• Title/Summary/Keyword: Incubation Effect

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The Optimization of Method for Prediction of Drug-Induced Liver Injury Using HepG2 Cells Cultured with Human Liver Microsomes (Human Liver Microsomes과 HepG2 세포를 이용한 약물유래 간독성 평가 방법의 최적화)

  • Choi, Jong Min;Jeon, Jang Su;Kim, Sang Kyum
    • YAKHAK HOEJI
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    • v.59 no.5
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    • pp.201-206
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    • 2015
  • The aim of the present study was to optimize in vitro method for the prediction of drug-induced liver injury using human liver microsomes (HLM). Cytotoxicity test of cyclophosphamide and acetaminophen in HepG2 cells cultured with HLM showed that the newly established condition using 0.375 mg/ml HLM for 24 hr incubation was comparable or more sensitive than the previously established condition using 0.75 mg/ml HLM for 12 hr incubation. Although the cytotoxic effect of troglitazone was completely attenuated by 0.75 mg/ml HLM, it was augmented by 0.375 mg/ml HLM in the presence of the NADPH-generating system. The cytotoxic effect of chlormezanone, a withdrawn drug due to hepatotoxicity in human, was increased by HLM in the presence of the NADPH-generating system. In contrast, the cytotoxic effect of methapyrilene, a withdrawn drug due to hepatotoxicity in rats, was decreased by HLM in the presence of the NADPH-generating system. The present study suggests that the optimized in vitro method using HLM can be useful for the prediction of drug-induced hepatotoxicity.

Effect of $Ca^{2+}$ on Phvsiological Activites of the Acrosome Reaction on Spermatozoa ($Ca^{2+}$이 정자 첨체반응의 생리적 활성에 미치는 영향)

  • 장재호;오영근
    • The Korean Journal of Zoology
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    • v.39 no.2
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    • pp.182-189
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    • 1996
  • It has been known that spermatozoa should obtain their fertilizing ability through capacitation and acrosome reaction, and that in these processes of fertilization, Ca2+ platys an important role for their conjugation. Therefore the present study has been designed in order to clarify the effect of fluctuation of the media Ca2+ level and the intracellular concentration of the spermatozoa on the acrosomes. During the incubation of spermatozoa, a considerable fluctuation in the media Ca2+ level has been observed after the BSA administration and the media concentration of Ca2+. It is deduced that these fluctuation rates may have an effect on the acrosome reaction. The fluctuation of K+ flux has been observed in accordance with the incubation period over time, and it's concentration seems to be closely related with the acrosomal reaction. The respiratory exchange rate (RERI of the spermatozoa is kept more regular in the BSA and Cacl2 administration groups than the non-administration group. Based on the experimental findings, it is possible to deduce a hypothesis from these findings that physiological activities of the acrosome reaction are not functionally related to the media Ca2+ level and the intracellular influx of Ca2+ concentration, although Ca2+ platys an important role as a stimulating factor in the acrosome reaction.

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The Effect of Extract of Cervi pantotrichum Cornu on Human Sperm Motility (녹용(Cervi pantotrichum cornu) 추출물이 인간 정자 운동성에 미치는 영향)

  • 신동혁;김지영;이창훈;정창진;조용선;최동희;고정재;윤태기
    • Journal of Embryo Transfer
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    • v.17 no.3
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    • pp.187-193
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    • 2002
  • This study was conducted to determine the effect of extract of Cervi pantotrichum cornu on human sperm motility, Four different types of media were prepared such as plain Ham's F-10 medium(control medium), control medium containing 0.3% bovine serum albumin(BSA)(medium A), control medium containing the extract of Cervi pantotrichum cornu aqua-acupuncture medium(medium B) and medium B containing 0.3% BSA(medium C). Human semen were washed and divided into 4 fractions and sperm were cultured in those medium for up to 72 hours at 37$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$ in air. A total twenty eight semen samples including 14 normozoospermia and 14 asthenospermia were used for this study. In normozoospermia group, motility of control medium and medium A, B and C were 4.1%, 1.3%, 64.5%, and 77.1%, respectively after 24 hours of incubation, and were 0.0%, 0.0%. 8.8% and 44.9%, respectively after 48 hours of incubation. In asthenospermia group, motility of control medium and medium A, B and C were 2.0%, 2.2%, 58.3% and 85.1%, respectively after 24 hours of incubation, and decreased to 0.0%, 0.2%, 5.8% and 29.6%, respectively after 48 hours of incubation. In both groups, highest sperm motility was observed in medium C group when compared with other media. Furthermore motile sperm were found in medium C after 72 hours of incubation while no motile sperm was observed in the other media. Therefore it could be concluded that the extract of Cervi pantotrichum rornu affects on the human sperm motility.

The Effect of Nitroprusside on the Sperm Motility, Viability, and Reactive Oxygen Species Generation (Nitroprusside가 인간정자의 생존력, 운동성, Reactive Oxygen Species 발생에 미치는 영향)

  • Min, Bu-Kie;Lee, Hee-Min;Kim, Ki-Seok;Lee, Hee-Sup;Kim, Heung-Gon;Hong, Gi-Youn;Lee, Bong-Ju
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.3
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    • pp.351-355
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    • 1996
  • Objective: To analyze the direct effect of nitre oxide, generated from sodium nitroprusside, on sperm motility and reactive oxygen species. Design: Human sperm samples were treated to allow swim-up and washing. And the samples were devided into four aliquots. Each aliquot was incubated with either concentration at 0, 100nM, $10{\mu}M$, 1mM of nitroprusside. Intervention: Samples were measured chemiluminosence for reactive oxygen species of each aliquot with concentrations at 0, 100nM, $10{\mu}M$, 1mM of nitroprusside at allowing swim-up and washing of sperm. Main Outcome Measures: Percent motion parameters and viability were asse-ssed at 0, 3, 6, 12, 24 hours incubation. Results: The percent viablity was lower slightly in control group (50.2%) than that in sperm treated with 100nM of nitroprusside(57.5%) at 24 hours after incubation, while was reduced significantly in sperm with concentra-tion of $10{\mu}M(42.1%)$ and 1mM(21.3%)of nitroprusside at 6 hours after incubation. And the sperm treated with 1mM of nitroprusside was immotile totally at 6 hours after incubation. The straight line$(35.3{\pm}5.6%)$, the rapid forward$(37.2{\pm}6.4%)$ and the weak curvilinear velocity$(9.6{\pm}2.4%)$were more favorable comparing with those ($32.4{\pm}4.2%$, $30.0{\pm}7.8%$ and $18.0{\pm}4.6%$ respectively) in control group at 3 hours after incubation, but reduced significantly in sperm treated with $10{\mu}M$ and 1mM of nitroprusside. The levels of reactive oxygen species in control(700 c.p.m.) is lower significantly than that in each experimental groups of sperm treated with nitroprusside. And the levels of reactive oxygen species were 2200 c.p.m. in 100nM, 6200c.p.m. in $1{\mu}M$ and 12800c.p.m. in 1mM respectively. Conclusion: These results suggested that the concentration of 100nM of nitroprusside on sperm is beneficial to the maintanance of viablity and motile velocity, but detriment in high concentration of $10{\mu}M$ or 1mM of nitroprusside.

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Toxicity Decrease of Cadmium by the Pigment Produced by Azomonas agilis PY101 in the Culture of Bacterial Cells and Vero Cells

  • You, Kyung-Man;Lee, Soo-Youn;Park, Yong-Keun
    • Korean Journal of Environmental Biology
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    • v.20 no.3
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    • pp.232-236
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    • 2002
  • The morphological patterns and the cytopathogenicity time of the Vero cells induced by free $Cd^{2+}$ and pigment-bound $Cd^{2+}$ were observed by inverted microscope in order to investigate the difference of cadmium toxicity. The Vero cells induced by Hee $Cd^{2+}$ of 0.1 mM were shown to have a fatal toxic effect and the cytopathogenicity could be seen early after 6$\pm$2 hours of incubation. Partially affected cells induced by pigment-bound $Cd^{2+}$ of 0.1 mM were shown and the cytopathogenicity could be seen after 20 hours of incubation. The Vero cells grown with free 0.001 mM $Cd^{2+}$ were also affected and the cytopathogenicity could be seen after 17 hours of incubation, whereas the Vero cells grown with 0.001 mM pigment- bound $Cd^{2+}$ were unaffected. The sensitivity of Escherichia coli DH5$\alpha$ bacterial cells was also examined after a short treatment with free $Cd^{2+}$ or pigment-bound $Cd^{2+}$. About 5% of cells survived after 0.01 mM of free $Cd^{2+}$ treatment, while about 68% of cells survived after 0.01 mM of pigment- bound $Cd^{2+}$.

The Effect of Geometric Shape of Amorphous Silicon on the MILC Growth Rate (MILC 성장 속도에 비정질 실리콘의 기하학적 형상이 미치는 영향)

  • Kim Young-Su;Kim Min-Sun;Joo Seung-Ki
    • Korean Journal of Materials Research
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    • v.14 no.7
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    • pp.477-481
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    • 2004
  • High quality polycrystalline silicon is very critical part of the high quality thin film transistor(TFT) for display devices. Metal induced lateral crystallization(MILC) is one of the most successful technologies to crystallize the amorphous silicon at low temperature(below $550^{\circ}C$) and uses conventional and large glass substrate. In this study, we observed that the MILC behavior changed with abrupt variation of the amorphous silicon active pattern width. We explained these phenomena with the novel MILC mechanism model. The 10 nm thick Ni layers were deposited on the glass substrate having various amorphous silicon patterns. Then, we annealed the sample at $550^{\circ}C$ with rapid thermal annealing(RTA) apparatus and measured the crystallized length by optical microscope. When MILC progress from narrow-width-area(the width was $w_2$) to wide-width-area(the width was $w_1$), the MILC rate decreased dramatically and was not changed for several hours(incubation time). Also the incubation time increased as the ratio, $w_1/w_2$, get larger. We can explain these phenomena with the tensile stress that was caused by volume shrinkage due to the phase transformation from amorphous silicon to crystalline silicon.

Inhibitory Effect of Scutellaria barbata Don Water-extracts on Growth and DNA Incorporation of Human Cancer Cells

  • Kim, Dong-Il
    • The Journal of Korean Medicine
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    • v.27 no.4
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    • pp.162-173
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    • 2006
  • The water-extracts of Scutellaria barbata Don (SBDE) were isolated from Chinese medicinal plant sources. The extracts showed strong growth-inhibitory activity and cancer chemopreventive activity on the growth and DNA incorporation of MG63 human osteosarcoma and K562 human leukemia cell lines. The growth of human cancer cells was inhibited in the presence of the extracts (20, 50 and 100 ${\mu}$g/ml), and the effects were concentration-dependent and incubation time-dependent up to 8 days. When 50 ${\mu}$g/ml of the extracts was added to the media of MG63 and K562, cell growth after 8 days or 6 days of incubation was retarded by 93.2 to 97.3% of the control group. Morphological changes of MG63 and K562 cell lines were observed. As the concentration of the extracts increased up to 50 ${\mu}$g/ml, degree of cell aggregation decreased. Moreover, the DNA incorporation of the cells which were labeled with [3H] thymidine was significantly reduced after 3 days of incubation at $37^{\circ}C$ with the extract. Therefore, it is suggested that the extract is highly effective on inhibition of cancer cell growth. The extract also inhibited gene expression of IGF-II in transcriptional level. Since IGF-II works as a mitogenic effector on MG63 and K562 cell lines, these results suggest that the growth inhibition is in part mediated through the inhibition of IGF-II gene expression.

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Toxicity of Streptozotocin in Isolated Rat Hepatocytes (흰쥐의 분리 간세포에서 스트렙토조토신의 독성)

  • Park, Ki-Suk;Chong, Jong-Chol;Moon, Chang-Kiu;Chung, Jin-Ho
    • YAKHAK HOEJI
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    • v.36 no.1
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    • pp.80-86
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    • 1992
  • Streptozotocin (STZ) is a naturally occurring nitrosoamide used extensively to produce diabetes in experimental animals. Our previous study has demonstrated that i.v. administraton of streptozotocin induces significant red blood cell hemolmysis in rats. Since it has been reported that the highest concentration of STZ is found in the liver, the effect of STZ in freshly isolated rat hepatocytes has been investigated. STZ treatment (10 mM) did not cause significant loss of viability throughout 4 hour incubation, while high dose of STZ (300 mM) to hepatocytes resulted in complete cell death within 3 hours. Addition of 40 mM glucose to incubation medium did not potentiate STZ-induced hepatotoxicity, suggesting that STZ-induced hyperglycemia in vivo did not affect its hepatotoxicity. To investigate the mechanixm of the toxicity, intracellular total glutathione level was determined. Tratment with 10 mM STZ which was not toxic to hepatocytes led to complete depletion of intracellular glutathione level within 1 hour incubation. These results suggest that STZ-induced hepatotoxicity may be independent on the intracellular glutathione depletion.

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Effects of Clearance on the Formation of Adiabatic Shear Band in Stepped Specimen (계단시편의 간극이 단열전단밴드의 형성에 미치는 영향)

  • Yoo, Y.H.;Jeon, G.Y.;Chung, D.T.
    • Transactions of the Korean Society of Mechanical Engineers
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    • v.17 no.7 s.94
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    • pp.1700-1709
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    • 1993
  • The stepped specimen which is subjected to step loading is modeled to study the initiation and growth of adiabatic shear band using explicit time integration finite element method. Three different clearance sizes are tested. The material model for the stepped specimen includes effects of strain hardening, strain rate hardening and thermal softening. It is found that the material inside the fully grown adiabatic shear band experiences three phase of deformation, (1) homogeneous deformation phase, (2) initiation/incubation phase, and (3) fast growth phase. The second phase of deformation is initiated after sudden shear stress drop which occurs at the same time regardless of the clearance size. The incubation time prior to fast growth phase increases, as the clearance size of the stepped specimen increases. Whereas, after incubation period, the growth rate of the adiabatic shear band decreases, as the clearance size decreases. It is also found that two adiabatic shear band may develop instead of one for the smaller clearance size.

Low-Temperature Selective Epitaxial Growth of SiGe using a Cyclic Process of Deposition-and-Etching (증착과 식각의 연속 공정을 이용한 저온 선택적 실리콘-게르마늄 에피 성장)

  • 김상훈;이승윤;박찬우;심규환;강진영
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.16 no.8
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    • pp.657-662
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    • 2003
  • This paper presents a new fabrication method of selective SiGe epitaxial growth at 650 $^{\circ}C$ on (100) silicon wafer with oxide patterns by reduced pressure chemical vapor deposition. The new method is characterized by a cyclic process, which is composed of two parts: initially, selective SiGe epitaxy layer is grown on exposed bare silicon during a short incubation time by SiH$_4$/GeH$_4$/HCl/H$_2$system and followed etching step is achieved to remove the SiGe nuclei on oxide by HCl/H$_2$system without source gas flow. As a result, we noted that the addition of HCl serves not only to reduce the growth rate on bare Si, but also to suppress the nucleation on SiO$_2$. In addition, we confirmed that the incubation period is regenerated after etching step, so it is possible to grow thick SiGe epitaxial layer sustaining the selectivity. The effect of the addition of HCl and dopants incorporation was investigated.