• 농업과학연구
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• 제45권4호
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• pp.635-643
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• 2018

Bovine milk is widely consumed by humans and is a primary ingredient of dairy foods. Proteomic approaches have the potential to elucidate complex milk proteins and have been used to study milk of various species. Here, we performed a proteomic analysis using 2-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight mass spectrometer (MALDI-TOF MS) to identify whey proteins in bovine milk obtained soon after parturition (bovine early milk). The major casein proteins were removed, and the whey proteins were analyzed with 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE). The whey proteins (2 mg) were separated by pI and molecular weight across pH ranges of 3.0 - 10.0 and 4.0 - 7.0. The 2-DE gels held about 300 to 700 detectable protein spots. We randomly picked 12 and nine spots that were consistently expressed in the pH 3.0 - 10.0 and pH 4.0 - 7.0 ranges, respectively. Following MALDI-TOF MS analysis, the 21 randomly selected proteins included proteins known to be present in bovine milk, such as albumin, lactoferrin, serum albumin precursor, T cell receptor, polymeric immunoglobulin receptor, pancreatic trypsin inhibitor, aldehyde oxidase and microglobulin. These proteins have major functions in immune responses, metabolism and protein binding. In summary, we herein identified both known and novel whey proteins present in bovine early milk, and our sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed their expression pattern.

• Asian-Australasian Journal of Animal Sciences
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• 제16권7호
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• pp.939-945
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• 2003

Seventeen Chinese indigenous pig breeds and three introduced pig breeds had been carried out by means of vertical polyacrylamide gel electrophoresis (PAGE). According to the results, eight serum protein loci were highly polymorphic except Pi-2 and Cp. The polymorphism information content (PIC) of Hpx was the highest (0.5268), while that of Cp was the lowest (0.0257). The population genetic variation index showed that about 84% genetic variation existed in the population, and the rest of 16% distributed between the populations. The genetic variation of Yimeng black pig and Duroc were the highest and the lowest, respectively. The genetic variation of Chinese indigenous pig breeds was much more than that of exotic groups. Genetic distance results showed that Chinese indigenous pig breeds were classified into four groups with the three introduced pig breeds clustered into another group. The results also supported the geographic distribution of Chinese indigenous pig breeds in certain extent.

• 한국수산과학회지
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• 제43권4호
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• pp.307-313
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• 2010

Stanniocalcin 1 (STC1) is a glycoprotein hormone that is important in the maintenance of calcium and phosphate homeostasis in both fish and mammals. STC1 and its paralog STC2 are expressed in multiple tissues in fishes, although the physiological roles of piscine STCs are still unclear compared with those of mammals. In this study, we cloned olive flounder STC1 (ofSTC1) and ofSTC2 cDNAs into pET28a vector and used E. coli Rosetta (DE3) as the host strain for protein expression. Expression experiments were carried out using isopropyl-$\beta$-D-thiogalactoside (IPTG) and nickel affinity chromatography. We could identify the recombinant proteins as single 29.5 kDa (ofSTC1) and 33.2 kDa (ofSTC2) bands in the insoluble fraction on sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE). These results indicate that ofSTC1 and ofSTC2 were expressed as insoluble proteins in E. coli. Furthermore, the injection of ofSTC1 protein into juvenile tilapia resulted in a decrease of the serum calcium level. These results suggest that the purified fish STC1 and STC2 proteins may be used to elucidate the physiological role of STCs in fishes.

• 동아시아식생활학회지
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• 제4권3호
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• pp.59-65
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• 1994

이 실험에서는 소의 유즙에 있는 성장인자를 세포배양을 통해 확인하고 크로마토그래피를 사용하여 분리 정제하였다. 5%의 우유를 포함하는 배지를 FBS(fetal bovine serum)를 포함한 배지와 비교했을때 Africal green monkey kidney cell의 성장 촉진 효과가 비슷하게 나타났으며, 우유의 성장인자를 분리 정제한 결과 FBS배지에서보다 2,000배 이상의 성장 촉진 효과가 나타났다. 이 성장인자는 hydrophobic column(phenyl-sepharose)과 gel-filtration column(sephadex G-100)을 사용하여 분리했으며 그 결과 milk-deriven growth factor는 phenyl-sepharose에서 50% ethylene glycol step에서 용출되므로 hydrophobic protein임이 증명되었고 size exclusion column chromatography로 부터 분자량이 100,000-15,0000 범위의 고분자 물질임이 밝혀졌다. 또한 우유의 성장인자는 매우 다양하며 그중 본 실험으로부터 정제된 물질이 주요 성장인자로 밝혀졌다.

• Parasites, Hosts and Diseases
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• 제48권3호
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• pp.203-211
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• 2010

Advancements in the field of proteomics have provided great opportunities for the development of diagnostic and therapeutic tools against human diseases. In this study, we analyzed haptoglobin and amyloid A protein levels of vivax malaria patients with combinations of depletion of the abundant plasma proteins, 2-dimensional gel electrophoresis (2-DE), image analysis, and mass spectrometry in the plasma between normal healthy donors and vivax malaria patients. The results showed that the expression level of haptoglobin had become significantly lower or undetectable in the plasma of vivax malaria patients due to proteolytic cleavage when compared to healthy donors on 2-DE gels. Meanwhile, serum amyloid A protein was significantly increased in vivax malaria patient's plasma with high statistical values. These 2 proteins are common acute phase reactants and further large scale evaluation with a larger number of patient's will be necessary to establish the possible clinical meaning of the existential changes of these proteins in vivax malaria patients. However, our proteomic analysis suggests the feasible values of some plasma proteins, such as haptoglobin and serum amyloid A, as associating factor candidates for vivax malaria.

• 대한수의학회지
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• 제22권2호
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• pp.115-119
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• 1982

Total protein levels of 60 bovine bull sera were treasured with refractometer and the strum proteins were fractionated by cellulose acetate membrane electorphoresis and the relative amount of each fraction was measured with automatic scanning densitometer. The sixty bovine bulls consisted of 17 Charolais bulls, 28 Holstein bulls and 15 Korean bulls. Mean total serum protein level of the total bulls was $7.50{\pm}0.50(Mean{\pm}SD)$, with the mean of Charolais $7.04{\pm}0.37g/dl$, the mean of Holstein $7.62{\pm}0.40g/dl$ and the mean of Korean $7.81{\pm}0.43g/dl$. As barbital-calcium buffer was used in fractionating the bovine serum protein with cellulose acetate membrane, electric current of 0.4 mA per centimeter width of the membrane for an hour resulted in more clear separation between ${\alpha}_1$-globulin and ${\alpha}_2$-globulin than electric current of 0.6 or 1.0 mA for an hour. However ${\alpha}_1$-globulin and ${\alpha}_2$-globulin could not be measured separately with automatic scanning densitometer. Mean serum albumin level of total bulls was $3.54{\pm}0.42g/dl$, with the mean of Charolais $3.42{\pm}0.26g/dl$, the mean of Holstein $3.69{\pm}0.38g/dl$ and the mean of Korean $3.39{\pm}0.55g/dl$. Mean serum ${\alpha}$-globulin level of the total bulls was $0.62{\pm}0.14g/dl$, with the mean of Charolais $0.61{\pm}0.07g/dl$, the mean of Holstein $0.60{\pm}0.11g/dl$ and the mean of Korean $0.67{\pm}0.23g/dl$. Mean ${\beta}$-globulin level of the total bulls was $0.85{\pm}0.21g/dl$, with the mean of Charolais $0.74{\pm}0.11g/dl$, the mean of Holstein $0.83{\pm}0.14g/dl$ and the mean of Korean $1.02{\pm}0.30g/dl$. Mean serum ${\gamma}$-globulin level of the total bulls was $2.48{\pm}0.45g/dl$ with the mean of Charolais $2.27{\pm}0.37g/dl$, the mean of Holstein $2.48{\pm}0.44g/dl$ and the mean of Korean $2.72{\pm}0.47g/dl$.

• Asian Pacific Journal of Cancer Prevention
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• 제16권14호
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• pp.5779-5785
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• 2015

The present study was designed to investigate cholangiocarcinoma (CCA) antibodies in hamster serum. Hamster CCA cell lines were processed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. A candidate biomarker was confirmed by immunoprecipitation and western blot, and was further analyzed using ELISA and sera from normal control hamsters, hamsters with opisthorchiasis and hamsters with various stages of CCA, as well as from CCA patients and healthy individuals. One candidate marker was identified as $HSP90{\alpha}$, as indicated by a high level of anti-$HSP90{\alpha}$ in hamster CCA sera. It was found that the levels of anti-$HSP90{\alpha}$ were specifically elevated in the sera of hamsters with CCA compared with other groups and progressively increased with the clinical stage. At the cut-off point of 0.4850 on the receiver operating characteristic curve, anti-$HSP90{\alpha}$ could discriminate CCA from healthy control groups with a sensitivity of 76.2%, specificity of 71.4% and total accuracy 75.5%. In the present study, we have shown that anti-$HSP90{\alpha}$ may be a potential useful serum biomarker to discriminate CCA cases from healthy persons.

• 약학회지
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• 제55권3호
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• pp.203-212
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• 2011

Adipocytes have been known to secrete a number of important proteins called adipokines with roles in energy metabolism, reproduction, cardiovascular function and immunity. In this study we have attempted to identify intensively secretory proteins from 3T3-L1 adipocytes. 3T3-L1 preadipocytes were differentiated into mature adipocytes and then the cells were left in serum-free medium. The supernatant was filtrated and dialyzed. Lyophilized secretome was fractionated by two different methods, 1-D SDS PAGE and RP-FPLC. The tryptic peptides from the gel slices and the FPLC fractions were analyzed by nanoLC/ESI-MS/MS. We identified a total of 303 identical proteins from two methods, 251 proteins from 1-D gel and 184 proteins from RP-FPLC. 86 of them were listed as a secretory protein Finally, we identified many known or unknown secreted proteins existed in the low level including adiponectin, angiotensinogen, bone morphogenetic protein-1 (BMP-1), macrophage migration inhibitory factor (MIF), insulin like growth factor-II (IGF-II), interleukin-6 (IL-6), follistatin-related protein-1, minecan, and resistin. The existence of some of secreted proteins has been confirmed in RNA level. This proteomic experiment is useful for the intensive screening of secretory proteins in many kinds of other cells.

• Asian Pacific Journal of Cancer Prevention
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• 제13권12호
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• pp.6501-6504
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• 2012

Background: Multiple myeloma is the most common malignant plasma cell dyscrasia and ranks second among primary haematological malignancies. This study describes the epidemiologic, clinical and pathologic profile of monoclonal gammopathies seen in the University Hospital of the West Indies (UHWI), a tertiary care referral centre. Materials and Method: A retrospective analysis of 85 cases diagnosed at UHWI over the 5-year period 2003-2007 was conducted. The cases were identified from the bone marrow records as well as the computerized database of the Medical Records Department. Clinical presentation, family and personal history and demographic data were retrieved. Haematological and biochemical results were also analyzed. Results: There were 85 patients diagnosed with monoclonal gammopathies. The M:F ratio was 1.2:1 and the mean age was $65.7{\pm}1.3$ years. Eighty percent of the patients had skeletal pain and 40% experienced weight loss. Of the patients experiencing bone pain 56.7% had multiple lytic lesions, 26.7% had pathological fractures and 26.7% had compression fractures. Seventy-four patients (87.1%) had a haemoglobin level <12.0 g/dL with 52.9% having values <8.0 g/dL. Renal impairment was evident at diagnosis in 36.5%. Hypercalcemia was seen in 26.5% and hyperuricemia in 45.9%. Of the 79 patients who had serum protein electrophoresis performed, 77.2% had at least one monoclonal band and of these 24.6% had a monoclonal protein also present on urine protein electrophoresis. Conclusions: The demographic profile in this group of patients is largely similar to other studies in predominantly Caucasian populations; however there was a notable increase in prevalence of severe disease at presentation, with the majority of patients presenting at the most advanced stage. It is probable that these differences reflect socioeconomic factors and not merely inherent ethnic variation in disease biology.

• 한국수정란이식학회지
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• 제23권1호
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• pp.25-30
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• 2008

This study was performed to identify and analyze the specifically expressed plasma proteins during early pregnancy in both pregnant and non-pregnant Hanwoo. Blood samples were collected at 0 (the day of AI), 2, 3, 4, 7, and 11 weeks after AI from pregnant (n=3) and non-pregnant (n=4) Hanwoo, respectively. The hematological parameters were measured. After 2-dimensional electrophoresis using serum, normalized protein spots were selected for the significant expression variation deviated over two fold in its expression level between two groups. Among 17 spots selected, 15 were identified as albumin, IgG1 heavy chain constant region, haptoglobin, ferrochelatase, fibrinogen, hemopexin. 5 spots were expressed only in non-pregnant specific. The spot identification of 1105 and 6106 was decreased after 3 weeks from AI. However, 2/17 spots were still unidentified. Further studies are needed to analyze the function of the proteins associated with early pregnancy.