• Title/Summary/Keyword: Dead cells

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Granulocyte-macrophage colony stimulating factor protects dendritic cells from anticancer drug-induced apoptosis (수지상세포에서 GM-CSF의 항암제유도 세포사멸 방지효과에 관한 연구)

  • Joo, Hong-Gu
    • Korean Journal of Veterinary Research
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    • v.43 no.4
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    • pp.607-613
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    • 2003
  • Dendritic cells (DCs) play an essential role in a variety of immune reactions involving $CD4^+$ T cells and have been used to enhance tumor-specific immune responses. Immunosuppression in patients with cancer includes the downregulation of function and number of DCs. Although DCs have been studied, the apoptosis of Des induced by anticancer drugs for chemotherapy remains largely uncharacterized. This study demonstrated that GM-CSF protects DCs from 5-fluorouracil (5-FU) or mitomycin C-induced apoptosis. After 6 - 10 days culture, DCs were characterized by specific surface marker, CD11c and MHC class II. MTT assay revealed that GM-CSF significantly enhanced the viability of DCs treated with 5-FU or mitomycin C. The percentage of dead cells of DCs was determined by cell size using FACScan and GM-CSF was clearly effective. However, GM-CSF did not increase the expression of MHC class II on viable DCs gated, suggesting that GM-CSF may differentially regulate critical factors involved in the function of DCs. For the quantitative analysis of apoptosis, annexin V-FITC staining was performed. 5-FU induced the apoptosis of DCs and GM-CSF significantly protects DCs from 5-FU-induced apoptosis. Taken together, the results in this study that GM-CSF has an anti-apoptosis effect on DCs may provide patients with cancer with clinical benefits to overcome the immunosuppression induced by the decrease of number and functional insufficiency of DCs.

14-3-3-Associated Proteins in Helicobacter pylori-Infected Gastric Epithelial Cells (Helicobacterpylori에 감염된 위상피세포에서 14-3-3 결합 단백질의 변화)

  • Chung, Hae-Yun
    • The Korean Journal of Food And Nutrition
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    • v.24 no.2
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    • pp.258-267
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    • 2011
  • 14-3-3 is a highly conserved, ubiquitously expressed protein family. It associates with diverse cellular proteins through its specific phosphoserine/phosphothreonine-binding activity and thus contributes to the regulation of crucial cellular processes such as metabolism, signal transduction, cell-cycle control, apoptosis, protein trafficking, transcription and stress responses. This study aims to determine changes in levels of 14-3-3 isoforms and 14-3-3 - associated proteins in Helicobacter pylori(H. pylori)-infected gastric epithelial AGS cells. AGS cells were stimulated with H. pylori(NCTC 11637) at the ratio of 300:1(bacterium:cell). Western blot analysis revealed that 14-3-3 $\sigma$ was elevated at 3 hr after H. pylori treatment. Other isoforms were not significantly affected by H. pylori infection. Using immunoprecipitation to 14-3-3 $\sigma$, followed by proteomic analysis, we found that S phase kinase associated protein isoform 2 bound to 14-3-3 $\sigma$ has increased. In contrast, three proteins (DEAD-box polypeptide 3, heterogeneous nuclear ribonucleoprotein H2 and WD repeat-containing protein isoform 1) bound to 14-3-3 decreased by H. pylori infection. Our results suggest that 14-3-3 may play an important regulatory role in H. pylori-induced signal transduction in gastric epithelial cells.

Application of qDVC Method for Measuring Viable Cells in Lakes (호수 생태계에서 살아있는 세균을 측정하기 위한 qDVC 방법의 적용)

  • Kim, Mi-Ree;Seo, Eun-Young;Choi, Seung-Ik;Ahn, Tae-Seok
    • Korean Journal of Microbiology
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    • v.42 no.3
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    • pp.205-209
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    • 2006
  • For measuring the viable cells in lakes, quantitative direct viable count (qDVC) method is applied. In the qDVC process, the final concentration of glycine is fixed as 2%. For confirming the effectiveness of qDVC for enumerating the viable cells, the viable bacterial numbers were measured by plate count, CTC reduction method and qDVC method at 5 different lakes. Among these 3 methods, the bacterial numbers by qDVC is $2.4{\sim}6.0$ times higher than those by the other 2 methods. And by the qDVC method, the viable cells were easily discriminated from dead or dormant cells.

Trial for Drug Susceptibility Testing of Mycobacterium tuberculosis with Live and Dead Cell Differentiation (세포 염색 방법을 이용한 결핵균 감수성 검사법)

  • Ryu, Sung-Weon;Kim, Hyun-Ho;Bang, Mun-Nam;Park, Young-Kil;Park, Sue-Nie;Shim, Young-Soo;Kang, Seongman;Bai, Gill-Han
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.3
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    • pp.261-268
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    • 2004
  • Background : The resurgence of tuberculosis and outbreaks of multidrug resistant (MDR) tuberculosis have increased the emphasis for the development of new susceptibility testing of the Mycobacterium tuberculosis for the effective treatment and control of the disease. Conventional drug susceptibility testings, such as those using egg-based or agar-based media have some limits, such as the time required and difficulties in determining critical inhibitory concentrations, but these are still being used in many diagnostic laboratories because of no better lternatives, considering cost and accuracy. To overcome these limits, a rapid and simple method for new susceptibility testing, using live and dead assays, was applied for a bacterial cell viability assay to distinguish dead from live bacterial cells based on two-color fluorescence. Materials and Methods Strains : Forty strains were used in this study, 20 susceptible to all antituberculosis drugs and the other 20 resistant to the four first line antituberculosis drugs isoniazid, rifampicin, streptomycin and ethambutol. Antibiotics : The four antibiotics were dissolved in 7H9 broth to make the following solutions: $0.1{\mu}g\;isoniazid(INH)/m{\ell}$, $0.4{\mu}g\;rifampicin(RMP)/m{\ell}$, $4.0{\mu}g\;streptomycin(SM)/m{\ell}$ and $4.0{\mu}g\;ethambutol(EMB)/m{\ell}$. Results : Live and dead Mycobacterium tuberculosis cells fluoresced green and red with the acridin (Syto 9) and propidium treatments, respectively. These results are very well accorded with conventional drug susceptibility testing by proportional method on Lowensen-Jensen media (L-J) containing 4 drugs (INH, RMP, EMB and SM), showing a 93.7 % accordance rate in susceptible strains and 95% in resistant strains. Conclusion : The results of the drug susceptibility testing using the live and dead bacterial cell assay showed high accordance rates compared with the conventional proportion method on L-J. This finding suggests that the live and dead bacterial cell assay can be used as an alternative to conventional drug susceptibility testing for M. tuberculosis strains.

The Effect of Blueberry on ROS Accumulation and Cell Death in Human Normal Breast Epithelial(MCF10A) and Breast Cancer(MCF7) Cells (블루베리가 정상유선세포와 유방암세포의 ROS 축적과 세포사멸에 미치는 영향)

  • Lee, Se-Na;Kang, Keum-Jee
    • The Korean Journal of Food And Nutrition
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    • v.21 no.4
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    • pp.416-424
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    • 2008
  • In an effort to elucidate the differential actions of blueberry(BB) in both normal and cancer cells, we utilized human breast cell lines to assess the accumulation of radical oxygen species(ROS) and ROS-associated apoptosis in both human normal breast epithelial(MCF10A) and breast cancer(MCF7) cells. BB extract was added to the cultures at a final concentration of $20{\mu}g/m{\ell}$ for 0(control), 6, 12, and 24 hr intervals. The MCF10A cells evidenced no marked ROS accumulation in the presence of BB, whereas the MCF7 cells evidenced clear ROS accumulation upon BB treatment from 12 hours forward. The number of dying or dead cells did not increase in the BB-treated MCF10A cell groups, whereas that number increased profoundly from 12 hr forward. Furthermore, the expression levels of certain stress-related, and pro- and antiapoptotic gene products evidenced differential responses to BB treatment between the MCF10A and MCF7 cell groups. These results indicate that the components of BB extract differentiate cancer cells by not preventing ROS accumulation within cells and by inducing ROS-associated cell death in cancer cells. However, no marked ROS accumulation or induction of cell death was noted in the normal breast epithelial cells. The fact that BB extract exerted a differential effect on cancer cells opens further directions of research regarding the specific components that exert the differential BB-mediated effects in the selective prevention of normal cells and therapy for cancer tissues in the physiological body.

Expression of TASK-1 channel in mouse Leydig cells

  • Min Seok Woo;Eun-Jin Kim;Anjas Happy Prayoga;Yangmi Kim;Dawon Kang
    • Journal of Animal Reproduction and Biotechnology
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    • v.38 no.4
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    • pp.291-299
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    • 2023
  • Background: Leydig cells, crucial for testosterone production, express ion channels like ANO1 that influence hormone secretion. This study investigates the expression and role of the Tandem of P domains in a weak inward rectifying K+ channel-related Acid-Sensitive K+-1 (TASK-1) channel in these cells, exploring its impact on testicular function and steroidogenesis. Methods: TASK-1 expression in Leydig cells was confirmed using immunostaining, while RT-PCR and Western Blot (WB) validated its expression in the TM3 Leydig cell line. The effect of a TASK-1 channel blocker on cell viability was assessed through live/dead staining and MTT assays. Additionally, the blocker's effect on testosterone secretion was evaluated by measuring testosterone levels. Results: Immunohistochemical analysis revealed a predominant presence of TASK-1, along with c-Kit and ANO-1, in Leydig cells adjacent to seminiferous tubules and also in Sertoli and spermatogenic cells. Expression levels of TASK-1 mRNA and protein were significantly higher in TM3 Leydig cells compared to TM4 Sertoli cells. In addition, blocking TASK-1 in TM3 cells with ML365 induced cell death but did not affect LH-induced testosterone secretion. Conclusions: These findings suggest that TASK-1 in Leydig cells is crucial for their viability and proliferation, highlighting its potential importance in testicular physiology.

Survival Ability of Porcine Oocytes Frozen-Thawed by Open Pulled Straw Method (Open Pulled Straw 방법에 의해 동결-융해된 돼지난자의 생존능력)

  • 김세웅;박춘근;정희태;양부근;김정익
    • Journal of Embryo Transfer
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    • v.16 no.2
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    • pp.117-125
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    • 2001
  • Vitrification of oocytes has been applied recently fur pigs, but remains elusive. The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) on in vitro survival of porcine oocytes. When immature follicular oocytes frozen-thawed were cultured for in vitro maturation, maturation rates to metaphase-II stage were higher in oocytes with (25%) than without (15%) cumulus cells. After In vitro fertilization of oocytes frozen-thawed, the maturation rates were also significantly (P<0.05) higher in oocytes with (41%) that than without (17%) cumulus cells. However, the penetration rates were higher in oocytes without (19%) that than with (9%) cumulus. In another experiment, porcine oocytes matured in vitro were frozen and thawed for in vitro fertilization. The penetration rates were higher than in oocytes without (35%) that than with (26%) cumulus cells. However, the proportions of oocytes dead after in vitro fertilization were significantly (P<0.05) higher in oocytes with that than without cumulus cells. On the other hand, the rates of penetration and dead oocytes at 6 h after in vitro fertilization were not significant differences between oocytes with and without cumulus cells. However, the proportions of dead oocytes with (18%) and without (16%) cumulus cells were higher than in oocytes of control group (0%). These finding indicated the possible broader application for OPS, as they demonstrated that the maturation and fertilization in vitro by frozen-thawed oocytes may be accompained by cumulus cells and culture periods according to the requirements of the survival ability after freezing of mature and immature oocytes in pigs.

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Resource conservation using whole body autophagy: Self-digestion of shedded gut lining cells in the small intestine

  • Lee, Phil Jun;Cho, Namki;Yoo, Hee Min;Chang, Sun-Young;Ko, Hyun-Jeong;Kim, Hong Pyo
    • Korean Journal of Food Science and Technology
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    • v.52 no.3
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    • pp.244-248
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    • 2020
  • To retain valuable resources, organisms adopt several strategies including coprophagy. Cells covering the outer skin and internal digestive lumen are actively recycled to maintain their integrity. In present study, we suggested that the small intestine can consume dead cells in a manner similar to how it consumes protein from the diet. We examined the eluates from five segments of the mouse small intestine and cecum and 2 segments of the large intestine and small intestine tissue, and detected immunoreactivity with eukaryotic caveolin-1 and β-actin antibodies only in the cecum and 2 segments from the large intestine. Bacterial agitation of the mouse intestine with Shigella disrupted the architecture and absorptive function of the small intestine. Small intestine eluates were immunoreactive with murine caveolin-1 and contained heme as determined by dot blot analysis. We concluded that the body conserves resources in the small intestine by disposing of and recycling shedded cells.

A Numerical Study on the Effect of Battery-pack Shape of Electric Vehicle on the Forced Convection Around Battery Cells (전기자동차 배터리 팩 형상이 배터리 셀 주위의 강제대류에 미치는 영향에 대한 수치해석)

  • Kim, Kyo Hyeon;Kim, Tae Wan;Woo, Man Gyeong;Jeon, Byoung Jin;Choi, Hyoung Gwon
    • Journal of the Semiconductor & Display Technology
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    • v.16 no.1
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    • pp.16-21
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    • 2017
  • In this paper, the effect of battery-package shape of electric vehicle on the forced convection around a group of battery cells has been numerically investigated. Simulations for the two package shapes with straight/curved ducts have been conducted to examine the two design factors; the maximum temperature and the temperature deviation of a group of cells which influence the cell durability. The simulation of the conjugate heat transfer has been simplified by employing an equivalent thermal conductivity of cell that consists of various materials. It has been found that the maximum temperature and the temperature deviation of curved duct were lower than those of straight duct. Velocity fields have also been examined to describe the temperature distribution of a group of cells and the position of maximum temperature was found to be related to the dead zone of flow field.

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The effects of ascorbic acid on the morphology and cell proliferation of three-dimensional stem cell spheroids (아스코르브산의 3차원 줄기세포 배양체에 대한 증식 및 분화 효과 분석)

  • Lee, Hyunjin;Yeo, Seong-Il;Park, Jun-Beom
    • The Journal of the Korean dental association
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    • v.55 no.9
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    • pp.592-603
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    • 2017
  • Purpose: The effects of various concentrations of ascorbic acid on stem cell spheroids derived from intraoral areas are not known yet. Thus, the purpose of this study is to evaluate the effects of different concentrations of ascorbic acid on the morphology and cellular viability of stem cell spheroids derived from the gingival tissues. Materials and Methods: Stem cells were plated onto silicon elastomer-based concave microwells and grown in the presence of ascorbic acid at concentrations ranging from 0.003% to 0.3%. The morphology of the cells was viewed under an inverted microscope at day 1, 2, 3 and 5. Qualitative live/dead assay and quantitative cellular viability using Cell Counting Kit-8 were performed on day 2 and day 5. Results: Gingiva-derived stem cells formed spheroids irrespective of ascorbic acid concentration in silicon elastomer-based concave microwells. Increase in the diameter of spheroid were seen with higher concentrations of ascorbic acid. Higher cellular viability was seen in higher concentrations of ascorbic acid. Conclusion: Within the experimental setting, the application of ascorbic acid on stem-cell spheroids produced an increase in the size and higher viability with higher dosage. It can be suggested ascorbic acid be applied with stem cell spheroids for tissue engineering purposes.

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