The flavonoid, isokaempferide, has various biological activities such as hepatoprotective, antimicrobial and antiproliferative effect and is extracted from Amburana cearensis and Cirsium rivulare (Jacq.). Biotransformation is an alternative tool for the synthesis of value-added flavonoids with inexpensive substrates. Here, to synthesize isokaempferide from naringenin, two genes, PFLS and Rice O-mthyltransferae-9 were introduced in Escherichia coli. Although isokaempferide was successfully synthesized, the amount of biosynthesis was no high. In order to increase the yields of isokaempferide, S-adenosylmethionine (SAM) used as a methyl donor was increased by deleting MetJ, which is a transcriptional regulator related to SAM biosynthetic pathway. Next we optimized the cell concentration and substrate feed concentration with the engineered E. coli strain. Through these strategies, the biosynthesis of isokaempferide was increased up to 87 mg/L.
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.
The feasibility of skin penetration was studied for aspalatone (AM, acetylsalicylic acid maltol ester), a novel antithrombotic agent. In this studys hairless mouse dorsal skins were used as a model to select composition of vehicle and AM. Based on measurements of solubility and partition coefficient, the concentration of PC that showed the highest flux for AM across the hairless mouse skin was found to be 40%. The cumulative amount permeated at 48 h, however, appear inadequate, even when the PC concentration was employed. To identify a suitable absorption enhancer and its optimal concentration for AM, a number of absorption enhancers and a variety of concentration were screened for the increase in transdermal flux of AM. Amongst these, linoleic acid (LOA) at the concentration of 5% was found to have the largest enhancement factor (i.e., 132). However, a further increase in AM flux was not found in the fatty acid concentration greater than 5%, indicating the enhancement effect is in a bell-shaped currie. In a study of the effect of AM concentration on the permeation, there was no difference in the permeation rate between 0.5 and 1% for AM, below its saturated concentration. At the donor concentration of 2%, over the saturated condition, the flux of AM was markedly increased. A considerable degradation of AM was found during permeation studies, and the extent was correlated with protein concentrations in the epidermal and serosal extracts, and skin homogenates. In rat dorsal skins, the protein concentration decreased in the rank order of skin homogenate > serosal extract > epidermal extract. Estimated first order degradation rate constants were $6.15{\pm}0.14,{\;}0.57{\pm}0.02{\;}and{\;}0.011{\pm}{\;}0.004{\;}h^{-1}$ for skin homogenate, serosal extract and epidermal extract, respectively. Therefore, it appeared that AM was hydrolyzed to some extent into salicylmaltol by esterases in the dermal and subcutaneous tissues of skin. taken together, our data indicated that transdermal delivery of AM is feasible when the combination of PC and LOA is used as a vehicle. However, since AM is not metabolically stable, acceptable degradation inhibitors may be nervessary to fully realize the transdermal delivery of the drug.
The polymeric films containing drug and various excipients were fabricated using aqueous-based $Eudragit^{\circledR}$ RS 30D dispersions. The diffusional behaviors and mechanism of the fabricated polymeric film were investigated using Keshary-Chien diffusion cell. The melatonin was used as a model drug. The diffusion behaviors of drug through the fabricated polymeric films were highly dependent on drug concentration in donor part, polymer contents and drug concentration, and the types of plasticizers and solubilizers. The fabricated polymeric films containing excipients and solubilizers could be applied for the controlled release of poorly water-soluble drug and for the preparation of drug-containing latex films for topical or oral drug delivery.
The effect of protein supplementation, $O_2$ concentration and co-culture on the development of embryos produced by nuclear transfer using cultured cumulus cell was investigated. Recipient oocytes and cumulus cells were obtained from the ovaries of the slaughtered Hanwoo cows. Donor cumulus cells were cultured in Dulbecco's modified Eagle medium containing 10% fetal bovine serum at 5% $CO_2$ in air at $38.5^{\circ}C$. The 1 to 6 passages of cumulus cells were isolated and used as donor cells. The in vitro matured oocytes were enucleated and then the isolated donor cells were introduced. One $15{\mu}s$ pulse of 180 volts was applied to induce the fusion between karyoplast and cytoplast. The fused embryos were activated with $10{\mu}M$ calcium ionophore for 5 min and 2 mM 6-dimethylaminopurine for 3 h. To examine the effect of protein supplementation, nuclear transfer (NT) embryos were cultured in one of the following 4 treatments : 1) CR1aa + 3 mg/ml BSA for 7 days ; 2) CR1aa + 10% FBS for 7 days ; 3) CR1aa + 1.5 mg/ml BSA + 5% FBS for 7 days ; and 4) CR1aa + 3 mg/ml BSA for first 3 days and then CR1aa + 1.5 mg/ml BSA + 5% FBS for 4 days. Culture took place at 5% $CO_2$, 5% $O_2$ and 90% $N_2$ at $38.5^{\circ}C$. Although there were no significant differences in cleavage rate among different protein supplements, the rates of blastocyst formation were significantly different. When NT embryos were cultured in the medium supplemented with only BSA, they could develop to only morula not to blastocyst. However, when FBS was supplemented, NT embryos developed to blastocyst stage. In order to investigate the effect of $O_2$ concentration and co-culture, NT embryos were cultured in CR1aa + 1.5 mg/ml BSA + 5% FBS with or without cumulus cell co-culture at an atmosphere of 5% $CO_2$ in air (20% $O_2$) or 5% $CO_2$, 5% $O_2$, 90% $N_2$ (5% $O_2$) at $38.5^{\circ}C$ for 7 days. The percentage of blastocyst development was significantly higher when the NT embryos were cultured at an atmosphere of 5% $O_2$ than that of 20% $O_2$ (p<0.05). However, there was no significant difference between with and without cumulus cell co-culture at an atmosphere of 5% $O_2$ or 20% $O_2$. Fifty embryos were transferred to 25 recipients and 5 recipients were pregnant at 100 days. From 5 pregnant cows, only one cow was delivered of female twin. In conclusion, the embryos reconstructed by enucleation of metaphase II oocytes and introduction of the cycling and quiescent cumulus donor cells in Hanwoo had developmental potential to term after embryo transfer to recipient cows.
Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
/
2001.05b
/
pp.17-21
/
2001
We proposed that concentrations of cartier electron as well as ionized donor defects in nonstoichiometric ZnO are proportional to $P^{-1/2}_{O_2}$, whenever they ionizes singly or doubly, by employing the Fermi-Dirac (FD) statistics for ionization of the native thermal defects $Zn_i$ and $V_o$. The effect of acceptor defect, zinc vacancy $V_{Zn}$made by the Frenkel and Schottky disorder reactions, on carrier concentrations was discussed. By application of the FD statistics law to their ionization while the formation of defects is assumed governed by the mass-action law, the calculation results indicate; 1. ZnO shows n-type conductivity with $N_D>$N_A$ and majority concentration of $n{\propto}\;P^{-1/2}_{O_2}$ in a range of $P_{O_2}$, lower than a critical value. 2. As the concentration of acceptor $V_{Zn}$ increases proportional to $P^{1/2}_{O_{2}}$, ZnO made at extremely high $P_{O_{2}}$, can have p-type conductivity with majority concentration of p ${\propto}\;P^{-1/2}_{O_{2}}$. One may not, however, obtain p-type ZnO if the pressure for $N_{D}<$N_{A}$ is too high.
In order to Investigate the possibility as a simple technique of wastewater treatment for recirculating aquaculture system, the experiment by a biofilter unfit was carried out. The high and stable removal efficiency of nitrogen could be obtained by selecting the optimum recycle ratio and DO concentration. It was found that the proper combination of nitrifacation and denitrfication step in the reactor would be required for increasing the removal efficiency. The extent of nitrogen removal gradually decreased UO the rise of re- cycle ratio since the depression of denitrificatlon by the lack of hydrogen donor. The depression of nitrogen removal was overcome by increasing the CIN ratio In the wastewater. The extent of phosphorus removal was increased slightly with the increase of DO concentration and recycle ratio, but high removal efficiency was not observed. However, the extent of COD removal was not affected by recycle ratio and DO concentration and showed the stable removal of above 90%.
The purpose of this study is to characterize the iontophoretic transport of growth hormone releasing peptides (GHRP-6) through hairless mouse skin from aqueous solution. The effect of various factors, such as pH, poloarity, current profile, current density, current duration, ionic strength, drug concentration, and enhancer application was studied to obtain basic knowledge on the transport. We have also studied the stability of GHRP-6 in solution with/without current. The donor chamber was filled with phosphate buffer solution containing GHRP-6 and the receptor chamber was filled with phosphate buffer solution (pH 7.4). Ag/AgCl electrode was used for their stability and reversibility. At a predetermined time interval, sampling was made and the concentration of drug was analysed using HPLC system. The results showed that, compared to passive flux, the total amount of drug transported increased markedly by the application of anodal current. Cathodal flux was similar to passive flux. Flux increased with the current density, the duration of current application and drug concentration. The effect of enhancers on the flux was studied using hydrophilic (5% N-methyl pyrrolidone) and hydrophobic (5% propylene glycol monolaurate, 5% oleic acid) enhancers. Application of enhancer also increased the flux.
The two key variables of an Si solar cell, i.e., emitter (n-type window layer) and base (p-type substrate) doping levels or concentrations, are studied using Medici, a 2-dimensional semiconductor device simulation tool. The substrate is p-type and 150 ㎛ thick, the pn junction is 2 ㎛ from the front surface, and the cell is lit on the front surface. The doping concentration ranges from 1 × 1010 cm-3 to 1 × 1020 cm-3 for both emitter and base, resulting in a matrix of 11 by 11 or a total of 121 data points. With respect to increasing donor concentration (Nd) in the emitter, the open-circuit voltage (Voc) is little affected throughout, and the short-circuit current (Isc) is affected only at a very high levels of Nd, exceeding 1 × 1019 cm-3, dropping abruptly by about 12%, i.e., from Isc = 6.05 × 10-9 A·㎛-1, at Nd = 1 × 1019 cm-3 to Isc = 5.35 × 10-9 A·㎛-1 at Nd = 1 × 1020 cm-3, likely due to minority-carrier, or hole, recombination at the very high doping level. With respect to increasing acceptor concentration (Na) in the base, Isc is little affected throughout, but Voc increases steadily, i.e, from Voc = 0.29 V at Na = 1 × 1012 cm-3 to 0.69 V at Na = 1 × 1018 cm-3. On average, with an order increase in Na, Voc increases by about 0.07 V, likely due to narrowing of the depletion layer and lowering of the carrier recombination at the pn junction. At the maximum output power (Pmax), a peak value of 3.25 × 10-2 W·cm-2 or 32.5 mW·cm-2 is observed at the doping combination of Nd = 1 × 1019 cm-3, a level at which Si is degenerate (being metal-like), and Na = 1 × 1017 cm-3, and minimum values of near zero are observed at very low levels of Nd ≤ 1 × 1013 cm-3. This wide variation in Pmax, even within a given kind of solar cell, indicates that selecting an optimal combination of donor and acceptor doping concentrations is likely most important in solar cell engineering.
For the production of $2-O-{\alpha}-D-glucopyranosyl-L-ascorbic$ acid (ascorbic acid-2-g1ucoside, AA-2G) from ascorbic acid, the usability of spinach seed as the source of ${\alpha}-glucosidase$ having transglucosylation activity was studied. The optimum conditions for the production of AA-2G from ascorbic acid and glucose donor were investigated by using crude extract of Spinachia oleracea L. Woosung, the selected source of enzyme. The production of AA-2G was the highest with 1.053 mM when spinach seeds were grown for 2 days after germination. Maltose was the most effective glucose donor, and the optimum concentration of ascorbic acid and maltose were 175 mM and 225 mM, respectively. The optimum concentration of ${\alpha}-glucosidase$ was 60 units. The most effective buffer was sodium acetate and its optimum concentration was 175 mM. The optimum pH and temperature were 5.0 and $65^{\circ}C$, respectively. Under the optimum condition, 2.14 mM of AA-2G was produced from ascorbic acid after 50 minutes of reaction.
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