• IMMUNE NETWORK
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• 제11권6호
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• pp.383-389
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• 2011

Background: EY-6 is one of the newly synthesized indoledione derivatives to induce tumor cell-specific cell death. In this study, we investigated the mechanism of immunological death induced by EY-6 at mouse colon cancer cell as well as at the normal immune cell represented by dendritic cell. Methods: C57BL/6 mouse syngeneic colon cancer cell MC38 was treated with EY-6, and analyzed by MTT for viability test, flow cytometry for confirming surface expressing molecules and ELISA for detection of cytokine secretion. Normal myeloid-dendritic cell (DC) was ex vivo cultured from bone marrow hematopoietic stem cells of C57BL/6 mice with GM-CSF and IL-4 to analyze the DC uptake of dead tumor cells and to observe the effect of EY-6 on the normal DC. Results: EY-6 killed the MC38 tumor cells in a dose dependent manner (25, 50 and $100{\mu}M$) with carleticulin induction. And EY-6 induced the secretion of IFN-${\gamma}$ but not of TNF-${\alpha}$ from the MC38 tumor cells. EY-6 did not kill the ex-vivo cultured DCs at the dose killing tumor cells and did slightly but not significantly induced the DC maturation. The OVA-specific cross-presentation ability of DC was not induced by chemical treatment (both MHC II and MHC I-restricted antigen presentation). Conclusion: Data indicate that the EY-6 induced tumor cell specific and immunological cell death by modulation of tumor cell phenotype and cytokine secretion favoring induction of specific immunity eliminating tumor cells.

• 대한약침학회지
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• 제25권4호
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• pp.390-395
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• 2022

Objectives: Sweet bee venom (sBV) is purified from Apis mellifera, containing a high level of melittin-its main component. It has been used as a therapeutic agent for pain relief and anti-inflammation, as well as for treating neuronal abnormalities. Recently, there have been studies on the therapeutic application of sBV for anticancer treatment. In the present study, we investigated the pharmacological effect of sBV treatment in A549 human lung cancer cells. Methods: We used microscopic analysis to observe the morphological changes in A549 cells after sBV treatment. The MTT assay was used to examine the cytotoxic effect after dose-dependent sBV treatment. Molecular changes in sBV were evaluated by the expression of apoptosis marker proteins using western blot analysis. Results: Microscopic analysis suggested that the growth inhibitory effect occurred in a dose-dependent manner; however, cell lysis occurred at a concentration over 20 ㎍/mL of sBV. The MTT assay indicated that sBV treatment exhibited a growth inhibitory effect at a concentration over 5 ㎍/mL. On fluorescence activated cell sorting analysis, G0 dead cells were observed after G1 arrest at treatment concentrations up to 10 ㎍/mL. However, rapid cell rupture was observed at a concentration of 20 ㎍/mL. Western blot analysis demonstrated that sBV treatment modulated the expression of multiple cell death-related proteins, including cleaved-PARP, cleaved-caspase 9, p53, Bcl2, and Bax. Conclusion: sBV induced cell death in A549 human lung cancer cells at a pharmacological concentration, albeit causing hemolytic cell death at a high concentration.

• 공업화학
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• 제30권6호
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• pp.687-693
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• 2019

수소전기차와 발전을 시작으로 수소연료전지 시장이 성장하면서 연료전지와 수소의 수요가 증가하고 있으므로, 조기 상용화와 시장 활성화를 위하여 연료전지의 내구성과 연료 이용효율에 관한 연구가 진행되어야 한다. 본 연구에서는 연료전지의 성능과 연료 이용효율을 최적화하기 위하여 양극 닫힌계의 운전조건에 대한 연구를 수행하였다. 부하 전류에 대한 배출 조건과 수소 공급 압력이 고분자전해질 연료전지의 성능에 미치는 영향에 대하여 평가하였고, 전해질막 두께에 대한 물의 역확산 영향을 분석하였다. 양극 닫힌계에서 수소극에 쌓인 물은 연료전지 전압이 3% 감소한 경우에 솔레노이드 밸브를 열어 배출하였다. 수소 공급 압력은 0.1~0.5 bar, 배출 시간은 0.1~1 s까지 변화시키면서 실험을 수행하였다. NR 211 (25.4 um) 전해질막의 경우 0.1 bar의 수소 공급 압력과 0.1 s 배출 시간 조건에서 수소 이용효율 98.9%의 가장 높은 연료 이용효율을 보였지만 잦은 flooding으로 인하여 장시간 운전 시 연료전지의 성능이 감소하였다. 이에 반해 NR 212 (50.8 um)의 전해질막에서 생성된 물과 질소의 역확산 속도를 늦추어 배출 간격을 늘리고 연료 이용효율을 높일 수 있었다.

• 한국방사선학회논문지
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• 제16권6호
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• pp.779-786
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• 2022

본 연구는 천연 방사선 방호제로 단풍마 추출물이 수컷 쥐의 전립선과 심장에 미치는 영향을 관찰하기 위한 것이다. 단풍마는 남성 질환과 심장질환을 개선하는 것으로 알려져 있다. 본 연구에서는 감마선 10 Gy를 수컷 흰 쥐(Sprague Dawley Rat)의 전신에 조사하여 단풍마 추출물의 방사선 방호 효과를 확인하였다. 조사 후, 1일, 7일, 21일 차의 혈액학적 변화, 7일 차의 항산화 효소(SOD) 활성도 및 조직 변화를 관찰하였다. 96 마리의 수컷 SD Rat을 대조군(NC Group), 단풍마 추출물 투여군(DQ Group), 방사선 조사군(IR Group), 단풍마 추출물 투여 후 방사선 조사군(DQ+IR Group)의 4개 군으로 분류하였다. DQ+IR Group은 IR Group보다 더 높은 림프구, 혈소판 수치를 보였다. SOD 활성도도 DQ+IR Group은 IR Group보다 높은 수치를 보였다. NC Group과 DQ Group에서는 전립선 낭 내의 세포 수가 양호하고 전립선 낭 간의 간격이 상대적으로 좁았다. 그러나 IR Group에서는 사멸한 세포 수가 많이 증가하고 간격이 넓어짐을 확인할 수 있었다. DQ+IR Group은 IR Group과 유사하게 전립선 낭 간의 간격이 증가했지만 사멸한 세포 수가 많이 감소함을 확인할 수 있었다. NC Group과 DQ Group은 심혈관과 심근이 뚜렷하게 분리되어 있고 세포의 상태가 양호하다. 그러나 IR Group에서는 사멸한 세포의 수가 많았고 심혈관과 심근 경계의 파괴를 확인할 수 있었다. DQ+IR Group은 IR Group과 유사하게 사멸한 세포의 수가 많았고 심혈관과 심근의 경계가 넓어졌지만 파괴되지는 않았다. 따라서 단풍마 추출물은 전립선 및 심혈관에 대한 방사선 방호효과가 있다고 사료된다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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• pp.110-117
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• 2000

Relationship between monoclonal antibody (MAb) productivity and growth rate, and effects of high cell density on MAb production rate increased with increasing specifis growth rate in both suspended and immobilized continuous cultures indicate a positively growth-associated relationship between MAb productivity and growth rate. moreover, the specific production rate was higher in the immobilized cell culture than that in suspended one at all dilution rates. In order to clarify these phenomana, MAb mPNA experession and cell cycle distribution were investigated in bacth cultures with immobilized cells and suspended cells. RT-PCR was used for observation of MAb mRNA expression and a two-color bromodeoxyuridine (BrdU)/propidium iodide (PI) flow cytometry method for determination of cell cycle distribution. The results revealed that MAb nRNA expression until dead phase, which was longer than in suspended cell. The cell cycle distribution patterns were observed almost the same for both immobilized and suspended cells. Such results may imply that a high cell density state has positive influence on the mRNA expression and on growth-associated Mab productivity of T0405 cells.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.918-922
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• 2001

In a previous study, we have isolated a number of lactobacilli from Korean women, and one of them (KLB46) was identified as Lactobacillus crispatus by 16S rRNA gene sequencing. For the ecological treatment of bacterial vaginosis (BV) cell suspension of L. crispatus KLB46 was instillated into BV patients. L. crispatus KLB46 was found to persist for several days in cell suspension with no nutrients. In this study, in order to assess the influence of starvation on physiological activity, we compared the viability and culturability of KLB46 following suspension in various buffer solutions. A pair of in situ fluorescent dye was used to assess viability (i.e. membrane integrity) and the culturability was examined by plate count assay. A rapid epifluorescence staining method using the LIVE/DEAD Bacterial Viability Kit $(BacLight^{TM})$ was applied to estimate both viable and total counts of bacteria in cell suspension. $BacLight^{TM}$ is composed of two nucleic acid-binding stains ($SYTO\;9^{TM}$ and propidium iodide). $SYTO\;9^{TM}$ penetrates all bacterial membranes and stains the cells green while propidium iodide only penetrates cells with damaged membranes, therefore the combination of the two stains produces red fluorescing cells. Optimal staining conditions for $BacLight^{TM}$ were found to be with 0.0835M $SYTO\;9^{TM}$ and 0.05M propidium iodide for 15 min incubation at room temperature in dark. When cells were microscopically examined during 140 hours of starvation, the culturability decreased markedly while the viability remained relatively constant, which suggests that large fraction of KLB46 cells became viable but non-culturable (VBNC) upon starvation.

• 동아시아식생활학회지
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• 제20권1호
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• pp.30-36
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• 2010

This study was conducted to investigate the effects of blueberry extract on cell death, ROS and gene expression patterns associated with the anti-cancer activity in human breast cancer MCF7 cells. To accomplish this, 20 mg/mL concentration of blueberry extract was added to the cell culture for 0, 6, 12, 24 or 48 h, after which the effects were evaluated by various analyses. MTT assay showed that the cellular activities decreased rapidly during the first 12 h of treatment. During this period, dual staining with Hoechst33322 and propidium iodide also produced a similar trend in which the dead or dying cells increased sharply. Furthermore, evaluation of BrdU incorporation as an index for cell proliferation revealed a marked decrease during the first 12 h of treatment, suggesting that anticancer activity involves the inhibition of cell proliferation and induces cell death. ROS also increased according to the duration of the treatment, indicating intracellular accumulation is associated with the cell death. RT-PCR analysis revealed significant decreases in anti-apoptotic (Bax) and increases in pro-apoptotic gene expressions (Bci-2, caspase- 3, and 9) (p<0.05). Taken these together, blueberry extract induces ROS accumulation in MCF7 cells, causing inhibition of cell proliferation and eventually leading to cell death. This cell death was associated with apoptotic gene expression in blueberry-treated cells for up to 24 h.

• 한국동물생명공학회지
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• 제39권2호
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• pp.145-152
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• 2024

Background: Despite its anticancer activity, cisplatin exhibits severe testicular toxicity when used in chemotherapy. Owing to its wide application in cancer therapy, the reduction of damage to normal tissue is of imminent clinical need. In this study, we evaluated the effects of catechin hydrate, a natural flavon-3-ol phytochemical, on cisplatin-induced testicular injury. Methods: Type 2 mouse spermatogonia (GC-1 spg cells) were treated with 0-100 μM catechin and cisplatin. Cell survival was estimated using a cell proliferation assay and Ki-67 immunostaining. Apoptosis was assessed via flow cytometry with the Dead Cell Apoptosis assay. To determine the antioxidant effects of catechin hydrate, Nrf2 expression was measured using qPCR and CellROX staining. The anti-inflammatory effects were evaluated by analyzing the gene and protein expression levels of iNOS and COX2 using qPCR and immunoblotting. Results: The 100 μM catechin hydrate treatment did not affect healthy GC-1 spg cells but, prevented cisplatin-induced GC-1 spg cell death via the regulation of anti-oxidants and inflammation-related molecules. In addition, the number of apoptotic cells, cleaved-caspase 3 level, and BAX gene expression levels were significantly reduced by catechin hydrate treatment in a cisplatin-induced GC-1 spg cell death model. In addition, antioxidant and anti-inflammatory marker genes, including Nrf2, iNOS, and COX2 were significantly downregulated by catechin hydrate treatment in cisplatintreated GC-1 cells. Conclusions: Our study contributes to the opportunity to reintroduce cisplatin into systemic anticancer treatment, with reduced testicular toxicity and restored fertility.

• Mycobiology
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• 제33권1호
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• pp.1-6
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• 2005

Morphological characteristics of hyphal interaction between Grifola umbellata (Pers. Ex Fr.) Pilat and its companion fungus which related to sclerotia formation from hyphae were investigated by external observations, light microscopy and transmission electron microscopy (TEM). External observations showed that a dense antagonism line was formed by both G. umbellata and companion fungus after their hyphae contacted each other in dual culture. Many hyphal strands emerged on the colony of G. umbellata and differentiated to sclerotia from where hyphal strands crossed. Light microscope observations revealed the process of antagonism line formation. Mature antagonism with structural differentiation, was composed of three main layers: the rind, the rind underlayer and the hypha layer. TEM observations showed that after colonies hyphal contact, a series of reactions always occurred in both G. umbellata and companion fungus. Cells in the center of antagonism line were dead. Cells of G. umbellata adjacent to the antagonism line were usually large and hollow, with unilateral thickened wall, whereas those of companion fungus were empty, with thin or thick wall. Both hyphal interaction at the antagonism line may be one of the main reasons for sclerotia of G. umbellata differentiation from hypha.

• 한국전산유체공학회지
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• 제15권1호
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• pp.31-36
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• 2010

In the present study, computations for flow fields inside the CDI unit cells with electrodes and spacers have been made to evaluate their performance. Three types of unit cells that include a planar type, a serpentine channel type, and a spiral wound type were considered and their flow characteristics were compared. From the computational results, it is found that the serpentine channel type has a large flow resistance and can not guarantee the outflow flux for industrial applications. On the other hand, the planar type can sustain a large enough outflow flux but it's efficiency is low for the electrode-use because of the non-uniform velocity distribution inside the cell and dead zones in every corner. Finally, The spiral wound type has not only a large outflow flux as much as the planar type has, but also a high efficiency for the electrode-use because of uniform velocity distribution. From this comparison, we can expect that the spiral wound type of CDI unit cell would have a high performance deionization capability.