• Asian-Australasian Journal of Animal Sciences
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• v.28 no.6
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• pp.788-795
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• 2015

Two dimensional-fluorescence difference gel electrophoresis (2D DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. The purpose of this study was to investigate bovine pregnancy-specific proteins in the proteome between bovine pregnant and non-pregnant serum using DIGE technique. Serums of 2 pregnant Holstein dairy cattle at day 21 after artificial insemination and those of 2 non-pregnant were used in this study. The pre-electrophoretic labeling of pregnant and non-pregnant serum proteins were mixed with Cy3 and Cy5 fluorescent dyes, respectively, and an internal standard was labeled with Cy2. Labeled proteins with Cy2, Cy3, and Cy5 were separated together in a single gel, and then were detected by fluorescence image analyzer. The 2D DIGE method using fluorescence CyDye DIGE flour had higher sensitivity than conventional 2D gel electrophoresis, and showed reproducible results. Approximately 1,500 protein spots were detected by 2D DIGE. Several proteins showed a more than 1.5-fold up and down regulation between non-pregnant and pregnant serum proteins. The differentially expressed proteins were identified by MALDI-TOF mass spectrometer. A total 16 protein spots were detected to regulate differentially in the pregnant serum, among which 7 spots were up-regulated proteins such as conglutinin precursor, modified bovine fibrinogen and IgG1, and 6 spots were down-regulated proteins such as hemoglobin, complement component 3, bovine fibrinogen and IgG2a three spots were not identified. The identified proteins demonstrate that early pregnant bovine serum may have several pregnancy-specific proteins, and these could be a valuable information for the development of pregnancy-diagnostic markers in early pregnancy bovine serum.

• Korean Journal of Veterinary Research
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• v.8 no.2
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• pp.117-120
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• 1968

Normal value of serum proteins of cockerels and pullets was obtained by seperating serum proteins of 80(♂40, ♀40)immature domestic fowls for routine clinical application purpose, using paper and agar electrophoresis. The concentration count of serum proteins which were fractionated was determined by using densitometer and planimeter. The fractionation value of serum proteins (of cockerels and pullets was indicated in the table 1. The composition of serum proteins by paper and agar electrophoresis showed remarkable differences between cockerels and pullets, but only ${\beta}$-globulin could not found difference between them.

• Journal of Life Science
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• v.5 no.2
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• pp.57-62
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• 1995

Casein or soybean protein was subjected to there action with caffeic acidtyrosinase system at 30-35$\circ$C, pH 6.8 with aeration for 5hr. The resulting brown proteins were washed with acetone until the washings were on longer colored. However, modified protein still retained a light brown. The effects of the modified proteins and brown compounds on male Wistar strain rats were studied by pair-feeding of a cholesterol-free diet for 14days. Significant decrease in protein digestibility for the rats fed with the modified proteins were observed. Weight gain and protein digestibility were not influenced by feeding brown compounds, but the feeding of brown compound from casein caused an enlargement of caecum. The concentrations of serum cholesterol and triglyceride in the rats fed with modified proteins and brown compounds were mostly unchanged against the rats fed with untreated proteins. These results suggest that the decrease in protein digestibility induced by enzymic browning-reaction did not cause the decrease in concentration of serum cholesterol.

• Journal of Microbiology
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• v.34 no.1
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• pp.95-100
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• 1996

The effect of the extracellular protease of Salmonella schottmulleri on human serum constituents such as immunoglobulins, hemoglobin and lysozyme and tissue constituents such as fibronectin and collagens was investigated. This protease degraded collagens (type I and III), fibronectin and serum proteins such as human hemoglobin and lysozyme. Bovine serum albumin was degraded slightly. Thus, the present study suggested the possibility that this protease is not only played an important role in invasion of S. schottmulleri by degrading the constituent proteins such as collagens and fibronectin but also induced complications observed in septicemia and chronic infections by degrading the serum proteins. This protease is also capable of degrading defence-oriented humoral proteins, immunoglobulins (IgG and IgM). Furthermore, it is toxic to HEp-2 cells. These findings clarified the possible role of Salmonella protease as a virulence factor in the pathogenesis of Salmonella infections.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.7
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• pp.3265-3270
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• 2012

Clinically, elevated cancer antigen 125 (CA-125) in blood predicts tumor burden in a woman's body, especially in the ovary, but cannot differentiate between malignant or benign. We here used intensive modern proteomic approaches to identify predictive proteins in the serum of women with elevated CA-125 to differentiate malignant from benign ovarian tumors. We identified differentially expressed proteins in serum samples of ovarian cancer (OC) patients, benign ovarian tumor (BT) patients, and healthy control women using mass spectrometry-based quantitative proteomics. Both the OC and BT patients had elevated CA-125. Quantitation was achieved using isobaric tags for relative and absolute quantitation. We obtained 124 quantified differential serum proteins in OC compared with BT. Two proteins, apolipoprotein A-4 (APOA4) and natural resistance-associated macrophage 1, were verified using Western blotting. Proteome profiling applied to OC cases identified several differential serum proteins in the serum of women with elevated CA-125. A novel protein, APOA4, has the potential to be a marker for malignant tumor differentiation in the serum of women with elevated CA-125.

• Animal cells and systems
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• v.15 no.2
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• pp.147-154
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• 2011

Animal cell cultures generally require a nutrient-rich medium supplemented with animal serum. Adult bovine serum contains a variety of nutrients including inorganic minerals, vitamins, salts, proteins and lipids as well as growth factors that promote animal cell growth. To evaluate the potential use of gender-specific bovine serum (GSBS) for cell culture, the biochemical properties of male serum (MS), female serum (FS) and castrated-male serum (CMS) were investigated. Overall, the chemical profile of GSBS was similar to that of bovine references except for glucose, creatine kinase, lactate dehydrogenase and potassium. FS showed elevated total protein and sodium concentrations compared to MS and CMS. Proteins present in MS, FS and CMS but absent in fetal bovine serum (FBS) were selected by two-dimensional gel electrophoresis and identified by peptide mass fingerprinting. Some of the identified proteins are known to be involved in immune responses and the others have unknown physiological roles. Moreover, it was found that some proteins such as alpha-2-macroglobulin appeared to be gender-specific with higher contents in FS. Insulin and testosterone was significantly higher in MS, and $17{\beta}$-estradiol and estrone were higher in FS, as compared to the other sera. Taken together, the results indicate that each GSBS has a different ratio of components. Differences in serum constituents may affect cell cultures in a different manner and could be beneficial, depending on the specific aim of cell cultures.

• Journal of Life Science
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• v.5 no.2
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• pp.1-1
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• 1995

Casein or soybean protein was subjected to there action with caffeic acidtyrosinase system at 30-35$\circ$C, pH 6.8 with aeration for 5hr. The resulting brown proteins were washed with acetone until the washings were on longer colored. However, modified protein still retained a light brown. The effects of the modified proteins and brown compounds on male Wistar strain rats were studied by pair-feeding of a cholesterol-free diet for 14days. Significant decrease in protein digestibility for the rats fed with the modified proteins were observed. Weight gain and protein digestibility were not influenced by feeding brown compounds, but the feeding of brown compound from casein caused an enlargement of caecum. The concentrations of serum cholesterol and triglyceride in the rats fed with modified proteins and brown compounds were mostly unchanged against the rats fed with untreated proteins. These results suggest that the decrease in protein digestibility induced by enzymic browning-reaction did not cause the decrease in concentration of serum cholesterol.

• Journal of Applied Biological Chemistry
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• v.60 no.1
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• pp.55-59
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• 2017

Although wheat is a common staple food in the world, some people suffer from a variety of wheat allergies. For example, wheat-dependent exercise-induced anaphylaxis is induced in the gastrointestinal tract by wheat proteins. Relatively high molecular weight proteins that are salt-insoluble induce many wheat allergies. In the present study, we investigated the induction of an allergy response using crude wheat proteins, which are relatively low molecular weight, salt-soluble proteins. The crude antigen used in this study was extracted using phosphate buffered saline. When the antigen extracts from various wheat cultivars were orally administered, differentiable degrees of allergy responses were observed as measured by serum IgE and histamine secretion compared to the control. Serum IgE levels increased following administration of three of the wheat extracts. This evidence suggests that a combination of salt-soluble wheat proteins could be antigens for the induction of various allergy responses.

• Food Science and Biotechnology
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• v.18 no.5
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• pp.1273-1278
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• 2009

This study aimed to evaluate the potential allergenicity of Cry proteins in insect-resistant genetically modified (GM) maizes (Bt11, MON810, and MON863) using serum screening tests. Serum samples were obtained from Korean children (0-15 years old) with allergic symptoms who had positive maize-specific IgE. The levels of serum specific IgE was measured by the Phadia ImmunoCAP system and considered as positive when they are 0.35 kU/L or higher. Cry proteins (Cry1Ab in Bt11, mCry1Ab in MON810, and Cry3Bb1 in MON863) were expressed in Escherichia coli and purified for serum screening. The reactivity of purified Cry proteins was confirmed by IgE immunoblots in 50 patients (maize-sensitized patients). There was no reaction between Cry proteins and sera from maize-sensitized patients. Our results suggest that these Cry proteins are not likely to cause allergic reactions. Further studies using more sera from patients with true clinical allergies are needed to evaluate the potential allergenicity of novel proteins in GM maize.

• BMB Reports
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• v.46 no.12
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• pp.582-587
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• 2013

Adipocyte differentiation is a complex developmental process forming adipocytes from various precursor cells. The murine 3T3-L1 preadipocyte cell line has been most frequently used in the studies of adipocyte differentiation. Differentiation of 3T3-L1 preadipocytes includes a medium containing fetal bovine serum (FBS) with hormonal induction. In this study, we observed that differentiation medium containing adult bovine serum (ABS) instead of FBS did not support differentiation of preadipocytes. Impaired adipocyte differentiation was due to the presence of a serum protein factor in ABS that suppresses differentiation of preadipocytes. Using a proteomic analysis, alpha-2-macroglobulin and paraoxonase/arylesterase 1, which were previously shown to suppress differentiation of preadipocytes, were identified as anti-adipogenic proteins. Although their functional mechanisms have not yet been elucidated, the anti-adipogenic effects of these proteins are discussed.