• 제목/요약/키워드: Viability assessment

검색결과 178건 처리시간 0.023초

Cytotoxicity Assessment of Six Different Extracts of Abelia triflora leaves on A-549 Human Lung Adenocarcinoma Cells

  • Al-Taweel, Areej Mohammad;Perveen, Shagufta;Fawzy, Ghada Ahmed;Ibrahim, Taghreed Abdou;Khan, Afsar;Mehmood, Rashad
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권11호
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    • pp.4641-4645
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    • 2015
  • The present investigation was designed to assess the anticancer activity of six different leaf extracts (ethyl acetate, methanol, chloroform, petroleum ether, n-butanol, and water soluble) of Abelia triflora on A-549 human lung adenocarcinoma epithelial cells. A-549 cells were exposed to $10-1000{\mu}g/ml$ concentrations of the leaf extracts of A. triflorafor 24 h and then percentage cell viability was assessed by 3-(4,5-dimethylthiazol-2yl)-2,5-biphenyl tetrazolium bromide (MTT) assay. The results showed that leaf extracts of A. triflora significantly reduced the viability of A-549 cells in a concentration-dependent manner. Decrease was recorded as 31% with ethyl acetate, 36% with methanol, 46% with chloroform, 54% with petroleum ether, 62% with n-butanol, and 63% with water soluble extracts at $1000{\mu}g/ml$ each. Among the various plant extracts, ethyl acetate extract showed the highest decrease in the percentage cell viability, followed by methanol, chloroform, petroleum ether, n-butanol, and water soluble extracts. Our results demonstrated preliminary screening of anticancer activity of different soluble extracts of A. triflora extracts against A-549 cells, which can be further used for the development of a potential therapeutic anticancer agents.

아래쪽배벽동맥피판의 생존향상을 위한 VEGF와 COMP-angiopoietin-1 유전자 치료 (Improvement of the Inferior Epigastric Artery Flap Viability Using Adenovirus-mediated VEGF and COMP-angiopoietin-1)

  • 유은경;손대구;김형태;이인규;최태현;김준형;한기환
    • Archives of Plastic Surgery
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    • 제36권1호
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    • pp.1-10
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    • 2009
  • Purpose: Partial necrosis of skin flaps remains a substantial problem in reconstructive surgery. We investigated the potential use of an adenovirus vector encoding the VEGF, COMP-angiopoietin-1 gene in an attempt to promote the viability of the inferior epigastric artery flap in a rat model. Methods: Three by six cm lower abdominal transverse skin flaps, supplied only by the left inferior epigastric artery, were designed. After skin flap elevation, the adenovirus VEGF and adenovirus COMP-angiopoietin-1 were injected into the distal portion of the flap, which has a high tendency of developing flap ischemia. Control animals were injected with the same volume of normal saline. On 3, 7 and 14 days after the flap elevation, the flap survival and vascularization were assessed using Visitrak digital$^{(R)}$, CD31 immunohistochemistry in addition to evaluating the general histological characteristics. Results: There was a significant increase in the mean percentage of flap viability by 89.8%, 91.1% and 94.8% in flaps transfected with adenovirus VEGF, COMP-angiopoietin-1, coadministraion of VEGF and COMP-angiopoietin-1 at seven days, and by 95.6%, 94.8% and 96.3% at 14 days. Histological assessment revealed that there were more blood vessels formed after adenovirus with VEGF, COMP-angiopoietin-1 or VEGF plus COMP-angiopoietin-1 than with adenovirus Lac Z. Conclusion: The results of this study suggest that adenovirus-mediated VEGF, COMP-angiopoietin-1 gene therapy, promote therapeutic angiogenesis in patients that undergo reconstructive procedures.

Assessment of Sperm Characteristics in Fresh and Frozen Semen of Miniature-Pig

  • Lee S. H.;Kim T. S.;Cheong H. T.;Yang B. K.;Kim C. I.;Park C. K.
    • Reproductive and Developmental Biology
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    • 제28권4호
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    • pp.261-265
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    • 2004
  • The present study was conducted to assess sperm characteristics in miniature-pig. The semen samples were transported to the laboratory at 17℃ within 3 hours after collection. The extended semen was stored at 17℃, and sperm quality was evaluated at 0, 1, 3, 5 and 7 days after storage. The semen volume of miniature-pig (62±22㎖) was significantly (p<0.05) lower than that of Duroc (155±25㎖) and Yorkshire (154±23㎖). Significant differences were also observed in sperm concentrations. During 3 days of storage, sperm viability did not differ among miniature-pig, Duroc and Yorkshire. However, the viability was significantly (p<0.05) lower in miniature-pig than in Duroc and Yorkshire semen after Day 3 of storage. In abnormality, acrosome intactness and intensity, there were no differences among miniature-pig, Duroc and Yorkshire semen. On the other hand, the viability of frozen-thawed sperm in miniature-pig was significantly (p<0.05) lower than in that of Duroc and Yorkshire. This study also examined CTC patterns in frozen-thawed spermatozoa. The rates of AR pattern were higher in miniature-pig than in Duroc and Yorkshire. However, no difference was found in F, B and AR patterns. The results of present study suggest that further research is necessary to develop of semen extender and freezing methods to improve sperm quality in miniature-pig.

Development of Semen Extenders by Assessment of Sperm Viability in Miniature-Pig Semen

  • Lee S. H.;Cheong H. T.;Yang B. K.;Park C. K.
    • Reproductive and Developmental Biology
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    • 제29권4호
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    • pp.247-252
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    • 2005
  • The purpose of this study was to assess sperm quality during in vitro storage of miniature-pig semen in order to determine which extender should be used and how extender can be diluted for in vitro storage of miniature-pig semen. Freshly ejaculated miniature-pig's semen was diluted with same volumes of Beltsville Thawing Solution (BTS), Androhep, Modena, Mulberry III and modified-Modena extenders. Sperm quality was evaluated by examining viability, motility, abnormality, acrosome intactness, intensity and capacitation status by chlorotetracycline (CTC) staining. Sperm motility decreased with storage period prolonged and differences among BTS, Androhep, Modena and Mulberry III were apparent On Day 1, approximately 80% of the sperm were motile, but motility decreased to $40\%$ at Day 7. During the 7 days of storage, sperm survival in modified-Modena B extender was higher than another extenders. However, it was not differ significantly among other extenders. The percentage of F and B patterns were not differ significantly among the extenders. However, F pattern in modified-Modena B extender was slightly higher until 3 days of storage than that of Modena extender, modified-Modena A extender and modified-Modena C extender. The percentage of AR patterns in modified-Modena B extender was slightly lower, but did not differ significantly among other extenders. The results of present study suggest that modified-Modena B was effective as new extender for in vitro storage of miniature-pig semen.

Cardamonin Inhibited IL-1β Induced Injury by Inhibition of NLRP3 Inflammasome via Activating Nrf2/NQO-1 Signaling Pathway in Chondrocyte

  • Jiang, Jianqing;Cai, Mingsong
    • Journal of Microbiology and Biotechnology
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    • 제31권6호
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    • pp.794-802
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    • 2021
  • In this study we investigated the role and mechanism of cardamonin on IL-1β induced injury in OA. CHON-001 cells were treated with cardamonin and IL-1β and transfected with silencing nuclear factor erythroid 2-related factor 2 (siNrf2). Cell viability was detected by Cell Counting Kit-8 assay and flow cytometer assay was utilized for cell apoptosis assessment. IL-6, IL-8, TNF-α and Nrf2 mRNA expression was tested by qRT-PCR. Western blot was employed to evaluate MMP-3, MMP-13, Collagen II, Nrf2, NQO-1, NLRP3, Caspase 1 and apoptosis-associated speck-like protein containing a caspase-1 recruitment domain (ASC) protein levels. In CHON-001 cells, IL-1β suppressed cell viability and Collagen II level while promoting cell apoptosis and expression of pro-inflammatory cytokines (IL-6, IL-8, TNF-α), MMPs (MMP-3, MMP-13), NQO-1, and NLRP3 inflammasome (NLRP3, Caspase 1 and ASC), with no significant influence on Nrf2. Cardamonin reversed the effect of IL-1β on cell viability, cell apoptosis, pro-inflammatory cytokines, MMPs, Collagen II, and NLRP3 inflammasome levels. In addition, cardamonin advanced Nrf2 and NQO-1 expression of CHON-001 cells. SiNrf2 reversed the function of cardamonin on IL-1β-induced cell apoptosis and expression of pro-inflammatory cytokines, Nrf2, NQO-1, and NLRP3 inflammasome in chondrocytes. Taken together Cardamonin inhibited IL-1β induced injury by inhibition of NLRP3 inflammasome via activating Nrf2/NQO1 signaling pathway in chondrocyte.

Effects of ascorbic acid augmented albumin platelet-rich fibrin on the wound healing activity of human gingival fibroblasts: an in vitro trial

  • Manjiri Kulkarni;Sowmya NK;Gayathri GV;Triveni MG
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제50권4호
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    • pp.206-215
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    • 2024
  • Objectives: The current in vitro study aimed to assess the effects of ascorbic acid augmented albumin platelet-rich fibrin (AA Alb-PRF) on the wound healing activity of human gingival fibroblasts (HGFs) purported to be a regenerative biomaterial in surgical procedures. Materials and Methods: All assays were performed on three HGF groups, group I: complete media; group II: Alb-PRF, and group III: AA Alb-PRF. Alb-PRF was prepared following the protocol by Fujioka-Kobayashi et al. (2021). For preparation of AA Alb-PRF, 2,500 ㎍ AA was added to the blood pre-centrifugation. All groups were subjected to 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to estimate cell viability and proliferation, scratch assay for migration (0, 4, 12, and 24 hours) and transwell migration assay for chemotactic migration assessment (24 hours). Outcome variables were optical density (OD) for MTT assay, percentage of wound closure in scratch assay, and number of migrated cells in transwell migration assay. One-way ANOVA for MTT and transwell migration assays and two-way ANOVA for scratch assay with Bonferroni correction were performed with significance set at P<0.05. Results: Cell viability and proliferation (OD: 0.684±0.003 and proliferation: 28%) and wound closure (49.92%±1.62% at 4 hours and 61.39%±0.88% at 12 hours) were significantly higher in group III, while group II demonstrated the maximum number of HGFs migrating across the transwell membrane (9.25±2.49) with P<0.05. Conclusion: HGFs demonstrated a significant increase in viability and proliferation along with rapid wound closure in the presence AA Alb-PRF compared to Alb-PRF alone, indicating additional beneficial effects of AA. Thus, AA Alb-PRF potentiates the wound healing activity of HGFs and could be employed in oral, maxillofacial, and periodontal surgeries as a regenerative biomaterial.

Protective Effect of Aminoglycosides and Their Combinations Against 2-Chloroethylethyl Sulfide Exposure

  • Kim, Yun-Bae;Hur, Gyeung-Haeng;Choi, Dae-Sung;Shin, SungHo;Cha, Seung-Hee;Park,Yong-Keun;Sok, Dai-Eun
    • Toxicological Research
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    • 제13권1_2호
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    • pp.61-69
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    • 1997
  • Exposure of splenocytes to 2-chloroethylethyl sulfide (CEES) resulted in the cell death, and the cytotoxicity of CEES was prevented by inhibitors of lysosomal hydrolases. Therefore, it has been postulated that the cytotoxicity of CEES may be partially due to the lysosomal labilization. This study, based on this mechanism, was undertaken to determine whether aminoglycoside antibiotics as inhibitors of lysosomal phospholipases and their combinations with other lysosome stabilizers can be useful as a treatment to reduce the CEES toxicity in mice. 2-Chloroethylethyl sulfide (20 mg/kg body weight) was injected ip into female ICR mice, and candidate compounds were administered ip before or after the CEES challenge. Kanamycin (40 mg/kg body weight) as effective as deferoxamine (100 mg/kg body weight) enhanced the survival rate after 5 days of intoxication from 10% of control to 50 - 60%. The most effective was found to be the combination of kanamycin, cycloheximide, deferoxamine and dextrose showing an almost full protection against 2LD50 of CEES. Consistent with the protection of the CEES toxicity, the decrease of body weight in mice intoxicated with CEES was effectively prevented by kanamycin or its combinations. It is suggested that kanamycin or its combination (kanamycin, cycloheximide, deferoxamine and dextrose) would be one of effective antidotes against the CEES poisoning in mice.

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격자형 토지적성평가방법의 개선방안 연구 (An Improved Method in Grid-based Land Suitability Assessment)

  • 김학열;안재성
    • Spatial Information Research
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    • 제13권1호
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    • pp.1-17
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    • 2005
  • 현재 시행되고 있는 토지적성평가체계는 관리지역의 보전 및 이용가능성의 정도에 따라 필지별로 생산$\cdot$보전$\cdot$계획관리지역으로 세분화하도록 규정하고 있다 본 연구는 현행 필지별 토지적성평가체계의 문제점을 분석하고 이에 대한 개선점을 도출하여 합리적인 평가방법을 제시하는 것을 목적으로 한다. GIS를 활용하여 사례지역을 대상으로 민감도 분석을 수행하여, 보다 상세한 필지정보를 획득할 수 있는 10m 해상도의 격자형 평가방법을 대안으로 제시하였다. 또한 단 하나의 기개발지 및 공공시설만을 선택하여 개발적성에 반영하는 현 평가방법의 대안으로써, 해당지역의 기개발지 및 공공시설의 분포를 고려하여 두개이상의 기개발지 및 공공시설을 선정하여 평가하도록 제안하였다. 마지막으로 개별 필지의 개발적성산출에 영향을 미칠 수 있는 기개발지 및 공공시설의 특성을 반영하는 방법으로, 중력 모형의 매력도 변수를 합리적으로 조정하여 보다 과학적인 평가모형을 제시하였다.

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EID3 Promotes Glioma Cell Proliferation and Survival by Inactivating AMPKα1

  • Xiang, Yaoxian;Zhu, Lei;He, Zijian;Xu, Lei;Mao, Yuhang;Jiang, Junjian;Xu, Jianguang
    • Journal of Korean Neurosurgical Society
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    • 제65권6호
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    • pp.790-800
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    • 2022
  • Objective : EID3 (EP300-interacting inhibitor of differentiation) was identified as a novel member of EID family and plays a pivotal role in colorectal cancer development. However, its role in glioma remained elusive. In current study, we identified EID3 as a novel oncogenic molecule in human glioma and is critical for glioma cell survival, proliferation and invasion. Methods : A total of five patients with glioma were recruited in present study and fresh glioma samples were removed from patients. Four weeks old male non-obese diabetic severe combined immune deficiency (NOD/SCID) mice were used as transplant recipient models. The subcutaneous tumor size was calculated and recorded every week with vernier caliper. EID3 and AMP-activated protein kinase α1 (AMPKα1) expression levels were confirmed by real-time polymerase chain reaction and Western blot assays. Colony formation assays were performed to evaluate cell proliferation. Methyl thiazolyl tetrazolium (MTT) assays were performed for cell viability assessment. Trypan blue staining approach was applied for cell death assessment. Cell Apoptosis DNA ELISA Detection Kit was used for apoptosis assessment. Results : EID3 was preferentially expressed in glioma tissues/cells, while undetectable in astrocytes, neuronal cells, or normal brain tissues. EID3 knocking down significantly hindered glioma cell proliferation and invasion, as well as induced reduction of cell viability, apoptosis and cell death. EID3 knocking down also greatly inhibited tumor growth in SCID mice. Knocking down of AMPKα1 could effectively rescue glioma cells from apoptosis and cell death caused by EID3 absence, indicating that AMPKα1 acted as a key downstream regulator of EID3 and mediated suppression effects caused by EID3 knocking down inhibition. These findings were confirmed in glioma cells generated patient-derived xenograft models. AMPKα1 protein levels were affected by MG132 treatment in glioma, which suggested EID3 might down regulate AMPKα1 through protein degradation. Conclusion : Collectively, our study demonstrated that EID3 promoted glioma cell proliferation and survival by inhibiting AMPKα1 expression. Targeting EID3 might represent a promising strategy for treating glioma.

형광단백질 발현 유전자변형 누에(Bombyx mori )의 환경위해성 평가연구 (The Study of Environmental Risk Assessment for Fluorescent Genetically Modified Silkworms)

  • 김현정;정철의;구태원;이훈복
    • 한국응용곤충학회지
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    • 제53권3호
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    • pp.199-207
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    • 2014
  • 곤충(초파리, 모기, 누에)은 해충방제, 유용물질생산, 의학연구 등을 위해 유전자변형 곤충으로 개발되어 왔지만, 아직까지 국내에서 유전자변형 곤충에 대한 환경위해성 평가 등이 거의 실시 되지 못하고 있다. 따라서, 본 연구에서 유전자변형 누에의 환경위해성 평가를 3가지 항목(이동성, 생존능력, 산란 및 부화율)으로 진행하였다. 첫째, 기본 사육 절차에서의 탈출가능성(이동성), 둘째, 사육환경으로부터 탈출시 생존 가능성(8개의 극한 환경조건; 고온, 저온, 건조, 습함, 먹이의 유무), 셋째, 비유전자변형 누에♀ ${\times}$ 비유전자변형 누에♂, 비유전자변형 누에♀ ${\times}$ 유전자변형 누에♂, 유전자변형 누에♀ ${\times}$ 비유전자변형 누에♂, 유전자변형 누에♀ ${\times}$ 유전자변형 누에♂으로부터 나온 산란 및 부화율을 비교 하였다. 유전자변형 누에와 비유전자변형 누에의 이동성은 통계적으로 차이가 없었으며, 산란 및 부화율 또한 통계적 차이가 없었다. 다만, 비유전자 변형 암수쌍에서 산란된 알의 부화율보다 유전자변형 누에♀와 비유전자변형 누에♂에서 산란된 알의 부화율이 통계적으로 낮은 결과를 보였다. 극한환경에서의 생존율 실험에서 상대적으로 유전자변형 누에가 비유전자변형 누에보다 생존율이 낮았으며, 특히 고온조건의 환경에서 통계적으로 생존율이 낮은 결과를 보였다. 저온 조건의 경우 누에 유충의 동면으로 인해 실험결과를 명확하게 얻을 수 없었다. 유전자변형 누에와 비 유전자변형 누에가 일부의 차이를 보였으나 모든 실험에서 유전자변형 누에의 값이 비유전자변형 누에보다 낮게 나타났으며, 결과적으로 이번 연구에서는 유전자변형 누에의 위해성은 비유전자변형 누에보다 적었다.