• 제목/요약/키워드: PBLs

검색결과 25건 처리시간 0.028초

단세포 전기영동법을 이용한 인체, 마우스 및 랫드 림프구의 방사선에 의해 유발된 DNA 손상 측정 (Analysis of gamma-ray-induced DNA damage in human, mouse and rat peripheral blood lymphocytes using single-cell gel electrophoresis)

  • 오헌;정우희;박혜란;김성호;조성기
    • 대한수의학회지
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    • 제44권1호
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    • pp.41-47
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    • 2004
  • The alkaline single-cell gel electrophoresis (SCGE) assay, called the comet assay, has been applied to detect DNA damage induced by a number of chemicals and biological factors in vivo and in vitro. The DNA damage was analysed by tail moment (TM) and tail length (TL), which were markers of DNA strand breaks in SCGE. Human, mouse and rat peripheral blood lymphocytes (PBLs) were irradiated with different doses of $^{60}Co$ ${\gamma}$-rays, e.g. 1, 2, 4, and 8 Gy at a dose rate of 1 Gy/min. A dose-dependent increase in TM (p<0.01) and TL (p<0.01) was obtained at all the radiation doses (1-8 Gy) in human, mouse and rat PBLs. Mouse PBLs were more sensitive than human PBLs which were in turn more sensitive than rat PBLs when the treated dosages were 1 and 2 Gy. However, human PBLs were more sensitive than mouse PBLs which were in turn more sensitive than rat PBLs when the irradiation dosages were 4 and 8 Gy. Data from all three species could be fitted to a linear-quadratic model. These results indicated that there may be inherent differences in the radio-sensitivity among PBLs of mammalian species.

마우스와 사람 림프구에서 방사선에 의한 미소핵의 형성 및 고려인삼의 효과 (Induction of Micronuclei in Human and Mouse Lymphocytes Irradiated with Gamma Radiation and Effect of Panax ginseng C.A. Meyer)

  • 김성호;오헌;이송은;이윤실;김태환;정규식;류시윤
    • Journal of Radiation Protection and Research
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    • 제22권3호
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    • pp.153-160
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    • 1997
  • 사람의 말초혈액림프구와 C57BL/6마우스의 비장림프구를 사용하여 시험관내에서 감마선을 조사하고 배양하여 세포질분열 차단 림프구내에 형성되는 미소핵의 빈도를 측정하였다. 미소핵 발생빈도는 방사선조사 선량에 비례하여 증가하였으며 linen-quadratic 곡선식에 적용하여, 세포 당 0.2개의 미소핵이 유도되는 방사선량을 산출하면 사람의 말초혈액 림프구에 비하여 마우스 비장림프구에서 1.67배 민감하였다. 미소핵시험방법을 이용하여, 사람의 말초혈액 림프구에 대한 인삼의 방사선 방호효과를 시험관내 시험으로, 마우스의 미장림프구에 대한 효과를 생체내 시험으로 검정하였다. 사람림프구에 있어서 방사선(3Gy)에 의해 유도되는 미소핵의 수는 방사선조사 전 및 후 투여군에서 공히 감소하였으며(p<0.01), 마우스를 사용한 생체시험에서도 림프구의 미소핵 발생빈도는 낮게 관찰되었다(p<0.025). 이상의 결과에서 인삼은 인체에서도 방사선에 의한 세포장해를 감소시킬 가능성을 나타냈다.

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Transcriptional Profiles of Peripheral Blood Leukocytes Identify Patients with Cholangiocarcinoma and Predict Outcome

  • Subimerb, Chutima;Wongkham, Chaisiri;Khuntikeo, Narong;Leelayuwat, Chanvit;McGrath, Michael S.;Wongkham, Sopit
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권10호
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    • pp.4217-4224
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    • 2014
  • Cholangiocarcinoma (CCA), a slow growing but highly metastatic tumor, is highly prevalent in Northeast Thailand. Specific tests that predict prognosis of CCA remain elusive. The present study was designed to investigate whether peripheral blood leukocyte (PBL) transcriptional profiles might be of use as a prognostic test in CCA patients. Gene expression profiles of PBLs from 9 CCA and 8 healthy subjects were conducted using the Affymetrix HG_U133 Plus 2.0 GeneChip. We indentified informative PBLs gene expression profiles that could reliably distinguish CCA patients from healthy subjects. Of these CCA specific genes, 117 genes were up regulated and 60 were down regulated. The molecular and cellular functions predicted for these CCA specific genes according to the Gene Ontology database indicated differential PBL expression of host immune response and tumor progression genes (EREG, TGF ${\beta}1$, CXCL2, CXCL3, IL-8, and VEGFA). The expression levels of 9 differentially expressed genes were verified in 36 CCA vs 20 healthy subjects. A set of three tumor invasion related genes (PLAU, CTSL and SERPINB2) computed as "prognostic index" was found to be an independent and statistically significant predictor for CCA patient survival. The present study shows that CCA PBLs may serve as disease predictive clinically accessible surrogates for indentifying expressed genes reflective of CCA disease severity.

DNA Repair Capacity in Peripheral Blood Lymphocytes Predicts Efficacy of Platinum-based Chemotherapy in Patients with Gastric Cancer

  • Zhang, Yi-Yin;Gu, Kang-Sheng
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권9호
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    • pp.5507-5512
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    • 2013
  • Objective: To investigate the correlation between ERCC1 expression levels in tumor tissue and peripheral blood lymphocytes (PBL) from patients with gastric cancer and assess the relationship between PBL DNA repair rate (DRR) and the efficacy of platinum chemotherapy. Methods: A total of 53 patients with gastric cancer receiving surgery and 20 controls were studied. ERCC1 protein expression in tumour tissue and PBL were determined by immunohistochemical staining. The PBL DRRs of 47 advanced patients and 20 controls were estimated by comet assay. Results: The positive expression rates of ERCC1 were 67. 9%, 56. 6% and 10.0% in tumour tissues, PBLs of gastric cancer patients, and PBLs of the control group. PBL ERCC1 expression correlated with that in tissue (${\chi}^2$=15. 463, p=0.000). Pearson contingency coefficient=0.475). DRRs of cancer patients by tail length (TL) (Z=4. 662, p=0.000) and tail moment (TM) (Z=3. 827, p=0.000) were significantly lower than that of control group. When TL was applied as an indicator, the correlation between DRR and chemotherapy efficacy was significant (Spearman rank correlation r=0.327, p=0.032). Patients with low levels of DRR in PBL presented better short-term efficacy of chemotherapy than those with high levels of DRR. Conclusions: The ERCC1 expression in PBLs may indirectly reflect ERCC1 expression in gastric cancer tissues. Compared with non-cancer populations, patients with gastric cancer may have lower DNA repair capacity. DRR in PBL may predict the short-term efficacy of platinum-based chemotherapy for patients with advanced gastric cancer.

RNA aptamer 발현을 통한 CD4+ peripheral blood lymphocytes에서의 인간 면역결핍 바이러스의 증식 억제 (Inhibition of HIV-1 Replication in CD4+ Peripheral Blood Lymphocytes by Intracellular Expression of RNA Aptamer)

  • 이성욱
    • 미생물학회지
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    • 제39권4호
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    • pp.235-241
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    • 2003
  • 제1형 인간 면역결핍 바이러스 (human immunodeficiency virus type 1, HIV-1)의 Rev 단백질에 대하여 야생형보다 10배 더 잘 결합하도록 시험관에서 선택된 RRE40라 명명된 RNA aptamer가 과연 임상적으로 유용한지 알기 위하여 인체의 CD4^+ peripheral blood lymphocytes 세포에서 레트로바이러스 벡터를 이용하여 RRE40 RNA를 발현한 후에 그 세포에서의 HIV-1 증식 현상을 관찰하였다. 그 결과 대조군인 tRNA를 발현하는 유전자가 전달된 세포에 비해 RRE40 RNA를 발현하는 세포에서 보다 더 효과적으로 HIV-1의 증식이 억제되었다. 그러나 바이러스의 증식이 완전히 억제되지는 못 하였고 일시적 또는 감소된 형태로 바이러스 증식이 억제되었다. 이러한 결과는 RRE40 RNA가 decoy로서 세포에서의 HIV-1 증식 억제에 유용함을 시사하지만 RNA decoy를 HIV-1 감염 환자의 치료에 이용하기 위해선 보다 효과적인 유전자 전달방법 및 보다 개선된 RNA decoy의 개발 등이 필요할 것이다.

Cloning and Characterization of DAP10 homologue gene from Olive Flounder, Paralichthys olivaceus

  • 박찬일;김무찬;황지윤;김기혁;김주원
    • 한국어병학회지
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    • 제19권3호
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    • pp.227-233
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    • 2006
  • Olive flounder immunoreceptor DAP10 homologue cDNA was cloned from a peripheral blood lymphocytes (PBLs) cDNA library. The length of the olive flounder DAP10 cDNA is 473bp and it contains an open reading frame of 234bp. The predicted polypeptide sequence is 78 amino acids, consisting of a 22-amino acid leader, an 11-amino acid extracellular domain, a 21-amino acid transmembrane segment, and a 24-amino acid cytoplasmic domain. The amino acid sequence of olive flounder DAP10 has 56%, 50%, 32%, 31%, and 31% sequence identity with zebrafish DAP10, catfish DAP10, cattle DAP10, rat DAP10 and Monkey DAP10, respectively. Olive flounder DAP10 has a conserved aspartic acid in the transmembrane domain and a phophatidylinositol-3 kinase-binding site (YxxM/V) in the cytoplasmic region. Genomic organization reveals that olive flounder DAP10 comprises five exons and four introns. A phylogenetic analysis based on the deduced amino acid sequence grouped the olive flounder DAP10 with other species DAP10. In RT-PCR analysis, DAP10 transcripts were detected predominantly in PBLs, kidney, spleen and intestine.

Molecular identification and expression analysis of a natural killer enhancing factor-A from black rockfish Sebastes schlegelii

  • Lee, Jeong-Ho;Kim, Joo-Won;Park, Chan-Il
    • 한국어병학회지
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    • 제22권3호
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    • pp.343-352
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    • 2009
  • Natural-killer-cell-enhancing factor (NKEF) belongs to the newly defined peroxiredoxin (Prx) family. It was originally isolated from human erythroid cells. The black rockfish NKEF cDNA was identified through the expressed sequence tag (EST) analysis of PBLs libraries. The full-length NKEF cDNA was 1433 bp long and contained an open reading frame (ORF) of 594 bp that encoded 198 amino-acid residues. The 5' UTR had a length of 39 bp, and the 3’UTR 800 bp. The deduced amino-acid sequence of the black rockfish had a density 93.4, 92.9, 87.8, 85.8, 84.8, 83.8, 80.3, 79.7, 77.2, and 75.2% that of the pufferfish, olive flounder, channel catfish, zebrafish, chicken, common carp, Myotis lucifugus, cattle, human PrxI, rat PrxI, human NKEF-A, and Xenopus tropicalis, respectively. The NKEF gene was expressed in all the tissues of the black rockfish. The RT-PCR indicated that the NKEF transcripts were predominantly in the spleen and gill, less dominantly in the PBLs, head kidney, trunk kidney, and liver, and least in the intestine and muscles. This is the first report on the existence of the NKEF-A gene in black rockfish.

IMMUNOREGULATORY EFFECTS OF A MONOCLONAL ANTIBODY TO HUMAN 4-1 BB MOLECULE ON ALLOANTIGEN-MEDIATED IMMUNE RESPONSES.

  • Kim, Joong-Gon;Lee, Soo-Hyun;Lee, Jae-Woo;B.S. Kwon;Kang, Chang-Yuil
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1995년도 춘계학술대회
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    • pp.82-82
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    • 1995
  • 4-lBB molecule is expressed on the surface of activated CD4$\^$+/ and CD8$\^$+/ T cells. We generated a panel of anti-4-1 B5 murine mAbs using a fusion protein consisting of the extracellular domain of human 4-1 BB fused to Glutathione S-transferase. The binding activity against cell surface 4-1 BB molecule was assessed by flow cytometry analysis. These studies showed that several anti-4-1 BB mAbs bound to 10-30% of CD4$\^$+/ and CD8$\^$+/T cells in PHA or Con A stimulated PBLs, although these mAbs interacted with only, l-2% of CD4$\^$+/ and CD8$\^$+/ T cells in normal PBLs, indicating the specificity of mAbs to the 4-l BB molecule on activated CD4$\^$+/ and CD8$\^$+/ T cells. Next, we examined the effect of an anti-4-l BB mAb (4B4-1-1) on allogeneic mixed lymphocyte reactions (MLRs). The data indicated that the antibody significantly inhibited the proliferative response at higher concentrations. When tested with several T cell mitogens, the antibody had no stimulatory or inhibitory effects on the mitogen-mediated T cell proliferation. These data suggest that 4-1 BB molecule may play a role in the regulation of antigen-mediated immune response.

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PBL수업에서 교육과정 편성 과제에 대한 동기 설계가 학습자의 교과흥미와 과제난이도 인식에 미치는 영향 (Effects of Motivational Design on Curriculum Organization Tasks on Learners' Subjects Interest and Task Difficulty Recognition in PBL)

  • 이은철
    • 한국콘텐츠학회논문지
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    • 제20권1호
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    • pp.334-344
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    • 2020
  • 본 연구는 PBL(Problem Based Learning) 수업에서 학생들에 수행하는 과제에 켈러의 ARCS에 의해 동기설계를 반영하였을 때, 학습자의 교과흥미와 과제난이도 인식 수준에 미치는 영향을 탐색하기 위해서 수행되었다. 이를 위해서 일반교직을 전공하는 대학생 79명을 대상으로 수행되었다. 연구를 위해서 실험집단과 비교집단을 구성하였다. 교과 흥미와 과제난이도 인식 수준의 사전 수준을 측정하기 위해 1차 PBL 과제를 수행하였다. 다음으로 동기 설계가 반영된 과제의 영향을 검증하기 위해 2차 PBL 활동 수행하였다. 실험집단은 동기설계가 반영된 과제를 수행하였고, 비교집단은 동기설계가 반영이 되지 않은 과제를 수행하였다. 2차 PBL 활동이 종료된 후에 교과흥미와 과제난이도 인식 수준을 측정하였다. 수집된 자료는 사전 수준을 공변량으로 선정하여 ANCOVA를 사용하여 자료를 분석하였다. 그 결과 실험집단의 교과흥미 수준이 통계적으로 유의미하게 향상되었고, 과제난이도 인식 수준은 통계적으로 유의미하게 낮아진 것으로 검증되었다.

Molecular cloning and expression analysis of an interferon stimulated gene 15 from rock bream Oplegnathus fasciatus

  • Kim, Ju-Won;Kwon, Mun-Gyeong;Park, Myoung-Ae;Hwang, Jee-Youn;Park, Hyung-Jun;Baeck, Gun-Wook;Kim, Mu-Chan;Park, Chan-Il
    • 한국어병학회지
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    • 제23권2호
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    • pp.177-187
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    • 2010
  • The Interferon stimulated gene 15 (ISG15) is strongly induced in many cell types by IFNs, viral infections, and double-stranded RNA (poly I:C). The ISG15 homologue cDNA was isolated from the rock bream LPS stimulated leukocyte cDNA library. The rock bream ISG15 homologue was found to consist of 833 bp encoding 157 amino acid residues. Compared with other known ISG15 peptide sequences, the most conserved regions of the rock bream ISG15 peptide were found to be the tandem ubiquitin-like domains and a C-terminal LRLRGG conjugating motif, characteristic of mammalian and non-mammalian ISG15 proteins. Phylogenetic analysis based on the deduced amino acid sequence revealed a homologous relationship between the ISG15 sequence of rock bream and that of Atlantic salmon, Atlantic cod, northern snake head, black rockfish and olive flounder. The expression of the rock bream ISG15 molecule was induced in the peripheral blood leukocytes (PBLs) from 1 to 24 h following poly I:C stimulation, with a peak at 3 h post-stimulation. The rock bream ISG15 gene was predominantly expressed in the PBLs, spleen and gill.