• 제목/요약/키워드: expression profile analysis

검색결과 301건 처리시간 0.028초

Detection of Matrix Metalloprotease-9 and Analysis of Protein Patterns in Bovine Vaginal Mucus during Estrus and Pregnancy

  • Kim, Sang-Hwan;Baek, Jun-Seok;Lee, Ho-Jun;Min, Kwan-Sik;Lee, Deuk-Hwan;Yoon, Jong-Taek
    • 한국수정란이식학회지
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    • 제27권2호
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    • pp.93-100
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    • 2012
  • To investigate the biochemical nature of changes in vaginal physiology during estrus and pregnancy, we examined the cytology and viscosity, and monitored the protein expression profile in vaginal mucus during estrus and pregnancy. The viscosity progressively decreased from estrus to pregnancy. Cell type analysis revealed that white blood cells progressively increased from estrus to pregnancy, while red blood cells progressively decreased during pregnancy. The cornification index (CI) was higher in estrus than in pregnancy. Protein mass spectrumetry identified the presence of ribosome-binding protein 1, GRIP 1 (Glutamate receptor-interacting protein 1)-associated protein 1, DUF729 (Domain of unknown function729) domain-containing protein 1, prolactin precursor, dihydrofolatereductase, and MMP (Matrix metalloprotease)-9 in vaginal mucus. MMP-2 and MMP-9 proteins in the vaginal mucus were active throughout estrus and gestation, as measured by a gelatinase assay, but most abundant in the vaginal mucus on day 0 of estrus. Results from ELISA of MMP-2 and MMP-9 were in accordance with the gelatinase assay. In light of the crucial role of metalloproteinases in extracellular matrix remodeling, the level of MMP-9 in vaginal mucus might be useful as an indicator of estrus and pregnancy to increase the efficiency of reproduction.

Molecular Characterization, Chromosomal Localizations, Expression Profile, and Association Analysis of the Porcine PECI Gene with Carcass Traits

  • Gao, H.;Fan, B.;Zhu, M.J.;Liu, Bang
    • Asian-Australasian Journal of Animal Sciences
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    • 제23권1호
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    • pp.7-12
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    • 2010
  • The full-length cDNA of the porcine peroxisomal ${\Delta}^3$,${\Delta}^2$-enoyl-CoA isomerase (PECI) gene encodes a monofunctional peroxisomal ${\Delta}^3$,${\Delta}^2$-enoyl-CoA isomerase. Cloning and sequencing of the porcine PECI cDNA revealed the presence of an 1185-base pair open reading frame predicted to encode a 394-amino acid protein by the 5'rapid amplification of cDNA ends (5'RACE) and EST sequences. The porcine PECI gene was expressed in seven tissues (heart, liver, spleen, lung, kidney, skeletal muscle, fat) which was revealed by reverse transcriptase-polymerase chain reaction (RT-PCR). The porcine PECI was mapped to SSC71/2 p11-13 using the somatic cell hybrid panel (SCHP) and the radiation hybrid panel (RH) (LOD score 12.84). The data showed that PECI was closely linked to marker S0383. A C/T single nucleotide polymorphism in PECI exon 10 (3'UTR) was detected as a PvuII PCR-RFLP. Association analysis in our experimental pig population showed that different genotypes of PECI gene were significantly associated with the Average Backfat thickness (ABF) (p<0.05) and Buttock backfat thickness (p<0.01).

Selective miRNA Expression Profile in Chronic Myeloid Leukemia K562 Cell-derived Exosomes

  • Feng, Dan-Qin;Huang, Bo;Li, Jing;Liu, Jing;Chen, Xi-Min;Xu, Yan-Mei;Chen, Xin;Zhang, Hai-Bin;Hu, Long-Hua;Wang, Xiao-Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권12호
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    • pp.7501-7508
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    • 2013
  • Background: Chronic myeloid leukemia (CML) is a myeloproliferative disorder of hematopoietic stem cell scarrying the Philadelphia (Ph) chromosome and an oncogenic BCR-ABL1 fusion gene. The tyrosine kinase inhibitor (TKI) of BCR-ABL1 kinase is a treatment of choice for control of CML. Objective: Recent studies have demonstrated that miRNAs within exosomes from cancer cells play crucial roles in initiation and progression. This study was performed to assess miRNAs within exosomes of K562 cells. Methods: miRNA microarray analysis of K562 cells and K562 cell-derived exosomes was conducted with the 6th generation miRCURYTM LNA Array (v.16.0). Gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were also carried out. GO terms and signaling pathways were categorized into 66 classes (including homophilic cell adhesion, negative regulation of apoptotic process, cell adhesion) and 26 signaling pathways (such as Wnt). Results: In exosomes, 49 miRNAs were up regulated as compared to K562 cells, and two of them were further confirmed by quantitative real-time PCR. There are differentially expressed miRNAs between K562 cell derived-exosomes and K562 cells. Conclusion: Selectively expressed miRNAs in exosomes may promote the development of CML via effects on interactions (e.g. adhesion) of CML cells with their microenvironment.

Protein Profile of Embryonic Stem Cells Following Differentiation

  • D. I. Jin;Kang, H. S;Kim, H. J.;Lee, S. H.;Park, C. S.;K. S. Im;Lee, H. M.
    • 한국가축번식학회지
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    • 제26권4호
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    • pp.377-384
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    • 2002
  • To investigate the expression patterns of proteins and growth factor signals in differentiated rabbit embryonic stem (ES) cells, ES cells with confluent stage grown of feeder layer and differentiated cells into embryoid bodies (EB) without feeder cell were applied to protein gel and Western blotting analysis. There were 66kDa and 28kDa specifically expressed in differentiated ES cell but not in undifferentiated ES cell while 25kDa protein band showed up in only undifferentiated ES cells. Also there were some difference of protein bands in several area of gel between differentiated and undifferentiated ES cells such as about 100 kDa, 50kDa and 27kDa areas, but there was no difference in band pattern of one-dimensional gel analysis between mouse ES cells and rabbit ES cells. IGF-I receptor and EGF receptor were expressed in differentiated cells and undifferentiated cells. And ICF-I and EGF were not expressed in both differentiated and undifferentiated cells. These results indicated that ES cells express their own proteins to inhibit differentiation while EB cells synthesize different proteins to differentiate, and 16F-I receptor and EGF receptor were expressed in both ES and EB cells probably for the different functions.

A Comprehensive Identification of Synaptic Vesicle Proteins in Rat Brains by cRPLC/MS-MS and 2DE/MALDI-TOF-MS

  • Lee, Won-Kyu;Kim, Hye-Jung;Min, Hye-Ki;Kang, Un-Beom;Lee, Cheol-Ju;Lee, Sang-Won;Kim, Ick-Young;Lee, Seung-Taek;Kwon, Oh-Seung;Yu, Yeon-Gyu
    • Bulletin of the Korean Chemical Society
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    • 제28권9호
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    • pp.1499-1509
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    • 2007
  • Proteomic analyses of synaptic vesicle fraction from rat brain have been performed for the better understanding of vesicle regulation and signal transmission. Two different approaches were applied to identify proteins in synaptic vesicle fraction. First, the isolated synaptic vesicle proteins were treated with trypsin, and the resulting peptides were analyzed using a high-pressure capillary reversed phase liquid chromatography/tandem mass spectrometry (cRPLC/MS/MS). Alternatively, proteins were separated by two-dimensional gel electrophoresis (2DE) and identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-TOF/MS). Total 18 and 52 proteins were identified from cRPLC/MS-MS and 2DE-MALDI-TOF-MS analysis, respectively. Among them only 2 proteins were identified by both methods. Of the proteins identified, 70% were soluble proteins and 30% were membrane proteins. They were categorized by their functions in vesicle trafficking and biogenesis, energy metabolism, signal transduction, transport and unknown functions. Among them, 27 proteins were not previously reported as synaptic proteins. The cellular functions of unknown proteins were estimated from the analysis of domain structure, expression profile and predicted interaction partners.

호로파(葫蘆巴) 종자 추출물이 난소 절제술을 받은 쥐에서 갱년기 증상 및 골다공증에 미치는 영향 (Trigonella Goenum-Graecum Seed Extract Alleviates Menopausal Symptoms and Osteoporosis in Ovariectomized Rats)

  • 김민주;신미래;최학주;노성수
    • 대한본초학회지
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    • 제39권3호
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    • pp.85-95
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    • 2024
  • Objectives : We proposed the hypothesis that administering Trigonella foenum-graecum seed extract (TSE) could alleviate menopausal symptoms and osteoporosis resulting from estrogen deficiency. Methods : Ovariectomized (OVX) rats were administered TSE at doses of 300 or 600 mg/kg body weight for 8 weeks, followed by measurement of serum lipid profile and serum bone markers using ELISA kits. Additionally, analysis of related genes in the femur and uterus was performed using Western blot and real-time PCR. Additionally, micro-CT analysis was performed to investigate the protective effect of TSE against bone loss due to oophorectomy. Results : The administration of TSE led to significant reductions in triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL) cholesterol, and glucose levels in the serum of OVX rats. Furthermore, TSE increased estradiol levels in the serum and notably improved the levels of biochemical markers associated with bone metabolism. Additionally, TSE exerted significant regulatory effects on the mRNA levels of alkaline phosphatase (ALP) and receptor activator of nuclear factor kappa-B ligand (RANKL)-genes closely associated with bone metabolism in the femur. TSE also demonstrated pronounced effects on uterine tissue, with improvements observed in gene expression related to estrogen receptors. Conclusion : Our findings confirm the efficacy of TSE in ameliorating menopause symptoms by modulating elements associated with both bone and lipid metabolism in the serum, uterine tissue, and femur of OVX rats. The present findings suggest that TSE may offer potential therapeutic effects for symptoms related to menopause and osteoporosis in females.

제조방법에 따른 떫은감 (Diosyros kaki Thumb.)의 대사체 프로파일링과 중성지질/콜레스테롤 대사 관련 유전자발현 연구 : in vitro 및 in vivo 연구 (Metabolites profiling and hypolipidemic/hypocholesterolemic effects of persimmon (Diosyros kaki Thumb.) by different processing procedures: in vitro and in vivo studies)

  • 박수연;오은경;임예니;신지윤;정희아;박송이;이진희;최정숙;권오란
    • Journal of Nutrition and Health
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    • 제51권4호
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    • pp.275-286
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    • 2018
  • 본 연구는 떫은감 청도반시의 분말을 열수 추출하고 칼럼 추출하는 과정에서 획득한 PWE와 TEP에 대한 대사체 프로파일을 분석하고, 유리지방산을 처리한 HepG2 세포와 식이로 고지혈증을 유도한 Wistar계 흰쥐의 간조직을 사용하여 중성지질 및 콜레스테롤 대사 관련 유전자 발현을 측정하였다. 대사체 분석은 가스크로마토그래프-질량분석법과 액체크로마토그래프-질량분석법을 사용하였으며, PLS-DA 분석과 heatmap 분석을 실시한 결과 PWE는 떫은감 분말과 비교적 유사한 대사체 프로파일을, 그리고 TEP는 떫은감 분말과 매우 다른 대사체 프로파일을 가지고 있음을 확인하였다. 세포실험과 또한 세포실험에서 얻어진 결과를 검증하기 위해 수행된 동물실험에서 PWE와 TEP는 모두 SREBP1c와 FAS 유전자 발현을 감소하여 간의 지방축적을 감소시키는 것으로 관찰되었으나, 콜레스테롤 축적을 억제하는 효능은 PWE에 비해 TEP에서 우세하게 증가하는 것이 관찰되었다. 특별히 TEP는 SREBP2, HMGCR 유전자 발현을 억제하고 LDLR 유전자 발현을 촉진하는 것으로 나타났다. TEP는 탄닌 중 Gallic acid 그리고 장쇄지방산아미드인 Oleamide와 Palmitamide 함량이 유의하게 증가되었으므로, 향후 이들 성분과 타깃 유전자의 상관성을 분석하고, 이를 통해 시스템네트워크로 나타내어 대사체 프로파일에 따른 지질 및 콜레스테롤 대사 간 상호 영향을 추가적으로 연구해야 할 것으로 사료된다.

Isolation and characterization of LHT-type plant amino acid transporter gene from Panax ginseng Meyer

  • Zhang, Ru;Zhu, Jie;Cao, Hong-Zhe;Xie, Xiao-Lei;Huang, Jing-Jia;Chen, Xiang-Hui;Luo, Zhi-Yong
    • Journal of Ginseng Research
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    • 제37권3호
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    • pp.361-370
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    • 2013
  • A lysine histidine transporter (LHT) cDNA was isolated and characterized from the roots of Panax ginseng, designated PgLHT. The cDNA is 1,865 bp with an open reading frame that codes for a protein with 449 amino acids and a calculated molecular mass of 50.6 kDa with a predicted isoelectric point of 8.87. Hydropathy analysis shows that PgLHT is an integral membrane protein with 9 putative membrane-spanning domains. Multiple sequence alignments show that PgLHT shares a high homology with other plant LHTs. The expression profile of the gene was investigated by real-time quantitative polymerase chain reaction during various chemical treatments. PgLHT was up-regulated in the presence of abscisic acid, salicylic acid, methyl jasmonate, NaCl, and amino acids. To further explore the function of PgLHT gene, full-length cDNA of PgLHT was introduced into P. ginseng by Agrobacterium rhizogenes A4. The overexpression of PgLHT in the hairy roots led to an obviously increase of biomass compared to the controls, and after addition of the amino acids, the overexpressed-PgLHT hairy roots grew more rapidly than untreated controls during early stage of the culture cycle. The results suggested that the PgLHT isolated from ginseng might have role in the environmental stresses and growth response.

Acquired JAK-2 V617F Mutational Analysis in Pakistani Patients with Essential Thrombocythemia

  • Sultan, Sadia;Irfan, Syed Mohammed
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권16호
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    • pp.7327-7330
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    • 2015
  • Background: Essential thrombocythemia (ET) is a clonal hemopoietic stem cell myeloproliferative neoplasm characterized by persistent thrombocytosis along with megakaryocytic hyperplasia. In the last decade following the identification of an acquired JAK2 V617F mutation, there has been acceleration in our understanding of this disease. The rational of this study was to determine the mutational profile of JAK2 V617F in Pakistan patients with ET. Materials and Methods: In this retrospective cross sectional study, 21 patients with ET were enrolled from January 2011 to December 2014. Patients were diagnosed based on WHO criteria for essential thrombocythemia. Complete blood count was done on an automated hematology analyzer, while JAK2 V617F expression was evaluated by polymerase chain reaction. Results: The mean age was $56.7{\pm}19.0$ years (range 18-87) and the male to female ratio was 1:1.1. The frequency of JAK2 V617F positivity in our ET patients was found to be 61.9%. The mean hemoglobin was $11.7{\pm}2.4$ g/dl with a total leukocyte count of $13.3{\pm}8.1{\times}109/l$ and a platelet count of $1188{\pm}522{\times}109/l$. Positive correlations for JAK2 V617F mutation were established with high TLC count and raised LDH (P<0.05). No correlation of JAK2 V617F could be established with age and gender (P>0.05). Conclusions: JAK2 V617F mutation frequency in our ET patients was similar to those reported previously. Screening for the mutation in all suspected essential thrombocythemia cases could be beneficial in differentiating patients with reactive and clonal thrombocytosis.

Proteomic Characterization of the 'Agakong', a Small-seeded Recombinant Inbred Line Derived from 'Eunhakong' (Glycine max) $\times$ 'KLG10084' (Glycine soja)

  • Choi, Ung-Kyu;Ryu, Hyun-Su;Kim, Hyun-Tae;Yun, Sun-Mi;Lee, Su-Jin;Choi, Jae-Dek;Hwang, Young-Hyun;Choi, Soo-Young;Kwon, Oh-Shin
    • Food Science and Biotechnology
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    • 제17권5호
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    • pp.912-918
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    • 2008
  • This study was conducted to identify the differences in proteomic characteristics of 'Agakong', recombinant inbred line, and its parental genotypes 'Eunhakong' (Glycine max) and 'KLG10084' (G. soja). The isoflavone content of 'Agakong' was 3 times higher than that of its parental lines. A combined high-throughput proteomic approach was employed to determine the expression profile and identity of proteins using 2-dimensional gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry. The overall distribution patterns of proteins are quite similar, but lots of protein spot intensities varied among the genotypes. A total of 41 proteins, representing significant difference in the quantities of protein among the lines, were successfully identified. Among them, more than 50% of the proteins identified were subunits of glycinin and $\beta$-conglycinin, 2 major storage proteins. This study showed that the proteomic analysis could help to define specific changes in protein level and composition, which can occur in the generation of new soybean varieties.