• Title/Summary/Keyword: protein tissues

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Redistribution of Lactate Dehydrogenase Isozymes and Morphology of Tissues in Mus musculus after Irradiation (방사선 조사 후 생쥐(Mus musculus)조직의 형태와 젖산탈수소효소 동위효소의 재분포)

  • 박현도;염정주
    • Korean Journal of Environmental Biology
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    • v.17 no.3
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    • pp.263-270
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    • 1999
  • In order to identify the morphological changes of tissues in mouse after irradiation. We have observed the redistribution of LDH isozymes and the morphological changes of skeletal muscle, heart, kidney, liver and testis in mouse according to variation amount with the time after the 1 Gray and 3 Gray irradiation each. As a result of H-E (hematoxylin-eosin) stain, the apoptotic bodies were more easily observed in the liver than the other tissues and the quantity of the apoptotic bodies was proportionated to radiation amount. The number of apoptotic bodies was shown the highest at 1 day in most tissues and at 7 day in testis after irradiation. TUNEL (terminal deoxyribonucleodtidyl transferase-mediated dUTP-digoxigenin nick end labeling) staining was shown the same results as H-E staining. After the irradiation, the protein content was reduced in tissues except kidney. And protein content was reduced in all tissues at the initial period of 2 hours after 3 Gy irradiation. But it increased at 7 days after irradiation. LDH (EC 1.1.1.27, lactate dehydrogenase) activity was increased mostly in tissues at the early stage after 1 Gy irradiation. The maximum activity was detected earlier stage after 1 Gy irradiation than 3 Gy irradiation. The activity of LDH $A_4$ isozyme was decreased in the skeletal muscle, heart, kidney, and testis. The activity of $B_4$ and $A_2$$B_2$ sozyme was increased in the skeletal muscle and heart, and the activity of heterotetramer isozyme was increased in kidney The activity of $A_4$ isozyme in liver was detected high level and the activity of isozyme including subunit C elevated in testis. Therefore, LDH isozyme seems to play a role of lactate oxidase in most tissues except liver after irradiation. These data support that LDH isozyme is predomintly involved in the aerobic metabolism.

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Expression of the Proto-oncogene Pokemon in Colorectal Cancer - Inhibitory Effects of an siRNA

  • Zhao, Gan-Ting;Yang, Li-Juan;Li, Xi-Xia;Cui, Hui-Lin;Guo, Rui
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.4999-5005
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    • 2013
  • Objective: This study aimed to investigate expression of the proto-oncogene POK erythroid myeloid ontogenic factor (Pokemon) in colorectal cancer (CRC), and assess inhibitory effects of a small interference RNA (siRNA) expression vector in SW480 and SW620 cells. Methods: Semi-quantitative reverse transcription-polymerase chain reaction (PCR) and immunohistochemistry were performed to determine mRNA and protein expression levels of Pokemon in CRC tissues. Indirect immunofluorescence staining was applied to investigate the location of Pokemon in SW480 and SW620 cells. The siRNA expression vectors that were constructed to express a short hairpin RNA against Pokemon were transfected to the SW480 and SW620 cells with a liposome. Expression levels of Pokemon mRNA and protein were examined by real-time quantitative-fluorescent PCR and western blot analysis. The effects of Pokemon silencing on proliferation of SW480 and SW620 cells were evaluated with reference to growth curves with MTT assays. Results: The mRNA expression level of Pokemon in tumor tissues ($0.845{\pm}0.344$) was significantly higher than that in adjacent tumor specimens ($0.321{\pm}0.197$). The positive expression ratio of Pokemon protein in CRC (87.0%) was significantly higher than that in the adjacent tissues (19.6%). Strong fluorescence staining of Pokemon protein was observed in the cytoplasm of the SW480 and SW620 cells. The inhibition ratios of Pokemon mRNA and protein in the SW480 cells were 83.1% and 73.5% at 48 and 72 h, respectively, compared with those of the negative control cells with the siRNA. In the SW620 cells, the inhibition ratios of Pokemon mRNA and protein were 76.3% and 68.7% at 48 and 72 h, respectively. MTT showed that Pokemon gene silencing inhibited the proliferation of SW480 and SW620 cells. Conclusion: Overexpression of Pokemon in CRC may have a function in carcinogenesis and progression. siRNA expression vectors could effectively inhibit mRNA and protein expression of Pokemon in SW480 and SW620 cells, thereby reducing malignant cell proliferation.

Effects of Polygala tenuipolia on Expression of Fos-protein and Ethanol Amnesia in Rat

  • Lee, Soon-Chul;Kim, Kwang-Kyu;Jang, Jin-Hee;You, Kwan-Hee
    • Biomedical Science Letters
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    • v.8 no.3
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    • pp.167-172
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    • 2002
  • Effect of single administration of Polygala tenuipolia was examined on short-term memory in step through test and the intensity of the immunoreactive c-Fos protein induced by oral administration of ethanol. The acquisition of memory was significantly reduced by ethanol, and ethanol amnesia was remarkably reversed following oral administration of Polygala tenuifolia. c-Fos protein in normal rat brain was highly expressed in order of thalamus, pariental cortex, hippocampus, hypothalamus, amygdaloid and cingulate cortex. The expression of Fos protein was remarkably suppressed by single administration of ethanol. The inhibitory effect of ethanol on expression of Fos protein was reversed by single administration of Polygara tenuipolia, especially tissues of limbic areas such as amygdala, parietal cortex and CA3 of hippocampus. These results suggested that the amelioration process of Polygala tenuipolia on ethanol amnesia seems to be involve the expression of c-Fos protein in partly.

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TGF-β1 Protein Expression in Non-Small Cell Lung Cancers is Correlated with Prognosis

  • Huang, Ai-Li;Liu, Shu-Guang;Qi, Wen-Juan;Zhao, Yun-Fei;Li, Yu-Mei;Lei, Bin;Sheng, Wen-Jie;Shen, Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.19
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    • pp.8143-8147
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    • 2014
  • To investigate the expression intensity and prognostic significance of TGF-${\beta}1$ protein in non-small cell lung cancer (NSCLC), immunohistochemistry was carried out in 194 cases of NSCLC and 24 cases of normal lung tissues by SP methods. The PU (positive unit) value was used to assess the TGF-${\beta}1$ protein expression in systematically selected fields under the microscope with Leica Q500MC image analysis. We found that the TGF-${\beta}1$ PU value was nearly two-fold higher in NSCLC than in normal lung tissues (p=0.000), being associated with TNM stages (p=0.000) and lymph node metastases (p=0.000), but not to patient age, gender, smoking history, tumor differentiation, histological subtype and tumor location (P>0.05). Univariate analysis indicated that patients with high TGF-${\beta}1$ protein expression and lymph node metastases demonstrated a poor prognosis (both p=0.000,). Multivariate analysis showed that TGF-${\beta}1$ protein expression (RR = 2.565, p=0.002) and lymph node metastases (RR=1.874, p=0.030) were also independent prognostic factors. Thus, TGF-${\beta}1$ protein expression may be correlated to oncogenesis and serve as an independent prognostic biomarker for NSCLC.

Identification of Retinol-binding Protein Produced by Caprine Endometrium during Periattachment Period of Early Pregnancy

  • Liu, K.H.;Huang, J.C.;Lin, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.12
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    • pp.1708-1713
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    • 2002
  • Endometrial explants obtained from does between days 13 and 21 of pregnancy were cultured in a modified minimum essential medium in the presence of [$^35S$]methionine and [$^3H$]-leucine. Proteins synthesized and secreted into medium were analyzed by fluorography of two-dimensional polyacrylamide gel electrophoresis and fluorography. No marked qualitative changes in patterns of protein production by caprine endometrium between days 13-21 of pregnancy. At least 11 proteins showed consistently a clear spot or a grouping of spots with characteristic location on two-dimensional gels. A major low molecular weight protein consisted of two major isoforms (pI 5.3-6.0) of similar molecular mass (21 kDa). Limited N-terminal sequence analysis of these two isoforms showed that the protein had complete homology with bovine placental and plasma retinol-binding protein (RBP) over the first 20 amino acids. Through use of the antiserum raised against bovine placental RBP, immunoreactive RBP was detected in cultures conditioned by uterine explants prepared at days 13, 15 and 21 of pregnancy. In the present study, proteins synthesized and secreted by caprine endometrium during periattachment period of early pregnancy were characterized. The pregnant endometrium secreted a number of neutral-to-acidic proteins which constituted, in part, the histotroph. A vitamin A-transport protein, RBP, was identified in cultures conditioned by endometrium of days 13-21 of pregnancy. The uterine endometrium is the only source of retinol for embryonic tissues. The uterine RBP appears to transport retinol locally toward embryonic tissues. Secretion of RBP by caprine endometrium of days 13, 15 and 21 of pregnancy suggested that retinol played an important role in conceptus development during periattachment period of early pregnancy.

Induction of Leptin cDNA Expression in Esherichia coli Cells (대장균 세포에서 Leptin 유전자의 발현 유도)

  • 김은정;정인철;오상환;조무연
    • Journal of Life Science
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    • v.9 no.3
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    • pp.253-261
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    • 1999
  • Leptin gene, an obesity gene, has been known to involve in the regulation of food intake and body weight. It is also thought to be related to the glucose metabolism, insulin secretion and type II diabetes mellitus. Recently, the production of recombinant leptin protein has been attempted for the application in the treatment of obesity and the correction of hereditary obesity and type II diabetes. In the present study, leptin cDNA was cloned from mouse fat cells by RT-PCR and prokaryotic expression of leptin was attempted in order ot prepare a leptin-specific antigen. Immunization of a rabbit with the leptin-specific antigen into a rabbit resulted in the generation of leptin-specific antiserum that could be useful in the detection of leption expressed in various tissues. The sequence of leptin cDNA prepared in the present study wa identical to the previously reported one. Transformation of E. coli(DH5a) cells with the leptin cDNA-inserted translation vector, pGEX-4T-3-leptin followed by treatment with IPTG (0.1mM) resulted in the expression of a large amount of GST-leptin fusion protein with a molecular weight of 44 KDa as an inclusion body. Denaturation of the insoluble fusion protein by 8M urea, 6M guanidium-HCI or 0.1% 2-mercaptoethanol followed by a slow oxidation could not solubilize the inclusion body. The cell extract was subjected to SDS-PAGE and GST-leptin protein electroeluted from the gel was then injected into a rabbit subcutaneously for the immunization. Anti-GST-leptin rabbit antiserum which had a cross reactivity to the GST-leptin protein was generated. Leptin protein expressed in mouse brain and fat tissues was detected by Western blot immunodetection system using the antiserum generated in the present study.

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Cloning and Expression of Bovine Polyadenylate Binding Protein 1 cDNA in Mammary Tissues

  • Kim, J.H.;Jeon, D.H.;Choi, Y.J.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.6
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    • pp.771-776
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    • 2001
  • A pregnant-induced clone was identified by differential screening from a cDNA library of bovine mammary gland. The clone was identified as a cDNA encoding a polyadenylate binding protein 1 (PABP). The cDNA clone had a total length of 1,911 nucleotides coding for 636 amino acids. The nucleotide sequence of the bovine PABP was 95% and 94% identical to those of human and mouse species, respectively. Comparison of the deduced amino acid sequences of bovine PABP with those of human species showed 100% identity. Induction of the PABP mRNA was observed in bovine mammary tissues at pregnant 7 and 8 months compared to virgin, lactating and involuted states. Expression of the PABP gene was examined in mammary epithelial HC11 cells at proliferating, differentiated and apoptotic conditions. The mRNA levels of PABP gene were similar between proliferating and differentiated cells, but expression levels were very low in apoptotic cells compared to other conditions. Results demonstrate that the PABP gene is induced during pregnancy at which stage mammary epithelial cells are actively proliferating.

In situ Hybridization of a Megalocytivirus Using Nucleic Acid Probes against ATPase and the Major Capsid Protein of Rock Bream Iridovirus

  • Lee, Nam-Sil;Do, Jeong-Wan;Jung, Sung-Ju;Park, Mi-Seon;Kim, Jin-Woo;Kim, Yi-Cheong
    • Fisheries and Aquatic Sciences
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    • v.9 no.4
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    • pp.146-152
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    • 2006
  • Systemic infections of maricultured fishes by Megalocytivirus species have occurred over a broad area in South Korea, causing extensive economic loss. We developed digoxigenin-labeled nucleic acid probes against the 230-bp ATPase and 311-bp major capsid protein (MCP) of rock bream Oplegnathus fasciatus iridovirus (RBIV) using polymerase chain reaction, and an in situ hybridization (ISH) method to detect Megalocytivirus in formalin-fixed tissues of mariculture species (rock bream, sea bass, and olive flounder). ISH-positive cells were abundant in the hematopoietic and connective tissues of various organs, while brain tissue showed little or no signal. The ISH procedure can become an important diagnostic tool in complement with histopathological methods, and advances epidemiological studies on the origin and distribution of Megalocytivirus in mariculture.

The Overexpression of Oncogenic Nemo-like Kinase in Gastric Cancer (위암에서 새로운 종양원인 유전자 Nemo-like Kinase의 발현 증가)

  • Kim, Min Gyu;Jung, Kwang Hwa;Nam, Suk Woo
    • YAKHAK HOEJI
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    • v.56 no.6
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    • pp.358-363
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    • 2012
  • Nemo-like kinase (NLK), an evolutionarily conserved serine/threonine protein kinase, plays an important role in wide variety of developmental events. NLK phosphorylates T-cell factor/lymphoid enhancer factor (TCF/LEF) transcriptional complex and suppresses wnt signaling pathway through inhibition of ${\beta}$-catenin/TCF complex interaction. However, the function of NLK in gastric carcinogenesis has not been investigated. In the present study, we have examined whether the NLK gene is involved in the development and/or progression of gastric cancers. NLK expression was analyzed by immunohistochemical staining in 153 advanced gastric cancer specimens. Immunhistochemical analysis showed increased expression of NLK in 91 (59.5%) out of 153 gastric cancer specimens. Statistically, there was no significant relationship between altered expression of NLK protein and clinicopathological parameters, including tumor differentiation, location, lymph node metastasis. We identified that mRNA and protein expression of NLK was significantly up-regulated in human gastric cancer tissues compare to corresponding normal gastric tissues. In addition, we found that human gastric cancer cell lines exhibited relatively high expression of NLK, as compared with normal gastric cells. The results of this study suggest that aberrant regulation of NLK may contribute to the development or progression of gastric cancers and serve as a potential biomarker for advanced gastric cancer patients.