• Title/Summary/Keyword: ethyl sulfate

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The Effect of Various Methods of Cross-linking in Type I Collagen Scaffold on Cartilage Regeneration (I형 콜라겐지지체의 다양한 가교처리 방법이 연골막성 연골재생에 미치는 영향)

  • Son, Dae Gu;Lim, Joong Jae;Sohn, Kyounghee;Yang, Eun Kyung
    • Archives of Plastic Surgery
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    • v.33 no.6
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    • pp.723-731
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    • 2006
  • Purpose: Collagen is the principal structural biomolecule in cartilage extracellular matrix, which makes it a logical target for cartilage engineering. In this study, porous type I collagen scaffolds were cross-linked using dehydrothermal(DHT) treatment and/or 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide(EDC), in the presence and absence of chondroitin-6-sulfate(CS) for cartilage regeneration. Methods: Cartilage defects were created in the proximal part of the ear of New Zealand rabbits. Four types of scaffolds(n=4) were inserted. The types included DHT cross-linked(Group 1), DHT and EDC cross- linked(Group 2), CS added DHT cross-linked(Group 3), and CS added DHT and EDC cross-linked(Group 4). Histomorphometric analysis and cartilage-specific gene expression of the reconstructed tissues were evaluated respectively 4, 8, and 12 weeks after implantation. Results: The largest quantity of regenerated cartilage was found in DHT cross-linked groups 1 and 3 in the 8th week and then decreased in the 12th week, while calcification increased. Calcification was observed from the 8th week and the area increased in the 12th week. Group 4 was treated with EDC cross-linking and CS, and the matrix did not degrade in the 12th week. Cartilage-specific type II collagen mRNA expression increased with time in all groups. Conclusion: CS did not increase chondrogenesis in all groups. EDC cross-linking may prevent chondrocyte infiltration from the perichondrium into the collagen scaffold.

Separation and Composition of Sesame Meal Protein (참깨박(粕) 단백질(蛋白質)의 분리(分離)와 조성(組成))

  • Kim, Jun-Pyong;Shim, Woo-Man;Kim, Chong-Ik
    • Applied Biological Chemistry
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    • v.23 no.1
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    • pp.14-22
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    • 1980
  • White and black sesame produced in Korea were defatted with ethyl ether or n-hexane. Defatted sesame meal was extracted with water and salt solution, and protein extraction was precipitated at various pH 1 through 12, with trichloro acetic acid (TCA), tannic acid and ammonium sulfate, respectively. Protein was purified by Sephadex A-25, G-75, G-100 and G-200, and identified its protein fraction by polyacrylamide gel electrophoresis. Amino acids composition of protein in white sesame was analyzed by automatic amino acid analyzer. Protein contents of white sesame, black sesame and sesame meal are 20.5%, 19.2%, and 44.7%, respectively. n-Hexane was the most suitable solvent for extraction of oil from sesame. Crude protein precipitation was better in higher pH. The protein extraction was more effective with the solution containing sodium chloride tinder the pH 8. Globulin in total protein was high and prolamin was less than in other cereal proteins. Glutamic acid contents of white sesame and sesame globulin were 17.1%, and 20%, respectively. Both proteins contained relatively high levels of essential amino acids. 12-13 bands were found in water soluble protein and 2 bands in salt soluble protein were detected by the disc gel electrophoresis, and were identified in both of white and black sesame. The salt soluble protein of white sesame could be purified by Sephadee G-100 and G-200.

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A Study on the Emulsion Polymerization of Methyl Methacrylate (Methyl Methacrylate의 Emulsion Polymerization에 關한 硏究)

  • Lee, Hyung-Kyoo;Min, Tae-Ik
    • Journal of the Korean Chemical Society
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    • v.12 no.1
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    • pp.4-11
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    • 1968
  • With the selected emulsifiers for the emulsion polymerization of methyl methacrylate, the HLB of the emulsifier in the reaction system has been studied on the effect of the ratio of tetra sodium-N-(1,2-dicarboxy ethyl)-N-octadecyl sulfosuccinamate(Aerosol 22) to polyethylene glycol nonyl phenyl ether (Noigen EA 160), and also sodium lauryl sulfate(Quolac EX-UB), Disodium-N-octadecyl sulfosuccinamate (Aerosol 18) and Aerosol 22 as emulsifiers having various hydrophilic groups in the molecules have been studied. Results are as follows; 1) The viscosity of the emulsions and the molecular weight of the polymers have maximum values at a constant HLB value of emulsifiers, but their stabilities show minimum point at the value with the titration with the three kinds of mono, bi, tri-valent electrolytes. These results are agreed on the theory of Greth & Wilson in which the properties of polymer emulsions depend upon the HLB system of emulsifiers. 2) The viscosity of the emulsions and the molecular weights of the produced polymers increase more in the case of blending of Aerosol 22 to Noigen EA-160 than of the separate using. 3) The coagulation effects of the divalent electrolytes($ex,\;Ca^{++},\;Zn^{++}$) are contrast to the effects of monovalent($ex,\;Na^+$) and trivalent($ex,\;Al^{+++}$) in the emulsions with Aerosol 18 or Aerosol 22 which have more than two hydrophilic groups. It seems that the stability of the O/W emulsions by electrolytes is directly related to the parameters of surface physical chemistry such as surface geometry of surface chemical constitution of polymer particles.

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Immunological Study of Induction to Salivary Glands the Cytochrome P450(IIE1) by Stress in Rat (백서 타액선의 스트레스 유도 Cytochrome P450 IIE1(CYPIIE1)에 관한 면역학적 연구)

  • Jin-Pyo Lee;Jung-Pyo Hong
    • Journal of Oral Medicine and Pain
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    • v.21 no.2
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    • pp.331-349
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    • 1996
  • Cytochrome P45O is an oxidase involved in oxidation of alcohol and is known to be an activator of carcinogen. The present study was performed to study the effect of alcohol and cold stress on the expression of Cytochrome P450 IIEl (CYPIIE1) In the liver and salivary glands in rats by an immunoblot analysis. Sixteen rats were divided into 4 groups; 1)rats belonging to group I were allowed to take 15%(v/v) ethyl alcohol as a drink ad libitum: 2)rats of group II were bathed in cold water for 30 sec twice a day (during the one-week experiment); 3)rats comprising group III were received alcohol and cold stress as described above; 4)rats of group IV were selected as a control. The rat were sacrificed at the end of the one-week experiment. The livers and parotid and submandibular salivary glands were removed and stored at -2$0^{\circ}C$ until use. The stored organs were homogenized for 10 sec and the supernatants were obtained by centrifugation. The proteins of the supernatants were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and subjected to Western blotting. The blotted membranes were incubated with polyclonal antibodies to CYPIIEI . The obtained results were as follows : 1. The expression of CYPIIEl was apparently negative in the liver and salivary glands of group IV, wheras its expression was marked in the experiment groups I, II. and III. 2. No difference in the expression of CYPIIEl in the liver and salivary glands was observed between the experiment groups I, II, and III. 3. Among the experiment groups, the expression of CYPIIE1 in the liver was much greater than in the salivary glands. The expression of CYPIIE1 in the submandibular gland was weakly positive but was greater than in the carotid gland.

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Comparative Biochemical Properties of Proteinases from the Hepatopancreas of Shrimp. -I. Purification of Protease from the Hepatopancreas of Penaeus japonicus-

  • Choi Sung-Mi;Oh Eun-Sil;Kim Doo-Sang;Pyeun Jae-Hyeung;Cho Deuk-Moon;Ahn Chang-Bum;Kim Hyeung-Rak
    • Fisheries and Aquatic Sciences
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    • v.1 no.2
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    • pp.201-208
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    • 1998
  • A protease, which had no tryptic and chymotryptic activity, was purified from the hepatopancreas of shrimp, P. japonicus, through ammonium sulfate fractionation, Q­Sepharose ionic exchange, benzamidine Sepharose 6B affinity, and Sephacryl S-100 gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 24 kDa by gel filtration and showed a single peptide band by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The protease had a low ratio of acidic to basic amino acids, which is different with pro teases from marine animals. The enzyme was partially inhibited by benzamidine, tosyl-L-lysine chioromethyl ketone (TLCK), phenylmethylsulfonyl fluoride (PMSF), soybean trypsin inhibitor (SBTI), and pepstatin. The enzyme did not have any activity against benzoyl-D,L-arginine p-nitroanilide (BAPNA) or benzoyl-L-tyrosine ethyl ester (BTEE) which is a specific substrate of trypsin and chymotrypsin, respectively. However, the enzyme showed activity forward N-CBZ-L-tyrosine p-nitrophenyl ester (CBZ-Tyr-pNE), N­CBZ-L-tryptophan p-nitrophenyl ester (CBZ-Trp-pNE), and N-CBZ-L-proline p-nitrophenyl ester (CBZ-Pro-pNE). The protease did not showed tryptic and chymotryptic activity, which was not reported in shrimp hepatopancreas.

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Study on Effective Preservation of Bovine Pericardium Using Decellulariation and ${\alpha}$-galactosidase for Eliminating Xenoreactive Antigen (이종 항원 제거를 위한 무세포화와 알파-갈락토시다아제를 이용한 효과적인 우심낭 보존 방법에 관한 연구)

  • Kim, Min-Seok;Park, Cham-Jin;Kim, Soo-Hwan;Lim, Hong-Gook;Kim, Yong-Jin
    • Journal of Chest Surgery
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    • v.43 no.6
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    • pp.576-587
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    • 2010
  • Background: Effective decellularization and fixation process is critical, in order to use xenogenic valves clinically. In the present study, we decellularized bovine pericardium using sodium dodecyl sulfate (SDS) and N-lauroyl sarcosinate, treated with $\alpha$-galactosidase, and then fixed in various manners, to find out the most effective tissue preservation & fixation procedure. Material and Method: Bovine pericardium was decellularized with SDS and N-lauroyl sarcosinate, and treated with $\alpha$-galactosidase. Both groups were fixed differently, by varying glutaraldehyde (GA) or EDC (1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide)/N-hydroxysuccinamide (NHS) treatment conditions. Thereafter, physical examination, tensile strength test, thermal stability test, cytotoxicity test, pronase test, pronase-ninhydrin test, purpald test, permeability test, compliance test, H&E staining, DNA quantification, and $\alpha$-galactose staining were carried out to each groups. Result: GA fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, EDC/NHS-GA dual fixed groups showed better physical properties and thermal stability than EDC/NHS fixed groups, and showed better thermal stability than GA fixed groups. In pronase test and pronase-ninhydrin test, GA fixed groups and EDC/NHS-GA dual fixed groups showed stronger crosslinks than EDC/NHS groups. Permeability and compliance tended to increase in EDC/NHS-GA dual fixed groups, compared to GA fixed groups. But, EDC/NHS-GA dual fixed groups had stronger tensile strength and lower cytotoxicity than GA fixed groups. Conclusion: We have verified that EDC/NHS-GA dual fixation can make effective crosslinks and lower the toxicity of GA fixation. Henceforth, we will verify if EDC/NHS-GA dual fixation can lower calcifications & tissue failure in vivo experiment.

Development of Purification Process of Recombinant Human Vascular Endotherial Growth Factor (VEGF) using Fusion Protein (융합 단백질을 이용한 재조합 인간 혈관내피세포 성장인자의 정제공정 개발)

  • Sung, Keehyun;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.55 no.3
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    • pp.369-378
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    • 2017
  • Vascular endotherial growth factor (VEGF) is a potent mitogen that stimulates vascular permeability and angiogenesis and has a potential in therapeutic applications. An industrial production method that provides high yield as well as purity is needed. Researches for various factors of mild solubilization with combination of ubiquitin fusion protein to increase solubility were carried out as well as by changing pH and denaturant concentration. Usage of pET28-a bacteral expression vector in BL21 (DE3) host cell was capable of producing approximately 14 g/L VEGF fusion protein in 20L fermentor. A purification process consisting of four chromatography steps including refolding and digestion with UBP1 resulted in mild solublization under the conditions of 2M urea and pH 10.0 due to ubiquitin fusion tag protein that increases in solubility of target protein VEGF. High yield of refolding and dimerization could be obtained between two step Ni-affinity chromatography. Multimeric and misfolded proteins and endotoxin were removed by DEAE anion exchange chromatography. Final monomers were removed from dimers by gel filtration chromatography. Characterization analysis of purified dimeric VEGF was performed using SDS-PAGE and RP-HPLC with a purity of 97%.

Column Comparison for the Separation of Ferimzone Z and E Stereoisomers and Development of Trace Residue Analysis Method in Brown Rice Using HPLC-MS/MS (컬럼 비교를 통한 Ferimzone Z 및 E 입체 이성질체의 물질 분리 및 HPLC-MS/MS를 활용한 현미 중 미량잔류분석법 개발)

  • Mun-Ju Jeong;So-Hee Kim;Hye-Ran Eun;Ye-Jin Lee;Su-Min Kim;Jae-Woon Baek;Yoon-Hee Lee;Yongho Shin
    • Korean Journal of Environmental Agriculture
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    • v.42 no.3
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    • pp.203-210
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    • 2023
  • Ferimzone Z is a fungicide for effectively controlling rice blast. Under light irradiation conditions, it undergoes a rapid conversion to its E-stereoisomer. Given the importance of isomers in risk assessments of residues in crops, an analytical method was developed for individual isomer quantification. A comparative analysis performed using two columns in HPLC-MS/MS demonstrated that the isomers were successfully separated using the Cadenza column. For the brown rice sample preparation, 5 g of the homogenized sample was saturated with 7 mL of water. The sample was then extracted with a 10 mL mixed solvent of acetonitrile and ethyl acetate (1:1, v/v) that contained 0.1% formic acid, and it was subsequently partitioned with magnesium sulfate and sodium chloride. The upper layer was purified using dSPE containing C18 and PSA sorbents. The established method was subjected to method validation, and it showed recovery rates of 90.6-98.8% (RSD ≤ 3.9%) at concentrations of 0.01, 0.1, 2 mg/kg, with a soft matrix effect (%ME) ranging from -3.1% to +6.5%. This method can be employed in monitoring studies of brown rice to determine the conversion ratio from the Z isomers to the E isomers.

Development of a Simultaneous Analytical Method for Azocyclotin, Cyhexatin, and Fenbutatin Oxide Detection in Livestock Products using the LC-MS/MS (LC-MS/MS를 이용한 축산물 중 유기주석계 농약 Azocyclotin, Cyhexatin 및 Fenbutatin oxide의 동시시험법 개발)

  • Nam Young Kim;Eun-Ji Park;So-Ra Park;Jung Mi Lee;Yong Hyun Jung;Hae Jung Yoon
    • Journal of Food Hygiene and Safety
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    • v.38 no.5
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    • pp.361-372
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    • 2023
  • Organotin pesticide is used as an acaricide in agriculture and may contaminate livestock products. This study aims to develop a rapid and straightforward analytical method for detecting organotin pesticides, specifically azocyclotin, cyhexatin, and fenbutatin oxide, in various livestock products, including beef, pork, chicken, egg, and milk, using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The extraction process involved the use of 1% acetic acid in a mixture of acetonitrile and ethyl acetate (1:1). This was followed by the addition of anhydrous magnesium sulfate (MgSO4) and anhydrous sodium chloride. The extracts were subsequently purified using octadecyl (C18) and primary secondary amine (PSA), after which the supernatant was evaporated. Organotin pesticide recovery ranged from 75.7 to 115.3%, with a coefficient of variation (CV) below 25.3%. The results meet the criteria range of the Codex guidelines (CODEX CAC/GL 40). The analytical method in this study will be invaluable for the analysis of organotin pesticides in livestock products.

Studies on the physio-chemical properties and the cultivation of oyster mushroom(Pleurotus ostreatus) (느타리버섯의 생리화학적성질(生理化學的性質) 및 재배(栽培)에 관(關)한 연구(硏究))

  • Hong, Jai-Sik
    • Applied Biological Chemistry
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    • v.21 no.3
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    • pp.150-184
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    • 1978
  • Nutritional characteristics and physio-chemical properties of mycelial growth and fruitbody formation of oyster mushroom(Pleurotus ostreatus)in synthetic media, the curtural condition for the commerical production in the rice straw and poplar sawdust media, and the changes of the chemical components of the media and mushroom during the cultivation were investigated. The results can be summarized as follows: 1. Among the carbon sources mannitol and sucrose gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while lactose and rhamnose gave no mycelial growth. Also, citric acid, succinic acid, ethyl alcohol and glycerol gave poor fruit-body formation, and acetic acid, formic acid, fumaric acid, n-butyl alcohol, n-propyl alcohol and iso-butyl alcohol inhibited mycelial growth. 2. Among the nitrogen sources peptone gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while D,L-alanine, asparatic acid, glycine and serine gave very poor fruit-body formation, and nitrite nitrogens, L-tryptophan and L-tyrosine inhibited mycelial growth. Inorganic nitrogens and amino acids added to peptone were effective for fruit-body growth, and thus addition of ammonium sulfate, ammonium tartarate, D,L-alanine and L-leucine resulted in about 10% increase fruit-body yield. L-asparic acid about 15%, L-arginine about 20%, L-glutamic acid, and L-lysine about 25%. 3. At C/N ratio of 15.23 fruit-body formation was fast, but the yield decreased, and at C/N ratio of 11.42 fruit-body formation was slow, but the yield increased. Also, at the same C/N ratio the higher the concentration of mannitol and petone, the higher yield was produced. Thus, from the view point of both yield of fruit-body and time required for fruiting the optimum C/N ratio would be 30. 46. 4. Thiamine, potassium dihydrogen phosphate and magnecium sulfate at the concentration of $50{\mu}g%$. 0.2% and 0.02-0.03%, respectively, gave excellent mycelial and fruit-body growth. Among the micronutrients ferrous sulfate, zinc sulfate and manganese sulfate showed synergetic growth promoting effect but lack of manganese resulted in a little reduction in mycelial and fruit-body growth. The optimum concentrati on of each these nutrients was 0.02mg%. 5. Cytosine and indole acetic acid at 0.2-1mg% and 0.01mg%, respectively, increased amount of mycelia, but had no effect on yield of fruit-body. The other purine and pyrimidine bases and plant hormones also had no effect on mycelial and fruit-belly yield. 6. Illumination inhibited mycelial growth, but illumination during the latter part of vegetative growth induced primordia formation. The optimum light intensity and exposure time was 100 to 500 lux and 6-12 hours per day, respectively. Higher intensity of light was injurous, and in darkness only vegetative growth without primordia formation was continued. 7. The optimum temperature for mycelial growth was $25^{\circ}C$ and for fruit-body formation 10 to $15^{\circi}C$. The optimum pH range was from 5.0 to 6.5. The most excellent fry it-body formation were produced from the mycelium grown for 7 to 10 days. The lesser the volume of media, the more rapid the formation of fruit-body; and the lower the yield of fruit-body; and the more the volume of media, the slower the formation of fruit-body, and the higher the yield of fruit-body. The primordia formation was inhibited by $CO_2$. 8. The optimum moisture content for mycelial growth was over 70% in the bottle media of rice straw and poplar sawdust. 10% addition of rice bran to the media exhibited excellent mycelial growth and fruit-body formation, and the addition of calciumcarbonate alone was effective, but the addition of calcium carbonate was ineffective in the presence of rice bran. 9. In the cultivation experiments the total yield of mushroom from the rice straw media was $14.99kg/m^2$, and from the sawdust media $6.52kg/m^2$, 90% of which was produced from the first and second cropping period. The total yield from the rice straw media was about 2.3 times as high as that from the sawdust media. 10. Among the chemical components of the media little change was observed in the content of ash on the dry weight basis, and organic matter content decreased as the cultivation progressed. Moisture content, which was about 79% at the time of spawning, decreased a little during the period of mycelial propagation, after which no change was observed. 11. During the period from spawning to the fourth cropping about 16.7% of the dry matter, about 19.3% of organic matter, and about 40% of nitrogen were lost from the rice straw media; about 7.5% of dry mallet, about 7.6% of organic matter, and about 20% of nitrogen were lost from the sawdust media. For the production of 1kg of mushroom about 232g of organic matter and about 7.0g of nitrogen were consumed from the rice straw media; about 235g of organic matter and about 6.8g of nitrogen were consumed from the sawdust media, 1㎏ of mushroom from either of media contains 82.4 and 82.3g of organic matter and 5.6 and 5.4g of nitrogen, respectively. 12. Total nitrogen content of the two media decreased gradually as the cultivation progressed, and total loss of insoluble nitrogen was greater than that of soluble nitrogen. Content of amino nitrogen continued to increase up to the third cropping time, after which it decreased. 13. In the rice straw media 28.0 and 13.8% of the total pentosan and ${\alpha}$-cellulose, respectively, lost during the whole cultivation period was lost during the period of mycelial growth; in the sawdust media 24.1 and 11.9% of the total pentosan and ${\alpha}$-cellulose, respectively, was lost during the period of mycelial growth. Lignin content in the media began to decrease slightly from the second cropping time, while the content of reduced sugar, trehalose and mannitol continued to increase. C/N ratio of the rice straw media decreased from 33.2 at spawining to 30.0 at ending; that of the sawdust media decreased from 61.3 to 60.0. 14. In both media phosphorus, potassium, manganese and zinc decreased, at magnesium, calcium and copper showed irregular changes, and iron had a tendency to be increased. 15. Enzyme activities are much higher in the rice straw media than in the sawdust media. CMC saccharifying and liquefying activity gradually increased from after mycelial propagation to the second cropping, after which it decreased in both media. Xylanase activity rapidly and greatly increased during the second cropping period rather than the first period. At the start of the third cropping period the activity decreased rapidly in the rice straw media, which was not observed in the sawdust media. Protease activity was highest after mycelial propagation, after which it gradually decreased. The pH of the rice straw media decreased from 6.3 at spawning to 5.0 after fourth cropping; that of the sawdust media decreased from 5.7 to 4.9. 16. The contents of all the components except crude fibre of the mushroom from the rice straw media were higher than those from the sawdust media. Little change was observed in the content of the components of mushroom cropped from the first to the third period, but slight decrease was noticed at the fourth cropping.

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