• Molecular & Cellular Toxicology
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• 제5권3호
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• pp.243-249
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• 2009

Melamine, which is used to produce melamine resin for various industrial applications, has a high nitrogen content by mass. For this reason, it has been illegally added to foods to increase their apparent protein content. In the present investigation, melamine-induced oxidative damage of human lymphocyte DNA was evaluated by Comet assay. The in vitro oxidative DNA damage caused by melamine increased in a dose-dependent manner. This DNA damage was significantly inhibited by treatment with ascorbate. Moreover, the traditional Korean medicinal herb, named Acanthopanax, red ginseng and green tea markedly reduced the DNA damage. Various edible plant extracts also inhibited melamine-induced oxidative DNA damage in vitro. Melamine enhanced intracellular ROS generation, and this effect was suppressed by treatment with various antioxidants.

• 생약학회지
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• 제39권2호
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• pp.150-154
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• 2008

Butein is a one of polyphenolic compound widely available in numerous plants. It has broad biological activities including antioxidant and anti-inflammatory activities, which contributed to its protective effects against cancer. Evidences that butein influence proliferation of tumor cells make it important to determine how butein affects cell death of various cancers. In this study, we show that butein, a phenolic compound, induces apoptosis in human T lymphoma jurkat cells. We found that treatment of cells with butein increased apoptosis in a dose- and time- dependent manner as determined by staining cells with Annexin V and 7AAD. There was no significant apoptotic cell death when normal lymphocytes and monocytes from healthy donor were treated with butein. We also found caspase-3 activity was increased during butein-induced apoptosis. The buteininduced apoptotic cell death was blocked by the treatment of cells with caspase-3 inhibitor. These results indicate that butein has the potential to provide an effective strategy against cancer with the advantage of being widely avalible.

• IMMUNE NETWORK
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• 제20권4호
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• pp.34.1-34.8
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• 2020

Cryopreservation and thawing of PBMCs are inevitable processes in expanding the scale of experiments in human immunology. Here, we carried out a fundamental study to investigate the detailed effects of PBMC cryopreservation and thawing on transcriptomes. We sorted Tregs from fresh and cryopreserved/thawed PBMCs from an identical donor and performed single-cell RNA-sequencing (scRNA-seq). We found that the cryopreservation and thawing process minimally affects the key molecular features of Tregs, including FOXP3. However, the cryopreserved and thawed sample had a specific cluster with up-regulation of genes for heat shock proteins. Caution may be warranted in interpreting the character of any cluster of cells with heat shock-related properties when cryopreserved and thawed samples are used for scRNA-seq.

• Molecular & Cellular Toxicology
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• 제2권4호
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• pp.229-235
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• 2006

Volatile organic compounds (VOCs) are a major component of urban air pollution. It is documented that low exposure levels of VOCs induce alterations in immune reactivity resulting in a subsequent higher risk for the development of allergic reactivity and asthma. Despite these facts, there are few reports on the affected primary target and the underlying effective causal mechanisms. So in this study, to better understand the risk of BTX (benzene, toluene and o-xylene) which are the major VOCs and to identify novel biomarkers on immune response to these VOCs exposure in human T lymphocytes, we performed the toxicogenomic study by analyzing of gene expression profiles using 35 k human oligo-microarray. BTX generated specific gene expression patterns in Jurkat cell line. By clustering analysis, we identified some genes as potential markers on immuno-modulating effects of BTX. Four genes of these, HLA-DOA, ITGB2, HMGA2 and 5TAT4 were the most significantly affected by BTX exposure. Thus, this study suggests that these differentially expressed immune genes may play an important role in the pathogenesis on BTX exposure and have significant potential as novel biomarkers of exposure, susceptibility and response to BTC.

• Animal cells and systems
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• 제8권2호
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• pp.125-133
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• 2004

We have extended our previous work that cross-linking CD4 molecules using specific MAb induced antigen nonspecific, MHC unrestricted killing of virally infected target cells by CD$4^+$We have extended our previous work that cross-linking CD$4^+$ molecules using specific MAb induced antigen nonspecific, MHC unrestricted killing of virally infected target cells by CD$4^+$ T cells. The killing activity of antibody activated CD$4^+$T cells was completely blocked by herbimycin A, a protein tyrosine kinase (PTK) inhibitor, but not by bisindolylamaleimide, a protein kinase C (PKC) inhibitor. Herbimycin A treated human or bovine peripheral blood CD$4^+$T cells lacked PTK activity and failed to kill virally infected target cells even after cross-linking of CD4 molecules. The CD$4^+$cross-linking failed to induce effector cell proliferation or the transcription of TNF${\beta}$ Upregulation of TNF${\beta}$ was induced by incubating the antibody activated effector cells with BHV-1 infected D17 target cells for 10 h. Anti-TNF${\beta}$ antibody partially abolished (13-44%) the direct effector cell-mediated antiviral cytotoxicity. However, this antibody neutralized 70 to 100% of antiviral activity of effector and target cell culture supernatants against BHV-1 infected D17 cells. The inhibition level of the antiviral activity by the antibody was dependent on the effector and target cell ratio. These results support the hypothesis that increased p$56^ICK enzyme activity in effector cells transduces a signal critical for effector cell recognition of viral glycoproteins expressed on the target cells. Following target cell recognition, lytic cytokines known to participate in target cell killing were produced. A better understanding of the killing activity displayed by CD$4^+$T lymphocytes following surface receptor cross-linking will provide insight into the mechanisms of cytotoxic activity directed toward virally-infected cells.T cells. The killing activity of antibody activated CD$4^+$T cells was completely blocked by herbimycin A, a protein tyrosine kinase (PTK) inhibitor, but not by bisindolylamaleimide, a protein kinase C (PKC) inhibitor. Herbimycin A treated human or bovine peripheral blood CD4T cells lacked PTK activity and failed to kill virally infected target cells even after cross-linking of CD4molecules. The CD4 cross-linking failed to induce effector cell proliferation or the transcription of TNF$\beta$. Upregulation of TNF$\beta$ was induced by incubating the antibody activated effector cells with BHV-1 infected D17 target cells for 10 h. Anti-TNF$\beta$ antibody partially abolished (13-44%) the direct effector cell-mediated antiviral cytotoxicity. However, this antibody neutralized 70 to 100% of antiviral activity of effector and target cell culture supernatants against BHV-1 infected D17 cells. The inhibition level of the antiviral activity by the antibody was dependent on the effector and target cell ratio. These results support the hypothesis that increased $56^ICK enzyme activity in effector cells transduces a signal critical for effector cell recognition of viral glycoproteins expressed on the target cells. Following target cell recognition, lytic cytokines known to participate in target cell killing were produced. A better understanding of the killing activity displayed by CD$4^+$T lymphocytes following surface receptor cross-linking will provide insight into the mechanisms of cytotoxic activity directed toward virally-infected cells.

• 생물정신의학
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• 제3권2호
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• pp.170-180
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• 1996

Lowered immune function in the senile dementia patients may be related to the abnormal metabolism of amyloid precursor protein(APP). To investigate the passibility of an abnormal metabolism of APP in lymphocytes and the possible role of APP in the activation of lymphocytes in senile dementia patients, immunohistochemical study of rat spleen and fluorescence activated cell sorter analysis(FACS) of human lymphocytes with the specific antigen far each lymphocyte and double fluorescent marker with antibody to APP were performed. After stimulating lymphocyte with phytohemagglutinin(PHA), APP mRNA and protein were extracted and quantitfied and the influence of ${\beta}$-amyloid protein($A{\beta}$) specific antibody on lymphocyte division was investigated. In spleen, the majority of cells showing $A{\beta}$ immunoreactivity was found in the T-sell dependent zone. FACS indicated that around 90% $CD_4(+)$ T-cells and 60% of $CD_8(+)$ T-sell were immunoreactive to $A{\beta}$ specific antibody(mAb 4G8). Northern blot analysis shows that lymphocyte APP mRNA was gradually increased to reach a maximum at 3 days after activation with lectin mitogen PHA. However, the $A{\beta}$ immunoreactivity an cell surface remained constant during stimulation with PHA, indicating that the release of APP(secreted farm of APP) might be increased. A very large increase in soluble APP secretion was observed in T-lymphocyte upon activation, but only law levels in the resting stale. Immunoblot was carried out an the protein obtained from cell lysate after stimulating lymphocyte by applying PHA to the cultured lymphocyte, and the result was that $A{\beta}$ band of immature farm under 116 KDa marker decreased as the duration of culture was increased after PHA stimulation. The monoclonal $A{\beta}$ specific(4G8) and polyclonal APP antibodies did not inhibit the [$^3H$]-thymidine uptake of mitogen-treated lymphocytes significantly, suggesting that mitogenesis can not be inhibited by specific $A{\beta}$ and polyclonal APP antibody. These results suggest that APP is expressed in T-cell and might be closely associated with the function of T-cells.

• 대한핵의학회지
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• 제27권1호
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• pp.118-122
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• 1993

전이성 분화갑상선암을 치료할 목적으로 사용된 $^{131}I$에 의한 염색체 손상여부를 검정하기 위하여 치료 전후의 자매염색분체교환(SCE)의 빈도 및 말초혈액의 백혈구수를 정상대조군과 환자군에서 조사하였다. 1) 정상인으로 설정한 대조군에 있어서의 자매염색분체교환 빈도수가 $3.8{\pm}0.4$인데 비하여 $^{131}I$ 투여전의 SCE 빈도가 $4.2{\pm}0.7$으로 차이를 보이지 않았다(p=0.17). 2) $^{131}I$ 투여후 2일째 SCE 빈도는 $7.9{\pm}0.8$으로 치료전에 비해 유의하게 증가하였으며 (p<0.001), 9일째에는 $6.4{\pm}0.6$으로 2일째에 비해서는 유의하게 감소하였으나(p=0.002), 치료전에 비해서는 아직도 유의하게 높았다 (P<0.001). 3) 같은 기간동안 백혈구의 수는 유의한 변화를 보이지 않았다. 결과적으로, $^{131}I$ 치료후 염색분체의 손상은 일어나며, 서서히 회복되나 1주일 이내에는 완전한 회복을 보이지 않음을 알 수 있었다.

• 대한핵의학회지
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• 제33권1호
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• pp.94-99
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• 1999

목적: 조사선량별로 인체 말초혈액 림프구에서 염색체 손상은 중기염색체 분석법으로, 그리고 세포고사는 annexin V를 이용한 유세포계측법으로 정량화하여 서로 비교해 봄으로써, 인체 말초혈액 림프구가 조사선량에 따라 세포사망에 이르는 과정을 밝혀보고자 하였다. 대상 및 방법: 염색체 이상을 동반하는 질환이 없는 건강한 자원자 10명(남자 6명, 여자 4명, 연령범위 $23{\sim}41세$)을 대상으로 하였으며, 이들로부터 혈액을 채혈하여 실험에 이용하였다. 채혈한 혈액으로부터 림프구를 분리하여 0.18, 2, 5, 10, 20. 25 Gy를 조사하였으며, 방사선을 조사하지 않은 림프구를 대조군으로 하였다. 방사선에 조사된 실험군과 대조군을 각각 중기염색체 분석법과 유세포계측법으로 검사하였으며, 중기염색체 분석법에서는 600개의 림프구로부터 불안정 염색체인 2중심염색체와 반지모양 염색체의 수를 계수하였고, 유세포 계측법에서는 세포막의 변화는 annexin V에 결합된 FITC의 섭취로, 그리고 세포핵의 변화는 PI의 섭취로 보았다. 실험군 간의 차이는 ANOVA 검사로, 그리고 두 검사법간의 상관관계는 Pearson의 상관검사로 알아보았다. 결과: 방사선을 조사하지 않은 세포군에서도 불안정 염색체가 $0.5{\pm}0.53$개 관찰되었으며, 실험군에서는 조사선량의 증가에 따라 불안정 염색체의 숫자가 유의하게 증가되었다(p<0.001). 조사선량의 증가에 따라 초기 세포고사는 증가하다가 다시 감소하는 양상을 보이나, 후기 세포고사는 조사선량에 따라 증가하는 양상을 보였다. 중기염색체 분석에서 보이는 불안정 염색체의 수와 Ydr, Qdr 값은 초기 세포고사와는 유의한 상관관계를 보이지 않고, 후기 세포고사와는 매우 유의한 상관관계를 보이고 있었다(p<0.01). 결론: 조사선량에 따른 불안정 염색체의 증가는 유세포계측법으로 검사한 세포핵의 변화와 밀접한 상관관계를 가지고 있었다. 반면 세포막의 변화는 염색체 손상과 관계가 없었으며, 조사선량의 증가에 비례하여 증가하지도 않았다.

• Clinical and Experimental Reproductive Medicine
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• 제27권4호
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• pp.405-415
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• 2000

Objective: The present study was undertaken to synthesize a human Y chromosome specific probe and to confirm the usefulness of the probe for fluorescence in situ hybridization (FISH) in various types of human cells. Methods: An approximately 400 bp DNA fragment of the DYZ1 sequences was synthesized by PCR using digoxigenin labeled dUTP (dig-PCR). The fidelity of probe was tested by FISH for cultured and uncultured human lymphocytes, amniocytes, chorionic villus cells, embryos, sperms, and germ cells of seminiferous tubule. Results: The human Y chromosome specific probe hybridized specifically to Y chromosome of the cells that had been tested. This probe assigned to the Yq12 region where the DYZ1 repetitive sequence is concentrated. Conclusion: We have identified a human Y chromosome specific probe that hybridized specifically to the Y chromosome by FISH for various types of uncultured as well as cultured cells. Therefore FISH technique using human Y chromosome specific probe should be useful for clinical application as a diagnostic tool for the detection of human Y chromosome.

• 대한소아치과학회지
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• 제10권1호
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• pp.25-33
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• 1983

With electron microscope, author studied on the pulp structure of human primary tooth in shedding stage. Non-carious human primary molar teeth were selected for this study. Using standard methods, specimens were sectioned and examined by light and electron microscope, The results were as follows; 1. In coronal pulp, odontoblasts were replaced by multinucleated odontoclasts, which contained a large number of mitochondria of varying shape and vacuoles in cytoplasm. Where odontoclasts were in contact with tooth surface, the characteristic ruffled border and clear zone were observed. 2. Fibrous tissue with plentiful collagen fibers and fibroblasts was observed adjacent to the dentin in the pulp. Fibroblast contained a number of mitochondria and well-developed rough-surfaced endoplasmic reticulum. 3. Inflammatory cells were observed in the pulp and active fibroblasts could be seen between inflammatory cells. In many cases, cervical epithelium proliferated toward absorbed area. 4. Inflammatory cells consisted of a number of lymphocytes, polymorphonuclear leukocytes, plasma cells and macrophages. Macrophage containing lysosomes in digestive state or phagocyting PMN could be seen. 5. In the primary molar of delayed root resorption, odontoblast layer, zone of Weil and cell-rich zone could be seen at roof of pulp chamber and odontoblast in this area cont과ained some lipid droplets.