• Title/Summary/Keyword: incubation condition

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Hypoxia Induces Paclitaxel-Resistance through ROS Production

  • Oh, Jin-Mi;Ryu, Yun-Kyoung;Lim, Jong-Seok;Moon, Eun-Yi
    • Biomolecules & Therapeutics
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    • v.18 no.2
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    • pp.145-151
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    • 2010
  • Oxygen supply into inside solid tumor is often diminished, which is called hypoxia. Many gene transcriptions were activated by hypoxia-inducible factor (HIF)-$1{\alpha}$. Here, we investigated the effect of hypoxia on paclitaxel-resistance induction in HeLa cervical tumor cells. When HeLa cells were incubated under hypoxia condition, HIF-$1{\alpha}$ level was increased. In contrast, paclitaxel-mediated tumor cell death was reduced by the incubation under hypoxia condition. Paclitaxel-mediated tumor cell death was also inhibited by treatment with DMOG, chemical HIF-$1{\alpha}$ stabilizer, in a dose-dependent manner. A significant increase in intracellular ROS level was detected by the incubation under hypoxia condition. A basal level of cell density was increased in response to 10 nM $H_2O_2$. HIF-$1{\alpha}$ level was increased by treatment with various concentration of $H_2O_2$. The increased level of HIF-$1{\alpha}$ by hypoxia was reduced by the treatment with N-acetylcysteine (NAC), a well-known ROS scavenger. Paclitaxel-mediated tumor cell death was increased by treatment with NAC. Taken together, these findings demonstrate that hypoxia could play a role in paclitaxel-resistance induction through ROS-mediated HIF-$1{\alpha}$ stabilization. These results suggest that hypoxia-induced ROS could, in part, control tumor cell death through an increase in HIF-$1{\alpha}$ level.

Influence of Dry Roasting of Whole Faba Beans (Vicia faba) and Whole Lupin Seeds (Lupinus albus) on Rumen Disappearance and Estimated Intestinal Digestion of CP Using the Optimal Three-Step In Vitro Technique in Dairy Cows

  • Yn, P.;Egan, A.R.;Lenry, B.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.7
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    • pp.1054-1062
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    • 1999
  • The effects of dry roasting whole faba beans (WFB) and whole lupin seeds (WLS) at 110, 130 or $150{^{\circ}C}$ for 15, 30 or 45 min on rumen (RDCP%), estimated intestinal (IDCP%) and total tract disappearance of CP (TDCP%) and intestinal availability (IARUCP%) of rumen undegraded CP (RUCP%) were determined. The RDCP values were estimated by in sacco technique by incubating nylon bags for 8, 12 and 24 h in the rumen of dairy cows. The IDCP and IARUCP values were estimated using a sequence of ruminal incubation, in vitro incubation in acid-pepsin for 1 h and then in pancreatin for 24 h of three-step in vitro procedure technique. Dry roasting at 130 and $150^{\circ}C$ decreased RDCP with correspondingly increasing IDCP. The IDCP value generally increased from 12.3(raw) to 8.6, 14.8 and 39.6% (WFB) and from 28.3 (raw) to 33.7, 36.2 and 56.2% (WLS) at 8 h rumen incubation; from 2.9 (raw) to 2.9, 4.6 and 23.3% (WFB) and from 19.6 (raw) to 19.0, 24.0 and 46.6% (WLS) at 12 h rumen incubation; from 1.3 (raw) to 1.9, 1.7 and 11.0% (WFB) and from 4.4 (raw) to 4.2, 10.7 and 36.7% (WLS) at 12 h rumen incubation as the temperatures rose to 110, 130 and $150{^{\circ}C}$ respectively. The TDCP values were always high and increased by time in the rumen, the average values of which were 97.9, 96.6; 99.2, 96.9 and 99.6, 98.7% for WFB and WLS, respectively, at 8, 12 and 24 h rumen incubation. But within the same retention time, TDCP was generally unchanged. The average IARUCP increased from 87.3 (raw) to 87.4, 88.7 and 92.0% (WFB); from 87.6 (raw) to 88.9, 91.5 and 93.0% (WLS) at roasting temperatures of 110, 130 and $150{^{\circ}C}$, respectively. It was concluded that dry roasting can shift the digestion of CP from rumen to the lower gastrointestinal tract without depressing the digestion of RUCP. The best processing condition in this study was dry roasting at $150{^{\circ}C}$ for 45 min in terms of effects on the disappearances and availability of CP. Research data on intestinal availability of individual amino acids need to be further investigated.

Characteristics on the Incubation of Sulfur Compound-Oxidizing Strains Separated for the Removal of Malodor (악취제거를 위하여 분리한 황화합물 산화균주의 배양특성)

  • Lim, Dong Joon;Lim, Kwang-Hee
    • Korean Chemical Engineering Research
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    • v.47 no.6
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    • pp.788-794
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    • 2009
  • Both strains of KD-1212 and DAH-1056 were isolated and identified from animal manure-contaminated soil by screening bacterial strains for the removal of sulfur compound-malodor with such substrate as sodium thiosulfate or free sulfur. Then the characteristics on the incubation of these microbes were observed under various incubating-condition such as pH, temperature, aerobic or anaerobic, substrate(sulfur compound) concentration, nitrogen and carbon source and rotating speed for mixing, and the optimum incubating condition was established. The optimum pHs of KD-1212 and DAH-1056 were 7.0 and 4.0, respectively, and their optimum temperatures were in the range of $30{\sim}35^{\circ}C$. Another autotrophic strain, ED-1138, was isolated from contaminated soil. The strain DAH-1056 excelled a strain Thiobacillus sp. IW in eliminating hydrogen sulfide during the process of malodor-biofiltration with a fixed strain. The characteristics on the incubation of strain KD-1212 were observed under various substrate-concentrations, nitrogen and carbon sources. KD-1212 favored glucose and maltose, and yeast extract as carbon sources and nitrogen source, respectively. The optimum concentrations of substrate and nitrogen source were 25 mM of sodium thiosulfate and 0.5% yeast extract, respectively for the growth of strain KD-1212.

Optimization of Skim Milk Fermentation Conditions by Response Surface Methodology to Improve ACE Inhibitory Activity Using Lactiplantibacillus plantarum K79 (반응표면법에 의한 Lactiplantibacillus plantarumK79를 이용한 ACE(Angiotensin Converting Enzyme) 억제활성 향상을 위한 탈지유 발효조건 최적화)

  • Park, Yu-Kyoung;Hong, Sang-Pil;Lim, Sang-Dong
    • Journal of Dairy Science and Biotechnology
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    • v.40 no.3
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    • pp.93-102
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    • 2022
  • This study was conducted using response surface methodology (RSM) to elucidate fermentation conditions that will optimize ACE inhibitory activity using Lactiplantibacillus plantarum K79. Four independent variables [skim milk (with 1% added glucose) concentration (6%-14%), incubation temperature (32℃-42℃), incubation time (8-24 h), and amount of added starter (0.02%-0.2%)] were evaluated using five-level central composite design and response surface methodology to determine the optimum fermentation condition. The dependent variables were angiotensin converting enzyme (ACE) inhibitory activity (the value obtained from 102 diluted supernatant), and pH. The respective coefficients of determinations (R2) were 0.791 and 0.905 for ACE inhibitory activity and pH. The maximum ACE inhibitory activity was 90% under the following conditions: 10% skim milk (with 1% added glucose) concentration, 37℃ incubation temperature, 17.8 h incubation time, and 0.2% added starter. Based on the RSM, using predicted best ACE conditions for fermentation of 13.49% skim milk (with 1% added glucose) with 0.0578% starter at 33.4℃ for 21.5 h, the predicted ACE inhibitory activity and pH values were 86.69% and 4.6, respectively. Actual ACE inhibitory activity and pH values were 85.5% and 4.58, respectively

혐기성 슬러지를 첨가한 오염 토양에서 저자 수용체 조건에 따른 디젤 분해 및 미생물 군집 변화

  • 이태호;최선열;박태주
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.09a
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    • pp.207-210
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    • 2004
  • Effect of electron accepters on anaerobic degradation of petroleum hydrocarbons by an anaerobic sludge taken from a sludge digestion tank in a soil artificially contaminated with 10,000 mg/kg soil of diesel fuel was tested. Treatments of soil with 30 mL of the digestion sludge (2,000 mg/L of vss (volatile suspended solids)) were incubated under several anaerobic conditions including nitrate reducing, sulfate reducing, methanogenic, and mixed electron accepters conditions for 120 days. Treatments with the digested sludge showed significant degradation of diesel fuel under all anaerobic conditions compare to control treatments with an autoclaved sludge and without the sludge. The amount of TPH degradation after 120days incubation was the largest in the treatment with the sludge and mixed electron accepters (75% removal of TPH) followed in order by sulfate reducing, nitrate reducing, methanegenic condition as 67%, 53%, 43%, respectively. However, the rate of TPH degradation in the nitrate- and sulfate reducing condition within 105 days were comparable with that of the mixed electron accepters condition. Microorganisms in each electron acceptor condition were plated on solid mediums containing nitrate or sulfate as sole electron acceptor and several nitrate- and sulfate reducing bacteria showed effective degradation of diesel fuel within 30 days incubations. These results suggest that anaerobic degradation of diesel fuel in soil with digested sludge is effective for practical remediation of soil contaminated with petroleum hydrocarbons.

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Phytomitogen Induced Blast Transformation of Guinea Pig: I. Effect of Concentration of Serum and Time of Incubation on the Uptake of 3H -Thymidine (Phytomitogen에 의한 기니픽 임파구의 Blast Transformation: II. 유사분열에 미치는 혈청농도와 배양시간의 효과)

  • Kim, Jong-shu
    • Korean Journal of Veterinary Research
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    • v.27 no.1
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    • pp.41-45
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    • 1987
  • The present study was carried out to investigate the optimal conditions on blastogenesis of guinea pig lymphocytes. A microculture system in conjuction with a semiautomatic multiple sample harvester was used to study the in vitro optimal condition of guinea pig lymphocytes. Careful analysis of lymphocyte transformation to concanavalin A (Con A) and lipopolysaccharide (LPS) mitogen determined optimal conditions as: (a) 10% fetal bovine serum in RPMI-1640 medium (b) 48-hour culture period.

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Plant Regeneration from Mesophyll Protoplast of Nicotiana africana (Nicotiana africana의 엽육 원형질체로부터 식물체 재분화)

  • 최상주;이승철
    • Journal of the Korean Society of Tobacco Science
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    • v.15 no.2
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    • pp.130-136
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    • 1993
  • Protoplasts of palisade cells were aseptically isolated from leaves of Nicotiono apicona Merxm. by the one step enzymatic method. Efficiency of colony formation were depended on cell density and light condition during incubation, but an intensity of 47 ft-c during a period of 2 weeks after isolation of the protoplasts in the Nagata and Takebe's medium promoted the planting efficiency. Protoplast - derived calli of N. africana can be differentiated into shoot when cultured on Murashige and skoog's medium containing IAA(0.Smg/$\ell$) and zeatin(5.0mg/$\ell$).

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In vitro 계에서의 Streptozotocin 분해 및 세포내 흡수

  • 박기숙;호지숙;정진호
    • Toxicological Research
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    • v.7 no.2
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    • pp.183-189
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    • 1991
  • Since streptozotocin has been known to be chemically stable only under acidic condition(pH 4), the spontaneous degradation and cellular uptake of streprozotocin in neutral incubation medium was determined by chemical assay. Streptozotocin levels in both phosphate buffered saline (pH 7.4) and Krebs-Henseleit buffer (pH 7.4) decreased with a half life of about 2 hours. The presence of erythrocytes or pepatocytes under the same buffers did not affect the streptozotocin degradation rate at all. However, streptozotocin levels in plasma isolated from rats decreased rapidly compared to those in neutral buffers.

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Studies on the Factors Influencing the Transformation in Escherichia with pBR322 DNA (Escherichia coli의 pBR322 DNA 형질전환에 관여하는 인자에 관한 연구)

  • Yoo, Han-sang;Mah, Jum-sool
    • Korean Journal of Veterinary Research
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    • v.24 no.1
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    • pp.40-49
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    • 1984
  • To investigate the factors influencing the artifical transformation in Escherichia coli, E. coli C600 was transformed by pBR322 DNA with tetracycline and ampicillin resistant gene purified by CsCl-Etbr equilibrium density gradient centrifugation from E.coli HB 101. The influencing factors in the transformation such as concentration of calcium chloride, time of ice incubation, temperature and time of heat shock, time of gene expression, effects of plasmid DNA concentration and adding time were examined in these experiments. The results obtained were as follows; 1. The highest transformation frequency was observed in the treatments of 100 mM $CaCl_2$ before heat shock and the treatment of $CaCl_2$ was essential step in the process of E. coli transformation. 2. The highest transformation frequency was observed in the treatment of heat shock at $42^{\circ}C$ for 4 min. or $37^{\circ}C$ for 6 min., but the prolonged heat shock resulted a decreased transformation frequency. 3. Treatments of ice incubation at $0^{\circ}C$ for 45 min. before heat stocks or at $0^{\circ}C$ for 30min. after heat shock resulted an increased transformation frequency. 4. There was a linear relationship between DNA concentration and transformation frequency at the concentration of $8{\times}10^3$ recipient cells. The highest transformation frequency reached in carte of 7 mcg of donor DNA, but above 1 mcg of DNA concentration, transformation frequency was not remarkably increased. Addition of donor DNA just after the treatment of $CaCl_2$ was the best. 5. The best condition of gene expression at $37^{\circ}C$ were 40min. for TC-resistant gene and 100min. for AP-resistant gene. TC-resistant gene was higher in the transformation frequency and faster in the gene expression time than AP-resistant gene. In these results, the best conditions for the transformation of E. coli C 600 with pBR322 DNA were: treatment with 100mM $CaCl_2$, ice incubation at $0^{\circ}C$ for 45 min, heat shock at $42^{\circ}C$ for 4 min., 30 min. of ice incubation and incubation at $37^{\circ}C$ for 100min. for gene expression in that order.

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Ruminal microbial responses in fermentation characteristics and dry matter degradability to TDN level of total mixed ration

  • Lee, Seung-Uk;Jo, Jin-Ho;Park, Sung-Kwon;Choi, Chang-Weon;Jeong, Jun;Chung, Ki-Young;Chang, Sun-Sik;Li, Xiang Zi;Choi, Seong-Ho
    • Korean Journal of Agricultural Science
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    • v.43 no.1
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    • pp.80-86
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    • 2016
  • An in vitro trial was conducted to examine the effects of total mixed rations (TMR) on fermentation characteristics and effective degradability (ED) by rumen microbes. Three TMR diets were growing period TMR (GR-TMR, 67% TDN), early fattening period TMR (EF-TMR, 75.4% TDN) and late fattening TMR (LF-TMR, 80% TDN). Three TMR diets (3 g of TMRs in each incubation bottles) was added to the mixed culture solution of stained rumen fluid with artificial saliva (1:1, v/v) and incubated anaerobically for 48 hours at $39^{\circ}C$. The pH in all incubation solutions tended to decrease up to 48h, but the opposite results were found in concentration of total gas production, ammonia-N and total VFA in all incubations.The total gas production (p<0.05) in LF-TMR was highest compared with those of other diets. Also, concentration of total VFA was tended to increase in LF-TMR compared with other TMR diets in all incubations. The EDDM in both EF-TMR and LF-TMR was tended to high compared with GR-TMR (p=0.100). In this in vitro trials, concentration of propionate in all incubation solution was not affected by increased concentration of TDN. The results of the present in vitro study indicate that TMR may provide more favorable condition for nutrient digestion both in the rumen.