• Reproductive and Developmental Biology
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• 제31권2호
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• pp.103-108
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• 2007

This study was carried out to investigate the effects of cryoprotectants, warming solution and removal of lipid on open pulled straw vitrification (OPS) method of porcine embryos produced by nuclear transfer (NT) of fetal fibroblasts. All solutions used during vitrification were prepared with holding medium consisting of 25 mM Hepes buffered TCM199 medium containing 20% fetal bovine serum (FBS) at $38.5^{\circ}C$. The blastocysts derived from NT with or without lipid were vitrified in each medium of different concentrations of dimethyl sulfoxide (DMSO) and ethylene glycol (EG). Also, blastocysts after cryopreservation were warmed into different concentrations of sucrose in warming solution. The optimal concentrations of cryoprotectants in vitrification solution were 10% DMSO + 10% EG in vitrification solution 1 (VS1) and 20% DMSO + 20% EG in vitrification solution 2 (VS2). The optimal concentrations of sucrose were 0.3 M sucrose in warming solution 1 (WS1) and 0.15 M sucrose in warming solution 2 (WS2). lipid removal from oocytes before NT enhanced the viability of NT embryos after vitrification. Our results show that use of the OPS method in conjunction with lipid removal provides effective cryopreservation of porcine nuclear transfer embryos.

• 분석과학
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• 제18권6호
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• pp.559-567
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• 2005

최근 관심이 되고 있는 혈액 중 다이옥신 분석방법을 마련하기 위해 미국 NIST의 혈청 인증표준물질 1589a를 이용하여, 캐나다, 일본 등 선진국의 혈액 중 다이옥신 분석방법을 비교 검토하고, 우리나라 실정에 적합한 혈액 중 다이옥신 분석방법(안)을 제시하였다. 제안된 분석방법은 추출과정에서 유분 등 지방의 효과적인 제거를 위한 알칼리 분해 추출 후 기존의 다이옥신 전처리방법과 유사한 방법으로 정제하여 두 종류의 컬럼을 사용하는 분석방법이다. 또한, 혈청 인증표준물질과 실제 혈액시료에 제안된 분석방법을 적용한 결과 정량한계는 사염화물(TeCDD/DF) 및 오염화물(PeCDD/DF)이 1 pg/g-lipid, 육염화물(HxDD/DF) 및 칠염화물(HpCDD/DF)이 2 pg/g-lipid 그리고 팔염화물(OCDD/DF)이 4 pg/g-lipid이며, 검량용 표준물질은 검출 농도를 고려하여 $CS_L$(0.1~1), $CS_1/2$(0.25~2.5), $CS_1$(0.5~5.0), $CS_2$(2~20) 및 $CS_3$(10~100)로 제시하였다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권4호
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• pp.315-321
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• 2005

Supercritical carbon dioxide extraction was investigated as a method for removing lipids and bad flavor from tuna viscera. To find the optimum conditions, different experimental variables, such as pressure, temperature, flow rate of solvent and sample size, were evaluated for the effective removal of lipids and the undesirable smell. Ethanol was used as the entrainer, with a $3\%$ by vol $CO_2$ flow rate. By increasing the pressure at constant temperature, the efficiency of the lipid removal was improved and the protein was concentrated without denaturalization. The main fatty acids extracted from the tuna viscera were palmitic acid (16:0), heptadecanoic acid (17:1), oleic acid (18:1) and docosahexaenoic acid (22:6). The major amino acids in the tuna viscera treated by supercritical carbon dioxide were glutamic acid, leucine and lysine, and the free amino acids were L-proline, taurine and L-$\alpha$-aminoadipic acid.

• 한국축산식품학회지
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• 제37권1호
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• pp.29-37
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• 2017

Defatted bovine liver (DBL) is a potential source of protein and minerals. Supercritical carbon dioxide ($SC-CO_2$) and a traditional organic solvent method were used to remove lipid from bovine liver, and the quality characteristics of a control bovine liver (CBL), bovine liver defatted by $SC-CO_2$ ($DBLSC-CO_2$) at different pressures, and bovine liver defatted by organic solvent (DBL-OS) were compared. The $DBLSC-CO_2$ samples had significantly higher (p<0.05) protein, amino acid, carbohydrate, and fiber contents than CBL and DBL-OS. There was a higher yield of lipid from CBL when using $SC-CO_2$ than the organic solvent method. SDS-PAGE analysis demonstrated that the CBL and $DBLSC-CO_2$ had protein bands of a similar intensity and area, whereas DBL-OS appeared extremely poor bands or no bands due to the degradation of proteins, particularly in the 50 to 75 kDa and 20 to 25 kDa molecular weight ranges. In addition, $DBLSC-CO_2$ was shown to have superior functional properties in terms of total soluble content, water and oil absorption, and foaming and emulsification properties. Therefore, $SC-CO_2$ treatment offers a nutritionally and environmentally friendly approach for the removal of lipid from high protein food sources. In addition, $SC-CO_2$ may be a better substitute of traditional organic solvent extraction for producing more stable and high quality foods with high-protein, fat-free, and low calorie contents.

• 대한임상독성학회지
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• 제1권1호
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• pp.6-11
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• 2003

Various forms of dialytic techniques are available for detoxification. Hemodialysis, hemoperfusion and hemofiltration (hemodialfiltration) are the main treatment modalities. Because these modalities are rather invasive and expensive, it must be decided in balance of the risk and benefit to the patient. The prime consideration in the decision is based on the clinical features of poisoning; hemodialysis or hemoperfusion should be considered in general if the patient's condition progressively deteriorates despite intensive supportive therapy. The hemodialysis technique relies on passage of the toxic agent through a semipermeable membrane so that it can equilibrate with the dialysate and subsequently removed. It needs a blood pump to pass blood next to a dialysis membrane, which allows agents permeable to the membrane to pass through and reach equilibrium. Solute (or drug) removal by dialysis has numerous determinants such as solute size, its lipid solubility, the degree to which it is protein bound, its volume of distribution etc. The technique of hemoperfusion is similar to hemodialysis except there is no dialysis membrane or dialysate involved in the procedure. The patient's blood is pumped through a perfusion cartridge, where it is in direct contact with adsorptive material (usually activated charcoal) that has a coating material such as cellulose. This method can be used successfully with lipid-soluble compounds and with higher-molecular-weight compounds than for hemodialysis. Protein binding does not significantly interfere with removal by hemoperfusion. In conclusion, hemodialysis, hemoperfusion and hemofiltration can be used effectively as adjuncts to the management of severely intoxicated patients.

• 한국축산식품학회지
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• 제35권4호
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• pp.466-472
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• 2015

A rapid and simple analytical method, using liquid chromatography tandem mass spectrometry (LC-MS/MS), was developed to detect myo-inositol (MI) in infant formulas. For protein removal: acid hydrolysis and lipid removal through organic solvent extraction. The operating conditions for instrumental analysis were determined based on previously reported analogous methods that used LC-MS/MS. Quantitative analysis was used for the detection limit test, infant formula recovery test, and standard reference material (SRM) 1849a to verify the validity of our LC-MS/MS analytical method, which was developed to quantify MI. For validation, the results of our method were compared with the results of quantitative analyses of certified values. The test results showed that the limit of detection was 0.05 mg/L, the limit of quantitation was 0.17 mg/L, and the method detection limit was 17 mg/kg. The recovery test exhibited a recovery between 98.07-98.43% and a relative standard deviation between 1.93-2.74%. Therefore, the result values were good. Additionally, SRM 1849a was measured to have an MI content of 401.84 mg/kg and recovery of 98.25%, which is comparable to the median certified value of 409 mg/kg. From the aforementioned results, we judged that the instrumental analysis conditions and preparation method used in this study were valid. The rapid analytical method developed herein could be implemented in many laboratories that seek to save time and labor.

• Annals of Laboratory Medicine
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• 제38권6호
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• pp.518-523
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• 2018

Background: Lipemia, a significant source of analytical errors in clinical laboratory settings, should be removed prior to measuring biochemical parameters. We investigated whether lipemia in serum/plasma samples can be removed using a method that is easier and more practicable than ultracentrifugation, the current reference method. Methods: Seven hospital laboratories in Spain participated in this study. We first compared the effectiveness of ultracentrifugation ($108,200{\times}g$) and high-speed centrifugation ($10,000{\times}g$ for 15 minutes) in removing lipemia. Second, we compared high-speed centrifugation with two liquid-liquid extraction methods-LipoClear (StatSpin, Norwood, USA), and 1,1,2-trichlorotrifluoroethane (Merck, Darmstadt, Germany). We assessed 14 biochemical parameters: serum/plasma concentrations of sodium ion, potassium ion, chloride ion, glucose, total protein, albumin, creatinine, urea, alkaline phosphatase, gamma-glutamyl transferase, alanine aminotransferase, aspartate-aminotransferase, calcium, and bilirubin. We analyzed whether the differences between lipemia removal methods exceeded the limit for clinically significant interference (LCSI). Results: When ultracentrifugation and high-speed centrifugation were compared, no parameter had a difference that exceeded the LCSI. When high-speed centrifugation was compared with the two liquid-liquid extraction methods, we found differences exceeding the LCSI in protein, calcium, and aspartate aminotransferase in the comparison with 1,1,2-trichlorotrifluoroethane, and in protein, albumin, and calcium in the comparison with LipoClear. Differences in other parameters did not exceed the LCSI. Conclusions: High-speed centrifugation ($10,000{\times}g$ for 15 minutes) can be used instead of ultracentrifugation to remove lipemia in serum/plasma samples. LipoClear and 1,1,2-trichlorotrifluoroethane are unsuitable as they interfere with the measurement of certain parameters.

• KSBB Journal
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• 제15권2호
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• pp.150-155
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• 2000

본 연구는 Streptomyces sp. BH-405의 배양액으로부터 유기용매로 추출하여 지질과 산화에 대한 항산화 효과를 실험하였다. Streptomyces sp. BH-405의 배양액을 ethylether로 추출한 후 silica gel column(100$\times$10 cm) chromatography로 분획한 획분을 POV로 측정한 결과 fraction 3에서 향산화 활성이 가장 높았다. 항산화 활성이 높은 fraction 3 획분을 다시 preparative TLC와 column chromatography로 더욱 정제하여 band 2를 얻었다. Band 2의 항산화 효과를 알아보기 위해 기존의 항산화제와 비교한 결과 이-$\alpha$-tocopherol, BHA보다 산화억제 효과가 높았으며 DPPH에 의한 수소 공여능은 실험결과가 우수하였다. 한편 band 2는 쥐 간의 microsome에 있어서 유리기의 소거기능이 있었고 NADPH 및 Fenton 시액에 위한 효소적 과산화 지질의 생성을 억제하였다. 그리고 mitochondria와 리놀산의 과산화 유도계에서도 지질의 과산화 과정에 생성하는 유리기의 형성을 차단하였다. 결론적으로 Streptomyces sp. BH-405의 배양액에서 얻은 획분인 band 2는 산화를 억제하는 활성이 있었다.

• 한국식품위생안전성학회지
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• 제33권5호
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• pp.354-360
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• 2018

견과류 중에 있는 트리코테센계 곰팡이독소 오염도를 조사하기 위하여 LC-MS/MS를 이용한 정확성과 신뢰성을 동시에 확보할 수 있는 분석방법을 개발하였다. 견과류 중 트리코테센계 곰팡이 독소는 QuEChERS 추출 및 EMR-Lipid-dSPE 정제과정을 통하여 분석에 사용되었다. 검량선 작성을 위하여 트리코테센계 곰팡이독소 10종에 대하여 $2.00{\sim}75.00{\mu}g/kg$의 범위로 혼합표준용액을 제조하여 실험하였으며, 상관계수는 모두 0.998 이상으로 높은 직선성을 나타내었다. 분석방법의 검출한계는 $0.41{\sim}3.57{\mu}g/kg$로 나타났으며, 정량한계는 $1.23{\sim}10.82{\mu}g/kg$로 나타났다. 또한 트리코테센계 곰팡이 독소 10종에 대하여 각각 저, 중, 고 3가지 농도로 처리하여 회수율 실험을 수행한 결과 81.84~96.87%로 나타났다. 확립된 분석법으로 견과류 중 땅콩을 대상으로 오염도를 조사한 결과 1종에서 deoxynivalenol이 검출되었다. 이러한 결과를 바탕으로 확립된 시험법은 견과류 중 트리코테센계 곰팡이 독소 분석에 적합함을 확인할 수 있었으며 견과류 중 트리코테센계 곰팡이 독소 검출 가능성을 확인한바 보다 다양한 종류의 견과류에 대한 모니터링 조사가 필요한 것으로 판단된다.

• Journal of Plant Biotechnology
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• 제42권2호
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• pp.123-128
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• 2015

Pregermination treatment of cocoa beans either with the testa, group PCB (+T), or without the testa, group PCB (-T), was studied here to determine whether this treatment (incubation up to 120 h at $25^{\circ}C$) has any effect on the levels of allergenic proteins or on chemical composition. Our proximate analysis included carbohydrates, proteins, and lipids. We used western blotting to measure changes in the amounts of allergenic proteins in the cocoa beans during the pregermination treatment. The protein and carbohydrate content of both groups (with or without the testa) decreased slightly during this treatment, whereas lipid content increased. Group PCB (-T) showed more rapid metabolic processes than did group PCB (+T) during the pregermination treatment. Western blot analysis showed that the cocoa beans contained an allergenic protein of ~28 kDa. Removal of the testa strongly reduced the amount of this allergenic protein after 72 h of the pregermination treatment. Generally, the pregermination treatment increased antioxidant activity in both groups. Significant differences in the antioxidant activity were observed between groups PCB (-T) and PCB (+T). Particularly, group PCB (-T) showed high antioxidant activity at 72 h of the pregermination treatment. Thus, the combination of cocoa beans without the testa and pregermination treatment (72 h) seems to be the optimal method for production of hypoallergenic cocoa beans rich in antioxidants for patients with allergic disorders.