• Title/Summary/Keyword: viability assessment

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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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    • v.16 no.11
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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.

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

  • Yoo, Eun Kyung;Son, Daegu;Kim, Hyung Tae;Lee, In Kyu;Choi, Taehyun;Kim, Junhyung;Han, Kihwan
    • Archives of Plastic Surgery
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    • v.36 no.1
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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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    • v.28 no.4
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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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    • v.29 no.4
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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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    • v.31 no.6
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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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    • v.50 no.4
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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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    • v.13 no.1_2
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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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An Improved Method in Grid-based Land Suitability Assessment (격자형 토지적성평가방법의 개선방안 연구)

  • Kim Hag-Yeol;Ahn Jae-Sung
    • Spatial Information Research
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    • v.13 no.1 s.32
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    • pp.1-17
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    • 2005
  • This paper draws new guidelines on the current method in Land Suitability Assessment (LSA), enforced to identify potential of parcels .elated to agricultural productivity, preservable value, and economic viability. In order to make recommendations of current LSA methodology, this study analyzes the issues related to the parcel-based LSA and addresses its inappropriateness f3r incorporating characteristics of each parcel. Through case studies and sensitivity analyses using GIS, the results show that (1) the $10\times10m$ grid-based LSA is much better assessment method than parcel-based one, because it gives detailed informations on attributes of each parcel; (2) more than one-distance factor should be taken into account to calculate the appropriate potential measure; and (3) the characteristics of public facilities or urbanized areas including size, density, and land use should be incorporated into the attraction variable of ny Potential model. Finally future research areas are outlined.

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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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    • v.65 no.6
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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.

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

  • Kim, Hyunjung;Jung, Chuleui;Goo, Taewon;Yi, Hoonbok
    • Korean journal of applied entomology
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    • v.53 no.3
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    • pp.199-207
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    • 2014
  • It is true that the proper environmental risk assessments for many GM (Genetically Modified) insects almost have not been executed in Korea. Therefore, we tested the environmental risk assessment about GM silkworms if there is any difference between GM silkworms and non-GM silkworms by the following three measurements. First, we measured their mobility in the breeding environment conditions with food and without food. Secondly, we measured their viability at the artificial extreme environmental conditions (low and high temperature and humidity, absent/present of foods,) after escaping from their breeding environments. Thirdly, we observed the number of laying eggs and their hatchability between GM silkworms and non-GM silkworms with four different pair experiments. The mobility of GM silkworms and non-GM silkworms statistically did not differ, and the egg productivity and hatchability were not also different. The hatchability by couple of GM female silkworms and non-GM male silkworms was lower than by non-GM male and female couple between the GM silkworms and non-GM silkworms, and there was statistically different. Relatively, the viability of GM silkworms was lower than non-GM silkworms. We could not exactly test for viability of silkworms in low temperature conditions because of their hibernating. Although there was any difference in viability and hatchability between GM silkworms and non-GM silkworms, all ability of GM silkworms was lower than non-GM silkworms. Conclusively, the environmental risk of GM silkworm was relatively lower than non-GM silkworm in this study.