• Title/Summary/Keyword: enzyme solutions

Search Result 103, Processing Time 0.022 seconds

Differential Expression of EGFR Protein by Immunohistochemical Staining Methods and the Relationship Between the Degree of EGFR Protein Expression and EGFR Gene Mutation (면역조직화학적 염색 방법에 따른 상피세포 성장 수용체 단백(EGFR)의 발현정도의 차이 및 EGFR의 발현정도와 EGFR 유전자의 돌연변이와의 상관관계에 대한 고찰)

  • Yoon, In-Sook;Kim, Keuk-Jun;Lee, Eun-Hwa;Seok, Sang-Hee;Kim, Sang-Hee;Kim, Hyun-Yong;Song, Ho-Jung;Lee, Tae-Jong
    • Korean Journal of Clinical Laboratory Science
    • /
    • v.39 no.3
    • /
    • pp.217-222
    • /
    • 2007
  • In the last 5 years the Epidermal Growth Factor Receptor (EGFR) has emerged as one of the most important targets for drug development in oncology. Monoclonal antibodies targeting the external domain of EGFR have been shown to have clinical benefits in colorectal and head and neck cancer when combined with chemotherapy and/or radiation. Also the targeting of the epithelial growth factor receptor (EGFR) kinase domain using the closely related inhibitors gefitinib and erlotinib has generally been ineffective against solid tumors, many of which over express the receptor. We found that there were some differential expressions according to primary antibodies of the EGFR protein which being used as one of the histological tumor markers for non-small cell lung cancer (NSCLC). We also found that there are some differential expressions according to antibodies, the pH of the antigen retrieval (AR) buffer solutions and kinds of enzymes. There were some differential expressions according to the secondary antibodies and the detection systems. We analyzed the correlations between the immunohistochemical expressions of the EGFR protein and the gene mutations of the EGFR. The differences between automatic stainers and manual staining methods were also evaluated.

  • PDF

Photolithographic Fabrication of Poly(Ethylene Glycol) Microstructures for Hydrogel-based Microreactors and Spatially Addressed Microarrays

  • Baek, Taek-Jin;Kim, Nam-Hyun;Choo, Jae-Bum;Lee, Eun-Kyu;Seong, Gi-Hun
    • Journal of Microbiology and Biotechnology
    • /
    • v.17 no.11
    • /
    • pp.1826-1832
    • /
    • 2007
  • We describe the fabrication of poly(ethylene glycol) diacrylate (PEG-DA) hydrogel microstructures with a high aspect ratio and the use of hydrogel microstructures containing the enzyme ${\beta}$-galactosidase (${\beta}$-Gal) or glucose oxidase (GOx)/horseradish peroxidase (HRP) as biosensing components for the simultaneous detection of multiple analytes. The diameters of the hydrogel microstructures were almost the same at the top and at the bottom, indicating that no differential curing occurred through the thickness of the hydrogel microstructure. Using the hydrogel microstructures as microreactors, ${\beta}$-Gal or GOx/HRP was trapped in the hydrogel array, and the time-dependent fluorescence intensities of the hydrogel array were investigated to determine the dynamic uptake of substrates into the PEG-DA hydrogel. The time required to reach steady-state fluorescence by glucose diffusing into the hydrogel and its enzymatic reactions with GOx and HRP was half the time required for resorufin ${\beta}$-D-galactopyranoside (RGB) when used as the substrate for ${\beta}$-Gal. Spatially addressed hydrogel microarrays containing different enzymes were micropatterned for the simultaneous detection of multiple analytes, and glucose and RGB solutions were incubated as substrates. These results indicate that there was no cross-talk between the ${\beta}$-Gal-immobilizing hydrogel micropatches and the GOx/HRP-immobilizing micropatches.

Determination of Ovalbumin in Processed Foods by Immunological Methods

  • Seo, Ji-Hyun;Lee, Ju-Woon;Kang, Sin-Bok;Lee, Ha-Baik;Yook, Hong-Sun;Kim, Mee-Ree;Kim, Kee-Hyuk;Byun, Myung-Woo
    • Preventive Nutrition and Food Science
    • /
    • v.7 no.4
    • /
    • pp.373-377
    • /
    • 2002
  • Allergens in processed foods may place persons with food allergies at significant risk when the labels do not Provide sufficient warnings or identification of high-risk ingredients. Because egg proteins are common food allergens, this study was carried out to identify hen's egg albumin (ovalbumin, OVA) in five commercially processed foods containing egg (custayd, cookie and pasta), and chicken meat (sausage and meatball) by immunological methods using commercially produced murine monoclonal immunoglobulin G (M-IgG), immunoblotting and enzyme linked immunosorbent assay (ELISA). Sample buffer with chelating and reducing agents was prepared and used for the preparation of the protein fractions from the foods. Most bands in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) profile (5~15% gradient gel) presented at 75 kDa below. OVA (43 kDa) in the sample lanes could not be visually observed on the gel. However, OVA in solutions prepared from custard and cookie could be detected by M-IgG, but were not detected in sausage and pasta. OVA in all samples could be quantitatively determined by the equation obtained from the standard curve by ELISA. Cookie and custard containing egg white and egg, respectively, contained very high concentrations of OVA. OVA in the other products were present in relatively low concentrations, but sufficiently high to pose possible risk of allergy, ELISA is a very sensitive and precise method for the identification and quantification of allergens in food products including allergy-inducible materials.

Algorithm for Integral Method for Photocurrent Measurement of pH Variations Using Multichannel LAPS (다채널 LAPS용 pH 변화량 검출을 위한 적분 방식의 알고리듬 제안)

  • Bae, Sang-Kon;Lee, Sung-Ha;Kang, Shin-Won;Cho, Jin-Ho
    • Journal of the Institute of Electronics Engineers of Korea SC
    • /
    • v.38 no.6
    • /
    • pp.65-75
    • /
    • 2001
  • We proposed the detection method of pH variations by integrating a photocurrent characteristic curve, instead of finding an inflection point by differentiating it in LAPS system. By a simulation of the performance of the proposed method, we verified that it had 80 and 1000 times higher sensitivity and resolution than a conventional method. Then, with the implemented system based on the simulation results, we measured a pH variation which was given rise to a potential change on the LAPS surface exposed to 2-0.03125[mg/ml] enzyme solutions. As results, we observed that the proposed method has a higher sensitivity and resolution of 3.76-0.08[pH/min] pH variations than 3.79-0.27[pH/min] for conventional method with same samples.

  • PDF

Changes of Myocardial Ultrastructure During Open Heart Surgery (개심수술중 심근 미세구조의 변화)

  • Hur, Jin;Kong, Joon-Hyuk;Lee, Jong-Tae
    • Journal of Chest Surgery
    • /
    • v.33 no.9
    • /
    • pp.707-715
    • /
    • 2000
  • Background: Cold blood cardioplegic solution has been used to protect myocardium during open heart surgery with the hypothesis stating that it provides more oxygen supply to myocardium compared to crystalloid caridoplegic solution. We repeatedly infused cold blood cardioplegic solution to achieve myocardial protection. We biopsied a small portion of papillary muscle of patients with mitral valve replacement or double valve replacement during aortic cross-clamp time and evaluated the method of myocardial protection through the observation of changes in ultrastructure. We then analysed the relationship between changes in ultrasructure and peak postoperative CK-MB value and SGOT value. Material and method: We report observation on changes of myocardial ultrastructure, postoperative CK-MB and SGOT, and electrocardiogram in 31 patients who underwent cardiac operation. There were 11 males and 20 females, and they ranging in age from 28 to 69 years(mean score was 2.08$\pm$0.560, it was 2.37$\pm$0.558 at 40 minutes, and it was 2.36$\pm$0.523 at 70minutes. Mitochondrial score increased significant at 40 minutes. Mean value of postoperative peak CK-MB and SGOT were 37.3$\pm$17.061IU, 144.5$\pm$125.5IU respectively. We were not able to find any new Q were in EKG after the operation. There was no significant relationship between myocardium mitochondrial score and mean value of postoperative peak CK-MB and SGOT. Conclusion: In conclusion, with this study the cold blood cardioplegic solution was incomplete in preserving ultrastructure of myocardium even with satisfactory results in serum enzyme and EKG evaluation.

  • PDF

Protective Effect of Selected Amino Acids and Food Extracts on Ethanol Toxicity Decrement in Rat Liver (일부 아미노산과 식품 추출물의 에탄올 간독성에 대한 보호효과)

  • Lee, Ja-Hyun;Kim, N.K.;Lee, Do-Youn;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
    • /
    • v.31 no.3
    • /
    • pp.802-808
    • /
    • 1999
  • An rat liver enzyme test was carried out in order to investigate preventing effect of selested amino acids and some food extracts on ethanol induced liver toxicity in vitro. Solutions of aspartic acid, arginine, glutamic acid were prepared and treated on ethanol treated rat liver preparation. Protective effect of amino acids on lipid peroxidation was determined. Same experiments were conducted using aqueous extracts of Dried soybean sprout, Dried Alaskan pollack and Ganoderma lucidum. The TBA value indicating the lipid peroxidation decreased significantly (p<0.05) by addition of aspartate, glutamate and arginine, repectively at concentrations of $6.25{\sim}50\;{\mu}g/mL$. Similar results were observed by adding the aqueous extracts of Soybean sprout, dried Alaskan pollack and Ganoderma lucidum. The aqueous extracts added after ethanol treatment presemted more effect than added before the treatment.

  • PDF

Evaluation of Nattokinase for Antithrombotic Effect and Pharmacological Efficacy by a Biological Test and Clinical Trial (동물 및 인체시험을 통한 Nattokinase의 항응고 작용 및 섬유소 용해능 평가)

  • Kim, Jae-Bum;Yoo, Chul-Bae;Shin, Hyun-Man;Jung, Joon-Ki;Jang, Hyung-Wook
    • KSBB Journal
    • /
    • v.26 no.5
    • /
    • pp.393-399
    • /
    • 2011
  • Bacillus subtilis natto producing high level of a fibrinolytic enzyme was selected and Ultra Nattokinase$^{(R)}$ was manufactured by fermentation and purification. It was performed the evaluation of the antithrombotic effect of Ultra Nattokinase$^{(R)}$ (20,000 FU/g) with rat blood plasma. The maximum aggregation (inhibition ratio) was 71% (0%), 69% (2.8%), 62% (12.7%), 16% (77.5%) and 9% (87.3%), respectively, in the order of 0, 5, 10, 50 and 100 mg/mL of Ultra Nattokinase$^{(R)}$ solutions. Ultra Nattokinase$^{(R)}$ had antithrombotic effect, which was associated with the suppression of collagen-induced platelet aggregation. Ultra Nattokinase$^{(R)}$ in the topic of the FDP (fibrinogen degradation products) in blood coagulation tests showed a significant increasing trend. And based on the daily record of meal 39 people of ITT (what ?) group consisted with 19 people of NP (what ?) group and 20 people of PN (what ?) group except four people, two people who took vitamin K affecting the experiment and two people who took alcohol, finding to be taken Ultra Nattokinase$^{(R)}$ showed an increase in the FDP value after four weeks. In addition, FDP value of 41 people of ITT group except two people having metabolic syndrome was increased by Ultra Nattokinase$^{(R)}$.

Clinical Observations on Myocardial Protection with Cold Blood Potassium Cardioplegia According to the Duration of Ischemic Arrest (냉혈K+ 심정지약을 사용한 개심술 예에서 심정지시간에 따른 심근보호효과에 관하여)

  • Jeong, J.G.;Lee, D.H.
    • Journal of Chest Surgery
    • /
    • v.24 no.7
    • /
    • pp.647-655
    • /
    • 1991
  • The efficacy of cold blood potassium cardioplegia during periods of ischemic arrest was assessed in 88 patients undergoing open heart surgical procedures at Chonnam National University Medical School from December, 1987 to January, 1989. The purpose of this study was to determine if the aortic cross clamping time[ACCT] over 120 minutes correlated with operative mortality, incidence of postoperative ventricular tachyarrhythmias, needs of postoperative inotropic support and serum enzyme levels. The patients were divided according to aortic cross clamping time[less than 120 minutes and 120 minutes or greater]. The results were as follows: 1. The operative mortality was 3.2% in ACCT<120min group and 7.7% in ACCT>120 min group. 2. The incidence of postoperative ventricular tachyarrhythmia was 1.6% in ACCT <120min group and 11.5% in ACCT>120min group[p<0.05]. 3. The incidence of postoperative inotropic support in congenital heart disease was 13.0Fo in ACCT<120min group and 45.0%o in ACCT>120min group[p<0.05]. The incidence in acquired heart disease was 26.0% in ACCT<120min group and 40.0% in ACCT> 120min group. 4. After cardiopulmonary bypass, serum GOT, LDH, CPK and CPK - MB were elevated prominently. Children showed higher value of the enzymes examined than adults did before and after cardiopulmonary bypass. In congenital heart diseases, postoperative serum GOT, LDH, CPK and CPK - MB levels of ACCT>120min group were significantly higher than those of ACCT<120min group. Postoperative serum GOT, LDH and CPK - MB levels of ACCT>=120min group were significantly higher than those of ACCT<120min group also in acquired heart diseases. The results suggest that the myocardial protective effect with cold blood potassium cardioplegic solutions was not sufficient when the aortic cross clamping time was over 120 minutes.

  • PDF

Isolation and Identification of Korean type Streptococcus mutans (한국형 Streptococcus mutans의 분리 및 동정)

  • 현성희;장성렬;최영길
    • Korean Journal of Microbiology
    • /
    • v.27 no.3
    • /
    • pp.250-258
    • /
    • 1989
  • S. mutans known as a causative causative agent of dental caries was isolated from a carious lesion of Korean in the present study. The physiological, biochemical characteristics and polysaccharide pattern of these isolates were compated with those of four laboratory strains ; AHT(a), FA-1F(b), LM7(e), and OMZ65(g). One hundred strains of oral streptococci were isolated from dental caries sites of Korean (one male and one female). Among these, 3 strains were identified as S. mutans. These strains were able to grow on selective media MS, MST, MSP, MSP1, MSB, MSBT and were stained dark pink when sprayed with solutions of mannitol and TTC. So, these strains were called strain 108, 110, and 120, respectively. Strain 108, 110, and 120 were bacitracin resistnt. As these strains contained particularly hippurate hydrolysis enzyme, they were distinguished from laboratory strains. Apart from laboratory strains, the strain 108 was not capable of fermenting lactose, the strain 110 was not able to ferment sorbitol, inulin, melibiose, raffinose and the strain120 was incapable of fermenting inulin, raffinose. All fractions of extracellular and ecll bound polysaccharide of the strain108, 110, and 120 were consisted of more glucan than fructan. Aside from laboratory strains, the isolated strains were composed of more water-insoluble glucans related adherence on solid surface than water-soluble. According to these results, the strain108, 110, and 120 had native characteristecs of S. mutans, but they were different from laboratory strains in some characteristics. Therefore, each of them was given a name to S. mutans KHC108, KHC110, and KHC120, respcetively.

  • PDF

Trypsins from the Dark Fleshed Fish(Anchovy, Mackerel, Yellowfin Tuna and Albacore) 1. Purification and Optimal Reaction Conditions (혈합육어(멸치, 고등어, 황다랭이 및 날개다랭이)의 Trypsin 1. 정제와 반응조건)

  • 변재형;조득문;허민수
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.22 no.4
    • /
    • pp.448-457
    • /
    • 1993
  • Deterioration of fish muscle is known to occur more quickly in the dark fleshed fish than in the white fleshed fish, causing by their high intestinal proteolytic activity. Muscle degradation which suffer post-mortem autoproteolysis is affected by trypsin with its unique activation function towards other enzymes. To compare physicochemical and enzymatic properties for the trypsins of the dark fleshed fish, trypsins from the viscera of anchovy (Engraulis japonica), and the pyloric caeca of mackerel (Scomber japonicus), yellowfin tuna (Thunnus albacores) and albacore (Thunnus alalunga) were purified through ammonium sulfate fractionation, benzamidine-Sepharose 6B, DEAE-Sephadex A-50, and Sephadex G-75 chromatography Two trypsins from mackerel (designated mackerel trypsin A and mackerel trypsin B), and one each from anchovy, yellowfin tuna and albacore were isolated as electrophoretical homogeneity, The purities of anchovy trypsin, mackerel trypsin A and B, yellowfin tuna trypsin, and albacore trypsin increased to 78.1, 4.8, 9.3, 120, and 160-fold, respectively, compared to crude enzyme solutions. Molecular weights of the trypsins from the dark fleshed fish estimated by SDS-polyacrylamide electrophoresis were ranged from 22kDa to 26kDa. The trypsins contained higher amount of glycine, serine and aspartic acid, and less amount of tryptophan, methionine, lysine and tyrosine. Optimal conditions for amidotici reactions of the enzymes were pH 8.0 and 45$^{\circ}C$ for anchovy trypsin, pH 8.0 and 5$0^{\circ}C$ for mackerel trypsin A and B, pH 9.0 and 55$^{\circ}C$ for yellowfin tuna trypsin, and pH 9.0 and 5$0^{\circ}C$ for albacore trypsin. It was supposed that the habitat temperature of the dark fleshed fish is slightly connected with the optimal reaction temperature of the trypsins of the fish.

  • PDF