Science.gov

Sample records for a-421-811 purified carboxymethylcellulose

  1. 76 FR 29194 - Purified Carboxymethylcellulose From Mexico and Sweden: Revocation of Antidumping Duty Orders

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-20

    ... International Trade Administration Purified Carboxymethylcellulose From Mexico and Sweden: Revocation of... duty orders on purified carboxymethylcellulose from Mexico and Sweden. Pursuant to section 751(c) of... of the existing antidumping duty orders on purified carboxymethylcellulose from Mexico and...

  2. 75 FR 57815 - Purified Carboxymethylcellulose From Finland, Mexico, Netherlands, and Sweden

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-22

    ... COMMISSION Purified Carboxymethylcellulose From Finland, Mexico, Netherlands, and Sweden AGENCY: United... antidumping duty orders on purified carboxymethylcellulose from Finland, Mexico, Netherlands, and Sweden... antidumping duty orders on purified carboxymethylcellulose from Finland, Mexico, Netherlands, and Sweden...

  3. 77 FR 14733 - Purified Carboxymethylcellulose From Finland and the Netherlands: Extension of Time Limit for...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-13

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF COMMERCE International Trade Administration Purified Carboxymethylcellulose From Finland and the Netherlands: Extension..., inter alia, purified carboxymethylcellulose from Finland and the Netherlands covering the period July...

  4. 75 FR 39207 - Purified Carboxymethylcellulose From Finland: Extension of Time Limit for Preliminary Results of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-08

    ... International Trade Administration Purified Carboxymethylcellulose From Finland: Extension of Time Limit for... 30, 2010. See Purified Carboxymethylcellulose From Finland: Extension of Time Limit for Preliminary... Time Limits for Preliminary Results Section 751(a)(3)(A) of the Tariff Act of 1930, as amended (the...

  5. 76 FR 3159 - Purified Carboxymethylcellulose From Finland, Mexico, Netherlands, and Sweden

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-19

    ... From the Federal Register Online via the Government Publishing Office INTERNATIONAL TRADE COMMISSION Purified Carboxymethylcellulose From Finland, Mexico, Netherlands, and Sweden AGENCY: United States International Trade Commission. ACTION: Revised schedule for the subject reviews. DATES:...

  6. 75 FR 3444 - Purified Carboxymethylcellulose From Finland: Extension of Time Limit for Preliminary Results of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-01-21

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF COMMERCE International Trade Administration Purified Carboxymethylcellulose From Finland: Extension of Time Limit for Preliminary Results of Antidumping Duty Administrative Review AGENCY: Import Administration, International Trade Administration, Department...

  7. 76 FR 29191 - Purified Carboxymethylcellulose From Finland and the Netherlands: Continuation of Antidumping...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-20

    ... Antidumping Duty Orders: Purified Carboxymethylcellulose from Finland, Mexico, the Netherlands and Sweden, 70... Netherlands, and Sweden: Final Results of the Expedited First Sunset Reviews of the Antidumping Duty Orders... Finland, Mexico, Netherlands and Sweden, 76 FR 27663 (May 12, 2011), and USITC Publication 4225 (May...

  8. 75 FR 14422 - Purified Carboxymethylcellulose from Mexico: Extension of Time Limits for Preliminary Results of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-25

    ... International Trade Administration Purified Carboxymethylcellulose from Mexico: Extension of Time Limits for... Time Limits for Preliminary Results Section 751(a)(3)(A) of the Tariff Act of 1930, as amended (the.... However, if it is not practicable to complete the review within this time period, section 751(a)(3)(A)...

  9. 76 FR 27663 - Purified Carboxymethylcellulose From Finland, Mexico, Netherlands and Sweden

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-12

    ... From the Federal Register Online via the Government Publishing Office INTERNATIONAL TRADE COMMISSION Investigation Nos. 731-TA-1084-1087 (Review) Purified Carboxymethylcellulose From Finland, Mexico..., Netherlands, and Sweden: Investigation Nos. 731-TA-1084-1087(Review). By order of the Commission. Issued:...

  10. 75 FR 62100 - Purified Carboxymethylcellulose From Mexico: Final Results of Antidumping Duty Administrative Review

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-07

    ... review covers one producer/exporter, Quimica Amtex, S.A. de C.V. (Amtex). The period of review (POR) is... Carboxymethylcellulose from Mexico: Final Determination Analysis Memorandum for Quimica Amtex, S.A. de C.V.,'' dated... June 30, 2009, is as follows: Weighted- average Producer/exporter margin (percentage) Quimica Amtex,...

  11. 78 FR 11817 - Purified Carboxymethylcellulose From Finland: Final Results of Antidumping Duty Administrative...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-20

    ... analysis on December 26, 2012.\\2\\ This review covers one respondent, CP Kelco Oy and CP Kelco, Inc. (collectively CP Kelco). The period of review (POR) is July 1, 2010, through June 30, 2011. \\1\\ See Purified... respondent, CP Kelco. The petitioner in this proceeding is the Aqualon Company, a division of...

  12. 21 CFR 582.1745 - Sodium carboxymethylcellulose.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Sodium carboxymethylcellulose. 582.1745 Section... Food Additives § 582.1745 Sodium carboxymethylcellulose. (a) Product. Sodium carboxymethyl- cellulose is the sodium salt of carboxymethylcellulose not less than 99.5 percent on a dry-weight basis,...

  13. 21 CFR 582.1745 - Sodium carboxymethylcellulose.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Sodium carboxymethylcellulose. 582.1745 Section... Food Additives § 582.1745 Sodium carboxymethylcellulose. (a) Product. Sodium carboxymethyl- cellulose is the sodium salt of carboxymethylcellulose not less than 99.5 percent on a dry-weight basis,...

  14. 21 CFR 582.1745 - Sodium carboxymethylcellulose.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Sodium carboxymethylcellulose. 582.1745 Section... Food Additives § 582.1745 Sodium carboxymethylcellulose. (a) Product. Sodium carboxymethyl- cellulose is the sodium salt of carboxymethylcellulose not less than 99.5 percent on a dry-weight basis,...

  15. 21 CFR 582.1745 - Sodium carboxymethylcellulose.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Sodium carboxymethylcellulose. 582.1745 Section... Food Additives § 582.1745 Sodium carboxymethylcellulose. (a) Product. Sodium carboxymethyl- cellulose is the sodium salt of carboxymethylcellulose not less than 99.5 percent on a dry-weight basis,...

  16. 21 CFR 582.1745 - Sodium carboxymethylcellulose.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Sodium carboxymethylcellulose. 582.1745 Section... Food Additives § 582.1745 Sodium carboxymethylcellulose. (a) Product. Sodium carboxymethyl- cellulose is the sodium salt of carboxymethylcellulose not less than 99.5 percent on a dry-weight basis,...

  17. Conversion of agricultural residues to carboxymethylcellulose and carboxymethylcellulose acetate

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In view of continuing interest in the use of agricultural by-products, we have converted cellulose, wheat straw, barley straw, and rice hull into carboxymethylcellulose (CMC). Microwave-assisted synthesis was found to be a partly effective alternative to the conventional heating process. The CMC thu...

  18. 75 FR 30431 - Carboxymethylcellulose from Finland, Mexico, Netherlands, and Sweden

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-01

    ... COMMISSION Carboxymethylcellulose from Finland, Mexico, Netherlands, and Sweden AGENCY: United States... on carboxymethylcellulose from Finland, Mexico, Netherlands, and Sweden. SUMMARY: The Commission... carboxymethylcellulose from Finland, Mexico, Netherlands, and Sweden would be likely to lead to continuation...

  19. 21 CFR 182.1745 - Sodium carboxymethylcellu-lose.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Sodium carboxymethylcellu-lose. 182.1745 Section... GRAS Food Substances § 182.1745 Sodium carboxymethylcellu-lose. (a) Product. Sodium carboxy-methylcellulose is the sodium salt of carboxymethylcellulose not less than 99.5 percent on a dry-weight...

  20. 21 CFR 182.1745 - Sodium carboxymethylcellu-lose.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Sodium carboxymethylcellu-lose. 182.1745 Section... (CONTINUED) SUBSTANCES GENERALLY RECOGNIZED AS SAFE Multiple Purpose GRAS Food Substances § 182.1745 Sodium carboxymethylcellu-lose. (a) Product. Sodium carboxy-methylcellulose is the sodium salt of carboxymethylcellulose...

  1. 21 CFR 182.1745 - Sodium carboxymethylcellu-lose.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sodium carboxymethylcellu-lose. 182.1745 Section... GRAS Food Substances § 182.1745 Sodium carboxymethylcellu-lose. (a) Product. Sodium carboxy-methylcellulose is the sodium salt of carboxymethylcellulose not less than 99.5 percent on a dry-weight...

  2. 21 CFR 182.1745 - Sodium carboxymethylcellu-lose.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Sodium carboxymethylcellu-lose. 182.1745 Section... GRAS Food Substances § 182.1745 Sodium carboxymethylcellu-lose. (a) Product. Sodium carboxy-methylcellulose is the sodium salt of carboxymethylcellulose not less than 99.5 percent on a dry-weight...

  3. Purifying Nanomaterials

    NASA Technical Reports Server (NTRS)

    Hung, Ching-Cheh (Inventor); Hurst, Janet (Inventor)

    2014-01-01

    A method of purifying a nanomaterial and the resultant purified nanomaterial in which a salt, such as ferric chloride, at or near its liquid phase temperature, is used to penetrate and wet the internal surfaces of a nanomaterial to dissolve impurities that may be present, for example, from processes used in the manufacture of the nanomaterial.

  4. Water Purifier

    NASA Technical Reports Server (NTRS)

    1992-01-01

    The Floatron water purifier combines two space technologies - ionization for water purification and solar electric power generation. The water purification process involves introducing ionized minerals that kill microorganisms like algae and bacteria. The 12 inch unit floats in a pool while its solar panel collects sunlight that is converted to electricity. The resulting current energizes a specially alloyed mineral electrode below the waterline, causing release of metallic ions into the water. The electrode is the only part that needs replacing, and water purified by the system falls within EPA drinking water standards.

  5. 21 CFR 872.3500 - Polyvinylmethylether maleic anhydride (PVM-MA), acid copolymer, and carboxymethylcellulose sodium...

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...), acid copolymer, and carboxymethylcellulose sodium (NACMC) denture adhesive. 872.3500 Section 872.3500...), acid copolymer, and carboxymethylcellulose sodium (NACMC) denture adhesive. (a) Identification. Polyvinylmethylether maleic anhydride (PVM-MA), acid copolymer, and carboxymethylcellulose sodium (NACMC)...

  6. 21 CFR 872.3500 - Polyvinylmethylether maleic anhydride (PVM-MA), acid copolymer, and carboxymethylcellulose sodium...

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...), acid copolymer, and carboxymethylcellulose sodium (NACMC) denture adhesive. 872.3500 Section 872.3500...), acid copolymer, and carboxymethylcellulose sodium (NACMC) denture adhesive. (a) Identification. Polyvinylmethylether maleic anhydride (PVM-MA), acid copolymer, and carboxymethylcellulose sodium (NACMC)...

  7. 21 CFR 872.3500 - Polyvinylmethylether maleic anhydride (PVM-MA), acid copolymer, and carboxymethylcellulose sodium...

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...), acid copolymer, and carboxymethylcellulose sodium (NACMC) denture adhesive. 872.3500 Section 872.3500...), acid copolymer, and carboxymethylcellulose sodium (NACMC) denture adhesive. (a) Identification. Polyvinylmethylether maleic anhydride (PVM-MA), acid copolymer, and carboxymethylcellulose sodium (NACMC)...

  8. 21 CFR 872.3500 - Polyvinylmethylether maleic anhydride (PVM-MA), acid copolymer, and carboxymethylcellulose sodium...

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...), acid copolymer, and carboxymethylcellulose sodium (NACMC) denture adhesive. 872.3500 Section 872.3500...), acid copolymer, and carboxymethylcellulose sodium (NACMC) denture adhesive. (a) Identification. Polyvinylmethylether maleic anhydride (PVM-MA), acid copolymer, and carboxymethylcellulose sodium (NACMC)...

  9. Water Purifiers

    NASA Technical Reports Server (NTRS)

    1992-01-01

    Technology developed to purify the water aboard manned spacecraft has led to a number of spinoff applications. One of them is the Ambassador line of bacteriostatic water treatment systems, which employ high grade, high absorption media to inhibit bacteria growth and remove the medicinal taste and odor of chlorine. Company President, Ray Ward, originally became interested in the technology because of the "rusty" taste of his water supply.

  10. 21 CFR 872.3500 - Polyvinylmethylether maleic anhydride (PVM-MA), acid copolymer, and carboxymethylcellulose sodium...

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...), acid copolymer, and carboxymethylcellulose sodium (NACMC) denture adhesive. 872.3500 Section 872.3500...), acid copolymer, and carboxymethylcellulose sodium (NACMC) denture adhesive. (a) Identification... adhesive is a device composed of polyvinylmethylether maleic anhydride, acid copolymer,...

  11. 21 CFR 872.3420 - Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... polyacrylamide polymer denture adhesive. 872.3420 Section 872.3420 Food and Drugs FOOD AND DRUG ADMINISTRATION....3420 Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive. (a) Identification. A carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive is a...

  12. 21 CFR 872.3420 - Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... polyacrylamide polymer denture adhesive. 872.3420 Section 872.3420 Food and Drugs FOOD AND DRUG ADMINISTRATION....3420 Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive. (a) Identification. A carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive is a...

  13. 21 CFR 872.3420 - Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... polyacrylamide polymer denture adhesive. 872.3420 Section 872.3420 Food and Drugs FOOD AND DRUG ADMINISTRATION....3420 Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive. (a) Identification. A carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive is a...

  14. 21 CFR 872.3420 - Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... polyacrylamide polymer denture adhesive. 872.3420 Section 872.3420 Food and Drugs FOOD AND DRUG ADMINISTRATION....3420 Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive. (a) Identification. A carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive is a...

  15. 21 CFR 872.3420 - Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... polyacrylamide polymer denture adhesive. 872.3420 Section 872.3420 Food and Drugs FOOD AND DRUG ADMINISTRATION....3420 Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive. (a) Identification. A carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive is a...

  16. 75 FR 61700 - Purified Carboxymethylcellulose From Finland, the Netherlands, and Sweden: Final Results of the...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-06

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF COMMERCE... Administration, International Trade Administration, Department of Commerce. SUMMARY: On June 2, 2010, the Department of Commerce (the Department) initiated first sunset reviews of the antidumping duty orders...

  17. 76 FR 66687 - Purified Carboxymethylcellulose From the Netherlands: Final Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-27

    ..., or Angelica Mendoza, AD/CVD Operations, Office 7, Import Administration, International Trade..., International Trade Analysts, through Angelica Mendoza, Program Manager, entitled ``Analysis of Data...

  18. 75 FR 73035 - Purified Carboxymethylcellulose From Finland; Notice of Final Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-29

    ... subject to this review are CP Kelco Oy and CP Kelco U.S., Inc. (collectively, CP Kelco). The petitioner in..., ``Analysis of Data Submitted by CP Kelco Oy and CP Kelco U.S. Inc. (collectively, CP Kelco) in the... by CP Kelco Oy and CP Kelco U.S. Inc. (collectively, CP Kelco) in the 2008-2009 Administrative...

  19. 78 FR 78812 - Purified Carboxymethylcellulose From the Netherlands: Final Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-27

    ... made at less than normal value, and that CP Kelco, B.V. (CP Kelco) had no shipments of subject... noted in the Preliminary Results,\\3\\ we received a no-shipment claim from CP Kelco, and we confirmed... indicates that CP Kelco did not export subject merchandise to the United States during the POR, we...

  20. 78 FR 50028 - Purified Carboxymethylcellulose From Finland; Preliminary Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-16

    ... Hercules Inc., (Petitioner) and respondents CP Kelco Oy and CP Kelco U.S., Inc. (collectively, CP Kelco..., 2011, through June 30, 2012. We preliminarily find that CP Kelco made sales at prices below normal... section 773(b) of the Act, we disregarded certain of CP Kelco's sales in the home market that were made...

  1. 78 FR 48649 - Purified Carboxymethylcellulose From the Netherlands: Preliminary Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-09

    ... Chemicals, B.V. (Akzo Nobel) and CP Kelco, B.V. (CP Kelco). We preliminarily determine that sales of subject merchandise by Akzo Nobel were made at less than normal value and CP Kelco had no shipments of subject... received a timely submission from CP Kelco reporting to the Department that it did not sell or export...

  2. 75 FR 47788 - Purified Carboxymethylcellulose from Finland; Notice of Preliminary Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-09

    ... Hercules Inc., (the petitioner) and respondents CP Kelco Oy and CP Kelco U.S., Inc. (collectively, CP Kelco... subject merchandise to the United States produced by CP Kelco. The period of review (POR) is July 1, 2008, through June 30, 2009. We preliminarily find that CP Kelco made sales at less than normal value...

  3. 77 FR 46024 - Purified Carboxymethylcellulose From the Netherlands: Preliminary Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-02

    ... CP Kelco B.V. (CP Kelco) during the period of review of July 1, 2010, through June 30, 2011. We... during the period of review and CP Kelco had no shipments of subject merchandise during the period of... conduct an administrative review of the sales of subject merchandise from Akzo Nobel and CP Kelco...

  4. 78 FR 9884 - Purified Carboxymethylcellulose From the Netherlands: Final Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-12

    ... covers two respondents, Akzo Nobel Functional Chemicals B.V. (Akzo Nobel) and CP Kelco B.V. (CP Kelco... Preliminary Results, we received a no-shipment claim from CP Kelco, and we confirmed this claim with U.S. Customs and Border Protection (CBP). Because we find that the record indicates that CP Kelco did...

  5. 75 FR 77829 - Purified Carboxymethylcellulose From the Netherlands: Final Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-14

    ... CP Kelco B.V. (CP Kelco). The petitioner in this proceeding is Aqualon Company, a unit of Hercules... verification report for this company. We did not verify the responses of CP Kelco in the current review. On October 22, 2010, ANFC submitted a case brief and CP Kelco submitted a letter in lieu of a case brief....

  6. 77 FR 47036 - Purified Carboxymethylcellulose From Finland; Notice of Preliminary Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-07

    ... the export price (EP) or constructed export price (CEP) and NV. DATES: Effective Date: August 7, 2012...). Because the Department disregarded sales which were made at prices below the cost of production (COP) in... sales of CMC in the United States were made at less than NV, we compared U.S. price to NV, as...

  7. 75 FR 48310 - Purified Carboxymethylcellulose From the Netherlands; Preliminary Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-10

    ...-interest to Akzo Nobel Surface Chemistry B.V. and that sales of subject merchandise by ANFC and CP Kelco... treated as the successor-in- interest to Akzo Nobel Surface Chemistry B.V. (ANSC), a company for which the... Specialties (CS), which, until November 1, 2005, was part of the business unit Akzo Nobel Surface...

  8. 76 FR 36519 - Purified Carboxymethylcellulose from the Netherlands; Preliminary Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-06-22

    ... cellulose, or cellulose gum, which is a white to off-white, non-toxic, odorless, biodegradable powder..., at a minimum, reduce the remaining salt and other by-product portion of the product to less than ten... which the product was shipped directly from the Netherlands to the United States (CEP Channel 1...

  9. 75 FR 33775 - Purified Carboxymethylcellulose From Mexico: Notice of Preliminary Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-15

    ..., Department of Commerce. SUMMARY: In response to a request from Quimica Amtex S.A. de C.V. (Amtex), the... Quimica Amtex, S.A. de C.V., dated June 8, 2010 (Analysis Memorandum), for further discussion of date of... Producer/exporter margin (percentage) Quimica Amtex, S.A. de C.V 1.42 The Department will...

  10. 76 FR 20313 - Purified Carboxymethylcellulose From Mexico: Notice of Preliminary Results of Antidumping Duty...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-04-12

    ... exports of the subject merchandise to the United States produced and exported by Quimica Amtex S.A. de C.V... Quimica Amtex, S.A. de C.V., dated April 2, 2011 (Analysis Memorandum), for further discussion of date of... margin (percentage) Quimica Amtex, S.A. de C.V 0.80 The Department will disclose calculations...

  11. 76 FR 42113 - Purified Carboxymethylcellulose From Mexico: Final Results of Antidumping Duty Administrative Review

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-18

    ... one producer/ exporter, Quimica Amtex S.A. de C.V. The period of review (POR) is July 1, 2009, through...: Weighted- Producer/exporter average margin (percentage) Quimica Amtex, S.A. de C.V 0.80 Assessment...

  12. Multilayer Films and Capsules of Sodium Carboxymethylcellulose and Polyhexamethylenguanidine Hydrochloride

    NASA Astrophysics Data System (ADS)

    Guzenko, Nataliia; Gabchak, Oleksandra; Pakhlov, Evgenij

    The complexation of polyhexamethylenguanidine hydrochloride (PHMG) and sodium carboxymethylcellulose (CMC) was investigated for different conditions. Mixing of equiconcentrated aqueous solutions of the polyelectrolytes was found to result in the formation of an insoluble interpolyelectrolyte complex with an overweight of carboxymethylcellulose. A step-by-step formation of stable, irreversibly adsorbed multilayer film of the polymers was demonstrated using the quartz crystal microbalance method. Unusually thick polymer shells with a large number of loops and tails of the polyanion were formed by the method of layer-by-layer self-assembly of PHMG and CMC on spherical CaCO3 particles. Hollow multilayer capsules stable in neutral media were obtained by dissolution of the inorganic matrix in EDTA solution.

  13. Gas stream purifier

    NASA Technical Reports Server (NTRS)

    Adam, Steven J.

    1994-01-01

    A gas stream purifier has been developed that is capable of removing corrosive acid, base, solvent, organic, inorganic, and water vapors as well as particulates from an inert mixed gas stream using only solid scrubbing agents. This small, lightweight purifier has demonstrated the ability to remove contaminants from an inert gas stream with a greater than 99 percent removal efficiency. The Gas Stream Purifier has outstanding market and sales potential in manufacturing, laboratory and science industries, medical, automotive, or any commercial industry where pollution, contamination, or gas stream purification is a concern. The purifier was developed under NASA contract NAS9-18200 Schedule A for use in the international Space Station. A patent application for the Gas Stream Purifier is currently on file with the United States Patent and Trademark Office.

  14. 21 CFR 872.3410 - Ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Ethylene oxide homopolymer and/or....3410 Ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive. (a) Identification. An ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive is a...

  15. 21 CFR 872.3410 - Ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Ethylene oxide homopolymer and/or....3410 Ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive. (a) Identification. An ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive is a...

  16. 21 CFR 872.3410 - Ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Ethylene oxide homopolymer and/or....3410 Ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive. (a) Identification. An ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive is a...

  17. 21 CFR 872.3410 - Ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Ethylene oxide homopolymer and/or....3410 Ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive. (a) Identification. An ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive is a...

  18. 21 CFR 872.3410 - Ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Ethylene oxide homopolymer and/or....3410 Ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive. (a) Identification. An ethylene oxide homopolymer and/or carboxymethylcellulose sodium denture adhesive is a...

  19. METHOD FOR PURIFYING URANIUM

    DOEpatents

    Knighton, J.B.; Feder, H.M.

    1960-04-26

    A process is given for purifying a uranium-base nuclear material. The nuclear material is dissolved in zinc or a zinc-magnesium alloy and the concentration of magnesium is increased until uranium precipitates.

  20. A novel carboxymethylcellulose-gelatin-titanium dioxide-superoxide dismutase biosensor; electrochemical properties of carboxymethylcellulose-gelatin-titanium dioxide-superoxide dismutase.

    PubMed

    Emregul, Emel; Kocabay, Ozge; Derkus, Burak; Yumak, Tugrul; Emregul, Kaan Cebesoy; Sınag, Ali; Polat, Kamran

    2013-04-01

    A novel highly sensitive electrochemical carboxymethylcellulose-gelatin-TiO(2)-superoxide dismutase biosensor for the determination of O(2)(•-) was developed. The biosensor exhibits high analytical performance with a wide linear range (1.5 nM to 2 mM), low detection limit (1.5 nM), high sensitivity and low response time (1.8s). The electron transfer of superoxide dismutase was first accomplished at the carboxymethylcellulose-gelatin-Pt and carboxymethylcellulose-gelatin-TiO(2)-Pt surface. The electron transfer between superoxide dismutase and the carboxymethylcellulose-gelatin-Pt wihout Fe(CN)(6)(4-/3-) and carboxymethylcellulose-gelatin-Pt, carboxymethylcellulose-gelatin-TiO(2)-Pt with Fe(CN)(6)(4-/3-) is quasireversible with a formal potential of 200 mV, 207 mV, and 200 mV vs Ag|AgCl respectively. The anodic (ks(a)) and cathodic (ks(c)) electron transfer rate constants and the anodic (α(a)) and cathodic (α(c)) transfer coefficients were evaluated: ks(a)=6.15 s(-1), α(a)=0.79, and ks(c)=1.48 s(-1) α(c)=0.19 for carboxymethylcellulose-superoxide dismutase without Fe(CN)(6)(4-/3-), ks(a)=6.77 s(-1), α(a)=0.87, and ks(c)=1 s(-1) α(c)=0.13 for carboxymethylcellulose-superoxide dismutase with Fe(CN)(6)(4-/3-), ks(a)=6.85 s(-1), α(a)=0.88, and ks(c)=0.76 s(-1) α(c)=0.1 carboxymethylcellulose-gelatin-TiO(2)-superoxide dismutase. The electron transfer rate between superoxide dismutase and the Pt electrode is remarkably enhanced due to immobilizing superoxide dismutase in carboxymethylcellulose-gelatin and TiO(2) nanoparticles tend to act like nanoscale electrodes.

  1. Purified silicon production system

    DOEpatents

    Wang, Tihu; Ciszek, Theodore F.

    2004-03-30

    Method and apparatus for producing purified bulk silicon from highly impure metallurgical-grade silicon source material at atmospheric pressure. Method involves: (1) initially reacting iodine and metallurgical-grade silicon to create silicon tetraiodide and impurity iodide byproducts in a cold-wall reactor chamber; (2) isolating silicon tetraiodide from the impurity iodide byproducts and purifying it by distillation in a distillation chamber; and (3) transferring the purified silicon tetraiodide back to the cold-wall reactor chamber, reacting it with additional iodine and metallurgical-grade silicon to produce silicon diiodide and depositing the silicon diiodide onto a substrate within the cold-wall reactor chamber. The two chambers are at atmospheric pressure and the system is open to allow the introduction of additional source material and to remove and replace finished substrates.

  2. Method of purifying isosaccharinate

    DOEpatents

    Rai, Dhanpat; Moore, Robert C.; Tucker, Mark D.

    2010-09-07

    A method of purifying isosaccharinate by mixing sodium carbonate, potassium carbonate, sodium hydroxide or potassium hydroxide with calcium isosaccharinate, removing the precipitated calcium carbonate and adjusting the pH to between approximately 4.5 to 5.0 thereby removing excess carbonate and hydroxide to provide an acidic solution containing isosaccharinate.

  3. Purifying Water by Imbibition

    NASA Technical Reports Server (NTRS)

    Lawton, E. A.

    1986-01-01

    Concept for purifying water uses absorbent material to remove organic substances. Entire bulk of material employed, not just surface. Proposed purification process uses inexpensive equipment and low energy. Material is methyl acrylate polymer. Material cheap and regenerated by rinsing with methanol or by allowing absorbed compounds to evaporate from it.

  4. Carboxymethylcellulose film for bacterial wound infection control and healing.

    PubMed

    Wong, Tin Wui; Ramli, Nor Amlizan

    2014-11-04

    Infection control and wound healing profiles of sodium carboxymethylcellulose (SCMC) films were investigated as a function of their anti-bacterial action, physical structures, polymer molecular weights and carboxymethyl substitution degrees. The films were prepared with in vitro polymer/film and in vivo microbe-colonized wound healing/systemic infection profiles examined. Adhesive high carboxymethyl substituted SCMC films aided healing via attaching to microbes and removing them from wound. Pseudomonas aeruginosa was removed via encapsulating in gelling low molecular weight SCMC film, whereas Staphylococcus aureus was trapped in tight folds of high molecular weight SCMC film. Incomplete microbe removal from wound did not necessary translate to inability to heal as microbe remnant at wound induced fibroblast migration and aided tissue reconstruction. Using no film nonetheless will cause systemic blood infection. SCMC films negate infection and promote wound healing via specific polymer-microbe adhesion, and removal of S. aureus and P. aeruginosa requires films of different polymer characteristics.

  5. PROCESS OF PURIFYING URANIUM

    DOEpatents

    Seaborg, G.T.; Orlemann, E.F.; Jensen, L.H.

    1958-12-23

    A method of obtaining substantially pure uranium from a uranium composition contaminated with light element impurities such as sodium, magnesium, beryllium, and the like is described. An acidic aqueous solution containing tetravalent uranium is treated with a soluble molybdate to form insoluble uranous molybdate which is removed. This material after washing is dissolved in concentrated nitric acid to obtaln a uranyl nitrate solution from which highly purified uranium is obtained by extraction with ether.

  6. 75 FR 60084 - Purified Carboxymethylcellulose from Mexico: Preliminary Results of the First Five-year (“Sunset...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-29

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF COMMERCE..., International Trade Administration, Department of Commerce. SUMMARY: On June 2, 2010, the Department of Commerce... conducting a full sunset review of the antidumping duty order on CMC from Mexico, pursuant to section...

  7. 76 FR 4865 - Purified Carboxymethylcellulose From Mexico: Final Results of the First Five-Year (“Sunset...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-27

    ..., polyanionic cellulose, or cellulose gum, which is a white to off-white, non-toxic, odorless, biodegradable... operations, which, at a minimum, reduce the remaining salt and other by-product portion of the product...

  8. 76 FR 38609 - Antidumping or Countervailing Duty Order, Finding, or Suspended Investigation; Opportunity To...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-01

    ...: Carboxymethylcellulose, A-421-811....... 7/1/10-6/30/11 Russia: Solid Urea, A-821-801 7/1/10-6/30/11 Ferrovanadium and... Republic of China: Carbon Steel Butt-Weld Pipe Fittings, A-570-814.. 7/1/10-6/30/11 Certain Potassium...- 1/1/10-12/31/10 825 Italy: Certain Pasta, C-475-819 1/1/10-12/31/10 The People's Republic of...

  9. Natural Air Purifier

    NASA Technical Reports Server (NTRS)

    1993-01-01

    NASA environmental research has led to a plant-based air filtering system. Dr. B.C. Wolverton, a former NASA engineer who developed a biological filtering system for space life support, served as a consultant to Terra Firma Environmental. The company is marketing the BioFilter, a natural air purifier that combines activated carbon and other filter media with living plants and microorganisms. The filter material traps and holds indoor pollutants; plant roots and microorganisms then convert the pollutants into food for the plant. Most non-flowering house plants will work. After pollutants have been removed, the cleansed air is returned to the room through slits in the planter. Terra Firma is currently developing a filter that will also disinfect the air.

  10. Synthesis and Characterization of Carboxymethylcellulose-Methacrylate Hydrogel Cell Scaffolds

    PubMed Central

    Reeves, Robert; Ribeiro, Andreia; Lombardo, Leonard; Boyer, Richard; Leach, Jennie B.

    2012-01-01

    Many carbohydrates pose advantages for tissue engineering applications due to their hydrophilicity, degradability, and availability of chemical groups for modification. For example, carboxymethylcellulose (CMC) is a water-soluble cellulose derivative that is degradable by cellulase. Though this enzyme is not synthesized by mammalian cells, cellulase and the fragments derived from CMC degradation are biocompatible. With this in mind, we created biocompatible, selectively degradable CMC-based hydrogels that are stable in routine culture, but degrade when exposed to exogenous cellulase. Solutions of CMC-methacrylate and polyethylene glycol dimethacrylate (PEG-DM) were co-crosslinked to form stable hydrogels; we found that greater CMC-methacrylate content resulted in increased gel swelling, protein diffusion and rates of degradation by cellulase, as well as decreased gel shear modulus. CMC-methacrylate/PEG-DM gels modified with the adhesive peptide RGD supported fibroblast adhesion and viability. We conclude that hydrogels based on CMC-methacrylate are suitable for bioengineering applications where selective degradability may be favorable, such as cell scaffolds or controlled release devices. PMID:22708058

  11. Synthesis of cellulose acetate and carboxymethylcellulose from sugarcane straw.

    PubMed

    Candido, R G; Gonçalves, A R

    2016-11-05

    Sugarcane straw (SCS) is a raw material with high potential for production of cellulose derivatives due to its morphology and structure. The proposal of this work was to synthesize cellulose acetate (CA) and carboxymethylcellulose (CMC) from sugarcane straw cellulose, and applied the CA in the preparation of a membrane. The cellulose extraction was carried out in four steps. Firstly, SCS was treated with H2SO4 (10% v/v) followed by NaOH (5% w/v) treatment. Subsequently, a chelating process was performed before ending the extraction process with chemical bleaching using H2O2 (5% v/v). The extracted cellulose was employed in the obtainment of CA and CMC. The CA presented a degree of substitution (DS) of 2.72. Its FTIR spectrum showed that practically all hydroxyl groups were replaced by acetate groups. The membrane synthesized from CA was dense and homogeneous. The presence of small particles on the top and bottom surfaces decreased the mechanical resistance of the membrane. The CMC presented a low DS (0.4) demonstrating the carboxymethylation reaction was not very effective due to the presence of lignin. These results proved that SCS can be utilized in the synthesis of CA and CMC.

  12. Injectable carboxymethylcellulose hydrogels for soft tissue filler applications.

    PubMed

    Varma, Devika M; Gold, Gittel T; Taub, Peter J; Nicoll, Steven B

    2014-12-01

    Disease, trauma and aging all lead to deficits in soft tissue. As a result, there is a need to develop materials that safely and effectively restore areas of deficiency. While autogenous fat is the current gold standard, hyaluronic acid (HA) fillers are commonly used. However, the animal and bacterial origin of HA-based materials can induce adverse reactions in patients. With the aim of developing a safer and more affordable alternative, this study characterized the properties of a plant-derived, injectable carboxymethylcellulose (CMC) soft tissue filler. Specifically, methacrylated CMC was synthesized and crosslinked to form stable hydrogels at varying macromer concentrations (2-4% w/v) using an ammonium persulfate and ascorbic acid redox initiation system. The equilibrium Young's modulus was shown to vary with macromer concentration (ranging from ∼2 to 9.25kPa), comparable to values of native soft tissue and current surgical fillers. The swelling properties were similarly affected by macromer concentration, with 4% gels exhibiting the lowest swelling ratio and mesh size, and highest crosslinking density. Rheological analysis was performed to determine gelation onset and completion, and was measured to be within the ISO standard for injectable materials. In addition, hydrolytic degradation of these gels was sensitive to macromer concentration, while selective removal using enzymatic treatment was also demonstrated. Moreover, favorable cytocompatibility of the CMC hydrogels was exhibited by co-culture with human dermal fibroblasts. Taken together, these findings demonstrate the tunability of redox-crosslinked CMC hydrogels by varying fabrication parameters, making them a versatile platform for soft tissue filler applications.

  13. Development of biodegradable aluminium carboxymethylcellulose matrices for mosquito larvicides.

    PubMed

    Mathew, Nisha; Kalyanasundaram, Muthuswami

    2004-07-01

    The use of ecofriendly biodegradable controlled-release formulations of mosquito larvicides could reduce the frequency of application and losses due to degradation of the insecticide compared with conventional formulations. Among the 20 matrices developed by entrapping the organophosphorus mosquito larvicide, fenthion, in carboxymethylcellulose ionotropically cross-linked with aluminium ions which were studied for release profiles, two matrices, CRF3b and CRF5b, were found to be stable for 16 and 14 weeks under simulated field conditions. The average concentration of fenthion released per week ranged from 0.06 to 3.5 mg litre(-1) for CRF3b and 0.09 to 2.72 mg litre(-1) for CRF5b. Of these two formulations, CRF3b was the more stable, maintaining the concentration of the active ingredient at the level required to effect mosquito control. The cumulative release of fenthion per pellet was 80% from CRF3b and 72% from CRF5b. Based on the study with fenthion, two similar matrices for triflumuron, a benzoylphenylurea insect growth regulator, STAR3b and STAR5b were developed. These matrices were stable up to 16 weeks with the average concentration of triflumuron released per week ranging from 0.05 to 3.44 mg litre(-1) for STAR3b and 0.07 to 2.71 mg litre(-1) for STARSb. The cumulative release of triflumuron per pellet was 75% from STAR3b and 76% from STAR5b. From the results of this study under simulated conditions, it is estimated that the application of four pellets of either fenthion or triflumuron per square metre of the breeding surface may play a useful role in controlling Culex quinquefasciatus Say in larval habitats for about 4 months.

  14. Tranexamic Acid and Hyaluronate/Carboxymethylcellulose Create Cell Injury

    PubMed Central

    Yılmaz, Bayram; Dilbaz, Serdar; Üstün, Yusuf; Kumru, Selahattin

    2014-01-01

    Background and Objectives: Postoperative pelvic adhesions are associated with chronic pelvic pain, dyspareunia, and infertility. The aim of this study was to evaluate the adhesion prevention effects of tranexamic acid (TA) and hyaluronate/carboxymethylcellulose (HA/CMC) barrier in the rat uterine horn models on the basis of macroscopic and microscopic adhesion scores and histopathological as well as biochemical parameters of inflammation. Methods: Twenty-one Wistar rats were randomly divided into 3 groups. Ten lesions were created on the antimesenteric surface of both uterine horns by bipolar cautery. Three milliliters of 0.9% sodium chloride solution were administered in the control group. A single layer of 2 × 2 cm HA/CMC was plated in group 2. Two milliliters of TA was applied in the last group. All rats were sacrificed at postoperative day 21. Results: No significant difference was found among the control group, the HA/CMC group, and the TA group in terms of macro-adhesion score (P = .206) and microadhesion score (P = .056). No significant difference was found among the 3 groups in terms of inflammation score (P = .815) and inflammatory cell activity (P = .835). Malondialdehyde levels were significantly lower in the control group than in the TA group and HA/CMC group (P = .028). Superoxide dismutase and glutathione S-transferase activities were found to be higher in the control group than in the TA group (P = .005) and HA/CMC group (P = .009). Conclusions: TA and HA/CMC had no efficacy in preventing macroscopic or microscopic adhesion formation and decreasing inflammatory cell activity or inflammation score in our rat models. TA and HA/CMC increased the levels of free radicals and reduced the activities of superoxide dismutase and glutathione S-transferase enzymes, which act to reduce tissue injury. PMID:25392658

  15. Eco-friendly Synthesis of Ceria Foam via Carboxymethylcellulose Gelation: Application for the Epoxidation of Chalcone

    EPA Science Inventory

    A simple and innovative process is described for the eco-friendly preparation of ceria foams via the carboxymethylcellulose gelation by Ce4+ cations; heat treatment of the ensuing xerogels produces ceria foams. The influence of the concentration of cerium and of the calcination t...

  16. Polyion-counterion interactions in sodium carboxymethylcellulose-ethylene glycol-water ternary solutions.

    PubMed

    Sharma, Ramesh; Das, Chanchal; Dahal, Sanjay; Das, Bijan

    2013-02-15

    Polyion-counterion interactions in sodium carboxymethylcellulose-ethylene glycol-water ternary system have been investigated with special reference to their variations with the polyelectrolyte concentration, the medium relative permittivity and the temperature using electrical conductance measurements. The experimental data have been analyzed on the basis of a model for semidilute polyelectrolyte conductivity which takes into account the scaling arguments proposed by Dobrynin et al. Concentration-dependent moderate counterion condensation (24-33%) was observed. Counterion condensation is found to be spontaneous which depends upon the EG-content of the medium and the temperature. A major proportion of the current is transported by the carboxymethylcellulose polyions. The results further demonstrated that the monomer units experience more frictional resistance in solution as the EG-content increases or as the temperature decreases.

  17. Methods for purifying carbon materials

    DOEpatents

    Dailly, Anne; Ahn, Channing; Yazami, Rachid; Fultz, Brent T.

    2009-05-26

    Methods of purifying samples are provided that are capable of removing carbonaceous and noncarbonaceous impurities from a sample containing a carbon material having a selected structure. Purification methods are provided for removing residual metal catalyst particles enclosed in multilayer carbonaceous impurities in samples generate by catalytic synthesis methods. Purification methods are provided wherein carbonaceous impurities in a sample are at least partially exfoliated, thereby facilitating subsequent removal of carbonaceous and noncarbonaceous impurities from the sample. Methods of purifying carbon nanotube-containing samples are provided wherein an intercalant is added to the sample and subsequently reacted with an exfoliation initiator to achieve exfoliation of carbonaceous impurities.

  18. Purifying Aluminum by Vacuum Distillation

    NASA Technical Reports Server (NTRS)

    Du Fresne, E. R.

    1985-01-01

    Proposed method for purifying aluminum employs one-step vacuum distillation. Raw material for process impure aluminum produced in electrolysis of aluminum ore. Impure metal melted in vacuum. Since aluminum has much higher vapor pressure than other constituents, boils off and condenses on nearby cold surfaces in proportions much greater than those of other constituents.

  19. Process for purifying geothermal steam

    DOEpatents

    Li, Charles T.

    1980-01-01

    Steam containing hydrogen sulfide is purified and sulfur recovered by passing the steam through a reactor packed with activated carbon in the presence of a stoichiometric amount of oxygen which oxidizes the hydrogen sulfide to elemental sulfur which is adsorbed on the bed. The carbon can be recycled after the sulfur has been recovered by vacuum distillation, inert gas entrainment or solvent extraction. The process is suitable for the purification of steam from geothermal sources which may also contain other noncondensable gases.

  20. Process for purifying geothermal steam

    DOEpatents

    Li, C.T.

    Steam containing hydrogen sulfide is purified and sulfur recovered by passing the steam through a reactor packed with activated carbon in the presence of a stoichiometric amount of oxygen which oxidizes the hydrogen sulfide to elemental sulfur which is adsorbed on the bed. The carbon can be recycled after the sulfur has been recovered by vacuum distillation, inert gas entrainment or solvent extraction. The process is suitable for the purification of steam from geothermal sources which may also contain other noncondensable gases.

  1. Vaginal inserts based on chitosan and carboxymethylcellulose complexes for local delivery of chlorhexidine: preparation, characterization and antimicrobial activity.

    PubMed

    Bigucci, Federica; Abruzzo, Angela; Vitali, Beatrice; Saladini, Bruno; Cerchiara, Teresa; Gallucci, Maria Caterina; Luppi, Barbara

    2015-01-30

    The aim of this work was to prepare vaginal inserts based on chitosan/carboxymethylcellulose polyelectrolyte complexes for local delivery of chlorhexidine digluconate. Complexes were prepared with different chitosan/carboxymethylcellulose molar ratios at a pH value close to pKa interval of the polymers and were characterized in terms of physico-chemical properties, complexation yield and drug loading. Then complexes were used to prepare inserts as vaginal dosage forms and their physical handling, morphology, water-uptake ability and drug release properties as well as antimicrobial activity toward Candida albicans and Escherichia coli were evaluated. Results confirmed the ionic interaction between chitosan and carboxymethylcellulose and the influence of the charge amount on the complexation yield. Complexes were characterized by high values of drug loading and showed increasing water-uptake ability with the increase of carboxymethylcellulose amount. The selection of appropriate chitosan/carboxymethylcellulose molar ratios allowed to obtain cone-like shaped solid inserts, easy to handle and able to hydrate releasing the drug over time. Finally, the formulated inserts showed antimicrobial activity against common pathogens responsible for vaginal infections.

  2. Poor fluorinated graphene sheets carboxymethylcellulose polymer composite mode locker for erbium doped fiber laser

    NASA Astrophysics Data System (ADS)

    Mou, Chengbo; Arif, Raz; Lobach, Anatoly S.; Khudyakov, Dmitry V.; Spitsina, Nataliya G.; Kazakov, Valery A.; Turitsyn, Sergei; Rozhin, Aleksey

    2015-02-01

    We report poor fluorinated graphene sheets produced by thermal exfoliation embedding in carboxymethylcellulose polymer composite (GCMC) as an efficient mode locker for erbium doped fiber laser. Two GCMC mode lockers with different concentration have been fabricated. The GCMC based mode locked fiber laser shows stable soliton output pulse shaping with repetition rate of 28.5 MHz and output power of 5.5 mW was achieved with the high concentration GCMC, while a slightly higher output power of 6.9 mW was obtained using the low concentration GCMC mode locker.

  3. Synthesis of carboxymethylcellulose/acrylic acid hydrogels with superabsorbent properties by radiation-initiated crosslinking

    NASA Astrophysics Data System (ADS)

    Fekete, Tamás; Borsa, Judit; Takács, Erzsébet; Wojnárovits, László

    2016-07-01

    Superabsorbent hydrogels were prepared by gamma irradiation from aqueous solutions of carboxymethylcellulose (CMC) and acrylic acid (AAc) with varying CMC:AAc ratio. By partially replacing the CMC with AAc the gelation increased and led to a higher gel fraction and lower water uptake. Moreover, the gelation required significantly milder synthesis conditions. Decreasing both the dose and the solute concentration in the presence of AAc led to gels with higher gel fraction and higher degree of swelling compared to pure CMC gels. Increasing the AAc content up to 10% proved to be very effective, while very high AAc content (over 50%) hindered the gelation process.

  4. Poor fluorinated graphene sheets carboxymethylcellulose polymer composite mode locker for erbium doped fiber laser

    SciTech Connect

    Mou, Chengbo E-mail: a.rozhin@aston.ac.uk; Turitsyn, Sergei; Rozhin, Aleksey E-mail: a.rozhin@aston.ac.uk; Arif, Raz; Lobach, Anatoly S.; Spitsina, Nataliya G.; Khudyakov, Dmitry V.; Kazakov, Valery A.

    2015-02-09

    We report poor fluorinated graphene sheets produced by thermal exfoliation embedding in carboxymethylcellulose polymer composite (GCMC) as an efficient mode locker for erbium doped fiber laser. Two GCMC mode lockers with different concentration have been fabricated. The GCMC based mode locked fiber laser shows stable soliton output pulse shaping with repetition rate of 28.5 MHz and output power of 5.5 mW was achieved with the high concentration GCMC, while a slightly higher output power of 6.9 mW was obtained using the low concentration GCMC mode locker.

  5. Process for purifying zirconium sponge

    SciTech Connect

    Abodishish, H.A.M.; Kimball, L.S.

    1992-03-31

    This patent describes a Kroll reduction process wherein a zirconium sponge contaminated with unreacted magnesium and by-product magnesium chloride is produced as a regulus, a process for purifying the zirconium sponge. It comprises: distilling magnesium and magnesium chloride from: a regulus containing a zirconium sponge and magnesium and magnesium chloride at a temperature above about 800{degrees} C and at an absolute pressure less than about 10 mmHg in a distillation vessel to purify the zirconium sponge; condensing the magnesium and the magnesium chloride distilled from the zirconium sponge in a condenser; and then backfilling the vessel containing the zirconium sponge and the condenser containing the magnesium and the magnesium chloride with a gas; recirculating the gas between the vessel and the condenser to cool the zirconium sponge from above about 800{degrees} C to below about 300{degrees} C; and cooling the recirculating gas in the condenser containing the condensed magnesium and the condensed magnesium chloride as the gas cools the zirconium sponge to below about 300{degrees} C.

  6. Methods for Purifying Enzymes for Mycoremediation

    NASA Technical Reports Server (NTRS)

    Cullings, Kenneth W. (Inventor); DeSimone, Julia C. (Inventor); Paavola, Chad D. (Inventor)

    2014-01-01

    A process for purifying laccase from an ectomycorrhizal fruiting body is disclosed. The process includes steps of homogenization, sonication, centrifugation, filtration, affinity chromatography, ion exchange chromatography, and gel filtration. Purified laccase can also be separated into isomers.

  7. Formulation of Novel Layered Sodium Carboxymethylcellulose Film Wound Dressings with Ibuprofen for Alleviating Wound Pain

    PubMed Central

    Vinklárková, Lenka; Vetchý, David; Bernatonienė, Jurga

    2015-01-01

    Effective assessment and management of wound pain can facilitate both improvements in healing rates and overall quality of life. From a pharmacological perspective, topical application of nonsteroidal anti-inflammatory drugs in the form of film wound dressings may be a good choice. Thus, the aim of this work was to develop novel layered film wound dressings containing ibuprofen based on partially substituted fibrous sodium carboxymethylcellulose (nonwoven textile Hcel NaT). To this end, an innovative solvent casting method using a sequential coating technique has been applied. The concentration of ibuprofen which was incorporated as an acetone solution or as a suspension in a sodium carboxymethylcellulose dispersion was 0.5 mg/cm2 and 1.0 mg/cm2 of film. Results showed that developed films had adequate mechanical and swelling properties and an advantageous acidic surface pH for wound application. An in vitro drug release study implied that layered films retained the drug for a longer period of time and thus could minimize the frequency of changing the dressing. Films with suspended ibuprofen demonstrated higher drug content uniformity and superior in vitro drug release characteristics in comparison with ibuprofen incorporation as an acetone solution. Prepared films could be potential wound dressings for the effective treatment of wound pain in low exuding wounds. PMID:26090454

  8. 21 CFR 872.3490 - Carboxymethylcellulose sodium and/or polyvinylmethylether maleic acid calcium-sodium double salt...

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... polyvinylmethylether maleic acid calcium-sodium double salt denture adhesive. 872.3490 Section 872.3490 Food and Drugs... maleic acid calcium-sodium double salt denture adhesive. (a) Identification. A carboxymethylcellulose sodium and/or polyvinylmethylether maleic acid calcium-sodium double salt denture adhesive is a...

  9. 21 CFR 872.3490 - Carboxymethylcellulose sodium and/or polyvinylmethylether maleic acid calcium-sodium double salt...

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... polyvinylmethylether maleic acid calcium-sodium double salt denture adhesive. 872.3490 Section 872.3490 Food and Drugs... maleic acid calcium-sodium double salt denture adhesive. (a) Identification. A carboxymethylcellulose sodium and/or polyvinylmethylether maleic acid calcium-sodium double salt denture adhesive is a...

  10. 21 CFR 872.3490 - Carboxymethylcellulose sodium and/or polyvinylmethylether maleic acid calcium-sodium double salt...

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... polyvinylmethylether maleic acid calcium-sodium double salt denture adhesive. 872.3490 Section 872.3490 Food and Drugs... maleic acid calcium-sodium double salt denture adhesive. (a) Identification. A carboxymethylcellulose sodium and/or polyvinylmethylether maleic acid calcium-sodium double salt denture adhesive is a...

  11. 21 CFR 872.3490 - Carboxymethylcellulose sodium and/or polyvinylmethylether maleic acid calcium-sodium double salt...

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... polyvinylmethylether maleic acid calcium-sodium double salt denture adhesive. 872.3490 Section 872.3490 Food and Drugs... maleic acid calcium-sodium double salt denture adhesive. (a) Identification. A carboxymethylcellulose sodium and/or polyvinylmethylether maleic acid calcium-sodium double salt denture adhesive is a...

  12. 21 CFR 872.3490 - Carboxymethylcellulose sodium and/or polyvinylmethylether maleic acid calcium-sodium double salt...

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... polyvinylmethylether maleic acid calcium-sodium double salt denture adhesive. 872.3490 Section 872.3490 Food and Drugs... maleic acid calcium-sodium double salt denture adhesive. (a) Identification. A carboxymethylcellulose sodium and/or polyvinylmethylether maleic acid calcium-sodium double salt denture adhesive is a...

  13. Turbocharged engine with exhaust purifier

    SciTech Connect

    Tadokoro, T.; Matsuda, I.; Okimoto, H.

    1986-09-23

    The patent described a control system for an automobile engine having intake and exhaust systems for respectively conducting intake gases to and exhaust gases from the engine, which comprises, in combination: a turbocharger including a turbine disposed in the exhaust system and adapted to be driven by the flow of the exhaust gases therethrough and a blower disposed in the intake system and drivingly connected with the turbine for supercharging the intake gases; and exhaust purifying device disposed in the exhaust system downstream of the turbine with respect to the direction of flow of the exhaust gases; a regulating means for varying the effective cross-section of a portion of the exhaust system leading to the turbine; a control means for controlling the regulating means in dependence on an operating condition of the engine, the control means causing the regulating means to decrease the effective cross-section during a low speed operating condition, but to increase the effective cross-section during a high speed operating condition of the engine.

  14. Microstructure of Purified Rubber Particles.

    PubMed

    Wood; Cornish

    2000-05-01

    Purified rubber particles from Hevea brasiliensis (Brazilian rubber tree), Parthenium argentatum (guayule), Ficus elastica (Indian rubber tree), and Euphorbia lactiflua were examined and compared using conventional scanning electron microscopy (SEM), field-emission SEM, cryo-SEM, and transmission electron microscopy (TEM). Rubber particles of all four species were spherical; they varied in size and had a uniform homogeneous material, the rubber core, surrounded by a contiguous monolayer (half-unit) membrane. Frozen-hydrated and/or untreated particles from H. brasiliensis and P. argentatum deformed and fused readily, whereas those from F. elastica and E. lactiflua retained their spherical shapes. These results indicate that the surface components of the H. brasiliensis and P. argentatum particles are more fluid than those of F. elastica or E. lactiflua. When fixed in aldehyde, F. elastica particles retained their spherical exterior shapes but had hollow centers, whereas H. brasiliensis and P. argentatum particles completely collapsed. In aldehyde-osmium tetroxide-fixed material, the rubber core of F. elastica was poorly preserved in some particles in which only a small amount of the rubber core remained adhering to the monolayer membrane, leaving a hollow center. Euphorbia lactiflua particles were well preserved in terms of retaining the rubber core; however, the membrane was not as easily discernible as it was in the other three species. Both H. brasiliensis and P. argentatum were well preserved following fixation; their cores remained filled with rubber, and their monolayer membranes were defined. The addition of potassium permanganate to the fixation-staining regime resulted in higher-contrast micrographs and more well defined monolayer membranes.

  15. Hydrothermal synthesis of magnetic mesoporous carbon microspheres from carboxymethylcellulose and nickel acetate

    NASA Astrophysics Data System (ADS)

    Wu, Qiong; Li, Wei; Tan, Jia; Nan, Xi; Liu, Shouxin

    2015-03-01

    Paramagnetic mesoporous carbon spheres with diameters of 1-3 μm were synthesized through the hydrothermal carbonization of carboxymethylcellulose with nickel acetate, followed by high-temperature carbonization in a N2 atmosphere. Monodisperse Ni particles of average size of 2-5 nm were doped into the carbon matrix, and covered the entrances of pores. Ni particles existed as metallic nickel and nickel oxide with ordered lattice structures. The effect of Ni content on the specific surface area, mesopore percentage, and magnetic and adsorption properties were investigated. The highest vitamin B12 adsorption capacity of 103 mg/g was achieved for the sample prepared using 0.04 g of nickel acetate. The Freundlich and Langmuir isotherm models were used to determine the equilibrium uptakes of vitamin B12. Vitamin B12 was physically adsorbed as a monolayer on the carbon spheres. The carbon spheres were easily separated on account of their magnetism.

  16. Application of carboxymethylcellulose hydrogel based silver nanocomposites on cotton fabrics for antibacterial property.

    PubMed

    Bozaci, Ebru; Akar, Emine; Ozdogan, Esen; Demir, Asli; Altinisik, Aylin; Seki, Yoldas

    2015-12-10

    In this study, fumaric acid (FA) crosslinked carboxymethylcellulose (CMC) hydrogel (CMCF) based silver nanocomposites were coated on cotton fabric for antibacterial property for the first time. The performance of the nanocomposite treated cotton fabric was tested for different mixing times of hydrogel solution, padding times and concentrations of silver. The cotton fabrics treated with CMC hydrogel based silver nanocomposites demonstrated 99.9% reduction for both Staphylococcus aureus (Sa) and Klebsiella pneumonia (Kp). After one cycle washing processes of treated cotton fabric, there is no significant variation observed in antibacterial activity. From SEM and AFM analyses, silver particles in nano-size, homogenously distributed, were observed. The treated samples were also evaluated by tensile strength, thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FTIR) analysis, fluid absorbency properties, and whiteness index. The treatment of cotton fabric with CMCF hydrogel did not affect the whiteness considerably, but increased the absorbency values of cotton.

  17. Impact of carboxymethylcellulose on morphology and antibiotic production by Streptomyces hygroscopicus.

    PubMed

    Ilić, Slavica B; Konstantinović, Sandra S; Veljković, Vlada B; Savić, Dragisa S; Lazić, Miodrag L; Gojgić-Cvijović, Gordana

    2008-07-01

    A chemically defined media consisting of carboxymethylcellulose (CMC) was developed to maximize the production of antibiotics, hexaene H-85 and azalomycine, by Streptomyces hygroscopicus CH-7. The production of antibiotics by filamentous organisms is often dependent on the morphology and size distribution of the pellet population within the culture. By adding the polymer to the fermentation medium, the growth was changed from a single large glob to small reproducible pellets, and wall growth was diminished to a minimum. Maximum concentrations of hexaene H-85 (146.7 mg/dm(3)) and azalomycine (188.6 mg/dm(3)) were reached at 3.0% and 1.0% (w/v) CMC, respectively.

  18. Effects of carboxymethylcellulose and carboxypolymethylene on morphology of Aspergillus fumigatus NRRL 2346 and fumagillin production.

    PubMed

    Yang, Wen; Hartwieg, Erika A; Fang, Aiqi; Demain, Arnold L

    2003-01-01

    Aspergillus fumigatus NRRL 2346 is the producer of fumagillin, an antitumor antibiotic that inhibits angiogenesis. This strain is very difficult to grow reproducibly in shake flasks owing to an extreme form of pellet growth and extensive wall growth. The effects of carboxymethylcellulose (CMC) and carboxypolymethylene (Carbopol) on growth and fumagillin production by A. fumigatus were investigated. By adding the polymers to the fermentation medium, the growth form of the mold was changed from a single large glob to small reproducible pellets, and wall growth was diminished to a minimum. Carbopol, at a lower concentration, was more effective than CMC in improving both morphology and production. Small pellets were produced which favored fumagillin biosynthesis. 1.5% (wt/vol) CMC and 0.3% (wt/vol) Carbopol were found to be the optimum concentrations; higher levels increased viscosity to an unacceptable level.

  19. Hydrogen purifier module with membrane support

    DOEpatents

    A hydrogen purifier utilizing a hydrogen-permeable membrane to purify hydrogen from mixed gases containing hydrogen is disclosed. Improved mechanical support for the permeable membrane is described, enabling forward or reverse differential pressurization of the membrane, which further stabilizes the membrane from wrinkling upon hydrogen uptake.

    2012-07-24

    A hydrogen purifier utilizing a hydrogen-permeable membrane to purify hydrogen from mixed gases containing hydrogen is disclosed. Improved mechanical support for the permeable membrane is described, enabling forward or reverse differential pressurization of the membrane, which further stabilizes the membrane from wrinkling upon hydrogen uptake.

  20. Method for purifying bidentate organophosphorus compounds

    DOEpatents

    Schulz, Wallace W.

    1977-01-01

    Bidentate organophosphorus compounds useful for extracting actinide elements from acidic nuclear waste solutions are purified of undesirable acidic impurities by contacting the compounds with ethylene glycol which preferentially extracts the impurities found in technical grade bidentate compounds.

  1. Carboxymethylcellulose-gelatin-superoxidase dismutase electrode for amperometric superoxide radical sensing.

    PubMed

    Kocabay, Ozge; Emregul, Emel; Aras, Sümer; Emregul, Kaan Cebesoy

    2012-08-01

    A novel, highly sensitive superoxide dismutase biosensor for the direct and simultaneous determination of superoxide radicals was developed by immobilization of superoxide dismutase within carboxymethylcellulose-gelatin on a Pt electrode surface. The parameters affecting the performance of the biosensor were investigated. The response of the CMC-G-SOD biosensor was proportional to O (2) (·-) concentration and the detection limit was 1.25 × 10(-3) mM with a correlation coefficient of 0.9994. The developed biosensor exhibited high analytical performance with wider linear range, high sensitivity and low response time. The biosensor retained 89.8% of its sensitivity after use for 80 days. The support system enhanced the immobilization of superoxide dismutase and promoted the electron transfer of superoxide dismutase minimizing its fouling effect. The biosensor was quite effective not only in detecting O (2) (·-) , but also in determining the antioxidant properties of acetylsalicylic acid-based drugs and the anti-radical activity of healthy and cancerous human brain tissues.

  2. Novel biocomposite of carboxymethyl chitosan and pineapple peel carboxymethylcellulose as sunscreen carrier.

    PubMed

    Wongkom, Lucksanee; Jimtaisong, Ampa

    2017-02-01

    This study aims to prepare of biocomposite of carboxymethyl chitosan (CM-chitosan) and carboxymethylcellulose (CMC) from Ananas comosus (pineapple) peel for use as broad spectrum sunscreen carrier. Biocomposite was produced by using ferulic acid (FA), a plant extract, as crosslinker with the optimal ratio of CMC: CM-chitosan: FA at 1:2:4%w. FT-IR technique demonstrated that crosslinking may occur at amine group of CM-chitosan and carboxyl group of FA and hydrogen bonding between hydroxyl group of CMC and carboxyl group of FA. Biocomposite is pale yellow powder and present fibre bundle-like surface in the SEM image. DSC, TGA and XRD results indicated that new compound was formed. The particle size of biocomposite is 626nm determined by using Zetasizer. Hydrophilic TiO2 and phenylbenzimidazole sulphonic acid (PBSA) were used as sunscreen agent at ratio of TiO2: PBSA at 2:1%w. The biocomposite sunscreen possesses the SPF value of 2.47 with boost star rating of 3 at 2% compound. The results obtained indicate that the biocomposite was successfully prepared from CM-chitosan and pineapple peel CMC and the system can be used as matrix delivery system for hydrophilic sunscreens.

  3. Association and Diffusion of Li(+) in Carboxymethylcellulose Solutions for Environmentally Friendly Li-ion Batteries.

    PubMed

    Casalegno, Mosè; Castiglione, Franca; Passarello, Marco; Mele, Andrea; Passerini, Stefano; Raos, Guido

    2016-07-21

    Carboxymethylcellulose (CMC) has been proposed as a polymeric binder for electrodes in environmentally friendly Li-ion batteries. Its physical properties and interaction with Li(+) ions in water are interesting not only from the point of view of electrode preparation-processability in water is one of the main reasons for its environmental friendliness-but also for its possible application in aqueous Li-ion batteries. We combine molecular dynamics simulations and variable-time pulsed field gradient spin-echo (PFGSE) NMR spectroscopy to investigate Li(+) transport in CMC-based solutions. Both the simulations and experimental results show that, at concentrations at which Li-CMC has a gel-like consistency, the Li(+) diffusion coefficient is still very close to that in water. These Li(+) ions interact preferentially with the carboxylate groups of CMC, giving rise to a rich variety of coordination patterns. However, the diffusion of Li(+) in these systems is essentially unrestricted, with a fast, nanosecond-scale exchange of the ions between CMC and the aqueous environment.

  4. Preparation and characterization of silver nanoparticle loaded amorphous hydrogel of carboxymethylcellulose for infected wounds.

    PubMed

    Das, Anup; Kumar, Ajay; Patil, Niranjan B; Viswanathan, Chandra; Ghosh, Deepa

    2015-10-05

    There is a growing demand for an appropriate and safe antimicrobial dressing to treat infected deep wounds. An amorphous gel formulation (SNP-CMC), containing silver nanoparticles (SNPs) and carboxymethylcellulose (CMC), was prepared in one step by the reduction of silver nitrate in situ. Spectrophotometric and microscopic analysis revealed that the SNPs were 7-21 nm in diameter. In simulated wound experiments, SNP-CMC gel was found to absorb 80.48 ± 4.69% w/w of saline and donate 17.43 ± 0.76% w/w of moisture within 24h indicating its dual fluid affinity. Cytocompatibility of the gel was assessed by proliferation studies with primary human skin cells. The antimicrobial activity studies showed that SNP-CMC containing 50 ppm of SNPs was effective against the growth of both Gram negative and Gram positive strains including methicillin-resistant Staphylococcus aureus (MRSA). These results indicate that SNP-CMC could be ideal for the treatment of deep infected wounds.

  5. Very fast dissolving acid carboxymethylcellulose-rifampicin matrix: Development and solid-state characterization.

    PubMed

    Luciani-Giacobbe, Laura C; Ramírez-Rigo, María V; Garro-Linck, Yamila; Monti, Gustavo A; Manzo, Ruben H; Olivera, María E

    2017-01-01

    One of the main obstacles to the successful treatment of tuberculosis is the poor and variable oral bioavailability of rifampicin (RIF), which is mainly due to its low hydrophilicity and dissolution rate. The aim of this work was to obtain a hydrophilic new material that allows a very fast dissolution rate of RIF and therefore is potentially useful in the development of oral solid dosage forms. The acid form of carboxymethylcellulose (CMC) was co-processed with RIF by solvent impregnation to obtain CMC-RIF powder, which was characterized by polarized optical microscopy, powder x-ray diffraction, DSC-TGA, hot stage microscopy, (13)C and (15)N solid-state NMR and FT-IR spectroscopy. In addition, the CMC-RIF matrices were subjected to water uptake and dissolution studies to assess hydrophilicity and release kinetics. CMC-RIF is a crystalline solid dispersion. Solid-state characterization indicated that no ionic interaction occurred between the components, but RIF crystallized as a zwitterion over the surface of CMC, which drastically increased the hydrophilicity of the solid. The CMC-RIF matrices significantly improved the water uptake of RIF and disintegrated in a very short period immediately releasing RIF. As CMC improves the hydrophilicity and delivery properties of RIF, CMC-RIF is very useful in the design of oral solid dosage forms with very fast dissolution of RIF, either alone or in combination with other antitubercular drugs.

  6. Effects of carboxymethylcellulose and hydroxypropylmethylcellulose on the differentiation and activity of osteoclasts and osteoblasts.

    PubMed

    Agis, Hermann; Beirer, Benedict; Watzek, Georg; Gruber, Reinhard

    2010-11-01

    Carboxymethylcellulose (CMC) and hydroxypropylmethylcellulose (HPMC) serve as carriers for growth factors and bone substitutes. Although both carriers are placed into the defect sites, their impacts on bone regeneration are unclear. Herein, we examined whether CMC and HPMC affect the differentiation of bone marrow progenitors into osteoclasts and osteoblasts. We therefore induced osteoclastogenesis and osteoblastogenesis in murine bone marrow progenitors in the presence of CMC and HPMC, respectively. Measures of osteoclastogesis were based on the number and activity of tartrate-resistant acid-phosphatase-positive (TRAP(+)) multinucleated cells and expression of marker genes. Osteoblastogenesis was determined by the number and activity of alkaline-phosphatase-positive (AP(+)) colonies and relevant marker genes. Viability was assessed by colorimetric measurement of formazan formation. We report that CMC at 1% caused a significant reduction in the number and activity of TRAP(+) multinucleated cells. Changes in viability were not responsible for the observed effects. HPMC showed no remarkable impact on osteoclastogenesis; however, the concentration was limited to 0.5% because of the high viscosity. The ability of bone marrow progenitors to form AP(+) colonies was not affected by either of the two carriers. Together, these results suggest that CMC and possibly also HPMC can decrease osteoclastogenesis while osteoblastogenesis remains unchanged in vitro. These observations raise the possibility that these carriers might affect the cellular process of bone regeneration.

  7. Efficacy of Carboxymethylcellulose and Hyaluronate in Dry Eye Disease: A Systematic Review and Meta-Analysis

    PubMed Central

    Song, Jae Kyeong; Park, Hwa Yeon; Hyon, Joon Young; Oh, Seung-Won; Bae, Woo Kyung; Han, Jong-Soo; Jung, Se Young; Um, Yoo Jin; Lee, Ga-Hye; Yang, Ji Hye

    2017-01-01

    Background The efficacy of two artificial tears, carboxymethylcellulose (CMC) and hyaluronate (HA), was compared in the treatment of patients with dry eye disease. Methods We conducted a systematic review and meta-analysis on randomized controlled trials in the PubMed, Embase, Cochrane Library, and ClinicalTrials.gov databases. The efficacy was compared in terms of the mean change from baseline in tear break-up time. The meta-analysis was conducted using both random and fixed effect models. The quality of the selected studies was assessed for risk of bias. Results Five studies were included involving 251 participants. Random effect model meta-analysis showed no significant difference between CMC and HA in treating dry eye disease (pooled standardized mean difference [SMD]=-0.452; 95% confidence interval [CI], -0.911 to 0.007; P=0.053). In contrast, fixed effect model meta-analysis revealed significant improvements in the CMC group when compared to the HA group (pooled SMD=-0.334; 95% CI, -0.588 to -0.081; P=0.010). Conclusion The efficacy of CMC appeared to be better than that of HA in treating dry eye disease, although meta-analysis results were not statistically significant. Further research is needed to better elucidate the difference in efficacy between CMC and HA in treating dry eye disease. PMID:28197326

  8. Field-scale transport and transformation of carboxymethylcellulose-stabilized nano zero-valent iron.

    PubMed

    Johnson, Richard L; Nurmi, James T; O'Brien Johnson, Graham S; Fan, Dimin; O'Brien Johnson, Reid L; Shi, Zhenqing; Salter-Blanc, Alexandra J; Tratnyek, Paul G; Lowry, Gregory V

    2013-02-05

    The fate of nano zerovalent iron (nZVI) during subsurface injection was examined using carboxymethylcellulose (CMC) stabilized nZVI in a very large three-dimensional physical model aquifer with detailed monitoring using multiple, complementary detection methods. A fluorescein tracer test in the aquifer plus laboratory column data suggested that the very-aggressive flow conditions necessary to achieve 2.5 m of nZVI transport could be obtained using a hydraulically constrained flow path between injection and extraction wells. However, total unoxidized nZVI was transported only about 1 m and <2% of the injected nZVI concentration reached that distance. The experimental data also indicated that groundwater flow changed during injection, likely due to hydrogen bubble formation, which diverted the nZVI away from the targeted flow path. The leading edge of the iron plume became fully oxidized during transport. However, within the plume, oxidation of nZVI decreased in a fashion consistent with progressive depletion of aquifer "reductant demand". To directly quantify the extent of nZVI transport, a spectrophotometric method was developed, and the results indicated that deployment of unoxidized nZVI for groundwater remediation will likely be difficult.

  9. Carboxymethylcellulose Obtained by Ethanol/Water Organosolv Process Under Acid Conditions

    NASA Astrophysics Data System (ADS)

    Ruzene, Denise S.; Gonçalves, Adilson R.; Teixeira, José A.; Pessoa de Amorim, Maria T.

    Sugar cane bagasse pulps were obtained by ethanol/water organosolv process under acid and alkaline conditions. The best condition of acid pulping for the sugarcane bagasse was 0.02 mol/L sulfuric acid at 160°C, for 1h, whereas the best condition for alkaline pulping was 5% sodium hydroxide (base pulp) at 160°C, for 3h. For the residual lignin removal, the acid and alkaline pulps were submitted to a chemical bleaching using sodium chlorite. Pulps under acid and alkaline conditions bleached with sodium chlorite presented viscosities of 3.6 and 7.8 mPas, respectively, and μ-kappa numbers of 1.1 and 2.4, respectively. The pulp under acid condition, bleached with sodium chlorite was used to obtain carboxymethylcellulose (CMC). CMC yield was 35% (pulp based), showing mass gain after the carboxymethylation reaction corresponding to 23.6% of substitution or 0.70 groups-CH2COONa per unit of glucose residue. The infrared spectra showed the CMC characteristic bands and by the infrared technique it was possible to obtain a substitution degree (0.63), similar to the substitution degree calculated by mass gain (0.70).

  10. Preparation and characterization of glycerol plasticized-pea starch/ZnO-carboxymethylcellulose sodium nanocomposites.

    PubMed

    Yu, Jiugao; Yang, Jingwen; Liu, Baoxiang; Ma, Xiaofei

    2009-06-01

    Among natural polymers, starch is one of the most promising biodegradable materials because it is a renewable bioresource that is universally available and of low cost. However, the properties of starch-based materials are not satisfactory. One approach is the use of nano-filler as reinforcement for starch-based materials. In this paper, a nanocomposite is prepared using ZnO nanoparticles stabilized by carboxymethylcellulose sodium (CMC) as the filler in glycerol plasticized-pea starch (GPS) matrix by the casting process. According to the characterization of ZnO-CMC particles with Fourier transform infrared (FTIR), Ultraviolet-visible (UV-vis), X-ray diffraction (XRD), transmission electron microscope (TEM) and thermogravimetric analysis (TG), ZnO (about 60 wt%) is encapsulated with CMC (about 40 wt%) in ZnO-CMC particles with the size of about 30-40 nm. A low loading of ZnO-CMC particles can obviously improve the pasting viscosity, storage modulus, the glass transition temperature and UV absorbance of GPS/ZnO-CMC nanocomposites. When the ZnO-CMC contents vary from 0 to 5 wt%, the tensile yield strength increase from 3.94 MPa to 9.81 MPa, while the elongation at break reduce from 42.2% to 25.8%. The water vapor permeability decrease from 4.76 x 10(-10) to 1.65 x 10(-10) g m(-1) s(-1) Pa(-1).

  11. Method for purifying bidentate organophosphorous compounds

    DOEpatents

    McIsaac, Lyle D.; Krupa, Joseph F.; Schroeder, Norman C.

    1981-01-01

    Bidentate organophosphorous compounds are purified of undesirable impurities by contacting a solution of the compounds with a mercuric nitrate solution to form an insoluble mercuric bidentate compound which precipitates while the impurities remain in solution. The precipitate is washed and then contacted with a mixture of an aqueous solution of a strong mercuric ion complexing agent and an organic solvent to complex the mercuric ion away from the bidentate compound which then dissolves in the solvent. The purified bidentate compounds are useful for extracting the actinide elements from aqueous acidic nuclear waste solutions.

  12. Lidocaine carboxymethylcellulose with gelatine co-polymer hydrogel delivery by combined microneedle and ultrasound.

    PubMed

    Nayak, Atul; Babla, Hiten; Han, Tao; Das, Diganta Bhusan

    2016-01-01

    A study that combines microneedles (MNs) and sonophoresis pre-treatment was explored to determine their combined effects on percutaneous delivery of lidocaine from a polymeric hydrogel formulation. Varying ratios of carboxymethylcellulose and gelatine (NaCMC/gel ranges 1:1.60-1:2.66) loaded with lidocaine were prepared and characterized for zeta potential and particle size. Additionally, variations in the formulation drying techniques were explored during the formulation stage. Ex vivo permeation studies using Franz diffusion cells measured lidocaine permeation through porcine skin after pre-treatment with stainless steel MNs and 20 kHz sonophoresis for 5-and 10-min durations. A stable formulation was related to a lower gelatine mass ratio because of smaller mean particle sizes and high zeta potential. Lidocaine permeability in skin revealed some increases in permeability from combined MN and ultrasound pre-treatment studies. Furthermore, up to 4.8-fold increase in the combined application was observed compared with separate pre-treatments after 30 min. Sonophoresis pre-treatment alone showed insignificant enhancement in lidocaine permeation during the initial 2 h period. MN application increased permeability at a time of 0.5 h for up to ∼17 fold with an average up to 4 fold. The time required to reach therapeutic levels of lidocaine was decreased to less than 7 min. Overall, the attempted approach promises to be a viable alternative to conventional lidocaine delivery methods involving painful injections by hypodermic needles. The mass transfer effects were fairly enhanced and the lowest amount of lidocaine in skin was 99.7% of the delivered amount at a time of 3 h for lidocaine NaCMC/GEL 1:2.66 after low-frequency sonophoresis and MN treatment.

  13. Synthesis and structure-activity study of quaternary ammonium functionalized beta-cyclodextrin-carboxymethylcellulose polymers.

    PubMed

    Bonenfant, Danielle; Bourgeois, François-René; Mimeault, Murielle; Monette, Frédéric; Niquette, Patrick; Hausler, Robert

    2011-01-01

    Carboxymethylcellulose (CMC) and beta-cyclodextrin (beta-CD)-based polymers functionalized with two types of quaternary ammonium compounds (QACs), the alkaquat DMB-451 (N-alkyl (50% C14, 40% C12, 10% C10) dimethylbenzylammonium chloride) (DMD-451) named polymer DMB-451, and FMB 1210-8 (a blend of 32 w% N-alkyl (50% C14, 40% C12, 10% C10) dimethylbenzylammonium chloride and 48 w% of didecyldimethylammonium chloride) named polymer FMB 1210-8, were synthethized and characterized by Fourier transform infrared spectroscopy. The antimicrobial activities of these polymers against Eschericia coli were also evaluated at 25 degrees C in wastewater. The results have indicated that the polymer FMB 1210-8 possesses a high-affinity binding with bacterial cells that induces a rapid disinfection process. Moreover, in the same experimental conditions of disinfection (mixture of 1.0 g of polymer and 100 mL of wastewater), the polymer FMB 1210-8 has a higher antimicrobial efficiency (99.90%) than polymer DMB-451 (92.8%). This phenomenon might be associated to a stronger interaction with bacterial cells due to stronger binding affinity for E. coli cells and greater killing efficiency of the C10 alkyl chains QAC of polymer FMB 1210-8 to disrupt the bacterial cell membrane as compared to N-alkyl (50% C14, 40% C12, 10% C10) dimethylbenzylammonium chloride. Together, these results suggest that the polymer FMB 1210-8 could constitute a good disinfectant against Escherichia coli, which could be advantageously used in wastewater treatments due to the low toxicity of beta-CD and CMC, and moderated toxicity of FMB 1210-8 to human and environment.

  14. Direct synthesis of magnetite nanoparticles from iron(II) carboxymethylcellulose and their performance as NMR contrast agents

    NASA Astrophysics Data System (ADS)

    da Silva, Delmarcio Gomes; Hiroshi Toma, Sergio; de Melo, Fernando Menegatti; Carvalho, Larissa Vieira C.; Magalhães, Alvicler; Sabadini, Edvaldo; dos Santos, Antônio Domingues; Araki, Koiti; Toma, e. Henrique E.

    2016-01-01

    Iron(II) carboxymethylcellulose (CMC) has been successfully employed in the synthesis of hydrophylic magnetite nanoparticles stabilized with a biopolymer coating, aiming applications in NMR imaging. The new method encompasses a convenient one-step synthetic procedure, allowing a good size control and yielding particles of about 10 nm (core size). In addition to the biocompatibility, the nanoparticles have promoted a drastic reduction in the transverse relaxation time (T2) of the water protons. The relaxivity rates have been investigated as a function of the nanoparticles concentration, showing a better performance in relation to the common NMR contrast agents available in the market.

  15. Dermatitis from purified sea algae toxin (debromoaplysiatoxin).

    PubMed

    Solomon, A E; Stoughton, R B

    1978-09-01

    Cutaneous inflammation was induced by debromoaplysiatoxin, a purified toxin extracted from Lyngbya majuscula Gomont. This alga causes a seaweed dermatitis that occurs in persons who have swum off the coast of Oahu in Hawaii. By topical application, the toxin was found to produce an irritant pustular folliculitis in humans and to cause a severe cutaneous inflammatory reaction in the rabbit and in hairless mice.

  16. Improvement of Linde Kryotechnik's internal purifier

    NASA Astrophysics Data System (ADS)

    Decker, Lutz; Meier, Albert; Wilhelm, Hanspeter

    2014-01-01

    With the recent shortage in supply of helium, recovery solutions have experienced a new focus with a tendency to recover streams with higher impurity content. This development calls for purifier systems operating efficiently and with low impact on liquefaction capacity for helium streams with impurity levels in the percentage range. Linde Kryotechnik has answered this demand by improving the performance of its purifier technology. Since 1983, its standardized helium liquefiers of the L- and former TCF-series type contain an internal purifier which already allows efficient impurity removal with minimized space demand. Along with a line dryer to absorb humidity, it is designed to remove air impurities up to 5 mol%. However, with increasing impurity level, liquefaction capacity reduced significantly being furthermore restricted to an upper level of approx. 180 l/h and continuous purification became limited in time. With the current redesign of this purifier, the impact on liquefaction capacity is now minimized without any limitation within the capacity range of the L-series plants. Continuous purification is hence ensured beyond previous maximum impurity content. This paper provides the key design changes and the achievable performance, which has been verified in the recent L-series plants delivered to customers.

  17. Method of purifying neutral organophosphorus extractants

    DOEpatents

    Horwitz, E. Philip; Gatrone, Ralph C.; Chiarizia, Renato

    1988-01-01

    A method for removing acidic contaminants from neutral mono and bifunctional organophosphorous extractants by contacting the extractant with a macroporous cation exchange resin in the H.sup.+ state followed by contact with a macroporous anion exchange resin in the OH.sup.- state, whereupon the resins take up the acidic contaminants from the extractant, purifying the extractant and improving its extraction capability.

  18. Home Air Purifiers Eradicate Harmful Pathogens

    NASA Technical Reports Server (NTRS)

    2014-01-01

    Marshall Space Flight Center funded the University of Madison-Wisconsin to develop ethylene scrubbers to keep produce fresh in space. Akida Holdings of Jacksonville, Florida, licensed the technology and developed Airocide, an air purifier that can kill airborne pathogens. Previously designed for industrial spaces, there is now a specially designed unit for home use.

  19. Two systems developed for purifying inert atmospheres

    NASA Technical Reports Server (NTRS)

    Foster, M. S.; Johnson, C. E.; Kyle, M. L.

    1969-01-01

    Two systems, one for helium and one for argon, are used for purifying inert atmospheres. The helium system uses an activated charcoal bed at liquid nitrogen temperature to remove oxygen and nitrogen. The argon system uses heated titanium sponge to remove nitrogen and copper wool beds to remove oxygen. Both use molecular sieves to remove water vapor.

  20. Electrophoretic separator for purifying biologicals, part 1

    NASA Technical Reports Server (NTRS)

    Mccreight, L. R.

    1978-01-01

    A program to develop an engineering model of an electrophoretic separator for purifying biologicals is summarized. An extensive mathematical modeling study and numerous ground based tests were included. Focus was placed on developing an actual electrophoretic separator of the continuous flow type, configured and suitable for flight testing as a space processing applications rocket payload.

  1. Effect of sodium carboxymethylcellulose and fucidic acid on the gel characterization of polyvinylalcohol-based wound dressing.

    PubMed

    Lim, Soo-Jeong; Lee, Jeong Hoon; Piao, Ming Guan; Lee, Mi-Kyung; Oh, Dong Hoon; Hwang, Du Hyung; Quan, Qi Zhe; Yong, Chul Soon; Choi, Han-Gon

    2010-07-01

    The purpose of this study was to investigate the effect of sodium carboxymethylcellulose (Na-CMC) and fucidic acid on the gel characterization for the development of sodium fucidate-loaded wound dressing. The cross-linked hydrogel films were prepared with polyvinyl alcohol (PVA) and sodium carboxymethylcellulose (Na-CMC) using the freeze-thawing method. Their gel properties such as gel fraction, swelling, water vapor transmission test, morphology, tensile strength and thermal property were investigated. In vitro protein adsorption test and release were performed. Na-CMC decreased the gel fraction and tensile strength of the hydrogels, but increased the swelling ability, water vapor transmission rate, elasticity and porosity of hydrogels. Thus, the wound dressing developed with PVA and Na-CMC was more swellable, flexible and elastic than that with only PVA because of its cross-linking interaction with PVA. However, the drug had a negative effect on the gel properties of hydrogels but there were no significant differences. In particular, the hydrogel composed of 2.5% PVA, 1.125% Na-CMC and 0.2% drug might give an adequate level of moisture and build up the exudates on the wound area. Thus, this sodium fucidate-loaded hydrogel could be a potential candidate for wound dressing with excellent forming.

  2. Effects of cellulose, carboxymethylcellulose and inulin fed to rats as single supplements or in combinations on their caecal parameters.

    PubMed

    Juśkiewicz, Jerzy; Zduńczyk, Zenon

    2004-12-01

    We compared the effect of diets containing different nondigestible carbohydrates: cellulose (C), inulin (IN) and carboxymethylcellulose (CMC) as single supplements or in dietary combination on caecal physiology of rats. Sixty male Wistar rats (Rattus norvegicus) were divided into five groups and for 4 weeks were fed a casein diet with the compared carbohydrates (4% of diet) or a combination of IN+C or IN+CMC (both 4+4%). Diet intake and FCR index remained unaffected by the treatments, whereas IN improved the body weight gain of rats compared to CMC. Compared to C group, all diets containing IN and CMC decreased the caecal pH as well as enlarged the caecum, thus increasing the weights of contents and tissue, especially upon CMC treatment. Rats given carboxymethylcellulose (CMC and IN+CMC groups) had watery caecal digesta, and some of them suffered from diarrhoea. In the case of CMC, the caecal enlargement was due to tissue hypertrophy and digesta accumulation mostly in response to an increased bulk of contents. Unlike C+IN, the dietary combination of CMC- and inulin-enhanced fermentation in the caecum of rats, however the proportion of acetate, propionate and butyrate was less beneficial. Compared to CMC, inulin gave a higher concentration of SCFA, especially of butyrate and propionate. The action of inulin in the caecum of rats could be pronounced by dietary treatment combined with CMC.

  3. Steroidogenesis in amlodipine treated purified Leydig cells

    SciTech Connect

    Latif, Rabia; Lodhi, Ghulam Mustafa; Hameed, Waqas; Aslam, Muhammad

    2012-01-01

    Drugs have been shown to adversely affect male fertility and recently anti-hypertensive drugs were added to the list. The anti-fertility effects of amlodipine, a calcium channel blocker, are well-illustrated in in vivo experiments but lack an in vitro proof. The present study was designed to experimentally elucidate the effects of amlodipine on Leydig cell steroidogenesis and intracellular calcium in vitro. Leydig cells of Sprague–Dawley rats were isolated and purified by Percoll. Cells were incubated for 3 h with/without amlodipine in the presence/absence of LH, dbcAMP, Pregnenolone and 25-Hydroxycholesterol. Cytosolic calcium was measured in purified Leydig cells by fluorometric technique. The results showed significantly reduced (P < 0.05) steroidogenesis and intracellular calcium in amlodipine exposed rats. The site of amlodipine induced steroidogenic inhibition seems to be prior to the formation of Pregnenolone at the level of StAR protein. -- Highlights: ► Inhibition of steroidogenesis in isolated and purified Leydig cells by amlodipine. ► Site of inhibition was before Pregnenolone formation, at the level of StAR protein. ► Inhibition of LH stimulated rise in cytosolic calcium by amlodipine.

  4. Utilization of purified cellulose in fiber studies.

    PubMed

    Penner, M H; Liaw, E T

    1990-01-01

    Purified cellulose-type fiber products are widely used in experimental nutrition. Their use in a broad spectrum of studies may potentially lead to the acceptance of the misconception that the various commercially available cellulose products are equivalent. In this paper we have attempted to show that this is not the case. The comparative structural data of Table 2 and the compositional data of Olsen et al provide examples which indicate that purified cellulose preparations should not necessarily be considered equivalent. Unfortunately, our current lack of understanding of how fibers are metabolized and how they may affect specific physiological parameters makes it difficult to determine which, if any, of the measurable structural and chemical properties will be of relevance for a given in vivo study. At present, it appears that researchers utilizing/evaluating the consequences of consuming a purified cellulose-type fiber would be prudent to provide at least a limited amount of data on the properties of the cellulose preparation used in their studies. The characterization of the cellulose product may be done by a variety of methods depending on the expertise of the laboratory. The methods and results discussed in this paper provide an example of the type of information which may be obtained from an in vitro characterization of cellulose products.

  5. Air Purifiers Eliminate Pathogens, Preserve Food

    NASA Technical Reports Server (NTRS)

    2009-01-01

    NASA-funded researchers produced an ethylene reduction device for a plant growth unit. KES Science & Technology Inc., a Kennesaw, Georgia-based company specializing in sustaining perishable foods, licensed the ethylene scrubbing technology. KES partnered with Akida Holdings, of Jacksonville, Florida, which now markets the NASA-developed technology as AiroCide. According to the company, it is the only air purifier that completely destroys airborne bacteria, mold, fungi, mycotoxins, viruses, volatile organic compounds (like ethylene), and odors. What?s more, the devices have no filters that need changing and produce no harmful byproducts, such as the ozone created by some filtration systems.

  6. Interactions and compatibility of 11 S globulin from Vicia faba seeds and sodium salt of carboxymethylcellulose in an aqueous medium.

    PubMed

    Antonov, Yu A; Dmitrochenko, A P; Leontiev, A L

    2006-02-28

    This work studies specific interactions and compatibility between a legumin and a linear carboxylated polysaccharide using gel permeation chromatography, sedimentation analysis, SDS gel electrophoresis, viscometry and phase analysis measurements. It uses the system water/11 S globulin/CMC as a model. Carboxymethylcellulose (CMC) molecules are able to cause a partial dissociation of the protein, subsequent formation of soluble interbiopolymeric complexes and partial aggregation of the free non combined protein at room temperature and pH 6.0-6.5. The maximal binding of biopolymers is observed at their equimolar ratio. The decrease in temperature of the mixture from 293 to 277 K leads to formation of the complex coacervate. The increase in pH from 6.0 to 7.6 results in suppression of complex formation and manifestation of the phenomenon of thermodynamic incompatibility if the total concentration of biopolymers in the system exceeds the critical concentration of segregative phase separation.

  7. Radiation crosslinking of carboxymethylcellulose of various degree of substitution at high concentration in aqueous solutions of natural pH

    NASA Astrophysics Data System (ADS)

    Wach, Radoslaw A.; Mitomo, Hiroshi; Nagasawa, Naotsugu; Yoshii, Fumio

    2003-12-01

    Carboxymethylcellulose (CMC) hydrogel formed by ionizing radiation at highly concentrated aqueous solutions was found to undergo swelling depending on the pH of the swelling media. Swelling increases at neutral and basic pH due to ionization of carboxymethyl groups on side chains. The presence of charges develops repulsive forces between polymer chains of the network causing its expansion. Hydrogel in relaxed state as well as dried gel reveals good mechanical properties. It was considered that intermolecular crosslinking reactions occur by a radical route. Radicals placed on anhydroglucose repeating unit as well as on side chains were distinguished from ESR spectra of CMC. A stable doublet signal with 2.0 mT splitting constant belongs to a radical placed on the α-carbon atom of the substituent group, R-O- rad CH-COO -. It was assumed that this species participates in intermolecular crosslinking.

  8. Carboxymethylcellulose adsorption on molybdenite: the effect of electrolyte composition on adsorption, bubble-surface collisions, and flotation.

    PubMed

    Kor, Mohammad; Korczyk, Piotr M; Addai-Mensah, Jonas; Krasowska, Marta; Beattie, David A

    2014-10-14

    The adsorption of carboxymethylcellulose polymers on molybdenite was studied using spectroscopic ellipsometry and atomic force microscopy imaging with two polymers of differing degrees of carboxyl group substitution and at three different electrolyte conditions: 1 × 10(-2) M KCl, 2.76 × 10(-2) M KCl, and simulated flotation process water of multicomponent electrolyte content, with an ionic strength close to 2.76 × 10(-2) M. A higher degree of carboxyl substitution in the adsorbing polymer resulted in adsorbed layers that were thinner and with more patchy coverage; increasing the ionic strength of the electrolyte resulted in increased polymer layer thickness and coverage. The use of simulated process water resulted in the largest layer thickness and coverage for both polymers. The effect of the adsorbed polymer layer on bubble-particle attachment was studied with single bubble-surface collision experiments recorded with high-speed video capture and image processing and also with single mineral molybdenite flotation tests. The carboxymethylcellulose polymer with a lower degree of substitution resulted in almost complete prevention of wetting film rupture at the molybdenite surface under all electrolyte conditions. The polymer with a higher degree of substitution prevented rupture only when adsorbed from simulated process water. Molecular kinetic theory was used to quantify the effect of the polymer on the dewetting dynamics for collisions that resulted in wetting film rupture. Flotation experiments confirmed that adsorbed polymer layer properties, through their effect on the dynamics of bubble-particle attachment, are critical to predicting the effectiveness of polymers used to prevent mineral recovery in flotation.

  9. 21 CFR 880.6500 - Medical ultraviolet air purifier.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Miscellaneous Devices § 880.6500 Medical ultraviolet air purifier. (a) Identification. A medical ultraviolet air purifier is a device intended for medical purposes that is used to destroy bacteria in the air by...

  10. 21 CFR 880.6500 - Medical ultraviolet air purifier.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... Miscellaneous Devices § 880.6500 Medical ultraviolet air purifier. (a) Identification. A medical ultraviolet air purifier is a device intended for medical purposes that is used to destroy bacteria in the air by...

  11. 21 CFR 880.6500 - Medical ultraviolet air purifier.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... Miscellaneous Devices § 880.6500 Medical ultraviolet air purifier. (a) Identification. A medical ultraviolet air purifier is a device intended for medical purposes that is used to destroy bacteria in the air by...

  12. 21 CFR 880.6500 - Medical ultraviolet air purifier.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... Miscellaneous Devices § 880.6500 Medical ultraviolet air purifier. (a) Identification. A medical ultraviolet air purifier is a device intended for medical purposes that is used to destroy bacteria in the air by...

  13. Apparatus and methods for purifying lead

    DOEpatents

    Tunison, Harmon M.

    2016-01-12

    Disclosed is an exemplary method of purifying lead which includes the steps of placing lead and a fluoride salt blend in a container; forming a first fluid of molten lead at a first temperature; forming a second fluid of the molten fluoride salt blend at a second temperature higher than the first temperature; mixing the first fluid and the second fluid together; separating the two fluids; solidifying the molten fluoride salt blend at a temperature above a melting point of the lead; and removing the molten lead from the container. In certain exemplary methods the molten lead is removed from the container by decanting. In still other exemplary methods the molten salt blend is a Lewis base fluoride eutectic salt blend, and in yet other exemplary methods the molten salt blend contains sodium fluoride, lithium fluoride, and potassium fluoride.

  14. Induction slag reduction process for purifying metals

    DOEpatents

    Traut, Davis E.; Fisher, II, George T.; Hansen, Dennis A.

    1991-01-01

    A continuous method is provided for purifying and recovering transition metals such as neodymium and zirconium that become reactive at temperatures above about 500.degree. C. that comprises the steps of contacting the metal ore with an appropriate fluorinating agent such as an alkaline earth metal fluosilicate to form a fluometallic compound, and reducing the fluometallic compound with a suitable alkaline earth or alkali metal compound under molten conditions, such as provided in an induction slag metal furnace. The method of the invention is advantageous in that it is simpler and less expensive than methods used previously to recover pure metals, and it may be employed with a wide range of transition metals that were reactive with enclosures used in the prior art methods and were hard to obtain in uncontaminated form.

  15. Method of separating and purifying gadolinium-153

    DOEpatents

    Bray, Lane A [Richland, WA; Corneillie, Todd M [Davis, CA

    2001-01-01

    The present invention is an improvement to the method of separating and purifying gadolinium from a mixture of gadolinium and europium having the steps of (a) dissolving the mixture in an acid; (b) reducing europium+3 to europium+2; and (c) precipitating the europium+2 with a sulfate ion in a superstoichiometric amount; wherein the improvement is achieved by using one or more of the following: (i) the acid is an anoic acid; (ii) the reducing is with zinc metal in the absence of a second metal or with an amount of the second metal that is ineffective in the reducing; (iii) adding a group IIA element after step (c) for precipitating the excess sulfate prior to repeating step (c); (iv) the sulfate is a sulfate salt with a monovalent cation; (v) adding cold europium+3 prior to repeating step (c).

  16. Some enzymatic activities associated with purified parapoxvirions.

    PubMed Central

    Caplen, H S; Holowczak, J A

    1983-01-01

    Purified virions of milker's nodule virus, a parapoxvirus, were shown to contain an RNA polymerase, a nucleotide phosphohydrolase, and a protein kinase associated with or encapsulated within the DNA-containing core of the virus. In vitro, the activated viral RNA polymerase transcribed only 7 to 8% of the genome, in the form of 8S to 14S polyadenylated RNA molecules which were complementary to sequences present in milker's nodule virus DNA but not vaccinia virus DNA or DNA prepared from the host cells in which the virus was propagated. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis showed that in vitro, the activated viral protein kinase phosphorylated viral polypeptides of 95, 60, 33.5, 15, and 13.8 kilodaltons. Images PMID:6188861

  17. Ozone emissions from a "personal air purifier".

    PubMed

    Phillips, T J; Bloudoff, D P; Jenkins, P L; Stroud, K R

    1999-01-01

    Ozone emissions were measured above a "personal air purifier" (PAP) designed to be worn on a lapel, shirt pocket, or neck strap. The device is being marketed as a negative ion generator that purifies the air. However, it also produces ozone within the person's immediate breathing zone. In order to assess worst-case potential human exposure to ozone at the mouth and nose, we measured ozone concentrations in separate tests at 1, 3, 5, and 6 in. above each of two PAPs in a closed office. One PAP was new, and one had been used slightly for 3 months. Temperature, relative humidity, atmospheric pressure, room ozone concentration, and outdoor ozone concentration also were measured concurrently during the tests. Average ozone levels measured directly above the individual PAPs ranged from 65-71 ppb at 6 in. above the device to 268-389 ppb at 1 in. above the device. Ozone emission rates from the PAPs were estimated to be 1.7-1.9 microg/minute. When house dust was sprinkled on the top grid of the PAPs, one showed an initial peak of 522 ppb ozone at 1 in., and then returned to the 200-400 ppb range. Room ozone levels increased by only 0-5 ppb during the tests. Even when two PAPs were left operating over a weekend, room ozone levels did not noticeably increase beyond background room ozone levels. These results indicate that this "PAP," even without significant background ozone, can potentially elevate the user's exposures to ozone levels greater than the health-based air quality standards for outdoor air in California (0.09 ppm, 1-hour average) and the United States (0.08 ppm, 8-hour average).

  18. 21 CFR 880.6710 - Medical ultraviolet water purifier.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Medical ultraviolet water purifier. 880.6710... Miscellaneous Devices § 880.6710 Medical ultraviolet water purifier. (a) Identification. A medical ultraviolet water purifier is a device intended for medical purposes that is used to destroy bacteria in water...

  19. 21 CFR 880.6710 - Medical ultraviolet water purifier.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Medical ultraviolet water purifier. 880.6710... Miscellaneous Devices § 880.6710 Medical ultraviolet water purifier. (a) Identification. A medical ultraviolet water purifier is a device intended for medical purposes that is used to destroy bacteria in water...

  20. 21 CFR 880.6710 - Medical ultraviolet water purifier.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Medical ultraviolet water purifier. 880.6710... Miscellaneous Devices § 880.6710 Medical ultraviolet water purifier. (a) Identification. A medical ultraviolet water purifier is a device intended for medical purposes that is used to destroy bacteria in water...

  1. 21 CFR 880.6710 - Medical ultraviolet water purifier.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Medical ultraviolet water purifier. 880.6710... Miscellaneous Devices § 880.6710 Medical ultraviolet water purifier. (a) Identification. A medical ultraviolet water purifier is a device intended for medical purposes that is used to destroy bacteria in water...

  2. Pasting properties of Tamarind (Tamarindus indica) kernel powder in the presence of Xanthan, Carboxymethylcellulose and Locust bean gum in comparison to Rice and Potato flour.

    PubMed

    Kaur, Maninder; Sandhu, Kawaljit Singh; Kaur, Jasmeen

    2013-08-01

    Effects of addition of different levels of gums (xanthan, carboxymethylcellulose and locust bean gum) on the pasting properties of tamarind kernel, potato and rice flour were studied by using Rapid Visco-Analyzer (RVA). Tamarind kernel powder (TKP) varied significantly (P < 0.05) from rice and potato flours with respect to its highest protein, ash and fat contents. The results of RVA analysis indicated that pasting properties of flour/gum mixtures were dependent upon the concentration and type of the gums. Peak, breakdown and final viscosity increased with increase in gum concentration in the flour/gum mixture, but the effect was more pronounced for rice and potato flour than for TKP which showed much lower viscosity responses to all of the gums. Among the three gums studied, the increase in viscosity was significantly higher with addition of locust bean gum followed by xanthan while the lowest was observed with carboxymethylcellulose.

  3. Particle tracking microrheology of purified gastrointestinal mucins.

    PubMed

    Georgiades, Pantelis; Pudney, Paul D A; Thornton, David J; Waigh, Thomas A

    2014-04-01

    The rheological characteristics of gastric and duodenal mucin solutions, the building blocks of the mucus layer that covers the epithelia of the two organs, were investigated using particle tracking microrheology. We used biochemically well characterized purified porcine mucins (MUC5AC and MUC2) as models for human mucins, to probe their viscoelasticity as a function of mucin concentration and pH. Furthermore, we used both reducing (dithiothreitol, DTT) and chaotropic agents (guanidinium chloride and urea) to probe the mesoscopic forces that mediate the integrity of the polymer network. At neutral pH both gastric and duodenal mucins formed self-assembled semi-dilute networks above a certain critical mucin concentration (c*) with the viscosity (η) scaling as η∼c(0.53±0.08) for MUC5AC and η∼c(0.53±0.06) for MUC2, where c is the mucin concentration. Above an even higher mucin concentration threshold (ce , the entanglement concentration) reptation occurs and there is a dramatic increase in the viscosity scaling, η∼c(3.92±0.38) for MUC5AC and η∼c(5.1±0.8) for MUC2. The dynamics of the self-assembled comb polymers is examined in terms of a scaling model for flexible polyelectrolyte combs. Both duodenum and gastric mucin are found to be pH switchable gels, gelation occurring at low pHs. There is a hundred-fold increase in the elastic shear modulus once the pH is decreased. The addition of DTT, guanidinium chloride and urea disassembles both the semi-dilute and gel structures causing a large increase in the compliance (decrease in their shear moduli). Addition of the polyphenol EGCG has a reverse effect on mucin viscoelasticity, that is, it triggers a sol-gel transition in semi-dilute mucin solutions at neutral pH.

  4. Hydrogen purifier module and method for forming the same

    SciTech Connect

    DeVries, Peter David

    2012-02-07

    A hydrogen purifier utilizing a hydrogen permeable membrane, and a gas-tight seal, where the seal is uses a low temperature melting point metal, which upon heating above the melting point subsequently forms a seal alloy with adjacent metals, where the alloy has a melting point above the operational temperature of the purifier. The purifier further is constructed such that a degree of isolation exists between the metal that melts to form the seal and the active area of the purifier membrane, so that the active area of the purifier membrane is not corrupted. A method of forming a hydrogen purifier utilizing a hydrogen permeable membrane with a seal of the same type is also disclosed.

  5. Lower crosslinking density enhances functional nucleus pulposus-like matrix elaboration by human mesenchymal stem cells in carboxymethylcellulose hydrogels.

    PubMed

    Lin, Huizi A; Gupta, Michelle S; Varma, Devika M; Gilchrist, M Lane; Nicoll, Steven B

    2016-01-01

    Engineered constructs represent a promising treatment for replacement of nucleus pulposus (NP) tissue. Recently, photocrosslinked hydrogels comprised of methacrylated carboxymethylcellulose (CMC) were shown to support chondrogenic differentiation of encapsulated human mesenchymal stem cells (hMSCs) and promote accumulation of NP-like extracellular matrix (ECM). The objective of this study was to investigate the influence of CMC crosslinking density, by varying macromer concentration and modification (i.e., methacrylation) percentage, on NP-like differentiation of encapsulated hMSCs. Constructs of lower macromer concentration (2%, w/v) exhibited significantly greater collagen II accumulation, more homogeneous distribution of ECM macromolecules, and a temporal increase in mechanical properties compared to hydrogels of higher macromer concentration (4%, w/v). Constructs of higher modification percentage (25%) gave rise to significantly elevated collagen II content and the formation of cell clusters within the matrix relative to samples of lower modification percentage (10% and 15%). These differences in functional ECM accumulation and distribution are likely attributed to the distinct crosslinked network structures of the various hydrogel formulations. Overall, CMC constructs of lower macromer concentration and modification percentage were most promising as scaffolds for NP tissue engineering based on functional ECM assembly. Optimization of such hydrogel fabrication parameters may lead to the development of clinically relevant tissue-engineered NP replacements.

  6. Fabrication of Novel Hydrogel with Berberine-Enriched Carboxymethylcellulose and Hyaluronic Acid as an Anti-Inflammatory Barrier Membrane

    PubMed Central

    Huang, Yu-Chih; Huang, Kuen-Yu; Yang, Bing-Yuan

    2016-01-01

    An antiadhesion barrier membrane is an important biomaterial for protecting tissue from postsurgical complications. However, there is room to improve these membranes. Recently, carboxymethylcellulose (CMC) incorporated with hyaluronic acid (HA) as an antiadhesion barrier membrane and drug delivery system has been reported to provide excellent tissue regeneration and biocompatibility. The aim of this study was to fabricate a novel hydrogel membrane composed of berberine-enriched CMC prepared from bark of the P. amurense tree and HA (PE-CMC/HA). In vitro anti-inflammatory properties were evaluated to determine possible clinical applications. The PE-CMC/HA membranes were fabricated by mixing PE-CMC and HA as a base with the addition of polyvinyl alcohol to form a film. Tensile strength and ultramorphology of the membrane were evaluated using a universal testing machine and scanning electron microscope, respectively. Berberine content of the membrane was confirmed using a UV-Vis spectrophotometer at a wavelength of 260 nm. Anti-inflammatory property of the membrane was measured using a Griess reaction assay. Our results showed that fabricated PE-CMC/HA releases berberine at a concentration of 660 μg/ml while optimal plasticity was obtained at a 30 : 70 PE-CMC/HA ratio. The berberine-enriched PE-CMC/HA had an inhibited 60% of inflammation stimulated by LPS. These results suggest that the PE-CMC/HA membrane fabricated in this study is a useful anti-inflammatory berberine release system. PMID:28119926

  7. Pharmacokinetic and milk penetration of a difloxacin long-acting poloxamer gel formulation with carboxy-methylcellulose in lactating goats.

    PubMed

    Escudero, Elisa; Marín, Pedro; Cárceles, Carlos M; Ramírez, María J; Fernández-Varón, Emilio

    2011-04-01

    The single-dose disposition kinetics of difloxacin were determined in clinically normal lactating goats (n=6) after subcutaneous administration of a long-acting poloxamer 407 gel formulation with carboxy-methylcellulose (P407-CMC). Difloxacin concentrations were determined by high performance liquid chromatography with fluorescence detection. The concentration-time data were analysed by non-compartmental kinetic methods. Plasma and milk elimination half-lives after P407-CMC dosing were 35.19 h and 33.93 h, respectively. With this formulation, difloxacin achieved maximum plasma concentrations of 2.67±0.34 mg/L at 2.92±1.20 h and maximum milk concentrations of 2.31±0.35 mg/L at 4.00±0.00 h. The area under the curve (AUC) ratio AUC(milk)/AUC(plasma) was 0.89 after P407-CMC administration. It was concluded that a 15 mg/kg dose of difloxacin within P407-CMC would be effective against mastitis pathogens with a minimum inhibitory concentration (MIC)≤0.12 mg/L.

  8. Comparison of cytotoxicity and wound healing effect of carboxymethylcellulose and hyaluronic acid on human corneal epithelial cells

    PubMed Central

    Lee, Jong Soo; Lee, Seung Uk; Che, Cheng-Ye; Lee, Ji-Eun

    2015-01-01

    AIM To investigate the cytotoxic effect on human corneal epithelial cells (HCECs) and the ability to faciliate corneal epithelial wound healing of carboxymethylcellulose (CMC) and hyaluronic acid (HA). METHODS HCECs were exposed to 0.5% CMC (Refresh plus®, Allergan, Irvine, California, USA) and 0.1% and 0.3% HA (Kynex®, Alcon, Seoul, Korea, and Hyalein mini®, Santen, Osaka, Japan) for the period of 30min, and 4, 12, and 24h. Methyl thiazolyl tetrazolium (MTT)-based calorimetric assay was performed to assess the metabolic activity of cellular proliferation and lactate dehydrogenase (LDH) leakage assay to assess the cytotoxicity. Apoptotic response was evaluated with flow cytometric analysis and fluorescence staining with Annexin V and propiodium iodide. Cellular morphology was evaluated by inverted phase-contrast light microscopy and electron microscopy. The wound widths were measured 24h after confluent HCECs were scratch wounded. RESULTS The inhibitory effect of human corneal epithelial proliferation and cytotoxicity showed the time-dependent response but no significant effect. Apoptosis developed in flow cytometry and apoptotic cells were demonstrated in fluorescent micrograph. The damaged HCECs were detached from the bottom of the dish and showed the well-developed vacuole formations. Both CMC and HA stimulated reepithehlialization of HCECs scratched, which were more observed in CMC. CONCLUSION CMC and HA, used in artificial tear formulation, could be utilized without any significant toxic effect on HCECs. Both significantly stimulated HCEC reepithelialization of corneal wounds. PMID:25938030

  9. Salt-induced counterion condensation and related phenomena in sodium carboxymethylcellulose-sodium halide-methanol-water quaternary systems.

    PubMed

    Das, Bijan; Chatterjee, Amritendu

    2015-05-28

    Polyion-counterion interactions in sodium carboxymethylcellulose (NaCMC) in methanol-water media have been investigated conductometrically with reference to their variations with polyelectrolyte concentration, relative permittivity and the type and concentration of added electrolytes. The specific conductance data in polyelectrolyte-salt solutions were analyzed using an equation recently developed by us following the scaling description for the configuration of a polyion chain according to Dobrynin et al. Excellent quantitative agreement between the experimental results and those obtained with the new equation developed was observed. The results demonstrate that approximately 43-59% of the counterions remain free and that there has been a suppression of counterion dissociation in the presence of a salt in any given mixed solvent medium, the extent of which increases with increasing salt concentration. NaCl was found to be slightly more efficient than NaBr in suppressing the counterion-condensation in NaCMC-methanol-water systems. An increase in the amount of methanol in the media causes a reduction in the fraction of free counterions. The results further demonstrate that the monomer units experience more frictional resistance as the methanol content of the mixture increases or as the concentration of the added electrolytes increases. The results were discussed in terms of various interactions prevailing in these systems.

  10. Green synthesis, characterization and drug delivery applications of a novel silver/carboxymethylcellulose - poly(acrylamide) hydrogel nanocomposite.

    PubMed

    Gulsonbi, M; Parthasarathy, S; Bharat Raj, K; Jaisankar, V

    2016-12-01

    Biodegradable polymeric hydrogels are a unique class of macromolecular networks that can hold a large fraction of an aqueous solvent within their structures. They are particularly suitable for biomedical applications including controlled drug delivery, because of their ability to stimulate biological tissues. Many hydrogel-based networks have been designed and fabricated to meet the needs of pharmaceutical and medical fields. The investigation deals with the environment friendly synthesis of biodegradable semi interpenetrating hydrogel networks based on cross-linked poly(acrylamide) through an optimized rapid redox solution polymerization with N, N'Methylenebisacrylamide (MBA) in presence of Carboxymethylcellulose (CMC).The silver nanoparticles within hydrogel networks as nano reactors have been prepared by green synthesis via insitu reduction of silver nitrate (AgNO3) using Azadirachta Indica (Neem) plant extract under atmospheric conditions. The synthesized nanocomposites were characterised by FTIR spectroscopy, UV-Visible Spectroscopy, Scanning Electron Microscopy (SEM), Transmission electron microscopy (TEM) and Dynamic light scattering (DLS) technique. The thermal properties of the nanocomposite was analyzed by Thermogravimetric Analysis (TGA). The pH response and drug release profile of the synthesised biodegradable Silver-Hydrogel nanocomposite was investigated. Further, it was observed that physicochemical interaction between the polymeric nanocomposites and drug influences the degree of matrix swelling and therefore, its porosity and diffusion release process.

  11. Fabrication of Novel Hydrogel with Berberine-Enriched Carboxymethylcellulose and Hyaluronic Acid as an Anti-Inflammatory Barrier Membrane.

    PubMed

    Huang, Yu-Chih; Huang, Kuen-Yu; Yang, Bing-Yuan; Ko, Chun-Han; Huang, Haw-Ming

    2016-01-01

    An antiadhesion barrier membrane is an important biomaterial for protecting tissue from postsurgical complications. However, there is room to improve these membranes. Recently, carboxymethylcellulose (CMC) incorporated with hyaluronic acid (HA) as an antiadhesion barrier membrane and drug delivery system has been reported to provide excellent tissue regeneration and biocompatibility. The aim of this study was to fabricate a novel hydrogel membrane composed of berberine-enriched CMC prepared from bark of the P. amurense tree and HA (PE-CMC/HA). In vitro anti-inflammatory properties were evaluated to determine possible clinical applications. The PE-CMC/HA membranes were fabricated by mixing PE-CMC and HA as a base with the addition of polyvinyl alcohol to form a film. Tensile strength and ultramorphology of the membrane were evaluated using a universal testing machine and scanning electron microscope, respectively. Berberine content of the membrane was confirmed using a UV-Vis spectrophotometer at a wavelength of 260 nm. Anti-inflammatory property of the membrane was measured using a Griess reaction assay. Our results showed that fabricated PE-CMC/HA releases berberine at a concentration of 660 μg/ml while optimal plasticity was obtained at a 30 : 70 PE-CMC/HA ratio. The berberine-enriched PE-CMC/HA had an inhibited 60% of inflammation stimulated by LPS. These results suggest that the PE-CMC/HA membrane fabricated in this study is a useful anti-inflammatory berberine release system.

  12. Toxoids of Pseudomonas aeruginosa exotoxin-A: photoaffinity inactivation of purified toxin and purified toxin derivatives.

    PubMed Central

    Callahan, L T; Martinez, D; Marburg, S; Tolman, R L; Galloway, D R

    1984-01-01

    For the preparation of greatly detoxified but highly immunogenic toxoids, two enzymatically active, low-toxicity derivatives of Pseudomonas aeruginosa exotoxin-A were further inactivated by photoaffinity labeling. These derivatives were formed during toxin purification, when a relatively crude toxin preparation was concentrated by ammonium sulfate precipitation and subsequently dialyzed. These derivatives, designated peak-1 protein (PK-1) and peak-2 protein (PK-2) were antigenically indistinguishable from native toxin, but had isoelectric points (5.00 and 4.90, respectively) that were different from that of the native toxin (4.95). Although the enzymatic activities and molecular weights of PK-1 and PK-2 were similar to those of native toxin, their toxicities were greatly reduced (ca. 500-fold). Photoaffinity labeling of fully active toxin-A, purified by a process which limits the formation of these derivatives, decreased its enzymatic activity (ca. 30-fold) and toxicity (ca. 100-fold). Likewise, photoaffinity labeling of purified PK-1 and PK-2 decreased their enzymatic activities and toxicities (ca. 30-fold and 100-fold, respectively) and, thus, yielded toxoids that were ca. 50,000-fold less toxic than unpurified native toxin. These toxoids were irreversibly detoxified and highly immunogenic during 9 months of storage at 4 degrees C. Images PMID:6321348

  13. 21 CFR 880.6500 - Medical ultraviolet air purifier.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Medical ultraviolet air purifier. 880.6500 Section... (CONTINUED) MEDICAL DEVICES GENERAL HOSPITAL AND PERSONAL USE DEVICES General Hospital and Personal Use Miscellaneous Devices § 880.6500 Medical ultraviolet air purifier. (a) Identification. A medical ultraviolet...

  14. Portable self-contained solar powered water purifier

    SciTech Connect

    Sherman, M.

    1991-10-22

    This patent describes a portable self-contained solar powered water purifier. It comprises housing means for buoyantly supporting the purifier; solar cell means supported by the housing means above water to be treated; purification means depending from the housing means so as to be positioned in water to be treated and including sacrificial anode means providing ionized metallic ions for purifying the water and cathode means providing abstraction of electrons to facilitate the release of oxygen into the water; means for electrically connecting the solar cell means to the electrolytic purification means to enable the electrolytic purification means to purify water when the purifier is placed therein; and diode means for preventing reverse current flow between the anode means and cathode means.

  15. Aspects on the interaction between sodium carboxymethylcellulose and calcium carbonate and the relationship to specific site adsorption.

    PubMed

    Backfolk, Kaj; Lagerge, Serge; Rosenholm, Jarl B; Eklund, Dan

    2002-04-01

    The mechanisms of adsorption and association for sodium carboxymethylcellulose (NaCMC) in calcium carbonate suspensions have been determined from isothermal calorimetry and adsorption measurements. The equilibrium adsorption isotherms were determined by two different methods of separation; a depletion method and a serum exchange method. The enthalpy of dilution for NaCMC was determined on supernatants obtained from the calcium carbonate suspensions in order to investigate the interaction between NaCMC and dissolved species from the mineral. For comparison, NaCMC was injected into CaCl(2) solutions in order to determine the role of calcium ions in the adsorption process. The initial part of the adsorption isotherm showed a quasi-infinite slope indicating a high affinity for the NaCMC to the calcium carbonate surface, which was significantly reduced when anionic sodium polyacrylate was preadsorbed onto the calcium carbonate implying competitive adsorption. An endothermic enthalpy change was observed between the NaCMC and the calcium carbonate surface, suggesting attachment of the carboxylic acid groups onto the hydrated calcium sites. A similar endothermic enthalpy was observed when NaCMC was injected into CaCl(2) solutions or supernatants obtained from the calcium carbonate suspensions, indicating a complexation of carboxylic acid groups and hydrated calcium ions. It was concluded that the mechanisms of interaction of NaCMC in calcium carbonate suspensions are primarily an association between NaCMC and Lewis acid sites on the calcium carbonate surface and the formation of NaCMC-Ca(2+) complexes in the bulk solution, both of which will be affected by the amount of anionic sodium polyacrylate present.

  16. Antisera against electrophoretically purified tubulin stimulate colchicine-binding activity.

    PubMed Central

    Aubin, J E; Subrahmanyan, L; Kalnins, V I; Ling, V

    1976-01-01

    Several rabbit antisera have been prepared against reduced and alkylated, electrophoretically purified tubulin isolated from chick brain. These antisera give a single precipitin line in Ouchterlony double diffusion plates when tested against partially purified tubulin, and label specifically microtubule- and tubulin-containing structures, such as mitotic spindles, cilia, and vinblastine-induced crystals, in a variety of cells. The same antisera also display the unique ability to stimulate the colchicine-binding activity of tubulin preparations from chick brain and Chinese hamster ovary tissue culture cells. This specific stimulation of colchicine binding activity is also obtained with the gamma globulin fractions purified by ammonium sulfate precipitation of these antisera. Images PMID:57619

  17. Vacuolar H(+)-pyrophosphatase purified from pear fruit.

    PubMed

    Suzuki, Y; Kanayama, Y; Shiratake, K; Yamaki, S

    1999-02-01

    A vacuolar H(+)-translocating inorganic pyrophosphatase was purified from pear fruit through selective detergent treatments, Superose 6 and Mono Q column chromatography. The specific activity of the purified enzyme was 850 mumol h-1 mg protein-1. The Mr of V-PPase was 66 kDa by SDS-PAGE and the polypeptide cross-reacted with the antiserum against V-PPase of mung bean. The purified V-PPase was stimulated by potassium and inhibited by calcium and N, N'-dicyclohexylcarbodiimide.

  18. Study of gas purifiers for the CMS RPC detector

    NASA Astrophysics Data System (ADS)

    Benussi, L.; Bianco, S.; Colafranceschi, S.; Fabbri, F. L.; Felli, F.; Ferrini, M.; Giardoni, M.; Greci, T.; Paolozzi, A.; Passamonti, L.; Piccolo, D.; Pierluigi, D.; Russo, A.; Saviano, G.; Buontempo, S.; Cimmino, A.; de Gruttola, M.; Fabozzi, F.; Iorio, A. O. M.; Lista, L.; Paolucci, P.; Baesso, P.; Belli, G.; Pagano, D.; Ratti, S. P.; Vicini, A.; Vitulo, P.; Viviani, C.; Guida, R.; Sharma, A.

    2012-01-01

    The CMS RPC muon detector utilizes a gas recirculation system called closed loop (CL) to cope with large gas mixture volumes and costs. A systematic study of CL gas purifiers has been carried out over 400 days between July 2008 and August 2009 at CERN in a low-radiation test area, with the use of RPC chambers with currents monitoring, and gas analysis sampling points. The study aimed to fully clarify the presence of pollutants, the chemistry of purifiers used in the CL, and the regeneration procedure. Preliminary results on contaminants release and purifier characterization are reported.

  19. A recombinant envelope protein from Dengue virus purified by IMAC is bioequivalent with its immune-affinity chromatography purified counterpart.

    PubMed

    Hermida, L; Rodríguez, R; Lazo, L; López, C; Márquez, G; Páez, R; Suárez, C; Espinosa, R; García, J; Guzmán, G; Guillén, G

    2002-03-28

    Semi-purified DEN-4 envelope protein, obtained in Pichia pastoris, was capable of generating neutralising and protecting antibodies after immunisation in mice. Here we compared two purification processes of this recombinant protein using two chromatographic steps: immune-affinity chromatography and immobilised metal ion adsorption chromatography (IMAC). The protein purified by both methods produced functional antibodies reflected by titres of haemagglutination inhibition and neutralisation. IMAC could be used as an alternative for high scale purification.

  20. Duration of TGF-β3 Exposure Impacts the Chondrogenic Maturation of Human MSCs in Photocrosslinked Carboxymethylcellulose Hydrogels.

    PubMed

    Gupta, Michelle S; Nicoll, Steven B

    2015-05-01

    Intervertebral disc (IVD) herniation can be caused by both degeneration and traumatic injury, ultimately resulting in back pain or sciatica due to disc protrusion. Replacement of the nucleus pulposus (NP) tissue during surgical intervention post herniation could improve the long-term stability of the functional spinal unit. Tissue engineering strategies may potentially restore both biological and mechanical function of the NP. Recently, photocrosslinked carboxymethylcellulose (CMC) hydrogels were shown to support chondrogenic, NP-like extracellular matrix (ECM) elaboration by human mesenchymal stromal cells (hMSCs) when supplemented with TGF-β3. However, long-term preconditioning with soluble growth factors in vitro or the use of sustained growth factor delivery vehicles in vivo can be expensive and difficult to control. Transient supplementation with growth factors has been shown to maintain or improve maturation of tissue-engineered constructs. The objective of this study was to evaluate the influence of TGF-β3 exposure time on hydrogel bulk properties and NP-like matrix elaboration in hMSC-laden CMC hydrogels. Constructs were exposed to TGF-β3 for 2 weeks (Transient), 8 weeks (Continuous) or 0 weeks (controls). After 8 weeks of culture, both the Transient and Continuous groups exhibited increased ECM accumulation compared to 2 weeks and controls. The Transient group displayed significantly greater accumulation of collagens I and II, while GAG content was significantly higher in the Continuous group by 8 weeks. Distribution of ECM was more homogeneous in the Continuous group, while the Transient group exhibited more concentrated accumulation in the periphery of the hydrogel by 8 weeks. Mechanical properties improved over time in both groups, however, Continuous constructs demonstrated significantly more robust mechanical properties (equilibrium modulus and peak stress) compared to Transient gels at 8 weeks. Although the functional properties of

  1. Effectiveness and short-term safety of modified sodium hyaluronic acid-carboxymethylcellulose at cesarean delivery: A randomized trial

    PubMed Central

    KIEFER, Daniel G.; MUSCAT, Jolene C.; SANTORELLI, Jarrett; CHAVEZ, Martin R.; ANANTH, Cande V.; SMULIAN, John C.; VINTZILEOS, Anthony M.

    2016-01-01

    Background The rising cesarean birth rate has drawn attention to risks associated with repeat cesarean birth. Prevention of adhesions with adhesion barriers has been promoted as a way to decrease operative difficulty. However, robust data demonstrating effectiveness of such interventions are lacking. Objective We report data from a multicenter trial designed to evaluate the short-term safety and effectiveness of a modified sodium hyaluronate-carboxymethylcellulose absorbable (HACMC) for reduction of adhesions following cesarean delivery. Methods Patients that underwent primary or repeat cesarean delivery were included in this multicenter, single-blinded (patient), randomized controlled trial. Patients were randomized into either HA-CMC (N=380) or no-treatment group (N=373). No other modifications to their treatment were part of the protocol. Short-term safety data was collected following randomization. The location and density of adhesions (primary outcome) were assessed at their subsequent delivery using a validated tool, which can also be used to derive an adhesion score, that ranges from 0–12. Results No differences in baseline characteristics, post-operative course, or incidence of complications between the groups following randomization were noted. Eighty patients from the HA-CMC group and 92 controls returned for subsequent deliveries. Adhesions in any location were reported in 75.6% of the HA-CMC group and 75.9% of the controls (P=0.99). There was no significant difference in the median adhesion score; 2 (range 0–10) for the HACMC group vs. 2 (range 0–8) for the control group (P=0.65). One third of the HA-CMC patients met the definition for severe adhesions (adhesion score >4) compared to 15.5% in the control group (P=0.052). There were no significant differences in the time from incision to delivery (P=0.56). Uterine dehiscence in the next pregnancy was reported in 2 patients in HA-CMC group versus 1 in the control (P=0.60). Conclusions Although we

  2. Strong purifying selection at genes escaping X chromosome inactivation.

    PubMed

    Park, Chungoo; Carrel, Laura; Makova, Kateryna D

    2010-11-01

    To achieve dosage balance of X-linked genes between mammalian males and females, one female X chromosome becomes inactivated. However, approximately 15% of genes on this inactivated chromosome escape X chromosome inactivation (XCI). Here, using a chromosome-wide analysis of primate X-linked orthologs, we test a hypothesis that such genes evolve under a unique selective pressure. We find that escape genes are subject to stronger purifying selection than inactivated genes and that positive selection does not significantly affect the evolution of these genes. The strength of selection does not differ between escape genes with similar versus different expression levels in males versus females. Intriguingly, escape genes possessing Y homologs evolve under the strongest purifying selection. We also found evidence of stronger conservation in gene expression levels in escape than inactivated genes. We hypothesize that divergence in function and expression between X and Y gametologs is driving such strong purifying selection for escape genes.

  3. Functional analysis and regulation of purified connexin hemichannels

    PubMed Central

    Fiori, Mariana C.; Reuss, Luis; Cuello, Luis G.; Altenberg, Guillermo A.

    2014-01-01

    Gap-junction channels (GJCs) are aqueous channels that communicate adjacent cells. They are formed by head-to-head association of two hemichannels (HCs), one from each of the adjacent cells. Functional HCs are connexin hexamers composed of one or more connexin isoforms. Deafness is the most frequent sensineural disorder, and mutations of Cx26 are the most common cause of genetic deafness. Cx43 is the most ubiquitous connexin, expressed in many organs, tissues, and cell types, including heart, brain, and kidney. Alterations in its expression and function play important roles in the pathophysiology of very frequent medical problems such as those related to cardiac and brain ischemia. There is extensive information on the relationship between phosphorylation and Cx43 targeting, location, and function from experiments in cells and organs in normal and pathological conditions. However, the molecular mechanisms of Cx43 regulation by phosphorylation are hard to tackle in complex systems. Here, we present the use of purified HCs as a model for functional and structural studies. Cx26 and Cx43 are the only isoforms that have been purified, reconstituted, and subjected to functional and structural analysis. Purified Cx26 and Cx43 HCs have properties compatible with those demonstrated in cells, and present methodologies for the functional analysis of purified HCs reconstituted in liposomes. We show that phosphorylation of serine 368 by PKC produces a partial closure of the Cx43 HCs, changing solute selectivity. We also present evidence that the effect of phosphorylation is highly cooperative, requiring modification of several connexin subunits, and that phosphorylation of serine 368 elicits conformational changes in the purified HCs. The use of purified HCs is starting to provide critical data to understand the regulation of HCs at the molecular level. PMID:24611052

  4. 21 CFR 880.6710 - Medical ultraviolet water purifier.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Medical ultraviolet water purifier. 880.6710 Section 880.6710 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GENERAL HOSPITAL AND PERSONAL USE DEVICES General Hospital and Personal Use Miscellaneous Devices § 880.6710...

  5. Obtaining partial purified xylose reductase from Candida guilliermondii

    PubMed Central

    Tomotani, Ester Junko; de Arruda, Priscila Vaz; Vitolo, Michele; de Almeida Felipe, Maria das Graças

    2009-01-01

    The enzymatic bioconversion of xylose into xylitol by xylose reductase (XR) is an alternative for chemical and microbiological processes. The partial purified XR was obtained by using the following three procedures: an agarose column, a membrane reactor or an Amicon Ultra-15 50K Centrifugal Filter device at yields of 40%, 7% and 67%, respectively. PMID:24031408

  6. Small-Scale Evaluation of the Expeditionary Unit Water Purifier

    EPA Science Inventory

    The U.S. EPA’s Technology Testing and Evaluation Program has been charged by EPA to evaluate the performance of commercially available water security-related technologies. Throughout 2007, an evaluation the Expeditionary Unit Water Purifier (EUWP), a mobile water treatment techno...

  7. Strong Purifying Selection in Transmission of Mammalian Mitochondrial DNA

    PubMed Central

    Stewart, James Bruce; Freyer, Christoph; Elson, Joanna L; Wredenberg, Anna; Cansu, Zekiye; Trifunovic, Aleksandra; Larsson, Nils-Göran

    2008-01-01

    There is an intense debate concerning whether selection or demographics has been most important in shaping the sequence variation observed in modern human mitochondrial DNA (mtDNA). Purifying selection is thought to be important in shaping mtDNA sequence evolution, but the strength of this selection has been debated, mainly due to the threshold effect of pathogenic mtDNA mutations and an observed excess of new mtDNA mutations in human population data. We experimentally addressed this issue by studying the maternal transmission of random mtDNA mutations in mtDNA mutator mice expressing a proofreading-deficient mitochondrial DNA polymerase. We report a rapid and strong elimination of nonsynonymous changes in protein-coding genes; the hallmark of purifying selection. There are striking similarities between the mutational patterns in our experimental mouse system and human mtDNA polymorphisms. These data show strong purifying selection against mutations within mtDNA protein-coding genes. To our knowledge, our study presents the first direct experimental observations of the fate of random mtDNA mutations in the mammalian germ line and demonstrates the importance of purifying selection in shaping mitochondrial sequence diversity. PMID:18232733

  8. Effect of human and simulated gastric juices on the digestion of whey proteins and carboxymethylcellulose-stabilised O/W emulsions.

    PubMed

    Malinauskytė, Ernesta; Ramanauskaitė, Jovita; Leskauskaitė, Daiva; Devold, Tove G; Schüller, Reidar B; Vegarud, Gerd E

    2014-12-15

    In this study, we analysed the impact of carboxymethylcellulose (CMC) on lipid digestion and physicochemical properties of whey proteins (WP)-stabilised emulsions during in vitro digestion with either artificial or human gastrointestinal juices. The emulsions were made by adsorbing WP on the fat droplets and subsequently adding CMC, which does not interact with the adsorbed proteins. The limited hydrolysis of lipids and their higher physical stability was recorded for WP-stabilised emulsions in the presence of CMC under simulated gastrointestinal conditions. The possible mechanism by which CMC lowers the digestion of WP-stabilised emulsions is related to the limited interaction of fat droplets with gastrointestinal fluids due to the extended thickening network formed by CMC in the continuous phase. The digestion of WP- and CMC-stabilised emulsions in the in vitro model with human gastric fluids led to greater lipid hydrolysis, although the enzymatic activity in both in vitro models was observed at the same level.

  9. The Use of a Hyaluronic Acid–Carboxymethylcellulose Membrane as an Adhesion Barrier during Nipple Delay before Nipple-preserving Mastectomy

    PubMed Central

    Jablonka, Eric M.; Harmaty, Marco A.; Torina, Philip J.

    2016-01-01

    Summary: The desire for nipple-preserving mastectomies has increased over the past decade as studies have proven that the procedure has comparable oncologic risk as a traditional mastectomy in both therapeutic and prophylactic cases. Partial or complete nipple necrosis is a well-known complication of this procedure with rates in the literature ranging between 1% and 9%. In high-risk patients, surgeons are performing a delay procedure before the mastectomy to help improve nipple vascularity and decrease necrosis rates. We present a technique of using a short-lasting bioresorbable hyaluronate–carboxymethylcellulose membrane (Seprafilm; Genzyme, Cambridge, Mass.) during the delay procedure as an interpositional sheet, which prevents adhesion of the anterior breast skin from the underlying gland to both block revascularization to improve nipple perfusion and prevent the need for redissection of the previously created plane during the final mastectomy. PMID:28293518

  10. Properties of purified recombinant human polyamine oxidase, PAOh1/SMO.

    PubMed

    Wang, Yanlin; Murray-Stewart, Tracy; Devereux, Wendy; Hacker, Amy; Frydman, Benjamin; Woster, Patrick M; Casero, Robert A

    2003-05-16

    The discovery of an inducible oxidase whose apparent substrate preference is spermine indicates that polyamine catabolism is more complex than that originally proposed. To facilitate the study of this enzyme, the purification and characterization of the recombinant human PAOh1/SMO polyamine oxidase are reported. Purified PAOh1/SMO oxidizes both spermine (K(m)=1.6 microM) and N(1)-acetylspermine (K(m)=51 microM), but does not oxidize spermidine. The purified human enzyme also does not oxidize eight representative antitumor polyamine analogues; however, specific oligamine analogues were found to be potent inhibitors of the oxidation of spermine by PAOh1/SMO. The results of these studies are consistent with the hypothesis that PAOh1/SMO represents a new addition to the polyamine metabolic pathway that may represent a new target for antineoplastic drug development.

  11. Hydrolysis of cellulose by purified cellulase components: Synergistic effects

    SciTech Connect

    Woodward, J.; Lee, N.E.

    1987-01-01

    The hydrolysis of cellulose by purified cellulase components is reported. The adsorption of purified cellobiohydrolases (CBH I and II) and endoglucanases (EG I and II) from Trichoderma reesei strain L27 to microcrystalline cellulose (Avicel) has been studied. Scatchard analysis of the adsorption data gave the maximum amount of each component that bound to Avicel at saturation. Hydrolysis of Avicel was thus carried out by saturating and non-saturating (50% saturation) concentrations of cellulase components alone and in combination with each other, and it was found that the greatest amount of synergism between them was observed when Avicel was incubated with non-saturating concentrations of enzyme. Synergism was observed between CBH I and CBH II, as well as between EG I and CBH I; however, inhibition of hydrolysis occurred using a combination of EG I and EG II. Synergism between cellulase components may be significant during cellulose hydrolysis only when non-saturating enzyme concentrations are used. 4 refs., 3 figs.

  12. Purifying food-grade, naturally occurring CO/sub 2/

    SciTech Connect

    Nobles, J.E.; Stancik, J.W.

    1983-12-26

    Technology to purify naturally occurring CO/sub 2/ into products suitable for enhanced oil recovery (EOR) and food grade CO/sub 2/ markets has been commercially demonstrated in a 20 MMscfd (1,100 ton/day) CO/sub 2/ processing facility owned and operated by Columbia Hydrocarbon Corp. The unit demonstrates that it is technically and economically feasible to process low grade natural gas to produce pipeline specification natural gas, raw CO/sub 2/, and food grade CO/sub 2/. The Selexol treating facility is located on a reclaimed coal strip mine with very restricted plot area. Only compression and dehydration equipment for the CO/sub 2/ is located there. The CO/sub 2/ is transported by pipeline to the Marmet site, approx. 7 miles away. This site contains the facilities for purifying the CO/sub 2/ and for storage.

  13. Application of paramagnetic beads for purifying Bacillus anthracis protective antigen.

    PubMed

    Zarzecka, A; Bartoszcze, M

    2006-10-01

    Paramagnetic beads coated with Protein G and Tosylactivated-280 dynabeads have been used to purify Bacillus anthracis protective antigen from a liquid culture. The obtained protein was used in the enzyme-linked immunosorbent assay test to detect B. anthracis protective antigen antibodies in human sera collected from immunized individuals. The purification method using paramagnetic beads is very effective. It is fast, easy and may be carried out practically in any laboratory.

  14. Thermostable purified endoglucanase from thermophilic bacterium acidothermus cellulolyticus

    DOEpatents

    Tucker, Melvin P.; Grohmann, Karel; Himmel, Michael E.; Mohagheghi, Ali

    1992-01-01

    A substantially purified high molecular weight cellulase enzyme having a molecular weight of between about 156,000 to about 203,400 daltons isolated from the bacterium Acidothermus cellulolyticus (ATCC 43068) and a method of producing it are disclosed. The enzyme is water soluble, possesses both C.sub.1 and C.sub.x types of enzymatic activity, has a high degree of stability toward heat and exhibits both a high optimum temperature activity and high inactivation characteristics.

  15. Thermostable purified endoglucanase II from Acidothermus cellulolyticus ATCC

    DOEpatents

    Adney, William S.; Thomas, Steven R.; Nieves, Rafael A.; Himmel, Michael E.

    1994-01-01

    A purified low molecular weight endoglucanase II from Acidothermus cellulolyticus (ATCC 43068) is disclosed. The endoglucanase is water soluble, possesses both C.sub.1, and C.sub.x types of enzyme activity, a high degree of stability toward heat, and exhibits optimum temperature activity at about 81.degree. C. at pH's from about 2 to about 9, and at a inactivation temperature of about 100.degree. C. at pH's from about 2 to about 9.

  16. Thermostable purified endoglucanase II from Acidothermus cellulolyticus ATCC

    DOEpatents

    Adney, W.S.; Thomas, S.R.; Nieves, R.A.; Himmel, M.E.

    1994-11-22

    A purified low molecular weight endoglucanase II from Acidothermus cellulolyticus (ATCC 43068) is disclosed. The endoglucanase is water soluble, possesses both C[sub 1], and C[sub x] types of enzyme activity, a high degree of stability toward heat, and exhibits optimum temperature activity at about 81 C at pH's from about 2 to about 9, and at a inactivation temperature of about 100 C at pH's from about 2 to about 9. 9 figs.

  17. [Proteomic Analyses of Purified Particles of the Rabies Virus].

    PubMed

    Tu, Zhongzhong; Gong, Wenjie; Zhang, Yan; Feng, Ye; Li, Nan; Tu, Changchun

    2015-05-01

    The rabies virus (RABV) is an enveloped RNA virus. It mainly damages the central nervous system and causes anencephaly in mammals and humans. There is now compelling evidence that enveloped virions released from infected cells can carry many host proteins, some of which may play an important part in viral replication. Several host proteins have been reported to be incorporated into RABV particles. However, a systematic study to reveal the proteomics of RABV particles has not been conducted. In the present study, after virus culture and purification by sucrose density gradient ultracentrifugation, a proteomics approach was used to analyze the protein composition of purified RABV particles to understand the molecular mechanisms of virus-cell interactions. Fifty host proteins, along with five virus-encoded structural proteins, were identified in purified RABV particles. These proteins could be classified into ten categories according to function: intracellular trafficking (14%), molecular chaperone (12%), cytoskeletal (24%), signal transduction (8%), transcription regulation (12%), calcium ion-binding (6%), enzyme binding (6%), metabolic process (2%), ubiquitin (2%) and other (14%). Of these, four proteins (beta-actin, p-tubulin, Cofilin, Hsc70) were validated by western blotting to be present in purified RABV particles. This novel study of the composition of host proteins in RABV particles may aid investigation of the mechanism of RABV replication.

  18. THE PREPARATION AND PROPERTIES OF HIGHLY PURIFIED ASCORBIC ACID OXIDASE

    PubMed Central

    Powers, Wendell H.; Lewis, Stanley; Dawson, Charles R.

    1944-01-01

    1. A method is described for the preparation of a highly purified ascorbic acid oxidase containing 0.24 per cent copper. 2. Using comparable activity measurements, this oxidase is about one and a half times as active on a dry weight basis as the hitherto most highly purified preparation described by Lovett-Janison and Nelson. The latter contained 0.15 per cent copper. 3. The oxidase activity is proportional to the copper content and the proportionality factor is the same as that reported by Lovett-Janison and Nelson. 4. When dialyzed free of salt, the blue concentrated oxidase solutions precipitate a dark green-blue protein which carries the activity. This may be prevented by keeping the concentrated solutions about 0.1 M in Na2HPO4. 5. When highly diluted for activity measurements the oxidase rapidly loses activity (irreversibly) previous to the measurement, unless the dilution is made with a dilute inert protein (gelatin) solution. Therefore activity values obtained using such gelatin-stabilized dilute solutions of the oxidase run considerably higher than values obtained by the Lovett-Janison and Nelson technique. 6. The effect of pH and substrate concentration on the activity of the purified oxidase in the presence and absence of inert protein was studied. PMID:19873382

  19. Generation of antisera to purified prions in lipid rafts.

    PubMed

    Hnasko, Robert; Serban, Ana V; Carlson, George; Prusiner, Stanley B; Stanker, Larry H

    2010-01-01

    Prion diseases are fatal neurodegenerative disorders caused by prion proteins (PrP). Infectious prions accumulate in the brain through a template-mediated conformational conversion of endogenous PrP(C) into alternately folded PrP(Sc). Immunoassays toward pre-clinical detection of infectious PrP(Sc) have been confounded by low-level prion accumulation in non-neuronal tissue and the lack of PrP(Sc) selective antibodies. We report a method to purify infectious PrP(Sc) from biological tissues for use as an immunogen and sample enrichment for increased immunoassay sensitivity. Significant prion enrichment is accomplished by sucrose gradient centrifugation of infected tissue and isolation with detergent resistant membranes from lipid rafts (DRMs). At equivalent protein concentration a 50-fold increase in detectable PrP(Sc) was observed in DRM fractions relative to crude brain by direct ELISA. Sequential purification steps result in increased specific infectivity (DRM <20-fold and purified DRM immunogen <40-fold) relative to 1% crude brain homogenate. Purification of PrP(Sc) from DRM was accomplished using phosphotungstic acid protein precipitation after proteinase-K (PK) digestion followed by size exclusion chromatography to separate PK and residual protein fragments from larger prion aggregates. Immunization with purified PrP(Sc) antigen was performed using wild-type (wt) and Prnp(0/0) mice, both on Balb/cJ background. A robust immune response against PrP(Sc) was observed in all inoculated Prnp(0/0) mice resulting in antisera containing high-titer antibodies against prion protein. Antisera from these mice recognized both PrP(C) and PrP(Sc), while binding to other brain-derived protein was not observed. In contrast, the PrP(Sc) inoculum was non-immunogenic in wt mice and antisera showed no reactivity with PrP or any other protein.

  20. Generation of antisera to purified prions in lipid rafts

    PubMed Central

    Hnasko, Robert; Serban, Ana V; Carlson, George; Prusiner, Stanley B

    2010-01-01

    Prion diseases are fatal neurodegenerative disorders caused by prion proteins (PrP). Infectious prions accumulate in the brain through a template-mediated conformational conversion of endogenous PrPC into alternately folded PrPSc. Immunoassays toward pre-clinical detection of infectious PrPSc have been confounded by low-level prion accumulation in non-neuronal tissue and the lack of PrPSc selective antibodies. We report a method to purify infectious PrPSc from biological tissues for use as an immunogen and sample enrichment for increased immunoassay sensitivity. Significant prion enrichment is accomplished by sucrose gradient centrifugation of infected tissue and isolation with detergent resistant membranes from lipid rafts (DRMs). At equivalent protein concentration a 50-fold increase in detectable PrPSc was observed in DRM fractions relative to crude brain by direct ELISA. Sequential purification steps result in increased specific infectivity (DRM >20-fold and purified DRM immunogen >40-fold) relative to 1% crude brain homogenate. Purification of PrPSc from DRM was accomplished using phosphotungstic acid protein precipitation after proteinase-K (PK) digestion followed by size exclusion chromatography to separate PK and residual protein fragments from larger prion aggregates. Immunization with purified PrPSc antigen was performed using wild-type (wt) and Prnp0/0 mice, both on Balb/cJ background. A robust immune response against PrPSc was observed in all inoculated Prnp0/0 mice resulting in antisera containing high-titer antibodies against prion protein. Antisera from these mice recognized both PrPC and PrPSc, while binding to other brain-derived protein was not observed. In contrast, the PrPSc inoculum was non-immunogenic in wt mice and antisera showed no reactivity with PrP or any other protein. PMID:20647769

  1. Comparative study of crude and purified cellulose from wheat straw.

    PubMed

    Sun, Xiao-Feng; Sun, Run-Cang; Su, Yinquan; Sun, Jing-Xia

    2004-02-25

    A sequential totally chlorine-free procedure for isolation of cellulose from wheat straw was proposed in this study. The dewaxed straw was pretreated with 0.5 M NaOH in 60% methanol at 60 degrees C for 2.5 h under ultrasonic irradiation for 0-35 min and sequentially posttreated with 2% H(2)O(2)-0.2% TAED at pH 11.8 for 12 h at 48 degrees C, which together solubilized 85.3-86.1% of the original hemicelluloses and 91.7-93.2% of the original lignin, respectively. The yield of crude cellulose ranged between 46.2 and 49.2% on a dry weight basis related to wheat straw, which contained 11.2-12.2% residual hemicelluloses and 2.5-2.9% remaining lignin. Further treatment of the corresponding crude cellulosic preparations with 80% acetic acid-70% nitric acid under the condition given yielded 36.8-37.7% of the purified cellulose, which contained minor amounts of bound hemicelluloses (2.5-2.8%) and was relatively free of associated lignin (0.1-0.2%). The isolated crude and purified cellulose samples were comparatively studied by FT-IR and CP/MAS (13)C NMR spectroscopy, and the relative crystallinity was also estimated. The final stage treatment with 80% acetic acid-70% nitric acid decreased the hemicelluloses and lignin associated in the crude cellulose but led to 3.1-5.4% degradation of the original cellulose; in addition, the purity of the obtained cellulose was high. However, it was found that the final stage treatment is not severe enough to cause decrystallization of cellulose. The thermal stability of the purified cellulose is higher than that of the corresponding crude cellulose.

  2. Purifying protein complexes for mass spectrometry: applications to protein translation.

    PubMed

    Link, Andrew J; Fleischer, Tracey C; Weaver, Connie M; Gerbasi, Vincent R; Jennings, Jennifer L

    2005-03-01

    Proteins control and mediate most of the biological activities in the cell. In most cases, proteins either interact with regulatory proteins or function in large molecular assemblies to carryout biological processes. Understanding the functions of individual proteins requires the identification of these interacting proteins. With its speed and sensitivity, mass spectrometry has become the dominant method for identifying components of protein complexes. This article reviews and discusses various approaches to purify protein complexes and analyze the proteins using mass spectrometry. As examples, methods to isolate and analyze protein complexes responsible for the translation of messenger RNAs into polypeptides are described.

  3. [Importance of air purifiers in the prevention of respiratory allergy].

    PubMed

    Sabbah, A

    2000-06-01

    Prevention of respiratory allergy and thus diminution of its prevalence consists of the application not only of preventative measures on the allergens themselves, but also on atmospheric pollutants such as NO, CO and diesel particles. These modify the allergens and increase synthesis of allergen-specific IgE. Use of air purifiers, on condition that they have HEPA or ULPA filters may contribute to reduction of allergenicity linked to pollutants and are one of the not-negligible means of protection from respiratory allergy and asthma in particular.

  4. Thermostable purified endoglucanas from acidothermus cellulolyticus ATCC 43068

    DOEpatents

    Himmel, Michael E.; Adney, William S.; Tucker, Melvin P.; Grohmann, Karel

    1994-01-01

    A purified low molecular weight cellulase endoglucanase I having a molecular weight of between about 57,420 to about 74,580 daltons from Acidothermus cellulolyticus (ATCC 43068). The cellulase is water soluble, possesses both C.sub.1 and C.sub.x types of enzyme activity, a high degree of stability toward heat, and exhibits optimum temperature activity at about 83.degree. C. at pH's from about 2 to about 9, and in inactivation temperature of about 110.degree. C. at pH's from about 2 to about 9.

  5. Thermostable purified endoglucanase from Acidothermus cellulolyticus ATCC 43068

    DOEpatents

    Himmel, M.E.; Adney, W.S.; Tucker, M.P.; Grohmann, K.

    1994-01-04

    A purified low molecular weight cellulase endoglucanase I having a molecular weight of between about 57,420 to about 74,580 daltons from Acidothermus cellulolyticus (ATCC 43068) is presented. The cellulase is water soluble, possesses both C[sub 1] and C[sub x] types of enzyme activity, a high degree of stability toward heat, and exhibits optimum temperature activity at about 83 C at pH's from about 2 to about 9, and in inactivation temperature of about 110 C at pH's from about 2 to about 9. 7 figures.

  6. Growth of purified astrocytes in a chemically defined medium

    SciTech Connect

    Morrison, R.S.; De Vellis, J.

    1981-11-01

    Astrocytes purified from primary cultures of neonatal rat cerebrum can not be grown in a synthetic medium supplemented with putrescine, prostaglandin F/sub 2//sub ..cap alpha../, insulin, fibroblast growth factor, and hydrocortisone. These five supplements have a marked synergistic effect on growth when used in combination but have little effect when used individually. Astrocytes grown in the defined medium exhibit dramatic changes in morphological characteristics in comparison to cells grown in serum-free or serum-supplemented medium. In addition, these cells express the astrocyte-specific marker glial fibrillary acidic protein and are estimated by several criteria to be greater than 95% astrocytes.

  7. Spectrophotometric assay for amikacin using purified kanamycin acetyltransferase.

    PubMed

    Scarbrough, E; Williams, J W; Northrop, D B

    1979-08-01

    A rapid spectrophotometric assay has been developed for measuring the concentrations of amikacin and related antibiotics in serum. The assay uses a purified enzyme from R-factor E. coli which acetylates amikacin with the production of coenzyme A, the latter in turn being reacted with a sulfhydryl reagent to produce stoichiometric amounts of a sensitive chromophore, that is measured in the visible spectrum. The system complements an earlier assay for gentamicin-related antibiotics thereby facilitating the rapid measurement of the concentrations of all clinically important aminoglycosides in serum.

  8. Substrate specificity of fucosyltransferase purified from human parotid saliva.

    PubMed

    Tamagawa, H; Iwakura, K; Amano, A; Shizukuishi, S; Tsunemitsu, A

    1987-03-01

    The purified fucosyltransferase from human parotid saliva was shown to transfer fucose from GDP-fucose onto the oligosaccharide chains containing the Gal beta 1----3GlcNAc or Gal beta 1----4GlcNAc/Glc sequences. Competition studies between asialotransferrin and either lacto-N-fucopentaose 1 or 2'-fucosyllactose provided evidence that both the substrates competed for a common enzyme active site. These results suggest that the fucosyltransferase activities for the three acceptors may be catalyzed by the same enzyme.

  9. Magnetism for understanding catalyst analysis of purified carbon nanotubes

    NASA Astrophysics Data System (ADS)

    Bellouard, Christine; Mercier, Guillaume; Cahen, Sébastien; Ghanbaja, Jaafar; Medjahdi, Ghouti; Gleize, Jérôme; Lamura, Gianrico; Hérold, Claire; Vigolo, Brigitte

    2016-08-01

    The precise quantification of catalyst residues in purified carbon nanotubes is often a major issue in view of any fundamental and/or applicative studies. More importantly, since the best CNTs are successfully grown with magnetic catalysts, their quantification becomes strictly necessary to better understand intrinsic properties of CNT. For these reasons, we have deeply analyzed the catalyst content remained in nickel-yttrium arc-discharge single walled carbon nanotubes purified by both a chlorine-gas phase and a standard acid-based treatment. The study focuses on Ni analysis which has been investigated by transmission electron microscopy, X-ray diffraction, thermogravimetry analysis, and magnetic measurements. In the case of the acid-based treatment, all quantifications result in a decrease of the nanocrystallized Ni by a factor of two. In the case of the halogen gas treatment, analysis and quantification of Ni content is less straightforward: a huge difference appears between X-ray diffraction and thermogravimetry results. Thanks to magnetic measurements, this disagreement is explained by the presence of Ni2+ ions, belonging to NiCl2 formed during the Cl-based purification process. In particular, NiCl2 compound appears under different magnetic/crystalline phases: paramagnetic or diamagnetic, or well intercalated in between carbon sheets with an ordered magnetic phase at low temperature.

  10. PURIFY: a new approach to radio-interferometric imaging

    NASA Astrophysics Data System (ADS)

    Carrillo, R. E.; McEwen, J. D.; Wiaux, Y.

    2014-04-01

    In a recent paper series, the authors have promoted convex optimization algorithms for radio-interferometric imaging in the framework of compressed sensing, which leverages sparsity regularization priors for the associated inverse problem and defines a minimization problem for image reconstruction. This approach was shown, in theory and through simulations in a simple discrete visibility setting, to have the potential to outperform significantly CLEAN and its evolutions. In this work, we leverage the versatility of convex optimization in solving minimization problems to both handle realistic continuous visibilities and offer a highly parallelizable structure paving the way to significant acceleration of the reconstruction and high-dimensional data scalability. The new algorithmic structure promoted relies on the simultaneous-direction method of multipliers (SDMM) and contrasts with the current major-minor cycle structure of CLEAN and its evolutions, which in particular cannot handle the state-of-the-art minimization problems under consideration where neither the regularization term nor the data term are differentiable functions. We release a beta version of an SDMM-based imaging software written in C and dubbed PURIFY (http://basp-group.github.io/purify/) that handles various sparsity priors, including our recent average sparsity approach sparsity averaging reweighted analysis (SARA). We evaluate the performance of different priors through simulations in the continuous visibility setting, confirming the superiority of SARA.

  11. Synthesis of gold and silver nanoparticles using purified URAK.

    PubMed

    Deepak, Venkataraman; Umamaheshwaran, Paneer Selvam; Guhan, Kandasamy; Nanthini, Raja Amrisa; Krithiga, Bhaskar; Jaithoon, Nagoor Meeran Hasika; Gurunathan, Sangiliyandi

    2011-09-01

    This study aims at developing a new eco-friendly process for the synthesis of silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs) using purified URAK. URAK is a fibrinolytic enzyme produced by Bacillus cereus NK1. The enzyme was purified and used for the synthesis of AuNPs and AgNPs. The enzyme produced AgNPs when incubated with 1 mM AgNO3 for 24 h and AuNPs when incubated with 1 mM HAuCl4 for 60 h. But when NaOH was added, the synthesis was rapid and occurred within 5 min for AgNPs and 12 h for AuNPs. The synthesized nanoparticles were characterized by a peak at 440 nm and 550 nm in the UV-visible spectrum. TEM analysis showed that AgNPs of the size 60 nm and AuNPs of size 20 nm were synthesized. XRD confirmed the crystalline nature of the nanoparticles and AFM showed the morphology of the nanoparticle to be spherical. FT-IR showed that protein was responsible for the synthesis of the nanoparticles. This process is highly simple, versatile and produces AgNPs and AuNPs in environmental friendly manner. Moreover, the synthesized nanoparticles were found to contain immobilized enzyme. Also, URAK was tested on RAW 264.7 macrophage cell line and was found to be non-cytotoxic until 100 μg/ml.

  12. Monarch larvae sensitivity to Bacillus thuringiensis- purified proteins and pollen.

    PubMed

    Hellmich, R L; Siegfried, B D; Sears, M K; Stanley-Horn, D E; Daniels, M J; Mattila, H R; Spencer, T; Bidne, K G; Lewis, L C

    2001-10-09

    Laboratory tests were conducted to establish the relative toxicity of Bacillus thuringiensis (Bt) toxins and pollen from Bt corn to monarch larvae. Toxins tested included Cry1Ab, Cry1Ac, Cry9C, and Cry1F. Three methods were used: (i) purified toxins incorporated into artificial diet, (ii) pollen collected from Bt corn hybrids applied directly to milkweed leaf discs, and (iii) Bt pollen contaminated with corn tassel material applied directly to milkweed leaf discs. Bioassays of purified Bt toxins indicate that Cry9C and Cry1F proteins are relatively nontoxic to monarch first instars, whereas first instars are sensitive to Cry1Ab and Cry1Ac proteins. Older instars were 12 to 23 times less susceptible to Cry1Ab toxin compared with first instars. Pollen bioassays suggest that pollen contaminants, an artifact of pollen processing, can dramatically influence larval survival and weight gains and produce spurious results. The only transgenic corn pollen that consistently affected monarch larvae was from Cry1Ab event 176 hybrids, currently <2% corn planted and for which re-registration has not been applied. Results from the other types of Bt corn suggest that pollen from the Cry1Ab (events Bt11 and Mon810) and Cry1F, and experimental Cry9C hybrids, will have no acute effects on monarch butterfly larvae in field settings.

  13. Tea Derived Galloylated Polyphenols Cross-Link Purified Gastrointestinal Mucins

    PubMed Central

    Georgiades, Pantelis; Pudney, Paul D. A.; Rogers, Sarah; Thornton, David J.; Waigh, Thomas A.

    2014-01-01

    Polyphenols derived from tea are thought to be important for human health. We show using a combination of particle tracking microrheology and small-angle neutron scattering that polyphenols acts as cross-linkers for purified gastrointestinal mucin, derived from the stomach and the duodenum. Both naturally derived purified polyphenols, and green and black tea extracts are shown to act as cross-linkers. The main active cross-linking component is found to be the galloylated forms of catechins. The viscosity, elasticity and relaxation time of the mucin solutions experience an order of magnitude change in value upon addition of the polyphenol cross-linkers. Similarly small-angle neutron scattering experiments demonstrate a sol-gel transition with the addition of polyphenols, with a large increase in the scattering at low angles, which is attributed to the formation of large scale (>10 nm) heterogeneities during gelation. Cross-linking of mucins by polyphenols is thus expected to have an impact on the physicochemical environment of both the stomach and duodenum; polyphenols are expected to modulate the barrier properties of mucus, nutrient absorption through mucus and the viscoelastic microenvironments of intestinal bacteria. PMID:25162539

  14. Activation of purified calcium channels by stoichiometric protein phosphorylation

    SciTech Connect

    Nunoki, K.; Florio, V.; Catterall, W.A. )

    1989-09-01

    Purified dihydropyridine-sensitive calcium channels from rabbit skeletal muscle were reconstituted into phosphatidylcholine vesicles to evaluate the effect of phosphorylation by cyclic AMP-dependent protein kinase (PK-A) on their function. Both the rate and extent of {sup 45}Ca{sup 2+} uptake into vesicles containing reconstituted calcium channels were increased severalfold after incubation with ATP and PK-A. The degree of stimulation of {sup 45}Ca{sup 2+} uptake was linearly proportional to the extent of phosphorylation of the alpha 1 and beta subunits of the calcium channel up to a stoichiometry of approximately 1 mol of phosphate incorporated into each subunit. The calcium channels activated by phosphorylation were determined to be incorporated into the reconstituted vesicles in the inside-out orientation and were completely inhibited by low concentrations of dihydropyridines, phenylalkylamines, Cd{sup 2+}, Ni{sup 2+}, and Mg{sup 2+}. The results demonstrate a direct relationship between PK-A-catalyzed phosphorylation of the alpha 1 and beta subunits of the purified calcium channel and activation of the ion conductance activity of the dihydropyridine-sensitive calcium channels.

  15. Life cycle assessment comparison of photocatalytic coating and air purifier.

    PubMed

    Tichá, Marie; Žilka, Miroslav; Stieberová, Barbora; Freiberg, František

    2016-07-01

    This article presents a comparison of 2 very different options for removal of undesirable microorganisms and airborne pollutants from the indoor environment of hospitals, schools, homes, and other enclosed spaces using air purifiers and photocatalytic coatings based on nano titanium dioxide (TiO2 ). Both products were assessed by life cycle assessment (LCA) methodology from cradle-to-grave. The assessment also includes comparison of 2 different nano TiO2 production technologies, one by continuous hydrothermal synthesis and the other by a sulfate process. Results of the study showed a relatively large contribution of photocatalytic coatings to reducing the effects of selected indices in comparison with an air purifier, regardless of which nano TiO2 production method is used. Although the impacts of the sulfate process are significantly lower compared to those of hydrothermal synthesis when viewed in terms of production alone, taken in the context of the entire product life cycle, the net difference becomes less significant. The study has been elaborated within the Sustainable Hydrothermal Manufacturing of Nanomaterials (SHYMAN) project, which aims to develop competitive and sustainable continuous nanoparticle (NP) production technology based on supercritical hydrothermal synthesis. Integr Environ Assess Manag 2016;12:478-485. © 2016 SETAC.

  16. Foam mat drying of Tommy Atkins mango: Effects of air temperature and concentrations of soy lecithin and carboxymethylcellulose on phenolic composition, mangiferin, and antioxidant capacity.

    PubMed

    Lobo, Francine Albernaz; Nascimento, Manuela Abreu; Domingues, Josiane Roberto; Falcão, Deborah Quintanilha; Hernanz, Dolores; Heredia, Francisco J; de Lima Araujo, Kátia Gomes

    2017-04-15

    In this study, foam mat drying was applied to Tommy Atkins mango. Using a multifactorial design, the effect of soy lecithin (L) and carboxymethylcellulose (CMC) used as foam stabilizers (0-1.50g/100g), as well as temperature (T) (53-87°C), on phenolic content and antioxidant capacity of mango were evaluated. Mango pulp contains antioxidant, such as mangiferin, that can be utilized in foods to enhance their functional properties. Our results indicated that L and T had negative effects (p<0.05) on the phenolic content and antioxidant capacity, whereas CMC had a positive effect (p<0.05). Increasing the total amount of phenolic compounds present in dried mango contributed to the higher antioxidant capacity after the drying process. This study concluded that a drying T of 80°C, and a concentration of 0.30g/100g of CMC and L are optimal for increased retention of phenolic compounds and antioxidant capacity.

  17. Targeting of metastasis-promoting tumor-associated fibroblasts and modulation of pancreatic tumor-associated stroma with a carboxymethylcellulose-docetaxel nanoparticle.

    PubMed

    Ernsting, Mark J; Hoang, Bryan; Lohse, Ines; Undzys, Elijus; Cao, Pinjiang; Do, Trevor; Gill, Bethany; Pintilie, Melania; Hedley, David; Li, Shyh-Dar

    2015-05-28

    Pancreatic ductal adenocarcinomas are characterized by the desmoplastic reaction, a dense fibrous stroma that has been shown to be supportive of tumor cell growth, invasion, and metastasis, and has been associated with resistance to chemotherapy and reduced patient survival. Here, we investigated targeted depletion of stroma for pancreatic cancer therapy via taxane nanoparticles. Cellax-DTX polymer is a conjugate of docetaxel (DTX), polyethylene glycol (PEG), and acetylated carboxymethylcellulose, a construct which condenses into well-defined 120nm particles in an aqueous solution, and is suitable for intravenous injection. We examined Cellax-DTX treatment effects in highly stromal primary patient-derived pancreatic cancer xenografts and in a metastatic PAN02 mouse model of pancreatic cancer, focusing on specific cellular interactions in the stroma, pancreatic tumor growth and metastasis. Greater than 90% of Cellax-DTX particles accumulate in smooth muscle actin (SMA) positive cancer-associated fibroblasts which results in long-term depletion of this stromal cell population, an effect not observed with Nab-paclitaxel (Nab-PTX). The reduction in stromal density leads to a >10-fold increase in tumor perfusion, reduced tumor weight and a reduction in metastasis. Consentingly, Cellax-DTX treatment increased survival when compared to treatment with gemcitabine or Nab-PTX in a metastatic PAN02 mouse model. Cellax-DTX nanoparticles interact with the tumor-associated stroma, selectively interacting with and depleting SMA positive cells and macrophage, effects of which are associated with significant changes in tumor progression and metastasis.

  18. The influence of carboxymethylcellulose (CMC) on the reactivity of Fe NPs toward decabrominated diphenyl ether: The Ni doping, temperature, pH, and anion effects.

    PubMed

    Tso, Chih-Ping; Shih, Yang-Hsin

    2017-01-15

    Polybrominated diphenyl ethers (PBDEs) are commonly used brominated flame retardants in many products. They have accumulated in the environment and become widely dispersed. In this study, carboxymethylcellulose (CMC) was applied to modify nanoscale zerovalent iron (NZVI) and bimetallic Ni/Fe nanoparticles (NPs) to prevent NP aggregation. In this study the removal kinetics of the decabrominated diphenyl ethers (DBDE) with CMC-stabilized Fe NPs were evaluated. CMC-stabilized Ni/Fe NPs with an average size of 86.7nm contained metallic Fe(0) and reduced Ni. The colloidal stability decreased with a decrease in pH, which was further accompanied by a change in the removal rate of DBDE. Our results showed that anions do not change the removal rates of DBDE, with the exception of 10mM NO3(-), which induced the formation of Fe (hydro)oxides on the Fe NP surface, which could further coagulate with DBDE. This study provides important information for our understanding of the influence of CMC coatings on the reactivity of Fe NPs. Because CMC coatings prevent the passivation of Fe in the presence of anions, CMC-coated Fe NPs show potential for the in-situ remediation of PBDEs in the environment.

  19. CHARACTERIZATION OF THE PURIFIED HYALURONAN SYNTHASE FROM STREPTOCOCCUS EQUISIMILIS*

    PubMed Central

    Tlapak-Simmons, Valarie L.; Baron, Christina A.; Weigel, Paul H.

    2006-01-01

    Hyaluronan synthase (HAS) utilizes UDP-GlcUA and UDP-GlcNAc in the presence of Mg2+ to form the GAG hyaluronan (HA). The purified HAS from Streptococcus equisimilis (seHAS) shows high fidelity in that it only polymerizes the native substrates, UDP-GlcNAc and UDP-GlcUA. However, other uridinyl nucleotides and UDP-sugars inhibited enzyme activity, including UDP-GalNAc, UDP-Glc, UDP-Gal, UDP-GalUA, UMP, UDP and UTP. Purified seHAS was ~40% more active in 25 mM, compared to 50 mM, PO4 in the presence of either 50 mM NaCl or KCl, and displayed a slight preference for KCl over NaCl. The pH profile was surprisingly broad, with an effective range of pH 6.5–11.5 and the optimum between pH 9 and 10. SeHAS displayed two apparent pKa values at pH 6.6 and 11.8. As the pH was increased from ~6.5, both Km and Vmax increased until pH ~10.5, above which the kinetic constants gradually declined. Nonetheless, the overall catalytic constant (120/sec) was essentially unchanged from pH 6.5 to pH 10.5. The enzyme is temperature labile, but more stable in the presence of substrate and cardiolipin. Purified seHAS requires exogenous cardiolipin for activity and is very sensitive to the fatty acyl composition of the phospholipid. The enzyme was inactive or highly activated by synthetic cardiolipins containing, respectively, C14:0 or C18:1(Δ9) fatty acids. The apparent Ea for HA synthesis is 40 kJ (9.5 kcal/mol) disaccharide. Increasing the viscosity by increasing concentrations of PEG, ethylene glycol, glycerol, or sucrose inhibited seHAS activity. For PEGs, the extent of inhibition was proportional to their molecular mass. PEGs with average masses of 2.7, 11.7, and 20 Kg/mol caused 50% inhibition of Vmax at 21, 6.5, and 3.5 mM, respectively. The apparent Ki values for ethylene glycol, glycerol, and sucrose were, respectively, 4.5, 3.3 and 1.2 mM. PMID:15248781

  20. 42 CFR 84.170 - Non-powered air-purifying particulate respirators; description.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... that contain adequate oxygen to support life. (b) Non-powered air-purifying particulate respirators are... includes oil-based liquid particulates. (c) Non-powered air-purifying particulate respirators...

  1. Method and apparatus for removing residual hydrogen from a purified gas

    SciTech Connect

    Briesacher, J.L.; Applegarth, C.H.; Lorimer, D.H.

    1993-08-24

    A method is described for removing residual hydrogen from a purified gas comprising the steps of: (a) heating an impure gas; (b) contacting the heated impure gas with an impurity sorbing material to produce a purified gas having trace amounts of residual hydrogen; (c) cooling the purified gas to a temperature less than about 100 C; and (d) contacting the cooled purified gas with a hydrogen sorbing material to at least partially remove said residual hydrogen.

  2. Binding of rabies virus to purified Torpedo acetylcholine receptor.

    PubMed

    Lentz, T L; Benson, R J; Klimowicz, D; Wilson, P T; Hawrot, E

    1986-12-01

    The binding of 125I- and 35S-labeled rabies virus (CVS strain) to affinity-purified acetylcholine receptor from Torpedo electric organ was demonstrated. The binding of rabies virus to the acetylcholine receptor increased with increasing receptor concentration, was dependent on the pH of the incubation medium, and was saturable with increasing virus concentration. Binding of radioactively labeled virus was effectively competed by unlabeled homologous virus particles. Binding of 35S-labeled rabies virus to the AChR was inhibited up to 50% by alpha-bungarotoxin and up to 30% by (+)-tubocurarine but was not affected by atropine. These results demonstrate direct binding of rabies virus to a well-defined neurotransmitter receptor, namely the acetylcholine receptor and indicate that at least a portion of the virus interaction occurs near the acetylcholine binding site on the receptor. These findings support the hypothesis that the acetylcholine receptor may serve as a rabies virus receptor in vivo.

  3. X-ray diffraction study of highly purified human ceruloplasmin

    SciTech Connect

    Samygina, V. R.; Sokolov, A. V.; Pulina, M. O.; Bartunik, H. D.; Vasil'ev, V. B.

    2008-07-15

    The three-dimensional structure of ceruloplasmin (CP) with unoccupied labile metal-binding sites and the structure of CP containing Ni{sup 2+} in the labile sites were solved for the first time at 2.6 and 2.95 A resolution, respectively. Crystallization was performed with the use of storage-stable CP, which was prepared in the presence of proteinase inhibitors and purified from (pre)proteinases. Ceruloplasmin with Ni{sup 2+} crystallized in the orthorhombic space group, which had been earlier unknown for CP. Ceruloplasmin with the unoccupied labile sites crystallized in the trigonal crystal form. The differences in intermolecular contacts observed in the trigonal and orthorhombic crystal structures of CP are considered. The conformational changes attendant upon Ni{sup 2+} binding are described. It was suggested that the labile sites are multifunctional and can both bind metal ions potentially toxic to organisms and be involved in electron transfer from substrates to the active site.

  4. The Use of Detergents to Purify Membrane Proteins.

    PubMed

    Orwick-Rydmark, Marcella; Arnold, Thomas; Linke, Dirk

    2016-04-01

    Extraction of membrane proteins from biological membranes is usually accomplished with the help of detergents. This unit describes the use of detergents to solubilize and purify membrane proteins. The chemical and physical properties of the different classes of detergents typically used with biological samples are discussed. A separate section addresses the compatibility of detergents with applications downstream of the membrane protein purification process, such as optical spectroscopy, mass spectrometry, protein crystallography, biomolecular NMR, or electron microscopy. A brief summary of alternative membrane protein solubilizing and stabilizing systems is also included. Protocols in this unit include the isolation and solubilization of biological membranes and phase separation; support protocols for detergent removal, detergent exchange, and the determination of critical micelle concentration using different methods are also included.

  5. X-ray diffraction study of highly purified human ceruloplasmin

    NASA Astrophysics Data System (ADS)

    Samygina, V. R.; Sokolov, A. V.; Pulina, M. O.; Bartunik, H. D.; Vasil'Ev, V. B.

    2008-07-01

    The three-dimensional structure of ceruloplasmin (CP) with unoccupied labile metal-binding sites and the structure of CP containing Ni2+ in the labile sites were solved for the first time at 2.6 and 2.95 Å resolution, respectively. Crystallization was performed with the use of storage-stable CP, which was prepared in the presence of proteinase inhibitors and purified from (pre)proteinases. Ceruloplasmin with Ni2+ crystallized in the orthorhombic space group, which had been earlier unknown for CP. Ceruloplasmin with the unoccupied labile sites crystallized in the trigonal crystal form. The differences in intermolecular contacts observed in the trigonal and orthorhombic crystal structures of CP are considered. The conformational changes attendant upon Ni2+ binding are described. It was suggested that the labile sites are multifunctional and can both bind metal ions potentially toxic to organisms and be involved in electron transfer from substrates to the active site.

  6. Purifying Greenberger-Horne-Zeilinger states using degenerate quantum codes

    NASA Astrophysics Data System (ADS)

    Ho, K. H.; Chau, H. F.

    2008-10-01

    Degenerate quantum codes are codes that do not reveal the complete error syndrome. Their ability to conceal the complete error syndrome makes them powerful resources in certain quantum-information processing tasks. In particular, the most error-tolerant way to purify depolarized Bell states using one-way communication known to date involves degenerate quantum codes. Here we study three closely related purification schemes for depolarized Greenberger-Horne-Zeilinger states shared among m⩾3 players by means of degenerate quantum codes and one-way classical communications. We find that our schemes tolerate more noise than all other one-way schemes known to date, further demonstrating the effectiveness of degenerate quantum codes in quantum-information processing.

  7. DNA helicase activity in purified human RECQL4 protein.

    PubMed

    Suzuki, Takahiro; Kohno, Toshiyuki; Ishimi, Yukio

    2009-09-01

    Human RECQL4 protein was expressed in insect cells using a baculovirus protein expression system and it was purified to near homogeneity. The protein sedimented at a position between catalase (230 kDa) and ferritin (440 kDa) in glycerol gradient centrifugation, suggesting that it forms homo-multimers. Activity to displace annealed 17-mer oligonucleotide in the presence of ATP was co-sedimented with hRECQL4 protein. In ion-exchange chromatography, both DNA helicase activity and single-stranded DNA-dependent ATPase activity were co-eluted with hRECQL4 protein. The requirements of ATP and Mg for the helicase activity were different from those for the ATPase activity. The data suggest that the helicase migrates on single-stranded DNA in a 3'-5' direction. These results suggest that the hRECQL4 protein exhibits DNA helicase activity.

  8. Drosophilia spectrin. I. Characterization of the purified protein.

    PubMed

    Dubreuil, R; Byers, T J; Branton, D; Goldstein, L S; Kiehart, D P

    1987-11-01

    We purified a protein from Drosophila S3 tissue culture cells that has many of the diagnostic features of spectrin from vertebrate organisms: (a) The protein consists of two equimolar subunits (Mr = 234 and 226 kD) that can be reversibly cross-linked into a complex composed of equal amounts of the two subunits. (b) Electron microscopy of the native molecule reveals two intertwined, elongated strands with a contour length of 180 nm. (c) Antibodies directed against vertebrate spectrin react with the Drosophila protein and, similarly, antibodies to the Drosophila protein react with vertebrate spectrins. One monoclonal antibody has been found to react with both of the Drosophila subunits and with both subunits of vertebrate brain spectrin. (d) The Drosophila protein exhibits both actin-binding and calcium-dependent calmodulin-binding activities. Based on the above criteria, this protein appears to be a bona fide member of the spectrin family of proteins.

  9. Using ion exchange chromatography to purify a recombinantly expressed protein.

    PubMed

    Duong-Ly, Krisna C; Gabelli, Sandra B

    2014-01-01

    Ion exchange chromatography (IEX) separates molecules by their surface charge, a property that can vary vastly between different proteins. There are two types of IEX, cation exhange and anion exchange chromatography. The protocol that follows was designed by the authors for anion exchange chromatography of a recombinantly expressed protein having a pI of 4.9 and containing two cysteine residues and one tryptophan residue, using an FPLC system. Prior to anion exchange, the protein had been salted out using ammonium sulfate precipitation and partially purified via hydrophobic interaction chromatography (see Salting out of proteins using ammonium sulfate precipitation and Use and Application of Hydrophobic Interaction Chromatography for Protein Purification). Slight modifications to this protocol may be made to accommodate both the protein of interest and the availability of equipment.

  10. Measurement of Ozone Emission and Particle Removal Rates from Portable Air Purifiers

    ERIC Educational Resources Information Center

    Mang, Stephen A.; Walser, Maggie L.; Nizkorodov, Sergey A.; Laux, John M.

    2009-01-01

    Portable air purifiers are popular consumer items, especially in areas with poor air quality. Unfortunately, most users of these air purifiers have minimal understanding of the factors affecting their efficiency in typical indoor settings. Emission of the air pollutant ozone (O[subscript 3]) by certain air purifiers is of particular concern. In an…

  11. Characterization of purified New Delhi metallo-β-lactamase-1.

    PubMed

    Thomas, Pei W; Zheng, Min; Wu, Shanshan; Guo, Hua; Liu, Dali; Xu, Dingguo; Fast, Walter

    2011-11-22

    New Delhi metallo-β-lactmase-1 (NDM-1) has recently emerged as a global threat because of its ability to confer resistance to almost all clinically used β-lactam antibiotics, its presence within an easily transmissible plasmid bearing a number of other antibiotic resistance determinants, its carriage in a variety of enterobacteria, and its presence in both nosocomial and community-acquired infections. To improve our understanding of the molecular basis of this threat, NDM-1 was purified and characterized. Recombinant NDM-1 bearing its native leader sequence was expressed in Escherichia coli BL21 cells. The major processed form found to be released into culture media contains a 35-residue truncation at the N-terminus. This form of NDM-1 is monomeric and can be purified with 1.8 or 1.0 equiv of zinc ion, depending on the experimental conditions. Treatment of dizinc NDM-1 with EDTA results in complete removal of both zinc ions, but the relatively weaker chelator PAR chelates only 1 equiv of zinc ion from folded protein but 1.9 equiv of zinc ion from denatured protein, indicating different affinities for each metal binding site. UV-vis spectroscopy of the dicobalt metalloform along with molecular dynamics simulations of the dizinc metallo form indicates that the dinuclear metal cluster at the active site of NDM-1 is similar in structure to other class B1 metallo-β-lactamases. Supplementation of excess zinc ions to monozinc NDM-1 has differential effects on enzyme activity with respect to three different classes of β-lactam substrates tested, penems, cephems, and carbapenems, and likely reflects dissimilar contributions of the second equivalent of metal ion to the catalysis of the hydrolysis of these substrates. Fits to these concentration dependencies are used to approximate the K(d) value of the more weakly bound zinc ion (2 μM). NDM-1 achieved maximal activity with all substrates tested when supplemented with approximately 10 μM ZnSO(4), displaying k

  12. Widespread purifying selection on RNA structure in mammals.

    PubMed

    Smith, Martin A; Gesell, Tanja; Stadler, Peter F; Mattick, John S

    2013-09-01

    Evolutionarily conserved RNA secondary structures are a robust indicator of purifying selection and, consequently, molecular function. Evaluating their genome-wide occurrence through comparative genomics has consistently been plagued by high false-positive rates and divergent predictions. We present a novel benchmarking pipeline aimed at calibrating the precision of genome-wide scans for consensus RNA structure prediction. The benchmarking data obtained from two refined structure prediction algorithms, RNAz and SISSIz, were then analyzed to fine-tune the parameters of an optimized workflow for genomic sliding window screens. When applied to consistency-based multiple genome alignments of 35 mammals, our approach confidently identifies >4 million evolutionarily constrained RNA structures using a conservative sensitivity threshold that entails historically low false discovery rates for such analyses (5-22%). These predictions comprise 13.6% of the human genome, 88% of which fall outside any known sequence-constrained element, suggesting that a large proportion of the mammalian genome is functional. As an example, our findings identify both known and novel conserved RNA structure motifs in the long noncoding RNA MALAT1. This study provides an extensive set of functional transcriptomic annotations that will assist researchers in uncovering the precise mechanisms underlying the developmental ontologies of higher eukaryotes.

  13. Widespread purifying selection on RNA structure in mammals

    PubMed Central

    Smith, Martin A.; Gesell, Tanja; Stadler, Peter F.; Mattick, John S.

    2013-01-01

    Evolutionarily conserved RNA secondary structures are a robust indicator of purifying selection and, consequently, molecular function. Evaluating their genome-wide occurrence through comparative genomics has consistently been plagued by high false-positive rates and divergent predictions. We present a novel benchmarking pipeline aimed at calibrating the precision of genome-wide scans for consensus RNA structure prediction. The benchmarking data obtained from two refined structure prediction algorithms, RNAz and SISSIz, were then analyzed to fine-tune the parameters of an optimized workflow for genomic sliding window screens. When applied to consistency-based multiple genome alignments of 35 mammals, our approach confidently identifies >4 million evolutionarily constrained RNA structures using a conservative sensitivity threshold that entails historically low false discovery rates for such analyses (5–22%). These predictions comprise 13.6% of the human genome, 88% of which fall outside any known sequence-constrained element, suggesting that a large proportion of the mammalian genome is functional. As an example, our findings identify both known and novel conserved RNA structure motifs in the long noncoding RNA MALAT1. This study provides an extensive set of functional transcriptomic annotations that will assist researchers in uncovering the precise mechanisms underlying the developmental ontologies of higher eukaryotes. PMID:23847102

  14. Permeation of Calcium through Purified Connexin 26 Hemichannels*

    PubMed Central

    Fiori, Mariana C.; Figueroa, Vania; Zoghbi, Maria E.; Saéz, Juan C.; Reuss, Luis; Altenberg, Guillermo A.

    2012-01-01

    Gap junction channels communicate the cytoplasms of two cells and are formed by head to head association of two hemichannels, one from each of the cells. Gap junction channels and hemichannels are permeable to ions and hydrophilic molecules of up to Mr 1,000, including second messengers and metabolites. Intercellular Ca2+ signaling can occur by movement of a number of second messengers, including Ca2+, through gap junction channels, or by a paracrine pathway that involves activation of purinergic receptors in neighboring cells following ATP release through hemichannels. Understanding Ca2+ permeation through Cx26 hemichannels is important to assess the role of gap junction channels and hemichannels in health and disease. In this context, it is possible that increased Ca2+ influx through hemichannels under ischemic conditions contributes to cell damage. Previous studies suggest Ca2+ permeation through hemichannels, based on indirect arguments. Here, we demonstrate for the first time hemichannel permeability to Ca2+ by measuring Ca2+ transport through purified Cx26 hemichannels reconstituted in liposomes. We trapped the low affinity Ca2+-sensitive fluorescent probe Fluo-5N into the liposomes and followed the increases in intraliposomal [Ca2+] in response to an imposed [Ca2+] gradient. We show that Ca2+ does move through Cx26 hemichannels and that the permeability of the hemichannels to Ca2+ is high, similar to that for Na+. We suggest that hemichannels can be a significant pathway for Ca2+ influx into cells under conditions such as ischemia. PMID:23048025

  15. Differentiation of purified astrocytes in a chemically defined medium

    SciTech Connect

    Morrison, R.S.; de Vellis, J.

    1981-01-01

    Homogeneous cultures of astrocytes and oligodendrocytes provide an excellent model system for studying the regulation of glial structure and function. Recently, a chemically defined (CD) medium was developed for purified cultures of astrocytes, thus eliminating the requirement for serum and providing a controlled system for the study of astroglial properties. Due to the widespread use of astrocyte cultures and the potential benefits to be gained from using a defined medium, astrocyte cultures raised in CD medium were analyzed for purity as well as morphological and biochemical properties. Purity was assessed using immunocytochemical staining for glial fibrillary acidic protein (GFAP) and fibronectin. Astrocytes raised in CD medium are 95% pure using the expression of GFAP as a criterion. Fewer than 1% of the cells in CD medium stained positive for fibronectin eliminating the possibility that CD medium is selective for meningeal or endothelial cells. Astrocytes raised in CD medium exhibit a striking degree of morphological differentiation as seen in scanning electron micrographs. They also exhibit a high degree of biochemical differentiation illustrated by increases in the specific activity of S-100 protein and the induction of glutamine synthetase by glucocorticoids. A defined medium that supports the proliferation of rat astrocytes and enhances numerous morphological and biochemical properties should greatly facilitate the study of factors controlling glial proliferation and differentiation.

  16. Simple, fast, and efficient process for producing and purifying trehalulose.

    PubMed

    Wei, Yutuo; Liang, Jiayuan; Huang, Ying; Lei, Panxian; Du, Liqin; Huang, Ribo

    2013-06-01

    A new property of recombinant trehalose synthase (GTase) from Thermus thermophilus HB-8 (ATCC 27634) was found and described in this study. GTase can act on sucrose and catalyze trehalulose formation without isomaltose, isomaltulose, or isomelezitose, releasing small amounts of glucose and fructose as byproducts. Maximum trehalulose yield (approximately 81%) was obtained at an optimum temperature of 65°C and was independent of substrate concentration. A simple, fast, and efficient method of producing and purifying trehalulose is then described. In the first step, GTase catalyzed trehalulose formation using a 20% sucrose substrate. Miscellaneous sugars were then rapidly removed, while trehalulose was completely preserved by Saccharomyces cerevisiae cells. Finally, the cells were separated by centrifugation, and salt ions were removed by an ion-exchange resin, subsequently obtaining a high-purity trehalulose solution. A trehalulose recovery rate of over 95% was achieved using this process. This method has a simple process, fast separation efficiency, and low investment in production equipment, so greatly to improve production efficiency and reduce production costs.

  17. Aflatoxin detoxification by manganese peroxidase purified from Pleurotus ostreatus

    PubMed Central

    Yehia, Ramy Sayed

    2014-01-01

    Manganese peroxidase (MnP) was produced from white rot edible mushroom Pleurotus ostreatus on the culture filtrate. The enzyme was purified to homogeneity using (NH4)2SO4 precipitation, DEAE-Sepharose and Sephadex G-100 column chromatography. The final enzyme activity achieved 81 U mL−1, specific activity 78 U mg−1 with purification fold of 130 and recovery 1.2% of the crude enzyme. SDS-PAGE indicated that the pure enzyme have a molecular mass of approximately 42 kDa. The optimum pH was between 4–5 and the optimum temperature was 25 °C. The pure MnP activity was enhanced by Mn2+, Cu2+, Ca2+ and K+ and inhibited by Hg+2 and Cd+2. H2O2 at 5 mM enhanced MnP activity while at 10 mM inhibited it significantly. The MnP-cDNA encoding gene was sequenced and determined (GenBank accession no. AB698450.1). The MnP-cDNA was found to consist of 497 bp in an Open Reading Frame (ORF) encoding 165 amino acids. MnP from P. ostreatus could detoxify aflatoxin B1 (AFB1) depending on enzyme concentration and incubation period. The highest detoxification power (90%) was observed after 48 h incubation at 1.5 U mL−1 enzyme activities. PMID:24948923

  18. Bioavailability of purified subcellular metals to a marine fish.

    PubMed

    Guo, Feng; Yao, Jie; Wang, Wen-Xiong

    2013-09-01

    In the present study, the authors used a supply of naturally contaminated oysters to investigate how the subcellular metal distribution and the metal burden in prey affected the transfer of metals to a marine fish, the grunt Terapon jarbua. The oysters, Crassostrea hongkongensis, each with different contamination histories, were collected and separated into 3 subcellular fractions: 1) metal-rich granules, 2) cellular debris, and 3) a combined fraction of organelles, heat-denatured proteins, and metallothionein-like proteins, defined as the trophically available metal (TAM). These purified fractions showed a wide range of metal concentrations and were fed to the fish for a period of 7 d at a daily comparable feeding rate of 3% of fish body weight. After 7 d exposure, the newly absorbed metals were mainly distributed in the intestine and liver, indicating a significant tissue-specific trophic transfer, especially for Cd and Cu. The trophic transfer factors (TTFs) showed a sequence of cellular debris >TAM > metal-rich granules, suggesting the impact of subcellular distribution in prey on metal bioavailability. However, significant inverse relationships between the TTFs and the metal concentrations in diets were also found in the present study, especially for Cd and Zn. The subcellular metal compartmentalization might be less important than the metal concentration in prey influencing the trophic transfer. The authors' results have important implications for bioavailability and environmental assessment of dietary metals.

  19. [Micronized purified flavonoid fraction in treatment of pelvic varicose veins].

    PubMed

    Gavrilov, S G; Karalkin, A V; Moskalenko, E P; Beliaeva, E S; Ianina, A M; Kirienko, A I

    2012-01-01

    Presented herein are the results of studying efficacy of micronized purified flavonoid fraction (MPFF) in treatment of pelvic varicose veins (PVV) using reference ray-tracing methods of study. We examined a total of 85 patients with PVV. Of these, 65 subjects were found to have isolated dilatation of pelvic venous plexuses (study group), and 20 were diagnosed as having combined dilation of gonadal veins and venous plexuses of the pelvis (control group). Besides clinical examination, the patients were subjected to ultrasonographic angioscanning (USAS) and emission computed tomography (ECT) of pelvic veins before treatment and 2, 6, 12, 24, 36 and 60 months after the beginning of phlebotrophic therapy. Based on the findings of the clinical and instrumental studies, it was determined that MPFF was most efficient in patients with isolated dilatation of uterine and parametrial veins. In this group of patients, pelvic pain and other symptoms of the disease disappeared completely and the clinical effect persisted for a long time (up to 6-9 months). In the control group, venotonic therapy had a positive effect which was less pronounced as compared to the control group, and pelvic pain reappeared in the nearest time (up to 3 weeks) after withdrawal of MPFF.

  20. Functional nucleus pulposus-like matrix assembly by human mesenchymal stromal cells is directed by macromer concentration in photocrosslinked carboxymethylcellulose hydrogels.

    PubMed

    Gupta, Michelle S; Nicoll, Steven B

    2014-11-01

    Intervertebral disc (IVD) degeneration is associated with several pathophysiologic changes of the IVD, including dehydration of the nucleus pulposus (NP). Tissue engineering strategies may be used to restore both biological and mechanical function of the IVD following removal of NP tissue during surgical intervention. Recently, photocrosslinked carboxymethylcellulose (CMC) hydrogels were shown to support chondrogenic, NP-like extracellular matrix (ECM) elaboration by human mesenchymal stromal cells (hMSCs) when supplemented with TGF-β3; however, mechanical properties of these constructs did not reach native values. Fabrication parameters (i.e., composition, crosslinking density) can influence the bulk mechanical properties of hydrogel scaffolds, as well as cellular behavior and differentiation patterns. The objective of this study was to evaluate the influence of CMC macromer concentration (1.5, 2.5 and 3.5 % weight/volume) on bulk hydrogel properties and NP-like matrix elaboration by hMSCs. The lowest macromer concentration of 1.5 % exhibited the highest gene expression levels of aggrecan and collagen II at day 7, corresponding with the largest accumulation of glycosaminoglycans and collagen II by day 42. The ECM elaboration in the 1.5 % constructs was more homogeneously distributed compared to primarily pericellular localization in 3.5 % gels. The 1.5 % gels also displayed significant improvements in mechanical functionality by day 42 compared to earlier time points, which was not seen in the other groups. The effects of macromer concentration on matrix accumulation and organization are likely attributed to quantifiable differences in polymer crosslinking density and diffusive properties between the various hydrogel formulations. Taken together, these results demonstrate that macromer concentration of CMC hydrogels can direct hMSC matrix elaboration, such that a lower polymer concentration allows for greater NP-like ECM assembly and improvement of mechanical

  1. Efficacy and Safety of Sodium Hyaluronate with 1,4-Butanediol Diglycidyl Ether Compared to Sodium Carboxymethylcellulose in Preventing Adhesion Formation after Lumbar Discectomy

    PubMed Central

    Ji, Gyu Yeul; Oh, Chang Hyun; Moon, Byung Gwan; Yi, Seong; Han, In Bo; Heo, Dong Hwa; Kim, Ki-Tack; Kim, Keung Nyun

    2015-01-01

    Objective Epidural injection of hyaluronic acid may prevent adhesion formation after spine surgery, but the compounds used to stabilize hyaluronidase could interfere with its anti-adhesion effects. The present study was conducted as a clinical trial to evaluate the efficacy and safety of an experimental medical gel in preventing adhesion formation. Methods This study was designed as a multicenter, randomized, double-blind, and comparative controlled clinical trial with an observation period of 6 weeks. Subjects were randomly assigned into two groups: group A with sodium hyaluronate + 1,4-butanediol diglycidyl ether (BDDE) and group B with sodium hyaluronate + sodium carboxymethylcellulose (CMC). Visual analogue scale (VAS) of back and leg pain and the Oswestry disability index (ODI) and scar score ratings were assessed after surgery. Results Mean scar grade was 2.37±1.13 in group A and 2.75±0.97 in group B, a statistically significant difference (p=0.012). VAS of back and leg pain and ODI scores decreased significantly from baseline to 3 and 6 weeks postoperatively in both groups (p<0.001). However, VAS and ODI scores were not statistically different between groups A and B at baseline or at 3 and 6 weeks after operation (p>0.3). The number of adverse reactions related to the anti-adhesion gels was not statistically different (p=0.569), but subsequent analysis of nervous adverse reactions showed group B was superior with a statistically difference (p=0.027). Conclusion Sodium hyaluronate with BDDE demonstrated similar anti-adhesion properties to sodium hyaluronate with CMC. But, care should be used to nervous adverse reactions by using sodium hyaluronate with BDDE. PMID:26217381

  2. Access to small size distributions of nanoparticles by microwave-assisted synthesis. Formation of Ag nanoparticles in aqueous carboxymethylcellulose solutions in batch and continuous-flow reactors.

    PubMed

    Horikoshi, Satoshi; Abe, Hideki; Torigoe, Kanjiro; Abe, Masahiko; Serpone, Nick

    2010-08-01

    This article examines the effect(s) of the 2.45-GHz microwave (MW) radiation in the synthesis of silver nanoparticles in aqueous media by reduction of the diaminesilver(i) complex, [Ag(NH(3))(2)](+), with carboxymethylcellulose (CMC) in both batch-type and continuous-flow reactor systems with a particular emphasis on the characteristics of the microwaves in this process and the size distributions. This microwave thermally-assisted synthesis is compared to a conventional heating (CH) method, both requiring a reaction temperature of 100 degrees C to produce the nanoparticles, in both cases leading to the formation of silver colloids with different size distributions. Reduction of the diaminesilver(i) precursor complex, [Ag(NH(3))(2)](+), by CMC depended on the solution temperature. Cooling the reactor during the heating process driven with 390-Watt microwaves (MW-390W/Cool protocol) yielded silver nanoparticles with sizes spanning the range 1-2 nm. By contrast, the size distribution of Ag nanoparticles with 170-Watt microwaves (no cooling; MW-170W protocol) was in the range 1.4-3.6 nm (average size approximately 3 nm). The overall results suggest the potential for a scale-up process in the microwave-assisted synthesis of nanoparticles. Based on the present data, a flow-through microwave reactor system is herein proposed for the continuous production of silver nanoparticles. The novel flow reactor system (flow rate, 600 mL min(-1)) coupled to 1200-Watt microwave radiation generated silver nanoparticles with a size distribution 0.7-2.8 nm (average size ca. 1.5 nm).

  3. Application of in-line viscometer for in-process monitoring of microcrystalline cellulose-carboxymethylcellulose hydrogel formation during batch manufacturing.

    PubMed

    Pu, Yu; Chaudhry, Saeed; Parikh, Maider; Berry, Julianne

    2015-01-01

    Physical stability and consistent dose delivery of pharmaceutical suspension formulations comprised of microcrystalline cellulose (MCC) and sodium carboxymethylcellulose (NaCMC) hydrogels is dependent on their rheological properties. To obtain the desired rheological characteristics, good control of the hydrogel dispersion in water is required. The goal of this study was to evaluate whether the XL7-100 Process Viscometer could be employed as a process analytical technology (PAT) tool to monitor the dispersion process in real time during batch manufacturing. Using this instrument, viscosity profiles were measured during the hydrogel processing for a range of operating conditions. It was confirmed that viscosity obtained by the XL7-100 Process Viscometer in the off-line mode, could be linearly correlated to that of the conventional Brookfield viscometer. In addition, the XL7-100 Process Viscometer was able to detect variations in the hydrogel concentrations as well as process conditions in real time. Under fixed operating conditions, the dynamic viscosity profile showed low variability and good inter-batch reproducibility for a properly dispersed hydrogel. For a well-validated mixing process, an off-trend in-line viscosity reading may be indicative of batch failure or poor dispersion homogeneity. Therefore, the in-line viscometer can be used in manufacturing to monitor the batch to batch consistency. However, it is not proven to be able to characterize the real-time structure formation of the hydrogel. It is recommended that the in-line viscometer be used as a complimentary tool along with the off-line rheometer for both efficient and effective in-process quality control of the MCC & NaCMC hydrogel dispersion.

  4. Purified Shigella enterotoxin does not alter intestinal motility.

    PubMed Central

    Fernandez, A; Sninsky, C A; O'Brien, A D; Clench, M H; Mathias, J R

    1984-01-01

    A purified Shigella enterotoxin (pST) and a cell-free lysate with pST removed (CFL-pST) from the whole-cell lysate of Shigella dysenteriae 60 R were used to study their effect on the myoelectric activity and mucosal integrity of rabbit ileal segments. We have previously defined two myoelectric patterns: the migrating action potential complex and repetitive bursts of action potentials that occur in response to certain bacteria and their enterotoxins. The in vivo model consisted of isolated ileal segments in male New Zealand White rabbits. The segments were infused with sterile saline (1 ml/h), pST (2.4-micrograms injection), or CFL-pST (1 ml/h). Myoelectric activity in the segments exposed to pST was similar to that with the saline infusion, but CFL-pST induced significant alterations in myoelectric activity in the form of repetitive bursts of action potentials. The mucosa of the segments exposed to pST showed only mild inflammatory changes. In contrast, CFL-pST caused moderate to severe inflammatory changes with enterocyte necrosis. These studies show that pST, a known enterotoxin, did not alter myoelectric activity and had no significant effect on the integrity of ileal mucosa, as determined by light microscopy. CFL-pST caused both inflammation and tissue necrosis with significant alterations in motor activity. These studies suggest that S. dysenteriae 60 R produces a substance or substances other than pST that cause florid in vivo cytotoxicity and alter myoelectric activity. Images PMID:6363286

  5. Development of a Microwave Regenerative Sorbent-Based Hydrogen Purifier

    NASA Technical Reports Server (NTRS)

    Wheeler, Richard R., Jr.; Dewberry, Ross H.; McCurry, Bryan D.; Abney, Morgan B.; Greenwood, Zachary W.

    2016-01-01

    This paper describes the design and fabrication of a Microwave Regenerative Sorbent-based Hydrogen Purifier (MRSHP). This unique microwave powered technology was developed for the purification of a hydrogen stream produced by the Plasma Pyrolysis Assembly (PPA). The PPA is a hydrogen recovery (from methane) post processor for NASA's Sabatier-based carbon dioxide reduction process. Embodied in the Carbon dioxide Reduction Assembly (CRA), currently aboard the International Space Station (ISS), the Sabatier reaction employs hydrogen to catalytically recover oxygen, in the form of water, from respiratory carbon dioxide produced by the crew. This same approach is base-lined for future service in the Air Revitalization system on extended missions into deep space where resupply is not practical. Accordingly, manned exploration to Mars may only become feasible with further closure of the air loop as afforded by the greater hydrogen recovery permitted by the PPA with subsequent hydrogen purification. By utilizing the well-known high sorbate loading capacity of molecular sieve 13x, coupled with microwave dielectric heating phenomenon, MRSHP technology is employed as a regenerative filter for a contaminated hydrogen gas stream. By design, freshly regenerated molecular sieve 13x contained in the MRSHP will remove contaminants from the effluent of a 1-CM scale PPA for several hours prior to breakthrough. By reversing flow and pulling a relative vacuum the MRSHP prototype then uses 2.45 GHz microwave power, applied through a novel coaxial antenna array, to rapidly heat the sorbent bed and drive off the contaminants in a short duration vacuum/thermal contaminant desorption step. Finally, following rapid cooling via room temperature cold plates, the MRSHP is again ready to serve as a hydrogen filter.

  6. Correcting for Purifying Selection: An Improved Human Mitochondrial Molecular Clock

    PubMed Central

    Soares, Pedro; Ermini, Luca; Thomson, Noel; Mormina, Maru; Rito, Teresa; Röhl, Arne; Salas, Antonio; Oppenheimer, Stephen; Macaulay, Vincent; Richards, Martin B.

    2009-01-01

    There is currently no calibration available for the whole human mtDNA genome, incorporating both coding and control regions. Furthermore, as several authors have pointed out recently, linear molecular clocks that incorporate selectable characters are in any case problematic. We here confirm a modest effect of purifying selection on the mtDNA coding region and propose an improved molecular clock for dating human mtDNA, based on a worldwide phylogeny of > 2000 complete mtDNA genomes and calibrating against recent evidence for the divergence time of humans and chimpanzees. We focus on a time-dependent mutation rate based on the entire mtDNA genome and supported by a neutral clock based on synonymous mutations alone. We show that the corrected rate is further corroborated by archaeological dating for the settlement of the Canary Islands and Remote Oceania and also, given certain phylogeographic assumptions, by the timing of the first modern human settlement of Europe and resettlement after the Last Glacial Maximum. The corrected rate yields an age of modern human expansion in the Americas at ∼15 kya that—unlike the uncorrected clock—matches the archaeological evidence, but continues to indicate an out-of-Africa dispersal at around 55–70 kya, 5–20 ky before any clear archaeological record, suggesting the need for archaeological research efforts focusing on this time window. We also present improved rates for the mtDNA control region, and the first comprehensive estimates of positional mutation rates for human mtDNA, which are essential for defining mutation models in phylogenetic analyses. PMID:19500773

  7. Enzymatic characterization of recombinant nitrate reductase expressed and purified from Neurospora crassa.

    PubMed

    Ringel, Phillip; Probst, Corinna; Dammeyer, Thorben; Buchmeier, Sabine; Jänsch, Lothar; Wissing, Josef; Tinnefeld, Philip; Mendel, Ralf R; Jockusch, Brigitte M; Kruse, Tobias

    2015-07-01

    We established an expression and purification procedure for recombinant protein production in Neurospora crassa (N. crassa). This Strep-tag® based system was successfully used for purifying recombinant N. crassa nitrate reductase (NR), whose enzymatic activity was compared to recombinant N. crassa NR purified from Escherichia coli. The purity of the two different NR preparations was similar but NR purified from N. crassa showed a significantly higher nitrate turnover rate. Two phosphorylation sites were identified for NR purified from the endogenous expression system. We conclude that homologous expression of N. crassa NR yields a higher active enzyme and propose that NR phosphorylation causes enhanced enzymatic activity.

  8. Optical properties and ensemble characteristics of size purified Silicon nanocrystals

    NASA Astrophysics Data System (ADS)

    Miller, Joseph Bradley

    Nanotechnology is at the forefront of current scientific research and nanocrystals are being hailed as the 'artificial' atoms of the 21st century. Semiconducting silicon nanocrystals (SiNCs) are prime candidates for potential commercial applications because of silicon's already ubiquitous presence in the semiconductor industry, nontoxicity and abundance in nature. For realization of these potential applications, the properties and behavior of SiNCs need to be understood and enhanced. In this report, some of the main SiNC synthesis schemes are discussed, including those we are currently experimenting with to create our own SiNCs and the one utilized to create the SiNCs used in this study. The underlying physics that governs the unique behavior of SiNCs is then presented. The properties of the as-produced SiNCs are determined to depend strongly on surface passivation and environment. Size purification, an important aspect of nanomaterial utilization, was successfully performed on our SiNCs though density gradient ultracentrifugation. We demonstrate that the size-purified fractions exhibit an enhanced ability for colloidal self-assembly, with better aligned nanocrystal energy levels which promotes greater photostability in close-packed films and produces a slight increase in photoluminescence (PL) quantum yield. The qualities displayed by the fractions are exploited to form SiNC clusters that exhibit photostable PL. An analysis of SiNC cluster (from individual nanocrystals to collections of more than one thousand) blinking and PL shows an improvement in their PL emitting 'on' times. Pure SiNC films and SiNC-polymer nanocomposites are created and the dependence of their PL on temperature is measured. For such nanocomposites, the coupling between the 'coffee-ring' effect and liquid-liquid phase separation is also examined for ternary mixtures of solvent, polymer and semiconducting nanocrystal. We discover that with the right SiNC-polymer concentration and polymer

  9. Designing green oxidation catalysts for purifying environmental waters

    PubMed Central

    Ellis, W. Chadwick; Tran, Camly T.; Roy, Riddhi; Rusten, Marte; Fischer, Andreas; Ryabov, Alexander D.

    2010-01-01

    We describe the synthesis, characterization, aqueous behavior, and catalytic activity of a new generation of FeIII-TAML (TetraAmido Macrocycle Ligand) activators of peroxides (2), variants of (2d), which have been designed to be especially suitable for purifying water of recalcitrant oxidizable pollutants. Activation of H2O2 by 2 (kI) as a function of pH was analyzed via kinetic studies of Orange II bleaching. This was compared with the known behavior of the first generation of FeIII-TAMLs (1). Novel reactivity features impact the potential for oxidant activation for water purification by 2d and its aromatic ring substituted dinitro (2e) and tetrachloro (2f) derivatives. Thus, the maximum activity for 2e occurs at pH 9, the closest yet to the EPA guidelines for drinking water (6.5–8.5) allowing 2e to rapidly activate H2O2 at pH 7.7. In water, 2e has two axial water ligands with pKas of 8.4 and 10.0 (25 °C). The former is the lowest for all FeIII-TAMLs developed to date and is key to 2e’s exceptional catalytic activity in neutral and slightly basic solutions. Below pH 7 2d was found to be quite sensitive to demetalation in phosphate buffers. This was overcome by iterative design to give 2e (hydrolysis rate 2d > 100×2e). Mechanistic studies highlight 2e’s increased stability by establishing that to demetalate 2e at a comparable rate to which H2PO4− demetalates 2d, H3PO4 is required. A critical criterion for green catalysts for water purification is the avoidance of endocrine disruptors, which can impair aquatic life. FeIII-TAMLs do not alter transcription mediated by mammalian thyroid, androgen or estrogen hormone receptors, suggesting that 2 do not bind to the receptors and reducing concerns that the catalysts might have endocrine disrupting activity. PMID:20565079

  10. Hypolipidemic activities of xanthorrhizol purified from centrifugal TLC.

    PubMed

    Oon, Seok Fang; Nallappan, Meenakshii; Kassim, Nur Kartinee; Shohaimi, Shamarina; Sa'ariwijaya, Mohd Shazrul Fazry; Tee, Thiam Tsui; Cheah, Yew Hoong

    2016-09-23

    Hyperlipidemia is defined as the presence of either hypertriglyceridemia or hypercholesterolemia, which could cause atherosclerosis. Although hyperlipidemia can be treated by hypolipidemic drugs, they are limited due to lack of effectiveness and safety. Previous studies demonstrated that xanthorrhizol (XNT) isolated from Curcuma xanthorrhizza Roxb. reduced the levels of free fatty acid and triglyceride in vivo. However, its ability to inhibit cholesterol uptake in HT29 colon cells and adipogenesis in 3T3-L1 cells are yet to be reported. In this study, XNT purified from centrifugal TLC demonstrated 98.3% purity, indicating it could be an alternative purification method. The IC50 values of XNT were 30.81 ± 0.78 μg/mL in HT29 cells and 35.07 ± 0.24 μg/mL in 3T3-L1 adipocytes, respectively. Cholesterol uptake inhibition study using HT29 colon cells showed that XNT (15 μg/mL) significantly inhibited the fluorescent cholesterol analogue NBD uptake by up to 27 ± 3.1% relative to control. On the other hand, higher concentration of XNT (50 μg/mL) significantly suppressed the growth of 3T3-L1 adipocytes (5.9 ± 0.58%) compared to 3T3-L1 preadipocytes (81.31 ± 0.55%). XNT was found to impede adipogenesis of 3T3-L1 adipocytes in a dose-dependent manner from 3.125 to 12.5 μg/mL, where 12.5 μg/mL significantly suppressed 36.13 ± 2.1% of lipid accumulation. We postulate that inhibition of cholesterol uptake, adipogenesis, preadipocyte and adipocyte number may be utilized as treatment modalities to reduce the prevalence of lipidemia. To conclude, XNT could be a potential hypolipidemic agent to improve cardiovascular health in the future.

  11. [Effects of hydroxyl radicals on purified angiotensin I converting enzyme].

    PubMed

    Michel, B; Nirina, L B; Grima, M; Ingert, C; Coquard, C; Barthelmebs, M; Imbs, J L

    1998-08-01

    Somatic angiotensin-converting enzyme (ACE) is a protein which contains two similar domains (N and C), each possessing a functional active site. The relationship between ACE, its natural substrates and oxygen free radicals is starting to be explored. On one hand, superoxide anions production is induced by angiotensin II and on the other hand, activated polynuclear neutrophils, through free radicals generation, alter endothelial ACE activity. In this study, we examined the impact of hydroxyl radicals (.OH) on purified ACE. .OH were produced using a generator: 2,2'-azo-bis 2-amidinopropane (GRH) provided by Lara-Spiral (Fr). GRH (3 mM), in a time-dependent fashion, inhibited ACE activity. When ACE was co-incubated for 4 h with GRH, its activity decreased by 70%. Addition of dimethylthiourea (DMTU: 0.03 to 1 mM) or mannitol + methionine (20/10 mM), two sets of .OH scavengers, produced a dose-dependent protection on ACE activity. To examine whether oxidation of thiol groups in the ACE molecule could be involved in the action of GRH, the effects of thiol reducing agents: mercaptoethanol and dithiotreitol (DTT) were investigated. These compounds produced a dose-dependent and significant protection; with 100% protection at 0.2 and 0.3 mM for mercaptoethanol and at 0.1 mM for DTT. The hydrolysis of two natural and domain-specific substrates were also explored. The hydrolysis of angiotensin I preferentially cleaved by the C domain was significantly (p < 0.01) inhibited by 57, 58 and 69% in contact with 0.3, 1 and 3 mM GRH [in nmol angio II formed/min/nmol of ACE, n = 4; 35.9 +/- 0.6 (control), 15.5 +/- 2.8 (GRH : 0.3 mM), 15.1 +/- 0.5 (1), 10.9 +/- 0.6 (3)]. The hydrolysis of the hemoregulatory peptide (hp), preferential substrate for the N domain was not affected by GRH at 0.3 mM and inhibited by 28% (not significant) by 1 mM GRH [in nmol ph hydrolized/min/nmol ACE, n = 4; 12.6 +/- 1.9 (control), 14.9 (GRH : 0.3 mM), 8.3 +/- 4.0 (1). These results demonstrated that .OH

  12. 42 CFR 84.254 - Powered air-purifying respirators; requirements and tests.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... air-purifying respirators prescribed in subpart L of this part are applicable to vinyl chloride... use with powered air-purifying respirators for entry into and escape from vinyl chloride atmospheres... of 25 ppm vinyl chloride monomer at a total flow rate of 115 liters per minute for...

  13. Kinetic analysis of competition between aerosol particle removal and generation by ionization air purifiers.

    PubMed

    Alshawa, Ahmad; Russell, Ashley R; Nizkorodov, Sergey A

    2007-04-01

    Ionization air purifiers are increasingly used to remove aerosol particles from indoor air. However, certain ionization air purifiers also emit ozone. Reactions between the emitted ozone and unsaturated volatile organic compounds (VOC) commonly found in indoor air produce additional respirable aerosol particles in the ultrafine (<0.1 microm) and fine (<2.5 microm) size domains. A simple kinetic model is used to analyze the competition between the removal and generation of particulate matter by ionization air purifiers under conditions of a typical residential building. This model predicts that certain widely used ionization air purifiers may actually increase the mass concentration of fine and ultrafine particulates in the presence of common unsaturated VOC, such as limonene contained in many household cleaning products. This prediction is supported by an explicit observation of ultrafine particle nucleation events caused by the addition of D-limonene to a ventilated office room equipped with a common ionization air purifier.

  14. Effects of purified alginate sponge on the regeneration of chondrocytes: in vitro and in vivo.

    PubMed

    Song, Jeong Eun; Kim, A Ram; Lee, Cheon Jung; Tripathy, Nirmalya; Yoon, Kun Ho; Lee, Dongwon; Khang, Gilson

    2015-01-01

    Regeneration science has been studied using tissue engineering techniques due to the self-renewal difficulties of damaged or degenerated cartilage. A scaffold with biodegradability and biocompatibility features plays a key role in developing cartilage tissue similar to human biological materials. Herein, we have fabricated three-dimensional sponge using purified alginate for the regeneration of chondrocytes cells and formation of cartilage. We demonstrated that the alginate purification can effectively minimize inflammatory reaction through reducing the content of mannuronic acid causing immune rejection. Cartilage regeneration research was performed using three-dimensional non-purified and purified alginate sponges synthesized by modified Korbutt method. In vitro cell viability and specific gene expression in the cartilage cells were investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and reverse transcriptase-polymerase chain reaction (RT-PCR) after seeding chondrocytes on the as-fabricated sponges. Specific extracellular matrix (ECM) of chondrocytes, sGAG, and the content of collagen were also measured. Histological staining was carried out after purified alginate sponge seeded with chondrocytes and was implanted in subcutaneous nude mouse followed by extraction. Compared to the non-purified ones, the purified alginate sponges showed positive effects on maintaining affinities and phenotype of chondrocytes. From these results, it can be suggested that the purified alginate sponges provide a promising platform for cartilage regeneration.

  15. Pore structure of raw and purified HiPco single-walled carbon nanotubes

    NASA Astrophysics Data System (ADS)

    Cinke, Martin; Li, Jing; Chen, Bin; Cassell, Alan; Delzeit, Lance; Han, Jie; Meyyappan, M.

    2002-10-01

    Very high purity single-walled carbon nanotubes (SWNTs) were obtained from HiPco SWNT samples containing Fe particles by a two-step purification process. The raw and purified samples were characterized using high resolution transmission electron microscopy (HRTEM), Raman spectroscopy and thermogravimetric analysis (TGA). The purified sample consists of ˜0.4% Fe and the process does not seem to introduce any additional defects. The N 2 adsorption isotherm studies at 77 K reveal that the total surface area of the purified sample increases to 1587 m 2/g from 567 m 2/g for the raw material, which is the highest value reported for SWNTs.

  16. Corrugated waveguide mode purifier for TEM output in a dual-mode operation overmoded coaxial millimeter-wave generator

    NASA Astrophysics Data System (ADS)

    Bai, Zhen; Zhang, Jun; Zhong, Huihuang; Zhang, Dian

    2017-01-01

    A coaxial corrugated waveguide mode purifier is designed for a dual-mode operation overmoded coaxial millimeter-wave generator. With the purifier, the mixed TEM and TM01 modes output are purified into a pure TEM mode. Particle-in-cell (PIC) simulation shows that the purifier would not decrease the total output power of the generator, and plays an independent role to the upstream structure. Effects of mode composition ratio and phase difference on the purification ability of the purifier are also researched by both electromagnetism and PIC simulations, which show that the purifier has a certain tolerance for both the mode composition ratio and phase difference.

  17. Effects of rheological change by addition of carboxymethylcellulose in culture media of an air-lift fermentor on poly-D-3-hydroxybutyric acid productivity in autotrophic culture of hydrogen-oxidizing bacterium, Alcaligenes eutrophus.

    PubMed

    Taga, N; Tanaka, K; Ishizaki, A

    1997-03-05

    The effects of rheological change by addition of sodium carboxymethylcellulose (CMC) to culture medium in an air-lift-type fermentor on autotrophic production of poly-(D-3-hydroxybutyric acid) [P(3HB)] by two-stage culture of Alcaligenes eutrophus is investigated. Addition of 0.05% CMC increased P(3HB) production rate during the P(3HB) accumulation phase to twice that of the control culture. It was thought that addition of a small amount of CMC was beneficial for production of P(3HB) employing the air-lift fermentor under safe autotrophic culture conditions in wich oxygen concentration was maintained below 6.9% (v/v). the volumetric mass transfer coefficient (K(L)a) observed in the presence of CMC is shown to correlated with the P(3HB) production rate obtained. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 53: 529-533, 1997.

  18. Composition and potency characterization of Mycobacterium avium subsp. paratuberculosis purified protein derivatives

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium avium subsp. paratuberculosis (MAP) purified protein derivatives (PPDs) are immunologic reagents prepared from cultured filtrates of the type strain ATCC 19698. Traditional production consists of floating culture incubation at 37oC, organism inactivation by autoclaving, coarse filtrat...

  19. Novel tubular and crystalline structures in purified preparations of Newcastle disease virus. Brief report.

    PubMed

    Gowans, E J; McNulty, M S

    1979-01-01

    Hitherto undescribed tubular and crystalline structures were detected by negative contrast electron microscopy in purified preparations of Newcastle disease virus. It is suggested that these are viral in origin and are composed of aggregates of viral glycoprotein.

  20. Reproductive and neurobehavioral outcome of drinking purified water under magnesium deficiency in the rat's diet.

    PubMed

    Zeng, Hui; Shu, Wei-qun; Zhao, Qing; Chen, Qiang

    2008-05-01

    Taking magnesium deficient diet and drinking soft water (including purified water, essentially mineral free) are common consumed in the world. The present study was conducted to assess the potential combined influence of maternal drinking purified water and taking magnesium deficient diet on postnatal development and behavior in the offspring of exposed rats. Sprague-Dawley (SD) rat were assigned to four groups: group 1 fed with control diet (Mg(2+) 0.4 g/kg) and control water (Mg(2+) 12.7 mg/L), group 2 fed with control diet and purified water (Mg(2+) 0.015 mg/L), group 3 fed with magnesium deficient diet (Mg(2+) 0.2 g/kg) and control water, group 4 fed with magnesium deficient diet and purified water from 5 weeks of age of the F0 generation to 3 weeks of the F1 generation, respectively. Reproductive and neurobehavioral parameters were measured. Maternal body weights significantly decreased during treatment (before mating) and lactation periods in the group fed with magnesium deficient diet and purified water. There were no significant differences of the reproductive outcome in the groups. Offspring's body weight, development of reflexes significantly reduced in the group 4. Although it was no significant difference in the four groups, the data showed a trend toward a decreased risk for offspring body weight, neurobehavioral development as follows: group 1>group 2>group 3>group 4. Therefore, purified water cannot obviously affect the reproductive outcome when magnesium is sufficient or half of the estimated average requirement (EAR) in the diet. However, drinking purified water can decrease maternal magnesium level slightly, induce offspring's development retardation, especially when the magnesium deficiency in the diet. Furthermore, magnesium deficiency in the diet (half of the EAR) can produce growth delay and reflex development retardation in F1-offspring. Therefore, drinking purified water should be carefully considered, especially for susceptible population.

  1. Purification of uranium alloys by differential solubility of oxides and production of purified fuel precursors

    DOEpatents

    McLean, W. II; Miller, P.E.

    1997-12-16

    A method is described for purifying metallic alloys of uranium for use as nuclear reactor fuels in which the metal alloy is first converted to an oxide and then dissolved in nitric acid. Initial removal of metal oxide impurities not soluble in nitric acid is accomplished by filtration or other physical means. Further purification can be accomplished by carbonate leaching of uranyl ions from the partially purified solution or using traditional methods such as solvent extraction. 3 figs.

  2. Purification of uranium alloys by differential solubility of oxides and production of purified fuel precursors

    DOEpatents

    McLean, II, William; Miller, Philip E.

    1997-01-01

    A method for purifying metallic alloys of uranium for use as nuclear reactor fuels in which the metal alloy is first converted to an oxide and then dissolved in nitric acid. Initial removal of metal oxide impurities not soluble in nitric acid is accomplished by filtration or other physical means. Further purification can be accomplished by carbonate leaching of uranyl ions from the partially purified solution or using traditional methods such as solvent extraction.

  3. An Experiment with Air Purifiers in Delhi during Winter 2015-2016.

    PubMed

    Vyas, Sangita; Srivastav, Nikhil; Spears, Dean

    2016-01-01

    Particulate pollution has important consequences for human health, and is an issue of global concern. Outdoor air pollution has become a cause for alarm in India in particular because recent data suggest that ambient pollution levels in Indian cities are some of the highest in the world. We study the number of particles between 0.5μm and 2.5μm indoors while using affordable air purifiers in the highly polluted city of Delhi. Though substantial reductions in indoor number concentrations are observed during air purifier use, indoor air quality while using an air purifier is frequently worse than in cities with moderate pollution, and often worse than levels observed even in polluted cities. When outdoor pollution levels are higher, on average, indoor pollution levels while using an air purifier are also higher. Moreover, the ratio of indoor air quality during air purifier use to two comparison measures of air quality without an air purifier are also positively correlated with outdoor pollution levels, suggesting that as ambient air quality worsens there are diminishing returns to improvements in indoor air quality during air purifier use. The findings of this study indicate that although the most affordable air purifiers currently available are associated with significant improvements in the indoor environment, they are not a replacement for public action in regions like Delhi. Although private solutions may serve as a stopgap, reducing ambient air pollution must be a public health and policy priority in any region where air pollution is as high as Delhi's during the winter.

  4. An Experiment with Air Purifiers in Delhi during Winter 2015-2016

    PubMed Central

    Vyas, Sangita

    2016-01-01

    Particulate pollution has important consequences for human health, and is an issue of global concern. Outdoor air pollution has become a cause for alarm in India in particular because recent data suggest that ambient pollution levels in Indian cities are some of the highest in the world. We study the number of particles between 0.5μm and 2.5μm indoors while using affordable air purifiers in the highly polluted city of Delhi. Though substantial reductions in indoor number concentrations are observed during air purifier use, indoor air quality while using an air purifier is frequently worse than in cities with moderate pollution, and often worse than levels observed even in polluted cities. When outdoor pollution levels are higher, on average, indoor pollution levels while using an air purifier are also higher. Moreover, the ratio of indoor air quality during air purifier use to two comparison measures of air quality without an air purifier are also positively correlated with outdoor pollution levels, suggesting that as ambient air quality worsens there are diminishing returns to improvements in indoor air quality during air purifier use. The findings of this study indicate that although the most affordable air purifiers currently available are associated with significant improvements in the indoor environment, they are not a replacement for public action in regions like Delhi. Although private solutions may serve as a stopgap, reducing ambient air pollution must be a public health and policy priority in any region where air pollution is as high as Delhi’s during the winter. PMID:27978542

  5. Investigating the characteristic strength of flocs formed from crude and purified Hibiscus extracts in water treatment.

    PubMed

    Jones, Alfred Ndahi; Bridgeman, John

    2016-10-15

    The growth, breakage and re-growth of flocs formed using crude and purified seed extracts of Okra (OK), Sabdariffa (SB) and Kenaf (KE) as coagulants and coagulant aids was assessed. The results showed floc size increased from 300 μm when aluminium sulphate (AS) was used as a coagulant to between 696 μm and 722 μm with the addition of 50 mg/l of OK, KE and SB crude samples as coagulant aids. Similarly, an increase in floc size was observed when each of the purified proteins was used as coagulant aid at doses of between 0.123 and 0.74 mg/l. The largest floc sizes of 741 μm, 460 μm and 571 μm were obtained with a 0.123 mg/l dose of purified Okra protein (POP), purified Sabdariffa (PSP) and purified Kenaf (PKP) respectively. Further coagulant aid addition from 0.123 to 0.74 mg/l resulted in a decrease in floc size and strength in POP and PSP. However, an increase in floc strength and reduced d50 size was observed in PKP at a dose of 0.74 mg/l. Flocs produced when using purified and crude extract samples as coagulant aids exhibited high recovery factors and strength. However, flocs exhibited greater recovery post-breakage when the extracts were used as a primary coagulant. It was observed that the combination of purified proteins and AS improved floc size, strength and recovery factors. Therefore, the applications of Hibiscus seeds in either crude or purified form increases floc growth, strength, recoverability and can also reduce the cost associated with the import of AS in developing countries.

  6. [Diurnal variations in purifying-tanks when use Pontederia cordata treating the Malodorous River water].

    PubMed

    Chen, Jian-jun; Lu, Xiao-ming; Lu, Shao-yong; Jin, Xiang-can; Huang, Min-sheng; Zhang, Yong; Zhao, Feng

    2009-12-01

    Aquatic plants (Ponsederie cordaza) were waked in two purifying-tanks to investigate the effects of illumination intensity and aeration on diurnal variations of Chla, SP, POD of Ponsederia cordaza and pH, DO, COD, NH4+ -N, TP of water from purifying-tanks when treating the malodorous river water at seven different times, another blank purifying-tank was set as a control. Comparative studies and correlation analysis of these different indicators were carried out to improve the plants working efficiency and provide scientific basis for optimal operation of plant purifying-tanks. Results showed that all indicators affected by changes of light, TP shows best correlation coefficient Cr = 0.93, p < 0.01) of physicochemical indicators and SP behaves best correlation coefficient Cr = 0.91 , p < 0.01) of plant physiology indicators in non-aeration purifying-tank;aeration is necessary as diurnal average of DO shows an increase of 0.13 mg/L by treatment of plant meanwhile 1.8 mg/L by plant with aeration,purifying-tanks with aeration got 7.1%, 6.3% higher removing rates of COD, NH4+ -N and 38% less TP removing rate than non-aeration plant purifying-tanks (p < 0.01); with aeration treatment, significant reduction of Chla, SP content (p < 0.05) and increase of POD activity (p < 0.05) observed in plants; the changes of illumination intensity and aeration can significantly affect physiological characteristics of plants and should be considered carefully and need further study when treating malodorous river water by plant purifying-tanks.

  7. Effect of Purified Mushroom Tyrosinase on Melanin Content and Melanogenic Protein Expression

    PubMed Central

    Ali, Ayesha S.

    2016-01-01

    In mammalian melanocytes, melanosome is a highly specialized organelle where melanin is synthesized. Melanin synthesis is controlled by tyrosinase, the vital enzyme in melanogenic pathway. The present investigation is based on an effect of purified mushroom tyrosinase of Agaricus bisporus on B16F10 melanocytes for the melanin production via blocking pigment cell machinery. Using B16F10 melanocytes showed that the stimulation of melanogenesis by purified tyrosinase is due to increased tyrosinase absorption. Cellular tyrosinase activity and melanin content in B16F10 melanocytes were increased by purified tyrosinase in a dose-dependent manner. Western blot analysis revealed that cellular tyrosinase levels were enhanced after treatment with purified tyrosinase for 48 hours. Furthermore, tyrosinase induced phosphorylation of cyclic adenosine monophosphate (cAMP) response element-binding protein (CREB) in a dose-dependent manner. The purified tyrosinase-mediated increase of tyrosinase activity was significantly attenuated by H89, LY294002, Ro-32-0432, and PD98059, cAMP-dependent protein kinase inhibitors. The results indicate that purified tyrosinase can be used as contestant for the treatment of vitiligous skin conditions. PMID:27699070

  8. Quantitation of tyrosine hydroxylase, protein levels: Spot immunolabeling with an affinity-purified antibody

    SciTech Connect

    Haycock, J.W. )

    1989-09-01

    Tyrosine hydroxylase was purified from bovine adrenal chromaffin cells and rat pheochromocytoma using a rapid (less than 2 days) procedure performed at room temperature. Rabbits were immunized with purified enzyme that was denatured with sodium dodecylsulfate, and antibodies to tyrosine hydroxylase were affinity-purified from immune sera. A Western blot procedure using the affinity-purified antibodies and {sup 125}I-protein A demonstrated a selective labeling of a single Mr approximately 62,000 band in samples from a number of different tissues. The relative lack of background {sup 125}I-protein A binding permitted the development of a quantitative spot immunolabeling procedure for tyrosine hydroxylase protein. The sensitivity of the assay is 1-2 ng of enzyme. Essentially identical standard curves were obtained with tyrosine hydroxylase purified from rat pheochromocytoma, rat corpus striatum, and bovine adrenal medulla. An extract of PC 12 cells (clonal rat pheochromocytoma cells) was calibrated against purified rat pheochromocytoma tyrosine hydroxylase and used as an external standard against which levels of tyrosine hydroxylase in PC12 cells and other tissue were quantified. With this procedure, qualitative assessment of tyrosine hydroxylase protein levels can be obtained in a few hours and quantitative assessment can be obtained in less than a day.

  9. Caspase Inhibitors of the P35 Family Are More Active When Purified from Yeast than Bacteria

    PubMed Central

    Brand, Ingo L.; Civciristov, Srgjan; Taylor, Nicole L.; Talbo, Gert H.; Pantaki-Eimany, Delara; Levina, Vita; Clem, Rollie J.; Perugini, Matthew A.; Kvansakul, Marc; Hawkins, Christine J.

    2012-01-01

    Many insect viruses express caspase inhibitors of the P35 superfamily, which prevent defensive host apoptosis to enable viral propagation. The prototypical P35 family member, AcP35 from Autographa californica M nucleopolyhedrovirus, has been extensively studied. Bacterially purified AcP35 has been previously shown to inhibit caspases from insect, mammalian and nematode species. This inhibition occurs via a pseudosubstrate mechanism involving caspase-mediated cleavage of a “reactive site loop” within the P35 protein, which ultimately leaves cleaved P35 covalently bound to the caspase's active site. We observed that AcP35 purifed from Saccharomyces cerevisae inhibited caspase activity more efficiently than AcP35 purified from Escherichia coli. This differential potency was more dramatic for another P35 family member, MaviP35, which inhibited human caspase 3 almost 300-fold more potently when purified from yeast than bacteria. Biophysical assays revealed that MaviP35 proteins produced in bacteria and yeast had similar primary and secondary structures. However, bacterially produced MaviP35 possessed greater thermal stability and propensity to form higher order oligomers than its counterpart purified from yeast. Caspase 3 could process yeast-purified MaviP35, but failed to detectably cleave bacterially purified MaviP35. These data suggest that bacterially produced P35 proteins adopt subtly different conformations from their yeast-expressed counterparts, which hinder caspase access to the reactive site loop to reduce the potency of caspase inhibition, and promote aggregation. These data highlight the differential caspase inhibition by recombinant P35 proteins purified from different sources, and caution that analyses of bacterially produced P35 family members (and perhaps other types of proteins) may underestimate their activity. PMID:22720082

  10. [An easy way to purify the inclusion body protein with high purity from prokaryotic expression cells].

    PubMed

    Liu, Rong; Zhong, Qin-Ping; Jiang, Ming-Sen; Dong, Hui-Fen

    2011-10-01

    To clone partial ORF of SjBMP and to construct the recombinant SjBMP-pET-28a(+) plasmids, and then to transform them into the competent cells E. coli BL21 (DE3), finally a positive clone was used to be induced by IPTG. The bacterial aggregates with target protein expressed as inclusion bodies were purified by the methods of Ni(2+)-NTA affinity purification under denaturation condition and SDS-PAGE gel extraction. The purified protein was used to immune rabbits and make antiserum against the SjBMP, and the antiserum were then used to identify the rSjBMP by Western blotting. The target protein obtained by Ni(2+)-NTA Agarose affinity purification was not pure with unspecific proteins, but the protein further purified by SDS-PAGE gel extraction and the dialysis bag horizontal electrophoresis was quite pure, and the recovery rate was more than 11.0%. Meanwhile, Western blotting was used to identify the recombinant SjBMP protein by antiserum, only a specific single strip appeared, which suggested the protein purified by this method kept its antigenicity, and could be used for common immunological studies. Therefore, the SDS-PAGE gel extraction combining with electroosmosis and dialysis recycling are good and easy to purify the inclusion body proteins.

  11. Quantification of ozone levels in indoor environments generated by ionization and ozonolysis air purifiers.

    PubMed

    Britigan, Nicole; Alshawa, Ahmad; Nizkorodov, Sergey A

    2006-05-01

    Indoor air purifiers are advertised as safe household products for health-conscious individuals, especially for those suffering from allergies and asthma. However, certain air purifiers produce ozone (O3) during operation, either intentionally or as a byproduct of air ionization. This is a serious concern, because O3 is a criteria air pollutant regulated by health-related federal and state standards. Several types of air purifiers were tested for their ability to produce ozone in various indoor environments at 40-50% relative humidity, including office rooms, bathrooms, bedrooms, and cars. O3 levels generated by personal wearable air purifiers were also tested. In many cases, O3 concentrations were well in excess of public and/or industrial safety levels established by U.S. Environmental Protection Agency, California Air Resources Board, and Occupational Safety and Health Administration. Simple kinetic equations were obtained that can predict the steady-state level of O3 in a room from the O3 emission rate of the air purifier and the first-order decay rate of O3 in the room. The additivity of O3 levels generated by independent O3 generators was experimentally demonstrated.

  12. Somatic Mutation Patterns in Hemizygous Genomic Regions Unveil Purifying Selection during Tumor Evolution

    PubMed Central

    Basu, Swaraj; Larsson, Erik

    2016-01-01

    Identification of cancer driver genes using somatic mutation patterns indicative of positive selection has become a major goal in cancer genomics. However, cancer cells additionally depend on a large number of genes involved in basic cellular processes. While such genes should in theory be subject to strong purifying (negative) selection against damaging somatic mutations, these patterns have been elusive and purifying selection remains inadequately explored in cancer. Here, we hypothesized that purifying selection should be evident in hemizygous genomic regions, where damaging mutations cannot be compensated for by healthy alleles. Using a 7,781-sample pan-cancer dataset, we first confirmed this in POLR2A, an essential gene where hemizygous deletions are known to confer elevated sensitivity to pharmacological suppression. We next used this principle to identify several genes and pathways that show patterns indicative of purifying selection to avoid deleterious mutations. These include the POLR2A interacting protein INTS10 as well as genes involved in mRNA splicing, nonsense-mediated mRNA decay and other RNA processing pathways. Many of these genes belong to large protein complexes, and strong overlaps were observed with recent functional screens for gene essentiality in human cells. Our analysis supports that purifying selection acts to preserve the remaining function of many hemizygously deleted essential genes in tumors, indicating vulnerabilities that might be exploited by future therapeutic strategies. PMID:28027311

  13. The control of biofilm formation by hydrodynamics of purified water in industrial distribution system.

    PubMed

    Florjanič, Maja; Kristl, Julijana

    2011-02-28

    Systems for storage and distribution of purified water at ambient temperature are highly susceptible to microbial contamination. The water flow, microbial content and chemical quality of the purified water in an industrial water system have been simulated in a biofilm annular reactor (BAR) to study the impact of different hydrodynamic conditions on biofilm development. Our results reveal the potential of stagnant purified water at total organic compounds (TOC) below 50ppb to develop biofilm that allows detachment of planktonic bacteria and colonization of new surfaces within 24h. However, under constant water flow over 7 days, the growth of initial biofilm was 40 times less, fewer bacteria were detached, and new surfaces were colonized to a lesser extent. Heterotrophic plate counts (HPCs) in biofilm were highly positively correlated with numbers of detached planktonic bacteria in effluent water. The study shows that the hydrodynamic conditions and level of planktonic HPC in water are critical for the development of biofilm at very low TOC. The results in the BAR agreed well with those from regular industrial microbial monitoring of purified water. To conclude, the BAR successfully simulates biofilm growth and can be used to establish an effective biofilm control strategy. However, the microbial quality of purified water in industrial system is a constant challenge; any increase of HPC in effluent water is a sign to take steps against excessive microbial growth.

  14. [Chromatographic and spectroscopic characterization of phycocyanin and its subunits purified from Anabaena variabilis CCC421].

    PubMed

    Chakdar, N; Sakha, S; Pabbi, S

    2014-01-01

    Phycocyanin, a high value pigment was purified from diazotrophic cyanobacteria Anabaena variabilis CCC421 using a strategy involving ammonium sulfate precipitation, dialysis and anion exchange chromatography using DEAE-cellulose column. 36% phycocyanin with a purity of 2.75 was recovered finally after anion exchange chromatography. Purified phycocyanin was found to contain 2 subunits of 17 and 18 kDa which were identified as a-and (3 subunits by SDS-PAGE and MALDI-TOE HPLC method using a C5 column coupled with fluorescence or photodiode-based detection was also developed to separate and detect the A. variabilis CCC421 phycocyanin subunits. The fluorescence method was more sensitive than photodiode one. The purified phycocyanin from A. variabilis CCC421 as well as its subunits was characterized with respect to absorption and IR spectra. Spectral characterization of the subunits revealed that alpha and beta subunits contained one and two phycocyanobilin groups as chromophores, respectively.

  15. Purified plasminogen activating factor produced by malignant lymphoid cells abrogates lymphocyte cytotoxicity.

    PubMed Central

    Sundar, S K; Bergeron, J; Menezes, J

    1984-01-01

    Immunosuppression is a generally observed phenomenon in patients with malignancies. Here we report that plasminogen activating factor (PAF) produced by human (P3HR-1) and simian (B95-8) lymphoid cells of malignant origin abrogates lymphocyte cytotoxicity. PAF has been purified from Epstein-Barr (EB) virus genome carrying lymphocyte cytotoxicity. PAF has been purified from Epstein-Barr (EB) virus genome carrying lymphoid lines by affinity chromatography using lysine-Sepharose columns. Purified PAF consistently inhibited Killer cell activity against the following targets: K-562, EB virus superinfected Raji cells and in vitro EB virus transformed autologous B lymphocytes. Furthermore PAF also inhibited the antibody-dependent cellular cytotoxicity. The results presented also indicate that PAF affects the effector lymphocytes and not the target cells. Taken together, these observations emphasize the importance of factors such as PAF, released by malignant cells, as inhibitors/modulators of immune mechanisms effective against tumour cells. PMID:6430612

  16. Electrorefiner system for recovering purified metal from impure nuclear feed material

    DOEpatents

    Berger, John F.; Williamson, Mark A.; Wiedmeyer, Stanley G.; Willit, James L.; Barnes, Laurel A.; Blaskovitz, Robert J.

    2015-10-06

    An electrorefiner system according to a non-limiting embodiment of the present invention may include a vessel configured to maintain a molten salt electrolyte and configured to receive a plurality of alternately arranged cathode and anode assemblies. The anode assemblies are configured to hold an impure nuclear feed material. Upon application of the power system, the impure nuclear feed material is anodically dissolved and a purified metal is deposited on the cathode rods of the cathode assemblies. A scraper is configured to dislodge the purified metal deposited on the cathode rods. A conveyor system is disposed at a bottom of the vessel and configured to remove the dislodged purified metal from the vessel.

  17. Effect of home-used water purifier on fluoride concentration of drinking water in southern Iran

    PubMed Central

    Jaafari-Ashkavandi, Zohreh; Kheirmand, Mehdi

    2013-01-01

    Background: Fluoride in drinking water plays a key role in dental health. Due to the increasing use of water-purifier, the effect of these devices on fluoride concentration of drinking water was evaluated. Materials and Methods: Drinking water samples were collected before and after passing through a home water-purifier, from four different water sources. The fluoride, calcium and magnesium concentration of the samples were measured using the quantitative spectrophotometery technique. Data were analyzed by the Wilcoxon test. P value < 0.1 was considered as significant. Results: The result showed that the concentration of fluoride was 0.05-0.61 ppm before purification and was removed completely afterward. Furthermore, other ions reduced significantly after treatment by the water purifier. Conclusion: This study revealed that this device decreases the fluoride content of water, an issue which should be considered in low and high-fluoridated water sources. PMID:24130584

  18. Purifying Selection, Drift, and Reversible Mutation with Arbitrarily High Mutation Rates

    PubMed Central

    Charlesworth, Brian; Jain, Kavita

    2014-01-01

    Some species exhibit very high levels of DNA sequence variability; there is also evidence for the existence of heritable epigenetic variants that experience state changes at a much higher rate than sequence variants. In both cases, the resulting high diversity levels within a population (hyperdiversity) mean that standard population genetics methods are not trustworthy. We analyze a population genetics model that incorporates purifying selection, reversible mutations, and genetic drift, assuming a stationary population size. We derive analytical results for both population parameters and sample statistics and discuss their implications for studies of natural genetic and epigenetic variation. In particular, we find that (1) many more intermediate-frequency variants are expected than under standard models, even with moderately strong purifying selection, and (2) rates of evolution under purifying selection may be close to, or even exceed, neutral rates. These findings are related to empirical studies of sequence and epigenetic variation. PMID:25230951

  19. Immunogenicity of a purified fragment of 17D yellow fever envelope protein.

    PubMed

    Brandriss, M W; Schlesinger, J J; Walsh, E E

    1990-06-01

    Information on the immunogenic properties of purified flavivirus proteins may be useful in the development of recombinant or synthetic peptide vaccines. Using a monoclonal antibody, an attempt was made to purify the envelope (E) protein of 17D yellow fever virus (17D YF) by affinity chromatography. The purified material could not be identified as intact E protein but it did bear antigenic determinants of E as determined by selective reactivity with anti-E monoclonal antibodies. Rabbits immunized with this material produced antibodies that neutralized 17D YF and dengue-2 viruses in comparable titers, indicating that cross-reactive antigenic determinants were preserved. Immunization of mice resulted in protection against intracerebral challenge with 17D YF.

  20. Enhanced purity, activity and structural integrity of yeast ribosomes purified using a general chromatographic method.

    PubMed

    Leshin, Jonathan A; Rakauskaitė, Rasa; Dinman, Jonathan D; Meskauskas, Arturas

    2010-01-01

    One of the major challenges facing researchers working with eukaryotic ribosomes lies in their lability relative to their eubacterial and archael counterparts. In particular, lysis of cells and purification of eukaryotic ribosomes by conventional differential ultracentrifugation methods exposes them for long periods of time to a wide range of co-purifying proteases and nucleases, negatively impacting their structural integrity and functionality. A chromatographic method using a cysteine charged Sulfolink resin was adapted to address these problems. This fast and simple method significantly reduces co-purifying proteolytic and nucleolytic activities, producing good yields of highly biochemically active yeast ribosomes with fewer nicks in their rRNAs. In particular, the chromatographic purification protocol significantly improved the quality of ribosomes isolated from mutant cells. This method is likely applicable to mammalian ribosomes as well. The simplicity of the method, and the enhanced purity and activity of chromatographically purified ribosome represents a significant technical advancement for the study of eukaryotic ribosomes.

  1. Guanine nucleotide regulatory protein co-purifies with the D/sub 2/-dopamine receptor

    SciTech Connect

    Senogles, S.E.; Caron, M.G.

    1986-05-01

    The D/sub 2/-dopamine receptor from bovine anterior pituitary was purified approx.1000 fold by affinity chromatography on CMOS-Sepharose. Reconstitution of the affinity-purified receptor into phospholipid vesicles revealed the presence of high and low affinity agonist sites as detected by N-n-propylnorapomorphine (NPA) competition experiments with /sup 3/H-spiperone. High affinity agonist binding could be converted to the low affinity form by guanine nucleotides, indicating the presence of an endogenous guanine nucleotide binding protein (N protein) in the affinity-purified D/sub 2/ receptor preparations. Furthermore, this preparation contained an agonist-sensitive GTPase activity which was stimulated 2-3 fold over basal by 10 ..mu..M NPA. /sup 35/S-GTP..gamma..S binding to these preparations revealed a stoichiometry of 0.4-0.7 mole N protein/mole receptor, suggesting the N protein may be specifically coupled with the purified D/sub 2/-dopamine receptor and not present as a contaminant. Pertussis toxin treatment of the affinity purified receptor preparations prevented high affinity agonist binding, as well as agonist stimulation of the GTPase activity, presumably by inactivating the associated N protein. Pertussis toxin lead to the ADP-ribosylation of a protein of 39-40K on SDS-PAGE. These findings indicate that an endogenous N protein, N/sub i/ or N/sub o/, co-purifies with the D/sub 2/-dopamine receptor which may reflect a precoupling of this receptor with an N protein within the membranes.

  2. Presence of glycosaminoglycans in purified AA type amyloid fibrils associated with juvenile rheumatoid arthritis.

    PubMed Central

    Magnus, J H; Husby, G; Kolset, S O

    1989-01-01

    Previous studies have strongly suggested an association between glycosaminoglycans and tissue deposits of amyloid. The present study was aimed at studying this association in purified preparations of hepatic amyloid fibrils obtained from human AA type secondary amyloidosis. Glycosaminoglycans were isolated by gradient ion exchange chromatography of purified amyloid fibrils treated with pronase. Degradation with specific enzymes identified the glycosaminoglycans as chondroitin sulphate, dermatan sulphate, and heparin/heparan sulphate. The total amount of glycosaminoglycans specifically coisolated with the amyloid fibrils was 15 micrograms/mg fibril weight. The presence of glycosaminoglycans in amyloid may play a part in the incorporation of structurally diverse protein precursors into amyloid fibrils of identical ultrastructure. PMID:2930277

  3. Hematemesis With Gastric Laceration After Tattooing a Polyp With Purified Carbon: A Review of the Literature

    PubMed Central

    Gonzalez-Tallon, Ana Isabel; Rivero-Fernandez, Miguel; Calvo-Ramos, Irina; Diaz-Sanchez, Antonio; del Rosario Gonzalez-Alonso, Maria; la Fuente-Briongos, Elsa De; Manzano-Fernandez, Rebeba; Moya-Valverde, Eloisa; Riesco-Lopez, Jose Maria; Campos-Cantero, Rocio

    2017-01-01

    Endoscopic tattooing is a simple and effective technique for marking small lesions, so they can be localized during surgery or in later endoscopies. Various agents can be used such as India ink or a solution of purified carbon particles. The number of complications from tattooing is relatively small, but not rare. The majority of the literature on the subject refers to complications in the colon. We present a case of gastric bleeding secondary to a laceration following tattooing with purified carbon, and a literature review. PMID:28270877

  4. Using Air-Purifying Respirators. Module 9. Vocational Education Training in Environmental Health Sciences.

    ERIC Educational Resources Information Center

    Consumer Dynamics Inc., Rockville, MD.

    This module, one of 25 on vocational education training for careers in environmental health occupations, contains self-instructional materials on using air-purifying respirators. Following guidelines for students and instructors and an introduction that explains what the student will learn are three lessons: (1) describing how air flows through an…

  5. [Serodiagnosis of schistosomiasis mansoni using an egg extract semi-purified by precipitation with ammonium sulfate].

    PubMed

    Ouattara, S A; Sauneron, M F; Tribouley-Duret, J; Tribouley, J

    1986-01-01

    Fifty one sera from bilharziosis patients and thirty control sera were examined with a Schistosoma mansoni egg antigen purified by precipitation with ammonium sulphate at 50% saturation. Sensitivity and specificity were good and showed a good correlation with results obtained by MSA1 antigen, but antigen tested is far more easier to prepare than MSA1.

  6. A resource-saving technology for purifying oil-contaminated effluent waters from thermal power stations

    NASA Astrophysics Data System (ADS)

    Borodai, E. N.; Nikolaeva, L. A.; Laptev, A. G.

    2011-07-01

    A modified resource-saving scheme for purifying effluent waters produced at thermal stations from petroleum products is described taking the Kazan TETs-3 cogeneration station as an example, in which the sludge from chemical water treatment is used as adsorbent.

  7. Kosmotropes enhance the yield of antibody purified by affinity chromatography using immobilized bacterial immunoglobulin binding proteins.

    PubMed

    Ngo, That T; Narinesingh, Dyer

    2008-01-01

    The yield of antibody purified using affinity chromatography on immobilized Protein A or Protein G was increased up to 5-fold (500%) by including kosmotropic salts in the binding buffer. The binding buffer is used to equilibrate the affinity column before applying a sample to the column and also to dilute the sample prior to loading onto the affinity column to optimize conditions for a maximal binding of antibodies to affinity gels. In this study, the kosmotropic salts that were effective in greatly increasing antibody binding to Protein A included both inorganic and organic salts of ammonium; sodium; or potassium sulfate, phosphate, polycarboxylates; for example, succinate, citrate, isocitrate, N-(2-hydroxyethylene diamine triacetate (HEDTA), ethylene diamine tetraacetate (EDTA), and ethylene glycol-O,O'-bis(2-aminoethyl)-N,N,N'N'-tetra acetate(EGTA). On an equal-molar basis, the greater the number of carboxylic groups within the polycarboxylate molecule, the greater the increase in the yield of the purified antibody that was observed. The data show that kosmotropes can be used as effective additives to enhance the binding of immunoglobulins to Protein A or Protein G gels with a resultant increase in the yield of the purified antibodies. Thus, it appears that strongly hydrated anions (citrate, sulfate, and phosphate) and weakly hydrated cations (ammonium, potassium) increase the yield of antibody purified on either Protein A or Protein G affinity gels.

  8. Air purifiers that diffuse reactive oxygen species potentially cause DNA damage in the lung.

    PubMed

    Kawamoto, Kosuke; Sato, Itaru; Yoshida, Midori; Tsuda, Shuji

    2010-12-01

    Several appliance manufacturers have recently released new type air purifiers that can disinfect bacteria, fungi and viruses by diffusing reactive oxygen species (ROS) into the air. In this study, mice were exposed to the outlet air from each of 3 air purifiers from different manufacturers (A, B, C), and the lung was examined for DNA damage, lipid peroxidation and histopathology to confirm the safety of these air purifiers. Neither abnormal behavior during exposure nor gross abnormality at necropsy was observed. No histopathological changes were also observed in the lung. However, significant increase of DNA damage was detected by the comet assay in the lung immediately after the direct exposure for 48 hr to models A and B, and for 16 hr to model B. As for model B, DNA migration was also increased by 2 hr exposure in a 1 m(3) plastic chamber but not by 48 hr exposure in a room (12.6 m(3)). Model C did not cause DNA damage. Lipid peroxidation and 8-hydroxy deoxyguanosine (8-OH-dG) was not increased under the conditions DNA damage was detected by the comet assay. The present results revealed that some models of air purifiers that diffuse ROS potentially cause DNA damage in the lung although the mechanism was left unsolved.

  9. Antifungal properties of wheat histones (H1-H4) and purified wheat histone H1

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wheat (Triticum sp.) histones H1, H2, H3, and H4 were extracted. H1 was further purified. Their activities against fungi with varying degrees of wheat pathogenicity were determined. They included Aspergillus flavus, A. fumigatus, A. niger, F. oxysporum, F. verticillioides, F. solani, F. graminearu...

  10. Immunoprotection in sheep against Haemonchus contortus using its thiol-purified excretory/secretory proteins.

    PubMed

    Arunkumar, Selvarayar

    2012-01-01

    Excretory/Secretory antigen was prepared by culturing live adult worms of Haemonchus contortus in RPMI 1640 medium at a concentration of 50 worms per mL in a culture-flask at 37 ˚C for 24 hr and the culture supernatant was used as antigen. The E/S antigen was purified by thiol-sepharose affinity chromatography. On western blot analysis, it was demonstrated that thiol-purified antigen showed a single reactive band at 66 kDa. In immunization trial, sheep were administered intramuscularly with 500 µg of thiol-purified excretory/secretory antigen along with montanide as adjuvant on day 0, 30 and 60. On ELISA, it was observed that the mean absorbance values were significantly (p ≤ 0.01) higher up to 20 weeks post immunization in Group-I (purified antigen) compared to Group- II (unimmunized control). Further, the mean EPG values was lower in Group I (200.00 ± 40.82 to 400.00 ± 91.29) than Group II (2200.00 ± 108.01 to 5100.00 ± 169.56) and the percentage reduction in mean fecal egg counts was 88.50%. Similarly, the mean abomasal worm counts was lower in Group I (808.33 ± 78.29) than Group II (3280.00 ± 147.19) and the percentage reduction in mean abomasal worm count was 75.40%.

  11. A Method for Purifying Obligate Intracellular Coxiella burnetii that Employs Digitonin Lysis of Host Cells

    PubMed Central

    Cockrell, Diane C.; Beare, Paul A.; Fischer, Elizabeth R.; Howe, Dale; Heinzen, Robert. A.

    2008-01-01

    Purification of the obligate intracellular bacterium Coxiella burnetii requires physical disruption of infected cells. Here we describe a gentle and safe digitonin lysis procedure to release C. burnetii from infected cells. The purity, yield, and infectivity of digitonin-prepped organisms are comparable to that of organisms purified using cell lysis by sonication. PMID:18242746

  12. [Experimental detection of the paraspecific effects of purified and inactivated poxviruses].

    PubMed

    Mayr, A; Büttner, M; Wolf, G; Meyer, H; Czerny, C

    1989-03-01

    The paraspecific effect of pox viruses i.e. stimulation of the nonspecific part of the complex immune system, was demonstrated in vitro and in vivo for 3 different types of virus (vaccinia virus, avipox virus, parapox virus). By means of purification, concentration and inactivation, it was possible to prove that the paraspecific effect is brought about by the pox virus particle per se. The effect is induced by certain epitopes in the complex structure of the virus and is independent of virus multiplication. Both purified pox viruses and inactivated, purified pox viruses stimulate NK cell activity, the CSA and the formation of interferon, and reach higher efficacy indices in a challenge test with baby mice using stomatitis vesicularis virus than non-purified and non-inactivated cell culture material. The degree of purity of the pox virus preparations was established using the electron microscope and monoclonal antibodies. The purified and concentrated pox virus preparations proved to be pyrogen-free; they were not toxic to baby mice in vivo. An endotoxin-induced effect could be ruled out. The broad paraspecific effect of pox viruses and their structural components lends itself to use as paramunity inducers in prophylaxis and therapy.

  13. 42 CFR 84.181 - Non-powered air-purifying particulate filter efficiency level determination.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... sodium chloride or equivalent solid aerosol at 25 ±5 °C and relative humidity of 30 ±10 percent that has... further decrease in efficiency. (g) The sodium chloride test aerosol shall have a particle size...-purifying particulate respirator model shall be tested for filter efficiency against: (1) A solid...

  14. 42 CFR 84.181 - Non-powered air-purifying particulate filter efficiency level determination.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... sodium chloride or equivalent solid aerosol at 25 ±5 °C and relative humidity of 30 ±10 percent that has... further decrease in efficiency. (g) The sodium chloride test aerosol shall have a particle size...-purifying particulate respirator model shall be tested for filter efficiency against: (1) A solid...

  15. 42 CFR 84.181 - Non-powered air-purifying particulate filter efficiency level determination.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... sodium chloride or equivalent solid aerosol at 25 ±5 °C and relative humidity of 30 ±10 percent that has... further decrease in efficiency. (g) The sodium chloride test aerosol shall have a particle size...-purifying particulate respirator model shall be tested for filter efficiency against: (1) A solid...

  16. 42 CFR 84.181 - Non-powered air-purifying particulate filter efficiency level determination.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... sodium chloride or equivalent solid aerosol at 25 ±5 °C and relative humidity of 30 ±10 percent that has... further decrease in efficiency. (g) The sodium chloride test aerosol shall have a particle size...-purifying particulate respirator model shall be tested for filter efficiency against: (1) A solid...

  17. 68. Interior view in pit "B" showing air compressor/purifier on ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    68. Interior view in pit "B" showing air compressor/purifier on left, and entry door to pit in center, with fallout shelter/escapr route on right, looking east - Nike Missile Battery MS-40, County Road No. 260, Farmington, Dakota County, MN

  18. Biological properties of purified recombinant HCV particles with an epitope-tagged envelope

    SciTech Connect

    Takahashi, Hitoshi; Akazawa, Daisuke; Kato, Takanobu; Date, Tomoko; Shirakura, Masayuki; Nakamura, Noriko; Mochizuki, Hidenori; Tanaka-Kaneko, Keiko; Sata, Tetsutaro; Tanaka, Yasuhito; Mizokami, Masashi; Suzuki, Tetsuro; Wakita, Takaji

    2010-05-14

    To establish a simple system for purification of recombinant infectious hepatitis C virus (HCV) particles, we designed a chimeric J6/JFH-1 virus with a FLAG (FL)-epitope-tagged sequence at the N-terminal region of the E2 hypervariable region-1 (HVR1) gene (J6/JFH-1/1FL). We found that introduction of an adaptive mutation at the potential N-glycosylation site (E2N151K) leads to efficient production of the chimeric virus. This finding suggests the involvement of glycosylation at Asn within the envelope protein(s) in HCV morphogenesis. To further analyze the biological properties of the purified recombinant HCV particles, we developed a strategy for large-scale production and purification of recombinant J6/JFH-1/1FL/E2N151K. Infectious particles were purified from the culture medium of J6/JFH-1/1FL/E2N151K-infected Huh-7 cells using anti-FLAG affinity chromatography in combination with ultrafiltration. Electron microscopy of the purified particles using negative staining showed spherical particle structures with a diameter of 40-60 nm and spike-like projections. Purified HCV particle-immunization induced both an anti-E2 and an anti-FLAG antibody response in immunized mice. This strategy may contribute to future detailed analysis of HCV particle structure and to HCV vaccine development.

  19. Specific activity and isotope abundances of strontium in purified strontium-82

    SciTech Connect

    Fitzsimmons, J. M.; Medvedev, D. G.; Mausner, L. F.

    2015-11-12

    A linear accelerator was used to irradiate a rubidium chloride target with protons to produce strontium-82 (Sr-82), and the Sr-82 was purified by ion exchange chromatography. The amount of strontium associated with the purified Sr-82 was determined by either: ICP-OES or method B which consisted of a summation of strontium quantified by gamma spectroscopy and ICP-MS. The summation method agreed within 10% to the ICP-OES for the total mass of strontium and the subsequent specific activities were determined to be 0.25–0.52 TBq mg-1. Method B was used to determine the isotope abundances by weight% of the purified Sr-82, and the abundances were: Sr-82 (10–20.7%), Sr-83 (0–0.05%), Sr-84 (35–48.5%), Sr-85 (16–25%), Sr-86 (12.5–23%), Sr-87 (0%), and Sr-88 (0–10%). The purified strontium contained mass amounts of Sr-82, Sr-84, Sr-85, Sr-86, and Sr-88 in abundances not associated with natural abundance, and 90% of the strontium was produced by the proton irradiation. A comparison of ICP-OES and method B for the analysis of Sr-82 indicated analysis by ICP-OES would be easier to determine total mass of strontium and comply with regulatory requirements. An ICP-OES analytical method for Sr-82 analysis was established and validated according to regulatory guidelines.

  20. Opioid binding properties of the purified kappa receptor from human placenta

    SciTech Connect

    Ahmed, M.S.; Zhou, D.; Cavinato, A.G.; Maulik, D.

    1989-01-01

    A glycoprotein with a molecular weight of 63,000 has been purified, in an active form, from human placental villus tissue membranes. The binding properties of this glycoprotein to opioid alkaloids and peptides indicates that it is the kappa opiate receptor of human placenta. The receptor binds the tritiated ligands etorphine, bremazocine, ethylketocyclazocine and naloxone specifically and reversibly with Kd values of 3.3, 4.4, 5.1 and 7.0nM, respectively. The binding of /sup 3/H-Bremazocine to the purified receptor is inhibited by the following compounds with the corresponding Ki values EKC, 1.3 x 10/sup -8/M; Dynorphin 1-8, 3.03 x 10/sup -7/; U50,488H, 4.48 x 10/sup -9/; U69-593,2.28 x 10/sup -8/, morphine, 4.05 x 10/sup -6/ DADLE, 6.47 x 10/sup -6/ and naloxone, 2.64 x 10/sup -8/. The purified receptor binds 8 nmole of /sup 3/H-Etorphine and 1.7 nmole /sup 3/H-BZC per mg protein. The theoretical binding capacity of a protein of this molecular weight is 15.8. Although the iodinated purified receptor appears by autoradiography as one band on SDS-PAGE, yet homogeneity of the preparation is not claimed.

  1. Biologic activity of purified cotton bract extracts in man and guinea pig.

    PubMed Central

    Buck, M G; Schachter, E N; Fick, R B; Merrill, W W; Cooper, J A; Keirns, J J; Oliver, J; Wall, J H

    1986-01-01

    Purified aqueous extracts of cotton bract induce acute airway constriction in healthy volunteers never before exposed to cotton bract. The response is similar to that of textile workers who inhale cotton dust. Approximately 60% of volunteers respond to bract extract with significant decreases in lung function, and these volunteers show an increased number of lymphocytes present in their lungs. Following inhalation of bract, the percent of polymorphonuclear leukocytes increases. Macrophages obtained by bronchoalveolar lavage from volunteers pre-challenged with bract extract release increased amounts of chemotactic factor and superoxide anion. Efforts to detect release of histamine and leukotrienes in volunteers following challenge with bract show no increase in urinary histamine and no significant release of leukotrienes in lung lavage fluid. Purified extracts exhibit chemotactic activity in vitro. They also contract guinea pig ileal longitudinal muscle in vitro. This preparation contains mast cells but no basophils, and the H-1 blocker, mepyramine blocks the contraction. Purified bract extracts contain no histamine or endotoxin but other contractors of smooth muscle may be present. The purified extract exhibits spectral, fluorescent, and radioimmune assay properties similar to a leukotriene B-like component. Cotton bract appears to have direct as well as cell-mediated activities. PMID:3011395

  2. Inability of pyrogenic, purified Bordetella pertussis lipid A to induce interleukin-1 release by human monocytes.

    PubMed Central

    Caroff, M; Cavaillon, J M; Fitting, C; Haeffner-Cavaillon, N

    1986-01-01

    Free lipid A of Bordetella pertussis, Neisseria meningitidis, and Escherichia coli lipopolysaccharide (LPS) was prepared by hydrolysis in acetate buffer (pH 4.5); in addition, lipid A from B. pertussis and E. coli was prepared by hydrolysis in mineral acid (HCl). The precipitates obtained were purified by extraction methods in toluene-methanol and are referred to as crude lipid A. Purified lipid A from N. meningitidis and B. pertussis was obtained by extraction in a mixture of chloroform-methanol-water-triethylamine. The different preparations were tested for their pyrogenicity (endogenous pyrogen; EP) and their capacity to trigger the release of interleukin-1 (IL-1; previously known as lymphocyte-activating factor; LAF) by human monocytes. Crude lipid A from E. coli and N. meningitidis were both IL-1 inducers. Crude B. pertussis lipid A (acetate buffer; pH 4.5), which contains a beta-1-6-linked D-glucosamine disaccharide, two phosphoryl groups, and five fatty acids, was pyrogenic and an IL-1 inducer (EP+/LAF+); but crude B. pertussis lipid A (0.25 N HCl), which lacked the glycosidic phosphoryl group, was 1,000-fold less pyrogenic than the diphosphorylated lipid A, yet it retained its IL-1-inducing capacity (EP-/LAF+). Purified N. meningitidis lipid A was not an inducer of IL-1 release and purified B. pertussis lipid A exhibited identical pyrogenicity as the parent LPS but was devoid of any IL-1-release inducing capacity (EP+/LAF-). These results demonstrate that for some endotoxins, purified lipid A is unable to induce IL-1 release by human monocytes; however, it is pyrogenic, supporting the hypothesis that IL-1 and EP are induced by different determinants on the LPS molecule. Images PMID:2876960

  3. 42 CFR 84.1143 - Dust, fume, and mist air-purifying filter tests; performance requirements; general.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... HEALTH AND HUMAN SERVICES OCCUPATIONAL SAFETY AND HEALTH RESEARCH AND RELATED ACTIVITIES APPROVAL OF RESPIRATORY PROTECTIVE DEVICES Dust, Fume, and Mist; Pesticide; Paint Spray; Powered Air-Purifying...

  4. 42 CFR 84.1143 - Dust, fume, and mist air-purifying filter tests; performance requirements; general.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... HEALTH AND HUMAN SERVICES OCCUPATIONAL SAFETY AND HEALTH RESEARCH AND RELATED ACTIVITIES APPROVAL OF RESPIRATORY PROTECTIVE DEVICES Dust, Fume, and Mist; Pesticide; Paint Spray; Powered Air-Purifying...

  5. 42 CFR 84.1143 - Dust, fume, and mist air-purifying filter tests; performance requirements; general.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... HEALTH AND HUMAN SERVICES OCCUPATIONAL SAFETY AND HEALTH RESEARCH AND RELATED ACTIVITIES APPROVAL OF RESPIRATORY PROTECTIVE DEVICES Dust, Fume, and Mist; Pesticide; Paint Spray; Powered Air-Purifying...

  6. 42 CFR 84.1143 - Dust, fume, and mist air-purifying filter tests; performance requirements; general.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... HEALTH AND HUMAN SERVICES OCCUPATIONAL SAFETY AND HEALTH RESEARCH AND RELATED ACTIVITIES APPROVAL OF RESPIRATORY PROTECTIVE DEVICES Dust, Fume, and Mist; Pesticide; Paint Spray; Powered Air-Purifying...

  7. Antibody response of patients after postexposure rabies vaccination with small intradermal doses of purified chick embryo cell vaccine or purified Vero cell rabies vaccine.

    PubMed Central

    Briggs, D. J.; Banzhoff, A.; Nicolay, U.; Sirikwin, S.; Dumavibhat, B.; Tongswas, S.; Wasi, C.

    2000-01-01

    Although the introduction of tissue culture vaccines for rabies has dramatically improved the immunogenicity and safety of rabies vaccines, they are often prohibitively expensive for developing countries. To examine whether smaller doses of these vaccines could be used, we tested the safety and immunogenicity of purified chick embryo cell vaccine (PCECV) on 211 patients in Thailand with World Health Organization (WHO) category II and III exposures to rabies. The patients presented at two Thai hospitals and were randomized into three groups. Patients in Group 1 received 0.1 ml PCECV intradermally at two sites on days 0, 3, 7, and at one site on days 30 and 90. Group 2 was treated similarly, except that purified Vero cell rabies vaccine (PVRV) was used instead of PCECV. Group 3 received 1.0 ml PCECV intramuscularly on days 0, 3, 7, 14, 30 and 90. After 0, 3, 7, 14, 30 and 90 days serum was collected from the subjects and the geometric mean titres (GMTs) of rabies virus neutralizing antibody determined. After 14 days the GMT of 59 patients vaccinated intradermally with PCECV was equivalent to that of patients who received PVRV. Adverse reactions were more frequent in patients who received vaccines intradermally, indicating the reactions were associated with the route of injection, rather than the vaccine per se. We conclude that PCECV is a safe and highly immunogenic vaccine for postexposure rabies vaccination when administered intradermally in 0.1-ml doses using the two-site method ("2,2,2,0,1,1") recommended by WHO. PMID:10859864

  8. Hypoglycemic effect of cooked Lupinus mutabilis and its purified alkaloids in subjects with type-2 diabetes.

    PubMed

    Baldeón, M E; Castro, J; Villacrés, E; Narváez, L; Fornasini, M

    2012-01-01

    Developing countries are experiencing an epidemic of chronic non-communicable chronic diseases with high socio-economic costs. Studies of traditional foods with beneficial health properties could contribute to diminish these problems. Legumes rich in proteins like Lupinus mutabilis decreases blood glucose and improves insulin sensitivity in animals and humans. We report the results of a phase II clinical trial conducted to assess the role of cooked L. mutabilis and its purified alkaloids on blood glucose and insulin in volunteers with diabetes. Results indicate that consumption of cooked L. mutabilis or its purified alkaloids decreased blood glucose and insulin levels. The decreases in serum glucose concentrations from base line to 90 minutes were statistically significant within both treatment groups; however, there were not differences between groups. Serum insulin levels were also decreased in both groups however the differences were not statistically significant. None of the volunteers in either group presented side effects.

  9. In vitro and in vivo antioxidant activities of polysaccharide purified from aloe vera (Aloe barbadensis) gel.

    PubMed

    Kang, Min-Cheol; Kim, Seo Young; Kim, Yoon Taek; Kim, Eun-A; Lee, Seung-Hong; Ko, Seok-Chun; Wijesinghe, W A J P; Samarakoon, Kalpa W; Kim, Young-Sun; Cho, Jin Hun; Jang, Hyeang-Su; Jeon, You-Jin

    2014-01-01

    The in vitro and in vivo antioxidant potentials of a polysaccharide isolated from aloe vera gel were investigated. Enzymatic extracts were prepared from aloe vera gel by using ten digestive enzymes including five carbohydrases and five proteases. Among them, the highest yield was obtained with the Viscozyme extract and the same extract showed the best radical scavenging activity. An active polysaccharide was purified from the Viscozyme extract using ethanol-added separation and anion exchange chromatography. Purified aloe vera polysaccharide (APS) strongly scavenged radicals including DPPH, hydroxyl and alkyl radicals. In addition, APS showed a protective effect against AAPH-induced oxidative stress and cell death in Vero cells as well as in the in vivo zebrafish model. In this study, it is proved that both the in vitro and in vivo antioxidant potentials of APS could be further utilized in relevant industrial applications.

  10. Electron paramagnetic resonance investigation of purified catalyst-free single-walled carbon nanotubes.

    PubMed

    Zaka, Mujtaba; Ito, Yasuhiro; Wang, Huiliang; Yan, Wenjing; Robertson, Alex; Wu, Yimin A; Rümmeli, Mark H; Staunton, David; Hashimoto, Takeshi; Morton, John J L; Ardavan, Arzhang; Briggs, G Andrew D; Warner, Jamie H

    2010-12-28

    Electron paramagnetic resonance of single-walled carbon nanotubes (SWCNTs) has been bedevilled by the presence of paramagnetic impurities. To address this, SWCNTs produced by laser ablation with a nonmagnetic PtRhRe catalyst were purified through a multiple step centrifugation process in order to remove amorphous carbon and catalyst impurities. Centrifugation of a SWCNT solution resulted in sedimentation of carbon nanotube bundles containing clusters of catalyst particles, while isolated nanotubes with reduced catalyst particle content remained in the supernatant. Further ultracentrifugation resulted in highly purified SWCNT samples with a narrow diameter distribution and almost no detectable catalyst particles. Electron paramagnetic resonance (EPR) signals were detected only for samples which contained catalyst particles, with the ultracentrifuged SWCNTs showing no EPR signal at X-band (9.4 GHz) and fields < 0.4 T.

  11. Determining inhibition effects of some aromatic compounds on peroxidase enzyme purified from white and red cabbage

    NASA Astrophysics Data System (ADS)

    Öztekin, Aykut; Almaz, Züleyha; Özdemir, Hasan

    2016-04-01

    Peroxidases (E.C.1.11.1.7) catalyze the one electron oxidation of wide range of substrates. They are used in synthesis reaction, removal of peroxide from industrial wastes, clinical biochemistry and immunoassays. In this study, the white cabbage (Brassica Oleracea var. capitata f. alba) and red cabbage (Brassica oleracea L. var. capitata f. rubra) peroxidase enzymes were purified for investigation of inhibitory effect of some aromatic compounds on these enzymes. IC50 values and Ki constants were calculated for the molecules of 6-Amino nicotinic hydrazide, 6-Amino-5-bromo nicotinic hydrazide, 2-Amino-5-hydroxy benzohydrazide, 4-Amino-3-hydroxy benzohydrazide on purified enzymes and inhibition type of these molecules were determined. (This research was supported by Ataturk University. Project Number: BAP-2015/98).

  12. Protein Affinity Chromatography with Purified Yeast DNA Polymerase α Detects Proteins that Bind to DNA Polymerase

    NASA Astrophysics Data System (ADS)

    Miles, Jeff; Formosa, Tim

    1992-02-01

    We have overexpressed the POL1 gene of the yeast Saccharomyces cerevisiae and purified the resulting DNA polymerase α polypeptide in an apparently intact form. We attached the purified DNA polymerase covalently to an agarose matrix and used this matrix to chromatograph extracts prepared from yeast cells. At least six proteins bound to the yeast DNA polymerase α matrix that did not bind to a control matrix. We speculate that these proteins might be DNA polymerase α accessory proteins. Consistent with this interpretation, one of the binding proteins, which we have named POB1 (polymerase one binding), is required for normal chromosome transmission. Mutations in this gene cause increased chromosome loss and an abnormal cell morphology, phenotypes that also occur in the presence of mutations in the yeast α or δ polymerase genes. These results suggest that the interactions detected by polymerase affinity chromatography are biologically relevant and may help to illuminate the architecture of the eukaryotic DNA replication machinery.

  13. Aspergillus carbonarius polygalacturonases purified by integrated membrane process and affinity precipitation for apple juice production.

    PubMed

    Nakkeeran, Ekambaram; Umesh-Kumar, Sukumaran; Subramanian, Rangaswamy

    2011-02-01

    Aspergillus carbonarius, when grown by submerged and solid-state fermentation, produces different molecular forms of polygalacturonase (PG; EC 3.2.1.15), among them a 42 kDa PG with a high specific activity of 7000 U/mg protein. When the enzymes were purified by integrated membrane process (IMP) and alginate affinity precipitation (AAP), the two processes concentrated different forms of the enzyme. The AAP process selectively purified and concentrated the high active PG whereas the IMP yielded different PGs and also amylase and protease. Evaluation of the AAP enzyme preparations for apple juice preparation under conditions usually employed commercially demonstrated that the high activity PG did not result in good juice clarity. With IMP processed enzymes, juice yields and clarity were similar to that obtained with commercial PG from A. niger.

  14. Immunization of suckling pigs against enteric enterotoxigenic Escherichia coli infection by vaccinating dams with purified pili.

    PubMed

    Nagy, B; Moon, H W; Isaacson, R E; To, C C; Brinton, C C

    1978-07-01

    Pregnant swine (gilts) were vaccinated parenterally with a suspension of purified pili from the porcine enterotoxigenic Escherichia coli strain 987 (09:K103::NM). Gilts injected with placebo served as controls. Suckling pigs born to gilts in both groups were challenged intragastrically with virulent strain 987. The percentage of deaths, incidence and duration of diarrhea, numbers of E. coli in the ilea, and E. coli attachment to the villous epithelia were significantly less in suckling pigs of vaccinated gilts than in those of controls. These results are consistent with the hypothesis that pili of some enterotoxigenic E. coli facilitate adhesion to intestinal epithelia. Vaccination of dams with pili appears to be a means of immunizing against diarrheal disease caused by enterotoxigenic E. coli in suckling neonates. This work confirms the role of somatic pili as colonization and virulence factors and provides another example of safe and effective purified pilus vaccines.

  15. Purifying and Testing Gecko Skin Compounds, a Promising Attractant for Small Brown Treesnakes

    DTIC Science & Technology

    2012-03-13

    that is the case, applying the scent to a more natural- looking (i.e., lizard -like) object may render a stronger and more uniform (across snake ...to other islands. Attractants are particularly needed for <>mall snakes . which are resistant to current control techniques. We extracted skin...compounds from gcckos ( Hemidactylus frcnatus). a preferred prey of the small snakes . and used bioassays to identify which of the successively more purified

  16. New method for purifying histidine-rich glycoprotein from human plasma redefines its functional properties.

    PubMed

    Patel, Kruti K; Poon, Ivan K H; Talbo, Gert H; Perugini, Matthew A; Taylor, Nicole L; Ralph, Troy J; Hoogenraad, Nicholas J; Hulett, Mark D

    2013-06-01

    Histidine-rich glycoprotein (HRG) is a relatively abundant plasma protein that has been implicated in multiple biological processes including immunity, tumor progression, and vascular biology. However, current protocols for purifying HRG from plasma result in the copurification of contaminating proteins and raise questions over the validity of biological activities ascribed to HRG. In this study, we describe a two-step protocol for the large-scale purification of HRG from human plasma using a combination of metal affinity and ion exchange chromatography. The protocol employs a rapid and simple strategy to isolate highly purified HRG that minimizes proteolytic cleavage of the protein. The purification of HRG was assessed at each stage by measuring the amount of HRG immunoreactive protein using a specific monoclonal antibody against total protein, and demonstrated ~1,000-fold purification with an overall yield of ~32%. Mass spectrometry analysis demonstrated that plasma-derived HRG was free of contaminating proteins and gel electrophoresis showed it to have minimal proteolytic degradation. Characterization of protein by physical method showed that the protein exists as a single, monodisperse species. In contrast to the previous studies of HRG purified by different methods, HRG purified using the new procedure demonstrated a reduced profile of functions. Although the HRG retained binding to heparin and phosphatidic acid, it did not interact with necrotic cells or other cellular lipids. These data demonstrate that HRG does not exhibit the broad interactive properties that have been reported previously, suggesting that copurification of HRG-binding partners or other impurities are responsible for some of the reported functional properties. The findings in this study demonstrate that the new purification procedure can provide a ready source of pure HRG to assess ligand specificity and biological function of this important plasma protein.

  17. Human Antibodies against a Purified Glucosylceramide from Cryptococcus neoformans Inhibit Cell Budding and Fungal Growth

    PubMed Central

    Rodrigues, Marcio L.; Travassos, Luiz R.; Miranda, Kildare R.; Franzen, Anderson J.; Rozental, Sonia; de Souza, Wanderley; Alviano, Celuta S.; Barreto-Bergter, Eliana

    2000-01-01

    A major ceramide monohexoside (CMH) was purified from lipidic extracts of Cryptococcus neoformans. This molecule was analyzed by high-performance thin-layer chromatography (HPTLC), gas chromatography coupled with mass spectrometry, and fast atom bombardment-mass spectrometry. The cryptococcal CMH is a β-glucosylceramide, with the carbohydrate residue attached to 9-methyl-4,8-sphingadienine in amidic linkage to 2-hydroxyoctadecanoic acid. Sera from patients with cryptococcosis and a few other mycoses reacted with the cryptococcal CMH. Specific antibodies were purified from patients' sera by immunoadsorption on the purified glycolipid followed by protein G affinity chromatography. The purified antibodies to CMH (mainly immunoglobulin G1) bound to different strains and serological types of C. neoformans, as shown by flow cytofluorimetry and immunofluorescence labeling. Transmission electron microscopy of yeasts labeled with immunogold-antibodies to CMH and immunostaining of isolated cell wall lipid extracts separated by HPTLC showed that the cryptococcal CMH predominantly localizes to the fungal cell wall. Confocal microscopy revealed that the β-glucosylceramide accumulates mostly at the budding sites of dividing cells with a more disperse distribution at the cell surface of nondividing cells. The increased density of sphingolipid molecules seems to correlate with thickening of the cell wall, hence with its biosynthesis. The addition of human antibodies to CMH to cryptococcal cultures of both acapsular and encapsulated strains of C. neoformans inhibited cell budding and cell growth. This process was complement-independent and reversible upon removal of the antibodies. The present data suggest that the cryptococcal β-glucosylceramide is a fungal antigen that plays a role on the cell wall synthesis and yeast budding and that antibodies raised against this component are inhibitory in vitro. PMID:11083830

  18. Contamination of a purified water system by Aspergillus fumigatus in a new endoscopy reprocessing unit.

    PubMed

    Khalsa, Kamaljit; Smith, Andrew; Morrison, Patrick; Shaw, David; Peat, Marie; Howard, Paul; Hamilton, Kate; Stewart, Alan

    2014-12-01

    Use of purified water for the final rinse stage of disinfected endoscopes is an important element of quality control. We describe the detection and management of Aspergillus fumigatus contamination of a new reverse osmosis unit supplying 10 automated endoscope reprocessor basins. Prompt detection and reaction to this contaminant were possible because of the introduction of a comprehensive program for microbiological monitoring of rinse waters, which included total viable counts, endotoxin, conductivity, and Pseudomonas spp.

  19. Purifying Selection and Molecular Adaptation in the Genome of Verminephrobacter, the Heritable Symbiotic Bacteria of Earthworms

    PubMed Central

    Kjeldsen, Kasper U.; Bataillon, Thomas; Pinel, Nicolás; De Mita, Stéphane; Lund, Marie B.; Panitz, Frank; Bendixen, Christian; Stahl, David A.; Schramm, Andreas

    2012-01-01

    While genomic erosion is common among intracellular symbionts, patterns of genome evolution in heritable extracellular endosymbionts remain elusive. We study vertically transmitted extracellular endosymbionts (Verminephrobacter, Betaproteobacteria) that form a beneficial, species-specific, and evolutionarily old (60–130 Myr) association with earthworms. We assembled a draft genome of Verminephrobacter aporrectodeae and compared it with the genomes of Verminephrobacter eiseniae and two nonsymbiotic close relatives (Acidovorax). Similar to V. eiseniae, the V. aporrectodeae genome was not markedly reduced in size and showed no A–T bias. We characterized the strength of purifying selection (ω = dN/dS) and codon usage bias in 876 orthologous genes. Symbiont genomes exhibited strong purifying selection (ω = 0.09 ± 0.07), although transition to symbiosis entailed relaxation of purifying selection as evidenced by 50% higher ω values and less codon usage bias in symbiont compared with reference genomes. Relaxation was not evenly distributed among functional gene categories but was overrepresented in genes involved in signal transduction and cell envelope biogenesis. The same gene categories also harbored instances of positive selection in the Verminephrobacter clade. In total, positive selection was detected in 89 genes, including also genes involved in DNA metabolism, tRNA modification, and TonB-dependent iron uptake, potentially highlighting functions important in symbiosis. Our results suggest that the transition to symbiosis was accompanied by molecular adaptation, while purifying selection was only moderately relaxed, despite the evolutionary age and stability of the host association. We hypothesize that biparental transmission of symbionts and rare genetic mixing during transmission can prevent genome erosion in heritable symbionts. PMID:22333491

  20. The Experience of Implementation of Innovative Technology of Quarry Waste Water Purifying in Kuzbass Open Pit

    NASA Astrophysics Data System (ADS)

    Lesin, Yu V.; Hellmer, M. C.

    2016-08-01

    Among all industries in Kuzbass (Western Siberia, Russia) the coal industry provides the most environmental threat. However, the construction of new and maintenance of existing open pit mines do not often correspond to the tasks of improving the environmental safety of surface mining. So the article describes the use of innovative quarry waste water purifying technology implemented in Kuzbass open pit mine «Shestaki». This technology is based on using artificial filter arrays made of overburden rock.

  1. Myogenic Growth Factor Present in Skeletal Muscle is Purified by Heparin-Affinity Chromatography

    NASA Astrophysics Data System (ADS)

    Kardami, Elissavet; Spector, Dennis; Strohman, Richard C.

    1985-12-01

    A myogenic growth factor has been purified from a skeletal muscle, the anterior latissimus dorsi, of adult chickens. In the range of 1-10 ng, this factor stimulates DNA synthesis as well as protein and muscle-specific myosin accumulation in myogenic cell cultures. Purification is achieved through binding of the factor to heparin. The factor is distinct from transferrin and works synergistically with transferrin in stimulating myogenesis in vitro.

  2. PNA lectin for purifying mouse acinar cells from the inflamed pancreas.

    PubMed

    Xiao, Xiangwei; Fischbach, Shane; Fusco, Joseph; Zimmerman, Ray; Song, Zewen; Nebres, Philip; Ricks, David Matthew; Prasadan, Krishna; Shiota, Chiyo; Husain, Sohail Z; Gittes, George K

    2016-02-17

    Better methods for purifying human or mouse acinar cells without the need for genetic modification are needed. Such techniques would be advantageous for the specific study of certain mechanisms, such as acinar-to-beta-cell reprogramming and pancreatitis. Ulex Europaeus Agglutinin I (UEA-I) lectin has been used to label and isolate acinar cells from the pancreas. However, the purity of the UEA-I-positive cell fraction has not been fully evaluated. Here, we screened 20 widely used lectins for their binding specificity for major pancreatic cell types, and found that UEA-I and Peanut agglutinin (PNA) have a specific affinity for acinar cells in the mouse pancreas, with minimal affinity for other major pancreatic cell types including endocrine cells, duct cells and endothelial cells. Moreover, PNA-purified acinar cells were less contaminated with mesenchymal and inflammatory cells, compared to UEA-I purified acinar cells. Thus, UEA-I and PNA appear to be excellent lectins for pancreatic acinar cell purification. PNA may be a better choice in situations where mesenchymal cells or inflammatory cells are significantly increased in the pancreas, such as type 1 diabetes, pancreatitis and pancreatic cancer.

  3. PNA lectin for purifying mouse acinar cells from the inflamed pancreas

    PubMed Central

    Xiao, Xiangwei; Fischbach, Shane; Fusco, Joseph; Zimmerman, Ray; Song, Zewen; Nebres, Philip; Ricks, David Matthew; Prasadan, Krishna; Shiota, Chiyo; Husain, Sohail Z.; Gittes, George K.

    2016-01-01

    Better methods for purifying human or mouse acinar cells without the need for genetic modification are needed. Such techniques would be advantageous for the specific study of certain mechanisms, such as acinar-to-beta-cell reprogramming and pancreatitis. Ulex Europaeus Agglutinin I (UEA-I) lectin has been used to label and isolate acinar cells from the pancreas. However, the purity of the UEA-I-positive cell fraction has not been fully evaluated. Here, we screened 20 widely used lectins for their binding specificity for major pancreatic cell types, and found that UEA-I and Peanut agglutinin (PNA) have a specific affinity for acinar cells in the mouse pancreas, with minimal affinity for other major pancreatic cell types including endocrine cells, duct cells and endothelial cells. Moreover, PNA-purified acinar cells were less contaminated with mesenchymal and inflammatory cells, compared to UEA-I purified acinar cells. Thus, UEA-I and PNA appear to be excellent lectins for pancreatic acinar cell purification. PNA may be a better choice in situations where mesenchymal cells or inflammatory cells are significantly increased in the pancreas, such as type 1 diabetes, pancreatitis and pancreatic cancer. PMID:26884345

  4. Pepsin degradation of Cry1A(b) protein purified from genetically modified maize (Zea mays).

    PubMed

    de Luis, Ruth; Lavilla, María; Sánchez, Lourdes; Calvo, Miguel; Pérez, María D

    2010-02-24

    The aim of this work was to study the in vitro digestion of Cry1A(b) protein by pepsin. To perform this work, a protein fraction purified from transgenic maize by immunoadsorption was employed. The undigested fraction showed several bands of molecular weight ranging between 14 and 70 kDa when assayed by SDS-PAGE. These bands were identified as corresponding to Cry1A(b) protein by immunochemical techniques and mass spectrometry. The rate of degradation of the purified fraction by pepsin estimated by ELISA was found to be about 75% within 30 min, and the protein concentration remained constant up to 4 h. In all treated samples, the full-length protein and fragments present in Cry1A(b) fraction were absent and peptides of less than 8.5 kDa were mainly found by SDS-PAGE and mass spectrometry. These peptides did not react with antiserum against Cry1A(b) protein by Western blotting. These results suggest that Cry1A(b) fraction purified from transgenic maize is rapidly and extensively degraded by pepsin, giving peptides of low molecular mass.

  5. Purified eicosapentaenoic acid induces prolonged survival of cardiac allografts and generates regulatory T cells.

    PubMed

    Iwami, D; Zhang, Q; Aramaki, O; Nonomura, K; Shirasugi, N; Niimi, M

    2009-06-01

    Fish oil, which is rich in eicosapentaenoic acid (EPA), has been found to have immunomodulatory effects. We examined whether administration of purified EPA affected survival of fully mismatched murine cardiac allografts. Hearts from C57BL/10 (H-2(b)) mice were transplanted into CBA (H-2(k)) recipients treated with one intraperitoneal dose of purified EPA the day of transplantation. Untreated CBA recipients and recipients given 0.1 g/kg of EPA rejected C57BL/10 hearts (median survival time [MST], 8 and 13 days, respectively). With a 1.0 g/kg dose of EPA, graft survival was markedly prolonged (MST >100 days). To determine whether regulatory cells were generated, naïve mice (secondary recipients) underwent adoptive transfer of splenocytes from EPA-treated primary recipients and cardiac allograft transplantation. Adoptive transfer of whole, CD4(+) and CD4(+)CD25(+) splenocytes from EPA-treated recipients induced indefinite survival in secondary recipients. Flow cytometry showed that the CD4(+)CD25(+) cells were Foxp3(+). In reverse transcriptase-polymerase chain reaction (RT-PCR) studies, the expression of peroxisome proliferator-activated receptor gamma (PPARgamma) mRNA was upregulated by EPA treatment. A PPARgamma antagonist abrogated the prolongation of graft survival induced by EPA treatment (MST, 13 days). Thus, in our model, purified EPA induced prolonged survival of fully mismatched cardiac allografts and generated regulatory T cells dependent on PPARgamma activation.

  6. Photocatalytic air purifiers for indoor air: European standard and pilot room experiments.

    PubMed

    Costarramone, N; Cantau, C; Desauziers, V; Pécheyran, C; Pigot, T; Lacombe, S

    2016-09-15

    At the European level (CEN/TC386), some efforts are currently devoted to new standards for comparing the efficiency of commercial photocatalytic material/devices in various application fields. Concerning prototype or commercial indoor photocatalytic air purifiers designed for volatile organic compounds (VOC) abatement, the methodology is based on a laboratory airtight chamber. The photocatalytic function is demonstrated by the mineralization of a mixture of five VOCs. Experimental data were obtained for four selected commercial devices and three commercial materials: drop of VOC concentration, but also identification of secondary species (with special attention to formaldehyde), mineralization rates, and Clean Air Delivery Rate (CADR). With two efficient air purifiers, these laboratory experiments were compared to the results in two experimental rooms (35-40 m(3)) where air pollution was introduced through wooden floor and furniture. The systems' ageing was also studied. The safety of the commercial products was also assessed by the determination of nanoparticle release. Standardized tests are useful to rank photocatalytic air purifiers and passive materials and to discard inefficient ones. A good correlation between the standard experiments and the experimental room experiments was found, even if in the latter case, the concentration of lower weight VOCs drops less quickly than that of heavier VOCs.

  7. Relaxation of purifying selection on the SAD lineage of live attenuated oral vaccines for rabies virus.

    PubMed

    Hughes, Austin L

    2009-09-01

    Analysis of patterns of nucleotide sequence diversity in wild-type rabies virus (RABV) genomes and in the SAD live attenuated oral vaccine lineage was used to test for the relaxation of purifying selection in the latter and provide evidence regarding the genomic regions where such relaxation of selection occurs. The wild-type sequences showed evidence of strong past and ongoing purifying selection both on nonsynonymous sites in coding regions and on non-coding regions, particularly the start, end and 5' UTR regions. SAD vaccine sequences showed a relaxation of purifying selection at nonsynonymous sites in coding regions, resulting a substantial number of amino acid sequence polymorphisms at sites that were invariant in the wild-type sequences. Moreover, SAD vaccine sequences showed high levels of mutation accumulation in the non-coding regions that were most conserved in the wild-type sequences. Understanding the biological effects of the unique mutations accumulated in the vaccine lineage is important because of their potential effects on antigenicity and effectiveness of the vaccine.

  8. Immunodiagnosis of human cysticercosis (Taenia solium) with antigens purified by monoclonal antibodies.

    PubMed Central

    Nascimento, E; Tavares, C A; Lopes, J D

    1987-01-01

    Monoclonal antibodies were generated from mice immunized with scolex protein antigen of Cysticercus cellulosae. Three monoclonal antibodies specific for cysticercal antigens, which did not show any cross-reactivity with Taenia solium or Taenia saginata antigens, were selected. Each monoclonal antibody coupled to Sepharose could purify one antigen, which appeared as a single band on polyacrylamide gel electrophoresis. When antigens purified by monoclonal antibodies were used to detect antibody in serum samples taken from patients with cysticercosis, taeniasis, and other parasitic infections in an enzyme-linked immunosorbent assay, cross-reactivity was observed until a serum dilution of 1:128 was reached. Since serum samples from unexposed subjects showed positive reactions until a dilution of 1:64 was reached, we chose a discriminative dilution (1:128) above which no cross-reaction was observed. The percent positive serum samples from cysticercosis patients was 100% by the enzyme-linked immunosorbent assay with any of the antigens purified by monoclonal antibodies. Images PMID:3611310

  9. Pseudomonas aeruginosa arylsulfatase: a purified enzyme for the mild hydrolysis of steroid sulfates.

    PubMed

    Stevenson, Bradley J; Waller, Christopher C; Ma, Paul; Li, Kunkun; Cawley, Adam T; Ollis, David L; McLeod, Malcolm D

    2015-10-01

    The hydrolysis of sulfate ester conjugates is frequently required prior to analysis for a range of analytical techniques including gas chromatography-mass spectrometry (GC-MS). Sulfate hydrolysis may be achieved with commercial crude arylsulfatase enzyme preparations such as that derived from Helix pomatia but these contain additional enzyme activities such as glucuronidase, oxidase, and reductase that make them unsuitable for many analytical applications. Strong acid can also be used to hydrolyze sulfate esters but this can lead to analyte degradation or increased matrix interference. In this work, the heterologously expressed and purified arylsulfatase from Pseudomonas aeruginosa is shown to promote the mild enzyme-catalyzed hydrolysis of a range of steroid sulfates. The substrate scope of this P. aeruginosa arylsulfatase hydrolysis is compared with commercial crude enzyme preparations such as that derived from H. pomatia. A detailed kinetic comparison is reported for selected examples. Hydrolysis in a urine matrix is demonstrated for dehydroepiandrosterone 3-sulfate and epiandrosterone 3-sulfate. The purified P. aeruginosa arylsulfatase contains only sulfatase activity allowing for the selective hydrolysis of sulfate esters in the presence of glucuronide conjugates as demonstrated in the short three-step chemoenzymatic synthesis of 5α-androstane-3β,17β-diol 17-glucuronide (ADG, 1) from epiandrosterone 3-sulfate. The P. aeruginosa arylsulfatase is readily expressed and purified (0.9 g per L of culture) and thus provides a new and selective method for the hydrolysis of steroid sulfate esters in analytical sample preparation.

  10. Inference of purifying and positive selection in three subspecies of chimpanzees (Pan troglodytes) from exome sequencing.

    PubMed

    Bataillon, Thomas; Duan, Jinjie; Hvilsom, Christina; Jin, Xin; Li, Yingrui; Skov, Laurits; Glemin, Sylvain; Munch, Kasper; Jiang, Tao; Qian, Yu; Hobolth, Asger; Wang, Jun; Mailund, Thomas; Siegismund, Hans R; Schierup, Mikkel H

    2015-03-30

    We study genome-wide nucleotide diversity in three subspecies of extant chimpanzees using exome capture. After strict filtering, Single Nucleotide Polymorphisms and indels were called and genotyped for greater than 50% of exons at a mean coverage of 35× per individual. Central chimpanzees (Pan troglodytes troglodytes) are the most polymorphic (nucleotide diversity, θw = 0.0023 per site) followed by Eastern (P. t. schweinfurthii) chimpanzees (θw = 0.0016) and Western (P. t. verus) chimpanzees (θw = 0.0008). A demographic scenario of divergence without gene flow fits the patterns of autosomal synonymous nucleotide diversity well except for a signal of recent gene flow from Western into Eastern chimpanzees. The striking contrast in X-linked versus autosomal polymorphism and divergence previously reported in Central chimpanzees is also found in Eastern and Western chimpanzees. We show that the direction of selection statistic exhibits a strong nonmonotonic relationship with the strength of purifying selection S, making it inappropriate for estimating S. We instead use counts in synonymous versus nonsynonymous frequency classes to infer the distribution of S coefficients acting on nonsynonymous mutations in each subspecies. The strength of purifying selection we infer is congruent with the differences in effective sizes of each subspecies: Central chimpanzees are undergoing the strongest purifying selection followed by Eastern and Western chimpanzees. Coding indels show stronger selection against indels changing the reading frame than observed in human populations.

  11. Purified polysaccharides of Geoffroea spinosa barks have anticoagulant and antithrombotic activities devoid of hemorrhagic risks.

    PubMed

    Souza, Racquel O S; Assreuy, Ana M S; Madeira, Juliana C; Chagas, Francisco D S; Parreiras, Luane A; Santos, Gustavo R C; Mourão, Paulo A S; Pereira, Maria G

    2015-06-25

    Polysaccharides were extracted from the barks of Geoffroea spinosa, purified using anion exchange chromatography and characterized by chemical and methylation analysis, complemented by infrared and NMR spectroscopies. These polysaccharides were tested for their anticoagulant, antithrombotic and antiplatelet activities and also for their effects on bleeding. Unfractionated polysaccharide contains low levels of protein and high levels of carbohydrate (including hexuronic acid). The purified polysaccharides (fractions FII and FIII) are composed of arabinose (Ara), rhamnose (Rha), hexuronic acid, small amounts of galactose, but no sulfate ester. They have highly complex structure, which was partially characterized. NMR and methylation analysis indicate that the polysaccharides have a core of α-Rhap and branches of 5-linked α-Araf. Residues of 4-linked α-GalpA are also found in the structure. The unfractionated (TPL) and fraction FIII, but not fractions FI and FII, prolonged the activated partial thromboplastin time (aPTT). TPL, FII and FIII inhibited the platelet aggregation induced by ADP. More significantly, both unfractionated and purified fractions exhibited potent antithrombotic effect (31-60%) and the fractions did not modify the bleeding tendency. These plant polysaccharides could be alternative source of new anticoagulant, antiplatelet and antithrombotic compounds devoid of the undesirable risk of hemorrhage.

  12. Population Level Purifying Selection and Gene Expression Shape Subgenome Evolution in Maize.

    PubMed

    Pophaly, Saurabh D; Tellier, Aurélien

    2015-12-01

    The maize ancestor experienced a recent whole-genome duplication (WGD) followed by gene erosion which generated two subgenomes, the dominant subgenome (maize1) experiencing fewer deletions than maize2. We take advantage of available extensive polymorphism and gene expression data in maize to study purifying selection and gene expression divergence between WGD retained paralog pairs. We first report a strong correlation in nucleotide diversity between duplicate pairs, except for upstream regions. We then show that maize1 genes are under stronger purifying selection than maize2. WGD retained genes have higher gene dosage and biased Gene Ontologies consistent with previous studies. The relative gene expression of paralogs across tissues demonstrates that 98% of duplicate pairs have either subfunctionalized in a tissuewise manner or have diverged consistently in their expression thereby preventing functional complementation. Tissuewise subfunctionalization seems to be a hallmark of transcription factors, whereas consistent repression occurs for macromolecular complexes. We show that dominant gene expression is a strong determinant of the strength of purifying selection, explaining the inferred stronger negative selection on maize1 genes. We propose a novel expression-based classification of duplicates which is more robust to explain observed polymorphism patterns than the subgenome location. Finally, upstream regions of repressed genes exhibit an enrichment in transposable elements which indicates a possible mechanism for expression divergence.

  13. Effect of purified diets and phenobarbital withdrawal on the phenotypic stability of altered hepatic foci (AHF)

    SciTech Connect

    Glauert, H.P.; Schwarz, M.; Pitot, H.C.

    1986-03-05

    The effect of the short-term withdrawal of phenobarbital (PB) and of the feeding of purified diets during the long-term withdrawal of PB on the stability of AHF were studied. In both experiments, female CD rats initially received an intragastric dose of diethylnitrosamine (10 mg/kg) 20 hours after being subjected to partial hepatectomy. In the short-term study, rats were fed 0.05% PB in a cereal-based diet for 6 months; at this time, half of the rats were killed whereas the other half were withdrawn from PB for 10 days before sacrifice. Withdrawing PB for 10 days resulted in a decrease in the number and volume of AHF, particularly those which stained positively for gamma-glutamyltranspeptidase (GGT). In the long-term experiment, rats were fed 0.05% PB in a cereal-based diet containing PB and fed either a low-fat or a high-fat purified diet without PB for 8 months. At this time, the number and volume of AHF were much less than that seen at the time of PB withdrawal, and the distribution of phenotypes was altered: the percentage of foci containing GGT as a marker decreased dramatically. These results indicate that the observable number and total volume of AHF rapidly decrease after the withdrawal of PB from rats fed a cereal-based diet and that the feeding of purified diets after such PB withdrawal does not result in the reappearance of AHF.

  14. Purified E255L Mutant SERCA1a and Purified PfATP6 Are Sensitive to SERCA-type Inhibitors but Insensitive to Artemisinins*

    PubMed Central

    Cardi, Delphine; Pozza, Alexandre; Arnou, Bertrand; Marchal, Estelle; Clausen, Johannes D.; Andersen, Jens Peter; Krishna, Sanjeev; Møller, Jesper V.; le Maire, Marc; Jaxel, Christine

    2010-01-01

    The antimalarial drugs artemisinins have been described as inhibiting Ca2+-ATPase activity of PfATP6 (Plasmodium falciparum ATP6) after expression in Xenopus oocytes. Mutation of an amino acid residue in mammalian SERCA1 (Glu255) to the equivalent one predicted in PfATP6 (Leu) was reported to induce sensitivity to artemisinin in the oocyte system. However, in the present experiments, we found that artemisinin did not inhibit mammalian SERCA1a E255L either when expressed in COS cells or after purification of the mutant expressed in Saccharomyces cerevisiae. Moreover, we found that PfATP6 after expression and purification from S. cerevisiae was insensitive to artemisinin and significantly less sensitive to thapsigargin and 2,5-di(tert-butyl)-1,4-benzohydroquinone than rabbit SERCA1 but retained higher sensitivity to cyclopiazonic acid, another type of SERCA1 inhibitor. Although mammalian SERCA and purified PfATP6 appear to have different pharmacological profiles, their insensitivity to artemisinins suggests that the mechanism of action of this class of drugs on the calcium metabolism in the intact cell is complex and cannot be ascribed to direct inhibition of PfATP6. Furthermore, the successful purification of PfATP6 affords the opportunity to develop new antimalarials by screening for inhibitors against PfATP6. PMID:20530490

  15. Preparation and Thermal Properties of High-Purified Molten Nitrate Salt Materials with Heat Transfer and Storage

    NASA Astrophysics Data System (ADS)

    Zhang, Hongtao; Zhao, Youjing; Li, Jingli; Shi, Lijie; Wang, Min

    2015-12-01

    This paper focuses on thermal stability of molten salts, operating temperature range and latent heat of molten salts at a high temperature. In this experiment, multi-component molten salts (purified Solar Salt) composed of purified NaNO3 and purified KNO3 were prepared by statical mixing method. Compared with unpurified Solar Salt, purified Solar Salt had a higher thermal stability. The optimal temperature would be increased from 500°C to 550°C, and the upper limitation temperature increases from 577.5°C to 593.7°C. Meanwhile, thermal stability and thermal cycling analysis showed the purified Solar Salt had a lower melting point and the deterioration time of molten salts was reduced. The melting point of purified Solar Salt decreases sharply to 223.8°C and the latent heat increases from 74.39 J/g to 80.79 J/g. Besides, the XRD and chemical analysis test indicated that the degree of degradation reduced and the thermal storage efficiency of purified Solar Salt was improved.

  16. A purified Bacillus thuringiensis crystal protein with therapeutic activity against the hookworm parasite Ancylostoma ceylanicum

    PubMed Central

    Cappello, Michael; Bungiro, Richard D.; Harrison, Lisa M.; Bischof, Larry J.; Griffitts, Joel S.; Barrows, Brad D.; Aroian, Raffi V.

    2006-01-01

    Crystal (Cry) proteins produced by the soil bacterium Bacillus thuringiensis (Bt) are harmless to vertebrates, but they are highly toxic to insects and nematodes. Their value in controlling insects that destroy crops and transmit human diseases is well established. Although it has recently been demonstrated that a few individual Bt Cry proteins, such as Cry5B, are toxic to a wide range of free-living nematodes, the potential activity of purified Cry proteins against parasitic nematodes remains largely unknown. We report here studies aimed at characterizing in vitro and in vivo anthelminthic activities of purified recombinant Cry5B against the hookworm parasite Ancylostoma ceylanicum, a bloodfeeding gastrointestinal nematode for which humans are permissive hosts. By using in vitro larval development assays, Cry5B was found to be highly toxic to early stage hookworm larvae. Exposure of adult A. ceylanicum to Cry5B was also associated with significant toxicity, including a substantial reduction in egg excretion by adult female worms. To demonstrate therapeutic efficacy in vivo, hamsters infected with A. ceylanicum were treated with three daily oral doses of purified Cry5B, the benzimidazole anthelminthic mebendazole, or buffer. Compared with control (buffer-treated) animals, infected hamsters that received Cry5B showed statistically significant improvements in growth and blood hemoglobin levels as well as reduced worm burdens that were comparable to the mebendazole-treated animals. These data demonstrate that Cry5B is highly active in vitro and in vivo against a globally significant nematode parasite and that Cry5B warrants further clinical development for human and veterinary use. PMID:17005719

  17. Liquid-vapor interfacial tension of blood plasma, serum and purified protein constituents thereof.

    PubMed

    Krishnan, Anandi; Wilson, Arwen; Sturgeon, Jacqueline; Siedlecki, Christopher A; Vogler, Erwin A

    2005-06-01

    A systematic study of water-air (liquid-vapor, LV) interfacial tension gamma(lv) of blood plasma and serum derived from four different mammalian species (human, bovine, ovine and equine) reveals nearly identical concentration-dependence (dgamma(lv)/dlnC(B); where C(B) is plasma/serum dilution expressed in v/v concentration units). Comparison of results to a previously-published survey of purified human-blood proteins further reveals that dgamma(lv)/dlnC(B) of plasma and serum is surprisingly similar to that of purified protein constituents. It is thus concluded that any combination of blood-protein constituents will be substantially similar because dgamma(lv)/dlnC(B) of individual proteins are very similar. Experimental results are further interpreted in terms of a recently-developed theory emphasizing the controlling role of water in protein adsorption. Accordingly, the LV interphase saturates with protein adsorbed from bulk solution at a fixed weight-volume concentration ( approximately 436 mg/mL) independent of protein identity or mixture. As a direct consequence, dgamma(lv)/dlnC(B) of purified proteins closely resembles that of mixed solutions and does not depend on the relative proportions of individual proteins comprising a mixture. Thus variations in the plasma proteome between species are not reflected in dgamma(lv)/dlnC(B) nor is serum different from plasma in this regard, despite being depleted of coagulation proteins (e.g. fibrinogen). A comparison of pendant-drop and Wilhelmy-balance tensiometry as tools for assessing protein gamma(lv) shows that measurement conditions employed in the typical Wilhelmy plate approach fails to achieve the steady-state adsorption state that is accessible to pendant-drop tensiometry.

  18. Speech intelligibility while wearing full-facepiece air-purifying respirators.

    PubMed

    Coyne, Karen M; Barker, Daniel J

    2014-01-01

    Intelligible speech communication while wearing air-purifying respirators is critical for law enforcement officers, particularly when they are communicating with each other or the public. The National Institute for Occupational Safety and Health (NIOSH) requires a 70% overall performance rating to pass speech intelligibility certification for commercial chemical, biological, radiological, and nuclear air-purifying respirators. However, the speech intelligibility of certified respirators is not reported and the impact on operational performance is unknown. The objective of this effort was to assess the speech intelligibility of 12 certified air-purifying respirators and to predict their impact on operational performance. The NIOSH respirator certification standard testing procedures were followed. Regression equations were fit to data from studies that examined the impact of degraded speech intelligibility on operational performance of simple and complex missions. The impact of the tested respirators on operational performance was estimated from these equations. Performance ratings observed for each respirator were: MSA Millennium (90%), 3M FR-M40 (88%), MSA Ultra Elite (87%), Scott M110 (86%), North 5400 (85%), Scott M120 (85%), Avon C50 (84%), Avon FM12 (84%), Survivair Optifit (81%), Drager CDR 4500 (81%), Peltor-AOSafety M-TAC (79%), and 3M FR-7800B (78%). The Millennium and FR-M40 had statistically significantly higher scores than the FR-7800B. The Millennium also scored significantly higher than the M-TAC. All of the tested respirators were predicted to have little impact on simple and complex mission performance times and on simple mission success rate. However, the regression equations showed that 75% of missions that require complex communications would be completed while wearing the Millennium, FR-M40, or Ultra Elite but that only 60% would be completed successfully while wearing the FR-7800B. These results suggest that some certified respirators may have

  19. Can phlorotannins purified extracts constitute a novel pharmacological alternative for microbial infections with associated inflammatory conditions?

    PubMed

    Lopes, Graciliana; Sousa, Carla; Silva, Luís R; Pinto, Eugénia; Andrade, Paula B; Bernardo, João; Mouga, Teresa; Valentão, Patrícia

    2012-01-01

    Bacterial and fungal infections and the emerging multidrug resistance are driving interest in fighting these microorganisms with natural products, which have generally been considered complementary to pharmacological therapies. Phlorotannins are polyphenols restricted to brown seaweeds, recognized for their biological capacity. This study represents the first research on the antibacterial, antifungal, anti-inflammatory and antioxidant activity of phlorotannins purified extracts, which were obtained from ten dominant brown seaweeds of the occidental Portuguese coast.Phlorotannins content was determined by the specific dimethoxybenzaldehyde (DMBA) method and a yield between 75 and 969 mg/Kg phloroglucinol units (dry matter) was obtained. Fucus spiralis ranked first, followed by three Cystoseira species. The anti-inflammatory potential of the purified extracts was assessed via inhibitory effect on nitric oxide (NO) production by lipopolysaccharide-stimulated RAW 264.7 macrophage cells, Cystoseira tamariscifolia being the one showing promising activity for the treatment of inflammation. NO scavenging ability was also addressed in cell free systems, F. spiralis being the species with highest capacity. The antimicrobial potential of the extracts was checked against five Gram-positive and four Gram-negative bacteria and three fungi strains, that commonly colonize skin and mucosa and are responsible for food contamination. The different extracts were more effective against Gram-positive bacteria, Staphylococcus epidermidis being the most susceptible species. Concerning antifungal activity, Trichophyton rubrum was the most sensitive species.Although the molecular mechanisms underlying these properties remain poorly understood, the results obtained turn phlorotannins purified extracts a novel and potent pharmacological alternative for the treatment of a wide range of microbial infections, which usually also present an inflammatory component. In addition to the biological

  20. Subunit Structure Differences in RNA Polymerase II Purified from Ungerminated versus Germinated Wheat Embryos 1

    PubMed Central

    Jendrisak, Jerry; Skuzeski, Jim

    1983-01-01

    DNA-dependent RNA polymerase II (RNAP II) was purified from wheat embryos germinated for 0, 12, 24, and 36 hours and examined with several polyacrylamide gel electrophoretic systems. A changing electrophoretic pattern of RNAP II was observed on nondenaturing polyacrylamide gels. Subunit structure analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicated that from ungerminated embryos, RNAP IIA was almost exclusively obtained which has a subunit structure identical to that established for wheat germ RNAP II previously (Jendrisak, Burgess 1977 Biochemistry 16: 1959-1964). Twelve polypeptides with molecular weights × 10−3 of 220, 140, 42, 40, 27, 25, 21, 20, 17.8, 17.0, 16.3, and 16.0 were routinely found to be associated with the purified enzyme. From embryos germinated for 36 hours, RNAP IIB was almost exclusively obtained which has a largest subunit of 180,000 mol wt instead of 220,000. From embryos germinated for 24 hours, an approximately equimolar mixture of RNAP IIA and IIB was obtained. Peptide maps of the 220,000 and 180,000 mol wt polypeptides of RNAP IIA and IIB were virtually identical, indicative of a precursor-product relationship for the two polypeptides. In addition to these results, SDS-PAGE indicated that the stoichiometry of the 27,000 mol wt polypeptide increased at the expense of the 25,000 mol wt polypeptide during germination and concomitantly with the appearance of the 180,000 molecular weight polypeptide. No modifications (e.g. gain, loss, or altered mobilities on analytical gels) in any of the other RNAP II subunits were observed in enzyme purified from embryos after various times of germination as determined by a variety of electrophoretic analyses under denaturing conditions. Images Fig. 1 Fig. 3 Fig. 4 Fig. 5 PMID:16663122

  1. Purified TMEM16A is sufficient to form Ca2+-activated Cl− channels

    PubMed Central

    Terashima, Hiroyuki; Picollo, Alessandra; Accardi, Alessio

    2013-01-01

    Ca2+-activated Cl− channels (CaCCs) are key regulators of numerous physiological functions, ranging from electrolyte secretion in airway epithelia to cellular excitability in sensory neurons and muscle fibers. Recently, TMEM16A (ANO1) and -B were shown to be critical components of CaCCs. It is still unknown whether they are also sufficient to form functional CaCCs, or whether association with other subunits is required. Recent reports suggest that the Ca2+ sensitivity of TMEM16A is mediated by its association with calmodulin, suggesting that functional CaCCs are heteromultimers. To test whether TMEM16A is necessary and sufficient to form functional CaCCs, we expressed, purified, and reconstituted human TMEM16A. The purified protein mediates Ca2+-dependent Cl− transport with submicromolar sensitivity to Ca2+, consistent with what is seen in patch–clamp experiments. The channel is synergistically gated by Ca2+ and voltage, so that opening is promoted by depolarizing potentials. Mutating two conserved glutamates in the TM6-7 intracellular loop selectively abolishes the Ca2+ dependence of reconstituted TMEM16A, in a manner similar to what was reported for the heterologously expressed channel. Well-characterized CaCC blockers inhibit Cl− transport with Kis comparable to those measured for native and heterologously expressed CaCCs. Finally, direct physical interactions between calmodulin and TMEM16A could not be detected in copurification experiments or in functional assays. Our results demonstrate that purified TMEM16A is necessary and sufficient to recapitulate the biophysical and pharmacological properties of native and heterologously expressed CaCCs. Our results also show that association of TMEM16A with other proteins, such as calmodulin, is not required for function. PMID:24167264

  2. Protective potential of recombinant non-purified botulinum neurotoxin serotypes C and D.

    PubMed

    Moreira, Clóvis; da Cunha, Carlos Eduardo Pouey; Moreira, Gustavo Marçal Schmidt Garcia; Mendonça, Marcelo; Salvarani, Felipe Masiero; Moreira, Ângela Nunes; Conceição, Fabricio Rochedo

    2016-08-01

    Botulinum neurotoxin (BoNT) serotypes C and D are responsible for cattle botulism, a fatal paralytic disease that results in great economic losses in livestock production. Vaccination is the main approach to prevent cattle botulism. However, production of commercially available vaccines (toxoids) involves high risk and presents variation of BoNT production between batches. Such limitations can be attenuated by the development of novel nontoxic recombinant vaccines through a simple and reproducible process. The aim of this study was to evaluate the protective potential of recombinant non-purified botulinum neurotoxin serotypes C and D. Bivalent vaccines containing 200 μg rHCC and rHCD each were formulated in three different ways: (1) purified antigens; (2) recombinant Escherichia coli bacterins; (3) recombinant E. coli cell lysates (supernatant and inclusion bodies). Guinea pigs immunized subcutaneously with recombinant formulations developed a protective immune response against the respective BoNTs as determined by a mouse neutralization bioassay with pooled sera. Purified recombinant antigens were capable of inducing 13 IU/mL antitoxin C and 21 IU/mL antitoxin D. Similarly, both the recombinant bacterins and the cell lysate formulations were capable of inducing 12 IU/mL antitoxin C and 20 IU/mL antitoxin D. These values are two times as high as compared to values induced by the commercial toxoid used as control, and two to ten times as high as the minimum amount required by the Brazilian Ministry of Agriculture, Livestock and Food Supply (MAPA), respectively. Therefore, we used a practical, industry-friendly, and efficient vaccine production process that resulted in formulations capable of inducing protective immune response (neutralizing antitoxins) against botulism serotypes C and D.

  3. Phosphorylation of the purified cardiac ryanodine receptor by exogenous and endogenous protein kinases.

    PubMed Central

    Hohenegger, M; Suko, J

    1993-01-01

    The ryanodine receptor is the main Ca(2+)-release structure in skeletal and cardiac sarcoplasmic reticulum. In both tissues, phosphorylation of the ryanodine receptor has been proposed to be involved in the regulation of Ca2+ release. In the present study, we have examined the ability of the purified cardiac ryanodine receptor to serve as a substrate for phosphorylation by exogenously added catalytic subunit of the cyclic AMP (cAMP)-dependent protein kinase (PK-A), cyclic GMP (cGMP)-dependent protein kinase (PK-G), or calmodulin-dependent protein kinase (PK-CaM). A large amount of phosphate incorporation was observed for PK-CaM (938 +/- 48 pmol of Pi/mg of purified channel protein), whereas the level of phosphorylation was considerably lower with PK-A or PK-G (345 +/- 139 and 96 +/- 6 pmol/mg respectively). In addition, endogenous PK-CaM activity co-migrates with the ryanodine receptor through several steps of purification, suggesting a strong association of the two proteins. This endogenous PK-CaM activity is abolished by a PK-CaM-specific synthetic peptide inhibitor. Endogenous cAMP- and cGMP-dependent phosphorylation was not observed in the purified ryanodine-receptor preparation. Taken together, these observations imply that PK-CaM is the physiologically relevant protein kinase, capable of phosphorylating the channel protein to a minimum stoichiometry of 2 mol of Pi per mol of tetramer. Images Figure 2 Figure 3 Figure 4 PMID:8257417

  4. Growth in and breakdown of purified rabbit small intestinal mucin by Yersinia enterocolitica.

    PubMed Central

    Mantle, M; Rombough, C

    1993-01-01

    The mucus lining of the gastrointestinal tract serves as a protective barrier over the epithelial surface that must be crossed by invading bacteria seeking entry into the mucosa. The gel-forming component of mucus is mucin, a large polymeric glycoprotein. The present study examined the growth of Yersinia enterocolitica (with and without its virulence plasmid) in purified rabbit small intestinal mucin and the ability of bacteria to degrade mucin. Both virulent and nonvirulent organisms showed enhanced growth in mucin-supplemented media compared with unsupplemented media, but only at 37 degrees C and not at 25 degrees C. The effects of mucin were not specific because medium supplemented with bovine serum albumin also enhanced bacterial growth at 37 degrees C. Purified mucin was broken down into lower-molecular-weight components (assessed by monitoring its elution profile on a Sepharose CL-2B column) by plasmid-bearing Y. enterocolitica but not by plasmid-cured organisms. Culturing virulent Y. enterocolitica at 25 degrees C completely suppressed its capacity to degrade mucin, suggesting that this activity depends on plasmid expression. These results were confirmed in similar studies with purified rabbit colonic mucin. Mucin-degrading activity could be demonstrated in spent culture media from virulent Y. enterocolitica incubated at 37 degrees C but not in bacterial membrane preparations. Changes in the elution profiles of small intestinal and colonic mucins exposed to plasmid-bearing Y. enterocolitica at 37 degrees C were consistent with proteolytic depolymerization. The ability to grow well in mucin may help Y. enterocolitica to colonize the intestine, while the production of a mucin-degrading enzyme(s) by plasmid-bearing organisms may assist pathogenic strains to solubilize and penetrate the mucus gel layer. PMID:8406802

  5. The respiratory burst oxidase of human neutrophils. Further studies of the purified enzyme.

    PubMed

    Glass, G A; DeLisle, D M; DeTogni, P; Gabig, T G; Magee, B H; Markert, M; Babior, B M

    1986-10-05

    A superoxide-forming oxidase from activated human neutrophil membranes was solubilized by two slightly different methods, then purified by "dye-affinity" chromatography. Kinetic studies of the purified preparations gave Vmax values of 5-10 mumol of O-2/min/mg of protein, and Km values for NADH and NADPH that were in reasonable agreement with values determined previously using particulate and crude solubilized preparations of the respiratory burst oxidase. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed prominent bands at 67, 48, and 32 kDa, together with some minor contaminants, whereas gel electrophoresis under non-denaturing conditions gave a single major band that when eluted and re-electrophoresed in the presence of sodium dodecyl sulfate showed bands at 67, 48, 32 kDa. We believe that all three bands represent oxidase components. The flavin content of the purified enzyme was 20.4 +/- 2.0 S.E. pmol of FAD/microgram of protein, whereas heme averaged 0.1 +/- 0.02 pmol/microgram and ubiquinone could not be detected. Assuming that the enzyme is composed of one 67-kDa subunit, one 48-kDa subunit, and one 32-kDa subunit (i.e. that its molecular mass is approximately 150 kDa), it can be calculated to have a turnover number of 700-1500 min-1, in agreement with a value reported previously for oxidase in a particulate O-2-forming system (Cross, A. R., Parkinson, J. F., and Jones, O. T. G. (1985) Biochem. J. 226, 881-884), and to contain the following quantities of redox carriers (mol/mol): FAD, 3.0; heme, 0.015; ubiquinone, less than 0.06. It remains to be determined whether this preparation represents the complete respiratory burst oxidase or is only the pyridine nucleotide dehydrogenating component of a more complex enzyme.

  6. Substrate Specificity of Purified Recombinant Chicken β-Carotene 9',10'-Oxygenase (BCO2).

    PubMed

    Dela Seña, Carlo; Sun, Jian; Narayanasamy, Sureshbabu; Riedl, Kenneth M; Yuan, Yan; Curley, Robert W; Schwartz, Steven J; Harrison, Earl H

    2016-07-08

    Provitamin A carotenoids are oxidatively cleaved by β-carotene 15,15'-dioxygenase (BCO1) at the central 15-15' double bond to form retinal (vitamin A aldehyde). Another carotenoid oxygenase, β-carotene 9',10'-oxygenase (BCO2) catalyzes the oxidative cleavage of carotenoids at the 9'-10' bond to yield an ionone and an apo-10'-carotenoid. Previously published substrate specificity studies of BCO2 were conducted using crude lysates from bacteria or insect cells expressing recombinant BCO2. Our attempts to obtain active recombinant human BCO2 expressed in Escherichia coli were unsuccessful. We have expressed recombinant chicken BCO2 in the strain E. coli BL21-Gold (DE3) and purified the enzyme by cobalt ion affinity chromatography. Like BCO1, purified recombinant chicken BCO2 catalyzes the oxidative cleavage of the provitamin A carotenoids β-carotene, α-carotene, and β-cryptoxanthin. Its catalytic activity with β-carotene as substrate is at least 10-fold lower than that of BCO1. In further contrast to BCO1, purified recombinant chicken BCO2 also catalyzes the oxidative cleavage of 9-cis-β-carotene and the non-provitamin A carotenoids zeaxanthin and lutein, and is inactive with all-trans-lycopene and β-apocarotenoids. Apo-10'-carotenoids were detected as enzymatic products by HPLC, and the identities were confirmed by LC-MS. Small amounts of 3-hydroxy-β-apo-8'-carotenal were also consistently detected in BCO2-β-cryptoxanthin reaction mixtures. With the exception of this activity with β-cryptoxanthin, BCO2 cleaves specifically at the 9'-10' bond to produce apo-10'-carotenoids. BCO2 has been shown to function in preventing the excessive accumulation of carotenoids, and its broad substrate specificity is consistent with this.

  7. New Method Developed To Purify Single Wall Carbon Nanotubes for Aerospace Applications

    NASA Technical Reports Server (NTRS)

    Lebron, Marisabel; Meador, Michael A.

    2003-01-01

    Single wall carbon nanotubes have attracted considerable attention because of their remarkable mechanical properties and electrical and thermal conductivities. Use of these materials as primary or secondary reinforcements in polymers or ceramics could lead to new materials with significantly enhanced mechanical strength and electrical and thermal conductivity. Use of carbon-nanotube-reinforced materials in aerospace components will enable substantial reductions in component weight and improvements in durability and safety. Potential applications for single wall carbon nanotubes include lightweight components for vehicle structures and propulsion systems, fuel cell components (bipolar plates and electrodes) and battery electrodes, and ultra-lightweight materials for use in solar sails. A major barrier to the successful use of carbon nanotubes in these components is the need for methods to economically produce pure carbon nanotubes in large enough quantities to not only evaluate their suitability for certain applications but also produce actual components. Most carbon nanotube synthesis methods, including the HiPCO (high pressure carbon monoxide) method developed by Smalley and others, employ metal catalysts that remain trapped in the final product. These catalyst impurities can affect nanotube properties and accelerate their decomposition. The development of techniques to remove most, if not all, of these impurities is essential to their successful use in practical applications. A new method has been developed at the NASA Glenn Research Center to purify gram-scale quantities of single wall carbon nanotubes. This method, a modification of a gas phase purification technique previously reported by Smalley and others, uses a combination of high-temperature oxidations and repeated extractions with nitric and hydrochloric acid. This improved procedure significantly reduces the amount of impurities (catalyst and nonnanotube forms of carbon) within the nanotubes, increasing

  8. Toxicity and immunogenicity of purified Haemophilus ducreyi cytolethal distending toxin in a rabbit model.

    PubMed

    Wising, Catharina; Svensson, Liselott A; Ahmed, Hinda J; Sundaeus, Vivianne; Ahlman, Karin; Jonsson, Ing-Marie; Mölne, Lena; Lagergård, Teresa

    2002-08-01

    The cytolethal distending toxin of Haemophilus ducreyi (HdCDT) is a three-component toxin that induces the arrest of the mammalian cell cycle in the G2 phase. All of the individual gene products, CdtA, CdtB and CdtC, are required for toxic activity on cultured mammalian cells. The CdtB component alone exerts nuclease activity. The individual HdCDT components were purified by affinity chromatography or ion-exchange chromatography followed by gel-filtration. HdCDT was reconstituted and purified by the immobilization of a GST-CdtB fusion on a GSTrap column and the subsequent addition of cell sonicates from Escherichia coli recombinants that produced CdtA and CdtC. The purified HdCDT preparation contained all three CDT proteins, as detected by immuno-blotting, and had high cytotoxic activity (10(6)CPU/ml). Immunization of rabbits with the HdCDT complex and with the individual CdtA, CdtB and CdtC proteins elicited high titres of antibodies, as detected by ELISA. All of the immune sera had toxin-neutralizing activities. The pathological effects of the HdCDT complex were investigated in rabbits, since the proliferation of two rabbit cell lines, SIRC and RK-13, was inhibited by HdCDT. Intradermal injection of HdCDT (1, 10, 50 and 100microg protein) into naive rabbits resulted in dose-dependent skin reactions (erythema) about 24h after injection. Similar effects were not observed when the individual HdCDT proteins were injected. HdCDT injection into immune rabbits resulted in dose-dependent skin responses that were characterized by both erythema and oedema. Histological evaluation of the 24-h lesions in naive rabbits that were injected with HdCDT, revealed moderate levels of inflammatory cells, which were mainly granulocytes and macrophages, and dilatation of blood vessels. The skin reactions in HdCDT-injected immunized rabbits showed pronounced vascular changes and extensive infiltration of inflammatory cells, including eosinophils. All of the pathological changes healed

  9. Tyrosine-specific phosphorylation of calmodulin by the insulin receptor kinase purified from human placenta.

    PubMed Central

    Sacks, D B; Fujita-Yamaguchi, Y; Gale, R D; McDonald, J M

    1989-01-01

    It has previously been demonstrated that calmodulin can be phosphorylated in vitro and in vivo by both tyrosine-specific and serine/threonine protein kinase. We demonstrate here that the insulin receptor tyrosine kinase purified from human placenta phosphorylates calmodulin. The highly purified receptors (prepared by insulin-Sepharose chromatography) were 5-10 times more effective in catalysing the phosphorylation of calmodulin than an equal number of partially purified receptors (prepared by wheat-germ agglutinin-Sepharose chromatography). Phosphorylation occurred exclusively on tyrosine residues, up to a maximum of 1 mol [0.90 +/- 0.14 (n = 5)] of phosphate incorporated/mol of calmodulin. Phosphorylation of calmodulin was dependent on the presence of certain basic proteins and divalent cations. Some of these basic proteins, i.e. polylysine, polyarginine, polyornithine, protamine sulphate and histones H1 and H2B, were also able to stimulate the phosphorylation of calmodulin via an insulin-independent activation of the receptor tyrosine kinase. Addition of insulin further increased incorporation of 32P into calmodulin. The magnitude of the effect of insulin was dependent on the concentration and type of basic protein used, ranging from 0.5- to 9.0-fold stimulation. Maximal phosphorylation of calmodulin was obtained at an insulin concentration of 10(-10) M, with half-maximal effect at 10(-11) M. Either Mg2+ or Mn2+ was necessary to obtain phosphorylation, but Mg2+ was far more effective than Mn2+. In contrast, maximal phosphorylation of calmodulin was observed in the absence of Ca2+. Inhibition of phosphorylation was observed as free Ca2+ concentration exceeded 0.1 microM, with almost complete inhibition at 30 microM free Ca2+. The Km for calmodulin was approx. 0.1 microM. To gain further insight into the effects of basic proteins in this system, we examined the binding of calmodulin to the insulin receptor and the polylysine. Calmodulin binds to the insulin

  10. Interaction of purified alternative oxidase from thermogenic Arum maculatum with pyruvate.

    PubMed

    Carré, J E; Affourtit, C; Moore, A L

    2011-01-21

    Plant alternative oxidase (AOX) activity in isolated mitochondria is regulated by carboxylic acids, but reaction and regulatory mechanisms remain unclear. We show that activity of AOX protein purified from thermogenic Arum maculatum spadices is sensitive to pyruvate and glyoxylate but not succinate. Rapid, irreversible AOX inactivation occurs in the absence of pyruvate, whether or not duroquinol oxidation has been initiated, and is insensitive to duroquinone. Our data indicate that pyruvate stabilises an active conformation of AOX, increasing the population of active protein in a manner independent of reducing substrate and product, and are thus consistent with an exclusive effect of pyruvate on the enzyme's apparent V(max).

  11. Studies on the interactions between purified bovine caseins and alkaline-earth-metalions

    PubMed Central

    Dickson, I. R.; Perkins, D. J.

    1971-01-01

    1. Alkaline-earth-metal cations at low concentrations form soluble complexes with bovine caseins. The relative order of binding capacities is: Mg2+>Ca2+>Ba2+>Sr2+. 2. The cations interact with both free ionized carboxyl groups of aspartic acid and glutamic acid and with monoester phosphate groups covalently bound to serine and threonine; at low concentrations of the cations interactions are predominantly with the phosphate groups. 3. The order of binding capacities for purified components of the casein complex is: αs1-casein>β-casein>κ-casein. PMID:5166590

  12. Purified Lesser weever fish venom (Trachinus vipera) induces eryptosis, apoptosis and cell cycle arrest

    PubMed Central

    Fezai, Myriam; Slaymi, Chaker; Ben-Attia, Mossadok; Lang, Florian; Jemaà, Mohamed

    2016-01-01

    Accidents caused by the sting of Trachinus vipera (known as Lesser weever fish) are relatively common in shallow waters of the Mediterranean. Symptoms after the sting vary from severe pain to edema or even tissue necrosis in some cases. Here we show that purified Lesser weever fish venom induces eryptosis, the suicidal erythrocyte death, and apoptosis of human colon carcinoma cells. The venom leads to erythrocyte shrinkage, phosphatidylserine translocation and increased intracellular Ca2+, events typical for eryptosis. According to mitochondrial staining cancer cells dyed after the activation of the intrinsic apoptotic pathway. Trachinus vipera venom further causes cell cycle arrest. PMID:27995979

  13. Hydrodynamic and Membrane Binding Properties of Purified Rous Sarcoma Virus Gag Protein

    SciTech Connect

    Dick, Robert A.; Datta, Siddhartha A. K.; Nanda, Hirsh; Fang, Xianyang; Wen, Yi; Barros, Marilia; Wang, Yun-Xing; Rein, Alan; Vogt, Volker M.; Sundquist, W. I.

    2016-05-06

    Previously, no retroviral Gag protein has been highly purified in milligram quantities and in a biologically relevant and active form. We have purified Rous sarcoma virus (RSV) Gag protein and in parallel several truncation mutants of Gag and have studied their biophysical properties and membrane interactionsin vitro. RSV Gag is unusual in that it is not naturally myristoylated. From its ability to assemble into virus-like particlesin vitro, we infer that RSV Gag is biologically active. By size exclusion chromatography and small-angle X-ray scattering, Gag in solution appears extended and flexible, in contrast to previous reports on unmyristoylated HIV-1 Gag, which is compact. However, by neutron reflectometry measurements of RSV Gag bound to a supported bilayer, the protein appears to adopt a more compact, folded-over conformation. At physiological ionic strength, purified Gag binds strongly to liposomes containing acidic lipids. This interaction is stimulated by physiological levels of phosphatidylinositol-(4,5)-bisphosphate [PI(4,5)P2] and by cholesterol. However, unlike HIV-1 Gag, RSV Gag shows no sensitivity to acyl chain saturation. In contrast with full-length RSV Gag, the purified MA domain of Gag binds to liposomes only weakly. Similarly, both an N-terminally truncated version of Gag that is missing the MA domain and a C-terminally truncated version that is missing the NC domain bind only weakly. These results imply that NC contributes to membrane interactionin vitro, either by directly contacting acidic lipids or by promoting Gag multimerization.

    Retroviruses like HIV assemble at and bud from the plasma membrane of cells. Assembly requires the interaction between thousands of Gag molecules to form a lattice. Previous work indicated that lattice formation at the plasma membrane is influenced by the conformation of monomeric HIV. We have extended this work to the more tractable RSV Gag. Our

  14. Specific IgE response to purified and recombinant allergens in latex allergy

    PubMed Central

    Kurup, Viswanath P; Sussman, Gordon L; Yeang, Hoong Y; Elms, Nancy; Breiteneder, Heimo; Arif, Siti AM; Kelly, Kevin J; Bansal, Naveen K; Fink, Jordan N

    2005-01-01

    Background In recent years, allergy to natural rubber latex has emerged as a major allergy among certain occupational groups and patients with underlying diseases. The sensitization and development of latex allergy has been attributed to exposure to products containing residual latex proteins. Although improved manufacturing procedures resulted in a considerable reduction of new cases, the potential risk for some patient groups is still great. In addition the prevalent cross-reactivity of latex proteins with other food allergens poses a major concern. A number of purified allergens and a few commercial kits are currently available, but no concerted effort was undertaken to evaluate them. Methods We studied 11 purified latex allergens, Hev b 1 to Hev b 10, and Hev b 13 along with several crude allergen extracts and two commercial ImmunoCAP assays to evaluate specific IgE antibody in the sera from latex allergic patients and controls. Health care workers and spina bifida patients with clinical symptoms of latex allergy, spina bifida patients without latex allergy, and non-atopic health care workers have been studied. Results The results suggest that Hev b 2, 5, 6, and 13 together identified over 80 percent health care workers with latex allergy, while Hev b 6 along with Hev b 1 or 3 detected specific IgE antibody in all sera studied from patients with spina bifida and latex allergy. The ImmunoCAP results using both Hev b 5 amplified and non-amplified closely agreed with the clinical diagnosis of latex allergy in health care workers and in spina bifida. Conclusion Although the purified allergens and crude extracts reacted diversely with IgE from different patient groups, the results indicated that use of certain combinations of purified recombinant antigens will be useful in commercial kits or in in-house assays for detecting specific IgE antibody in the sera. The results suggest that a combination of Hev b 2, 3, 5, 6, and 13 together detected specific IgE in 80% of

  15. Applying the INTACT method to purify endosperm nuclei and to generate parental-specific epigenome profiles.

    PubMed

    Moreno-Romero, Jordi; Santos-González, Juan; Hennig, Lars; Köhler, Claudia

    2017-02-01

    The early endosperm tissue of dicot species is very difficult to isolate by manual dissection. This protocol details how to apply the INTACT (isolation of nuclei tagged in specific cell types) system for isolating early endosperm nuclei of Arabidopsis at high purity and how to generate parental-specific epigenome profiles. As a Protocol Extension, this article describes an adaptation of an existing Nature Protocol that details the use of the INTACT method for purification of root nuclei. We address how to obtain the INTACT lines, generate the starting material and purify the nuclei. We describe a method that allows purity assessment, which has not been previously addressed. The purified nuclei can be used for ChIP and DNA bisulfite treatment followed by next-generation sequencing (seq) to study histone modifications and DNA methylation profiles, respectively. By using two different Arabidopsis accessions as parents that differ by a large number of single-nucleotide polymorphisms (SNPs), we were able to distinguish the parental origin of epigenetic modifications. Our protocol describes the only working method to our knowledge for generating parental-specific epigenome profiles of the early Arabidopsis endosperm. The complete protocol, from silique collection to finished libraries, can be completed in 2 d for bisulfite-seq (BS-seq) and 3 to 4 d for ChIP-seq experiments.This protocol is an extension to: Nat. Protoc. 6, 56-68 (2011); doi:10.1038/nprot.2010.175; published online 16 December 2010.

  16. Decolorization of the textile dyes using purified banana pulp polyphenol oxidase.

    PubMed

    Jadhav, Umesh U; Dawkar, Vishal V; Jadhav, Mital U; Govindwar, Sanjay P

    2011-04-01

    Polyphenol oxidase (PPO) purified using DEAE-cellulose and Biogel P-100 column chromatography from banana pulp showed 12.72-fold activity and 2.49% yield. The optimum temperature and pH were found to be 30 degrees C and 7.0, respectively for its activity. Catechol was found to be a suitable substrate for banana pulp PPO that showed V(max), 0.041 mM min(-1) and K(m), 1.6 mM. The enzyme activity was inhibited by sodium metabisulfite, citric acid, cysteine, and beta-mercaptoethanol at 10 mM concentration. The purified enzyme could decolorize (90%) Direct Red 5B (160 microg mL(-1)) dye within 48 h and Direct Blue GLL (400 microg mL(-1)) dye up to 85% within 90 h. The GC-MS analysis indicated the presence of 4-hydroxy-benzenesulfonic acid and Naphthalene-1,2,3,6-tetraol in the degradation products of Direct Red 5B, and 5-(4-Diazenyl-naphthalene-1-ylazo)-8-hydroxy-naphthalene-2-sulfonic acid and 2-(4-Diazenyl-naphthalene-1-ylazo)-benzenesulfonic acid in the degradation products of Direct Blue GLL.

  17. Proteome Profile of Starch Granules Purified from Rice (Oryza sativa) Endosperm

    PubMed Central

    Xing, Shihai; Meng, Xiaoxi; Zhou, Lihui; Mujahid, Hana; Zhao, Chunfang; Zhang, Yadong; Wang, Cailin; Peng, Zhaohua

    2016-01-01

    Starch is the most important food energy source in cereals. Many of the known enzymes involved in starch biosynthesis are partially or entirely granule-associated in the endosperm. Studying the proteome of rice starch granules is critical for us to further understand the mechanisms underlying starch biosynthesis and packaging of starch granules in rice amyloplasts, consequently for the improvement of rice grain quality. In this article, we developed a protocol to purify starch granules from mature rice endosperm and verified the quality of purified starch granules by microscopy observations, I2 staining, and Western blot analyses. In addition, we found the phenol extraction method was superior to Tris-HCl buffer extraction method with respect to the efficiency in recovery of starch granule associated proteins. LC-MS/MS analysis showed identification of already known starch granule associated proteins with high confidence. Several proteins reported to be involved in starch synthesis in prior genetic studies in plants were also shown to be enriched with starch granules, either directly or indirectly, in our studies. In addition, our results suggested that a few additional candidate proteins may also be involved in starch synthesis. Furthermore, our results indicated that some starch synthesis pathway proteins are subject to protein acetylation modification. GO analysis and KEGG pathway enrichment analysis showed that the identified proteins were mainly located in plastids and involved in carbohydrate metabolism. This study substantially advances the understanding of the starch granule associated proteome in rice and post translational regulation of some starch granule associated proteins. PMID:27992503

  18. In vivo behavior of detergent-solubilized purified rabbit thrombomodulin on intravenous injection into rabbits

    SciTech Connect

    Ehrlich, H.J.; Esmon, N.L.; Bang, N.U. )

    1990-02-01

    Thrombomodulin is a thrombin endothelial cell membrane receptor. The thrombomodulin-thrombin complex rapidly activates protein C resulting in anticoagulant activity. We investigated the anticoagulant effects and pharmacokinetic behavior of detergent-solubilized purified rabbit thrombomodulin labeled with iodine 125 when intravenously injected into rabbits. Thrombomodulin half-life (t1/2) was determined by tracking the 125I-radiolabeled protein and the biologic activity as determined by the prolongation of the activated partial thromboplastin time (APTT) and thrombin clotting time (TCT). When 200 micrograms/kg 125I-thrombomodulin was injected into rabbits, the APTT and TCT were immediately prolonged, whereas no effect on the prothrombin time was seen. In vitro calibration curves enabled us to convert the prolongations of the clotting times into micrograms per milliliter thrombomodulin equivalents. The best fit (r greater than 0.99) for the disappearance curves was provided by a two-compartment model with mean t1/2 alpha (distribution phase) of 18 minutes for 125I, 12 minutes for APTT, and 20 minutes for TCT, and mean t1/2 beta (elimination phase) of 385 minutes for 125I, 460 for APTT, and 179 for TCT. The administration of two doses of endotoxin (50 micrograms/kg) 24 hours apart did not accelerate the turnover rate of 125I-thrombomodulin as measured by the disappearance of 125I from the circulation. Thus, detergent-solubilized purified thrombomodulin administered intravenously circulates in a biologically active form for appreciable time periods.

  19. Application of decolourized and partially purified polygalacturonase and α-amylase in apple juice clarification

    PubMed Central

    Dey, Tapati Bhanja; Banerjee, Rintu

    2014-01-01

    Polygalacturonase and α-amylase play vital role in fruit juice industry. In the present study, polygalacturonase was produced by Aspergillus awamori Nakazawa MTCC 6652 utilizing apple pomace and mosambi orange (Citrus sinensis var mosambi) peels as solid substrate whereas, α-amylase was produced from A. oryzae (IFO-30103) using wheat bran by solid state fermentation (SSF) process. These carbohydrases were decolourized and purified 8.6-fold, 34.8-fold and 3.5-fold, respectively by activated charcoal powder in a single step with 65.1%, 69.8% and 60% recoveries, respectively. Apple juice was clarified by these decolourized and partially purified enzymes. In presence of 1% polygalacturonase from mosambi peels (9.87 U/mL) and 0.4% α-amylase (899 U/mL), maximum clarity (%T660nm = 97.0%) of juice was attained after 2 h of incubation at 50 °C in presence of 10 mM CaCl2. Total phenolic content of juice was reduced by 19.8% after clarification, yet with slightly higher %DPPH radical scavenging property. PMID:24948919

  20. Replication of adenovirus type 4 DNA by a purified fraction from infected cells.

    PubMed Central

    Temperley, S M; Hay, R T

    1991-01-01

    An extract from Adenovirus type 4 infected HeLa cells was fractionated by ion-exchange and DNA affinity chromatography. One fraction, which bound tightly to single stranded DNA, contained predominantly a protein of apparent molecular weight 65,000 and three less abundant proteins. Immunological cross-reactivity with adenovirus type 2 proteins confirmed the presence of preterminal protein and indicated that the abundant species was the virus coded DNA binding protein. This fraction contained an aphidicolin resistant DNA polymerase activity and in the presence of a linearised plasmid containing the adenovirus type 4 origin of DNA replication efficient transfer of dCMP onto preterminal protein, indicative of initiation, was observed. Furthermore, addition of all four deoxyribonucleotide triphosphates and an ATP regenerating system resulted in the elongation of initiated molecules to generate plasmid molecules covalently attached to preterminal protein. Adenovirus type 4 DNA binding protein was extensively purified from crude adenovirus-4 infected HeLa extract by immunoaffinity chromatography using a monoclonal antibody raised against adenovirus type 2 DNA binding protein. A low level of initiation of DNA replication was detected in the fraction depleted of DNA binding protein but activity was restored by addition of purified DNA binding protein. DNA binding protein therefore plays an important role in the initiation of Ad4 DNA replication. Images PMID:1829516

  1. Chitosanase purified from bacterial isolate Bacillus licheniformis of ruined vegetables displays broad spectrum biofilm inhibition.

    PubMed

    Muslim, Sahira Nsayef; Al-Kadmy, Israa M S; Hussein, Nadheema Hammood; Mohammed Ali, Alaa Naseer; Taha, Buthainah Mohammed; Aziz, Sarah Naji; Kheraif, Abdulaziz Abdullah Al; Divakar, Darshan Devang; Ramakrishnaiah, Ravikumar

    2016-11-01

    A number of bacterial species produces chitosanases which has variety of applications because of its high biodegradability, non-toxicity and antimicrobial assets. In the present study chitosanase is purified from new bacterial species Bacillus licheniformis from spoiled vegetable. This novel strain of Bacillus licheniformis isolated from spoilt cucumber and pepper samples has the ability to produce the chitosanase enzyme when grown on chitosan substrate. Study also examined its antibiofilm properties against diverse bacterial species with biofilm forming ability. The purified chitosanase inhibited the biofilm formation ability for all Gram-negative and Gram-positive biofilm-forming bacteria [biofilm producers] tested in this study in congo red agar and microtiter plate's methods. Highly antibiofilm activity of chitosanase was recorded against Pseudomonas aeruginosa followed by Klebsiella pneumoniae with reduction of biofilm formation upto 22 and 29%, respectively compared with [100] % of control. Biofilm formation has multiple role including ability to enhance resistance and self-protection from external stress. This chitosanase has promising benefit as antibiofilm agent against biofilm forming pathogenic bacteria and has promising application as alternative antibiofilm agents to combat the growing number of multidrug resistant pathogen-associated infections, especially in situation where biofilms are involved.

  2. A method of batch-purifying microalgae with multiple antibiotics at extremely high concentrations

    NASA Astrophysics Data System (ADS)

    Han, Jichang; Wang, Song; Zhang, Lin; Yang, Guanpin; Zhao, Lu; Pan, Kehou

    2016-01-01

    Axenic microalgal strains are highly valued in diverse microalgal studies and applications. Antibiotics, alone or in combination, are often used to avoid bacterial contamination during microalgal isolation and culture. In our preliminary trials, we found that many microalgae ceased growing in antibiotics at extremely high concentrations but could resume growth quickly when returned to an antibiotics-free liquid medium and formed colonies when spread on a solid medium. We developed a simple and highly efficient method of obtaining axenic microalgal cultures based on this observation. First, microalgal strains of different species or strains were treated with a mixture of ampicillin, gentamycin sulfate, kanamycin, neomycin and streptomycin (each at a concentration of 600 mg/L) for 3 days; they were then transferred to antibiotics-free medium for 5 days; and finally they were spread on solid f/2 media to allow algal colonies to form. With this method, five strains of Nannochloropsis sp. (Eustigmatophyceae), two strains of Cylindrotheca sp. (Bacillariophyceae), two strains of Tetraselmis sp. (Chlorodendrophyceae) and one strain of Amphikrikos sp. (Trebouxiophyceae) were purified successfully. The method shows promise for batch-purifying microalgal cultures.

  3. Comparison of solubilized and purified plasma membrane and nuclear insulin receptors

    SciTech Connect

    Wong, K.Y.; Hawley, D.; Vigneri, R.; Goldfine, I.D.

    1988-01-12

    Prior studies have detected biochemical and immunological differences between insulin receptors in plasma membranes and isolated nuclei. To further investigate these receptors, they were solubilized in Triton X-100 partially purified by wheat germ agglutinin-agarose chromatography. In these preparations, the nuclear and plasma membrane receptors had very similar pH optima (pH 8.0) and reactivities to a group of polyclonal antireceptor antibodies. Further, both membrane preparations had identical binding activities when labeled insulin was competed for by unlabeled insulin (50% inhibition at 800 pM). Next, nuclear and plasma membranes were solubilized and purified to homogeneity by wheat germ agglutinin-agarose and insulin-agarose chromatography. In both receptors, labeled insulin was covalently cross-linked to a protein of 130 kilodaltons representing the insulin receptor ..cap alpha.. subunit. When preparations of both receptors were incubated with insulin and then adenosine 5'-(..gamma..-/sup 32/P)triphosphate, a protein of 95 kilodaltons representing the insulin receptor ..beta.. subunit was phosphorylated in a dose-dependent manner. These studies indicate, therefore, that solubilized plasma membrane and nuclear insulin receptors have similar structures and biochemical properties, and they suggest that they are the same (or very similar) proteins.

  4. Characterization of Affinity-Purified Isoforms of Acinetobacter calcoaceticus Y1 Glutathione Transferases

    PubMed Central

    Chee, Chin-Soon; Tan, Irene Kit-Ping; Alias, Zazali

    2014-01-01

    Glutathione transferases (GST) were purified from locally isolated bacteria, Acinetobacter calcoaceticus Y1, by glutathione-affinity chromatography and anion exchange, and their substrate specificities were investigated. SDS-polyacrylamide gel electrophoresis revealed that the purified GST resolved into a single band with a molecular weight (MW) of 23 kDa. 2-dimensional (2-D) gel electrophoresis showed the presence of two isoforms, GST1 (pI 4.5) and GST2 (pI 6.2) with identical MW. GST1 was reactive towards ethacrynic acid, hydrogen peroxide, 1-chloro-2,4-dinitrobenzene, and trans,trans-hepta-2,4-dienal while GST2 was active towards all substrates except hydrogen peroxide. This demonstrated that GST1 possessed peroxidase activity which was absent in GST2. This study also showed that only GST2 was able to conjugate GSH to isoproturon, a herbicide. GST1 and GST2 were suggested to be similar to F0KLY9 (putative glutathione S-transferase) and F0KKB0 (glutathione S-transferase III) of Acinetobacter calcoaceticus strain PHEA-2, respectively. PMID:24892084

  5. Continuous Quadrupole Magnetic Separation of Islets during Digestion Improves Purified Porcine Islet Viability

    PubMed Central

    Kumar Sajja, Venkata Sunil; Rizzari, Michael D.; Scott III, William E.; Kitzmann, Jennifer P.; Kennedy, David J.; Todd, Paul W.; Balamurugan, Appakalai N.; Hering, Bernhard J.

    2016-01-01

    Islet transplantation (ITx) is an emerging and promising therapy for patients with uncontrolled type 1 diabetes. The islet isolation and purification processes require exposure to extended cold ischemia, warm-enzymatic digestion, mechanical agitation, and use of damaging chemicals for density gradient separation (DG), all of which reduce viable islet yield. In this paper, we describe initial proof-of-concept studies exploring quadrupole magnetic separation (QMS) of islets as an alternative to DG to reduce exposure to these harsh conditions. Three porcine pancreata were split into two parts, the splenic lobe (SPL) and the combined connecting/duodenal lobes (CDL), for paired digestions and purifications. Islets in the SPL were preferentially labeled using magnetic microparticles (MMPs) that lodge within the islet microvasculature when infused into the pancreas and were continuously separated from the exocrine tissue by QMS during the collection phase of the digestion process. Unlabeled islets from the CDL were purified by conventional DG. Islets purified by QMS exhibited significantly improved viability (measured by oxygen consumption rate per DNA, p < 0.03) and better morphology relative to control islets. Islet purification by QMS can reduce the detrimental effects of prolonged exposure to toxic enzymes and density gradient solutions and substantially improve islet viability after isolation. PMID:27843954

  6. Continuous Quadrupole Magnetic Separation of Islets during Digestion Improves Purified Porcine Islet Viability.

    PubMed

    Weegman, Bradley P; Kumar Sajja, Venkata Sunil; Suszynski, Thomas M; Rizzari, Michael D; Scott Iii, William E; Kitzmann, Jennifer P; Mueller, Kate R; Hanley, Thomas R; Kennedy, David J; Todd, Paul W; Balamurugan, Appakalai N; Hering, Bernhard J; Papas, Klearchos K

    2016-01-01

    Islet transplantation (ITx) is an emerging and promising therapy for patients with uncontrolled type 1 diabetes. The islet isolation and purification processes require exposure to extended cold ischemia, warm-enzymatic digestion, mechanical agitation, and use of damaging chemicals for density gradient separation (DG), all of which reduce viable islet yield. In this paper, we describe initial proof-of-concept studies exploring quadrupole magnetic separation (QMS) of islets as an alternative to DG to reduce exposure to these harsh conditions. Three porcine pancreata were split into two parts, the splenic lobe (SPL) and the combined connecting/duodenal lobes (CDL), for paired digestions and purifications. Islets in the SPL were preferentially labeled using magnetic microparticles (MMPs) that lodge within the islet microvasculature when infused into the pancreas and were continuously separated from the exocrine tissue by QMS during the collection phase of the digestion process. Unlabeled islets from the CDL were purified by conventional DG. Islets purified by QMS exhibited significantly improved viability (measured by oxygen consumption rate per DNA, p < 0.03) and better morphology relative to control islets. Islet purification by QMS can reduce the detrimental effects of prolonged exposure to toxic enzymes and density gradient solutions and substantially improve islet viability after isolation.

  7. The purified mechanosensitive channel TREK-1 is directly sensitive to membrane tension.

    PubMed

    Berrier, Catherine; Pozza, Alexandre; de Lacroix de Lavalette, Agnes; Chardonnet, Solenne; Mesneau, Agnes; Jaxel, Christine; le Maire, Marc; Ghazi, Alexandre

    2013-09-20

    Mechanosensitive channels are detected in all cells and are speculated to play a key role in many functions including osmoregulation, growth, hearing, balance, and touch. In prokaryotic cells, a direct gating of mechanosensitive channels by membrane tension was clearly demonstrated because the purified channels could be functionally reconstituted in a lipid bilayer. No such evidence has been presented yet in the case of mechanosensitive channels from animal cells. TREK-1, a two-pore domain K(+) channel, was the first animal mechanosensitive channel identified at the molecular level. It is the target of a large variety of agents such as volatile anesthetics, neuroprotective agents, and antidepressants. We have produced the mouse TREK-1 in yeast, purified it, and reconstituted the protein in giant liposomes amenable to patch clamp recording. The protein exhibited the expected electrophysiological properties in terms of kinetics, selectivity, and pharmacology. Negative pressure (suction) applied through the pipette had no effect on the channel, but positive pressure could completely and reversibly close the channel. Our interpretation of these data is that the intrinsic tension in the lipid bilayer is sufficient to maximally activate the channel, which can be closed upon modification of the tension. These results indicate that TREK-1 is directly sensitive to membrane tension.

  8. Gas-phase photolytic production of hydroxyl radicals in an ultraviolet purifier for air and surfaces.

    PubMed

    Crosley, David R; Araps, Connie J; Doyle-Eisele, Melanie; McDonald, Jacob D

    2017-02-01

    We have measured the concentration of hydroxyl radicals (OH) produced in the gas phase by a commercially available purifier for air and surfaces, using the time rate of decay of n-heptane added to an environmental chamber. The hydroxyl generator, an Odorox® BOSS™ model, produces the OH through 185-nm photolysis of ambient water vapor. The steady-state concentration of OH produced in the 120 m(3) chamber is, with 2σ error bars, (3.25 ± 0.80) × 10(6) cm(-3). The properties of the hydroxyl generator, in particular the output of the ultraviolet lamps and the air throughput, together with an estimation of the water concentration, were used to predict the amount of OH produced by the device, with no fitted parameters. To relate this calculation to a steady-state concentration, we must estimate the OH loss rate within the chamber owing to reaction with the n-heptane and the 7 ppb of background hydrocarbons that are present. The result is a predicted steady-state concentration in excellent agreement with the measured value. This shows we understand well the processes occurring in the gas phase during operation of this hydroxyl radical purifier.

  9. A purified extract from prickly pear cactus (Opuntia fuliginosa) controls experimentally induced diabetes in rats.

    PubMed

    Trejo-González, A; Gabriel-Ortiz, G; Puebla-Pérez, A M; Huízar-Contreras, M D; Munguía-Mazariegos, M R; Mejía-Arreguín, S; Calva, E

    1996-12-01

    The hypoglycemic activity of a purified extract from prickly pear cactus (Opuntia fuliginosa) was evaluated on STZ-induced diabetic rats. Blood glucose and glycated hemoglobin levels were reduced to normal values by a combined treatment of insulin and Opuntia extract. When insulin was withdrawn from the combined treatment, the prickly pear extract alone maintained normoglycemic state in the diabetic rats. The blood glucose response to administered glucose also showed that the rats receiving the combination treatment of insulin and Opuntia extract for 7 weeks followed by Opuntia extract alone were capable of rapidly returning blood glucose to the levels of the nondiabetic rats. Although the mechanism of action is unknown, the magnitude of the glucose control by the small amount of Opuntia extract required (1 mg/kg body weight per day) preclude a predominant role for dietary fiber. These very encouraging results for diabetes control by the purified extract of this Opuntia cactus make the need for clinical studies in humans evident.

  10. STUDIES ON MEDIATOR PRODUCTION BY HIGHLY PURIFIED HUMAN T AND B LYMPHOCYTES

    PubMed Central

    Rocklin, Ross E.; MacDermott, Richard P.; Chess, Leonard; Schlossman, Stuart F.; David, John R.

    1974-01-01

    Highly purified populations of T and B lymphocytes obtained by affinity column separation were stimulated by antigen and their ability to produce two mediators, migration inhibitory factor (MIF) and lymphocyte mitogenic factor (LMF) was assessed. Both T- and B-cell populations made MIF; the production of MIF was antigen-specific using purified protein derivative of tuberculin, streptokinase-streptodornase, and Candida antigens. The MIF activity from both populations could not be attributed to antigen-antibody complexes as the inhibitory activity eluted from Sephadex G-100 columns in the same region corresponding to mol wt 23,000 daltons. Further studies indicate that the T cells producing MIF are proliferating cells whereas the B cells producing this mediator are not. In contrast, LMF was made only by T cells and not B cells when these populations were stimulated by antigen. The LMF induced the [3H]thymidine incorporation into both T and B cells obtained from donors lacking sensitivity to the antigens used to elicit the factor. Chromatographic studies indicate that LMF eluted from Sephadex G-100 in a fraction of mol wt 23,000 daltons where MIF is also found; however, since B cells produce MIF but not LMF, these two factors appear to be distinct from one another. Some of the implications of these findings are discussed. The explanation for the production or lack of production of MIF by lymphocytes obtained from patients with immunodeficiency disorders requires reinterpretation. PMID:4608321

  11. Lipid Droplets Purified from Drosophila Embryos as an Endogenous Handle for Precise Motor Transport Measurements

    PubMed Central

    Bartsch, Tobias F.; Longoria, Rafael A.; Florin, Ernst-Ludwig; Shubeita, George T.

    2013-01-01

    Molecular motor proteins are responsible for long-range transport of vesicles and organelles. Recent works have elucidated the richness of the transport complex, with multiple teams of similar and dissimilar motors and their cofactors attached to individual cargoes. The interaction among these different proteins, and with the microtubules along which they translocate, results in the intricate patterns of cargo transport observed in cells. High-precision and high-bandwidth measurements are required to capture the dynamics of these interactions, yet the crowdedness in the cell necessitates performing such measurements in vitro. Here, we show that endogenous cargoes, lipid droplets purified from Drosophila embryos, can be used to perform high-precision and high-bandwidth optical trapping experiments to study motor regulation in vitro. Purified droplets have constituents of the endogenous transport complex attached to them and exhibit long-range motility. A novel method to determine the quality of the droplets for high-resolution measurements in an optical trap showed that they compare well with plastic beads in terms of roundness, homogeneity, position sensitivity, and trapping stiffness. Using high-resolution and high-bandwidth position measurements, we demonstrate that we can follow the series of binding and unbinding events that lead to the onset of active transport. PMID:24010661

  12. Daphnia magna negatively affected by chronic exposure to purified Cry-toxins.

    PubMed

    Bøhn, Thomas; Rover, Carina Macagnan; Semenchuk, Philipp Robert

    2016-05-01

    Cry-toxin genes originating from Bacillus thuringiensis are inserted into genetically modified (GM) plants, often called Bt-plants, to provide insect resistance to pests. Significant amounts of Bt-plant residues, and thus Cry-toxins, will be shed to soil and aquatic environments. We exposed Daphnia magna to purified Cry1Ab and Cry2Aa toxins for the full life-span of the animals. We used single toxins in different doses and combinations of toxins and Roundup(®), another potential stressor on the rise in agricultural ecosystems. Animals exposed to 4.5 mg/L (ppm) of Cry1Ab, Cry2Aa and the combination of both showed markedly higher mortality, smaller body size and very low juvenile production compared to controls. Animals exposed to 0.75 mg/L also showed a tendency towards increased mortality but with increased early fecundity compared to the controls. Roundup(®) stimulated animals to strong early reproductive output at the cost of later rapid mortality. We conclude that i) purified Cry-toxins in high concentrations are toxic to D. magna, indicating alternative modes-of-action for these Cry-toxins; ii) Cry-toxins act in combination, indicating that 'stacked events' may have stronger effects on non-target organisms; iii) further studies need to be done on combinatorial effects of multiple Cry-toxins and herbicides that co-occur in the environment.

  13. Purifying Selection Maintains Dosage-Sensitive Genes during Degeneration of the Threespine Stickleback Y Chromosome

    PubMed Central

    White, Michael A.; Kitano, Jun; Peichel, Catherine L.

    2015-01-01

    Sex chromosomes are subject to unique evolutionary forces that cause suppression of recombination, leading to sequence degeneration and the formation of heteromorphic chromosome pairs (i.e., XY or ZW). Although progress has been made in characterizing the outcomes of these evolutionary processes on vertebrate sex chromosomes, it is still unclear how recombination suppression and sequence divergence typically occur and how gene dosage imbalances are resolved in the heterogametic sex. The threespine stickleback fish (Gasterosteus aculeatus) is a powerful model system to explore vertebrate sex chromosome evolution, as it possesses an XY sex chromosome pair at relatively early stages of differentiation. Using a combination of whole-genome and transcriptome sequencing, we characterized sequence evolution and gene expression across the sex chromosomes. We uncovered two distinct evolutionary strata that correspond with known structural rearrangements on the Y chromosome. In the oldest stratum, only a handful of genes remain, and these genes are under strong purifying selection. By comparing sex-linked gene expression with expression of autosomal orthologs in an outgroup, we show that dosage compensation has not evolved in threespine sticklebacks through upregulation of the X chromosome in males. Instead, in the oldest stratum, the genes that still possess a Y chromosome allele are enriched for genes predicted to be dosage sensitive in mammals and yeast. Our results suggest that dosage imbalances may have been avoided at haploinsufficient genes by retaining function of the Y chromosome allele through strong purifying selection. PMID:25818858

  14. Theoretical and experimental investigations on the structures of purified clay and acid-activated clay

    NASA Astrophysics Data System (ADS)

    Yang, Tao; Wen, Xiao-Dong; Li, Junfen; Yang, Liming

    2006-07-01

    The purified and acidified montmorillonite clay were characterized by XRD, BET and TPD. These results show that acidified clay is provided with more surface area and acid sites. For NH 3-TPD, molecular NH 3 desorption on purified clay and acidified clay occurs at temperatures with 873 and 1000 K, respectively. It is shown for the existence for strong acid sites. By two reactions of the tetrahydropyranylation of n-propanol and the esterification of cyclo-2-pentene with acetic acid, it is shown that the acidified clay displays better catalytic activity for above two organic reactions. By density-functional theory (DFT) method, we have analyzed the structures of different substituted montmorillonite and the effect sorption behavior of Na + in different montmorillonite models. The result shows that the process of substitution will occur apart from octahedral aluminums. The adsorption of NH 3 on clay surfaces have been investigated using TPD and DFT. This is shown that acid sites locate at round the octahedral aluminums, and substitution of Al 3+ for tetrahedral Si will be favorable to NH 3 adsorption.

  15. Reconstitution of highly purified saxitoxin-sensitive Na+-channels into planar lipid bilayers.

    PubMed Central

    Hanke, W; Boheim, G; Barhanin, J; Pauron, D; Lazdunski, M

    1984-01-01

    Highly purified Na+-channels isolated from rat brain have been reconstituted into virtually solvent-free planar lipid bilayer membranes. Two different types of electrically excitable channels were detected in the absence of any neurotoxins. The activity of both channels was blocked by saxitoxin. The first channel type is highly selective for Na+ over K+ (approximately 10:1), it shows a bursting behavior, a conductance of 25 pS in Na+-Ringer and undergoes continuous opening and closing events for periods of minutes within a defined range of negative membranes voltages. The second channel type has a conductance of 150 pS and a lower selectivity for Na+ and K+ (2.2:1); only a few opening and closing events are observed with this channel after one voltage jump. The latter type of channel is also found with highly purified Na+-channel from Electrophorus electricus electroplax. A qualitative analysis of the physicochemical and pharmacological properties of the high conductance channel has been carried out. Channel properties are affected not only by saxitoxin but also by a scorpion (Centruroides suffusus suffusus) toxin and a sea anemone (Anemonia sulcata) toxin both known to be selective for the Na+-channel. The spontaneous transformation of the large conductance channel type into the small one has been considered; the two channel types may represent the expression of activity of different conformational states of the same protein. Images Fig. 1. PMID:6325173

  16. Reconstitution of highly purified saxitoxin-sensitive Na+-channels into planar lipid bilayers.

    PubMed

    Hanke, W; Boheim, G; Barhanin, J; Pauron, D; Lazdunski, M

    1984-03-01

    Highly purified Na+-channels isolated from rat brain have been reconstituted into virtually solvent-free planar lipid bilayer membranes. Two different types of electrically excitable channels were detected in the absence of any neurotoxins. The activity of both channels was blocked by saxitoxin. The first channel type is highly selective for Na+ over K+ (approximately 10:1), it shows a bursting behavior, a conductance of 25 pS in Na+-Ringer and undergoes continuous opening and closing events for periods of minutes within a defined range of negative membranes voltages. The second channel type has a conductance of 150 pS and a lower selectivity for Na+ and K+ (2.2:1); only a few opening and closing events are observed with this channel after one voltage jump. The latter type of channel is also found with highly purified Na+-channel from Electrophorus electricus electroplax. A qualitative analysis of the physicochemical and pharmacological properties of the high conductance channel has been carried out. Channel properties are affected not only by saxitoxin but also by a scorpion (Centruroides suffusus suffusus) toxin and a sea anemone (Anemonia sulcata) toxin both known to be selective for the Na+-channel. The spontaneous transformation of the large conductance channel type into the small one has been considered; the two channel types may represent the expression of activity of different conformational states of the same protein.

  17. Effects of purified herbal extract of Salvia miltiorrhiza on ischemic rat myocardium after acute myocardial infarction.

    PubMed

    Sun, Jian; Huang, Shan Hong; Tan, Benny K-H; Whiteman, Matt; Zhu, Yi Chun; Wu, Ya Jun; Ng, Yeekong; Duan, Wei; Zhu, Yi Zhun

    2005-04-29

    In the current study, we compared purified Salvia miltiorrhiza extract (PSME) with Angiotensin-converting enzyme inhibitor, Ramipril, in in vitro experiments and also in vivo using animal model of myocardial infarction. PSME was found to have a significantly higher trolox equivalent antioxidant capacity which indicated a great capacity for scavenging free radicals. PSME could also prevent pyrogallo red bleaching and DNA damage. After 2 weeks treatment with PSME or Ramipril, survival rates of rats with experimental myocardial infarction were marginally increased (68.2% and 71.4%) compared with saline (61.5%). The ratios of infarct size to left ventricular size in both PSME-and Ramipril-treated rats were significantly less than that in the saline-treated group. Activity of cardiac antioxidant enzyme superoxide dismutase (SOD) was significant higher while level of Thiobarbituric acid-reactive substances (TBARs) was lower in the PSME treated group. Purified and standardized Chinese herb could provide an alternative regimen for the prevention of ischemic heart disease.

  18. Steviol glycoside safety: are highly purified steviol glycoside sweeteners food allergens?

    PubMed

    Urban, Jonathan D; Carakostas, Michael C; Taylor, Steve L

    2015-01-01

    Steviol glycoside sweeteners are extracted from the plant Stevia rebaudiana (Bertoni), a member of the Asteraceae (Compositae) family. Many plants from this family can induce hypersensitivity reactions via multiple routes of exposure (e.g., ragweed, goldenrod, chrysanthemum, echinacea, chamomile, lettuce, sunflower and chicory). Based on this common taxonomy, some popular media reports and resources have issued food warnings alleging the potential for stevia allergy. To determine if such allergy warnings are warranted on stevia-based sweeteners, a comprehensive literature search was conducted to identify all available data related to allergic responses following the consumption of stevia extracts or highly purified steviol glycosides. Hypersensitivity reactions to stevia in any form are rare. The few cases documented in the peer-reviewed literature were reported prior to the introduction of high-purity products to the market in 2008 when many global regulatory authorities began to affirm the safety of steviol glycosides. Neither stevia manufacturers nor food allergy networks have reported significant numbers of any adverse events related to ingestion of stevia-based sweeteners, and there have been no reports of stevia-related allergy in the literature since 2008. Therefore, there is little substantiated scientific evidence to support warning statements to consumers about allergy to highly purified stevia extracts.

  19. Quantum filtering of a thermal master equation with a purified reservoir

    NASA Astrophysics Data System (ADS)

    Genoni, Marco G.; Mancini, Stefano; Wiseman, Howard M.; Serafini, Alessio

    2014-12-01

    We consider a system subject to a quantum optical master equation at finite temperature and study a class of conditional dynamics obtained by monitoring its totally or partially purified environment. More specifically, drawing from the notion that the thermal state of the environment may be regarded as the local state of a lossy and noisy two-mode squeezed state, we consider conditional dynamics ("unravellings") resulting from the homodyne detection of the two modes of such a state. Thus, we identify a class of unravellings parametrized by the loss rate suffered by the environmental two-mode state, which interpolate between direct detection of the environmental mode alone (occurring for total loss, whereby no correlation between the two environmental modes is left) and full access to the purification of the bath (occurring when no loss is acting and the two-mode state of the environment is pure). We hence show that, while direct detection of the bath is not able to reach the maximal steady-state squeezing allowed by general-dyne unravellings, such optimal values can be obtained when a fully purified bath is accessible. More generally we show that, within our framework, any degree of access to the bath purification improves the performance of filtering protocols in terms of achievable squeezing and entanglement.

  20. Reconstitution of Purified Acetylcholine Receptors with Functional Ion Channels in Planar Lipid Bilayers

    NASA Astrophysics Data System (ADS)

    Nelson, N.; Anholt, R.; Lindstrom, J.; Montal, M.

    1980-05-01

    Acetylcholine receptor, solubilized and purified from Torpedo californica electric organ under conditions that preserve the activity of its ion channel, was reconstituted into vesicles of soybean lipid by the cholate-dialysis technique. The reconstituted vesicles were then spread into monolayers at an air-water interface and planar bilayers were subsequently formed by apposition of two monolayers. Addition of carbamoylcholine caused an increase in membrane conductance that was transient and relaxed spontaneously to the base level (i.e., became desensitized). The response to carbamoylcholine was dose dependent and competitively inhibited by curare. Fluctuations of membrane conductance corresponding to the opening and closing of receptor channels were observed. Fluctuation analysis indicated a single-channel conductance of 16± 3 pS (in 0.1 M NaCl) with a mean channel open time estimated to be 35± 5 ms. Thus, purified acetylcholine receptor reconstituted into lipid bilayers exhibited the pharmacological specificity, activation, and desensitization properties expected of this receptor in native membranes.

  1. Neuropeptide Y binding sites in rat brain identified with purified neuropeptide Y-I125

    SciTech Connect

    Walker, M.W.; Miller, R.J.

    1986-03-05

    Neuropeptide Y (NPY) is a widely distributed neuronally localized peptide with 36 amino acids, 5 of which are tyrosines. The authors wished to investigate the properties of specific receptors for NPY. They therefore labeled the tyrosines with I125 using chloramine T and then purified the peptide using HPLC. A single mono-iodinated species of NPY which yielded > 85% specific binding in rat forebrain synaptosomes was selected as the ligand for all subsequent experiments. A time course of binding showed that equilibrium conditions were reached in 60 minutes at 21/sup 0/C. Scatchard plots revealed a single class of binding sites with a Kd and a Bmax of 3 x 10-10 M and 28 pmol/mg, respectively. Competition binding with unlabeled NPY showed 50% displacement of bound ligand at 1 x 10-10 M NPY. Competition binding with rat pancreatic polypeptide (RPP), a homologous peptide possessing little NPY-like activity, showed 50% displacement of bound ligand at 2 x 10/sup -7/ M RPP. No binding was observed on F-11 or PC12 neuronal cell lines, or on HSWP fibroblast cells. They conclude that NPY-I125 purified to homogeneity with HPLC is a highly selective ligand for NPY receptor sites. They are currently investigating such sites in brain, gut, and other tissues.

  2. Dataset of the proteome of purified outer membrane vesicles from the human pathogen Aggregatibacter actinomycetemcomintans.

    PubMed

    Kieselbach, Thomas; Oscarsson, Jan

    2017-02-01

    The Gram-negative bacterium Aggregatibacter actinomycetemcomitans is an oral and systemic pathogen, which is linked to aggressive forms of periodontitis and can be associated with endocarditis. The outer membrane vesicles (OMVs) of this species contain effector proteins such as cytolethal distending toxin (CDT) and leukotoxin (LtxA), which they can deliver into human host cells. The OMVs can also activate innate immunity through NOD1- and NOD2-active pathogen-associated molecular patterns. This dataset provides a proteome of highly purified OMVs from A. actinomycetemcomitans serotype e strain 173. The experimental data do not only include the raw data of the LC-MS/MS analysis of four independent preparations of purified OMVs but also the mass lists of the processed data and the Mascot.dat files from the database searches. In total 501 proteins are identified, of which 151 are detected in at least three of four independent preparations. In addition, this dataset contains the COG definitions and the predicted subcellular locations (PSORTb 3.0) for the entire genome of A. actinomycetemcomitans serotype e strain SC1083, which is used for the evaluation of the LC-MS/MS data. These data are deposited in ProteomeXchange in the public dataset PXD002509. In addition, a scientific interpretation of this dataset by Kieselbach et al. (2015) [2] is available at http://dx.doi.org/10.1371/journal.pone.0138591.

  3. The method of purifying bioengineered spider silk determines the silk sphere properties

    PubMed Central

    Jastrzebska, Katarzyna; Felcyn, Edyta; Kozak, Maciej; Szybowicz, Miroslaw; Buchwald, Tomasz; Pietralik, Zuzanna; Jesionowski, Teofil; Mackiewicz, Andrzej; Dams-Kozlowska, Hanna

    2016-01-01

    Bioengineered spider silks are a biomaterial with great potential for applications in biomedicine. They are biocompatible,biodegradable and can self-assemble into films, hydrogels, scaffolds, fibers, capsules and spheres. A novel, tag-free, bioengineered spider silk named MS2(9x) was constructed. It is a 9-mer of the consensus motif derived from MaSp2–the spidroin of Nephila clavipes dragline silk. Thermal and acidic extraction methods were used to purify MS2(9x). Both purification protocols gave a similar quantity and quality of soluble silk; however, they differed in the secondary structure and zeta potential value. Spheres made of these purified variants differed with regard to critical features such as particle size, morphology, zeta potential and drug loading. Independent of the purification method, neither variant of the MS2(9x) spheres was cytotoxic, which confirmed that both methods can be used for biomedical applications. However, this study highlights the impact that the applied purification method has on the further biomaterial properties. PMID:27312998

  4. Antifungal activity of gallic acid purified from Terminalia nigrovenulosa bark against Fusarium solani.

    PubMed

    Nguyen, Dang-Minh-Chanh; Seo, Dong-Jun; Lee, Hyang-Burm; Kim, In-Seon; Kim, Kil-Yong; Park, Ro-Dong; Jung, Woo-Jin

    2013-03-01

    The antifungal activities of methanolic extracts from Terminalia nigrovenulosa bark (TNB) was investigated for effects on the initial growth of mycelia against Fusarium solani. The ethyl acetate fraction separated from TNB demonstrated the highest antifungal activity against F. solani. The antifungal compound was isolated from TNB using silica gel column and Sephadex LH-20 chromatography combined with thin-layer chromatography and high performance liquid chromatography. Structural identification of the antifungal compound was conducted using (1)H NMR, (13)C NMR, and liquid chromatography-tandem mass spectrometry. The purified antifungal compound was gallic acid (GA) or 3,4,5-trihydroxy benzoic acid. Purified-GA possesses the high antifungal activity against F. solani, and that antifungal activity was dosage-dependent. The hyphae became collapsed and shrunken after 24 h incubation with GA (500 ppm). In pot experiments, the application of TNB crude extract was found to be effective in controlling the cucumber Fusarium root rot disease by enhancing activities of chitinase, peroxidase thereby promoting the growth of plants. The applied TNB extract significantly suppressed root rot disease compared to control. It resulted in 33, 75 and 81% disease suppression with 100, 500 and 1000 ppm of TNB crude extract, respectively. The study effectively demonstrated biological activities of the TNB extract, therefore suggesting the application of TNB for the control of soil-borne diseases of cucumber plants.

  5. Workplace performance of a loose-fitting powered air purifying respirator during nanoparticle synthesis

    NASA Astrophysics Data System (ADS)

    Koivisto, Antti J.; Aromaa, Mikko; Koponen, Ismo K.; Fransman, Wouter; Jensen, Keld A.; Mäkelä, Jyrki M.; Hämeri, Kaarle J.

    2015-04-01

    Nanoparticle (particles with diameter ≤100 nm) exposure is recognized as a potentially harmful size fraction for pulmonary particle exposure. During nanoparticle synthesis, the number concentrations in the process room may exceed 10 × 106 cm-3. During such conditions, it is essential that the occupants in the room wear highly reliable high-performance respirators to prevent inhalation exposure. Here we have studied the in-use program protection factor (PPF) of loose-fitting powered air purifying respirators, while workers were coating components with TiO2 or Cu x O y nanoparticles under a hood using a liquid flame spray process. The PPF was measured using condensation particle counters, an electrical low pressure impactor, and diffusion chargers. The room particle concentrations varied from 4 × 106 to 40 × 106 cm-3, and the count median aerodynamic diameter ranged from 32 to 180 nm. Concentrations inside the respirator varied from 0.7 to 7.2 cm-3. However, on average, tidal breathing was assumed to increase the respirator concentration by 2.3 cm-3. The derived PPF exceeded 1.1 × 106, which is more than 40 × 103 times the respirator assigned protection factor. We were unable to measure clear differences in the PPF of respirators with old and new filters, among two male and one female user, or assess most penetrating particle size. This study shows that the loose-fitting powered air purifying respirator provides very efficient protection against nanoparticle inhalation exposure if used properly.

  6. Composition and Potency Characterization of Mycobacterium avium subsp. paratuberculosis Purified Protein Derivatives

    PubMed Central

    Capsel, Randal T.; Thoen, Charles O.; Reinhardt, Timothy A.; Lippolis, John D.; Olsen, Renee; Stabel, Judith R.; Bannantine, John P.

    2016-01-01

    Mycobacterium avium subsp. paratuberculosis (MAP) purified protein derivatives (PPDs) are immunologic reagents prepared from cultured filtrates of the type strain. Traditional production consists of floating culture incubation at 37°C, organism inactivation by autoclaving, coarse filtration, and protein precipitation. Three traditional production PPDs were used in this study including lot 9801, which served as a reference and has been used in the field for decades. Alternative production PPDs (0902A and 0902B), in which the autoclaving step was removed, were also analyzed in this study. SDS-PAGE analysis revealed protein smearing in traditional PPDs, but distinct bands were observed in the alternative PPD preparations. Antibody bound distinct protein bands in the alternative PPDs by immunoblot analysis, whereas an immunoreactive smear was observed with the traditional PPDs. Mass spectrometry identified 194 proteins among three PPD lots representing the two different production methods, ten of which were present in all PPDs examined. Selected proteins identified by mass spectrometry were recombinantly expressed and purified from E. coli and evaluated by the guinea pig potency test. Seven recombinant proteins showed greater erythema as compared to the reference PPD lot 9801 in paired guinea pigs and were able to stimulate interferon-gamma production in blood from Johne’s positive animals. These results suggest that autoclaving culture suspensions is not a necessary step in PPD production and specific proteins could supplant the PPD antigen for intradermal skin testing procedures and for use as in-vitro assay reagents. PMID:27136199

  7. In vitro enhancement of human natural cell-mediated cytotoxicity by purified influenza virus glycoproteins.

    PubMed Central

    Arora, D J; Houde, M; Justewicz, D M; Mandeville, R

    1984-01-01

    The role of the glycoproteins of influenza virus, hemagglutinin (HA), and neuraminidase (NA) in the in vitro stimulation of natural cell-mediated cytotoxicity (NCMC) or natural killer activity of human peripheral blood lymphocytes was evaluated with radiolabeled K562 cells as target cells in an overnight chromium release assay. Three different approaches were used. (i) Purified viral proteins were obtained by extraction with Nonidet P-40, separation on a sucrose gradient, and further purification by affinity chromatography. Ficoll-Hypaque-purified peripheral blood lymphocytes exposed to HA or NA individually or to a mixture of both significantly increased NCMC (32 to 50%). (ii) Treatment of HA and NA with their respective homologous antisera or F(ab')2 antibody abrogated the stimulation of NCMC by these glycoproteins. (iii) Virions treated with proteolytic enzymes resulted in viral cores lacking either HA or NA or both activities. Compared to whole virions, viral cores devoid of HA activity only induced a 50% increase in NCMC, whereas viral cores lacking HA activity and with traces of NA activity stimulated only 10% of the NCMC. These results suggest that influenza virus-induced cell-mediated cytotoxicity is largely due to its glycoproteins. PMID:6387178

  8. Purified human platelet-derived growth factor receptor has ligand-stimulated tyrosine kinase activity.

    PubMed Central

    Bishayee, S; Ross, A H; Womer, R; Scher, C D

    1986-01-01

    The platelet-derived growth factor receptor (PDGF-R), a 180-kDa single-chain polypeptide, was purified from membranes of the human osteogenic sarcoma cell line MG-63. Purification was achieved by treatment of membranes with PDGF and ATP, followed by solubilization with nonionic detergent and successive chromatography on solid-phase anti-phosphotyrosine monoclonal antibody and DEAE-cellulose. The PDGF-R, which was estimated to be 50-80% pure by NaDodSO4/polyacrylamide gel electrophoresis of 32P-labeled preparations, was free of contaminating epidermal growth factor receptor and had no detectable phosphatase activity. It specifically bound 125I-labeled PDGF, a reaction quantified by binding of the ligand-PDGF-R complex to the anti-phosphotyrosine antibody. The purified receptor displayed PDGF-stimulatable tyrosine kinase activity, assayed by autophosphorylation of PDGF-R at tyrosine residues and by phosphorylation of angiotensin II. The Km for ATP in the autophosphorylation reaction was 7.5 microM. Addition of PDGF did not change the Km but increased the Vmax 1.7-fold. Images PMID:3018745

  9. Stability of serum-free and purified baculovirus stocks under various storage conditions.

    PubMed

    Jorio, Hasnaa; Tran, Rosa; Kamen, Amine

    2006-01-01

    In a context of large-scale production of baculoviruses in serum-free media for use as gene delivery vectors, the stability of these viruses has become an important factor. The development of robust processes heavily relies on baculovirus stock stability. In the present work, we studied over a period of 300 days the stability of baculovirus vectors produced in serum-free media stored at 4, -20, or -80 degrees C or in liquid nitrogen. The viral stocks investigated were either crude baculovirus supernatant, baculovirus supernatant concentrated 10 times and diafiltered against fresh serum-free media by tangential flow filtration, or baculovirus purified by size exclusion chromatography. The results showed that baculovirus supernatant and diafiltered concentrate stored at 4 degrees C underwent a progressive loss of infectivity after a period of 100 and 50 days of storage, respectively. Aggregation has been recognized as the probable mechanism for the loss of infectivity. Baculovirus stocks were unstable at -20 degrees C, whereas in liquid nitrogen they retained infectivity after successive freeze thaw cycles. Concentration and diafiltration of baculovirus supernatant prior to storing at -80 degrees C contributed to improving viral stock stability over time. Glycerol as well as DMSO and sucrose have proven to be equally effective as additives to maintain the purified baculovirus stability after storage at -80 degrees C or in liquid nitrogen.

  10. Production of Self-Purifying Proteins in a Variety of Expression Hosts with Focus on Organophosphorus Hydrolase

    DTIC Science & Technology

    2012-08-17

    REPORT Final Report: Production of Self-Purifying Proteins in a Variety of Expression Hosts with Focus on Organophosphorus Hydrolase 14. ABSTRACT 16...Prescribed by ANSI Std. Z39.18 - 31-Mar-2012 Final Report: Production of Self-Purifying Proteins in a Variety of Expression Hosts with Focus on...highly effective methods for protein purification. We have applied these methods to the production of several proteins, including

  11. The 73 kilodalton heat shock cognate protein purified from rat brain contains nonesterified palmitic and stearic acids.

    PubMed

    Guidon, P T; Hightower, L E

    1986-08-01

    A protein related to the 71 kilodalton inducible rat heat shock protein was purified to electrophoretic homogeneity in milligram amounts from brain tissue of nonheat-stressed rats. The protein has been designated as a stress cognate protein based on previous studies and data presented herein that this protein cross-reacted with a monoclonal antibody originally raised against the Drosophila 70 kilodalton heat shock protein. The purified protein had an apparent molecular mass of 73 kilodaltons when analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and an apparent mass of 150 kilodaltons as determined by nondissociative gel chromatography, suggesting that the purified protein is a homodimer. The purified protein had isoelectric points of 5.0 under nondissociative conditions and 5.6 when exposed to protein denaturants, suggesting loss of bound anionic molecules and/or net exposure of basic residues upon denaturation. Chloroform/methanol extraction of the purified protein and subsequent analyses by thin layer and gas-liquid chromatography resulted in the identification of palmitic and stearic acids noncovalently bound to the protein. Approximately four molecules of fatty acids were bound per dimer with palmitic and stearic acids present in a one-to-one ratio. The purified protein did not bind exogenously added radioactive palmitate, indicating that the fatty acid-binding sites of the cognate protein were fully occupied and that the associated fatty acids were too tightly bound to exchange readily. The possible significance of the fatty acids associated with the 73 kilodalton stress cognate protein is discussed.

  12. Characterization of a purified nicotinic receptor from rat brain by using idiotypic and anti-idiotypic antibodies

    SciTech Connect

    Abood, L.G.; Langone, J.J.; Bjercke, R.; Lu, X.; Banerjee, S.

    1987-09-01

    The availability of an anti-nicotine monoclonal antibody has made it possible to further establish the nature of the nicotine recognition proteins purified from rat brain by affinity chromatography and to provide a highly sensitive assay for determining (/sup 3/H)nicotine binding to the purified material. An enantiomeric analogue of nicotine. (-)-6-hydroxymethylnicotine, was used to prepare the affinity column. In addition, with the use of an anti-idiotypic monoclonal antibody, it was confirmed that the recognition site for nicotine resides on a protein complex composed of two components with molecular masses of 62 and 57 kDa. It was also demonstrated that the same two proteins could be purified by immunoaffinity chromatography with the use of an anti-idiotypic monoclonal antibody. With the use of the anti-nicotine antibody to measure (/sup 3/H)nicotine binding, the purified material was shown to bind 250 pmol/mg of protein. By utilizing a procedure in which the purified receptor protein was conjugated to membranes by disulfide bonds, a binding activity of 80 pmol/mg was obtained. With the availability of sterospecific monoclonal antibodies to (-)-nicotine as well as monoclonal anti-idiotypic antibodies derived when the anti-nicotine antibodies were used as immunogens, additional procedures became available for the further characterization of the purified nicotine receptor and examining its (-)-(/sup 3/H)nicotine-binding characteristics.

  13. Demonstration of protein-fragment complementation assay using purified firefly luciferase fragments

    PubMed Central

    2013-01-01

    Background Human interactome is predicted to contain 150,000 to 300,000 protein-protein interactions, (PPIs). Protein-fragment complementation assay (PCA) is one of the most widely used methods to detect PPI, as well as Förster resonance energy transfer (FRET). To date, successful applications of firefly luciferase (Fluc)-based PCA have been reported in vivo, in cultured cells and in cell-free lysate, owing to its high sensitivity, high signal-to-background (S/B) ratio, and reversible response. Here we show the assay also works with purified proteins with unexpectedly rapid kinetics. Results Split Fluc fragments both fused with a rapamycin-dependently interacting protein pair were made and expressed in E. coli system, and purified to homogeneity. When the proteins were used for PCA to detect rapamycin-dependent PPI, they enabled a rapid detection (~1 s) of PPI with high S/B ratio. When Fn7-8 domains (7 nm in length) that was shown to abrogate GFP mutant-based FRET was inserted between split Fluc and FKBP12 as a rigid linker, it still showed some response, suggesting less limitation in interacting partner’s size. Finally, the stability of the probe was investigated. Preincubation of the probes at 37 degreeC up to 1 h showed marked decrease of the luminescent signal to 1.5%, showing the limited stability of this system. Conclusion Fluc PCA using purified components will enable a rapid and handy detection of PPIs with high S/B ratio, avoiding the effects of concomitant components. Although the system might not be suitable for large-scale screening due to its limited stability, it can detect an interaction over larger distance than by FRET. This would be the first demonstration of Fluc PCA in vitro, which has a distinct advantage over other PPI assays. Our system enables detection of direct PPIs without risk of perturbation by PPI mediators in the complex cellular milieu. PMID:23536995

  14. Sugar binding to purified fractions from bovine taste buds and epithelial tissue. Relationships to bioactivity.

    PubMed

    Lum, C K; Henkin, R I

    1976-02-24

    Binding of various sugars was compared in purified subfractions of taste buds isolated from bovine circumvallate papillae and of non-taste bud-bearing epithelium isolated from tissue surrounding these papillae. Binding of 14C-labeled sugars was greater in purified subfractions obtained from taste bud than from non-taste bud-bearing tissue and was, in general, greater in those taste bud subfractions in which a greater membrane purification was achieved. Binding specificity of the 14C-labeled sugars sucrose, fructose, glucose and of 14C-labeled cyclamate and saccharine was measured by competition of each 14C-labeled sugar or synthetic sweetener with its unlabeled homologous sugar in P4(B) taste bud subfractions; this binding, as shown for sucrose, was reversible and temperature dependent. Essentially no competition of the 14C-lageled sugars sucrose, fructose, glucose or 14C-labeled cyclamate and saccharine by their respective unlabeled homologues occurred in epithelial tissue P4(B) subfractions; this binding was not reversible. Binding specificity was further observed by the competition of 14C-labeled sucrose, fructose and glucose with each unlabeled sugar for binding sites on P4(B) taste bud subfractions; unlabeled sucrose was more effective in competing with each 14C-labeled surgar than was unlabeled fructose or glucose. The relatively non-sweet sugar lactose did not compete with 14C-labeled lactose in P4(B) subfractions from either taste bud or non-taste bud-bearing epithelial tissue. Binding of 14C-labeled sucrose in purified P4(B) bud subfractions was inhibited by increased concentrations of unlabeled sucrose, phospholipase C, neuraminidase, EDTA, NaCl and urea. Dissociation constants for sugar or synthetic sweetener binding were low (approx. 10(-3) M) but in a rank order (sucrose greater than fructose greater than glucose greater than saccharine) consistent with preference and electrophysiological responses in cow. The cow is behaviorally indifferent to

  15. Evaluation of cross-linked aggregates from purified Bacillus subtilis levansucrase mutants for transfructosylation reactions

    PubMed Central

    Ortiz-Soto, Maria Elena; Rudiño-Piñera, Enrique; Rodriguez-Alegria, Maria Elena; Munguia, Agustin Lopez

    2009-01-01

    Background Increasing attention has been focused on inulin and levan-type oligosaccharides, including fructosyl-xylosides and other fructosides due to their nutraceutical properties. Bacillus subtilis levansucrase (LS) catalyzes the synthesis of levan from sucrose, but it may also transfer the fructosyl moiety from sucrose to acceptor molecules included in the reaction medium. To study transfructosylation reactions with highly active and robust derivatives, cross-linked enzyme aggregates (CLEAs) were prepared from wild LS and two mutants. CLEAs combine the catalytic features of pure protein preparations in terms of specific activity with the mechanical behavior of industrial biocatalysts. Results Two types of procedures were used for the preparation of biocatalysts from purified wild type LS (WT LS) B. subtilis and the R360K and Y429N LS mutants: purified enzymes aggregated with glutaraldehyde (cross-linked enzyme aggregates: CLEAs), and covalently immobilized enzymes in Eupergit C®. The biocatalysts were characterized and used for fructoside synthesis using xylose as an acceptor model. CLEAs were able to catalyze the synthesis of fructosides as efficiently as soluble enzymes. The specific activity of CLEAs prepared from wild type LS (44.9 U/mg of CLEA), R360K (56.5 U/mg of CLEA) and Y429N (1.2 U/mg of CLEA) mutants were approximately 70, 40 and 200-fold higher, respectively, than equivalent Eupergit C® immobilized enzyme preparations (U/mg of Eupergit), where units refer to global LS activity. In contrast, the specific activity of the free enzymes was 160, 171.2 and 1.5 U/mg of protein, respectively. Moreover, all CLEAs had higher thermal stability than corresponding soluble enzymes. In the long term, the operational stability was affected by levan synthesis. Conclusion This is the first report of cross-linked transglycosidases aggregates. CLEAs prepared from purified LS and mutants have the highest specific activity for immobilized fructosyltransferases (FTFs

  16. IDENTIFICATION AND CHARACTERIZATION OF INFECTIOUS AND NON-INFECTIOUS SUB-POPULATIONS OF ENCEPHALITIZOON INTESTINALIS SPORES PURIFIED FROM IN VITRO CELL CULTURE

    EPA Science Inventory

    Background: Encephalitizoon intestinalis spores were propagated in rabbit kidney (RK-13) cells and were purified using density gradient (Percoll [registered trademark]) centrifugation. Purified spores were enumeraged and aliquotted using flow cytometry with cell sorting for use...

  17. Use of the Meissner effect to separate, purify, and classify superconducting powders

    SciTech Connect

    Barsoum, M.; Patten, D.; Tyagi, S.

    1987-12-07

    A separation technique, based on the Meissner effect, to purify and classify superconducting powders was developed. Powders of the Y/sub 1/Ba/sub 2/Cu/sub 3/O/sub 7-//sub x/ composition were sealed in a glass tube, cooled to liquid-nitrogen temperature, and passed over the pole of a magnet. The superconducting phase responded to the field and moved away from the magnet, whereas the nonsuperconducting phases, impurities, especially magnetic, and unreacted phases did not move. X-ray diffraction revealed that the separated powder was single phase with all impurity phases eliminated. Furthermore, the powder particles separated along the length of the tube according to their size and applied magnetic field: the larger the particles and/or the field the farther the movement and the smaller the particles that moved.

  18. Antiviral activities of purified compounds from Youngia japonica (L.) DC (Asteraceae, Compositae).

    PubMed

    Ooi, Linda S M; Wang, Hua; He, Zhendan; Ooi, Vincent E C

    2006-06-30

    The ethanol extract of a biannual medicinal herb, Youngia japonica (commonly known as Oriental hawk's beard) was reported previously to have potent antiviral activity against respiratory syncytial virus (RSV) cultured in HEp-2 cells. Three anti-microbial agents, namely 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, and luteolin-7-O-glucoside were subsequently purified and chemically characterized from the ethanol extract of Youngia japonica. The two dicaffeoylquinic acids exhibited prominent anti-RSV with 50% inhibitory concentration (IC50) of 0.5 microg/ml in vitro. Luteolin-7-O-glucoside together with the two dicaffeoylquinic acids were also manifested to have some antibacterial activity towards the causal agents of food-borne disease, namely Vibrio cholerae and Vibrio parahaemolyticus at the concentration of 2mg/ml. Bacillus cereus was sensitive to 3,4-dicaffeoylquinic acid and 3,5-dicaffeoylquinic acid only, but not to luteolin-7-O-glucoside.

  19. Inter-comparison in 10Be analysis starting from pre-purified quartz

    NASA Astrophysics Data System (ADS)

    Schnabel, C.; Reinhardt, L.; Barrows, T. T.; Bishop, P.; Davidson, A.; Fifield, L. K.; Freeman, S.; Kim, J. Y.; Maden, C.; Xu, S.

    2007-06-01

    The results of the first international inter-comparison of 10Be analysis from quartz are presented. This inter-comparison includes the sample preparation starting from pre-purified quartz and AMS measurements at SUERC and ANU. Measured 10Be concentrations agree within their uncertainties for six out of seven samples with 10Be concentrations greater than 1 × 104 at/g quartz. This agreement and also the agreement of 10Be concentrations analysed from two aliquots of the same sample at SUERC indicate that addition of 9Be carrier before (used at ANU) or after quartz dissolution (used at SUERC apart from one aliquot of one sample) should not result in substantially different results.

  20. A simple filtration technique for obtaining purified human chromosomes in suspension.

    PubMed

    Yusuf, Mohammed; Parmar, Neha; Bhella, Gurdeep K; Robinson, Ian K

    2014-05-01

    Here we present a simple method for cleaning polyamine human mitotic chromosomes in solution. This was achieved by filtering intact (unburst) nuclei along with both large and small cytoplasmic debris through a series of different pore sized filters. Pure human chromosomes were recovered using a simple reverse filtration step. Fluorescence microscopy was used to validate the chromosome suspension after each filtration step. This reverse filtration technique is an improvement in both procedure time and chromosome recovery compared to currently used post-purification methods. Chromosomes purified by our method could be used for many applications, such as structural studies using microfluidics and high resolution imaging or generation of chromosome paints and sequencing after flow cytometry.

  1. Use of collagenase to purify collagen from prehistoric bones for stable isotopic analysis

    NASA Astrophysics Data System (ADS)

    DeNiro, Michael J.; Weiner, Stephen

    1988-10-01

    The low molecular weight hydrolysis products of the reaction between collagenase and the HCl insoluble organic fraction from modern and well-preserved prehistoric bones have the same stable carbon and nitrogen isotopic compositions and amino acid composition as "collagen" extracted by dissolution in warm dilute acid. The hydrolysis products produced using the collagenase method from some poorly preserved fossil bones have the same amino acid composition as the products produced from modern bones, even though the "collagen" extracted from these bones by the conventional solubility method bears no similarity to modern collagen either in terms of amino acid or isotopic composition. These observations suggest that collagenase can possibly be used to purify a fraction from such poorly preserved prehistoric bones that retains its in vivo isotope ratios, thereby permitting isotopic paleodietary reconstruction for bones that cannot be studied with currently available techniques.

  2. Effect immunization with highly purified alpha- and beta-toxins on staphylococcal mastitis in rabbits.

    PubMed

    Adlam, C; Ward, P D; McCartney, A C; Arbuthnott, J P; Thorley, C M

    1977-08-01

    Experiments were carried out to determine whether immunization of female rabbits with highly purified staphylococcal alpha- or beta-toxins would protect them against intramammary challenge with staphylococci. High circulating anti-alpha-toxin titers reduced the lethal hemorrhagic edematous form of the disease ("blue-breast") produced by strains BB and Compton 201 to a localized chronic abscess form. No such protection was afforded by high anti-beta-toxin titers. Immunization with alpha- or beta-toxins produced no change in the clinical picture of the disease produced by CN.6708, a strain of Staphylococcus responsible for a natural outbreak of abscess-type rabbit mastitis. From these experiments it would appear that alpha-toxin is a key antigen in the blue-breast form of rabbit mastitis. Since the abscess form of the disease was not prevented by immunization with either alpha- or beta-toxin, other virulence factors must be acting to produce this more localized disease.

  3. Affinity chromatography approaches to overcome the challenges of purifying plasmid DNA.

    PubMed

    Sousa, Fani; Prazeres, Duarte M F; Queiroz, João A

    2008-09-01

    The diversity of biomolecules present in plasmid DNA (pDNA)-containing extracts and the structural and chemical similarities between pDNA and impurities are some of the main challenges of improving or establishing novel purification procedures. In view of the unequalled specificity of affinity purification, this technique has recently begun to be applied in downstream processing of plasmids. This paper discusses the progress and importance of affinity chromatography (AC) for the purification of pDNA-based therapeutic products. Several affinity approaches have already been successfully developed for a variety of applications, and we will focus here on highlighting their possible contributions to the pDNA purification challenge. Diverse affinity applications and their advantages and disadvantages are discussed, as well as the most significant results and improvements in the challenging task of purifying plasmids.

  4. Multiple specific binding sites for purified glucocorticoid receptors on mammary tumor virus DNA.

    PubMed

    Payvar, F; Firestone, G L; Ross, S R; Chandler, V L; Wrange, O; Carlstedt-Duke, J; Gustafsson, J A; Yamamoto, K R

    1982-01-01

    Glucocorticoid hormones selectively stimulate the rate of transcription of integrated mammary tumor virus (MTV) sequences in infected rat hepatoma cells. Using two independent assays, we find that purified rat liver glucocorticoid receptor protein binds specifically to at least four widely separated regions on pure MTV proviral DNA. One of these specific binding domains, which itself contains at least two distinct receptor binding sites, resides within a fragment of viral DNA that maps 110-449 bp upstream of the promoter for MTV RNA synthesis. Three other binding domains lie downstream of the promoter and within the MTV primary transcription unit. Restriction fragments bearing separate binding domains have been introduced into cultured cells; transformants have been recovered in which the introduced fragments are expressed under glucocorticoid control. Thus, it appears that this assay will be useful for assessing the biological significance of the receptor binding sites detected in vitro.

  5. Molecular and functional characteristics of purified gum from Australian chia seeds.

    PubMed

    Timilsena, Yakindra Prasad; Adhikari, Raju; Kasapis, Stefan; Adhikari, Benu

    2016-01-20

    Chia seed gum (CSG) was extracted from the seed coat of Salvia hispanica, purified in the laboratory and its chemical composition and functional properties were investigated. CSG was found to comprise 93.8% carbohydrate consisting of xylose, glucose, arabinose, galactose, glucuronic acid and galacturonic acid as monosaccharide units. The presence of uronic acids was reflected in the anionic behavior of the CSG solution over a wide range of pH (≥ 1.8). The solubility of CSG increased slightly with temperature and pH of the aqueous medium. CSG was able to resist pyrolytic decomposition at temperatures well in excess of 250 °C, and exhibited a high water holding capacity (23 times of its own weight). The surface activity and emulsifying properties of CSG were found to be either superior or comparable to other common gums and industrial polysaccharides indicating the potential of CSG as an effective thickener and stabilizer of processed foods.

  6. Several properties of the partially purified proteinase inhibitor in eggplant exocarp.

    PubMed

    Kanamori, M; Ibuki, F; Yamada, M; Tashiro, M; Miyoshi, M

    1975-01-01

    A proteinase inhibitor was isolated and partially purified from the exocarp of eggplant, Solanum melongena L., by means of acetate buffer extraction, heat treatment, salting-out and column chromatography on DEAE-cellulose. This preparation showed inhibitory activities on various proteinases; trypsin [EC 3.4.4.4] and Pronase were strongly inhibited while alpha-chymotrypsin [EC 3.4.4.5] and Nagarse were weakly inhibited. The inhibitor was a protein substance, and, therefore, it was gradually inactivated by the long-time incubation with Pronase. The inhibition mode was non-competitive on trypsin and competitive on Pronase on the basis of Lineweaver-Burk plots. The investigations on the inhibition behavior in the co-existence of two kinds of proteinases suggested that the inhibitor was not of multi-headed type.

  7. Protein-protein interactions of tandem affinity purified protein kinases from rice.

    PubMed

    Rohila, Jai S; Chen, Mei; Chen, Shuo; Chen, Johann; Cerny, Ronald L; Dardick, Christopher; Canlas, Patrick; Fujii, Hiroaki; Gribskov, Michael; Kanrar, Siddhartha; Knoflicek, Lucas; Stevenson, Becky; Xie, Mingtang; Xu, Xia; Zheng, Xianwu; Zhu, Jian-Kang; Ronald, Pamela; Fromm, Michael E

    2009-08-19

    Eighty-eight rice (Oryza sativa) cDNAs encoding rice leaf expressed protein kinases (PKs) were fused to a Tandem Affinity Purification tag (TAP-tag) and expressed in transgenic rice plants. The TAP-tagged PKs and interacting proteins were purified from the T1 progeny of the transgenic rice plants and identified by tandem mass spectrometry. Forty-five TAP-tagged PKs were recovered in this study and thirteen of these were found to interact with other rice proteins with a high probability score. In vivo phosphorylated sites were found for three of the PKs. A comparison of the TAP-tagged data from a combined analysis of 129 TAP-tagged rice protein kinases with a concurrent screen using yeast two hybrid methods identified an evolutionarily new rice protein that interacts with the well conserved cell division cycle 2 (CDC2) protein complex.

  8. Protein-Protein Interactions of Tandem Affinity Purified Protein Kinases from Rice

    PubMed Central

    Rohila, Jai S.; Chen, Mei; Chen, Shuo; Chen, Johann; Cerny, Ronald L.; Dardick, Christopher; Canlas, Patrick; Fujii, Hiroaki; Gribskov, Michael; Kanrar, Siddhartha; Knoflicek, Lucas; Stevenson, Becky; Xie, Mingtang; Xu, Xia; Zheng, Xianwu; Zhu, Jian-Kang; Ronald, Pamela; Fromm, Michael E.

    2009-01-01

    Eighty-eight rice (Oryza sativa) cDNAs encoding rice leaf expressed protein kinases (PKs) were fused to a Tandem Affinity Purification tag (TAP-tag) and expressed in transgenic rice plants. The TAP-tagged PKs and interacting proteins were purified from the T1 progeny of the transgenic rice plants and identified by tandem mass spectrometry. Forty-five TAP-tagged PKs were recovered in this study and thirteen of these were found to interact with other rice proteins with a high probability score. In vivo phosphorylated sites were found for three of the PKs. A comparison of the TAP-tagged data from a combined analysis of 129 TAP-tagged rice protein kinases with a concurrent screen using yeast two hybrid methods identified an evolutionarily new rice protein that interacts with the well conserved cell division cycle 2 (CDC2) protein complex. PMID:19690613

  9. Positive and strongly relaxed purifying selection drive the evolution of repeats in proteins

    PubMed Central

    Persi, Erez; Wolf, Yuri I.; Koonin, Eugene V

    2016-01-01

    Protein repeats are considered hotspots of protein evolution, associated with acquisition of new functions and novel phenotypic traits, including disease. Paradoxically, however, repeats are often strongly conserved through long spans of evolution. To resolve this conundrum, it is necessary to directly compare paralogous (horizontal) evolution of repeats within proteins with their orthologous (vertical) evolution through speciation. Here we develop a rigorous methodology to identify highly periodic repeats with significant sequence similarity, for which evolutionary rates and selection (dN/dS) can be estimated, and systematically characterize their evolution. We show that horizontal evolution of repeats is markedly accelerated compared with their divergence from orthologues in closely related species. This observation is universal across the diversity of life forms and implies a biphasic evolutionary regime whereby new copies experience rapid functional divergence under combined effects of strongly relaxed purifying selection and positive selection, followed by fixation and conservation of each individual repeat. PMID:27857066

  10. Dynamics of spherical metallic particles in cylinder electrostatic separators/purifiers.

    PubMed

    Lu, Hong-Zhou; Li, Jia; Guo, Jie; Xu, Zhen-Ming

    2008-08-15

    This paper presents a theoretical analysis of the dynamics of spherical metallic particles in electrostatic separators/purifiers (ESPs). The particle equations of motion are numerically solved in two dimensions using a computational algorithm. The ESPs consist of a pair of conductor cylinder electrodes. The upper cylinder is energized by HVdc, while the lower one is grounded and fixed horizontally on a revolvable axis. Some phenomena and aspects of separation process are explained and depicted including lifting off, impact, "motion collapse" and "sudden bouncing". The results reveal that the several phenomena depend on initial position, radius and density of the particle, curvature of the cylinder electrodes, distance between the electrodes and amplitude of the applied voltage. Optimization of the parameters is presented in order to get better separation/purification processes.

  11. Electrochemical Behavior of Titanium(II) Ion in a Purified Calcium Chloride Melt

    NASA Astrophysics Data System (ADS)

    Kang, Min Ho; Song, Jianxun; Zhu, Hongmin; Jiao, Shuqiang

    2014-09-01

    Cyclic voltammetry, chronopotentiometry, and square wave voltammetry were used to investigate electrochemical behavior of Ti(II) ion in a purified CaCl2 melt at a temperature of 1173 K (900 °C). The result indicated that the cathodic reduction of Ti(II) ion in the melt was a one-step quasi-reversible process controlled by the diffusion. The diffusion coefficient was determined in a CaCl2-TiCl(0.5 mol/dm3) at 1173 K (900 °C). The work also demonstrated the feasibility of producing metallic titanium in the as-prepared CaCl2-TiCl2 melts through galvanostatic electrolysis.

  12. Electrochemical Behavior of Titanium(II) Ion in a Purified Calcium Chloride Melt

    NASA Astrophysics Data System (ADS)

    Kang, Min Ho; Song, Jianxun; Zhu, Hongmin; Jiao, Shuqiang

    2015-02-01

    Cyclic voltammetry, chronopotentiometry, and square wave voltammetry were used to investigate electrochemical behavior of Ti(II) ion in a purified CaCl2 melt at a temperature of 1173 K (900 °C). The result indicated that the cathodic reduction of Ti(II) ion in the melt was a one-step quasi-reversible process controlled by the diffusion. The diffusion coefficient was determined in a CaCl2-TiCl(0.5 mol/dm3) at 1173 K (900 °C). The work also demonstrated the feasibility of producing metallic titanium in the as-prepared CaCl2-TiCl2 melts through galvanostatic electrolysis.

  13. Retinoblastoma protein co-purifies with proteasomal insulin-degrading enzyme: Implications for cell proliferation control

    SciTech Connect

    Radulescu, Razvan T.; Duckworth, William C.; Levy, Jennifer L.; Fawcett, Janet

    2010-04-30

    Previous investigations on proteasomal preparations containing insulin-degrading enzyme (IDE; EC 3.4.24.56) have invariably yielded a co-purifying protein with a molecular weight of about 110 kDa. We have now found both in MCF-7 breast cancer and HepG2 hepatoma cells that this associated molecule is the retinoblastoma tumor suppressor protein (RB). Interestingly, the amount of RB in this protein complex seemed to be lower in HepG2 vs. MCF-7 cells, indicating a higher (cytoplasmic) protein turnover in the former vs. the latter cells. Moreover, immunofluorescence showed increased nuclear localization of RB in HepG2 vs. MCF-7 cells. Beyond these subtle differences between these distinct tumor cell types, our present study more generally suggests an interplay between RB and IDE within the proteasome that may have important growth-regulatory consequences.

  14. Nitric Oxide Measurement from Purified Enzymes and Estimation of Scavenging Activity by Gas Phase Chemiluminescence Method.

    PubMed

    Kumari, Aprajita; Gupta, Alok Kumar; Mishra, Sonal; Wany, Aakanksha; Gupta, Kapuganti Jagadis

    2016-01-01

    In plants, nitrate reductase (NR) is a key enzyme that produces nitric oxide (NO) using nitrite as a substrate. Lower plants such as algae are shown to have nitric oxide synthase enzyme and higher plants contain NOS activity but enzyme responsible for NO production in higher plants is subjected to debate. In plant nitric oxide research, it is very important to measure NO very precisely in order to determine its functional role. A significant amount of NO is being scavenged by various cell components. The net NO production depends in production minus scavenging. Here, we describe methods to measure NO from purified NR and inducible nitric oxide synthase from mouse (iNOS), we also describe a method of measure NO scavenging by tobacco cell suspensions and mitochondria from roots.

  15. Fused salt process for purifying zirconium and/or hafnium tetrachlorides

    SciTech Connect

    Lee, E.D.

    1991-04-23

    This patent describes a fused salt process for continuously purifying zirconium and/or hafnium tetrachloride dissolved in a molten bath in a vessel. It comprises: maintaining a mass of a suitable mixture of salts, including zirconium and/or hafnium tetrachloride; heating the mixture of salts to a temperature at or immediately below the vaporization temperature of the zirconium and/or hafnium tetrachloride at which temperature the mixture of salts is fused to form a molten, tetrachloride-dissolving bath; continuously introducing into the dissolving bath a zirconium and/or hafnium tetrachloride powder; heating a portion of the dissolving bath in situ to a temperature higher than the vaporization temperature of the zirconium and/or hafnium tetrachloride so as to vaporize the tetrachloride; internally circulating the dissolving bath whereby the portion of the dissolving bath at the high temperature circulate with the bath at the lower temperature.

  16. Investigation of durability of optical coatings in highly purified tritium gas

    SciTech Connect

    Fischer, S.; Schoenung, K.; Bornschein, B.; Rolli, R.; Schaefer, V.; Sturm, M.

    2015-03-15

    Anti-reflection coated windows are part of Raman spectroscopy systems for tritium analytics in the KATRIN experiment and fusion-related applications. Damages of such windows were observed after three months of expo-sure to highly purified tritium gas in the LOOPINO facility. In this work, the origin of the damages was investigated, identified and eliminated. Coating samples manufactured by various physical vapor deposition methods have been tested for durability by exposure to pure tritium gas and subsequent visual inspection. Electron beam deposited coatings showed indications for damage after 17 days of tritium exposure in contrast to samples manufactured by ion assisted deposition or sputtering. An improved coating layout of the sample cell is presented for reliable long-term monitoring of tritium gas using Raman spectroscopy. (authors)

  17. Purified recombinant organophosphorus acid anhydrase protects mice against soman. (Reannouncement with new availability information)

    SciTech Connect

    Broomfield, C.A.

    1992-12-31

    Since pharmacologic treatments of organophosphorus anticholinesterases (OPs) are nearing their practical limit other types of treatment are being sought. One approach is the prophylactic administration of scavengers that will detoxify OPs before they reach their critical target site. Using mice that were sensitized to OPs by depletion of their serum carboxylesterase with cresylbenzodioxaphosphorin oxide (CBDP), we have shown that animals pretreated intravenously with a purified organophosphorus acid anhydride hydrolase (parathionase) (0.10 mg per g body wt.) are not measurably affected by up to 34.4 microgram soman per kg, a dose more than double that which is lethal to untreated animals. This result indicates that this approach is worthy of exploration and development for protecting military personnel and agricultural workers against OP intoxication. Scavengers, pretreatment, soman, OP intoxication, mice.

  18. The aetiology of wobbly possum disease: Reproduction of the disease with purified nidovirus.

    PubMed

    Giles, Julia; Perrott, Matthew; Roe, Wendi; Dunowska, Magdalena

    2016-04-01

    The objective of this study was to investigate a role of a recently discovered marsupial nidovirus in the development of a neurological disease, termed wobbly possum disease (WPD), in the Australian brushtail possum (Trichosurus vulpecula). Four possums received 1 mL of a standard inoculum that had been prepared from tissues of WPD-affected possums, 4 possums received 1.8 mL (1 × 10(6) TCID50) of a cell lysate from inoculated cultures, and 4 possums received 1 mL (× 10(7) TCID50) of a purified WPD isolate. All but one possum that received infectious inocula developed neurological disease and histopathological lesions characteristic for WPD. High levels of viral RNA were detected in livers from all possums that received infectious inocula, but not from control possums. Altogether, our data provide strong experimental evidence for the causative involvement of WPD virus in development of a neurological disease in infected animals.

  19. Polysaccharides purified from Cordyceps cicadae protects PC12 cells against glutamate-induced oxidative damage.

    PubMed

    Olatunji, Opeyemi J; Feng, Yan; Olatunji, Oyenike O; Tang, Jian; Wei, Yuan; Ouyang, Zhen; Su, Zhaoliang

    2016-11-20

    Two polysaccharides CPA-1 and CPB-2 were isolated purified from Cordyceps cicadae by hot water extraction, ethanol precipitation and purification using anion exchange and gel filtration chromatography. Preliminary structural characterization of CPA-1 and CPB-2 were performed. The protective effect of CPA-1 and CPB-2 against glutamate-induced oxidative toxicity in PC12 cells was analyzed. The results indicated that pretreatment of PC12 cells with CPA-1 and CPB-2 significantly increased cell survival, Ca(2+) overload and ROS generation. CPA-1 and CPB-2 also markedly up-regulated the antioxidant status of pretreated PC12 cells. Our results suggested that Cordyceps cicadae polysaccharides can protect PC12 cells against glutamate excitotoxicity and might serve as therapeutic agents for neuronal disorders.

  20. Biochemical characteristics and antioxidant activity of crude and purified sulfated polysaccharides from Gracilaria fisheri.

    PubMed

    Imjongjairak, Siriluck; Ratanakhanokchai, Khanok; Laohakunjit, Natta; Tachaapaikoon, Chakrit; Pason, Patthra; Waeonukul, Rattiya

    2016-01-01

    Sulfated polysaccharides (SPs) from Gracilaria fisheri of Thailand, which were extracted in low-temperature (25 °C) water showed the highest content of phenolic compounds compared with those extracted at high temperature (55 °C). Crude SP antioxidant activity was evaluated by measuring the DPPH free radical scavenging effect which is directly related to the level of phenolic compounds. The sulfate content, total sugar, and SPs yield were also directly related to the extraction temperature. All extracts contained galactose as a major monosaccharide. High antioxidant activity of crude SP, positively correlated with the phenolic compound contents (R(2) = 0.996) contributed by the existence of sulfate groups and phenolic compounds. In purified SP, F1 fraction exhibited strong radical scavenging ability, but it was not significantly different compared to crude SP extracted at 25 °C. This indicated that the appropriate density and distribution of sulfate groups in the SP extract showed the best antioxidant activity.

  1. Immobilized purified folate-binding protein: binding characteristics and use for quantifying folate in erythrocytes

    SciTech Connect

    Hansen, S.I.; Holm, J.; Nexo, E.

    1987-08-01

    Purified folate-binding protein from cow's milk was immobilized on monodisperse polymer particles (Dynospheres) activated by rho-toluenesulfonyl chloride. Leakage from the spheres was less than 0.1%, and the binding properties were similar to those of the soluble protein with regard to dissociation, pH optimum for binding pteroylglutamic acid, and specificity for binding various folate derivatives. We used the immobilized folate-binding protein as binding protein in an isotope-dilution assay for quantifying folate in erythrocytes. The detection limit was 50 nmol/L and the CV over a six-month period was 2.3% (means = 1.25 mumol/L, n = 15). The reference interval, for folate measured in erythrocytes of 43 blood donors, was 0.4-1.5 mumol/L.

  2. The Failure of Purified T-2 Mycotoxin to Produce Hemorrhaging in Dairy Cattle

    PubMed Central

    Weaver, G. A.; Kurtz, H. J.; Mirocha, C. J.; Bates, F. Y.; Behrens, J. C.; Robison, T. S.; Swanson, S. P.

    1980-01-01

    A Holstein cow was intubated with 182 mg of 97% pure T-2 toxin (0.44 mg/kg of body weight) for 15 days. A dairy ration containing 50 mg/kg (50 ppm) of T-2 toxin was refused. A calf, born four days after onset of maternal treatment, was intubated with 26.2 mg of purified T-2 toxin (0.6 mg/kg of body weight) for seven consecutive days and then on alternate days for a total of 16 days. The calf was severely affected clinically by the T-2 toxin. The T-2 toxin failed to cause bovine hemorrhagic syndrome in either animal. Unspecific gastrointestinal lesions were noted in the cow but none were detected in the calf. In the calf, severe depression, hindquarter ataxia, knuckling of the rear feet, listlessness and anorexia were caused by the T-2 toxin. PMID:7427850

  3. Antifungal activity of violacein purified from a novel strain of Chromobacterium sp. NIIST (MTCC 5522).

    PubMed

    Sasidharan, Anju; Sasidharan, Nishanth Kumar; Amma, Dileepkumar Bhaskaran Nair Saraswathy; Vasu, Radhakrishnan Kokkuvayil; Nataraja, Anupama Vijaya; Bhaskaran, Krishnakumar

    2015-10-01

    A novel strain of Chromobacterium sp. NIIST (MTCC 5522) producing high level of purple blue bioactive compound violacein was isolated from clay mine acidic sediment. During 24 h aerobic incubation in modified Luria Bertani medium, around 0.6 g crude violacein was produced per gram of dry weight biomass. An inexpensive method for preparing crystalline, pure violacein from crude pigment was developed (12.8 mg violacein/L) and the pure compound was characterized by different spectrometric methods. The violacein prepared was found effective against a number of plant and human pathogenic fungi and yeast species such as Cryptococcus gastricus, Trichophyton rubrum, Fusarium oxysporum, Rhizoctonia solani, Aspergillus flavus, Penicillium expansum, and Candida albicans. The best activity was recorded against Trichophyton rubrum (2 -g/ml), a human pathogen responsible for causing athlete-s foot infection. This is the first report of antifungal activity of purified violacein against pathogenic fungi and yeast.

  4. A Method of Effective Quarry Water Purifying Using Artificial Filtering Arrays

    NASA Astrophysics Data System (ADS)

    Tyulenev, M.; Garina, E.; Khoreshok, A.; Litvin, O.; Litvin, Y.; Maliukhina, E.

    2017-01-01

    The development of open pit mining in the large coal basins of Russia and other countries increases their negative impact on the environment. Along with the damage of land and air pollution by dust and combustion gases of blasting, coal pits have a significant negative impact on water resources. Polluted quarry water worsens the ecological situation on a much larger area than covered by air pollution and land damage. This significantly worsens the conditions of people living in cities and towns located near the coal pits, and complicates the subsequent restoration of the environment, irreversibly destroying the nature. Therefore, the research of quarry wastewater purifying is becoming an important mater for scholars of technical colleges and universities in the regions with developing open-pit mining. This paper describes the method of determining the basic parameters of the artificial filtering arrays formed on coal pits of Kuzbass (Western Siberia, Russia), and gives recommendations on its application.

  5. A study of cell electrophoresis as a means of purifying growth hormone secreting cells

    NASA Technical Reports Server (NTRS)

    Plank, Lindsay D.; Hymer, W. C.; Kunze, M. Elaine; Marks, Gary M.; Lanham, J. Wayne

    1983-01-01

    Growth hormone secreting cells of the rat anterior pituitary are heavily laden with granules of growth hormone and can be partialy purified on the basis of their resulting high density. Two methods of preparative cell electrophoresis were investigated as methods of enhancing the purification of growth hormone producing cells: density gradient electrophoresis and continuous flow electrophoresis. Both methods provided a two- to four-fold enrichment in growth hormone production per cell relative to that achieved by previous methods. Measurements of electrophoretic mobilities by two analytical methods, microscopic electrophoresis and laser-tracking electrophoresis, revealed very little distinction between unpurified anterior pituitary cell suspensions and somatotroph-enriched cell suspensions. Predictions calculated on the basis of analytical electrophoretic data are consistent with the hypothesis that sedimentation plays a significant role in both types of preparative electrophoresis and the electrophoretic mobility of the growth hormone secreting subpopulation of cells remains unknown.

  6. Characterization of phage receptors in Streptococcus thermophilus using purified cell walls obtained by a simple protocol.

    PubMed

    Quiberoni, A; Stiefel, J I; Reinheimer, J A

    2000-12-01

    A simple protocol was designed and applied to obtain Streptococcus thermophilus purified cell walls. To identify the structures involved in phage adsorption, the cell walls of two Strep. thermophilus strains were treated with sodium dodecyl sulphate and proteinase K. These treatments did not reduce the adsorption of phages CYM and 0BJ to the cell walls of Strep. thermophilus YSD10 and Strep. thermophilus BJ15, respectively. However, phage binding was reduced when the cell envelopes were treated with mutanolysin or trichloroacetic acid 5%, suggesting that the phage receptor component is part of the peptidoglycan or a polymer closely linked to it. The ability of several saccharides to inactivate both phages was also assayed. These phage inhibition experiments suggested that the phage CYM adsorbed to a component involving glucosamine and rhamnose, while glucosamine and ribose interfered with the adsorption of phage 0BJ.

  7. Structure characteristics of a water-soluble polysaccharide purified from dragon fruit (Hylocereus undatus) pulp.

    PubMed

    Xu, Lishan; Zhang, Yaojie; Wang, Lizhi

    2016-08-01

    Dragon fruit is a tropical fruit with good taste. It can bring health benefits to human body. As one of the major bioactive components in this fruit, the polysaccharides might contribute to the health benefits. However, the precise structure information remains unknown. A leading polysaccharide of dragon fruit pulp, DFPP, was purified and identified by NMR and GC-MS. →4-β-d-GlcpA-1→, →6-β-d-Galp-1→ and →4-α-l-Rhap-1→ constituted the backbone and α-l-Araf-1→5-α-l-Araf-1→ formed the branch chain. The precise structure was putatively identified as below. The molecular weight was 2.2×10(3)kDa. The structure information of polysaccharides will be helpful to understand this fruit.

  8. Highly purified hexachlorobenzene induces cytochrome P4501A in primary cultures of chicken embryo hepatocytes

    SciTech Connect

    Mundy, Lukas J.; Jones, Stephanie P.; Crump, Doug; Herve, Jessica C.; Konstantinov, Alex; Utley, Fiona; Potter, David; Kennedy, Sean W.

    2010-11-01

    Some uncertainty exists regarding the purity of hexachlorobenzene (HCB) used in past toxicity studies. It has been suggested that reported toxic and biochemical effects initially attributed to HCB exposure may have actually been elicited by contamination of HCB by polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs). Herein, primary cultures of chicken embryo hepatocytes (CEH) were used to compare the potencies of two lots of reagent-grade hexachlorobenzene (HCB-old [HCB-O] and HCB-new [HCB-N]), highly purified HCB (HCB-P) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) as inducers of ethoxyresorufin O-deethylase (EROD) activity, cytochrome P4501A4 (CYP1A4) messenger ribonucleic acid (mRNA) and CYP1A5 mRNA. The study also compared the EROD- and CYP1A4/5 mRNA-inducing potencies of HCB to the potencies of two mono-ortho substituted polychlorinated biphenyls (PCBs), 2,3,3',4,4'-pentachlorobiphenyl (PCB 105) and 2,3'4,4',5-pentachlorobiphenyl (PCB 118). HCB-O, HCB-N and HCB-P all induced EROD activity and up-regulated CYP1A4 and CYP1A5 mRNAs. Induction was not caused by contamination of HCB with PCDDs or PCDFs. Based upon a comparison of the EC{sub 50} and EC{sub threshold} values for EROD and CYP1A4/5 mRNA concentration-response curves, the potency of HCB relative to the potency of TCDD was 0.0001, and was similar to that of PCB 105 and PCB 118. The maximal EROD activity and CYP1A4/5 mRNA expression differed greatly between HCB and TCDD, and may contribute to an overestimation of the ReP value calculated for highly purified HCB.

  9. Pirenzepine binding to membrane-bound, solubilized and purified muscarinic receptor subtypes

    SciTech Connect

    Baumgold, J.

    1986-05-01

    Muscarinic receptors were purified to near-homogeneity from bovine cortex, an area rich in the putative M1 subtype, and from bovine pons/medulla, an area rich in the putative M2 subtype. In both cases, the receptors were solubilized in digitonin and purified over an affinity column. Both the cortical and pons/medulla preparations yielded receptor proteins of 70,000 daltons. Pirenzepine binding was deduced from its competition with /sup 3/H-N-methyl scopolamine. The binding of pirenzepine to membrane-bound receptors from cortex was best described by a two site model, with approximately half the sites having a Ki of 6.4 x 10/sup -9/ M and the remaining sites having a Ki of 3.5 x 10/sup -7/ M. Membrane-bound receptors from pons/medulla bound pirenzepine according to a one-site model with a Ki of 1.1 x 10/sup -7/ M. After solubilization the two-site binding of cortical receptors became a one-site binding, Ki = 1.1 x 10/sup -7/M. This value was still five-fold lower than that of soluble receptors from pons/medulla. After purification however the affinity of pirenzepine for the pons/medulla receptor increased so that the two putative subtypes bound pirenzepine with approximately the same affinity. These findings suggest that the different pirenzepine binding characteristics used to define muscarinic receptor subtypes are not inherent in the receptor protein itself but may be due to coupling factors associated with the receptor.

  10. Purified human SUV3p exhibits multiple-substrate unwinding activity upon conformational change.

    PubMed

    Shu, Zhanyong; Vijayakumar, Sangeetha; Chen, Chi-Fen; Chen, Phang-Lang; Lee, Wen-Hwa

    2004-04-27

    Suv3 of Saccharomyces cerevisiae has been classified as a mitochondrial RNA helicase. However, the helicase domain in both yeast and human SUV3 varies considerably from the typical RNA helicase motifs. To investigate its enzymatic activities, a homogeneously purified preparation of SUV3 is required. Expression of a processed form of human SUV3 carrying an N-terminal deletion of 46 amino acids (SUV3DeltaN46) in a yeast suv3 null mutant, which otherwise fails to grow in a nonfermentable carbon source and forms petite colonies in glucose medium, rescues the null phenotype. Through a five-step chromatographic procedure, an 83 kDa SUV3DeltaN46 protein (SUV3-83) and a partially degraded 70 kDa product (SUV3-70) containing amino acids 68-685 were purified to homogeneity. Single- or double-stranded DNA and RNA stimulated ATPase activity of both proteins. SUV3-70, which retains core catalytic domains, can bind and unwind multiple duplex substrates of RNA and DNA with a 5'-3' directionality over a wide range of pH, while SUV3-83 has helicase activity at only acidic pH. ATP, but not nonhydrolyzable ATP, is essential for the unwinding activity, suggesting the requirement of the energy derived from ATP hydrolysis. Consistent with this notion, suv3 mutants containing alanine (A) or arginine (R) substitutions at the conserved lysine residue in the ATP binding site (K213) lost ATPase activity and also failed to unwind the substrates. Importantly, circular dichroism (CD) spectral analysis showed that SUV3-83, at pH 5.0, adopts a conformation similar to that of SUV3-70, suggesting a conformational change in SUV3-83 is required for its helicase activity. The physiological relevance of the multiple-substrate helicase activity of human SUV3 is discussed.

  11. Studies on the hyaluronate binding properties of newly synthesized proteoglycans purified from articular chondrocyte cultures

    SciTech Connect

    Sandy, J.D.; Plaas, A.H.

    1989-06-01

    Primary cultures of rabbit articular chondrocytes have been maintained for 10 days and labeled with (35S)sulfate, (3H)leucine, and (35S)cysteine in pulse-chase protocols to study the structure and hyaluronate binding properties of newly synthesized proteoglycan monomers. Radiolabeled monomers were purified from medium and cell-layer fractions by dissociative CsCl gradient centrifugation with bovine carrier monomer, and analyzed for hyaluronate binding affinity on Sepharose CL-2B in 0.5 M Na acetate, 0.1% Triton X-100, pH 6.8. Detergent was necessary to prevent self-association of newly synthesized monomers during chromatography. Monomers secreted during a 30-min pulse labeling with (35S)sulfate had a low affinity relative to carrier. Those molecules released into the medium during the first 12 h of chase remained in the low affinity form whereas those retained by the cell layer rapidly acquired high affinity. In cultures where more than 90% of the preformed cell-layer proteoglycan was removed by hyaluronidase digestion before radiolabeling the newly synthesized low affinity monomers also rapidly acquired high affinity if retained in the cell layer. Cultures labeled with amino acid precursors were used to establish the purity of monomer preparations and to isolate core proteins for study. Leucine- or cysteine-labeled core proteins derived from either low or high affinity monomer preparations migrated as a single major species on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with electrophoretic mobility very similar to that of core protein derived from extracted proteoglycan monomer. Purified low affinity monomers were converted to the high affinity form by treatment at pH 8.6; however, this change was prevented by guanidinium-HCl at concentrations above 0.8 M.

  12. Physicochemical and Biological Characterization of Fucoidan from Fucus vesiculosus Purified by Dye Affinity Chromatography

    PubMed Central

    Zayed, Ahmed; Muffler, Kai; Hahn, Thomas; Rupp, Steffen; Finkelmeier, Doris; Burger-Kentischer, Anke; Ulber, Roland

    2016-01-01

    A comparative study concerning the physicochemical, monomeric composition and biological characters among different fucoidan fractions is presented. Common purification techniques for fucoidan usually involve many steps. During these steps, the important structural features might be affected and consequently alter its biological activities. Three purified fractions were derived from Fucus vesiculosus water extract which, afterwards, were purified by a recently-developed dye affinity chromatography protocol. This protocol is based on dye-sulfated polysaccharide interactions. The first two fractions were obtained from crude precipitated fucoidan at different pH values of the adsorption phase: pH 1 and 6. This procedure resulted in fucoidan_1 and 6 fractions. The other, third, fraction: fucoidan_M, however, was obtained from a buffered crude extract at pH 1, eliminating the ethanol precipitation step. All of the three fractions were then further evaluated. Results revealed that fucoidan_M showed the highest sulfur content (S%), 12.11%, with the lowest average molecular weight, 48 kDa. Fucose, galactose, and uronic acid/glucose dimers were detected in all fractions, although, xylose was only detected in fucoidan_1 and 6. In a concentration of 10 µg·mL−1, Fucoidan_6 showed the highest heparin-like anticoagulant activity and could prolong the APTT and TT significantly to 66.03 ± 2.93 and 75.36 ± 1.37 s, respectively. In addition, fucoidan_M demonstrated the highest potency against HSV-1 with an IC50 of 2.41 µg·mL−1. The technique proved to be a candidate for fucoidan purifaction from its crude extract removing the precipitation step from common purification protocols and produced different fucoidan qualities resulted from the different incubation conditions with the immobilized thiazine toluidine blue O dye. PMID:27092514

  13. Use of recombinant purified protein derivative (PPD) antigens as specific skin test for tuberculosis

    PubMed Central

    Stavri, Henriette; Bucurenci, Nadia; Ulea, Irina; Costache, Adriana; Popa, Loredana; Popa, Mircea Ioan

    2012-01-01

    Background & objectives: Purified protein derivative (PPD) is currently the only available skin test reagent used worldwide for the diagnosis of tuberculosis (TB). The aim of this study was to develop a Mycobacterium tuberculosis specific skin test reagent, without false positive results due to Bacillus Calmette-Guerin (BCG) vaccination using recombinant antigens. Methods: Proteins in PPD IC-65 were analyzed by tandem mass spectrometry and compared to proteins in M. tuberculosis culture filtrate; 54 proteins were found in common. Top candidates MPT64, ESAT 6, and CFP 10 were overexpressed in Escherichia coli expression strains and purified as recombinant proteins. To formulate optimal immunodiagnostic PPD cocktails, the antigens were evaluated by skin testing guinea pigs sensitized with M. tuberculosis H37Rv and BCG. Results: For single antigens and a cocktail mixture of these antigens, best results were obtained using 3 μg/0.1 ml, equivalent to 105 TU (tuberculin units). Each animal was simultaneously tested with PPD IC-65, 2 TU/0.1 ml, as reference. Reactivity of the multi-antigen cocktail was greater than that of any single antigen. The skin test results were between 34.3 and 76.6 per cent the level of reactivity compared to that of the reference when single antigens were tested and 124 per cent the level of reactivity compared to the reference for the multi-antigen cocktail. Interpretation & conclusions: Our results showed that this specific cocktail could represent a potential candidate for a new skin diagnostic test for TB. PMID:23287127

  14. Antioxidant and ACE Inhibitory Bioactive Peptides Purified from Egg Yolk Proteins

    PubMed Central

    Yousr, Marwa; Howell, Nazlin

    2015-01-01

    Protein by-products from the extraction of lecithin from egg yolk can be converted into value-added products, such as bioactive hydrolysates and peptides that have potential health enhancing antioxidant, and antihypertensive properties. In this study, the antioxidant and angiotensin converting enzyme (ACE) inhibitory activities of peptides isolated and purified from egg yolk protein were investigated. Defatted egg yolk was hydrolyzed using pepsin and pancreatin and sequentially fractionated by ultrafiltration, followed by gel filtration to produce egg yolk gel filtration fractions (EYGF). Of these, two fractions, EYGF-23 and EYGF-33, effectively inhibited the peroxides and thiobarbituric acid reactive substance (TBARS) in an oxidizing linoleic acid model system. The antioxidant mechanism involved superoxide anion and hydroxyl radicals scavenging and ferrous chelation. The presence of hydrophobic amino acids such as tyrosine (Y) and tryptophan (W), in sequences identified by LC-MS as WYGPD (EYGF-23) and KLSDW (EYGF-33), contributed to the antioxidant activity and were not significantly different from the synthetic BHA antioxidant. A third fraction (EYGF-56) was also purified from egg yolk protein by gel filtration and exhibited high ACE inhibitory activity (69%) and IC50 value (3.35 mg/mL). The SDNRNQGY peptide (10 mg/mL) had ACE inhibitory activity, which was not significantly different from that of the positive control captopril (0.5 mg/mL). In addition, YPSPV in (EYGF-33) (10 mg/mL) had higher ACE inhibitory activity compared with captopril. These findings indicated a substantial potential for producing valuable peptides with antioxidant and ACE inhibitory activity from egg yolk. PMID:26690134

  15. Measuring the effect of photocatalytic purifiers on indoor air hydrocarbons and carbonyl pollutants.

    PubMed

    Disdier, Jean; Pichat, Pierre; Mas, Denis

    2005-01-01

    Laboratory tests of photocatalytic air purifiers are usually performed with a single pollutant, in the parts per million by volume domain and at airflow rates < or =0.1 m3/hr. Clearly, it is necessary to probe photocatalytic materials and apparatuses under real conditions or conditions closely mimicking reality. Photocatalytic prototypes were placed in an ordinary room. To collect hydrocarbons over a shorter period (15 min) than with adsorbent-containing cartridges, solid-phase microextraction (SPME) was used. Typically, concentrations in substituted benzene hydrocarbons and tetrachloroethene were decreased to 20-35% of initial values; toluene and m- + p-xylene concentrations dropped to 2-6 parts per billion by volume, and o-xylene and benzene concentrations were still lower. In the absence of appropriate, commercialized SPME fibers, carbonyl compounds (both formed and destroyed by photocatalysis) were extracted using cartridges containing 2,4- dinitrophenylhydrazine-coated silica. The concentration ranges (in parts per billion by volume) were shifted to higher values in treated air: from 9-15.5 to 12.5-18 for methanal, from 1.5-3 to 8-11.5 for ethanal, and from 4.5-19 to 8-26.5 for propanone with the prototype used; these unprecedented results do not exclude using photocatalysis to treat air, but they illustrate that improvement is needed. Because these tests are time-consuming, preliminary tests are useful; results obtained with a 225-L closed-loop, airtight, photocatalytic reactor with an external turbine enabling the ambient air inside the reactor to be circulated through the purifier device at 15-450 m3/hr flow rates are reported.

  16. Studies on the bioactivity of radioiodinated highly purified bovine thyrotropin: analytical polyacrylamide gel electrophoresis

    SciTech Connect

    Takai, N.A.; Filetti, S.; Rapoport, B.

    1981-01-01

    Highly purified bovine TSH (stored in solution at -70 C) was radioiodinated by the stoichiometric chloroamine-T method. The iodinated material ws subjected to analytical polyacrylamide disc gel electrophoresis. TSH was eluted from gel slices (1 mm width) and was analyzed for radioactivity and bioactivity. The latter was determined using the cultured thyroid cell cAMP response assay. Radioactivity in the TSH preparation migrated separately from bioactivity, but concordant with the protein bands observed in gels run in parallel. Further studies performed on bovine TSH purified in our laboratory, as well as on a different TSH preparation of exceptionally high potency (both stored as lyophilized powder) revealed a different pattern, with TSH bioactivity and radioactivity eluting concurrently. Iodination of TSH did not alter its electrophoretic migration on disc gel electrophoresis. In all preparations polymorphism of TSH bioactivity was observed, with at least four separate protein bands containing TSH bioactivity being present in our preparation. The relationship between the degree of iodination and retention of TSH bioactivity was examined. Incorporation of /sup 125/I into TSH was greatly different at two different concentrations of chloramine-T. Despite this, however, the progressive loss of TSH bioactivity was similar at both concentrations, indicating that incorporation of iodine into the TSH molecule is not itself responsible for the decrease in bioactivity. These studies indicate variability among different TSH preparations in terms of their retention of bioactivity. Significant loss of TSH bioactivity appears to occur during storage in solution. The damage to the biological activity of TSH during the iodination procedure is more likely related to the oxidation process than to the incorporation of iodine.

  17. Superovulation of goats with purified pFSH supplemented with defined amounts of pLH.

    PubMed

    Nowshari, M A; Backers, J F; Holtz, W

    1995-03-01

    The superovulatory response of goats treated with purified pFSH supplemented with 30, 40 or 50% pLH was compared. Sixty-four Boer goat does were synchronized by progestagen-containing ear implant, randomly allotted to 3 groups and, beginning 2 d before implant removal, treated with purified pFSH supplemented with 30, 40 or 50% pLH. Each animal received 16 Armour Units of pFSH administered in 6 descending doses at 12-h intervals. Along with the last 2 injections, the does received 5 mg PGF(2alpha). Embryos were flushed either surgically or after slaughter on Day 5 or 6 after the last day of standing estrus. The percentage of animals responding to treatment was not different among groups treated with pFSH supplemented with 30, 40 or 50% pLH (76, 71 and 63%, respectively). The corresponding data for number of ovulations was 11.3 +/- 1.6, 16.3 +/- 1.8 and 16.4 +/- 2.6, for number of ova and embryos recovered 8.1 +/- 1.9, 12.0 +/- 1.5 and 13.5 +/- 2.9 and for number of transferable embryos 6.6 +/- 1.9, 9.1 +/- 1.5 and 7.1 +/- 2.1 (x +/- SEM). Results confirm the earlier finding of a good response of goats to pFSH preparations with a high FSH:LH ratio, and, although group differences were statistically nonsignificant (P > 0.05), they suggest that supplementation with approximately 40% pLH may be close to the optimum.

  18. Antioxidant and ACE Inhibitory Bioactive Peptides Purified from Egg Yolk Proteins.

    PubMed

    Yousr, Marwa; Howell, Nazlin

    2015-12-07

    Protein by-products from the extraction of lecithin from egg yolk can be converted into value-added products, such as bioactive hydrolysates and peptides that have potential health enhancing antioxidant, and antihypertensive properties. In this study, the antioxidant and angiotensin converting enzyme (ACE) inhibitory activities of peptides isolated and purified from egg yolk protein were investigated. Defatted egg yolk was hydrolyzed using pepsin and pancreatin and sequentially fractionated by ultrafiltration, followed by gel filtration to produce egg yolk gel filtration fractions (EYGF). Of these, two fractions, EYGF-23 and EYGF-33, effectively inhibited the peroxides and thiobarbituric acid reactive substance (TBARS) in an oxidizing linoleic acid model system. The antioxidant mechanism involved superoxide anion and hydroxyl radicals scavenging and ferrous chelation. The presence of hydrophobic amino acids such as tyrosine (Y) and tryptophan (W), in sequences identified by LC-MS as WYGPD (EYGF-23) and KLSDW (EYGF-33), contributed to the antioxidant activity and were not significantly different from the synthetic BHA antioxidant. A third fraction (EYGF-56) was also purified from egg yolk protein by gel filtration and exhibited high ACE inhibitory activity (69%) and IC50 value (3.35 mg/mL). The SDNRNQGY peptide (10 mg/mL) had ACE inhibitory activity, which was not significantly different from that of the positive control captopril (0.5 mg/mL). In addition, YPSPV in (EYGF-33) (10 mg/mL) had higher ACE inhibitory activity compared with captopril. These findings indicated a substantial potential for producing valuable peptides with antioxidant and ACE inhibitory activity from egg yolk.

  19. Regenerated silica gel as stationary phase on vacuum column chromatography to purify temulawak's extracts

    NASA Astrophysics Data System (ADS)

    Cahyono, Bambang; Maduwu, Ratna Dewi; Widayat, Suzery, Meiny

    2015-12-01

    Commercial silica gel only used once by many researchers and affected high cost for purification process, also less support the green chemistry program. This research focused in regeneration silica gel that used purification of temulawak's extracts (Curcuma xanthorrhiza Roxb) by vacuum column chromatography. Sample extracts (contains 10.1195±0.5971% of curcuminoids) was purified by vacuum column chromatography (pressure: 45 kPa, column: 100mm on length and 16mm on diameter). Ethanol 96% and acetone were compared as eluent. The amount of solvent and yield of curcuminoids used as indicator purification. The silica gel was regenerated with heating in 600°C for 8 hours The silica gels were analyzed by IR spectroscopy and X-ray diffraction. Furthermore, regenerated silica gel was used as the stationary phase in vacuum column chromatography under the same conditions with the previous purification. All the purification experiments were performed in three repetitions. Based on regression equation, y=0.132x+0.0011 (r2=0.9997) the yield of curcuminoids on purified products using ethanol as the eluent was improved 4.26% (to 14.3724±0.5749%) and by acetone was improved 3,03% (to 13.1450 ±0.6318%). The IR spectrum of both silica gel showed the same vibration profile and also there were three crystallinity peaks missing on its X-ray diffraction. Regenerated silica gel has the same performance with new silica gel in purification of temulawak's extract: by ethanol has increased 4.08% (14.1947±0.7415%) and 2.93% (13.0447±0.4822) by acetone. In addition, all purification products showed similar TLC profiles. Purification using regenerated silica gel as the adsorbent on vacuum column chromatography has exactly same potential with the new silica gel.

  20. Nucleic acid binding proteins in highly purified Creutzfeldt-Jakob disease preparations.

    PubMed Central

    Sklaviadis, T; Akowitz, A; Manuelidis, E E; Manuelidis, L

    1993-01-01

    The nature of the infectious agent causing human Creutzfeldt-Jakob disease (CJD), a slowly progressive dementia, is controversial. As in scrapie, no agent-specific proteins or nucleic acids have been identified. However, biological features of exponential replication and agent strain variation, as well as physical size and density data, are most consistent with a viral structure--i.e., a nucleic acid-protein complex. It is often assumed that nuclease treatment, which does not reduce infectious titer, leaves no nucleic acids of > 50 bp. However, nucleic acids of 500-6000 bp can be extracted from highly purified infectious complexes with a mass of approximately 1.5 x 10(7) daltons. It was therefore germane to search for nucleic acid binding proteins that might protect an agent genome. We here use Northwestern blotting to show that there are low levels of nonhistone nucleic acid binding proteins in highly purified infectious 120S gradient fractions. Several nucleic acid binding proteins were clearly host encoded, whereas others were apparent only in CJD, but not in parallel preparations from uninfected brain. Small amounts of residual host Gp34 (prion protein) did not bind any 32P-labeled nucleic acid probes. Most of the minor "CJD-specific" proteins had an acidic pI, a characteristic of many viral core proteins. Such proteins deserve further study, as they probably contribute to unique properties of resistance described for these agents. It remains to be seen if any of these proteins are agent encoded. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 PMID:8516321

  1. Hemizygosity Enhances Purifying Selection: Lack of Fast-Z Evolution in Two Satyrine Butterflies

    PubMed Central

    Rousselle, Marjolaine; Faivre, Nicolas; Ballenghien, Marion; Galtier, Nicolas; Nabholz, Benoit

    2016-01-01

    The fixation probability of a recessive beneficial mutation is increased on the X or Z chromosome, relative to autosomes, because recessive alleles carried by X or Z are exposed to selection in the heterogametic sex. This leads to an increased dN/dS ratio on sex chromosomes relative to autosomes, a pattern called the “fast-X” or “fast-Z” effect. Besides positive selection, the strength of genetic drift and the efficacy of purifying selection, which affect the rate of molecular evolution, might differ between sex chromosomes and autosomes. Disentangling the complex effects of these distinct forces requires the genome-wide analysis of polymorphism, divergence and gene expression data in a variety of taxa. Here we study the influence of hemizygosity of the Z chromosome in Maniola jurtina and Pyronia tithonus, two species of butterflies (Lepidoptera, Nymphalidae, Satyrinae). Using transcriptome data, we compare the strength of positive and negative selection between Z and autosomes accounting for sex-specific gene expression. We show that M. jurtina and P. tithonus do not experience a faster, but rather a slightly slower evolutionary rate on the Z than on autosomes. Our analysis failed to detect a significant difference in adaptive evolutionary rate between Z and autosomes, but comparison of male-biased, unbiased and female-biased Z-linked genes revealed an increased efficacy of purifying selection against recessive deleterious mutations in female-biased Z-linked genes. This probably contributes to the lack of fast-Z evolution of satyrines. We suggest that the effect of hemizygosity on the fate of recessive deleterious mutations should be taken into account when interpreting patterns of molecular evolution in sex chromosomes vs. autosomes. PMID:27590089

  2. Regenerated silica gel as stationary phase on vacuum column chromatography to purify temulawak’s extracts

    SciTech Connect

    Cahyono, Bambang; Maduwu, Ratna Dewi; Widayat,; Suzery, Meiny

    2015-12-29

    Commercial silica gel only used once by many researchers and affected high cost for purification process, also less support the green chemistry program. This research focused in regeneration silica gel that used purification of temulawak’s extracts (Curcuma xanthorrhiza Roxb) by vacuum column chromatography. Sample extracts (contains 10.1195±0.5971% of curcuminoids) was purified by vacuum column chromatography (pressure: 45 kPa, column: 100mm on length and 16mm on diameter). Ethanol 96% and acetone were compared as eluent. The amount of solvent and yield of curcuminoids used as indicator purification. The silica gel was regenerated with heating in 600°C for 8 hours The silica gels were analyzed by IR spectroscopy and X-ray diffraction. Furthermore, regenerated silica gel was used as the stationary phase in vacuum column chromatography under the same conditions with the previous purification. All the purification experiments were performed in three repetitions. Based on regression equation, y=0.132x+0.0011 (r{sup 2}=0.9997) the yield of curcuminoids on purified products using ethanol as the eluent was improved 4.26% (to 14.3724±0.5749%) and by acetone was improved 3,03% (to 13.1450 ±0.6318%). The IR spectrum of both silica gel showed the same vibration profile and also there were three crystallinity peaks missing on its X-ray diffraction. Regenerated silica gel has the same performance with new silica gel in purification of temulawak’s extract: by ethanol has increased 4.08% (14.1947±0.7415%) and 2.93% (13.0447±0.4822) by acetone. In addition, all purification products showed similar TLC profiles. Purification using regenerated silica gel as the adsorbent on vacuum column chromatography has exactly same potential with the new silica gel.

  3. Development program to recycle and purify plutonium-238 oxide fuel from scrap

    NASA Astrophysics Data System (ADS)

    Schulte, Louis D.; Silver, Gary L.; Avens, Larry R.; Jarvinen, Gordon D.; Espinoza, Jacob; Foltyn, Elizabeth M.; Rinehart, Gary H.

    1997-01-01

    Nuclear Materials Technology (NMT) Division of Los Alamos National Laboratory (LANL) has initiated a development program to recover & purify plutonium-238 oxide from impure sources. A glove box line has been designed and a process flowsheet developed to perform this task on a large scale. Our initial effort has focused on purification of 238PuO2 fuel that fails to meet General Purpose Heat Source (GPHS) specifications because of impurities. The most notable non-actinide impurity was silicon, but aluminum, chromium, iron and nickel were also near or in excess of limits specified by GPHS fuel powder specifications. 234U was by far the largest actinide impurity observed in the feed material because it is the daughter product of 238Pu by alpha decay. An aqueous method based on nitric acid was selected for purification of the 238PuO2 fuel. All aqueous processing used high purity reagents, and was performed in PTFE apparatus to minimize introduction of new contaminants. Impure 238PuO2 was finely milled, then dissolved in refluxing HNO3/HF and the solution filtered. The dissolved 238Pu was adjusted to the trivalent state by an excess of reducing reagents to compensate for radiolytic effects, precipitated as plutonium(III) oxalate, and recovered by filtration. The plutonium(III) oxalate was subsequently calcined to convert the plutonium to the oxide. Decontamination factors for silicon, phosphorus and uranium were excellent. Decontamination factors for aluminum, chromium, iron and nickel were very good. The purity of the 238PuO2 recovered from this operation was significantly better than specifications. Efforts continue to develop the capability for efficient, safe, cost-effective, and environmentally acceptable methods to recover and purify 238PuO2 fuel in a glove box environment. Plutonium-238 materials targeted for recovery includes impure oxide and scrap items that are lean in 238Pu values.

  4. Physicochemical and Biological Characterization of Fucoidan from Fucus vesiculosus Purified by Dye Affinity Chromatography.

    PubMed

    Zayed, Ahmed; Muffler, Kai; Hahn, Thomas; Rupp, Steffen; Finkelmeier, Doris; Burger-Kentischer, Anke; Ulber, Roland

    2016-04-15

    A comparative study concerning the physicochemical, monomeric composition and biological characters among different fucoidan fractions is presented. Common purification techniques for fucoidan usually involve many steps. During these steps, the important structural features might be affected and consequently alter its biological activities. Three purified fractions were derived from Fucus vesiculosus water extract which, afterwards, were purified by a recently-developed dye affinity chromatography protocol. This protocol is based on dye-sulfated polysaccharide interactions. The first two fractions were obtained from crude precipitated fucoidan at different pH values of the adsorption phase: pH 1 and 6. This procedure resulted in fucoidan_1 and 6 fractions. The other, third, fraction: fucoidan_M, however, was obtained from a buffered crude extract at pH 1, eliminating the ethanol precipitation step. All of the three fractions were then further evaluated. Results revealed that fucoidan_M showed the highest sulfur content (S%), 12.11%, with the lowest average molecular weight, 48 kDa. Fucose, galactose, and uronic acid/glucose dimers were detected in all fractions, although, xylose was only detected in fucoidan_1 and 6. In a concentration of 10 µg·mL(-1), Fucoidan_6 showed the highest heparin-like anticoagulant activity and could prolong the APTT and TT significantly to 66.03 ± 2.93 and 75.36 ± 1.37 s, respectively. In addition, fucoidan_M demonstrated the highest potency against HSV-1 with an IC50 of 2.41 µg·mL(-1). The technique proved to be a candidate for fucoidan purifaction from its crude extract removing the precipitation step from common purification protocols and produced different fucoidan qualities resulted from the different incubation conditions with the immobilized thiazine toluidine blue O dye.

  5. [A primary clinical trial of genital warts treated with domestic highly purified podophyllotoxin].

    PubMed

    Wang, B; Wang, B; Shao, Y

    1994-04-01

    Genital warts (condyloma accunimata) are a kind of sexually transmitted disease (STD) caused by human papillomavirus (HPV). In this article, we report 75 cases of genital warts treated with highly purified podophyllotoxin. All volunteers were selected in our clinic and diagnosed according to typical skin lesions and positive 0.5% acetic acid test. Of the 74 cases 48 were male and 26 were female, with a mean age of 28.4 years (21-54). The average duration of disease was 4.4 months (2 weeks-3 years). All lesions were treated with 0.5% podophyllotoxin-ethanal solution topically 2 times per day for 3 successive days. The treatment was repeated if any warts persisted, but not more than 3 therapies in total were applied. The results showed that 41/48 (85.4%) of male and 23/26 (88.5%) of female patients were cured, the overall cure rate was 86.5% and the effectiveness rate (more than 50% lesions disappeared) was 96.15%. During another 3 months of follow-up, 9 male and 4 female patients (17.6% in total) experienced relapsed warts; these were cured with another treatment. Side effects including slight local irritation and erythema or superficial erosions were common, but 3 male and 2 female patients complained of sever pain and local swelling. All side effects were tolerable. It is concluded that our domestically produced highly purified podophyllotoxin is an effective, comparatively safe drug for the treatment of genital warts. It is also very simple to apply, and can be used for self-treatment.

  6. Transfection Studies with Colloidal Systems Containing Highly Purified Bipolar Tetraether Lipids from Sulfolobus acidocaldarius

    PubMed Central

    Pinnapireddy, Shashank Reddy; Baghdan, Elias; Jedelská, Jarmila

    2017-01-01

    Lipid vectors are commonly used to facilitate the transfer of nucleic acids into mammalian cells. In this study, two fractions of tetraether lipids from the archaea Sulfolobus acidocaldarius were extracted and purified using different methods. The purified lipid fractions polar lipid fraction E (PLFE) and hydrolysed glycerol-dialkyl-nonitol tetraether (hGDNT) differ in their structures, charge, size, and miscibility from conventional lipids. Liposomes were prepared by mixing tetraether lipids with cholesterol (CH) and 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) resulting in stable vectors for gene delivery. Lipoplexes were prepared by complexation of liposomes with a luciferase expressing plasmid (pCMV-luc) at certain nitrogen-to-phosphorus (N/P) ratios and optimised for the transient transfection of ovarian adenocarcinoma cells (SK-OV-3). Complexation efficacy was investigated by gel-red fluorescence assay. Biophysical properties, like size, surface charge, and morphology, were investigated by differential light scattering (DLS), atomic force microscopy (AFM), and scanning electron microscopy (Cryo-SEM), respectively, revealing structural differences between liposomes and lipoplexes. A range of stable transfecting agents containing tetraether lipids were obtained by incorporating 5 mol% of tetraether lipids. Lipoplexes showed a decrease in free gel-red with increasing N/P ratios indicating efficient incorporation of plasmid DNA (pDNA) and remarkable stability. Transfection experiments of the lipoplexes revealed successful and superior transfection of SK-OV-3 cell line compared to the commercially available DOTAP and branched polyethyleneimine (25 kDa bPEI). PMID:28239294

  7. Hemizygosity Enhances Purifying Selection: Lack of Fast-Z Evolution in Two Satyrine Butterflies.

    PubMed

    Rousselle, Marjolaine; Faivre, Nicolas; Ballenghien, Marion; Galtier, Nicolas; Nabholz, Benoit

    2016-10-23

    The fixation probability of a recessive beneficial mutation is increased on the X or Z chromosome, relative to autosomes, because recessive alleles carried by X or Z are exposed to selection in the heterogametic sex. This leads to an increased dN/dS ratio on sex chromosomes relative to autosomes, a pattern called the "fast-X" or "fast-Z" effect. Besides positive selection, the strength of genetic drift and the efficacy of purifying selection, which affect the rate of molecular evolution, might differ between sex chromosomes and autosomes. Disentangling the complex effects of these distinct forces requires the genome-wide analysis of polymorphism, divergence and gene expression data in a variety of taxa. Here we study the influence of hemizygosity of the Z chromosome in Maniola jurtina and Pyronia tithonus, two species of butterflies (Lepidoptera, Nymphalidae, Satyrinae). Using transcriptome data, we compare the strength of positive and negative selection between Z and autosomes accounting for sex-specific gene expression. We show that M. jurtina and P. tithonus do not experience a faster, but rather a slightly slower evolutionary rate on the Z than on autosomes. Our analysis failed to detect a significant difference in adaptive evolutionary rate between Z and autosomes, but comparison of male-biased, unbiased and female-biased Z-linked genes revealed an increased efficacy of purifying selection against recessive deleterious mutations in female-biased Z-linked genes. This probably contributes to the lack of fast-Z evolution of satyrines. We suggest that the effect of hemizygosity on the fate of recessive deleterious mutations should be taken into account when interpreting patterns of molecular evolution in sex chromosomes vs. autosomes.

  8. Transfection Studies with Colloidal Systems Containing Highly Purified Bipolar Tetraether Lipids from Sulfolobus acidocaldarius.

    PubMed

    Engelhardt, Konrad H; Pinnapireddy, Shashank Reddy; Baghdan, Elias; Jedelská, Jarmila; Bakowsky, Udo

    2017-01-01

    Lipid vectors are commonly used to facilitate the transfer of nucleic acids into mammalian cells. In this study, two fractions of tetraether lipids from the archaea Sulfolobus acidocaldarius were extracted and purified using different methods. The purified lipid fractions polar lipid fraction E (PLFE) and hydrolysed glycerol-dialkyl-nonitol tetraether (hGDNT) differ in their structures, charge, size, and miscibility from conventional lipids. Liposomes were prepared by mixing tetraether lipids with cholesterol (CH) and 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) resulting in stable vectors for gene delivery. Lipoplexes were prepared by complexation of liposomes with a luciferase expressing plasmid (pCMV-luc) at certain nitrogen-to-phosphorus (N/P) ratios and optimised for the transient transfection of ovarian adenocarcinoma cells (SK-OV-3). Complexation efficacy was investigated by gel-red fluorescence assay. Biophysical properties, like size, surface charge, and morphology, were investigated by differential light scattering (DLS), atomic force microscopy (AFM), and scanning electron microscopy (Cryo-SEM), respectively, revealing structural differences between liposomes and lipoplexes. A range of stable transfecting agents containing tetraether lipids were obtained by incorporating 5 mol% of tetraether lipids. Lipoplexes showed a decrease in free gel-red with increasing N/P ratios indicating efficient incorporation of plasmid DNA (pDNA) and remarkable stability. Transfection experiments of the lipoplexes revealed successful and superior transfection of SK-OV-3 cell line compared to the commercially available DOTAP and branched polyethyleneimine (25 kDa bPEI).

  9. Anti-HCV Activity from Semi-purified Methanolic Root Extracts of Valeriana wallichii.

    PubMed

    Ganta, Krishna Kumar; Mandal, Anirban; Debnath, Sukalyani; Hazra, Banasri; Chaubey, Binay

    2017-03-01

    Hepatitis C virus (HCV) is a serious global health problem affecting approximately 130-150 million individuals. Presently available direct-acting anti-HCV drugs have higher barriers to resistance and also improved success rate; however, cost concerns limit their utilization, especially in developing countries like India. Therefore, development of additional agents to combat HCV infection is needed. In the present study, we have evaluated anti-HCV potential of water, chloroform, and methanol extracts from roots of Valeriana wallichii, a traditional Indian medicinal plant. Huh-7.5 cells infected with J6/JFH chimeric HCV strain were treated with water, chloroform, and methanol extracts at different concentrations. Semi-quantitative reverse transcription polymerase chain reaction result demonstrated that methanolic extract showed reduction in HCV replication. The methanolic extract was fractionated by thin layer chromatography, and the purified fractions (F1, F2, F3, and F4) were checked for anti-HCV activity. Significant viral inhibition was noted only in F4 fraction. Further, intrinsic fluorescence assay of purified HCV RNA-dependent RNA polymerase NS5B in the presence of F4 resulted in sharp quenching of intrinsic fluorescence with increasing amount of plant extract. Our results indicated that methanolic extract of V. wallichii and its fraction (F4) inhibited HCV by binding with HCV NS5B protein. The findings would be further investigated to identify the active principle/lead molecule towards development of complementary and alternative therapeutics against HCV. Copyright © 2017 John Wiley & Sons, Ltd.

  10. In vitro fermentation by human faecal bacteria of total and purified dietary fibres from brown seaweeds.

    PubMed

    Michel, C; Lahaye, M; Bonnet, C; Mabeau, S; Barry, J L

    1996-02-01

    The in vitro degradation of dietary fibre from three brown seaweeds (Himanthalia elongata, Laminaria digitata and Undaria pinnatifida) was studied, using human faecal flora. Two sets of fibre were tested: (1) total algal fibres extracted from the whole algae, mainly composed of alginates, and (2) purified fibres (sulphated fucans, Na-alginates and laminarans) representative of those contained in the whole brown algae. Mannuronate, one algal component, was also investigated. Substrate disappearance and short-chain fatty acid (SCFA) production were monitored after 6, 12 and 24 h fermentation. Gas production was followed hourly during the first 9 h and then at 12 and 24 h. Sugarbeet fibre was used as a fermentation reference substrate. According to the fermentative indices used, most of each of the total algal fibres disappeared after 24 h (range 60-76%) but, unlike the reference substrate, they were not completely metabolized to SCFA (range 47-62%). Among the purified algal fibres, disappearance of laminarans was approximately 90% and metabolism to SCFA was approximately 85% in close agreement with the fermentation pattern of reference fibres. Sulphated fucans were not degraded. Na-alginates exhibited a fermentation pattern quite similar to those of the whole algal fibres with a more pronounced discrepancy between disappearance and production of SCFA: disappearance was approximately 83% but metabolism was only approximately 57%. Mannuronate was slowly fermented but its metabolism corresponded to its disappearance from the fermentative medium. Thus, the characteristic fermentation pattern of the total fibres from the three brown algae investigated was attributed to the peculiar fermentation of alginates, and mannuronate was shown not to be directly involved.

  11. Recombinant Passenger Proteins Can Be Conveniently Purified by One-Step Affinity Chromatography.

    PubMed

    Wang, Hua-zhen; Chu, Zhi-zhan; Chen, Chang-chao; Cao, Ao-cheng; Tong, Xin; Ouyang, Can-bin; Yuan, Qi-hang; Wang, Mi-nan; Wu, Zhong-kun; Wang, Hai-hong; Wang, Sheng-bin

    2015-01-01

    Fusion tag is one of the best available tools to date for enhancement of the solubility or improvement of the expression level of recombinant proteins in Escherichia coli. Typically, two consecutive affinity purification steps are often necessitated for the purification of passenger proteins. As a fusion tag, acyl carrier protein (ACP) could greatly increase the soluble expression level of Glucokinase (GlcK), α-Amylase (Amy) and GFP. When fusion protein ACP-G2-GlcK-Histag and ACP-G2-Amy-Histag, in which a protease TEV recognition site was inserted between the fusion tag and passenger protein, were coexpressed with protease TEV respectively in E. coli, the efficient intracellular processing of fusion proteins was achieved. The resulting passenger protein GlcK-Histag and Amy-Histag accumulated predominantly in a soluble form, and could be conveniently purified by one-step Ni-chelating chromatography. However, the fusion protein ACP-GFP-Histag was processed incompletely by the protease TEV coexpressed in vivo, and a large portion of the resulting target protein GFP-Histag aggregated in insoluble form, indicating that the intracellular processing may affect the solubility of cleaved passenger protein. In this context, the soluble fusion protein ACP-GFP-Histag, contained in the supernatant of E. coli cell lysate, was directly subjected to cleavage in vitro by mixing it with the clarified cell lysate of E. coli overexpressing protease TEV. Consequently, the resulting target protein GFP-Histag could accumulate predominantly in a soluble form, and be purified conveniently by one-step Ni-chelating chromatography. The approaches presented here greatly simplify the purification process of passenger proteins, and eliminate the use of large amounts of pure site-specific proteases.

  12. Toxicity of Raw and Purified Single-Walled Carbon Nanotubes in Rat's Lung Epithelial and Cervical Cancer Cells.

    PubMed

    Goornavar, Virupaxi; Biradar, Santoshkumar; Ezeagwu, Christian; Ezeagwu, Dexter; Hall, Joseph C; Ramesh, Govindarajan T

    2015-03-01

    The increased applications of carbon nanotubes in the field of drug delivery, bioimaging and biosensors demand nanotubes to be of highest purity, free from metallic impurities and amorphous carbon. All of these sectors require a profound investigation about the toxic effects on human and the environment. Many attempts have been made to purify and surface modify the carbon nanotubes, however a detailed study on the raw and purified material has yet to be conducted. Here we present the toxicity studies of raw and the purified single-walled carbon nanotubes in rat's lung epithelial cell and cervical cancer cells (HeLa). These cells were treated with increasing concentration of 0.5 µg/mL to 50 µg/mL and the various biocompatibility assays were performed. The results showed an increased cell death with purified single-walled carbon nanotubes followed by the depletion of antioxidant levels and activation of the caspase cascade at a rapid rate compared to raw single-walled carbon nanotubes. This suggests that purified single walled carbon nanotubes are more toxic to the cells and exhibit ultra-fine particulate matter like toxicity.

  13. Low cost thermoformed solar still water purifier for D&E countries

    NASA Astrophysics Data System (ADS)

    Flendrig, L. M.; Shah, B.; Subrahmaniam, N.; Ramakrishnan, V.

    IntroductionSolar distillation mimics nature’s hydrologic water cycle by purifying water through evaporation (using solar energy) and condensation (rain). It is one of the most basic purification systems available today to obtain high quality drinking water and can remove non-volatile contamination from almost any water source. This low-tech technology should therefore be ideally suited for developing and emerging countries where sun shines in abundance. In the past century numerous designs have been realised with footprints ranging from 0.5 m 2 to thousands of square meters. Despite all efforts, this intriguing technology has not been applied widely yet. Among the challenges that remain are: (1) its low yield, (2) obtaining local commitment to operate/maintain large scale systems properly, and (3) relatively high initial investment costs. The objective of this study has been to address challenges 1 and 3 by using standard plastic thermoforming technology to realize a small scale single slope solar still for personal use (2-4 l per day) with adequate efficiency and at low production costs. Materials and methodsThe solar still consists of two parts: a basin that holds the dirty water and a transparent tilted cover onto which the clean water vapour can condense. The basin has a footprint of 1.34 m 2 and is made of a 3 mm thick sheet of black high-density polyethylene (HDPE) which is thermoformed using standard equipment for making fish-ponds. This allows for the incorporation of detailed features, like reinforcements and a clean-water collection gutter, at no extra cost. The transparent cover is made of UV stabilised low-density PE-foil which is under a slope of 10° to transport condensed water droplets to the lower located collection gutter. Throughput and purification performance were evaluated in duplicate at our Bangalore R&D facilities in India, over a short term (5 day) period. Solar radiation was measured using a Pyranometer. The system was loaded with 40 l

  14. Regioselectivity of nitroglycerin denitration by flavoprotein nitroester reductases purified from two Pseudomonas species.

    PubMed Central

    Blehert, D S; Knoke, K L; Fox, B G; Chambliss, G H

    1997-01-01

    Two species of Pseudomonas capable of utilizing nitroglycerin (NG) as a sole nitrogen source were isolated from NG-contaminated soil and identified as Pseudomonas putida II-B and P. fluorescens I-C. While 9 of 13 laboratory bacterial strains that presumably had no previous exposure to NG could degrade low concentrations of NG (0.44 mM), the natural isolates tolerated concentrations of NG that were toxic to the lab strains (1.76 mM and higher). Whole-cell studies revealed that the two natural isolates produced different mixtures of the isomers of dinitroglycerol (DNG) and mononitroglycerol (MNG). A monomeric, flavin mononucleotide-containing NG reductase was purified from each natural isolate. These enzymes catalyzed the NADPH-dependent denitration of NG, yielding nitrite. Apparent kinetic constants were determined for both reductases. The P. putida enzyme had a Km for NG of 52 +/- 4 microM, a Km for NADPH of 28 +/- 2 microM, and a Vmax of 124 +/- 6 microM x min(-1), while the P. fluorescens enzyme had a Km for NG of 110 +/- 10 microM, a Km for NADPH of 5 +/- 1 microM, and a Vmax of 110 +/- 11 microM x min(-1). Anaerobic titration experiments confirmed the stoichiometry of NADPH consumption, changes in flavin oxidation state, and multiple steps of nitrite removal from NG. The products formed during time-dependent denitration reactions were consistent with a single enzyme being responsible for the in vivo product distributions. Simulation of the product formation kinetics by numerical integration showed that the P. putida enzyme produced an approximately 2-fold molar excess of 1,2-DNG relative to 1,3-DNG. This result could be fortuitous or could possibly be consistent with a random removal of the first nitro group from either the terminal (C-1 and C-3) positions or middle (C-2) position. However, during the denitration of 1,2-DNG, a 1.3-fold selectivity for the C-1 nitro group was determined. Comparable simulations of the product distributions from the P. fluorescens

  15. Effects of purified lignin on in vitro rumen metabolism and growth performance of feedlot cattle

    PubMed Central

    Wang, Yuxi; McAllister, Tim A.; Lora, Jairo H.

    2017-01-01

    Objective The objectives were to assess the effects of purified lignin from wheat straw (sodium hydroxide dehydrated lignin; SHDL) on in vitro ruminal fermentation and on the growth performance of feedlot cattle. Methods In vitro experiments were conducted by incubating a timothy-alfalfa (50:50) forage mixture (48 h) and barley grain (24 h) with 0, 0.25, 0.5, 1.0, and 2.0 mg/mL of rumen fluid (equivalent to 0, 2, 4, 8, and 16 g SHDL/kg diet). Productions of CH4 and total gas, volatile fatty acids, ammonia, dry matter (DM) disappearance (DMD) and digestion of neutral detergent fiber (NDF) or starch were measured. Sixty Hereford-Angus cross weaned steer calves were individually fed a typical barley silage-barley grain based total mixed ration and supplemented with SHDL at 0, 4, 8, and 16 g/kg DM for 70 (growing), 28 (transition), and 121 d (finishing) period. Cattle were slaughtered at the end of the experiment and carcass traits were assessed. Results With forage, SHDL linearly (p<0.001) reduced 48-h in vitro DMD from 54.9% to 39.2%, NDF disappearance from 34.1% to 18.6% and the acetate: propionate ratio from 2.56 to 2.41, but linearly (p<0.001) increased CH4 production from 9.5 to 12.4 mL/100 mg DMD. With barley grain, SHDL linearly increased (p<0.001) 24-h DMD from74.6% to 84.5%, but linearly (p<0.001) reduced CH4 production from 5.6 to 4.2 mL/100 mg DMD and NH3 accumulation from 9.15 to 4.49 μmol/mL. Supplementation of SHDL did not affect growth, but tended (p = 0.10) to linearly reduce feed intake, and quadratically increased (p = 0.059) feed efficiency during the finishing period. Addition of SHDL also tended (p = 0.098) to linearly increase the saleable meat yield of the carcass from 52.5% to 55.7%. Conclusion Purified lignin used as feed additive has potential to improve feed efficiency for finishing feedlot cattle and carcass quality. PMID:27456424

  16. Properties of binding of partially purified glucocorticoid receptor from rat liver with glucocorticoids of different biopotencies.

    PubMed

    Izawa, M; Satoh, Y; Yoshida, A; Ichii, S

    1985-06-01

    To elucidate the relationship between binding parameters and biopotencies of glucocorticoids, we partially purified the receptor from the liver cytosol of rats in a dexamethasone-bound and unactivated form by precipitation with protamine sulfate, gel filtration and DEAE-cellulose chromatography (approximately 100-fold) and examined the interaction of the preparation with 3 glucocorticoids of different biopotencies (dexamethasone; Dex, corticosterone; Cort and prednisolone; Pred). The partially purified receptor (PPR) was stable at -20 degrees C for at least 2 months in the presence of bovine serum albumin, glycerol, molybdate and dithiothreitol. Treatment of the PPR with p-hydroxymercuribenzoate liberated the ligands and the treated PPR reassociated 3H-glucocorticoids efficiently following the addition of dithiothreitol. The reassociated PPR was bound to the DNA-cellulose after a brief heating. Metabolic activity on ligands and inactivation of the binding sites in the PPR were insignificant under the conditions used. Kd's were approximately 0.9, approximately 3 and approximately 6 nM for Dex, Cort and Pred, respectively (at 0 degree C). Relative binding affinity of ligands to the PPR which was estimated by competitions was higher in the order of triamcinolone acetonide greater than Dex greater than Cort greater than Pred greater than progesterone greater than cortexolone. Association of Dex and Cort was relatively rapid and significantly accelerated by raising the incubation temperature, while the association of Pred was slower and effects of the temperature was moderate. The rate of dissociations was also varied with ligands. The rate of dissociation of Dex was the lowest among the 3 ligands and was elevated by raising the temperature. Because the effect of temperature was more pronounced in the dissociation than in the association, apparent Ka's decreased at higher temperature. Thermodynamic examinations of glucocorticoid binding in the PPR revealed that the

  17. Kinetic characters and resistance to inhibition of crude and purified brain acetylcholinesterase of three freshwater fishes by organophosphates.

    PubMed

    Shaonan, Li; Xianchuan, Xie; Guonian, Zhu; Yajun, Tan

    2004-07-14

    Acetylcholinesterase (AChE) was purified from the brain of three fresh-water fishes, topmouth gudgeon (Pseudorasbora parva), goldfish (Carassius auratus auratus) and rainbow trout (Oncorrhychus mykiss, formerly named Salmo gairdneri) by PEG2000/phosphate-salt two phases extraction, DEAE-Sephadex A-50 and Sephadex G-200 chromatography. Kinetic characters and resistance to inhibition of crude and purified enzymes by organophosphates were then studied. Although the crude enzyme from the trout displayed a different specific activity, kinetic curve, Vmax, and sensitivity to inhibition by oxidized malathion and triazopos compared with the two cyprinoids (i.e. topmouth gudgeon and goldfish), the purified enzymes of all the three species showed no significant difference in all aspects. The result suggested a negligible intrinsic difference of brain AChEs among the tested species.

  18. A novel affinity-based method for the isolation of highly purified extracellular vesicles

    PubMed Central

    Nakai, Wataru; Yoshida, Takeshi; Diez, Diego; Miyatake, Yuji; Nishibu, Takahiro; Imawaka, Naoko; Naruse, Ken; Sadamura, Yoshifusa; Hanayama, Rikinari

    2016-01-01

    Extracellular vesicles (EVs) such as exosomes and microvesicles serve as messengers of intercellular network, allowing exchange of cellular components between cells. EVs carry lipids, proteins, and RNAs derived from their producing cells, and have potential as biomarkers specific to cell types and even cellular states. However, conventional methods (such as ultracentrifugation or polymeric precipitation) for isolating EVs have disadvantages regarding purity and feasibility. Here, we have developed a novel method for EV purification by using Tim4 protein, which specifically binds the phosphatidylserine displayed on the surface of EVs. Because the binding is Ca2+-dependent, intact EVs can be easily released from Tim4 by adding Ca2+ chelators. Tim4 purification, which we have applied to cell conditioned media and biofluids, is capable of yielding EVs of a higher purity than those obtained using conventional methods. The lower contamination found in Tim4-purified EV preparations allows more EV-specific proteins to be detected by mass spectrometry, enabling better characterization and quantification of different EV populations’ proteomes. Tim4 protein can also be used as a powerful tool for quantification of EVs in both ELISA and flow cytometry formats. Thus, the affinity of Tim4 for EVs will find abundant applications in EV studies. PMID:27659060

  19. Capping and methylation of mRNA by purified recombinant VP4 protein of bluetongue virus

    PubMed Central

    Ramadevi, N.; Burroughs, Nicholas J.; Mertens, Peter P. C.; Jones, Ian M.; Roy, Polly

    1998-01-01

    The core of bluetongue virus (BTV) is a multienzyme complex composed of two major proteins (VP7 and VP3) and three minor proteins (VP1, VP4, and VP6) in addition to the viral genome. The core is transcriptionally active and produces capped mRNA from which all BTV proteins are translated, but the relative role of each core component in the overall reaction process remains unclear. Previously we showed that the 76-kDa VP4 protein possesses guanylyltransferase activity, a necessary part of the RNA capping reaction. Here, through the use of highly purified (>95%) VP4 and synthetic core-like particles containing VP4, we have investigated the extent to which this protein is also responsible for other activities associated with cap formation. We show that VP4 catalyzes the conversion of unmethylated GpppG or in vitro-produced uncapped BTV RNA transcripts to m7GpppGm in the presence of S-adenosyl-l-methionine. Analysis of the methylated products of the reaction by HPLC identified both methyltransferase type 1 and type 2 activities associated with VP4, demonstrating that the complete BTV capping reaction is associated with this one protein. PMID:9811835

  20. HARP preferentially co-purifies with RPA bound to DNA-PK and blocks RPA phosphorylation.

    PubMed

    Quan, Jinhua; Yusufzai, Timur

    2014-05-01

    The HepA-related protein (HARP/SMARCAL1) is an ATP-dependent annealing helicase that is capable of rewinding DNA structures that are stably unwound due to binding of the single-stranded DNA (ssDNA)-binding protein Replication Protein A (RPA). HARP has been implicated in maintaining genome integrity through its role in DNA replication and repair, two processes that generate RPA-coated ssDNA. In addition, mutations in HARP cause a rare disease known as Schimke immuno-osseous dysplasia. In this study, we purified HARP containing complexes with the goal of identifying the predominant factors that stably associate with HARP. We found that HARP preferentially interacts with RPA molecules that are bound to the DNA-dependent protein kinase (DNA-PK). We also found that RPA is phosphorylated by DNA-PK in vitro, while the RPA-HARP complexes are not. Our results suggest that, in addition to its annealing helicase activity, which eliminates the natural binding substrate for RPA, HARP blocks the phosphorylation of RPA by DNA-PK.

  1. A purified diet for medaka (Oryzias latipes): refining a fish model for toxicological research

    SciTech Connect

    DeKoven, D.L.; Nunez, J.M.; Lester, S.M.; Conklin, D.E.; Marty, G.D.; Parker, L.M.; Hinton, D.E. )

    1992-04-01

    The overall nutritional adequacy of a purified casein-based diet (PC-diet) for the medaka (Oryzias latipes) was evaluated and compared with three diets: commercially available flaked fish food (FL-diet), live newly hatched Artemia (A-diet), and a combination of FL-diet plus A-diet (F/A-diet). Survival, growth, reproductive success, general and liver histopathology, and selected hepatic enzyme activities were compared in medaka from first feeding through reproductive maturity. The PC-diet proved adequate in all of the above criteria. When compared with fish fed F/A-diet, an initial lag in early growth rates (i.e., 0 to 30 days) occurred with the fish fed PC-diet. The FL-diet alone was not nutritionally adequate for medaka, resulting in poor growth, reduced reproductive success, lower survival, and emaciation. A significant number of spinal deformities (5.4%) were noted in medaka fed the F/A diet. Ethoxycoumarin 0-deethylase and glutathione S-transferase activities were monitored and a trend toward increasing activity with age was noted. This suggests that PC- and F/A-diets provide adequate nutrition for development of the xenobiotic metabolizing enzymes necessary for detoxification and activation of endogenous and foreign compounds. The PC-diet supported good survival, growth, reproduction, and normal histology. This diet provides a standardized, nutritionally adequate, and consistent alternative to undefined conventional diets and is less likely to contain the range of xenobiotics possible in whole, live food.

  2. Effectiveness of purified methylene blue in an experimental model of Mycobacterium ulcerans infection.

    PubMed

    Tian, Roger B D; Asmar, Shady; Napez, Claude; Lépidi, Hubert; Drancourt, Michel

    2017-03-01

    Mycobacterium ulcerans is responsible for Buruli ulcer, characterised by extensive, disabling ulcers. Standard treatment combining rifampicin and streptomycin exposes patients to toxicity and daily painful injections. In this study, the in vitro susceptibilities of 3 M. ulcerans strains, 1 Mycobacterium marinum strain and 18 strains representative of eleven other Mycobacterium species and subspecies to methylene blue were determined. Whilst growth of M. ulcerans was inhibited by 0.0125 g/L methylene blue, growth of all other tested strains was not inhibited by 1 g/L methylene blue. The effectiveness of methylene blue in a murine model of M. ulcerans infection was then tested. Topical treatment by brushing a methylene blue solution on the skin lesion, systemic treatment by intraperitoneal injection of methylene blue, and a combined treatment (topical and systemic) were tested. The three treatment groups exhibited a significantly lower clinical score compared with the non-treated control group (P <0.05). Moreover, subcutaneous nodules were significantly smaller in the systemic treatment group (excluding males) (3 ± 0.7 mm) compared with the other groups (P <0.05). The M. ulcerans insertion sequence IS2404 and the KR-B gene were detected in all challenged mice, but not in negative controls. The density of M. ulcerans (mycobacteria/cell) was significantly lower in the combined treatment group compared with the other groups. These data provide evidence for the effectiveness of purified methylene blue against the initial stage of Buruli ulcer.

  3. Biobleaching of industrial important dyes with peroxidase partially purified from garlic.

    PubMed

    Osuji, Akudo Chigozirim; Eze, Sabinus Oscar O; Osayi, Emmanuel Emeka; Chilaka, Ferdinand Chiemeka

    2014-01-01

    An acidic peroxidase was extracted from garlic (Allium sativum) and was partially purified threefold by ammonium sulphate precipitation, dialysis, and gel filtration chromatography using sephadex G-200. The specific activity of the enzyme increased from 4.89 U/mg after ammonium sulphate precipitation to 25.26 U/mg after gel filtration chromatography. The optimum temperature and pH of the enzyme were 50°C and 5.0, respectively. The Km and V max for H2O2 and o-dianisidine were 0.026 mM and 0.8 U/min, and 25 mM and 0.75 U/min, respectively. Peroxidase from garlic was effective in decolourizing Vat Yellow 2, Vat Orange 11, and Vat Black 27 better than Vat Green 9 dye. For all the parameters monitored, the decolourization was more effective at a pH range, temperature, H2O2 concentration, and enzyme concentration of 4.5-5.0, 50°C, 0.6 mM, and 0.20 U/mL, respectively. The observed properties of the enzyme together with its low cost of extraction (from local sources) show the potential of this enzyme for practical application in industrial wastewater treatment especially with hydrogen peroxide. These Vat dyes also exhibited potentials of acting as peroxidase inhibitors at alkaline pH range.

  4. The partial characterization of purified nitrite reductase and hydroxylamine oxidase from Nitrosomonas europaea

    PubMed Central

    Ritchie, G. A. F.; Nicholas, D. J. D.

    1974-01-01

    Nitrite reductase has been separated from cell-free extracts of Nitrosomonas and partially purified from hydroxylamine oxidase by polyacrylamide-gel electrophoresis. In its oxidized state the enzyme, which did not contain haem, had an extinction maximum at 590nm, which was abolished on reduction. Sodium diethyldithiocarbamate was a potent inhibitor of nitrite reductase. Enzyme activity was stimulated 2.5-fold when remixed with hydroxylamine oxidase, but was unaffected by mammalian cytochrome c. The enzyme also exhibited a low hydroxylamine-dependent nitrite reductase activity. The results suggest that this enzyme is similar to the copper-containing `denitrifying enzyme' of Pseudomonas denitrificans. A dithionite-reduced, 465nm-absorbing haemoprotein was associated with homogeneous preparations of hydroxylamine oxidase. The band at 465nm maximum was not reduced during the oxidation of hydroxylamine although the extinction was abolished on addition of hydroxylamine, NO2− or CO. These last-named compounds when added to the oxidized enzyme precluded the appearance of the 465nm-absorption band on addition of dithionite. Several properties of 465nm-absorbing haemoprotein are described. PMID:4154745

  5. Optoacoustic tomography in preclinical research: in vivo distribution of highly purified PEG-coated gold nanorods

    NASA Astrophysics Data System (ADS)

    Su, Richard; Liopo, Anton; Brecht, Hans-Peter; Ermilov, Sergey; Larin, Kirill; Oraevsky, Alexander A.

    2011-07-01

    We report on the optoacoustic (OA) imaging of the whole mouse body using a biocompatible contrast agent - highly purified, pegylated gold nanorods (GNR) - which has strong optical absorption in the near-infrared region and low level of toxicity. In vitro toxicity studies showed no significant change in survival rates of the cultured normal epithelium IEC-6 cells when incubated for 24 hours with up to 1 nM of GNR. In vivo toxicity studies in nude mice showed no pathological changes in liver 1 month after the IV injection of GNR with intra-body concentration around 0.25-0.50 nM. In order to study the enhancement of the OA contrast and accumulation of GNR in different tissues, we performed 3D OA imaging of live nude mice with IV-injected GNR. The enhancement of the OA contrast in comparison with the images of the untreated mice was visible starting 1 hour after the GNR injection. The OA contrast of kidneys, liver, and spleen peaked at about 2-3 days after the administration of the GNR, and then was gradually reducing.

  6. Reconstitution of a Minimal Ribosome-Associated Ubiquitination Pathway with Purified Factors

    PubMed Central

    Shao, Sichen; Hegde, Ramanujan S.

    2014-01-01

    Summary Ribosomes stalled on aberrant mRNAs engage quality control mechanisms that degrade the partially translated nascent polypeptide. Ubiquitination of the nascent protein is mediated by the E3 ligase Listerin via a mechanism involving ribosome subunit dissociation. Here, we reconstitute ribosome-associated ubiquitination with purified factors to define the minimal components and essential steps in this process. We find that the primary role of the ribosome splitting factors Hbs1, Pelota, and ABCE1 is to permit Listerin access to the nascent chain. Listerin alone can discriminate 60S- from 80S-nascent chain complexes to selectively ubiquitinate the former. Splitting factors can be bypassed by artificially removing the 40S subunit, suggesting that mere steric hindrance impedes Listerin recruitment. This was illustrated by a cryo-EM reconstruction of the 60S-Listerin complex that identifies a binding interface that clashes with the 40S ribosomal subunit. These results reveal the mechanistic logic of the core steps in a ribosome-associated quality control pathway. PMID:25132172

  7. Efficient Inactivation of Multi-Antibiotics Resistant Nosocomial Enterococci by Purified Hiracin Bacteriocin

    PubMed Central

    Hassan, Maryam; Brede, Dag Anders; Diep, Dzung B.; Nes, Ingolf F.; Lotfipour, Farzaneh; Hojabri, Zoya

    2015-01-01

    Purpose: Because of the emergence of multi-antibiotic resistant bacteria, a number of infectious diseases have become a major concern to treat in health care services worldwide. This situation is worsened by the fact that very limited progress has been made in developing new and potent antibiotics in recent years. In this context antimicrobial peptides (AMPs) represent new potential therapeutic compounds with bactericidal or bacteriostatic activity against closely related bacterial strains. Methods: In this study, a collection of enterococci (n=170) from clinical sources were investigated for their potential to inhibit multiresistant nosocomial enterococci from Iranian hospitals. Results: Four isolates produced antimicrobial peptides that inhibited all the antibiotic resistant enterococci. This included three Enterococcus faecium isolates producing combinations of enterocin A, B and L50 AB. The most potent antagonism was produced by E. faecalis HO91. Purification and subsequent characterization by MALDI-TOF MS, Edman degradation and DNA-sequencing revealed that the antimicrobial compound was Hiracin. The purified Hiracin was evaluated for antibacterial activity against 12 multiresistant enterococcal isolates from clinical samples. The results demonstrated that Hiracin is highly effective towards enterococci which were resistant even to antibiotics from four distinct classes. Conclusion: The present research addresses Hiracin as a promising alternative to conventional antibiotics in treatment of multiresistant enterococcal infections. PMID:26504762

  8. Toxicity of purified terephthalic acid manufacturing wastewater on reproductive system of male mice (Mus musculus).

    PubMed

    Zhang, Xu-Xiang; Sun, Shi-Lei; Zhang, Yan; Wu, Bing; Zhang, Zong-Yao; Liu, Bo; Yang, Liu-Yan; Cheng, Shu-Pei

    2010-04-15

    Reproductive toxicity of purified terephthalic acid (PTA) manufacturing wastewater on the male mice (Mus musculus) was investigated after 35-day intragastric perfusion treatment with the wastewater. Fluorescein diacetate and propidium iodide staining, and flow cytometry were used to assess the toxicity of PTA wastewater on spermatogenic cells. PTA wastewater induced significant variations in the relative percentages of immature haploid, diploid, tetraploid and S-phase spermatogonia. Percentage of viable spermatogenic cells was reduced from 93.1+/-2.3 in control group to 90.4+/-1.9 in the wastewater-treated group. Testicular histopathology revealed expansion of interstitial space and reduction in the number and size of Leydig cells induced by the wastewater, which was further certified by the decrease (10.6%) in relative testes weight and the increase (101.3%) in sperm shape abnormality in the wastewater-treated group. In this study, PTA wastewater was found to have reproductive toxicity on male mice, and public health problems may potentially arise from the discharge of the wastewater into the environment.

  9. Alum-type adjuvant effect of non-haemolytic saponins purified from Ilex and Passiflora spp.

    PubMed

    Silveira, F; Rossi, S; Fernández, C; Gosmann, G; Schenkel, E; Ferreira, F

    2011-12-01

    Five saponins purified from the leaves of three Ilex species (saponins 1 and 2 from I. dumosa; saponin 3 from I. argentina; saponin 4 from I. paraguariensis) and from Passiflora alata (saponin 5) were evaluated for their in vitro haemolytic activity and in vivo immunostimulatory ability in a mouse model using tetanus toxoid (TT) as a model antigen. The assayed saponins showed very weak or no haemolytic activity over the tested concentration range. Mice were immunized twice with TT formulated with pure saponins 1-5, or with a mixture of saponins from Quillaja saponaria, aluminum hydroxide gel or saline, which were used as controls. The elicited humoral response was evaluated by means of the time course of specific serum antibody levels up to day 131 post-priming (total IgG and isotypes); the cellular response was tested through a delayed-type hypersensitivity (DTH) assay. The assayed saponins, in particular saponins 3 and 5, showed an adjuvant effect similar to that of alum for all tested parameters. The immunostimulating potential of these compounds deserves further investigation, especially taking into account that some Ilex spp. and Passiflora alata are native crops of widespread use and economical importance in Latin America.

  10. Proteomic characterization of IgY preparations purified with a water dilution method.

    PubMed

    Nilsson, Elin; Hanrieder, Jörg; Bergquist, Jonas; Larsson, Anders

    2008-12-24

    Antigen-specific chicken IgY antibodies have been used for oral immunotherapy as an alternative or complement to antibiotics in several studies. The water dilution (WD) method has several advantages for purifying IgY. It is rapid, efficient, suitable for large-scale production, and nothing but water is added. The water-soluble fraction contains other proteins and lipids besides IgY. The protein content was characterized by two-dimensional gel electrophoresis (2DGE) and nanoflow liquid chromatography coupled offline to matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (nanoLC-MALDI TOF/TOF MS). Protein analysis was complicated due to the large dynamic concentration range, but 26 proteins could be identified. The relative protein concentrations in different batches were very similar according to protein patterns on 1D gels and protein concentration determinations. Thus, the purification method has a high reproducibility. The concentrations of cholesterols and triglycerides were low and should not have an effect on the plasma levels of treated patients. Purification of IgY for oral use with WD is therefore a recommended method.

  11. Liquefaction of hydrothermally pretreated wheat straw at high-solids content by purified Trichoderma enzymes.

    PubMed

    Szijártó, Nóra; Siika-aho, Matti; Sontag-Strohm, Tuula; Viikari, Liisa

    2011-01-01

    Enzymatic liquefaction was studied by measuring continuously the flowability change of high-solids lignocellulose substrates using a real time viscometric method. Hydrolysis experiments of hydrothermally pretreated wheat straw were carried out with purified enzymes from Trichoderma reesei; Cel7A, Cel6A, Cel7B, Cel5A, Cel12A and Xyn11A. Results obtained at 15% (w/w) solids revealed that endoglucanases, in particular Cel5A, are the key enzymes to rapidly reduce the viscosity of lignocellulose substrate. Cellobiohydrolases had only minor and the xylanase practically no effect on the viscosity. Efficient, fast liquefaction was obtained already at a dosage of 1.5 mg of Cel5A/gdrysolids. Partial replacement or supplementation of Cel5A by the other major hydrolytic enzymes did not improve the liquefaction. The reduction of viscosity did not correlate with the saccharification obtained in the same reaction, suggesting that efficient liquefaction is rather dependent on the site than the frequency of enzymatic cleavages.

  12. Toluene and ethylbenzene oxidation by purified naphthalene dioxygenase from Pseudomonas sp. strain NCIB 9816-4.

    PubMed

    Lee, K; Gibson, D T

    1996-09-01

    Purified naphthalene dioxygenase (NDO) from Pseudomonas sp. strain NCIB 9816-4 oxidized toluene to benzyl alcohol and benzaldehyde by reactions involving benzylic monooxygenation and dioxygen-dependent alcohol oxidation, respectively. Xylene and nitrotoluene isomers were also oxidized to substituted benzyl alcohol and benzaldehyde derivatives. NDO oxidized ethylbenzene sequentially through (S)-1-phenethyl alcohol (77% enantiomeric excess) and acetophenone to 2-hydroxyacetophenone. In addition, NDO also oxidized ethylbenzene through styrene to (R)-1-phenyl-1,2-ethanediol (74% enantiomeric excess) by reactions involving desaturation and dihydroxylation, respectively. Isotope experiments with 18O2, H2 18O, and D2O suggest that 1-phenethyl alcohol is oxidized to acetophenone by a minor reaction involving desaturation followed by tautomerization. The major reaction in the conversion of 1-phenethyl alcohol and benzyl alcohol to acetophenone and benzaldehyde, respectively, probably involves monohydroxylation to form a gem-diol intermediate which stereospecifically loses the incoming hydroxyl group to leave the carbonyl product. These results are compared with similar reactions catalyzed by cytochrome P-450.

  13. Taenia solium: immune response against oral or systemic immunization with purified recombinant calreticulin in mice.

    PubMed

    Fonseca-Coronado, Salvador; Ruiz-Tovar, Karina; Pérez-Tapia, Mayra; Mendlovic, Fela; Flisser, Ana

    2011-01-01

    Recombinant functional Taenia solium calreticulin (rTsCRT) confers different degrees of protection in the experimental model of intestinal taeniosis in hamsters. The aim of this study was to evaluate the immune response induced after oral or systemic immunization with an electroeluted rTsCRT in BALB/c mice. Oral immunization elicited high fecal IgA and the production of IL-4 and IL-5 by mesenteric lymph node cells after in vitro stimulation with rTSCRT, indicating a Th2 response. Mice subcutaneously immunized produced high amounts of serum IgG, being IgG1 (Th2-related) the predominant isotype, while in vitro stimulated spleen cells synthesized IL-4, IL-5 and also IFN-γ, indicating a mixed Th1/Th2 cellular response after systemic immunization. Our data show that purified rTsCRT induces polarized Th2 responses after oral immunization of mice, a common characteristic of protective immunity against helminths and, consequently, a desirable hallmark in the search for a vaccine.

  14. Antifungal properties of wheat histones (H1-H4) and purified wheat histone H1.

    PubMed

    De Lucca, Anthony J; Heden, Lars-Olof; Ingber, Bruce; Bhatnagar, Deepak

    2011-07-13

    Wheat ( Triticum spp.) histones H1, H2, H3, and H4 were extracted, and H1 was further purified. The effect of these histones on specific fungi that may or may not be pathogenic to wheat was determined. These fungi included Aspergillus flavus , Aspergillus fumigatus , Aspergillus niger , Fusarium oxysporum , Fusarium verticillioides , Fusarium solani , Fusarium graminearum , Penicillium digitatum , Penicillium italicum , and Greeneria uvicola . Non-germinated and germinating conidia of these fungi were bioassayed separately. The non-germinated and germinating conidia of all Fusarium species were highly susceptible to the mixture (H1-H4) as well as pure H1, with viability losses of 99-100% found to be significant (p < 0.001) at ≤10 μM or less for the histone mixture and pure H1. F. graminearum was the most sensitive to histone activity. The histones were inactive against all of the non-germinated Penicillium spp. conidia. However, they significantly reduced the viability of the germinating conidia of the Penicillium spp. conidia, with 95% loss at 2.5 μM. Non-germinated and germinating conidia viability of the Aspergillus spp. and G. uvicola were unaffected when exposed to histones up to 10 μM. Results indicate that Fusarium spp. pathogenic to wheat are susceptible to wheat histones, indicating that these proteins may be a resistance mechanism in wheat against fungal infection.

  15. Optimization of freeze drying conditions for purified pectinase from mango (Mangifera indica cv. Chokanan) peel.

    PubMed

    Mehrnoush, Amid; Mustafa, Shuhaimi; Yazid, Abdul Manap Mohd

    2012-01-01

    Response surface methodology (RSM) along with central composite design (CCD) was applied to optimize the freeze drying conditions for purified pectinase from mango (Mangifera indica cv. Chokanan) peel. The effect of pectinase content (-2.66, 62.66 mg/mL), Arabic gum (-1.21, 10.21%, w/v), and maltodextrin (0.73, 7.26%, w/v) as independent variables on activity, yield, and storage stability of freeze-dried enzyme was evaluated. Storage stability of pectinase was investigated after one week at 4 °C and yield percentage of the enzyme after encapsulation was also determined. The independent variables had the most significant (p < 0.05) effect on pectinase activity and yield of the enzyme. It was observed that the interaction effect of Arabic gum and maltodextrin improved the enzymatic properties of freeze-dried pectinase. The optimal conditions for freeze-dried pectinase from mango peel were obtained using 30 mg/mL of pectinase content, 4.5 (%, w/v) of Arabic gum, and 4 (%, w/v) of maltodextrin. Under these conditions, the maximum activity (11.12 U/mL), yield (86.4%) and storage stability (84.2%) of encapsulated pectinase were achieved.

  16. Steady state kinetic studies of purified yeast plasma membrane proton-translocating ATPase.

    PubMed

    Koland, J G; Hammes, G G

    1986-05-05

    The plasma membrane H+-ATPase from bakers' yeast was purified and reconstituted with phosphatidylserine. The steady state kinetics of ATP hydrolysis catalyzed by the H+-ATPase were studied over a wide range of Mg2+ and ATP concentrations. Whereas MgATP was the substrate hydrolyzed, excess concentrations of either Mg2+ or ATP were inhibitory. The dependence of the steady state initial velocity of ATP hydrolysis on the concentration of MgATP at a fixed concentration of Mg2+ was sigmoidal rather than hyperbolic. This precluded mechanisms involving only activation and inhibition by Mg2+ and competitive inhibition by ATP. Two alternative interpretations of these results are: 1) the enzyme possesses multiple catalytic sites which interact cooperatively; or 2) the enzyme can exist in multiple conformational states which catalyze MgATP hydrolysis by parallel pathways. The rate laws for both mechanisms are identical so that the two mechanisms cannot be distinguished on the basis of the kinetic data. The data are well fit by the rate law for these mechanisms with the inclusion of competitive inhibition by Mg2+ and ATP and an independent inhibition site for Mg2+.

  17. Antidiabetic activity of 50% ethanolic extract of Ricinus communis and its purified fractions.

    PubMed

    Shokeen, Poonam; Anand, Prachi; Murali, Y Krishna; Tandon, Vibha

    2008-11-01

    We investigated the antidiabetic activity of 50% ethanolic extract of roots of Ricinus communis (RCRE) along with its bioassay-guided purification. Five-hundred milligram per kilogram body weight appeared to be the effective dose as it caused the maximum lowering of the fasting blood glucose, both in normal as well as type 1 diabetic animals. The maximum hypoglycemic effect was always observed at the 8th h up to which the study has been conducted. Administration of the effective dose of RCRE to the diabetic rats for 20 days showed favorable effects not only on fasting blood glucose, but also on total lipid profile and liver and kidney functions on 10th and 20th day. RCRE was purified using silica gel column chromatography. Out of several different fractions tested, only one fraction (R-18) showed significant antihyperglycemic activity. RCRE seemed to have a high margin of safety as no mortality and no statistically significant difference in alkaline phosphatase, serum bilirubin, creatinine, serum glutamate oxaloacetate transaminase, serum glutamate pyruvate transaminase and total protein was observed even after the administration of the extract at a dose of 10 g/kg b.wt. Thus R. communis seems to have a promising value for the development of a potent phytomedicine for diabetes.

  18. N-terminal processing of affinity-tagged recombinant proteins purified by IMAC procedures.

    PubMed

    Mooney, Jane T; Fredericks, Dale P; Christensen, Thorkild; Bruun Schiødt, Christine; Hearn, Milton T W

    2015-07-01

    The ability of a new class of metal binding tags to facilitate the purification of recombinant proteins, exemplified by the tagged glutathione S-transferase and human growth hormone, from Escherichia coli fermentation broths and lysates has been further investigated. These histidine-containing tags exhibit high affinity for borderline metal ions chelated to the immobilised ligand, 1,4,7-triazacyclononane (tacn). The use of this tag-tacn immobilised metal ion affinity chromatography (IMAC) system engenders high selectivity with regard to host cell protein removal and permits facile tag removal from the E. coli-expressed recombinant protein. In particular, these tags were specifically designed to enable their efficient removal by the dipeptidyl aminopeptidase 1 (DAP-1), thus capturing the advantages of high substrate specificity and rates of cleavage. MALDI-TOF MS analysis of the cleaved products from the DAP-1 digestion of the recombinant N-terminally tagged proteins confirmed the complete removal of the tag within 4-12 h under mild experimental conditions. Overall, this study demonstrates that the use of tags specifically designed to target tacn-based IMAC resins offers a comprehensive and flexible approach for the purification of E. coli-expressed recombinant proteins, where complete removal of the tag is an essential prerequisite for subsequent application of the purified native proteins in studies aimed at delineating the molecular and cellular basis of specific biological processes.

  19. The purifying trend in the chromosomal integron in Vibrio cholerae strains during the seventh pandemic.

    PubMed

    Zhang, Cuicai; Pang, Bo; Zhou, Zhemin; Wang, Haiyin; Zhou, Haijian; Lu, Xin; Du, Pengcheng; Zhang, Lijuan; Li, Jie; Cui, Zhigang; Chen, Chen; Stokes, H W; Kan, Biao

    2014-08-01

    Chromosomal integron (CI) arrays in Vibrio spp. are generally large and display great variation. Here we determined the sequence of CI array in a toxigenic O139 Vibriocholerae strain and compared it with the arrays from the genome of different O1 biotypes available in GenBank. Then PCR scanning was used to determine the CI array variations in 83 epidemic O139 strains and subsequently these variations were compared with that found in toxigenic O1 El Tor strains in our previous work. Few differences were observed in the cohort of toxigenic O139 strains compared to the toxigenic O1 El Tor strains. On the basis of CI arrays, the toxigenic O1 El Tor and O139 strains isolated concurrently in recent years appear to be more similar to each other than to the O1 strains isolated in previous decades, suggesting a closer evolutionary relationship between them. Comparison of CI arrays in toxigenic O1 El Tor and O139 V. cholerae strains isolated between 1961 and 2009 revealed a purifying trend in the CI arrays in the chronological order during the seventh pandemic.

  20. Structural stability of transparent conducting films assembled from length purified single-wall carbon nanotubes

    SciTech Connect

    J. M. Harris; G. R. S. Iyer; D. O. Simien; J. A. Fagan; J. Y. Huh; J. Y. Chung; S. D. Hudson; J. Obrzut; J. F. Douglas; C. M. Stafford; E. K. Hobbie

    2011-01-01

    Single-wall carbon nanotube (SWCNT) films show significant promise for transparent electronics applications that demand mechanical flexibility, but durability remains an outstanding issue. In this work, thin membranes of length purified single-wall carbon nanotubes (SWCNTs) are uniaxially and isotropically compressed by depositing them on prestrained polymer substrates. Upon release of the strain, the topography, microstructure, and conductivity of the films are characterized using a combination of optical/fluorescence microscopy, light scattering, force microscopy, electron microscopy, and impedance spectroscopy. Above a critical surface mass density, films assembled from nanotubes of well-defined length exhibit a strongly nonlinear mechanical response. The measured strain dependence reveals a dramatic softening that occurs through an alignment of the SWCNTs normal to the direction of prestrain, which at small strains is also apparent as an anisotropic increase in sheet resistance along the same direction. At higher strains, the membrane conductivities increase due to a compression-induced restoration of conductive pathways. Our measurements reveal the fundamental mode of elasto-plastic deformation in these films and suggest how it might be suppressed.

  1. Biodegradability enhancement of purified terephthalic acid wastewater by coagulation-flocculation process as pretreatment.

    PubMed

    Karthik, Manikavasagam; Dafale, Nishant; Pathe, Pradyumna; Nandy, Tapas

    2008-06-15

    In this work, the coagulation-flocculation process was used as pretreatment for purified terephthalic acid (PTA) wastewater with the objective of improving its overall biodegradability. PTA production generates wastewaters with toxicants p-xylene [1,4-dimethyl-benzene (C8H10)], a major raw material used in the production process, along with some of the intermediates, viz., p-toluic acid, benzoic acid, 4-carboxybenzaldehyde, phthalic acid and terephthalic acid. These compounds affect the bio-oxidation process of wastewater treatment; hence removal of these constituents is necessary, prior to conventional aerobic treatment. This paper addresses the application of coagulation-flocculation process using chemical coagulants, viz., aluminium sulphate (alum), polyaluminium chloride (PAC), ferrous sulphate and ferric chloride in combination with anionic polyelectrolyte. Polyaluminium chloride (PAC) in conjunction with lime and polyelectrolyte removed about 63.1% chemical oxygen demand (COD) and 45.2% biochemical oxygen demand (BOD) from PTA wastewater. Coagulation-flocculation process coupled with aerobic bio-oxidation treatment of PTA wastewater achieved, COD & BOD removals of 97.4% and 99.4%, respectively. The biodegradability enhancement evaluated in terms of the BOD5/COD ratio, increased from 0.45 to 0.67 at the optimum conditions. The results obtained from these studies indicate that the coagulation-flocculation process could be a suitable pretreatment method in reducing toxicity of PTA wastewater whilst enhancing biodegradability for aerobic biological treatment scheme.

  2. Morphological Studies on Relaxed and Contracted Forms of Purified Pyocin Particles

    PubMed Central

    Higerd, Thomas B.; Baechler, Charles A.; Berk, Richard S.

    1969-01-01

    The bacteriocin from Pseudomonas aeruginosa, pyocin, consists of a contractile sheath and inner core reminiscent of T-even coliphage tails. Contraction of the outer sheath was found to be promoted by 0.5 m magnesium chloride, 1% Formalin, low pH, sonic treatment, and freezing or thawing or both. The contraction caused by 0.5 m magnesium chloride, however, was found to be reversible and occurred upon reduction of the salt concentration from 0.5 to 0.02 m. In addition, direct assay showed that pyocin activity was nearly proportional to the percentage of only uncontracted forms. Initial studies suggested that the adsorption of purified pyocin onto cell wall fragments from the sensitive indicator strain of P. aeruginosa occurs with the relaxed particle only and not with the contracted form. However, after adsorption, contraction occurred. Various morphological structures, such as tail fibers and base-platelike appendages, were also observed. Upon contraction, six tail fibers were observed on many particles, four of which appeared to originate from the sheath and two from the inner core. Polysheaths and polycores several hundred nanometers in length were also occasionally observed. Images PMID:4977989

  3. Enzymatic Hydrolysis of Alginate to Produce Oligosaccharides by a New Purified Endo-Type Alginate Lyase

    PubMed Central

    Zhu, Benwei; Chen, Meijuan; Yin, Heng; Du, Yuguang; Ning, Limin

    2016-01-01

    Enzymatic hydrolysis of sodium alginate to produce alginate oligosaccharides has drawn increasing attention due to its advantages of containing a wild reaction condition, excellent gel properties and specific products easy for purification. However, the efficient commercial enzyme tools are rarely available. A new alginate lyase with high activity (24,038 U/mg) has been purified from a newly isolated marine strain, Cellulophaga sp. NJ-1. The enzyme was most active at 50 °C and pH 8.0 and maintained stability at a broad pH range (6.0–10.0) and temperature below 40 °C. It had broad substrate specificity toward sodium alginate, heteropolymeric MG blocks (polyMG), homopolymeric M blocks (polyM) and homopolymeric G blocks (polyG), and possessed higher affinity toward polyG (15.63 mM) as well as polyMG (23.90 mM) than polyM (53.61 mM) and sodium alginate (27.21 mM). The TLC and MS spectroscopy analysis of degradation products suggested that it completely hydrolyzed sodium alginate into oligosaccharides of low degrees of polymerization (DPs). The excellent properties would make it a promising tool for full use of sodium alginate to produce oligosaccharides. PMID:27275826

  4. Oligomerization process of the hemolytic lectin CEL-III purified from a sea cucumber, Cucumaria echinata.

    PubMed

    Kuwahara, Hiromiki; Yamasaki, Takayuki; Hatakeyama, Tomomitsu; Aoyagi, Haruhiko; Fujisawa, Tetsuro

    2002-05-01

    CEL-III is a Ca(2+)-dependent lectin purified from a sea cucumber, Cucumaria echinata. This protein exhibits strong hemolytic activity as well as cytotoxicity toward some cultured cell lines. Hemolysis is caused by CEL-III oligomers formed in the cell membrane after binding to specific carbohydrate chains on the cell surface. We have found that the oligomerization of CEL-III is also induced by the binding of simple carbohydrates, such as lactose, in aqueous solution under high pH and high ionic strength conditions. From gel filtration analysis of the oligomerization of CEL-III, it was found that the formation of the CEL-III oligomer is effectively induced by the binding of lactose and lactulose, disaccharides containing a beta-galactoside structure. Electron micrographs of the resulting oligomers revealed them to exist as particles with a size of approximately 20-30 nm. The oligomerization process required more than 1 h, which is consistent with the increase in surface hydrophobicity as measured using a fluorescent probe, 8-anilinonaphthalene-1-sulfonate. However, a change in the far-UV CD spectra as well as small-angle X-ray scattering occurred within a few minutes, suggesting that a structural change in the protein takes place rapidly, but the following growth of the oligomer is a much slower process.

  5. Cutin-derived CuO reaction products from purified cuticles and tree leaves

    NASA Astrophysics Data System (ADS)

    Goñi, Miguel A.; Hedges, John I.

    1990-11-01

    Long chain (C 16-C 18) hydroxy fatty acids are obtained among the nonlignin-derived reaction products from the CuO oxidation of a variety of geochemical samples. In order to investigate the origin of these acids, the CuO reaction products of isolated cuticles and whole leaves were investigated. The reaction products from the CuO oxidation of purified apple ( Malus pumila) cuticle include 16-hydroxy-hexadecanoic acid, 10,16-dihydroxyhexadecanoic acid, 9,10,18-trihydroxyoctadec-12-enoic acid, and 9,10,18-trihydroxyoctadecanoic acid as major components. The distribution of these cutin-derived CuO reaction products is similar to the monomer compositions deduced from traditional methods of cutin analysis. Oxidation of whole English Holly ( Ilex aquifolium) leaves yields cutin-derived acidic reaction products (in addition to lignin-derived phenols) similar to those obtained from oxidation of the corresponding isolated cuticles, indicating that CuO oxidation of bulk plant tissue is a viable procedure of cutin analysis in geochemical applications.

  6. Characterization of a purified decolorizing detergent-stable peroxidase from Streptomyces griseosporeus SN9.

    PubMed

    Rekik, Hatem; Nadia, Zaraî Jaouadi; Bejar, Wacim; Kourdali, Sidali; Belhoul, Mouna; Hmidi, Maher; Benkiar, Amina; Badis, Abdelmalek; Sallem, Naim; Bejar, Samir; Jaouadi, Bassem

    2015-02-01

    A novel extracellular lignin peroxidase (called LiP-SN) was produced and purified from a newly isolated Streptomyces griseosporeus strain SN9. The findings revealed that the pure enzyme was a monomeric protein with an estimated molecular mass of 43 kDa and a Reinheitzahl value of 1.63. The 19 N-terminal residue sequence of LiP-SN showed high homology with those of Streptomyces peroxidases. Its optimum pH and temperature were pH 8.5 and 65 °C, respectively. The enzyme was inhibited by sodium azide and potassium cyanide, suggesting the presence of heme components in its tertiary structure. Its catalytic efficiency was higher than that of the peroxidase from Streptomyces albidoflavus strain TN644. Interestingly, LiP-SN showed marked dye-decolorization efficiency and stability toward denaturing, oxidizing, and bleaching agents, and compatibility with EcoVax and Dipex as laundry detergents for 48 h at 40 °C. These properties make LiP-SN a potential candidate for future applications in distaining synthetic dyes and detergent formulations.

  7. Optimization of Freeze Drying Conditions for Purified Pectinase from Mango (Mangifera indica cv. Chokanan) Peel

    PubMed Central

    Mehrnoush, Amid; Mustafa, Shuhaimi; Yazid, Abdul Manap Mohd

    2012-01-01

    Response surface methodology (RSM) along with central composite design (CCD) was applied to optimize the freeze drying conditions for purified pectinase from mango (Mangifera indica cv. Chokanan) peel. The effect of pectinase content (−2.66, 62.66 mg/mL), Arabic gum (−1.21, 10.21%, w/v), and maltodextrin (0.73, 7.26%, w/v) as independent variables on activity, yield, and storage stability of freeze-dried enzyme was evaluated. Storage stability of pectinase was investigated after one week at 4 °C and yield percentage of the enzyme after encapsulation was also determined. The independent variables had the most significant (p < 0.05) effect on pectinase activity and yield of the enzyme. It was observed that the interaction effect of Arabic gum and maltodextrin improved the enzymatic properties of freeze-dried pectinase. The optimal conditions for freeze-dried pectinase from mango peel were obtained using 30 mg/mL of pectinase content, 4.5 (%, w/v) of Arabic gum, and 4 (%, w/v) of maltodextrin. Under these conditions, the maximum activity (11.12 U/mL), yield (86.4%) and storage stability (84.2%) of encapsulated pectinase were achieved. PMID:22489134

  8. Time-resolved fluorescence spectroscopic study of flavin fluorescence in purified enzymes of bioluminescent bacteria

    NASA Astrophysics Data System (ADS)

    Vetrova, Elena; Kudryasheva, N.; Cheng, K.

    2006-10-01

    Time-resolved fluorescence intensity and anisotropy decay measurements have been used to study the environment and rotational mobility of endogenous flavin in two purified enzymes of bioluminescent bacteria, Luciferase from Photobacterium leiognathi and NAD(P)H:FMN-oxidoreductase from Vibrio fischeri. We compared the time-resolved fluorescence parameters, intensity decay lifetimes, rotational correlation times, and their fractional contribution, of the endogeneous flavin fluorescence in each of the two enzymes in the presence or absence of quinones of different structures and redox potentials. The endogeneous flavin exhibited multi-exponential decay characteristics as compared to a single decay lifetime of around 5 ns for free flavin, suggesting a complex and heterogeneous environment of flavin bound to the enzyme. In addition, a significant increase in the rotational correlation time and a certain degree of ordering of the molecule were observed for endogenous flavin when compared to a single and fast rotational correlation time of 150 ps of free flavin. Quinone significantly altered both the lifetime and rotational characteristics of endogenous flavin suggesting specific interactions of quinones to the endogeneous flavin in the bacterial enzyme.

  9. Structure, chain conformation, and immunomodulatory activity of the polysaccharide purified from Bacillus Calmette Guerin formulation.

    PubMed

    Liu, Wei; Wang, Hong; Yu, Juping; Liu, Yameng; Lu, Weisheng; Chai, Yin; Liu, Chao; Pan, Chun; Yao, Wenbing; Gao, Xiangdong

    2016-10-05

    A polysaccharide, coded as BDP, purified from the injection powder of Bacillus Calmette Guerin (BCG) polysaccharide and nucleic acid, was composed mainly of α-D-(1→4)-linked glucan with (1→6)-linked branches and trace amounts of fucose and mannose from the results of FT-IR, HPAEC-PAD and NMR spectrum. The Mw, Mn, Mz, and [Formula: see text] were determined to be 1.320×10(5)g/mol, 1.012×10(5)g/mol, 2.139×10(5)g/mol, and 21.8±3.2%nm by using HPSEC-MALLS, respectively. The ν value from [Formula: see text] was calculated to be 0.52±0.01, which firstly clarified that BDP existed as random coils in 0.9% NaCl aqueous solution. AFM and SEM combined with Congo-red test also revealed that the polysaccharide was irregular globular like or curly structure. Furthermore, in vitro tests on RAW264.7 murine macrophages cells revealed that BDP exhibited significant immunomodulatory activity.

  10. Functional Reconstitution and Channel Activity Measurements of Purified Wildtype and Mutant CFTR Protein

    PubMed Central

    Eckford, Paul D. W.; Li, Canhui; Bear, Christine E.

    2015-01-01

    The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a unique channel-forming member of the ATP Binding Cassette (ABC) superfamily of transporters. The phosphorylation and nucleotide dependent chloride channel activity of CFTR has been frequently studied in whole cell systems and as single channels in excised membrane patches. Many Cystic Fibrosis-causing mutations have been shown to alter this activity. While a small number of purification protocols have been published, a fast reconstitution method that retains channel activity and a suitable method for studying population channel activity in a purified system have been lacking. Here rapid methods are described for purification and functional reconstitution of the full-length CFTR protein into proteoliposomes of defined lipid composition that retains activity as a regulated halide channel. This reconstitution method together with a novel flux-based assay of channel activity is a suitable system for studying the population channel properties of wild type CFTR and the disease-causing mutants F508del- and G551D-CFTR. Specifically, the method has utility in studying the direct effects of phosphorylation, nucleotides and small molecules such as potentiators and inhibitors on CFTR channel activity. The methods are also amenable to the study of other membrane channels/transporters for anionic substrates. PMID:25867140

  11. Cryoelectron Microscopy Structure of Purified gamma-Secretase at 12 angstrom Resolution

    SciTech Connect

    Osenkowski, P.; Li, H.; Li, H.; Ye, W.; Li, D.; Aeschbach, L.; Fraering, P. C.; Wolfe, M. S.; Selkoe, D. J.

    2009-01-06

    {gamma}-Secretase, an integral membrane protein complex, catalyzes the intramembrane cleavage of the {beta}-amyloid precursor protein (APP) during the neuronal production of the amyloid {beta}-peptide. As such, the protease has emerged as a key target for developing agents to treat and prevent Alzheimer's disease. Existing biochemical studies conflict on the oligomeric assembly state of the protease complex, and its detailed structure is not known. Here, we report that purified active human {gamma}-secretase in digitonin has a total molecular mass of {approx} 230 kDa when measured by scanning transmission electron microscopy. This result supports a complex that is monomeric for each of the four component proteins. We further report the three-dimensional structure of the {gamma}-secretase complex at 12 {angstrom} resolution as obtained by cryoelectron microscopy and single-particle image reconstruction. The structure reveals several domains on the extracellular side, three solvent-accessible low-density cavities, and a potential substrate-binding surface groove in the transmembrane region of the complex.

  12. Structure and magnetic response of a residual metal catalyst in highly purified single walled carbon nanotubes.

    PubMed

    Bittova, Barbara Pacakova; Kalbac, Martin; Kubickova, Simona; Mantlikova, Alice; Mangold, Stephen; Vejpravova, Jana

    2013-04-28

    This article presents methods for detailed physical analysis of partial steps leading to the removal of residual metal catalyst nanoparticles (NPs) from single walled carbon nanotubes (SWCNTs) and options for detecting negligible amounts of metal in samples possessing diamagnetic response. Based on the previous knowledge of the composition, structure and magnetic properties of NPs included in the commercial HiPco_raw and HiPco_SP SWCNTs, the properties of remaining NPs after the multi-step purification (oxidation followed by mild acid treatment) and annealing both under static and dynamic vacuum have been investigated. Thermogravimetry, X-ray diffraction, static and dynamic magnetic property measurements and the Extended X-ray Absorption Fine Structure (EXAFS) experiments have been performed. The data provide information about the nature of the residual NPs in purified SWCNTs, which is crucial for further understanding of the purification processes and their improvement. It has been demonstrated that even if all macroscopic methods indicate a high purity of the treated sample, a non-negligible amount of the metal may still be present and the metal content has to be examined using local and element sensitive probes such as EXAFS.

  13. Steviol glycosides in purified stevia leaf extract sharing the same metabolic fate.

    PubMed

    Purkayastha, Sidd; Markosyan, Avetik; Prakash, Indra; Bhusari, Sachin; Pugh, George; Lynch, Barry; Roberts, Ashley

    2016-06-01

    The safety of steviol glycosides is based on data available on several individual steviol glycosides and on the terminal absorbed metabolite, steviol. Many more steviol glycosides have been identified, but are not yet included in regulatory assessments. Demonstration that these glycosides share the same metabolic fate would indicate applicability of the same regulatory paradigm. In vitro incubation assays with pooled human fecal homogenates, using rebaudiosides A, B, C, D, E, F and M, as well as steviolbioside and dulcoside A, at two concentrations over 24-48 h, were conducted to assess the metabolic fate of various steviol glycoside classes and to demonstrate that likely all steviol glycosides are metabolized to steviol. The data show that glycosidic side chains containing glucose, rhamnose, xylose, fructose and deoxy-glucose, including combinations of α(1-2), β-1, β(1-2), β(1-3), and β(1-6) linkages, were degraded to steviol mostly within 24 h. Given a common metabolite structure and a shared metabolic fate, safety data available for individual steviol glycosides can be used to support safety of purified steviol glycosides in general. Therefore, steviol glycosides specifications adopted by the regulatory authorities should include all steviol glycosides belonging to the five groups of steviol glycosides and a group acceptable daily intake established.

  14. Inhibition of purified lysosomal phospholipase A1 by beta-adrenoceptor blockers.

    PubMed

    Pappu, A S; Yazaki, P J; Hostetler, K Y

    1985-02-15

    Inhibition of rat liver lysosomal phospholipases is one of the main events that leads to accumulation of tissue phospholipids during drug-induced phospholipidosis. Drug inhibition of lysosomal phospholipase A may occur by direct effects of drugs on the enzyme (or substrate) or by drug-induced increases in intralysosomal pH. Although beta-adrenoceptor blockers have not been reported to cause lipid storage, they do inhibit lysosomal phospholipase A. To investigate the structural requirements for drug inhibition, we studied the effects of six beta-adrenoceptor blockers on purified rat liver lysosomal phospholipase A1. The agents studied include: propranolol, timolol, metoprolol, practolol, atenolol and the combined alpha and beta adrenoceptor blocking agent, labetalol. The drugs varied by two logs in their abilities to inhibit phospholipase A1 activity. The relative inhibitory potencies were propranolol greater than labetalol much greater than timolol greater than metoprolol much greater than practolol greater than atenolol. Our studies identify drug hydrophobicity as a key determinant for phospholipase A1 inhibition. A strong negative correlation was noted between the octanol/water partition coefficients and IC50 for phospholipase inhibition (r = -0.91). The ability of propranolol to inhibit phospholipase A1 was identical for the d, l and the d and l stereoisomers.

  15. Biobleaching of Industrial Important Dyes with Peroxidase Partially Purified from Garlic

    PubMed Central

    Osuji, Akudo Chigozirim; Eze, Sabinus Oscar O.; Osayi, Emmanuel Emeka; Chilaka, Ferdinand Chiemeka

    2014-01-01

    An acidic peroxidase was extracted from garlic (Allium sativum) and was partially purified threefold by ammonium sulphate precipitation, dialysis, and gel filtration chromatography using sephadex G-200. The specific activity of the enzyme increased from 4.89 U/mg after ammonium sulphate precipitation to 25.26 U/mg after gel filtration chromatography. The optimum temperature and pH of the enzyme were 50°C and 5.0, respectively. The Km and Vmax for H2O2 and o-dianisidine were 0.026 mM and 0.8 U/min, and 25 mM and 0.75 U/min, respectively. Peroxidase from garlic was effective in decolourizing Vat Yellow 2, Vat Orange 11, and Vat Black 27 better than Vat Green 9 dye. For all the parameters monitored, the decolourization was more effective at a pH range, temperature, H2O2 concentration, and enzyme concentration of 4.5–5.0, 50°C, 0.6 mM, and 0.20 U/mL, respectively. The observed properties of the enzyme together with its low cost of extraction (from local sources) show the potential of this enzyme for practical application in industrial wastewater treatment especially with hydrogen peroxide. These Vat dyes also exhibited potentials of acting as peroxidase inhibitors at alkaline pH range. PMID:25401128

  16. Evaluation of methods to concentrate and purify ocean virus communities through comparative, replicated metagenomics

    PubMed Central

    Hurwitz, Bonnie L; Deng, Li; Poulos, Bonnie T; Sullivan, Matthew B

    2013-01-01

    Viruses have global impact through mortality, nutrient cycling and horizontal gene transfer, yet their study is limited by complex methodologies with little validation. Here, we use triplicate metagenomes to compare common aquatic viral concentration and purification methods across four combinations as follows: (i) tangential flow filtration (TFF) and DNase + CsCl, (ii) FeCl3 precipitation and DNase, (iii) FeCl3 precipitation and DNase + CsCl and (iv) FeCl3 precipitation and DNase + sucrose. Taxonomic data (30% of reads) suggested that purification methods were statistically indistinguishable at any taxonomic level while concentration methods were significantly different at family and genus levels. Specifically, TFF-concentrated viral metagenomes had significantly fewer abundant viral types (Podoviridae and Phycodnaviridae) and more variability among Myoviridae than FeCl3-precipitated viral metagenomes. More comprehensive analyses using protein clusters (66% of reads) and k-mers (100% of reads) showed 50–53% of these data were common to all four methods, and revealed trace bacterial DNA contamination in TFF-concentrated metagenomes and one of three replicates concentrated using FeCl3 and purified by DNase alone. Shared k-mer analyses also revealed that polymerases used in amplification impact the resulting metagenomes, with TaKaRa enriching for ‘rare’ reads relative to PfuTurbo. Together these results provide empirical data for making experimental design decisions in culture-independent viral ecology studies. PMID:22845467

  17. Antioxidant activity of glycoprotein purified from Undaria pinnatifida measured by an in vitro digestion model.

    PubMed

    Rafiquzzaman, S M; Kim, Eun-Young; Kim, Yu-Ri; Nam, Taek-Jeong; Kong, In-Soo

    2013-11-01

    The present study was performed to investigate the chemical composition and antioxidant activity of glycoprotein purified from Undaria pinnatifida Harvey (UPGP). On SDS-PAGE, UPGP migrated as a single band with a molecular weight of approximately 10 kDa and confirmed by staining with Schiff's reagent as glycoprotein. It consists of a carbohydrate component (42.53%) and protein component (57.47%). Amino acid profile, FT-IR spectrum and enzymatic glycosylation analysis suggested that protein is linked with carbohydrate by O-glycosylation. UPGP showed dose-dependent antioxidant activities as detected by different assays before and after in vitro digestion. The IC50 values of undigested UPGP were 0.25 ± 0.03, 0.08 ± 0.005, 0.69 ± 0.12, and 0.25 ± 0.08 mg/mL for DPPH, ABTS, FRAP, and NO, respectively. Following in vitro digestion, the antioxidant activities of UPGP were decreased during the gastric phase compared to those of undigested UPGP, with an increase occurring during the duodenal phase in all assays. However, the reducing power was unchanged after in vitro digestion. Furthermore, UPGP showed protective activity against oxidative DNA damage both undigested, after saliva and duodenal phase of digestion. These results indicate that the antioxidant and DNA protection activities of UPGP may be pH-dependent and assay specific.

  18. Prescription omega-3 fatty acid products containing highly purified eicosapentaenoic acid (EPA).

    PubMed

    Brinton, Eliot A; Mason, R Preston

    2017-01-31

    The omega-3 fatty acid eicosapentaenoic acid (EPA) has multiple actions potentially conferring cardiovascular benefit, including lowering serum triglyceride (TG) and non-high-density lipoprotein cholesterol (non-HDL-C) levels and potentially reducing key steps in atherogenesis. Dietary supplements are a common source of omega-3 fatty acids in the US, but virtually all contain docosahexaenoic acid (DHA) in addition to EPA, and lipid effects differ between DHA and EPA. Contrary to popular belief, no over-the-counter omega-3 products are available in the US, only prescription products and dietary supplements. Among the US prescription omega-3 products, only one contains EPA exclusively (Vascepa); another closely related prescription omega-3 product also contains highly purified EPA, but is approved only in Japan and is provided in different capsule sizes. These high-purity EPA products do not raise low-density lipoprotein cholesterol (LDL-C) levels, even in patients with TG levels >500 mg/dL, in contrast to the increase in LDL-C levels with prescription omega-3 products that also contain DHA. The Japanese prescription EPA product was shown to significantly reduce major coronary events in hypercholesterolemic patients when added to statin therapy in the Japan EPA Lipid Intervention Study (JELIS). The effects of Vascepa on cardiovascular outcomes are being investigated in statin-treated patients with high TG levels in the Reduction of Cardiovascular Events With EPA-Intervention Trial (REDUCE-IT).

  19. CRISPR RNA binding and DNA target recognition by purified Cascade complexes from Escherichia coli.

    PubMed

    Beloglazova, Natalia; Kuznedelov, Konstantin; Flick, Robert; Datsenko, Kirill A; Brown, Greg; Popovic, Ana; Lemak, Sofia; Semenova, Ekaterina; Severinov, Konstantin; Yakunin, Alexander F

    2015-01-01

    Clustered regularly interspaced short palindromic repeats (CRISPRs) and their associated Cas proteins comprise a prokaryotic RNA-guided adaptive immune system that interferes with mobile genetic elements, such as plasmids and phages. The type I-E CRISPR interference complex Cascade from Escherichia coli is composed of five different Cas proteins and a 61-nt-long guide RNA (crRNA). crRNAs contain a unique 32-nt spacer flanked by a repeat-derived 5' handle (8 nt) and a 3' handle (21 nt). The spacer part of crRNA directs Cascade to DNA targets. Here, we show that the E. coli Cascade can be expressed and purified from cells lacking crRNAs and loaded in vitro with synthetic crRNAs, which direct it to targets complementary to crRNA spacer. The deletion of even one nucleotide from the crRNA 5' handle disrupted its binding to Cascade and target DNA recognition. In contrast, crRNA variants with just a single nucleotide downstream of the spacer part bound Cascade and the resulting ribonucleotide complex containing a 41-nt-long crRNA specifically recognized DNA targets. Thus, the E. coli Cascade-crRNA system exhibits significant flexibility suggesting that this complex can be engineered for applications in genome editing and opening the way for incorporation of site-specific labels in crRNA.

  20. Stimulation of human neutrophil leukocyte aerobic glucose metabolism by purified chemotactic factors.

    PubMed Central

    Goetzl, E J; Austen, K F

    1974-01-01

    The interaction of human neutrophils adherent to plastic petri dishes with the purified chemotactic factors C5a and kallikrein increased their rate of aerobic glycolysis 25-120% and the activity of their hexose monophosphate shunt (HMPS) 100-600%, reaching a plateau after 2 hr at 37 degrees C. The stimulation of either pathway required a chemotactically active stimulus since neither C5 nor prekallikrein or inactivated kallikrein could enhance metabolic activity. Marked suppression of the neutrophil chemotactic response by preincubation with a chemotactic factor to achieve deactivation, 5 x 10(-7) M diisopropyl fluorophosphate, or the neutrophil immobilizing factor (NIF) did not prevent the stimulation of HMPS activity or glycolysis by chemotactic factors. The metabolic inhibitors iodoacetate and 6-aminonicotinamide at concentrations which blocked enhancement of glycolysis or HMPS activity, respectively, partially suppressed the chemotactic response of neutrophils to the chemotactic factors. The capacity of a chemotactic factor to stimulate glucose metabolism of human neutrophils is associated with a maximal chemotactic response, but this stimulation is not alone sufficient for chemotaxis. Images PMID:11344574

  1. The separation of Am from lanthanides by purified Cyanex 301 extraction

    SciTech Connect

    Jing Chen; Yongjun Zhu; Rongzhou Jiao

    1996-11-01

    Separation factors of tracer amounts of Am from micro lanthanides (La, Ce, Pr, Nd, and Sm) by purified Cyanex 301 extraction in nitrate media have been determined: SF{sub Am/La} {approximately} 3500, SF{sub Am/Ce,Pr} {approximately} 1000, SF {sub Am/Nd} {approximately} 1900, and SF{sub Am/Sm}{approximately} 4500, with an average value > 2300. The distribution ratio decreases with increasing lanthanide concentration in the aqueous phase. In the presence of a macro amount of Pr + Nd(0.1 {approximately} 0.6 M) the separation factors SF{sub Am/Eu} and SF {sub Am/Pr+Nd} are about 4.7 x 10{sup 3} and 2.1 x 10{sup 3}, respectively. The results of the countercurrent fractional process show that by using three extraction stages and two scrubbing stages, >99.99% Am can be separated from a tracer amount of Eu with <0.1% extraction of Eu. Using six extraction stages, >99.99% Am and <0.6% macro amount of Pr + Nd are extracted into the organic phase.

  2. Study of Cytotoxic and Therapeutic Effects of Stable and Purified Silver Nanoparticles on Tumor Cells

    PubMed Central

    Nallathamby, Prakash D.; Xu, Xiao-Hong Nancy

    2010-01-01

    We have synthesized and purified silver nanoparticles (Ag NPs) (11.3 ± 2.3 nm) that are stable (non-aggregated) in cell culture medium and inside single living cells. We have developed new imaging methods to characterize sizes and number of single NPs in the medium and in single living cells in real-time and determine their stability (non-aggregation) in the medium and in single living cells at single NP resolution. These new approaches allow us to study toxic and therapeutic effects of single Ag NPs on tumor cells (L929, mouse fibroblast cells) with determined sizes and concentrations (doses) of NPs over time at single NP and single cell resolution. We found that Ag NPs inhibited the growth and division of tumor cells and their nuclei, at a dose and time dependent manner, showing significant inhibitory effects and abnormal cells with giant undivided nuclei or multiple nuclei beyond 12 h incubation. The results show that Ag NPs inhibited the segregation of chromosomes, but not their replications. Intracellular Ag NPs were well distributed in the cell population, and located in the nuclei and cytoplasm with higher number in the cytoplasm. This study demonstrates the possibility of using Ag NPs to inhibit the growth and division of the tumor cells and their cytotoxicity for potential therapeutic treatments, and offers a new method to count the number of single NPs in the medium for characterization their concentration and stability at single NP resolution over time. PMID:20648292

  3. All-Optical dc Nanotesla Magnetometry Using Silicon Vacancy Fine Structure in Isotopically Purified Silicon Carbide

    NASA Astrophysics Data System (ADS)

    Simin, D.; Soltamov, V. A.; Poshakinskiy, A. V.; Anisimov, A. N.; Babunts, R. A.; Tolmachev, D. O.; Mokhov, E. N.; Trupke, M.; Tarasenko, S. A.; Sperlich, A.; Baranov, P. G.; Dyakonov, V.; Astakhov, G. V.

    2016-07-01

    We uncover the fine structure of a silicon vacancy in isotopically purified silicon carbide (4H-28SiC) and reveal not yet considered terms in the spin Hamiltonian, originated from the trigonal pyramidal symmetry of this spin-3 /2 color center. These terms give rise to additional spin transitions, which would be otherwise forbidden, and lead to a level anticrossing in an external magnetic field. We observe a sharp variation of the photoluminescence intensity in the vicinity of this level anticrossing, which can be used for a purely all-optical sensing of the magnetic field. We achieve dc magnetic field sensitivity better than 100 nT /√{Hz } within a volume of 3 ×10-7m m3 at room temperature and demonstrate that this contactless method is robust at high temperatures up to at least 500 K. As our approach does not require application of radio-frequency fields, it is scalable to much larger volumes. For an optimized light-trapping waveguide of 3 mm3 , the projection noise limit is below 100 fT /√{Hz } .

  4. Purified isolation of vacuoles from Sedum alfredii leaf-derived protoplasts*

    PubMed Central

    Gao, Xiao-yu; Liao, Xing-cheng; Wu, Ruo-lai; Liu, Ting; Wang, Hai-xing; Lu, Ling-li

    2017-01-01

    This study aims to develop a method for isolating and purifying protoplasts/vacuoles from fresh leaves of the Cd hyperaccumulator plant species, Sedum alfredii. The results revealed that preheating cellulase and macerozyme at 50 °C for 5 min significantly accelerated the cell wall degradation. For the most optimal conditions for mesophyll protoplast isolation, the mixture of fresh leaves and cell lysates was followed by a 2-h–long vibration. The protoplast lysate for vacuole isolation was diluted, and 0.675 mmol/L was identified as the most appropriate 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonic acid (CHAPS) level, in which S. alfredii large vacuoles are characterized by a high metal and malic acid content. For the best vacuole purification results, we established that 0.8 mol/L was the most optimal mannitol level in the vacuole buffer in terms of vacuole protection during centrifugation, whereas a Ficoll concentration of 0.10 g/ml was adopted in the density-gradient centrifugation PMID:28071001

  5. Iron stimulation of chemiluminescence by microsomes and purified NADPH-cytochrome P-450 reductase

    SciTech Connect

    Puntarulo, S.; Clejan, L.; Palakodety, R.; Cederbaum, A.I.

    1987-05-01

    Low level chemiluminescence (CL) was measured as an assay of the steady state level of production of oxygen radicals during microsomal electron transfer. In the presence of an NADPH-generating system, antioxidant-sensitive CL was observed with isolated rat liver microsomes. Depending on the nature of the chelating agent, ferric iron markedly affected this CL. For example, ferric-EDTA inhibited, whereas ferric-ADP stimulated CL. This response to iron chelators was identical to that found when measuring microsomal lipid peroxidation, but was opposite to the catalytic effectiveness of ferric-chelates in stimulating microsomal generation of hydroxyl radicals. Similar studies were conducted with purified NADPH-cytochrome P-450 reductase in the presence of t-butyl hydroperoxide (t-BOOH). No CL was observed in the absence of added iron. The addition of ferric-EDTA or ferric-detapac stimulated production of CL, whereas ferric chloride or ferric-ATP has little or no effect. This pattern of response to iron chelates is opposite to that found with the microsomes. CL was inhibited by catalase and OH scavengers such as ethanol and DMSO but not by superoxide dismutase. Thus, CL by the reductase system appears to involve the generation of OH via a Fenton-type of reaction, and subsequent interaction of OH with t-BOOH to produce excited species.

  6. Characterization and polyanion-binding properties of purified recombinant prion protein.

    PubMed Central

    Brimacombe, D B; Bennett, A D; Wusteman, F S; Gill, A C; Dann, J C; Bostock, C J

    1999-01-01

    Certain polysulphated polyanions have been shown to have prophylactic effects on the progression of transmissible spongiform encephalopathy disease, presumably because they bind to prion protein (PrP). Until now, the difficulty of obtaining large quantities of native PrP has precluded detailed studies of these interactions. We have over-expressed murine recombinant PrP (recPrP), lacking its glycophosphoinositol membrane anchor, in modified mammalian cells. Milligram quantities of secreted, soluble and partially glycosylated protein were purified under non-denaturing conditions and the identities of mature-length aglycosyl recPrP and two cleavage fragments were determined by electrospray MS. Binding was assessed by surface plasmon resonance techniques using both direct and competitive ligand-binding approaches. recPrP binding to immobilized polyanions was enhanced by divalent metal ions. Polyanion binding was strong and showed complex association and dissociation kinetics that were consistent with ligand-directed recPrP aggregation. The differences in the binding strengths of recPrP to pentosan polysulphate and to other sulphated polyanions were found to parallel their in vivo anti-scrapie and in vitro anti-scrapie-specific PrP formation potencies. When recPrP was immobilized by capture on metal-ion chelates it was found, contrary to expectation, that the addition of polyanions promoted the dissociation of the protein. PMID:10477271

  7. Antitumor metastasis activity of pectic polysaccharide purified from the peels of Korean Citrus Hallabong.

    PubMed

    Lee, Eun Hye; Park, Hye-Ryung; Shin, Myoung-Sook; Cho, Sun Young; Choi, Hyuk-Joon; Shin, Kwang-Soon

    2014-10-13

    A polysaccharide fraction, HBE-III, was successfully purified in a high yield (40.4%) from its crude polysaccharide (HBE-0) which was prepared from pectinase hydrolysates of the peels of the Korean Citrus Hallabong. In experimental lung metastasis studies of Colon 26-M3.1 carcinoma cells, prophylactic administration of HBE-III significantly inhibited lung metastasis in a dose-dependent manner. In an in vitro cytotoxicity analysis, HBE-III (up to 1000 μg/mL) did not affect the growth of Colon 26-M3.1 cells and normal cells. HBE-III enhanced production of IL-6 and IL-12 by murine peritoneal macrophages. In an assay for natural killer (NK) cell activity, HBE-III (1000 μg/mouse, i.v.) significantly augmented NK cytotoxicity against Yac-1 tumor cells. The depletion of NK cells by injection of mouse anti-asialo GM1 serum abolished the inhibitory effect of HBE-III on lung metastasis of Colon 26-M3.1 cells. These data suggest that HBE-III may inhibit tumor metastasis via activation of macrophages and NK cells.

  8. Immunological activity of a 38-kilodalton protein purified from Mycobacterium tuberculosis.

    PubMed Central

    Young, D; Kent, L; Rees, A; Lamb, J; Ivanyi, J

    1986-01-01

    A 38-kilodalton (kDa) protein antigen from Mycobacterium tuberculosis was purified by monoclonal antibody TB71-based affinity chromatography. This molecule carries two nonoverlapping epitopes recognized by monoclonal antibodies TB71 and TB72, which are expressed substantially more strongly by M. tuberculosis than by Mycobacterium bovis. However, cross-reactive determinants between these two species were revealed on the 38-kDa protein by a rabbit anti-BCG serum. An immunoradiometric assay based on the TB71 and TB72 antibody pair specifically determined 38-kDa-antigen concentrations in mycobacterial extracts. Antibodies in sera from tuberculosis patients estimated by binding to 38-kDa-antigen-coated microtiter plates were positively correlated with TB72 competing titers. Unlike antibodies, T-cell proliferative responses to the 38-kDa protein were expressed equally by 60% of tuberculosis patients and healthy BCG-vaccinated subjects. Similarly, delayed-type hypersensitivity skin reactions were elicited in both M. tuberculosis- and M. bovis-sensitized guinea pigs. The results suggest the immunodominance of the species-specific B-cell and cross-reactive T-cell stimulatory epitopes. Images PMID:2428751

  9. [Obtaining of the affinity purified antibodies against survivin for the structure functional study of the protein].

    PubMed

    Akhidova, E V; Volkova, T D; Koroev, D O; Yakupov, I Iu; Kalintseva, M V; Zavalishina, L E; Kaplun, A P; Zharskaia, O O; Zatsepina, O V; Vol'pina, O M

    2013-01-01

    Tumor-associated protein survivin is the bifunctional protein which can participate either in cell division regulation or in apoptosis inhibition depending on its localization and structure state. The aim of this work was to obtain monospecific antibodies useful for investigation of protein structure and functional features. Six affinity purified antibodies directed to different protein regions were obtained. The ability of antibodies obtained to detect survivin in tumor cells and breast cancer tissues was studied. It was shown that antibodies to (1-22) and (95-105) survivin fragments have the highest specific activity. In western-blot antibodies to (1-22) region predominantly binds with survivin-containing complex, which may be the survivin dimer as we suppose, while antibodies to (95-105) region detects only monomeric form of the protein. Breast cancer tissues study demonstrated that survivin monomer presents only in the tumor core tissues, while survivin-containing complex is expressed both in tumor core and tumor periphery tissues. It was shown that antibodies to (1-22) fragment detect predominantly nuclear survivin, which participates in mitosis regulation, while antibodies to (95-105) fragment gave nucleoplasm and cytoplasm staining at all stages of cell cycle. Thereby antibodies obtained are the useful tool for structure-functional study of survivin.

  10. Cutin-derived CuO reaction products from purified cuticles and tree leaves

    SciTech Connect

    Goni, M.A.; Hedges, J.I. )

    1990-11-01

    Long chain (C{sub 16}-C{sub 18}) hydroxy fatty acids are obtained among the nonlignin-derived reaction products from the CuO oxidation of a variety of geochemical samples. In order to investigate the origin of these acids, the CuO reaction products of isolated cuticles and whole leaves were investigated. The reaction products from the CuO oxidation of purified apple (Malus pumila) cuticle include 16-hydroxyhexadecanoic acid, 10,16-dihydroxyhexadecanoic acid, 9,10,18-trihydroxyoctadec-12-enoic acid, and 9,10,18-trihydroxyoctadecanoic acid as major components. The distribution of these cutin-derived CuO reaction products is similar to the monomer compositions deduced from traditional methods of cutin analysis. Oxidation of whole English Holly (Ilex aquifolium) leaves yields cutin-derived acidic reaction products (in addition to lignin-derived phenols) similar to those obtained from oxidation of the corresponding isolated cuticles, indicating that CuO oxidation of bulk plant tissue is a viable procedure of cutin analysis in geochemical applications.

  11. Characterization of a partially purified Na+,K+-ATPase from dog heart.

    PubMed

    Sulakhe, P V; Elimban, V; Dhalla, N S

    1985-01-01

    Ouabain-sensitive Na+,K+-ATPase of isolated membranes represents a biochemical correlate of the "Na+ pump" that is present in intact tissue and is responsible for dissimilar distributions of Na+ and K+ across cellular plasma membranes. The enzyme has been purified from a variety of sources and its properties have been reported. Only a limited number of studies, however, deal with the cardiac Na+,K+-ATPase. We solubilized this enzyme from dog heart with deoxycholate and effected its further purification by NaI treatment. The method yielded an enzyme preparation of high specific activity (140 mumole/mg protein per hr). The following characteristics were noted: (1) pH optima of 7.4 and greater than 9.0 for ouabain-sensitive and -insensitive ATPases; (2) inhibition by Ca2+ and ouabain, the latter effect being allosteric in nature; (3) inhibition by sulfhydryl reagents (N-ethylmaleimide, p-chloromercuribenzoate) of the ouabain-sensitive ATPase but not of the -insensitive enzyme activity. All these properties resembled those seen in isolated plasma membranes (sarcolemma), suggesting that the purification procedure did not alter the properties of mono- and divalent interacting sites as well as a digitalis recognition domain of the Na+ pump. These results thus aid in further understanding the regulation of this vectorial pump that is critical in myocardial function.

  12. Isolation of highly purified type A spermatogonia from prepubertal rat testis.

    PubMed

    Morena, A R; Boitani, C; Pesce, M; De Felici, M; Stefanini, M

    1996-01-01

    We have developed a new method that allows isolation of highly purified type A spermatogonia from prepubertal rats. The procedure is based on the maximal release of spermatogonia from the seminiferous epithelium obtained by the complete enzymatic digestion of the tubular basal lamina, followed by removal of contaminating somatic cells through adhesion to plastic dishes coated with the lectin Datura stramonium agglutinin and fractionation on a discontinuous Percoll gradient. The cell suspension obtained contains up to 85% type A spermatogonia. Besides morphological criteria, the identification of germ cells and somatic cells has been performed by means of immunocytochemical markers, such as c-kit receptor, which is present only in germ cells, and vimentin, which is present only in somatic cells. All type A spermatogonia isolated were c-kit positive, thus suggesting that c-kit receptor is present in both undifferentiated and differentiating type A spermatogonia. Preliminary culture experiments demonstrate that spermatogonia survival in vitro was significantly improved by the addition of 10% fetal calf serum or horse serum to the culture medium; however, optimal culture conditions remain to be established. In vitro studies on isolated spermatogonia may provide a significant contribution toward elucidation of the mechanisms regulating spermatogonial proliferation and differentiation.

  13. Conservation and Purifying Selection of Transcribed Genes Located in a Rice Centromere[W

    PubMed Central

    Fan, Chuanzhu; Walling, Jason G.; Zhang, Jianwei; Hirsch, Cory D.; Jiang, Jiming; Wing, Rod A.

    2011-01-01

    Recombination is strongly suppressed in centromeric regions. In chromosomal regions with suppressed recombination, deleterious mutations can easily accumulate and cause degeneration of genes and genomes. Surprisingly, the centromere of chromosome8 (Cen8) of rice (Oryza sativa) contains several transcribed genes. However, it remains unclear as to what selective forces drive the evolution and existence of transcribed genes in Cen8. Sequencing of orthologous Cen8 regions from two additional Oryza species, Oryza glaberrima and Oryza brachyantha, which diverged from O. sativa 1 and 10 million years ago, respectively, revealed a set of seven transcribed Cen8 genes conserved across all three species. Chromatin immunoprecipitation analysis with the centromere-specific histone CENH3 confirmed that the sequenced orthologous regions are part of the functional centromere. All seven Cen8 genes have undergone purifying selection, representing a striking phenomenon of active gene survival within a recombination-free zone over a long evolutionary time. The coding sequences of the Cen8 genes showed sequence divergence and mutation rates that were significantly reduced from those of genes located on the chromosome arms. This suggests that Oryza has a mechanism to maintain the fidelity and functionality of Cen8 genes, even when embedded in a sea of repetitive sequences and transposable elements. PMID:21856794

  14. The antigenicity in guinea pigs and monkeys of three mycobacterial polysaccharides purified by affinity chromatography with concanavalin A.

    PubMed

    Daniel, T M

    1975-06-01

    The antigenicity of 3 polysaccharides purified from culture filtrates of Mycobacterim tuberculosis by affinity chromatography using a concanavalin A-agarose absorbent was studied. All 3 purified polysaccharides were found to be potent elicitors of delayed skin test reactions in sensitized guinea pigs and in a tuberculos monkey. This antigenicity could not be attributed to contaminating protein. Small dermal reactions were also observed in control guinea pigs. All 3 polysaccharides reacted with precipitating antibody in guinea pig sera, the antigenic specificity observed with the guinea pig sera differing from that demonstrated with reference goat antiserum. The 3 polysaccharides were also demonstrated to contain hemagglutination antigenic sites.

  15. Use of expanded bed adsorption to purify flavonoids from Ginkgo biloba L.

    PubMed

    Li, Jing; Chase, Howard A

    2009-12-11

    the column. For an expanded bed with a settled bed height of 30 cm, the loss of flavonoids in the column flow-through was about 30%. The two-step elution protocol again proved to be effective in separating the adsorbed impurities and flavonoids. More than 96% of the bound impurities were completely removed by 40% ethanol in the first elution stage and less than 4% remained in the final product eluted by 90% ethanol in the second elution stage. Also, approximately 74% of the adsorbed flavonoids on column (corresponding to 51% of the total flavonoids in the unclarified feedstock) were recovered in the product. In addition to higher recovery yield, the average process time to obtain the same amount of product was decreased in the expanded bed adsorption (EBA) process. The results suggest that the adoption of EBA procedures can greatly simplify the process flow sheet and in addition reduce the cost and time to purify flavonoids from Ginkgo biloba. These results clearly demonstrate the potential for the use of EBA to purify pharmaceuticals from plant sources.

  16. Purifying, Separating, and Concentrating Cells From a Sample Low in Biomass

    NASA Technical Reports Server (NTRS)

    Benardini, James N.; LaDuc, Myron T.; Diamond, Rochelle

    2012-01-01

    Frequently there is an inability to process and analyze samples of low biomass due to limiting amounts of relevant biomaterial in the sample. Furthermore, molecular biological protocols geared towards increasing the density of recovered cells and biomolecules of interest, by their very nature, also concentrate unwanted inhibitory humic acids and other particulates that have an adversarial effect on downstream analysis. A novel and robust fluorescence-activated cell-sorting (FACS)-based technology has been developed for purifying (removing cells from sampling matrices), separating (based on size, density, morphology), and concentrating cells (spores, prokaryotic, eukaryotic) from a sample low in biomass. The technology capitalizes on fluorescent cell-sorting technologies to purify and concentrate bacterial cells from a low-biomass, high-volume sample. Over the past decade, cell-sorting detection systems have undergone enhancements and increased sensitivity, making bacterial cell sorting a feasible concept. Although there are many unknown limitations with regard to the applicability of this technology to environmental samples (smaller cells, few cells, mixed populations), dogmatic principles support the theoretical effectiveness of this technique upon thorough testing and proper optimization. Furthermore, the pilot study from which this report is based proved effective and demonstrated this technology capable of sorting and concentrating bacterial endospore and bacterial cells of varying size and morphology. Two commercial off-the-shelf bacterial counting kits were used to optimize a bacterial stain/dye FACS protocol. A LIVE/DEAD BacLight Viability and Counting Kit was used to distinguish between the live and dead cells. A Bacterial Counting Kit comprising SYTO BC (mixture of SYTO dyes) was employed as a broad-spectrum bacterial counting agent. Optimization using epifluorescence microscopy was performed with these two dye/stains. This refined protocol was further

  17. Estimation of endogenous phosphorus loss in growing and finishing pigs fed semi-purified diets.

    PubMed

    Pettey, L A; Cromwell, G L; Lindemann, M D

    2006-03-01

    Thirty-six barrows were used in a series of 3 P-balance experiments in which growing and finishing pigs were fed highly digestible, semi-purified diets at or below the dietary available P requirement to estimate the effect of BW on endogenous P loss. Experiments 1, 2, and 3 were conducted with pigs averaging 27, 59, and 98 kg of BW, respectively. In each experiment, pigs were placed in metabolism crates and allotted by weight and litter to 3 dietary treatments. The basal diet consisted of sucrose, dextrose, cornstarch, and casein fortified with minerals (except P) and vitamins. Diets 1, 2, and 3 in Exp. 1 were the basal diet with 0, 0.078, or 0.157% added P, respectively, from monosodium phosphate. In Exp. 2 and 3, diets 1, 2, and 3 were the basal diet with 0, 0.067, and 0.134% added P, respectively, from monosodium phosphate. Within replicate, pigs were fed equal amounts of feed twice daily. Pigs were adjusted to treatments for 7 d before a 6-d, marker-to-marker collection of feces and urine. Phosphorus intakes for pigs fed the 3 diets ranged from 1.73 to 3.91 g/d in Exp. 1, from 2.18 to 5.32 g/d in Exp. 2, and from 1.96 to 6.26 g/d in Exp. 3. Fecal P excretion and P absorption increased linearly (P < 0.05) with increasing P intake. In the 3 experiments, urinary P excretion (g/d) was low for pigs fed diet 1 (0.010, 0.011, 0.019) and diet 2 (0.013, 0.058, 0.084) and was low for pigs fed diet 3 in Exp. 1 (0.037); however, urinary P was greater in pigs fed diet 3 in Exp. 2 and 3 (0.550 and 0.486, respectively). When P absorption (Y, g/d) was regressed on P intake (X, g/d) in Exp. 1, 2, and 3, the relationships were linear (P < 0.01): Y = -0.110 + 0.971X (R2 = 0.999), Y = -0.156 + 0.939X (R2 = 0.998), and Y = -0.226 + 0.8919X (R2 = 0.982), respectively. Thus, our estimates of endogenous P loss at zero P intake were 110, 156, and 226 mg/d for 27-, 59-, and 98-kg pigs, respectively. When these Y-intercepts were regressed on BW, the relationship was Y = 63.06 + 1.632X (R

  18. Transcriptomic profiling of purified patient-derived dopamine neurons identifies convergent perturbations and therapeutics for Parkinson's disease.

    PubMed

    Sandor, Cynthia; Robertson, Paul; Lang, Charmaine; Heger, Andreas; Booth, Heather; Vowles, Jane; Witty, Lorna; Bowden, Rory; Hu, Michele; Cowley, Sally A; Wade-Martins, Richard; Webber, Caleb

    2017-01-17

    While induced pluripotent stem cell (iPSC) technologies enable the study of inaccessible patient cell types, cellular heterogeneity can confound the comparison of gene expression profiles between iPSC-derived cell lines. Here, we purified iPSC-derived human dopaminergic neurons (DaNs) using the intracellular marker, tyrosine hydroxylase. Once purified, the transcriptomic profiles of iPSC-derived DaNs appear remarkably similar to profiles obtained from mature post-mortem DaNs. Comparison of the profiles of purified iPSC-derived DaNs derived from Parkinson's disease (PD) patients carrying LRRK2 G2019S variants to controls identified significant functional convergence amongst differentially-expressed (DE) genes. The PD LRRK2-G2019S associated profile was positively matched with expression changes induced by the Parkinsonian neurotoxin rotenone and opposed by those induced by clioquinol, a compound with demonstrated therapeutic efficacy in multiple PD models. No functional convergence amongst DE genes was observed following a similar comparison using non-purified iPSC-derived DaN-containing populations, with cellular heterogeneity appearing a greater confound than genotypic background.

  19. 42 CFR 84.1143 - Dust, fume, and mist air-purifying filter tests; performance requirements; general.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 42 Public Health 1 2010-10-01 2010-10-01 false Dust, fume, and mist air-purifying filter tests; performance requirements; general. 84.1143 Section 84.1143 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES OCCUPATIONAL SAFETY AND HEALTH RESEARCH AND RELATED ACTIVITIES APPROVAL...

  20. Understanding ForteBio's Sensors for High-Throughput Kinetic and Epitope Screening for Purified Antibodies and Yeast Culture Supernatant.

    PubMed

    Yu, Yao; Mitchell, Scott; Lynaugh, Heather; Brown, Michael; Nobrega, R Paul; Zhi, Xiaoyong; Sun, Tingwan; Caffry, Isabelle; Cao, Yuan; Yang, Rong; Burnina, Irina; Xu, Yingda; Estep, Patricia

    2016-01-01

    Real-time and label-free antibody screening systems are becoming more popular because of the increasing output of purified antibodies and antibody supernatant from many antibody discovery platforms. However, the properties of the biosensor can greatly affect the kinetic and epitope binning results generated by these label-free screening systems. ForteBio human-specific ProA, anti-human IgG quantitation (AHQ), anti-human Fc capture (AHC) sensors, and custom biotinylated-anti-human Fc capture (b-AHFc) sensors were evaluated in terms of loading ability, regeneration, kinetic characterization, and epitope binning with both purified IgG and IgG supernatant. AHC sensors proved unreliable for kinetic or binning assays at times, whereas AHQ sensors showed poor loading and regeneration abilities. ProA sensors worked well with both purified IgG and IgG supernatant. However, the interaction between ProA sensors and the Fab region of the IgG with VH3 germline limited the application of ProA sensors, especially in the epitope binning experiment. In an attempt to generate a biosensor type that would be compatible with a variety of germlines and sample types, we found that the custom b-AHFc sensors appeared to be robust working with both purified IgG and IgG supernatant, with little evidence of sensor-related artifacts.