• Biomolecules & Therapeutics
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• 제26권6호
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• pp.608-615
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• 2018

Benzalkonium chloride, diazolidinyl urea, and imidazolidinyl urea are commonly used preservatives in cosmetics. Recent reports suggested that these compounds may have cellular and systemic toxicity in high concentration. In addition, diazolidinyl urea and imidazolidinyl urea are known formaldehyde (FA) releasers, raising concerns for these cosmetic preservatives. In this study, we investigated the effects of benzalkonium chloride, diazolidinyl urea, and imidazolidinyl urea on ROS-dependent apoptosis of rat neural progenitor cells (NPCs) in vitro. Cells were isolated and cultured from embryonic day 14 rat cortices. Cultured cells were treated with 1-1,000 nM benzalkonium chloride, and $1-50{\mu}M$ diazolidinyl urea or imidazolidinyl urea at various time points to measure the reactive oxygen species (ROS). PI staining, MTT assay, and live-cell imaging were used for cell viability measurements. Western blot was carried out for cleaved caspase-3 and cleaved caspase-8 as apoptotic protein markers. In rat NPCs, ROS production and cleaved caspase-8 expression were increased while the cell viability was decreased in high concentrations of these substances. These results suggest that several cosmetic preservatives at high concentrations can induce neural toxicity in rat brains through ROS induction and apoptosis.

• 한국환경보건학회지
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• 제21권3호
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• pp.16-22
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• 1995

This study was designed to investigate the antibody production to sheep red blood cells(SRBC) and proliferation of mitogen-stimulated spleen cells in Balb/c mice which received cadmium chloride. The mice were divided into three independent groups which were one control and two experimental groups by the cadmium treatment or not. No specific treatment was done for the control group. One of two experimental groups, which is called 'pre-treatment group' in this paper, was subcutaneously injected with low dose of cadmium chloride(0.5 mg/kg/day) for 5 consecutive days before the primary SRBC immunization. The other called 'non-pretreatment group' was only pretreated with normal saline. Both experimental groups were intraperitoneally injected with high dose of cadmium chloride(5 mg/kg) 8 hours before the primary immunization. Mice were intraperitoneally immunized twice with 2% SRBC suspension containing $10^8$ cells. The results obtained were as follows, 1. The PFG responses to SRBC were significantly increased in two experimental groups, cadmium pretreatment and non-pretreatment compared with that of control group(p<0.05). 2. The total antibody titers to SRBC in cadmium treated groups were similar to that of control group, but titers of IgG antibody were significantly elevated(p<0.01). 3. The proliferation response of spleen lymphocytes to various mitogens was suppressed in proportion to the concentration of cadmium and the degree of cadmium accumulation in liver was increased in the cadmium treated groups. These results suggest that cadmium chloride could affect on mouse immune response, especially its cell mediated immune response could be decreased while its humoral immune response could be increased, which may not be influenced by the administration methods or pretreatment of cadmium to mouse.

• 콘크리트학회논문집
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• 제16권4호
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• pp.549-555
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• 2004

Cement of three alkalinities (equivalent alkalinities of 0.36,0.52 and 0.97) was employed in fabricating a set of classical G109 type specimens. To-date, these have been subjected to a one week wet-one week dry cyclic pending using 15 w/o NaCl solution. At the end of the dry period, potential and macro-cell current were measured to indicate whether the top reinforcing steel was in the passive or active state. Once this bar became active, the specimen was autopsied and the extent of corrosion was documented. Subsequent to visual inspection, concrete powder samples were collected from the upper region of the top rebar trace; and at a certain times concrete cores were taken from non-reinforced specimens. Using these, determinations were made of (1) critical chloride concentration for corrosion initiation ($Cl_{th}^-$), (2) effective chloride diffusion coefficient ($D_e$), and (3) pore water alkalinity ($[OH^-]$). The pore water alkalinity was strongly related to the alkali content of cement that was used in the mix. The chloride concentration, ($Cl^-$), was greater at active than at passive sites, presumably as a consequence of electro migration and accumulation of these species at active site subsequent to corrosion initiation. Accordingly, ($Cl^-$) at passive sites was considered indicative of the threshold concentration fur corrosion initiation. The $Cl_{th}^-$ was increased with increasing Time-to-corrosion ($T_i$). Consequently, the HA(High Alkalinity) specimens exhibited the highest $Cl_{th}^-$ and the NA(Normal Alkalinity) was the least. This range exceeds what has previously been reported in North America. In addition, the effective diffusion coefficient, $D_e$, was about 40 percent lower for concrete prepared with the HA cement compared to the NA and LA(Low Alkalinity) ones.

• 한국건설순환자원학회논문집
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• 제2권1호
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• pp.34-39
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• 2014

이 연구의 목적은 하이볼륨 플라이애시(HVFA) 콘크리트의 강도수준 및 플라이애시의 혼입률별로 콘크리트 내의 철근부식 및 염소이온 침투 저항성을 평가하는 것이다. 이를 위하여 물-결합재비 및 플라이애시 혼입률에 따라 철근 상부를 노출시킨 원주형 공시체와 통상적인 원주형 공시체를 제작하였으며, 이들 각각에 대해 압축강도 및 염소이온 침투 저항성 시험과 전기화학적 방법에 의한 철근부식 촉진시험을 수행하였다. 실험 결과, 대체적으로 플라이애시 혼입률이 많을수록 HVFA 콘크리트의 압축강도는 감소하였으나, 철근부식 및 염소이온 침투 저항성은 크게 개선되는 것으로 나타났다.

• 공업화학
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• 제9권6호
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• pp.811-816
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• 1998

염분을 함유한 시멘트 모르터 내의 철근의 부식현상을 교류 임피던스법에 의해 고찰하였다. 부식 가속화 장치를 이용하여 단기간 내에 부식현상을 예측하였으며, 측정된 임피던스 값은 제안된 전기화학적 등가회로 및 모델에 적용할 수 있었으며, CNLS(complex nonlinear least squares) fitting법에 의하여 계산된 값과 실험에서 얻은 값이 잘 일치함을 알 수 있었다. 주어진 모델로부터 구한 전하이동저항 (charge transfer resistance, $R_2$)은 염분 농도와 시간에 따른 철근의 무게 감소량의 예측을 가능하게 하였으며, 이는 실제 철근의 무게 감소치에 근접함을 알 수 있었다.

• 한국식품영양학회지
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• 제13권5호
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• pp.460-464
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• 2000

본 연구에서는 전통적으로 담석증, 신장치료 등의 한약제로 많이 사용하는 대왕을 여러 용매를 사용하여 얻은 대왕추출물 분액에 대한 세포독성을 여부를 MTT 정량법, NR 정량법, SRB 정량법을 이용하여 조사하였다. 1. 추출 용매 methylene chloride, ethyl acetate, butanol, water로부터 얻은 대왕추출물 모두 처리농도에 따라 세포에 미치는 영향이 증가하였다. 2. Butanol을 용매로 사용하여 얻은 대왕추출물 분액이 다른 3가지 용매로부터 얻은 대왕추출물보다 세포에 미치는 영향이 크게 나타났고 water를 용매로 사용하여 얻은 추출물이 A498 세포주에 미치는 영향이 가장 낮게 나타났다. 3. Butanol을 추출 용매로 하여 얻은 대왕추출물이 A498 세포주에 미치는 영향이 가장 컸는데 그 추출물에 대한 MTT$_{50}$, NR$_{50}$, SRB$_{50}$값은 각각 0.63mg/ml, 0.65mg/ml, 0.68mg/ml이었고, 가장 영향이 적은 water의 경우 MTT$_{50}$, NR$_{50}$, SRB$_{50}$값은 각각 0.84mg/ml, 0.82mg/ml, 0.80mg/ml이었다. 4. 정량방법 간의 대왕추출물에 대한 반응은 MTT 정량법이 가장 민감하게 나타났다.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.292-292
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• 2017

This study was conducted to evaluate the effect of anti-obesity and antioxidant enzyme activities in vitro by different solvent fractions from the roots of Codonopsis lanceolata. The cytotoxicity of different solvent fractions of C. lanceolata on 3T3-L1 preadipocytes were evaluated using the MTT assay, the rate of cell survival progressively decreased in a dose-dependent manner. Butyl alcohol fraction at $200{\mu}g/mL$ exhibited a pronounced cytotoxic effect (75.73%) on 3T3-L1 cell comparable to that of the hexane fraction (79.82%), methylene chloride fraction (84.02%), ethyl acetate fraction (87.62%) and DW fraction (86.30%) at the same concentration. The Oil Red O solution was used to determine whether different solvent fractions of C. lanceolata induce adipocyte differentiation in 3T3-L1 preadipocytes. Confluent 3T3-L1 cells were treated with $50{\mu}g/mL$ concentration of solvent fraction extracts from C. lanceolata. Inhibitory degree of lipid accumulation against solvent fraction extracts showed a significant level compared with the control. Both lipid accumulation and adipocyte differentiation showed relatively high effect on methyl chloride fraction. The root extract of C. lanceolata had the highest SOD enzyme activity of 84.5% in ethyl acetate partition layer and while water partition layer of diploid showed the lowest SOD enzyme activity of 57.9%. The activity of CAT, APX and POD showed a significantly higher activity in ethyl acetate partition layer compared with the other fraction. These results suggested that the roots of C. lanceolata may assist in the potential biological activity on anti-obesity and antioxidant capacity.

• Biomolecules & Therapeutics
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• 제15권4호
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• pp.261-265
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• 2007

Hydroxyproline (HYP) is a post-translational product of proline hydroxylation catalyzed by an enzyme prolyl 4-hydroxylase (P4H) which plays a crucial role in the synthesis of all collagens. Considering the role of collagen and its significance in many clinically important diseases such as liver fibrosis, a great deal of attention has been directed toward the development of an assay at cell-based system. The reason is that cell-based assay system is more efficient than enzyme-based in vitro system and takes much less time than in vivo system. Several assay procedures developed for P4H are laborious, time-consuming and not feasible for the massive-screening. Here, we report the cell-based assay method of prolyl 4-hydroxylase in immortalized rat hepatic stellate HSC-T6 cells. To optimize the cell culture condition to assay for HYP content, various concentrations of reagents were treated for different times in HSC-T6 cells. Our data showed that the treatment with ascorbate in a hypoxic condition for 24 h resulted in the maximal increase of HYP by 1.8 fold. Alternatively, cobalt chloride ($5\;{\mu}M$) and ascorbate ($50\;{\mu}M$) in normoxic states exhibited similar effect on the production of HYP as in a hypoxic condition. Therefore, cobalt chloride can be substituted for a hypoxic condition when an anaerobic chamber is not available. Rosiglitazone and HOE077, known as inhibitors of collagen, synthesis decreased P4H enzyme activity by 32.3% and 15%, respectively, which coincided with previous reports from liver tissues. The level of the smooth muscle ${\alpha}$-actin, a marker of activated stellate cells, was significantly increased under hypoxia, suggesting that our experimental condition could work for screening the anti-fibrotic compounds. The assay procedure took only 3 days after treatment with agents, while assays from the primary stellate cells or liver tissues have taken several weeks. Considering the time and expenses, this assay method could be useful to screen the compounds for the inhibitor of prolyl 4-hydroxylase.

• 한방안이비인후피부과학회지
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• 제23권2호
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• pp.1-12
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• 2010

Objective : For this study, mice on mercurial toxication were given mercuric subcutaneous injection to their abdomen factitiously. After delivering Kami- bangpungtongseong-san(KBT) extracts to the mice by oral administration, we observed changes from liver and kidney of mice. Method : The BALB/c mice were distributed into three groups: No treated group(Normal group), Mercuric chloride subcutaneous injection group(Control group), Kami-bangpungtongseong-san-treated group (Sample group). KBT Extracts were delivered orally in 7 days. We observed involution of liver, necrosis of liver and cell plate loss of liver, lipid peroxidation CYP1A1 expression. We observed involution of proximal convoluted tubules, hypertrophy of Bowman's capsule, periodic acid-Schiff(PAS)'s positive reaction of proximal convoluted tubules, heat shock protein(HSP)700's positive reaction in glomerulus. For the charting the results, image analysis was taken. The result of image analysis was verified significance by Sigmaplot 2000(P<0.05). Result : The mice' liver on mercurial toxication were relieved involution of liver, necrosis of liver, and cell plate loss of liver and also declined lipid peroxidation and CYP1A1 expression. The mice' kidney on mercurial toxication were relieved involution of proximal convoluted tubules, hypertrophy of Bowman's capsule and increasing PAS's positive reaction of proximal convoluted tubules. On the other hand it was declined HSP700's positive reaction in glomerulus. Conclusion : According to the result of study, we think that we can expect to the effect of KBT extracts' therapeutic action to tissue injuries of the mice' liver and kidney on acute mercurial toxication.

• Asian Pacific Journal of Cancer Prevention
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• 제16권13호
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• pp.5371-5376
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• 2015

Berberine (B1), isolated from stems of Coscinium fenestratum (Goetgh.) Colebr, was used as a principle structure to synthesize three phenolic derivatives: berberrubine (B2) with a single phenolic group, berberrubine chloride (B3) as a chloride counter ion derivative, and 2,3,9,10-tetra-hydroxyberberine chloride (B4) with four phenolic groups, to investigate their direct and indirect antioxidant activities. For DPPH assay, compounds B4, B3, and B2 showed good direct antioxidant activity ($IC_{50}$ values=$10.7{\pm}1.76$, $55.2{\pm}2.24$, and $87.4{\pm}6.65{\mu}M$, respectively) whereas the $IC_{50}$ value of berberine was higher than $500{\mu}M$. Moreover, compound B4 exhibited a better DPPH scavenging activity than BHT as a standard antioxidant ($IC_{50}=72.7{\pm}7.22{\mu}M$) due to the ortho position of hydroxyl groups and its capacity to undergo intramolecular hydrogen bonding. For cytotoxicity assay against human fibrosarcoma cells (HT1080) using MTT reagent, the sequence of $IC_{50}$ value at 7-day treatment stated that B1 < B4 < B2 ($0.44{\pm}0.03$, $2.88{\pm}0.23$, and $6.05{\pm}0.64{\mu}M$, respectively). Berberine derivatives, B2 and B4, showed approximately the same level of CAT expression and significant up-regulation of SOD expression in a dose-dependent manner compared to berberine treatment for 7-day exposure using reverse transcription-polymerase chain reaction (RT-PCR) assays. Our findings show a better direct-antioxidant activity of the derivatives containing phenolic groups than berberine in a cell-free system. For cell-based system, berberine was able to exert better cytotoxic activity than its derivatives. Berberine derivatives containing a single and four phenolic groups showed improved up-regulation of SOD gene expression. Cytotoxic action might not be the main effect of berberine derivatives. Other pharmacological targets of these derivatives should be further investigated to confirm the medical benefit of phenolic groups introduced into the berberine molecule.