• Journal of Chest Surgery
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• 제42권2호
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• pp.157-164
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• 2009

배경: 폐동맥 판막 협착 혹은 형성 부전을 동반한 여러 선천성 심장기형의 수술적 치료를 위하여, 다양한 종류의 우심실-폐동맥간 도관이 사용되었다. 장기 성적이 우수한 냉동동종 이식편(cryopreserved homograft)의 공급의 제한으로 이를 대체할 이종 조직 이식편의 석회화 방지를 위한 효과적인 기법의 확립이 필요하다. 본 연구에서는 L-lysine을 이용한 diamine bridge의 항석회화 효과를 Ethanol과 비교하여 알아보고자 하였다. 대상 및 방법: 0.625% glutaraldehyde ($4^{\circ}C$에서 2일, 상온에서 7일간) 고정한 돼지 심낭을 80% Ethanol (상온에서 1일), 혹은 0.1M L-lysine ($37^{\circ}C$에서 2일)로 처리 한 후 각각의 두께(thickness)와 장력(tensile strength)을 측정하였다. 각각의 항석회화 처리한 돼지 심낭을 생후 3주된 쥐의 피하조직에 이식하고 8주 뒤 칼슘을 정량하고 조직학적 소견을 관찰하였다. 결과: 0.625% glutaraldehyde 고정만 시행한 군$(51.2{\pm}8.5ug/mg)$과 비교하여 0.625% glutaraldehyde 고정 후에 80% Ethanol 처리한 군($13.6{\pm}10.0ug/mg$, p=0.008)과, 0.625% glutaraldehyde 고정 후에 L-lysine 처리한 군($15.3{\pm}1.0ug/mg$, p=0.002), 그리고 0.625% glutaraldehyde 고정 후에 80% Ethanol과 L-lysine 처리한 군($16.1{\pm}11.1ug/mg$, p=0.012)은 통계적으로 의미 있게 칼슘의 침착량이 적었다. 0.625% glutaraldehyde 고정 후에 80% Ethanol 처리한 군의 두께와 장력은 각각 $0.18{\pm}0.02mm,\;1.20{\pm}0.30kg$중/5mm로 0.625% glutaraldehyde 고정 후에 L-lysine 처리한 군의 $0.13{\pm}0.03mm$, $0.85{\pm}0.36$ 1.0kg 중/5mm 보다 증가되어 있었다(p<0.01, p=0.035). 결론: L-lysine을 이용한 diamine bridge는 Ethanol과 비교하여 비슷한 항석회화 효과를 보여 주었으며 Cross-link를 증가시켜 이종 이식편의 두께와 장력을 증가시켜 주는 효과가 있었다.

• 한국염색가공학회지
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• 제30권4호
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• pp.264-274
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• 2018

Immobilization of lysozyme on chitosan non-woven using glutaraldehyde(GA) was investigated. For this, 100 % chitosan non-woven was prepared as novel support for the enzyme immobilization. In addition, free lysozyme activity was examined depending on various pH and temperature by measuring time. Moreover, the optimum immobilization conditions depending on various pH, temperature, immobilization time and lysozyme concentration was evaluated. In addition, thermal stability and storage stability of immobilized lysozyme were measured. The characteristics of immobilized lysozyme was examined by FT-IR, surface morphology, and MTT assay. The results are follows: the optimal immobilization of lysozyme were pH 7.0, $25^{\circ}C$, lysozyme concentration 1.5 mg/ml, immobilization time 240 min. The immobilized lysozyme showed higher thermal stability than the free trypsin. The immobilized lysozyme activity was retained 80 % of its initial activity at $4^{\circ}C$ over 30 days of storage. The lysozyme was immobilized effectively on chitosan non-woven by observation of surface morphology.

• Journal of Microbiology and Biotechnology
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• 제22권11호
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• pp.1486-1493
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• 2012

In this study, we investigated whether galactooligosaccharide (GOS) can be stably and steadily synthesized using immobilized ${\beta}$-galactosidase (${\beta}$-gal) inclusion body (IB)-containing E. coli cells during long-term repeated-batch operation. To improve the operational stability of this enzyme reactor system, immobilized E. coli cells were crosslinked with glutaraldehyde (GA) after immobilization of the E. coli. When we treated with 2% GA for E. coli crosslinking, GOS production continued to an elapsed time of 576 h, in which seven batch runs were operated consecutively. GOS production ranged from 51.6 to 78.5 g/l ($71.2{\pm}10.5$ g/l, n = 7) during those batch operations. In contrast, when we crosslinked E. coli with 4% GA, GOS production ranged from 31.5 to 64.0 g/l ($52.3{\pm}10.8$, n = 4), and only four consecutive batch runs were operated. Although we did not use an industrial ${\beta}$-gal for GOS production, in which a thermophile is used routinely, this represents the longest operation time for GOS production using E. coli ${\beta}$-gal. Improved stability and durability of the cell immobilization system were achieved using the crosslinking protocol. This strategy could be directly applied to other microbial enzyme reactor systems using cell immobilization to extend the operation time and/or improve the reactor system stability.

• Journal of Chest Surgery
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• 제44권2호
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• pp.99-107
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• 2011

Background: Calcification is the most frequent cause of clinical failure of bioprosthetic tissues fabricated from GA-fixed porcine valves or bovine pericardium. A multi-factorial approach using different mechanisms was recently developed to reduce the calcification of bioprosthetic tissues. The purpose of the present study was to evaluate the synchronized synergism of using L-arginine and $NaBH_4$, compared with ethanol and L-lysine, in glutaraldehyde treated porcine pericardium from the standpoint of calcification and tissue elasticity. Materials and Methods: Porcine pericardium was fixed at 0.625% GA (7 days at room temperature after 2 days at $4^{\circ}C$). An interim step of ethanol (80%; 1 day at room temperature) or L-lysine (0.1 M; 2 days at $37^{\circ}C$) or L-arginine (0.1 M; 2 days at $37^{\circ}C$) was followed by completion of the GA fixation. A final step of NaBH4 (0.1 M; 2 days at room temperature) was followed. Their tensile strength, thickness, and thermal stability were measured. Treated pericardia were implanted subcutaneously into three-week-old Sprague-Dawley rats for 8 weeks. Calcium content was assessed by atomic absorption spectroscopy and histology. Results: L-arginine and $NaBH_4$ pretreatment ($1.81{\pm}0.39$ kgf/5 mm p=0.001, $0.30{\pm}0.08$ mm p<0.001) significantly increased tensile strength and thickness compared with the control ($0.53{\pm}0.34$ kgf/5 mm, $0.10{\pm}0.02$ mm). In a thermal stability test, L-arginine and $NaBH_4$ pretreatment ($84.25{\pm}1.12^{\circ}C$, p=0.023) caused a significant difference from the control ($86.25{\pm}0.00^{\circ}C$). L-lysine and $NaBH_4$ pretreatment ($183.8{\pm}42.6$ ug/mg, p=0.804), and L-arginine and $NaBH_4$ pretreatment ($163.3{\pm}27.5$ ug/mg, p=0.621) did not significantly inhibit calcification compared to the control ($175.5{\pm}45.3$ ug/mg), but ethanol and $NaBH_4$ pretreatment did ($38.5{\pm}37.3$ ug/mg, p=0.003). Conclusion: The combined pretreatment using L-arginine and $NaBH_4$ after GA fixation seemed to increase the tensile strength and thickness of porcine pericardium, fixed with GA. Additionally, it seemed to keep thermal stability. However it could not decrease the calcification of porcine pericardium fixed with GA. $NaBH_4$ pretreatment seemed to decrease the calcification of porcine pericardium fixed with GA, but only with ethanol.

• 폴리머
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• 제38권6호
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• pp.687-693
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• 2014

분자량이 400 g/mol인 poly(ethylene glycol)(PEG 400)을 poly(ether-block-amide) 1657(PEBAX 1657)와 혼합을 통하여 막을 제조하였고, time-lag 법에 의해 $N_2$, $O_2$, $CH_4$, $CO_2$, 그리고 $SO_2$ 기체에 대한 투과도를 테스트하였다. 투과특성은 막 내에서 기체분자이동에 어느 것이 지배적인가를 알기 위하여 확산과 용해 항으로 조사하였다. PEG 400의 함량이 증가함에 따라 모든 기체에 대한 투과도 및 이상선택도가 모두 증가하였다. 특히 $CO_2/N_2$의 경우 53.2(PEG 400이 첨가되지 않은 PEBAX 1657)으로부터 84.1(PEG 400이 50% 첨가된 막)이었으며, $SO_2/CO_2$는 38.9 에서 50.7, 그리고 $CO_2/CH_4$의 경우는 177.7에서 31.4의 결과를 보여주었다. 투과도와 선택도의 증가는 기체들의 용해도로 인한 것이며, 특히 $CO_2$와 $SO_2$에 대해서는 더욱 증가한다는 것을 알 수 있었다. 수증기에 대한 내구성을 얻기 위하여, glutaraldehyde(GA)가 PEBAX 1657/PEG 400 혼합막에 첨가되었다. 결과적으로 투과도는 혼합막에 있어서 투과도 증가와 선택도 감소의 주된 요인으로 작용하는 자유부피와 ether oxide 기의 감소로 인하여 줄어들었으나 이는 막의 내구성을 향상시키는데 도움이 되었을 것으로 사료된다.

• Journal of Chest Surgery
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• 제32권11호
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• pp.998-1003
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• 1999

Background: Bovine pericardium treated with glutaraldehyde(GA) is one of the most popular prosthetic materials. However, its late calcific degeneration after implantation results in early failure of the prosthesis. Therefore, we investigated the effects of combined treatment with sodium dodecyl sulfate(SDS) and glutamate on calcific degeneration of GA treated bovine pericardium. Material and Method: Sixty square-shaped pieces of bovine pericardia were fixed in 0.625% GA solution with 4g/L MgCl2.6H2O as a control group (group 1). Sixty pieces pretreated with 1% SDS (group 2) and sixty pieces posttreated with 8% glutamate (group 3) were also fixed in the same GA solution. Sixty pieces pretreated with 1% SDS and posttrated with 8% glutamate were also fixed in the same GA solution (group 4). After 1 month of fixation, the pieces were implanted into the belly of sixty Sprague-Dawley rat subdermally and were extracted 1 month, 2 months and 3 months after the implantation. With an atomic absorption spectrophotometry, we measured the calcium amount deposited. Result: The calcium deposition in 1 month was 2.01$\pm$0.13 mg/g in group 1, 1.45$\pm$0.31 mg/g in group 2, 2.49$\pm$0.15 mg/g in group 3 and 0.75$\pm$0.27 mg/g in group 4. In 2 months, it was 3.57$\pm$0.15 mg/g in group 1, 0.98$\pm$0.30 mg/g in group 2, 3.46$\pm$0.12 mg/g in group 3, and 1.48$\pm$0.39 mg/g in group 4, and 5.45$\pm$0.42 mg/g in group 1, 2.43$\pm$0.53 mg/g in group 2, 4.20$\pm$0.55 mg/g in group 3, and 1.02$\pm$0.27 mg/g in group 4 in 3 months. The calcium depositions in group 2 and 4 were less than those of group 1 and 3 in 1 month 2, months, and 3 months(p<0.01). The calcium depositions in group 1, 2 and 3 increased with time. However, they remained unchanged in group 4, which was statistically significant(p<0.01). Conclusion: Pretreatment with SDS is effective in reducing calcification of GA treated bovine pericardium, and the combined method of pretreatment with SDS and posttreatment with glutamate was more effective than the other methods.

• Journal of Chest Surgery
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• 제29권1호
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• pp.7-13
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• 1996

The conventional glutaraldehyde (GA) fixation method of tissue valves is considered to be responsible for accelerated valve degeneration. The release of toxic GA from the valve tissue is believed to limit endothelial cell (EC) ingrowth. Removal of toxic GA by reaction with L-glutamic acid and storage in a Paraben solution may offer good EC growth. To investigate the conditions for endothelialization of tissue valves, the growth properties of ECs on the conventionally and alternatively treated pericardial tissue were compared. Conventional preparation included zero-pressure fixation for 72 hours in phosphated-buffered saline (PBS) solution containing 0.5% GA at 4$^{\circ}C$ and storage into PBS containing 0.2% GA(group I). Alternatively treated pericardial tissues were divided into three postfixation treatment groups : (1) storage in PBS solution containing Paraben(group II), (2) treatment with PBS containing 8$^{\circ}C$ L-glutamic acid(PH 7.35) and storage in PBS solution containing Paraben (g oup III), (3) treatment with L-glutamic acid dissolved in distilled water (PH 3.5) (group IV). Pericardial tissue were transferred into the 24-well plate after storage for 4 weeks. ECs were harvested enzymatically from the bovine pulmonary artery and grown to confluence on culture flask surfaces. Detached ECs by trypsin were incubated into the each well of the 24-well plate including test pericardial tissues. Cells were detached by trypsin, 1, 2, 3, 5, 7 days after incubation and counted on the hemacytometer. Cell viability test was performed by frypan-blue exclusion method. Acute cell death in the group I were found even after prolonged washing. The group II showed prolonged cell survival compared with the group I. Both group III and group IV showed better cell growth than group II. There was no statistically significant difference between group III and group IV method in terms of EC growth. This results suggest that treatment by L-glutamic ac id and storage in a Paraben solution be a promising approach for improvement of durability of GA-treated tissue valves.

• Journal of Chest Surgery
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• 제36권3호
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• pp.131-135
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• 2003

Glutaraldehyde (GA)에 보존한 이종보철편들은 생체 이식 후 석회화 변성을 보이는 문제점을 나타내므로 이 단점을 보완하기 위해 연구자는 GA 용액에 Mg염을 첨가 칼슘의 반응을 막아 석회화를 완화하며 더불어 chitosan 및 glutamate 등의 아미노산으로 후처치하여 석회화 완화를 극대화하고자 하였다. 대상 및 방법: 우심낭을 정방형 조각으로 만들어 40조각의 우심낭편은 대조군으로 MgC1$_2$6$H_2O$를 4g/L 첨가한 0.625% GA 용액에 보존하고(1군), 40조각은 이 GA 용액에 보관하다 이식 24시간 전 2% chitosan 용액에 옮겨 보존(2군), 또 다른 40조각 역시 같은 방법으로 8% glutamate 용액에 후처치(3군)하여, 이들을 40마리의 백서 복부 피하에 각각 한 조각씩 이식하여 1개월, 2개월, 3개월 및 4개월째에 적출, spectophotometry로 침착한 칼슘을 정량하였다. 결과： 이식 후 1개월 및 2개월에는 세균간의 석회화 정도에 차이가 없었으나. 3개월째에는 1군 5.45$\pm$0.42 mg/g, 2군 3.22$\pm$1.31 mg/g, 3군 4.20$\pm$0.55 mg/g의 통계적 차이를 보여, 2군에서 유의한 석회화 완화 효과가 관찰되고 있고 (p<0.05), 4개월째에는 1군 6.01$\pm$1.21 mg/g, 2군 3.78$\pm$1.82 mg/g, 3군 3.92$\pm$0.92 mg/g으로 2, 3군에서 유의한 석회화 완화 효과를 보여주고 있다(p<0.05). 결론: 즉 2% chitosan 후처치로 3개월의 시간이 경과함에 따라 피하에 이식된 보철편의 석회화가 완화되었으며 8% glutamate는 4개월에서 그 완화효과를 보여주고 있다.다. 현대의 실정 에 맞는 과학적 제조방법을 이용하여 산업화한다면 재래식으로 인해 걸리는 시간, 노력 에 걸맞은 가격, 품질 등을 유지해 나갈 수 있으리라 본다.는 음식 10위를 조사한 결과 선호음식의 경우 저체중군과 정상군의 경우 순위는 달랐지만 10위에 선별된 음식들이 주로 과 (수박, 귤, 딸기, 복숭아, 바나나, 포도, 사과)과 음료수(사이다, 환타) 등 열량밀도가 낮고 수분함량이 높은 식품인 반면 과체중군은 햄버거, 돈까스, 튀김 만두, 샌드위치, 쇠고기 및 돼지고기 구이 등이 선호음식 10위내에 포함되어 정상군이나 저체중군보다 고지방.고단백 음식들의 기호도가 과체중군에서 높음을 확인하였다., 단맛을 좋아하는 사람이 싫어하는 사람보다 열량과 당질섭취량은 유의 적으로 적게, 지방 섭취량은 유의적으로 높았다. 짠맛을 좋아하는 사람은 나트륨 섭취량이 7890 mg으로 짠맛을 싫어하는 사람 5579 mg보다 유의적 (p < 0.05)으로 많은 양을 섭취하였다.고, Ca : P 중량비가 각각 2.63$\pm$1.99, 1.79$\pm$0.39, 2.80$\pm$0.53, 8.35$\pm$12.87이었으며, Ca : Mg 중량비는 각각 11.86$\pm$540, 9.29$\pm$1.34, 9.09$\pm$2.09, 32.50$\pm$41.35이었다. 칼슘과 마그네슘의 흡수에 영향을 미치는 P : Mg 분자량비는 곡류가공품이 4.11$\pm$1.54, 라면류가 4.17$\pm$0.67, 레토르트식품은 2.58$\pm$0.45, 음료류는 2.59$\pm$2.50으로 나타났다. 위의 연구결과에서 볼 때 시료의 93.0%가 실제측정값이 표시량의 80% 이상이어야

• Macromolecular Research
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• 제11권4호
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• pp.291-295
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• 2003

Dextran as a candidate material for colon-specific drug delivery has been studied. Crosslinked dextran hydrogels were prepared by mixing dextran, $MgCl_2$, glutaraldehyde (GA) and polyethyleneglycol (PEG 400) in water. The dextran hydrogels were characterized by measuring equilibrium swelling ratios and mechanical strengths. Response surface methodology (Central Composite Design) was used to evaluate the swelling behaviors and mechanical strengths as functions of concentrations of $MgCl_2$, GA, and PEG 400, which was found to be useful for the evaluation. It showed that the swelling behavior and mechanical strengths were influenced significantly by PEG 400 and $MgCl_2$ concentrations.

• Journal of Chest Surgery
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• 제47권4호
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• pp.333-343
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• 2014

Background: A preclinical study was conducted for evaluating a valved conduit manufactured with a glutaraldehyde (GA)-fixed bovine pericardium treated using an anticalcification protocol. Methods: Bovine pericardia were decellularized, fixed with GA in an organic solvent, and detoxified. We prepared a valved conduit using these bovine pericardia and a specially designed mold. The valved conduit was placed under in vitro circulation by using a mock circulation model, and the durability under mechanical stress was evaluated for 2 months. The valved conduit was implanted into the right ventricular outflow tract of a goat, and the hemodynamic, radiologic, histopathologic, and biochemical results were obtained for 6 months after the implantation. Results: The in vitro mock circulation demonstrated that valve motion was good and that the valved conduit had good gross and microscopic findings. The evaluation of echocardiography and cardiac catheterization demonstrated the good hemodynamic status and function of the pulmonary xenograft valve 6 months after the implantation. According to specimen radiography and a histopathologic examination, the durability of the xenografts was well preserved without calcification at 6 months after the implantation. The calcium and inorganic phosphorus concentrations of the explanted xenografts were low at 6 months after the implantation. Conclusion: This study demonstrated that our synergistic employment of multiple anticalcification therapies has promising safety and efficacy in the future clinical study.