• Asian-Australasian Journal of Animal Sciences
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• 제21권7호
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• pp.1003-1010
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• 2008

The objective of the present study was to identify differences in the expression levels of liver proteins between healthy and ketotic cows, establish a liver metabolic interrelationship of ketosis and elucidate the metabolic characteristics of the liver during ketosis. Liver samples from 8 healthy multiparous Hostein cows and 8 ketotic cows were pooled by health status and the proteins were separated by two-dimensional-electrophoresis (2D-E). Statistical analysis of gels was performed using PDQuest software 8.0. The differences in the expression levels of liver proteins (p<0.05) between ketotic and healthy cows were identified by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF-TOF) tandem mass spectrometry. Five enzymes/proteins were identified as being differentially expressed in the livers of ketotic cows: expression of 3-hydroxyacyl-CoA dehydrogenase type-2 (HCDH), acetyl-coenzyme A acetyltransferase 2 (ACAT) and elongation factor Tu (EF-Tu) were down-regulated, whereas that of alpha-enolase and creatine kinase were up-regulated. On the basis of this evidence, it could be presumed that the decreased expression of HCDH, which is caused by high concentrations of acetyl-CoA in hepatic cells, in the livers of ketotic cows, implies reduced fatty acid ??oxidation. The resultant high concentrations of acetyl-CoA and acetoacetyl CoA would depress the level of ACAT and generate more ??hydroxybutyric acid; high concentrations of acetyl-CoA would also accelerate the Krebs Cycle and produce more ATP, which is stored as phosphocreatine, as a consequence of increased expression of creatine kinase. The low expression level of elongation factor Tu in the livers of ketotic cows indicates decreased levels of protein synthesis due to the limited availability of amino acids, because the most glucogenic amino acids sustain the glyconeogenesis pathway; thus increasing the level of alpha-enolase. Decreased protein synthesis also promotes the conversion of amino acids to oxaloacetate, which drives the Krebs Cycle under conditions of high levels of acetyl-CoA. It is concluded that the livers of ketotic cows possess high concentrations of acetyl-CoA, which through negative feedback inhibited fatty acid oxidation; show decreased fatty acid oxidation, ketogenesis and protein synthesis; and increased gluconeogenesis and energy production.

• 대한임상검사과학회지
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• 제47권4호
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• pp.286-291
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• 2015

에쿠올은 인간의 건강에 유익한 효과를 나타낸다. 발효된 콩 식품들은 에쿠올을 함유하고 있으며, 많은 미생물들이 에쿠올 생산과정에 참여하는 것으로 밝혀졌다. 본 연구에서는 한국의 전통 발효 식품인 된장에 대해 조사하였다. 먼저 서로 다른 제조자로부터 수집 된 37개의 된장 샘플들을 대상으로 에쿠올의 농도를 측정하기 위해 LC-MS/MS를 시행하였다. 측정 결과 3개의 된장 샘플에서 에쿠올이 검출되었고, 507 ng/100 g의 농도가 가장 높게 나타났다. 에쿠올을 함유한 된장에서 15개의 미생물 종들이 16S rRNA gene sequence analysis와 2개의 MALDI-TOF MS분석법에 의해 분리, 동정되었으며 Bacillus spp, Paenibacillus spp, Tetragenococcus spp, Stapylococcus spp, and Clostridium species들이 가장 우세한 미생물들이었다. 이 연구결과로 한국의 전통 발효식품인 된장에서도 에쿠올이 검출되었음을 확인하였다.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.263-270
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• 2010

Luteal cells produce progesterone that supports pregnancy. Steroidogenesis requires coordination of the anabolic and catabolic pathways of lipid metabolism. In the present study, the corpus luteum (CL) in early pregnancy established from luteal phase and pregnant phase was analyzed. The first study determined progesterone changes in the bovine CL at day 19 (early maternal recognition period) and day 90 in mid-pregnancy and compared them to the CL from day 12 of the estrous cycle. CL alternation was tested using two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF). Comparing CL from luteal phase to those from pregnant phase counterparts, significant changes in expression level were found in 23 proteins. Of these proteins 17 were not expressed in pregnant phase CL but expressed in luteal phase counterpart, whereas, the expression of the other 6 proteins was limited only in pregnant phase CL. Among these proteins, vimentin is considered to be involved in regulation of post-implantation development. In particular, vimentin may be used as marker for CL development during pregnancy because the expression level changed considerably in pregnant phase CL tissue compared with its luteal phase counterpart. Data from 2-DE suggest that protein expression was disorientated in mid pregnancy from luteal phase, but these changes was regulated with progression of pregnancy. These findings demonstrate CL development during mid-pregnancy from luteal phase and suggest that alternations of specific CL protein expression may be involved in maintenance of pregnancy.

• Journal of Plant Biotechnology
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• 제32권4호
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• pp.251-256
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• 2005

Cytokinin은 식물의 성장과 발달에 중요한 역할을 하는 필수 호르몬이다. Cytokinin의 작용 기작을 이해하기 위하여 단백체를 이용하여 cytokinin 관련 단백질들을 동정하였다. 대조구와 t-zeatin을 처리한 애기장대로부터 추출한 단백질을 이차원 전기영동하여 분석하였다. 발현양에 차이가 있는 단백질 spot들을 MALDI-TOF 단백질 질량분석기와 database 검색을 통하여 동정하였다. 그 결과 t-zeatin 처리에 의하여 발현이 증가하는 10개의 단백질과 감소하는 한 개의 단백질을 분리할 수 있었다. Cytokinin에 의하여 발현이 증가하는 단백질은 pollen allergen like protein, L-ascorbate peroxidase, tetrapyrrole methylase family protein, SGT1 protein homolog, disease resistance related protein, maternal embryogenesis control protein, paxneb related protein, gluthathione S-transferase, 그리고 IAA amino acid hydrolase homolog들로 밝혀졌다.

• Reproductive and Developmental Biology
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• 제29권2호
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• pp.83-91
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• 2005

Somatic cell nuclear transfer in cattle has limited efficiency in terms of production of live offspring due to high incidence of fetal failure after embryo transfer to recipients. Such low efficiency of cloning could possibly arise from abnormal and poorly developed placenta. In the present study the placental proteome in late pregnancy established from in vitro fertilization (IVF) and nuclear transfer (NT) was analysed. Proteome alternation was tested using two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI- TOF). Comparing placenta from NT embryos to those from IVF counterparts, significant changes in expression level were found in 18 proteins. Of these proteins 12 were not expressed in NT placenta but expressed in IVF counterpart, whereas the expression of the other 6 proteins was limited only in NT placenta. Among these proteins, cytokeratin 8 and vimentin are considered to be involved in regulation of post-implantation development. In particular, cytokeratin 8 and vimentin may be used as makers for placental development during pregnancy because their expression levels changed considerably in NT placental tissue compared with its IVF counterpart. Data from 2-DE suggest that protein expression was disorientated in late pregnancy from NT, but this distortion was eliminated with progression of pregnancy. These findings demonstrate abnormal placental development during late pregnancy from NT and suggest that alterations of specific placental protein expression may be involved in abnormal function of placenta.

• Journal of Microbiology and Biotechnology
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• 제27권8호
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• pp.1392-1400
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• 2017

Galactooligosaccharides (GOSs) are known to be selectively utilized by Bifidobacterium, which can bring about healthy changes of the composition of intestinal microflora. In this study, ${\beta}-GOS$ were synthesized using bifidobacterial ${\beta}-galactosidase$ (G1) purified from recombinant E. coli with a high GOS yield and with high productivity and enhanced bifidogenic activity. The purified recombinant G1 showed maximum production of ${\beta}-GOSs$ at pH 8.5 and $45^{\circ}C$. A matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of the major peaks of the produced ${\beta}-GOSs$ showed MW of 527 and 689, indicating the synthesis of ${\beta}-GOSs$ at degrees of polymerization (DP) of 3 and DP4, respectively. The trisaccharides were identified as ${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-glucopyranose, and the tetrasaccharides were identified as ${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-galactopyranosyl-($1{\rightarrow}4$)-O-${\beta}-{\text\tiny{D}}$-glucopyranose. The maximal production yield of GOSs was as high as 25.3% (w/v) using purified recombinant ${\beta}-galactosidase$ and 36% (w/v) of lactose as a substrate at pH 8.5 and $45^{\circ}C$. After 140 min of the reaction under this condition, 268.3 g/l of GOSs was obtained. With regard to the prebiotic effect, all of the tested Bifidobacterium except for B. breve grew well in BHI medium containing ${\beta}-GOS$ as a sole carbon source, whereas lactobacilli and Streptococcus thermophilus scarcely grew in the same medium. Only Bacteroides fragilis, Clostridium ramosum, and Enterobacter cloacae among the 17 pathogens tested grew in BHI medium containing ${\beta}-GOS$ as a sole carbon source; the remaining pathogens did not grow in the same medium. Consequently, the ${\beta}-GOS$ are expected to contribute to the beneficial change of intestinal microbial flora.

• 농업과학연구
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• 제42권1호
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• pp.29-35
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• 2015

This study was performed to analyze the distribution and antimicrobial resistance of pathogenic microorganisms isolated from the carcass and environments of chicken processing plant located in Gyeonggi province from October to November in 2010. Chicken slaughterhouse was visited 3 times and totally 40 samples were collected from chicken carcass before and after washing (n=14), chicken cuts (n=7), cooling water (n=8), brine (n=2), cutting knives (n=7) and working plate (n=2). Whole-chicken rinsing technique (for chicken carcasses) and swab technique (for working plate and knives) were used to analyze the distribution of pathogenic microorganisms. In addition, brine and chilling water from storage tanks were gathered using sterilized tubes and used as samples. The matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) for whole cell fingerprinting in combination with a dedicated bioinformatic software tool was used to identify the isolated microorganisms. The pathogenic microorganisms, such as Bacillus cereus (n=8) and Staphylococcus aureus (n=9), were isolated form the chicken processing process (chicken carcasses of before and after chilling, chicken cuts, and working plate). The antimicrobial susceptibility of those isolated microorganisms was analyzed using 21 antimicrobial agents. In the case of B. cereus, it showed 100% of resistance to subclasses of penicillins and peptides, and it also resistant to cephalothin, a member of critically important antimicrobials (CIA), however there was no resistance (100% susceptible) to vancomycin and chloramphenicol. S. aureus showed 100% resistance to subclasses of peptides and some of penicillins (penicillin and oxacillin), however, it showed 100% susceptibility to cephalosporins (cefazolin and cephalothin). All of the tested pathogens showed multi drug resistance (MDR) more than 4 subclasses and one of B. cereus and S. aureus showed resistance to 9 subclasses. After the ban on using the antimicrobials in animal feed in July 2011, there would be some change in microbial distribution and antimicrobial resistance, and it still has a need to be analyzed.

• Molecular & Cellular Toxicology
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• 제1권2호
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• pp.116-123
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• 2005

The application of high pressure on cellular morphology, proliferation and protein expression of Jurkat cells (human T lymphocyte cell line) has been extensively investigated. In the present study, we manufactured a novel pressure chamber that modulates 5% $CO_{2}$, temperature and pressure (up to 3 ATA). Jurkat cells was incubated 2 ATA pressure and analyzed cellular morphology and growth using an electron microscopy and MTT assay. The cells showed the morphological changes in the cell surface, which appeared to cause a severe damage in cell membrane. The growth rate of the cells under 2 ATA pressure decreased as cultured time got increased. Furthermore, a long term exposure of high pressure on Jurkat cells may act as one of the important cellular stresses that leads to inducing cell death. Cellular proteomes were separated by 2-dimensional electrophoresis with pH 3-10 ranges of IPG Dry strips. And many proteins showed significant up-and-down expressions with hyperbaric pressure. Out of all, 10 spots were identified significantly using matrix-assisted laser desorption/ionization-time of fight (MALDI-TOF) mass spectrometry. We and found that 9 protein expressions were decreased and one protein, heat shock protein HSP 60, was increased in Jurkat cells under 2 ATA. Identified proteins were related to lipid metabolism and signal transduction.

• 한국가금학회:학술대회논문집
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• 한국가금학회 2005년도 제22차 정기총회 및 학술발표회
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• pp.66-67
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• 2005

배자 생식세포 발달에 관련된 메카니즘을 밝혀내기 위해서, 닭 배자 생식기에서 추출한 원시 생식세포의 단백질체 지도를 만들었다. 총 500 배자를 6일간 배양하여 배자 생식기를 획득했고, 7-10일 배양 후, 배양된 원시생식세포는 2차원 젤 전기 영동법에 의해 분할되어 졌다. 유의적 발현 수준을 나타낸 많은 단백질 스팟 들은 MALDI-TOP 와 LC-MS/MS에 의해 확인되었으며, 89개의 단백질 스팟 중에 50개의 mass spectra 들이 데이터베이스에서 조류 단백질과 일치함을 확인하였다. 본 실험에서 행한 단백질체 지도는 형질전환 연구와 생식세포 생물학 분야에 중요한 참고 문헌으로 가치를 가질수 있을 것이다.

• 공업화학
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• 제28권3호
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• pp.263-271
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• 2017

최근에, 질량분석기술의 폴리머 분석에의 응용은 MALDI-TOF MS 개발 이후 급속도로 발전하였다. 이 리뷰 논문은 현재까지 연구된 MALDI-TOF MS의 폴리머 특성분석에의 응용에 관한 최신 논문을 정리하였다. MALDI-TOF MS는 바이오 폴리머와, 합성 폴리머의 평균분자량 분석, 폴리머의 시퀀스 분석을 통한 구조의 해석, 모노머의 조성분석에까지 이용되고 있다. 엔드그룹의 특성과 농도를 분석하는 연구도 많이 진행되었고, 복잡한 폴리머의 분자량의 분석에는 SEC와 MALDI-TOF MS를 연결한 분석법을 추천한다. MALDI에 tandem MS를 결합한 분석기술이나, 이온 모빌리티를 응용한 질량분석기, TOF-SIMS, MALDI-TOF-Imaging 기술도 급격히 발전하고 있으며, 이의 폴리머 특성분석에의 응용은 별도의 분리기술이 필요 없어 앞으로 더 많이 이용될 것으로 생각된다. 분자량, 시퀀스, 그리고 모노머의 조성을 정확하게 계산해주는 소프트웨어와 고분자량(> 100 kDa)의 분석을 가능하게 해주는 기술이 개발된다면, 폴리머를 연구하는 과학자들에게 MALDI-TOF MS의 이용은 문제점을 해결하고, 목적하는 폴리머를 합성하는 데 중요한 수단이 될 것이다.