• Journal of the Korea Society of Computer and Information
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• v.14 no.7
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• pp.41-47
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• 2009

A solution to the problems of recognizing as one spot or detection failures for complex regions, in which many spots representing proteins are overlapped and saturated, is suggested. The accumulated gradients of each point in complex regions are calculated, and the resulting accumulated gradient image segmented using watershed technique. The suggested solution show better and efficient result than existing method for spot separation, detects more protein spots hidden in the image of 2-dimensional electrophoresis, and expands the scope of prediction.

• Childhood Kidney Diseases
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• v.12 no.2
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• pp.150-156
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• 2008

Purpose : Many results have reported a correlation between the spot urine protein/creatinine ratio(P/C ratio) and 24-hour urinary protein(24UP) amount. This study was designed to evaluated correlation between 24UP amounts and P/C ratio in children and to find the factors that affect this correlation. Methods : 210 patients who visited the Department of Pediatrics in Busan Paik Hospital from september 2003 to december 2007 were included in this study. All the patients were divided into I, II, III/A, B, C group[I:24UP(mg/$m^2$/day)]<100, II: 100$\leq$24UP<1,000, III: 24UP$\geq$1,000, A: Cr excretion(mg/kg)<15, B: 15$\leq$Cr excretion<25, C: Cr excretion$\geq$25)]. Pearson correlation analysis was performed between 24UP and P/C ratio to evaluate the relationship. We defined fractional difference between 24UP and P/C ratio, and then performed multiple regression analysis. Results : There was a strong positive linear correlation between 24UP and P/C ratio in all patients, and the correlation was also good in each group. The factors affecting accurate quantitation of proteinuria using spot urine P/C ratio was creatinine excretion. Conclusion : Spot urine P/C ratio is a useful test to predict proteinuria roughly. Therefore, we expect that urine P/C ratio can be used as parameter instead of 24UP, if we set cutoff value of P/C ratio considered to creatinine excretion according to age and sex in large pediatric population.

• Reproductive and Developmental Biology
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• v.37 no.3
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• pp.149-154
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• 2013

The aim of this study was to evaluate the changes of protein patterns in granulosa cells and corpus luteum in ovaries during the estrus cycle in cows. The estrus cycle was devided into five steps of follicular, ovulatory, early-luteal, mid-luteal and late-luteal phases. In results, 61 spots of total 85 spots were repeated on follicular phase and 51 spots of total 114 spots were repeated on ovulatory phase. The 40 spots of total 129 spots were repeated on early-luteal phase and 49 spots of total 104 spots were repeated on mid-luteal phase. Also 41 spots of total 60 spots were repeated on late-luteal phase. On the other hands, the 16 spots were indicated difference in follicular phase and ovulation phase had a difference 10 spots. It was showed difference No. 103 spot in ovulation phase, No. 135 spot in early-luteal phase and No. 175 and 176 spots in mid-luteal phase. Also, the 11 spots were expressed specifically in mid-luteal phase and No. 178 and 179 spots were difference of expression in late-luteal phase. We confirmed that there were 7 spots for ovulation, 4 spots for luteinization and 2 spots for luteolysis. Spot No. 89~93 in ovulation phase were transferrin, and spot No.94~98 were HSP60. Spot No. 103 was Dusty PK, spot No. 135 was OGDC-E2, and spot No. 175 and 176 were Rab GDI beta from luteinization. Spot No. 178 and 179 in luteolysis were vimentin. This results suggest that will be help to basic data about infertility.

• Clinical and Experimental Pediatrics
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• v.46 no.2
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• pp.173-177
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• 2003

Purpose : Recently, different results about factors affecting accurate quantitation of 24-hr urinary protein(24UP) amount using spot urine protein/creatinine ratio(PCR) have been reported. The current study was designed to evaluate correlation between 24UP amounts and PCR in children, and the effect of 24UP amounts, age, sex, and glomerular filtration rate(GFR) on this correlation. Methods : Among 94 patients who visited the department of pediatrics in Busan Paik Hospital from March 2002 to August 2002, 68 patients whose urinary creatinine excretion was ${\geq}15mg/kg/day$ were included in this study. All the patients were divided into I, II/A, B group(I : 24UP<500 mg/day, II : $24UP{\geq}500mg/day$, A : <10 years of age, B : ${\geq}10years$ of age). Pearson correlation analysis was performed between 24UP and PCR to evaluate the relationship. We defined fractional difference between 24UP and PCR, and then performed multiple regression analysis with 24UP amount, age, GFR and fractional difference. Results : There was a strong positive linear correlation between 24UP and PCR(R=0.936, P<0.0001) in all patients, and the correlation was also good in each group. Using PCR cutoff values of 0.5, the PCR provided high sensitivity, specificity, positive and negative predictive value in predicting 24UP amount ${\geq}500mg$. The factors affecting accurate quantitation of proteinuria using spot urine PCR was age, not 24UP amount, GFR or sex. Conclusion : Spot urine PCR is a useful test but has limitations in predicting 24UP amount. Therefore, it should be used only as screening method. Age-adjusted PCR cutoff values may be necessary to predict 24UP amount in children with proteinuria.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.558-561
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• 2002

In this study, a method for the localization of pregnancy-specific proteins from cow urine on 2-D gel have been established. The proteins were digested with trypsin in gel and then analyzed with MALDI- TOF or transferred to a membrane and microsequenced. To examine the pregnancy associated protein spot 2 as a diagnosis marker in bovine urine 2-D western blotting was performed. This antibody was reacted specifically in the protein of pregnant cow’s urine. Consequently spot 2 was identified and found to be good candidates for developing cow pregnancy detection assay kit.

• Korean Journal of Veterinary Research
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• v.45 no.2
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• pp.155-161
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• 2005

Proteomics is a useful approach to know protein expression, post-translational modification and protein function. We investigated the protein expression pattern and identity in chickens fed with the tissue culture medium waste after harvest of Korean wild ginseng (TCM-KWG) (Panax ginseng). Two groups (n=60/group) of day old broiler chickens were administered with 0 (control) and 0.8% (treatment) TCM-KWG through drinking water. After 5 weeks, we examined the protein expression pattern of fibularis longus and superficial pectoral muscle by Two-dimensional electrophoresis analysis. Interestingly, TCM-KWG treatment significantly increased five spot's density, and markedly reduced five spot's density in the muscles. We identified 10 proteins (desmin, myosin light chain 1, heat shock 25 kDa protein, collapsin response mediator protein-2A, alpha enolase, vimentin, actin alpha 1, my023 protein, pyruvate kinase and troponin T) by the matrix-assisted laser desorption ionization time of flight (MALDI-TOF).

• Reproductive and Developmental Biology
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• v.29 no.3
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• pp.187-191
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• 2005

This study was designed to investigate the changes of quantity and quality of proteins in medium and cytoplasm during in vitro maturation of bovine oocytes. The total quantity of proteins in medium decreased from 0 to 4.5 hr, but increased from 13.5 to 18 hr after the onset of in vitro maturation. The total quantity of protein in cytoplasm increased from 0 to 4.5 hr, decreased from 4.5 to 9 hr, and increased after 18 hr after the onset of in vitro maturation. A total of 298 protein spots was detected on a gel of 2D SDS-PAGE form maturation medium. Among 28 protein spots expressed significant differences in their quantity, 8 proteins were identified by peptide mass fingerprinting (aldose reductase, alpha enolase, apolipoprotein A-1 precursor, 43kDa collectin precursor, heat shock 27kDa protein, plasminogen activator inhibitor-1 precursor, thrombospondin 1, transitional endoplasmic reticulum ATPase). Among total of 35 protein spots detected on gel of 2D SDS-PACE from oorytes cytoplasm, $\beta$-tubulin was identified by peptide mass fingerprinting.

• Journal of Life Science
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• v.17 no.12
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• pp.1729-1733
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• 2007

Brassinolide insensitive associated receptor kinase 1(BAK1) is a critical component that play an important roles in signaling of brassinosteroid biosynthesis. Brassinosteroid-deficient and -insensitive mutants showed the characteristic of dwarf symptom. The nutrient deficient tomato showing stunt phenomenon was selected from soiless cultivation system using modified Sonneveld hydroponic solution. Twenty eight protein spots showing different expression levels compared to the control were isolated from extracts of stunted tomato leaves by 2D PAGE analyses. Significantly down-regulated 6 protein spots out of 28 protein spots were analyzed and sequenced by MALDI-TOF mass spectrometry. The protein spot having pI=4.5 and MW=24 kDa was identified as a signal protein, BAK1, which is directly related to brassinosteroid biosynthesis. In addition, five other protein spots were identified as BCK1, cystein proteinase, sulfutase, peroxidase and zinc finger factor respectively, and they were also signal proteins related to brassinosteroid biosynthesis. Furthermore, amplification of 500bp of BAK1 mRNA by RT-PCR using a primer set of peptide matched regions was inhibited conpared to that of the wild type. The results sugested that the BAK1 might be regulated at the transcription level in response to nutrition applications.

• The Plant Pathology Journal
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• v.19 no.2
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• pp.123-127
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• 2003

A severe disease of muskmelon (Cucumis melo cv. Alsnight) grown on rockwool in a plastic house was characterized by leaf and stem necrosis followed by death of the plants. In 2001, an isolate of Melon necrotic spot virus-MN (MNSV-MN) of the genus Camovirus was identified as the causal agent of the disease on the basis of biological reactions and nucleotide sequence analyses of coat protein (CP) gene. MNSV-MN induced necrotic local lesions on mechanically inoculated leaves and systemic necrotic spots on the upper leaves of melon cvs. Alsnight, Rui III, Party, Imperial, and Seolhang. However, the inoculated leaves of watermelon and cucumber showed only necrotic lesions. DsRNAs extracted from the melon infected with MNSV-MN were separated into three components. Molecular sizes of the dsRNAs were estimated at approximately 4.5, 1.8, and 1.6 kbp. The amplified cDNA products of CP gene for MNSV-MN by RT-PCR showed approximately 1.2 kbp. The amplified DNA was digested to three fragments by MspI treatment. The cDNA of the genomic RNA of MNSV-MN was cloned and the region deduced to encode the CP was sequenced. The CP coding region, located near 3' end of the genome, consisted of 1,170 nucleotides and had the potential to encode a 390 amino acid protein. The nucleotide and amino acid sequences of MNSV-MN CP gene were 84.0-94.6% and 90.8-94.9% identical with other MNSV isolates found in the GeneBank database, respectively. This is the first report on the occurrence of MNSV in Korea.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.32-32
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• 2003

소 난자의 체외성숙 과정에서 세포질 내 단백질의 생산과 축적의 변화는 핵 및 세포질 성숙과 밀접한 관계가 있다는 보고가 있지만, 난자의 성숙과 관련된 특정 단백질의 종류에 대한 보고는 없었다. 따라서 본 연구는 한우 난자의 체외성숙과 관련된 단백질의 생산 및 축적의 변화와 그 종류에 대해서 검토하였다. 체외성숙 시간(4.5, 9, 13.5, 18 및 24시간)에 따른 배양액 내의 단백질 합성의 변화는 2D gel electrophoresis를 이용하였고, 단백질 spot에 대해서는 peptide mass fingerprinting(PMF) 방법을 이용하였다. 또한 단백질 측정 시간에 신선 체외성숙 배양액으로 교환 후 난포란의 핵성숙과 배발달율을 검토하였다. 그 결과 한우 난포란의 체외성숙 시간에 따라 배양액에서 단백질의 양 및 질적인 변화를 확인하였다. 그리고 총 296개 단백질 spot들을 확인하였고, 그 중 30개 spot에서 유의적인 변화가 인정되었다. 또한 유의적인 변화를 보인 spot에 대한 PMF 분석을 통하여 Apolipoprotein A-1 precursor, Alpha enolase, Aldose reductase, 43kDa collectin precursor, Heat shock 27kDa protein, Plasminogen activator inhibitor-1 precursor, Thrombospondin 1 및 Transitional endoplasmic reticulum ATPase가 동정되었다 그리고 총 단백질 합성 경향은 0∼4.5 시간에는 감소하였고, 13.5∼18시간에 증가 한 후 다시 감소하는 경향을 나타내었으며, 단백질의 종류도 시간대별로 현저한 변화가 있었다. 한편 단백질을 측정하는 시기에 신선 체외성숙 배양액으로 교환한 후 난포란의 핵성숙 및 배발달율을 검토한 결과 18시간 체외성숙군에서 9시간째의 교환이 유의적으로 높은 핵성숙을 나타내었으나, 배발달율에서는 유의성이 인정되지 않았다. 그러나 24시간 체외성숙군에서 18시간째의 배양액 교환은 8세포기 및 배반포 발달율이 유의적(P<0.05)으로 높았다. 연구 결과로부터 소 난자의 체외성숙 시간에 따른 단백질 합성 경향의 차이를 확인하였고, 유의적인 변화를 나타낸 8가지의 단백질을 분리할 수 있었으며, 향후 이들 작용기전에 대한 연구가 필요하다고 사료된다.