• Title/Summary/Keyword: NS3 protein

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Molecular Characterization of an H5N3 Influenza Virus Isolated from Spot-Billed Duck

  • Lee, Jin Hwa;Kwon, Hyuk Moo;Sung, Haan Woo
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.243-252
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    • 2013
  • Among the 16 hemagglutinin (HA) subtypes of avian influenza virus (AIV), only the H5 and H7 subtypes have caused highly pathogenic avian influenza (HPAI) in poultry. However, most H5 or H7 subtype viruses are categorized as low pathogenic avian influenza (LPAI). Some AIVs, including the H5 and H7 HPAI viruses, have shown the ability to infect humans directly. In this study, we describe the biological and molecular characterization of an H5N3 AIV (SBD/KR/KNU SYG06/06) isolated from spot-billed duck (Anas poecilorhyncha) in Korea. A phylogenetic analysis of the eight viral genes showed that the SBD/KR/KNU SYG06/06 isolate belongs to the Eurasian lineage and that the SBD/KR/KNU SYG06/06 isolate was clearly different from HPAI H5N1 strains, including human isolates and the Italian HPAI H5N2 strains. Additionally, no relationship was found between SBD/KR/KNU SYG06/06 and the Korean HPAI H5N1 isolates. The SBD/KR/ KNU SYG06/06 isolate had avian specific receptor binding site residues in the HA protein and the four C-terminal amino acids in the NS1 protein. The HA protein of the SBD/KR/KNU SYG06/06 isolate exhibited the typical LPAI motif at the cleavage site and this virus produced no cytopathic effects in MDCK cells without trypsin. Given these results, we suggest that the H5N3 AIV isolated from the spot-billed duck should be considered an LPAI virus and should have no pathogenic effect in humans.

BcI-2 Over-expression Reduced the Serum Dependency and Improved the Nutrient Metabolism in a NS0 Cells Culture

  • Tey Beng Ti;Al-Rubeai Mohamed
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.254-261
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    • 2005
  • The over-expression of Bcl-2 has greatly improved the culture period, specific growth rate, and maximum viable cell density of NS0 cells culture under low serum condition. Further analysis of these data suggests that a saturation model of the Monod type can be used to represent the relationships of specific growth rate and initial serum concentration. The ${\mu}_{max}$ and $K_s$ for the Bcl-2 cell line is $0.927day^{-1}\;and\;0.947\%(v/v)$ respectively, which are $21\%$ greate and $7\%$ lower respectively than its control counterpart. Study on the amino acid supplementation revealed that Bcl-2 cell lines possess greater improvement in the specific growth rate and maximum viable cell density compared to the control cell lines. A further increase in the amino acid supplementation has resulted a $17\%$ decrease in specific growth rate and no improvement in maximum viable cell density in the control culture. However, the Bcl-2 cell line exhibited a better growth characteristic in this culture condition compared to that of control cell lines. The higher specific growth rate and maximum viable cell density of the Bcl-2 cell line in medium fortified with serum and MEM EM suggested a more efficient nutrient metabolism compared to that in the control cell line. The low serum and amino acid utilisation rate and the higher cell yield may prove to be important in the development of serum/protein free culture.

Generation of a Mammalian Gene Expression Vector Using Bovine Viral Diarrhea Virus (Bovine Vira1 Diarrhea Virus를 이용한 포유동물세포 발현벡터의 개발)

  • 이영민
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.86-95
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    • 2002
  • As a result of genome projects, the research to elucidate the function of a protein of interest has recently been well-recognized. In order to facilitate functional genomics, a useful mammalian gene expression vector is required. Using an infectious CDNA clone of BVDV pNADLclns-, we have developed a mammalian gene expression vector. In this study, a replication-competent full-length infectious CDNA clone containing puremycin acetyltransferase (pac) gene (pNADLclns-/pac) was successfully generated. The viral RNA replication and viral protein NS3 synthesis were examined by detecting metabollically $^{32}P$-labelled genomic viral RNA and immunoblotting with a mouse anti-NS3 antibody. To generate viral replicon as an expression vector, we examine if the viral structural genes (C, E0, El, E2) are required for viral replication by deletion analysis. As a result, all of the structural proteins are dispensable for viral replication per se, but essential for infectious viral particle formation. Based on our deletion analysis, we have generated a replication-competent BVDV viral replicon (pNADLclns-/pac/${\Delta}S$), whose structural genes are all deleted. In addition to NADLclns- /pac/${\Delta}S$, NADLclns-/ luc/${\Delta}S$ viral replicon containing luciferase gene as a reporter was constructed and fecund to be replication-compotent in HeLa and BHK cells as well as MDBK cells. Therefore, BVDV viral replicon developed in our study will be a useful tool to express a protein of interest in various mammalian cells.

Comparison of Milling and Flour Quality Characteristics of Foreign Wheat and Korean Wheat

  • Jinhee Park;Kyeong-Hoon Kim;Chon-Sik Kang;Go Eun Lee;Kyeong-Min Kim;Mina Kim;Han-yong Jeong;Yurim Kim;Jiyoung Shon;Jong-Min Ko
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.296-296
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    • 2022
  • This study was investigated to compare the milling and physicochemical characteristics of six Korean wheat cultivars (Keumkang, KK; Jokyung, JK; Goso, GS; Joongmo2008, JM; Baekkang, BK; Saekeumkang, SKK) and five foreign wheat classes (Australian standard white wheat, ASW; Australian hard, AH; US northern spring, NS; US hard red winter, HRW; Soft wheat, SW). Korea and foreign wheat grains were milled using a Buhler MLU-202. Flour moisture, ash, protein, gluten, sedimentation, particle size, solvent retention capacity (SRC) and dough properties of flour were analyzed. Results showed that the hard wheats had a greater total flour yield and reduction fraction yield than the soft wheats regardless of the country. However, there were in the milling characteristics between the US and Korean soft wheats. GS, a soft wheat in Korea, had the lowest flour yield (59.6%) and the highest bran fraction yield (21.4%). The particle sizes of flour by milling fraction were B1>B2>B3 for the largest, and the R1〈R2〈R3 for the smallest. Particle size, ash, protein contents and the values of lactic acid SRC showed highly correlated with flour yield. The gluten-performance-index (GPI) is the ratio of the lactic acid SRC value to the sum of sodium carbonate and sucrose SRC values, and it has been used as a quality indicator for overall performance potential of flour. GPI values differed depending on the wheat variety or class, JM (0.82) was the highest value, and SKK (0.56) and SW (0.59) were low. The curve pattern of the Mixolab result also gives a quality indication of the flour sample. JM and NS flour had similar pattern at water absorption and gluten strength parameters and BK and HRW had similar viscosity patterns. These results will enable further study for blending Korean wheat cultivar to improve the flour quality.

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Characterization of Mouse Hybridoma Producing Monoclonal Antibody Paragonimus westermani (폐디스토마에 대한 Mmonoclonal Antibody생산 세포주개발에 대한 연구)

  • 고광삼;이숙영;이근배
    • Korean Journal of Microbiology
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    • v.23 no.1
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    • pp.64-68
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    • 1985
  • In this study, hybridoma techniques were applied to produce monoclonal antobodies to Paragonimus westermani, commonly known as lung distoma. Balb/c mice were immunized intraperitoneally every week with increasing doses of purfied protein of Paragonimus westermani adult worms begining with 0.3/mg/mouse/7 days and ending with 0.5mg at 28th day. Spleen cells from these immunized mice were hybridized with myeloma cells (NS-1) and the hybridized cells were selected in HAT media. The antibody secreting cells among the hybrid cells were initially selected by ELISA. Those initially selected cells were further screened by the criteria of antibody producing activity, and seneral cell lines among them were further tested with immunodiffusion method. One hybridoma clone produced IgM and another clone produced IgG1. The supernatant of the hybridoma clone producing IgM had titer 1:64 and the hybridoma clone producing IgG1 had titer 1:256 measured by immunofluorescence technique.

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Diversity in Lipid Contents and Fatty Acid Composition of Soybean Seeds Cultivated in Korea (국내 유통 콩의 지방함량 및 지방산 조성변이)

  • Kim, Sun-Lim;Lee, Yeong-Ho;Chi, Hee-Youn;Lee, Sun-Joo;Kim, Si-Ju
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.3
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    • pp.348-357
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    • 2007
  • The 117 soybeans seeds were collected from the nine provinces of Korea, and protein and lipid contents, and fatty acid composition levels were evaluated to investigate their relationship. The 100-seed weights of the black soybeans were varied $27.7{\sim}33.1g$, while the 100-seed weight of yellow soybeans were varied $24.6{\sim}36.6g$. Protein and lipid content of the 117 soybean seeds was 38.3% and 17.8%, respectively Protein contents of the 59 black soybean seeds (38.6%) were significantly higher than those of the 58 yellow soybean seeds (37.9%). However, lipid contents of the black soybean seeds (17.6%) were lower than those of the yellow soybean seeds (18.1%). Linoleic and oleic acid composition levels of the 117 soybean seeds were 53.75% and 22.08%. Unsaturated fatty acid levels of soybean seeds showed a statistically significant variability among the nine provinces of Korea, however, the differences were not found in the linoleic (18:2) and oleic acid (18:1) levels. Therefore, it was considered that the significant variability of unsaturated fatty acid were mainly due to the variations of linolenic acid (18:3) level. The composition levels of linoleic, oleic, palmitic (16:0), and linolenic acid in the yellow soybean seeds were 53.43%, 22.73%, 12.23%, and 8.24%, while those of the black soybean seeds were 54.13%, 21.48%, 12.47%, and 8.31%. Obtained results suggested that fatty acid composition levels were varied and possibly influence by the phenotype of seed coat colors. Oleic acid, mono-saturated fatty acid, showed the most remarkable variability between yellow and black soybean seeds, and the composition levels were higher in the yellow soybean seeds. Relationship between unsaturated fatty acid levels and 100-seed weights in the yellow soybean seeds showed a negative correlation (r=-0.513, P<0.01), but no relationship $(r=0.154^{ns})$ was observed in the black soybean seeds.

Clinical Features of Atypical Kawasaki Disease (비정형 가와사키병의 임상 양상)

  • Heo, Mi Young;Choi, Su Jung;Kim, Hae Soon;Sohn, Sejung
    • Clinical and Experimental Pediatrics
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    • v.45 no.3
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    • pp.376-382
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    • 2002
  • Purpose : To identify clinical and laboratory features of atypical Kawasaki disease(KD), and to develop criteria for early diagnosis of atypical KD patients. Methods : All patients with KD treated at our hospital from January 1998 to June 2000 were reviewed retrospectively. Results : Among a total of 167 patients, 28(16.8%) were atypical KD of which seven(25%) were infants. Among the five cardinal symptoms, oral mucosal change(96.4%) occurred most frequently, followed by conjuntivitis(57.1%) and rash(46.4%). Most notable laboratory findings were anemia, and increased erythrocyte sedementation rate(ESR) or C-reactive protein(CRP). Coronary artery abnormalities developed in seven(25.8%) atypical KD patients, compared with 14.4% in typical KD patients. We considered oral mucosal change as major criterion, and conjunctivitis, rash, hematocrit <35% and ESR >30 mm/hr or CRP >3.1 mg/dL as minor criteria. Proposed modification in diagnostic criteria for atypical KD include fever of ${\geq}5$ days+major criterion+${\geq}2$ minor criteria, or fever of ${\geq}5$ days+4 minor criteria. Conclusion : The modified diagnostic criteria has yielded a sensitivity 89.3%. Our diagnostic criteria may be used for early diagnosis of atypical KD.

The Effect of Collagen Peptide Intake on UVB-induced Skin Damage in Hairless mice (콜라겐 펩타이드 섭취 무모생쥐에서 UVB 조사에 의한 피부 손상에 미치는 효과 규명)

  • Shin, Hyun-Jun;Park, Jong il;Kwon, Chang-Ju;Kim, Hwa young;Inoue, Naoki;Koizumi, Seiko;Hwang, Jae Sung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.17 no.3
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    • pp.611-621
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    • 2016
  • This study examined the effects of collagen peptide intake extracted from the tilapia fish (TFCP) on UVB-induced skin in hairless mice. To cause photoaging, the hairless mice were exposed to UVB irradiation three times a week and TFCP was administered with 545 and 1090 mg/kg once a day for 12 weeks. The level of wrinkle formation, thickness of the skin, acute erythema, skin hydration, amount of hydroxyproline, expression of MMPs and filaggrin were measured. Wrinkle formation, thickness of the skin, and acute erythema were significantly lower and skin hydration was higher in the TFCP intake group compared to the UVB-control mice group. In addition, the amount of the collagen and hydroxyproline of the collagen peptide intake group were much higher than the UVB-control group. Oral administration of TFCP inhibited the expression of the MMP-3 and MMP-13 mRNA level as well as activated MMP-2 and MMP-9 induced by UV irradiation, and increased the protein expression of filaggrin in hairless mice skin. In summary, the oral administration of TFCP can inhibit wrinkle formation and protect the skin from UV-damage. Therefore, this study suggests that TFCP is a potential candidate material for skin functional foods.

Preventive Effect on Development of Diabetes and Renoprotective Effect of Acanthopanax Senticosus Aqua-acupuncture in Multiple Low-dose Streptozotocin-Induced Diabetic Rats (가시오가피약침(五加皮藥鍼)이 당뇨유발억제(糖尿誘發抑制) 및 신장보호활성(腎臟保護活性)에 미치는 영향)

  • Chung, Chong-un;Lee, Yun-ho;Kang, Sung-keel
    • Journal of Acupuncture Research
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    • v.20 no.3
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    • pp.1-14
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    • 2003
  • Objective : The aim of the study was to investigate the preventive effect of Acanthopanax senticosus(AS) aqua-acupuncture into Sinsu(BL23) of the multiple low-does strepozotocin(STZ)-induced diabetic rats. Methods : The experimental animals were divided into 4 groups : normal group of rats, control group of multiple low-does STZ-induced diabetic rats, NSAA group with 0.4ml normal saline(NS) aqua-acupunctured subcutaneously into Sinsu in multiple low-does STZ-induced diabetic rats, and ASAA group with 0.4ml of 20% AS aqua-acupunctured subcutaneously into Sinsu in multiple low-does STZ-induced diabetic rats. Each of AS and NS aqua-acupuncture was done subcutaneously into both loci of Sinsu taking turns everyday for 3 weeks. Thereafter the levels of serum glucose, body weight, index of kidney hypertrophy, urine glucose, urinary albumin excretion, creatinine clearance, mesangial cell and TGF-${\beta}1$ expression in glomeruli and tubular cells were measured on the determined day. Conclusions : 1. Both ASAA and NSAA groups decreased the serum glucose levels in multiple low-dose STZ-induced diabetic rats as compared to the cintrol group, and ASAA group showed more significant decreases than NSAA group. 2. Both ASAA and NSAA groups prevented the development of diabetes in multiple low-dose STZ-induced diabetic rats as compared to the control group, and ASAA group prevented more markedly the development of diabetes than NSAA group. 3. Both ASAA and NSAA groups prevented the reduction of body weight in multiple low-dose STZ-induced diabetic rats as compared to the control group, and ASAA group showed the same as the normal group. 4. Both ASAA and NSAA groups did not show any changes of the creatinine clearance in multiple low-does STZ-induced diabetic rats. 5. Both ASAA and NSAA groups prevented the excretion of urinary glucose and albumin in multiple low-dose STZ-induced diabetic rats as compared to the cintrol group, and ASAA group showed more significant prevention than NSAA group. 6. Both ASAA and NSAA groups prevented the expansion of glomerular cells and the protein expression of transforming growth factor-${\beta}1$ in multiple low-dose STZ-induced diabetic rats as compared to the cintrol group, and ASAA group prevented more significantly than NSAA group.

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Efficacy and Safety of Deflazacort in Korean Children with Nephrotic Syndrome (신증후군 환아에 대한 Deflazacort의 효과와 안전성)

  • Kim, Mi Jeong;Jeon, Kye Won;Jin, Dong Kyu;Lee, Suk Hyang
    • Korean Journal of Clinical Pharmacy
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    • v.10 no.2
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    • pp.51-56
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    • 2000
  • Deflazacort, an oxazoline derivative of prednisolone, has been claimed to have anti-inflammatory effects with fewer side effects compared to prednisone. The objectives of the study were to evaluate efficacy and safety of deflazacort in children with nephrotic syndrome. Eligible Patients were the children with nephrotic syndrome who were treated with deflazacort from October. 1994 to April. 1999. Nephrotic syndrome was defined as having albumin level of less than 2.5 mg/dL and 24-hour urinary protein excretion of greater than $40\;mg/m^2/hr$. The primary parameters evaluating the efficacy of deflazacort were response rate, time to respond and relapse frequency. The safety profiles were the impact on children's growth, calcium sparing effect, glucose metabolism, lipid profile and adverse drug reactions. As results, total of 60 children were evaluated (47 boys, 13 girls). Response rate was $95\%$ (57/60) for initial and late responders. Median time to respond was 12 days (range 7-110 days) and median relapse frequency was one time (range 0-6). Weight/height ratio increased from $22.05\pm3.47\;to\23.20\pm3.44\;kg/m$ (p<0.001) and plasma calcium level, from $7.55\pm3.86\;to\;9.98\pm3.77\;mg/dL$ after treatment (p<0.001). Change of fasting glucose level was not statistically significant $(91.92\pm3.53\;vs.\;98.19\pm4.78\;mg/dL,\;p=0.072)$, while change of total cholesterol was significant $(362.3\pm12.0\;vs\;251.4\pm11.5\;mg/dL$, p<0.001). In conclusion, patients on deflazacort showed similar efficacy in treatment of nephrotic syndrome as reported for prednisone with less impact on growth inhibition and metabolic side effects of hyperglycemia and hyperlipidemia.

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