• Asian Pacific Journal of Cancer Prevention
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• 제15권2호
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• pp.983-987
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• 2014

Nigella sativa (N sativa), commonly known as black seed, has been used in traditional medicine to treat many diseases. The antioxidant, anti-inflammatory, and antibacterial activities of N sativa extracts are well known. Therefore, the present study was designed to investigate the anticancer activity of seed extract (NSE) and seed oil (NSO) of N sativa against a human lung cancer cell line. Cells were exposed to 0.01 to 1 mg/ml of NSE and NSO for 24 h, then percent cell viability was assessed by 3-(4, 5-dimethylthiazol-2yl)-2, 5-biphenyl tetrazolium bromide (MTT) and neutral red uptake (NRU) assays, and cellular morphology by phase contrast inverted microscopy. The results showed NSE and NSO significantly reduce the cell viability and alter the cellular morphology of A-549 cells in a concentration dependent manner. The percent cell viability was recorded as 75%, 50%, and 26% at 0.25, 0.5, and 1 mg/ml of NSE by MTT assay and 73%, 48%, and 23% at 0.25, 0.5, and 1 mg/ml of NSE by NRU assay. Exposure to NSO concentrations of 0.1 mg/ml and above for 24 h was also found to be cytotoxic. The decrease in cell viability at 0.1, 0.25, 0.5, and 1 mg/ml of NSO was recorded to be 89%, 52%, 41%, and 13% by MTT assay and 85%, 52%, 38%, and 11% by NRU assay, respectively. A-549 cells exposed to 0.25, 0.5 and 1 mg/ml of NSE and NSO lost their typical morphology and appeared smaller in size. The data revealed that the treatment of seed extract (NSE) and seed oil (NSO) of Nigella sativa significantly reduce viability of human lung cancer cells.

• 한국식품조리과학회지
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• 제27권4호
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• pp.67-73
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• 2011

This study was performed to assess the neuroprotective effects of methanolic extracts from sweet persimmon peel (PPE) against glutamate-induced cytotoxicity in hybridoma N18-RE-105 cells. The neuroprotective effects of PPE in N18-RE-105 cells were measured using the MTT reduction assay, LDH release assay, and phase-contrast microscopy. The results of the MTT reduction assay showed that treating cells with 500 ${\mu}g/ml$ PPE resulted in cell viability of 66.9%. Additionally, the morphological changes and the results of the LDH release assay showed that glutamate-induced damage to nerve cells was strongly inhibited by PPE. GSH content of N18-RE-105 cells was 3.5 ${\mu}M$ compared to that of the control, whereas pretreatment with 500 ${\mu}g/ml$ PPE increased GSH content by 4.7 ${\mu}M$. PPE was fractionated with hexane, and that layer had the highest neuroprotective effects in glutamate-stressed N18-RE-105 cells. In conclusion, our data showed that glutamate potentiated the effects of N18-RE-105 cell death by a mechanism involving oxidative stress. Therefore, PPE may be a potential candidate for prevention and therapy of neurodegenerative diseases.

• The Korean Journal of Physiology and Pharmacology
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• 제19권4호
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• pp.357-363
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• 2015

The purpose of this study was to compare the inhibitory effect of bevacizumab on human Tenon's fibroblasts (HTFs) cultured from primary and recurrent pterygium. Cultured HTFs were exposed to 2.0, 5.0, 7.5, and 15.0 mg/mL concentration of bevacizumab for 24 hours. The 3-[4,5-dimethylthiazol- 2-yl]-2,5-diphenyl tetrazolium bromide and lactate dehydrogenase leakage assays were then performed to assess fibroblast metabolism and viability. The matrix metalloproteinase (MMP), procollagen type I C terminal propeptide (PIP), and laminin immunoassays were performed to examine extracellular matrix production. Changes in cellular morphology were examined by phase-contrast and transmission electron microscopy. Both metabolic activity and viability of primary and recurrent pterygium HTFs were inhibited by bevacizumab in a dose-dependent manner, especially at concentrations greater than 7.5 mg/mL. Both types of HTFs had significant decreases in MMP-1, PIP, and laminin levels. Distinctly, the inhibitory effect of bevacizumab on MMP-1 level related with collagenase in primary pterygium HTFs was significantly higher than that of recurrent pterygium. Significant changes in cellular density and morphology both occurred at bevacizumab concentrations greater than 7.5 mg/mL. Only primary pterygium HTFs had a reduction in cellular density at a bevacizumab concentration of 5.0 mg/mL. Bevacizumab inhibits primary and recurrent pterygium HTFs in a dose-dependent manner, especially at concentrations greater than 7.5 mg/mL. As the primary HTFs produces larger amounts of MMP-1 compared to recurrent HTFs, significant reduction in MMP-1 level in primary pterygium HTFs after exposure to bevacizumab is likely to be related to the faster cellular density changes in primary pterygium HTFs.

• 한국산업보건학회지
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• 제22권3호
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• pp.224-234
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• 2012

Objectives: To assess the hazard of Korea chrysotile and anthophylite, fibers were analyzed for their physicochemical properties by transmission electron microscope equipped with energy dispersive X-ray spectrometer (TEM-EDS). Methods: To evaluate the biopersistence of 2 domestic asbestos, Sprague-Dawely rats were exposed to 2 mg asbestos by intratracheal instillation. Each asbestos (chrysotile ; $8,814,244{\times}10^6$ fibers/mg, average size $0.08{\mu}m{\times}4.39{\mu}m$, anthophyllite ; $5,182{\times}10^6$ fibers/mg, average size $0.95{\mu}m{\times}7.29{\mu}m$) were evaluated after a single intratracheal instillation. At times from 1 week to 4 weeks after exposure, the numbers of asbestos fivers in the bronchoalveolar lavage fluid and in the lung were calculated. Results: Anthophyllite fivers continuously have retained for 4 weeks but chrysotile fivers were rarely found at 4 weeks after exposure in the bronchoalveolar lavage fluid. Chrysotile fivers at 4 weeks after treatment were not observed but anthophyllite was easily observed in the lung with phase contrast microscopy. According to electron microscopic observation of asbestos in the lung, within 1 week after the administration of chrysotile fivers were decreased rapidly but anthophyllite fivers were very little change for 4 weeks. When chrysotile fivers have been lost Fe in 1 week, there were no significant changes in anthophyllite fivers in the lung. Conclusions: These findings indicate that after a long time exposure to chrysotile, asbestos bodies can not be found in the bronchoalveolar lavage fluid.

• Toxicological Research
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• 제28권4호
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• pp.209-216
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• 2012

The biological activity of particles is largely dependent on their size in biological systems. Dispersion in the aqueous phase has been both a critical impediment to and a prerequisite for particle studies. Carbon black has been used as a surrogate to investigate the biological effects of carbonaceous particles. Here, biocompatible methods were established to disperse carbon black into ultrafine and fine particles which are generally distinguished by the small size of 100 nm. Carbon black with a distinct particle size, N330 and N990 were suspended in blood plasma, cell culture media, Krebs-Ringer's solution (KR), or physiological salt solution (PSS). Large clumps were observed in all dispersion preparations; however, sonication improved dispersion - averaged particle sizes for N330 and N990 were $85.0{\pm}42.9$ and $112.4{\pm}67.9$ nm, respectively, in plasma; the corresponding sizes in culture media were $84.8{\pm}38.4$ and $164.1{\pm}77.8$ nm. However, sonication was not enough to disperse N330 less than 100 nm in either KR or PSS. Application of Tween 80 along with sonication reduced the size of N330 to less than 100 nm, and dispersed N990 larger than 100 nm ($73.6{\pm}28.8$ and $80.1{\pm}30.0$ nm for N330 and $349.5{\pm}161.8$ and $399.8{\pm}181.1$ nm for N990 in KR and PSS, respectively). In contrast, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) exhibited little effect. Electron microscopy confirmed the typical aciniform structure of the carbon arrays; however, zeta potential measurement failed to explain the dispersibility of carbon black. The methods established in this study could disperse carbon black into ultrafine and fine particles, and may serve as a useful model for the study of particle toxicity, particularly size-related effects.

• Journal of Nutrition and Health
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• 제53권6호
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• pp.563-569
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• 2020

Purpose: The vascular smooth muscle cells (VSMCs) in mature animals have implicated to play a major role in the progression of cardiovascular diseases such as atherosclerosis. This study aimed at optimizing the protocol in culturing primary VSMCs (pVSMCs) from rat thoracic aorta and investigating the effect of cellular zinc (Zn) deficiency on cell proliferation of the isolated pVSMCs. Methods: The thoracic aorta from 7-month-old Sprague Dawley rats was isolated, minced and digested by the enzymatic process of collagenase I and elastase, and then inoculated with the culture Dulbecco Modified Eagle Medium (DMEM) at 37℃ in an incubator. The primary cell culture morphology was observed using phase-contrast microscopy and cellular Zn was depleted using Chelex-100 resin (extracellular zinc depletion only) or 3 µM N,N,N',N'-tetrakis(2-pyridinylmethyl)-1,2-ethanediamine (TPEN) (extracellular and intracellular zinc depletion). Western blot analysis was used for the detection of SM22α and calponin as smooth muscle cell marker proteins and von Willebrand factor as endothelial cell marker protein to detect the culture purity. Cell proliferation by Zn depletion (1 day) was measured by MTT assay. Results: A primary culture protocol for pVSMCs from rat thoracic aorta was developed and optimized. Isolated cultures exhibited hill and valley morphology as the major characteristics of pVSMCs and expressed the smooth muscle cell protein markers, SM22α and calponin, while the endothelial marker von Willebrand factor was hardly detected. Zn deprivation for 1 day culture decreased rat primary vascular smooth muscle cell proliferation and this pattern was more prominent under severe Zn depletion (3 µM TPEN), while less prominent under mild Zn depletion (Chelexing). Conclusion: Our results suggest that cellular Zn deprivation decreased pVSMC proliferation and this may be involved in phenotypic modulation of pVSMC in the aorta.

• 한국구조물진단유지관리공학회 논문집
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• 제27권1호
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• pp.103-108
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• 2023

이 연구에서는 자발적 균열 치유 콘크리트에 적용할 수 있는 미생물자원을 확보하기 위한 기초 연구를 수행하였다. 이를 위해 본 실험에서는 생체광물 형성 미생물을 시료에서 분리하고 시멘트 내부 생존 및 탄산칼슘 석출량을 비교하여 적합한 미생물자원을 확보하였다. 시료에서 내생포자(endospore)를 형성하는 Bacillus 계열의 박테리아를 분리하여 16S rRNA 염기서열 분석법으로 동정한 6종의 미생물이 생성하는 탄산칼슘 석출량을 비교하였다. 탄산칼슘 석출량이 가장 많은 Bacillus velezensis와 Bacillus subtilis의 2종의 미생물을 선별하였고, 모르타르에 첨가 후 양생하여 위상차 현미경 관찰을 통해 미생물의 생존을 확인하였다. 또한 모르타르에 인위적 균열을 발생시켜 미생물에 의해 생성된 균열치유물질에 의한 자발적 균열 치유 작용을 확인할 수 있었다.

• Journal of Periodontal and Implant Science
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• 제29권2호
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• pp.311-326
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• 1999

본 연구에서는 치주질환과 관련이 깊은 것으로 알려진 구강내 spirochetes 균 중 Treponema denticola(TDC)와 가장 최근에 분리 배양된 Treponema lecithinolyticum(TLC)이 치주인대세포에 미치는 영향을 알아보기 위하여 두 spirochtes 균주를 배양한후 MTT test를 이용한 치주인대 세포의 증식 억제효과를 알아보았다. 또한, 위상차 현미경을 이용한 세포형태의 변화를 관찰 하였으며, LDH(lactate dehydrogenase) test를 이용한 세포독성 실험과, gelatin zymography를 시행하여 교원질 분해효소의 하나인 gelatinase의 활성화 여부를 측정한 결과 다음과 같은 결론을 얻었다. 1. 일정한 반응시간에서 농도에 따른 세포증식 억제효과에서는 TLC의 경우 높은 농도$(150{\mu}g/well)$에서부터 세포증식 억제 효과가 나타났으며, TDC에서는 낮은 농도$(9.4{\mu}g/well)$에서도 억제 효과가 나타났다. 2. 일정한 농도에서 시간에 따른 세포증식 억제효과에서는 TLC의 경우 2일째 $150{\mu}g/well$ 농도에서부터 세포증식 억제효과가 나타났으며, TDC에서는 $9.4{\mu}g/well$ 농도에서는 2일째에 세포증식 억제효과가 나타났다. 3. 열처리한 세균의 세포증식 억제 효과에서는 TDC의 경우에서 열처리 시킨 경우와 그렇지 않은 경우에 차이가 나타났으나, TLC의 경우에는 차이가 없었다. 4. 위상차현미경으로 관찰한 치주인대세포의 형태변화는 대조군에 비해 실험군에서 세포 형태의 손상으로 방추형이 소실되었고 세포증식이 억제되었으며, 세포끼리의 연결 또한 끊어져 분리되어 있었다. 5. 세포독성을 알아보기 위한 LDH test에서는 대조군, 실험군 모두 큰 차이가 없었다. 6. Zymography를 통한 교원질 분해에 미치는 영향에서는 TDC와 TLC에 의한 분자량 72kDa의 progelatinase A가 활성형으로 발현되었다. 이상의 결과를 보아 TLC와 TDC는 세포증식 억제효과를 통해 치주인대세포에 영향을 미치며, 교원질 분해 효소 Type IV의 하나인 progelatinase A를 활성형으로 발현시킴을 확인하였다.

• 한국응용곤충학회지
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• 제47권4호
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• pp.457-465
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• 2008

본 연구는 모기에 활성을 나타내는 Bacillus thuringiensis subsp. tohokuensis CAB167균주에서 생산된 내독소결정단백질의 특성을 조사하였다. B. thuringiensis subsp. tohokuensis CAB167균주는 위상차 현미경과 주사전자현미경으로 관찰하여 일반적으로 모기에 활성을 나타내는 spherical type의 내독소결정 성단백질을 형성하는 것으로 나타났다. 국내에서 서식하는 지하집모기(Culex pipiens molestus), 빨간집모기(Culex pipiens pallens), 이집트숲모기(Aedes aegyti)에 대한 이 균주의 살충활성검정은 각각 173, 190, 580 ng/ml의 $LC_{50}$ 값으로 각각 활성을 보였다. B. thuringiensis subsp. tohokuensis CAB167균주에서 생산된 내독소결정성단백질의 SDS-PAGE를 통해 135, 80, 49와 28-kDa의 주요한 4개의 밴드를 나타냈다. 내독소결정성단백질의 소화에 영향을 미치는 곤충중장액과 유사한 trypsin효소를 처리하여 72와 63-kDa의 단백질이 새롭게 생성하였다. 파리목 위생해충인 모기에 활성을 나타내는 다른 7개의 B. thuringiensis 균주들의 내독소 결정성단백질과 혈청학적인 비교실험에서 독소단백질이 차이가 있는 것을 확인하였다.

• Journal of Periodontal and Implant Science
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• 제23권1호
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• pp.109-126
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• 1993

Refractory periodontitis manifest progressive attachment loss in a rapid and unrelenting manner regardless of the type or frequency of therapy applied. The purpose of this study was ta evaluate the relation between the level of cytokines in GCF and periodontopathic microflora with disease activity of refractory periodontitis. Selection of patients with refractory periodontitis (7 males, 3 females) were made by long term clinical observation including conventional clinical history and parameters. Teeth that showed pocket depth greater than 6mm were selected as sample teeth. Subjects were examined at baseline and after 3 months. Prior to baseline test, individual acrylic stent was fabricated. Reference grooves were made on each sample tooth site. Pocket depth and attachment loss were measured by Florida Probe. Gingival index was measured at 4 sites each sample teeth. Disease activity was defined as attachment loss of ${\ge}$ 2.1mm, as determined by sequential probing and tolerance method. The pattern and amount of alveolar bone resorption was observed with quantitative digital subtraction image processing radiography. Morphological analysis of subgingival bacteria was taken by phase contrast microscopy. Predominant cultivable bacterial distribution and frequency were compared between disease-active and disease-inactive site using immunofluorescence microscopy and selective microbial culturing. Levels of $interleukin-l{\beta}$, 2, 4, 6 and $TNF-{\alpha}$ in GCF and blood serum sample were quantified by ELISA. In active sites, P. intermedia was significantly increased to compare with inactive site. $IL-1{\beta}$, IL-2, IL-6 and $TNF-{\alpha}$ in GCF were increased in active sites and IL-2 in serum was increased in active patients significantly. Alveolar bone loss in active site was correlated with $IL-1{\beta}$, IL-2 in GCF. And loss of attachment in active site was correlated with IL-2 in GCF. These results demonstrate that IL-2 in serum, $IL-1{\beta}$, IL-2, IL-6 and $TNF-{\alpha}$ in GCF, P, intermedia might be used as possible predictors of disease activity in refractory periodontitis before it is clinically expressed as attachment loss and quantitative alveolar bone change.