• Title/Summary/Keyword: Aga2p

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Influence of Long-term Fertilization on soil Enzymes Activity in Normal Paddy Soil (퇴비(堆肥) 및 비료(肥料) 장기연용(長期連用)이 토양내(土壤內) 효소활성(酵素活性)에 미치는 영향(影響))

  • Cho, Kang-Jin;Jung, Yeun-Tae;Choi, Jyung
    • Applied Biological Chemistry
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    • v.32 no.2
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    • pp.109-115
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    • 1989
  • This study was aimed to find out the influence of long-term fertilization for 21 years on soil enzyme activities in the silty clay loam textured normal paddy soil. Total urease activity (TUA) and the microbial urease activity (MUA) were shown to be changed significantly, but the accumulated urease activity (AUA) was similar within trial plots. Especially the MUA of the plots annually applied N.P.K. fertilizers with compost and N.P.K. fertilizers with silicate fertilizer were the highest among plots. The total L-glutaminase activity (TGA) and the accumulated L-glutaminase activity(AGA) were changed significantly among trial plots, but the microbial L-glutaminase activity (MGA) was not. By the simple correlation analysis, it was shown that the TGA and the AGA correlated highly significant to available phosphorus available $SiO_2$ content and pH. Addition of the toluene to the incubation mixture did not markedly affect the activity of phosphatase, but the difference of phosphatase activity among plots was significant. By the simple correlation analysis, it was shown that the phosphatase activity ; correlated highly significant to pH, available $SiO_2$, available phosphorus and exchangeable calcium in soils.

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Baseline Stimulated Thyroglobulin Level as a Good Predictor of Successful Ablation after Adjuvant Radioiodine Treatment for Differentiated Thyroid Cancers

  • Fatima, Nosheen;uz Zaman, Maseeh;Ikram, Mubashir;Akhtar, Jaweed;Islam, Najmul;Masood, Qamar;Zaman, Unaiza;Zaman, Areeba
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.15
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    • pp.6443-6447
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    • 2014
  • Background: To determine the predictive value of the baseline stimulated thyroglobulin (STg) level for ablation outcome in patients undergoing adjuvant remnant radioiodine ablation (RRA) for differentiated thyroid carcinoma (DTC). Materials and Methods: This retrospective study accrued 64 patients (23 male and 41 female; mean age of $40{\pm}14$ years) who had total thyroidectomy followed by RRA for DTC from January 2012 till April 2014. Patients with positive anti-Tg antibodies and distant metastasis on post-ablative whole body iodine scans (TWBIS) were excluded. Baseline STg was used to predict successful ablation (follow-up STg <2 ng/ml, negative diagnostic WBIS and negative ultrasound neck) at 7-12 months follow-up. Results: Overall, successful ablation was noted in 37 (58%) patients while ablation failed in 27 (42%). Using the ROC curve, a cut-off level of baseline STg level of ${\leq}14.5ng/ml$ was found to be most sensitive and specific for predicting successful ablation. Successful ablation was thus noted in 25/28 (89%) of patients with baseline STg ${\leq}14.5ng/ml$ and 12/36 (33%) patients with baseline STg >14.5 ng/ml ((p value <0.05). Age >40 years, female gender, PTS >2 cm, papillary histopathology, positive cervical nodes and positive TWBIS were significant predictors of ablation failure. Conclusions: We conclude that in patients with total thyroidectomy followed by I-131 ablation for DTC, the baseline STg level is a good predictor of successful ablation based on a stringent triple negative criteria (i.e. follow-up STg < 2 ng/ml, a negative DWBIS and a negative US neck).

Nucleotide Polymorphisms of Cytochrome P450 Genes in Domestic Garlic Cultivars (국내 재배마늘의 Cytochrome P450 유전자의 염기다형성 분포)

  • Kwon, Soon-Tae;Chung, Jinbo
    • Korean Journal of Plant Resources
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    • v.31 no.5
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    • pp.531-537
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    • 2018
  • This study was carried to survey distribution of the nucleotide polymorphisms in heme-binding (HB) domain, which is highly conserved region between 1,210 and 1,240 bp of cytochrome P450, in domestic garlic cultivars. 120 garlic cultivars collected from Korea were classified into seven HB domain variation based on the nucleotide sequence of the domain. Northern type garlic cultivars, collected from Kyungpook, Chungnam, Chungpook and Kangwon province, showed 51.3% of KP2 type nucleotide sequence, 5'-TTT/GGC/GGT/GGA/CGG/AGA/ATA/TGT/CCT/GGA-3' with coding amino acid FGGGRRICPG, 13.7% of KP1, 11.3% of CP, 8.8% of CM and 5% of KW2 types. Southern type cultivars, collected from Kyungnam province, showed 52.5% of KM type nucleotide sequence, 5'-TTT/GGC/GCA/GGA/CGG/AGA/ATT/TGT/CCT/GGA-3' with coding amino acid FGAGRRICPG, 22.5% of KP2, 5.0% of KW2 and 2,5% of CP type nucleotide sequence. These results showed that Korean garlics were cultivated in highly mixed condition even in the same region.

Expression of ${\alpha}$-Galactosidase Gene from Leuconostoc mesenteroides SY1 in Leuconostoc citreum

  • Park, Jae-Yong;Jeong, Seon-Ju;Lee, Ae-Ran;Park, Ji-Yeong;Jeong, Woo-Ju;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.12
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    • pp.2081-2084
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    • 2007
  • A 2.5 kb aga gene encoding ${\alpha}$-galactosidase (${\alpha}$-Gal) from Leuconostoc mesenteroides SY1 was cloned into pSJE, an E. coli-Leuconostoc shuttle vector. The recombinant plasmid, pSJEaga, was introduced into Leuconostoc citreum KCTC3526 (ATCC49370) by electroporation. Transcription level of aga was the highest in cells grown on raffinose (1%, w/v) followed by cells grown on galactose, melibiose, fructose, glucose, and sucrose. Western blot using antibodies against ${\alpha}$-Gal showed similar results to slot-blot results and enzyme activity measurements. All the results indicated that the aga was successfully expressed in L. citreum and its transcription was under the carbon catabolite repression (CCR).

Characterization of Cyclofructans from Inulin by Saccharomyces cerevisiae Strain Displaying Cell-Surface Cycloinulooligosaccharide Fructanotransferase

  • Kim, Hyun-Jin;Lee, Jae-Hyung;Kim, Hyun-Chul;Lee, Jin-Woo;Kim, Yeon-Hee;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.695-700
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    • 2007
  • The cycloinulooligosaccharide fructanotransferase (CFTase) gene (cft) from Paenibacillus macerans (GenBank access code AF222787) was expressed on the cell surface of Saccharomyces cerevisiae by fusing with Aga2p linked to the membrane-anchored protein Aga1p. The surface display of CFTase was confirmed by immunofluorescence microscopy and enzymatic assay. The optimized reaction conditions of surface-displayed CFTase were as follows; pH, 8.0; temperature, $50^{\circ}C$; enzyme amount, 30 milliunit; substrate concentration, 5%; inulin source, Jerusalem artichoke. As a result of the reaction with inulin, cycloinulohexaose was produced as a major product along with cycloinuloheptaose and cycloinulooctaose as minor products.

Molecular Cloning and Expression of a Novel Protease-resistant GH-36 $\alpha$-Galactosidase from Rhizopus sp. F78 ACCC 30795

  • Yanan, Cao;Wang, Yaru;Luo, Huiying;Shi, Pengjun;Meng, Kun;Zhou, Zhigang;Zhang, Zhifang;Yao, Bin
    • Journal of Microbiology and Biotechnology
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    • v.19 no.11
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    • pp.1295-1300
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    • 2009
  • A 2,172-bp full-length gene (aga-F78), encoding a protease-resistant $\alpha$-galactosidase, was cloned from Rhizopus sp. F78 and expressed in Escherichia coli. The deduced amino acid sequence shared highest identity (45.0%) with an $\alpha$-galactosidase of glycoside hydrolase family 36 from Absidia corymbifera. After one-step purification with a Ni-NTA chelating column, the recombinant Aga-F78 migrated as a single band of ~82 and ~210 kDa on SDS-PAGE and nondenaturing gradient PAGE, respectively, indicating that the native structure of the recombinant Aga-F78 was a trimer. Exhibiting the similar properties as the authentic protein, purified recombinant Aga-F78 was optimally active at $50^{\circ}C$ and pH 4.8, highly pH stable over the pH range 5.0-10.0, more resistant to some cations and proteases, and had wide substrate specificity (pNPG, melidiose, raffinose, and stachyose). The recombinant enzyme also showed good hydrolytic ability to soybean meal, releasing galactose of $415.58\;{\mu}g/g$ soybean meal. When combined with trypsin, the enzyme retained over 90% degradability to soybean meal. These favorable properties make Aga-F78 a potential candidate for applications in the food and feed industries.

Ki-67 Labeling Indices in 'Classic' versus 'Blastoid' Mantle Cell Lymphomas - Proposed Cutoff Values for Routine Diagnostic Workup

  • Pervez, Shahid;Haroon, Saroona;Awan, Dreema
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.15
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    • pp.6591-6594
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    • 2015
  • Background: Mantle-cell lymphoma (MCL) is a unique entity of peripheral B-cell lymphoma that has a discrete morphologic, immunologic, and genetic phenotype, with more common 'classic' and less frequent 'blastoid' and 'pleomorphic' variants, associated with an aggressive clinical course. The aim of this study was to analyze proliferation (Ki-67) indices of 'classic' (c-MCL) and 'blastoid' (b-MCL) variants of a cohort of MCL and to suggest cut off values for the Ki-67 proliferation index in these two subsets. Materials and Methods: MCL cases diagnosed over $4{\frac{1}{2}}$ years at Section of Histopathology, Department of Pathology and Laboratory Medicine, Aga Khan University Hospital, Karachi were retrieved and reviewed. Ki-67 labelling was scored and analysed. Results: A total of 90 of cases of MCL were scrutinized. Mean age ${\pm}SD$ was $60.2{\pm}12.5$ years and the male to female ratio was 4:1, with 67 (75%) cases of c-MCL and 23 (25%) cases of b-MCL. Most samples were lymph node biopsies (n=68), whereas the remainder were from various extranodal sites The mean Ki-67 proliferation index was $29.5%{\pm}14.4%$ in classic variants and $64.4{\pm}15.2%$ for the blastoid variant, the difference being statistically significant (p = 0.029). Conclusions: It was concluded that differential cut-off values of Ki-67 labeling may be used in more objective way to reliably classify MCL into classic or blastoid variants by diagnostic pathologists. We propose a < 40 proliferative index to be suggestive of c-MCL and one of > 50 for the blastoid variant.

Growth responses to growth hormone therapy in children with attenuated growth who showed normal growth hormone response to stimulation tests (성장호르몬 자극검사가 정상인 성장 장애 소아 환자에게서 성장호르몬 투여에 따른 성장속도의 변화)

  • Kim, Jae-Hyun;Chung, Hye-Rim;Lee, Young-Ah;Lee, Sun-Hee;Kim, Ji-Hyun;Shin, Choong-Ho;Yang, Sei-Won
    • Clinical and Experimental Pediatrics
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    • v.52 no.8
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    • pp.922-929
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    • 2009
  • Purpose : The aim was to investigate the clinical characteristics and responses to growth hormone (GH) therapy in children with attenuated growth who showed normal GH responses to GH stimulation tests (GHST). Methods : The study included 39 patients with height velocity (HV) of less than 4 cm/yr and normal GHST results. Clinical characteristics of patients were analyzed retrospectively. Results : Eleven were born as small for gestational age (SGA) and 28 as appropriate for age (AGA). In the SGA group, the standard deviation score (SDS) of age and height measured at their first visit was significantly low. Sixteen patients were treated with GH and six of 23 without GH therapy were followed for 1 year after GHST. The mean (range) of HV was 7.7 (4.9 to 11.1) cm/yr in patients with GH therapy and 3.7 (2.7 to 4.5) cm/yr in those without GH therapy, which was statistically significant (P<0.001). In the GH-treated group, HV and difference in height SDS during the treatment increased significantly (P<0.001; P< 0.001, respectively). HV increased after 1 year of GH therapy in the SGA and AGA groups (SGA, P=0.043; AGA, P=0.003). The level of Insulin-like growth factor-I was significantly lower in GH-treated patients with height SDS <-3 than those with ${\geq}3$ (P=0.023). Conclusion : In children with growth failure and normal GHST, HV increases significantly by short-term GH therapy. The assessment of long-term effects of GH therapy is necessary. Moreover, further studies should be considered to evaluate the GH-IGF-I axis due to the possibility of GH insensitivity syndrome.

High-level Production of Recombinant Human IFN-$\alpha2a$ with Co-expression of $tRNA^{Arg(AFF/AGA)}$ in High-cell-density Cultures of Escherichia coli

  • Shin, Chul-Soo;Hong, Min-Seon;Shin, Hang-Chel;Lee, Jeewon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.4
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    • pp.301-305
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    • 2001
  • The co-expression of the arg U gene in a double-vector expression system of recombi-nant Escherichia coli BL22(DE3)[pET-IEN2a+pAC-argU] significantly enhanced the production level of reconminant human interferon -$\alpha$2a(rhIFN-$\alpha$2a) in high cell density cultures, compared to a recombinant E. coli culture containing only the single expression vector, pET-IEN2a. The dry cell mass concentration increased to almost 100 g/L, and more than 4 g/L of rhIFN-$\alpha$2a was accumu-lated in the culture broth. Evidently, the synthesis of rhIFN-$\alpha$2a was strongly dependent on the pre-induction growtih rate and more efficient at a higher specific growth rate. The additional sup-ply of tRN $A^{Arg(AGG/AGA)}$ enhanced the expression level of the rhIFN-$\alpha$2a gene in the early stage of the post-induction phase, yet thereafter the specific production rate of rhIFN-$\alpha$2a rapidly de-creased due to severe segregational instability of plasmid vector pET-IEN2a. It would appear that the plasmid instability with only occurred to pET-IEN2a in the double vector system, was re-lated to the effect of translational stress due to the over expression of rhIFN-$\alpha$2a.

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Cell Surface Display of Arylsulfatase Gene from Pseudoalteromonas carageenovora in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 Pseudoalteromonas carageenovora 유래 Arylsulfatase 유전자의 표층 발현)

  • Cho, Eun-Soo;Kim, Hyun-Jin;Jung, So-A;Kim, Jeong-Hwan;Kim, Yeon-Hee;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.355-360
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    • 2009
  • In this study, the arylsulfatase gene (astA, 984 bp ORF) from Pseudoalteromonas carrageenovora genome was expressed on the cell surface of S. cerevisiae by fusing with Aga2p linked to the membrane anchored protein, Aga1p. The constructed plasmid, pCTAST (7.1 kb), was introduced to S. cerevisiae EBY100 cell, and yeast transformants on YPDG plate showed the hydrolyzing activity for 4-methylumbelliferyl-sulfate and p-nitrophenyl-sulfate. When S. cerevisiae EBY100/pCTAST was grown on YPDG medium, the arylsulfatase activity of cell pellet reached about 1.2 unit/mL, whereas no extracellular arylsulfatase activity was detected. The DNA ladder in agarose prepared from agar by this recombinant arylsulfatase showed similar resolution and migration patterns with a commercial agarose. This results revealed that arylsulfatase expressed on the cell surface of S. cerevisiae could be applicable to the economic production of electrophoretic-grade agarose.