• Development and Reproduction
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• v.14 no.4
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• pp.297-306
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• 2010

Tributyltin (TBT), an endocrine disrupting chemical, has been reported to decrease testicular function by causing apoptosis in the testis, but this mechanism is not fully understood. Thus, in this study we examined whether TBT induces adipogenesis of the Leydig cells to find out the correlation between adipogenesis and apoptosis in the testis. Three week old SD male rats were orally administrated with sesame oil, 1 mg/kg of TBT, or 10 mg/kg of TBT daily for 1 week and weighed after administration. The testes obtained on day 8 were weighed and stained with BODIPY and TUNEL kit. Using total RNA extracted from the isolated Leydig cells, adipogenesis and apoptosis-related genes were analyzed by real-time PCR. The testicular weights of the rats treated with 10 mg/kg TBT were significantly decreased compared to those in the control rats treated with sesame oil. As a result of BODIPY staining, the number of Leydig cells stained with BODIPY was increased in the rats treated with 10 mg/kg TBT compared with the control rats. Similar to BODIPY staining results, the TUNEL assay showed that the apoptosis of Leydig cells was increased in TBT treated rats. The results of the gene expression analysis in the Leydig cells showed that the expression of adipogenesis-related genes (PPAR${\gamma}$, aP2, Perilipin, CD36) and apoptosis-related genes (TNFRSF1A, TNFSF10) was increased after TBT administration. The present study demonstrates that TBT induces the expression of adipogenesis-related and apoptosis-related genes in the Leydig cells leading to adipogenesis and apoptosis in the testes. These results suggest that the dysfunction of Leydig cells by TBT exposure may cause a loss in testicular function.

• Radiation Oncology Journal
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• v.17 no.4
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• pp.299-306
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• 1999

Purpose : The human genetic disorder ataxia-telangiectasia (AT) is a multisystem disease characterized by extreme radiosensitivity. The recent identification of the gene mutated in AT, ATM, and the demonstration that it encodes a homologous domain of phosphatidylinositol 3-kinase (PI3-K), the catalytic subunit of an enzyme involved in transmitting signals from the cell surface to the nucleus, provide support for a role of this gene in signal transduction. Although ionizing radiation was known to induce c-fos transcription, nothing is known about how ATM or PKCI mediated signal transduction pathway modulates the c-fos gene transcription and gene expression. Here we have studied the effect of PKCI on radiation sensitivity and c-fos transcription in normal and AT cells. Materials and Methods: Normal (LM217) and AT (AT5BIVA) cells were transfected with PKCI expression plasmid and the overexpression and integration of PKCI was evaluated by northern blotting and polymerase chain reaction, respectively. 5 Gy of radiation was exposed to LM and AT cells transfected with PKCI expression plasmid and cells were harvested 48 hours after radiation and investigated apoptosis with TUNEL method. The c-fos transcription activity was studied by performing CAT assay of reporter gene after transfection of c-fos CAT plasmid into AT and LM cells. Results: Our results demonstrate for the first time a role of PKCI on the radiation sensitivity and c-fos expression in LM and AT cells. PKCI increased radiation induced apoptosis in LM cells but reduced apoptosis in AT cells. The basal c-fos transcription activity is 70 times lower in AT cells than that in LM cells. The c-fos transcription activity was repressed by overexpression of PKCI in LM cells but not in AT cells. After induction of c-fos by Ras protein, overexpression of PKCI repressed c-fos transcription in LM cells but not in AT cells Conclusion: Overexpression of PKCI increased radiation sensitivity and repressed c-fos transcription in LM cells but not in AT cells. The results may be a. reason of increased radiation sensitivity of AT cells. PKCI may be involved in an ionizing radiation induced signal transduction pathway responsible for radiation sensitivity and c-fos transcription. The data also provided evidence for novel transcriptional difference between LM and AT cells.

• Parasites, Hosts and Diseases
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• v.26 no.4
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• pp.229-238
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• 1988

In vitro culture of Toxoplasma gondii in HL-60 cells and cell-mediates immunity against Toxoplasma in dimethylsulfoxide(DMSO) -induced HL-60 cells, i.e., differentiation into granulocytes, were pursued. HL-60 calls were treated with various concentrations of DMSO, and 1.3%(v/v) for 3 day incubation was chosen as the optimal condition icy differentiation into granulocytes. The degree of differentiation was assayed in physiological and functional aspects in addition to morphological point. When treated with 1.3% DMSO for 3 days, HL-60 cells did not synthesiar DNA materials beyond background level, and showed active chemotactic response to chemotactic peptide, formal-methionyl-leucyl-phenylalanine(FMLP). Morphologically promyelocytes of high nuclearlcytoplasmic(NIC) ratio changed to granulocytes of relatively low WJC ratio. The relationships between HL-60 cells or DMSO-induced HL-60 cells and Toxoplasma were examined after stain with Giemsa and Buorescent dye (acridine orange). HL-60 cells did not show any sign of torso- plasmacidal activity but showed intracellular proliferation of Texoplasma to form rosette for 72 hr co-culture. In contrast, OMSO-induced HL-60 cells phagocytosed Toxoplasma within 1 hr, and performed a process of intracellular digestion of Toxoplasma thereafter. With the above results, it is suggested that phagosome-Iysosome fusion is one of the critical events for the parasitism by Toxoplasma or for susceptibility of host cells. The in vitro culture system of this study has offered a defined condition to study the protozoan parasite-host cell interactions.

• Journal of Plant Biology
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• v.37 no.2
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• pp.183-193
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• 1994

인삼(Panax ginseng C.A. Meyer)의 뇌두 캘러스로부터 분리한 원형질체와 배양된 원형질체의 미세구조 변화와 원형질막 표면을 투과 및 주사전자현미경으로 조사하였다. 분리 직후의 원형질체에서는 캘러스세포에서보다 작은 액포들이 많이 형성되어 있었다. 또한 활면소포체의 수가 증가하였으며 이들은 원형질막과 평행으로 배열하였다. 활면소포체는 가끔 세포질을 둘러싸서 세포질분리체(cytosegresome)를 형성하였고, 이 구조는 세포질을 분해시킨 후 액포로 변화하기도 하였다. 배양된 원형질체에서는 분리 직후의 원형질체와 비교하여 조면소포체, 딕티오좀, 리보좀, 미토콘드리아, proplastid 및 액포 등의 수가 현저하게 증가하였다. 딕티오좀으로부터 많은 소낭들이 형성되었고 이들은 세포질 전반에 걸쳐 존재하였다. 때로는 소낭들이 원형질막 바깥으로 돌출되어 돌기를 형성하기도 하였다. 배양된 원형질체의 표면에는 섬유소로 구성되어 있을 것으로 추정되는 섬유상 구조들이 형성되어 있었다.

• The Korean Journal of Zoology
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• v.38 no.2
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• pp.186-195
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• 1995

사람의 항 HLA 단일군 항체 생산의 선결 조건인 가장 효과적인 시험관내 면역 조건을 확립하기 위해, 사람의 혈중 임파구를 함원으로 하여 마우스의 대식세포, 흉선세포. 이들의 조건배지, 그리고 임파구 촉진인자 등을 포함한 14가지의 다양한 배양 조건에서 세포를 배양하였으며, 마우스의 비장세포와 사람의 혈중 임파구에서 각각 항체 생산을 유도하였다. 항체의 생성 여부는 면역효소법(I섬SA)으로 조사하였다. 마우스의 비장세포는 모든 조건에서 다량의 항체가 검출되었으며, 사람의 혈중 임파구 분화에는 마우스의 조건배지보다 PWM. LPS와 같은 임파구 촉진인자가 효과적임을 알 수 있었으며. 특히 allogenic MLC(Mixed Lymphocyte Culture)에 의해 임파구 분화유도에 유용한 물질이 생성됨을 알 수 있었다.

• The Korean Journal of Zoology
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• v.30 no.4
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• pp.325-340
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• 1987

근세포 융합에 있어서 당단백질의 역할을 세포내 glycosylation의 저해제인 tunicamycin을 이용하여 검토하였다. 근세포가 읖합하기 전 여러 시기에 tunicamycin을 0.04091m숙 농도로 처리하면 세포내 glycosylation과 근세포 융합은 크게 감소되지만, 단백질 합성률과 creatine kinase 활성은 별로 변하지 않는 점으로 미루어 보아 근세포 표면의 당단백질은 세포간의 recognition과 adhesion에 관여하는 것으로 추정할 수있었다. 따라서, 표지된 Con A 염색법을 써서 근세포 원형질막의 당단백질의 변화를 검토해 본 결과 tunicamycin을 처리한 경우 원형질막 당단백질의 대부분이 감소됨을 볼 수 있었으며, 아울러 근세포내 단백질의 분해속도는 증가하고 fibronectin은 감소하는 현상을 관찰할 수 있었다. 한편, fibronectin(20 ug/ml) 을 tunicamycin과 같이 처리한 경우에는 융합이 억제되지 않았다. 이상의 결과들은 tunicamycin이 근세포가 adhesion하는데 필요한 세포막표면의 fibronectin의 유용성을 감소시킴으로써 근세포의 융합을 억제할 가능성을 제시하는 것이다.

• Journal of Veterinary Clinics
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• v.15 no.1
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• pp.100-103
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• 1998

아까바네 바이러스로 인하여 유산된 태아의 뇌조직으로부터 '아까바네 바이러스 항원을 면역학적으로 검출하는 기법을 확립하였다. 아까바네 바이러스로 유산된 태아의 뇌는 조 직이 거의 손실되거나 유약하여 부검 즉시 포르말린 등에 보존하여야 하므로, 포르말린에 보존 된 뇌조직을 절편 하여 파라핀으로 포매된 조직표본으로부터 immunohistochemistry 방법으로 아까바네 바이러스 특이 항원을 검출하였다. 또한 이들 조직으로부터 직접 마우스 뇌내 접종과 조직배양내 바이러스 분리를 통하여 immunohistochemistry 법의 항원 검출 효율이 높음을 확 인하였다. 유산된 태아의 뇌조직에서 단크론 항체를 이용한 항원 검출 실험에서 세포의 세포질 내에서 아까바네 특이 항원이 검출되었고 hematoxylin-rosin 대조 염색으로 항원을 특이적으로 구분하여 진단할 수 있었다. 바이러스에 감염된 세포는 조직학적으로 변성이 심한 부위에서 다 수 관찰되었고 맥관계에 가까운 세포에서도 독립적으로 감염된 세포가 관찰되었다.

• Korean Journal of Food Science and Technology
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• v.47 no.1
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• pp.81-86
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• 2015

Intense pulsed light (IPL), a nonthermal technology, has attracted increasing interest as a food processing technology. However, its efficacy in inactivating microorganisms has not been evaluated thoroughly. In this study, we investigated the influence of IPL treatment on the inactivation of Escherichia coli O157:H7 depending on light intensity, treatment time, and pulse number. Increased light intensity from 500 V to 1,000 V, raised the inactivation rate at room temperature. At 1000 V, the cell numbers were reduced by 7.1 log cycles within 120 s. In addition, increased pulse number or decreased distance between the light source and sample surface also led to an increase in the inactivation rate. IPL exposure caused a significant increase in the absorption at 260 nm of the suspending agent used in our experiments. This indicates that IPL-treated cells were damaged, consequently releasing intracellular materials. The growth of IPL-irradiated cells were delayed by about 5 h. The degree of damage to the cells after IPL treatment was confimed by transmission electron microscopy.

• Journal of Acupuncture Research
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• v.20 no.2
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• pp.112-122
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• 2003

연구 배경 및 목적 : 봉독은 항염증 등의 효과를 가지고 있다고 알려져 있지만 세포 증식과 관련된 봉독의 효과에 대한 고찰을 위해 본 연구(硏究)에서는, 봉독(蜂毒) 약침(藥鍼) 자극(刺戟)이 아세트산 유발(誘發) 통각(痛覺) 과민증(過敏症)을 가진 쥐의 치상회(齒狀回)에서의 세포(細胞) 증식(增植)과 COX2 발현(發現)에 미치는 영향(影響)에 대해 알아보고자 하였다. 실험방법 : 대조군, 아세트산 처치군, 아세트산 0.1mg/kg 봉독 처치군, 아세트산 1mg/kg 봉독처치군(n=5 in each group)의 네 군으로 나누고 해당 군에 일벌 봉독(蜂毒)(Sigma Chemical Co., St. Louis, MO, USA)을 양측 족삼리 경혈(ST36)에 주입시키고 30분 후 아세트산(100% acetic acid 1% 용액의 0.5ml)을 복강내로 주입하여 복부 긴장 회수를 세었으며, BrdU 양성, COX 2 양성 세포수를 면역 화학 조직법을 수행하여 세어 보았다. 결과 : 아세트산 처치군에서는 대조군에 비해 5 bromo 2' deoxyuridine 양성 세포의 수는 감소(減少)되며, 치상회(齒狀回)에서의 COX 2의 발현(發現)은 증강(增强)되는 것으로 보여졌다. 봉독(蜂毒) 주입(注入)은 아세트산 유발(誘發) 복부(腹部) 긴장(緊張) 횟수와 치상회(齒狀回)에서의 COX2 발현(發現)을 억제(抑制)하여, 치상회(齒狀回)에서의 세포(細胞) 증식(增植)을 증가(增加)시켰다. 결론 : 이번 결과(結果)에서 보면, 치상회(齒狀回)에서의 COX 2의 발현(發現)은 세포(細胞) 증식(增植) 억제(抑制)와 관련(關聯)되며 봉독(蜂毒)은 COX 2 발현(發現) 억제(抑制)를 통해 치상회(齒狀回)에서의 새로운 세포(細胞) 형성(形成)을 증가(增加)시킨다는 것을 알 수 있다.

• Journal of the Korean Institute of Intelligent Systems
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• v.14 no.4
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• pp.411-417
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• 2004

Recently, instruments and systems related on biological technology have been enormously developed. Particularly, many researches for biological cell injection have been carried out. Usually, excessive contact force occurring when the end-effector and a biological cell contact might make a damage on the cell. Unfortunately, the excessive force could easily destroy the membrane and tissue of the cell. In order to overcome the problem, we proposed a new injection system for biological cell manipulation. The proposed injection system can measure the contact force between a pipette and a cell by using a force sensor. Also, we used vision technology to correctly guide the tip of the pipette to the cell. Consequently, the proposed injection system could safely manipulate the biological cells without any damage. This paper presents the introduction of our new injection system and design concepts of the new micro end-effector. Through a series of experiments the proposed injection system shows the possibility of application for precision biological cell manipulation such as DNA operation.