Purpose: It has been reported that all-trans retinoic acid (ATRA) can inhibit glioma growing in vitro. However, clinical trials with ATRA alone in gliomas revealed modest results. ATRA has been shown to increase radiosensitivity in other tumor types, so combining radiation and ATRA would be one of alternatives to increase therapeutic efficacy in malignant gliomas. Thus, we intended to know the role of catalase, which is induced by ATRA, for radiosensitivity if radiation-reduced reactive oxygen species (ROS) is removed by catalase, the effect of radiation will be reduced. Materials and Methods: A rat glioma cell line (36B10) was used for this study. The change of catalase activity and radiosensitivity by ATRA, with or without 3-amino-1, 2, 4-triazole (ATZ), a chemical inhibitor of catalase were measured. Catalase activity was measured by the decomposition of $H_2O_2$ spectrophotometrically Radiosensitivity was measured with clonogenic assay. Also ROS was measured using a 2, 7-dichlorofluorescein diacetate spectrophotometrically. Results: When 36B10 cells were exposed to 10, 25 and $50{\mu}M$ of ATRA for 48 h, the expression of catalase activity were increased with increasing concentration and incubation time of ATRA. Catalase activity was decreased with increasing the concentration of AT (1, $10{\mu}M$) dose-dependently. ROS was increased with ATRA and it was augmented with the combination of ATRA and radiation. ATZ decreased ROS production and increased cell survival in combination of ATRA and radiation despite the reduction of catalase. Conclusion: The increase of ROS is one of the reasons for the increased radiosensitivity in combination with ATRA. The catalase that is induced by ATRA doesn't decrease ROS production and radiosensitivity.
Green tides, which was mainly caused by Ulva spp., have been increasing in severity and frequency globally, and have negatively affected on marine ecosystems. This study was conducted to investigate effects of various physical and chemical factors on the death of Ulva australis (ULAUS) and to consider a practical measures useful for alleviating Ulva bloom. Soaking of ULAUS thalli in pure water for 8 hr didn't induce a death, but incubation in 1.0-1.5% salinity for 7 d inhibited sporulation by about 70%. Desiccation gave rise to a serious damage when more than 40-50% of initial fresh weight was lost. ULAUS growth was sensitive to temperature and seriously inhibited from more than $30^{\circ}C$. At $35^{\circ}C$, $40^{\circ}C$, $45^{\circ}C$ and $50^{\circ}C$, treatment time required for 90-95% death of ULAUS thalli was 1 d, 10 min, 30 sec, and 1 sec, repectively. ULAUS growth was seriously inhibited at lower than pH 6.0 and completely dead at pH 4.0. Several compounds for ULAUS control was selected and the chemcals causing a rapid death were oxidants such as hydrogen peroxide and sodium percarbonate. Taken together, our results suggest that low salinities, dryness, pH, high temp. and compounds could be selected for Ulva bloom control, and high temperature and compounds seems to be useful for a development of practical control methods.
Background : Potassium channel opener (K-opener) opens ATP-sensitive K'-channel located at cell membrane and induces potassium efflux from cytosol, resulting in intracellular hyperpolarization. Newly synthesized K-opener is currently examined for pharmacologic potency by means of rubidium release test from smooth muscle strip pre-incubated with Rb-86. Since in-vivo behavior of thallium is similar to that of rubidium, we hypothesized that K-opener can alter T1-201 kinetics in vivo. Purpose : This study was prepared to investigate the effects of pinacidil (one of potent K-openers) on the T1-201 uptake and clearance in cultured myocyte, and in-vivo biodistribution in mice. Methods : Spontaneous contracting myocytes were prepared to imitate in-vivo condition from 20 hearts of 3-5 days old Sprague-Dawley rat and cultured for 3-5 days before use ($5{\times}10^5$ cells/ml). Pinacidil was dissolved in 10% DMSO solution at a final concentration of 100nM or l0uM and was co-incubated with T1-201 in HBSS buffer for 20-min to evaluate its effect on cellular T1-uptake, or challenged to cell preparation pre-incubated with T1-201 for washout study. Two, 40 or $100{\mu}g$ of pinacidil was injected intravenously into ICR mice at 10 min after $5{\mu}Ci$ T1-201 injection, and organ uptake and whole body retention rate were measured at different time points. Results : Co-incubation of pinacidil with T1-201 resulted in a decrease in T1-201 uptake into cultured myocyte by 1.6 to 2.5 times, depending on pinacidil concentration and activity of T1-201 used. Pinacidil enhanced T1-201 washout by 1.6-3.1 times from myocyte preparations pre-incubated with T1-201. Pinacidil treatment appears to be resulted in mild decreases in blood and liver activity in normal mice, in contrast, renal and cardiac uptake were mildly decreased in a dose dependent manner. Whole body retention ratios of T1-201 were lower at 24 hour after injection with $100{\mu}g$ of pinacidil than control. Conclusion : These results suggest that treatment with K-opener may affect the interpretation of T1-201 myocardial images, due to decreasing thallium accumulation and enhancing washout from myocardium.
The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
/
v.4
no.1
/
pp.33-39
/
1999
In order to understand the importance of tidal action and $NH_4{^+}$ -nitrification in the removal of dissolved oxygen (DO) and $NH_4{^+}$, concentrations of DO, $NH_4{^+}$, $NO_2{^-}$ and $NO_3{^-}$ were measured with time for water samples collected at different tidal state in the eutrophic macrotidal Han River estuary. Field measurements indicated that most environmental parameters, except for the water temperature and DO concentration, were tightly controlled by the eutrophic freshwater runoff and large-scale tidal action. Dark incubation of the water sample at $25^{\circ}C$ showed that the removal rates of DO and $NH_4{^+}$ in high tide sample were 2.76 ${\mu}M\;O_2\;d^{-1}$ and 1.76 ${\mu}M\;N\;d^{-1}$ respectively, and increased to 5.66 ${\mu}M\;O_2\;d^{-1}$ and 3.36 ${\mu}M\;N\;d^{-1}$ respectively, in low tide sample. These changes indicated that microbial degradation and uptake of organic matter and inorganic nutrients were more active during low tide. $NH_4{^+}$-nitrification responsible for total DO removal in low tide (23.81%) and $NH_4{^+}$ turnover rates due to $NH_4{^+}$-nitrification in low tide (0.18 $d^{-1}$) were approximately 3.7 times and 3 times, respectively, higher than those in high tide. These results indicated that $NH_4{^+}$ -nitrifying bacteria introduced into the Han River estuary during low tide played a significant role in the removal of DO and $NH_4{^+}$. The decreasing removal rates in DO and $NH_4{^+}$ with the increasing tidal level seemed to be associated with the salinity impact on the halophobic freshwater $NH_4{^+}$-nitrifying bacteria. The results implied that anthropogenic $NH_4{^+}$ sources should be treated prior to the freshwater runoff into the estuary for the effective control of $NH_4{^+}$ in the Han River estuary. These results also suggest that parallel ecological studies on the chemoautotrophic nitrifying bacteria are essential for the elucidation of nitrogen cycles in the eutrophic Han River estuary.
Kim, Sang-Woo;Park, Kyung-Min;Ha, Jae-Uk;Lee, Jae-Hwan;Chang, Pahn-Shick
Korean Journal of Food Science and Technology
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v.41
no.2
/
pp.173-178
/
2009
Epidemiological studies showed that high trans-fat consumption is closely associated with getting the risks of cardiovascular disease. The objective of this study was to produce trans-free fat through lipase-catalyzed interesterification, as a substitute for the cream margarine commonly used in industry. Optimum conditions for interesterification in a packed bed enzyme bioreactor (PBEB) were determined using response surface methodology (RSM) based on central composite design. Three kinds of reaction variables were chosen, such as substrate flow rate (0.4-1.2 mL/min), reaction temperature (60-70$^{\circ}C$), and ratio of fully hydrogenated canola oil (FHCO, 35-45%) to evaluate their effects on the degree of interesterification. Optimum conditions from the standpoint of solid fat content (SFC) were found to be as follows: 0.4 mL/min flow rate, 64.7$^{\circ}C$ reaction temperate, and 42.8% (w/w) ratio of FHCO, respectively. The half-life of immobilized lipase in PBEB with two stages at 60$^{\circ}C$ ($1^{st}$ stage) and 55$^{\circ}C$ ($2^{nd}$ stage) was about more than 30 days as estimated by extrapolating the incubation time course of tristearoyl glycerol (TS) conversion, whereas the half-life of the enzyme in PBEB with single stage at 65$^{\circ}C$ was only about 15 days. Finally, the results from SFC analysis suggest that trans-free fat produced in this study seems to be a suitable substitute for the cream margarine commonly used in industry.
Kim, Min-Jung;Kim, Ji-Young;Leec, Seung-Jae;Yoon, Yong-Dal;Cho, Dong-Jae;Kim, Hae-Kwon
Development and Reproduction
/
v.5
no.1
/
pp.23-33
/
2001
When mammalian oocytes ovulate into the oviduct, associating follicular fluid components are exposed to the oviductal environment, possibly resulting in the mutual interaction between fillicu1ar and oviductal fluids. In the Present study, we have demonstrated for the first time that components of fallicular fluid could be modified by the oviductal fluid. Gelatin zymographic analyses of human follicular fluid (hFF) obtained from IVF patients showed consistently the presence of 110 kDa gelatinase (GA110) in addition to many bands among which 62 kDa gelatinase was predominant. Addition of EDTA or phenanfhroline to the gelatinase substrate buffer during gel incubation abolished GA110 band whereas phenylmethylsulffnyl fluoride (PMSF) did not. In contrast, bovine oviductal fluid(bOF) exhibited only 62 kDa gelatinase. Surprisingly, when bOF was added to hFF in 1:1 ratio and then the mixture was incubated for 3 h at 37$^{\circ}$C, GA110 of hFF disappeared. Disappearance of GA110 by bOF was observed even within 30 min after mixing with hFF. Addition of aminophenylmercuric acetate (APMA) to hFF also abolished enzymatic activity of GA110 but increased the activityof 62 kDa gelatinase. However, APMA abolished many other gelatinases as well unlike bOF. Interestingly, treatment of hFF with EDTA for 3 h remarkably increased the enzymatic activity of GA110 but not that of other gelatinases. Addition of phenanthroline, PMSF or soybean trypsin inhibitor (SBTI) did not affect overall gelatinase activities. Again, addition of bOF to the hFF pretreated with any of the above proteinase inhibitors abolished the appearance of GA110. Human serum also showed GAI 10 of which activity was greatlyenhanced by EDTA treatment. Similar to hFF, serum GA110 also disappeared by the addition of bOF. Human granulosa cell homogenate did not reveal any appreciable gelatinase activity except 92 kDa gelatinase. Anti-human gelatinase A antibody reacted with 62 kDa gelatinase of hFF. Based upon these results, it is concluded that bOF could selectively degrade an isoform of gelatinase A present in hFF and human serum.
This study was undertaken to investigate the effects of ruminally protected amino acids (Methionine and Lysine) on in vitro ruminal parameters, and in vivo milk yield and milk composition in mid-lactating cows. In the first in vitro experiment, there were no statistical significances between treatments in ruminal pH and dry matter digestibility during various incubation times. In the second in vivo experiment, milk yield decreased by 11.92% in control and 5.68% in the treatment respectively, but decrease rate of milk yield in the treatment was lower than control. Milk yields naturally decreased as time goes by since the DIMs(Days in milk) of the cows in experiment were in mid-lactation period. 4% FCM(Fat corrected milk) and milk protein yields also, respectively, decreased by 11.25% and 11.09% in control and 6.16% and 5.47% in the treatment as compared with the intial. Milk protein and milk fat production were higher in the treatment(0.90kg, 1.10kg) than those of control(0.66kg, 0.79kg). Milk fat content significantly increased with supplementing protected amino acids as compared to control(P<0.05). From the above results, protected amino acids were positively utilized in the performances of mid-lactating cows without inhibiting rumen fermentation. Further investigation is suggested for essential amino acid composition and intestinal digestion rate out of rumen bypass protein in dietary protein to be estimated.
DNA transfection is a powerful tool for studying gene functions. The $Ca^{2+}$-phosphate precipitation remains one of the most popular and cost-effective transfection techniques. Mature neurons are more resistant to transfection than young ones and most other cell types, and easy to die if microenvironment changes. Here, we report a transfection protocol for mature neurons. The critical modifications are inclusion of glial cells in culture and careful control of $Ca^{2+}$-phosphate precipitation under microscope. Cerebral glial cells were grown until ${\sim}70-80%$ confluence in DMEM/10% horse serum, which was thereafter replaced with serum-free Neurobasal/Ara-C, and 319 hippocampal neurons were plated onto the glial layer Formation of fine $DNA/Ca^{2+}$-phosphate precipitates was induced using Clontech $CalPhos^{TM}$ Mammalian Transfection Kit, and the size ($0.5-1\;{\mu}m$ in diameter) and density(about 10 particles/$100\;{\mu}m^2$) were carefully controlled by the time of incubation in the medium. This modified protocol can be reliably applied for transfection of mature neurons that are maintained longer than two weeks in vitro, resulting in 10-15 healthy transfected neurons per a well of 24-well plates. The efficacy of the protocol was verified by punctate expression of $pEGFP-CaMKII{\alpha}$, a synaptic protein, and diffuse expression of pDsRed2. Our protocol provides a reliable method for transfection of mature neurons in vitro.
Jeong, Su-Ji;Yang, Hee-Jong;Ryu, Myeong Seon;Seo, Ji Won;Jeong, Seong-Yeop;Jeong, Do-Youn
Journal of Life Science
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v.28
no.5
/
pp.577-586
/
2018
We recently reported the potential probiotic properties of Lactobacillus brevis SBB07 isolated from blueberries. The present study investigates the effect of culture conditions such as temperature, initial pH, culture time, and medium constituent for industrial application. The ingredients of the medium to improve cell growth were selected by Plackett-Burman design (PBD) and central composite design (CCD) within a desirable range. The PBD was applied with 19 factors: seven carbon sources, six nitrogen sources, and six microelements. Protease peptone, corn steep powder (CSP), and yeast extract were found to be significant factors for the growth of SBB07. The CCD was then applied with three variables found from the PBD at five levels, and the optimum values were decided for the three variables: protease peptone, CSP, and yeast extract. In the case of the growth of SBB07, the proposed optimal media contained 2.0% protease peptone, 2.5% CSP, and 2.0% yeast extract, and the maximum dried-cell weight was predicted to be 2.93963 g/l. By the model verification, it was confirmed that the predicted and actual results are similar. Finally, the study investigated the effects of incubation temperature and initial pH at the optimized medium. It was confirmed that the dried-cell weight increased from $2.2933{\pm}0.0601g/l$ to $3.85{\pm}0.0265g/l$ when compared to the basal medium at $37^{\circ}C$ and initial pH 8.0. Establishing the optimal culture condition for SBB07 provides good potential for applications in probiotics and can serve as the foundation for the industrialization of materials.
Slow-release fertilizers (SRF) have been used to reduce nutrient loss through increasing fertilizer efficiency and to save labor. Several SRFs were developed for rice plant in Korea, but there is few for horticultural crop plants. Two slow-release complex fertilizers, 100T and 150T, which made for controlling nitrogen release time up to 100 and 150 days, respectively, were selected for the incubation test cto evaluate nitrogen (N) release rate in soil. The N of urea selected as the control was completely released within a week after application. Sixty three and 53% of total N were released from 110T and 150T of slow release fertilizers within 8th weeks after application, respectively. For pepper cultivation CF110 and CF150, new slow-release complex fertilizer, were made of mixing 40% of conventional fertilizer and 60% of 110T and 150T, respectively, based on the amount of recommended fertilizer for pepper cultivation $(N-P_2O_5-K_2O=190-112-149\;kg\;ha^{-1})$, and were totally applied before pepper transplanting in the field as the basal fertilizer. Inorganic N $(NH_4^+-N+NO_3^--N)$ concentration in soil was higher in the CF110 treatment than in the control (NPK) at all period of pepper cultivation. In the CF150 treatment concentration of inorganic N in soil was low compared to control up to 8th weeks after transplanting. However, there was no difference in plant height and nutrient content of pepper leave between CF110 treatment and the control. In comparison, plant height was significantly lower in CF150 than the control and CF110 treatments. Around 4% of fresh pepper yield was increased in CF110 compared to the control, but it was decreased to about 2% by CF150 treatment. Conclusively, CF110 form could be recommended as a slow release fertilizer for pepper cultivation.
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