• Reproductive and Developmental Biology
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• v.31 no.2
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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.

• Analytical Science and Technology
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• v.18 no.6
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• pp.559-567
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• 2005

For the analytical method of PCDD/Fs in blood, which have been issued recently, SRM 1589a of NIST(National Institute of Standards and Technology) was used and a practical analytical method of PCDD/Fs in blood was presented through comparison of methods of Canada and Japan. The proposed method used alkali-digestion extraction for removal of the lipid effectively using two capillary columns. The limit of quantification of TeCDD/DF and PeCDD/DF was 1 pg/g-lipid, HxCDD/DF and HpCDD/DF was 2 pg/g-lipid, OCDD/DF was 4 pg/g-lipid. With consideration the range of detected concentration, calibration standards were presented as $CS_L$(0.1~1), $CS_1/2$(0.25~2.5), $CS_1$(0.5~5.0), $CS_2$(2~20), $CS_3$(10~100).

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.4
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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.

• Food Science of Animal Resources
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• v.37 no.1
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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.

• Journal of The Korean Society of Clinical Toxicology
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• v.1 no.1
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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.

• Food Science of Animal Resources
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• v.35 no.4
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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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• v.38 no.6
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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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• v.15 no.2
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• pp.150-155
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• 2000

Antioxidative activity of c비ture of Streptomyces sp. BH-405 was investigated. After removal of pellets of Streptomyces sp. B BH-405, antioxidative substances were is미ated and suc$\infty$sively purified from culture of Streptomyces sp. BH-405 by by thin | layer chromatography $\pi$LC) or silica gel column chromatography. The fraction 3 obtained from ethylether fractionation of the C culture appeared highest level of anti oxidative activity as determined by thiocyanate method. Band 2 obtained by further P purification of this fraction showed higher anti oxidation level than that of same concentration of dl- $\alpha$ -tocopherol, butylated h hydroxy anisole (BHA). The band 2 showed higher rate of 1, 1.diphenyl 2-picrylhydrazyl (DPPH) decolorization than dl-$\alpha$-tocopherol. In the rat liver microsomes, band 2 rapidly inhibited lipid peroxidation which was initiated enzymatically by r reduced nicotinamide adenine dinucleotide phosphate (NADPH) or non-enzymatically by Fenton’s reagent. Band 2 inhibited on | lipid peroxidation of mitochondria or the linoleic acid hydro peroxide induced peroxidation system. It is concluded that band 2 obtained by fractionation of Streptomyces sp. BH-405 cultivation contained antioxidants with the capacity to inhibit oxidative m modification.

• Journal of Food Hygiene and Safety
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• v.33 no.5
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• pp.354-360
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• 2018

This study presents a method validation for extraction and quantitative analysis of trichothecene mycotoxins in nuts based on quick, easy, cheap, effective, rugged, and safe (QuEChERS) approach for extraction and enhanced matrix removal (EMR)-lipid-disperive-SPE (d-SPE) cleanup method, with detection and quantification by high-performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) in positive- and negative-ion modes. Linearity, precision, and accuracy were validated for LC-MS/MS methods. Results obtained with LC-MS/MS were linear, with correlation coefficient ($R^2$) of 0.998. Limits of detection and quantification for mycotoxins were $0.41-3.57{\mu}g/kg$ and $1.23-10.82{\mu}g/kg$, respectively. Intra- and inter-day precisions (RSD, %) were 0.40-8.44% and 1.93-12.46%, respectively. Results indicated to be rapidly and accurately identifying trichothecene mycotoxins and may be used as a suitable safety management method for nuts and nuts related commodities.

• Journal of Plant Biotechnology
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• v.42 no.2
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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.