• Tuberculosis and Respiratory Diseases
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• v.44 no.3
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• pp.547-558
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• 1997

Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majorities of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to hoot. In previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF. Understanding the mechanisms of TNF-resistance in TNF-$\alpha$ gene transfected cancer cells would be an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of new protective protein synthesis in the acquired resistance to TNF of TNF-$\alpha$ gene transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164, a murine fibrosarcoma cell line, using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF gene transfected cells(WEHI164-TNF) and the changes of TNF sensitivities after treatments with actinomycin D(transcription inhibitor) and cycloheximide ( translation inhibitor). Results : WEHI164 which was sensitive to TNF became resistant to TNF after being transfected with TNF-$\alpha$ gene and the resistance to TNF was partially reversed after treatment with actinomycin D, but not with cycloheximide. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ gene transfection may be associated with synthesis of some protective proteins.

• KSBB Journal
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• v.23 no.6
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• pp.513-518
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• 2008

This paper described the extraction/purification of $\beta$-carotene from recombinant E.coli and evaluation of anti-wrinkle activity of purified $\beta$-carotene. No significant differences in extraction yields were observed when hexane or isobutyl acetate was used. However, extraction from wet-cell cake resulted in 2-fold higher amount of $\beta$-carotene than that from dry cells. Disruption of 5 g-wet cells by ultrasonic homogenizer, acetone dehydration, extraction with isobutyl acetate resulted in 36 mg of $\beta$-carotene corresponding to 61.2% of recovery. The formation and separation of $\beta$-carotene crystal improved the purity. 633 mg of $\beta$-carotene crystal with 93% purity was obtained from 223 g/L of wet-cell cake harvested from 2.5-L fed-batch culture broth. The cultures of normal human primary fibroblast were performed to investigate the effect of $\beta$-carotene on cytotoxicity as MTT assay and anti-wrinkle activity as collagen synthesis assays. $1.7{\mu}M$ of $\beta$-carotene was found to be optimal concentration at which 1.4-fold higher amount of collagen was synthesized than that in absence of $\beta$-carotene. This indicates that highly purified $\beta$-carotene can be obtained from recombinant E.coli by applying simple method with less toxic solvent and can be used in functional cosmetics as anti-wrinkle agent.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.19 no.1
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• pp.12-19
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• 2019

Purpose: Isolated methylmalonic acidemia (MMA) is an autosomal recessive inherited disorder of propionate metabolism. There are two subtypes of MMUT gene defects. $Mut^0$ represents complete loss of methylmalonyl-CoA mutase (MCM) activity while mut- is associated with residual MCM activity, which can be stimulated by hydroxocobalamin (OHCbl) supplementation. The objective of this study is to investigate cobalamin responsiveness and mutations present in Korean MMA population. Methods: We evaluated 10 MMA patients using somatic cell complementation analysis on their fibroblasts to measure MCM activity and vitamin B12 responsiveness for the optimal treatment. MMUT gene was sequenced to identify the MMA mutations. Results: For all patients, the incorporation of $[^{14}C]-propionate$ was low, and there was no response to OHCbl. The incorporation of $[^{14}C]-methyltetrahydrofolate$ and $[^{57}Co]-CNCbl$ fell within the normal range. There was adequate synthesis of methylcobalamin while the synthesis of adenosylcobalamin was low. The complementation analysis showed all patients were $mut^0$. The sequence analysis identified 12 different MMUT mutations, including 2 novel mutations, p.Gln267Ter and p.Ile697Phe, were identified. All the patients in this study had neonatal onset of symptoms, belonged to $mut^0$ complementation class, and as a result, showed no cobalamin responsiveness. Conclusion: No Korean MMA patient showed cobalamin responsiveness.

• Journal of the Korean Applied Science and Technology
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• v.40 no.3
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• pp.534-546
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• 2023

In this study, we investigated the antioxidant and anti-winkle activity in human fibroblast cell (CCD-986sk) of Persimmon Leaves (PL) as a cosmetic ingredient. As a result of investigating antioxidant activity through electron-donating ability and ABTS⁺ radical scavenging assay, the PL showed concentration-dependent antioxidant activity similar to ascorbic acid, a control group, at a concentration of 1,000 ㎍/ml. As a result of investigating the anti-wrinkle effect through elastase inhibition and collagenase inhibition assay, the PL showed concentration-dependent antioxidant activity similar to epigallocatechin gallate, a control group, at a concentration of 1,000 ㎍/ml. As a result of measuring the synthesis rate of pro-collagen type I and the inhibition rate of MMP-1 in UVB-induced CCD-986sk cells, the control group EGCG showed a 90.2% pro-collagen synthesis rate at 20 ㎍/ml and PL showed an 88.5% synthesis rate at 30 ㎍/ml. In addition, the inhibition rate of MMP-1 of 33.0% and 40.8% were confirmed in EGCG 20 ㎍/ml and PL 30 ㎍/ml, respectively. As a result of measuring the protein expression of pro-collagen type I and MMP-1 in the PL through western blot, it was confirmed that the protein expression of pro-collagen type I increased, and MMP-1 decreased when the PL was treated together compared to the UVB alone group. According to the above experimental results, it is expected to be used as a natural product material for cosmetics by confirming that the PL prevent photoaging caused by UVB stimulation and have antioxidant and anti-wrinkle effects.

• Journal of Life Science
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• v.28 no.7
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• pp.841-848
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• 2018

The flavonoid content and antioxidant effects of extracts from Stachys sieboldii Miq. and Lycopus lucidus Turcz were compared. The flavonoid content of the acetone + methylene chloride (A+M) extract of L. lucidus Turcz was 233.2 mg/g, suggesting that the extract was greater than that of S. sieboldii Miq. In the DPPH assay and the A+M and methanol (MeOH) extracts from L. lucidus Turcz had greater scavenging effects than those of S. sieboldii Miq. (p<0.05). The A+M extract from L. lucidus Turcz (0.5 mg/ml concentration) had an 82% scavenging effect in the DPPH assay. In the ABTS assay, A+M extracts from both S. sieboldii Miq. and L. lucidus Turcz (0.5 mg/ml concentration) had scavenging effects of 90% and 88%, respectively (p<0.05), suggesting that both A+M extracts had greater scavenging effects than those of both MeOH extracts. In a 120 min ROS production assay, all tested extracts dose-dependently decreased the cellular ROS production that was induced by $H_2O_2$, as compared to those produced by exposure to the extract-free control. The A+M extracts from both S. sieboldii Miq. and L. lucidus Turcz had greater inhibitory effects on cellular ROS production than those of both MeOH extracts at all concentrations tested. Treatment with the A+M extracts from S. sieboldii Miq. and L. lucidus Turcz (0.25 mg/ml concentration) inhibited the cellular ROS production by 60% and 86%, respectively. These results suggest that the A+M extracts of Stachys sieboldii Miq. and L. lucidus Turcz inhibit cellular oxidation and may contain valuable bioactive compounds, such as flavonoids.

• Reproductive and Developmental Biology
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• v.29 no.1
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• pp.1-7
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• 2005

Telomeres consisting of (TTAGGG)n tandem repeat DNA sequences and associated proteins are essential for chromosome stability and related with cell senescence, apoptosis and cancer. The telomerase is a ribonucleoprotein which act as a template for the synthesis of telomeric DNA. This study was carried out to identify the distribution of telomeres on mouse chromosomes and also to analyze the amount of telomeres and telomerase activity of mouse embryos at early embryonic stages. Germ cells and early embryos from 1 cell to blastocyst stage were analyzed. The amount of telomeres was analyzed by quantitative fluorescence in situ hybridization technique(Q-FISH) using a human telomeric DNA probe, and telomerase activity was measured by telomeric repeat amplification protocol assay(TRAP). In results, the telomeres on mouse chromosomes were distributed at the ends of all autosomes and sex chromosomes. Although the quantity of telomeres varied among chromosomes, most of chromosomes had higher amount in q-arm telomeres than in p-arm telomeres. The results of Q-FISH indicated that the relative amount of telomeres of mouse embryos in each embryonic stage was approximately the same except the higher amount in blastocysts. Using TRAP assay on mouse embryos, telomerase activity was detected in all preimplantation stages from mature oocytes to blastocysts. Especially the telomerase activity was significantly increased at the morula and blastocyst stage. In conclusion, there may be a close association between the amount of telomeres and telomerase activity in early embryonic stages, and analysis of telomere quantity and telomerase activity on early development will be helpful for the investigation of embryogenesis and embryonic cell differentiation in mice.

• Journal of the Korean Wood Science and Technology
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• v.45 no.1
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• pp.126-138
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• 2017

This study was conducted to investigate the potential of chicken feather (CF), which is a by-product in poultry industry, as a raw material of wood adhesives. For the purpose, adhesive resins were formulated with NaOH- and $H_2SO_4$-hydrolyzed CF as well as crosslinkers, and then the properties and water resistance of the adhesive resins against hot water were measured. CF was made of mainly keratin-type protein, and no or very low content of heavy metals was detected. Hydrolysis rate of CF increased as NaOH concentration in hydrolysis solutions increased. However, in order to minimize the loss of adhesive property of protein itself by the severe hydrolysis of CF and to seek its proper hydrolysis conditions, NaOH concentrations in hydrolysis solution determined to adjust to 5%, 7.5% and 10%. In the NaOH-hydrolyzed CF, $H_2SO_4$-hydrolyzed CF as a hardener and crosslinker were added to formulate CF-based adhesive resins. Solid content of the resins ranged from 28.3% to 44.8% depending on hydrolysis conditions and type of crosslinker. Viscosity of the resins at $25^{\circ}C$ was very high. However, when the temperature of the resins was increased to $50^{\circ}C$, the viscosity decreased greatly and thus the resins could be applied as a sprayable resin. Retention rate measured to evaluate the water resistance of adhesive resins was the highest in the cured resin formulated with 5% NaOH-hydrolyzed CF and 5% $H_2SO_4$-hydrolyzed CF of 10% based on the solid weight as a hardener. Retention rate depending on crosslinkers added into adhesive resins was the highest phenol-formaldehyde (PF) followed by melamine-urea-formaldehyde (MUF) and formalin. The retention rate of CF-based adhesives formulated with 5% NaOH-hydrolyzed CF, PF and $H_2SO_4$-hydrolyzed CF of 10% and over did not differ statistically from that of commercial MUF resins. All of CF-based adhesives formulated with PF as a crosslinker and one with 5% NaOH-hydrolyzed CF of 55%, 5% $H_2SO_4$-hydrolyzed CF of 15%, and MUF of 30% on the basis of solid weight could be substituted for commercial urea-formaldehyde resins, From the results, CF can be used as a raw material of wood adhesives if hydrolyzed in proper conditions.

• Clean Technology
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• v.22 no.1
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• pp.16-28
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• 2016

Textile industry is considered as one of the most polluting sectors in terms of effluent composition and volume of discharge. It is well known that the effluents from textile dying industry contain not only chromatic substances but also large amounts of organic compounds and insolubles. The azo dyes generate huge amount of pollutions among many types of pigments. In general, the electrochemical treatments, separating colors and organic materials by oxidation and reduction on electrode surfaces, are regarded as simpler and faster processes for removal of pollutants compared to other wastewater treatments. In this paper the electrochemical degradation characteristics of dye wastewater containing CI Direct Blue 15 were analyzed. The experiments were performed with various anode materials, such as RuO₂/Ti, PtO₂/Ti, IrO₂/Ti and graphite, with stainless steel for cathode. The optimal anode material was located by changing operating conditions like electrolyte concentration, current density, reaction temperature and initial pH. The degradation efficiency of dye wastewater increased in proportion to the electrolyte concentration and the current density for all anode materials, while the temperature effect was dependent on the kind. The performance orders of anode materials were RuO₂/Ti > PtO₂/Ti > IrO₂/Ti > graphite in acid condition and RuO₂/Ti > IrO₂/Ti > PtO₂/Ti > graphite in neutral and basic conditions. As a result, RuO₂/Ti demonstrated the best performance as an anode material for the electrochemical treatment of dye wastewater.

• Journal of Life Science
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• v.30 no.2
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• pp.147-155
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• 2020

In this study, the antioxidant and cytotoxic effects and the flavonoid contents of leaf extracts from Stachys sieboldii Miq. and Lycopus lucidus Turcz. were compared. The flavonoid contents of the acetone + methylene chloride (A+M) and methanol (MeOH) extracts of L. lucidus Turcz. leaves were 55.7 and 233.2 mg/g, respectively. In a DPPH assay, A+M and MeOH extracts from L. lucidus Turcz leaves had a greater scavenging effect than those of S. sieboldii Miq. leaves (p<0.05). In an ABTS assay, MeOH extracts from S. sieboldii Miq. and L. lucidus Turcz (0.5 mg/ml concentration) leaves had scavenging effects of 85% and 91%, respectively (p<0.05), suggesting that both of the MeOH extracts had greater scavenging effects than both A+M extracts. In a 120 min ROS production assay, all tested extracts decreased the cellular ROS production induced by H₂O₂ compared to that produced by exposure to the extract-free control. The MeOH extract from L. lucidus Turcz leaves had a greater inhibitory effect on cellular ROS production (p<0.05). Treatment with A+M and MeOH extracts from both S. sieboldii Miq. and L. lucidus Turcz. leaves showed a dose-dependent increased cytotoxicity against the growth of AGS, HT-29 cancer cells, and HT-1080 (p<0.05). Both A+M extracts had a greater inhibitory effect on the growth of all cancer cells than both MeOH extracts. These results suggest that the MeOH extract of L. lucidus Turcz. leaves is effective in scavenging free radicals and inhibiting cellular oxidation, while the A+M extract inhibits proliferation of three types of cancer cell.

• Applied Biological Chemistry
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• v.18 no.4
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• pp.203-218
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• 1975

In order to investigate the compositional change oil composts during the growing of cultivated mushroom (Agaricus bisporus), composts and mushrooms during the period of filling to ending under commercial conditions were subjected to chemical analyses. The results are summarized as follows and the mechanism of composting for mushroom cultivation was proposed. 1) The temperature change of growing bed and room was observed and the yield of mushroom for each cropping time was recorded to get $15.6kg/m^2$ in total crops. 2) Composts after filling showed pH 8.2 which dropped to 6.4 after casing and continued so up to ending. 3) On the dry weight basis of composts, crude ash increased whereas total nitrogen, ether extract and crude fibre decreased gradually to bring about the lowering of organic matter. 4) Total nitrogen of composts decreased gradually and more insoluble nitrogen was lost than soluble nitrogen. The C/N ratio of composts was initially 21 which was gradually lowered to 16. 5) The losses of ${\alpha}-cellulose$, pentosan and lignin in composts were 87%, 75%, and 60%, respectively, in which ${\alpha}-cellulose$ decreased markedly after casing. 6) Free reducing sugars of composts increased continuously. Gradually increased free amino acids till second cropping decreased again thereafter. Composts at the filling stage contained alanine, glutamic acid, glycine and serine in which glycine decreased markedly whereas proline increased remarkably upon mushroom cultivation. 7) Among minerals of composts, phosphorus and zinc tended to decrease, potassium and copper tended to increase anti sodium showed no marked change. 8) In comparison of mushrooms from different cropping time with respect to proximate composition, minerals, free reducing sugars and amino acids, no marked difference was observed. However, a little higher values were observed in crude fat, free reducing sugars and sodium content for early crops and in free amino acids and phosphorus content for late crops. Twelve free amino acids including alanine, serine, threonine, and glutamic acid were detected in the cultivated mushroom. 9) According to above experimental results, it was possible to support the mechanism of compositing that the formation of ammonia and decomposition of carbohydrates by mesophiles are followed by protein biosynthesis, formation of microbial bodies and nitrogen-rich lignin humus complex by thermophiles, thus supplying necessary nutrients for mushroom growth, along with residual carbohydrates.