• Title/Summary/Keyword: Chromogenic

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Establishment of the Spectral Correction Matrix and Simultaneous Determination of Ion and Zic with 1-(6-Benzothiazolylazo)-2-Naphthol as Chromogenic Agent.

  • Geo, Hong Wen.;Li, Cheng Yu.;Ye, Qing Song.
    • Bulletin of the Korean Chemical Society
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    • v.22 no.6
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    • pp.565-569
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    • 2001
  • The spectral correction matrix has been established to study the simultaneous complextions between metals and a ligand, and it eliminated the mutual influence among the complexes. The two sensitive reations between iron(Ⅱ) and 1-(6-bromo-2-benzothiazolylazo)-2-naphthol(BBTAN) and zinc (Ⅱ) and BBTAN have been investigated and applied to the simultaneous determination of Zn and Fe at pH 8.5 in the presence of triton x-100. This method gave the simples determination of the characteristic constants of the complexes. For the analysis of natural water, foods and other samoles, the recoveries of Fe and Zn were between 92 and 109% and between 91 and 104%, and their relative standard deviations were less than 5.5 and 11%, respectively.

A Collaborative Study to Establish a Korea National Biological Standard for Antithrombin III Concentrate

  • Kang, Hye-Na;Oh, Ho-Jung;Kim, Soon-Nam;Hong, Choong-Man;Lee, Ki-Hong;Yoo, Si-Hyung;Shin, In-Soo;Choi, Seung-Eun;Lee, Seok-Ho
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.149.2-149.2
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    • 2003
  • We have carried out a collaborative study to evaluate a candidate preparation of antithrombin III concentrate whether it is suitable to serve as a Korea National Biological Standard. Three National Control Laboratories and three manufacturers participated in this study. The potency of this candidate preparation was determined by using a heparin cofactor chromogenic method described in the Minimum Requirements for Biological Products and the European Phamacopoeia. (omitted)

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Study on Overcoming Interference Factor by Automatic Synthesizer in Endotoxin Test (내독소 검사에서 자동합성장치에 따른 간섭요인 극복에 대한 연구)

  • Kim, Dong Il;Kim, Si Hwal;Chi, Yong Gi;Seok, Jae Dong
    • The Korean Journal of Nuclear Medicine Technology
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    • v.16 no.2
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    • pp.3-6
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    • 2012
  • Purpose : Samsung medical ceter shall find a cause of the interference factor and suggest a solution for it. Materials and Methods : A sample of $^{18}F$-FDG, radioactive pharmaceuticals produced by TRACERlab MX and FASTlab synthesizer. Gel-clot method uses Positive control tube and single test tube. Kinetic chromogenic method uses ENDOSAFE-PTS produced by Charles River. Results : According to Gel clot method of Endotoxin Tests at FASTlab, both turbidity and viscosity increased at 40-fold dilution and Gel clot was detected. In case of TRACERlab MX, Gel clot was detected in most of samples but intermittently not in a few of them. When using ENDOSAFE-PTS, sample CV (Coefficient of Variation) of FASTlab is 0% at all dilution rates whereas spike CV is 0% at 1-fold dilution, 0~35% at 10-fold, 3.6~12.9% at 20-fold, 5.2~7.1% at 30-fold, 1.1~17.4% at 40-fold, spike recovery; 0% at one-fold, 25 ~ 58% at 10-fold, 50 ~ 86% at 20-fold, 70~92% at 30-fold, and 75~120% at 40-fold. Sample CV of TRACERlab MX, is 0% at all dilution rates whereas spike CV is 1.4~4.8% at one-fold dilution, 0.6~19.9% at 10-fold, spike recovery; 35~72% at one-fold dilution and 77~107% at 10-fold. Conclusion : Gel clot does not seem to occur probably to H3PO4 which engages in bonding with Mg2+ion contributing gelation inside PCT. Dilution which is identical to reducing the amount of H3PO4, could remove interfering effects accordingly. Spike recovery was obtained within 70~150% - recommended values of supplier - at 40-fold dilution even in kinetic chromogenic method.

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Perspective on Rapid and Selective Method for Detecting Microbiology in Dairy Industry: A Review (낙농산업에 필요한 미생물 검사방법과 전망: 총설)

  • Chon, Jung-Whan;Kim, Hyun-Sook;Kim, Hong-Seok;Kim, Dong-Hyeon;Song, Kwang-Young;Yim, Jin-Hyuk;Choi, Dasom;Lim, Jong-Soo;Jeong, Dong-Gwan;Kim, Soo-Ki;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.2
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    • pp.119-127
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    • 2015
  • To date, detection of microbial populations in dairy products has been performed using culture media, which is a time-consuming and laborious method. The recently developed chromogenic media could be more rapid and specific than classical culture media. However, the newly developed molecular-based technology can detect microbial populations with greater rapidity and sensitivity than the classical method involving culture media and chromogenic media. This molecular-based technology could provide various options for monitoring the characterization of different states of bacteria and cells. Thus, it could help upgrade the processing system of the dairy industry so as to maintain the safety and quality of dairy foods. Among the various newly developed molecular-based technologies, flow cytometry can potentially be used for monitoring microbiological populations in the dairy industry if official international standards are available for this purpose. When omics technology would have biomarker identification, it could be regarded as the rapid and sensitive analytical methods. Methods based on PCR, which has become a basic technique in microbiological research, can be developed and validated as alternative methods for quantification of dairy microorganisms. This review discusses methods for monitoring microbiological populations in dairy foods and the limitations of these studies, as well as the need for further research on such methods in the dairy industry.

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Evaluation of Selective Media for Isolation of Foodborne Bacteria (주요 식중독균 분리용 선택배지의 타당성 연구)

  • Jo, Seo-Hee;Ha, Ji-Hyoung;Kim, Keun-Sung;Shim, Young-Hwan;Kwon, Ki-Sung;Han, Jeong-A;Hwang, In-Gyun;Ha, Sang-Do;Oh, Deog-Hwan
    • Journal of Food Hygiene and Safety
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    • v.22 no.4
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    • pp.388-394
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    • 2007
  • This study was conducted to evaluate the selective media listed in currently available Food Code in Korea. The 29 different types of media of five different types of foodborne bacteria including Salmonella spp., Bacillus cereus, Listeria monocytogenes, Vibrio parahaemolyticus, Staphylococcus aureus were tested in the broth and food. The recovery test for five different types of foodborne bacteria was performed in the artificially inoculated into chicken, rice, pork and mackerel. There was no significant differences in isolation capabilities among twenty nine different types of isolation selective media for five different types of foodborne bacteria in broth condition, while there was significantly a little differences in isolation capabilities among those on foods (P<0.05). The higher number of foodborne pathogens were isolated from conventional selective media approved in Food Code than newly developed selective media such as chromogenic media. This results suggest that there was differences of selectivities among currently available isolation selective media in many countries and further studies are needed to be approved by Korean Food and Drug Administration.

Evaluation of the Effects of Interfering Factors on the Bacterial Endotoxin Testing of Radiopharmaceuticals (방사성의약품의 박테리아 엔도톡신 시험에서 반응간섭인자들의 영향에 대한 평가)

  • Jun Young PARK
    • Korean Journal of Clinical Laboratory Science
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    • v.56 no.2
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    • pp.171-180
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    • 2024
  • The endotoxin test is based on the reaction between Limulus Amebocyte Lysate (LAL) and the lipopolysaccharides of Gram-negative bacteria. In this study, we sought to identify factors that interfere with the LAL testing of radiopharmaceuticals and evaluated acceptable ranges. A gel-clot LAL test and a chromogenic LAL test were used as endotoxin tests. We compared the performances of the Endosafe LAL and recombinant Endosafe Recombinant Cascade Reagent (rCR) cartridges for the chromogenic test. The factors that interfered with 68Ga-DOTATOC injection were pH, 4-(2-hydroxyethyl) piperazine-1-ethanesulfonic acid (HEPES) buffer, and organic solvents, especially ethanol. However, interference by these factors was overcome by diluting the 68Ga-DOTATOC injection tenfold. In addition, no interference was observed at pH values between 4 and 8, at a HEPES concentration of 2,000 ㎍/mL, or an ethanol concentration of <1%. Furthermore, results showed that interfering factors had similar effects on the performances of the Endosafe LAL and Endosafe rCR cartridges. The results of this study are expected to be useful for evaluating factors that interfere with the endotoxin testing of new radiopharmaceuticals.

Thermostable ${\beta}$-Glycosidase-CBD Fusion Protein for Biochemical Analysis of Cotton Scouring Efficiency

  • Ha, Jae-Seok;Lee, Young-Mi;Choi, Su-Lim;Song, Jae-Jun;Shin, Chul-Soo;Kim, Ju-Hea;Lee, Seung-Goo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.443-448
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    • 2008
  • Multidomain proteins for the biochemical analysis of the scouring efficiency of cotton fabrics were constructed by the fusion of a reporter moiety in the N-terminal and the cellulose binding domain (CBD) in the C-terminal. Based on the specific binding of the CBD of Cellulomonas fimi exoglucanase (Cex) to crystalline cellulose (Avicel), the reporter protein is guided to the cellulose fibers that are increasingly exposed as the scouring process proceeds. Among the tested reporter proteins, a thermostable ${\beta}$-glycosidase (BglA) from Thermus caldophilus was found to be most appropriate, showing a higher applicability and stability than GFP, DsRed2, or a tetrameric ${\beta}$-glycosidase (GUS) from Escherichia coli, which were precipitated more seriously during the expression and purification steps. When cotton fabrics with different scouring levels were treated with the BglA-CBD and incubated with X-Gal as the chromogenic substrate, an indigo color became visible within 2 h, and the color depth changed according to the conditions and extent of the scouring.

Stimulation of an Esterase Activity of Thrombin by Dequalinium and Its Relationship with Blood Coagulation

  • Paik, Seung-R.;Kim, Do-Hyung;Chang, Chung-Soon
    • BMB Reports
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    • v.29 no.3
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    • pp.225-229
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    • 1996
  • Effects on thrombin by an amphipathic cation, dequalinium, which has been recognized as an anticarcinoma agent were investigated with small chromogenic substrates such as Na-benzoyl-DL-argininep-nitroanilide (BApNA), H-D-phenylalanyl-L-pipecoyl-L-arginine-p-nitroanilide (S-2238), and Na-p-tosyl-L-arginine methyl ester (TAME). Among them, only TAME hydrolysis due to an esterase activity of the enzyme was significantly activated to 81% at 20 ${\mu}M$ dequalinium in the absence of NaCl. This stimulation became even higher in the presence of 0.2 M NaCl to 3.5-fold at 60 ${\mu}M$ dequalinium. This specific activation of thrombin was well correlated with the results of in vitro coagulation tests measuring the activated partial thromboplastin time (APTT) and the prothrombin time (PT) It is pertinent. therefore, to suggest that the esterase activity should be examined in addition to the effects on 5-2238 hydrolysis when especially any regulators not directed to an active site of thrombin need to be studied. We also expect that dequalinium could be a useful tool for studying structure-function relationship of thrombin and blood coagulation.

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Purification and Characterization of a Novel Serine Protease with Fibrinolytic Activity from Tenodera sinensis (Chinese Mantis) Egg Cases

  • Cho, So-Yean;Hahn, Bum-Soo;Kim, Yeong-Shik
    • BMB Reports
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    • v.32 no.6
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    • pp.579-584
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    • 1999
  • Mantis egg fibrolase (MEF-3) was purified from the egg cases of Tenodera sinensis using ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, DEAE Affi-Gel blue gel affinity chromatogragphy, and MONO-Q anion-exchange chromatography. This protease had a molecular weight of 35,600 Da as determined by SDS-polyacrylamide gel electrophoresis under reducing conditions and its isoelectric point was 6.0. The N-terminal amino acids sequence was Ala-Thr-Gln-Asp-Asp-Ala-Pro-Pro-Gly-Leu-Ala-Arg-Arg. This sequence was 80% homologous to the serine protease from Tritirachium album. MEF-3 readily digested the ${\alpha}$-and ${\beta}$-chains of fibrinogen and more slowly the ${\gamma}$-chains. It showed strong proteolytic and fibrinolytic activities. Phenylmethanesulfonyl fluoride and chymostatin inhibited its proteolytic activity, while EDTA, EGTA, cysteine, ${\beta}$-mercaptoethanol, elastinal, tosyl-lysine chloromethylketone, and tosyl-amido-2-phenylethyl chloromethyl ketone did not affect its proteolytic activity. Among the chromogenic protease substrates, the most sensitive one to the hydrolysis of MEF-3 was benzoyl-Phe-Val-Arg-p-nitroanilide. Based on these experimental results, we speculated that MEF-3 is a serine protease with a strong fibrin(ogen)olytic activity.

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An Anticoagulant/Fibrinolytic Protease from Lumbricus rubellus

  • Jeon, Ok-Hee;Moon, Woong-Joon;Kim, Doo-Sik
    • BMB Reports
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    • v.28 no.2
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    • pp.138-142
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    • 1995
  • An anticoagulant/fibrinolytic protease was purified to homogeneity from the earthworm Lumbricus rubellus. The protein was a single chain glycoprotein of 32 kDa that exhibited strong proteolytic activity on human thrombin and fibrin clots. Proteolytic degradation of these plasma proteins by the purified enzyme occurred at a neutral pH range. Among several human plasma proteins tested as possible substrates for the protease reaction, the 32 kDa enzyme specifically hydrolyzed both thrombin and fibrin polymers without affecting other proteins, such as serum albumin, immunoglobulin, and hemoglobin. Treatment of the purified enzyme at neutral pH with either phenylmethylsulfonylfluoride or soybean trypsin inhibitor resulted in a loss of catalytic activity. The enzyme hydrolyzed the chromogenic substrate H-D-Phe-L-Pipecolyl-L-Arg-p-nitroanilide with a $K_m$ value of 1.1 ${\mu}M$ at a neutral pH. These results suggest that the anticoagulant/fibrinolytic enzyme from Lumbricus rubellus is a member of the serine protease family having a trypsin-like active site, and one of the potential clevage sites for the enzyme is the carbonyl side of arginine residues in polypeptide chains.

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