytic domain of glycoside hydrolase (GH) family 5 and a carbohydrate-binding module family 3 (CBM_3). Chromatography using HiTrap Q and CHT-II resulted in the isolation of two truncated forms corresponding to 50 (Cel5L-p50) and 35 kDa (Cel5L-p35, CBM_3-deleted form). Both enzymes were optimally active at pH 4.5 and $55^{\circ}C$, but had different half-lives of 4.0 and 22.8 min, respectively, at $70^{\circ}C$. The relative activities of Cel5L-p50 and Cel5L-p35 for barley ${\beta}$-glucan were 377.0 and 246.7%, respectively, compared to those for carboxymethyl-cellulose. The affinity and hydrolysis rate of pNPC by Cel5L-p35 were 1.7 and 3.3 times higher, respectively, than those by Cel5L-p50. Additions of each to a commercial enzyme set increased saccharification of pretreated rice straw powder by 17.5 and 21.0%, respectively. These results suggest CBM_3 is significantly contributing to thermostability, and to affinity and substrate specificity for small substrates, and that these two enzymes could be used as additives to enhance enzymatic saccharification.

Tumour Markers in Peritoneal Washing Fluid - Contribution to Cytology

  • Yildirim, Mustafa;Suren, Dinc;Yildiz, Mustafa;Alikanoglu, Arsenal Sezgin;Kaya, Vildan;Doluoglu, Suleyman Gunhan;Aydin, Ozgur;Yilmaz, Necat;Sezer, Cem;Karaca, Mehmet
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.1027-1030
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    • 2013
  • Background: Peritoneal washing cytology (PWC) that shows the microscopic intra-peritoneal spread of gynaecologic cancers is not used in staging but is known as prognostic factor and effective in planning the intensity of the therapy. False negative or false positive results clearly affect the ability to make the best decision for therapy. In this study we assessed levels of tumour markers, carcinoembryonic antigen (CEA), cancer antigen 125 (CA-125) and carbohydrate antigen (CA19-9), in peritoneal washing fluid to establish any possible contribution to the peritoneal washing cytology in patients operated for gynaecologic cancer. Materials and Methods: Preoperative tumour markers were studied in serum of blood samples obtained from the patients for preoperative evaluation of a gynaecologic operation. In the same group peritoneal tumour markers were studied in the washing fluid obtained for intraoperative cytological evaluation. Results: This study included a total of 94 patients, 62 with malignant and 32 with benign histopathology. The sensitivity of the cytological examination was found to be 21% with a specificity of 100%. When evaluated with CEA the sensitivity of the cytological examination has increased to 37%. Conclusions: In addition to examination of PWC, the level of CEA, a tumour marker, in peritoneal washing fluid can make a diagnostic contribution. Determining the level of CEA in peritoneal washing fluid will be useful in the management of gynaecologic cancers.

Biochemical Properties of Eggplant Fruit Lectin. (가지 열매 lectin의 생화학적 성질)

  • Roh, Kwang-Soo
    • Journal of Life Science
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    • v.18 no.3
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    • pp.350-356
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    • 2008
  • Biochemical characterization including hemagglutination of erythrocytes, molecular weight, optimum temperature, thermal stability, optimum pH, carbohydrate specificity, and inhibitory effect of metal ion were studied in lectin of eggplant (Solanum melongena L.) fruit prepared by ammonium sulfate fractionation and affinity chromatography. This lectin was agglutinated by trypsin-treated rat blood erythrocyte. The molecular weight of this lectin by SDS-PAGE was estimated to be approximately 19.3 kDa of a single band. This lectin has no activity by 7 carbohydrates containing D-glucose. The optimum range of temperature and pH were $10-20^{\circ}C$ and pH 6.2-7.2, respectively. This lectin was relatively stable at $20-70^{\circ}C$. And the activity of this lectin was not inhibited by $Ca^{2+},\;Co^{2+},\;Cu^{2+},\;Fe^{2+},\;Mg^{2+}$, and $Mn^{2+}$.

Characteristics and Cancerostatic Activity of the Starfish Lectin (별불가사리 렉틴의 특성 및 암 세포 성장저해 효과)

  • Jeune, Kyung-Hee;Park, Chae-Soo;Park, Won-Hark;Choi, Soo-Jeong;So, Myung-Suk;Chung, See-Ryun
    • YAKHAK HOEJI
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    • v.41 no.4
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    • pp.421-432
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    • 1997
  • A new lectin was partially purified from starfish,Asterina pectinifera by means of physiological saline extraction, salt fractionation, ion exchange chromatography and hy droxyapatite chromatography, and it was named APL. The biochemical properties of the APL were characterized. In addition, its effects on lymphocyte mitogenicity and cancer cell agglutinability were tested. The APL agglutinated nonspecifically human erythrocytes and rabbit blood cells. Agglutinability was decreased to 30% of control activity below pH 5 and above pH 9 and was relatively unstable at increasing temperatures above 60$^{\circ}C$. The activity was reduced by addition of two kinds of metal ions, $Ba^{2+},\;Mn^{2+}$ and chelating agent, EDTA. APL was proved to be glycoproteins containing 9% sugars. For carbohydrate specificity, it was found that the activity of APL was inhibited by D(+)-glucosamine, D(+)-galactosamine, stachyose, N-acetyl-galactosamine and methyl-${\alpha}$-D-galactopyranoside among 35 sugars tested. In amino acid composition, the contents of acidic amino acids such as aspartic acid and glutamic acid were relatively high. This result suggest that the isoelectric point would be in a lower range. APL was found that it promotes the division of human lymphocytes. APL was proved to be a potent agglutinin for cancer cells such as HeLa, L929 and L1210 cells. Significant changes on the HeLa cell surfaces affected by APL were observed under the electron microscope.

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Diagnostic Role of F-18 FDG PET/CT in the Follow-up of Patients with Colorectal Cancer: Comparison with Serum CEA, CA 19-9 Levels and Computed Tomography (대장암 치료 후 추적 검사로서 F-18 FDG PET/CT의 역할: 혈청 CEA, CA 19-9 및 Computed Tomography와의 진단 성능 비교)

  • Kang, Sung-Min;Song, Bong-Il;Lee, Hong-Je;Seo, Ji-Hyoung;Lee, Sang-Woo;Yoo, Jeong-Soo;Ahn, Byeong-Cheol;Lee, Jae-Tae;Choi, Kyu-Suk;Jun, Soo-Han
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.2
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    • pp.120-128
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    • 2009
  • Purpose: Early detection of recurrence is an important factor for long term survival of patients with colorectal cancer. Measurement of serum levels of CEA, CA 19-9, CT and PET/CT has been commonly used in the postoperative surveillance of colorectal cancer. The purpose of this study was to compare the diagnostic ability of PET/CT, tumor marker and CT for recurrence in colorectal cancer patients after treatment. Materials and Methods: F-18 FDG PET/CT imaging was performed in 189 colorectal cancer patients who underwent curative surgical resection and/or chemotherapy. Measurement of serum levels of CEA, CA 19-9 and CT imaging were performed within 2 months of PET/CT examination. Final diagnosis of recurrence was made by biopsy, radiologic studies or clinical follow-up for 6 months after each study. Results: Overall sensitivity, specificity of PET/CT was 94.7%, 91.1%, while those of serum CEA were 44.7% and 97.3%, respectively. Sensitivity and specificity were 94.2%, 90.4% for PET/CT and better than those of combined CEA and CA 19-9 measurement(52.1%, 88.5%) in 174 patients measured available both CEA and CA 19-9 data. In 115 patients with both tumor markers and CT images available, PET/CT showed similar sensitivity but higher specificity(92.9%, 91.3%) compared to combination of tumor markers and CT images(92.9%, 74.1%). Conclusion: PET/CT was superior for detection of recurred colorectal cancer patients compared with both CEA, CA 19-9, and even with combination of both tumor markers and CT. Therefore PET/CT could be used as a routine surveillance examination to detect recurrence or metastasis of colorectal cancer.

Characterization of Laccase Purified from Korean Trametes hirsuta S1 (한국산 흰구름버섯(Trametes hirsuta S1)으로부터 정제된 Laccase의 특성)

  • Lim, Hyung-Seon;Yoon, Kyung-Ha
    • The Korean Journal of Mycology
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    • v.32 no.2
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    • pp.112-118
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    • 2004
  • Laccase produced by Trametes hirsuta S1 isolated from Korea was partially purified and characterized using ultrafiltration, anion exchange chromatography and affinity chromatography. The laccase was produced as the predominant extracellular enzyme during primary metabolism. Neither lignin peroxidase nor veratryl alcohol oxidase (VAO) were detected in the culture fluid. Addition of 2,5-xylidine enhanced 4-fold laccase production. Purified laccase was a single polypeptide having a molecular mass of approximately 66 kDa, as determined by SDS-polyacrylamide gel electrophoresis, and carbohydrate content of 12%. $K_{m}\;and\;V_{max}$ values for laccase with ABTS [2,2-azinobis (3-ethylbenzthiazoline 6-sulfonic acid)] as a substrate (Lineweaver-Burk plot) was determined to $51.2\;{\mu}M\;and\;56.8\;{\mu}mole$, respectively. The optimal pH for laccase activity was found to be 3.0. The enzyme was very stable for 1 hour at $50^{\circ}C$. Half-life ($t_{1/2}$) of the enzyme was about 20 min at $70^{\circ}C$. Spectroscopic analysis of purified enzyme indicated that the enzyme was typical of copper-containing protein. Substrate specificity and inhibitor studies for laccase also indicated to be a typical fungal laccase. The N-terminal amino acid sequence of the T. hirsuta S1 laccase showed 100% of homology to those of laccase from C. hirsutus.