• Asian-Australasian Journal of Animal Sciences
• /
• 제25권11호
• /
• pp.1540-1545
• /
• 2012

Steroidogenesis requires coordination of the anabolic and catabolic pathways of lipid metabolism, but the profile of proteins associated with progesterone synthesis in cyclic and pregnant corpus luteum (CL) is not well-known in cattle. In Experiment 1, plasma progesterone level was monitored in cyclic cows (n = 5) and pregnant cows (n = 6; until d-90). A significant decline in the plasma progesterone level occurred at d-19 of cyclic cows. Progesterone level in abbatoir-derived luteal tissues was also determined at d 1 to 5, 6 to 13 and 14 to 20 of cyclic cows, and d-60 and -90 of pregnant cows (n = 5 each). Progesterone level in d-60 CL was not different from those in d 6 to 13 CL and d-90 CL, although the difference between d 6 to 13 and d-90 was significant. In Experiment 2, protein expression pattern in CL at d-90 (n = 4) was compared with that in CL of cyclic cows at d 6 to 13 (n = 5). Significant changes in the level of protein expression were detected in 32 protein spots by two-dimensional polyacrylamide gel electrophoresis (2-DE), and 23 of them were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Six proteins were found only in pregnant CL, while the other 17 proteins were found only in cyclic CL. Among the above 6 proteins, vimentin which is involved in the regulation of post-implantation development was included. Thus, the protein expression pattern in CL was disorientated from cyclic luteal phase to mid pregnancy, and alterations in specific CL protein expression may contribute to the maintenance of pregnancy in Korean native cows.

• Asian-Australasian Journal of Animal Sciences
• /
• 제31권9호
• /
• pp.1401-1406
• /
• 2018

Objective: The uncoupling protein 3 (UCP3) is a member of the mitochondrial anion carrier superfamily and has crucial effects on growth and feed efficiency in many species. Therefore, the objective of the present study was to examine the association of polymorphisms in the UCP3 gene with feed efficiency in meat-type chickens. Methods: Six single nucleotide polymorphisms (SNPs) of the UCP3 gene were chosen to be genotyped using matrix-assisted laser desorption-ionization time-of-flight mass spectrometry in meat-type chicken populations with 724 birds in total. Body weight at 49 (BW49) and 70 days of age (BW70) and feed intake (FI) in the interval were collected, then body weight gain (BWG) and feed conversion ratio (FCR) were calculated individually. Results: One SNP with a low minor allele frequency (<1%) was removed by quality control and data filtering. The results showed that rs13997809 of UCP3 was significantly associated with BWG and FCR (p<0.05), and that rs13997811 had significant effects on BW70 and BWG (p<0.05). Rs13997812 of UCP3 was strongly associated with BW70, FI, and FCR (p<0.05). Furthermore, individuals with AA genotype of rs13997809 had significantly higher BWG and lower FCR (p<0.05) than those with AT genotype. The GG individuals showed strongly higher BW70 and BWG than AA birds in rs13997811 (p<0.05). Birds with the TT genotype of rs13997812 had significantly greater BW70 and lower FCR compared with the CT birds (p<0.05). In addition, the TAC haplotype based on rs13997809, rs13997811, and rs13997812 showed significant effects on BW70, FI, and FCR (p<0.05). Conclusion: Our results therefore demonstrate important roles for UCP3 polymorphisms in growth and feed efficiency that might be used in meat-type chicken breeding programs.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권7호
• /
• pp.3311-3317
• /
• 2014

Background: The consequence of Rho GDP dissociation inhibitor alpha (RhoGDI${\alpha}$) activity on migration and invasion of estrogen receptor positive ($ER^+$) and negative ($ER^-$) breast cancer cells has not been studied using the proteomic approach. Changes in expression of RhoGDI${\alpha}$ and other proteins interacting directly or indirectly with RhoGDI${\alpha}$ in MCF7 and MDA-MB-231, with different metastatic potentials is of particular interest. Materials and Methods: $ER^+$ MCF7 and ER- MDA-MB-231 cell lines were subjected to two-dimensional electrophoresis (2-DE) and spots of interest were identified by matrix-assisted laser desorption/ionization time of- flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry (MS) analysis after downregulation of RhoGDI${\alpha}$ using short interfering RNA (siRNA) and upregulated using GFP-tagged ORF clone of RhoGDI${\alpha}$. Results: The results showed a total of 35 proteins that were either up- or down-regulated in these cells. Here we identifed 9 and 15 proteins differentially expressed with silencing of RhoGDI${\alpha}$ in MCF-7 and the MDA-MB-231 cells, respectively. In addition, 10 proteins were differentially expressed in the upregulation of RhoGDI${\alpha}$ in MCF7, while only one protein was identified in the upregulation of RhoGDI${\alpha}$ in MDA-MB-231. Based on the biological functions of these proteins, the results revealed that proteins involved in cell migration are more strongly altered with RhoGDI-${\alpha}$ activity. Although several of these proteins have been previously indicated in tumorigenesis and invasiveness of breast cancer cells, some ohave not been previously reported to be involved in breast cancer migration. Hence, these proteins may serve as useful candidate biomarkers for tumorigenesis and invasiveness of breast cancer cells. Conclusions: Future studies are needed to determine the mechanisms by which these proteins regulate cell migration. The combination of RhoGDI${\alpha}$ with other potential biomarkers may be a more promising approach in the inhibition of breast cancer cell migration.

• Bulletin of the Korean Chemical Society
• /
• 제34권8호
• /
• pp.2419-2424
• /
• 2013

Guanosine-5'-diphosphate 3'-diphosphate (ppGpp) serves as alarmone in bacterial stringent responses. In this study, an affinity column was constructed by immobilizing ppGpp to NHS-Sepharose for isolating ppGpp-binding proteins. A novel ppGpp-binding protein, YjgA, was isolated and characterized by MALDI-TOF MS (matrix-assisted laser desorption ionization-time-of-flight mass spectrometry) coupled with two-dimensional gel electrophoresis. YjgA and truncated forms of YjgA were cloned and over-expressed in BL21 (DE3). The binding affinity of YjgA to ppGpp was determined by equilibrium dialysis. The interaction of YjgA with ppGpp was very specific, considering that the dissociation constant of YjgA with ppGpp was measured as $5.2{\pm}2.0{\mu}M$, while the affinities to GTP and GDP were about 60 and 30 times weaker than ppGpp. Expression of yjgA gene in Escherichia coli K-12 MG1655 was examined by reverse transcription polymerase chain reaction (RT-PCR). RT-PCR results revealed that yjgA was expressed from early to late stationary phase. The yjgA deletion mutant exhibited decreased cell number at stationary phase compared to parent strain and the over-expression of YjgA increased the cell number. These results suggested that YjgA might stimulate cell division under stationary phase. In most prokaryotic genome, about half of the protein candidates are hypothetical, that are expected to be expressed but there is no experimental report on their functions. The approach utilized in this study may serve as an effective mean to probe the functions of hypothetical proteins.

• Radiation Oncology Journal
• /
• 제22권4호
• /
• pp.298-306
• /
• 2004

목적:. 방사선 감수성이 다른 마우스 조직에서 apoptosis 유도 수준을 확인하고 방사선 감수성에 관여 된 인자를 Proteomics를 통해서 확인한다. 대상 및 방법: C3H/HeJ 마우스에 10 Gy 방사선을 조사하고 8시간 후 비장과 간을 채취하여 apoptosis 유도 수준을 비교 분석하였다. 조직에서 단백질을 추출하여 2-dimension electrophoresis (2-DE)를 실시하였다. 2-DE에서 방사선에 의해 발현의 변화를 보이는 gel의 spot를 trypsin 처리하여 MALDI-TOF 측정한 후 Swiss-prot database를 통하여 단백질 을 동정하였다. 결과: Apoptosls index는 방사선 조사 후 비장 조직에서 $35.3{\pm}1.7{\%}$, 간조직은 $0.6{\pm}0.2{\%}$로 비장에 비해 간 조직이 낮게 나타났다. Proteomoics 결과에서 방사선 내성 조직인 간은 ROS대사에 관여되는 단백질인 glutathione Stransferase Pi, carbonic anhydrase, NADH dehydrogenase, peroxiredoxin VI, riken cDNA 등이 방사선 조사 후 증가되었고 apoptosis 관련된 단백질인 cytochrome c는 간과 비장 조직에서 확인되었다. 그러나 방사선 민감 조직인 비장에서는 방사선 조사 후 산화적 Stress에 관련된 단백질, apoptosis 관련 단백질, 신호 전달에 관련된 단백질, 면역반응, cell cycle, Ca 신호 전달, 대사 cycle에 관련된 단백질 등이 방사선에 관련하여 발현의 변화를 보여 주었다. 결론 : Apoptosis유도 수준이 다른 조직에서 apoptosis에 관련된 단백질과 redox에 관련된 단백질은 방사성 감수성 조절에 관련된 것으로 보인다.

• Pediatric Gastroenterology, Hepatology & Nutrition
• /
• 제27권1호
• /
• pp.15-25
• /
• 2024

Purpose: This study aimed to investigate the presence of autoantigens in the gastric juices of children. Methods: Gastric juice and serum samples were obtained from 53 children <15 years of age who underwent gastric endoscopy. Among these, 8, 22, and 23 participants were in the age groups 0-5, 6-10, and 11-15 years, respectively. These samples were analyzed using two-dimensional electrophoresis (2-DE), immunoblot analysis, and matrix-assisted laser desorption ionization-time of-flight mass spectrometry. Furthermore, we reviewed the histopathological findings and urease test results and compared them with the results of 2-DE and immunoblot analysis. Results: There were no statistically significant differences in urease test positivity, grades of chronic gastritis, active gastritis, or Helicobacter pylori infiltration of the antrum and body among the three age groups. Three distinct patterns of gastric juice were observed on 2-DE. Pattern I was the most common, and pattern III was not observed below the age of 5 years. Histopathological findings were significantly different among active gastritis (p=0.037) and H. pylori infiltration (p=0.060) in the gastric body. The immunoblots showed large spots at an approximate pH of 3-4 and molecular weights of 31-45 kDa. These distinct, large positive spots were identified as gastric lipase and pepsin A and C. Conclusion: Three enzymes, which are normally secreted under acidic conditions were identified as autoantigens. Further investigation of the pathophysiology and function of autoantigens in the stomach is required.

• 한국작물학회지
• /
• 제64권4호
• /
• pp.422-431
• /
• 2019

본 연구는 옥수수 유수형성기(본 잎 6개)에 10일 동안 수분부족처리를 하였을 때 F₁ 옥수수 교잡종 식물체의 생리적 반응과 프로테옴 변화를 분석한 것이다. 실험에 사용한 품종은 일미찰과 광평옥이었다. 1. 정상구에 비해 수분이 결핍된 옥수수 교잡종에서는 평균 3개의 잎이 감소했고, 잎 면적은 각각 32~34 % 감소했으며, 경장은 일미찰에서는 약 14%, 광평옥에서는 약 27% 줄었다. 웅수 길이는 일미찰과 광평옥에서 각각 74, 82%가 감소하였다. 2. V4~6 엽기 때 10일간의 수분 결핍 처리는 옥수수의 모든 부분에서 건물중을 감소시켰으며 특히, 줄기의 건물중이 잎과 뿌리보다 훨씬 감소하였다. 일미찰과 광평옥에서 잎과 줄기의 건물중은 각각 약 83%, 73% 감소했으며, 수술 건물중은 각각 약 35, 86% 감소했다. 3. V4~6 엽기 때 10일간의 수분 결핍 처리는 옥수수의 모든 부분에서 건물중을 감소시켰으며 특히, 줄기의 건물중이 잎과 뿌리보다 훨씬 감소하였다. 일미찰과 광평옥에서 잎과 줄기의 건물중은 각각 약 83%, 73% 감소했으며, 수술 건물중은 각각 약 35, 86% 감소했다. 4. 이차원전기영동방법으로 정상구와 한발 스트레스를 받은 교잡종에서 다른 단백질 발현양상을 나타내는 21 개의 단백질 spot을 확인하였다. MALDI-TOF MS (matrix assisted laser desorption ionization-time of flight mass spectrometry) 및 단백질 데이터베이스 분석을 통해 21개의 단백질 spot 중 탄수화물 대사에 관련된 단백질이 8개, 스트레스 관련 단백질이 6개, 지방산 이화작용 및 광합성에 관련된 단백질이 각각 2개, 에너지 대사 및 수송에 관련된 단백질이 각각 1개가 분석되었다. 5. 이들 단백질 중 Triosephosphate isomerase, fructose-bisphosphate aldolase, uncharacterized protein을 제외하고 한발 스트레스 처리시 일미찰과 광평옥 모두에서 단백질 발현양이 증가하였으며, lactoylglutathione lyase, delta 3,5-delta 2,4-dienoyl-CoA isomerase은 광평옥에서만 과발현되었다.

• 한국식품위생안전성학회지
• /
• 제32권4호
• /
• pp.284-289
• /
• 2017

본 연구는 국내 미지정 피막제인 산화형 폴리에틸렌 왁스를 분석하기 위해 다양한 분석방법을 비교, 검토하였으며, 확립된 최적 분석법을 이용하여 시중 유통되는 과일 및 견과류에서 산화형 폴리에틸렌 왁스의 사용여부를 모니터링 하였다. 최적 분석법을 확립하기 위하여 산가, HT-GPC, MALDI-TOF/MS, GC-FID, FT-IR 등을 비교하였다. 산가측정 결과 산화형 폴리에틸렌 왁스를 첨가한 시료와 첨가하지 않은 시료 모두 산가가 1.12 mg KOH/g로 산가의 변화가 나타나지 않아 산가를 이용한 식품 중 산화형 폴리에틸렌 왁스의 분석은 어려울 것으로 판단되었으며, HT-GPC분석은 표준물질의 분산도 값이 약 3.4로 확인되었으며, GPC를 이용한 분석은 실험실간 낮은 재현성과 컬럼 등 분석조건에 따른 결과 값이 달라질 수 있기 때문에 분자량의 분산도만으로 식품 중 산화형 폴리에틸렌 왁스의 분석은 어려울 것으로 판단되었다. MALDI-TOF/MS 분석은 700~2,000 m/z에 peak가 관찰되었지만, 산화형 폴리에틸렌 왁스 성적서에 적혀있는 분자량 범위인 2,700~2,800 m/z와 큰 차이를 보여, MALDI-TOF/MS를 이용한 분석 역시 어려울 것으로 판단되었다. GC-FID를 이용한 지방산 분석결과 산화형 폴리에틸렌 왁스 표준물질에서 과일 및 견과류에서 흔히 볼 수 없는 지방산인 C11:0, C15:1 지방산이 검출되었지만, 제외국 허용기준으로 첨가하여 분석하였을 때 관찰되지 않고, 고농도를 첨가할 경우에만 피크가 관찰되어 GC-FID를 이용한 분석은 어려울 것으로 판단되었다. FT-IR 분석결과 코팅된 견과류 시료와 코팅되지 않은 견과류 시료를 비교하였을 때 코팅된 시료에서 산화형 폴리에틸렌 왁스 표준물질의 spectrum과 일치하는 spectrum을 나타내어, 식품 중 산화형 폴리에틸렌 왁스의 분석은 FT-IR로 분석이 가능함을 확인하였다. 확립된 FT-IR 분석법을 이용하여 국내외 대형마트에서 구입한 111건의 과일 및 견과류를 분석한 결과 산화형 폴리에틸렌 왁스를 사용한 시료가 없음을 확인하였다.