• Title/Summary/Keyword: Cell Production

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Viability of Endothelial Cells in Preserved Human Saphenous Vein Allografts (보존된 사람 동종 복재정맥 이식편혈관 내피세포의 생활성에 관한 연구)

  • 지현근;김용진
    • Journal of Chest Surgery
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    • v.36 no.4
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    • pp.229-241
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    • 2003
  • Background:Autogenous vein is the preferred vascular graft for patients who require coronary artery bypass surgery or peripheral arterial bypass surgery. When an autogenous vein is not available, an allograft saphenous vein can be used as an alternative conduit. Although arterial homograft has been under investigation since the beginning of this century, the viability of endothelial cells and the optimum mode of storage for the venous and arterial allografts is controversial. In addition, with the recently gained knowledge of vascular endothelial functions, such as the production of nitric oxide or thrombomodulin, the viability and antigenicity of endothelial cells are being studied again. The purpose of this study was to evaluate the viability of endothelial cells in the preserved human saphenous veins. Material and Method: The veins were stored in a $4^{\circ}C$ RPMI (Roswell Park Memorial Institute) 1640 solution including 10% fetal calf serum, for one, three, five, seven or fourteen days. After the completion of the storage period, the veins were divided into two groups: Group I: studied immediately at $4^{\circ}C$ (cold) storage (I-1, I-3, I-5, I-7, I-14), and Group II: studied after storage at $-196^{\circ}C$ liquid nitrogen tank (cryopreservation) in an RPMI 1640 solution containing 10% DMSO for two weeks (II-1, II-3, II-5, II-7, II-14). Light microscopy and scanning electron microscopy (SEM), frypan blue exclusion testing, and thrombomodulin immunohistochemistry were performed. Result: In a morphometric study using SEM, there was statistically significant increase in Gundry Score in Groups I-7, I-14, II-5, II-7, and II-14 and showed cellular destruction (p<0.05). In the thrombomodulin immunohistochemistry study, there was reactivity in Groups I-1, I-3, and I-5, but the cryopreserved group revealed decreased reactivity (p<0.05). The trypan blue exclusion testing also showed superior viability in cold storage Group I. Conclusion: Venous allografts preserved in a $4^{\circ}C$ RPMI 1640 solution showed well preserved endothelial cellular integrity and thrombomodulin expression at up to seven days of preservation. Although cryopreservation of venous allografts stored in 10% DMSO -RPMI 1640 solution maintained the endothelial cellular structure on SEM, immunohistochemistry from the thrombomodulin and trypan blue exclusion testing showed decreased viability, It remains to be seen whether the decreased thrombomodulin reactivity could be restored, and what the nature to the relationship is between thrombomodulin and long-term patency of allografts.

Enhancement of Immune Activities of Peptides from Asterias amurensis Using a Nano-encapsulation Process (나노 입자 불가사리 펩타이드의 면역 활성 증진)

  • Jeong, Hyang-Suk;Oh, Sung-Ho;Kim, Seoung-Seop;Jeong, Myoung-Hoon;Choi, Woon-Yong;Seo, Yong-Chang;Choi, Geun-Pyo;Kim, Jin-Chul;Lee, Hyeon-Yong
    • Korean Journal of Food Science and Technology
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    • v.42 no.4
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    • pp.424-430
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    • 2010
  • Immuno-modulatory activities of peptides from Asterias amurensis were investigated using a nano-encapsulation process. The molecular weights of the peptides in the range of 5-7 kDa were separated using Sephadex G-75 gel filtration. Eighty-five percent of the nano-particles were in the 300 nm range using dynamic light scattering. The cytotoxicity of the A. amurensis nano-particles against CCD-986sk human dermal fibroblast cells was 11.64% after adding 1.0 mg/mL of the samples, which was lower than that from the control (13.28% collagen). The secretion of $NO^-$ from macrophages was estimated as $40\;{\mu}M$ after adding 1.0 mg/mL of gelatin nano-particles, which was higher than the others. Prostaglandin $E_2$ production from UV-induced human skin cells decreased greatly to 860 pg/mL after adding 1.0 mg/mL of the samples. Confocal microscopy revealed that nano-particles effectively penetrated the cells within 1 hour. From these results, we consider that nano-encapsulation of the peptides from A. amurensis can improve their biological functions.

Construction of cDNA Library and EST Analysis Related to Seed-hair Characteristics in Carrot (당근 종모 형질 관련 cDNA Library 작성 및 EST 분석)

  • Oh, Gyu-Dong;Shim, Eun-Jo;Jun, Sang-Jin;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.782-789
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    • 2013
  • Carrot (Daucus carota L. var. sativa) is one of the most widely used crops in the world and is nutritionally important crop. However, seed-hair which is generated in epidermal cell of seeds causes the difficulty of the seedling process, because of the seed germination and absorption inhibitions. For these reasons, carrot seeds are commercialized after mechanical hair removal process. However, in this process, various damage and seed loss occur and breeding of hairless-seed carrot cultivar is needed to overcome these various weaknesses and additional seed production costs. In this study, cDNA libraries using 2 combinations, which were composed of short-hair seed CT-ATR 615 OP 666-13 & long-hair seed CT-ATR 615 OP 671-9, and short-hair seed CT-SMR 616 OP 659-1 & long-hair seed CT-SMR 616 OP 677-14, were constructed and EST sequences of each individuals were analyzed to reveal carrot seed-hair characteristics. Firstly, analyzed EST sequences were classified into FunCat functional categories. As a result, significant differences have been identified in metabolism category, protein folding and stabilization, protein binding, C-compound binding category from both of two combinations. Secondly, several candidate EST sequences related to seed trichome differentiation and cellulose biosynthetic process were selected based on GO data of EST sequences. These differences based on FunCat categories and candidate EST obtained by GO data analysis are thought to be involved in the formation of carrot seed hair. Finally, 741 SSR sites and 33 SNP sites were identified from analyzed EST sequences of two combinations. Then we designed SNP and SSR primer sets to develop molecular markers. These molecular markers will be used for classification of carrot cultivars and study seed-hair characteristic.

Sorghum Extract Enhances Caspase-dependent Apoptosis in Primary Prostate Cancer Cells and Immune Activity in Macrophages (수수 추출물에 의한 primary 전립선 암세포의 caspase 의존성 apoptosis 유도 및 대식세포 면역활성 증가)

  • Cho, Hyun-Dong;Kim, Jeong-Ho;Hong, Seong-Min;Lee, Ju-Hye;Lee, Yong-Seok;Kim, Du-Hyun;Seo, Kwon-Il
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1431-1437
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    • 2016
  • Sorghum bicolor L. is one of the important minor cereals in Asia, Africa, and the central United States, and it is considered a rich source of polyphenols, flavonoids, and dietary fiber. However, there is a lack of data on the anti-cancer activity of Sorghum in prostate cancer cells and immune activity in macrophages. This study aims to investigate the potential effects of an ethanol extract of S. bicolor L. (SE) on inducing apoptosis in RC-58T/h/SA#4 cells and immunomodulatory activity in RAW 264.7 cells. SE significantly inhibited the viability of RC-58T/h/SA#4 primary prostate cancer cells in a dose-dependent manner. The morphology of RC-58T/h/SA#4 cells treated with SE was shrunken and involved the formation of an apoptotic body and nuclear condensation. In addition, SE markedly activated caspase-8, -9, and -3; increased the protein levels of Bax, p53, cleaved PARP, and cytosolic cytochrome c; and decreased Bcl-2 protein expression. Furthermore, the inhibition of caspases in RC-58T/h/SA#4 cells with z-VAD-fmk attenuated SE-induced cell growth inhibition. The production of nitric oxide (NO) was also elevated by SE treatment, as revealed by immune response parameters. These results suggest that SE inhibits growth and induces apoptosis in primary human prostate cancer cells in a caspase-dependent manner, and it modulates the immune functions in macrophages. Therefore, Sorghum bicolor L. may be used as a functional food to prevent prostate cancer and enhance immune activity.

Biodegradation of Phenol by Comamonas testosteroni DWB-1-8 Isolated from the Activated Sludge of Textile Wastewater (섬유 폐수 활성 슬러지에서 분리한 Comamonas testosteroni의 생물학적 페놀 분해)

  • Kwon, Hae Jun;Choi, Doo Ho;Kim, Mi Gyeong;Kim, Dong-Hyun;Kim, Young Guk;Yoon, Hyeokjun;Kim, Jong-Guk
    • Journal of Life Science
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    • v.30 no.2
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    • pp.156-161
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    • 2020
  • Since industrialization, the production and utilization of various chemicals has contributed to improving the quality of our lives, but the subsequent discharge of massive waste is inevitable, and environmental pollution is becoming more serious every day. Exposure to chemicals as a result of environmental pollution is having a negative effect on human health and the ecosystem, and cleaning up the polluted environment that can affect our lives is a very important issue. Toxic aromatic compounds have been detected frequently in soil, groundwater, and wastewater because of the extensive use of oil products, and phenol, which is used to produce synthetic resins, textiles, and dyes, is one of the major pollutants, along with insecticides and preservatives. Phenol can cause dyspnea, headache, vomiting, mutation, and carcinogenesis. Phenol-degrading bacterium DWB-1-8 was isolated from the activated sludge of textile wastewater; this strain was identified as Comamonas testosteroni by 16S rRNA gene sequencing. The optimal culture conditions for the cell growth and degradation of phenol were 0.7% K2HPO4, 0.6% NaH2PO4, 0.1% NH4NO3, 0.015% MgSO4·7H2O, 0.001% FeSO4·7H2O, an initial pH of 7, and a temperature of 30℃. The strain was also able to grow by using other toxic compounds, such as benzene, toluene, or xylene (BTX), as the sole source of carbon.

Effects of Supplementary Copper Sources (Cu Sulfate, Cu-Methionine, Cu-Soy Proteinate) on the Performance Broiler Chickens (구리 공급원들(Cu Sulfate, Cu-Methionine, Cu-Soy Proteinate)의 첨가가 육계의 생산성에 미치는 영향)

  • Lee, Mun-Ku;Kim, Chan-Ho;Shin, Dong-Hun;Jung, Byoung-Yun;Paik, In-Kee
    • Korean Journal of Poultry Science
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    • v.38 no.2
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    • pp.121-128
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    • 2011
  • This study was conducted to investigate the effects of dietary supplementation of Cu-sulfate, Cu-methionine chelate (Cu-Met) and Cu-soy proteinate (Cu-SP) on the performance, blood parameters and mineral contents of muscle. It was conducted with a total of 1,000 one d old broilers chickens (Ross$^{(R)}$) which were assigned to four dietary treatments; Control, Cu sulfate (200 ppm Cu as $CuSO_4{\cdot}5H_2O$), Cu-Met (200 ppm Cu as Cu-methionine chelate), Cu-SP (200 ppm Cu as Cu-soy proteinate). There were significant differences (p<0.05) among treatments in weight gain. Weight gain of Cu treated groups were higher than the control during 3~5 wk. There were significant differences (p<0.05) among treatments in feed intake during 0~3 wk. Cu-Met was significantly (p<0.05) lower than the control but the differences among Cu treatments were not significant. There were significant differences (p<0.05) among treatments in feed conversion rate (FCR). Cu treated groups were lower than the control during the whole period. Production efficiency factor (PEF) was significantly higher (p<0.01) in Cu treated groups than the control. Nutrient availabilities of diets were not significantly different among the treatments. The count of white blood cell (WBC) and eosinophil (EO) were lower in Cu-SP treatment than in the control. Copper concentration in the liver was significantly (p<0.01) higher in Cu treated groups than the control. Zinc concentration in the breast and wing muscle was lower in Cu treated and that of leg muscle was higher in Cu-Met than the control. The result of this experiment showed that Cu supplementation at the level of 200 ppm as Cu sulfate, Cu-Met and Cu-SP improves weight gain (4~5 wk), FCR and PEF. Differences among Cu sources were not significant.

Production and Evaluation of Immunoreactivity of Poly Lysine-Tagged Single Chain Fragment Variable (ScFv) Lym-1 Antibody for Direct Conjugation to Fluorescence Dye (형광 물질 직접 표지를 위한 Poly Lysine 도입 Lym-1 단일사슬 항체의 제조 및 면역반응성 평가)

  • Jung, Jae-Ho;Choi, Tae-Hyun;Woo, Kwang-Sun;Chung, Wee-Sup;Kang, Joo-Hyun;Jeong, Su-Young;Choi, Chang-Woon;Lim, Sang-Moo;Cheon, Gi-Jeong
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.5
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    • pp.487-494
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    • 2009
  • Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.

Production of Citrus Plants from Ovule Cell Culture and Verification of CTV - free Plants (배주배양 세포로부터 감귤 식물체의 획득 및 감귤 트리스테자 바이러스 무병주 검증)

  • Jin, Seong Beom;Park, Jae Ho;Park, Suk Man;Lee, Dong Hoon;Yun, Su Hyun
    • Horticultural Science & Technology
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    • v.35 no.1
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    • pp.121-130
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    • 2017
  • This study was carried out to investigate a method for producing cultured virus - free ovules for breeding high - quality Citrus cultivars. Ovules from the immature fruits of three citrus cultivars native to Jeju (Dongjeongkyool, Cheongkyool, and Jikak) and two cultivars of Citrus unshiu Marc. (Miyagawa wase and Haryejosaeng) that were thought to be infected with Citrus tristeza virus (CTV) were cultured on MS2 medium (Murashige - Skoog [MS] basal medium containing $500mg{\cdot}L^{-1}$ malt extract, $50g{\cdot}L^{-1}$ sucrose, $1.0 mg{\cdot}L^{-1}$ kinetin, and $8g{\cdot}L^{-1}$ agar). After four weeks of culture, 10, 21, 13, 5, and 7 somatic embryos and 2, 4, 2, 4, and 5 white callus cells (surrounding green somatic embryos) were obtained from Dongjeongkyool, Cheongkyool, Jikak, Miyagawa wase, and Haryejosaeng, respectively. After six weeks of culture, somatic embryos were obtained from cultured cells grown on MT basal medium supplemented with malt extract ($500mg{\cdot}L^{-1}$), lactose ($70g{\cdot}L^{-1}$), and agar ($16g{\cdot}L^{-1}$). Over 60% of the somatic embryos from citrus cultivars native to Jeju developed into normal plants on MS basal medium supplemented with malt extract ($500mg{\cdot}L^{-1}$), sucrose ($50g{\cdot}L^{-1}$), and agar ($8g{\cdot}L^{-1}$) after 10 weeks of culture. Normal plants were regenerated from two Citrus unshiu Marc. cultivars on MT basal medium supplemented with sorbitol (1.0 M), galactose (1.0 M), $GA_3$ ($1.0mg{\cdot}L^{-1}$), and Gelrite ($3g{\cdot}L^{-1}$). The absence of virus in plants generated from cultured ovules was confirmed by RT - PCR and antigen - antibody reactions. Therefore, virus - free Citrus cells can be obtained for breeding high - quality citrus cultivars using the biotechnological technique evaluated in this study.

Physiological Activity and Nutritional Composition of Pleurotus Species (느타리속 버섯류의 영양성분 및 생리활성)

  • Um, Su-Na;Jin, Gyoung-Ean;Park, Kye-Won;Yu, Young-Bok;Park, Ki-Moon
    • Korean Journal of Food Science and Technology
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    • v.42 no.1
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    • pp.90-96
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    • 2010
  • In this study, the anti-oxidant, anti-tumorigenic, anti-hypertensive, anti-thrombic, anti-diabetic, and anti-inflammatory properties of 18 different species of genus Pleurotus were investigated. In addition, the amino acid, $\beta$-glucan, and polyphenol content were also measured. All species contained more than 20 mg% of polyphenol with the highest contents found in Pleurotus cornucopiae var. citrinopileatus (yellow pleurotus) ($39.13{\pm}0.82\;mg%$). The $\beta$-glucan contents was also the highest in yellow Pleurotus ($37.67{\pm}0.22%$) followed by Won-Hyeong1 (C, $28.75{\pm}0.61%$) and Jang-an PK (A, $27.95{\pm}0.33%$). The yellow Pleurotus exhibited the highest antioxidant activity as assessed by the DPPH scavenging rate with an $IC_{50}$ value of $2.94{\pm}0.44\;mg/mL$. Ethanol extracts from the yellow Pleurotus treated at 1% concentration showed cytotoxic activity up to 36.9% in the human embryonic kidney 293T cell lines. The yellow Pleurotus also showed the highest inhibitory effects on ACE activity ($60.52{\pm}0.2%$). Finally, the yellow Pleurotus exhibited anti-diabetic and anti-inflammatory properties as shown by inhibition of $\alpha$-amyloglucosidase activity ($50.5{\pm}0.8%$) and nitric oxide production ($68.4{\pm}0.3%$). Taken together, our data indicate the yellow pleurotus is a promising functional food ingredients.

Discontinuous Percoll Gradients Enrich X-Bearing Porcine Sperms and Female Embryos (불연속 Percoll 원심분리에 의한 돼지 X-정자와 자성배아에 관한 연구)

  • Shim, Dae-Yong;Yoo, Seong-Jin;Kang, Han-Seung;Yoo, Jeong-Min;Lee, Chae-Kwan;Kang, Sung-Goo
    • Development and Reproduction
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    • v.5 no.1
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    • pp.47-52
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    • 2001
  • Predetermination of sex in livestock of offpring is in great demand and is of critical importance to providing for the most efficient production of the animal ariculture. Such a sexing techlology would also enhance the economy of conventional artificial insemination(AI) and aid the porcine industry. The purpose of this study was to evaluate the efficiency of enriching X-bearing porcine sperm using discontinuous percoll gradients and PCR mefhod. Semen was collected from mature boars of proven fertility center (AI center KimHae). Sperm was leaded on the isotonic discontinuous percoll gradient and then it was centrifuged at 120 ${\times}$ g for 20 minutes. After centrifugation, sperm included in each fraction were recovered (7${\times}$10$^6$ sperms/ml) and then sperm genomic DNA was extractedfor the PCR. SRY gene was used to evaluate the ratio between X and Y sperm in the separated fractions. Ju viro ffrtilization wascarried out by adding the unseparated sperm (control) or separated (experimental poop) to the matured oocytes in TCM-199. Embryos for sex determination were obtained at 2 cell stage and then was used for SRY gene amplification. After centrifugation of discontinuous percoll gradient, the most motile sperm was obtained at 95% fiaction (94.4% ${\pm}$ 5.1%, p < 0.01). The PCR analysis evaluated that 30%, 50% and 65% fractions were Y sperm rich, whereas 80% and 95% fractions were X sperm rich. PCR analysis with each porcineembryo showed that 33.3% of control and 66.7% of experimental group were determined as female embryos. In conclusion, in vitro matured oocytes inseminated with sperms (95% fraction) prepared by percoll gradient centrifugation showed high fertilization rates and female embryos than control sperms.

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