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Sample records for high-molecular-weight glutenin subunits

  1. Effects of Post-Anthesis Drought and Waterlogging on Accumulation of High-Molecular-Weight Glutenin Subunits and Glutenin Macropolymers Content in Wheat Grain

    DEFF Research Database (Denmark)

    Jiang, D; Yue, H; Wollenweber, B

    2009-01-01

    Drought and flooding during grain filling have become major constraints to wheat quality and yield. The impacts of water deficits and waterlogging during the grain filling on contents of high-molecular-weight glutenin subunits (HMW-GS) and of glutenin macropolymers (GMP) in grains of the winter w...

  2. THE IMPORTANCE OF HIGH-MOLECULAR-WEIGHT GLUTENIN SUBUNITS FOR WHEAT QUALITY EVALUATION

    Directory of Open Access Journals (Sweden)

    G. Drezner

    2006-06-01

    Full Text Available High-molecular-weight glutenin subunits (HMW-GS composition was analyzed by sodium-dodecyl-sulfat-polyacrilamid-gel electrophoresis (SDS-PAGE, while the quantitative determination of total HMW-GS was obtained by reversed phase- high performance liquid chromatography (RP-HPLC. Considering HMW-GS composition, the most frequent subunits at Glu-A1 locus were N, at Glu-B1 locus 7+9 and at Glu-D1 locus 2+12. The cultivars with the GS 5+10 at Glu-D1 locus have shown better technological characteristics in contrast to cultivars with the GS 2+12. The cultivars Žitarka, Srpanjka, Barbara, Klara and Golubica in spite of presence HMW-GS 2+12 have shown very good and good technological properties because they had optimal proportions (>10% of total HMW-GS. The results of the linear correlation analysis between quality parameters and HMW-GS composition have shown significant (P<0.05 positive influence of HMW-GS (Glu-1 score on sedimentation value (r=0.55, Gluten Index (r=0.72, dough energy (r=0.61, maximum resistance (r=0.64 and resistance to extensibility ratio (r=0.58. The influence of HMW GS proportions on technological parameters, compared to HMW-GS composition, was more pronounced on protein content (r=0.82, dough development time (r=0.70, degree of softening (r=-0.90, dough energy (r=0.74 and loaf volume (r=0.65.

  3. SDS-PAGE analysis of high molecular weight glutenin subunits in SP3 from spaceflight carried wheat

    International Nuclear Information System (INIS)

    Zhang Su'na; Lv Jinyin

    2009-01-01

    The compositions of high molecular weight glutenin subunits (HMW-GS) of the third generation (SP 3 ) of two wheat varieties spaceflight carried were analyzed by SDS-PAGE. The quality score of Glu-1 of each site was calculated according to the quality rating system. The results showed that the space flight carried could result in a higher frequency of HMW-GS gene mutation. The variance frequency of HMW-GS in SP 3 of Shaan253 and Xinong1043 were 27.08% and 27.45%, and the quality score in SP 3 of Shaan253 and Xinong1043 were 7 and 6, respectively. Shaan253 SP 3 generation mutants were considered as high-quality wheat. (authors)

  4. Gluten characteristics imparting bread quality in wheats differing for high molecular weight glutenin subunits at Glu D1 locus.

    Science.gov (United States)

    Mohan, Devinder; Gupta, Raj Kumar

    2015-07-01

    High yielding genotypes differing for high molecular weight glutenin subunits at Glu D1 locus in national wheat programme of India were examined for bread loaf volume, gluten and protein contents, gluten strength, gluten index and protein-gluten ratio. Number of superior bread quality genotypes in four agro-climatically diverse zones of Indian plains was comparable in both categories of wheat i.e., 5 + 10 and 2 + 12. There wasn't any difference in average bread loaf volume and grain protein content either. 5 + 10 wheats showed better gluten strength and their gluten quality was also superior in the zones where protein content was high. 2 + 10 wheats exerted more gluten due to better protein-gluten ratio. Good bread making in 5 + 10 was derived by better gluten strength and also gluten quality in certain regions but bread quality in 2 + 12 wheats was channelized through higher gluten content as they were more efficient in extracting gluten from per unit protein. Difference in route to bread quality was apparent as gluten content and gluten strength were the key gluten attributes in 5 + 10 whereas protein content and gluten index were prominent in 2 + 12 types. Unlike 2 + 12, there was a ceiling in gluten harvest of 5 + 10 wheats as higher protein failed to deliver more gluten after some limit.

  5. [Analysis of diversity of Russian and Ukrainian bread wheat (Triticum aestivum L.) cultivars for high-molecular-weight glutenin subunits].

    Science.gov (United States)

    Dobrotvorskaia, T V; Martynov, S P

    2011-07-01

    The allelic diversity of high-moleculat-weght glutenin subunits (H WIGS) in Russian and Ukrainian bread wheat cultivars was analyzed. The diversity of spring wheat cultivars for alleles of the Glu-1 loci is characterized by medium values of the polymorphism index (polymorphism information content, PlC), and in winter wheats it varies from high at the Glu-A1 locus to low at the Glu-D1 locus. The spring and winter cultivars differ significantly in the frequencies of alleles of the glutenin loci. The combination of the Glu-A1b, Glu-B1c, and Glu-D1a alleles prevails among the spring cultivars, and the combination of the Glu-A1a, Glu-B1c, and Glu-D1d alleles prevails among the winter cultivars. The distribution of the Glu-1 alleles significantly depends on the moisture and heat supply in the region of origin of the cultivars. Drought resistance is associated with the Glu-D1a allele in the spring wheat and with the Glu-B1b allele in the winter wheat. The sources of the Glu-1 alleles were identified in the spring and wheat cultivars. The analysis of independence of the distribution of the spring and winter cultivars by the market classes and by the alleles of the HMWGS loci showed a highly significant association of the alleles of three Glu-1 loci with the market classes in foreign cultivars and independence or a weak association in the Russian and Ukrainian cultivars. This seems to be due to the absence of a statistically substantiated system of classification of the domestic cultivars on the basis of their quality.

  6. Isolation and Molecular Characterization of High Molecular Weight Glutenin Subunit Genes 1Bx13 and 1By16 from Hexaploid Wheat

    Institute of Scientific and Technical Information of China (English)

    Bin-Shuang Pang; Xue-Yong Zhang

    2008-01-01

    The high molecular weight glutenin subunit (HMW-GS) pair 1Bx13+1Byt6 are recognized to positively correlate with bread-making quality; however, their molecular data remain unknown. In order to reveal the mechanism by which 1By16 and 1Bx13 creates high quality, their open reading frames (ORFs) were amplified from common wheat Atlas66 and Jimai 20 using primers that were designed based on published sequences of HMW glutenin genes. The ORF of 1By16 was 2220bp, deduced into 738 amino acid residues with seven cysteines including 59 hexapeptides and 22 nanopeptides motifs. The ORF of 1Bx13 was 2385bp, deduced into 795 amino acid residues with four cysteines including 68 hexapeptides, 25 nanopeptides and six tripeptides motifs. We found that 1By16 was the largest y-type HMW glutenin gene described to date in common wheat. The 1By16 had 36 amino acid residues inserted in the central repetitive domain compared with 1By15. Expression in bacteria and western-blot tests confirmed that the sequence cloned was the ORF of HMW-GS 1By16, and that 1Bx13 was one of the largest 1Bx genes that have been described so far in common wheat, exhibiting a hexapeptide (PGQGQQ) insertion in the end of central repetitive domain compared with 1Bx7. A phylogenetic tree based on the deduced full-length amino acid sequence alignment of the published HMW-GS genes showed that the 1By16 was clustered with Glu-IB-2, and that the 1Bx13 was clustered with Glu-1B-1 alleles.

  7. Genetic divergence for high-molecular weight glutenin subunits (HMW-GS) in indigenous landraces and commercial cultivars of bread wheat of Pakistan.

    Science.gov (United States)

    Yasmeen, F; Khurshid, H; Ghafoor, A

    2015-05-11

    Wheat flour quality is an important consideration in the breeding and development of new cultivars. A strong association between high-molecular weight glutenin subunits (HMW-GS) and bread making quality has resulted in the widespread utilization of HMW-GS in wheat breeding. In this study, we analyzed 242 lines of wheat, including landraces from the provinces of Punjab and Baluchistan, as well as the commercial varieties of Pakistan, to determine allelic variation in the Glu-A1, Glu-B1, and Glu-D1 loci encoding HMW-GS. Higher genetic diversity was observed for HMW-GS in landraces from Baluchistan, followed by landraces collected from Punjab and then commercial varieties. Rare and uncommon subunits were observed in Glu-B1, whereas Glu-A1 was less polymorphic. However, Glu-B1 was the highest contributor to overall diversity (78%), with a total of 31 rare alleles, followed by Glu-D1 (20%) with the high quality 5+10 allele and other variants. Commercial cultivars possessed favorable alleles, potentially from indirect selection for wheat flour quality by the breeders; however, this indirect selection has decreased the pedigree base of commercial cultivars. The allelic combinations, including 2*, 5+10, and 17+18, showing high quality scores were frequent among landraces, indicating their usefulness in future crop improvement and breeding programs.

  8. Expression of wheat high molecular weight glutenin subunit 1Bx is affected by large insertions and deletions located in the upstream flanking sequences.

    Directory of Open Access Journals (Sweden)

    Yuke Geng

    Full Text Available To better understand the transcriptional regulation of high molecular weight glutenin subunit (HMW-GS expression, we isolated four Glu-1Bx promoters from six wheat cultivars exhibiting diverse protein expression levels. The activities of the diverse Glu-1Bx promoters were tested and compared with β-glucuronidase (GUS reporter fusions. Although all the full-length Glu-1Bx promoters showed endosperm-specific activities, the strongest GUS activity was observed with the 1Bx7OE promoter in both transient expression assays and stable transgenic rice lines. A 43 bp insertion in the 1Bx7OE promoter, which is absent in the 1Bx7 promoter, led to enhanced expression. Analysis of promoter deletion constructs confirmed that a 185 bp MITE (miniature inverted-repeat transposable element in the 1Bx14 promoter had a weak positive effect on Glu-1Bx expression, and a 54 bp deletion in the 1Bx13 promoter reduced endosperm-specific activity. To investigate the effect of the 43 bp insertion in the 1Bx7OE promoter, a functional marker was developed to screen 505 Chinese varieties and 160 European varieties, and only 1Bx7-type varieties harboring the 43 bp insertion in their promoters showed similar overexpression patterns. Hence, the 1Bx7OE promoter should be important tool in crop genetic engineering as well as in molecular assisted breeding.

  9. Identification of intact high molecular weight glutenin subunits from the wheat proteome using combined liquid chromatography-electrospray ionization mass spectrometry.

    Science.gov (United States)

    Lagrain, Bert; Brunnbauer, Markus; Rombouts, Ine; Koehler, Peter

    2013-01-01

    The present paper describes a method for the identification of intact high molecular weight glutenin subunits (HMW-GS), the quality determining proteins from the wheat storage proteome. The method includes isolation of HMW-GS from wheat flour, further separation of HMW-GS by reversed-phase high-performance liquid chromatography (RP-HPLC), and their subsequent molecular identification with electrospray ionization mass spectrometry using a quadrupole-time-of-flight mass analyzer. For HMW-GS isolation, wheat proteins were reduced and extracted from flour with 50% 1-propanol containing 1% dithiothreitol. HMW-GS were then selectively precipitated from the protein mixture by adjusting the 1-propanol concentration to 60%. The composition of the precipitated proteins was first evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with Coomassie staining and RP-HPLC with ultraviolet detection. Besides HMW-GS (≥65%), the isolated proteins mainly contained ω5-gliadins. Secondly, the isolated protein fraction was analyzed by liquid chromatography-mass spectrometry. Optimal chromatographic separation of HMW-GS from the other proteins in the isolated fraction was obtained when the mobile phase contained 0.1% trifluoroacetic acid as ion-pairing agent. Individual HMW-GS were then identified by determining their molecular masses from the high-resolution mass spectra and comparing these with theoretical masses calculated from amino acid sequences. Using formic acid instead of trifluoroacetic acid in the mobile phase increased protein peak intensities in the base peak mass chromatogram. This allowed the detection of even traces of other wheat proteins than HMW-GS in the isolated fraction, but the chromatographic separation was inferior with a major overlap between the elution ranges of HMW-GS and ω-gliadins. Overall, the described method allows a rapid assessment of wheat quality through the direct determination of the HMW-GS composition and offers a basis for

  10. Identification of intact high molecular weight glutenin subunits from the wheat proteome using combined liquid chromatography-electrospray ionization mass spectrometry.

    Directory of Open Access Journals (Sweden)

    Bert Lagrain

    Full Text Available The present paper describes a method for the identification of intact high molecular weight glutenin subunits (HMW-GS, the quality determining proteins from the wheat storage proteome. The method includes isolation of HMW-GS from wheat flour, further separation of HMW-GS by reversed-phase high-performance liquid chromatography (RP-HPLC, and their subsequent molecular identification with electrospray ionization mass spectrometry using a quadrupole-time-of-flight mass analyzer. For HMW-GS isolation, wheat proteins were reduced and extracted from flour with 50% 1-propanol containing 1% dithiothreitol. HMW-GS were then selectively precipitated from the protein mixture by adjusting the 1-propanol concentration to 60%. The composition of the precipitated proteins was first evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with Coomassie staining and RP-HPLC with ultraviolet detection. Besides HMW-GS (≥65%, the isolated proteins mainly contained ω5-gliadins. Secondly, the isolated protein fraction was analyzed by liquid chromatography-mass spectrometry. Optimal chromatographic separation of HMW-GS from the other proteins in the isolated fraction was obtained when the mobile phase contained 0.1% trifluoroacetic acid as ion-pairing agent. Individual HMW-GS were then identified by determining their molecular masses from the high-resolution mass spectra and comparing these with theoretical masses calculated from amino acid sequences. Using formic acid instead of trifluoroacetic acid in the mobile phase increased protein peak intensities in the base peak mass chromatogram. This allowed the detection of even traces of other wheat proteins than HMW-GS in the isolated fraction, but the chromatographic separation was inferior with a major overlap between the elution ranges of HMW-GS and ω-gliadins. Overall, the described method allows a rapid assessment of wheat quality through the direct determination of the HMW-GS composition and

  11. Coexpression of the high molecular weight glutenin subunit 1Ax1 and puroindoline improves dough mixing properties in durum wheat (Triticum turgidum L. ssp. durum).

    Science.gov (United States)

    Li, Yin; Wang, Qiong; Li, Xiaoyan; Xiao, Xin; Sun, Fusheng; Wang, Cheng; Hu, Wei; Feng, Zhijuan; Chang, Junli; Chen, Mingjie; Wang, Yuesheng; Li, Kexiu; Yang, Guangxiao; He, Guangyuan

    2012-01-01

    Wheat end-use quality mainly derives from two interrelated characteristics: the compositions of gluten proteins and grain hardness. The composition of gluten proteins determines dough rheological properties and thus confers the unique viscoelastic property on dough. One group of gluten proteins, high molecular weight glutenin subunits (HMW-GS), plays an important role in dough functional properties. On the other hand, grain hardness, which influences the milling process of flour, is controlled by Puroindoline a (Pina) and Puroindoline b (Pinb) genes. However, little is known about the combined effects of HMW-GS and PINs on dough functional properties. In this study, we crossed a Pina-expressing transgenic line with a 1Ax1-expressing line of durum wheat and screened out lines coexpressing 1Ax1 and Pina or lines expressing either 1Ax1 or Pina. Dough mixing analysis of these lines demonstrated that expression of 1Ax1 improved both dough strength and over-mixing tolerance, while expression of PINA detrimentally affected the dough resistance to extension. In lines coexpressing 1Ax1 and Pina, faster hydration of flour during mixing was observed possibly due to the lower water absorption and damaged starch caused by PINA expression. In addition, expression of 1Ax1 appeared to compensate the detrimental effect of PINA on dough resistance to extension. Consequently, coexpression of 1Ax1 and PINA in durum wheat had combined effects on dough mixing behaviors with a better dough strength and resistance to extension than those from lines expressing either 1Ax1 or Pina. The results in our study suggest that simultaneous modulation of dough strength and grain hardness in durum wheat could significantly improve its breadmaking quality and may not even impair its pastamaking potential. Therefore, coexpression of 1Ax1 and PINA in durum wheat has useful implications for breeding durum wheat with dual functionality (for pasta and bread) and may improve the economic values of durum

  12. Coexpression of the high molecular weight glutenin subunit 1Ax1 and puroindoline improves dough mixing properties in durum wheat (Triticum turgidum L. ssp. durum.

    Directory of Open Access Journals (Sweden)

    Yin Li

    Full Text Available Wheat end-use quality mainly derives from two interrelated characteristics: the compositions of gluten proteins and grain hardness. The composition of gluten proteins determines dough rheological properties and thus confers the unique viscoelastic property on dough. One group of gluten proteins, high molecular weight glutenin subunits (HMW-GS, plays an important role in dough functional properties. On the other hand, grain hardness, which influences the milling process of flour, is controlled by Puroindoline a (Pina and Puroindoline b (Pinb genes. However, little is known about the combined effects of HMW-GS and PINs on dough functional properties. In this study, we crossed a Pina-expressing transgenic line with a 1Ax1-expressing line of durum wheat and screened out lines coexpressing 1Ax1 and Pina or lines expressing either 1Ax1 or Pina. Dough mixing analysis of these lines demonstrated that expression of 1Ax1 improved both dough strength and over-mixing tolerance, while expression of PINA detrimentally affected the dough resistance to extension. In lines coexpressing 1Ax1 and Pina, faster hydration of flour during mixing was observed possibly due to the lower water absorption and damaged starch caused by PINA expression. In addition, expression of 1Ax1 appeared to compensate the detrimental effect of PINA on dough resistance to extension. Consequently, coexpression of 1Ax1 and PINA in durum wheat had combined effects on dough mixing behaviors with a better dough strength and resistance to extension than those from lines expressing either 1Ax1 or Pina. The results in our study suggest that simultaneous modulation of dough strength and grain hardness in durum wheat could significantly improve its breadmaking quality and may not even impair its pastamaking potential. Therefore, coexpression of 1Ax1 and PINA in durum wheat has useful implications for breeding durum wheat with dual functionality (for pasta and bread and may improve the economic

  13. Starch characteristics of transgenic wheat (Triticum aestivum L.) overexpressing the Dx5 high molecular weight glutenin subunit are substantially equivalent to those in nonmodified wheat.

    Science.gov (United States)

    Beckles, Diane M; Tananuwong, Kanitha; Shoemaker, Charles F

    2012-04-01

    The effects of engineering higher levels of the High Molecular Weight Glutenin Dx5 subunit on starch characteristics in transgenic wheat (Triticum aestivum L.) grain were evaluated. This is important because of the interrelationship between starch and protein accumulation in grain, the strong biotechnological interest in modulating Dx5 levels and the increasing likelihood that transgenic wheat will be commercialized in the U.S. Unintended effects of Dx5 overexpression on starch could affect wheat marketability and therefore should be examined. Two controls with native levels of Dx5 were used: (i) the nontransformed Bobwhite cultivar, and (ii) a transgenic line (Bar-D) expressing a herbicide resistant (bar) gene, and they were compared with 2 transgenic lines (Dx5G and Dx5J) containing bar and additional copies of Dx5. There were few changes between Bar-D and Dx5G compared to Bobwhite. However, Dx5J, the line with the highest Dx5 protein (×3.5) accumulated 140% more hexose, 25% less starch and the starch had a higher frequency of longer amylopectin chains. These differences were not of sufficient magnitude to influence starch functionality, because granule morphology, crystallinity, amylose-to-amylopectin ratio, and the enthalpy of starch gelatinization and the amylose-lipid complex melting were similar to the control (P > 0.05). This overall similarity was borne out by Partial Least Squares-Discriminant Function Analysis, which could not distinguish among genotypes. Collectively our data imply that higher Dx5 can affect starch accumulation and some aspects of starch molecular structure but that the starches of the Dx5 transgenic wheat lines are substantially equivalent to the controls. Transgenic manipulation of biochemical pathways is an effective way to enhance food sensory quality, but it can also lead to unintended effects. These spurious changes are a concern to Government Regulatory Agencies and to those Industries that market the product. In this study we

  14. 小麦地方品种高分子量谷蛋白亚基多样性分析%Genetic Diversity of High-Molecular-Weight Glutenin Subunit Composition in Chinese Wheat Landraces

    Institute of Scientific and Technical Information of China (English)

    徐鑫; 李小军; 张玲丽; 李秀全; 杨欣明; 李立会

    2012-01-01

    The high-molecular-weight glutenin subunit (HMW-GS) composition of 76 representative accessions of wheat land-races, collected from nine agro-ecological zones in China, were examined using sodium-dodecyl-sulphate polyacrylamide-gel electrophoresis (SDS-PAGE). The correlation between diversity indexes at Glu-1 locus and altitude, mean annual precipitation, or mean annual temperature was also analyzed. The results indicated that 19 accessions (25.0%) were heterogeneous for HMW glu-lenin subunit composition, and contained 2-4 HMW-GS compositions generally. At Glu-1 locus, a total of 14 different giutenin alleles were observed and the number of alleles at Glu-AI, Glu-BI, and Glu-DI was 2, 7, and 5, respectively. Three novel alleles were identified, consisting of two alleles at Glu-BI and one allele at Glu-DI locus. The 14 alleles resulted in 16 different HMW subunit combinations, and the combination (null, 7+8, 2+12) was the major type with the frequency of 69.7%. The genetic diversity indexes for HMW glutenin subunits varied among agro-ecological zones, and were negatively correlated with mean annual precipitation and mean annual temperature. Environmental stress is speculated as an important factor for the differentiation of diversity in wheat landraces across regions.%采用SDS-PAGE方法,对我国9个麦区的76份代表性地方品种的高分子量谷蛋白亚基(HMW-GS)组成比较分析,并探讨其与环境因素(平均海拔、年平均降雨量和年平均温度)的相关性.结果表明,25.0%的品种具有异质性,分别包含2~4种不同HMW-GS组合;在Glu-1位点共检测到14个等位变异,其中Glu-A1、Glu-B1和Glu-D1等位变异数分别为2、7和5;发现了3个新等位变异,包括Glu-B1位点2个和Glu-D1位点1个.所有等位变异构成16种不同的亚基组合类型,以(N,7+8,2+12)为主,频率为69.7%.在Glu-1位点上,不同麦区遗传多样性分布存在一定的不均衡性,年平均降雨量和年平均温度与麦区多

  15. Effects of transgene-encoded high-molecular weight glutenin proteins in wheat flour blends and sponge and dough baking

    Science.gov (United States)

    HMW glutenin subunits are the most important determinants of wheat (Triticum aestivum L.) bread-making quality, and subunit composition explains a large percentage of the variability observed between genotypes. Experiments were designed to elevate expression of a key native HMW glutenin subunit (1D...

  16. allelic variation of hmw glutenin subunits of ethiopian bread wheat

    African Journals Online (AJOL)

    journal

    High molecular weight glutenins are often effective in identifying wheat (Triticum ... There were highly significant differences between genotypes and banding ... was without deliberate selection pressure towards high Glu-1 scoring alleles ...

  17. The biosynthesis and processing of high molecular weight precursors of soybean glycinin subunits.

    Science.gov (United States)

    Barton, K A; Thompson, J F; Madison, J T; Rosenthal, R; Jarvis, N P; Beachy, R N

    1982-06-10

    The predominant storage protein of soybean seed, glycinin, is composed of two heterogeneous classes of related subunits, the acidics (Mr approximately 38,000) and the basics (Mr approximately 22,000). Immunoreaction of polypeptides translated in vitro from isolated seed mRNA using antibodies prepared against either purified acidic or basic subunit groups precipitated precursor polypeptides of Mr = 60,000 to Mr = 63,000. High pressure liquid chromatography fingerprinting of trypsin-generated fragments from in vitro synthesized precursors showed fragments specific to both acidic and basic subunits. No mature acidic or basic subunits were detected in vitro translation reactions by either immunoprecipitation or high pressure liquid chromatography fingerprinting. Pulse-labeling of cotyledons growing in culture with [3H]glycine showed rapid accumulation of label in glycinin precursors of Mr = 59,000 to Mr = 62,000. Although in vivo synthesized precursors had slightly greater electrophoretic mobility than in vitro synthesized precursors, little label initially appeared in mature glycinin subunits. After several hours of continued cotyledon growth in absence of label, precursors were processed and label accumulated in both acidic and basic subunit groups. Recombinant plasmids were prepared by reverse transcription of soybean seed mRNA, and clones which encode glycinin precursors were identified by heteroduplex-hybridization of translatable messages. Northern blot analysis of seed mRNA shows the mRNA-encoding glycinin precursors to migrate at Mr = 0.71 X 10(6) on agarose gels, corresponding to approximately 2050 nucleotides. This is sufficiently large to encode a polypeptide consisting of both a glycinin acidic and basic subunit.

  18. HMW-GS affect the properties of glutenin particles in GMP and thus flour quality

    NARCIS (Netherlands)

    Don, C.; Mann, G.; Bekes, F.; Hamer, R.J.

    2006-01-01

    Using a unique set of deletion lines, (Olympic×Gabo, varying in high molecular weight glutenin subunit (HMW-GS) composition, but with the same genetic background) it was shown that the presence of glutenin particles in glutenin macropolymer (GMP) is directly related to the presence of certain

  19. Transfer of HMW glutenin subunits from Aegilops kotschyi to wheat through radiation hybridization

    International Nuclear Information System (INIS)

    Singh, Jasmeet; Sheikh, Imran; Sharma, Prachi

    2016-01-01

    High molecular weight glutenin subunits (HMWGS) are responsible for dough elasticity and bread making quality of bread wheat. Related wild non-progenitor species, Aegilops kotschyi possesses higher molecular weight x and y glutenin subunits than the bread wheat cultivars. A wheat-Aegilops substitution line with 1U chromosome was used for the transfer of (HMWGS) of 1U to wheat by using pollen radiation hybridization approach. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiling showed different patterns of allelic variations with either the presence or absence of HMWGS, Glu-1A (1, null), Glu-1B (7, 7 + 8, 17 + 18) and Glu-1D (5 + 10, 2 + 12, null). The pollen irradiated wheat-Aegilops derivatives, B-56-1-4-2, B-56-1-4-3, B-14-1 and B-14-2 with Glu1Ux and 1Uy and absence or presence of some Glu-1A and Glu-1B HMWGS showed high micro SDS sedimentation test (MST) values while B-16-1 and B-16-2 had moderate MST values and high protein content. However, B-58-3 with transfer of Glu-1Ux + 1Uy for Glu-1D showed very low MST values indicating that Glu-1Ux + 1Uy enhance MST value only in the presence of Glu1D HMWGS. The transfer/substitution of alien HMW-GS for Glu-1A and or Glu-1B loci only can lead to improved bread making quality of wheat. (author)

  20. High molecular weight glutenin subunits of wheat : qualitative and quantitative variation in relation to bread-making quality

    NARCIS (Netherlands)

    Kolster, P.

    1992-01-01

    In view of the poor bread-making quality of the wheat grown in The Netherlands, only a small part of production is used for baking of bread. Therefore quality improvement is a major aim of plant breeding. Unfortunately, breeding for breadmaking quality is hampered by its complexity. The suitability

  1. Effect of HMM Glutenin Subunits on Wheat Quality Attributes

    Directory of Open Access Journals (Sweden)

    Daniela Horvat

    2009-01-01

    Full Text Available Glutenin is a group of polymeric gluten proteins. Glutenin molecules consist of glutenin subunits linked together with disulphide bonds and having higher (HMM-GS and lower (LMM-GS molecular mass. The main objective of this study is the evaluation of the influence of HMM-GS on flour processing properties. Seven bread wheat genotypes with contrasting quality attributes and different HMM-GS composition were analyzed during three years. The composition and quantity of HMM-GS were determined by SDS-PAGE and RP-HPLC, respectively. The quality diversity among genotypes was estimated by the analysis of wheat grain, and flour and bread quality parameters. The presence of HMM glutenin subunits 1 and 2* at Glu-A1 and the subunits 5+10 at Glu-D1 loci, as well as a higher proportion of total HMM-GS, had a positive effect on wheat quality. Cluster analysis of the three groups of data (genotype and HMM-GS, flour and bread quality, and dough rheology yielded the same hierarchical structure for the first top three levels, and similarity of the corresponding dendrograms was proved by the principal eigenvalues of the corresponding Euclidian distance matrices. The obtained similarity in classification based on essentially different types of measurements reflects strong natural association between genetic data, product quality and physical properties. Principal component analysis (PCA was applied to effectively reduce large data set into lower dimensions of latent variables amenable for the analysis. PCA analysis of the total set of data (15 variables revealed a very strong interrelationship between the variables. The first three PCA components accounted for 96 % of the total variance, which was significant to the level of 0.05 and was considered as the level of experimental error. These data imply that the quality of wheat cultivars can be contributed to HMM-GS data and should be taken into account in breeding programs assisted by computer models with the aim to

  2. Effect of shading from jointing to maturity on high molecular weight glutenin subunit accumulation and glutenin macropolymer concentration in grain of winter wheat

    DEFF Research Database (Denmark)

    Li, X.; Cai, J.; Li, H.

    2012-01-01

    the accumulation and concentration of HMW-GS in the grains. Consequently, S1 reduced falling number and SDS-sedimentation volume, while shortened dough development time (DDT) and dough stability time (DST). In contrast, S2 and S3 increased falling number, wet-gluten concentration and SDS-sedimentation volume......, and lengthened the DDT and DST. In addition, the fluctuations in accumulations of HMW-GS and GMP and most quality traits because of shading in Yangmai 158 were less than Yangmai 11. The interrelations between HMW-GS accumulation, GMP concentration and quality of grain and dough were further discussed....

  3. Low-molecular-weight glutenin subunits from the 1U genome of Aegilops umbellulata confer superior dough rheological properties and improve breadmaking quality of bread wheat.

    Science.gov (United States)

    Wang, Jian; Wang, Chang; Zhen, Shoumin; Li, Xiaohui; Yan, Yueming

    2018-04-01

    Wheat-related genomes may carry new glutenin genes with the potential for quality improvement of breadmaking. In this study, we estimated the gluten quality properties of the wheat line CNU609 derived from crossing between Chinese Spring (CS, Triticum aestivum L., 2n = 6x = 42, AABBDD) and the wheat Aegilops umbellulata (2n = 2x = 14, UU) 1U(1B) substitution line, and investigated the function of 1U-encoded low-molecular-weight glutenin subunits (LMW-GS). The main quality parameters of CNU609 were significantly improved due to introgression of the 1U genome, including dough development time, stability time, farinograph quality number, gluten index, loaf size and inner structure. Glutenin analysis showed that CNU609 and CS had the same high-molecular-weight glutenin subunit (HMW-GS) composition, but CNU609 carried eight specific 1U genome-encoded LMW-GS. The introgression of the 1U-encoded LMW-GS led to more and larger protein body formation in the CNU609 endosperm. Two new LMW-m type genes from the 1U genome, designated Glu-U3a and Glu-U3b, were cloned and characterized. Secondary structure prediction implied that both Glu-U3a and Glu-U3b encode subunits with high α-helix and β-strand content that could benefit the formation of superior gluten structure. Our results indicate that the 1U genome has superior LMW-GS that can be used as new gene resources for wheat gluten quality improvement. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  4. Effect of glutenin subunits on the baking quality of Brazilian wheat genotypes

    OpenAIRE

    Costa, Mariana Souza; Scholz, Maria Brígida dos Santos; Miranda, Martha Zavariz; Franco, Célia Maria Landi

    2017-01-01

    ABSTRACT This study aimed to evaluate the effect of the high and low molecular weight glutenin subunits on the grain traits of sixteen Brazilian wheat genotypes. Grain hardness index, milling traits, physicochemical and rheological properties of the flour, and specific volume and firmness of the bread were evaluated. Physicochemical properties of the flour were not influenced by glutenin subunits. Genotypes with subunits at the Glu-B1 (17+18 or 7+8), Glu-D1 (5+10), and Glu-A3 (b) were associa...

  5. Chemometric Analysis of High Molecular Mass Glutenin Subunits and Image Data of Bread Crumb Structure from Croatian Wheat Cultivars

    OpenAIRE

    Zorica Jurković; Rezica Sudar; Damir Magdić; Daniela Horvat; Želimir Kurtanjek

    2002-01-01

    The aim of this work is to investigate functional relationships among wheat properties, high molecular mass (weight) (HMW) glutenin subunits and bread quality produced from eleven Croatian wheat cultivars by chemometric analysis. HMW glutenin subunits were fractionated by sodium dodecylsulfate polyacrylamid gel electrophoresis (SDS-PAGE) and subsequently analysed by scanning densitometry in order to quantify HMW glutenin fractions. Wheat properties are characterised by four variables: protein...

  6. Development of haplotype-specific molecular markers for the low-molecular-weight glutenin subunits

    Science.gov (United States)

    Low-molecular-weight glutenin subunits (LMW-GSs) are one of the major components of gluten and their allelic variation has been widely associated with numerous wheat end-use quality parameters. These proteins are encoded by multigene families located at the orthologous Glu-3 loci (Glu-A3, Glu-B3 and...

  7. Acyl-CoA hydrolysis by the high molecular weight protein 1 subunit of yersiniabactin synthetase: Mutational evidence for a cascade of four acyl-enzyme intermediates during hydrolytic editing

    OpenAIRE

    Suo, Zucai; Chen, Huawei; Walsh, Christopher T.

    2000-01-01

    Yersiniabactin (Ybt) synthetase is a three-subunit, 17-domain [7 domains in high molecular weight protein (HMWP)2, 9 in HMWP1, and 1 in YbtE] enzyme producing the virulence-conferring siderophore yersiniabactin in Yersinia pestis. The 350-kDa HMWP1 subunit contains a polyketide synthase module (KS-AT-MT2-KR-ACP) and a nonribosomal peptide synthetase module (Cy3-MT3-PCP3-TE). The full-length HMWP1 was heterologously overexpressed in Escherichia coli and purified...

  8. Effect of glutenin subunits on the baking quality of Brazilian wheat genotypes

    Directory of Open Access Journals (Sweden)

    Mariana Souza Costa

    Full Text Available ABSTRACT This study aimed to evaluate the effect of the high and low molecular weight glutenin subunits on the grain traits of sixteen Brazilian wheat genotypes. Grain hardness index, milling traits, physicochemical and rheological properties of the flour, and specific volume and firmness of the bread were evaluated. Physicochemical properties of the flour were not influenced by glutenin subunits. Genotypes with subunits at the Glu-B1 (17+18 or 7+8, Glu-D1 (5+10, and Glu-A3 (b were associated with strong flours and bread with high specific volume and low firmness. The subunits at the Glu-A1 and Glu-B3 had no effect on the rheological properties of the dough and bread quality, while the subunit 2+12 at Glu-D1 negatively affected the resistance to extension, and specific volume and firmness of the bread. Specific volume and firmness of the bread were influenced by the rheological properties of the dough, while the flour protein content was not important to define wheat quality. The identification of glutenin subunits at different loci along with the rheological tests of the flour are fundamental in estimating the potential use of different materials developed in wheat breeding.

  9. Cloning and characterization of a y-type inactive HMW glutenin ...

    African Journals Online (AJOL)

    The high molecular weight glutenin subunits (HMW-GS) are key factors of the breadmaking quality of common wheat flour. In the present study, one unexpressed 1By gene from Triticum durum cultivar youmangbingmai was cloned and characterized. The results indicated that the silenced 1By gene in youmangbingmai ...

  10. Effect of high and low molecular weight glutenin subunits, and subunits of gliadin on physicochemical parameters of different wheat genotypes

    Directory of Open Access Journals (Sweden)

    Mariana Souza Costa

    2013-02-01

    Full Text Available Identification of functional properties of wheat flour by specific tests allows genotypes with appropriate characteristics to be selected for specific industrial uses. The objective of wheat breeding programs is to improve the quality of germplasm bank in order to be able to develop wheat with suitable gluten strength and extensibility for bread making. The aim of this study was to evaluate 16 wheat genotypes by correlating both glutenin subunits of high and low molecular weight and gliadin subunits with the physicochemical characteristics of the grain. Protein content, sedimentation volume, sedimentation index, and falling number values were analyzed after the grains were milled. Hectoliter weight and mass of 1000 seeds were also determined. The glutenin and gliadin subunits were separated using polyacrylamide gel in the presence of sodium dodecyl sulfate. The data were evaluated using variance analysis, Pearson's correlation, principal component analysis, and cluster analysis. The IPR 85, IPR Catuara TM, T 091015, and T 091069 genotypes stood out from the others, which indicate their possibly superior grain quality with higher sedimentation volume, higher sedimentation index, and higher mass of 1000 seeds; these genotypes possessed the subunits 1 (Glu-A1, 5 + 10 (Glu-D1, c (Glu-A3, and b (Glu-B3, with exception of T 091069 genotype that possessed the g allele instead of b in the Glu-B3.

  11. Dissemination of the highly expressed Bx7 glutenin subunit (Glu-B1al allele) in wheat as revealed by novel PCR markers and RP-HPLC.

    Science.gov (United States)

    Butow, B J; Gale, K R; Ikea, J; Juhász, A; Bedö, Z; Tamás, L; Gianibelli, M C

    2004-11-01

    Increased expression of the high molecular weight glutenin subunit (HMW-GS) Bx7 is associated with improved dough strength of wheat (Triticum aestivum L.) flour. Several cultivars and landraces of widely different genetic backgrounds from around the world have now been found to contain this so-called 'over-expressing' allelic form of the Bx7 subunit encoded by Glu-B1al. Using three methods of identification, SDS-PAGE, RP-HPLC and PCR marker analysis, as well as pedigree information, we have traced the distribution and source of this allele from a Uruguayan landrace, Americano 44D, in the mid-nineteenth century. Results are supported by knowledge of the movement of wheat lines with migrants. All cultivars possessing the Glu-B1al allele can be identified by the following attributes: (1) the elution of the By sub-unit peak before the Dx sub-unit peak by RP-HPLC, (2) high expression levels of Bx7 (>39% Mol% Bx), (3) a 43 bp insertion in the matrix-attachment region (MAR) upstream of the gene promoter relative to Bx7 and an 18 bp nucleotide duplication in the coding region of the gene. Evidence is presented indicating that these 18 and 43 bp sequence insertions are not causal for the high expression levels of Bx7 as they were also found to be present in a small number of hexaploid species, including Chinese Spring, and species expressing Glu-B1ak and Glu-B1a alleles. In addition, these sequence inserts were found in different isolates of the tetraploid wheat, T. turgidum, indicating that these insertion/deletion events occurred prior to hexaploidization.

  12. Research Article Allelic Variation of High Molecular Weight Glutenin ...

    Indian Academy of Sciences (India)

    Sky123.Org

    ... food crops and can be processed into multiple types of food products (e.g., ... Bread-making quality of wheat is determined to a large extent by the properties of the ... In Hebei Province, the study of genetic improvement of wheat quality has ...

  13. Influence of low-molecular-weight glutenin subunit haplotypes on dough rheology and baking quality in elite common wheat varieties

    Science.gov (United States)

    The low molecular weight glutenin subunits (LMW-GSs) are a class of wheat seed storage proteins directly involved in the formation of gluten. Depending on the first amino acid residue of the mature proteins, the LMW-GSs are divided into methionine, serine or isoleucine type. These proteins are encod...

  14. Effects of HMW-GS Ax1 or Dx2 absence on the glutenin polymerization and gluten micro structure of wheat (Triticum aestivum L.).

    Science.gov (United States)

    Gao, Xin; Liu, Tianhong; Ding, Mengyun; Wang, Jun; Li, Chunlian; Wang, Zhonghua; Li, Xuejun

    2018-02-01

    Wheat (Triticum aestivum L.) dough strength and extensibility are mainly determined by the polymerization of glutenin. The number of high-molecular-weight glutenin subunits (HMW-GS) differs in various wheat varieties due to the silencing of some genes. The effects of Ax1 or Dx2 subunit absence on glutenin polymerization, dough mixing properties and gluten micro structure were investigated with two groups of near-isogenic lines. The results showed that Ax1 or Dx2 absence decreased the accumulation rate of glutenin polymers and thus delayed the rapid increase period for glutenin polymerization by at least ten days, which led to lower percentage of polymeric protein in mature grain. Ax1 or Dx2 absence significantly decreased the dough development time and dough stability, but increased the uniformity of micro structure. Lacunarity, derived from quantitative analysis of gluten network, is suggested as a new indicator for wheat quality. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Evaluation of Pakistani wheat germplasm for bread quality based on allelic variation in HMW glutenin subunits

    Energy Technology Data Exchange (ETDEWEB)

    Tabasum, A; Iqbal, N; Hameed, A; Arshad, R [Nuclear Institute for Agriculture and Biology, Faisalabad (Pakistan)

    2011-06-15

    Seventy six Pakistani wheat genotypes including land races were investigated for Bread quality (BQ) based on allelic variation in HMW glutenin subunits at the Glu-1 loci through SDS- polyacrylamide gel electropherosis. Twenty five different allelic combinations were detected with a total of 14 Glu-1 loci. Highest polymorphism was revealed by Glu-B locus and some single/ rare sub units were also screened out. The frequencies of dominant subunits were 50% for 2*, 42.11% for subunit pair 17+18 and 48.68% for 5+10 and 2+12 respectively. The quality scores displayed a range from 4 to 10, however generally good quality score of eight was more frequent (39. 47%). The highest quality scores of 10 and 9 were observed in 22.36% and 19.74% of genotypes respectively. The UPGMA analysis grouped genotypes into three major with two additional sub clusters for each. The cluster 'a' 'b' and 'C' were separated at 73% genetic distance which was further differentiated at a genetic distance of 50% into their sub clusters. Pakistani wheat varieties/land races exhibited large variation in term of HMW-GS. The generated information will lead to the pyrimiding of sub units for high BQ through mission oriented marker assisted breeding programmes for quality improvement of wheat. (author)

  16. Evaluation of Pakistani wheat germplasm for bread quality based on allelic variation in HMW glutenin subunits

    International Nuclear Information System (INIS)

    Tabasum, A.; Iqbal, N.; Hameed, A.; Arshad, R.

    2011-01-01

    Seventy six Pakistani wheat genotypes including land races were investigated for Bread quality (BQ) based on allelic variation in HMW glutenin subunits at the Glu-1 loci through SDS- polyacrylamide gel electropherosis. Twenty five different allelic combinations were detected with a total of 14 Glu-1 loci. Highest polymorphism was revealed by Glu-B locus and some single/ rare sub units were also screened out. The frequencies of dominant subunits were 50% for 2*, 42.11% for subunit pair 17+18 and 48.68% for 5+10 and 2+12 respectively. The quality scores displayed a range from 4 to 10, however generally good quality score of eight was more frequent (39. 47%). The highest quality scores of 10 and 9 were observed in 22.36% and 19.74% of genotypes respectively. The UPGMA analysis grouped genotypes into three major with two additional sub clusters for each. The cluster 'a' 'b' and 'C' were separated at 73% genetic distance which was further differentiated at a genetic distance of 50% into their sub clusters. Pakistani wheat varieties/land races exhibited large variation in term of HMW-GS. The generated information will lead to the pyrimiding of sub units for high BQ through mission oriented marker assisted breeding programmes for quality improvement of wheat. (author)

  17. High Molecular Weight Melanoidins from Coffee Brew

    NARCIS (Netherlands)

    Bekedam, E.K.; Schols, H.A.; Boekel, van T.; Smit, G.

    2006-01-01

    The composition of high molecular weight (HMw) coffee melanoidin populations, obtained after ethanol precipitation, was studied. The specific extinction coefficient (Kmix) at 280, 325, 405 nm, sugar composition, phenolic group content, nitrogen content, amino acid composition, and non-protein

  18. Chemometric Analysis of High Molecular Mass Glutenin Subunits and Image Data of Bread Crumb Structure from Croatian Wheat Cultivars

    Directory of Open Access Journals (Sweden)

    Zorica Jurković

    2002-01-01

    Full Text Available The aim of this work is to investigate functional relationships among wheat properties, high molecular mass (weight (HMW glutenin subunits and bread quality produced from eleven Croatian wheat cultivars by chemometric analysis. HMW glutenin subunits were fractionated by sodium dodecylsulfate polyacrylamid gel electrophoresis (SDS-PAGE and subsequently analysed by scanning densitometry in order to quantify HMW glutenin fractions. Wheat properties are characterised by four variables: protein content, sedimentation value, wet gluten and gluten index. Bread quality is assessed by the standard measurement of loaf volume, and visual quality of bread slice is quantified by 8 parameters by the use of computer image analysis. The data matrix with 21 columns (measured variables and 11 rows (cultivars is analysed for determination of number of latent variables. It was found that the first two latent variables account for 92, 85 and 87 % of variance of wheat quality properties, HMW glutenin fractions, and the bread quality parameters, respectively. Classification and functional relationships are discussed from the case data (cultivars and variable projections to the planes of the first two latent variables. Between Glu-D1y proportion and the bread quality parameters (standard parameter loaf volume and bread crumb cell area fraction determined by image analysis the strongest positive correlations are found r = 0.651 and r = 0.885, respectively. Between Glu-B1x proportion and the bread quality parameters the strongest negative correlations are found r =-0.535 and r = –0.841, respectively. The results are discussed in view of possible development of new and improvement of existing wheat cultivars and optimisation of bread production.

  19. Sintering of ultra high molecular weight polyethylene

    Indian Academy of Sciences (India)

    Abstract. Ultra high molecular weight polyethylene (UHMWPE) is a high performance polymer having low coefficient of friction, good abrasion resistance, good chemical ... In this study, we report our results on compaction and sintering behaviour of two grades of UHMWPE with reference to the powder morphology, sintering ...

  20. Definition of the low molecular weight glutenin subunit gene family members in a set of standard bread wheat (Triticum aestivum L.) varieties

    Science.gov (United States)

    Low-molecular-weight glutenin subunits (LMW-GS) are a class of seed storage proteins that play a major role in the determination of the viscoelastic properties of wheat dough. Most of the LMW-GSs are encoded by a multi-gene family located on the short arms of the homoeologous group 1 chromosomes, at...

  1. Variation in concentrations of high-molecular-weight glutenin subunits and macropolymers in wheat grains of a recombinant inbred lines population and in two contrasting eco-sites in China

    DEFF Research Database (Denmark)

    Li, Xiangnan; Cai, Jian; Liu, Fulai

    2012-01-01

    to be significantly correlated with the GMP concentration. CONCLUSION: Concentrations of HMW-GS andGMPare largely affected by the eco-sites and the composition ofHMW-GS, whilst not by the presence of 6 VS·6AL chromosome segment translocation. The 6 VS·6AL translocation is suggested as potential donor for breeding...... wheat cultivars for high resistence to powdery mildew and yellow rust with less risk of undesirable effects on grain quality. c 2012 Society of Chemical Industry Supporting information may be found in the online version of this article....

  2. Nasal Delivery of High Molecular Weight Drugs

    Directory of Open Access Journals (Sweden)

    Erdal Cevher

    2009-09-01

    Full Text Available Nasal drug delivery may be used for either local or systemic effects. Low molecular weight drugs with are rapidly absorbed through nasal mucosa. The main reasons for this are the high permeability, fairly wide absorption area, porous and thin endothelial basement membrane of the nasal epithelium. Despite the many advantages of the nasal route, limitations such as the high molecular weight (HMW of drugs may impede drug absorption through the nasal mucosa. Recent studies have focused particularly on the nasal application of HMW therapeutic agents such as peptide-protein drugs and vaccines intended for systemic effects. Due to their hydrophilic structure, the nasal bioavailability of peptide and protein drugs is normally less than 1%. Besides their weak mucosal membrane permeability and enzymatic degradation in nasal mucosa, these drugs are rapidly cleared from the nasal cavity after administration because of mucociliary clearance. There are many approaches for increasing the residence time of drug formulations in the nasal cavity resulting in enhanced drug absorption. In this review article, nasal route and transport mechanisms across the nasal mucosa will be briefly presented. In the second part, current studies regarding the nasal application of macromolecular drugs and vaccines with nanoand micro-particulate carrier systems will be summarised.

  3. Cloning and Characterization of Low-Molecular-Weight Glutenin Subunit Alleles from Chinese Wheat Landraces (Triticum aestivum L.

    Directory of Open Access Journals (Sweden)

    Hongqi Si

    2014-01-01

    Full Text Available Low-molecular-weight glutenin subunits (LMW-GS are of great importance in processing quality and participate in the formation of polymers in wheat. In this study, eight new LMW-GS alleles were isolated from Chinese wheat landraces (Triticum aestivum L. and designated as Glu-A3-1a, Glu-A3-1b, Glu-B3-1a, Glu-B3-1b, Glu-B3-1c, Glu-D3-1a, Glu-D3-1b, and Glu-D3-1c, which were located at the Glu-A3, Glu-B3, and Glu-D3 loci, respectively. Based on the proteins encoded, the number of deduced amino acids of Glu-B3 alleles was approximately 50 more than those of Glu-A3 and Glu-D3 alleles. The first cysteine of Glu-A3 and Glu-D3 alleles was located at the N-terminal domain, while that of Glu-B3 alleles was found in the repetitive domain, which may lead to the different functioning in forming disulfide bonds. All the eight genes were LMW-m types and the new allele of Glu-B3-1a which had nine cysteine residues may be the desirable LMW-GS gene for improving bread-making quality.

  4. Isolation and Sequence Analysis of HMW Glutenin Subunit 1Dy10.1 Ecoding Gene from Xinjiang Wheat (Triticum petropavlovskyi Udacz.et Migusch)

    Institute of Scientific and Technical Information of China (English)

    JIANG Qian-tao; WEI Yu-ming; WANG Ji-rui; YAN Ze-hong; ZHENG You-liang

    2006-01-01

    A novel HMW glutenin subunit gene 1Dy10.1 was isolated and characterized from Xinjiang wheat (Triticum petropavlovskyi. Udacz. et Migusch) accession Daomai 2. The complete open reading frame (ORF) of 1Dy10.1 was 1965 bp, encoding 655 amino acids. The numbers and distribution of cysteines in 1Dy10.1 were similar to those of 1Dy10 and other y-type subunits. In the N-terminal of 1Dy10.1, an amino acid was changed from L (leucine) to P (proline) at position 55. The repetitive domain of 1Dy10.1 differed from those of known HMW subunits by substitutions, insertions or/and deletions involving single or more amino acid residues. In the repetitive domain of subunit 1Dy10.1, the deletion of tripeptide GQQ in the consensus unit PGQGQQ resulted in the appearance of the motif PGQ that have not been observed in other known y-type HMW subunits. In comparison with the subunit 1Dy12, a deletion of dipeptide GQ, which occurred in subunit 1Dy10, was also observed in subunit 1Dy10.1. The cloned 1Dyl0.1 gene had been successfully expressed in Escherichia coli, and the expressed protein had the identical mobility with the endogenous subunit 1Dyl0.1 from seed.

  5. High Molecular Weight Polymers in the New Chemicals Program

    Science.gov (United States)

    There are three categories or types of High Molecular Weight (HMW, 10,000 daltons) polymers typically reviewed by the New Chemicals Program: Soluble, insoluble, and water absorbing. Each of the three types are treated differently.

  6. Development of radiation-resisting high molecular-weight materials

    International Nuclear Information System (INIS)

    Nakagawa, Tsutomu

    1976-01-01

    The excellent radiation-resisting polyvinyl chloride developed at the opportunity of the research on the relationships between the protection of living body and the polymer-technological protection from radiation is reviewed. The report is divided into four main parts, namely 1) the change in the molecular arrangement of market-available, high molecular-weight materials by gamma-ray irradiation, 2) the protection of high molecular-weight materials from radiation, 3) the relationships between the biological radiation-protective substances and the change to radiation-resisting property of synthesized high molecular-weight substances, and 4) the development of the radiation-resisting high molecular-weight materials as metal-collecting agents. Attention is paid to the polyvinyl chloride having N-methyl-dithio-carbamate radical (PMD), synthesized by the author et. al., that has excellent radiation-resisting property. PMD has some possibility to form thiol- and amino-radicals necessary to protect living things from radiation. It is believed that the protection effects of N-methyl-dithio-carbamate radical are caused by the relatively stable S radical produced by the energy transfer. PMD film is suitable for the irradiation of foods, because it hardly changes the permeability of oxygen and carbon dioxide. PMD produces mercaptide or chelate. A new metal-collecting agent (PSDC) having reactivity with the metallic ions with radiation-resisting property was developed, which is derived from polyvinyl chloride and sodium N-methyl-N-carboxy-methyl-dithio-carbamate. (Iwakiri, K.)

  7. HMW glutenin variation and rye chromatine presence in wheat genome

    Directory of Open Access Journals (Sweden)

    Obreht Dragana R.

    2003-01-01

    Full Text Available For estimation of wheat end-product quality during wheat breeding programs composition of high-molecular-weight glutenin subunits (HMW GS and the presence of 1BL/1RS translations serve as markers due to their profound effects on dough elasticity and viscous properties. Ninety-three wheat genotypes from Institute of Field and Vegetable Crops in Novi Sad have been analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE in order to determine their HMW GS composition and 1BL/1RS translocation presence. Eleven alleles were found at the Glu-1 loci. Subunits 1 and 2*and the null allele N were determined at the Glu-A1 locus. Subunitis 7, 7+9, 7+8, 6+8, 20 and 21 were found at the Glu-B1 locus, subunits 2+12 and 5+10 at the Glu-D1 locus. The 1BL/1RS translocation was discovered in 28 cultivars, although three of them were heterogeneous.

  8. Solvent extraction of cerium (III) with high molecular weight amines

    International Nuclear Information System (INIS)

    Chatterjee, A.; Basu, S.

    1992-01-01

    The use of high molecular weight amines in the extraction of cerium (III) as EDTA complex from neutral aqueous medium is reported. The extraction condition was optimised from the study of effects of several variables like concentration of amine and EDTA pH nature of diluents etc. The method has been applied for the determination of cerium in few mineral samples. (author). 7 refs., 5 tabs

  9. High molecular weight polysaccharide that binds and inhibits virus

    Science.gov (United States)

    Konowalchuk, Thomas W

    2014-01-14

    This invention provides a high molecular weight polysaccharide capable of binding to and inhibiting virus and related pharmaceutical formulations and methods on inhibiting viral infectivity and/or pathogenicity, as well as immunogenic compositions. The invention further methods of inhibiting the growth of cancer cells and of ameliorating a symptom of aging. Additionally, the invention provides methods of detecting and/or quantifying and/or isolating viruses.

  10. High molecular weight polysaccharide that binds and inhibits virus

    Energy Technology Data Exchange (ETDEWEB)

    Konowalchuk, Thomas W.; Konowalchuk, Jack

    2017-07-18

    This invention provides a high molecular weight polysaccharide capable of binding to and inhibiting virus and related pharmaceutical formulations and methods of inhibiting viral infectivity and/or pathogenicity, as well as immunogenic compositions. The invention further includes methods of inhibiting the growth of cancer cells and of ameliorating a symptom of aging. Additionally, the invention provides methods of detecting and/or quantifying and/or isolating viruses.

  11. Relationship between the dough quality and content of specific glutenin proteins in wheat mill streams, and its application to making flour suitable for instant Chinese noodles.

    Science.gov (United States)

    Yahata, Eriko; Maruyama-Funatsuki, Wakako; Nishio, Zenta; Yamamoto, Yoshihiko; Hanaoka, Akihiro; Sugiyama, Hisashi; Tanida, Masatoshi; Saruyama, Haruo

    2006-04-01

    The content of specific proteins such as high-molecular-weight glutenin subunits HMW-GS 5+10 and low-molecular-weight glutenin subunits LMW-GS KS2 in wheat mill streams of extra-strong Kachikei 33 wheat was quantified by SDS-PAGE and 2D-PAGE. The mill streams showed varied quantities of HMW-GS 5+10 (0.077 to 2.007 mg/g of mill stream), LMW-GS KS2 (0.018 to 0.586 mg/g of mill stream) and total protein (9.42% to 18.98%). The contents of these specific proteins in the mill streams were significantly correlated with the SDS sedimentation volume and the mixing properties, which are respective indices of specific loaf volume and dough strength. The contents of these specific glutenin proteins in the mill streams were therefore found to be significantly important for improving the dough quality suitable for bread and Chinese noodles. Accordingly, we present here the application of this information to the development of an effective method for producing mill streams with high quality and yield that are suitable for instant Chinese noodles.

  12. Steroidogenic activity of high molecular weight forms of ACTH

    International Nuclear Information System (INIS)

    Gasson, J.C.

    1979-01-01

    The relative steroidogenic potencies of high molecular weight forms of adrenocorticotropic hormone (ACTH) were investigated using in vitro bioassays. In order to prepare pools of separated pro-ACTH/endorphin, ACTH biosynthetic intermediate and glycosylated ACTH (1-39), the protein present in serum-free tissue culture medium obtained from cultured AtT-20/D-16v mouse pituitary tumor cells was concentrated and fractionated by gel filtration. Based on sodium dodecyl sulfate polyacrylamide gel electrophoresis, over 97% of the immunoactive ACTH in each pool had the appropriate molecular weight. Suspensions of isolated rat and guinea pig adrenal cortical cells were prepared by enzymatic dissociation and mechanical dispersion. Cells were incubated in complete tissue culture medium overnight then used in a 2 hour steroid production assay. Synthetic hACTH(1-39) was used as a bioassay and immunoassay standard. The amounts of pro-ACTH/endorphin, ACTH biosynthetic intermediate and glycosylated ACTH(1-39) bioassayed were estimated by ACTH(17-24) radioimmunoassay. All three high molecular weight forms of ACTH were capable of stimulating the same maximal level of steroidogenesis, by both isolated rat and guinea pig adrenal cells, as hACTH(1-39). Glycosylated ACTH(1-39) was equipotent with hACTH(1-39); pro-ACTH/endorphin and ACTH biosynthetic intermediate were two orders of magnitude less potent than hACTH(1-39) in both bioassay systems

  13. Removal of high-molecular weight polycyclic aromatic hydrocarbons

    Directory of Open Access Journals (Sweden)

    Ulrich Vasconcelos

    2011-01-01

    Full Text Available Alternatives for the removal of high-molecular weight polycyclic aromatic hydrocarbons (HWM-PAH from soil were tested by adding fertilizer or glycerol, as well as the combination of both. Experiments were carried out for 60 days in reactors containing a HWM-PAH-contaminated soil (8030 μg kg-1, accompanied by pH monitoring, humidity control and quantification of total heterotrophic bacteria and total fungus. Fertilizer addition removed 41.6% of HWM-PAH. Fertilizer and glycerol in combination removed 46.2%. When glycerol was added individually, degradation reached 50.4%. Glycerol also promoted the increase of degradation rate during the first 30 days suggesting the HMW-PAH removal occurred through cometabolic pathways.

  14. Development of a new marker system for identifying the complex members of the low-molecular-weight glutenin subunit gene family in bread wheat (Triticum aestivum L.).

    Science.gov (United States)

    Zhang, Xiaofei; Liu, Dongcheng; Yang, Wenlong; Liu, Kunfan; Sun, Jiazhu; Guo, Xiaoli; Li, Yiwen; Wang, Daowen; Ling, Hongqing; Zhang, Aimin

    2011-05-01

    Low-molecular-weight glutenin subunits (LMW-GSs) play an important role in determining the bread-making quality of bread wheat. However, LMW-GSs display high polymorphic protein complexes encoded by multiple genes, and elucidating the complex LMW-GS gene family in bread wheat remains challenging. In the present study, using conventional polymerase chain reaction (PCR) with conserved primers and high-resolution capillary electrophoresis, we developed a new molecular marker system for identifying LMW-GS gene family members. Based on sequence alignment of 13 LMW-GS genes previously identified in the Chinese bread wheat variety Xiaoyan 54 and other genes available in GenBank, PCR primers were developed and assigned to conserved sequences spanning the length polymorphism regions of LMW-GS genes. After PCR amplification, 17 DNA fragments in Xiaoyan 54 were detected using capillary electrophoresis. In total, 13 fragments were identical to previously identified LMW-GS genes, and the other 4 were derived from unique LMW-GS genes by sequencing. This marker system was also used to identify LMW-GS genes in Chinese Spring and its group 1 nulli-tetrasomic lines. Among the 17 detected DNA fragments, 4 were located on chromosome 1A, 5 on 1B, and 8 on 1D. The results suggest that this marker system is useful for large-scale identification of LMW-GS genes in bread wheat varieties, and for the selection of desirable LMW-GS genes to improve the bread-making quality in wheat molecular breeding programmes.

  15. PCR-based isolation and identification of full-length low-molecular-weight glutenin subunit genes in bread wheat (Triticum aestivum L.).

    Science.gov (United States)

    Zhang, Xiaofei; Liu, Dongcheng; Jiang, Wei; Guo, Xiaoli; Yang, Wenlong; Sun, Jiazhu; Ling, Hongqing; Zhang, Aimin

    2011-12-01

    Low-molecular-weight glutenin subunits (LMW-GSs) are encoded by a multi-gene family and are essential for determining the quality of wheat flour products, such as bread and noodles. However, the exact role or contribution of individual LMW-GS genes to wheat quality remains unclear. This is, at least in part, due to the difficulty in characterizing complete sequences of all LMW-GS gene family members in bread wheat. To identify full-length LMW-GS genes, a polymerase chain reaction (PCR)-based method was established, consisting of newly designed conserved primers and the previously developed LMW-GS gene molecular marker system. Using the PCR-based method, 17 LMW-GS genes were identified and characterized in Xiaoyan 54, of which 12 contained full-length sequences. Sequence alignments showed that 13 LMW-GS genes were identical to those found in Xiaoyan 54 using the genomic DNA library screening, and the other four full-length LMW-GS genes were first isolated from Xiaoyan 54. In Chinese Spring, 16 unique LMW-GS genes were isolated, and 13 of them contained full-length coding sequences. Additionally, 16 and 17 LMW-GS genes in Dongnong 101 and Lvhan 328 (chosen from the micro-core collections of Chinese germplasm), respectively, were also identified. Sequence alignments revealed that at least 15 LMW-GS genes were common in the four wheat varieties, and allelic variants of each gene shared high sequence identities (>95%) but exhibited length polymorphism in repetitive regions. This study provides a PCR-based method for efficiently identifying LMW-GS genes in bread wheat, which will improve the characterization of complex members of the LMW-GS gene family and facilitate the understanding of their contributions to wheat quality.

  16. Properties of crosslinked ultra-high-molecular-weight polyethylene.

    Science.gov (United States)

    Lewis, G

    2001-02-01

    Substantially reducing the rate of generation of wear particles at the surfaces of ultra-high-molecular-weight polyethylene (UHMWPE) orthopedic implant bearing components, in vivo, is widely regarded as one of the most formidable challenges in modern arthroplasty. In the light of this, much research attention has been paid to the myriad of endogenous and exogenous factors that have been postulated to affect this wear rate, one such factor being the polymer itself. In recent years, there has been a resurgence of interest in crosslinking the polymer as a way of improving its properties that are considered relevant to its use for fabricating bearing components. Such properties include wear resistance, fatigue life, and fatigue crack propagation rate. Although a large volume of literature exists on the topic on the impact of crosslinking on the properties of UHMWPE, no critical appraisal of this literature has been published. This is one of the goals of the present article, which emphasizes three aspects. The first is the trade-off between improvement in wear resistance and depreciation in other mechanical and physical properties. The second aspect is the presentation of a method of estimating the optimal value of a crosslinking process variable (such as dose in radiation-induced crosslinking) that takes into account this trade-off. The third aspect is the description of a collection of under- and unexplored research areas in the field of crosslinked UHMWPE, such as the role of starting resin on the properties of the crosslinked polymer, and the in vitro evaluation of the wear rate of crosslinked tibial inserts and other bearing components that, in vivo, are subjected to nearly unidirectional motion.

  17. Glomerular sieving of high molecular weight proteins in proteinuric rats

    International Nuclear Information System (INIS)

    Bertolatus, J.A.; Abuyousef, M.; Hunsicker, L.G.

    1987-01-01

    To characterize the permeability of the glomerular capillary wall to high molecular weight proteins in normal and proteinuric rats, we determined the glomerular sieving coefficients (GSC) of radioiodinated marker proteins of known size and charge by means of a paired label, tissue accumulation method previously validated in this laboratory. In one group of rats (Series A) the GSCs of 125 I-anionic IgG (aIgG-molecular weight [mol wt] 150,000, pI 4.9) and 131 I-neutral IgG (nIgG-pI 7.4 to 7.6) were measured simultaneously. In Series B, the GSC of a second anionic marker, 131 I-human ceruloplasmin (Crp-mol wt 137,000, pI 4.9) was compared to that of 125 I-nIgG. As in the previous report, the labeled proteins were not degraded or deiodinated during the 20 minute clearance period for GSC determination. Within Series A and B, three subgroups of rats were studied: control saline-infused rats, rats made acutely proteinuric by infusion of the polycation hexadimethrine (HDM), and rats with chronic doxorubicin (Adriamycin-Adria) nephrosis. In the control rats, GSCs for the anionic markers aIgG (Series A) or Crp (Series B) were significantly greater than that of nIgG (both series). These large proteins crossed the filtration barrier by a different pathway from that available to smaller neutral molecules the size of albumin, which in our previous study had a much higher GSC than a native, anionic albumin marker. In a third group of control rats only (Series C), the GSCs of native anionic bovine albumin (BSA) and nIgG were compared directly. The GSC of BSA (0.0029) was only slightly larger than the GSC of nIgG (0.0025), indicating that most of the native albumin crosses the glomerular capillary wall via a nonselective pathway similar to that available to nIgG. The results in the control groups are compatible with recently-described heteroporous models of glomerular size selectivity

  18. Identification of coeliac disease triggering glutenin peptides in adults.

    Science.gov (United States)

    Donnelly, Suzanne C; Šuligoj, Tanja; Ellis, H Julia; Ciclitira, Paul J

    2016-07-01

    Coeliac disease affects approximately 1% of Northern American and European populations. It is caused by an inappropriate immune response to dietary gluten. Gluten comprises of two major protein fractions: gliadins and glutenins. Glutenins have recently been found to be toxic to coeliac individuals. Proliferation assays suggest in some but not all paediatric coeliac individuals there may be immunological stimulation with high molecular weight (HMW) glutenins. Less evidence pertains to low molecular weight (LMW) glutenins. The aim is to assess adaptive, T-cell driven, and innate immune response in adult coeliac individuals towards HMW glutenin peptide, glut04, and LMW glutenin peptide, glt156. Coeliac patients were recruited attending endoscopy for routine monitoring. Adaptive immune response towards glut04 and glt156 was measured by proliferation assays and measurement of interferon-γ secretion in 28 T-cell lines. The innate immune response was assessed by measurement of enterocyte cell height (ECH) in coeliac small intestinal biopsies following overnight incubation in organ culture chambers in a further nine individuals. There were 3/28 and 2/28 positive proliferation results using gluten-sensitive T-cells with glut04 and glt156, respectively. All coeliac biopsies tested in organ culture chambers demonstrated clear reduction in ECH with peptic-tryptic digest of whole industrial gluten, glut04 and glt156 when compared to negative control ovalbumin (p < 0.005). Three individuals had both T-cell and organ culture study data. Their proliferation assays showed no stimulation of the T-cells. This study demonstrates glutenin epitopes glut04 and glt156, while minor T-cell epitopes, are important in their ability to trigger the innate immune response.

  19. CHARACTERIZATION OF GLIADIN AND HMW GLUTENIN PROTEIN COMPOSITION IN COLOURED WHEAT (TRITICUM AESTIVUM L. VARIETIES

    Directory of Open Access Journals (Sweden)

    Valéria Šudyová

    2011-12-01

    Full Text Available Wheat is one of the most important grains in our daily diet. Coloured wheat contains natural anthocyanin compounds. Bioactive compounds in wheat have attracted increasingly more interest from breeders because of their benefits. It is important to fully understand protein properties of red, blue, purple, and yellow-coloured wheat in order to predict their potential uses for culturing new varieties. All 21 accessions originating from different geographical areas of world were evaluated for high molecular weight glutenin subunit (HMW-GS and T1BL.1RS wheat-rye translocation using SDS-PAGE and A-PAGE. The data indicated the prevalence of the allele 1 (36%, allele 0 (30% and allele 2* (34% at the Glu-1A and five alleles, namely 7+8 (36%, 7+9 (29%, 20 (21%, 7 (12% and 17+18 (2% represented the Glu-1B. Existence of 2 alleles at the locus Glu-1D was revealed, in fact 21% of them showed the subunit pairs Glu-1D 5+10 correlated with good bread making properties. Protein subunit Glu-1A1 and Glu-1A2* were correlated positively with improved dough strength as compared to subunit null. On the chromosome Glu-1B subunit 17+18 and 7+8 were associated with slightly stronger gluten type than 7+9, whereas subunit 20 and 7 were associated with weak gluten properties. On the basis of electrophoretic separation of gliadin fraction it was found that only one genotype contained T1BL.1RS wheat-rye translocation. The Glu-1 quality score ranged from 4 to 10. Suitable accessions can be used for the crossing programs to improve colour and good technological quality of bread wheat.  doi:10.5219/161

  20. Disassembled DJ-1 high molecular weight complex in cortex mitochondria from Parkinson's disease patients

    Directory of Open Access Journals (Sweden)

    Adler Charles

    2009-07-01

    Full Text Available Abstract Correction to Nural H, He P, Beach T, Sue L, Xia W, Shen Y. Disassembled DJ-1 high molecular weight complex in cortex mitochondria from Parkinson's disease patients Molecular Neurodegeneration 2009, 4:23.

  1. The effects of nitrogen nutrition and glutenin composition on the gluten quality in wheat genotypes

    Directory of Open Access Journals (Sweden)

    NIKOLA HRISTOV

    2010-03-01

    Full Text Available The effect of nitrogen nutrition treatments on the gluten content and some quality parameters of eight winter wheat cultivars has been studied. Six different nitrogen rates were applied (0, 60, 90, 120, 150 and 180 kg N ha-1 to wheat cultivars chosen according to the structure of their high molecular weight glutenin subunits (HMW-GS at the Glu-D1 locus. Four genotypes with HMW-GS 2 + 12 and another four with HMW-GS 5 + 10 were used in the study. The analysis of gluten quality involved the wet gluten content and rheological properties determined by the sensory and instrumental methods (“Instron 4301”. It was determined that in all the cultivars the wet gluten content increased significantly (P < 0.05 in parallel with N rate increase. The cultivars reacted differently regarding their wet gluten rheological properties. Libellula, a cultivar with poor bread making quality (HMW-GS 2 + 12, did not react to different N rates. Sremica, a cultivar with excellent bread making quality (HMW-GS 5 + 10, reduced its gluten quality as the N rate increased. The values obtained by the instrumental method “Instron 4301” at 90% wet gluten compression varied widely (from 0.002 to 0.041 kN. The increase of N fertilizer rate was significantly positively correlated (r2 = 0.811 with the wet gluten content and strength in the cultivars with HMW-GS 5+10.

  2. Loss of high-molecular-weight cytokeratin antigenicity in prostate tissue obtained by transurethral resections

    DEFF Research Database (Denmark)

    Multhaupt, H A; Fessler, J N; Warhol, M J

    2000-01-01

    could be restored in these specimens by antigen retrieval in a low pH citrate buffer using a microwave heat technique. Keratin staining in needle biopsies and total prostatectomies was unaffected. CONCLUSION: In summary, our results indicate the technique of transurethral resection results in a specific......OBJECTIVE: Staining of prostatic basal cells for the expression of high-molecular-weight cytokeratin has been suggested as a way of distinguishing benign from malignant prostate glands. We evaluated the utility of high-molecular-weight cytokeratin in the diagnosis of malignancy in prostate...... specimens obtained in various ways. DESIGN: Prostate tissues obtained from needle biopsies, transurethral resections, and total prostatectomies were immunostained with monoclonal antibody 34betaE12, an antibody directed against high-molecular-weight cytokeratins. RESULTS: Antiserum to high...

  3. Thermal characterization of Ag and Ag + N ion implanted ultra-high molecular weight polyethylene (UHMWPE)

    Science.gov (United States)

    Sokullu Urkac, E.; Oztarhan, A.; Tihminlioglu, F.; Kaya, N.; Ila, D.; Muntele, C.; Budak, S.; Oks, E.; Nikolaev, A.; Ezdesir, A.; Tek, Z.

    2007-08-01

    Most of total hip joints are composed of ultra-high molecular weight polyethylene (UHMWPE). However, as ultra-high molecular weight polyethylene is too stable in a body, wear debris may accumulate and cause biological response such as bone absorption and loosening of prosthesis. In this study, ultra-high molecular weight polyethylene samples were Ag and Ag + N hybrid ion implanted by using MEVVA ion implantation technique to improve its surface properties. Samples were implanted with a fluence of 1017 ion/cm2 and extraction voltage of 30 kV. Implanted and unimplanted samples were investigated by thermo-gravimetry analysis (TGA), differential scanning calorimetry (DSC), X-ray diffraction (XRD) analysis, scanning electron microscopy (SEM), optical microscopy (OM) and contact Angle measurement. Thermal characterization results showed that the ion bombardment induced an increase in the % crystallinity, onset and termination degradation temperatures of UHMWPE.

  4. Thermal characterization of Ag and Ag + N ion implanted ultra-high molecular weight polyethylene (UHMWPE)

    Energy Technology Data Exchange (ETDEWEB)

    Sokullu Urkac, E. [Department of Materials Science, Izmir High Technology Institute, Gulbahcekoyu Urla, Izmir (Turkey)]. E-mail: emelsu@gmail.com; Oztarhan, A. [Bioengineering Department, Ege University, Bornova, Izmir 35100 (Turkey); Tihminlioglu, F. [Department of Chemical Engineering, Izmir High Technology Institute, Gulbahcekoyu Urla, Izmir (Turkey); Kaya, N. [Bioengineering Department, Ege University, Bornova, Izmir 35100 (Turkey); Ila, D. [Center for Irradiation of Materials, Alabama A and M University, Normal AL 35762 (United States); Muntele, C. [Center for Irradiation of Materials, Alabama A and M University, Normal AL 35762 (United States); Budak, S. [Center for Irradiation of Materials, Alabama A and M University, Normal AL 35762 (United States); Oks, E. [H C Electronics Institute, Tomsk (Russian Federation); Nikolaev, A. [H C Electronics Institute, Tomsk (Russian Federation); Ezdesir, A. [R and D Department, PETKIM Holding A.S., Aliaga, Izmir 35801 (Turkey); Tek, Z. [Department of Physics, Celal Bayar University, Manisa (Turkey)

    2007-08-15

    Most of total hip joints are composed of ultra-high molecular weight polyethylene (UHMWPE ). However, as ultra-high molecular weight polyethylene is too stable in a body, wear debris may accumulate and cause biological response such as bone absorption and loosening of prosthesis. In this study, ultra-high molecular weight polyethylene samples were Ag and Ag + N hybrid ion implanted by using MEVVA ion implantation technique to improve its surface properties. Samples were implanted with a fluence of 10{sup 17} ion/cm{sup 2} and extraction voltage of 30 kV. Implanted and unimplanted samples were investigated by thermo-gravimetry analysis (TGA), differential scanning calorimetry (DSC), X-ray diffraction (XRD) analysis, scanning electron microscopy (SEM), optical microscopy (OM) and contact Angle measurement. Thermal characterization results showed that the ion bombardment induced an increase in the % crystallinity, onset and termination degradation temperatures of UHMWPE.

  5. Reversible, high molecular weight palladium and platinum coordination polymers based on phosphorus ligands

    NARCIS (Netherlands)

    Paulusse, J.M.J.; Huijbers, J.P.J.; Sijbesma, R.P.

    2005-01-01

    A general strategy for the preparation and characterization of high molecular weight coordination polymers based on bifunctional phosphorus ligands and palladium or platinum dichloride is described. Metal-to-ligand stoichiometry is of key importance for the formation of linear coordination polymers

  6. Reversible, High Molecular Weight Palladium and Platinum Coordination Polymers Based on Phosphorus Ligands

    NARCIS (Netherlands)

    Paulusse, Jos Marie Johannes; Huijbers, Jeroen P.J.; Sijbesma, Rint P.

    2005-01-01

    A general strategy for the preparation and characterization of high molecular weight coordination polymers based on bifunctional phosphorus ligands and palladium or platinum dichloride is described. Metal-to-ligand stoichiometry is of key importance for the formation of linear coordination polymers

  7. The adhesive properties of chlorinated ultra-high molecular weight polyethylene

    NARCIS (Netherlands)

    Menting, H.N.A.M.; Voets, P.E.L.; Lemstra, P.J.

    1995-01-01

    Ultra-high molecular weight polyethylene (UHMW-PE) is well known for its abrasion and chemical resistance. Recently we developed a new application for UHMW-PE as a liner in elastomeric hoses. It was found that the adhesion between UHMW-PE and elastomers such as ethylene-propylene-diene monomer

  8. Extension induced phase separation and crystallization in semidilute solutions of ultra high molecular weight polyethylene

    DEFF Research Database (Denmark)

    Wingstrand, Sara Lindeblad; Imperiali, Luna; Stepanyan, Roman

    2018-01-01

    Abstract We investigate the influence of controlled uniaxial extension on various flow induced phenomena in semidilute solutions of ultra high molecular weight polyethylene (UHMwPE). Concentrations range from 9 w% to 29 w% and the choice of solvent is paraffin oil (PO). The start-up extensional b...

  9. Nanometer size wear debris generated from ultra high molecular weight polyethylene in vivo

    Czech Academy of Sciences Publication Activity Database

    Lapčíková, Monika; Šlouf, Miroslav; Dybal, Jiří; Zolotarevova, E.; Entlicher, G.; Pokorný, D.; Gallo, J.; Sosna, A.

    2009-01-01

    Roč. 266, 1-2 (2009), s. 349-355 ISSN 0043-1648 R&D Projects: GA MŠk 2B06096 Institutional research plan: CEZ:AV0Z40500505 Keywords : ultra high molecular weight polyethylene * nanometer size wear debris * morphology of wear particles Subject RIV: CD - Macromolecular Chemistry Impact factor: 1.771, year: 2009

  10. A 7-d exercise program increases high-molecular weight adiponectin in obese adults

    DEFF Research Database (Denmark)

    Kelly, Karen R; Blaszczak, Alecia; Haus, Jacob M

    2012-01-01

    High-molecular weight (HMW) adiponectin is the biologically active form of adiponectin and is related to enhanced insulin sensitivity and metabolic function. Previously, we found that 7 d of exercise improves insulin sensitivity in obese subjects; however, whether short-term exercise training...

  11. Dietary factors associated with plasma high molecular weight and total adiponectin levels in apparently healthy women

    NARCIS (Netherlands)

    Yannakoulia, Mary; Yiannakouris, Nikos; Melistas, Labros; Fappa, Evaggelia; Vidra, Nikoletta; Kontogianni, Meropi D; Mantzoros, Christos S

    2008-01-01

    OBJECTIVE: Our aim was to investigate associations between dietary factors and high molecular weight (HMW) as well as total adiponectin in a sample of apparently healthy adult Mediterranean women. DESIGN AND METHODS: Two hundred and twenty women were enrolled in this study. Anthropometric and body

  12. Chemical modification of high molecular weight polyethylene through gamma radiation for biomaterials applications

    International Nuclear Information System (INIS)

    Raposo, Matheus P.; Rocha, Marisa C.G.

    2015-01-01

    Ultra high molecular weight polyethylene has been used in the medical field due to its high mechanical properties compared to the other polymers. Its main application is in the development of orthopedic implants, which requires high resistance to abrasion. One of the most used methods is the introduction of crosslinks in the polymer through gamma irradiation. In order to prevent oxidation reactions, studies have been developed using tacoferol (vitamin E) as an antioxidant for the material. The ascorbic acid (vitamin C), however, has been appointed as a viable alternative for vitamin E. In this work, a high molecular weight polyethylene grade (HMWPE) and polyethylene samples formulated with vitamin C were submitted to gamma radiation. Thermodynamic-mechanical methods and gel content determinations were used to characterize the samples obtained. The sample containing 1% of vitamin C and irradiated with 50 KGy of gamma radiation presented the highest content of crosslinks. (author)

  13. High molecular weight DNA assembly in vivo for synthetic biology applications.

    Science.gov (United States)

    Juhas, Mario; Ajioka, James W

    2017-05-01

    DNA assembly is the key technology of the emerging interdisciplinary field of synthetic biology. While the assembly of smaller DNA fragments is usually performed in vitro, high molecular weight DNA molecules are assembled in vivo via homologous recombination in the host cell. Escherichia coli, Bacillus subtilis and Saccharomyces cerevisiae are the main hosts used for DNA assembly in vivo. Progress in DNA assembly over the last few years has paved the way for the construction of whole genomes. This review provides an update on recent synthetic biology advances with particular emphasis on high molecular weight DNA assembly in vivo in E. coli, B. subtilis and S. cerevisiae. Special attention is paid to the assembly of whole genomes, such as those of the first synthetic cell, synthetic yeast and minimal genomes.

  14. Use of electroporation for high-molecular-weight DNA-mediated gene transfer.

    Science.gov (United States)

    Jastreboff, M M; Ito, E; Bertino, J R; Narayanan, R

    1987-08-01

    Electroporation was used to introduce high-molecular-weight DNA into murine hematopoietic cells and NIH3T3 cells. CCRF-CEM cells were stably transfected with SV2NEO plasmid and the genomic DNA from G-418-resistant clones (greater than 65 kb) was introduced into mouse bone marrow and NIH3T3 cells by electroporation. NEO sequences and expression were detected in the hematopoietic tissues of lethally irradiated mice, with 24% of individual spleen colonies expressing NEO. The frequency of genomic DNA transfer into NIH3T3 cells was 0.25 X 10(-3). Electroporation thus offers a powerful mode of gene transfer not only of cloned genes but also of high-molecular-weight DNA into cells.

  15. Application of radiation grafting techniques to prepare the high molecular weight water-soluble polymer

    International Nuclear Information System (INIS)

    Le Hai; Nguyen Quoc Hien; Nguyen Tan Man; Truong Thi Hanh; Le Huu Tu; Tran Thi Tam; Pham Thi Sam; Pham Anh Tuan; Le Dinh Lang

    2003-01-01

    The results of the study on the preparation of the high molecular weight water-soluble polymers by radiation grafting and their properties is presented as follows: 1/ by radiation grafting, the molecular weight of PVA was increased 20 times and PAM was increased only 3 times; 2/ the thermal and medium stability of poly(vinyl alcohol) grafted with acrylamide was obviously improved. (LH)

  16. Evaluation of environmental degradation effects in morphology of ultra-high molecular weight polyethylene (UHMWPE) fibers

    International Nuclear Information System (INIS)

    Vivas, Viviane; Zylberberg, Marcel P.; Cardoso, Andre Luis V.; Pereira, Iaci M.; Weber, Ricardo P.; Suarez, Joao C. Miguez

    2015-01-01

    This study aims to evaluate changes in the morphology of ultra-high molecular weight polyethylene fiber (UHMWPE), before and after exposure to environmental agents. Fibers produced by two different manufacturers were analyzed. To characterize the morphology, we used the technique of small angle x-ray scattering (SAXS). The results demonstrate that the original morphology of the fibers was UHMWPE affected by the defects caused by exposure to environmental agents. (author)

  17. Urinary high molecular weight matrix metalloproteinases as non-invasive biomarker for detection of bladder cancer

    OpenAIRE

    Mohammed, Mohammed A; Seleim, Manar F; Abdalla, Mohga S; Sharada, Hayat M; Abdel Wahab, Abdel Hady A

    2013-01-01

    Background Matrix Metalloproteinases (MMPs) are key molecules for tumor growth, invasion and metastasis. Over-expression of different MMPs in tumor tissues can disturb the homeostasis and increase the level of various body fluids. Many MMPs including high molecular weights (HMWs) were detected in the urine of prostate and bladder cancer patients. Our aim here is to assess the usefulness of HMW MMPs as non invasive biomarkers in bilharzial bladder cancer in Egyptian patients. Methods The activ...

  18. Silver Nanoparticles Modification of Ultra High Molecular Weight Polyethylene in Non-Aqueous Medium

    OpenAIRE

    V. N. Glushko; L. I. Blokhina; E. E. Anisimova; M. V. Bogdanovskaya; V. I. Kozhukhov; T. A. Cherdyntseva

    2016-01-01

    A series of experiments for obtaining modified with silver nanoparticles ultra-high molecular weight polyethylene (UHMWPE) is done. Optimal precursors are silver trifluoroacetate, silver nitrate and silver methanesulfonate. Three variants of UHMWPE modification is studied: 1) the polyol synthesis, 2) polymer processing silver nanoparticle colloid and 3) reduction of silver salt solution in the UHMWPE polymer matrix. It is found that the last method is optimal. The specific surface of obtained...

  19. SDS-PAGE For Glutenins Protein of Some Durum Wheat Cultivars coming from Different Agro-Ecological Areas

    Directory of Open Access Journals (Sweden)

    Ahmed Salih Khalaf

    Full Text Available Ten durum wheat cultivars of different agro-ecological resources were involved, and subjected to electrophoresis for their glutenin subunits. Glutenin subunits resolution and coding based on relative mobility. Cultivars shown differences in migration patterns and categorized into different groups in accordance to possessing a particular subunits. Caronia and Haurani were similar as they possess subunits 6 + 15, while ACSAD 65, Simeto, and Waha possess subunits 7 + 8; but Korifla and Gidara 2 and Creso possess subunits 6 + 8 with an additional subunit 20 for the Creso. Lastly, Cham 5 and Om rabi 5 possess subunits 20.

  20. Radiation polymerization of acrylamide with super-high molecular weight in inverse emulsion

    International Nuclear Information System (INIS)

    Ye Qiang; Ge Xuewu; Xu Xiangling; Zhang Zhicheng

    1998-01-01

    The inverse emulsion polymerization of acrylamide has been studied with γ-ray initiation. Polyacrylamide with super high molecular weight over ten million (11 x 10 6 ), which is very important in application as flocculant, is obtained. In this work, some methods are taken to enhance the molecular weight as follows: (1) In order to prepare soluble polyacrylamide with super high molecular weight, the better conditions are: the emulsifier content is about 2% and the monomer concentration is about 20%∼24% in the composition of monomer emulsion, and the absorbed dose is about 500∼600 Gy. (2) Initiating with high dose rate and polymerizing with low dose rate can not only enhance the molecular weight of product, but also curtail the polymerizing time. (3) Stopping radiation when the conversion gets to about 10% and post-polymerizing outside the radiation source until the conversion gets to 82% can obtain polyacrylamide with super high molecular weight, and shorten the irradiation time as well

  1. High molecular weight poly(L-lactide) and poly(ethylene oxide) blends : Thermal characterization and physical properties

    NARCIS (Netherlands)

    Nijenhuis, AJ; Colstee, E; Grijpma, DW; Pennings, AJ

    1996-01-01

    The miscibility of high molecular weight poly(L-lactide) (PLLA) with high molecular weight poly(ethylene oxide) (PEG) was studied by differential scanning calorimetry. Ail blends containing up to 50 weight% PEO showed single glass transition temperatures. The PLLA and PEO melting temperatures were

  2. High molecular weight hyaluronan mediates the cancer resistance of the naked mole-rat

    Science.gov (United States)

    Tian, Xiao; Azpurua, Jorge; Hine, Christopher; Vaidya, Amita; Myakishev-Rempel, Max; Ablaeva, Julia; Mao, Zhiyong; Nevo, Eviatar; Gorbunova, Vera; Seluanov, Andrei

    2013-01-01

    The naked mole-rat displays exceptional longevity, with a maximum lifespan exceeding 30 years1–3. This is the longest reported lifespan for a rodent species and is especially striking considering the small body mass of the naked mole-rat. In comparison, a similarly sized house mouse has a maximum lifespan of 4 years4,5. In addition to their longevity, naked mole-rats show an unusual resistance to cancer. Multi-year observations of large naked mole-rat colonies did not detect a single incidence of cancer2,6. Here we identify a mechanism responsible for the naked mole-rat’s cancer resistance. We found that naked mole-rat fibroblasts secrete extremely high molecular weight hyaluronan (HA), which is over five times larger than human or mouse HA. This high molecular weight HA accumulates abundantly in naked mole rat tissues due to the decreased activity of HA-degrading enzymes and a unique sequence of hyaluronan synthase 2 (HAS2). Furthermore, the naked mole-rat cells are more sensitive to HA signaling, as the naked mole rat cells have a higher affinity to HA than the mouse or human cells. Perturbation of the signaling pathways sufficient for malignant transformation of mouse fibroblasts fails to transform naked mole-rat cells. However, once high molecular weight HA is removed by either knocking down HAS2 or overexpressing the HA-degrading enzyme, Hyal2, naked mole-rat cells become susceptible to malignant transformation and readily form tumors in mice. We speculate that naked mole-rats have evolved a higher concentration of HA in the skin to provide skin elasticity needed for life in underground tunnels. This trait may have then been co-opted to provide cancer resistance and longevity to this species. PMID:23783513

  3. Equal channel angular extrusion of ultra-high molecular weight polyethylene

    Energy Technology Data Exchange (ETDEWEB)

    Reinitz, Steven D., E-mail: Steven.D.Reinitz.TH@Dartmouth.edu; Engler, Alexander J.; Carlson, Evan M.; Van Citters, Douglas W.

    2016-10-01

    Ultra-high molecular weight polyethylene (UHMWPE), a common bearing surface in total joint arthroplasty, is subject to material property tradeoffs associated with conventional processing techniques. For orthopaedic applications, radiation-induced cross-linking is used to enhance the wear resistance of the material, but cross-linking also restricts relative chain movement in the amorphous regions and hence decreases toughness. Equal Channel Angular Extrusion (ECAE) is proposed as a novel mechanism by which entanglements can be introduced to the polymer bulk during consolidation, with the aim of imparting the same tribological benefits of conventional processing without complete inhibition of chain motion. ECAE processing at temperatures near the crystalline melt for UHMWPE produces (1) increased entanglements compared to control materials; (2) increasing entanglements with increasing temperature; and (3) mechanical properties between values for untreated polyethylene and for cross-linked polyethylene. These results support additional research in ECAE-processed UHMWPE for joint arthroplasty applications. - Highlights: • A new processing method for ultra-high molecular weight polyethylene is introduced. • The process produces a highly entangled polyethylene material. • Entanglements are hypothesized to enhance the wear resistance of polyethylene. • This process eliminates the trade-off between mechanical and wear properties.

  4. Single intra-articular injection of high molecular weight hyaluronic acid for hip osteoarthritis.

    Science.gov (United States)

    Rivera, Fabrizio

    2016-03-01

    Intra-articular (IA) injection of hyaluronic acid (HA) into the hip joint appears to be safe and well tolerated but only a small number of randomized clinical trials in humans has been published. The objective of this prospective study was to evaluate the efficacy and safety of a single IA injection of high-molecular-weight (2800 kDa) HA (Coxarthrum) for hip osteoarthritis. All patients received a single IA administration of 2.5 % sodium hyaluronate (75 mg/3 mL) of high molecular weight. Fluoroscopy requires an iodized contrast medium (iopamidol, 1 ml) which highlights the capsule before administering HA. Patients were evaluated before IA injection (T0), after 3 months, after 6 months and after 1 year from injection. Results were evaluated by the Brief Pain Inventory (BPI II), Harris Hip Score and a visual analog scale of pain (pain VAS). All treated patients were considered for statistical analysis. Two hundred seven patients were included at T0. The mean age was 67 years (range 46-81). Regarding BPI severity score, changes in pain between T0 and the three following visits were statistically highly significant (p injection of Coxarthrum is effective from the third month and that the results are stable or continue to improve up to 1 year. IV.

  5. Flexible mechanoprosthesis made from woven ultra-high-molecular-weight polyethylene fibres : proof of concept in a chronic sheep model

    NARCIS (Netherlands)

    Basir, Amir; Grobben, Remco B.; Cramer, Maarten Jan; van Herwaarden, Joost A.; Vink, Aryan; Pasterkamp, Gerard; Kluin, Jolanda; Gründeman, Paul F.

    2017-01-01

    OBJECTIVES: Ultra-high-molecular-weight polyethylene (UHMWPE) fibres are flexible, have high tensile strength, and platelet and bacterial adhesion is low. Therefore, UHMWPE may overcome limitations of current mechanical valves and bioprostheses. In this study, the biocompatibility and functionality

  6. Gliadin and glutenin polymorphism in durum wheat landraces and breeding varieties of Azerbaijan

    Directory of Open Access Journals (Sweden)

    Sadigov-Baykishi Hamlet

    2015-01-01

    Full Text Available Durum wheat genotypes including 7 landraces and 17 breeding varieties were studied. Polyacrylamide gel electrophoresis under acidic conditions of pH 3.1 was used to study gliadin and glutenin polymorphisms. In total, 32 gliadin and 8 high molecular weight glutenin alleles were identified. The contribution of B genome (58.5% to the allelic variation of durum wheat varieties was higher than of A genome. The cluster analysis delineated genotypes into four main clusters. According to cluster analysis, legitimacy identifying the distribution of botanical varieties through the tree was observed. The study confirms the suitability of biochemical markers for cultivar identification and genetic relation study in durum wheat genotypes.

  7. Ballistic behavior of ultra-high molecular weight polyethylene composite: effect of gamma radiation

    International Nuclear Information System (INIS)

    Alves, Andreia L. dos Santos; Nascimento, Lucio F.C.; Suarez, Joao C. Miguez; lucio2002bol.com.br

    2003-01-01

    Since World War II, textile composites have been used as ballistic armor. Ultra-high molecular weight polyethylene (UHMWPE) fibers are used in the production of armor materials. As they have been developed and commercialized only recently, there is not enough information about the effect of environmental agents in the ballistic performance of UHMWPE composites. In the present work, was evaluated the ballistic behavior of composite plates manufactured with UHMWPE fibers after exposure to gamma radiation. The ballistic tests results were related to the macromolecular alterations induced by the radiation through mechanical (hardness, impact and flexure) and physicochemical (Ftir/Mir. DSC and TGA) testing. It was observed that irradiation induces changes in the UHMWPE, degrading the ballistic performance of the composite. These results are presented and discussed. (author)

  8. Detection of high molecular weight proteins by MALDI imaging mass spectrometry.

    Science.gov (United States)

    Mainini, Veronica; Bovo, Giorgio; Chinello, Clizia; Gianazza, Erica; Grasso, Marco; Cattoretti, Giorgio; Magni, Fulvio

    2013-06-01

    MALDI imaging mass spectrometry (IMS) is a unique technology to explore the spatial distribution of biomolecules directly on tissues. It allows the in situ investigation of a large number of small proteins and peptides. Detection of high molecular weight proteins through MALDI IMS still represents an important challenge, as it would allow the direct investigation of the distribution of more proteins involved in biological processes, such as cytokines, enzymes, neuropeptide precursors and receptors. In this work we compare the traditional method performed with sinapinic acid with a comparable protocol using ferulic acid as the matrix. Data show a remarkable increase of signal acquisition in the mass range of 20k to 150k Th. Moreover, we report molecular images of biomolecules above 70k Th, demonstrating the possibility of expanding the application of this technology both in clinical investigations and basic science.

  9. Structural modifications of ultra-high molecular weight polyethylene (UHMWPE) processed in attritor type mill

    International Nuclear Information System (INIS)

    Gabriel, Melina C.; Carvalho, Benjamim de M.; Pinheiro, Luis A.; Cintho, Osvaldo M.; Capocchi, Jose D.T.; Kubaski, Evaldo T.

    2009-01-01

    Ultra-high molecular weight polyethylene (UHMWPE) is a polyethylene that has a high melt viscosity, hence its processing becomes very difficult. High-energy mechanical milling provides physical and chemical changes in polymers that have been studied recently. In order to study these changes in UHMWPE, powder of this polymer was mechanical milled in attritor type mill with a ball-to-powder weight ratio of 40:1 for 8 hours, varying the rotation speed: 200, 300, 400, 500 e 600 rpm. The polymer was characterized by scanning electron microscopy (SEM) and xray diffraction (XRD). From the XRD results it was noted that as the rotation speed increased the monoclinic phase also increased up to 500 rpm. For 600 rpm, the amount of monoclinic phase apparently decreased. At this rotation speed, the deformation rate probably increased the process temperature, allowing the monoclinic phase to return to its initial structural orthorhombic form. (author)

  10. Effects of inhaled high-molecular weight hyaluronan in inflammatory airway disease.

    Science.gov (United States)

    Lamas, Adelaida; Marshburn, Jamie; Stober, Vandy P; Donaldson, Scott H; Garantziotis, Stavros

    2016-10-03

    Cystic fibrosis (CF) is a chronic inflammatory disease that is affecting thousands of patients worldwide. Adjuvant anti-inflammatory treatment is an important component of cystic fibrosis treatment, and has shown promise in preserving lung function and prolonging life expectancy. Inhaled high molecular weight hyaluronan (HMW-HA) is reported to improve tolerability of hypertonic saline and thus increase compliance, and has been approved in some European countries for use as an adjunct to hypertonic saline treatment in cystic fibrosis. However, there are theoretical concerns that HMW-HA breakdown products may be pro-inflammatory. In this clinical pilot study we show that sputum cytokines in CF patients receiving HMW-HA are not increased, and therefore HMW-HA does not appear to adversely affect inflammatory status in CF airways.

  11. Possible hypoglycemic effect of Aloe vera L. high molecular weight fractions on type 2 diabetic patients

    Science.gov (United States)

    Yagi, Akira; Hegazy, Sahar; Kabbash, Amal; Wahab, Engy Abd-El

    2009-01-01

    Aloe vera L. high molecular weight fractions (AHM) containing less than 10 ppm of barbaloin and polysaccharide (MW: 1000 kDa) with glycoprotein, verectin (MW: 29 kDa), were prepared by patented hyper-dry system in combination of freeze–dry technique with microwave and far infrared radiation. AHM produced significant decrease in blood glucose level sustained for 6 weeks of the start of the study. Significant decrease in triglycerides was only observed 4 weeks after treatment and continued thereafter. No deterious effects on kidney and liver functions were apparent. Treatment of diabetic patients with AHM may relief vascular complications probably via activation of immunosystem. PMID:23964163

  12. Elevated levels of high-molecular-weight adiponectin in type 1 diabetes

    DEFF Research Database (Denmark)

    Leth, H.; Andersen, K.K.; Frystyk, J.

    2008-01-01

    BACKGROUND: Several studies have shown that type 1 diabetic patients have elevated total levels of the adipocyte-derived adipocytokine adiponectin. However, adiponectin circulates in three different subforms, and the high-molecular-weight (HMW) subform is believed to be the primary biologically...... active form. The effects of the medium-molecular-weight (MMW) subform and the low-molecular-weight (LMW) subform are still unresolved. PURPOSE: The objective of the study was to investigate the distribution of the three molecular subforms of adiponectin in well-characterized groups of type 1 diabetics...... with varying degrees of nephropathy as well as in healthy control subjects. STUDY POPULATION: Two hundred seven individuals were included: 58 type 1 diabetics with normoalbuminuria, 46 with microalbuminuria, 46 with macroalbuminuria, and 57 matched controls. METHODS: The HMW, MMW, and LMW subforms were...

  13. Sequence and properties of HMW subunit 1Bx20 from pasta wheat (Triticum durum) which is associated with poor end use properties.

    Science.gov (United States)

    Shewry, P R; Gilbert, S M; Savage, A W J; Tatham, A S; Wan, Y-F; Belton, P S; Wellner, N; D'Ovidio, R; Békés, F; Halford, N G

    2003-02-01

    The gene encoding high-molecular-weight (HMW) subunit 1Bx20 was isolated from durum wheat cv. Lira. It encodes a mature protein of 774 amino acid residues with an M(r) of 83,913. Comparison with the sequence of subunit 1Bx7 showed over 96% identity, the main difference being the substitution of two cysteine residues in the N-terminal domain of subunit 1Bx7 with tyrosine residues in 1Bx20. Comparison of the structures and stabilities of the two subunits purified from wheat using Fourier-transform infra-red and circular dichroism spectroscopy showed no significant differences. However, incorporation of subunit 1Bx7 into a base flour gave increased dough strength and stability measured by Mixograph analysis, while incorporation of subunit 1Bx20 resulted in small positive or negative effects on the parameters measured. It is concluded that the different effects of the two subunits could relate to the differences in their cysteine contents, thereby affecting the cross-linking and hence properties of the glutenin polymers.

  14. Effect of high and low molecular weight glutenin subunits, and subunits of gliadin on physicochemical parameters of different wheat genotypes Efeito das subunidades de glutenina de alto e baixo peso molecular e das subunidades de gliadina sobre os parâmetros físico-químicos de diferentes genótipos de trigo

    Directory of Open Access Journals (Sweden)

    Mariana Souza Costa

    2013-02-01

    Full Text Available Identification of functional properties of wheat flour by specific tests allows genotypes with appropriate characteristics to be selected for specific industrial uses. The objective of wheat breeding programs is to improve the quality of germplasm bank in order to be able to develop wheat with suitable gluten strength and extensibility for bread making. The aim of this study was to evaluate 16 wheat genotypes by correlating both glutenin subunits of high and low molecular weight and gliadin subunits with the physicochemical characteristics of the grain. Protein content, sedimentation volume, sedimentation index, and falling number values were analyzed after the grains were milled. Hectoliter weight and mass of 1000 seeds were also determined. The glutenin and gliadin subunits were separated using polyacrylamide gel in the presence of sodium dodecyl sulfate. The data were evaluated using variance analysis, Pearson's correlation, principal component analysis, and cluster analysis. The IPR 85, IPR Catuara TM, T 091015, and T 091069 genotypes stood out from the others, which indicate their possibly superior grain quality with higher sedimentation volume, higher sedimentation index, and higher mass of 1000 seeds; these genotypes possessed the subunits 1 (Glu-A1, 5 + 10 (Glu-D1, c (Glu-A3, and b (Glu-B3, with exception of T 091069 genotype that possessed the g allele instead of b in the Glu-B3.A identificação das propriedades funcionais de farinhas de trigo através de testes específicos possibilita selecionar genótipos de trigo com características adequadas a cada uso industrial. O principal objetivo dos programas de melhoramento genético é melhorar a qualidade do banco de germoplasma para que seja possível desenvolver trigos com força de glúten e extensibilidade adequadas para produção de produtos panificáveis. O objetivo deste estudo foi avaliar 16 genótipos de trigo correlacionando as subunidades de gluteninas de alto e baixo peso

  15. Photoluminescence of ultra-high molecular weight polyethylene modified by fast atom bombardment

    International Nuclear Information System (INIS)

    Toth, S.; Fuele, M.; Veres, M.; Pocsik, I.; Koos, M.; Toth, A.; Ujvari, T.; Bertoti, I.

    2006-01-01

    An increase in the application potential of the ultra-high molecular weight polyethylene (UHMWPE) may be achieved by producing a hard, wear resistant carbonaceous modified surface layer on it. In this study the surface of UHMWPE samples was treated by 1 keV N, H and He fast atom bombardment (FAB) to obtain amorphous carbon surface layer which produces an enhancement of microhardness. The untreated and FAB-modified samples were investigated by photoluminescence, infrared, Raman and optical absorption spectroscopy. The FAB-treatment caused a nearly complete disappearance of the characteristic luminescence bands of UHMWPE (at 335, 351, 363 and 381 nm), the appearance of new bands at 459 and 495 nm due to the formation of new recombination levels in the FAB-treated samples. The remarkable decrease in integrated luminescence intensity indicates the appearance of new non-radiative recombination levels caused by FAB treatment. Structural modifications in FAB treated samples result in the development of structural arrangement containing sp 2 bonded carbon sites in rings or chains of different sizes and the electronic levels corresponding to these structural elements are situated in the forbidden gap in the electronic density of states which brings forth the observed changes of the photoluminescence properties

  16. Macroradical reaction in ultra-high molecular weight polyethylene in the presence of vitamin E

    International Nuclear Information System (INIS)

    Jahan, M.S.; Walters, B.M.

    2011-01-01

    Free radical measurements in compression molded ultra-high molecular weight polyethylene (UHMWPE), which contained vitamin E (α-tocopherol (α-T)), was performed using electron spin resonance (ESR) technique in air at room temperature following gamma irradiation (25-32 kGy) in N 2 . The vitamin E was incorporated into one set of samples by blending UHMWPE resin with vitamin E (1 and 10 wt%), then compression molded into a solid and then irradiated. Another set of samples had vitamin E incorporated into them by diffusing vitamin E at 100 o C for 2 h after irradiation. Compared to a control (with no vitamin E), the vitamin E-containing UHMWPE (α-TPE) samples suffered a partial loss of PE radicals, but this loss only occurred during or immediately after irradiation (before exposure to air). Subsequently, when all blended samples were exposed to air, the remaining radicals in each sample decayed to the well-known OIR, R1 (- · CH-[CH=CH-] m -) and R2 ( · OCH-[CH=CH-] m -) radicals. However, because of the initial loss or partial quenching, α-TPE produced a lower concentration of OIR (measured over a four-year period), but no difference was found between 1% and 10% α-TPEs. In the diffused α-TPE, similar OIR was also found when tested after four months of post-treatment exposure to air.

  17. Macroradical reaction in ultra-high molecular weight polyethylene in the presence of vitamin E

    Energy Technology Data Exchange (ETDEWEB)

    Jahan, M.S., E-mail: mjahan@memphis.ed [Department of Physics, Biomaterials Research Laboratory, University of Memphis, 216 Manning Hall, Memphis, TN 38152 (United States); Walters, B.M. [Department of Physics, Biomaterials Research Laboratory, University of Memphis, 216 Manning Hall, Memphis, TN 38152 (United States)

    2011-02-15

    Free radical measurements in compression molded ultra-high molecular weight polyethylene (UHMWPE), which contained vitamin E ({alpha}-tocopherol ({alpha}-T)), was performed using electron spin resonance (ESR) technique in air at room temperature following gamma irradiation (25-32 kGy) in N{sub 2}. The vitamin E was incorporated into one set of samples by blending UHMWPE resin with vitamin E (1 and 10 wt%), then compression molded into a solid and then irradiated. Another set of samples had vitamin E incorporated into them by diffusing vitamin E at 100 {sup o}C for 2 h after irradiation. Compared to a control (with no vitamin E), the vitamin E-containing UHMWPE ({alpha}-TPE) samples suffered a partial loss of PE radicals, but this loss only occurred during or immediately after irradiation (before exposure to air). Subsequently, when all blended samples were exposed to air, the remaining radicals in each sample decayed to the well-known OIR, R1 (-{sup {center_dot}C}H-[CH=CH-]{sub m}-) and R2 ({sup {center_dot}O}CH-[CH=CH-]{sub m}-) radicals. However, because of the initial loss or partial quenching, {alpha}-TPE produced a lower concentration of OIR (measured over a four-year period), but no difference was found between 1% and 10% {alpha}-TPEs. In the diffused {alpha}-TPE, similar OIR was also found when tested after four months of post-treatment exposure to air.

  18. Synthesis of polymer gel electrolyte with high molecular weight poly(methyl methacrylate)-clay nanocomposite

    International Nuclear Information System (INIS)

    Meneghetti, Paulo; Qutubuddin, Syed; Webber, Andrew

    2004-01-01

    Polymer nanocomposite gel electrolytes consisting of high molecular weight poly(methyl methacrylate) PMMA-clay nanocomposite, ethylene carbonate (EC)/propylene carbonate (PC) as plasticizer, and LiClO 4 electrolyte are reported. Montmorillonite clay was ion exchanged with a zwitterionic surfactant (octadecyl dimethyl betaine) and dispersed in methyl methacrylate, which was then polymerized to synthesize PMMA-clay nanocomposites. The nanocomposite was dissolved in a mixture of EC/PC with LiClO 4 , heated and pressed to obtain polymer gel electrolyte. X-ray diffraction (XRD) of the gels indicated intercalated clay structure with d-spacings of 2.85 and 1.40 nm. In the gel containing plasticizer, the clay galleries shrink suggesting intercalation rather than partial exfoliation observed in the PMMA-clay nanocomposite. Ionic conductivity varied slightly and exhibited a maximum value of 8 x 10 -4 S/cm at clay content of 1.5 wt.%. The activation energy was determined by modeling the conductivity with a Vogel-Tamman-Fulcher expression. The clay layers are primarily trapped inside the polymer matrix. Consequently, the polymer does not interact significantly with LiClO 4 electrolyte as shown by FTIR. The presence of the clay increased the glass transition temperature (Tg) of the gel as determined by differential scanning calorimetry. The PMMA nanocomposite gel electrolyte shows a stable lithium interfacial resistance over time, which is a key factor for use in electrochemical applications

  19. Laser surface modification of ultra-high-molecular-weight polyethylene (UHMWPE) for biomedical applications

    International Nuclear Information System (INIS)

    Riveiro, A.; Soto, R.; Val, J. del; Comesaña, R.; Boutinguiza, M.; Quintero, F.; Lusquiños, F.; Pou, J.

    2014-01-01

    Ultra-high-molecular-weight polyethylene (UHMWPE) is a synthetic polymer used for biomedical applications because of its high impact resistance, ductility and stability in contact with physiological fluids. Therefore, this material is being used in human orthopedic implants such as total hip or knee replacements. Surface modification of this material relates to changes on its chemistry, microstructure, roughness, and topography, all influencing its biological response. Surface treatment of UHMWPE is very difficult due to its high melt viscosity. This work presents a systematic approach to discern the role of different laser wavelengths (λ = 1064, 532, and 355 nm) on the surface modification of carbon coated UHMWPE samples. Influence of laser processing conditions (irradiance, pulse frequency, scanning speed, and spot overlapping) on the surface properties of this material was determined using an advanced statistical planning of experiments. A full factorial design of experiments was used to find the main effects of the processing parameters. The obtained results indicate the way to maximize surface properties which largely influence cell–material interaction.

  20. Surface functionalization of solid state ultra-high molecular weight polyethylene through chemical grafting

    Science.gov (United States)

    Sherazi, Tauqir A.; Rehman, Tayyiba; Naqvi, Syed Ali Raza; Shaikh, Ahson Jabbar; Shahzad, Sohail Anjum; Abbas, Ghazanfar; Raza, Rizwan; Waseem, Amir

    2015-12-01

    The surface of ultra-high molecular weight polyethylene (UHMWPE) powder was functionalized with styrene using chemical grafting technique. The grafting process was initiated through radical generation on base polymer matrix in the solid state by sodium thiosulfate, while peroxides formed at radical sites during this process were dissociated by ceric ammonium nitrate. Various factors were optimized and reasonably high level of monomer grafting was achieved, i.e., 15.6%. The effect of different acids as additive and divinyl benzene (DVB) as a cross-linking agent was also studied. Post-grafting sulfonation was conducted to introduce the ionic moieties to the grafted polymer. Ion-exchange capacity (IEC) was measured experimentally and is found to be 1.04 meq g-1, which is in close agreement with the theoretical IEC values. The chemical structure of grafted and functionalized polymer was characterized by attenuated total reflection infrared spectroscopy (ATR-FTIR) and thermal properties were investigated by thermo gravimetric analysis (TGA) and differential scanning calorimetry (DSC). Thermal analysis depicts that the presence of radicals on the polymer chain accelerates the thermal decomposition process. The results signify that the chemical grafting is an effective tool for substantial surface modification and subsequent functionalization of polyethylene.

  1. Risk organ preservation technique in reirradiation. Injection of native type high molecular weight hyaluronate

    International Nuclear Information System (INIS)

    Kishi, Kazushi; Noda, Yasutaka; Tanaka, Kayo

    2012-01-01

    We developed a practical method of brachytherapy effective for reirradiation, using hyaluronate gel injection to separate the target to be intensively. We intended curative reirradiation with preserving organs at risk. Native-type high molecular weight hyaluronate that has an ideal biologic property to reduce inflammation via inhibitory activity to CD44 and other inflammation receptors was injected to create space during radiotherapy. Rectum, small intestines, skin, and various normal organs are effectively separated and thus eradicative re-irradiation was carried out in safe. We reviewed 55 patients who required the method. There were no complications related to the procedure. Prescribed dose to 100% of the planning target volume (PTV) was 77.93 Gy (range 54-92) equivalent at alpha/beta ratio of 3, with much higher subvolume effect, and the saving effects were enhanced at 3.35 times in the average of those without gel injection. In conclusion, the interventional brachytherapy with hyaluronate gel injection provided 3.35 times safer dose-escalated and eradicative treatment in reirradiation. (author)

  2. Purification of High Molecular Weight Genomic DNA from Powdery Mildew for Long-Read Sequencing.

    Science.gov (United States)

    Feehan, Joanna M; Scheibel, Katherine E; Bourras, Salim; Underwood, William; Keller, Beat; Somerville, Shauna C

    2017-03-31

    The powdery mildew fungi are a group of economically important fungal plant pathogens. Relatively little is known about the molecular biology and genetics of these pathogens, in part due to a lack of well-developed genetic and genomic resources. These organisms have large, repetitive genomes, which have made genome sequencing and assembly prohibitively difficult. Here, we describe methods for the collection, extraction, purification and quality control assessment of high molecular weight genomic DNA from one powdery mildew species, Golovinomyces cichoracearum. The protocol described includes mechanical disruption of spores followed by an optimized phenol/chloroform genomic DNA extraction. A typical yield was 7 µg DNA per 150 mg conidia. The genomic DNA that is isolated using this procedure is suitable for long-read sequencing (i.e., > 48.5 kbp). Quality control measures to ensure the size, yield, and purity of the genomic DNA are also described in this method. Sequencing of the genomic DNA of the quality described here will allow for the assembly and comparison of multiple powdery mildew genomes, which in turn will lead to a better understanding and improved control of this agricultural pathogen.

  3. Extraction of High Molecular Weight DNA from Fungal Rust Spores for Long Read Sequencing.

    Science.gov (United States)

    Schwessinger, Benjamin; Rathjen, John P

    2017-01-01

    Wheat rust fungi are complex organisms with a complete life cycle that involves two different host plants and five different spore types. During the asexual infection cycle on wheat, rusts produce massive amounts of dikaryotic urediniospores. These spores are dikaryotic (two nuclei) with each nucleus containing one haploid genome. This dikaryotic state is likely to contribute to their evolutionary success, making them some of the major wheat pathogens globally. Despite this, most published wheat rust genomes are highly fragmented and contain very little haplotype-specific sequence information. Current long-read sequencing technologies hold great promise to provide more contiguous and haplotype-phased genome assemblies. Long reads are able to span repetitive regions and phase structural differences between the haplomes. This increased genome resolution enables the identification of complex loci and the study of genome evolution beyond simple nucleotide polymorphisms. Long-read technologies require pure high molecular weight DNA as an input for sequencing. Here, we describe a DNA extraction protocol for rust spores that yields pure double-stranded DNA molecules with molecular weight of >50 kilo-base pairs (kbp). The isolated DNA is of sufficient purity for PacBio long-read sequencing, but may require additional purification for other sequencing technologies such as Nanopore and 10× Genomics.

  4. Laser surface modification of ultra-high-molecular-weight polyethylene (UHMWPE) for biomedical applications

    Energy Technology Data Exchange (ETDEWEB)

    Riveiro, A., E-mail: ariveiro@uvigo.es [Applied Physics Department, University of Vigo, ETSII, Lagoas-Marcosende, 9, Vigo 36310 (Spain); Centro Universitario de la Defensa, Escuela Naval Militar, Plaza de España 2, 36920 Marín (Spain); Soto, R.; Val, J. del; Comesaña, R.; Boutinguiza, M.; Quintero, F.; Lusquiños, F.; Pou, J. [Applied Physics Department, University of Vigo, ETSII, Lagoas-Marcosende, 9, Vigo 36310 (Spain)

    2014-05-01

    Ultra-high-molecular-weight polyethylene (UHMWPE) is a synthetic polymer used for biomedical applications because of its high impact resistance, ductility and stability in contact with physiological fluids. Therefore, this material is being used in human orthopedic implants such as total hip or knee replacements. Surface modification of this material relates to changes on its chemistry, microstructure, roughness, and topography, all influencing its biological response. Surface treatment of UHMWPE is very difficult due to its high melt viscosity. This work presents a systematic approach to discern the role of different laser wavelengths (λ = 1064, 532, and 355 nm) on the surface modification of carbon coated UHMWPE samples. Influence of laser processing conditions (irradiance, pulse frequency, scanning speed, and spot overlapping) on the surface properties of this material was determined using an advanced statistical planning of experiments. A full factorial design of experiments was used to find the main effects of the processing parameters. The obtained results indicate the way to maximize surface properties which largely influence cell–material interaction.

  5. Extrudable polymer-polymer composites based on ultra-high molecular weight polyethylene

    Science.gov (United States)

    Panin, S. V.; Kornienko, L. A.; Alexenko, V. O.; Buslovich, D. G.; Dontsov, Yu. V.

    2017-12-01

    Mechanical and tribotechnical characteristics of polymer-polymeric composites of UHMWPE are studied with the aim of developing extrudable, wear-resistant, self-lubricant polymer mixtures for Additive Manufacturing (AM). The motivation of the study is their further application as feedstocks for 3D printing. Blends of UHMWPE with graft- and block copolymers of low-density polyethylene (HDPE-g-VTMS, HDPE-g-SMA, HDPE-b-EVA), polypropylene (PP), block copolymers of polypropylene and polyamide with linear low density polyethylene (PP-b-LLDPE, PA-b-LLDPE), as well as cross-linked polyethylene (PEX-b), are examined. The choice of compatible polymer components for an ultra- high molecular weight matrix for increasing processability (extrudability) is motivated by the search for commercially available and efficient additives aimed at developing wear-resistant extrudable polymer composites for additive manufacturing. The extrudability, mechanical properties and wear resistance of UHMWPE-based polymer-polymeric composites under sliding friction with different velocities and loads are studied.

  6. Conductivity, dielectric behavior and FTIR studies of high molecular weight poly(vinylchloride)-lithium triflate polymer electrolytes

    International Nuclear Information System (INIS)

    Ramesh, S.; Chai, M.F.

    2007-01-01

    Thin films of high molecular weight polyvinyl chloride (PVC) with lithium triflate (LiCF 3 SO 3 ) salt were prepared by solution casting method. The ionic conductivity and dielectric measurements were carried out on these films over a wide frequency regime at various temperatures. The conductivity-temperature plots were found to obey classical Arrhenius relationship. The dielectric behavior was analysed using dielectric permittivity and dielectric modulus of the samples. FTIR studies show some simple overlapping and shift in peaks between high molecular weight polyvinyl chloride (PVC) with lithium triflate (LiCF 3 SO 3 ) salt in the polymer electrolyte complexes

  7. Conductivity, dielectric behavior and FTIR studies of high molecular weight poly(vinylchloride)-lithium triflate polymer electrolytes

    Energy Technology Data Exchange (ETDEWEB)

    Ramesh, S. [Faculty of Engineering and Science, Universiti Tunku Abdul Rahman, Setapak, 53300 Kuala Lumpur (Malaysia)]. E-mail: ramesh@mail.utar.edu.my; Chai, M.F. [Faculty of Engineering and Science, Universiti Tunku Abdul Rahman, Setapak, 53300 Kuala Lumpur (Malaysia)

    2007-05-15

    Thin films of high molecular weight polyvinyl chloride (PVC) with lithium triflate (LiCF{sub 3}SO{sub 3}) salt were prepared by solution casting method. The ionic conductivity and dielectric measurements were carried out on these films over a wide frequency regime at various temperatures. The conductivity-temperature plots were found to obey classical Arrhenius relationship. The dielectric behavior was analysed using dielectric permittivity and dielectric modulus of the samples. FTIR studies show some simple overlapping and shift in peaks between high molecular weight polyvinyl chloride (PVC) with lithium triflate (LiCF{sub 3}SO{sub 3}) salt in the polymer electrolyte complexes.

  8. Dosimetric features and kinetic analysis of thermoluminescence from ultra-high molecular weight polyethylene

    Science.gov (United States)

    Chithambo, M. L.

    2012-08-01

    Thermoluminescence (TL) from beta irradiated ultra-high molecular weight polyethylene has been studied for measurements between 30 and 200 °C. An aliquot studied in this work produced TL glow curves consisting of two peaks, the main peak at 88 °C and a weaker intensity peak at 148 °C for heating at 1 °C s-1 following an excitation dose of 215 Gy. The position of the main peak is poorly reproducible for heating rates of 0.2 and 0.6 °C s-1 investigated with the peak position decreasing when the sample is freshly irradiated and the TL re-measured. The said change in peak position is however less of an effect for measurements made at 1 °C s-1 with the peak position being fairly reproducible in this case. Further measurements of the dosimetric properties of ultra-high molecular weight polyethylene showed that its dose response is linear from 26 Gy to about 161 Gy but exhibits slower growth in intensity with dose from about 860 Gy after regions of sub- and supra-linearity in between. If the TL is not measured immediately after irradiation, the signal fades with the delay approximately exponentially. In addition, a number of tests including phosphorescence analysis showed the possibility that the order of kinetics might not be unique but sensitive to several factors including measurement temperature. Thus for instance, the dependence of the peak position on the stop temperature in the partial heating procedure Tm - Tstop implied first-order kinetics but analysis of the geometrical factor μg for the same set of data gave μg = 0.46 ± 0.03 a value corresponding to characteristics somewhat intermediate between first and second order. In comparison, the results of analysis of the phosphorescence recorded at several temperatures on the rising edge of the main peak were only in agreement for measurements at 40 °C with general-order analysis suggesting second-order kinetics apply as did TL-like transformation of the monotonic phosphorescence decay. Both results were

  9. Topochemical approach to efficiently produce main-chain poly(bile acid)s with high molecular weights.

    Science.gov (United States)

    Li, Weina; Li, Xuesong; Zhu, Wei; Li, Changxu; Xu, Dan; Ju, Yong; Li, Guangtao

    2011-07-21

    Based on a topochemical approach, a strategy for efficiently producing main-chain poly(bile acid)s in the solid state was developed. This strategy allows for facile and scalable synthesis of main-chain poly(bile acid)s not only with high molecular weights, but also with quantitative conversions and yields.

  10. Flexible mechanoprosthesis made from woven ultra-high-molecular-weight polyethylene fibres: proof of concept in a chronic sheep model

    NARCIS (Netherlands)

    Basir, Amir; Grobben, Remco B.; Cramer, Maarten Jan; van Herwaarden, Joost A.; Vink, Aryan; Pasterkamp, Gerard; Kluin, Jolanda; Gründeman, Paul F.

    2017-01-01

    OBJECTIVES: Ultra-high-molecular-weight polyethylene (UHMWPE) fibres are flexible, have high tensile strength, and platelet and bacterial adhesion is low. Therefore, UHMWPE may overcome limitations of current mechanical valves and bioprostheses. In this study, the bio-compatibility and functionality

  11. Mucin Agarose Gel Electrophoresis: Western Blotting for High-molecular-weight Glycoproteins.

    Science.gov (United States)

    Ramsey, Kathryn A; Rushton, Zachary L; Ehre, Camille

    2016-06-14

    Mucins, the heavily-glycosylated proteins lining mucosal surfaces, have evolved as a key component of innate defense by protecting the epithelium against invading pathogens. The main role of these macromolecules is to facilitate particle trapping and clearance while promoting lubrication of the mucosa. During protein synthesis, mucins undergo intense O-glycosylation and multimerization, which dramatically increase the mass and size of these molecules. These post-translational modifications are critical for the viscoelastic properties of mucus. As a result of the complex biochemical and biophysical nature of these molecules, working with mucins provides many challenges that cannot be overcome by conventional protein analysis methods. For instance, their high-molecular-weight prevents electrophoretic migration via regular polyacrylamide gels and their sticky nature causes adhesion to experimental tubing. However, investigating the role of mucins in health (e.g., maintaining mucosal integrity) and disease (e.g., hyperconcentration, mucostasis, cancer) has recently gained interest and mucins are being investigated as a therapeutic target. A better understanding of the production and function of mucin macromolecules may lead to novel pharmaceutical approaches, e.g., inhibitors of mucin granule exocytosis and/or mucolytic agents. Therefore, consistent and reliable protocols to investigate mucin biology are critical for scientific advancement. Here, we describe conventional methods to separate mucin macromolecules by electrophoresis using an agarose gel, transfer protein into nitrocellulose membrane, and detect signal with mucin-specific antibodies as well as infrared fluorescent gel reader. These techniques are widely applicable to determine mucin quantitation, multimerization and to test the effects of pharmacological compounds on mucins.

  12. Total and high molecular weight adiponectin are elevated in patients with Laron syndrome despite marked obesity.

    Science.gov (United States)

    Kanety, Hannah; Hemi, Rina; Ginsberg, Shira; Pariente, Clara; Yissachar, Eleanor; Barhod, Ehud; Funahashi, Tohru; Laron, Zvi

    2009-12-01

    Patients with Laron syndrome (LS; primary GH insensitivity) caused by molecular defects of the GH receptor gene, are characterized by dwarfism, profound obesity, and hyperlipidemia. The aim of the current study was to evaluate adiponectin levels in LS, as obesity is known to be associated with low adiponectin. We studied nine untreated LS adult patients (5 males, 4 females) and six girls with LS receiving once-daily treatment by IGF1. Total and high molecular weight (HMW) adiponectin levels, adiponectin multimers distribution, and metabolic indices were analyzed in serum samples obtained during several years of follow-up. Adiponectin levels in the severely obese adult LS patients (percent body fat; females 61.0+/-2.5%, males 40.6+/-8.1%) were two- to three-fold higher than those reported for subjects of corresponding age, gender and degree of adiposity. Total adiponectin was significantly higher in females compared with males (21.4+/-3.5 vs 10.2+/-4.6 microg/ml, P<0.001). The elevated adiponectin in LS subjects was associated with an increased abundance of the HMW isoform, and positively correlated with body fat percentage (r=0.65, P=0.017) and leptin (r=0.65, P=0.012). There was no correlation between adiponectin levels (total and HMW) and the degree of insulin resistance in LS subjects or their blood lipids levels. Adiponectin was also high in young girls with LS (22.9+/-7.4 microg/ml) and did not change during long-term IGF1 replacement therapy. Adiponectin hypersecretion in LS, despite profound obesity, suggests that GH activity may negatively impact adiponectin secretion from adipocytes.

  13. Equilibrating high-molecular-weight symmetric and miscible polymer blends with hierarchical back-mapping

    Science.gov (United States)

    Ohkuma, Takahiro; Kremer, Kurt; Daoulas, Kostas

    2018-05-01

    Understanding properties of polymer alloys with computer simulations frequently requires equilibration of samples comprised of microscopically described long molecules. We present the extension of an efficient hierarchical backmapping strategy, initially developed for homopolymer melts, to equilibrate high-molecular-weight binary blends. These mixtures present significant interest for practical applications and fundamental polymer physics. In our approach, the blend is coarse-grained into models representing polymers as chains of soft blobs. Each blob stands for a subchain with N b microscopic monomers. A hierarchy of blob-based models with different resolution is obtained by varying N b. First the model with the largest N b is used to obtain an equilibrated blend. This configuration is sequentially fine-grained, reinserting at each step the degrees of freedom of the next in the hierarchy blob-based model. Once the blob-based description is sufficiently detailed, the microscopic monomers are reinserted. The hard excluded volume is recovered through a push-off procedure and the sample is re-equilibrated with molecular dynamics (MD), requiring relaxation on the order of the entanglement time. For the initial method development we focus on miscible blends described on microscopic level through a generic bead-spring model, which reproduces hard excluded volume, strong covalent bonds, and realistic liquid density. The blended homopolymers are symmetric with respect to molecular architecture and liquid structure. To parameterize the blob-based models and validate equilibration of backmapped samples, we obtain reference data from independent hybrid simulations combining MD and identity exchange Monte Carlo moves, taking advantage of the symmetry of the blends. The potential of the backmapping strategy is demonstrated by equilibrating blend samples with different degree of miscibility, containing 500 chains with 1000 monomers each. Equilibration is verified by comparing

  14. Early weight changes after birth and serum high-molecular-weight adiponectin level in preterm infants.

    Science.gov (United States)

    Yoshida, Tomohide; Nagasaki, Hiraku; Asato, Yoshihide; Ohta, Takao

    2011-12-01

    Extra-uterine growth retardation (EUGR) is associated with an increased risk for cardiometabolic diseases later in life. The aim of the present study was to examine the relationship between early weight change after birth in preterm infants and adiponectin (adn) multimeric complexes. Subjects included 28 preterm infants born between weeks 24 and 33 of gestation. Serum adn multimeric complexes and the anthropometric parameters were measured in preterm infants at birth and at corrected term. Bodyweight (BW) decreased during the first week of life, with birthweight restored at approximately 19 days after birth. Nineteen of the subjects had EUGR at corrected term. Total (T)-adn, high-molecular-weight (H)-adn, and the ratio of H-adn to T-adn (H/T-adn) were significantly elevated at corrected term than at birth. Postmenstrual age, birthweight, birth length and lowest BW after birth were positively correlated with H-adn and H/T-adn. Weight reduction after birth was negatively correlated with H-adn. Age to restore birthweight was negatively correlated with T-adn, H-adn and H/T-adn. Stepwise multiple regression analysis indicated age to restore birthweight as the major predictor of T-adn and H-adn. Early weight changes after birth may alter serum adn level in preterm infants at corrected term. The appropriate nutritional support in the early postnatal period could reduce the prevalence of EUGR and the future risk for cardiometabolic diseases. © 2011 The Authors. Pediatrics International © 2011 Japan Pediatric Society.

  15. Characterization of high molecular weight cadmium species in contaminated vegetable food

    Energy Technology Data Exchange (ETDEWEB)

    Guenther, K.; Kastenholz, B. [Forschungszentrum Juelich GmbH (Germany). Inst. fuer Chemie und Dynamik der Geosphaere 7: Angewandte Physikalische Chemie; Ji, G. [Bonn Univ. (Germany). Lehrstuhl fuer Lebensmittelwissenschaft und Lebensmittelchemie

    2000-10-01

    Spinach and radish grown from seeds were each contaminated with 4 different amounts of cadmium. After a cell breakdown of the eatable parts and centrifugation of the resulting homogenates all supernatants (cytosols) were separated by gel permeation chromatography (GPC). The size-range of the GPC method used was about 20-8000 kDa for globular proteins. The high molecular weight (HMW-Cd-SP, 150-700 kDa) and the low molecular weight Cd species (LMW-Cd-SP, < 150 kDa) in all plant cytosols eluted at about the same retention volume by GPC. The most important Cd binding form in the cytosols of all plants was found to be HMW-Cd-SP. The Cd elution maxima were detected in the range of about 200 kDa. The Cd determinations were performed with ET-AAS by means of matrix modifier. By incubating chosen cytosols with a proteinase before the GPC it was verified that the HMW-Cd-SP in both vegetables are Cd proteins. The molar proportions protein/Cd were about 2-6 in the respective GPC fractions of the HMW-Cd-SP of the highest contaminated plants. The GPC fractions of the HMW-Cd-SP of spinach and radish were further separated by a preparative, native and continuous polyacrylamide gel electrophoresis (PAGE) method. At pH 8 the species were negatively charged, had only a small UV-absorption at 280 nm and showed a very similar elution behavior in all analyzed cytosols. Therefore, we suppose that the HMW-Cd-SP of these two different vegetable foodstuffs have a very similar chemical structure. (orig.)

  16. High molecular weight of polysaccharides from Hericium erinaceus against amyloid beta-induced neurotoxicity.

    Science.gov (United States)

    Cheng, Jai-Hong; Tsai, Chia-Ling; Lien, Yi-Yang; Lee, Meng-Shiou; Sheu, Shyang-Chwen

    2016-06-07

    Hericium erinaceus (HE) is a well-known mushroom in traditional Chinese food and medicine. HE extracts from the fruiting body and mycelia not only exhibit immunomodulatory, antimutagenic and antitumor activity but also have neuroprotective properties. Here, we purified HE polysaccharides (HEPS), composed of two high molecular weight polysaccharides (1.7 × 10(5) Da and 1.1 × 10(5) Da), and evaluated their protective effects on amyloid beta (Aβ)-induced neurotoxicity in rat pheochromocytoma PC12 cells. HEPS were prepared and purified using a 95 % ethanol extraction method. The components of HEPS were analyzed and the molecular weights of the polysaccharides were determined using high-pressure liquid chromatography (HPLC). The neuroprotective effects of the polysaccharides were evaluated through a 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay and an MTT assay and by quantifying reactive oxygen species (ROS) and mitochondrial membrane potentials (MMP) of Aβ-induced neurotoxicity in cells. Our results showed that 250 μg/ml HEPS was harmless and promoted cell viability with 1.2 μM Aβ treatment. We observed that the free radical scavenging rate exceeded 90 % when the concentration of HEPS was higher than 1 mg/mL in cells. The HEPS decreased the production of ROS from 80 to 58 % in a dose-dependent manner. Cell pretreatment with 250 μg/mL HEPS significantly reduced Aβ-induced high MMPs from 74 to 51 % and 94 to 62 % at 24 and 48 h, respectively. Finally, 250 μg/mL of HEPS prevented Aβ-induced cell shrinkage and nuclear degradation of PC12 cells. Our results demonstrate that HEPS exhibit antioxidant and neuroprotective effects on Aβ-induced neurotoxicity in neurons.

  17. Pulsed NMR studies of crosslinking and entanglements in high molecular weight linear polydimethylsiloxanes

    International Nuclear Information System (INIS)

    Folland, R.; Charlesby, A.

    1977-01-01

    Pulsed NMR studies of proton spin relaxation are used to investigate both radiation-induced cross linking and entanglements in three high molecular weight linear polydimethylsiloxanes (Msub(w) = 26,000, 63,000 and 110,000). Particular emphasis is placed on the spin-spin relaxation since this is determined by the slower relative translational motions of the polymer chains and hence profoundly affected by the presence of intermolecular couplings such as crosslinks or entanglements. The spin-lattice relaxation times, T 1 , are determined by the fast anisotropic chain rotations and are rather insensitive to such intermolecular couplings. The spin-spin relaxation in these materials is represented by a double exponential decay involving two time constants, Tsub(2S) and Tsub(2L). The shorter component, Tsub(2S), is attributed to network material, which may be either of a dynamic form arising from temporary entanglements or of a permanent nature due to crosslinks. The concentration of entanglements depends on the initial molecular weight of the sample whereas the concentration of crosslinks is a function of the radiation dose. The longer component, Tsub(2L), is attributed to the non-network molecules. On the time scale of the NMR measurements the entanglements are shown to act in the same way as crosslinks. The variation of the relative proportions of network and non-network material with dose is shown to be accounted for by using standard gelation theory when allowance is made for the initial effective crosslink density due to entanglements. The analysis provides a value for the average molecular weight per entanglement point of 27,000 +- 1000 which is consistent with the critical molecular weight for entanglements of 29,000. The dependences of Tsub(2S) and Tsub(2L) on dose and molecular weight are also discussed in terms of the molecular motion. (author)

  18. [The high-molecular glutenins of the soft winter wheats from European countries and their relationship to the glutenin composition of the ancient and modern wheat varieties of Ukraine].

    Science.gov (United States)

    Rabinovich, S V; Fedak, G; Lukov, O

    2000-01-01

    The sources of high-quality components of HMW glutenines determining grain quality, as initial material for breeding in the conditions of Ukraine were revealed on the base of analysis of 75 literature sources data about composition of high-molecular weight (HMW) glutenin and pedigrees of 598 European wheats from 12 countries, bred in 1923-1997, including, 449 cultivars from West and 149 East Europe. Origin of these components was observed in varieties of Great Britain, France and Germany from ancient Ukrainian wheat Red Fife and it derivative spring wheats of Canada--Marquis, Garnet, Regent, Saunders, Selkirk and of USA--spring wheat Thatcher and winter wheats--Kanred and Oro--as directly as via cultivars of European countries and Australia; in wheats of East European countries from winter wheats Myronivs'ka 808 and Bezostaya 1 (derivative of Ukrainian cultivars Ukrainka and Krymka) and their descendants; in wheats of Austria and Italy--from the both genetical sources.

  19. Glutenin macropolymer: A gel formed by glutenin particles

    NARCIS (Netherlands)

    Don, C.; Lichtendonk, W.; Plijter, J.J.; Hamer, R.J.

    2003-01-01

    The quality of wheat-based foods and the processing properties of wheat flour dough are strongly related to the presence and properties of very large glutenin protein aggregates. These very large aggregates are insoluble in 1.5% (w/v) SDS and can be recovered after ultracentrifugation as a gel, the

  20. Glutenin Macropolymer: a Gel Formed by Glutenin Particles

    NARCIS (Netherlands)

    Don, C.; Lichtendonk, W.J.; Plijter, J.J.; Hamer, R.J.

    2003-01-01

    The quality of wheat-based foods and the processing properties of wheat flour dough are strongly related to the presence and properties of very large glutenin protein aggregates. These very large aggregates are insoluble in 1.5% (w/v) SDS and can be recovered after ultracentrifugation as a gel, the

  1. Biomechanical testing of new meniscal repair techniques containing ultra high-molecular weight polyethylene suture.

    Science.gov (United States)

    Barber, F Alan; Herbert, Morley A; Schroeder, F Alexander; Aziz-Jacobo, Jorge; Sutker, Michael J

    2009-09-01

    To evaluate the biomechanical characteristics of current meniscal repair techniques containing ultra high-molecular weight polyethylene (UHMWPE) suture with and without cyclic loading. Vertical longitudinal cuts made in porcine menisci were secured with a single repair device. Noncycled and cycled (500 cycles) biomechanical tests were performed on the following groups: group 1, No. 2-0 Mersilene vertical suture (Ethicon, Somerville, NJ); group 2, No. 2-0 Orthocord vertical suture (DePuy Mitek, Westwood, MA); group 3, No. 0 Ultrabraid vertical suture (Smith & Nephew Endoscopy, Andover, MA); group 4, No. 2-0 FiberWire vertical suture (Arthrex, Naples, FL); group 5, vertically oriented mattress suture by use of an Ultra FasT-Fix device (Smith & Nephew Endoscopy) with No. 0 Ultrabraid; group 6, vertically oriented mattress suture by use of a RapidLoc A2 device (DePuy Mitek) with No. 2-0 Orthocord suture; group 7, vertically oriented stitch by use of a MaxFire device with MaxBraid PE suture (Biomet Sports Medicine, Warsaw, IN); and group 8, an obliquely oriented stitch of No. 0 UHMWPE suture inserted by use of a CrossFix device (Cayenne Medical, Scottsdale, AZ). Endpoints were failure loads, failure modes, stiffness, and cyclic displacement. Mean single-pull loads were calculated for Ultra FasT-Fix (121 N), FiberWire (110 N), MaxFire (130 N), Mersilene (84 N), Orthocord (124 N), RapidLoc A2 (86 N), CrossFix (77 N), and Ultrabraid (109 N). After 500 cyclic loads, the Orthocord (222 N) repair was stronger than the others: Ultra FasT-Fix (110 N), FiberWire (117 N), MaxFire (132 N), Mersilene (89 N), RapidLoc A2 (108 N), CrossFix (95 N), and Ultrabraid (126 N) (P Fix, RapidLoc A2, and MaxFire) were comparable to the isolated UHMWPE-containing suture repairs on single-failure load testing. UHMWPE-containing suture repairs are stronger than braided polyester suture repairs, but pure UHMWPE suture (Ultrabraid) elongated more during cycling. Orthocord suture is significantly

  2. In vitro evaluation of antiviral and virucidal activity of a high molecular weight hyaluronic acid

    Directory of Open Access Journals (Sweden)

    Blasi Elisabetta

    2011-03-01

    Full Text Available Abstract Background hyaluronic acid (HA, a non-sulphated glycosaminoglycan, is present in synovial fluid, vitreous humour serum and many connective tissues. Pharmaceutical preparations of HA are used in clinical practice for wound healing, joint pain, kerato-conjunctivitis, asthma, mouth care, oesophageal-reflux, and gastritis. Moreover, it is used as a filler to counteract ageing and facial lipoatrophy. Our study aims at investigating the in vitro antiviral activity of a high molecular weight HA. Methods the MTT test was used to rule out the potential toxic effects of HA on the different cell lines used in the antiviral assays. The antiviral activity of HA against Coxsackievirus B5, Herpes Simplex Virus-1, Mumps Virus, Adenovirus-5, Influenza Virus A/H1N1, Human Herpesvirus-6, Porcine Parvovirus, Porcine Reproductive and Respiratory Syndrome Virus was assessed by virus yield assays. Results the most effective inhibition was observed against Coxsackievirus B5, with 3Log reduction of the virus yield at 4 mg/ml, and a reduction of 3.5Log and 2Log, at 2 mg/ml and 1 mg/ml, respectively: the selectivity index was 16. Mumps virus was highly inhibited too showing a reduction of 1.7Log at 1 mg/ml and 1Log at 4 mg/ml and 2 mg/ml (selectivity index = 12. The selectivity index for Influenza Virus was 12 with the highest inhibition (1Log observed at 4 mg/ml. Herpes Simplex Virus-1 and Porcine Parvovirus were mildly inhibited, whereas no antiviral activity was observed with respect to Adenovirus-5, Human Herpesvirus-6, Porcine Reproductive and Respiratory Syndrome Virus. No HA virucidal activity was ever observed against any of the viruses tested. Kinetic experiments showed that both Coxsackievirus B5 and Herpes simplex virus-1 replication were consistently inhibited, not influenced by the time of HA addition, during the virus replication cycle. Conclusions the spectrum of the antiviral activity exhibited by HA against both RNA and DNA viruses, known to have

  3. Ingestion of High Molecular Weight Carbohydrate Enhances Subsequent Repeated Maximal Power: A Randomized Controlled Trial.

    Directory of Open Access Journals (Sweden)

    Jonathan M Oliver

    Full Text Available Athletes in sports demanding repeat maximal work outputs frequently train concurrently utilizing sequential bouts of intense endurance and resistance training sessions. On a daily basis, maximal work within subsequent bouts may be limited by muscle glycogen availability. Recently, the ingestion of a unique high molecular weight (HMW carbohydrate was found to increase glycogen re-synthesis rate and enhance work output during subsequent endurance exercise, relative to low molecular weight (LMW carbohydrate ingestion. The effect of the HMW carbohydrate, however, on the performance of intense resistance exercise following prolonged-intense endurance training is unknown. Sixteen resistance trained men (23±3 years; 176.7±9.8 cm; 88.2±8.6 kg participated in a double-blind, placebo-controlled, randomized 3-way crossover design comprising a muscle-glycogen depleting cycling exercise followed by ingestion of placebo (PLA, or 1.2 g•kg•bw-1 of LMW or HMW carbohydrate solution (10% with blood sampling for 2-h post-ingestion. Thereafter, participants performed 5 sets of 10 maximal explosive repetitions of back squat (75% of 1RM. Compared to PLA, ingestion of HMW (4.9%, 90%CI 3.8%, 5.9% and LMW (1.9%, 90%CI 0.8%, 3.0% carbohydrate solutions substantially increased power output during resistance exercise, with the 3.1% (90% CI 4.3, 2.0% almost certain additional gain in power after HMW-LMW ingestion attributed to higher movement velocity after force kinematic analysis (HMW-LMW 2.5%, 90%CI 1.4, 3.7%. Both carbohydrate solutions increased post-exercise plasma glucose, glucoregulatory and gut hormones compared to PLA, but differences between carbohydrates were unclear; thus, the underlying mechanism remains to be elucidated. Ingestion of a HMW carbohydrate following prolonged intense endurance exercise provides superior benefits to movement velocity and power output during subsequent repeated maximal explosive resistance exercise. This study was registered

  4. Total and high molecular weight adiponectin have similar utility for the identification of insulin resistance

    Directory of Open Access Journals (Sweden)

    Aguilar-Salinas Carlos A

    2010-06-01

    Full Text Available Abstract Background Insulin resistance (IR and related metabolic disturbances are characterized by low levels of adiponectin. High molecular weight adiponectin (HMWA is considered the active form of adiponectin and a better marker of IR than total adiponectin. The objective of this study is to compare the utility of total adiponectin, HMWA and the HMWA/total adiponectin index (SA index for the identification of IR and related metabolic conditions. Methods A cross-sectional analysis was performed in a group of ambulatory subjects, aged 20 to 70 years, in Mexico City. Areas under the receiver operator characteristic (ROC curve for total, HMWA and the SA index were plotted for the identification of metabolic disturbances. Sensitivity and specificity, positive and negative predictive values, and accuracy for the identification of IR were calculated. Results The study included 101 men and 168 women. The areas under the ROC curve for total and HMWA for the identification of IR (0.664 vs. 0.669, P = 0.74, obesity (0.592 vs. 0.610, P = 0.32, hypertriglyceridemia (0.661 vs. 0.671, P = 0.50 and hypoalphalipoproteinemia (0.624 vs. 0.633, P = 0.58 were similar. A total adiponectin level of 8.03 μg/ml was associated with a sensitivity of 57.6%, a specificity of 65.9%, a positive predictive value of 50.0%, a negative predictive value of 72.4%, and an accuracy of 62.7% for the diagnosis of IR. The corresponding figures for a HMWA value of 4.25 μg/dl were 59.6%, 67.1%, 51.8%, 73.7% and 64.2%. The area under the ROC curve of the SA index for the identification of IR was 0.622 [95% CI 0.554-0.691], obesity 0.613 [95% CI 0.536-0.689], hypertriglyceridemia 0.616 [95% CI 0.549-0.683], and hypoalphalipoproteinemia 0.606 [95% CI 0.535-0.677]. Conclusions Total adiponectin, HMWA and the SA index had similar utility for the identification of IR and metabolic disturbances.

  5. Ultra-Fast RAFT-HDA Click Conjugation: An Efficient Route to High Molecular Weight Block Copolymers.

    Science.gov (United States)

    Inglis, Andrew J; Stenzel, Martina H; Barner-Kowollik, Christopher

    2009-11-02

    The use of the reversible addition fragmentation chain transfer-hetero Diels-Alder (RAFT-HDA) click reaction for the modular construction of block copolymers is extended to the generation of high molecular weight materials. Cyclopentadienyl end-functionalized polystyrene (PS-Cp) prepared via both atom transfer radical polymerization (ATRP) and the RAFT process are conjugated to poly(isobornyl acrylate) (PiBoA) (also prepared via RAFT polymerization) to achieve well-defined block copolymers with molecular weights ranging from 34 000 to over 100 000 g · mol(-1) and with small polydispersities (PDI HDA click chemistry can provide access to high molecular weight block copolymers in a simple and straight-forward fashion. Copyright © 2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Angioplastic necrolytic migratory erythema. Unique association of necrolytic migratory erythema, extensive angioplasia, and high molecular weight glucagon-like polypeptide

    International Nuclear Information System (INIS)

    Franchimont, C.; Pierard, G.E.; Luyckx, A.S.; Gerard, J.; Lapiere, C.M.

    1982-01-01

    A diabetic patient developed necrolytic migratory erythema with extensive angioplasia and high molecular weight glucagon-like polypeptide. There was no associated neoplasm such as glucagonoma. Lesions in the skin were studied by standard optical microscopy and by radioautography after incorporation of tritiated thymidine. Alterations in the skin begin as focal necrosis in the epidermis and in epithelial structures of adnexa, followed by marked angioplasia and a superficial and deep perivascular dermatitis

  7. Improving the Application of High Molecular Weight Biotinylated Dextran Amine for Thalamocortical Projection Tracing in the Rat.

    Science.gov (United States)

    Xu, Dongsheng; Cui, Jingjing; Wang, Jia; Zhang, Zhiyun; She, Chen; Bai, Wanzhu

    2018-04-12

    High molecular weight biotinylated dextran amine (BDA) has been used as a highly sensitive neuroanatomical tracer for many decades. Since the quality of its labeling was affected by various factors, here, we provide a refined protocol for the application of high molecular weight BDA for studying optimal neural labeling in the central nervous system. After stereotactic injection of BDA into the ventral posteromedial nucleus (VPM) of the thalamus in the rat through a delicate glass pipette, BDA was stained with fluorescent streptavidin-Alexa (AF) 594 and counterstained with fluorescent Nissl stain AF500/525. On the background of green Nissl staining, the red BDA labeling, including neuronal cell bodies and axonal terminals, was more distinctly demonstrated in the somatosensory cortex. Furthermore, double fluorescent staining for BDA and the calcium-binding protein parvalbumin (PV) was carried out to observe the correlation of BDA labeling and PV-positive interneurons in the cortical target, providing the opportunity to study the local neural circuits and their chemical characteristics. Thus, this refined method is not only suitable for visualizing high quality neural labeling with the high molecular weight BDA through reciprocal neural pathways between the thalamus and cerebral cortex, but also will permit the simultaneous demonstration of other neural markers with fluorescent histochemistry or immunochemistry.

  8. Malaria Parasite CLAG3, a Protein Linked to Nutrient Channels, Participates in High Molecular Weight Membrane-Associated Complexes in the Infected Erythrocyte.

    Directory of Open Access Journals (Sweden)

    Kayvan Zainabadi

    Full Text Available Malaria infected erythrocytes show increased permeability to a number of solutes important for parasite growth as mediated by the Plasmodial Surface Anion Channel (PSAC. The P. falciparum clag3 genes have recently been identified as key determinants of PSAC, though exactly how they contribute to channel function and whether additional host/parasite proteins are required remain unknown. To begin to answer these questions, I have taken a biochemical approach. Here I have used an epitope-tagged CLAG3 parasite to perform co-immunoprecipitation experiments using membrane fractions of infected erythrocytes. Native PAGE and mass spectrometry studies reveal that CLAG3 participate in at least three different high molecular weight complexes: a ~720kDa complex consisting of CLAG3, RHOPH2 and RHOPH3; a ~620kDa complex consisting of CLAG3 and RHOPH2; and a ~480kDa complex composed solely of CLAG3. Importantly, these complexes can be found throughout the parasite lifecycle but are absent in untransfected controls. Extracellular biotin labeling and protease susceptibility studies localize the 480kDa complex to the erythrocyte membrane. This complex, likely composed of a homo-oligomer of 160kDa CLAG3, may represent a functional subunit, possibly the pore, of PSAC.

  9. The validity of sedimentation data from high molecular weight DNA and the effects of additives on radiation-induced single-strand breakage

    International Nuclear Information System (INIS)

    Dugle, D.L.

    1979-10-01

    The optimization of many of the factors governing reproducible sedimentation behaviour of high molecular weight single-strand DNA in a particular alkaline sucrose density gradient system is described. A range of angular momenta is defined for which a constant strand breakage efficiency is required, despite a rotor speed effect which increases the measured molecular weights at decreasing rotor speeds for larger DNA molecules. The possibility is discussed that the bimodal control DNA profiles obtained after sedimentation at 11 500 rev/min (12 400 g) or less represent structural subunits of the chromatid. The random induction of single-strand DNA breaks by ionizing radiation is demonstrated by the computer-derived fits to the experimental profiles. The enhancement of single-strand break (SSB) yields in hypoxic cells by oxygen, para-nitroacetophenone (PNAP), or any of the three nitrofuran derivatives used was well correlated with increased cell killing. Furthermore, reductions in SSB yields for known hydroxyl radical (OH.) scavengers correlates with the reactivities of these compounds toward OH.. This supports the contention that some type of OH.-induced initial lesion, which may ultimately be expressed as an unrepaired or misrepaired double-strand break, constitutes a lethal event. (author)

  10. Purification of two high molecular weight proteases from rabbit reticulocyte lysate

    International Nuclear Information System (INIS)

    Hough, R.; Pratt, G.; Rechsteiner, M.

    1987-01-01

    The authors have purified two large proteases from rabbit reticulocyte lysate. The enzymes are so similar in their chromatographic behavior that each is the only significant contaminant of the other during the final stages of purification. At pH 7.8, both hydrolyze 125 I-α-casein and 4-methylcoumaryl-7-amide (MCA) derivatives with tyrosine, phenylalanine or arginine at the P 1 position. The larger, ATP-dependent enzyme degrades ubiquitin-lysozyme conjugates, but it does not degrade unmodified lysozyme. Hydrolysis of Suc-Leu-Leu-Val-Tyr-MCA by this enzyme is also stimulated two-fold in the presence of ATP. The protease has a molecular weight of 950,000 based on sedimentation, gel filtration and non-denaturing PAGE. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the protease is composed of a number of subunits with molecular masses between 32 and 110 kDa. Densitometric analysis showed equivalent amounts of the two larger chains, and the presence of one copy of each in the native enzyme would be consistent with an M/sub r/ of 950,000. The smaller protease has a molecular weight of 700,000 and is composed of 8 to 10 subunits ranging from 21,000 to 32,000. It cleaves ubiquitin-lysozyme conjugates only slightly, and hydrolysis of conjugates or fluorogenic peptide substrates is not stimulated by ATP. This protease appears similar, if not identical, to the multicatalytic protease complex first purified by Wilk and Orlowski

  11. Degradation and mineralization of high-molecular-weight polycyclic aromatic hydrocarbons by defined fungal-bacterial cocultures

    International Nuclear Information System (INIS)

    Boonchan, S.; Britz, M.L.; Stanley, G.A.

    2000-01-01

    This study investigated the biodegradation of high-molecular-weight polycyclic aromatic hydrocarbons (PAHs) in liquid media and soil by bacteria (Stenotrophomonas maltophilia VUN 10,010 and bacterial consortium VUN 10,009) and a fungus (Penicillium janthinellum VUO 10,201) that were isolated from separate creosote- and manufactured-gas plant-contaminated soils. The bacteria could use pyrene as their sole carbon and energy source in a basal salts medium (BSM) and mineralized significant amounts of benzo[a]pyrene cometabolically when pyrene was also present in BSM. P. janthinellum VUO 10,201 could not utilize any high-molecular-weight PAH as sole carbon and energy source but could partially degrade these if cultured in a nutrient broth. Although small amounts of chrysene, benz[a]pyrene, and dibenz[a,h]anthracene were degraded by axenic cultures of these isolates in BSM containing a single PAH, such conditions did not support significant microbial growth or PAH mineralization. However, significant degradation of, and microbial growth on, pyrene, chrysene, benz[a]anthracene, benzo[a]pyrene, and dibenz[a,h]anthracene, each as a single PAH in BSM, occurred when P. janthinellum VUO 10,201 and either bacterial consortium VUN 10,009 or S. maltophilia VUN 10,010 were combined in the one culture, i.e., fungal-bacterial cocultures: 25% of the benzo[a]pyrene was mineralized to CO 2 by these cocultures over 49 days, accompanied by transient accumulation and disappearance of intermediates detected by high-pressure liquid chromatography. Inoculation of fungal-bacterial cocultures into PAH-contaminated soil resulted in significantly improved degradation of high-molecular-weight PAHs, benzo[a]pyrene mineralization, and reduction in the mutagenicity of organic soil extracts, compared with the indigenous microbes and soil amended with only axenic inocula

  12. Effect of electron beam radiation on the structure and mechanical properties of ultra high molecular weight polyethylene fibers

    International Nuclear Information System (INIS)

    Li Shujun; Sun Weijun; Liu Xiuju; Gao Yongzhong; Li Huisheng

    1998-01-01

    Ultra high molecular weight polyethylene fibers have been crosslinked by electron beam. The structure and mechanical properties of them have been investigated in different irradiation atmospheres. The obtained results show that the gel content and crosslinking density increase with the increase of dose, the swelling ratio and average molecular weight of crosslinked net decrease with the increase of dose, the tensile strength and failure elongation decrease with the increase of dose, the tensile modulus increases with the increase of dose. When the samples are irradiated in air, vacuum and acetylene atmospheres, the effect of irradiation in acetylene atmosphere is best

  13. Rheological Link Between Polymer Melts with a High Molecular Weight Tail and Enhanced Formation of Shish-Kebabs

    DEFF Research Database (Denmark)

    Wingstrand, Sara Lindeblad; Shen, Bo; Kornfield, Julie A.

    2017-01-01

    Presence of an ultra high molecular weight (UHMw) fraction in flowingpolymer melts is known to facilitate formation of oriented crystalline structures significantly. The UHMw fraction manifests itself as a minor tail in the molar mass distribution and is hardly detectable in the canonical...... a clear increase in extensional stress that is directly correlated with the crystalline orientation of the quenched samples. Extensional rheology, particularly, in combination with linear creep measurements, thus, enables the conformational evolution of the UHMw-tail to be studied and linked...

  14. Kinetics of low temperature polyester dyeing with high molecular weight disperse dyes by solvent microemulsion and agrosourced auxiliaries

    OpenAIRE

    Radei, Shahram; Carrión-Fité, Francisco Javier; Ardanuy Raso, Mònica; Canal Arias, José Ma

    2018-01-01

    This work focused on the evaluation of the kinetics of dyeing polyester fabrics with high molecular weight disperse dyes, at low temperature by solvent microemulsion. This study also compared the effect of two non-toxic agro-sourced auxiliaries (o-vanillin and coumarin) using a non-toxic organic solvent. A dyeing bath consisting of a micro-emulsion system involving a small proportion of n-butyl acetate was used, and the kinetics of dyeing were analysed at four temperatures (83, 90, 95 and 100...

  15. Kinetics of Low Temperature Polyester Dyeing with High Molecular Weight Disperse Dyes by Solvent Microemulsion and AgroSourced Auxiliaries

    OpenAIRE

    Shahram Radei; F. Javier Carrión-Fité; Mònica Ardanuy; José María Canal

    2018-01-01

    This work focused on the evaluation of the kinetics of dyeing polyester fabrics with high molecular weight disperse dyes, at low temperature by solvent microemulsion. This study also compared the effect of two non-toxic agro-sourced auxiliaries (o-vanillin and coumarin) using a non-toxic organic solvent. A dyeing bath consisting of a micro-emulsion system involving a small proportion of n-butyl acetate was used, and the kinetics of dyeing were analysed at four temperatures (83, 90, 95 and 100...

  16. Identification of unprecedented anticancer properties of high molecular weight biomacromolecular complex containing bovine lactoferrin (HMW-bLf.

    Directory of Open Access Journals (Sweden)

    Fawzi Ebrahim

    Full Text Available With the successful clinical trials, multifunctional glycoprotein bovine lactoferrin is gaining attention as a safe nutraceutical and biologic drug targeting cancer, chronic-inflammatory, viral and microbial diseases. Interestingly, recent findings that human lactoferrin oligomerizes under simulated physiological conditions signify the possible role of oligomerization in the multifunctional activities of lactoferrin molecule during infections and in disease targeting signaling pathways. Here we report the purification and physicochemical characterization of high molecular weight biomacromolecular complex containing bovine lactoferrin (≥250 kDa, from bovine colostrum, a naturally enriched source of lactoferrin. It showed structural similarities to native monomeric iron free (Apo lactoferrin (∼78-80 kDa, retained anti-bovine lactoferrin antibody specific binding and displayed potential receptor binding properties when tested for cellular internalization. It further displayed higher thermal stability and better resistance to gut enzyme digestion than native bLf monomer. High molecular weight bovine lactoferrin was functionally bioactive and inhibited significantly the cell proliferation (p<0.01 of human breast and colon carcinoma derived cells. It induced significantly higher cancer cell death (apoptosis and cytotoxicity in a dose-dependent manner in cancer cells than the normal intestinal cells. Upon cellular internalization, it led to the up-regulation of caspase-3 expression and degradation of actin. In order to identify the cutting edge future potential of this bio-macromolecule in medicine over the monomer, its in-depth structural and functional properties need to be investigated further.

  17. Use of I-131 labeled, murine Fab against a high molecular weight antigen of human melanoma: Preliminary experience

    International Nuclear Information System (INIS)

    Larson, S.M.; Carrasquillo, J.A.; McGuffin, R.W.

    1985-01-01

    High molecular-weight antigen (HMWA) is tumor-associated proteoglycan of human malignant melanoma. I-131 labeled Fab fragments of these specific antibodies were used for preliminary feasibility studies for radioimmunodetection and therapy of human subjects who had inoperable metastatic melanoma. Ten patients received tracer doses of I-131 (anti-HMWA) Fab. All patients (8/8) who had melanoma lesions greater than 1 cm by correlative diagnosis methods had one or more lesions that had localization to tumor of the radiolabelled Fab. In all, 17 of 23 (74%) documented metastases were seen. Two patients who had avid uptake received potentially radiotherapeutic doses. For both of these patients, whole imaging studies showed that the localization of the high dose I-131 Fab was predominantly in tumor. On whole body images, the anti-Fab HMWA appears to be more tumor selective than Fab preparations that target the p97 antigen for melanoma, and there is less uptake in liver

  18. Ultra-High Molecular Weight Polyethylene: Influence of the Chemical, Physical and Mechanical Properties on the Wear Behavior. A Review

    Directory of Open Access Journals (Sweden)

    Pierangiola Bracco

    2017-07-01

    Full Text Available Ultra-high molecular weight polyethylene (UHMWPE is the most common bearing material in total joint arthroplasty due to its unique combination of superior mechanical properties and wear resistance over other polymers. A great deal of research in recent decades has focused on further improving its performances, in order to provide durable implants in young and active patients. From “historical”, gamma-air sterilized polyethylenes, to the so-called first and second generation of highly crosslinked materials, a variety of different formulations have progressively appeared in the market. This paper reviews the structure–properties relationship of these materials, with a particular emphasis on the in vitro and in vivo wear performances, through an analysis of the existing literature.

  19. Physico-chemical characterization of polyethylene of ultra high molecular weight modified with gamma irradiation and heavy ions

    International Nuclear Information System (INIS)

    Lagarde, M; Del Grosso, M; Fasce, D; Dommarco, R; Laino, S; Fasce, L.A

    2012-01-01

    The ultra high molecular weight polyethylene (UHMWPE) is a biomaterial widely used in total joint replacement. In this work, the effect of two different irradiation techniques on UHMWPE is analyzed. One technique involves gamma irradiation (γ) followed by a thermal treatment, thus modifying the material bulk. The other implies swift heavy ion irradiation (SHI), which have an effect only on the near surface layers. The surface nanomechanical properties are evaluated from depth sensing indentation experiments, while changes in crystallinity and chemical structure are determined by DSC and Raman spectroscopy. The results show that even when both techniques are able to improve the UHMWPE wear behavior, the effect on other mechanical properties and molecular structure modification is different. The γ irradiated sample exhibits lower crystallinity, hardness and modulus than the pristine UHMWPE, while the SHI irradiated sample exhibits higher crystallinity and enhanced mechanical properties than the later

  20. A Tribological Assessment of Ultra High Molecular Weight Polyethylene Types GUR 1020 and GUR 1050 for Orthopedic Applications

    Directory of Open Access Journals (Sweden)

    Benjamin J. Hunt

    2016-06-01

    Full Text Available The wear properties of biomaterials have been demonstrated to have a high importance within orthopedic bearing surfaces. This study performed a comparison of the wear between the two main grades of Ultra High Molecular Weight Polyethylene types GUR 1020 and GUR 1050 articulating against Cobalt Chromium. Such a high capacity wear comparison has not been reported elsewhere in the scientific literature. Under an identical testing protocol it was found that GUR 1020 had a wear factor of 3.92 ± 0.55 × 10 − 6 ( mm 3 / Nm and GUR 1050 had a wear factor of 3.64 ± 0.39 × 10 − 6 ( mm 3 / Nm , with a non-statistical significant difference of p = 0.052. These wear factors correlate closely with those observed from other screening wear studies and explant analysis.

  1. Convenient preparation of high molecular weight poly(dimethylsiloxane using thermally latent NHC-catalysis: a structure-activity correlation

    Directory of Open Access Journals (Sweden)

    Stefan Naumann

    2015-11-01

    Full Text Available The polymerization of octamethylcyclotetrasiloxane (D4 is investigated using several five-, six- and seven-membered N-heterocyclic carbenes (NHCs. The catalysts are delivered in situ from thermally susceptible CO2 adducts. It is demonstrated that the polymerization can be triggered from a latent state by mild heating, using the highly nucleophilic 1,3,4,5-tetramethylimidazol-2-ylidene as organocatalyst. This way, high molecular weight PDMS is prepared (up to >400 000 g/mol, 1.6 ÐM 95%, using low catalyst loadings (0.2–0.1 mol %. Furthermore, the results suggest that a nucleophilic, zwitterionic mechanism is in operation, in preference to purely anionic polymerization.

  2. Relationship between the visceral fat area and adipocytokine in hemodialysis patients. Usefulness of high molecular weight adiponectin-leptin ratio

    International Nuclear Information System (INIS)

    Tsushima, Megumi; Terayama, Yuriko; Tsutaya, Chikako; Momose, Akishi; Funyu, Tomihisa; Ohyama, Chikara; Hada, Ryukichi

    2007-01-01

    Adiponectin and leptin, adipose-specific secretory proteins, are associated with obesity. It has been reported that the plasma adiponectin and leptin concentrations were both higher in women than in men, and were markedly increased among hemodialysis patients in comparison with healthy controls. This study was conduced in 104 hemodialysis patients and 35 healthy controls. To obtain a novel indicator for the visceral fat area in hemodialysis patients, we measured total adiponectin (t-Adipo), high molecular weight adiponectin (h-Adipo) and leptin (Lep) in serum and evaluated visceral and subcutaneous fat areas using computed tomography. In addition, we calculated the percentage of the high molecular weight form per total adiponectin (h-Adipo/t-Adipo) and the ratio of adiponectin to leptin (Adipo/Lep ratio). Hemodialysis patients were divided into four groups based on their visceral fat area level (under 40 cm 2 ; group I, 40-70 cm 2 ; group II, 70-100 cm 2 ; group III, 100 cm 2 or more; group IV). T-Adipo, h-Adipo, leptin and h-Adipo/t-Adipo were significantly higher in women than in men. T-Adipo, h-Adipo and h-Adipo/t-Adipo were lower in group IV than in group I. Leptin were higher in group III and IV than in group I. The T-Adipo/Lep ratio and h-Adipo/Lep ratio were lower in group III and IV than in group I. Especially, the h-Adipo/Lep ratio in group IV was significantly decreased compared with that in group III. By simple regression analysis of the correlation between visceral fat area and each parameter of adipocytokine, the highest correlation coefficients were seen with the log transformed h-Adipo/Lep ratio. Log transformed h-Adipo/Lep ratio were inversely related to visceral fat area for men (y=-36.16 x +75.8, r=-0.558, p<0.001) and for women (y=-36.07 x +68.2, r=-0.725, p<0.001). The two regression lines between-gender difference were equivalent in slope. We conclude that h-Adipo/Lep ratio may be a marker for evaluating visceral fat area in hemodialysis

  3. Comparative proteome analysis of glutenin synthesis and accumulation in developing grains between superior and poor quality bread wheat cultivars.

    Science.gov (United States)

    Liu, Wan; Zhang, Yanzhen; Gao, Xuan; Wang, Ke; Wang, Shunli; Zhang, Yong; He, Zhonghu; Ma, Wujun; Yan, Yueming

    2012-01-15

    Wheat glutenins are the major determinants of wheat quality. In this study, grains at the development stage from three wheat cultivars (Jimai 20, Jin 411 and Zhoumai 16) with different bread-making quality were harvested based on thermal times from 150 °C(d) to 750 °C(d) , and were used to investigate glutenin accumulation patterns and their relationships with wheat quality. High and low molecular weight glutenin subunits (HMW-GSs and LMW-GSs) were synthesised concurrently. No obvious correlations between HMW/LMW glutenin ratios and dough property were observed. Accumulation levels of HMW-GSs and LMW-GSs as well as 1Bx13 + 1By16 and 1Dx4 + 1Dy12 subunits were higher in superior gluten quality cultivar Jimain 20 than in poor quality cultivar Jing 411 and Zhoumai 16. According to the results of two-dimensional gel electrophoresis, six types of accumulation patterns in LMW-GSs were identified and classified. The possible relationships between individual LMW-GSs and gluten quality were established. The high accumulation level of HMW-GSs and LMW-GSs as well as 1Bx13 + 1By16 and 1Dx4 + 1Dy12 subunits contributed to the superior gluten quality of Jimai 20. Two highly expressed and 16 specifically expressed LMW glutenin subunits in Jimain 20 had positive effects on dough quality, while 17 specifically expressed subunits in Zhoumai 16 and Jing 411 appeared to have negative effects on gluten quality. Copyright © 2011 Society of Chemical Industry.

  4. Relationship between Oversulfation and Conformation of Low and High Molecular Weight Fucoidans and Evaluation of Their in Vitro Anticancer Activity

    Directory of Open Access Journals (Sweden)

    Myoung Lae Cho

    2010-12-01

    Full Text Available Low and high molecular weight fucoidans (F5-30K and F>30K were chemically modified through the addition of sulfate groups, and the effect of oversulfation on the in vitro anticancer activity was investigated. After the addition of sulfate groups, a considerable increase of 35.5 to 56.8% was observed in the sulfate content of the F5-30K fraction, while the sulfate content of the F>30K fraction increased to a lesser extent (from 31.7 to 41.2%. Significant differences in anticancer activity were observed between the oversulfated F5–30K and F>30K fractions, with activities of 37.3–68.0% and 20.6–35.8%, respectively. This variation in the anticancer activity of oversulfated fucoidan derivatives was likely due to differences in their sulfate content. The results suggest that the molecular conformation of these molecules is closely related to the extent of sulfation in the fucan backbones and that the sulfates are preferably substituted when the fucoidan polymers are in a loose molecular conformation.

  5. Distinct high molecular weight organic compound (HMW-OC) types in aerosol particles collected at a coastal urban site

    Science.gov (United States)

    Dall'Osto, M.; Healy, R. M.; Wenger, J. C.; O'Dowd, C.; Ovadnevaite, J.; Ceburnis, D.; Harrison, Roy M.; Beddows, D. C. S.

    2017-12-01

    Organic oligomers were discovered in laboratory-generated atmospheric aerosol over a decade ago. However, evidence for the presence of oligomers in ambient aerosols is scarce and mechanisms for their formation have yet to be fully elucidated. In this work, three unique aerosol particle types internally mixed with High molecular weight organic compounds (HMW-OC) species - likely oligomers - were detected in ambient air using single particle Aerosol Time-Of-Flight Mass Spectrometry (ATOFMS) in Cork (Ireland) during winter 2009. These particle types can be described as follows: (1) HMW-OCs rich in organic nitrogen - possibly containing nitrocatechols and nitroguaiacols - originating from primary emissions of biomass burning particles during evening times; (2) HMW-OCs internally mixed with nitric acid, occurring in stagnant conditions during night time; and (3) HMW-OCs internally mixed with sea salt, likely formed via photochemical reactions during day time. The study exemplifies the power of methodologies capable of monitoring the simultaneous formation of organic and inorganic particle-phase reaction products. Primary emissions and atmospheric aging of different types of HMW-OC contributes to aerosol with a range of acidity, hygroscopic and optical properties, which can have different impacts on climate and health.

  6. Biodegradation of low and high molecular weight hydrocarbons in petroleum refinery wastewater by a thermophilic bacterial consortium.

    Science.gov (United States)

    Pugazhendi, Arulazhagan; Abbad Wazin, Hadeel; Qari, Huda; Basahi, Jalal Mohammad Al-Badry; Godon, Jean Jacques; Dhavamani, Jeyakumar

    2017-10-01

    Clean-up of contaminated wastewater remains to be a major challenge in petroleum refinery. Here, we describe the capacity of a bacterial consortium enriched from crude oil drilling site in Al-Khobar, Saudi Arabia, to utilize polycyclic aromatic hydrocarbons (PAHs) as sole carbon source at 60°C. The consortium reduced low molecular weight (LMW; naphthalene, phenanthrene, fluorene and anthracene) and high molecular weight (HMW; pyrene, benzo(e)pyrene and benzo(k)fluoranthene) PAH loads of up to 1.5 g/L with removal efficiencies of 90% and 80% within 10 days. PAH biodegradation was verified by the presence of PAH metabolites and evolution of carbon dioxide (90 ± 3%). Biodegradation led to a reduction of the surface tension to 34 ± 1 mN/m thus suggesting biosurfactant production by the consortium. Phylogenetic analysis of the consortium revealed the presence of the thermophilic PAH degrader Pseudomonas aeruginosa strain CEES1 (KU664514) and Bacillus thermosaudia (KU664515) strain CEES2. The consortium was further found to treat petroleum wastewater in continuous stirred tank reactor with 96 ± 2% chemical oxygen demand removal and complete PAH degradation in 24 days.

  7. The effect of gamma irradiation and shelf aging in air on the oxidation of ultra-high molecular weight polyethylene

    International Nuclear Information System (INIS)

    Al-Ma'adeed, M.A.; Al-Qaradawi, I.Y.; Madi, N.; Al-Thani, N.J.

    2006-01-01

    This study has investigated the effect of shelf aging, for up to one year in air, on the properties of gamma-irradiated ultra-high molecular weight polyethylene (UHMWPE). A variety of techniques were used to characterize the properties of treated samples. Differential scanning calorimetery (DSC) was used to characterize the morphology. The extent of cross-linking in a polymer network was detected by swelling measurements. The durometer hardness test was used to measure the relative hardness of this material, and changes in density were also measured. Results from all these measurements were combined to explain the changes in the microstructure of the aged, irradiated UHMWPE. This study shows that crystallinity is increased with radiation dose and with aging due to chain scission, which leads to a reduction in the molecular weight of the material. This allows the chains to rearrange to form crystalline regions. Positron annihilation lifetime spectroscopy confirms these conclusions. Fractional free volumes have been deduced from lifetime parameters, which correlate with the data obtained by the other techniques

  8. Kinetics of Low Temperature Polyester Dyeing with High Molecular Weight Disperse Dyes by Solvent Microemulsion and AgroSourced Auxiliaries

    Directory of Open Access Journals (Sweden)

    Shahram Radei

    2018-02-01

    Full Text Available This work focused on the evaluation of the kinetics of dyeing polyester fabrics with high molecular weight disperse dyes, at low temperature by solvent microemulsion. This study also compared the effect of two non-toxic agro-sourced auxiliaries (o-vanillin and coumarin using a non-toxic organic solvent. A dyeing bath consisting of a micro-emulsion system involving a small proportion of n-butyl acetate was used, and the kinetics of dyeing were analysed at four temperatures (83, 90, 95 and 100 °C. Moreover, the dyeing rate constants, correlation coefficient and activation energies were proposed for this system. It was found that o-vanillin yielded higher dye absorption levels than coumarin, leading to exhaustions of 88% and 87% for Disperse Red 167 and Disperse Blue 79, respectively. K/S values of dyed polyester were also found to be higher for dye baths containing o-vanillin with respect to the ones with coumarin. In terms of hot pressing fastness and wash fastness, generally no adverse influence on fastness properties was reported, while o-vanillin showed slightly better results compared to coumarin.

  9. Development of solvent-free offset ink using vegetable oil esters and high molecular-weight resin.

    Science.gov (United States)

    Park, Jung Min; Kim, Young Han; Kim, Sung Bin

    2013-01-01

    In the development of solvent-free offset ink, the roles of resin molecular weight and used solvent on the ink performance were evaluated by examining the relationship between the various properties of resin and solvent and print quality. To find the best performing resin, the soy-oil fatty acid methyl ester (FAME) was applied to the five modified-phenolic resins having different molecular weights. It is found from the experimental results that the ink made of higher molecular weight and better solubility resin gives better printability and print quality. It is because larger molecular weight resin with better solubility gives higher rate of ink transfer. From the ink application of different esters to high molecular weight resin, the best printing performance was yielded from the soy-oil fatty acid butyl ester (FABE). It is due to its high kinematic viscosity resulting in the smallest change of ink transfer weight upon multiple number of printing, which improves the stability of ink quality.

  10. Highly hydrophilic ultra-high molecular weight polyethylene powder and film prepared by radiation grafting of acrylic acid

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Honglong [Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Xu, Lu; Li, Rong [Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800 (China); Pang, Lijuan [Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Hu, Jiangtao; Wang, Mouhua [Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800 (China); Wu, Guozhong, E-mail: wuguozhong@sinap.ac.cn [Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800 (China)

    2016-09-30

    Highlights: • Hydrophilic UHMWPE powder and film were obtained by γ-ray pre-irradiation grafting of AA. • A low concentration of AA solution was used for surface modification of UHMWPE. • A small grafting yield of AA sufficiently improved hydrophilicity of UHMWPE powder and film. - Abstract: The surface properties of ultra-high molecular weight polyethylene (UHMWPE) are very important for its use in engineering or composites. In this work, hydrophilic UHMWPE powder and film were prepared by γ-ray pre-irradiation grafting of acrylic acid (AA) and further neutralization with sodium hydroxide solution. Variations in the chemical structure, grafting yield and hydrophilicity were investigated and compared. FT-IR and XPS analysis results showed that AA was successfully grafted onto UHMWPE powder and film; the powder was more suitable for the grafting reaction in 1 wt% AA solution than the film. Given a dose of 300 kGy, the grafting yield of AA was ∼5.7% for the powder but ∼0.8% for the film under identical conditions. Radiation grafting of a small amount of AA significantly improved the hydrophilicity of UHMWPE. The water contact angle of the UHMWPE-g-PAA powder with a grafting yield of AA at ∼5.7% decreased from 110.2° to 68.2°. Moreover, the grafting powder (UHMWPE-g-PAA) exhibited good dispersion ability in water.

  11. Spinal motor neuron neuroaxonal spheroids in chronic aluminum neurotoxicity contain phosphatase-resistant high molecular weight neurofilament (NFH).

    Science.gov (United States)

    Gaytan-Garcia, S; Kim, H; Strong, M J

    1996-04-15

    It has previously been shown that a single intracisternal inoculum of AlCl3 in young adult New Zealand white rabbits will induce a dose-dependent phosphatase resistance of high molecular weight neurofilament protein (NFH) that is proportionate to the extent of neurofilamentous inclusion formation (Strong and Jakowec, 1994). To determine if the potential for dissolution of aluminum-induced neurofilamentous inclusions was dependent on the degree of NFH phosphatase resistance, we have examined NFH phosphatase sensitivity in a reversible chronic model of aluminum neurotoxicity. Rabbits receiving repeated intracisternal inoculums of 100 microgram AlCl3 at 28 day intervals until day 267 develop spinal motor neuron perikaryal and neuroaxonal neurofilamentous aggregates in a stereotypic, dose-dependent fashion. In the rabbits receiving inoculums until day 156 with survival until day 267 without further aluminum exposure, neuroaxonal spheroids remained prominent while perikaryal inclusions largely resolved. Immunoreactivity to a monoclonal antibody recognizing phosphorylated NFH (SMI 31) was abolished in perikaryal aggregates at each time interval by dephosphorylation with bovine alkaline phosphatase. However, neuroaxonal spheroids maintained their immunoreactivity. Using time-course dephosphorylation studies of spinal cord homogenates, we observed a significant reduction in the rate of dephosphorylation of NFH following 267 days of AlCl3 exposure (P < 0.05). These observations suggest that neuroaxonal spheroids contain phosphatase-resistant NFH isoforms and that the potential for resolution of intraneuronal neurofilamentous inclusions correlates with the susceptibility of NF within these inclusions to enzymatic dephosphorylation.

  12. The ratio of high-molecular weight adiponectin and total adiponectin differs in preterm and term infants.

    Science.gov (United States)

    Yoshida, Tomohide; Nagasaki, Hiraku; Asato, Yoshihide; Ohta, Takao

    2009-05-01

    Adiponectin consists of three subspecies (high-, middle- and low-molecular weight adiponectin). Among these, high-molecular weight adiponectin (H-adn) is suggested to be an active form of this protein. To assess the relationship between H-adn and postnatal growth in preterm infants (PIs), serum H-adn and total adiponectin (T-adn) were measured in 46 PIs at birth and at corrected term, and 26 term infants (TI) at birth. T-adn and H-adn concentrations, and the ratio of H-adn to T-adn (H/T-adn) were significantly greater in TI and PI at corrected term than in PI at birth (p adn and H-adn concentrations in PI at corrected term were similar to those in TI, but H/T-adn in PI at corrected term was less than that in TI (p adn and serum concentrations of T- and H-adn in PI at corrected term were different from those in TI. These data suggest that quality of early postnatal growth in PIs is different from that in normally developed TI. Postnatal growth accompanying adipose tissue similar to TI may be important for PI to prevent future development of cardiovascular disease.

  13. Using microwave heating to improve the desorption efficiency of high molecular weight VOC from beaded activated carbon.

    Science.gov (United States)

    Fayaz, Mohammadreza; Shariaty, Pooya; Atkinson, John D; Hashisho, Zaher; Phillips, John H; Anderson, James E; Nichols, Mark

    2015-04-07

    Incomplete regeneration of activated carbon loaded with organic compounds results in heel build-up that reduces the useful life of the adsorbent. In this study, microwave heating was tested as a regeneration method for beaded activated carbon (BAC) loaded with n-dodecane, a high molecular weight volatile organic compound. Energy consumption and desorption efficiency for microwave-heating regeneration were compared with conductive-heating regeneration. The minimum energy needed to completely regenerate the adsorbent (100% desorption efficiency) using microwave regeneration was 6% of that needed with conductive heating regeneration, owing to more rapid heating rates and lower heat loss. Analyses of adsorbent pore size distribution and surface chemistry confirmed that neither heating method altered the physical/chemical properties of the BAC. Additionally, gas chromatography (with flame ionization detector) confirmed that neither regeneration method detectably altered the adsorbate composition during desorption. By demonstrating improvements in energy consumption and desorption efficiency and showing stable adsorbate and adsorbent properties, this paper suggests that microwave heating is an attractive method for activated carbon regeneration particularly when high-affinity VOC adsorbates are present.

  14. Black Tea High-Molecular-Weight Polyphenol-Rich Fraction Promotes Hypertrophy during Functional Overload in Mice

    Directory of Open Access Journals (Sweden)

    Yuki Aoki

    2017-03-01

    Full Text Available Mitochondria activation factor (MAF is a high-molecular-weight polyphenol extracted from black tea that stimulates training-induced 5′ adenosine monophosphate-activated protein kinase (AMPK activation and improves endurance capacity. Originally, MAF was purified from black tea using butanol and acetone, making it unsuitable for food preparation. Hence, we extracted a MAF-rich sample “E80” from black tea, using ethanol and water only. Here, we examined the effects of E80 on resistance training. Eight-week old C57BL/6 mice were fed with a normal diet or a diet containing 0.5% E80 for 4, 7 and 14 days under conditions of functional overload. It was found that E80 administration promoted overload-induced hypertrophy and induced phosphorylation of the Akt/mammalian target of rapamycin (mTOR pathway proteins, such as Akt, P70 ribosomal protein S6 kinase (p70S6K, and S6 in the plantaris muscle. Therefore, functional overload and E80 administration accelerated mTOR signaling and increased protein synthesis in the muscle, thereby inducing hypertrophy.

  15. Ionizing radiation effect study by electron beam on ultra high molecular weight polyethylene virgin and recycled industrial

    International Nuclear Information System (INIS)

    Rosario, Salmo Cordeiro do

    2006-01-01

    Ultra High Molecular Weight Polyethylene (UHMWPE) is an engineering plastic which has several applications, chiefly, in specific areas of the industry and medicine. UHMWPE can be even for other applications such as: port fenders, current guide, bucket coating, silos and gutters, plugs, pulleys and surgical prosthesis. This range of applications is due to the excellent technical characteristics that this material owns, such as; high resistance to wear, high resistance to impact, anti-adherence, non toxic, excellent chemical resistance, low specific weight, easy mill processing, and high resistance to fatigue. The UHMWPE type used in this work were UTEC 3041 and UTEC 6541 of the Braskem. The recycling process of UHMWPE raised much interest, because the utilization of this raw material grew over 600% in the last decade, becoming one of the most used engineering plastics for attainment of mill processed parts after polyamide. As the utilization of this polymer in the manufacturing of parts for machinery has grown, its waste is very big, because the rest of this material is thrown out, usually not being reused. The goal of this work is to recycle the UHMWPE UTEC 3041 and study the properties of this recycled and virgin material and compare the results between both with these materials submitted to different radiation dose. (author)

  16. High molecular weight hyaluronic acid increases the differentiation potential of the murine chondrocytic ATDC5 cell line.

    Science.gov (United States)

    Sato, Eiichi; Ando, Takashi; Ichikawa, Jiro; Okita, Genki; Sato, Nobutaka; Wako, Masanori; Ohba, Tetsuro; Ochiai, Satoshi; Hagino, Tetsuo; Jacobson, Richard; Haro, Hirotaka

    2014-12-01

    Osteoarthritis (OA) is a group of common, chronic, and painful inflammatory joint diseases. One important finding in OA patients is a remarkable decrease in the molecular weight of hyaluronic acid (HA) in the synovial fluid of affected joints. Therapeutic HA is available to patients in most parts of the world as a viscosupplementation product for the treatment of OA. Previous clinical reports show that high molecular weight HA (HMWHA) more effectively relieves pain than low molecular weight HA (LMWHA). However, the mechanism behind this finding remains unclear. In this study, we investigated whether a LMWHA (Low-0.9 MDa) and two types of HMWHA (High-1.9 MDa and 6 MDa) differentially affected chondroregulatory action. We tested this using ATDC5 cell, a murine chondrocytic cell line widely used in culture systems to study chondrogenic differentiation. We found that HMWHA, especially hylan G-F 20 (High-6 MDa), significantly induced aggrecan and proteoglycan accumulation, nodule formation, and mRNA expression of chondrogenic differentiation markers in a time- and dose-dependent manner. In addition, we showed that HMWHA prevented TNF-α induced inhibition of chondrogenic differentiation, with no effect on cell proliferation or viability. These results reveal that HMWHA significantly promotes chondrogenic differentiation of ATDC5 cells in vitro, and suggest that HMWHA plays a significant chondroregulatory role in vivo. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  17. New titanium and titanium/hydroxyapatite coatings on ultra-high-molecular-weight polyethylene-in vitro osteoblastic performance

    International Nuclear Information System (INIS)

    Silva, M A; Lopes, M A; Santos, J D; Fernandes, M H; Gomes, P S; Vila, M; Silva, R F

    2010-01-01

    The development of optimized hip joint materials is one of the most challenging opportunities in prosthetic technologies. In current approaches, ultra-high-molecular-weight polyethylene (UHMWPE) has been a favorite material for the acetabular component and, regarding the cementless technique, several coating options may be considered to contain and stabilize bearing surfaces and establish an improved interface with bone. In this work, newly developed constructs of UHMWPE coated with either commercially pure titanium (cpTi-UHMWPE), by DC magnetron sputtering, or with commercially pure titanium and hydroxyapatite (cpTi/HA-UHMWPE), by DC/RF magnetron co-sputtering, have been prepared and biologically characterized with human bone marrow-derived osteoblastic cultures. The cpTi-UHMWPE samples allowed a high cell growth and the expression of the complete osteoblastic phenotype, with high alkaline phosphatase activity, expression of osteogenic-associated genes and evident cell-mediated mineralization of the extracellular matrix. In comparison, the cpTi/HA-UHMWPE samples reported lower cell proliferation but earlier cell-mediated matrix mineralization. Accordingly, these newly developed systems may be suitable candidates to improve the osteointegration process in arthroplastic devices; nevertheless, further biological evaluation should be conducted.

  18. Ring-opening polymerization of 19-electron [2]cobaltocenophanes: a route to high-molecular-weight, water-soluble polycobaltocenium polyelectrolytes.

    Science.gov (United States)

    Mayer, Ulrich F J; Gilroy, Joe B; O'Hare, Dermot; Manners, Ian

    2009-08-05

    Water-soluble, high-molecular-weight polycobaltocenium polyelectrolytes have been prepared by ring-opening polymerization (ROP) techniques. Anionic polymerization of a strained 19-electron dicarba[2]cobaltocenophane followed by oxidation in the presence of ammonium chloride resulted in the formation of oligomers with up to nine repeat units. Thermal ROP of dicarba[2]cobaltocenophane followed by oxidation in the presence of ammonium nitrate resulted in the formation of high-molecular-weight polycobaltocenium nitrate, a redox-active cobalt-containing polyelectrolyte.

  19. The origin and early development of wheat glutenin particles

    NARCIS (Netherlands)

    Herpen, van T.W.J.M.; Cordewener, J.H.G.; Klok, H.J.; Freeman, J.; America, A.H.P.; Bosch, H.J.; Smulders, M.J.M.; Gilissen, L.J.W.J.; Shewry, P.R.; Hamer, R.J.

    2008-01-01

    Breadmaking quality is strongly related to the glutenin macropolymer (GMP) fraction. Don and co-workers [Don, C., Lichtendonk, W.J., Plijter, J.J., Hamer, R.J., 2003a. Glutenin macropolymer: a gel formed by particles. Journal of Cereal Science 37, 1¿7] showed that GMP consists of spherical glutenin

  20. Resilient Amorphous Networks Prepared by Photo-Crosslinking High-Molecular-Weight D,L-Lactide and Trimethylene Carbonate Macromers: Mechanical Properties and Shape-Memory Behavior

    NARCIS (Netherlands)

    Sharifi, Shahriar; Grijpma, Dirk W.

    2012-01-01

    Tough networks are prepared by photo-crosslinking high-molecular-weight DLLA and TMC macromers. These amorphous networks exhibit tunable thermal and mechanical properties and have excellent shape-memory features. Variation of the monomer ratio allows adjustment of Tg between approximately −13 and

  1. Structure characterization of the central repetitive domain of high molecular weight gluten proteins .2. Characterization in solution and in the dry state

    NARCIS (Netherlands)

    van Dijk, A.A.; De Boef, E.; Bekkers, A.; van Wijk, L.L.; van Swieten, E.; Hamer, R.J.; Robillard, G.T.

    The structure of the central repetitive domain of high molecular weight (HMW) wheat gluten proteins was characterized in solution and in the dry state using HMW proteins Bx6 and Bx7 and a subcloned, bacterially expressed part of the repetitive domain of HMW Dx5. Model studies of the HMW consensus

  2. Structure characterization of the central repetitive domain of high molecular weight gluten proteins .1. Model studies using cyclic and linear peptides

    NARCIS (Netherlands)

    VanDijk, AA; VanWijk, LL; VanVliet, A; Haris, P; VanSwieten, E; Tesser, GI; Robillard, GT

    The high molecular weight (HMW) proteins from wheat contain a repetitive domain that forms 60-80% of their sequence. The consensus peptides PGQGQQ and GYYPTSPQQ form more than 90% of the domain; both are predicted to adopt beta-turn structure. This paper describes the structural characterization of

  3. Separation of mouse testis cells on a Celsep (TM) apparatus and their usefulness as a source of high molecular weight DNA or RNA

    Science.gov (United States)

    Wolgemuth, D. J.; Gizang-Ginsberg, E.; Engelmyer, E.; Gavin, B. J.; Ponzetto, C.

    1985-01-01

    The use of a self-contained unit-gravity cell separation apparatus for separation of populations of mouse testicular cells is described. The apparatus, a Celsep (TM), maximizes the unit area over which sedimentation occurs, reduces the amount of separation medium employed, and is quite reproducible. Cells thus isolated have been good sources for isolation of DNA, and notably, high molecular weight RNA.

  4. Enhancement of human adaptive immune responses by administration of a high-molecular-weight polysaccharide extract from the cyanobacterium Arthrospira platensis

    DEFF Research Database (Denmark)

    Pedersen, Morten Løbner; Walsted, Anette; Larsen, Rune

    2008-01-01

    The effect of consumption of Immulina, a high-molecular-weight polysaccharide extract from the cyanobacterium Arthrospira platensis, on adaptive immune responses was investigated by evaluation of changes in leukocyte responsiveness to two foreign recall antigens, Candida albicans (CA) and tetanus...

  5. The effect of chromic acid treatment on the mechanical and tribological properties of aramid fibre reinforced ultra-high molecular weight polyethylene composite

    NARCIS (Netherlands)

    Hofste, JM; Pennings, AJ; Schut, J.A.

    1998-01-01

    Surface oxidation of ultra-high molecular weight polyethylene (UHMWPE) powder has an influence on the mixing procedure of chopped fibres and UHMWPE powder. Due to this oxidation hydrogen bonds can be formed between the fibres and powder particles, leading to a more homogeneous fibre-powder mixture.

  6. In Vivo Biological Evaluation of High Molecular Weight Multifunctional Acid-Degradable Polymeric Drug Carriers with Structurally Different Ketals.

    Science.gov (United States)

    Shenoi, Rajesh A; Abbina, Srinivas; Kizhakkedathu, Jayachandran N

    2016-11-14

    Understanding the influence of degradable chemical moieties on in vivo degradation, tissue distribution, and excretion is critical for the design of novel biodegradable drug carriers. Polyketals have recently emerged as a promising therapeutic delivery platform due to their ability to degrade under mild acidic intracellular compartments and generation of nontoxic degradation products. However, the effect of chemical structure of the ketal groups on the in vivo degradation, biodistribution, and pharmacokinetics of water-soluble ketal-containing polymers has not been explored. In the present work, we synthesized high molecular weight, water-soluble biodegradable hyperbranched polyglycerols (BHPGs) through the incorporation of structurally different ketal groups into the main chain of highly biocompatible polyglycerols. BHPGs showed pH and ketal group structure dependent degradation in buffer solutions. When the polymers were intravenously administered in mice, a strong dependence of in vivo degradation, biodistribution, and clearance on the ketal group structure was observed. All the BHPGs demonstrated degradation and clearance in vivo, with minimal tissue accumulation. Interestingly, an unanticipated degradation behavior of BHPGs with structurally different ketal groups was observed in vivo in comparison to their degradation in buffer solutions. BHPGs with cyclohexyl ketal (CHK) and cyclopentyl ketal (CPK) groups degraded much faster and were cleared from circulation much rapidly, while BHPG with glycerol hydroxy butanone ketal (GHBK) group degraded at a much slower rate and exhibited similar plasma half-life as that of nondegradable HPG. BHPG-GHBK also showed significantly lower tissue accumulation than nondegradable HPG after 30 days of administration. The difference in in vivo degradation may be attributed to the difference in hydrophobic characteristics of different ketal containing polymers, which may change their interaction with proteins and cells in vivo

  7. Effects of ionizing radiation on the properties of ultra-high molecular weight polyethylene (PE-UHMW)

    International Nuclear Information System (INIS)

    Kurth, M.

    1990-01-01

    Ultra high molecular weight polyethylene (PE-UHMW) is used in most artificial joint replacement devices. Prior to implantation in biological environment, radiatin sterilization by 60 Co or electron beam is common. It is well known that polyethylene exposed to ionizing radiation of any sort undergo physical changes due to chain scission and/or crosslinking. PE-UHMW sheets, 8 mm thick, were either 60 Co or electron beam irradiated, in the range of 10-150 kGy under air or nitrogen atmoshere. The crystallinity of the irradiated samples increases with the irradiation dose. The chain scission/crosslinking events ratio determine the network structure and the sol/gel ratio. The latter was found to depend on irradiation dose, radiation atmosphere and sample thickness. Moreover 60 Co-irradiation is about 5 times more effective in forming PE-UHMW gel than electron-irradiation. Besides the degree of crosslinking, the molecular weight distribution is the main determinant of the structural properties of PE-UHMW. Low molecular weight fractions were also found. Using a dose of 30 kGy ( 60 Co in air), the average molecular weight of the soluble part after extraction decreased from originally 2.3 million g/mol to 170.000 g/mol, corresponding to a factor of about 10. These changes in molecular weight have a strong influence on the mechanical properties of PE-UHMW. Crosslinking slightly increases the yield strength, while the elongation at break decreases. Long-term compressive creep is reduced if the material is irradiated. Obviously, increased crystallinity after oxidative chain scission affects a higher deformation resistance. Radiation crosslinked structures cause a significant increase in abrasion resistance. The above described structural changes occur even upon irradiation of very low doses as used during sterilization. This study will enable to reduce the radiation sterilization damage and thus to gain long term stability of PE-UHMW medical devices. (orig./BBR)

  8. Ultra high molecular weight polyethylene (UHMWPE) fiber epoxy composite hybridized with Gadolinium and Boron nanoparticles for radiation shielding

    Science.gov (United States)

    Mani, Venkat; Prasad, Narasimha S.; Kelkar, Ajit

    2016-09-01

    Deep space radiations pose a major threat to the astronauts and their spacecraft during long duration space exploration missions. The two sources of radiation that are of concern are the galactic cosmic radiation (GCR) and the short lived secondary neutron radiations that are generated as a result of fragmentation that occurs when GCR strikes target nuclei in a spacecraft. Energy loss, during the interaction of GCR and the shielding material, increases with the charge to mass ratio of the shielding material. Hydrogen with no neutron in its nucleus has the highest charge to mass ratio and is the element which is the most effective shield against GCR. Some of the polymers because of their higher hydrogen content also serve as radiation shield materials. Ultra High Molecular Weight Polyethylene (UHMWPE) fibers, apart from possessing radiation shielding properties by the virtue of the high hydrogen content, are known for extraordinary properties. An effective radiation shielding material is the one that will offer protection from GCR and impede the secondary neutron radiations resulting from the fragmentation process. Neutrons, which result from fragmentation, do not respond to the Coulombic interaction that shield against GCR. To prevent the deleterious effects of secondary neutrons, targets such as Gadolinium are required. In this paper, the radiation shielding studies that were carried out on the fabricated sandwich panels by vacuum-assisted resin transfer molding (VARTM) process are presented. VARTM is a manufacturing process used for making large composite structures by infusing resin into base materials formed with woven fabric or fiber using vacuum pressure. Using the VARTM process, the hybridization of Epoxy/UHMWPE composites with Gadolinium nanoparticles, Boron, and Boron carbide nanoparticles in the form of sandwich panels were successfully carried out. The preliminary results from neutron radiation tests show that greater than 99% shielding performance was

  9. Effect of ZnO morphology on affecting bactericidal property of ultra high molecular weight polyethylene biocomposite

    International Nuclear Information System (INIS)

    Sharma, Rajeev Kumar; Agarwal, Meenakshi; Balani, Kantesh

    2016-01-01

    Bacterial infection of implants can be controlled by selective trapping of bacteria, followed with consequent killing by targeted antibacterial agents. Herein, the role of various ZnO morphologies, viz. micro-rods (R), nanoparticles (NP), and micro-disks (D) on antibacterial efficacy of ZnO via release of Zn"2"+ and H_2O_2 is assessed, both as isolated powders and via incorporating them in cytocompatible ultra high molecular weight polyethylene (UHMWPE). Though ZnO is antibacterial, interestingly, all ZnO morphologies elicited a supportive growth of gram-negative bacteria (Escherichia coli) in culture medium (until 28–35 μg/ml). But, all ZnO morphologies did elicit bactericidal effect on gram positive bacteria (Staphylococcus aureus or Staphylococcus epidermidis) both in culture medium (for 0–2.5 μg/ml) or when incorporated (5–20 wt.%) into UHMWPE. The bactericidal mechanisms were quantified for various ZnO morphologies via: (i) H_2O_2 production, (ii) Zn"2"+ release, and (iii) the presence of surface oxygen vacancies. On one hand, where only ZnO(NP) elicited release of H_2O_2 in the absence of light, maximum Zn"2"+ release was elicited by ZnO(D). Interestingly, when ZnO is incorporated as reinforcement (5–20 wt.%), its antibacterial action against E. coli was vividly observed due to selective proliferation of bacteria only on friendly UHMWPE matrix. Hence, luring bacteria on affable UHMWPE surface can be complemented with their targeted killing by ZnO present in composite. - Highlights: • The role of ZnO morphology in affecting bactericidal mechanisms • Quantification of Zn"2"+ release, H_2O_2 production and surface oxygen vacancy defects • Inherent resistance by gram negative bacteria at lower ZnO concentrations • Containment of bacteria on polymeric surface and consequent targeted killing by ZnO

  10. The interplay of plasma treatment and gold coating and ultra-high molecular weight polyethylene: On the cytocompatibility

    Energy Technology Data Exchange (ETDEWEB)

    Novotná, Zdenka, E-mail: zdenka1.novotn@vscht.cz [Department of Solid State Engineering, University of Chemistry and Technology Prague, Prague (Czech Republic); Rimpelová, Silvie; Juřík, Petr [Department of Biochemistry and Microbiology, University of Chemistry and Technology Prague, Prague (Czech Republic); Veselý, Martin [Department of Organic Technology, University of Chemistry and Technology Prague, Prague (Czech Republic); Kolská, Zdenka [Faculty and Science, J. E. Purkinje University in Usti nad Labem, Usti nad Labem (Czech Republic); Hubáček, Tomáš [Biology Centre CAS CR, SoWa National Research Infrastructure, Ceske Budejovice (Czech Republic); Ruml, Tomáš [Department of Biochemistry and Microbiology, University of Chemistry and Technology Prague, Prague (Czech Republic); Švorčík, Václav [Department of Solid State Engineering, University of Chemistry and Technology Prague, Prague (Czech Republic)

    2017-02-01

    We have investigated the application of Ar plasma for creation of nanostructured ultra high molecular weight polyethylene (PE) surface in order to enhance adhesion of mouse embryonic fibroblasts (L929). The aim of this study was to investigate the effect of the interface between plasma-treated and gold-coated PE on adhesion and spreading of cells. The surface properties of pristine samples and its modified counterparts were studied by different experimental techniques (gravimetry, goniometry and X-ray photoelectron spectroscopy (XPS), electrokinetic analysis), which were used for characterization of treated and sputtered layers, polarity and surface chemical structure, respectively. Further, atomic force microscopy (AFM) was employed to study the surface morphology and roughness. Biological responses of cells seeded on PE samples were evaluated in terms of cell adhesion, spreading, morphology and proliferation. Detailed cell morphology and intercellular connections were followed by scanning electron microscopy (SEM). As it was expected the thickness of a deposited gold film was an increasing function of the sputtering time. Despite the fact that plasma treatment proceeded in inert plasma, oxidized degradation products were formed on the PE surface which would contribute to increased hydrophilicity (wettability) of the plasma treated polymer. The XPS method showed a decrease in carbon concentration with increasing plasma treatment. Cell adhesion measured on the interface between plasma treated and gold coated PE was inversely proportional to the thickness of a gold layer on a sample. - Highlights: • Gold-coating improved wettability of polyethylene in comparison with plasma-treatment. • Plasma-treatment increased the surface roughness while the subsequent gold-coating decreased the roughness. • Adhesion and growth of mouse embryonic fibroblasts (L929) were studied in vitro. • Low amounts of gold nanoparticles released in the medium promoted cell growth.

  11. The interplay of plasma treatment and gold coating and ultra-high molecular weight polyethylene: On the cytocompatibility

    International Nuclear Information System (INIS)

    Novotná, Zdenka; Rimpelová, Silvie; Juřík, Petr; Veselý, Martin; Kolská, Zdenka; Hubáček, Tomáš; Ruml, Tomáš; Švorčík, Václav

    2017-01-01

    We have investigated the application of Ar plasma for creation of nanostructured ultra high molecular weight polyethylene (PE) surface in order to enhance adhesion of mouse embryonic fibroblasts (L929). The aim of this study was to investigate the effect of the interface between plasma-treated and gold-coated PE on adhesion and spreading of cells. The surface properties of pristine samples and its modified counterparts were studied by different experimental techniques (gravimetry, goniometry and X-ray photoelectron spectroscopy (XPS), electrokinetic analysis), which were used for characterization of treated and sputtered layers, polarity and surface chemical structure, respectively. Further, atomic force microscopy (AFM) was employed to study the surface morphology and roughness. Biological responses of cells seeded on PE samples were evaluated in terms of cell adhesion, spreading, morphology and proliferation. Detailed cell morphology and intercellular connections were followed by scanning electron microscopy (SEM). As it was expected the thickness of a deposited gold film was an increasing function of the sputtering time. Despite the fact that plasma treatment proceeded in inert plasma, oxidized degradation products were formed on the PE surface which would contribute to increased hydrophilicity (wettability) of the plasma treated polymer. The XPS method showed a decrease in carbon concentration with increasing plasma treatment. Cell adhesion measured on the interface between plasma treated and gold coated PE was inversely proportional to the thickness of a gold layer on a sample. - Highlights: • Gold-coating improved wettability of polyethylene in comparison with plasma-treatment. • Plasma-treatment increased the surface roughness while the subsequent gold-coating decreased the roughness. • Adhesion and growth of mouse embryonic fibroblasts (L929) were studied in vitro. • Low amounts of gold nanoparticles released in the medium promoted cell growth.

  12. A high molecular weight proteoglycan is differentially expressed during development of the mollusc Concholepas concholepas (Mollusca; Gastropoda; Muricidae).

    Science.gov (United States)

    Brandan, E; González, M; Inestrosa, N C; Tremblay, C; Urrea, R

    1992-12-15

    Incorporation of radioactive sulfate to hatched veliger larvae of the gastropod muricid Concholepas concholepas indicated that over 87% of the sulfated macromolecules were found in the detergent insoluble fraction, rich in extracellular matrix (ECM) components. The sulfated material was solubilized with guanidine salt followed by urea dialysis and fractionated by DEAE-Sephacel chromatography. Three sulfated compounds eluting at 0.7, 1.1, and 3.0 M NaCl, called peaks I, II, and III, respectively, were obtained. The sulfated compound present in peak I was degraded by pronase or sodium alkaline treatment to a small sulfated resistant material, suggesting the presence of a proteoglycan (PG). Filtration analysis on Sephacryl S-500 and SDS-PAGE of the intact PG indicates that it has a high molecular weight (360,000 to over 1 x 10(6)). Monoclonal antibodies (mAb) against this PG were produced. The specificity of one mAb, the 6H2, was demonstrated by size chromatography and ELISA analysis. The epitope recognized by this mAb seems to be present in the core protein of the PG. Both the extent of sulfation and the presence of different sulfated species of PGs were evaluated during the development of this mollusc. A twelvefold increase in the incorporation of sulfate to PGs per milligram of protein was found in veliger larvae compared to blastula-glastula stages. This change correlated well with the differential expression of the sulfated PG present in peak I. Biochemical and immunological analysis indicate that high levels of this PG are found in veliger and trocophore larvae in comparison with blastula-gastrula and early juveniles.(ABSTRACT TRUNCATED AT 250 WORDS)

  13. High molecular weight chitosan derivative polymeric micelles encapsulating superparamagnetic iron oxide for tumor-targeted magnetic resonance imaging

    Directory of Open Access Journals (Sweden)

    Xiao Y

    2015-02-01

    Full Text Available Yunbin Xiao,1,* Zuan Tao Lin,2,* Yanmei Chen,1 He Wang,1 Ya Li Deng,2 D Elizabeth Le,3 Jianguo Bin,1 Meiyu Li,1 Yulin Liao,1 Yili Liu,1 Gangbiao Jiang,2 Jianping Bin1 1State Key Laboratory of Organ Failure Research, Division of Cardiology, Nanfang Hospital, Southern Medical University, Guangzhou, People’s Republic of China; 2Department of Pharmaceutical Engineering, South China Agricultural University, Guangzhou, People’s Republic of China; 3Cardiovascular Division, Oregon Health and Science University, Portland, OR, USA *These authors contributed equally to this work Abstract: Magnetic resonance imaging (MRI contrast agents based on chitosan derivatives have great potential for diagnosing diseases. However, stable tumor-targeted MRI contrast agents using micelles prepared from high molecular weight chitosan derivatives are seldom reported. In this study, we developed a novel tumor-targeted MRI vehicle via superparamagnetic iron oxide nanoparticles (SPIONs encapsulated in self-aggregating polymeric folate-conjugated N-palmitoyl chitosan (FAPLCS micelles. The tumor-targeting ability of FAPLCS/SPIONs was demonstrated in vitro and in vivo. The results of dynamic light scattering experiments showed that the micelles had a relatively narrow size distribution (136.60±3.90 nm and excellent stability. FAPLCS/SPIONs showed low cytotoxicity and excellent biocompatibility in cellular toxicity tests. Both in vitro and in vivo studies demonstrated that FAPLCS/SPIONs bound specifically to folate receptor-positive HeLa cells, and that FAPLCS/SPIONs accumulated predominantly in established HeLa-derived tumors in mice. The signal intensities of T2-weighted images in established HeLa-derived tumors were reduced dramatically after intravenous micelle administration. Our study indicates that FAPLCS/SPION micelles can potentially serve as safe and effective MRI contrast agents for detecting tumors that overexpress folate receptors. Keywords: superparamagnetic

  14. FY 1991 Report on the results of the research and development of silicon-based high-molecular-weight materials; 1991 nendo keisokei kobunshi zairyo no kenkyu kaihatsu seika hokokusho

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1992-03-01

    The research and development project has been started to establish the basic technologies for molecular designs, synthesis, material production and evaluation of silicon-based high-molecular-weight materials expected to exhibit excellent characteristics, e.g., electro-optical functions, resistance to heat, flame retardance and mechanical properties. The efforts in FY 1991, the first year for the 10-year project, are mainly directed to the surveys on the R and D trends, both domestic and foreign, to clarify the relationship between the structures and functions/properties. The R and D projects followed include the technologies for synthesizing (1) electroconductive silicon-based high-molecular-weight materials, (2) novel silicon-based high-molecular-weight materials capable of drawing circuits, (3) novel, light-emitting silicon-based high-molecular-weight materials and (4) silicon-based opto-electric conversion materials for the electro-optical functional high-molecular-weight materials; and (1) synthesis of high-molecular-weight structural materials of sea island structure, (2) technologies for forming inter-penetrating type structures (IPN), (3) development of composite structural materials of organometallic complex and silicon-based high-molecular-weight material, and (4) development of silicon-based high-molecular-weight materials of ring structure for the high-molecular-weight structural materials. (NEDO)

  15. Effect of ZnO morphology on affecting bactericidal property of ultra high molecular weight polyethylene biocomposite

    Energy Technology Data Exchange (ETDEWEB)

    Sharma, Rajeev Kumar [Biomaterials Processing and Characterization Laboratory, Indian Institute of Technology Kanpur, Kanpur -208016 (India); Agarwal, Meenakshi [Amity Institute of Nanotechnology, Amity University, Noida, Uttar Pradesh - 201303 (India); Balani, Kantesh, E-mail: kbalani@iitk.ac.in [Biomaterials Processing and Characterization Laboratory, Indian Institute of Technology Kanpur, Kanpur -208016 (India)

    2016-05-01

    Bacterial infection of implants can be controlled by selective trapping of bacteria, followed with consequent killing by targeted antibacterial agents. Herein, the role of various ZnO morphologies, viz. micro-rods (R), nanoparticles (NP), and micro-disks (D) on antibacterial efficacy of ZnO via release of Zn{sup 2+} and H{sub 2}O{sub 2} is assessed, both as isolated powders and via incorporating them in cytocompatible ultra high molecular weight polyethylene (UHMWPE). Though ZnO is antibacterial, interestingly, all ZnO morphologies elicited a supportive growth of gram-negative bacteria (Escherichia coli) in culture medium (until 28–35 μg/ml). But, all ZnO morphologies did elicit bactericidal effect on gram positive bacteria (Staphylococcus aureus or Staphylococcus epidermidis) both in culture medium (for 0–2.5 μg/ml) or when incorporated (5–20 wt.%) into UHMWPE. The bactericidal mechanisms were quantified for various ZnO morphologies via: (i) H{sub 2}O{sub 2} production, (ii) Zn{sup 2+} release, and (iii) the presence of surface oxygen vacancies. On one hand, where only ZnO(NP) elicited release of H{sub 2}O{sub 2} in the absence of light, maximum Zn{sup 2+} release was elicited by ZnO(D). Interestingly, when ZnO is incorporated as reinforcement (5–20 wt.%), its antibacterial action against E. coli was vividly observed due to selective proliferation of bacteria only on friendly UHMWPE matrix. Hence, luring bacteria on affable UHMWPE surface can be complemented with their targeted killing by ZnO present in composite. - Highlights: • The role of ZnO morphology in affecting bactericidal mechanisms • Quantification of Zn{sup 2+} release, H{sub 2}O{sub 2} production and surface oxygen vacancy defects • Inherent resistance by gram negative bacteria at lower ZnO concentrations • Containment of bacteria on polymeric surface and consequent targeted killing by ZnO.

  16. Study of crosslinking onset and hydrogen annealing of ultra-high molecular weight polyethylene irradiated with high-energy protons

    Science.gov (United States)

    Wilson, John Ford

    1997-09-01

    Ultra high molecular weight polyethylene (UHMW-PE) is used extensively in hip and knee endoprostheses. Radiation damage from the sterilization of these endoprostheses prior to surgical insertion results in polymer crosslinking and decreased oxidative stability. The motivation for this study was to determine if UHMW-PE could be crosslinked by low dose proton irradiation with minimal radiation damage and its subsequent deleterious effects. I found that low dose proton irradiation and post irradiation hydrogen annealing did crosslink UHMW-PE and limit post irradiation oxidation. Crosslinking onset was investigated for UHMW-PE irradiated with 2.6 and 30 MeV H+ ions at low doses from 5.7 × 1011-2.3 × 1014 ions/cm2. Crosslinking was determined from gel permeation chromatography (GPC) of 1,2,4 trichlorobenzene sol fractions and increased with dose. Fourier transform infrared spectroscopy (FTIR) showed irradiation resulted in increased free radicals confirmed from increased carbonyl groups. Radiation damage, especially at the highest doses observed, also showed up in carbon double bonds and increased methyl end groups. Hydrogen annealing after ion irradiation resulted in 40- 50% decrease in FTIR absorption associated with carbonyl. The hydrogen annealing prevented further oxidation after aging for 1024 hours at 80oC. Hydrogen annealing was successful in healing radiation damage through reacting with the free radicals generated during proton irradiation. Polyethylenes, polyesters, and polyamides are used in diverse applications by the medical profession in the treatment of orthopedic impairments and cardiovascular disease and for neural implants. These artificial implants are sterilized with gamma irradiation prior to surgery and the resulting radiation damage can lead to accelerated deterioration of the implant properties. The findings in this study will greatly impact the continued use of these materials through the elimination of many problems associated with radiation

  17. High-molecular weight adiponectin/HOMA-IR ratio as a biomarker of metabolic syndrome in urban multiethnic Brazilian subjects.

    Science.gov (United States)

    de Abreu, Virgínia Genelhu; Martins, Cyro José de Moraes; de Oliveira, Patricia Aguiar Cardoso; Francischetti, Emilio Antonio

    2017-01-01

    Metabolic syndrome (MetS) has an important epidemiological relevance due to its increasing prevalence and association with type 2 diabetes and cardiovascular disease. Insulin resistance is a core feature of the MetS. HOMA-IR is a robust clinical and epidemiological marker of MetS. Adiponectin is an adipokine with insulin-sensitizing and anti-inflammatory functions; its levels decrease as number of components of MetS increases. High-molecular weight adiponectin (HMWA) is the multimer responsible for the relationship of adiponectin with insulin sensitivity. HOMA-IR and HMWA are suitable candidates for MetS biomarkers. The ratio of adiponectin to HOMA-IR has been validated as a powerful index of MetS and considered a better marker of its presence, than either HOMA-IR or adiponectin alone, in selected homogeneous populations. We compared the strength of association between HMWA, HOMA-IR and HMWA/HOMA-IR ratio with MetS and its key components. Our data have shown that the median (25th, 75th percentile) of HMWA/HOMA-IR ratio was lower in subjects with MetS [0.51 (0.33, 1.31)] as compared to those without it [2.19 (1.13, 4.71)]. The correlation coefficient (r) was significantly higher for HMWA/HOMA-IR ratio as compared to HMWA for waist circumference (-0.65; -0.40, respectively); mean blood pressure (-0.27; -0.14, respectively); fasting glucose (-0.38; -0.19, respectively); HDL-cholesterol (0.44; 0.40, respectively); and triglycerides (-0.35; -0.18, respectively). In a multivariable logistic regression analysis, the HMWA/HOMA-IR ratio was a sensitive predictor for MetS, being the only marker that was significantly associated with each and all the individual components of the syndrome. These results expand on previous studies in that we used the active circulating form of adiponectin, i.e. HMWA, and represent a typical Brazilian cohort characterized by intense interethnic admixture. Thus, the HMWA/HOMA-IR ratio is a minimally invasive biomarker for MetS that could be

  18. Inflammatory adipokines, high molecular weight adiponectin, and insulin resistance: a population-based survey in prepubertal schoolchildren.

    Directory of Open Access Journals (Sweden)

    Giuseppe Murdolo

    Full Text Available BACKGROUND: The aim of this study was to investigate sex differences and associations of high molecular weight (HMW adiponectin, leptin and proinflammatory adipokines, individually or in combinations, with adiposity and insulin resistance (IR measures in prepubertal childhood. METHODOLOGY: We studied 305 prepubertal children (boys/girls: 144/161; Tanner stage 1; age: 5-13 yr, included in a cohort of 44,231 adolescents who participated in an extensive Italian school-based survey. According to Cole's criteria, 105 individuals were lean (L; boys/girls: 59/46, 60 overweight (OW; boys/girls: 32/28 and 140 obese (OB; boys/girls: 70/70. Measurements comprised total and HMW adiponectin, leptin, as well as a panel of proinflammatory adipokines/chemokines associated with diabetes risk. PRINCIPAL FINDINGS: Leptin-, and the leptin-to-HMW adiponectin ratio (L/HMW-, increased progressively (p<0.0001 from L to OW to OB boys and girls. When compared with L peers, OW and OB girls exhibited lower (p<0.001 HMW adiponectin levels, while in boys the HMW multimers did not differ significantly across the BMI-stratified groups. OB girls displayed higher (p<0.05 IL-8, IL-18, monocyte chemoattractant protein-1 (MCP-1 and soluble intercellular adhesion molecule-1 levels (sICAM-1 than L girls, whereas increased macrophage migration inhibitory factor (MIF concentrations in OB vs OW boys were seen. HMW adiponectin (negatively, leptin or inflammatory markers (positively correlated with adiposity and IR measures. In multivariate models, leptin represented a strong and independent determinant of HOMA-IR (R(2 0.378; p<0.01. Adjustment for age, BMI(z-score, lipids and inflammatory mediators abolished the association between leptin and HOMA-IR in boys, while in girls leptin remained still a significant predictor of IR (R(2 0.513; p<0.01. Finally, in both sexes, the joint effect of the L/HMW did not improve the prediction of basal IR as compared with leptin levels alone, which were

  19. Selection of side-chain carbons in a high-molecular-weight, hydrophobic peptide using solid-state spectral editing methods

    International Nuclear Information System (INIS)

    Kumashiro, Kristin K.; Niemczura, Walter P.; Kim, Minna S.; Sandberg, Lawrence B.

    2000-01-01

    Solid-state spectral editing techniques have been used by others to simplify 13 C CPMAS spectra of small organic molecules, synthetic organic polymers, and coals. One approach utilizes experiments such as cross-polarization-with-polarization-inversion and cross-polarization-with-depolarization to generate subspectra. This work shows that this particular methodology is also applicable to natural-abundance 13 C CPMAS NMR studies of high-molecular-weight biopolymers. The editing experiments are demonstrated first with model peptides and then with α-elastin, a high-molecular-weight peptidyl preparation obtained from the elastic fibers in mammalian tissue. The latter has a predominance of small, nonpolar residues, which is evident in the crowded aliphatic region of typical 13 C CPMAS spectra. Spectral editing is particularly useful for simplifying the aliphatic region of the NMR spectrum of this elastin preparation

  20. Formation and stability of .beta.-structure in biodegradable ultra-high-molecular-weight poly(3-hydroxybutyrate) by infrared, Raman, and quantum chemical calculation studies

    Czech Academy of Sciences Publication Activity Database

    Murakami, R.; Sato, H.; Dybal, Jiří; Iwata, T.; Ozaki, Y.

    2007-01-01

    Roč. 48, č. 9 (2007), s. 2672-2680 ISSN 0032-3861 R&D Projects: GA ČR GA203/05/0425 Grant - others:Ministry of Education, Culture, Sports, Science and Technology(JP) 18750107 Institutional research plan: CEZ:AV0Z40500505 Keywords : ultra-high-molecular-weight poly(hydroxybutyrate) * .beta.-form * Raman spectroscopy Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.065, year: 2007

  1. High Molecular Weight Adiponectin Levels are Neither Influenced by Adiponectin Polymorphisms Nor Associated with Insulin Resistance in Mixed-Ancestry Hyperglycemic Subjects from South Africa

    Directory of Open Access Journals (Sweden)

    Zemlin Annalise E

    2016-10-01

    Full Text Available Background: High molecular weight (HMW adiponectin has antiatherogenic, antiinflammatory and antidiabetic properties and these effects have been linked to its effect on high density lipoprotein cholesterol (HDL-c. Single nucleotide polymorphisms (SNPs in the adiponectin gene influence adiponectin levels. We examined the relationship between HMW-adiponectin levels and cardiometabolic traits in normo- and hyperglycemic mixed ancestry South Africans and correlated these levels to two common polymorphisms.

  2. Characterization of low-molecular-weight glutenin subunit genes of ...

    Indian Academy of Sciences (India)

    2015-09-16

    Sep 16, 2015 ... 4Ministry of Education, Key Laboratory for Crop Genetic Resources and Improvement in Southwest China, ... gene mining have also been conducted in Aegilops genus, ... sites and pair-wise deletion for gaps/missing data.

  3. Characterization of low-molecular-weight glutenin subunit genes of ...

    Indian Academy of Sciences (India)

    Supplementary data, J. Genet. 94, 497–501. Table 2. LMW-GS genes obtained in this study. Species. Accession no. Clone. Length (bp). Sequence similarity. Ae. bicornis. CIae 47. Sb47-1. 1053. EU189089 96% T. aestivum. (SbSb). Sb47-2. 1053. EU189089 96% T. aestivum. Sb47-3. 1053. EU305550 97% Ae. longissima.

  4. Structure characterization of the central repetitive domain of high molecular weight gluten proteins. II. Characterization in solution and in the dry state

    OpenAIRE

    Dijk, Alard A. van; Boef, Esther de; Bekkers, August; Wijk, Lourens L. van; Swieten, Eric van; Hamer, Rob J.; Robillard, George T.

    1997-01-01

    The structure of the central repetitive domain of high molecular weight HMW) wheat gluten proteins was characterized in solution and in the dry state using HMW proteins Bx6 and Bx7 and a subcloned, bacterially expressed part of the repetitive domain of HMW Dx5. Model studies of the HMW consensus peptides PGQGQQ and GYYPTSPQQ formed the basis for the data analysis (van Dijk AA et al., 1997, Protein Sci 6:637-648). In solution, the repetitive domain contained a continuous nonoverlapping series ...

  5. High molecular weight non-polar hydrocarbons as pure model substances and in motor oil samples can be ionized without fragmentation by atmospheric pressure chemical ionization mass spectrometry.

    Science.gov (United States)

    Hourani, Nadim; Kuhnert, Nikolai

    2012-10-15

    High molecular weight non-polar hydrocarbons are still difficult to detect by mass spectrometry. Although several studies have targeted this problem, lack of good self-ionization has limited the ability of mass spectrometry to examine these hydrocarbons. Failure to control ion generation in the atmospheric pressure chemical ionization (APCI) source hampers the detection of intact stable gas-phase ions of non-polar hydrocarbon in mass spectrometry. Seventeen non-volatile non-polar hydrocarbons, reported to be difficult to ionize, were examined by an optimized APCI methodology using nitrogen as the reagent gas. All these analytes were successfully ionized as abundant and intact stable [M-H](+) ions without the use of any derivatization or adduct chemistry and without significant fragmentation. Application of the method to real-life hydrocarbon mixtures like light shredder waste and car motor oil was demonstrated. Despite numerous reports to the contrary, it is possible to ionize high molecular weight non-polar hydrocarbons by APCI, omitting the use of additives. This finding represents a significant step towards extending the applicability of mass spectrometry to non-polar hydrocarbon analyses in crude oil, petrochemical products, waste or food. Copyright © 2012 John Wiley & Sons, Ltd.

  6. Fabrication, Physicochemical Characterization, and Performance Evaluation of Biodegradable Polymeric Microneedle Patch System for Enhanced Transcutaneous Flux of High Molecular Weight Therapeutics.

    Science.gov (United States)

    Shah, Viral; Choudhury, Bijaya Krushna

    2017-11-01

    A revolutionary paradigm shift is being observed currently, towards the use of therapeutic biologics for disease management. The present research was focused on designing an efficient dosage form for transdermal delivery of α-choriogonadotropin (high molecular weight biologic), through biodegradable polymeric microneedles. Polyvinylpyrrolidone-based biodegradable microneedle arrays loaded with high molecular weight polypeptide, α-choriogonadotropin, were fabricated for its systemic delivery via transdermal route. Varied process and formulation parameters were optimized for fabricating microneedle array, which in turn was expected to temporally rupture the stratum corneum layer of the skin, acting as a major barrier to drug delivery through transdermal route. The developed polymeric microneedles were optimized on the basis of quality attributes like mechanical strength, axial strength, insertion ratio, and insertion force analysis. The optimized polymeric microneedle arrays were characterized for in vitro drug release studies, ex vivo drug permeation studies, skin resealing studies, and in vivo pharmacokinetic studies. Results depicted that fabricated polymeric microneedle arrays with mechanical strength of above 5 N and good insertion ratio exhibited similar systemic bioavailability of α-choriogonadotropin in comparison to marketed subcutaneous injection formulation of α-choriogonadotropin. Thus, it was ultimately concluded that the designed drug delivery system can serve as an efficient tool for systemic delivery of therapeutic biologics, with an added benefit of overcoming the limitations of parenteral delivery, achieving better patient acceptability and compliance.

  7. The relationship between seminal plasma zinc levels and high molecular weight zinc binding protein and sperm motility in Iraqi infertile men

    International Nuclear Information System (INIS)

    AbdulRasheed, Omar F

    2009-01-01

    To evaluate the relationship between sperm motility and total seminal plasma zinc concentration and high molecular weight zinc bound protein values in infertile Iraqi men. A case-control study was conducted at the Chemistry and Biochemistry Department, College of Medicine, Al-Nahrain University, Baghdad, Iraq between March 2005 to February 2006. The subjects for the study included 60 infertile male patients who were recruited Al-Kadhimiya Teaching Hospital, and Institute of Embryo Research and Infertility Treatment, Baghdad, Iraq. They were categorized according to their seminal parameters to oligozoospermia (n=32), azoospermia (n=22), and asthenozoospermia (n=6). Thirty nine fertile men (age range 31.87 +/- 3.76 years) were selected as controls, whose partners had conceived within the last year before participation with this study, and having normal spermiogram parameters. Seminal plasma zinc concentration and high molecular weight zinc binding proteins (HMW-Zn) were assayed in the ejaculates of fertile and infertile men. The seminal plasma zinc levels were 181.92 +/- 23.40 ug/mL in the oligozoospermia group, 178.50 +/- 18.61 ug/mL in the azoospermia group, 195.33 +/- 13.00 ug/mL in the asthenozoospermia group, and 184.66 +/- 21.31 ug/mL in the control group. The HMW-Zn% is a good index of sperm function rather than the total seminal plasma zinc levels. (author)

  8. Solid-State Polymerization of Poly(Ethylene Furanoate Biobased Polyester, II: An Efficient and Facile Method to Synthesize High Molecular Weight Polyester Appropriate for Food Packaging Applications

    Directory of Open Access Journals (Sweden)

    Nejib Kasmi

    2018-04-01

    Full Text Available The goal of this study was to synthesize, through a facile strategy, high molecular weight poly(ethylene furanoate (PEF, which could be applicable in food packaging applications. The efficient method to generate PEF with high molecular weight consists of carrying out a first solid-state polycondensation under vacuum for 6 h reaction time at 205 °C for the resulting polymer from two-step melt polycondensation process, which is catalyzed by tetrabutyl titanate (TBT. A remelting step was thereafter applied for 15 min at 250 °C for the obtained polyester. Thus, the PEF sample was ground into powder, and was then crystallized for 6 h at 170 °C. This polyester is then submitted to a second solid-state polycondensation (SSP carried out at different reaction times (1, 2, 3.5, and 5 h and temperatures 190, 200, and 205 °C, under vacuum. Ultimately, a significant increase in intrinsic viscosity is observed with only 5 h reaction time at 205 °C during the second SSP being needed to obtain very high molecular weight PEF polymer greater than 1 dL/g, which sufficient for manufacturing purposes. Intrinsic viscosity (IV, carboxyl end-group content (–COOH, and thermal properties, via differential scanning calorimetry (DSC, were measured for all resultant polyesters. Thanks to the post-polymerization process, DSC results showed that the melting temperatures of the prepared PEF samples were steadily enhanced in an obvious way as a function of reaction time and temperature increase. It was revealed, as was expected for all SSP samples, that the intrinsic viscosity and the average molecular weight of PEF polyester increased with increasing SSP time and temperature, whereas the number of carboxyl end-group concentration was decreased. A simple kinetic model was also developed and used to predict the time evolution of polyesters IV, as well as the carboxyl and hydroxyl end-groups of PEF during the SSP.

  9. The role of hydroperoxides as a precursor in the radiation-induced graft polymerization of methyl methacrylate to ultra-high molecular weight polyethylene

    Energy Technology Data Exchange (ETDEWEB)

    Enomoto, Ichiro, E-mail: enomoto.ichiro@iri-tokyo.j [Tokyo Metropolitan Industrial Technology Research Institute, KFC bldg., 12F, 1-6-1, Yokoami, Sumida-ku, Tokyo 130-0015 (Japan); School of Engineering, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8656 (Japan); Katsumura, Yosuke [School of Engineering, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8656 (Japan); Advanced Science Research Center, Japan Atomic Energy Agency, 2-4 Shirakata Shirane, Tokai-mura, Ibaraki 319-1195 (Japan); Kudo, Hisaaki [School of Engineering, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8656 (Japan); Sekiguchi, Masayuki [Tokyo Metropolitan Industrial Technology Research Institute, KFC bldg., 12F, 1-6-1, Yokoami, Sumida-ku, Tokyo 130-0015 (Japan)

    2010-06-15

    A graft polymerization of methyl methacrylate (MMA) to ultra-high molecular weight polyethylene (UHMWPE) with Co-60 gamma-ray irradiation in air at room temperature has been carried out. The grafting yields were measured as a function of the storage time (elapsed time from the end of irradiation to the start of grafting), and it was found that the yields reach at the maximum values at around several days since the end of irradiation. In order to clarify the precursor of the graft polymerization, changes of the radical yields and the carbonyl groups were measured as a function of storage time with ESR and microscopic FT-IR, respectively. From the similarities between the depth profiles of the hydroperoxide formation and the grafting products, it was concluded that the hydroperoxides can be main precursors of the grafting of the radiation-induced polymerization of MMA to UHMWPE under the given conditions.

  10. The role of hydroperoxides as a precursor in the radiation-induced graft polymerization of methyl methacrylate to ultra-high molecular weight polyethylene

    International Nuclear Information System (INIS)

    Enomoto, Ichiro; Katsumura, Yosuke; Kudo, Hisaaki; Sekiguchi, Masayuki

    2010-01-01

    A graft polymerization of methyl methacrylate (MMA) to ultra-high molecular weight polyethylene (UHMWPE) with Co-60 γ-ray irradiation in air at room temperature has been carried out. The grafting yields were measured as a function of the storage time (elapsed time from the end of irradiation to the start of grafting), and it was found that the yields reach at the maximum values at around several days since the end of irradiation. In order to clarify the precursor of the graft polymerization, changes of the radical yields and the carbonyl groups were measured as a function of storage time with ESR and microscopic FT-IR, respectively. From the similarities between the depth profiles of the hydroperoxide formation and the grafting products, it was concluded that the hydroperoxides can be main precursors of the grafting of the radiation-induced polymerization of MMA to UHMWPE under the given conditions.

  11. Physical properties of a high molecular weight hydroxyl-terminated polydimethylsiloxane modified castor oil based polyurethane/epoxy interpenetrating polymer network composites

    Science.gov (United States)

    Chen, Shoubing; Wang, Qihua; Wang, Tingmei

    2011-06-01

    A series of polyurethane (PU)/epoxy resin (EP) graft interpenetrating polymer network (IPN) composites modified by a high molecular weight hydroxyl-terminated polydimethylsiloxane (HTPDMS) were prepared. The effects of HTPDMS content on the phase structure, damping properties and the glass transition temperature ( Tg) of the HTPDMS-modified PU/EP IPN composites were studied by scanning electron microscopy (SEM) and dynamic mechanical analysis (DMA). Thermogravimetric analysis (TGA) showed that the thermal decomposition temperature of the composites increased with the increase of HTPDMS content. The tensile strength and impact strength of the IPN composites were also significantly improved, especially when the HTPDMS content was 10%. The modified IPN composites were expected to be used as structural damping materials in the future.

  12. Surfactant-Free RAFT Emulsion Polymerization of Styrene Using Thermoresponsive macroRAFT Agents: Towards Smart Well-Defined Block Copolymers with High Molecular Weights

    Directory of Open Access Journals (Sweden)

    Steffen Eggers

    2017-12-01

    Full Text Available The combination of reversible addition–fragmentation chain transfer (RAFT and emulsion polymerization has recently attracted much attention as a synthetic tool for high-molecular-weight block copolymers and their micellar nano-objects. Up to recently, though, the use of thermoresponsive polymers as both macroRAFT agents and latex stabilizers was impossible in aqueous media due to their hydrophobicity at the usually high polymerization temperatures. In this work, we present a straightforward surfactant-free RAFT emulsion polymerization to obtain thermoresponsive styrenic block copolymers with molecular weights of around 100 kDa and their well-defined latexes. The stability of the aqueous latexes is achieved by adding 20 vol % of the cosolvent 1,4-dioxane (DOX, increasing the phase transition temperature (PTT of the used thermoresponsive poly(N-acryloylpyrrolidine (PAPy macroRAFT agents above the polymerization temperature. Furthermore, this cosolvent approach is combined with the use of poly(N,N-dimethylacrylamide-block-poly(N-acryloylpiperidine-co-N-acryloylpyrrolidine (PDMA-b-P(APi-co-APy as the macroRAFT agent owning a short stabilizing PDMA end block and a widely adjustable PTT of the P(APi-co-APy block in between 4 and 47 °C. The temperature-induced collapse of the latter under emulsion polymerization conditions leads to the formation of RAFT nanoreactors, which allows for a very fast chain growth of the polystyrene (PS block. In dynamic light scattering (DLS, as well as cryo-transmission electron microscopy (cryoTEM, moreover, all created latexes indeed reveal a high (temperature stability and a reversible collapse of the thermoresponsive coronal block upon heating. Hence, this paper pioneers a versatile way towards amphiphilic thermoresponsive high-molecular-weight block copolymers and their nano-objects with tailored corona switchability.

  13. Preparation of High Purity, High Molecular-Weight Chitin from Ionic Liquids for Use as an Adsorbate for the Extraction of Uranium from Seawater

    Energy Technology Data Exchange (ETDEWEB)

    Rogers, Robin [Univ. of Alabama, Tuscaloosa, AL (United States)

    2013-12-21

    Ensuring a domestic supply of uranium is a key issue facing the wider implementation of nuclear power. Uranium is mostly mined in Kazakhstan, Australia, and Canada, and there are few high-grade uranium reserves left worldwide. Therefore, one of the most appealing potential sources of uranium is the vast quantity dissolved in the oceans (estimated to be 4.4 billion tons worldwide). There have been research efforts centered on finding a means to extract uranium from seawater for decades, but so far none have resulted in an economically viable product, due in part to the fact that the materials that have been successfully demonstrated to date are too costly (in terms of money and energy) to produce on the necessary scale. Ionic Liquids (salts which melt below 100{degrees}C) can completely dissolve raw crustacean shells, leading to recovery of a high purity, high molecular weight chitin powder and to fibers and films which can be spun directly from the extract solution suggesting that continuous processing might be feasible. The work proposed here will utilize the unprecedented control this makes possible over the chitin fiber a) to prepare electrospun nanofibers of very high surface area and in specific architectures, b) to modify the fiber surfaces chemically with selective extractant capacity, and c) to demonstrate their utility in the direct extraction and recovery of uranium from seawater. This approach will 1) provide direct extraction of chitin from shellfish waste thus saving energy over the current industrial process for obtaining chitin; 2) allow continuous processing of nanofibers for very high surface area fibers in an economical operation; 3) provide a unique high molecular weight chitin not available from the current industrial process, leading to stronger, more durable fibers; and 4) allow easy chemical modification of the large surface areas of the fibers for appending uranyl selective functionality providing selectivity and ease of stripping. The

  14. Preparation of High Purity, High Molecular-Weight Chitin from Ionic Liquids for Use as an Adsorbate for the Extraction of Uranium from Seawater

    International Nuclear Information System (INIS)

    Rogers, Robin

    2013-01-01

    Ensuring a domestic supply of uranium is a key issue facing the wider implementation of nuclear power. Uranium is mostly mined in Kazakhstan, Australia, and Canada, and there are few high-grade uranium reserves left worldwide. Therefore, one of the most appealing potential sources of uranium is the vast quantity dissolved in the oceans (estimated to be 4.4 billion tons worldwide). There have been research efforts centered on finding a means to extract uranium from seawater for decades, but so far none have resulted in an economically viable product, due in part to the fact that the materials that have been successfully demonstrated to date are too costly (in terms of money and energy) to produce on the necessary scale. Ionic Liquids (salts which melt below 100 deg C) can completely dissolve raw crustacean shells, leading to recovery of a high purity, high molecular weight chitin powder and to fibers and films which can be spun directly from the extract solution suggesting that continuous processing might be feasible. The work proposed here will utilize the unprecedented control this makes possible over the chitin fiber a) to prepare electrospun nanofibers of very high surface area and in specific architectures, b) to modify the fiber surfaces chemically with selective extractant capacity, and c) to demonstrate their utility in the direct extraction and recovery of uranium from seawater. This approach will 1) provide direct extraction of chitin from shellfish waste thus saving energy over the current industrial process for obtaining chitin; 2) allow continuous processing of nanofibers for very high surface area fibers in an economical operation; 3) provide a unique high molecular weight chitin not available from the current industrial process, leading to stronger, more durable fibers; and 4) allow easy chemical modification of the large surface areas of the fibers for appending uranyl selective functionality providing selectivity and ease of stripping. The resulting

  15. High molecular weight FGF2 isoforms demonstrate canonical receptor-mediated activity and support human embryonic stem cell self-renewal

    Directory of Open Access Journals (Sweden)

    Denis Kole

    2017-05-01

    Full Text Available Basic fibroblast growth factor (FGF2 is a highly pleiotropic member of a large family of growth factors with a broad range of activities, including mitogenesis and angiogenesis (Ornitz et al., 1996; Zhang et al., 2006, and it is known to be essential for maintenance of balance between survival, proliferation, and self-renewal in human pluripotent stem cells (Eiselleova et al., 2009; Zoumaro-Djayoon et al., 2011. A single FGF2 transcript can be translated into five FGF2 protein isoforms, an 18 kDa low molecular weight (LMW isoform and four larger high molecular weight (HMW isoforms (Arese et al., 1999; Arnaud et al., 1999. As they are not generally secreted, high molecular weight (HMW FGF2 isoforms have predominantly been investigated intracellularly; only a very limited number of studies have investigated their activity as extracellular factors. Here we report over-expression, isolation, and biological activity of all recombinant human FGF2 isoforms. We show that HMW FGF2 isoforms can support self-renewal of human embryonic stem cells (hESCs in vitro. Exogenous supplementation with HMW FGF2 isoforms also activates the canonical FGFR/MAPK pathway and induces mitogenic activity in a manner similar to that of the 18 kDa FGF2 isoform. Though all HMW isoforms, when supplemented exogenously, are able to recapitulate LMW FGF2 activity to some degree, it appears that certain isoforms tend to do so more poorly, demonstrating a lesser functional response by several measures. A better understanding of isoform-specific FGF2 effects will lead to a better understanding of developmental and pathological FGF2 signaling.

  16. RAFT Aqueous Dispersion Polymerization of N-(2-(Methacryloyloxy)ethyl)pyrrolidone: A Convenient Low Viscosity Route to High Molecular Weight Water-Soluble Copolymers.

    Science.gov (United States)

    Cunningham, Victoria J; Derry, Matthew J; Fielding, Lee A; Musa, Osama M; Armes, Steven P

    2016-06-28

    RAFT solution polymerization of N -(2-(methacryoyloxy)ethyl)pyrrolidone (NMEP) in ethanol at 70 °C was conducted to produce a series of PNMEP homopolymers with mean degrees of polymerization (DP) varying from 31 to 467. Turbidimetry was used to assess their inverse temperature solubility behavior in dilute aqueous solution, with an LCST of approximately 55 °C being observed in the high molecular weight limit. Then a poly(glycerol monomethacylate) (PGMA) macro-CTA with a mean DP of 63 was chain-extended with NMEP using a RAFT aqueous dispersion polymerization formulation at 70 °C. The target PNMEP DP was systematically varied from 100 up to 6000 to generate a series of PGMA 63 -PNMEP x diblock copolymers. High conversions (≥92%) could be achieved when targeting up to x = 5000. GPC analysis confirmed high blocking efficiencies and a linear evolution in M n with increasing PNMEP DP. A gradual increase in M w / M n was also observed when targeting higher DPs. However, this problem could be minimized ( M w / M n RAFT aqueous dispersion polymerization of NMEP was approximately four times faster than the RAFT solution polymerization of NMEP in ethanol when targeting the same DP in each case. This is perhaps surprising because both 1 H NMR and SAXS studies indicate that the core-forming PNMEP chains remain relatively solvated at 70 °C in the latter formulation. Moreover, dissolution of the initial PGMA 63 -PNMEP x particles occurs on cooling from 70 to 20 °C as the PNMEP block passes through its LCST. Hence this RAFT aqueous dispersion polymerization formulation offers an efficient route to a high molecular weight water-soluble polymer in a rather convenient low-viscosity form. Finally, the relatively expensive PGMA macro-CTA was replaced with a poly(methacrylic acid) (PMAA) macro-CTA. High conversions were also achieved for PMAA 85 -PNMEP x diblock copolymers prepared via RAFT aqueous dispersion polymerization for x ≤ 4000. Again, better control was achieved when

  17. 13CHD2–CEST NMR spectroscopy provides an avenue for studies of conformational exchange in high molecular weight proteins

    International Nuclear Information System (INIS)

    Rennella, Enrico; Huang, Rui; Velyvis, Algirdas; Kay, Lewis E.

    2015-01-01

    An NMR experiment for quantifying slow (millisecond) time-scale exchange processes involving the interconversion between visible ground state and invisible, conformationally excited state conformers is presented. The approach exploits chemical exchange saturation transfer (CEST) and makes use of 13 CHD 2 methyl group probes that can be readily incorporated into otherwise highly deuterated proteins. The methodology is validated with an application to a G48A Fyn SH3 domain that exchanges between a folded conformation and a sparsely populated and transiently formed unfolded ensemble. Experiments on a number of different protein systems, including a 360 kDa half-proteasome, establish that the sensitivity of this 13 CHD 2 13 C–CEST technique can be upwards of a factor of 5 times higher than for a previously published 13 CH 3 13 C–CEST approach (Bouvignies and Kay in J Biomol NMR 53:303–310, 2012), suggesting that the methodology will be powerful for studies of conformational exchange in high molecular weight proteins

  18. The Influence of Irradiation and Accelerated Aging on the Mechanical and Tribological Properties of the Graphene Oxide/Ultra-High-Molecular-Weight Polyethylene Nanocomposites

    Directory of Open Access Journals (Sweden)

    Guodong Huang

    2016-01-01

    Full Text Available Graphene oxide/ultra-high-molecular-weight polyethylene (GO/UHMWPE nanocomposite is a potential and promising candidate for artificial joint applications. However, after irradiation and accelerated aging, the mechanical and tribological behaviors of the nanocomposites are still unclear and require further investigation. GO/UHMWPE nanocomposites were successfully fabricated using ultrasonication dispersion, ball-milling, and hot-pressing process. Then, the nanocomposites were irradiated by gamma ray at doses of 100 kGy. Finally, GO/UHMWPE nanocomposites underwent accelerated aging at 80°C for 21 days in air. The mechanical and tribological properties of GO/UHMWPE nanocomposites have been evaluated after irradiation and accelerated aging. The results indicated that the incorporation of GO could enhance the mechanical, wear, and antiscratch properties of UHMWPE. After irradiation, these properties could be further enhanced, compared to unirradiated ones. After accelerated aging, however, these properties have been significantly reduced when compared to unirradiated ones. Moreover, GO and irradiation can synergistically enhance these properties.

  19. Effects of strain rate and temperature on the mechanical behavior of carbon black reinforced elastomers based on butyl rubber and high molecular weight polyethylene

    Science.gov (United States)

    Hussein, M.

    2018-06-01

    The influence of the mechanical property and morphology of different blend ratio of Butyl rubber (IIR)/high molecular weight polyethylene (PE) by temperature and strain rate are performed. Special attention has been considered to a ductile-brittle transition that is known to occur at around 60 °C. The idea is to explain the unexpected phenomenon of brittleness which directly related to all tensile mechanical properties such as the strength of blends, modulus of elasticity of filled and unfilled IIR-polyethylene blends. In particular, the initial Young's modulus, tensile strength and strain at failure exhibit similar dependency on strain rate and temperature. These quantities lowered and increased with an increment of temperature, whereas the increased with increasing of strain rate. Furthermore, the tensile strength and strain at failure decreases for all temperatures range with the increase of PE content in the blend, except Young's modulus in reverse. The strain rate sensitivity index parameter of the examined polymeric materials is consistent with the micro-mechanisms of deformation and the behavior was well described by an Eyring relationship leading to an activation volume of ∼1 nm3, except for the highest value of unfilled IIR ∼8.45 nm3.

  20. Improved Yield of High Molecular Weight DNA Coincides with Increased Microbial Diversity Access from Iron Oxide Cemented Sub-Surface Clay Environments

    Science.gov (United States)

    Hurt, Richard A.; Robeson, Michael S.; Shakya, Migun; Moberly, James G.; Vishnivetskaya, Tatiana A.; Gu, Baohua; Elias, Dwayne A.

    2014-01-01

    Despite over three decades of progress, extraction of high molecular weight (HMW) DNA from high clay soils or iron oxide cemented clay has remained challenging. HMW DNA is desirable for next generation sequencing as it yields the most comprehensive coverage. Several DNA extraction procedures were compared from samples that exhibit strong nucleic acid adsorption. pH manipulation or use of alternative ion solutions offered no improvement in nucleic acid recovery. Lysis by liquid N2 grinding in concentrated guanidine followed by concentrated sodium phosphate extraction supported HMW DNA recovery from clays high in iron oxides. DNA recovered using 1 M sodium phosphate buffer (PB) as a competitive desorptive wash was 15.22±2.33 µg DNA/g clay, with most DNA consisting of >20 Kb fragments, compared to 2.46±0.25 µg DNA/g clay with the Powerlyzer system (MoBio). Increasing PB concentration in the lysis reagent coincided with increasing DNA fragment length during initial extraction. Rarefaction plots of 16S rRNA (V1–V3 region) pyrosequencing from A-horizon and clay soils showed an ∼80% and ∼400% larger accessed diversity compared to the Powerlyzer soil DNA system, respectively. The observed diversity from the Firmicutes showed the strongest increase with >3-fold more operational taxonomic units (OTU) recovered. PMID:25033199

  1. Corneal protection with high-molecular-weight hyaluronan against in vitro and in vivo sodium lauryl sulfate-induced toxic effects.

    Science.gov (United States)

    Pauloin, Thierry; Dutot, Mélody; Liang, Hong; Chavinier, Emilie; Warnet, Jean-Michel; Rat, Patrice

    2009-10-01

    The aim of this study was to investigate high-molecular-weight hyaluronan (HA-HMW) corneal protection against sodium lauryl sulfate (SLS)-induced toxic effects with in vitro and in vivo experimental approaches. In vitro experiments consisted of a human corneal epithelial cell line incubated with HA-HMW, rinsed, and incubated with SLS. Cell viability, oxidative stress, chromatin condensation, caspase-3, -8, -9, and P2X7 cell death receptor activation, interleukin-6, and interleukin-8 production were investigated. In vivo experiments consisted of 36 New Zealand white rabbits treated for 3 days, 3 times per day, with HA-HMW or phosphate-buffered salt solution. At day 4, eyes were treated with SLS. Clinical observation and in vivo confocal microscopy using the Rostock Cornea Module of the Heidelberg Retina Tomograph-II were performed to evaluate and to compare SLS-induced toxicity between eyes treated with HA-HMW and eyes treated with phosphate-buffered salt solution. In vitro data indicate that exposure of human corneal epithelial cells to HA-HMW significantly decreased SLS-induced oxidative stress, apoptosis, and inflammation cytokine production. In vivo data indicate that SLS cornea injuries, characterized by damaged corneal epithelium, damaged anterior stroma, and inflammatory infiltrations, were attenuated with HA-HMW treatment. A good correlation was seen between in vitro and in vivo findings showing that HA-HMW decreases SLS-induced toxic effects and protects cornea.

  2. Gradient Architecture as Means of Phase Diagram Manipulation in Copolymers: Accessing Both LCOT and UCOT in High Molecular Weight Styrene/n-Butyl Acrylate Systems

    Science.gov (United States)

    Mok, Michelle; Burghardt, Wesley; Ellison, Christopher; Torkelson, John

    2009-03-01

    Traditionally, phase transitions of block copolymers could only be tuned through molecular weight and relative block length. Here, we introduce comonomer sequence design through gradient compositions as a means of further manipulating phase diagram boundaries. In such gradient copolymers, the reduced repulsion between chain segments allows access to phase transitions even at high molecular weights (MW). Rheological and x-ray scattering studies were performed to study the impact of comonomer sequence on phase behavior in styrene/n-butyl acrylate (S/nBA) systems. In S/nBA block copolymers, only upper critical ordering behavior was observed. In contrast, by using a gradient architecture of higher MW we observed both upper and lower ordering transitions similar to those seen in very weakly segregating S/n-butyl methacrylate block copolymers, where such dual ordering transitions were first detected by Russell et al. This is the first study to access a miscibility gap in gradient copolymers. Access to such behavior is very rare in blends and block copolymers, limited to low MW and/or very weakly segregating systems.

  3. Electrochemical immunoassay for the prostate specific antigen using a reduced graphene oxide functionalized with a high molecular-weight silk peptide

    International Nuclear Information System (INIS)

    Wang, Yanying; Qu, Ying; Li, Chunya; Wu, Kangbing; Liu, Guishen; Hou, Xiaodong; Huang, Yina; Wu, Wangze

    2015-01-01

    High molecular-weight silk peptide (SP) was used to functionalize the surface of nanosheets of reduced graphene oxide (rGO). The SP-rGO nanocomposite was then mixed with mouse anti-human prostate specific antigen monoclonal antibody (anti-PSA) and coated onto a glassy carbon electrode to fabricate an immunosensor. By using the hexacyanoferrate redox system as electroactive probe, the immunosensor was characterized by voltammetry and electrochemical impedance spectroscopy. The peak current, measured at the potential of 0.24 V (vs. SCE), is distinctly reduced after binding prostate specific antigen (PSA). Response (measured by differential pulse voltammetry) is linearly related to PSA concentration in the range from 0.1 to 5.0 ng · mL −1 and from 5.0 to 80.0 ng∙mL −1 , and the detection limit is 53 pg∙mL −1 (at an SNR of 3). The immunosensor was successfully applied to the determination of PSA in clinical serum samples, and the results were found to agree well with those obtained with an enzyme-linked immunosorbent assay. (author)

  4. Wear of ultra-high molecular weight polyethylene against damaged and undamaged stainless steel and diamond-like carbon-coated counterfaces.

    Science.gov (United States)

    Firkins, P; Hailey, J L; Fisher, J; Lettington, A H; Butter, R

    1998-10-01

    The wear of ultra-high molecular weight polyethylene (UHMWPE) in artificial joints and the resulting wear debris-induced osteolysis remains a major clinical concern in the orthopaedic sector. Third-body damage of metallic femoral heads is often cited as a cause of accelerated polyethylene wear, and the use of ceramic femoral heads in the hip is gaining increasing favour. In the knee prostheses and for smaller diameter femoral heads, the application of hard surface coatings, such as diamond-like carbon, is receiving considerable attention. However, to date, there has been little or no investigation of the tribology of these coatings in simulated biological environments. In this study, diamond-like carbon (DLC) has been compared to stainless steel in its undamaged form and following simulated third-body damage. The wear of UHMWPE was found to be similar when sliding against undamaged DLC and stainless steel counterfaces. DLC was found to be much more damage resistant than DLC. Under test conditions that simulate third-body damage to the femoral head, the wear of UHMWPE was seven times lower against DLC than against stainless steel (P < 0.05). The study shows DLC has considerable potential as a femoral bearing surface in artificial joints.

  5. Leukocyte Telomere Length and Serum Levels of High-Molecular-Weight Adiponectin and Dehydroepiandrosterone-Sulfate Could Reflect Distinct Aspects of Longevity in Japanese Centenarians

    Directory of Open Access Journals (Sweden)

    Yuji Aoki MD, PhD

    2017-03-01

    Full Text Available Leukocyte telomere length and serum levels of high-molecular-weight adiponectin and dehydroepiandrosterone-sulfate (DHEA-S were assessed in association with nutrition and performance status (PS in Japanese centenarians. Twenty-three centenarians (five men, 18 women were classified according to their PS 1 (nearly fully ambulatory, n = 2, 2 (in bed less than 50% of daytime, n = 10, 3 (in bed greater than 50%, n = 6, and 4 (completely bedridden, n = 5. Leukocyte telomere length was determined by the hybridization protection assay, and the adiponectin and DHEA-S levels were measured by chemiluminescent enzyme immunoassay. Among variables of PS, body mass index (BMI, albumin, adiponectin, DHEA-S, and telomere length, there were significant correlations between PS and albumin ( r = −.694, p < .01, between telomere length and BMI ( r = .522, p < .05, between adiponectin and BMI ( r = −.574, p < .01, and between DHEA-S and albumin ( r = .530, p < .01. When excluding two cancer-bearing centenarians with short telomere, telomere length significantly correlated with PS ( r = −.632, p < .01. It was indicated that the short leukocyte telomere was associated with poor PS and cancer development and that the adiponectin or DHEA-S was associated with adiposity or nutritional status. Despite a small number of subjects, these biomarkers seemed to reflect distinct aspects of longevity in Japanese centenarians.

  6. Leukocyte Telomere Length and Serum Levels of High-Molecular-Weight Adiponectin and Dehydroepiandrosterone-Sulfate Could Reflect Distinct Aspects of Longevity in Japanese Centenarians.

    Science.gov (United States)

    Aoki, Yuji; Aoki, Masato; Yamada, Kazuya

    2017-01-01

    Leukocyte telomere length and serum levels of high-molecular-weight adiponectin and dehydroepiandrosterone-sulfate (DHEA-S) were assessed in association with nutrition and performance status (PS) in Japanese centenarians. Twenty-three centenarians (five men, 18 women) were classified according to their PS 1 (nearly fully ambulatory, n = 2), 2 (in bed less than 50% of daytime, n = 10), 3 (in bed greater than 50%, n = 6), and 4 (completely bedridden, n = 5). Leukocyte telomere length was determined by the hybridization protection assay, and the adiponectin and DHEA-S levels were measured by chemiluminescent enzyme immunoassay. Among variables of PS, body mass index (BMI), albumin, adiponectin, DHEA-S, and telomere length, there were significant correlations between PS and albumin ( r = -.694, p r = .522, p r = -.574, p r = .530, p r = -.632, p < .01). It was indicated that the short leukocyte telomere was associated with poor PS and cancer development and that the adiponectin or DHEA-S was associated with adiposity or nutritional status. Despite a small number of subjects, these biomarkers seemed to reflect distinct aspects of longevity in Japanese centenarians.

  7. Enzymatic and acidic degradation of high molecular weight dextran into low molecular weight and its characterizations using novel Diffusion-ordered NMR spectroscopy.

    Science.gov (United States)

    Iqbal, Samina; Marchetti, Roberta; Aman, Afsheen; Silipo, Alba; Qader, Shah Ali Ul; Molinaro, Antonio

    2017-10-01

    Low molecular weight fractions were derived from native high molecular weight dextran produced by Leuconostoc mesenteroides KIBGE-IB26. Structural characterization of native and low molecular weight fractions obtained after acidic and enzymatic hydrolysis was done using FTIR and NMR spectroscopy. The molecular weight was estimated using Diffusion Ordered NMR spectroscopy. Native dextran (892kDa) is composed of α-(1→6) glycosidic linkage along with α-(1→3) branching. Major proportion of 528kDa dextran was obtained after prolong enzymatic hydrolysis however, an effective acidic treatment at pH-1.4 up to 02 and 04h of exposure resulted in the formation of 77kDa and 57kDa, respectively. The increment in pH from 1.4 to 1.8 lowered the hydrolysis efficiency and resulted in the formation of 270kDa dextran fraction. The results suggest that derived low molecular weight water soluble fractions can be utilized as a drug delivery carrier along with multiple application relating pharmaceutical industries. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Radiation grafting of various water-soluble monomers on ultra-high molecular weight polyethylene powder. Part II: Thermal, FTIR and morphological characterisation

    Energy Technology Data Exchange (ETDEWEB)

    Aydinli, Bahattin; Tincer, Teoman E-mail: teotin@metu.edu.tr

    2001-10-01

    Radiation induced grafted polyacrylic acid (PAA), polymethacrylic acid (PMAA), polyacrylamide (PAAm), poly N,N-dimethyl acrylamide (PNDAAm) and poly 1-vinyl-2 pyrrolidone (PVP) on ultra-high molecular weight polyethylene (UHMWPE) were characterised by DSC, FTIR and SEM analysis. While the effect of irradiation on pure UHMWPE was found to increase crystallinity and cause higher enthalpy of crystallisation, grafted UHMWPE powders showed lower crystallinity and enthalpy of crystallisation. In all grafted UHMWPE there existed secondary transitions corresponding to grafting polymers in the first run of DSC above 60 deg. C and they became clearer at a higher grafting level. In the second run of DSC some T{sub g} values appeared to shift to higher temperatures while some were not detected. FTIR analysis indicated the presence of water-soluble polymers in the grafted UHMWPE. The characteristic peaks of water-soluble polymers became sharper in the grafted UHMWPE. SEM analysis revealed that the grafting occurs both on fiber and microparticles of UHMWPE while flowing characteristic of powder is retained.

  9. Radiation grafting of various water-soluble monomers on ultra-high molecular weight polyethylene powder. Part II: Thermal, FTIR and morphological characterisation

    Science.gov (United States)

    Aydınlı, Bahattin; Tin c̡er, Teoman

    2001-10-01

    Radiation induced grafted polyacrylic acid (PAA), polymethacrylic acid (PMAA), polyacrylamide (PAAm), poly N,N-dimethyl acrylamide (PNDAAm) and poly 1-vinyl-2 pyrrolidone (PVP) on ultra-high molecular weight polyethylene (UHMWPE) were characterised by DSC, FTIR and SEM analysis. While the effect of irradiation on pure UHMWPE was found to increase crystallinity and cause higher enthalpy of crystallisation, grafted UHMWPE powders showed lower crystallinity and enthalpy of crystallisation. In all grafted UHMWPE there existed secondary transitions corresponding to grafting polymers in the first run of DSC above 60°C and they became clearer at a higher grafting level. In the second run of DSC some Tg values appeared to shift to higher temperatures while some were not detected. FTIR analysis indicated the presence of water-soluble polymers in the grafted UHMWPE. The characteristic peaks of water-soluble polymers became sharper in the grafted UHMWPE. SEM analysis revealed that the grafting occurs both on fiber and microparticles of UHMWPE while flowing characteristic of powder is retained.

  10. Mechanochemical Ring-Opening Polymerization of Lactide: Liquid-Assisted Grinding for the Green Synthesis of Poly(lactic acid) with High Molecular Weight.

    Science.gov (United States)

    Ohn, Nuri; Shin, Jihoon; Kim, Sung Sik; Kim, Jeung Gon

    2017-09-22

    Mechanochemical polymerization of lactide is carried out by using ball milling. Mechanical energy from collisions between the balls and the vessel efficiently promotes an organic-base-mediated metal- and solvent-free solid-state polymerization. Investigation of the parameters of the ball-milling synthesis revealed that the degree of lactide ring-opening polymerization could be modulated by the ball-milling time, vibration frequency, mass of the ball media, and liquid-assisted grinding. Liquid-assisted grinding was found to be an especially important factor for achieving a high degree of mechanochemical polymerization. Although polymer-chain scission from the strong collision energy prevented mechanical-force-driven high-molecular-weight polymer synthesis, the addition of only a small amount of liquid enabled sufficient energy dissipation and poly(lactic acid) was thereby obtained with a molecular weight of over 1×10 5  g mol -1 . © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Sustained High Levels of Both Total and High Molecular Weight Adiponectin in Plasma during the Convalescent Phase of Haemorrhagic Fever with Renal Syndrome Are Associated with Disease Severity

    Directory of Open Access Journals (Sweden)

    Kang Tang

    2017-01-01

    Full Text Available Haemorrhagic fever with renal syndrome (HFRS is characterised by an uncontrolled immune response that causes vascular leakage. Adiponectin (APN is an adipocytokine involved in prorevascularisation and immunomodulation. To investigate the possible effects of APN in the pathogenesis of HFRS, total and high molecular weight (HMW APN levels in the plasma of patients with HFRS were quantified using enzyme-linked immunosorbent assay (ELISA. Compared with those in healthy controls, the plasma total and HMW APN levels in patients were elevated to different degrees from the fever onset and remained high at the convalescent phase. Consistent with these results, western blot analysis additionally showed that low molecular weight (LMW, middle molecular weight (MMW, and HMW APN levels were all elevated and contributed to the elevation of the total APN level. Importantly, sustained high levels of total and HMW APN at the convalescent phase were significantly higher in patients with critical disease than those in patients with mild or moderate disease. Moreover, total and HMW APN levels negatively correlated with white blood cell count and positively correlated with platelet count and serum albumin level. These results may provide insights into understanding the roles of total and HMW APN in the pathogenesis of HFRS.

  12. Characterization of the mechanical properties of a new grade of ultra high molecular weight polyethylene and modeling with the viscoplasticity based on overstress.

    Science.gov (United States)

    Khan, Fazeel; Yeakle, Colin; Gomaa, Said

    2012-02-01

    Enhancements to the service life and performance of orthopedic implants used in total knee and hip replacement procedures can be achieved through optimization of design and the development of superior biocompatible polymeric materials. The introduction of a new or modified polymer must, naturally, be preceded by a rigorous testing program. This paper presents the assessment of the mechanical properties of a new filled grade of ultra high molecular weight polyethylene (UHMWPE) designated AOX(TM) and developed by DePuy Orthopaedics Inc. The deformation behavior was investigated through a series of tensile and compressive tests including strain rate sensitivity, creep, relaxation, and recovery. The polymer was found to exhibit rate-reversal behavior for certain loading histories: strain rate during creep with a compressive stress can be negative, positive, or change between the two during a test. Analogous behavior occurs during relaxation as well. This behavior lies beyond the realm of most numerical models used to computationally investigate and improve part geometry through finite element analysis of components. To address this shortcoming, the viscoplasticity theory based on overstress (VBO) has been suitably modified to capture these trends. VBO is a state variable based model in a differential formulation. Numerical simulation and prediction of all of the aforementioned tests, including good reproduction of the rate reversal behavior, is presented in this study. Copyright © 2011 Elsevier Ltd. All rights reserved.

  13. Influence of the graphite type in the mechanical properties of graphene nanosheets and ultra high molecular weight polyethylene (GN/UHMWPE) nanocomposites

    International Nuclear Information System (INIS)

    Lima, M.E. de A.; Coutinho, S.V.; Fim, F. de C.

    2016-01-01

    This paper studies the mechanical properties of graphene nanosheets and ultra high molecular weight polyethylene (GN/UHMWPE) nanocomposites. The graphene nanosheets were obtained by two methods: the first, from Graphite Micrograf HC11 after 8 hours in a ultrasound bath in 70% alcoholic solution; the second, from Graflake 9950 prepared by oxidizing with H 2 SO 4 /HNO 3 (4:1), followed by thermal expansion at 1050°C for 30 seconds and ultrasound bath for 20 hours. Both were oven dried for 24 hours at 110°C and subsequently added to the polymer in percentages of 0.5, 1.0 and 1.5% (w/w). The powders of the nanocomposites were mixed in a ball mill for 1 hour and molded by hot compression. XRD and SEM images was possible to verify that the Graflake's GN had higher delamination that the Micrograph's GN. The mechanical properties were affected in different percentages for each graphite type. (author)

  14. Marker-Assisted Selection for Recognizing Wheat Mutant Genotypes Carrying HMW Glutenin Alleles Related to Baking Quality

    OpenAIRE

    Zamani, Mohammad Javad; Bihamta, Mohammad Reza; Naserian Khiabani, Behnam; Tahernezhad, Zahra; Hallajian, Mohammad Taher; Shamsi, Marzieh Varasteh

    2014-01-01

    Allelic diversity of HMW glutenin loci in several studies revealed that allelic combinations affect dough quality. Dx5 + Dy10 subunits are related to good baking quality and Dx2 + Dy12 are related to undesirable baking quality. One of the most regular methods to evaluate the baking quality is SDS-PAGE which is used to improve baking quality labs. Marker-assisted selection is the method which can recognize the alleles related to baking quality and this method is based on polymerase chain reac...

  15. Potential impact of renin-angiotensin system inhibitors and calcium channel blockers on plasma high-molecular-weight adiponectin levels in hemodialysis patients

    International Nuclear Information System (INIS)

    Nakagawa, Naoki; Yao, Naoyuki; Hirayama, Tomoya

    2011-01-01

    Although metabolic syndrome confers an increased risk of cardiovascular disease in the general population, little is known about the alteration of abdominal adiposity and its association with adipocytokines in hemodialysis patients. We investigated the plasma high-molecular-weight (HMW) adiponectin level and its relationship to visceral fat area (VFA) and various markers of atherosclerosis in hemodialysis patients. In a cross-sectional study, conventional cardiovascular risk factors, plasma total and HMW adiponectin, the number of components of the metabolic syndrome and, using computed tomography, the distribution of abdominal adiposity were assessed in 144 hemodialysis patients (90 men and 54 women; mean age, 60.7 years) and 30 age- and sex-matched patients with chronic kidney disease (CKD). Plasma HMW adiponectin levels in hemodialysis patients were significantly higher than those in patients with CKD, negatively associated with VFA and serum triglycerides and positively associated with plasma total adiponectin, as well as the HMW-to-total adiponectin ratio in men and women (all P<0.05) in a simple regression analysis. In a multiple regression analysis, VFA was a significant determinant of HMW adiponectin in hemodialysis patients. Furthermore, after adjustment for classical risk factors, HMW adiponectin levels were significantly higher in patients undergoing treatment with renin-angiotensin system inhibitors or calcium channel blockers compared with patients not undergoing such treatment. This study shows that plasma HMW adiponectin levels were negatively associated with VFA and positively associated with treatment with blockade of the renin-angiotensin system and of the calcium channel. Therefore, these drugs might be effective for improving adipocytokine-related metabolic abnormalities in hemodialysis patients. (author)

  16. High-molecular-weight adiponectin is selectively reduced in women with polycystic ovary syndrome independent of body mass index and severity of insulin resistance.

    LENUS (Irish Health Repository)

    O'Connor, A

    2010-03-01

    Context: High-molecular-weight (HMW) adiponectin contributes to insulin resistance (IR), which is closely associated with the pathophysiology of polycystic ovary syndrome (PCOS). Abnormalities in adipocyte function have been identified in PCOS and potentially contribute to lower adiponectin concentrations. Objective: Our objective was to determine which variables in plasma and adipose tissue influence HMW adiponectin in a well characterized cohort of women with PCOS. Design: This was a cross-sectional study. Settings and Participants: A teaching hospital. Women with PCOS (n = 98) and body mass index (BMI)-matched controls (n = 103) (including 68 age-, BMI-, and IR-matched pairs). Interventions: A standard 75-g oral glucose tolerance test was performed for each participant. Subcutaneous adipose tissue samples were taken by needle biopsy for a subset of PCOS women (n = 9) and controls (n = 8). Main Outcome Measures: Serum levels of HMW adiponectin and their relation to indices of insulin sensitivity, body composition, and circulating androgens as well as adipose tissue expression levels of ADIPOQ, TNFalpha, PPARgamma, and AR were assessed. Results: HMW adiponectin was significantly lower in women with PCOS compared with both BMI- and BMI- and IR-matched controls (P = 0.009 and P = 0.027, respectively). Although BMI and IR were the main predictors of HMW adiponectin, an interaction between waist to hip ratio and plasma testosterone contributed to its variance (P = 0.026). Adipose tissue gene expression analysis demonstrated that AR and TNFalpha (P = 0.008 and P = 0.035, respectively) but not ADIPOQ mRNA levels were increased in PCOS compared with controls. Conclusions: HMW adiponectin is selectively reduced in women with PCOS, independent of BMI and IR. Gene expression analysis suggests that posttranscriptional\\/translational modification contributes to reduced HMW adiponectin in PCOS.

  17. Effect of berberine on the ratio of high-molecular weight adiponectin to total adiponectin and adiponectin receptors expressions in high-fat diet fed rats.

    Science.gov (United States)

    Wu, Yue-Yue; Zha, Ying; Liu, Jun; Wang, Fang; Xu, Jiong; Chen, Zao-Ping; Ding, He-Yuan; Sheng, Li; Han, Xiao-Jie

    2016-11-17

    To assess the effects of berberine (BBR) on high-molecular weight (HMW) adiponectin and adiponectin receptors (adipoR1/adipoR2) expressions in high-fat (HF) diet fed rats. Forty Wistar male rats were randomly assigned into a normal diet fed group and three HF diet (fat for 45% calories) fed groups (n=10 for each group). All rats underwent 12 weeks of feeding. After 4 weeks feeding, rats in the two of three HF diet fed groups were treated with 150 mg·kg -1 ·day -1 BBR (HF+LBBR group) and 380 mg·kg -1 ·day -1 BBR (HF+HBBR group) by gavage once a day respectively for the next 8 weeks while the rats in other groups treated with vehicle (NF+Veh and HF+Veh). Body weight and food intake were observed and recorded on daily basis. At the end of 12 weeks, the blood, liver, epididymal fat tissues and quadriceps femoris muscles were collected. Fasting insulin, plasma fasting glucose, serum free fatty acid (FFA), total adiponectin and HMW adiponectin levels were measured by enzyme linked immunosorbent assay method. Glucose tolerance test (GTT) and insulin tolerance test (ITT) were performed to determine the insulinsensitizing. Meanwhile the homeostasis model assessment (HOMA) method was used to determine insulin resistance (HOMA-IR). The expressions of adipoR1, adipoR2 and adenosine monophophate activated protein kinase (AMPK) phosphorylation level in skeletal muscle and liver tissue were detected by Western blot. Liver and kidney toxicity were evaluated during treatment. The body weight of rats in high- or low-dose BBR group reduced as well as HOMA-IR, FFA concentrations and fasting insulin levels decreased compared with HF+Veh group (Pinsulin resistance by increasing the expression of adiponectin receptors and the ratio of HMW to total adiponectin.

  18. Serum high molecular weight complex of adiponectin correlates better with glucose tolerance than total serum adiponectin in Indo-Asian males.

    Science.gov (United States)

    Fisher, F F M; Trujillo, M E; Hanif, W; Barnett, A H; McTernan, P G; Scherer, P E; Kumar, S

    2005-06-01

    It is well established that total systemic adiponectin is reduced in type 2 diabetic subjects. To date most studies have been concerned with the singular full-length protein or proteolytically cleaved globular domain. It is, however, apparent that the native protein circulates in serum as a lower molecular weight hexamer and as larger multimeric structures of high molecular weight (HMW). In this study we address the clinical significance of each form of the protein with respect to glucose tolerance. Serum was obtained from 34 Indo-Asian male subjects (BMI 26.5+/-3.1; age 52.15+/-10.14 years) who had undertaken a 2-h oral glucose tolerance test. An aliquot of serum was fractionated using velocity sedimentation followed by reducing SDS-PAGE. Western blots were probed for adiponectin, and HMW adiponectin as a percentage of total adiponectin (percentage of higher molecular weight adiponectin [S(A)] index) was calculated from densitometry readings. Total adiponectin was measured using ELISA; leptin, insulin and IL-6 were determined using ELISA. Analysis of the cohort demonstrated that total adiponectin (r = 0.625, p = 0.0001), fasting insulin (r = -0.354, p = 0.040) and age (r = 0.567, p = 0.0001) correlated with S(A). S(A) showed a tighter, inverse correlation with 2-h glucose levels (r = -0.58, p = 0.0003) than total adiponectin (r = -0.38, p = 0.0001). This study demonstrates the importance of the S(A) index as a better determinant of glucose intolerance than measurements of total adiponectin. Our findings suggest that HMW adiponectin is the active form of the protein.

  19. Simultaneous detection of low and high molecular weight carbonylated compounds derived from lipid peroxidation by electrospray ionization-tandem mass spectrometry.

    Science.gov (United States)

    Milic, Ivana; Hoffmann, Ralf; Fedorova, Maria

    2013-01-02

    Reactive oxygen species (ROS) and other oxidative agents such as free radicals can oxidize polyunsaturated fatty acids (PUFA) as well as PUFA in lipids. The oxidation products can undergo consecutive reactions including oxidative cleavages to yield a chemically diverse group of products, such as lipid peroxidation products (LPP). Among them are aldehydes and ketones ("reactive carbonyls") that are strong electrophiles and thus can readily react with nucleophilic side chains of proteins, which can alter the protein structure, function, cellular distribution, and antigenicity. Here, we report a novel technique to specifically derivatize both low molecular and high molecular weight carbonylated LPP with 7-(diethylamino)coumarin-3-carbohydrazide (CHH) and analyze all compounds by electrospray ionization-mass spectrometry (ESI-MS) in positive ion mode. CHH-derivatized compounds were identified by specific neutral losses or fragment ions. The fragment ion spectra displayed additional signals that allowed unambiguous identification of the lipid, fatty acids, cleavage sites, and oxidative modifications. Oxidation of docosahexaenoic (DHA, 22:6), arachidonic (AA, 20:4), linoleic (LA, 18:2), and oleic acids (OA, 18:1) yielded 69 aliphatic carbonyls, whose structures were all deduced from the tandem mass spectra. When four phosphatidylcholine (PC) vesicles containing the aforementioned unsaturated fatty acids were oxidized, we were able to deduce the structures of 122 carbonylated compounds from the tandem mass spectra of a single shotgun analysis acquired within 15 min. The high sensitivity (LOD ∼ 1 nmol/L for 4-hydroxy-2-nonenal, HNE) and a linear range of more than 3 orders of magnitude (10 nmol/L to 10 μmol/L for HNE) will allow further studies on complex biological samples including plasma.

  20. Increased synthesis of high-molecular-weight cPLA2 mediates early UV-induced PGE2 in human skin.

    Science.gov (United States)

    Gresham, A; Masferrer, J; Chen, X; Leal-Khouri, S; Pentland, A P

    1996-04-01

    Ultraviolet light (UV) B-induced inflammation is characterized by dramatic increases in prostaglandin E2 (PGE2) synthesis due to enhanced arachidonate deacylation from the membrane. Therefore, the effect of UV on sythesis, mass, and distribution of the high-molecular-weight phospholipase A2 (cPLA2) in cultured human keratinocytes and human skin was studied. The 105-kDa cPLA2 was demonstrated to be the critical enzyme in UV-induced PGE2 synthesis and erythema in the first 6 h postirradiation. Immunoprecipitation of 35S-labeled protein showed cPLA2 synthesis increased three- to fourfold 6 h after irradiation. Immunoprecipitated 32P-labeled cPLA2 demonstrated phosphorylation of cPLA2 was concurrently induced, suggesting that UV also activates cPLA2. This increase in cPLA2 synthesis and activation also closely correlated with increased PGE2 synthesis and [3H]arachidonic acid release and was effectively blocked by both an S-oligonucleotide antisense to cPLA2 and methyl arachidonate fluorophosphate, a specific inhibitor of cPLA2. Biopsy and histochemical examination of erythematous sites expressed increased amounts of cPLA2 whereas nonerythematous irradiated sites did not. In contrast, cyclooxygenase-1 and -2 in cultures and skin explants were unaffected 6 h post-UV, and no change in cyclooxygenase activity was observed at this time. These results suggest that increased cPLA2 synthesis occurs only when skin is exposed to UV doses that are sufficient to cause erythema and indicate expression of cPLA2 participates in acute UV inflammation.

  1. Effects of body fat on the associations of high-molecular-weight adiponectin, leptin and soluble leptin receptor with metabolic syndrome in Chinese.

    Directory of Open Access Journals (Sweden)

    Danxia Yu

    Full Text Available BACKGROUND: Little is known regarding the associations between high-molecular-weight (HMW- adiponectin, leptin and soluble leptin receptor (sOB-R and metabolic syndrome (MetS in Chinese. Also few studies elucidate the effects of inflammation and body fat mass on the relations. METHODS: Plasma HMW-adiponectin, leptin and sOB-R were measured among 1055 Chinese men and women (35∼54 yrs. Whole body and trunk fat mass were determined by Dual-energy X-ray absorptiometry. MetS was defined by the updated NCEP/ATPIII criterion for Asian-Americans. RESULTS: HMW-adiponectin was inversely associated with MetS in multivariate model including fat mass index (FMI, inflammatory markers, leptin and sOB-R (OR in the highest quartile= 0.30, 95%CI 0.18∼0.50, P<.0001. Plasma sOB-R was also inversely associated with MetS independent of body fatness and inflammatory markers, whereas the association was somewhat attenuated after adjusting HMW-adiponectin (OR for the highest quartile = 0.78, 95%CI 0.47∼1.32, P = 0.15. In contrast, leptin was associated with increased odds of MetS independent of inflammatory markers, HMW-adiponectin, and sOB-R (OR for the highest quartile= 2.64, 95%CI 1.35∼5.18, P = 0.006, although further adjustment for FMI abolished this association. CONCLUSIONS: HMW-adiponectin exhibited strong inverse associations with MetS independent of body composition, inflammation, leptin and sOB-R; while the associations of leptin and sOB-R were largely explained by fat mass or HMW-adiponectin, respectively.

  2. The relative importance of competing pathways for the formation of high-molecular-weight peroxides in the ozonolysis of organic aerosol particles

    Directory of Open Access Journals (Sweden)

    M. Mochida

    2006-01-01

    Full Text Available High-molecular-weight (HMW organic compounds are an important component of atmospheric particles, although their origins, possibly including in situ formation pathways, remain incompletely understood. This study investigates the formation of HMW organic peroxides through reactions involving stabilized Criegee intermediates (SCI's. The model system is methyl oleate (MO mixed with dioctyl adipate (DOA and myristic acid (MA in submicron aerosol particles, and Criegee intermediates are formed by the ozonolysis of the double bond in methyl oleate. An aerosol flow tube coupled to a quadrupole aerosol mass spectrometer (AMS is employed to determine the relative importance of different HMW organic peroxides following the ozonolysis of different mixing mole fractions of MO in DOA and MA. Possible peroxide products include secondary ozonides (SOZ's, α-acyloxyalkyl hydroperoxides and α-acyloxyalkyl alkyl peroxides (αAAHP-type compounds, diperoxides, and monoperoxide oligomers. Of these, the AMS data identify two SOZ's as major HMW products in the ozonolysis of pure methyl oleate as well as in an inert matrix of DOA to as low as 0.04 mole fraction MO. In comparison, in mixed particles of MO and MA, αAAHP-type compounds form in high yields for MO mole fractions of 0.5 or less, suggesting that SCI's efficiently attack the carboxylic acid group of myristic acid. The reactions of SCI's with carboxylic acid groups to form αAAHP-type compounds therefore compete with those of SCI's with aldehydes to form SOZ's, provided that both types of functionalities are present at significant concentrations. The results therefore suggest that SCI's in atmospheric particles contribute to the transformation of carboxylic acids and other protic groups into HMW organic peroxides.

  3. Dynamics of Nampt/visfatin and high molecular weight adiponectin in response to oral glucose load in obese and lean women.

    Science.gov (United States)

    Unlütürk, Uğur; Harmanci, Ayla; Yildiz, Bülent Okan; Bayraktar, Miyase

    2010-04-01

    High molecular weight adiponectin (HMWA) is the active circulating form of adiponectin. Nampt/visfatin is the enzyme secreted from adipocytes in an active form and is one of the putative regulators of insulin secretion. To investigate the dynamics of total adiponectin (TA), HMWA and Nampt/visfatin in obese and lean women during oral glucose tolerance test (OGTT). We studied normal glucose-tolerant (NGT), age-matched, 30 obese and 30 lean women. All subjects underwent a standard 75 g, 2-h OGTT, and area under the curve (AUC) during OGTT for glucose, insulin, Nampt/visfatin, TA and HMWA was calculated. Body fat mass was assessed by bioimpedance analysis. Results Obese women had significantly higher basal and AUC values for insulin and Nampt/visfatin, whereas basal and AUC-HMWA were significantly lower in this group. Alternatively, obese and lean groups had similar basal and AUC values for glucose and TA. Basal insulin levels were negatively correlated with HMWA levels, but not with basal Nampt/visfatin. AUC-insulin was correlated positively with AUC-visfatin, and negatively with AUC-HMWA. Total and truncal body fat mass showed positive correlation with basal and AUC-visfatin, and negative correlation with basal and AUC-HMWA. In the NGT state, obese women have higher Nampt/visfatin and lower HMWA levels, both basally and in response to oral glucose challenge. The dynamics of Nampt/visfatin and HMWA during OGTT appear to be linked with insulin and adiposity. Counter-regulatory adaptations in HMWA and Nampt/visfatin might have an impact on suggested adipoinsular axis, contributing to maintenance of normal glucose tolerance.

  4. Formation of high-molecular-weight compounds via the heterogeneous reactions of gaseous C8-C10 n-aldehydes in the presence of atmospheric aerosol components

    Science.gov (United States)

    Han, Yuemei; Kawamura, Kimitaka; Chen, Qingcai; Mochida, Michihiro

    2016-02-01

    A laboratory study on the heterogeneous reactions of straight-chain aldehydes was performed by exposing n-octanal, nonanal, and decanal vapors to ambient aerosol particles. The aerosol and blank filters were extracted using methanol. The extracts were nebulized and the resulting compositions were examined using a high-resolution time-of-flight aerosol mass spectrometer. The mass spectral analysis showed that the exposures of the aldehydes to aerosol samples increased the peak intensities in the high mass range. The peaks in the mass spectra of the aerosol samples after exposure to different aldehydes were characterized by a homologous series of peak shifts due to the addition of multiple CH2 units. This result is explained by the formation of high-molecular-weight (HMW) compounds that contain single or multiple aldehyde moieties. The HMW fragment peaks for the blank filters exposed to n-aldehydes were relatively weak, indicating an important contribution from the ambient aerosol components to the formation of the HMW compounds. Among the factors affecting the overall interaction of aldehydes with atmospheric aerosol components, gas phase diffusion possibly limited the reactions under the studied conditions; therefore, their occurrence to a similar degree in the atmosphere is not ruled out, at least for the reactions involving n-nonanal and decanal. The major formation pathways for the observed HMW products may be the self-reactions of n-aldehydes mediated by atmospheric aerosol components and the reactions of n-aldehydes with organic aerosol components. The observed formation of HMW compounds encourages further investigations into their effects on the aerosol properties as well as the organic aerosol mass in the atmosphere.

  5. Wear resistant performance of highly cross-linked and annealed ultra-high molecular weight polyethylene against ceramic heads in total hip arthroplasty.

    Science.gov (United States)

    Sato, Taishi; Nakashima, Yasuharu; Akiyama, Mio; Yamamoto, Takuaki; Mawatari, Taro; Itokawa, Takashi; Ohishi, Masanobu; Motomura, Goro; Hirata, Masanobu; Iwamoto, Yukihide

    2012-12-01

    The purpose of this study was to examine the effects of ceramic femoral head material, size, and implantation periods on the wear of annealed, cross-linked ultra-high molecular weight polyethylene (UHMWPE) (XLPE) in total hip arthroplasty compared to non-cross-linked conventional UHMWPE (CPE). XLPE was fabricated by cross-linking with 60 kGy irradiation and annealing. Femoral heads made from zirconia and alumina ceramics and cobalt-chrome (CoCr) of 22 or 26 mm diameter were used. In this retrospective cohort study, the femoral head penetration into the cup was measured digitally on radiographs of 367 hips with XLPE and 64 hips with CPE. The average follow-up periods were 6.3 and 11.9 years, respectively. Both XLPE creep and wear rates were significantly lower than those of CPE (0.19 mm vs. 0.44 mm, 0.0001 mm/year vs. 0.09 mm/year, respectively). Zirconia displayed increased wear rates compared to alumina in CPE; however, there was no difference among head materials in XLPE (0.0008, 0.00007, and -0.009 mm/year for zirconia, alumina, and CoCr, respectively). Neither head size or implantation period impacted XLPE wear. In contrast to CPE, XLPE displayed low wear rates surpassing the effects of varying femoral head material, size, implantation period, and patient demographics. Further follow-up is required to determine the long-term clinical performance of the annealed XLPE. Copyright © 2012 Orthopaedic Research Society.

  6. Comparison of novel ultra-high molecular weight polyethylene tape versus conventional metal wire for sublaminar segmental fixation in the treatment of adolescent idiopathic scoliosis.

    Science.gov (United States)

    Takahata, Masahiko; Ito, Manabu; Abumi, Kuniyoshi; Kotani, Yoshihisa; Sudo, Hideki; Ohshima, Shigeki; Minami, Akio

    2007-08-01

    Retrospective study. To compare the surgical outcomes of posterior translational correction and fusion using hybrid instrumentation systems with either sublaminar Nesplon tape or sublaminar metal wire to treat adolescent idiopathic scoliosis (AIS). Nesplon tape, which consists of a thread of ultra-high molecular weight polyethylene fibers, has advantages over metal wire: (1) its soft and flexible properties avoid neural damage and (2) its flat configuration avoids focal distribution of the stresses to lamina; however, the efficacy of Nesplon tape in the correction of spinal deformity is as yet, still unclear. Thirty AIS patients at a single institution underwent posterior correction and fusion using hybrid instrumentation containing hook, pedicle screw, and either sublaminar polyethylene taping (15) or sublaminar metal wiring (15). Patients were evaluated preoperatively, immediately after surgery, and at a 2-year follow-up according to the radiographic changes in curve correction, operating time, intraoperative blood loss, complications, and the Scoliosis Research Society patient questionnaire (SRS-24) score. The average correction rate was 63.0% in the Nesplon tape group and 59.9% in the metal wire group immediately after surgery (P = 0.62). Fusion was obtained in all the patients without significant correction loss in both groups. There was no significant difference in operative time, intraoperative blood loss, and postoperative SRS-24 scores between the 2 groups. Complications were superficial skin infection in a single patient in the Nesplon tape group, and transient sensory disturbance in 1 patient and temporal superior mesenteric artery syndrome in another patient in the metal wire group. The efficacy of Nesplon tape in correction of deformity is equivalent to that of metal wire, and fusion was completed without significant correction loss. The soft and flexible properties and flat configuration of Nesplon tape make this a safe application for the treatment

  7. C-reactive protein, high-molecular-weight adiponectin and development of metabolic syndrome in the Japanese general population: a longitudinal cohort study.

    Directory of Open Access Journals (Sweden)

    Yoshifumi Saisho

    Full Text Available AIMS: To clarify predictive values of C-reactive protein (CRP and high-molecular-weight (HMW adiponectin for development of metabolic syndrome. RESEARCH DESIGN AND METHODS: We conducted a prospective cohort study of Japanese workers who had participated in an annual health checkup in 2007 and 2011. A total of 750 subjects (558 men and 192 women, age 46±8 years who had not met the criteria of metabolic syndrome and whose CRP and HMW-adiponectin levels had been measured in 2007 were enrolled in this study. Associations between CRP, HMW-adiponectin and development of metabolic syndrome after 4 years were assessed by logistic regression analysis and their predictive values were compared by receiver operating characteristic analysis. RESULTS: Among 750 subjects, 61 (8.1% developed metabolic syndrome defined by modified National Cholesterol Education Program Adult Treatment Panel III (NCEP-ATP III criteria and 53 (7.1% developed metabolic syndrome defined by Japan Society for the Study of Obesity (JASSO in 2011. Although CRP and HMW-adiponectin were both significantly correlated with development of metabolic syndrome, multivariate logistic regression analysis revealed that HMW-adiponectin but not CRP was associated with metabolic syndrome independently of BMI or waist circumference. Adding these biomarkers to BMI or waist circumference did not improve the predictive value for metabolic syndrome. CONCLUSION: Our findings indicate that the traditional markers of adiposity such as BMI or waist circumference remain superior markers for predicting metabolic syndrome compared to CRP, HMW-adiponectin, or the combination of both among the Japanese population.

  8. Plasma parameters effects on the properties, aging and stability behaviors of allylamine plasma coated ultra-high molecular weight polyethylene (UHMWPE) films

    Energy Technology Data Exchange (ETDEWEB)

    Aziz, Gaelle, E-mail: gaelle.aziz@ugent.be; Thukkaram, Monica; De Geyter, Nathalie; Morent, Rino

    2017-07-01

    Highlights: • Medium to atmospheric pressure DBD is used to deposit amino rich films. • Process parameters affect the films’ surface chemical and physical properties. • High deposition rates can be reached by varying the power and/or monomer flow rate. • High amino selectivity (NH{sub 2}/N in %) is obtained at low powers and high monomer concentration. • Aging and stability behaviors of the deposited coatings can be controlled by carefully choosing the plasma parameters. - Abstract: In this work, a dielectric barrier discharge (DBD) operated at medium to atmospheric pressure has been used for the deposition of thin polyallylamine (PAA) films on ultra-high molecular weight polyethylene (UHMWPE) substrates. The effect of treatment time (1–5 min), discharge power (5.7–24.0 W), monomer concentration (1–2 g/h) and pressure (10–100 kPa) on the films properties, aging and stability behaviors have been investigated. The used characterization techniques are X-ray photoelectron spectroscopy, water contact angle and optical reflectance spectroscopy. In this paper, it is shown that plasma treatment time does not affect the coatings chemistry; whereas plasma power, monomer concentration and pressure control the coatings properties. It is also shown that the deposition rate of the deposited films changes with varying W/FM values. At low W/FM values, high deposition rates of up to 2 nm/s are observed. Plasma treatments were also characterized by their amino efficiency ([NH{sub 2}]/[C] in %) and amino selectivity ([NH{sub 2}]/[N] in %). Depending on the used parameters, these varied between 12.3% and 20% and between 71.2% and 91.1%, respectively. For the aging study, coatings that preserved most of their hydrophilicity were obtained at power ≤11.3 W, monomer concentration ≥1.5 g/h and pressure ≥50 kPa. For the stability study, coatings that showed the highest [N] (%) and lowest percentage of thickness decrease were obtained at ≤2 min, 24.0 W, 1 g/h and

  9. Measurements of free radical in vitamin E-doped ultra-high molecular weight polyethylene: Dependence on materials processing and irradiation environments

    Energy Technology Data Exchange (ETDEWEB)

    Ridley, M.D. [Department of Physics, Biomaterials Research Laboratory, University of Memphis, 216 Manning Hall, Memphis, TN 38152 (United States); Jahan, M.S. [Department of Physics, Biomaterials Research Laboratory, University of Memphis, 216 Manning Hall, Memphis, TN 38152 (United States)], E-mail: mjahan@memphis.edu

    2007-12-15

    Ultra-high molecular weight polyethylene (UHMWPE), doped with vitamin E ({alpha}-tocopherol ({alpha}-T)), was irradiated with gamma rays in nitrogen (N{sub 2}) or air, and the resulting free radicals were detected in air using an electron spin resonance (ESR) technique. Two groups of samples were investigated. In one group, samples were prepared from blends of {alpha}-T (20 wt%) and UHMWPE powder (PPE-{alpha}-T) and, in the other, from compression molded blocks (CMPE-{alpha}-T). The CMPE-{alpha}-T blocks contained 0% (control), 0.5%, 1.0%, 10.0%, 15.0%, 20.0% and 25.0% {alpha}-T by weight. When irradiation was performed in air, the ESR spectrum of powder samples showed the presence of only vitamin E radical (tocopheroxyl, {alpha}-T-O{sup {center_dot}}), and there was no detectable signal due to PE radicals (alkyl/allyl). Most likely, all PE radicals were quenched by vitamin E during irradiation in air. However, when irradiation was performed in N{sub 2}, composite ESR spectra showed the presence of both PE and {alpha}-T-O{sup {center_dot}} radicals. Compared to the control (PPE, 0% {alpha}-T) PE radicals in PPE-20% {alpha}-T were found to be significantly reduced or quenched by {alpha}-T. The presence of {alpha}-T in powder samples, however, did not affect the long-term (71 days in this study) oxidation behavior of the PE radicals. Compression molded samples, with and without {alpha}-T, produced identical ESR spectra irrespective of their irradiation environment N{sub 2} or air. However, radical concentration, measured immediately after irradiation, was found to be an order of magnitude less in CMPE-{alpha}-T than in the control (CMPE-0% {alpha}-T). They also evidenced identical structural changes in the respective ESR spectra during subsequent oxidation for 24 days in open air. These observations suggest that {alpha}-T can effectively quench a significant fraction of PE radicals during irradiation, but has no measurable effect on subsequent reactions. No

  10. Measurements of Free Radical in Vitamin E-Doped Ultra-High Molecular Weight Polyethylene: Dependence on Materials Processing and Irradiation Environments

    International Nuclear Information System (INIS)

    Ridley, M. D.

    2006-01-01

    In an effort to combat oxidation of the load-bearing, polyethylene (PE) components of total hip- and knee-joint replacement devices, antioxidant such as vitamin E (α-Tocopherol (α-T)) has been introduced into polymer matrix. In this study we investigated effect of α-T on free radicals in medical grade ultra-high molecular weight polyethylene (UHMWPE), GUR 1020 resin. Since oxidation resistance depends on radical reaction, we used electron spin resonance (ESR) technique for direct detection of free radicals before and after irradiation. High concentration (20% by vol.) of α-T was used so its ESR signal (due to α-T-O degree radical) could be distinguished among the overwhelming signals due to PE radicals. Two groups of samples were investigated. In one group, samples were prepared from blends of α-T and UHMWPE powder (α-T-P), and in the second group, from compression molded blocks (α-T-B). In each group, samples were γ-irradiated in sealed packages filled with N 2 , or in open air, and free radicals were measured in open air environment as a function of time. Also included in this study were α-T, and powder resin and compression molded blocks without any α-T. Following irradiation in air, α-T-P and α-T produced identical ESR spectra showing characteristic feature of α-T-O degree radical. Absence of PE radicals in the ESR signals suggests quenching/repairing of PE radicals by α-T in presence of oxygen. However, when irradiation was performed in N 2 , ESR signals of α-T-P exhibited superimposed resonance lines due to PE and α-T-O degree radicals. Furthermore, presence of a-T or α-T-O degree radicals did not prevent subsequent oxidation of PE radicals in producing stable, oxygen-induced radicals (OIR). Post-irradiation oxidation index measured by FTIR, however, did not show any difference between these samples. Compression molded samples, with or without α-T, produced ESR spectra showing features characteristics of PE radicals only, and there was no

  11. Processing of high-molecular-weight form adrenocorticotropin in human adrenocorticotropin-secreting tumor cell line (DMS-79) after transfection of prohormone convertase 1/3 gene.

    Science.gov (United States)

    Tateno, T; Kato, M; Tani, Y; Yoshimoto, T; Oki, Y; Hirata, Y

    2010-02-01

    Ectopic ACTH-producing tumors preferentially secrete biologically inactive ACTH precursors and ACTH-related fragments. DMS-79 is known to secrete unprocessed high-molecular-weight (HMW) form ACTH. To determine whether prohormone convertase (PC) 1/3 is involved in the abnormal processing of proopiomelanocortin (POMC), we studied whether PC1/3 and 2 genes are expressed in DMS-79, and whether overexpression of PC1/3 gene affects POMC processing pattern. Steady-state mRNA levels of PC1/3 and 2 were determined by real-time RT-PCR. Molecular weights of ACTH-related peptides were determined by chromatographical analyses coupled with ACTH and beta-endorphin (beta-END) radioimmunoassays. PC1/3 gene was transfected into DMS-79 by retrovirus transduction using pMX-IP vector encoding PC1/3 cDNA. The steady-state mRNA levels of PC1/3 and 2 in DMS-79 were lower than those in ACTH-secreting and nonfunctioning pituitary tumors. DMS-79 predominantly secreted HMW form with both ACTH and beta-END immunoreactivities by size-exclusion chromatography. After purification by immunoaffinity chromatography with anti-ACTH antibody, the apparent molecular weight of HMW form ACTH was estimated to be 16 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with silver staining. After retroviral transfection of PC1/3 cDNA into DMS-79 and puromycin selection, PC1/3 stably-expressing cell line (DMS-79T) secreted two immunoreactive ACTH components, a major one coeluting with ACTH(1-39) and a minor one as a HMW form as well as two beta- END immunoreactive components coeluting with beta-lipotropic hormone and beta-END, respectively. Thus, we have established PC1/3 stably-expressing cell line (DMS-79T) capable of proteolytically processing ACTH precursor molecule(s) into mature ACTH and beta-END.

  12. Surface modification of vascular endothelial growth factor-loaded silk fibroin to improve biological performance of ultra-high-molecular-weight polyethylene via promoting angiogenesis

    Directory of Open Access Journals (Sweden)

    Ai C

    2017-10-01

    Full Text Available Chengchong Ai, Dandan Sheng, Jun Chen, Jiangyu Cai, Siheng Wang, Jia Jiang, Shiyi Chen Department of Sports Medicine, Huashan Hospital, Fudan University, Shanghai, People’s Republic of China Abstract: Ultra-high-molecular-weight polyethylene (UHMWPE has been applied in orthopedics, as the materials of joint prosthesis, artificial ligaments, and sutures due to its advantages such as high tensile strength, good wear resistance, and chemical stability. However, postoperative osteolysis induced by UHMWPE wear particles and poor bone–implant healing interface due to scarcity of osseointegration is a significant problem and should be solved imperatively. In order to enhance its affinity to bone tissue, vascular endothelial growth factor (VEGF was loaded on the surface of materials, the loading was performed by silk fibroin (SF coating to achieve a controlled-release delivery. Several techniques including field emission scanning electron microscopy (FESEM and attenuated total reflectance (ATR-Fourier transform infrared (FTIR and water contact angle measurement were used to validate the effectiveness of introduction of SF/VEGF. The result of ELISA demonstrated that the release of VEGF was well maintained up to 4 weeks. The modified UHMWPE was evaluated by both in vitro and in vivo experiments. According to the results of FESEM and cell counting kit-8 (CCK-8 assay, bone marrow mesenchymal stem cells cultured on the UHMWPE coated with SF/VEGF and SF exhibited a better proliferation performance than that of the pristine UHMWPE. The model rabbit of anterior cruciate ligament reconstruction was used to observe the graft–bone healing process in vivo. The results of histological evaluation, microcomputed tomography (micro-CT analysis, and biomechanical tests performed at 6 and 12 weeks after surgery demonstrated that graft–bone healing could be significantly improved due to the effect of VEGF on angiogenesis, which was loaded on the surface by SF

  13. Chemical modification of high molecular weight polyethylene through gamma radiation for biomaterials applications; Modificacao quimica de polietileno de alto peso molecular atraves de radiacao gama para aplicacao em biomateriais

    Energy Technology Data Exchange (ETDEWEB)

    Raposo, Matheus P.; Rocha, Marisa C.G., E-mail: matheusmerlim@hotmail.com [Universidade Estado do Rio de Janeiro (UERJ), Nova Friburgo, RJ (Brazil). Instituto Politecnico

    2015-07-01

    Ultra high molecular weight polyethylene has been used in the medical field due to its high mechanical properties compared to the other polymers. Its main application is in the development of orthopedic implants, which requires high resistance to abrasion. One of the most used methods is the introduction of crosslinks in the polymer through gamma irradiation. In order to prevent oxidation reactions, studies have been developed using tacoferol (vitamin E) as an antioxidant for the material. The ascorbic acid (vitamin C), however, has been appointed as a viable alternative for vitamin E. In this work, a high molecular weight polyethylene grade (HMWPE) and polyethylene samples formulated with vitamin C were submitted to gamma radiation. Thermodynamic-mechanical methods and gel content determinations were used to characterize the samples obtained. The sample containing 1% of vitamin C and irradiated with 50 KGy of gamma radiation presented the highest content of crosslinks. (author)

  14. Long term efficacy and safety of a combined low and high molecular weight hyaluronic acid in the treatment of osteoarthritis of the knee

    Directory of Open Access Journals (Sweden)

    Robert J. Petrella

    2011-03-01

    Full Text Available The primary objective was to determine the efficacy of intraarticular combined hyaluronic acid versus placebo in patients with grade 1-3 medial compartment osteoarthritis of the knee as evaluated through the self-paced 40 m walking pain visual analog scale (VAS at week 16, 52 and 104. Secondary objectives included pain at rest: a 10 cm VAS, patient global satisfaction using a 5-point numerical scale, consumption of concomitant medications, patients with <45 mm pain at followup 52 and 104 weeks. Safety was determined through the number of recorded adverse events. The study was designed as a prospective, randomized, double-blind, placebo controlled and comparative study. 200 patients were randomized in a 1:1 ratio to one of four treatment groups, to receive 3 weekly intra-articular injections of either: DMW (combined HA of different molecular weight and concentrations; HMW (high molecular weight HA; LMW (low molecular weight HA; PL (placebo, saline. Patients were assessed baseline and at week 16, 52, 104. Analyses were conducted using sigma stat (SPSS Inc., Chicago, Illinois and Microsoft Excel (Microsoft Corp, Redmond, Washington. Significance was established at P<0.05. Analysis of variance with repeated measures and c2 tests were used to test for differences from baseline characteristics of the group among the primary and secondary outcomes at each injection series interval. At 16, 52 and 104 weeks respectively, walking VAS pain was significantly improved in all treatment groups vs. Placebo: DMW (89.3%, P<0.001; 87.4%, P<0.001; 88.1%, P<0.001; LMW (81.3%, P<0.001; 78.2%, P<0.001; 77.0%, P<0.001 and HMW (79.1%, P<0.001; 81.1%, P<0.001; 79.4%, P<0.001. At 52 weeks, 8 patients in DMW group had resting VAS <45 mm. DMW had lower (62 mm, P <0.001 compared to LMW (76 mm and HMW (88 mm VAS at rest. Similar differences were observed for walking VAS 39, 41 and 43 (DMW, LMW, HMW received repeat injections. At 104 weeks, these differences were similar. DMW

  15. Transdermal delivery of relatively high molecular weight drugs using novel self-dissolving microneedle arrays fabricated from hyaluronic acid and their characteristics and safety after application to the skin.

    Science.gov (United States)

    Liu, Shu; Jin, Mei-na; Quan, Ying-shu; Kamiyama, Fumio; Kusamori, Kosuke; Katsumi, Hidemasa; Sakane, Toshiyasu; Yamamoto, Akira

    2014-02-01

    The purpose of this study was to develop novel dissolving microneedle arrays fabricated from hyaluronic acid (HA) as a material and to improve the transdermal permeability of relatively high molecular weight drugs. In this study, fluorescein isothiocyanate-labeled dextran with an average molecular weight of 4kDa (FD4) was used as a model drug with a relatively high molecular weight. The microneedle arrays significantly increased transepidermal water loss (TEWL) and reduced transcutaneous electrical resistance (TER), indicating that they could puncture the skin and create drug permeation pathways successfully. Both TEWL and TER almost recovered to baseline levels in the microneedle array group, and relatively small pathways created by the microneedles rapidly recovered as compared with those created by a tape stripping treatment. These findings confirmed that the microneedle arrays were quite safe. Furthermore, we found that the transdermal permeability of FD4 using the microneedle arrays was much higher than that of the FD4 solution. Furthermore, we found that the microneedle arrays were much more effective for increasing the amount of FD4 accumulated in the skin. These findings indicated that using novel microneedle arrays fabricated from HA is a very useful and effective strategy to improve the transdermal delivery of drugs, especially relatively high molecular weight drugs without seriously damaging the skin. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. Understanding the link between GMP and dough: from glutenin particles in flour towards developed dough

    NARCIS (Netherlands)

    Don, C.; Lichtendonk, W.J.; Plijter, J.J.; Hamer, R.J.

    2003-01-01

    Clear correlations exist for glutenin macropolymer (GMP) quantity and rheological properties vs. wheat quality and dough rheological properties, but real insight in understanding these links is still missing. The observation that GMP consists of glutenin particles opens up new possibilities to

  17. Understanding the link between GMP and dough: From glutenin particles in flour towards developed dough

    NARCIS (Netherlands)

    Don, C.; Lichtendonk, W.J.; Plijter, J.J.; Hamer, R.J.

    2003-01-01

    Clear correlations exist for glutenin macropolymer (GMP) quantity and rheological properties vs. wheat quality and dough rheological properties, but real insight in understanding these links is still missing. The observation that GMP consists of glutenin particles opens up new possibilities to

  18. Marker-Assisted Selection for Recognizing Wheat Mutant Genotypes Carrying HMW Glutenin Alleles Related to Baking Quality

    Directory of Open Access Journals (Sweden)

    Mohammad Javad Zamani

    2014-01-01

    Full Text Available Allelic diversity of HMW glutenin loci in several studies revealed that allelic combinations affect dough quality. Dx5 + Dy10 subunits are related to good baking quality and Dx2 + Dy12 are related to undesirable baking quality. One of the most regular methods to evaluate the baking quality is SDS-PAGE which is used to improve baking quality labs. Marker-assisted selection is the method which can recognize the alleles related to baking quality and this method is based on polymerase chain reaction. 10 pairs of specific primers related to Dx2, Dx2.1, Dx5, Dy10, and Dy12 subunits were used for recognizing baking quality of some wheat varieties and some mutant genotypes. Only 5 pairs of them could show the specific bands. All subunits were recognized by the primers except Dx2.1. Some of the primers were extracted from previous studies and the others were designed based on D genome subunits of wheat. SDS-PAGE method accomplished having confidence in these marker’s results. To realize the effect of mutation, seed storage proteins were measured. It showed that mutation had effect on the amount of seed storage protein on the mutant seeds (which showed polymorphism.

  19. Marker-assisted selection for recognizing wheat mutant genotypes carrying HMW glutenin alleles related to baking quality.

    Science.gov (United States)

    Zamani, Mohammad Javad; Bihamta, Mohammad Reza; Naserian Khiabani, Behnam; Tahernezhad, Zahra; Hallajian, Mohammad Taher; Shamsi, Marzieh Varasteh

    2014-01-01

    Allelic diversity of HMW glutenin loci in several studies revealed that allelic combinations affect dough quality. Dx5 + Dy10 subunits are related to good baking quality and Dx2 + Dy12 are related to undesirable baking quality. One of the most regular methods to evaluate the baking quality is SDS-PAGE which is used to improve baking quality labs. Marker-assisted selection is the method which can recognize the alleles related to baking quality and this method is based on polymerase chain reaction. 10 pairs of specific primers related to Dx2, Dx2.1, Dx5, Dy10, and Dy12 subunits were used for recognizing baking quality of some wheat varieties and some mutant genotypes. Only 5 pairs of them could show the specific bands. All subunits were recognized by the primers except Dx2.1. Some of the primers were extracted from previous studies and the others were designed based on D genome subunits of wheat. SDS-PAGE method accomplished having confidence in these marker's results. To realize the effect of mutation, seed storage proteins were measured. It showed that mutation had effect on the amount of seed storage protein on the mutant seeds (which showed polymorphism).

  20. Double-blind, placebo-controlled immunotherapy with mixed grass-pollen allergoids. IV. Comparison of the safety and efficacy of two dosages of a high-molecular-weight allergoid.

    Science.gov (United States)

    Bousquet, J; Hejjaoui, A; Soussana, M; Michel, F B

    1990-02-01

    Specific immunotherapy is still widely used in grass-pollen allergy, but its side effects may limit its use. We tested the safety and efficacy of a formalinized high-molecular-weight allergoid prepared from a mixed grass-pollen extract with two injection schedules in a double-blind, placebo-controlled study. Eighteen patients received placebo, 19 received the low-dose schedule (maximal dose: 2000 PNU) and 20 received the high-dose schedule (maximal dose: 10,000 PNU). Only one patient presented a systemic reaction of moderate severity for a dose of 1200 PNU. Before the onset of the pollen season, patients had a nasal challenge with orchard grass-pollen grains, a skin test titration, and the titration of serum-specific IgG. Both groups of patients presented a significant reduction in nasal and skin sensitivities and a significant increase in IgG compared to placebo. Symptoms and medications for rhinitis and asthma were studied during the season, and both groups receiving allergoids had a significant reduction of symptom-medication scores for nasal and bronchial symptoms. There was a highly significant correlation between nasal symptom-medication scores during the season and the results of nasal challenges. High-molecular-weight allergoids are safe and effective.

  1. Total and high molecular weight (HMW) adiponectin levels and measures of glucose and lipid metabolism following pioglitazone treatment in a randomized placebo-controlled study in polycystic ovary syndrome

    DEFF Research Database (Denmark)

    Glintborg, Dorte; Frystyk, Jan; Højlund, Kurt

    2008-01-01

    and controls and examined possible mechanisms for increased insulin sensitivity during pioglitazone treatment. STUDY SUBJECTS: Thirty PCOS patients randomized to pioglitazone, 30 mg/day, or placebo for 16 weeks and 14 weight-matched healthy females were studied. DESIGN: Total and HMW adiponectin levels were...... measured, and euglycaemic hyperinsulinaemic clamps and indirect calorimetry were performed. Delta-values denoted changes during pioglitazone treatment (16 weeks--basal). RESULTS: Pretreatment adiponectin levels were decreased in PCOS patients vs. controls (P ...OBJECTIVE: Recent studies suggested that the effect of adiponectin on insulin-stimulated glucose metabolism is mediated primarily by the high molecular weight (HMW) form of adiponectin. In the present study we evaluated total and HMW adiponectin in polycystic ovary syndrome (PCOS) patients...

  2. The circulating concentration and ratio of total and high molecular weight adiponectin in post-menopausal women with and without osteoporosis and its association with body mass index and biochemical markers of bone metabolism.

    Science.gov (United States)

    Sodi, R; Hazell, M J; Durham, B H; Rees, C; Ranganath, L R; Fraser, W D

    2009-09-01

    There is increasing evidence suggesting that adiponectin plays a role in the regulation of bone metabolism. This was a cross-sectional study of 34 post-menopausal women with and 37 without osteoporosis. All subjects had body mass index (BMI), bone mineral density (BMD), total-, high molecular weight (HMW)-adiponectin and their ratio, osteoprotegerin (OPG), a marker of bone resorption (betaCTX) and formation (P1NP) measured. We observed a positive correlation between BMI and BMD (r=0.44, plean subjects but there was no difference between those with or without osteoporosis. There were significant negative correlations between HMW/total-adiponectin ratio and BMI (r=-0.27, p=0.030) and with OPG (r=-0.44, pproduction of OPG thereby affecting osteoclasts mediated bone resorption.

  3. Allelic variation at Glu-1 loci and its association with grain quality of ...

    African Journals Online (AJOL)

    The high molecular weight glutenin subunits (HMW-GS) are important determinants of wheat flour quality. The HMW-GS patterns of the major released wheat cultivars in Sudan, in addition to some advanced lines, were analyzed using SDS-PAGE. The associations of the HMW-GS quality scores (Glu-1 QS) with the flour ...

  4. Quantitation and localisation of (in vitro) transglutaminase-catalysed glutamine hydroxylation using mass spectrometry

    NARCIS (Netherlands)

    Piersma, S.R.; Pijpekamp, A. van de; Wijngaards, G.; Gruppen, H.; Boumans, H.

    2002-01-01

    A mass spectrometric approach was chosen to quantify and localise in vitro enzymatically modified glutamine (Gln) residues in a glutamine-rich protein. This protein (named dB1), a cloned domain of the high molecular weight wheat glutenin subunit Dx5, was modified by microbial transglutaminase

  5. Journal of Genetics | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Home; Journals; Journal of Genetics. XIAOCUN ZHANG. Articles written in Journal of Genetics. Volume 96 Issue 4 September 2017 pp 563-570 RESEARCH ARTICLE. Molecular cloning and functional characterization of two novel high molecular weight glutenin subunit genes in Aegilops markgrafii · XUYE DU XIAOCUN ...

  6. Journal of Genetics | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Molecular cloning and functional characterization of two novel high molecular weight glutenin subunit genes in Aegilops markgrafii ..... Distinct classical and molecular cytogenetics of Astyanax marionae and A. fasciatus ... Clinical utility of a 377 gene custom next-generation sequencing epilepsy panel .... Contact | Site index.

  7. From image processing to classification: IV. Classification of electrophoretic patterns by neural networks and statistical methods enable quality assessment of wheat varieties for breadmaking

    DEFF Research Database (Denmark)

    Jensen, Kirsten; Kesmir, Can; Søndergaard, Ib

    1996-01-01

    The end-use quality of products made from doughs consisting of wheat flour and water is often dependent upon the storage (gluten) proteins of the grain endosperm. Today the electrophoretic patterns of the high molecular weight (HMW) glutenin subunits are used for quality selections in wheat breed...

  8. From image processing to classification: IV. Classification of electrophoretic patterns by neural networks and statistical methods enable quality assessment of wheat varieties for bread making

    DEFF Research Database (Denmark)

    Jensen, K.; Kesmir, Can; Søndergaard, Ib

    1996-01-01

    The end-use quality of products made from doughs consisting of wheat flour and water is often dependent upon the storage (gluten) proteins of the grain endosperm. Today the electrophoretic patterns of the high molecular weight (HMW) glutenin subunits are used for quality selections in wheat breed...

  9. Improved method for reliable HMW-GS identification by RP-HPLC and SDS-PAGE in common wheat cultivars

    Science.gov (United States)

    The accurate identification of alleles for high-molecular weight glutenins (HMW-GS) is critical for wheat breeding programs targeting end-use quality. RP-HPLC methods were optimized for separation of HMW-GS, resulting in enhanced resolution of 1By and 1Dx subunits. Statistically significant differe...

  10. Sintering of ultra high molecular weight polyethylene

    Indian Academy of Sciences (India)

    Unknown

    coefficient of friction, good abrasion resistance, good chemical resistance etc. It is used in ... mainly used in electrical industries, shipbuilding, coal and mining, textile ... particles deformed leading to decrease in porosity and increase in density.

  11. Solvothermal synthesis of high molecular weight dithienogermole ...

    Indian Academy of Sciences (India)

    polymers were synthesized by the solvothermal method. Optical ... compounds, especially to synthesize conjugated pol- ... analyses were performed under nitrogen flow using a .... Coffin R C, Peet J, Rogers J and Bazan G C 2009 Nat. Chem.

  12. Double-blind, placebo-controlled immunotherapy with mixed grass-pollen allergoids. III. Efficacy and safety of unfractionated and high-molecular-weight preparations in rhinoconjunctivitis and asthma.

    Science.gov (United States)

    Bousquet, J; Maasch, H J; Hejjaoui, A; Skassa-Brociek, W; Wahl, R; Dhivert, H; Michel, F B

    1989-10-01

    Specific immunotherapy with unmodified formalinized allergoids is effective in grass-pollen allergy, but systemic reactions have been observed. A high-molecular-weight formalinized allergoid (HMW-GOID) was fractionated by gel filtration, retaining molecules of greater than 85,000 daltons in the expectation of improving safety without sacrificing efficacy. HMW-GOID and unfractionated allergoid (GOID) had a similar allergenic activity assessed by RAST inhibition, but the HMW-GOID preparation was 65 times less reactive when it was tested by skin prick test than the GOID preparation. A double-blind, placebo-controlled study was carried out in grass pollen-allergic patients with placebo (14 patients), GOID (15 patients), and HMW-GOID (13 patients). An additional group of 18 patients was treated by a rush schedule with a standardized orchard grass-pollen extract. A similar mean cumulative dose was administered with both allergoids. The fractionated allergoid only elicited minor systemic reactions similar to reactions elicited by placebo, whereas 20% of patients treated by GOID and 5.5% of patients receiving the standardized extract had a severe systemic reaction. For rhinitis, conjunctivitis, and asthma, the HMW-GOID and the standardized extract had a similar efficacy, significantly greater than placebo. GOID was less effective than the other two active treatments but was significantly more effective than placebo treatment for asthma and conjunctivitis.

  13. Biotransformation of the high-molecular weight polycyclic aromatic hydrocarbon (PAH) benzo[k]fluoranthene by Sphingobium sp. strain KK22 and identification of new products of non-alternant PAH biodegradation by liquid chromatography electrospray ionization tandem mass spectrometry

    Science.gov (United States)

    Maeda, Allyn H; Nishi, Shinro; Hatada, Yuji; Ozeki, Yasuhiro; Kanaly, Robert A

    2014-01-01

    A pathway for the biotransformation of the environmental pollutant and high-molecular weight polycyclic aromatic hydrocarbon (PAH) benzo[k]fluoranthene by a soil bacterium was constructed through analyses of results from liquid chromatography negative electrospray ionization tandem mass spectrometry (LC/ESI(–)-MS/MS). Exposure of Sphingobium sp. strain KK22 to benzo[k]fluoranthene resulted in transformation to four-, three-and two-aromatic ring products. The structurally similar four-and three-ring non-alternant PAHs fluoranthene and acenaphthylene were also biotransformed by strain KK22, and LC/ESI(–)-MS/MS analyses of these products confirmed the lower biotransformation pathway proposed for benzo[k]fluoranthene. In all, seven products from benzo[k]fluoranthene and seven products from fluoranthene were revealed and included previously unreported products from both PAHs. Benzo[k]fluoranthene biotransformation proceeded through ortho-cleavage of 8,9-dihydroxy-benzo[k]fluoranthene to 8-carboxyfluoranthenyl-9-propenic acid and 9-hydroxy-fluoranthene-8-carboxylic acid, and was followed by meta-cleavage to produce 3-(2-formylacenaphthylen-1-yl)-2-hydroxy-prop-2-enoic acid. The fluoranthene pathway converged with the benzo[k]fluoranthene pathway through detection of the three-ring product, 2-formylacenaphthylene-1-carboxylic acid. Production of key downstream metabolites, 1,8-naphthalic anhydride and 1-naphthoic acid from benzo[k]fluoranthene, fluoranthene and acenaphthylene biotransformations provided evidence for a common pathway by strain KK22 for all three PAHs through acenaphthoquinone. Quantitative analysis of benzo[k]fluoranthene biotransformation by strain KK22 confirmed biodegradation. This is the first pathway proposed for the biotransformation of benzo[k]fluoranthene by a bacterium. PMID:24325265

  14. Biotransformation of the high-molecular weight polycyclic aromatic hydrocarbon (PAH) benzo[k]fluoranthene by Sphingobium sp. strain KK22 and identification of new products of non-alternant PAH biodegradation by liquid chromatography electrospray ionization tandem mass spectrometry.

    Science.gov (United States)

    Maeda, Allyn H; Nishi, Shinro; Hatada, Yuji; Ozeki, Yasuhiro; Kanaly, Robert A

    2014-03-01

    A pathway for the biotransformation of the environmental pollutant and high-molecular weight polycyclic aromatic hydrocarbon (PAH) benzo[k]fluoranthene by a soil bacterium was constructed through analyses of results from liquid chromatography negative electrospray ionization tandem mass spectrometry (LC/ESI(-)-MS/MS). Exposure of Sphingobium sp. strain KK22 to benzo[k]fluoranthene resulted in transformation to four-, three- and two-aromatic ring products. The structurally similar four- and three-ring non-alternant PAHs fluoranthene and acenaphthylene were also biotransformed by strain KK22, and LC/ESI(-)-MS/MS analyses of these products confirmed the lower biotransformation pathway proposed for benzo[k]fluoranthene. In all, seven products from benzo[k]fluoranthene and seven products from fluoranthene were revealed and included previously unreported products from both PAHs. Benzo[k]fluoranthene biotransformation proceeded through ortho-cleavage of 8,9-dihydroxy-benzo[k]fluoranthene to 8-carboxyfluoranthenyl-9-propenic acid and 9-hydroxy-fluoranthene-8-carboxylic acid, and was followed by meta-cleavage to produce 3-(2-formylacenaphthylen-1-yl)-2-hydroxy-prop-2-enoic acid. The fluoranthene pathway converged with the benzo[k]fluoranthene pathway through detection of the three-ring product, 2-formylacenaphthylene-1-carboxylic acid. Production of key downstream metabolites, 1,8-naphthalic anhydride and 1-naphthoic acid from benzo[k]fluoranthene, fluoranthene and acenaphthylene biotransformations provided evidence for a common pathway by strain KK22 for all three PAHs through acenaphthoquinone. Quantitative analysis of benzo[k]fluoranthene biotransformation by strain KK22 confirmed biodegradation. This is the first pathway proposed for the biotransformation of benzo[k]fluoranthene by a bacterium. © 2013 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  15. Analysis of co-eluted isomers of high-molecular weight polycyclic aromatic hydrocarbons in high performance liquid chromatography fractions via solid-phase nanoextraction and time-resolved Shpol'skii spectroscopy.

    Science.gov (United States)

    Wilson, Walter B; Campiglia, Andres D

    2011-09-28

    We present an accurate method for the determination of isomers of high-molecular weight polycyclic aromatic hydrocarbons co-eluted in HPLC fractions. The feasibility of this approach is demonstrated with two isomers of molecular weight 302 with identical mass fragmentation patterns, namely dibenzo[a,i]pyrene and naphtho[2,3-a]pyrene. Qualitative and quantitative analysis is carried out via laser-excited time-resolved Shpol'skii spectroscopy at liquid helium temperature. Unambiguous identification of co-eluted isomers is based on their characteristic 4.2 K line-narrowed spectra in n-octane as well as their fluorescence lifetimes. Pre-concentration of HPLC fractions prior to spectroscopic analysis is performed with the aid of gold nanoparticles via an environmentally friendly procedure. In addition to the two co-eluted isomers, the analytical figures of merit of the entire procedure were evaluated with dibenzo[a,l]pyrene, dibenzo[a,h]pyrene and dibenzo[a,e]pyrene. The analytical recoveries from drinking water samples varied between 98.2±5.5 (dibenzo[a,l]pyrene) and 102.7±3.2% (dibenzo[a,i]pyrene). The limits of detection ranged from 51.1 ng L(-1) (naphtho[2,3-a]pyrene) to 154 ng L(-1) (dibenzo[a,e]pyrene). The excellent analytical figures of merit associated to its HPLC compatibility makes this approach an attractive alternative for the analysis of co-eluted isomers with identical mass spectra. Copyright © 2011 Elsevier B.V. All rights reserved.

  16. Effects of an in-patient treatment program based on regular exercise and a balanced diet on high molecular weight adiponectin, resistin levels, and insulin resistance in adolescents with severe obesity.

    Science.gov (United States)

    Gueugnon, Carine; Mougin, Fabienne; Simon-Rigaud, Marie-Laure; Regnard, Jacques; Nègre, Véronique; Dumoulin, Gilles

    2012-08-01

    Adiponectin, the most abundant hormone produced by adipose tissue, circulates in 3 isoforms, including high molecular weight (HMW) adiponectin. The latter has been suggested to be a better predictor of metabolic disturbances and insulin resistance associated with obesity. This study investigated changes in total and HMW adiponectin, resistin, and homeostasis model assessment (HOMA) during a 9-month in-patient treatment program based on physical exercise and a balanced diet in 32 severely obese adolescents. Total and HMW adiponectin, resistin, and HOMA were measured at baseline (month 0) and during the program (months 3, 6, 9). In addition, a control group of 15 teenagers served as a reference for the baseline assessments. At baseline, HMW adiponectin was more markedly decreased in obese adolescents than total adiponectin, and both were lower than in controls. Conversely, resistin and HOMA were higher in obese adolescents. During the program, there was a significant change in body composition and improved insulin sensitivity among obese teenagers. In addition, HMW adiponectin and the ratio of HMW-to-total adiponectin increased throughout the study, whereas total adiponectin only increased up until the sixth month. On the contrary, resistin did not show any significant change. In obese adolescents, a long-term combination of aerobic exercise and a balanced diet, inducing change in body composition and improved insulin sensitivity, markedly increased HMW adiponectin compared with total adiponectin, without any change in resistin concentrations. Our results thus suggest that the determination of HMW adiponectin could be more useful than measurement of total adiponectin in clinical settings.

  17. Reduced high-molecular-weight adiponectin and elevated high-sensitivity C-reactive protein are synergistic risk factors for metabolic syndrome in a large-scale middle-aged to elderly population: the Shimanami Health Promoting Program Study.

    Science.gov (United States)

    Tabara, Yasuharu; Osawa, Haruhiko; Kawamoto, Ryuichi; Tachibana-Iimori, Rieko; Yamamoto, Miyuki; Nakura, Jun; Miki, Tetsuro; Makino, Hideich; Kohara, Katsuhiko

    2008-03-01

    In Western countries, one of the most important modifiable targets for the prevention of cardiovascular diseases is metabolic syndrome. Adiponectin is an adipose tissue-specific plasma protein that inversely associates with metabolic syndrome. Among several molecular isoforms, high-molecular-weight (HMW) complex is considered the active form. Increased serum high-sensitivity C-reactive protein (hsCRP) concentration also associates with metabolic syndrome, and adiponectin could modulate plasma C-reactive protein levels. Here, through cross-sectional investigation, we investigated whether reduced HMW adiponectin and increased hsCRP levels in plasma are synergistically associated with metabolic syndrome. Measurement of HMW complex of adiponectin is one of the novelties of this study. We analyzed 1845 community-dwelling middle-aged to elderly subjects (62+/-13 yr). Plasma HMW adiponectin levels were measured by ELISA. Clinical parameters were obtained from the subjects' personal health records, evaluated at their annual medical check-up. Each component of metabolic syndrome, except for raised blood pressure, showed significantly lower plasma HMW adiponectin concentrations for both men and women (P<0.001). In contrast, plasma hsCRP levels were significantly higher in subjects with metabolic disorders (P<0.001). After adjusting for other confounding factors, HMW adiponectin [log normalized, odds ratio 0.084 (95% confidence interval 0.050-0.142), P<0.001] and hsCRP [3.009 (2.175-4.163), P<0.001] were identified as independent determinants of metabolic syndrome. In addition to the direct associations, we also observed a synergistic effect between these two molecules (F=11.8, P<0.001). Reduced HMW adiponectin and elevated hsCRP are synergistically associated with the accumulation of metabolic disorders. The combination of these markers would be useful for identifying at-risk populations.

  18. Prokaryotic degradation of high molecular weight dissolved organic matter in the deep-sea waters of NW Mediterranean Sea under in situ temperature and pressure conditions during contrasted hydrological conditions

    Science.gov (United States)

    Tamburini, C.; Boutrif, M.; Garel, M.; Sempéré, R.; Repeta, D.; Charriere, B.; Nerini, D.; Panagiotopoulos, C.

    2016-02-01

    The contribution of the semi-labile dissolved organic carbon (DOC) to the global prokaryotic production has been assessed in very few previous studies. Some experiments show rapid utilization of semi-reactive DOC by prokaryotes, while other experiments show almost no utilization at all. However, all these studies did not take into account the role of hydrostatic pressure for the degradation of organic matter. In this study, we investigate (1) the degradation of "natural" high molecular weight DOM HMW-DOM (obtained after ultrafiltration) and (2) the uptake of labeled extracellular polymeric substances (3H-EPS) incubated with deep-sea water samples (2000 m-depth, NW Mediterranean Sea) under in situ pressure conditions (HP) and under atmospheric compression after decompression of the deep samples (ATM) during stratified and mixed water conditions (deep sea convection). Our results indicated that during HP incubations DOC exhibited the highest degradation rates (kHP DOC = 0.82 d-1) compared to the ATM conditions were no or few degradation was observed (kATM DOC= 0.007 d-1). An opposite trend was observed for the HP incubations from mixed deep water masses. HP incubation measurements displayed the lowest DOC degradation (kHP DOC=0.031 d-1) compared to the ATM conditions (kATM DOC=0.62 d-1). These results imply the presence of allochthonous prokaryotic cells in deep-sea samples after a winter water mass convection. Same trends were found using 3H-EPS uptake rates which were higher at HP than at ATM conditions during stratified period conditions whereas the opposite patterns were observed during deep-sea convection event. Moreover, we found than Euryarchaea were the main contributors to 3H-EPS assimilation at 2000m-depth, representing 58% of the total cells actively assimilating 3H-EPS. This study demonstrates that remineralization rates of semi-labile DOC in deep NW Med. Sea are controlled by the prokaryotic communities, which are influenced by the hydrological

  19. Pain relief and improved physical function in knee osteoarthritis patients receiving ongoing hylan G-F 20, a high-molecular-weight hyaluronan, versus other treatment options: data from a large real-world longitudinal cohort in Canada

    Directory of Open Access Journals (Sweden)

    Petrella RJ

    2015-10-01

    .Keywords: osteoarthritis, high-molecular-weight hyaluronic acid, intra-articular, 6-minute walk test, repeat treatment, pain relief

  20. Composition and functional analysis of low-molecular-weight glutenin alleles with Aroona near-isogenic lines of bread wheat

    Directory of Open Access Journals (Sweden)

    Zhang Xiaofei

    2012-12-01

    Full Text Available Abstract Background Low-molecular-weight glutenin subunits (LMW-GS strongly influence the bread-making quality of bread wheat. These proteins are encoded by a multi-gene family located at the Glu-A3, Glu-B3 and Glu-D3 loci on the short arms of homoeologous group 1 chromosomes, and show high allelic variation. To characterize the genetic and protein compositions of LMW-GS alleles, we investigated 16 Aroona near-isogenic lines (NILs using SDS-PAGE, 2D-PAGE and the LMW-GS gene marker system. Moreover, the composition of glutenin macro-polymers, dough properties and pan bread quality parameters were determined for functional analysis of LMW-GS alleles in the NILs. Results Using the LMW-GS gene marker system, 14–20 LMW-GS genes were identified in individual NILs. At the Glu-A3 locus, two m-type and 2–4 i-type genes were identified and their allelic variants showed high polymorphisms in length and nucleotide sequences. The Glu-A3d allele possessed three active genes, the highest number among Glu-A3 alleles. At the Glu-B3 locus, 2–3 m-type and 1–3 s-type genes were identified from individual NILs. Based on the different compositions of s-type genes, Glu-B3 alleles were divided into two groups, one containing Glu-B3a, B3b, B3f and B3g, and the other comprising Glu-B3c, B3d, B3h and B3i. Eight conserved genes were identified among Glu-D3 alleles, except for Glu-D3f. The protein products of the unique active genes in each NIL were detected using protein electrophoresis. Among Glu-3 alleles, the Glu-A3e genotype without i-type LMW-GS performed worst in almost all quality properties. Glu-B3b, B3g and B3i showed better quality parameters than the other Glu-B3 alleles, whereas the Glu-B3c allele containing s-type genes with low expression levels had an inferior effect on bread-making quality. Due to the conserved genes at Glu-D3 locus, Glu-D3 alleles showed no significant differences in effects on all quality parameters. Conclusions This work

  1. A model for the molecular structure of the glutenins from wheat flour

    NARCIS (Netherlands)

    Graveland, A.; Bosveld, P.; Lichtendonk, W.J.; Marseille, J.P.; Moonen, J.H.E.; Scheepstra, A.

    1985-01-01

    Protein fractions, which were termed glutenins I, IIa, IIb, IIIa and IIIb were isolated from flour of the Dutch wheat cultivar, Sicco, by extraction with sodium dodecyl sulphate (SDS) solution and fractionation by precipitation/redissolution techniques and by gel filtration chromatography. The

  2. Characterization and glutenin diversity in tetraploid wheat varieties ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-11-19

    Nov 19, 2008 ... Important methods applied for the breeding of bread-quality wheat (Triticum durum L.) consist of small- scale bread-quality tests for the determination of the grain protein content, SDS-sedimentation volume, thousand weight kernel and ... marked as a x and y – type subunits, based on their electrophoretic ...

  3. Segregation distortion in homozygous lines obtained via anther culture and maize doubled haploid methods in comparison to single seed descent in wheat (Triticum aestivum L.)

    OpenAIRE

    Adamski,Tadeusz; Krystkowiak,Karolina; Kuczynska,Anetta; Mikolajczak,Krzysztof; Ogrodowicz,Piotr; Ponitka,Aleksandra; Surma,Maria; Slusarkiewicz-Jarzina,Aurelia

    2014-01-01

    Background: The quality of wheat grain depends on several characteristics, among which the composition of high molecular weight glutenin subunits, encoded by Glu-1 loci, are the most important. Application of biotechnological tools to accelerate the attainment of homozygous lines may influence the proportion of segregated genotypes. The objective was to determine, whether the selection pressure generated by the methods based on in vitro cultures, may cause a loss of genotypes with desirable G...

  4. Synthesis of High-Molecular-Weight Multifunctional Glycerol Polyhydroxyurethanes PHUs

    Directory of Open Access Journals (Sweden)

    Bassam Nohra

    2016-09-01

    Full Text Available Glycerol carbonate acrylate is a 5-membered cyclic carbonate synthesized from glycerol that is used as a chemical coupling agent and has proven highly suitable for use in the synthesis of multifunctional polyhydroxyurethanes (PHUs. The multifunctionality of the structure of PHUs is determined by the density of the carbon-amine groups generated by the Aza-Michael reaction and that of the urethane groups and adjacent primary and secondary hydroxyl groups generated by aminolysis. Glycerol carbonate acrylate is polymerized with polyfunctional mono-, di-, tri, and tetra-amines, by type-AB polyaddition, either in bulk or in solution, through stepwise or one-pot reaction strategies in the absence of added catalysts. These approaches result in the generation of linear, interchain, and crosslinked structures, through the polyaddition of linear and branched amines to the ethylene and cyclic carbonate sites of glycerol carbonate acrylate. The resulting collection of organic molecules gives rise to polyethylene amino ester PHUs with a high molar mass, exceeding 20,000 g·mol−1, with uniform dispersity.

  5. High Molecular Weight Forms of Mammalian Respiratory Chain Complex II

    Czech Academy of Sciences Publication Activity Database

    Kovářová, Nikola; Mráček, Tomáš; Nůsková, Hana; Holzerová, Eliška; Vrbacký, Marek; Pecina, Petr; Hejzlarová, Kateřina; Klučková, Katarína; Rohlena, Jakub; Neužil, Jiří; Houštěk, Josef

    2013-01-01

    Roč. 8, č. 8 (2013), e71869 E-ISSN 1932-6203 R&D Projects: GA ČR(CZ) GPP303/10/P227; GA MŠk(CZ) LL1204; GA MZd(CZ) NT12370; GA ČR(CZ) GAP301/10/1937 Institutional research plan: CEZ:AV0Z50520701 Institutional support: RVO:67985823 Keywords : supercomplexes * high molecular weihgt forms of complex II * native electrophoretic systems Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.534, year: 2013

  6. Modulatory effect of high molecular weight polyethylene glycols on ...

    African Journals Online (AJOL)

    ADOWIE PERE

    (MW 4000 and 6000) on drug release from ibuprofen sustained release formulation. Different batches of .... tapped density that is the ratio of the weight of the granules and the ..... conferring less resistance to tablet fracture and abrasion.

  7. High Molecular Weight Polybenzimidazole Membranes for High Temperature PEMFC

    DEFF Research Database (Denmark)

    Yang, Jingshuai; Cleemann, Lars Nilausen; Steenberg, T.

    2014-01-01

    High temperature operation of proton exchange membrane fuel cells under ambient pressure has been achieved by using phosphoric acid doped polybenzimidazole (PBI) membranes. To optimize the membrane and fuel cells, high performance polymers were synthesized of molecular weights from 30 to 94 kDa w...

  8. Rapid isolation of high molecular weight DNA from single dried ...

    African Journals Online (AJOL)

    ANAND

    For studying genetic diversity in populations of predatory coccinellid, Cryptolaemus montrouzieri. Mulsant (Coccinellidae: Coleoptera), our attempts to isolate high quality DNA from individual adult beetle using several previously reported protocols and even modifications were quite unsuccessful as the insect size was small ...

  9. Endurance of high molecular weight carboxymethyl cellulose in corrosive environments

    Science.gov (United States)

    Murodov, M. M.; Rahmanberdiev, G. R.; Khalikov, M. M.; Egamberdiev, E. A.; Negmatova, K. C.; Saidov, M. M.; Mahmudova, N.

    2012-07-01

    Lignin obtained from the waste cooking liquor, formed after soda pulping process, is used as an inhibitor of NaCMC thermo oxidative degradation in presence of in extreme conditions during drilling oil wells. In this paper the schematic process of obtaining NaCMC by the principle of "monoapparat" on the basis of cellulose produced by non-wood cellulose materials is presented.

  10. Sampling of high molecular weight hydrocarbons with adsorbent tubes

    International Nuclear Information System (INIS)

    Stroemberg, B.

    1996-12-01

    Adsorption tubes have been used to determine the content of hydrocarbons in gas samples from small scale combustion and gasification of biomass. Compounds from benzene (mw 78) to indeno (1,2,3-cd) pyrene (mw 276) have been examined. The results show that it is possible to analyze polyaromatic hydrocarbons (PAH) with 4 aromatic rings (mw 202). Detection limits for these compounds are 3 . PAH with higher molecule weight can be identified and quantified in samples with high amounts of PAH e.g. at gasification of biomass. Sampling on adsorption tubes is extremely quick and easy. The tube is inserted in the gas of interest and the sample is sucked through the tube with a pump. Sampling times of 2-10 minutes are often sufficient. High moisture content in the gas may result in losses of the most volatile compounds, when drying. Even very low concentrations of water in the tube may cause ice formation in the cold-trap and the sample will be destroyed. The analysis is unfortunately time-consuming because the desorption oven must be cooled between every analysis. This will reduce the number of samples which can be analyzed per day. The tubes can be stored for several weeks before analysis without deterioration. 4 refs, 5 figs, 3 tabs

  11. Physicochemical characterization of irradiated high molecular weight chitosan

    International Nuclear Information System (INIS)

    Rapado, Manuel; Paredes, Mayte; Altanes, Sonia; Barrera, Gisela; Otero, Isabel; Peniche, Carlos; Gonzalez, Maykel

    2006-01-01

    Chitosan is obtained by mean of partial deacetylation of chitin. Due to the diverse biological properties of this polymer it has valuable medical applications. Some of these applications require sterile materials. For this purpose irradiation techniques seem to have advantages since they do not require any additives, which could contaminate the final sterile product with toxic residuals. The aims of the present study were to determinate the bio burden for assessing the sterilization dose and to identify the influence of the absorbed dose of gamma radiation on the molar mass and chemical structure of chitosan. Changes in polymer features were evaluated by comparing the results obtained from the characterization of treated and raw polymers. The obtained results have been shown chain cleavage caused by irradiation. It was revealed by a decrease in the intrinsic viscosities of the polymers. The invariance of the infrared spectra of polymer indicates that chain degradation occurs without significant change of the chemical structure. The results obtained have practical implication in the field of radiation sterilization of chitosan used for biotechnology, medicine and pharmacy

  12. Influence of the graphite type in the mechanical properties of graphene nanosheets and ultra high molecular weight polyethylene (GN/UHMWPE) nanocomposites; Influencia do tipo de grafite nas propriedades mecanicas do nanocomposito de nanolaminas de grafeno com polietileno de ultra alta massa molar (NG/PEUAPM)

    Energy Technology Data Exchange (ETDEWEB)

    Lima, M.E. de A.; Coutinho, S.V.; Fim, F. de C., E-mail: fabianafim@ct.ufpb.br [Universidade Federal da Paraiba (UFPB), Joao Pessoa, PB (Brazil). Departamento de Engenharia de Materiais

    2016-07-01

    This paper studies the mechanical properties of graphene nanosheets and ultra high molecular weight polyethylene (GN/UHMWPE) nanocomposites. The graphene nanosheets were obtained by two methods: the first, from Graphite Micrograf HC11 after 8 hours in a ultrasound bath in 70% alcoholic solution; the second, from Graflake 9950 prepared by oxidizing with H{sub 2}SO{sub 4}/HNO{sub 3} (4:1), followed by thermal expansion at 1050°C for 30 seconds and ultrasound bath for 20 hours. Both were oven dried for 24 hours at 110°C and subsequently added to the polymer in percentages of 0.5, 1.0 and 1.5% (w/w). The powders of the nanocomposites were mixed in a ball mill for 1 hour and molded by hot compression. XRD and SEM images was possible to verify that the Graflake's GN had higher delamination that the Micrograph's GN. The mechanical properties were affected in different percentages for each graphite type. (author)

  13. Effects of water deficit on breadmaking quality and storage protein compositions in bread wheat (Triticum aestivum L.).

    Science.gov (United States)

    Zhou, Jiaxing; Liu, Dongmiao; Deng, Xiong; Zhen, Shoumin; Wang, Zhimin; Yan, Yueming

    2018-03-12

    Water deficiency affects grain proteome dynamics and storage protein compositions, resulting in changes in gluten viscoelasticity. In this study, the effects of field water deficit on wheat breadmaking quality and grain storage proteins were investigated. Water deficiency produced a shorter grain-filling period, a decrease in grain number, grain weight and grain yield, a reduced starch granule size and increased protein content and glutenin macropolymer contents, resulting in superior dough properties and breadmaking quality. Reverse phase ultra-performance liquid chromatography analysis showed that the total gliadin and glutenin content and the accumulation of individual components were significantly increased by water deficiency. Two-dimensional gel electrophoresis detected 144 individual storage protein spots with significant accumulation changes in developing grains under water deficit. Comparative proteomic analysis revealed that water deficiency resulted in significant upregulation of 12 gliadins, 12 high-molecular-weight glutenin subunits and 46 low-molecular-weight glutenin subunits. Quantitative real-time polymerase chain reaction analysis revealed that the expression of storage protein biosynthesis-related transcription factors Dof and Spa was upregulated by water deficiency. The present results illustrated that water deficiency leads to increased accumulation of storage protein components and upregulated expression of Dof and Spa, resulting in an improvement in glutenin strength and breadmaking quality. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

  14. Moldagem por compressão a frio do polietileno de ultra alto peso molecular. Parte 1: influência do tamanho, distribuição e morfologia da partícula na densidade a verde Cold compression molding of ultra high molecular weight polyethylene. Part 1: influence of the size, distribution and morphology of particles on the green density

    Directory of Open Access Journals (Sweden)

    Bernadete A. Bittencourt

    2009-01-01

    Full Text Available Neste trabalho foram investigadas as características de amostras de pó de polietileno de ultra alto peso molecular (PEUAPM, tais como porosidade, morfologia, tamanho médio e distribuição de partícula, que são importantes na moldagem por compressão a frio. Também foi avaliada a influência dessas características na densidade a verde de pré-formas. As amostras dos pós foram caracterizadas por calorimetria diferencial de varredura (DSC, análise granulométrica, absorção de óleo, área superficial, porosimetria de mercúrio, fluidez do pó, densidade de compactação, densidade aparente e microscopia eletrônica de varredura (MEV. Através das técnicas de caracterização estudadas ficou evidenciado que as características da partícula citadas anteriormente, assim como o parâmetro de densificação (DP, que é função direta da porosidade interparticular, favorecem a densidade a verde relativa (DVR e consequentemente a tensão de resistência à flexão (TRF.In this paper an investigation was made of the characteristics of Ultra High Molecular Weight Polyethylene (UHMWPE powder samples, including porosity, particles average size, size distribution and morphology, which are important in cold compression molding. The influence of these characteristics on the green density of molded pre-shapes was also investigated. The UHMWPE powder samples were characterized by Differential Scanning Calorimetry (DSC, granulometric analysis, oil absorption, surface area, mercury porosity, density compaction, apparent density and Scanning Electron Microscopy (SEM. The characterization techniques used demonstrate that the UHMWPE particles characteristics cited above as well as the densification parameter (DP, which is a direct function of the interparticles porosity, affect the relative green density (RGD and hence, the flexural tensile strength (FTS.

  15. Wheat IgE-mediated food allergy in European patients: alpha-amylase inhibitors, lipid transfer proteins and low-molecular-weight glutenins

    DEFF Research Database (Denmark)

    Pastorello, Elide A; Farioli, Laura; Conti, Amedeo

    2007-01-01

    for sodium dodecyl sulfate-polyacrylamide gel electrophoresis/immunoblotting of the three Osborne's protein fractions (albumin/globulin, gliadins and glutenins) of raw and cooked wheat. Thermal sensitivity of wheat lipid transfer protein (LTP) was investigated by spectroscopic approaches. IgE cross...

  16. New insight into the solution structures of wheat gluten proteins from Raman optical activity

    DEFF Research Database (Denmark)

    Blanch, E.W.; Kasarda, D.D.; Hecht, L.

    2003-01-01

    Vibrational Raman optical activity (ROA) spectra of the wheat proteins a-gliadin (A-gliadin), omega-liadin, and a 30 kDa peptide called T-A-1 from the high molecular weight glutenin subunit (HMW-GS) Dx5 were measured to obtain new information about their solution structures. The spectral data show...... that, under the conditions investigated, A-gliadin contains a considerable amount of hydrated alpha-helix, most of which probably lies within a relatively structured C-terminal domain. Smaller quantities of beta-structure and poly(L-proline) II (PPII) helix were also identified. Addition of methanol...

  17. Sodium Dodecyl Sulfate- Polyacrylamide Gel Electrophoresis (SDS- PAGE) of Irradiated Wheat Flour Proteins

    International Nuclear Information System (INIS)

    Souzan, R.M.

    1999-01-01

    Sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) of wheat (Triticum aestivum L) flour have revealed 23 polypeptides of molecular weights between 170 and 11.57 KDa, High molecular weight glutenin subunits (LMW-GS) were distinguished. Densitometric analysis of the gel showed the effect of radiation on polypeptide constitution at radiation energy up to 7.5 kGy. Irradiation of wheat flour with 2.5 kGy have resulted in a slight increase in the molecular weight of wheat flour protein subunits. The increase of irradiation dose to 5.0 kGy has also induced an additional increase of molecular weight of protein subunits. The continuity in application of more radiation energy to a level of 7.5 kGy have resulted in the prevalence of degradation processes of all protein subunits more than the aggregation

  18. Thermal behavior of native and hydrophobized wheat gluten, gliadin and glutenin-rich fractions by modulated DSC.

    Science.gov (United States)

    Micard, V; Guilbert, S

    2000-06-13

    The glass transition temperature (T(g)) of hydrophobized and native wheat gluten and its protein fractions, with water mass fraction from 0 to 0.2, was studied using modulated differential scanning calorimetry. The T(g) values of unplasticized products were approximately 175 degrees C whatever the treatment (hydrophobization) or the fraction tested, except for the gliadin-rich fraction (162 degrees C). Experimental change in heat capacity at the glass transition (DeltaC(p)) ranged from 0.32 to 0. 50 J/g/ degrees C depending on the gluten fractions. The Gordon-Taylor fit of T(g) evolution as a function of water content showed that glutenin-rich fractions were more sensitive to water plasticization than the gliadin-rich fraction. The Kwei equation gave better fit to experimental data and demonstrated that the water plasticization of gluten and its fractions is influenced by secondary interactions. However, the application of the Couchman-Karasz equation without fitting predicts satisfactorily the plasticization of gluten proteins by water.

  19. Highly conserved small subunit residues influence rubisco large subunit catalysis.

    Science.gov (United States)

    Genkov, Todor; Spreitzer, Robert J

    2009-10-30

    The chloroplast enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes the rate-limiting step of photosynthetic CO(2) fixation. With a deeper understanding of its structure-function relationships and competitive inhibition by O(2), it may be possible to engineer an increase in agricultural productivity and renewable energy. The chloroplast-encoded large subunits form the active site, but the nuclear-encoded small subunits can also influence catalytic efficiency and CO(2)/O(2) specificity. To further define the role of the small subunit in Rubisco function, the 10 most conserved residues in all small subunits were substituted with alanine by transformation of a Chlamydomonas reinhardtii mutant that lacks the small subunit gene family. All the mutant strains were able to grow photosynthetically, indicating that none of the residues is essential for function. Three of the substitutions have little or no effect (S16A, P19A, and E92A), one primarily affects holoenzyme stability (L18A), and the remainder affect catalysis with or without some level of associated structural instability (Y32A, E43A, W73A, L78A, P79A, and F81A). Y32A and E43A cause decreases in CO(2)/O(2) specificity. Based on the x-ray crystal structure of Chlamydomonas Rubisco, all but one (Glu-92) of the conserved residues are in contact with large subunits and cluster near the amino- or carboxyl-terminal ends of large subunit alpha-helix 8, which is a structural element of the alpha/beta-barrel active site. Small subunit residues Glu-43 and Trp-73 identify a possible structural connection between active site alpha-helix 8 and the highly variable small subunit loop between beta-strands A and B, which can also influence Rubisco CO(2)/O(2) specificity.

  20. Identification of Low Molecular Weight Glutenin Alleles by Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF-MS) in Common Wheat (Triticum aestivum L.)

    Science.gov (United States)

    Islam, Shahidul; Applebee, Marie; Appels, Rudi; Yan, Yueming; Ma, Wujun

    2015-01-01

    Low molecular weight glutenin subunits (LMW-GS) play an important role in determining dough properties and breadmaking quality. However, resolution of the currently used methodologies for analyzing LMW-GS is rather low which prevents an efficient use of genetic variations associated with these alleles in wheat breeding. The aim of the current study is to evaluate and develop a rapid, simple, and accurate method to differentiate LMW-GS alleles using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. A set of standard single LMW-GS allele lines as well as a suite of well documented wheat cultivars were collected from France, CIMMYT, and Canada. Method development and optimization were focused on protein extraction procedures and MALDI-TOF instrument settings to generate reproducible diagnostic spectrum peak profiles for each of the known wheat LMW-GS allele. Results revealed a total of 48 unique allele combinations among the studied genotypes. Characteristic MALDI-TOF peak patterns were obtained for 17 common LMW-GS alleles, including 5 (b, a or c, d, e, f), 7 (a, b, c, d or i, f, g, h) and 5 (a, b, c, d, f) patterns or alleles for the Glu-A3, Glu-B3, and Glu-D3 loci, respectively. In addition, some reproducible MALDI-TOF peak patterns were also obtained that did not match with any known alleles. The results demonstrated a high resolution and throughput nature of MALDI-TOF technology in analyzing LMW-GS alleles, which is suitable for application in wheat breeding programs in processing a large number of wheat lines with high accuracy in limited time. It also suggested that the variation of LMW-GS alleles is more abundant than what has been defined by the current nomenclature system that is mainly based on SDS-PAGE system. The MALDI-TOF technology is useful to differentiate these variations. An international joint effort may be needed to assign allele symbols to these newly identified alleles and determine their effects on end

  1. Overexpression of avenin-like b proteins in bread wheat (Triticum aestivum L.) improves dough mixing properties by their incorporation into glutenin polymers.

    Science.gov (United States)

    Ma, Fengyun; Li, Miao; Li, Tingting; Liu, Wei; Liu, Yunyi; Li, Yin; Hu, Wei; Zheng, Qian; Wang, Yaqiong; Li, Kexiu; Chang, Junli; Chen, Mingjie; Yang, Guangxiao; Wang, Yuesheng; He, Guangyuan

    2013-01-01

    Avenin-like b proteins are a small family of wheat storage proteins, each containing 18 or 19 cysteine residues. The role of these proteins, with high numbers of cysteine residues, in determining the functional properties of wheat flour is unclear. In the present study, two transgenic lines of the bread wheat overexpressing avenin-like b gene were generated to investigate the effects of Avenin-like b proteins on dough mixing properties. Sodium dodecyl sulfate sedimentation (SDSS) test and Mixograph analysis of these lines demonstrated that overexpression of Avenin-like b proteins in both transgenic wheat lines significantly increased SDSS volume and improved dough elasticity, mixing tolerance and resistance to extension. These changes were associated with the increased proportion of polymeric proteins due to the incorporation of overexpressed Avenin-like b proteins into the glutenin polymers. The results of this study were critical to confirm the hypothesis that Avenin-like b proteins could be integrated into glutenin polymers by inter-chain disulphide bonds, which could help understand the mechanism behind strengthening wheat dough strength.

  2. Overexpression of avenin-like b proteins in bread wheat (Triticum aestivum L. improves dough mixing properties by their incorporation into glutenin polymers.

    Directory of Open Access Journals (Sweden)

    Fengyun Ma

    Full Text Available Avenin-like b proteins are a small family of wheat storage proteins, each containing 18 or 19 cysteine residues. The role of these proteins, with high numbers of cysteine residues, in determining the functional properties of wheat flour is unclear. In the present study, two transgenic lines of the bread wheat overexpressing avenin-like b gene were generated to investigate the effects of Avenin-like b proteins on dough mixing properties. Sodium dodecyl sulfate sedimentation (SDSS test and Mixograph analysis of these lines demonstrated that overexpression of Avenin-like b proteins in both transgenic wheat lines significantly increased SDSS volume and improved dough elasticity, mixing tolerance and resistance to extension. These changes were associated with the increased proportion of polymeric proteins due to the incorporation of overexpressed Avenin-like b proteins into the glutenin polymers. The results of this study were critical to confirm the hypothesis that Avenin-like b proteins could be integrated into glutenin polymers by inter-chain disulphide bonds, which could help understand the mechanism behind strengthening wheat dough strength.

  3. Characterization of fimbrial subunits from Bordetella species

    NARCIS (Netherlands)

    Mooi, F.R.; Heide, H.G.J. van der; Avest, A.R. ter; Welinder, K.G.; Livey, I.; Zeijst, B.A.M. van der; Gaastra, W.

    Using antisera raised against serotype 2 and 3 fimbrial subunits from Bordetella pertussis, serologically related polypeptides were detected in Bordetella bronchiseptica, Bordetella parapertussis and Bordetella avium strains. The two B. pertussis fimbrial subunits, and three of the serologically

  4. Neutronic performance of high molecular weight coolants for a prismatic VHTR

    International Nuclear Information System (INIS)

    Schriener, T. M.; El-Genk, M. S.

    2008-01-01

    A neutronic model is developed of a prismatic Very High Temperature Reactor (VHTR) to investigate the effects on the excess reactivity and operation cycle length of replacing helium with binary gas mixtures of He-Ne, He-N 2 , or He-Xe as reactor coolants and working fluids in the direct Closed Brayton Cycle (CBC) for energy conversion. Also investigated is the neutron activation of these binary gas mixtures in the VHTR. The motivation for using the heavy binary mixtures is the smaller size and the fewer number of stages of the CBC turbo-machinery. The present analysis uses the Monte Carlo code MCNPX 2.6D at typical operating conditions (500-1000 degrees and 7.12 MPa) in the VHTR. He-Ne (15 g/mol) is the best neutronically, but not thermal-hydraulically, followed by He-N 2 . Although He-Ne has ∼13.6% lower heat transfer coefficient than helium, it insignificantly affects the initial excess reactivity and the operation life cycle and experiences no neutrons activation. On the other hand, He-N 2 has 4.4% higher heat transfer coefficient than helium and experiences insignificant neutron activation in the reactor, but decreases the initial excess reactivity by ∼5.2% and the operation cycle length by 6.7%. He-Xe (15 g/mol) has 8% higher heat transfer coefficient than helium, but decreases the initial excess reactivity by 18.2% and the operational cycle length by 17%. In addition, neutron activation of xenon produces a significant source term, requiring shielding of the CBC loop and could contaminate the turbo-machinery with long-lived radioactive cesium. Thus, He-Xe is not recommended as a reactor coolant, but could be used as working fluid in a CBC loop that is indirectly coupled to helium cooled VHTR. (authors)

  5. Investigation of Plasma Eects in Ultra High Molecular Weight Polyethylene (UHMWPE) Cords

    DEFF Research Database (Denmark)

    Teodoru, Steluta; Kusano, Yukihiro; Rozlosnik, Noemi

    modication for improved wetting and/or adhesion with other polymeric materials. Atmospheric pressure plasma treatment is promising for this purpose due to its environmental compatibility, high treatment eects without aecting the textural characteristics of the bulk material, its applicability to a variety...... of shapes, and easy up-scaling and construction of in-line production processes. An atmospheric pressure dielectric barrier discharge (DBD) plasma is used to study surface modication eect on UHMWPE cords, operated at a frequency of ca. 40 kHz in He, He/O2, O2 and N2 gases. The cords were continuously...

  6. Continuous Plasma Treatment of Ultra-High-Molecular-Weight Polyethylene (UHMWPE) Fibres for Adhesion Improvement

    DEFF Research Database (Denmark)

    Teodoru, Steluta; Kusano, Yukihiro; Rozlosnik, Noemi

    2009-01-01

    A dielectric barrier discharge in Ar, He, He/O2, N2 or O2 at atmospheric pressure was used for the continuous plasma treatment of UHMWPE fibres. The influence of the input power of the discharge and the gas flow rate on surface modification is studied with the aim of adhesion improvement. Surface...

  7. High-molecular-weight HPMA-based polymer drug carriers for delivery to tumor

    Czech Academy of Sciences Publication Activity Database

    Kostka, Libor; Etrych, Tomáš

    2016-01-01

    Roč. 65, Suppl. 2 (2016), S179-S190 ISSN 0862-8408 R&D Projects: GA MŠk(CZ) LO1507 Institutional support: RVO:61389013 Keywords : nanotherapeutics * pH responsive * HPMA copolymers Subject RIV: CD - Macromolecular Chemistry Impact factor: 1.461, year: 2016 http://www.biomed.cas.cz/physiolres/pdf/65%20Suppl%202/65_S179.pdf

  8. High molecular weight polyurethanes and a polyurethane urea based on 1,4-butanediisocyanate

    NARCIS (Netherlands)

    Spaans, CJ; de Groot, JH; Dekens, FG; Pennings, AJ

    New biomedical polyurethanes and a polyurethane urea based on epsilon-caprolactone and 1,4-butanediisocyanate have been developed. On degradation, only non-toxic products are produced. The polyurethane urea with poly(epsilon-caprolactone) soft segments and butanediisocyanate/butanediamine hard

  9. Studies on a microbially derived, high molecular weight inhibitor of plasma cholesteryl ester transfer protein

    International Nuclear Information System (INIS)

    Marschke, C.K.; McGee, J.E.; Melchior, G.W.; Castle, C.K.

    1989-01-01

    The authors have isolated an organism which accumulates an inhibitor of Cholesteryl Ester Transfer Protein (CETP). Purification of 100,000-fold was achieved by ammonium sulfate precipitation followed by Hydroxyl Apatite, Agarose AO.5, and Mono Q (Pharmacia) chromatographies. The use of 14 C-labelled protein molecular weight standards followed by SDS-PAGE revealed some proteolytic activity. However, inhibition of the proteases did not affect the inhibitor potency. The inhibitor has an estimated molecular weight of 40 Kd and appears to exist as two forms. One form was eluted from a Mono Q column by 100 mM NaCl while the other was not bound. Our evidence indicated that the bound form was progressively denatured, or proteolyzed, during storage of the fermentation beer, to the unbound form. Importantly though this molecular change did not affect either inhibitory activity or the apparent molecular weight

  10. Characterization and quantitation of low and high molecular weight phenolic compounds in apple seeds.

    Science.gov (United States)

    Fromm, Matthias; Bayha, Sandra; Carle, Reinhold; Kammerer, Dietmar R

    2012-02-08

    The phenolic constituents of seeds of 12 different apple cultivars were fractionated by sequential extraction with aqueous acetone (30:70, v/v) and ethyl acetate after hexane extraction of the lipids. Low molecular weight phenolic compounds were individually quantitated by RP-HPLC-DAD. The contents of extractable and nonextractable procyanidins were determined by applying RP-HPLC following thiolysis and n-butanol/HCl hydrolysis, respectively. As expected, the results revealed marked differences of the ethyl acetate extracts, aqueous acetone extracts, and insoluble residues with regard to contents and mean degrees of polymerization of procyanidins. Total phenolic contents in the defatted apple seed residues ranged between 18.4 and 99.8 mg/g. Phloridzin was the most abundant phenolic compound, representing 79-92% of monomeric polyphenols. Yields of phenolic compounds significantly differed among the cultivars under study, with seeds of cider apples generally being richer in phloridzin and catechins than seeds of dessert apple cultivars. This is the first study presenting comprehensive data on the contents of phenolic compounds in apple seeds comprising extractable and nonextractable procyanidins. Furthermore, the present work points out a strategy for the sustainable and complete exploitation of apple seeds as valuable agro-industrial byproducts, in particular as a rich source of phloridzin and antioxidant flavanols.

  11. Ballistic behaviour of ultra-high molecular weight polyethylene: effect of gamma radiation

    International Nuclear Information System (INIS)

    Alves, Andreia L.S.; Nascimento, Lucio F.C.; Miguez Suarez, Joao Carlos

    2004-01-01

    The fiber reinforced polymer matrix composites (PMCs) are considered excellent engineering materials. In structural applications, when a high strength-to-weight ratio is fundamental for the design, PMCs are successfully replacing many conventional materials. Since World War II textile materials have been used as ballistic armor. Materials manufactured with ultrahigh molecular weight polyethylene (UHMWPE) fibers are used in the production of armor materials, for personnel protection and armored vehicles. As these have been developed and commercialized more recently, there is not enough information about the action of the ionizing radiation in the ballistic performance of this armor material. In the present work the ballistic behavior of composite plates manufactured with ultrahigh molecular weight polyethylene (UHMWPE) fibers were evaluated after exposure to gamma radiation. The ballistic tests results were related to the macromolecular modifications induced by the environmental degradation through mechanical (hardness, impact and flexure) and physicochemical (infrared spectroscopy, differential scanning calorimetry and thermal gravimetric analysis) tests. Our results indicate that gamma irradiation induces modifications in the UHMWPE macromolecular chains, altering the mechanical properties of the composite and decreasing, for higher radiation doses, its ballistic performance. These results are presented and discussed. (author)

  12. Hydrolytically and reductively degradable high-molecular-weight poly(ethylene glycol)s

    Czech Academy of Sciences Publication Activity Database

    Braunová, Alena; Pechar, Michal; Laga, Richard; Ulbrich, Karel

    2007-01-01

    Roč. 208, č. 24 (2007), s. 2642-2653 ISSN 1022-1352 R&D Projects: GA AV ČR KAN200200651; GA MŠk 1M0505 Institutional research plan: CEZ:AV0Z40500505 Keywords : biodegradable * drug delivery systems * gene delivery vectors * poly(ethylene glycol) Subject RIV: CE - Biochemistry Impact factor: 2.046, year: 2007

  13. A Simple Thermoplastic Substrate Containing Hierarchical Silica Lamellae for High-Molecular-Weight DNA Extraction.

    Science.gov (United States)

    Zhang, Ye; Zhang, Yi; Burke, Jeffrey M; Gleitsman, Kristin; Friedrich, Sarah M; Liu, Kelvin J; Wang, Tza-Huei

    2016-12-01

    An inexpensive, magnetic thermoplastic nanomaterial is developed utilizing a hierarchical layering of micro- and nanoscale silica lamellae to create a high-surface-area and low-shear substrate capable of capturing vast amounts of ultrahigh-molecular-weight DNA. Extraction is performed via a simple 45 min process and is capable of achieving binding capacities up to 1 000 000 times greater than silica microparticles. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Appearance and partial purification of a high molecular weight protein in crabs exposed to saxitoxin.

    Science.gov (United States)

    Barber, K G; Kitts, D D; Townsley, P M; Smith, D S

    1988-01-01

    This paper provides evidence for a protein component which appears to be involved in the seasonal resistance of small shore crabs, Hemigrapsus oregonesis and Hemigrapsus nudus to saxitoxin, a principle neurotoxin involved in paralytic shellfish poisoning (PSP). This unique protein complex was isolated and partially purified by ion exchange chromatography using DEAE-cellulose from visceral tissue extracts of resistant crabs. The complex was absent in control crabs that were sensitive to saxitoxin. In addition, the protein complex was induced in the crab after acute administration of low doses of saxitoxin. Results indicate that the protein complex is acidic in nature and has an apparent mol. wt of 145,000.

  15. Identification of components in High Molecular Weight Products: Their properties and mechanism of formation

    DEFF Research Database (Denmark)

    Hjorth, Christian Fogt

    2015-01-01

    of insulin HMWP is needed. This thesis comprises identification of the primary structure of human insulin HMWP and its biophysical and structural properties. 16 species of HMWP were identified in formulations of human insulin containing zinc and m-cresol. All identified species were covalent dimers...... investigations of insulin self-association in zinc-free environments. The effect on insulin fibrillation could also be investigated further for potentially uncoveing novel mechanisms for prevention of fibrillation in protein formulations....

  16. Identification of high molecular weight nitroaromatic compounds from coal fly ash

    International Nuclear Information System (INIS)

    Harris, W.R.; Okamoto, D.J.; Chess, E.K.; Wilson, B.W.

    1983-01-01

    A large sample of stack-collected coal fly ash was extracted with 60:40 nu/nu benzene:methanol to remove as much of the soluble organic material as possible. This solution was concentrated by gentle evaporation, and was then fractionated on a series of high performance liquid chromatography columns to generate samples suitable for probe mass spectrometric analysis. A series of nitrated derivatives of C 21 H 12 polycyclic aromatic hydrocarbon have been tentatively identified by low and high resolution mass spectrometry and gas chromatography. The series includes a mononitro, two dinitro isomers, and a trinitro derivative

  17. Immunoblotting studies of coagulation factor XII, plasma prekallikrein, and high molecular weight kininogen

    International Nuclear Information System (INIS)

    Laemmle, B.B.; Berrettini, M.; Griffin, J.H.

    1987-01-01

    Immunoblotting techniques for the qualitative and quantitative analysis of FXII, PK, and HMWK in whole plasma are presented. Sensitive, specific, and quantitative immunodetection of FXII and PK can be achieved by developing the blots with polyclonal antiserum followed by radiolabeled FXII or PK, respectively. This approach is based on the assumption that bivalent antibodies bind monovalently to the NC-bound antigen and have available binding sites to bind radiolabeled antigen derived from the fluid phase. This radiolabeled antigen overlay principle may be generally useful for immunodetection of any trace protein in complex mixtures, provided that the radiolabeled purified antigen is available. Immunoblotting may also be helpful for the partial characterization of the structural or functional abnormalities of CRM-positive variant molecules. For example, earlier studies of a FXII-variant molecule that had been purified and characterized were supported by immunoblotting studies of the CRM-positive deficient plasma. Quantitative measurement of HMWK is possible using a monoclonal antibody directed against the light chain of HMWK followed by radiolabeled secondary antibody. Quantitation of cleaved and single-chain HMWK is possible using dilutions of dextran sulfate-activated NHP on unreduced SDS-PAGE and dilutions of unactivated NHP with reduced SDS-PAGE as standards. These assays allow assessment of the degree of in vivo activation of the contact system in various disease states

  18. Systemic administration of high-molecular weight hyaluronan stimulates wound healing in genetically diabetic mice.

    Science.gov (United States)

    Galeano, Mariarosaria; Polito, Francesca; Bitto, Alessandra; Irrera, Natasha; Campo, Giuseppe M; Avenoso, Angela; Calò, Margherita; Lo Cascio, Patrizia; Minutoli, Letteria; Barone, Mauro; Squadrito, Francesco; Altavilla, Domenica

    2011-07-01

    Hyaluronic acid (HA), an essential component of the extracellular matrix, is an efficient space filler that maintains hydration, serves as a substrate for assembly of proteoglycans and is involved in wound healing. Although numerous pieces of evidence demonstrate beneficial effects in promoting wound healing in diabetes, a systemic approach has never been tested. We used an incisional wound healing model in genetically diabetic mice to test the effects of systemic injection of HA. Diabetic (n=56) and normoglycemic (n=56) mice were subjected to incision and randomized (8 groups of 7 animals each) to receive HA at different doses, 7.5, 15 and 30mg/kg/i.p., or vehicle (0.9% NaCl solution) for 12days. At the end of the experiment animals were sacrificed and skin wounds were excised for histological, biochemical and molecular analysis. Histology revealed that the most effective dose to improve wound repair and angiogenesis in diabetic mice was 30mg/kg. Furthermore HA injection (30mg/kg) improved the altered healing pattern in diabetic animals, increased skin remodeling proteins TGF-β and transglutaminase-II and restored the altered expression of cyclin B1/Cdc2 complex. Evaluation of skin from diabetic animals injected with HA revealed also an increase in HA content, suggesting that systemic injection may be able to restore the reduced intracellular HA pool of diabetic mice. Finally HA markedly improved skin mechanical properties. These promising results, if confirmed in a clinical setting, may improve the care and management of diabetic patients. Copyright © 2011 Elsevier B.V. All rights reserved.

  19. Method for Making High Molecular Weight, Extended pi-Conjugated Polymers

    Science.gov (United States)

    2001-05-04

    derivatized poly(terephthalates)s as coatings for electronics components, and as construction materials for field- effect transistors, both applications...mannose, dulose, idose, galactose and talose; ketoses such as erythrulose, ribulose, xylulose, psicose, fructose, sorbose, tagatose ; di-, tri-, 12...show that Sc(OTr)3 alone was not effective as a 15 polymerization catalyst. When the organic salt was introduced, Sc(OTr)3 became marginally

  20. Purification of high molecular weight genomic DNA from powdery mildew for long-read sequencing

    Science.gov (United States)

    The powdery mildew fungi are a group of economically important fungal plant pathogens. Relatively little is known about the molecular biology and genetics of these pathogens, in part due to a lack of well-developed genetic and genomic resources. These organisms have large, repetitive genomes, which ...

  1. Modifications by γ irradiation of polyethylene at very high molecular weight. Tribological behaviour for orthopedic applications

    International Nuclear Information System (INIS)

    Chaix, N.

    2001-07-01

    A new mode of sterilization by vacuum gamma irradiation is studied. This new mode improves significantly the polyethylene behaviour during the study in friction-corrosion against a stainless steel-316L sphere. (N.C.)

  2. High molecular weight lectin isolated from the mucus of the giant African snail Achatina fulica.

    Science.gov (United States)

    Ito, Shigeru; Shimizu, Masahiro; Nagatsuka, Maki; Kitajima, Seiji; Honda, Michiyo; Tsuchiya, Takahide; Kanzawa, Nobuyuki

    2011-01-01

    To understand better the host defense mechanisms of mollusks against pathogens, we examined the anti-microbial activity of mucus from the giant African snail Achatina fulica. Hemagglutination activity of the mucus secreted by the integument of snails inoculated with Escherichia coli was observed to increase and to cause hemagglutination of rabbit red blood cells. Purification of the snail mucus lectin by sequential column chromatography revealed that the relative molecular mass of the lectin was 350 kDa. The hemagglutination activity of the lectin was Ca(2+)-dependent and was inhibited by galactose. Growth arrest tests showed that the lectin did not inhibit bacterial growth, but did induce agglutination of gram-positive and gram-negative bacteria. Tissue distribution analyses using a polyclonal antibody revealed that the lectin was expressed in the tissues of the mantle collar. The lectin isolated from the mucus of the snail appeared to contribute to its innate immunity.

  3. Genomics of high molecular weight plasmids isolated from an on-farm biopurification system

    NARCIS (Netherlands)

    Martini, Maria C.; Wibberg, Daniel; Lozano, Mauricio; Torres Tejerizo, Gonzalo; Albicoro, Francisco J.; Jaenicke, Sebastian; van Elsas, Jan Dirk; Petroni, Alejandro; Pilar Garcillan-Barcia, M.; de la Cruz, Fernando; Schlueter, Andreas; Puehler, Alfred; Pistorio, Mariano; Lagares, Antonio; Del Papa, Maria F.

    2016-01-01

    The use of biopurification systems (BPS) constitutes an efficient strategy to eliminate pesticides from polluted wastewaters from farm activities. BPS environments contain a high microbial density and diversity facilitating the exchange of information among bacteria, mediated by mobile genetic

  4. Modeling Ballistic Response of Ultra-High-Molecular-Weight Polyethylene (UHMWPE)

    Science.gov (United States)

    2016-07-01

    in high-speed penetration problems , the material would fail and erode, and the regular contact algorithm cannot update the contact surfaces...High velocity impact and armour design. Express Polymer Letters. 2011;5(3):262–272. 14. Chocron S, King N, Walker JD, Heisserer U, Werff H

  5. High Performance Shape Memory Polyurethane Synthesized with High Molecular Weight Polyol as the Soft Segment

    Directory of Open Access Journals (Sweden)

    Manzoor Ahmad

    2012-05-01

    Full Text Available Shape memory polyurethanes (SMPUs are typically synthesized using polyols of low molecular weight (MW~2,000 g/mol as it is believed that the high density of cross-links in these low molecular weight polyols are essential for high mechanical strength and good shape memory effect. In this study, polyethylene glycol (PEG-6000 with MW ~6000 g/mol as the soft segment and diisocyanate as the hard segment were used to synthesize SMPUs, and the results were compared with the SMPUs with polycaprolactone PCL-2000. The study revealed that although the PEG-6000-based SMPUs have lower maximum elongations at break (425% and recovery stresses than those of PCL-based SMPUs, they have much better recovery ratios (up to 98% and shape fixity (up to 95%, hence better shape memory effect. Furthermore, PEG-based SMPUs showed a much shorter actuation time of < 10 s for up to 90% shape recovery compared to typical actuation times of tens of seconds to a few minutes for common SMPUs, demonstrated their great potential for applications in microsystems and other engineering components.

  6. Surface modification of ultra-high molecular weight polyethylene (UHMWPE) by argon plasma

    International Nuclear Information System (INIS)

    Liu Hengjun; Pei Yanan; Xie Dong; Deng Xingrui; Leng, Y.X.; Jin Yong; Huang Nan

    2010-01-01

    In this work, argon (Ar) plasma generated by microwave electron cyclotron resonance (MWECR) has been used to modify the UHMWPE in order to increase the wear resistance. The results showed that the wettability, anti-scratch and wear resistance of UHMWPE treated by the Ar plasma had been improved, comparing with native UHMWPE. The FTIR and XPS spectra indicated the improvement of wettability should come from the oxygen based functional groups generated on the surface of UHMWPE. The improvement of anti-scratch and wear resistance may come from the enhancement of crosslinking of UHMWPE by Ar plasma treatment.

  7. Combustion of High Molecular Weight Hydrocarbon Fuels and JP-8 at Moderate Pressures

    Science.gov (United States)

    2016-07-26

    1. Introduction Fundamental knowledge of mechanisms of autoignition of condensed hydrocarbon fuels at elevated pressures is essential for accurate...particular JP-8) and surrogates of jet-fuels in laminar non-uniform flows at elevated pressures upto 2.5 MPa. Experimental and kinetic modeling studies...AGENCY NAME(S) AND ADDRESS (ES) U.S. Army Research Office P.O. Box 12211 Research Triangle Park, NC 27709-2211 Combustion, Jet Fuels, JP-8, Elevated

  8. The physical and chemical properties of plasma treated ultra-high-molecular-weight polyethylene fibers

    DEFF Research Database (Denmark)

    Kusano, Yukihiro; Teodoru, Steluta; Hansen, Charles M.

    2011-01-01

    A uniform and smooth transfer of stresses across the polymer matrix/fiber interface is enhanced when adhesion between the matrix and fiber surface is optimized. In the absence of covalent bonds matching the Hansen solubility (cohesion) parameters (HSP) of the fiber surface with the HSP of a matrix...... at 18.0, 1.2, and 1.4, all in MPa½. The dispersion parameter has been lowered somewhat by the plasma treatment, while the polar and hydrogen bonding parameters are much higher. The HSP methodology predicts enhanced adhesion is possible by skillful use of anhydride and nitrile functional groups in matrix...

  9. Bioaccessible Porosity in Soil Aggregates and Implications for Biodegradation of High Molecular Weight Petroleum Compounds.

    Science.gov (United States)

    Akbari, Ali; Ghoshal, Subhasis

    2015-12-15

    We evaluated the role of soil aggregate pore size on biodegradation of essentially insoluble petroleum hydrocarbons that are biodegraded primarily at the oil-water interface. The size and spatial distribution of pores in aggregates sampled from biodegradation experiments of a clayey, aggregated, hydrocarbon-contaminated soil with relatively high bioremediation end point were characterized by image analyses of X-ray micro-CT scans and N2 adsorption. To determine the bioaccessible pore sizes, we performed separate experiments to assess the ability of hydrocarbon degrading bacteria isolated from the soil to pass through membranes with specific sized pores and to access hexadecane (model insoluble hydrocarbon). Hexadecane biodegradation occurred only when pores were 5 μm or larger, and did not occur when pores were 3 μm and smaller. In clayey aggregates, ∼ 25% of the aggregate volume was attributed to pores larger than 4 μm, which was comparable to that in aggregates from a sandy, hydrocarbon-contaminated soil (~23%) scanned for comparison. The ratio of volumes of inaccessible pores (4 μm) in the clayey aggregates was 0.32, whereas in the sandy aggregates it was approximately 10 times lower. The role of soil microstructure on attainable bioremediation end points could be qualitatively assessed in various soils by the aggregate characterization approach outlined herein.

  10. High molecular weight hyaluronan mediates the cancer resistance of the naked mole-rat

    OpenAIRE

    Tian, Xiao; Azpurua, Jorge; Hine, Christopher; Vaidya, Amita; Myakishev-Rempel, Max; Ablaeva, Julia; Mao, Zhiyong; Nevo, Eviatar; Gorbunova, Vera; Seluanov, Andrei

    2013-01-01

    The naked mole-rat displays exceptional longevity, with a maximum lifespan exceeding 30 years 1–3 . This is the longest reported lifespan for a rodent species and is especially striking considering the small body mass of the naked mole-rat. In comparison, a similarly sized house mouse has a maximum lifespan of 4 years 4,5 . In addition to their longevity, naked mole-rats show an unusual resistance to cancer. Multi-year observations of large naked mole-rat colonies did not detect a single inci...

  11. Synthesis and characterization of high molecular weight hydrophobically modified polyacrylamide nanolatexes using novel nonionic polymerizable surfactants

    Directory of Open Access Journals (Sweden)

    A.M. Al-Sabagh

    2013-12-01

    Full Text Available In this article, nine hydrophobically modified polyacrylamides (HM-PAM nanolatexes, were synthesized by copolymerizing the acrylamide monomer and novel polymerizable surfactants (surfmers. The reaction was carried out by inverse microemulsion copolymerization technique. The copolymerization was initiated by redox initiators composed of potassium peroxodisulphate and sodium bisulfite. The emulsion was stabilized using mixed tween 85 and span 80 as nonionic emulsifiers. The prepared HM-PAMs were classified into three groups according to the surfmers used in the copolymerization. The chemical structures of the prepared HM-PAMs were confirmed by FT-IR, 1H NMR and 13C NMR. The thermal properties were estimated with the thermal gravimetric analysis (TGA. The size and morphology of the prepared latexes were investigated by the dynamic light scattering (DLS and the High Resolution Transmission Electron Microscope (HRTEM. Finally, the molecular weights of the prepared copolymers were determined by the GPC and the viscosity average molecular weight method. They were situated between 1.58 × 106 and 0.89 × 106.

  12. Optimization of high molecular weight pullulan production by Aureobasidium pullulans in batch fermentations.

    Science.gov (United States)

    Gibson, Larry H; Coughlin, Robert W

    2002-01-01

    Of five strains of Aureobasidium pullulans studied, NRRL Y-2311-1 yielded the highest titer (26.2 g/L) of pullulan and formed the lowest amount of melanin-like pigment. Sucrose was superior to glucose as the carbon and energy source on the basis of yield and titer of pullulan produced. Pullulan titer was higher (26.2 vs 5.1 g/L), biomass concentration was lower (6.9 vs 12.7 g/L), and DO was lower (0 vs 60% of saturation) when the fermenter was agitated by a marine propeller compared to Rushton impellers. Pullulan produced by strain NRRL Y-2311-1 ranged in weight-average molar mass (M(w)) from 486 KDa and number-average molar mass (M(n)) from 220 Da on day 1 of growth to 390 KDa and 690 Da on day 6; M(w) declined by about 35% from day 1 to day 3, the day of maximum pullulan titer. For the other strains, the ranges of molar mass on the day of maximum pullulan titer were 338-614 KDa (M(w)) and 100-6820 Da (M(n)).

  13. Development of Thin-film Dye-sensitized Photoactive Materials on Ultra High Molecular Weight Polyethylene

    Science.gov (United States)

    2012-04-01

    although it is used as a base in most paints due to its light scattering ability. In general, a pure anatase TiO2 nanostructure is preferred over...solution, 0.1%, 0.5%, and 1% of pristine graphene was added to each solution. The solution was then stirred for 15 min and then sonicated for 20 min

  14. Innate stimulatory capacity of high molecular weight transition metals Au (gold) and Hg (mercury).

    Science.gov (United States)

    Rachmawati, Dessy; Alsalem, Inás W A; Bontkes, Hetty J; Verstege, Marleen I; Gibbs, Sue; von Blomberg, B M E; Scheper, Rik J; van Hoogstraten, Ingrid M W

    2015-03-01

    Nickel, cobalt and palladium ions can induce an innate immune response by triggering Toll-like receptor (TLR)-4 which is present on dendritic cells (DC). Here we studied mechanisms of action for DC immunotoxicity to gold and mercury. Next to gold (Na3Au (S2O3)2⋅2H2O) and mercury (HgCl2), nickel (NiCl2) was included as a positive control. MoDC activation was assessed by release of the pro-inflammatory mediator IL-8. Also PBMC were studied, and THP-1 cells were used as a substitution for DC for evaluation of cytokines and chemokines, as well as phenotypic, alterations in response to gold and mercury. Our results showed that both Na3Au (S2O3)2⋅2H2O and HgCl2 induce substantial release of IL-8, but not IL-6, CCL2 or IL-10, from MoDc, PBMC, or THP-1 cells. Also gold and, to a lesser extent mercury, caused modest dendritic cell maturation as detected by increased membrane expression of CD40 and CD80. Both metals thus show innate immune response capacities, although to a lower extent than reported earlier for NiCl2, CoCl2 and Na2 [PdCl4]. Importantly, the gold-induced response could be ascribed to TLR3 rather than TLR4 triggering, whereas the nature of the innate mercury response remains to be clarified. In conclusion both gold and mercury can induce innate immune responses, which for gold could be ascribed to TLR3 dependent signalling. These responses are likely to contribute to adaptive immune responses to these metals, as reflected by skin and mucosal allergies. Copyright © 2014 Elsevier Ltd. All rights reserved.

  15. High Molecular Weight Isoforms of Growth Hormone In Cells of the Immune System

    Science.gov (United States)

    Weigent, Douglas A.

    2013-01-01

    A substantial body of research exists to support the idea that cells of the immune system produce growth hormone (GH). However, the structure and mechanism of action of lymphocyte-derived GH continues to remain largely unknown. Here we present the results of Western analysis of whole cell extracts showing that different molecular weight isoforms of GH of approximately 100 kDa, 65 kDa, and 48 kDa can be detected in primary mouse cells of the immune system and in the mouse EL4 cell line. The identity of the 65 kDa and 48 kDa isoforms of GH were confirmed by mass spectrometry. The various isoforms were detected in both enriched T and B spleen cell populations. The large molecular weight isoform appears to reside primarily in the cytoplasm whereas the lower molecular weight 65 kDa and 48 kDa isoforms were detected primarily in the nucleus. These results also suggest that GH isoforms are induced by oxidative stress. In EL4 cells overexpressing GH, the expression of luciferase controlled by a promoter containing the antioxidant response element is increased almost three-fold above control. The data suggest that the induction of isoforms of the GH molecule in cells of the immune system may be an important mechanism of adaptation and/or protection of lymphoid cells under conditions of oxidative stress. PMID:21741628

  16. Split Nitrogen Application Improves Wheat Baking Quality by Influencing Protein Composition Rather Than Concentration.

    Science.gov (United States)

    Xue, Cheng; Auf'm Erley, Gunda Schulte; Rossmann, Anne; Schuster, Ramona; Koehler, Peter; Mühling, Karl-Hermann

    2016-01-01

    The use of late nitrogen (N) fertilization (N application at late growth stages of wheat, e.g., booting, heading or anthesis) to improve baking quality of wheat has been questioned. Although it increases protein concentration, the beneficial effect on baking quality (bread loaf volume) needs to be clearly understood. Two pot experiments were conducted aiming to evaluate whether late N is effective under controlled conditions and if these effects result from increased N rate or N splitting. Late N fertilizers were applied either as additional N or split from the basal N at late boot stage or heading in the form of nitrate-N or urea. Results showed that late N fertilization improved loaf volume of wheat flour by increasing grain protein concentration and altering its composition. Increasing N rate mainly enhanced grain protein quantitatively. However, N splitting changed grain protein composition by enhancing the percentages of gliadins and glutenins as well as certain high molecular weight glutenin subunits (HMW-GS), which led to an improved baking quality of wheat flour. The late N effects were greater when applied as nitrate-N than urea. The proportions of glutenin and x-type HMW-GS were more important than the overall protein concentration in determining baking quality. N splitting is more effective in improving wheat quality than the increase in the N rate by late N, which offers the potential to cut down N fertilization rates in wheat production systems.

  17. Split nitrogen application improves wheat baking quality by influencing protein composition rather than concentration

    Directory of Open Access Journals (Sweden)

    Cheng eXue

    2016-06-01

    Full Text Available The use of late nitrogen (N fertilization (N application at late growth stages of wheat, e.g. booting, heading or anthesis to improve baking quality of wheat has been questioned. Although it increases protein concentration, the beneficial effect on baking quality (bread loaf volume needs to be clearly understood. Two pot experiments were conducted aiming to evaluate whether late N is effective under controlled conditions and if these effects result from increased N rate or N splitting. Late N fertilizers were applied either as additional N or split from the basal N at late boot stage or heading in the form of nitrate-N or urea. Results showed that late N fertilization improved loaf volume of wheat flour by increasing grain protein concentration and altering its composition. Increasing N rate mainly enhanced grain protein quantitatively. However, N splitting changed grain protein composition by enhancing the percentages of gliadins and glutenins as well as certain high molecular weight glutenin subunits (HMW-GS, which led to an improved baking quality of wheat flour. The late N effects were greater when applied as nitrate-N than urea. The proportions of glutenin and x-type HMW-GS were more important than the overall protein concentration in determining baking quality. N splitting is more effective in improving wheat quality than the increase in the N rate by late N, which offers the potential to cut down N fertilization rates in wheat production systems.

  18. Subunit Stoichiometry of Human Muscle Chloride Channels

    OpenAIRE

    Fahlke, Christoph; Knittle, Timothy; Gurnett, Christina A.; Campbell, Kevin P.; George, Alfred L.

    1997-01-01

    Voltage-gated Cl? channels belonging to the ClC family appear to function as homomultimers, but the number of subunits needed to form a functional channel is controversial. To determine subunit stoichiometry, we constructed dimeric human skeletal muscle Cl? channels in which one subunit was tagged by a mutation (D136G) that causes profound changes in voltage-dependent gating. Sucrose-density gradient centrifugation experiments indicate that both monomeric and dimeric hClC-1 channels in their ...

  19. Role of the Rubisco Small Subunit

    Energy Technology Data Exchange (ETDEWEB)

    Spreitzer, Robert Joseph [Univ. of Nebraska, Lincoln, NE (United States)

    2016-11-05

    Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes the rate-limiting step of CO2 fixation in photosynthesis. However, it is a slow enzyme, and O2 competes with CO2 at the active site. Oxygenation initiates the photorespiratory pathway, which also results in the loss of CO2. If carboxylation could be increased or oxygenation decreased, an increase in net CO2 fixation would be realized. Because Rubisco provides the primary means by which carbon enters all life on earth, there is much interest in engineering Rubisco to increase the production of food and renewable energy. Rubisco is located in the chloroplasts of plants, and it is comprised of two subunits. Much is known about the chloroplast-gene-encoded large subunit (rbcL gene), which contains the active site, but much less is known about the role of the nuclear-gene-encoded small subunit in Rubisco function (rbcS gene). Both subunits are coded by multiple genes in plants, which makes genetic engineering difficult. In the eukaryotic, green alga Chlamydomonas reinhardtii, it has been possible to eliminate all the Rubisco genes. These Rubisco-less mutants can be maintained by providing acetate as an alternative carbon source. In this project, focus has been placed on determining whether the small subunit might be a better genetic-engineering target for improving Rubisco. Analysis of a variable-loop structure (βA-βB loop) of the small subunit by genetic selection, directed mutagenesis, and construction of chimeras has shown that the small subunit can influence CO2/O2 specificity. X-ray crystal structures of engineered chimeric-loop enzymes have indicated that additional residues and regions of the small subunit may also contribute to Rubisco function. Structural dynamics of the small-subunit carboxyl terminus was also investigated. Alanine-scanning mutagenesis of the most-conserved small-subunit residues has identified a

  20. The beta subunit of casein kinase II

    DEFF Research Database (Denmark)

    Boldyreff, B; Piontek, K; Schmidt-Spaniol, I

    1991-01-01

    cDNAs encoding the beta subunit of pig and mouse CKII were isolated. The porcine cDNA was expressed as a fusion protein in Escherichia coli and used for the production of anti-CKII-beta subunit specific antibodies....

  1. 28 CFR 51.6 - Political subunits.

    Science.gov (United States)

    2010-07-01

    ... 28 Judicial Administration 2 2010-07-01 2010-07-01 false Political subunits. 51.6 Section 51.6 Judicial Administration DEPARTMENT OF JUSTICE (CONTINUED) PROCEDURES FOR THE ADMINISTRATION OF SECTION 5 OF THE VOTING RIGHTS ACT OF 1965, AS AMENDED General Provisions § 51.6 Political subunits. All political...

  2. Acetylcholine Receptor: Complex of Homologous Subunits

    Science.gov (United States)

    Raftery, Michael A.; Hunkapiller, Michael W.; Strader, Catherine D.; Hood, Leroy E.

    1980-06-01

    The acetylcholine receptor from the electric ray Torpedo californica is composed of five subunits; two are identical and the other three are structurally related to them. Microsequence analysis of the four polypeptides demonstrates amino acid homology among the subunits. Further sequence analysis of both membrane-bound and Triton-solubilized, chromatographically purified receptor gave the stoichiometry of the four subunits (40,000:50,000:60,000:65,000 daltons) as 2:1:1:1, indicating that this protein is a pentameric complex with a molecular weight of 255,000 daltons. Genealogical analysis suggests that divergence from a common ancestral gene occurred early in the evolution of the receptor. This shared ancestry argues that each of the four subunits plays a functional role in the receptor's physiological action.

  3. The Subunit Principle in Scar Face Revision.

    Science.gov (United States)

    Elshahat, Ahmed; Lashin, Riham

    2017-06-01

    Facial scaring is considered one of the most difficult cosmetic problems for any plastic surgeon to solve. The condition is more difficult if the direction of the scar is not parallel to relaxed skin tension lines. Attempts to manage this difficult situation included revisions using geometric designs, Z plasties or W plasties to camouflage the straight line visible scaring. The use of long-lasting resorbable sutures was tried too. Recently, the use of botulinum toxin during revision improved the results. Fractional CO2 lasers, microfat grafts, and platelet-rich plasma were added to the armamentarium. The scar is least visible if placed in the junction between the facial subunits. The aim of this study is to investigate the use of the subunit principle to improve the results of scar revision. Four patients were included in this study. Tissue expansion of the intact part of the subunit allowed shifting the scar to the junction between the affected subunit and the adjacent one. Tissue expansion, delivery of the expanders, and advancement of the flaps were successful in all patients. The fact that this is a 2-stage procedure and sacrifices some of the intact skin from the affected facial subunit, makes this technique reserved to patients with ugly facial scars who are ambitious to improve their appearance.

  4. Baking quality parameters of wheat in relation to endosperm storage proteins

    Directory of Open Access Journals (Sweden)

    Daniela Horvat

    2012-01-01

    Full Text Available Wheat storage proteins of twelve winter wheat cultivars grown at the experimental field of the Agricultural Institute Osijek in 2009 were studied for their contribution to the baking quality. Composition of high molecular weight glutenin subunits (HMW-GS was analyzed by SDS-PAGE method, while the proportions of endosperm storage proteins were determined by RP-HPLC method. Regarding the proportion of storage proteins, results of the linear correlation (p<0.05 showed that protein (P and wet gluten (WG content were highly negatively correlated with albumins and globulins (AG and positively with α- gliadins (GLI. A strong negative correlation between AG and water absorption (WA capacity of flour was found, while α- GLI had positive influence on this property. Dough development time (DDT was positively significantly correlated with HMW-GS and negatively with AG. Degree of dough softening (DS was strongly positively affected by γ- GLI and gliadins to glutenins ratio (GLI/GLU and negatively by total GLU and HMW-GS. Dough energy (E and maximum resistance (RMAX were significantly positively affected by Glu-1 score and negatively by GLI/GLU ratio. Resistance to extensibility ratio (R/EXT was significantly negatively correlated with total GLI. Bread volume was significantly negatively influenced by AG.

  5. Development of a large-scale HPLC-based purification for the urease from Staphylococcus leei and determination of subunit structure.

    Science.gov (United States)

    Jin, Ming; Rosario, Wildys; Watler, Elsie; Calhoun, David H

    2004-03-01

    Coagulase-positive Staphylococcus species, related to but distinct from the genetic homology group containing Staphylococcus cohnii, Staphylococcus xylosus, and Staphylococcus saphrophyticus, were isolated from biopsy material obtained from a cluster of patients in Korea suffering from gastritis. The prototype isolate, Staphylococcus leei, has high urease activity that is similar with respect to a low K(m) value and acid resistance of the urease found in the stomach adapted pathogen, Helicobacter pylori. S. leei is remarkably resistant to lysis and only a small fraction of the cells are broken using sonication, a French press, Niro homogenizer, or a Gaulin mill. In the present report, we describe an efficient cell lysis procedure for S. leei using three passes through a Dynomill with 0.5mm glass beads that results in lysis of more than 95% of the cells. We also developed an efficient and large-scale purification procedure for the S. leei urease using a BioCAD HPLC Workstation using Q-Sepharose, Poros HP2, Sephacryl S-300, and hydroxyapatite chromatography. The urease of S. leei was purified 98-fold to a specific activity of 731U/mg. The urease protein is composed of three subunits, alpha (65kDa), beta (21kDa), and gamma (12kDa), and in situ enzyme assay and molecular sieve chromatography indicate that multiple high molecular weight forms are present, including an apparent pentamer of 1:1:1 alphabetagamma-heterotrimers of 480kDa. This purification procedure was used to purify 16mg of enzyme from 120-liters of cell culture. This improved lysis and purification procedure will make it possible to obtain sufficient quantities of urease for use as an antigen in ELISA assays to carry out studies to determine the incidence and demographic prevalence of gastritis due to S. leei.

  6. Immunochemical aspects of crotoxim and its subunits

    International Nuclear Information System (INIS)

    Nakazone, A.K.

    1979-01-01

    Crotamine and crotoxin with the subunits - phospholipase A and crotapotin - were obtained by purification from Crotalus durissus terrificus venom. Interaction studies of the subunits using crotalic antiserum, indicated that: crotoxin is formed of crotapotin and phospholipase A with the molar ratio of 1 to 1; using crotapotin 125 I the presence of a soluble complex was shown with the same antiserum. Immunological precipitation reactions demonstrated that crotapotin is antigenic: crotapotin and phospholipase A presented similar antigenic determinants; crotoxin antiserum reacted with each one of the submits; when the subunits are mixed to form synthetic crotoxin some antigenic determinants are masked in the process of interaction. Crotamine, interacted with crotapotin 1:1, without hidden antigenic determinants crotapotin antigenic site seems to be formed by, at least, one lysine. Enzimatical activity of phospholipase A apreared to be dependent on some reaction conditions when its arginine residues are blocked. Tyrosines of phospholipase A are more susceptible to labelling with 131 I than crotapotin. Gama irradiation of aqueous solutions of the subunits produced modifications in the ultraviolet spectra. A decrease of the enzymatic activity occured as a function of radiation dosis. Immunological activities of crotapotin and phospholipase A were not altered [pt

  7. Involvement of proteasomal subunits zeta and iota in RNA degradation.

    Science.gov (United States)

    Petit, F; Jarrousse, A S; Dahlmann, B; Sobek, A; Hendil, K B; Buri, J; Briand, Y; Schmid, H P

    1997-01-01

    We have identified two distinct subunits of 20 S proteasomes that are associated with RNase activity. Proteasome subunits zeta and iota, eluted from two-dimensional Western blots, hydrolysed tobacco mosaic virus RNA, whereas none of the other subunits degraded this substrate under the same conditions. Additionally, proteasomes were dissociated by 6 M urea, and subunit zeta, containing the highest RNase activity, was isolated by anion-exchange chromatography and gel filtration. Purified subunit zeta migrated as a single spot on two-dimensional PAGE with a molecular mass of approx. 28 kDa. Addition of anti-(subunit zeta) antibodies led to the co-precipitation of this proteasome subunit and nuclease activity. This is the first evidence that proteasomal alpha-type subunits are associated with an enzymic activity, and our results provide further evidence that proteasomes may be involved in cellular RNA metabolism. PMID:9337855

  8. Differential neutralizing activities of a single domain camelid antibody (VHH specific for ricin toxin's binding subunit (RTB.

    Directory of Open Access Journals (Sweden)

    Cristina Herrera

    Full Text Available Ricin, a member of the A-B family of ribosome-inactivating proteins, is classified as a Select Toxin by the Centers for Disease Control and Prevention because of its potential use as a biothreat agent. In an effort to engineer therapeutics for ricin, we recently produced a collection of alpaca-derived, heavy-chain only antibody VH domains (VHH or "nanobody" specific for ricin's enzymatic (RTA and binding (RTB subunits. We reported that one particular RTB-specific VHH, RTB-B7, when covalently linked via a peptide spacer to different RTA-specific VHHs, resulted in heterodimers like VHH D10/B7 that were capable of passively protecting mice against a lethal dose challenge with ricin. However, RTB-B7 itself, when mixed with ricin at a 1 ∶ 10 toxin:antibody ratio did not afford any protection in vivo, even though it had demonstrable toxin-neutralizing activity in vitro. To better define the specific attributes of antibodies associated with ricin neutralization in vitro and in vivo, we undertook a more thorough characterization of RTB-B7. We report that RTB-B7, even at 100-fold molar excess (toxin:antibody was unable to alter the toxicity of ricin in a mouse model. On the other hand, in two well-established cytotoxicity assays, RTB-B7 neutralized ricin with a 50% inhibitory concentration (IC50 that was equivalent to that of 24B11, a well-characterized and potent RTB-specific murine monoclonal antibody. In fact, RTB-B7 and 24B11 were virtually identical when compared across a series of in vitro assays, including adherence to and neutralization of ricin after the toxin was pre-bound to cell surface receptors. RTB-B7 differed from both 24B11 and VHH D10/B7 in that it was relatively less effective at blocking ricin attachment to receptors on host cells and was not able to form high molecular weight toxin:antibody complexes in solution. Whether either of these activities is important in ricin toxin neutralizing activity in vivo remains to be determined.

  9. Genetic evolution and utilization of wheat germplasm resources in Huanghuai winter wheat region of China

    International Nuclear Information System (INIS)

    Xiyong, C.; Haixia, X.U.; Feng, C.

    2011-01-01

    To determine the genetic variation of wheat germplasm resources and improve their use in wheat breeding, 215 wheat cultivars and advanced lines from the Huanghuai Wheat Region of China were used to identify 14 agronomic traits and 7 quality traits, as well as the evolution and utilization of high molecular weight glutenin subunits (HMW-GS) and low molecular weight-glutenin subunits (LMW-GS). From land race cultivars to current cultivars there had been significant increases in grain numbers spike/sip -1/, grain weight spike/sup -1/, 1000-kernel weight, grain weight plant/sup -1/, spikelet number spike/sup -1/, sterile spikelet numbers spike/sup -1/, flag leaf width, and flag leaf area. There had been significant decreases in spike number plant/sup -1/, plant height, the first inter node length, flag leaf length, kernel protein content and wet gluten content. Based on Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) results, a novel HMW-GS combination 20/8 was identified in 1B chromosome of Chinese landrace cultivar Heputou. Subunits 22, 20/8, 2.2+12, and GluB3a were only found in cultivars before the 1960s, and subunits 6+8, 13+16, 3+12, and 4+12 were only found in the cultivars after the 1980s. The average diversity index of 21 traits and allele variance of HMW-GS showed a decreasing-increasing-decreasing tendency. HMW-GS and LMW-GS combination-type cultivars showed an increasing-decreasing tendency. Before the 1980s, most parental strains were from foreign cultivars and landrace cultivars, while after the 1980s, most parental strains were from released cultivars and germplasm created by distant hybridization. This study provided useful information for improvement of wheat breeding in Huanghuai winter wheat region. (author)

  10. Mechanistic and morphological origins of ultra-high molecular weight polyethylene wear debris in total joint replacement prostheses.

    Science.gov (United States)

    Wang, A; Stark, C; Dumbleton, J H

    1996-01-01

    The mechanistic and morphological origins of microscopic wear debris generated from UHMWPE articular surfaces in total joint replacement prostheses are investigated in this study. It was found experimentally that the molecular chain structure at the articulating surface of UHMWPE undergoes a re-organization process due to strain accumulation caused by surface traction. This molecular re-organization process creates a fibre-like surface texture that exhibits an anisotropic behaviour similar to a unidirectionally reinforced polymer composite. This composite responds to stresses favourably if loaded along the fibre axis but unfavourably if loaded off axis. Due to the very complex multi-axial motion/loading nature at the articular surfaces in total joint replacements, the stress tensors applied to each localized asperity contact area continuously change their directions and magnitudes. These changes in the localized stress field create an off-axis loading situation at each localized contact zone with respect to the orientation of the molecular chains. Depending on the off-axis angle, failure of the molecular structure occurs in three different ways: tensile rupture at very small off-axis angles, shear rupture at intermediate off-axis angles and transverse splitting at large off-axis angles. These failure mechanisms all produce similar fibre-like wear debris. However, the failure stresses differ significantly among the three modes. According to this molecular wear theory, the preferred polymer microstructure for optimal wear resistance would be a three-dimensionally strong network connected by covalent bonds between molecular chains. For UHMWPE, a three-dimensional molecular network can be created by radiation induced cross-linking. Experiments conducted on both gamma irradiated and unirradiated UHMWPE specimens using a linear wear machine and multi-axial joint simulators confirmed the validity of the molecular wear theory.

  11. A new high molecular weight immunoglobulin class from the carcharhine shark: implications for the properties of the primordial immunoglobulin.

    Science.gov (United States)

    Berstein, R M; Schluter, S F; Shen, S; Marchalonis, J J

    1996-04-16

    All immunoglobulins and T-cell receptors throughout phylogeny share regions of highly conserved amino acid sequence. To identify possible primitive immunoglobulins and immunoglobulin-like molecules, we utilized 3' RACE (rapid amplification of cDNA ends) and a highly conserved constant region consensus amino acid sequence to isolate a new immunoglobulin class from the sandbar shark Carcharhinus plumbeus. The immunoglobulin, termed IgW, in its secreted form consists of 782 amino acids and is expressed in both the thymus and the spleen. The molecule overall most closely resembles mu chains of the skate and human and a new putative antigen binding molecule isolated from the nurse shark (NAR). The full-length IgW chain has a variable region resembling human and shark heavy-chain (VH) sequences and a novel joining segment containing the WGXGT motif characteristic of H chains. However, unlike any other H-chain-type molecule, it contains six constant (C) domains. The first C domain contains the cysteine residue characteristic of C mu1 that would allow dimerization with a light (L) chain. The fourth and sixth domains also contain comparable cysteines that would enable dimerization with other H chains or homodimerization. Comparison of the sequences of IgW V and C domains shows homology greater than that found in comparisons among VH and C mu or VL, or CL thereby suggesting that IgW may retain features of the primordial immunoglobulin in evolution.

  12. High-molecular-weight polymers containing biodegradable disulfide bonds: synthesis and in vitro verification of intracellular degradation

    Czech Academy of Sciences Publication Activity Database

    Etrych, Tomáš; Kovář, Lubomír; Šubr, Vladimír; Braunová, Alena; Pechar, Michal; Chytil, Petr; Říhová, Blanka; Ulbrich, Karel

    2010-01-01

    Roč. 25, č. 1 (2010), s. 5-26 ISSN 0883-9115 R&D Projects: GA AV ČR IAA400500806; GA AV ČR KAN200200651 Institutional research plan: CEZ:AV0Z40500505; CEZ:AV0Z50200510 Keywords : water-soluble polymers * reductive degradation * HPMA copolymers Subject RIV: CD - Macromolecular Chemistry Impact factor: 2.610, year: 2010

  13. Healing of interfaces of high and ultra-high-molecular- weight polystyrene below the bulk glass transition temperature

    DEFF Research Database (Denmark)

    Boiko, Yuri M.; Lyngaae-Jørgensen, Jørgen

    2004-01-01

    into contact to themselves below the glass transition temperature T-g of the bulk Tg-bulk, in a lap-shear joint geometry, at a constant healing temperature T-h for a healing time t(h) of 10 min to 24 h. The lap-shear strength sigma of the symmetric HMWPS-HMWPS and UHMWPS-URMWPS interfaces has been measured...

  14. Immunological identification of a high molecular weight protein as a candidate for the product of the Duchenne muscular dystrophy gene.

    OpenAIRE

    Kao, L; Krstenansky, J; Mendell, J; Rammohan, K W; Gruenstein, E

    1988-01-01

    An oligopeptide was synthesized based on translation of the nucleotide sequence of the putative exon region of clone pERT87-25 from the gene for Duchenne muscular dystrophy. Immunization of rabbits with this oligopeptide induced the formation of antibodies directed against a protein present in human, rat, and rabbit skeletal muscle. This protein, which is missing in the skeletal muscle of two patients with Duchenne muscular dystrophy, has a molecular mass of approximately equal to 320-420 kDa...

  15. Absence of differences among low, middle, and high molecular weight hyaluronan in activating murine immune cells in vitro

    Czech Academy of Sciences Publication Activity Database

    Šafránková, Barbora; Hermannová, M.; Nešporová, K.; Velebný, V.; Kubala, Lukáš

    2018-01-01

    Roč. 107, FEB2018 (2018), s. 1-8 ISSN 0141-8130 Institutional support: RVO:68081707 Keywords : mouse macrophages * chondroitin sulfate * prostaglandin e-2 * gene-expression Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Cell biology Impact factor: 3.671, year: 2016

  16. Comparison of Low-, Moderate-, and High-Molecular-Weight Hyaluronic Acid Injections in Delaying Time to Knee Surgery.

    Science.gov (United States)

    Shewale, Anand R; Barnes, Charles L; Fischbach, Lori A; Ounpraseuth, Songthip T; Painter, Jacob T; Martin, Bradley C

    2017-10-01

    We compared the effectiveness of low-molecular-weight (MW) hyaluronic acid (HA) injections (LMWHA), moderate-MW HA injections (MMWHA), and high-MW HA injections (HMWHA) for prevention or delay of knee surgery in patients with knee osteoarthritis. An observational cohort study using LifeLink Plus claims (2006-2015) was used. The primary outcome measure of the study included all surgical interventions of the knee. The secondary outcome measures were the following: (1) unicompartmental knee arthroplasty or total knee arthroplasty and (2) total knee arthroplasty only. A high-dimensional propensity score (hdPS) using 1:1 matching was used to adjust for confounding. The likelihood of each outcome was assessed using Cox proportional hazard models. A cohort of 30,417 incident HA users with knee osteoarthritis met our inclusion-exclusion criteria. There was no difference in the likelihood of composite surgical events between LMWHA users (hazard ratio, 0.939; 95% confidence interval, 0.870-1.013) and MMWHA users (hazard ratio, 1.032; 95% confidence interval, 0.952-1.119) when compared with HMWHA users in a matched hdPS analysis. However, a significantly lower likelihood for all outcome measures was demonstrated in LMWHA and MMWHA users compared with HMWHA users when hdPS was not used. There was no significant difference in the likelihood of surgical interventions between LMWHA, MMWHA, and HMWHA users after accounting for empirically derived confounders. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Occurrence of tetrodotoxin-binding high molecular weight substances in the body fluid of shore crab (Hemigrapsus sanguineus).

    Science.gov (United States)

    Shiomi, K; Yamaguchi, S; Kikuchi, T; Yamamori, K; Matsui, T

    1992-12-01

    The shore crab (Hemigrapsus sanguineus) is highly resistant to tetrodotoxin (TTX) although it contains no detectable amount of TTX (less than 5 MU/g, where 1 MU is defined as the amount of TTX killing a 20 g mouse in 30 min). Its body fluid was examined for neutralizing effects against the lethal activity of TTX. When the mixture of the body fluid and TTX was injected i.p. into mice, the lethal activity of TTX was significantly reduced; 1 ml of the body fluid was evaluated to neutralize 3.6-4.0 MU of TTX. Higher neutralizing activity (7.2-12.5 MU/ml of the body fluid) was exhibited by i.v. administration of the body fluid into mice before or after i.p. challenge of TTX. The lethal effect of paralytic shellfish poisons was not counteracted by the body fluid. Analysis by gel filtration on Sepharose 6B revealed that the body fluid contained TTX-binding high mol. wt substances (> 2,000,000) responsible for the neutralizing activity of the body fluid against TTX, which accounts for the high resistibility of the crab to TTX. When the crude toxin extracted from the liver of puffer (Takifugu niphobles) was mixed with the body fluid and chromatographed on Sepharose 6B, almost pure TTX was obtained from the fractions containing the TTX-binding high mol. wt substances, suggesting that the TTX-binding high mol. wt substances could be useful in purification of TTX from biological samples.

  18. Simple colorimetric methods for determination of sub-milligram amounts of ultra-high molecular weight polyethylene wear particles

    Czech Academy of Sciences Publication Activity Database

    Veselý, F.; Zolotarevova, E.; Špundová, M.; Kaftan, Filip; Šlouf, Miroslav; Entlicher, G.

    2012-01-01

    Roč. 8, č. 5 (2012), s. 1935-1938 ISSN 1742-7061 R&D Projects: GA MŠk 2B06096; GA MZd NT12229 Grant - others:GA ČR(CZ) GAP503/11/0163 Institutional research plan: CEZ:AV0Z40550506; CEZ:AV0Z40500505 Keywords : joint replacement * polyethylene wear particles * wear particles determination * colorimetric methods Subject RIV: CC - Organic Chemistry Impact factor: 5.093, year: 2012

  19. Characterization of a novel fructosyltransferase from Lactobacillus reuteri that synthesizes high-molecular-weight inulin and inulin oligosaccharides

    NARCIS (Netherlands)

    Hijum, S.A.F.T. van; Geel-Schutten, G.H. van; Rahaoui, H.; Maarel, M.J.E.C. van der; Dijkhuizen, L.

    Fructosyltransferase (FTF) enzymes produce fructose polymers (fructans) from sucrose. Here, we report the isolation and characterization of an FTF-encoding gene from Lactobacillus reuteri strain 121. A C-terminally truncated version of the ftf gene was successfully expressed in Escherichia coli.

  20. Immunological identification of a high molecular weight protein as a condidate for the product of the Duchenne muscular dystrophy gene

    Energy Technology Data Exchange (ETDEWEB)

    Kao, L.; Krstenansky, J.; Mendell, J.; Rammohan, K.W.; Gruenstein, E. (Univ. of Cincinnati College of Medicine, OH (USA))

    1988-06-01

    An oligopeptide was synthesized based on translation of the nucleotide sequence of the putative exon region of clone pERT87-25 from the gene for Duchenne muscular dystrophy. Immunization of rabbits with this oligopeptide induced the formation of antibodies directed against a protein present in human, rat, and rabbit skeletal muscle. This protein, which is missing in the skeletal muscle of two patients with Duchenne muscular dystrophy, has a molecular mass of {approx}320-420 kDa and is clearly different from the putative Duchenne muscular dystrophy-related protein nebulin. The data suggest that this 320-420-kDa protein is produced by the Duchenne muscular dystrophy gene.

  1. Effects of D-002, a mixture of high molecular weight beeswax alcohols, on patients with nonalcoholic fatty liver disease.

    Science.gov (United States)

    Illnait, José; Rodríguez, Iván; Mendoza, Sarahí; Fernández, Yolanda; Mas, Rosa; Miranda, Mirtha; Piñera, Jesús; Fernández, Julio César; Mesa, Meilis; Fernández, Lilia; Carbajal, Daisy; Gámez, Rafael

    2013-07-01

    Nonalcoholic fatty liver disease (NAFLD) is intimately related to insulin resistance and ranges from a benign course to liver fibrosis and cirrhosis. NAFLD management mainly involves dietary modification and weight loss. Although no fully successful pharmacological intervention is available, alternative therapies to treat NAFLD have shown promising results. Experimental studies have shown that D-002, a mixture of beeswax alcohols with antioxidant effects, is hepatoprotective. The aim of this study was to investigate the efficacy and safety of D-002 in patients with NALFD. Fifty patients with NAFLD were randomized to receive a placebo or D-002 (100 mg/day) for 24 weeks. The primary endpoint was a significant ultrasonography-detected reduction of liver fat infiltration versus a placebo. Secondary endpoints were decreases in the homeostatic model assessment (HOMA) index, insulin levels, serum liver enzymes, increases in plasma total antioxidant status (TAS) and improved clinical symptoms versus the placebo recipients. At randomization, all indicators were comparable in both groups. At study completion, seven (28.0%) D-002-patients, but none of the placebo recipients, exhibited a normal liver echo pattern on ultrasonography (p < 0.01). Also, D-002 significantly reduced (p < 0.01 vs. baseline and placebo) the HOMA index and insulin levels and increased the TAS, but did not affect other parameters. The proportion of D-002-patients (12/25, 48.0%) showing symptom improvement was higher (p < 0.001) than that of the placebo group (1/25, 4.0%). The treatment was safe and well tolerated. Three patients in each group withdrew from the study. D-002 (100 mg/day) improved ultrasonographic findings, indicators of insulin resistance, plasma TAS and clinical evolution on NAFLD patients. Further studies, however, are needed to confirm these results.

  2. A Modified ELISA Accurately Measures Secretion of High Molecular Weight Hyaluronan (HA) by Graves' Disease Orbital Cells

    Science.gov (United States)

    Krieger, Christine C.

    2014-01-01

    Excess production of hyaluronan (hyaluronic acid [HA]) in the retro-orbital space is a major component of Graves' ophthalmopathy, and regulation of HA production by orbital cells is a major research area. In most previous studies, HA was measured by ELISAs that used HA-binding proteins for detection and rooster comb HA as standards. We show that the binding efficiency of HA-binding protein in the ELISA is a function of HA polymer size. Using gel electrophoresis, we show that HA secreted from orbital cells is primarily comprised of polymers more than 500 000. We modified a commercially available ELISA by using 1 million molecular weight HA as standard to accurately measure HA of this size. We demonstrated that IL-1β-stimulated HA secretion is at least 2-fold greater than previously reported, and activation of the TSH receptor by an activating antibody M22 from a patient with Graves' disease led to more than 3-fold increase in HA production in both fibroblasts/preadipocytes and adipocytes. These effects were not consistently detected with the commercial ELISA using rooster comb HA as standard and suggest that fibroblasts/preadipocytes may play a more prominent role in HA remodeling in Graves' ophthalmopathy than previously appreciated. PMID:24302624

  3. Anticoagulant and calcium-binding properties of high molecular weight derivatives of human fibrinogen (plasmin fragments Y)

    NARCIS (Netherlands)

    Nieuwenhuizen, W.; Voskuilen, M.; Hermans, J.

    1982-01-01

    The present study was undertaken as a step to delineate further the localization of the calcium-binding sites in fibrinogen and to assess the anticlotting properties of fibrinogen degradation products. To this purpose, fragments Y were prepared by plasmin digestion of human fibrinogen in the

  4. Anticoagulant and calcium-binding properties of high molecular weight derivatives of human fibrinogen, produced by plasmin (fragments X)

    NARCIS (Netherlands)

    Nieuwenhuizen, W.; Gravesen, M.

    1981-01-01

    Early plasmin degradation products (X fragments) of human fibrinogen were prepared in the presence of calcium-ions or EGTA, and purified on Sepharose 6B-CL. X fragments were characterized with respect to amino-terminal amino acids, polypeptide-chain composition, anticlotting properties and

  5. The extraction of zirconium (IV) from sulfuric acid solutions with high-molecular weight quaternary ammonium compound

    International Nuclear Information System (INIS)

    Sato, Taichi; Watanabe, Hiroshi

    1982-01-01

    The extraction of zirconium sulfate in aqueous sulfuric acid solutions with trioctylmethylammonium chloride (Aliquat-336; R 3 R'NCl) in organic solvents has been investigated under different conditions. In addition, the organic phases extracted sulfuric acid and zirconium sulfate were examined by IR and NMR spectroscopies. It has been found that Aliquat-336 extracts zirconium (IV) from sulfuric acid solutions according to the following ion-exchange reactions. i) The extraction of sulfuric acid is at first carried out through the equilibria, SO 4 2 - (aq) + 2R 3 R'NCl(org) reversible (R 3 R'N) 2 SO 4 (org) + 2Cl - (aq), (R 3 R'N) 2 SO 4 (org) + H + (aq) + HSO 4- (aq) reversible 2R 3 R'NHSO 4 (org). ii) The extraction of zirconium is expressed as the equilibrium reaction, Zr(SO 4 ) 3 2 - (aq) + 2xR 3 R'NHSO 4 (org) + (1-x)(R 3 R'N) 2 SO 4 (org) reversible (R 3 R'N) 2 [Zr(SO 4 ) 3 ](org) + xH 2 SO 4 (aq) + SO 4 2 - (aq), x = [R 3 R'NHSO 4 ]/(2[(R 3 R'N) 2 SO 4 ] + [R 3 R'NHSO 4 ]). Moreover, the hydrolyzed species (R 3 R'N)[ZrO(OH)(SO 4 )] is formed when zirconium is further extracted in an organic phase. (author)

  6. Origin of organic sulphur compounds and sulphur-containing high molecular weight substances in sediments and immature crude oils

    NARCIS (Netherlands)

    Sinninghe Damsté, J.S.; Rijpstra, W.I.C.; Leeuw, J.W. de; Schenck, P.A.

    1988-01-01

    The distribution patterns of Organic Sulphur Compounds (OSC), occurring in certain sediments and immature crude oils, were compared with those of the corresponding hydrocarbons. Because of the complexity of the OSC mixtures, they were desulphurized to hydrocarbons (n-alkanes, isoprenoid alkanes,

  7. Mechanical, Rheological, and Bioactivity Properties of Ultra High-Molecular-Weight Polyethylene Bioactive Composites Containing Polyethylene Glycol and Hydroxyapatite

    Directory of Open Access Journals (Sweden)

    Mazatusziha Ahmad

    2012-01-01

    Full Text Available Ultrahigh-molecular-weight polyethylene/high-density polyethylene (UHMWPE/HDPE blends prepared using polyethylene glycol PEG as the processing aid and hydroxyapatite (HA as the reinforcing filler were found to be highly processable using conventional melt blending technique. It was demonstrated that PEG reduced the melt viscosity of UHMWPE/HDPE blend significantly, thus improving the extrudability. The mechanical and bioactive properties were improved with incorporation of HA. Inclusion of HA from 10 to 50 phr resulted in a progressive increase in flexural strength and modulus of the composites. The strength increment is due to the improvement on surface contact between the irregular shape of HA and polymer matrix by formation of mechanical interlock. The HA particles were homogenously distributed even at higher percentage showed improvement in wetting ability between the polymer matrix and HA. The inclusion of HA enhanced the bioactivity properties of the composite by the formation of calcium phosphate (Ca-P precipitates on the composite surface as proven from SEM and XRD analysis.

  8. The interplay of plasma treatment and gold coating and ultra-high molecular weight polyethylene: On the cytocompatibility

    Czech Academy of Sciences Publication Activity Database

    Novotná, Z.; Rimpelová, S.; Juřík, P.; Veselý, M.; Kolská, Z.; Hubáček, Tomáš; Ruml, T.; Švorčík, V.

    2017-01-01

    Roč. 71, FEB (2017), s. 125-131 ISSN 0928-4931 R&D Projects: GA MŠk LM2015075 Institutional support: RVO:60077344 Keywords : UHMWPE * plasma treatment * gold sputtering * atomic force microscopy Subject RIV: JJ - Other Materials OBOR OECD: Materials engineering Impact factor: 4.164, year: 2016

  9. The high molecular weight dipeptidyl peptidase IV Pol d 3 is a major allergen of Polistes dominula venom

    DEFF Research Database (Denmark)

    Schiener, Maximilian; Hilger, Christiane; Eberlein, Bernadette

    2018-01-01

    Hymenoptera venom allergy can cause severe anaphylaxis in untreated patients. Polistes dominula is an important elicitor of venom allergy in Southern Europe as well as in the United States. Due to its increased spreading to more moderate climate zones, Polistes venom allergy is likely to gain imp...

  10. The high molecular weight dipeptidyl peptidase IV Pol d 3 is a major allergen of Polistes dominula venom.

    Science.gov (United States)

    Schiener, Maximilian; Hilger, Christiane; Eberlein, Bernadette; Pascal, Mariona; Kuehn, Annette; Revets, Dominique; Planchon, Sébastien; Pietsch, Gunilla; Serrano, Pilar; Moreno-Aguilar, Carmen; de la Roca, Federico; Biedermann, Tilo; Darsow, Ulf; Schmidt-Weber, Carsten B; Ollert, Markus; Blank, Simon

    2018-01-22

    Hymenoptera venom allergy can cause severe anaphylaxis in untreated patients. Polistes dominula is an important elicitor of venom allergy in Southern Europe as well as in the United States. Due to its increased spreading to more moderate climate zones, Polistes venom allergy is likely to gain importance also in these areas. So far, only few allergens of Polistes dominula venom were identified as basis for component-resolved diagnostics. Therefore, this study aimed to broaden the available panel of important Polistes venom allergens. The 100 kDa allergen Pol d 3 was identified by mass spectrometry and found to be a dipeptidyl peptidase IV. Recombinantly produced Pol d 3 exhibited sIgE-reactivity with approximately 66% of Polistes venom-sensitized patients. Moreover, its clinical relevance was supported by the potent activation of basophils from allergic patients. Cross-reactivity with the dipeptidyl peptidases IV from honeybee and yellow jacket venom suggests the presence of exclusive as well as conserved IgE epitopes. The obtained data suggest a pivotal role of Pol d 3 as sensitizing component of Polistes venom, thus supporting its status as a major allergen of clinical relevance. Therefore, Pol d 3 might become a key element for proper diagnosis of Polistes venom allergy.

  11. Stripping of 1.04 MeV per nucleon krypton ions in high molecular weight vapours

    International Nuclear Information System (INIS)

    Eastham, D.A.; Joy, T.; Clark, R.B.; King, R.

    1976-01-01

    Equilibrium charge state distributions have been measured for 1.04 MeV per nucleon krypton ions in heavy vapours with molecular weights from 462 to 6500. Non-equilibrium data are presented for the heaviest vapour. A maximum increase of 0.8 in the mean charge is found relative to a conventional diatomic gas but the pressures required are two orders of magnitude less. (Auth.)

  12. Specific labeling and assignment strategies of valine methyl groups for NMR studies of high molecular weight proteins

    Energy Technology Data Exchange (ETDEWEB)

    Mas, Guillaume; Crublet, Elodie [Univ. Grenoble Alpes, Institut de Biologie Structurale (IBS) (France); Hamelin, Olivier [CNRS (France); Gans, Pierre; Boisbouvier, Jérôme, E-mail: jerome.boisbouvier@ibs.fr [Univ. Grenoble Alpes, Institut de Biologie Structurale (IBS) (France)

    2013-09-28

    The specific protonation of valine and leucine methyl groups in proteins is typically achieved by overexpressing proteins in M9/D{sub 2}O medium supplemented with either labeled α-ketoisovalerate for the labeling of the four prochiral methyl groups or with 2-acetolactate for the stereospecific labeling of the valine and leucine side chains. However, when these labeling schemes are applied to large protein assemblies, significant overlap between the correlations of the valine and leucine methyl groups occurs, hampering the analysis of 2D methyl-TROSY spectra. Analysis of the leucine and valine biosynthesis pathways revealed that the incorporation of labeled precursors in the leucine pathway can be inhibited by the addition of exogenous l-leucine-d{sub 10}. We exploited this property to label stereospecifically the pro-R and pro-S methyl groups of valine with minimal scrambling to the leucine residues. This new labeling protocol was applied to the 468 kDa homododecameric peptidase TET2 to decrease the complexity of its NMR spectra. All of the pro-S valine methyl resonances of TET2 were assigned by combining mutagenesis with this innovative labeling approach. The assignments were transferred to the pro-R groups using an optimally labeled sample and a set of triple resonance experiments. This improved labeling scheme enables us to overcome the main limitation of overcrowding in the NMR spectra of prochiral methyl groups, which is a prerequisite for the site-specific measurement of the structural and dynamic parameters or for the study of interactions in very large protein assemblies.

  13. Studies on electronic spectrum and electron spin resonance of vanadium (IV) complexes with organophosphorus compounds and high molecular weight amines

    International Nuclear Information System (INIS)

    Sato, Taichi; Nakamura, Takato

    1981-01-01

    In the extraction of vanadium (IV) from aqueous solutions containing hydrochloric acid and/or a mixture of hydrochloric acid and lithium chloride by bis(2-ethylhexyl) hydrogenphosphate (DEHPA; HX), trioctylmethylammonium chloride (Aliquat-336), trioctylamine (TOA), trioctylphosphine oxide (TOPO) and tributyl phosphate (TBP), the complexes formed in the organic phases have been examined by spectrophotometry and electron spin resonance spectroscopy. It is found that in the extraction by DEHPA, the vanadium in the organic phase exists as the monomeric species, VO(X 2 H) 2 , or the polymeric one, (VOX 2 )sub(n), and that in the extractions by Aliquat-336, TOA, TOPO, and TBP, tetravalent vanadium complexes are stable in the organic phases extracted from a mixed solution of hydrochloric acid and lithium chloride, while complexes containing pentavalent vanadium and VOV 4+ ions are formed in the organic phases extracted from hydrochloric acid solutions. (author)

  14. Extraction chromatography of indium (III) on silica gel impregnated with high molecular weight carboxylic acid and its analytical applications

    International Nuclear Information System (INIS)

    Majumdar, P.S.; Ray, U.S.

    1991-01-01

    Indium(III) was separated by extraction chromatography with Versatic 10 as a stationary phase on a column of silica gel from acetic acid and sodium acetate solution (pH 4.5-6.0). The optimum condition for extraction was studied based on the critical study of the relevant factors as effects of pH, flow rate on extraction and elution. Role of stripping agents on the elution was studied. The separation of indium from a number of elements was carried out. Indium(III) was separated from Alsup(III), Gasup(III), Tlsup(III), Zrsup(IV) and trivalent lanthanides which interfere under the recommended extraction condition by exploiting the differences in their stripping behaviour. (author). 7 refs., 1 tab., 1 fig

  15. High molecular weight PEGylation of human pancreatic polypeptide at position 22 improves stability and reduces food intake in mice

    DEFF Research Database (Denmark)

    Thieme, V; Jolly, N; Madsen, A N

    2016-01-01

    BACKGROUND AND PURPOSE: Human pancreatic polypeptide (hPP) is known to suppress appetite and food intake, thereby representing a potential therapeutic approach against obesity and associated metabolic disorders. The aim of this study was to improve hPP stability by covalent PEGylation with diverse...... fasting-induced food intake and bioavailability. KEY RESULTS: In human epithelia and colonic mucosal preparations, activity of the modified hPP peptides depended on the core sequence and latency of the peptides was related to PEG size. Peptides modified with a 22 kDa PEG (PEG22) remained intact in blood...... plasma and on incubation with liver homogenates for more than 96 h. Finally, hPP2-36 , [K(22) (PEG22)]hPP2-36 and [K(22) (PEG22),Q(34) ]hPP significantly reduced cumulative food intake in mice over 16 h after s.c. administration. CONCLUSIONS AND IMPLICATIONS: Modification with PEG22 at position 22...

  16. A human scFv antibody that targets and neutralizes high molecular weight pathogenic amyloid-β oligomers.

    Science.gov (United States)

    Sebollela, Adriano; Cline, Erika N; Popova, Izolda; Luo, Kevin; Sun, Xiaoxia; Ahn, Jay; Barcelos, Milena A; Bezerra, Vanessa N; Lyra E Silva, Natalia M; Patel, Jason; Pinheiro, Nathalia R; Qin, Lei A; Kamel, Josette M; Weng, Anthea; DiNunno, Nadia; Bebenek, Adrian M; Velasco, Pauline T; Viola, Kirsten L; Lacor, Pascale N; Ferreira, Sergio T; Klein, William L

    2017-07-03

    Brain accumulation of soluble oligomers of the amyloid-β peptide (AβOs) is increasingly considered a key early event in the pathogenesis of Alzheimer's disease (AD). A variety of AβO species have been identified, both in vitro and in vivo, ranging from dimers to 24mers and higher order oligomers. However, there is no consensus in the literature regarding which AβO species are most germane to AD pathogenesis. Antibodies capable of specifically recognizing defined subpopulations of AβOs would be a valuable asset in the identification, isolation, and characterization of AD-relevant AβO species. Here, we report the characterization of a human single chain antibody fragment (scFv) denoted NUsc1, one of a number of scFvs we have identified that stringently distinguish AβOs from both monomeric and fibrillar Aβ. NUsc1 readily detected AβOs previously bound to dendrites in cultured hippocampal neurons. In addition, NUsc1 blocked AβO binding and reduced AβO-induced neuronal oxidative stress and tau hyperphosphorylation in cultured neurons. NUsc1 further distinguished brain extracts from AD-transgenic mice from wild type (WT) mice, and detected endogenous AβOs in fixed AD brain tissue and AD brain extracts. Biochemical analyses indicated that NUsc1 targets a subpopulation of AβOs with apparent molecular mass greater than 50 kDa. Results indicate that NUsc1 targets a particular AβO species relevant to AD pathogenesis, and suggest that NUsc1 may constitute an effective tool for AD diagnostics and therapeutics. © 2017 International Society for Neurochemistry.

  17. [Treatment of acute full-thickness chondral defects with high molecular weight hyaluronic acid; an experimental model].

    Science.gov (United States)

    Figueroa, D; Espinosa, M; Calvo, R; Scheu, M; Valderrama, J J; Gallegos, M; Conget, P

    2014-01-01

    To evaluate the effect of 2 different protocols of intra-articular hyaluronic acid (HA, hylan G-F20) to articular cartilage regeneration in acute full-thickness chondral defects. Full-thickness chondral defects of 3 x 6 mm were performed into the lateral femoral condyles of New Zealand rabbits, treated with a single or three doses of HA. The animals were sacrified at 12 weeks and the regenerated tissue was evaluated by direct observation and histology with the ICRS scale. Macroscopically, in both groups treated with HA the defects were filled with irregular tissue with areas similar to hyaline cartilage and others in which depressed areas with exposed subchondral bone were observed. Histological analysis showed in both groups treated with HA a hyaline-like cartilage compared to control group. However, the score of the International Cartilage Repair Society (ICRS) scale did not show differences between the groups treated with HA. The use of single dose or 3 doses of AH in acute chondral lesions has a limited and similar benefit in articular cartilage regeneration. Copyright © 2014 SECOT. Published by Elsevier Espana. All rights reserved.

  18. Immunological identification of a high molecular weight protein as a condidate for the product of the Duchenne muscular dystrophy gene

    International Nuclear Information System (INIS)

    Kao, L.; Krstenansky, J.; Mendell, J.; Rammohan, K.W.; Gruenstein, E.

    1988-01-01

    An oligopeptide was synthesized based on translation of the nucleotide sequence of the putative exon region of clone pERT87-25 from the gene for Duchenne muscular dystrophy. Immunization of rabbits with this oligopeptide induced the formation of antibodies directed against a protein present in human, rat, and rabbit skeletal muscle. This protein, which is missing in the skeletal muscle of two patients with Duchenne muscular dystrophy, has a molecular mass of ∼320-420 kDa and is clearly different from the putative Duchenne muscular dystrophy-related protein nebulin. The data suggest that this 320-420-kDa protein is produced by the Duchenne muscular dystrophy gene

  19. A Highly Expressed High-Molecular-Weight S-Layer Complex of Pelosinus sp. Strain UFO1 Binds Uranium

    Energy Technology Data Exchange (ETDEWEB)

    Thorgersen, Michael P. [Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology; Lancaster, W. Andrew [Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology; Rajeev, Lara [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division; Ge, Xiaoxuan [Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology; Vaccaro, Brian J. [Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology; Poole, Farris L. [Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology; Arkin, Adam P. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division; Mukhopadhyay, Aindrila [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division; Adams, Michael W. W. [Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology

    2016-12-02

    Cell suspensions of Pelosinus sp. strain UFO1 were previously shown, using spectroscopic analysis, to sequester uranium as U(IV) complexed with carboxyl and phosphoryl group ligands on proteins. The goal of our present study was to characterize the proteins involved in uranium binding. Virtually all of the uranium in UFO1 cells was associated with a heterodimeric protein, which was termed the uranium-binding complex (UBC). The UBC was composed of two S-layer domain proteins encoded by UFO1_4202 and UFO1_4203. Samples of UBC purified from the membrane fraction contained 3.3 U atoms/heterodimer, but significant amounts of phosphate were not detected. The UBC had an estimated molecular mass by gel filtration chromatography of 15 MDa, and it was proposed to contain 150 heterodimers (UFO1_4203 and UFO1_4202) and about 500 uranium atoms. The UBC was also the dominant extracellular protein, but when purified from the growth medium, it contained only 0.3 U atoms/heterodimer. The two genes encoding the UBC were among the most highly expressed genes within the UFO1 genome, and their expressions were unchanged by the presence or absence of uranium. Therefore, the UBC appears to be constitutively expressed and is the first line of defense against uranium, including by secretion into the extracellular medium. Although S-layer proteins were previously shown to bind U(VI), here we showed that U(IV) binds to S-layer proteins, we identified the proteins involved, and we quantitated the amount of uranium bound. Widespread uranium contamination from industrial sources poses hazards to human health and to the environment. Here in this paper, we identified a highly abundant uranium-binding complex (UBC) from Pelosinus sp. strain UFO1. The complex makes up the primary protein component of the S-layer of strain UFO1 and binds 3.3 atoms of U(IV) per heterodimer. Finally, while other bacteria have been shown to bind U(VI) on their S-layer, we demonstrate here an example of U(IV) bound by an S-layer complex. The UBC provides a potential tool for the microbiological sequestration of uranium for the cleaning of contaminated environments.

  20. High-molecular weight star conjugates containing docetaxel with high anti-tumor activity and low systemic toxicity in vivo

    Czech Academy of Sciences Publication Activity Database

    Etrych, Tomáš; Strohalm, Jiří; Šírová, Milada; Tomalová, Barbora; Rossmann, Pavel; Říhová, Blanka; Ulbrich, Karel; Kovář, Marek

    2015-01-01

    Roč. 6, č. 1 (2015), s. 160-170 ISSN 1759-9954 R&D Projects: GA ČR GAP301/11/0325; GA ČR GCP207/12/J030; GA MŠk(CZ) EE2.3.20.0055 Institutional support: RVO:61389013 ; RVO:61388971 Keywords : star polymer * HPMA copolymers * docetaxel Subject RIV: CD - Macromolecular Chemistry ; EC - Immunology (MBU-M) Impact factor: 5.687, year: 2015

  1. Muscular subunits transplantation for facial reanimation

    Directory of Open Access Journals (Sweden)

    Hazan André Salo Buslik

    2006-01-01

    Full Text Available PURPOSE: To present an alternative technique for reconstruction of musculocutaneous damages in the face transferring innervated subsegments(subunits of the latissimus dorsi flap for replacement of various facial mimetic muscles. METHODS: One clinical case of trauma with skin and mimetic muscles damage is described as an example of the technique. The treatment was performed with microsurgical transfer of latissimus dorsi muscle subunits. Each subunit present shape and dimensions of the respective mimetic muscles replaced. The origin, insertions and force vectors for the mimicmuscle lost were considered. Each subsegment has its own arterial and venous supply with a motor nerve component for the muscular unit. RESULTS: Pre and one year postoperative photos registration of static and dynamic mimic aspects, as well as digital electromyography digital data of the patients were compared. The transplanted muscular units presented myoeletric activity, fulfilling both the functional and cosmetic aspect. CONCLUSION: This technique seems to be a promising way to deal with the complex musculocutaneous losses of the face as well as facial palsy.

  2. Influvac, a trivalent inactivated subunit influenza vaccine.

    Science.gov (United States)

    Zuccotti, Gian Vincenzo; Fabiano, Valentina

    2011-01-01

    Influenza represents a major sanitary and socio-economic burden and vaccination is universally considered the most effective strategy for preventing the disease and its complications. Traditional influenza vaccines have been on the market since the late 1940s, with million of doses administered annually worldwide, and demonstrated a substantial efficacy and safety. The trivalent inactivated subunit vaccine has been available for more than 25 years and has been studied in healthy children, adults and the elderly and in people affected by underlying chronic medical conditions. We describe vaccine technology focusing on subunit vaccine production procedures and mode of action and provide updated information on efficacy and safety available data. A review of efficacy and safety data in healthy subjects and in high risk populations from major sponsor- and investigator-driven studies. The vaccine showed a good immunogenicity and a favorable safety profile in all target groups. In the panorama of actually available influenza vaccines, trivalent inactivated subunit vaccine represents a well-established tool for preventing flu and the associated complications.

  3. Journal of Genetics | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Bread-making quality in hexaploid wheats is a complex trait. It has been shown that the amount and composition of protein can influence dough rheological properties. The high-molecular-weight (HMW) glutenins are encoded by a complex locus, Glu-1, on the long arm of group-1 homoeologus chromosome of the A, B and ...

  4. Soybean glycinin subunits: Characterization of physicochemical and adhesion properties.

    Science.gov (United States)

    Mo, Xiaoqun; Zhong, Zhikai; Wang, Donghai; Sun, Xiuzhi

    2006-10-04

    Soybean proteins have shown great potential for applications as renewable and environmentally friendly adhesives. The objective of this work was to study physicochemical and adhesion properties of soy glycinin subunits. Soybean glycinin was extracted from soybean flour and then fractionated into acidic and basic subunits with an estimated purity of 90 and 85%, respectively. Amino acid composition of glycinin subunits was determined. The high hydrophobic amino acid content is a major contributor to the solubility behavior and water resistance of the basic subunits. Acidic subunits and glycinin had similar solubility profiles, showing more than 80% solubility at pH 2.0-4.0 or 6.5-12.0, whereas basic subunits had considerably lower solubility with the minimum at pH 4.5-8.0. Thermal analysis using a differential scanning calorimeter suggested that basic subunits form new oligomeric structures with higher thermal stability than glycinin but no highly ordered structures present in isolated acidic subunits. The wet strength of basic subunits was 160% more than that of acidic subunits prepared at their respective isoelectric points (pI) and cured at 130 degrees C. Both pH and the curing temperature significantly affected adhesive performance. High-adhesion water resistance was usually observed for adhesives from protein prepared at their pI values and cured at elevated temperatures. Basic subunits are responsible for the water resistance of glycinin and are a good starting material for the development of water-resistant adhesives.

  5. Human aldolase B subunit-specific radioimmunoassay

    International Nuclear Information System (INIS)

    Asaka, M.; Alpert, E.

    1983-01-01

    A radioimmunoassay was developed for the direct quantification of aldolase B in human serum and tissues. The method is a double-antibody radioimmunoassay technique using radioiodinated aldolase B homopolymer as ligand, chicken antibodies to aldolase B and rabbit antibodies to chicken IgG. This radioimmunoassay was shown to be specific for the aldolase B subunit, with no cross-reactivity with either human aldolase A subunit or homopolymeric human aldolase C (C 4 ). The lowest measurable amount by this method was 2 ng/ml. Aldolase B is predominantly found in normal liver tissue, with relatively-high aldolase B levels also observed in kidney. Aldolase B levels in the serum obtained from 11 normal subjects ranged from 23 to 38 ng/ml, with a mean of 28.5 +- 9.2 (S.D.) ng/ml. Almost all of patients with hepatitis had serum aldolase B levels greater than 30 ng/ml. In cancer patients, serum aldolase B was slightly elevated in patients with metastatic liver cancer and primary lever cell carcinoma, whereas no elevation of serum aldolase B was shown in patients without liver metastasis. (Auth.)

  6. Subunit stoichiometry of the chloroplast photosystem I complex

    International Nuclear Information System (INIS)

    Bruce, B.D.; Malkin, R.

    1988-01-01

    A native photosystem I (PS I) complex and a PS I core complex depleted of antenna subunits has been isolated from the uniformly 14 C-labeled aquatic higher plant, Lemna. These complexes have been analyzed for their subunit stoichiometry by quantitative sodium dodecyl sulfate-polyacrylamide gel electrophoresis methods. The results for both preparations indicate that one copy of each high molecular mass subunit is present per PS I complex and that a single copy of most low molecular mass subunits is also present. These results suggest that iron-sulfur center X, an early PS I electron acceptor proposed to bind to the high molecular mass subunits, contains a single [4Fe-4S] cluster which is bound to a dimeric structure of high molecular mass subunits, each providing 2 cysteine residues to coordinate this cluster

  7. Transcriptional regulators of Na, K-ATPase subunits

    OpenAIRE

    Zhiqin eLi; Sigrid A Langhans

    2015-01-01

    The Na,K-ATPase classically serves as an ion pump creating an electrochemical gradient across the plasma membrane that is essential for transepithelial transport, nutrient uptake and membrane potential. In addition, Na,K-ATPase also functions as a receptor, a signal transducer and a cell adhesion molecule. With such diverse roles, it is understandable that the Na,K-ATPase subunits, the catalytic alpha-subunit, the beta-subunit and the FXYD proteins, are controlled extensively during developme...

  8. Liposome-Based Adjuvants for Subunit Vaccines: Formulation Strategies for Subunit Antigens and Immunostimulators

    DEFF Research Database (Denmark)

    Schmidt, Signe Tandrup; Foged, Camilla; Korsholm, Karen Smith

    2016-01-01

    be classified into delivery systems or immunostimulators. Liposomes are versatile delivery systems for antigens, and they can carefully be customized towards desired immune profiles by combining them with immunostimulators and optimizing their composition, physicochemical properties and antigen-loading mode......The development of subunit vaccines has become very attractive in recent years due to their superior safety profiles as compared to traditional vaccines based on live attenuated or whole inactivated pathogens, and there is an unmet medical need for improved vaccines and vaccines against pathogens...... of immunostimulators and antigens, respectively, into liposomes, and the choice of immunostimulator should ideally be based on knowledge regarding the specific PRR expression profile of the target APCs. Here, we review state-of-the-art formulation approaches employed for the inclusion of immunostimulators and subunit...

  9. Liposome-Based Adjuvants for Subunit Vaccines: Formulation Strategies for Subunit Antigens and Immunostimulators

    Directory of Open Access Journals (Sweden)

    Signe Tandrup Schmidt

    2016-03-01

    Full Text Available The development of subunit vaccines has become very attractive in recent years due to their superior safety profiles as compared to traditional vaccines based on live attenuated or whole inactivated pathogens, and there is an unmet medical need for improved vaccines and vaccines against pathogens for which no effective vaccines exist. The subunit vaccine technology exploits pathogen subunits as antigens, e.g., recombinant proteins or synthetic peptides, allowing for highly specific immune responses against the pathogens. However, such antigens are usually not sufficiently immunogenic to induce protective immunity, and they are often combined with adjuvants to ensure robust immune responses. Adjuvants are capable of enhancing and/or modulating immune responses by exposing antigens to antigen-presenting cells (APCs concomitantly with conferring immune activation signals. Few adjuvant systems have been licensed for use in human vaccines, and they mainly stimulate humoral immunity. Thus, there is an unmet demand for the development of safe and efficient adjuvant systems that can also stimulate cell-mediated immunity (CMI. Adjuvants constitute a heterogeneous group of compounds, which can broadly be classified into delivery systems or immunostimulators. Liposomes are versatile delivery systems for antigens, and they can carefully be customized towards desired immune profiles by combining them with immunostimulators and optimizing their composition, physicochemical properties and antigen-loading mode. Immunostimulators represent highly diverse classes of molecules, e.g., lipids, nucleic acids, proteins and peptides, and they are ligands for pattern-recognition receptors (PRRs, which are differentially expressed on APC subsets. Different formulation strategies might thus be required for incorporation of immunostimulators and antigens, respectively, into liposomes, and the choice of immunostimulator should ideally be based on knowledge regarding the

  10. Myristoylated α subunits of guanine nucleotide-binding regulatory proteins

    International Nuclear Information System (INIS)

    Buss, J.E.; Mumby, S.M.; Casey, P.J.; Gilman, A.G.; Sefton, B.M.

    1987-01-01

    Antisera directed against specific subunits of guanine nucleotide-binding regulatory proteins (G proteins) were used to immunoprecipitate these polypeptides from metabolically labeled cells. This technique detects, in extracts of a human astrocytoma cell line, the α subunits of G/sub s/ (stimulatory) (α 45 and α 52 ), a 41-kDa subunit of G/sub i/ (inhibitory) (α 41 ), a 40-kDa protein (α 40 ), and the 36-kDa β subunit. No protein that comigrated with the α subunit of G 0 (unknown function) (α 39 ) was detected. In cells grown in the presence of [ 3 H]myristic acid, α 41 and α 40 contained 3 H label, while the β subunit did not. Chemical analysis of lipids attached covalently to purified α 41 and α 39 from bovine brain also revealed myristic acid. Similar analysis of brain G protein β and γ subunits and of G/sub t/ (Transducin) subunits (α, β, and γ) failed to reveal fatty acids. The fatty acid associated with α 41 , α 40 , and α 39 was stable to treatment with base, suggesting that the lipid is linked to the polypeptide via an amide bond. These GTP binding proteins are thus identified as members of a select group of proteins that contains myristic acid covalently attached to the peptide backbone. Myristate may play an important role in stabilizing interactions of G proteins with phospholipid or with membrane-bound proteins

  11. Development of a Subunit Vaccine for Contagious Bovine ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Their work has set the stage for commercial development of a sub-unit vaccine. ... The sub-unit vaccine will be cost-effective, easy to produce, and safe. How it will make a ... IDRC invites applications for the IDRC Doctoral Research Awards.

  12. Studies on the subunits of human glycoprotein hormones in relation to reproduction

    International Nuclear Information System (INIS)

    Hagen, C.

    1977-01-01

    In this review summarising present knowledge of the biological and immunological activity of the subunits of human glycoprotein hormones, the specificity of the α-subunit and β-subunit radioimmunoassays are discussed. The crossreaction studies performed with the α-subunit radioimmunoassays are aummarised in one table while those with the β-subunit radioimmunoassays are presented in a second table. (JIW)

  13. INTRINSIC REGULATION OF HEMOGLOBIN EXPRESSION BY VARIABLE SUBUNIT INTERFACE STRENGTHS

    Science.gov (United States)

    Manning, James M.; Popowicz, Anthony M.; Padovan, Julio C.; Chait, Brian T.; Manning, Lois R.

    2012-01-01

    SUMMARY The expression of the six types of human hemoglobin subunits over time is currently considered to be regulated mainly by transcription factors that bind to upstream control regions of the gene (the “extrinsic” component of regulation). Here we describe how subunit pairing and further assembly to tetramers in the liganded state is influenced by the affinity of subunits for one another (the “intrinsic” component of regulation). The adult hemoglobin dimers have the strongest subunit interfaces and the embryonic hemoglobins are the weakest with fetal hemoglobins of intermediate strength, corresponding to the temporal order of their expression. These variable subunit binding strengths and the attenuating effects of acetylation contribute to the differences with which these hemoglobin types form functional O2-binding tetramers consistent with gene switching. PMID:22129306

  14. Distribution of gluten proteins in bread wheat (Triticum aestivum) grain.

    Science.gov (United States)

    Tosi, Paola; Gritsch, Cristina Sanchis; He, Jibin; Shewry, Peter R

    2011-07-01

    Gluten proteins are the major storage protein fraction in the mature wheat grain. They are restricted to the starchy endosperm, which forms white flour on milling, and interact during grain development to form large polymers which form a continuous proteinaceous network when flour is mixed with water to give dough. This network confers viscosity and elasticity to the dough, enabling the production of leavened products. The starchy endosperm is not a homogeneous tissue and quantitative and qualitative gradients exist for the major components: protein, starch and cell wall polysaccharides. Gradients in protein content and composition are the most evident and are of particular interest because of the major role played by the gluten proteins in determining grain processing quality. Protein gradients in the starchy endosperm were investigated using antibodies for specific gluten protein types for immunolocalization in developing grains and for western blot analysis of protein extracts from flour fractions obtained by sequential abrasion (pearling) to prepare tissue layers. Differential patterns of distribution were found for the high-molecular-weight subunits of glutenin (HMW-GS) and γ-gliadins when compared with the low-molecular-weight subunits of glutenin (LMW-GS), ω- and α-gliadins. The first two types of gluten protein are more abundant in the inner endosperm layers and the latter more abundant in the subaleurone. Immunolocalization also showed that segregation of gluten proteins occurs both between and within protein bodies during protein deposition and may still be retained in the mature grain. Quantitative and qualitative gradients in gluten protein composition are established during grain development. These gradients may be due to the origin of subaleurone cells, which unlike other starchy endosperm cells derive from the re-differentiation of aleurone cells, but could also result from the action of specific regulatory signals produced by the maternal tissue

  15. The light subunit of system bo,+ is fully functional in the absence of the heavy subunit

    OpenAIRE

    Reig, Núria; Chillarón, Josep; Bartoccioni, Paola; Fernández, Esperanza; Bendahan, Annie; Zorzano, Antonio; Kanner, Baruch; Palacín, Manuel; Bertran, Joan

    2002-01-01

    The heteromeric amino acid transporters are composed of a type II glycoprotein and a non-glycosylated polytopic membrane protein. System bo,+ exchanges dibasic for neutral amino acids. It is composed of rBAT and bo,+AT, the latter being the polytopic membrane subunit. Mutations in either of them cause malfunction of the system, leading to cystinuria. bo,+AT-reconstituted systems from HeLa or MDCK cells catalysed transport of arginine that was totally dependent on the presence of one of the bo...

  16. Genetic analysis of the cytoplasmic dynein subunit families.

    Science.gov (United States)

    Pfister, K Kevin; Shah, Paresh R; Hummerich, Holger; Russ, Andreas; Cotton, James; Annuar, Azlina Ahmad; King, Stephen M; Fisher, Elizabeth M C

    2006-01-01

    Cytoplasmic dyneins, the principal microtubule minus-end-directed motor proteins of the cell, are involved in many essential cellular processes. The major form of this enzyme is a complex of at least six protein subunits, and in mammals all but one of the subunits are encoded by at least two genes. Here we review current knowledge concerning the subunits, their interactions, and their functional roles as derived from biochemical and genetic analyses. We also carried out extensive database searches to look for new genes and to clarify anomalies in the databases. Our analysis documents evolutionary relationships among the dynein subunits of mammals and other model organisms, and sheds new light on the role of this diverse group of proteins, highlighting the existence of two cytoplasmic dynein complexes with distinct cellular roles.

  17. Genetic analysis of the cytoplasmic dynein subunit families.

    Directory of Open Access Journals (Sweden)

    K Kevin Pfister

    2006-01-01

    Full Text Available Cytoplasmic dyneins, the principal microtubule minus-end-directed motor proteins of the cell, are involved in many essential cellular processes. The major form of this enzyme is a complex of at least six protein subunits, and in mammals all but one of the subunits are encoded by at least two genes. Here we review current knowledge concerning the subunits, their interactions, and their functional roles as derived from biochemical and genetic analyses. We also carried out extensive database searches to look for new genes and to clarify anomalies in the databases. Our analysis documents evolutionary relationships among the dynein subunits of mammals and other model organisms, and sheds new light on the role of this diverse group of proteins, highlighting the existence of two cytoplasmic dynein complexes with distinct cellular roles.

  18. Transcriptional regulators of Na, K-ATPase subunits

    Directory of Open Access Journals (Sweden)

    Zhiqin eLi

    2015-10-01

    Full Text Available The Na,K-ATPase classically serves as an ion pump creating an electrochemical gradient across the plasma membrane that is essential for transepithelial transport, nutrient uptake and membrane potential. In addition, Na,K-ATPase also functions as a receptor, a signal transducer and a cell adhesion molecule. With such diverse roles, it is understandable that the Na,K-ATPase subunits, the catalytic alpha-subunit, the beta-subunit and the FXYD proteins, are controlled extensively during development and to accommodate physiological needs. The spatial and temporal expression of Na,K-ATPase is partially regulated at the transcriptional level. Numerous transcription factors, hormones, growth factors, lipids and extracellular stimuli modulate the transcription of the Na,K-ATPase subunits. Moreover, epigenetic mechanisms also contribute to the regulation of Na,K-ATPase expression. With the ever growing knowledge about diseases associated with the malfunction of Na,K-ATPase, this review aims at summarizing the best-characterized transcription regulators that modulate Na,K-ATPase subunit levels. As abnormal expression of Na,K-ATPase subunits have been observed in many carcinoma, we will also discuss transcription factors that are associated with epithelial-to-mesenchymal transition, a crucial step in the progression of many tumors to malignant disease.

  19. Pituitary glycoprotein hormone a-subunit secretion by cirrhotic patients

    Directory of Open Access Journals (Sweden)

    Oliveira M.C.

    1999-01-01

    Full Text Available Secretion of the a-subunit of pituitary glycoprotein hormones usually follows the secretion of intact gonadotropins and is increased in gonadal failure and decreased in isolated gonadotropin deficiency. The aim of the present study was to determine the levels of the a-subunit in the serum of patients with cirrhosis of the liver and to compare the results obtained for eugonadal cirrhotic patients with those obtained for cirrhotic patients with hypogonadotropic hypogonadism. Forty-seven of 63 patients with cirrhosis (74.6% presented hypogonadism (which was central in 45 cases and primary in 2, 7 were eugonadal, and 9 women were in normal menopause. The serum a-subunit was measured by the fluorimetric method using monoclonal antibodies. Cross-reactivity with LH, TSH, FSH and hCG was 6.5, 1.2, 4.3 and 1.1%, respectively, with an intra-assay coefficient of variation (CV of less than 5% and an interassay CV of 5%, and sensitivity limit of 4 ng/l. The serum a-subunit concentration ranged from 36 to 6253 ng/l, with a median of 273 ng/l. The median was 251 ng/l for patients with central hypogonadism and 198 ng/l for eugonadal patients. The correlation between the a-subunit and basal LH levels was significant both in the total sample (r = 0.48, P<0.01 and in the cirrhotic patients with central hypogonadism (r = 0.33, P = 0.02. Among men with central hypogonadism there was a negative correlation between a-subunit levels and total testosterone levels (r = 0.54, P<0.01 as well as free testosterone levels (r = -0.53, P<0.01. In conclusion, although the a-subunit levels are correlated with LH levels, at present they cannot be used as markers for hypogonadism in patients with cirrhosis of the liver.

  20. The first transmembrane domain (TM1) of β2-subunit binds to the transmembrane domain S1 of α-subunit in BK potassium channels

    Science.gov (United States)

    Morera, Francisco J.; Alioua, Abderrahmane; Kundu, Pallob; Salazar, Marcelo; Gonzalez, Carlos; Martinez, Agustin D.; Stefani, Enrico; Toro, Ligia; Latorre, Ramon

    2012-01-01

    The BK channel is one of the most broadly expressed ion channels in mammals. In many tissues, the BK channel pore-forming α-subunit is associated to an auxiliary β-subunit that modulates the voltage- and Ca2+-dependent activation of the channel. Structural components present in β-subunits that are important for the physical association with the α-subunit are yet unknown. Here, we show through co-immunoprecipitation that the intracellular C-terminus, the second transmembrane domain (TM2) and the extracellular loop of the β2-subunit are dispensable for association with the α-subunit pointing transmembrane domain 1 (TM1) as responsible for the interaction. Indeed, the TOXCAT assay for transmembrane protein–protein interactions demonstrated for the first time that TM1 of the β2-subunit physically binds to the transmembrane S1 domain of the α-subunit. PMID:22710124

  1. Probing the functional subunits of the tonoplast H+-ATPase

    International Nuclear Information System (INIS)

    Randall, S.K.; Lai, S.; Sze, H.

    1986-01-01

    The tonoplast ATPase of oat roots is composed of at least three polypeptides of 72, 60, and 16 kDa. The 16 kDA polypeptide covalently binds N,N'-dicyclohexylcarbodiimide and is postulated to be a component of the proton channel. Initial studies to identify other subunits indicate that both the 72 and 60 kDa subunits covalently bind 14 C]-7-chloro-4-nitrobenzo-2-oxa-1,3-diazole and [ 14 C]N-ethylamleimide, inhibitors of the tonoplast ATPase. ATP prevents binding of these inhibitors suggesting that both the 72 and 60 kDa subunits are involved in substrate binding. Polyclonal antibody has been made to the 72 kDa subunit. Western blot analysis of tonoplast vesicles reveals single reactive polypeptide (72 kDa). The antibody shows no cross-reactivity towards either the mitochondrial F 1 -ATPase or the plasma membrane ATPase. This antibody specifically inhibits ATP hydrolysis and ATP-dependent H + pumping in native tonoplast vesicles. The authors conclude that the 72 kDa subunit is intimately associated with the catalytic (or ATP-binding) site

  2. Role of regulatory subunits and protein kinase inhibitor (PKI) in determining nuclear localization and activity of the catalytic subunit of protein kinase A.

    Science.gov (United States)

    Wiley, J C; Wailes, L A; Idzerda, R L; McKnight, G S

    1999-03-05

    Regulation of protein kinase A by subcellular localization may be critical to target catalytic subunits to specific substrates. We employed epitope-tagged catalytic subunit to correlate subcellular localization and gene-inducing activity in the presence of regulatory subunit or protein kinase inhibitor (PKI). Transiently expressed catalytic subunit distributed throughout the cell and induced gene expression. Co-expression of regulatory subunit or PKI blocked gene induction and prevented nuclear accumulation. A mutant PKI lacking the nuclear export signal blocked gene induction but not nuclear accumulation, demonstrating that nuclear export is not essential to inhibit gene induction. When the catalytic subunit was targeted to the nucleus with a nuclear localization signal, it was not sequestered in the cytoplasm by regulatory subunit, although its activity was completely inhibited. PKI redistributed the nuclear catalytic subunit to the cytoplasm and blocked gene induction, demonstrating that the nuclear export signal of PKI can override a strong nuclear localization signal. With increasing PKI, the export process appeared to saturate, resulting in the return of catalytic subunit to the nucleus. These results demonstrate that both the regulatory subunit and PKI are able to completely inhibit the gene-inducing activity of the catalytic subunit even when the catalytic subunit is forced to concentrate in the nuclear compartment.

  3. Differential effects of a post-anthesis fertilizer regimen on the wheat flour proteome determined by quantitative 2-DE

    Directory of Open Access Journals (Sweden)

    Altenbach Susan B

    2011-08-01

    Full Text Available Abstract Background Mineral nutrition during wheat grain development has large effects on wheat flour protein content and composition, which in turn affect flour quality and immunogenic potential for a commodity of great economic value. However, it has been difficult to define the precise effects of mineral nutrition on protein composition because of the complexity of the wheat flour proteome. Recent improvements in the identification of flour proteins by tandem mass spectrometry (MS/MS and the availability of a comprehensive proteome map of flour from the US wheat Butte 86 now make it possible to document changes in the proportions of individual flour proteins that result from the application of mineral nutrition. Results Plants of Triticum aestivum 'Butte 86' were grown with or without post-anthesis fertilization (PAF and quantitative 2-dimensional gel electrophoresis (2-DE was used to analyze protein composition of the resulting flour. Significant changes in the proportions of 54 unique proteins were observed as a result of the treatment. Most omega-gliadins, high molecular weight glutenin subunits (HMW-GS and serpins as well as some alpha-gliadins increased in proportion with PAF. In contrast, alpha-amylase/protease inhibitors, farinins, purinins and puroindolines decreased in proportion. Decreases were also observed in several low molecular weight glutenin subunits (LMW-GS, globulins, defense proteins and enzymes. The ratio of HMW-GS to LMW-GS in the flour increased from 0.61 to 0.95 and the ratio of gliadins to glutenins increased from 1.02 to 1.30 with PAF. Because flour protein content doubled with PAF from 7 to 14%, most protein types actually increased in absolute amount (μg/mg flour protein. Data further suggest that flour proteins change with PAF according to their content of sulfur-containing amino acids Cys + Met. Conclusions A 2-DE approach revealed changes in the wheat flour proteome due to PAF that are important for flour

  4. Novel subunit structure observed for noncooperative hemoglobin from Urechis caupo.

    Science.gov (United States)

    Kolatkar, P R; Meador, W E; Stanfield, R L; Hackert, M L

    1988-03-05

    Tetrameric hemoglobin from the "fat innkeeper" worm Urechis caupo possesses a novel subunit arrangement having an "inside out" quaternary structure in that the G/H helices are located on the outer surface of the tetramer. A 5-A resolution crystal structure reveals that although the individual subunits are beta-like, having a distinct D helix and the general myoglobin fold, the subunit contacts are very different from those previously observed for hemoglobins. Furthermore, the hemoglobin from U. caupo is also quite different from the unusual hemoglobin tetramer from clam which also has its G/H helices on the outer surface but with the hemes in close proximity through E-F helical contacts (Royer, W. E., Jr., Love, W. E., and Fenderson, F. F. (1985) Nature 316, 277-280).

  5. Dynamic properties of motor proteins with two subunits

    International Nuclear Information System (INIS)

    Kolomeisky, Anatoly B; III, Hubert Phillips

    2005-01-01

    The dynamics of motor protein molecules consisting of two subunits is investigated using simple discrete stochastic models. Exact steady-state analytical expressions are obtained for velocities and dispersions for any number of intermediate states and conformations between the corresponding binding states of proteins. These models enable us to provide a detailed description and comparison of two different mechanisms of the motion of motor proteins along the linear tracks: the hand-over-hand mechanism, when the motion of subunits alternate; and the inchworm mechanism, when one subunit is always trailing another one. It is shown that the proteins in the hand-over-hand mechanism move faster and fluctuate more than the molecules in the inchworm mechanism. The effect of external forces on dynamic properties of motor proteins is also discussed. Finally, a quantitative method, based on experimental observations for single motor proteins, is proposed for distinguishing between two mechanisms of motion

  6. CSNAP Is a Stoichiometric Subunit of the COP9 Signalosome

    Directory of Open Access Journals (Sweden)

    Shelly Rozen

    2015-10-01

    Full Text Available The highly conserved COP9 signalosome (CSN complex is a key regulator of all cullin-RING-ubiquitin ligases (CRLs, the largest family of E3 ubiquitin ligases. Until now, it was accepted that the CSN is composed of eight canonical components. Here, we report the discovery of an additional integral and stoichiometric subunit that had thus far evaded detection, and we named it CSNAP (CSN acidic protein. We show that CSNAP binds CSN3, CSN5, and CSN6, and its incorporation into the CSN complex is mediated through the C-terminal region involving conserved aromatic residues. Moreover, depletion of this small protein leads to reduced proliferation and a flattened and enlarged morphology. Finally, on the basis of sequence and structural properties shared by both CSNAP and DSS1, a component of the related 19S lid proteasome complex, we propose that CSNAP, the ninth CSN subunit, is the missing paralogous subunit of DSS1.

  7. Cholera Toxin B: One Subunit with Many Pharmaceutical Applications

    Directory of Open Access Journals (Sweden)

    Keegan J. Baldauf

    2015-03-01

    Full Text Available Cholera, a waterborne acute diarrheal disease caused by Vibrio cholerae, remains prevalent in underdeveloped countries and is a serious health threat to those living in unsanitary conditions. The major virulence factor is cholera toxin (CT, which consists of two subunits: the A subunit (CTA and the B subunit (CTB. CTB is a 55 kD homopentameric, non-toxic protein binding to the GM1 ganglioside on mammalian cells with high affinity. Currently, recombinantly produced CTB is used as a component of an internationally licensed oral cholera vaccine, as the protein induces potent humoral immunity that can neutralize CT in the gut. Additionally, recent studies have revealed that CTB administration leads to the induction of anti-inflammatory mechanisms in vivo. This review will cover the potential of CTB as an immunomodulatory and anti-inflammatory agent. We will also summarize various recombinant expression systems available for recombinant CTB bioproduction.

  8. Thermostable Subunit Vaccines for Pulmonary Delivery: How Close Are We?

    DEFF Research Database (Denmark)

    Foged, Camilla

    2016-01-01

    , such as influenza, tuberculosis, and Ebola, for which no good universal vaccines exist. At least two pharmaceutical improvements are expected to help filling this gap: i) The development of thermostable vaccine dosage forms, and ii) the full exploitation of the adjuvant technology for subunit vaccines to potentiate...... strong immune responses. This review highlights the status and recent advances in formulation and pulmonary delivery of thermostable human subunit vaccines. Such vaccines are very appealing from compliance, distribution and immunological point of view: Being non-invasive, inhalable vaccines are self...... immunity. Here, I review state of the art and perspectives in formulation design and processing methods for powder-based subunit vaccines intended for pulmonary administration, and present dry powder inhaler technologies suitable for translating these vaccines into clinical trials....

  9. Catalytic Subunit 1 of Protein Phosphatase 2A Is a Subunit of the STRIPAK Complex and Governs Fungal Sexual Development.

    Science.gov (United States)

    Beier, Anna; Teichert, Ines; Krisp, Christoph; Wolters, Dirk A; Kück, Ulrich

    2016-06-21

    The generation of complex three-dimensional structures is a key developmental step for most eukaryotic organisms. The details of the molecular machinery controlling this step remain to be determined. An excellent model system to study this general process is the generation of three-dimensional fruiting bodies in filamentous fungi like Sordaria macrospora Fruiting body development is controlled by subunits of the highly conserved striatin-interacting phosphatase and kinase (STRIPAK) complex, which has been described in organisms ranging from yeasts to humans. The highly conserved heterotrimeric protein phosphatase PP2A is a subunit of STRIPAK. Here, catalytic subunit 1 of PP2A was functionally characterized. The Δpp2Ac1 strain is sterile, unable to undergo hyphal fusion, and devoid of ascogonial septation. Further, PP2Ac1, together with STRIPAK subunit PRO22, governs vegetative and stress-related growth. We revealed in vitro catalytic activity of wild-type PP2Ac1, and our in vivo analysis showed that inactive PP2Ac1 blocks the complementation of the sterile deletion strain. Tandem affinity purification, followed by mass spectrometry and yeast two-hybrid analysis, verified that PP2Ac1 is a subunit of STRIPAK. Further, these data indicate links between the STRIPAK complex and other developmental signaling pathways, implying the presence of a large interconnected signaling network that controls eukaryotic developmental processes. The insights gained in our study can be transferred to higher eukaryotes and will be important for understanding eukaryotic cellular development in general. The striatin-interacting phosphatase and kinase (STRIPAK) complex is highly conserved from yeasts to humans and is an important regulator of numerous eukaryotic developmental processes, such as cellular signaling and cell development. Although functional insights into the STRIPAK complex are accumulating, the detailed molecular mechanisms of single subunits are only partially understood

  10. Evaluation of subunit vaccines against feline immunodeficiency virus infection

    NARCIS (Netherlands)

    Horzinek, M.C.; Verschoor, E.J.; Willemse, M.J.; Stam, J.G.; Vliet, A.L.W. van; Pouwels, H.; Chalmers, S.K.; Sondermeijer, P.J.; Hesselink, W.; Ronde, A. de

    1996-01-01

    Subunit vaccines prepared against feline immunodeficiency virus (FIV) infection were evaluated in two trials. First, cats were immunized with bacterial expression products of an envelope fragment that contained the V3 neutralization domain of the FIV surface protein fused to either galactokinase

  11. Partial agonists and subunit selectivity at NMDA receptors

    DEFF Research Database (Denmark)

    Risgaard, Rune; Hansen, Kasper Bø; Clausen, Rasmus Prætorius

    2010-01-01

    Subunit-selective ligands for glutamate receptors remains an area of interest as glutamate is the major excitatory neurotransmitter in the brain and involved in a number of diseased states in the central nervous system (CNS). Few subtype-selective ligands are known, especially among the N...

  12. Therapeutic potential of Mediator complex subunits in metabolic diseases.

    Science.gov (United States)

    Ranjan, Amol; Ansari, Suraiya A

    2018-01-01

    The multisubunit Mediator is an evolutionary conserved transcriptional coregulatory complex in eukaryotes. It is needed for the transcriptional regulation of gene expression in general as well as in a gene specific manner. Mediator complex subunits interact with different transcription factors as well as components of RNA Pol II transcription initiation complex and in doing so act as a bridge between gene specific transcription factors and general Pol II transcription machinery. Specific interaction of various Mediator subunits with nuclear receptors (NRs) and other transcription factors involved in metabolism has been reported in different studies. Evidences indicate that ligand-activated NRs recruit Mediator complex for RNA Pol II-dependent gene transcription. These NRs have been explored as therapeutic targets in different metabolic diseases; however, they show side-effects as targets due to their overlapping involvement in different signaling pathways. Here we discuss the interaction of various Mediator subunits with transcription factors involved in metabolism and whether specific interaction of these transcription factors with Mediator subunits could be potentially utilized as therapeutic strategy in a variety of metabolic diseases. Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

  13. α-4 subunit of nicotinic acetylcholine receptor polymorphisms exhibit ...

    African Journals Online (AJOL)

    Background: Smoking behavior is influenced by both genetic and environmental factors. Nicotine is the major addictive substance in cigarettes. Nicotinic acetylcholine receptors (nAChRs) are thought to play an important role in nicotine addiction of smokers. One of the genes, α-4 subunit of nicotinic acetylcholine receptor ...

  14. Structural interaction of novel dendrimer and subunits with water

    African Journals Online (AJOL)

    Preferred Customer

    interaction study with solvents are essential [4-6] and several subunits are used for .... slowed down the viscous flow with higher excess limiting viscosities of the 2,4,6- ..... Practical Organic Chemistry, 5th ed.; Wiley: New York; 1989; p 300. 14.

  15. Bacterial cellulose biosynthesis: diversity of operons, subunits, products and functions

    Science.gov (United States)

    Römling, Ute; Galperin, Michael Y.

    2015-01-01

    Summary Recent studies of bacterial cellulose biosynthesis, including structural characterization of a functional cellulose synthase complex, provided the first mechanistic insight into this fascinating process. In most studied bacteria, just two subunits, BcsA and BcsB, are necessary and sufficient for the formation of the polysaccharide chain in vitro. Other subunits – which differ among various taxa – affect the enzymatic activity and product yield in vivo by modulating expression of biosynthesis apparatus, export of the nascent β-D-glucan polymer to the cell surface, and the organization of cellulose fibers into a higher-order structure. These auxiliary subunits play key roles in determining the quantity and structure of the resulting biofilm, which is particularly important for interactions of bacteria with higher organisms that lead to rhizosphere colonization and modulate virulence of cellulose-producing bacterial pathogens inside and outside of host cells. Here we review the organization of four principal types of cellulose synthase operons found in various bacterial genomes, identify additional bcs genes that encode likely components of the cellulose biosynthesis and secretion machinery, and propose a unified nomenclature for these genes and subunits. We also discuss the role of cellulose as a key component of biofilms formed by a variety of free-living and pathogenic bacteria and, for the latter, in the choice between acute infection and persistence in the host. PMID:26077867

  16. Moessbauer spectroscopic studies of hemoglobin and its isolated subunits

    International Nuclear Information System (INIS)

    Hoy, G.R.; Cook, D.C.; Berger, R.L.; Friedman, F.K.

    1986-01-01

    Samples of 90% enriched 57Fe hemoglobin and its isolated subunits have been prepared. Moessbauer spectroscopic measurements have been made on three such samples. Sample one contained contributions of oxyhemoglobin, deoxyhemoglobin, and carbonmonoxyhemoglobin. This sample was studied from a temperature of 90 K down to 230 mK. Measurements were also made at 4.2 K using a small applied magnetic field of 1.0 T. In general, the measured quadrupole splittings and isomer shifts for each component agreed with previous measurements on single component samples in the literature, and thus demonstrated that chemically enriched hemoglobin has not been altered. The second and third samples were isolated alpha and beta subunits, respectively. We have found measurable Moessbauer spectral differences between the HbO 2 sites in the alpha subunit sample and the beta subunit sample. The measured Moessbauer spectral areas indicate that the iron ion has the largest mean-square displacement at the deoxy Hb sites as compared to that at the oxy- and carbonmonoxy Hb sites. The mean-square displacement at the HbO 2 sites is the smallest

  17. Characterization of an Integrated Active Glu-1Ay Allele in Common Wheat from Wild Emmer and Its Potential Role in Flour Improvement

    Directory of Open Access Journals (Sweden)

    Zhenzhen Wang

    2018-03-01

    Full Text Available Glu-1Ay, one of six genes encoding a high molecular weight glutenin subunit (HMW-GS, is frequently silenced in hexaploid common wheat. Here, an active allele of Glu-1Ay was integrated from wild emmer wheat (Triticum turgidum ssp. dicoccoides accession D97 into the common wheat (Triticum aestivum cultivar Chuannong 16 via the repeated self-fertilization of the pentaploid interspecific hybrid, culminating in the selection of a line TaAy7-40 shown to express the wild emmer Glu-1Ay allele. The open reading frame of this allele was a 1830 bp long sequence, demonstrated by its heterologous expression in Escherichia coli to encode a 608-residue polypeptide. Its nucleotide sequence was 99.2% identical to that of the sequence within the wild emmer parent. The TaAy7-40 introgression line containing the active Glu-1Ay allele showed higher protein content, higher sodium dodecyl sulfate (SDS sedimentation value, higher content of wet gluten in the flour, higher grain weight, and bigger grain size than Chuannong 16. The end-use quality parameters of the TaAy7-40 were superior to those of the medium gluten common wheat cultivars Mianmai 37 and Neimai 9. Thus, the active Glu-1Ay allele might be of potential value in breeding programs designed to improve wheat flour quality.

  18. Genetic diversity, population structure and marker-trait associations for agronomic and grain traits in wild diploid wheat Triticum urartu.

    Science.gov (United States)

    Wang, Xin; Luo, Guangbin; Yang, Wenlong; Li, Yiwen; Sun, Jiazhu; Zhan, Kehui; Liu, Dongcheng; Zhang, Aimin

    2017-07-01

    Wild diploid wheat, Triticum urartu (T. urartu) is the progenitor of bread wheat, and understanding its genetic diversity and genome function will provide considerable reference for dissecting genomic information of common wheat. In this study, we investigated the morphological and genetic diversity and population structure of 238 T. urartu accessions collected from different geographic regions. This collection had 19.37 alleles per SSR locus and its polymorphic information content (PIC) value was 0.76, and the PIC and Nei's gene diversity (GD) of high-molecular-weight glutenin subunits (HMW-GSs) were 0.86 and 0.88, respectively. UPGMA clustering analysis indicated that the 238 T. urartu accessions could be classified into two subpopulations, of which Cluster I contained accessions from Eastern Mediterranean coast and those from Mesopotamia and Transcaucasia belonged to Cluster II. The wide range of genetic diversity along with the manageable number of accessions makes it one of the best collections for mining valuable genes based on marker-trait association. Significant associations were observed between simple sequence repeats (SSR) or HMW-GSs and six morphological traits: heading date (HD), plant height (PH), spike length (SPL), spikelet number per spike (SPLN), tiller angle (TA) and grain length (GL). Our data demonstrated that SSRs and HMW-GSs were useful markers for identification of beneficial genes controlling important traits in T. urartu, and subsequently for their conservation and future utilization, which may be useful for genetic improvement of the cultivated hexaploid wheat.

  19. Assessment of the Allergenic Potential of Transgenic Wheat (Triticum aestivum) with Reduced Levels of ω5-Gliadins, the Major Sensitizing Allergen in Wheat-Dependent Exercise-Induced Anaphylaxis.

    Science.gov (United States)

    Altenbach, Susan B; Tanaka, Charlene K; Pineau, Florence; Lupi, Roberta; Drouet, Martine; Beaudouin, Etienne; Morisset, Martine; Denery-Papini, Sandra

    2015-10-28

    The ω5-gliadins are the major sensitizing allergens in wheat-dependent exercise-induced anaphylaxis (WDEIA). In this study, two-dimensional immunoblot analysis was used to assess the allergenic potential of two transgenic wheat lines in which ω5-gliadin genes were silenced by RNA interference. Sera from 7 of 11 WDEIA patients showed greatly reduced levels of immunoglobulin E (IgE) reactivity to ω5-gliadins in both transgenic lines. However, these sera also showed low levels of reactivity to other gluten proteins. Sera from three patients showed the greatest reactivity to proteins other than ω5-gliadins, either high-molecular-weight glutenin subunits (HMW-GSs), α-gliadins, or non-gluten proteins. The complexity of immunological responses among these patients suggests that flour from the transgenic lines would not be suitable for individuals already diagnosed with WDEIA. However, the introduction of wheat lacking ω5-gliadins could reduce the number of people sensitized to these proteins and thereby decrease the overall incidence of this serious food allergy.

  20. Protein-transitions in and out of the dough matrix in wheat flour mixing.

    Science.gov (United States)

    Wang, Xiaolong; Appels, Rudi; Zhang, Xiaoke; Bekes, Ferenc; Torok, Kitti; Tomoskozi, Sandor; Diepeveen, Dean; Ma, Wujun; Islam, Shahidul

    2017-02-15

    Sequential protein behavior in the wheat dough matrix under continuous mixing and heating treatment has been studied using Mixolab-dough samples from two Australian wheat cultivars, Westonia and Wyalkatchem. Size exclusion high performance liquid chromatography (SE-HPLC) and two-dimensional gel electrophoresis (2-DGE) analysis indicated that 32min (80°C) was a critical time point in forming large protein complexes and loosing extractability of several protein groups like y-type high molecular weight glutenin subunits (HMW-GSs), gamma-gliadins, beta-amylases, serpins, and metabolic proteins with higher mass. Up to 32min (80°C) Westonia showed higher protein extractability compared to Wyalkatchem although it was in the opposite direction thereafter. Twenty differentially expressed proteins could be assigned to chromosomes 1D, 3A, 4A, 4B, 4D, 6A, 6B, 7A and 7B. The results expanded the range of proteins associated with changes in the gluten-complex during processing and provided targets for selecting new genetic variants associated with altered quality attributes of the flour. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Development of a peptide substrate for detection of Sunn pest damage in wheat flour.

    Science.gov (United States)

    Hançerlioğulları, Begüm Zeynep; Köksel, Hamit; Dudak, Fahriye Ceyda

    2018-05-07

    Since the common protease substrates did not give satisfactory results for the determination of Sunn pest protease activity in damaged wheat, different peptide substrates derived from the repeat sequences of high molecular weight glutenin subunits were synthesized. Hydrolysis of peptides by pest protease was determined by HPLC. Among three peptides having the same consensus motifs, peptide1 (PGQGQQGYYPTSPQQ) showed the best catalytic efficiency. A novel assay was described for monitoring the enzymatic activity of protease extracted from damaged wheat flour. The selected peptide was labeled with a fluorophore (EDANS) and quencher (Dabcyl) to display fluorescence resonance energy transfer (FRET). The proteolytic activity was measured by the change in fluorescence intensity that occurred when the protease cleaved the peptide substrate. Furthermore, the developed assay was modified for rapid and easy detection of bug damage in flour. Flour samples were suspended in water and mixed with fluorescence peptide substrate. After centrifugation, the fluorescence intensities of the supernatants were determined which is proportional with the protease content of the flour. The total analysis time for the developed assay is estimated as 15 minutes. The developed assay permits a significant decrease in time and labor, offering sensitive detection of Sunn pest damage in wheat flour. This article is protected by copyright. All rights reserved.

  2. 100 Gy 60Co γ-Ray Induced Novel Mutations in Tetraploid Wheat

    Directory of Open Access Journals (Sweden)

    Chuntao Yang

    2014-01-01

    Full Text Available 10 accessions of tetraploid wheat were radiated with 100 Gy 60Co γ-ray. The germination energy, germination rate, special characters (secondary tillering, stalk with wax powder, and dwarf, meiotic process, and high-molecular-weight glutenin subunits (HMW-GSs were observed. Different species has different radiation sensibility. With 1 seed germinated (5%, T. dicoccum (PI434999 is the most sensitive to this dose of radiation. With a seed germination rate of 35% and 40%, this dose also affected T. polonicum (As304 and T. carthlicum (As293. Two mutant dwarf plants, T. turgidum (As2255 253-10 and T. polonicum (As302 224-14, were detected. Abnormal chromosome pairings were observed in pollen mother cells of both T. dicoccoides (As835 237-9 and T. dicoccoides (As838 239-8 with HMW-GS 1Ax silent in seeds from them. Compared with the unirradiated seed of T. polonicum (As304 CK, a novel HMW-GS was detected in seed of T. polonicum (As304 230-7 and its electrophoretic mobility was between 1By8 and 1Dy12 which were the HMW-GSs of Chinese Spring. These mutant materials would be resources for wheat breeding.

  3. Catalytic Subunit 1 of Protein Phosphatase 2A Is a Subunit of the STRIPAK Complex and Governs Fungal Sexual Development

    Directory of Open Access Journals (Sweden)

    Anna Beier

    2016-06-01

    Full Text Available The generation of complex three-dimensional structures is a key developmental step for most eukaryotic organisms. The details of the molecular machinery controlling this step remain to be determined. An excellent model system to study this general process is the generation of three-dimensional fruiting bodies in filamentous fungi like Sordaria macrospora. Fruiting body development is controlled by subunits of the highly conserved striatin-interacting phosphatase and kinase (STRIPAK complex, which has been described in organisms ranging from yeasts to humans. The highly conserved heterotrimeric protein phosphatase PP2A is a subunit of STRIPAK. Here, catalytic subunit 1 of PP2A was functionally characterized. The Δpp2Ac1 strain is sterile, unable to undergo hyphal fusion, and devoid of ascogonial septation. Further, PP2Ac1, together with STRIPAK subunit PRO22, governs vegetative and stress-related growth. We revealed in vitro catalytic activity of wild-type PP2Ac1, and our in vivo analysis showed that inactive PP2Ac1 blocks the complementation of the sterile deletion strain. Tandem affinity purification, followed by mass spectrometry and yeast two-hybrid analysis, verified that PP2Ac1 is a subunit of STRIPAK. Further, these data indicate links between the STRIPAK complex and other developmental signaling pathways, implying the presence of a large interconnected signaling network that controls eukaryotic developmental processes. The insights gained in our study can be transferred to higher eukaryotes and will be important for understanding eukaryotic cellular development in general.

  4. Metal-​based catalysts for controlled ring-​opening polymerization of macrolactones : high molecular weight and well-​defined copolymer architectures

    NARCIS (Netherlands)

    Bouyahyi, M.; Duchateau, R.

    2014-01-01

    This contribution describes our recent results regarding the metal-¿catalyzed ring-¿opening polymn. of pentadecalactone and its copolymn. with e-¿caprolactone involving single-¿site metal complexes based on aluminum, zinc, and calcium. Under the right conditions (i.e., monomer concn., catalyst type,

  5. High molecular weight glucan of the culinary medicinal mushroom Agaricus bisporus is an a-glucan that forms complexes with low molecular weight galactan

    NARCIS (Netherlands)

    Smiderle, F.; Sassaki, G.L.; Arkel, van J.; Lacomini, M.; Wichers, H.J.; Griensven, van L.J.L.D.

    2010-01-01

    An a-glucan was isolated from the culinary medicinal mushroom A. bisporus by hot water extraction, ethanol precipitation and DEAE-cellulose chromatography. The resulting material showed a single HMW peak excluded from a Sephadex G50 column that could completely be degraded by a-amylase treatment.

  6. The effect of radiation dose on the crosslink density of ultra-high molecular weight polyethylene (UHMWPE) measured by a novel swelling method

    International Nuclear Information System (INIS)

    Muratoglu, O.K.; Bragdon, C.R.; O'Connor, D.O.; Jasty, M.; Harris, W.H.

    1998-01-01

    The crosslink density of a polyethylene network structure can be determined by swelling in hot xylene (130 deg C). The Flory's swelling theory is generally used to calculate the crosslink density, dx (ln(l-q -1 ) + q -1 + Xq -1 )/(V l q -1/3 ), where V l is the molar volume of xylene at 130 deg C (136 cc/mol), X is the xylene-polyethylene interaction parameter, and q is the equilibrium volume swelling ratio of cross-linked network in hot xylene. Conventionally, q is measured using gravimetric methods as described in ASTM D2765-95. However, as noted in the ASTM standard, the gravimetric method has a large error factor associated with the measurement of q (as much as 100%). UHMWPE was irradiated (range of 25 to 300 kGy) using an AECL I 10/1 linear electron beam accelerator operated at 1 kW. The irradiated specimens were subsequently melt-annealed at 150 deg C for 2 hours in vacuum. For swelling experiments, 2 mm thin samples were machined using a diamond blade. The sample sizes were kept at around 3x3x2 mm and the bottom and top surfaces were machined parallel to each other. The equilibrium volume swelling ratios were determined using a Perkin-Elmer TMA/DMA 7 (n=3 for each radiation dose level). The samples were placed in a quartz basket-probe assembly and lowered into a xylene/antioxidant bath at room temperature. The xylene was then heated to 130 deg C at 5 deg C/min and held at 130 deg C for 2 hours. The swelling was then recorded with the upward motion of the probe until the equilibrium swelling was achieved. (The experiments were carried out in 3 orthogonal directions which confirmed the isotropy of swelling). From this one-dimensional change in height, q was calculated by taking into account the volumetric expansion due to heating and melting. (author)

  7. The effect of radiation dose on the crosslink density of ultra-high molecular weight polyethylene (UHMWPE) measured by a novel swelling method

    International Nuclear Information System (INIS)

    Muratoglu, O.K.; Bragdon, C.R.; O'Connor, D.O.; Jasty, M.; Harris, W.H.

    1998-01-01

    The crosslink density of a polyethylene network structure can be determined by swelling in hot xylene (130 deg C). The Flory's swelling theory is generally used to calculate the crosslink density, dx (ln(l-q -1 )+q -1 +Xq -1 )/(V 1 q -1/3 ), where V 1 is the molar volume of xylene at 130 deg C (136 cc/mol), X is the xylene-polyethylene interaction parameter, and q is the equilibrium volume swelling ratio of cross-linked network in hot xylene. Conventionally, q is measured using gravimetric methods as described in ASTM D2765-95. However, as noted in the ASTM standard, the gravimetric method has a large error factor associated with the measurement of q (as much as 100%). UHMWPE was irradiated (range of 25 to 300 kGy) using an AECL I 10/1 linear electron beam accelerator operated at 1 kW. The irradiated specimens were subsequently melt-annealed at 150 deg C for 2 hours in vacuum. For swelling experiments, 2 mm thin samples were machined using a diamond blade. The sample sizes were kept at around 3x3x2 mm and the bottom and top surfaces were machined parallel to each other. The equilibrium volume swelling ratios were determined using a Perkin-Elmer TMA/DMA 7 (n=3 for each radiation dose level). The samples were placed in a quartz basket-probe assembly and lowered into a xylene/antioxidant bath at room temperature. The xylene was then heated to 130 deg C at 5 deg C/min and held at 130 deg C for 2 hours. The swelling was then recorded with the upward motion of the probe until the equilibrium swelling was achieved. (The experiments were carried out in 3 orthogonal directions which confirmed the isotropy of swelling). (author)

  8. Detection of a high-molecular-weight LHRH precursor by cell-free translation of mRNA from human, rat, and mouse hypothalamus

    International Nuclear Information System (INIS)

    Curtis, A.; Szelke, M.; Fink, G.

    1986-01-01

    Large precursors have also been predicted using the immunoprecipitation technique which relies upon the identification of large immunoreactive molecules following in vitro translation of mRNA. The mRNA is presumed to represent the largest form of a nascent precursor polypeptide molecule irrespective of the number of biosynthetic cleavage steps which are necessary to liberate the active peptide. However, as has been shown for somatostatin, nonprotein modifications may be made which apparently increase molecular weight, such as glycosylation or phosphorylation of the molecule. The authors employed the immunoprecipitation technique to confirm earlier chromatographic studies that the hypothalamic decapeptide, luteinizing hormone releasing hormone (LHRH) is also synthesized by way of a large precursor form. The authors' finding show that the translation of hypothalamic mRNA produces a primary translation product with an apparent molecule weight of 28,000 which contains an amino acid sequence immunologically similar to that of biologically active LHRH. The procedure involved the incorporation of a radioactive amino acid into polypeptides synthesized by in vitro translation of hypothalamic messenger RNA. The resulting complex protein mixture was immunoprecipitated with a specific anti-LHRH serum, and the immunoprecipitate was identified by polyacrylamide gel electrophoresis and autoradiography

  9. High-Performance n-Channel Organic Transistors Using High-Molecular-Weight Electron-Deficient Copolymers and Amine-Tailed Self-Assembled Monolayers.

    Science.gov (United States)

    Wang, Yang; Hasegawa, Tsukasa; Matsumoto, Hidetoshi; Mori, Takehiko; Michinobu, Tsuyoshi

    2018-03-01

    While high-performance p-type semiconducting polymers are widely reported, their n-type counterparts are still rare in terms of quantity and quality. Here, an improved Stille polymerization protocol using chlorobenzene as the solvent and palladium(0)/copper(I) as the catalyst is developed to synthesize high-quality n-type polymers with number-average molecular weight up to 10 5 g mol -1 . Furthermore, by sp 2 -nitrogen atoms (sp 2 -N) substitution, three new n-type polymers, namely, pBTTz, pPPT, and pSNT, are synthesized, and the effect of different sp 2 -N substitution positions on the device performances is studied for the first time. It is found that the incorporation of sp 2 -N into the acceptor units rather than the donor units results in superior crystalline microstructures and higher electron mobilities. Furthermore, an amine-tailed self-assembled monolayer (SAM) is smoothly formed on a Si/SiO 2 substrate by a simple spin-coating technique, which can facilitate the accumulation of electrons and lead to more perfect unipolar n-type transistor performances. Therefore, a remarkably high unipolar electron mobility up to 5.35 cm 2 V -1 s -1 with a low threshold voltage (≈1 V) and high on/off current ratio of ≈10 7 is demonstrated for the pSNT-based devices, which are among the highest values for unipolar n-type semiconducting polymers. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Inflammatory Adipokines Decrease Expression of Two High Molecular Weight Isoforms of Tropomyosin Similar to the Change in Type 2 Diabetic Patients.

    Directory of Open Access Journals (Sweden)

    Stuart A Savill

    Full Text Available Cardiovascular disease and cancer are increased in Type 2 diabetes. TPM1 and TPM4 genes encode proteins associated with cardiovascular and neoplastic disease. High (HMW and low (LMW molecular weight isoforms from TPM1 and TPM4 are altered in several cancer cells and the 3'UTR of TPM1 mRNA is tumour suppressive. Leukocytes influence cardiovascular and neoplastic disease by immunosurveillance for cancer and by chronic inflammation in Type 2 diabetes and cardiovascular disease. The aim was to determine changes in expression of isoforms from TPM1 and TPM4 genes in leukocytes from Type 2 diabetic patients and to use the leukocyte cell line THP1 to identify possible mediators of changes in the patients. Gene expression was determined by RT-qPCR. In diabetes, expression of HMW isoforms from TPM1 were markedly decreased (0.55 v 1.00; p = 0.019 but HMW isoforms from TPM4 were not significantly different (0.76 v 1.00; p = 0.205. Within individual variance in expression of HMW isoforms was very high. The change in expression in HMW isoforms from TPM1 and TPM4 was replicated in THP1 cells treated with 1 ng/ml TNFα (0.10 and 0.12 v 1.00 respectively or 10 ng/ml IL-1α (0.17 and 0.14 v 1.00 respectively. Increased insulin or glucose concentrations had no substantial effects on TPM1 or TPM4 expression. Decreased TPM1 mRNA resulted in decreases in HMW protein levels. Expression of HMW isoforms from TPM1 is decreased in Type 2 diabetes. This is probably due to increased levels of inflammatory cytokines TNFα and IL-1α in Type 2 diabetes. Lower levels of TPM1 mRNA reduce tumour suppression and could contribute to increased cancer risk in Type 2 diabetes. Decreased HMW tropomyosin isoforms are associated with cancer. Decreased HMW isoforms give rise to cells that are more plastic, motile, invasive and prone to dedifferentiation resulting in leukocytes that are more invasive but less functionally effective.

  11. Overexpression of high molecular weight FGF-2 forms inhibits glioma growth by acting on cell-cycle progression and protein translation

    International Nuclear Information System (INIS)

    Lemiere, Sylvie; Azar, Rania; Belloc, Francis; Guersel, Demir; Pyronnet, Stephane; Bikfalvi, Andreas; Auguste, Patrick

    2008-01-01

    In order to clarify the role of HMW FGF-2 in glioma development and angiogenesis, we over-expressed different human FGF-2 isoforms in C6 rat glioma cell line using a tetracycline-regulated expression system. Phenotypic modifications were analyzed in vitro and compared to untransfected cells or to cells over-expressing 18 kDa FGF-2 or all FGF-2 isoforms. In particular, we demonstrate that HMW FGF-2 has unique features in inhibiting glioma cell proliferation. HMW FGF-2 expressing cells showed a cell-cycle arrest at the G2M, demonstrating a role of HMW FGF-2 in controlling the entry in mitosis. Moreover, hydroxyurea was ineffective in blocking cells at the G1S boundary when HMW FGF-2 was expressed. We also show that the HMW FGF-2 isoforms inhibit 4E-BP1 phosphorylation at critical sites restoring the translation inhibitory activity of 4E-BP1. In vivo, inhibition of tumor growth was observed when cells expressed HMW FGF-2. This indicates that HMW FGF-2 inhibits tumor growth in glioma cells by acting on cell-cycle progression and protein translation

  12. Affinity chromatographic purification of tetrodotoxin by use of tetrodotoxin-binding high molecular weight substances in the body fluid of shore crab (Hemigrapsus sanguineus) as ligands.

    Science.gov (United States)

    Shiomi, K; Yamaguchi, S; Shimakura, K; Nagashima, Y; Yamamori, K; Matsui, T

    1993-12-01

    A purification method for tetrodotoxin (TTX), based on affinity chromatography using the TTX-binding high mol. wt substances in the body fluid of shore crab (Hemigrapsus sanguineus) as ligands, was developed. This method was particularly useful for analysis of TTX in biological samples with low concentrations of TTX. The affinity gel prepared was highly specific for TTX, having no ability to bind 4-epi-TTX and anhydro-TTX as well as saxitoxin.

  13. Reduced and high molecular weight barley beta-glucans decrease plasma total and non-HDL-cholesterol in hypercholesterolemic Syrian golden hamsters.

    Science.gov (United States)

    Wilson, Thomas A; Nicolosi, Robert J; Delaney, Bryan; Chadwell, Kim; Moolchandani, Vikas; Kotyla, Timothy; Ponduru, Sridevi; Zheng, Guo-Hua; Hess, Richard; Knutson, Nathan; Curry, Leslie; Kolberg, Lore; Goulson, Melanie; Ostergren, Karen

    2004-10-01

    Consumption of concentrated barley beta-glucan lowers plasma cholesterol because of its soluble dietary fiber nature. The role of molecular weight (MW) in lowering serum cholesterol is not well established. Prior studies showed that enzymatic degradation of beta-glucan eliminates the cholesterol-lowering activity; however, these studies did not evaluate the MW of the beta-glucan. The current study was conducted to evaluate whether barley beta-glucan concentrates, partially hydrolyzed to reduce MW, possess cholesterol-lowering and antiatherogenic activities. The reduced MW fraction was compared with a high MW beta-glucan concentrate from the same barley flour. Concentrated beta-glucan preparations were evaluated in Syrian Golden F(1)B hamsters fed a hypercholesterolemic diet (HCD) with cholesterol, hydrogenated coconut oil, and cellulose. After 2 wk, hamsters were fed HCD or diets that contained high or reduced MW beta-glucan at a concentration of 8 g/100 g at the expense of cellulose. Decreases in plasma total cholesterol (TC) and non-HDL-cholesterol (non-HDL-C) concentrations occurred in the hamsters fed reduced MW and high MW beta-glucan diets. Plasma HDL-C concentrations did not differ. HCD-fed hamsters had higher plasma triglyceride concentrations. Liver TC, free cholesterol, and cholesterol ester concentrations did not differ. Aortic cholesterol ester concentrations were lower in the reduced MW beta-glucan-fed hamsters. Consumption of either high or reduced MW beta-glucan increased concentrations of fecal total neutral sterols and coprostanol, a cholesterol derivative. Fecal excretion of cholesterol was greater than in HCD-fed hamsters only in those fed the reduced MW beta-glucan. Study results demonstrate that the cholesterol-lowering activity of barley beta-glucan may occur at both lower and higher MW.

  14. Gelation of Photonic Microdomain Structures Formed in Semi-Dilute Solutions of Ultra-High-Molecular-Weight Polystyrene-b-Polybutadiene with Various Polybutadiene Contents

    International Nuclear Information System (INIS)

    Okamoto, S; Ito, S; Ando, K; Mouri, M; Ikeda, A; Hasegawa, H; Koshikawa, N

    2010-01-01

    Well-ordered microdomain structures were obtained in semi-dilute solutions and successfully stabilized by gelation. We used polystyrene-b-polybutadiene (PS-b-PB) diblock copolymer with the weight-averaged molecular weight varying from several hundred thousands to millions g/mol. The solutions had iridescent colors because the domain spacing is on the order of the wavelength of visible light. As the structures are susceptible to distortion by flow or vibration, structural fixation was carried out by gelation. The polybutadiene used has the microstructure of 1,2-linkage and hence the chains can be cross-linkable. The Small-Angle X-ray Scattering and the UV-vis spectroscopic measurements showed that in the case of PS-b-PBs with the PB volume fraction, φ PB , greater than about 50 vol % the microdomain structures were successfully fixed by gelation, while largely distorted in the case of those with φ PB < ca. 50 vol %. The SAXS scattering intensities were quantitatively analyzed by the scattering functions numerically calculated based on the one- and two-dimensional paracrystal theories and on the concentration fluctuation between the polymers and the solvent molecules.

  15. Occurrence of high molecular weight lipids (C{sub 80+}) in the trilaminar outer cell walls of some freshwater microalgae. A reappraisal of algaenan structure

    Energy Technology Data Exchange (ETDEWEB)

    Allard, B.; Templier, J. [UMR CNRS, Paris (France). Laboratoire de Chimie Bioorganique et Organique Physique; Rager, M.-N. [UMR CNRS, Paris (France). Service RMN

    2002-07-01

    The purified cell walls of mother cells (CWM) were isolated from three strains of trilaminar sheath (TLS)- and algaenan-containing freshwater microalgae Chlorella emersonii, Tetraedron minimum and Scenedesmus communis. The chemical structures of CWM and algaenans were investigated by means of tetramethylammonium hydroxide (TMAH) hydrolysis and tetramethylammonium hydroxide thermochemolysis. The compounds released were characterised by {sup 1}H and {sup 13}C-NMR, gel permeation chromatography and desorption chemical ionisation mass spectrometry. The results show that the outer cell walls of the microalgae are constituted, at least in part, of linear (poly)esters containing extremely long chain alcohol and acid moieties (up to C{sub 80}) and that algaenans are mainly composed of extremely long chain (di)carboxylic acids up to C{sub 120}. The present results which are in direct contrast to the previous three-dimensional architecture proposed for algaenans, led us to re-interpret the algaenan structure. (Author)

  16. Free-standing thermo-responsive nanoporous membranes from high molecular weight PS-PNIPAM block copolymers synthesized via RAFT polymerization

    NARCIS (Netherlands)

    Cetintas, Merve; de Grooth, Joris; Hofman, Anton H.; van der Kooij, Hanne M.; Loos, Katja; de Vos, Wiebe Matthijs; Kamperman, Marleen

    2017-01-01

    The incorporation of stimuli-responsive pores in nanoporous membranes is a promising approach to facilitate the cleaning process of the membranes. Here we present fully reversible thermo-responsive nanoporous membranes fabricated by self-assembly and non-solvent induced phase separation (SNIPS) of

  17. High Molecular Weight Glucan of the Culinary Medicinal Mushroom Agaricus bisporus is an α-Glucan that Forms Complexes with Low Molecular Weight Galactan

    Directory of Open Access Journals (Sweden)

    Harry J. Wichers

    2010-08-01

    Full Text Available An a-glucan was isolated from the culinary medicinal mushroom A. bisporus by hot water extraction, ethanol precipitation and DEAE-cellulose chromatography. The resulting material showed a single HMW peak excluded from a Sephadex G50 column that could completely be degraded by α-amylase treatment. After heating in 1% SDS a small additional peak of low MW eluted from the G50 column. The monosaccharide composition of the main peak was evaluated by HPLC, and was found to consist of a majority of glucose (97.6%, and a minor proportion of galactose (2.4%. Methylation analysis and degradation by a-amylase indicated the presence of an a-glucan with a main chain consisting of (1®4-linked units, substituted at O-6 by α-D-glucopyranose single-units in the relation 1:8. Mono- (13C-, 1H-NMR and bidimensional [1H (obs.,13C-HSQC] spectroscopy analysis confirmed the a-configuration of the Glcp residues by low frequency resonances of C-1 at d 100.6, 100.2, and 98.8 ppm and H-1 high field ones at d 5.06, 5.11, and 4.74 ppm. The DEPT-13C-NMR allowed assigning the non-substituted and O-substituted –CH2 signals at d 60.3/60.8 and 66.2 ppm, respectively. Other assignments were attributed to C-2, C-3, C-4, C-5 and C-6 of the non-reducing ends at d 71.8; 72.8; 70.0; 71.3 and 60.3/60.8 ppm, respectively. The minor proportion of galactose that was demonstrated was probably derived from a complex between the a-glucan and a low molecular weight galactan.

  18. Monolithic poly(1,2-bis(p-vinylphenyl)ethane) capillary columns for simultaneous separation of low- and high-molecular-weight compounds.

    Science.gov (United States)

    Greiderer, Andreas; Ligon, S Clark; Huck, Christian W; Bonn, Günther K

    2009-08-01

    Monolithic poly(1,2-bis(p-vinylphenyl)ethane (BVPE)) capillary columns were prepared by thermally initiated free radical polymerisation of 1,2-bis(p-vinylphenyl)ethane in the presence of inert diluents (porogens) and alpha,alpha'-azoisobutyronitrile (AIBN) as initiator. Polymerisations were accomplished in 200 microm ID fused silica capillaries at 65 degrees C and for 60 min. Mercury intrusion porosimetry measurements of the polymeric RP support showed a broad bimodal pore-size-distribution of mesopores and small macropores in the range of 5-400 nm and flow-channels in the mum range. N(2)-adsorption (BET) analysis resulted in a tremendous enhancement of surface area (101 m(2)/g) of BVPE stationary phases compared to typical organic monoliths (approximately 20 m(2)/g), indicating the presence of a considerable amount of mesopores. Consequently, the adequate proportion of both meso- and (small) macropores allowed the rapid and high-resolution separation of low-molecular-weight compounds as well as biomolecules on the same monolithic support. At the same time, the high fraction of flow-channels provided enhanced column permeability. The chromatographic performance of poly(1,2-bis(p-vinylphenyl)ethane) capillary columns for the separation of biomolecules (proteins, oligonucleotides) and small molecules (alkyl benzenes, phenols, phenons) are demonstrated in this article. Additionally, pressure drop versus flow rate measurements of novel poly(1,2-bis(p-vinylphenyl)ethane) capillary columns confirmed high mechanical robustness, low swelling in organic solvents and high permeability. Due to the simplicity of monolith fabrication, comprehensive studies of the retention and separation behaviour of monolithic BVPE columns resulted in high run-to-run and batch-to-batch reproducibilities. All these attributes prove the excellent applicability of monolithic poly(1,2-bis(p-vinylphenyl)ethane) capillary columns for micro-HPLC towards a huge range of analytes of different chemistries and molecular sizes.

  19. Stereocomplexation of low molecular weight poly(L-lactic acid) and high molecular weight poly(D-lactic acid), radiation crosslinking PLLA/PDLA stereocomplexes and their characterization

    International Nuclear Information System (INIS)

    Quynh, Tran Minh; Mai, Hoang Hoa; Lan, Pham Ngoc

    2013-01-01

    Poly(L-lactic acid)s (PLLAx) were synthesized from L-lactic acid by polycondensation. Different stereocomplexes were also obtained with equimolar mixtures of synthesized PLLAx and a commercial PDLA. The stereocomplexes were crosslinked with triallyl isocyanurate (TAIC) by gamma irradiation. Crosslinking density increased with radiation doses, the heavier the crosslinking network, the lower its swelling degree. The crosslinking structures were introduced in the stereocomplexes inhibiting the mobility for crystallization of PLLA molecules. Thermal and mechanical properties of PLA stereocomplexes were remarkably enhanced by radiation induced crosslinking. PLA stereocomplex does not seem to be degraded by PLLA degrading microorganisms existing in compost at room temperature, but the synthesized PLLA was significantly degraded. - Highlights: ► Complete PLA stereocomplex was obtained from synthesized PLLA and a commercial PDLA. ► Melting temperature of stereocomplex were much improved by gamma irradiation. ► Crosslinking network inhibited the mobility of polymeric chains for crystallization. ► Biodegradability of PLLA was reduced by stereocomplexation and crosslinking.

  20. Bioremediation of high molecular weight polyaromatic hydrocarbons co-contaminated with metals in liquid and soil slurries by metal tolerant PAHs degrading bacterial consortium.

    Science.gov (United States)

    Thavamani, Palanisami; Megharaj, Mallavarapu; Naidu, Ravi

    2012-11-01

    Bioremediation of polyaromatic hydrocarbons (PAH) contaminated soils in the presence of heavy metals have proved to be difficult and often challenging due to the ability of toxic metals to inhibit PAH degradation by bacteria. In this study, a mixed bacterial culture designated as consortium-5 was isolated from a former manufactured gas plant (MGP) site. The ability of this consortium to utilise HMW PAHs such as pyrene and BaP as a sole carbon source in the presence of toxic metal Cd was demonstrated. Furthermore, this consortium has proven to be effective in degradation of HMW PAHs even from the real long term contaminated MGP soil. Thus, the results of this study demonstrate the great potential of this consortium for field scale bioremediation of PAHs in long term mix contaminated soils such as MGP sites. To our knowledge this is the first study to isolate and characterize metal tolerant HMW PAH degrading bacterial consortium which shows great potential in bioremediation of mixed contaminated soils such as MGP.

  1. Enhancement of human adaptive immune responses by administration of a high-molecular-weight polysaccharide extract from the cyanobacterium Arthrospira platensis

    DEFF Research Database (Denmark)

    Løbner, Morten; Walsted, Anette; Larsen, Rune

    2008-01-01

    administration for 3 days (P production persisted after 56 days (P ... production of the Th1 cytokines tumor necrosis factor (TNF)-alpha, interleukin (IL)-2, and interferon (IFN)-gamma was increased after Immunlina...

  2. Formation of high-molecular-weight angiotensinogen during pregnancy is a result of competing redox reactions with the proform of eosinophil major basic protein

    DEFF Research Database (Denmark)

    Kløverpris, Søren; Skov, Louise Lind; Glerup, Simon

    2013-01-01

    compared to monomeric AGT and the proMBP-AGT complex. Furthermore, we have used recombinant proteins to analyse the formation of the proMBP-PAPP-A and the proMBP-AGT complexes, and we demonstrate that they are competing reactions, depending on the same cysteine residue of proMBP, but differentially...... on the redox potential, potentially important for the relative amounts of the complexes in vivo. These findings may be important physiologically, since the biochemical properties of the proteins change as a consequence of complex formation....

  3. M/sub r/ 25,000 heparin-binding protein from guinea pig brain is a high molecular weight form of basic fibroblast growth factor

    International Nuclear Information System (INIS)

    Moscatelli, D.; Joseph-Silverstein, J.; Manejias, R.; Rifkin, D.B.

    1987-01-01

    A M/sub r/ 25,000 form of basic fibroblast growth factor (bFGF) has been isolated from guinea pig grain along with the typical M/sub r/ 18,000 form. Both forms were purified to homogeneity by a combination of heparin-affinity chromatography and ion-exchange chromatography on an FPLC Mono S column. The M/sub r/ 25,000 form, like the M/sub r/ 18,000 form was not eluted from the heparin-affinity column with 0.95 M NaCl, but was eluted with 2 M NaCl. The M/sub r/ 25,000 guinea pig protein stimulated plasminogen activator production by cultured bovine capillary endothelial cells in a dose-dependent manner at concentration of 0.1-10 ngml, the same range that was effective for guinea pig and human M/sub r/ 18,000 bFGFs. The binding of human 125 I-labeled bFGF to baby hamster kidney cells is inhibited equally by the M/sub r/ 25,000 guinea pig protein and the M/sub r/ 18,000 guinea pig and human bFGFs. Polyclonal antibodies raised against human bFGF recognize both the M/sub r/ 25,000 and 18,000 guinea pig proteins in an immunoblot analysis. In a radioimmunoassay, both the M/sub r/ 25,000 and M/sub r/ 18,000 guinea pig proteins compete equally well with iodinated human bFGF for binding to the anti-human bFGF antibodies. When treated with low concentrations of trypsin, the M/sub r/ 25,000 guinea pig bFGF was converted to a M/sub r/ 18,000 protein. These results show that the two molecules are closely related and suggest that the M/sub r/ 25,000 protein shares substantial homology with the M/sub r/ 18,000 bFGF

  4. Mr 25,000 heparin-binding protein from guinea pig brain is a high molecular weight form of basic fibroblast growth factor.

    OpenAIRE

    Moscatelli, D; Joseph-Silverstein, J; Manejias, R; Rifkin, D B

    1987-01-01

    A Mr 25,000 form of basic fibroblast growth factor (bFGF) has been isolated from guinea pig brain along with the typical Mr 18,000 form. Both forms were purified to homogeneity by a combination of heparin-affinity chromatography and ion-exchange chromatography on an FPLC Mono S column. The Mr 25,000 form, like the Mr 18,000 form, was not eluted from the heparin-affinity column with 0.95 M NaCl, but was eluted with 2 M NaCl. The Mr 25,000 guinea pig protein stimulated plasminogen activator pro...

  5. Vitellogenins Are New High Molecular Weight Components and Allergens (Api m 12 and Ves v 6) of Apis mellifera and Vespula vulgaris Venom

    Science.gov (United States)

    Blank, Simon; Seismann, Henning; McIntyre, Mareike; Ollert, Markus; Wolf, Sara; Bantleon, Frank I.; Spillner, Edzard

    2013-01-01

    Background/Objectives Anaphylaxis due to hymenoptera stings is one of the most severe clinical outcomes of IgE-mediated hypersensitivity reactions. Although allergic reactions to hymenoptera stings are often considered as a general model for the underlying principles of allergic disease, venom immunotherapy is still hampered by severe systemic side effects and incomplete protection. The identification and detailed characterization of all allergens of hymenoptera venoms might result in an improvement in this field and promote the detailed understanding of the allergological mechanism. Our aim was the identification and detailed immunochemical and allergological characterization of the low abundant IgE-reactive 200 kDa proteins of Apis mellifera and Vespula vulgaris venom. Methods/Principal Findings Tandem mass spectrometry-based sequencing of a 200 kDa venom protein yielded peptides that could be assigned to honeybee vitellogenin. The coding regions of the honeybee protein as well as of the homologue from yellow jacket venom were cloned from venom gland cDNA. The newly identified 200 kDa proteins share a sequence identity on protein level of 40% and belong to the family of vitellogenins, present in all oviparous animals, and are the first vitellogenins identified as components of venom. Both vitellogenins could be recombinantly produced as soluble proteins in insect cells and assessed for their specific IgE reactivity. The particular vitellogenins were recognized by approximately 40% of sera of venom-allergic patients even in the absence of cross-reactive carbohydrate determinants. Conclusion With the vitellogenins of Apis mellifera and Vespula vulgaris venom a new homologous pair of venom allergens was identified and becomes available for future applications. Due to their allergenic properties the honeybee and the yellow jacket venom vitellogenin were designated as allergens Api m 12 and Ves v 6, respectively. PMID:23626765

  6. CFG-7-P3 : potential of aggregate-associated biodegradation of high-molecular-weight hydrocarbon fractions in crude-oil contaminated soils from a northern Canadian site

    Energy Technology Data Exchange (ETDEWEB)

    Chang, W.; Snelgrove, J.; Akbari, A.; Ghoshal, S. [McGill Univ., Montreal, PQ (Canada). Dept. of Civil Engineering and Applied Mechanics

    2010-07-01

    Soil aggregation can limit aerobic hydrocarbon biodegradation rates due to the slower intra-pore diffusion of nutrients, oxygen and hydrocarbons. This study investigated the influence of soil aggregation at a pilot-scale biopile of crude oil-contaminated soil shipped from a site in the Northwest Territories. Attempts were made to stimulate indigenous microbial activity of the hydrocarbon-degrading bacteria through soil aeration and nutrient amendments in a tank maintained at 15 degrees C. Results showed that nutrient amendment significantly enhanced aggregation. After 60 days, approximately 50 per cent of the initial total hydrocarbon productivity (TPH) was reduced in both the treated and untreated biopile. However, a TPH analysis of soil aggregate levels showed that the biodegradation of high weight hydrocarbon fractions in macroaggregates was more significantly reduced in the nutrient-amended soils. Results suggested that the soil particles in the macroaggregates were more loosely clustered, and may have supported enhanced hydrocarbon biodegradation.

  7. Reduction Levels and the Effects of High-Molecular-Weight Adiponectin via AMPK/eNOS in Chinese Type 2 Diabetes.

    Science.gov (United States)

    Lian, K; Guo, X; Huang, Q; Tan, Z; Xia, C; Wang, R; Yi, F; Li, W; Liu, J; Li, Y; Guo, S; Wang, H; Ji, Q; Tao, L

    2016-10-01

    Aim: This study was to investigate the change of high-molecula-weight (HMW) adiponectin (APN) isoform, the association between type 2 diabetes mellitus (T2DM) and HMW APN isoform, the variation of Disulfide-bond A oxidoreductase-like protein (DsbA-L), the effect of HMW APN isoform on AMP-dependent protein kinase (AMPK) and endothelial nitric oxide synthase (eNOS) in Chinese T2DM. Method: 169 patients aged at (48.7±9.4) years and 107 healthy control subjects aged at (42.6±7.8) years took part in this study. Anthropometric measures of the characters were assayed and different APN isoforms, DsbA-L, AMPK and eNOS levels were determined. Results: Ln(sRAGE) and Ln(Adiponectin) were significantly lower and significantly higher for the other characteristics in T2DM. Ln(Adiponectin) was negatively and significantly correlated with WHR, Ln(triglycerides), fasting plasma glucose, HbA1c (%) in control subjects and T2DM patients. Plasma and adipose tissue total APN and HMW APN were significantly reduced in newly diagnosed T2DM patients. DsbA-L was markedly down-regulated in diabetic adipose tissue. HMW APN caused significant decreases in AMPK and eNOS phosphorylation levels of human umbilical vein endothelial cells (HUVECs). Conclusions: Our results demonstrated that total APN levels was closely related to the risk of T2DM and HMW APN reduction was involved in the diabetic vascular AMPK/eNOS signal pathway. The findings will provide insight into novel therapeutic approaches for reducing the elevated cardiovascular risk associated with T2DM. © Georg Thieme Verlag KG Stuttgart · New York.

  8. Influence of load and sliding velocity on wear resistance of solid-lubricant composites of ultra-high molecular weight polyethylene

    Science.gov (United States)

    Panin, S. V.; Kornienko, L. A.; Buslovich, D. G.; Alexenko, V. O.; Ivanova, L. R.

    2017-12-01

    To determine the limits of the operation loading intervals appropriate for the use of solid lubricant UHMWPE composites in tribounits for mechanical engineering and medicine, the tribotechnical properties of UHMWPE blends with the optimum solid lubricant filler content (polytetrafluoroethylene, calcium stearate, molybdenum disulfide, colloidal graphite, boron nitride) are studied under dry sliding friction at different velocities (V = 0.3 and 0.5 m/s) and loads (P = 60 and 140 N). It is shown that the wear resistance of solid lubricant UHMWPE composites at moderate sliding velocities (V = 0.3 m/s) and loads (P = 60 N) increases 2-3 times in comparison with pure UHMWPE, while at high load P = 140 N wear resistance of both neat UHMWPE and its composites is reduced almost twice. At high sliding velocities and loads (up to P = 140 N), multiple increasing of the wear of pure UHMWPE and its composites takes place (by the factor of 5 to 10). The operational conditions of UHMWPE composites in tribounits in engineering and medicine are discussed.

  9. Electrophysiology and Beyond: Multiple roles of Na+ channel β subunits in development and disease

    Science.gov (United States)

    Patino, Gustavo A.; Isom, Lori L.

    2010-01-01

    Voltage-gated Na+ channel (VGSC) β subunits are not “auxiliary.” These multifunctional molecules not only modulate Na+ current (INa), but also function as cell adhesion molecules (CAMs) – playing roles in aggregation, migration, invasion, neurite outgrowth, and axonal fasciculation. β subunits are integral members of VGSC signaling complexes at nodes of Ranvier, axon initial segments, and cardiac intercalated disks, regulating action potential propagation through critical intermolecular and cell-cell communication events. At least in vitro, many β subunit cell adhesive functions occur both in the presence and absence of pore-forming VGSC α subunits, and in vivo β subunits are expressed in excitable as well as non-excitable cells, thus β subunits may play important functional roles on their own, in the absence of α subunits. VGSC β1 subunits are essential for life and appear to be especially important during brain development. Mutations in β subunit genes result in a variety of human neurological and cardiovascular diseases. Moreover, some cancer cells exhibit alterations in β subunit expression during metastasis. In short, these proteins, originally thought of as merely accessory to α subunits, are critical players in their own right in human health and disease. Here we discuss the role of VGSC β subunits in the nervous system. PMID:20600605

  10. Architecture of the large subunit of the mammalian mitochondrial ribosome.

    Science.gov (United States)

    Greber, Basil J; Boehringer, Daniel; Leitner, Alexander; Bieri, Philipp; Voigts-Hoffmann, Felix; Erzberger, Jan P; Leibundgut, Marc; Aebersold, Ruedi; Ban, Nenad

    2014-01-23

    Mitochondrial ribosomes synthesize a number of highly hydrophobic proteins encoded on the genome of mitochondria, the organelles in eukaryotic cells that are responsible for energy conversion by oxidative phosphorylation. The ribosomes in mammalian mitochondria have undergone massive structural changes throughout their evolution, including ribosomal RNA shortening and acquisition of mitochondria-specific ribosomal proteins. Here we present the three-dimensional structure of the 39S large subunit of the porcine mitochondrial ribosome determined by cryo-electron microscopy at 4.9 Å resolution. The structure, combined with data from chemical crosslinking and mass spectrometry experiments, reveals the unique features of the 39S subunit at near-atomic resolution and provides detailed insight into the architecture of the polypeptide exit site. This region of the mitochondrial ribosome has been considerably remodelled compared to its bacterial counterpart, providing a specialized platform for the synthesis and membrane insertion of the highly hydrophobic protein components of the respiratory chain.

  11. Protein kinase A regulatory subunit distribution in medulloblastoma

    International Nuclear Information System (INIS)

    Mucignat-Caretta, Carla; Denaro, Luca; Redaelli, Marco; D'Avella, Domenico; Caretta, Antonio

    2010-01-01

    Previous studies showed a differential distribution of the four regulatory subunits of cAMP-dependent protein kinases inside the brain, that changed in rodent gliomas: therefore, the distribution of these proteins inside the brain can give information on the functional state of the cells. Our goal was to examine human brain tumors to provide evidence for a differential distribution of protein kinase A in different tumors. The distribution of detergent insoluble regulatory (R1 and R2) and catalytic subunits of cAMP dependent kinases was examined in pediatric brain tumors by immunohistochemistry and fluorescent cAMP analogues binding. R2 is organized in large single dots in medulloblastomas, while it has a different appearance in other tumors. Fluorescent cAMP labelling was observed only in medulloblastoma. A different distribution of cAMP dependent protein kinases has been observed in medulloblastoma

  12. Testing experimental subunit furunculosis vaccines for rainbow trout

    DEFF Research Database (Denmark)

    Marana, Moonika H.; Chettri, Jiwan Kumar; Skov, Jakob

    2016-01-01

    Aeromonas salmonicida subsp. salmonicida (AS) is the etiological agent of typical furunculosis in salmonid fish. The disease causes bacterial septicemia and is a major fish health problem in salmonid aquaculture worldwide, inducing high morbidity and mortality. In this study we vaccinated rainbow...... trout with subunit vaccines containing protein antigens that were selected based on an in silico antigen discovery approach. Thus, the proteome of AS strain A449 was analyzed by an antigen discovery platform and its proteins consequently ranked by their predicted ability to evoke protective immune...... response against AS. Fourteen proteins were prepared in 3 different experimental subunit vaccine combinations and used to vaccinate rainbow trout by intraperitoneal (i.p.) injection. We tested the proteins for their ability to elicit antibody production and protection. Thus, fish were exposed to virulent...

  13. Mapping of the Mouse Actin Capping Protein Beta Subunit Gene

    Directory of Open Access Journals (Sweden)

    Cooper John A

    2000-07-01

    Full Text Available Abstract Background Capping protein (CP, a heterodimer of α and β subunits, is found in all eukaryotes. CP binds to the barbed ends of actin filaments in vitro and controls actin assembly and cell motility in vivo. Vertebrates have three isoforms of CPβ produced by alternatively splicing from one gene; lower organisms have one gene and one isoform. Results We isolated genomic clones corresponding to the β subunit of mouse CP and identified its chromosomal location by interspecies backcross mapping. Conclusions The CPβ gene (Cappb1 mapped to Chromosome 4 between Cdc42 and D4Mit312. Three mouse mutations, snubnose, curly tail, and cribriform degeneration, map in the vicinity of the β gene.

  14. ASIC subunit ratio and differential surface trafficking in the brain.

    Science.gov (United States)

    Wu, Junjun; Xu, Yuanyuan; Jiang, Yu-Qing; Xu, Jiangping; Hu, Youjia; Zha, Xiang-ming

    2016-01-08

    Acid-sensing ion channels (ASICs) are key mediators of acidosis-induced responses in neurons. However, little is known about the relative abundance of different ASIC subunits in the brain. Such data are fundamental for interpreting the relative contribution of ASIC1a homomers and 1a/2 heteromers to acid signaling, and essential for designing therapeutic interventions to target these channels. We used a simple biochemical approach and semi-quantitatively determined the molar ratio of ASIC1a and 2 subunits in mouse brain. Further, we investigated differential surface trafficking of ASIC1a, ASIC2a, and ASIC2b. ASIC1a subunits outnumber the sum of ASIC2a and ASIC2b. There is a region-specific variation in ASIC2a and 2b expression, with cerebellum and striatum expressing predominantly 2b and 2a, respectively. Further, we performed surface biotinylation and found that surface ASIC1a and ASIC2a ratio correlates with their total expression. In contrast, ASIC2b exhibits little surface presence in the brain. This result is consistent with increased co-localization of ASIC2b with an ER marker in 3T3 cells. Our data are the first semi-quantitative determination of relative subunit ratio of various ASICs in the brain. The differential surface trafficking of ASICs suggests that the main functional ASICs in the brain are ASIC1a homomers and 1a/2a heteromers. This finding provides important insights into the relative contribution of various ASIC complexes to acid signaling in neurons.

  15. Global Proteome Analysis Identifies Active Immunoproteasome Subunits in Human Platelets*

    Science.gov (United States)

    Klockenbusch, Cordula; Walsh, Geraldine M.; Brown, Lyda M.; Hoffman, Michael D.; Ignatchenko, Vladimir; Kislinger, Thomas; Kast, Juergen

    2014-01-01

    The discovery of new functions for platelets, particularly in inflammation and immunity, has expanded the role of these anucleate cell fragments beyond their primary hemostatic function. Here, four in-depth human platelet proteomic data sets were generated to explore potential new functions for platelets based on their protein content and this led to the identification of 2559 high confidence proteins. During a more detailed analysis, consistently high expression of the proteasome was discovered, and the composition and function of this complex, whose role in platelets has not been thoroughly investigated, was examined. Data set mining resulted in identification of nearly all members of the 26S proteasome in one or more data sets, except the β5 subunit. However, β5i, a component of the immunoproteasome, was identified. Biochemical analyses confirmed the presence of all catalytically active subunits of the standard 20S proteasome and immunoproteasome in human platelets, including β5, which was predominantly found in its precursor form. It was demonstrated that these components were assembled into the proteasome complex and that standard proteasome as well as immunoproteasome subunits were constitutively active in platelets. These findings suggest potential new roles for platelets in the immune system. For example, the immunoproteasome may be involved in major histocompatibility complex I (MHC I) peptide generation, as the MHC I machinery was also identified in our data sets. PMID:25146974

  16. Global proteome analysis identifies active immunoproteasome subunits in human platelets.

    Science.gov (United States)

    Klockenbusch, Cordula; Walsh, Geraldine M; Brown, Lyda M; Hoffman, Michael D; Ignatchenko, Vladimir; Kislinger, Thomas; Kast, Juergen

    2014-12-01

    The discovery of new functions for platelets, particularly in inflammation and immunity, has expanded the role of these anucleate cell fragments beyond their primary hemostatic function. Here, four in-depth human platelet proteomic data sets were generated to explore potential new functions for platelets based on their protein content and this led to the identification of 2559 high confidence proteins. During a more detailed analysis, consistently high expression of the proteasome was discovered, and the composition and function of this complex, whose role in platelets has not been thoroughly investigated, was examined. Data set mining resulted in identification of nearly all members of the 26S proteasome in one or more data sets, except the β5 subunit. However, β5i, a component of the immunoproteasome, was identified. Biochemical analyses confirmed the presence of all catalytically active subunits of the standard 20S proteasome and immunoproteasome in human platelets, including β5, which was predominantly found in its precursor form. It was demonstrated that these components were assembled into the proteasome complex and that standard proteasome as well as immunoproteasome subunits were constitutively active in platelets. These findings suggest potential new roles for platelets in the immune system. For example, the immunoproteasome may be involved in major histocompatibility complex I (MHC I) peptide generation, as the MHC I machinery was also identified in our data sets. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  17. Radioimmunoassay of TSH subunits in thyroid diseases and endocrine opthalmopahty

    International Nuclear Information System (INIS)

    Eder, W.

    1982-01-01

    Highly sensitive radioimmunoassays of hTSH sub-units were developed. The hormone preparations were labelled with 125-iodine according to a modified chloramine -T method, and purified by chromatography using biogel P6 and P60. Rabbit antisera were used as antibodies. Separation of the antibody-bound and of the free antigens was carried out via the double antibody method. The antiserum required for this purpose was obtained from a goat. The sensitivity of the assay was influenced by changing the protein content of the buffer, the incubation volume, the tracer amounts, the incubation time and the incubation temperature. For hTSH-α, the lowest detectable limit was found to be 50 pg/ml, for hTSH-#betta# 20 pg/ml. Thus, the sub-units could be determined for 98% of the patients under review. The #betta#-TSH radioimmunoassay is largely specific, TSH cross-reacts to a degree of 5%. The computerized evoluation was carried out by means of Spline approximation using the Siemens 4004 computer. Precision and accurateness are in compliance with generally accpted criteria. The serum levels of α and #betta# sub-units showed no discordancy with regard to TSH. In all groups of patients examined, the levels of the hormone-specific #betta#-chain were found to be exclusively dependent upon the actual thyroid activity. (orig.) [de

  18. Variations in yield and gluten proteins in durum wheat varieties under late-season foliar versus soil application of nitrogen fertilizer in a northern Mediterranean environment.

    Science.gov (United States)

    Visioli, Giovanna; Bonas, Urbana; Dal Cortivo, Cristian; Pasini, Gabriella; Marmiroli, Nelson; Mosca, Giuliano; Vamerali, Teofilo

    2018-04-01

    With the increasing demand for high-quality foodstuffs and concern for environmental sustainability, late-season nitrogen (N) foliar fertilization of common wheat is now an important and widespread practice. This study investigated the effects of late-season foliar versus soil N fertilization on yield and protein content of four varieties of durum wheat, Aureo, Ariosto, Biensur and Liberdur, in a three-year field trial in northern Italy. Variations in low-molecular-weight glutenins (LMW-GS), high-molecular-weight glutenins (HMW-GS) and gliadins were assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). It was found that N applied to the canopy did not improve protein rate compared with N application to the soil (general mean 138 mg g -1 ), but moderately increased productivity in the high-yielding varieties Liberdur and Biensur (three-year means 7.23 vs 7.13 and 7.53 vs 7.09 t ha -1 respectively). Technological quality was mainly related to variety choice, Aureo and Ariosto having higher protein rates and glutenin/gliadin ratios. Also found was a strong 'variety × N application method' interaction in the proportions of protein subunits within each class, particularly LMW-GS and gliadins. A promising result was the higher N uptake efficiency, although as apparent balance, combined with higher HMW/LMW-GS ratio in var. Biensur. Late-season foliar N fertilization allows N fertilizer saving, potentially providing environmental benefits in the rainy climate of the northern Mediterranean area, and also leads to variety-dependent up-regulation of essential LMW-GS and gliadins. Variety choice is a key factor in obtaining high technological quality, although it is currently associated with modest grain yield. This study provides evidence of high quality in the specific high-yielding variety Biensur, suggesting its potential as a mono-varietal semolina for pasta production. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  19. Glutenin composition, quality characteristics, and agronomic ...

    African Journals Online (AJOL)

    ABSTRACT Eleven cultivars of durum wheat (Triticum durum L. var. durum Desf.) ... Gluten strength was measured by the sodium dodecyl sulfate (SDS) sedimentation test. ... Significant genotypic differences were observed for grain yield, 1000 kernel weight, protein content, ... (Af Crop Sci J 2002 Vol 10 No 2 pp173-182).

  20. The cytochrome oxidase subunit I and subunit III genes in Oenothera mitochondria are transcribed from identical promoter sequences

    Science.gov (United States)

    Hiesel, Rudolf; Schobel, Werner; Schuster, Wolfgang; Brennicke, Axel

    1987-01-01

    Two loci encoding subunit III of the cytochrome oxidase (COX) in Oenothera mitochondria have been identified from a cDNA library of mitochondrial transcripts. A 657-bp sequence block upstream from the open reading frame is also present in the two copies of the COX subunit I gene and is presumably involved in homologous sequence rearrangement. The proximal points of sequence rearrangements are located 3 bp upstream from the COX I and 1139 bp upstream from the COX III initiation codons. The 5'-termini of both COX I and COX III mRNAs have been mapped in this common sequence confining the promoter region for the Oenothera mitochondrial COX I and COX III genes to the homologous sequence block. ImagesFig. 5. PMID:15981332

  1. Characterization of wheat varieties by seed storageprotein ...

    African Journals Online (AJOL)

    Journal Home > Vol 6, No 5 (2007) > ... Wheat grains of thirteen varieties were collected from different ecological regions of Pakistan. ... the dendrogram for high molecular weight (HMW) and low molecular weight (LMW) gluten subunit bands.

  2. The trafficking pathway of a wheat storage protein in transgenic rice endosperm.

    Science.gov (United States)

    Oszvald, Maria; Tamas, Laszlo; Shewry, Peter R; Tosi, Paola

    2014-04-01

    The trafficking of proteins in the endoplasmic reticulum (ER) of plant cells is a topic of considerable interest since this organelle serves as an entry point for proteins destined for other organelles, as well as for the ER itself. In the current work, transgenic rice was used to study the pattern and pathway of deposition of the wheat high molecular weight (HMW) glutenin sub-unit (GS) 1Dx5 within the rice endosperm using specific antibodies to determine whether it is deposited in the same or different protein bodies from the rice storage proteins, and whether it is located in the same or separate phases within these. The protein distribution and the expression pattern of HMW sub-unit 1Dx5 in transgenic rice endosperm at different stages of development were determined using light and electron microscopy after labelling with antibodies. The use of HMW-GS-specific antibodies showed that sub-unit 1Dx5 was expressed mainly in the sub-aleurone cells of the endosperm and that it was deposited in both types of protein body present in the rice endosperm: derived from the ER and containing prolamins, and derived from the vacuole and containing glutelins. In addition, new types of protein bodies were also formed within the endosperm cells. The results suggest that the HMW 1Dx5 protein could be trafficked by either the ER or vacuolar pathway, possibly depending on the stage of development, and that its accumulation in the rice endosperm could compromise the structural integrity of protein bodies and their segregation into two distinct populations in the mature endosperm.

  3. Molecular, Physicochemical and Rheological Characteristics of Introgressive Triticale/Triticum monococcum ssp. monococcum Lines with Wheat 1D/1A Chromosome Substitution

    Directory of Open Access Journals (Sweden)

    Lidia Błaszczyk

    2013-07-01

    Full Text Available Three sets of hexaploid introgressive triticale lines, with Triticum monococcum ssp. monococcum (cultivated einkorn wheat genes and a bread wheat chromosome 1D substituted for chromosome 1A, and one set of secondary triticale lines were evaluated for grain and flour physicochemical and dough rheological characteristics in two generations (F7 and F8. Genomic in situ hybridization (GISH and fluorescence in situ hybridization (FISH confirmed the 1D/1A chromosome substitution. The presence or absence of einkorn high-molecular-weight (HMW glutenin subunits and the wheat Glu-D1d locus encoding the 5 + 10 subunits was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE, capillary zone electrophoresis, and allele-specific molecular markers. Significant differences were found among physicochemical properties (with the exception of the Hagberg falling number of all introgressive Triticale/T. monococcum lines and the secondary triticale lines. The wheat 1D/1A chromosome substitution also affected these properties. The results showed that in all introgressive triticale lines, the protein and gluten content, Zeleny sedimentation value, and water absorption capacity, were increased. The rheological parameters estimated using micro-farinograph, reomixer, and Kieffer dough extensibility systems also showed an appreciable increase in dough-mixing properties, maximum resistance to extension (Rmax, and dough extensibility. Introgressive Triticale/T. monococcum lines with 5 + 10 subunits have particularly favorable rheological parameters. The results obtained in this study suggest that the cultivated einkorn genome Am, in the context of hexaploid secondary triticale lines and with a wheat 1D/1A substitution, has the potential to improve gluten polymer interactions and be a valuable genetic resource for triticale quality improvement.

  4. Proteome approaches to characterize seed storage proteins related to ditelocentric chromosomes in common wheat (Triticum aestivum L.).

    Science.gov (United States)

    Islam, Nazrul; Woo, Sun-Hee; Tsujimoto, Hisashi; Kawasaki, Hiroshi; Hirano, Hisashi

    2002-09-01

    Changes in protein composition of wheat endosperm proteome were investigated in 39 ditelocentric chromosome lines of common wheat (Triticum aestivum L.) cv. Chinese Spring. Two-dimensional gel electrophoresis followed by Coomassie Brilliant Blue staining has resolved a total of 105 protein spots in a gel. Quantitative image analysis of protein spots was performed by PDQuest. Variations in protein spots between the euploid and the 39 ditelocentric lines were evaluated by spot number, appearance, disappearance and intensity. A specific spot present in all gels was taken as an internal standard, and the intensity of all other spots was calculated as the ratio of the internal standard. Out of the 1755 major spots detected in 39 ditelocentric lines, 1372 (78%) spots were found variable in different spot parameters: 147 (11%) disappeared, 978 (71%) up-regulated and 247 (18%) down-regulated. Correlation studies in changes in protein intensities among 24 protein spots across the ditelocentric lines were performed. High correlations in changes of protein intensities were observed among the proteins encoded by genes located in the homoeologous arms. Locations of structural genes controlling 26 spots were identified in 10 chromosomal arms. Multiple regulators of the same protein located at various chromosomal arms were also noticed. Identification of structural genes for most of the proteins was found difficult due to multiple regulators encoding the same protein. Two novel subunits (1B(Z,) 1BDz), the structure of which are very similar to the high molecular weight glutenin subunit 12, were identified, and the chromosome arm locations of these subunits were assigned.

  5. Molecular cloning of the human casein kinase II α subunit

    International Nuclear Information System (INIS)

    Meisner, H.; Heller-Harrison, R.; Buxton, J.; Czech, M.P.

    1989-01-01

    A human cDNA encoding the α subunit of casein kinase II and a partial cDNA encoding the rat homologue were isolated by using a Drosophila casein kinase II cDNA probe. The 2.2-kb human cDNA contains a 1.2-kb open reading frame, 150 nucleotides of 5' leader, and 850 nucleotides of 3' noncoding region. Except for the first 7 deduced amino acids that are missing in the rat cDNA, the 328 amino acids beginning with the amino terminus are identical between human and rat. The Drosophila enzyme sequence is 90% identical with the human casein kinase II sequence, and there is only a single amino acid difference between the published partial bovine sequence and the human sequence. In addition, the C-terminus of the human cDNA has an extra 53 amino acids not present in Drosophila. Northern analysis of rat and human RNA showed predominant bands of 5.5, 3.1, and 1.8 kb. In rat tissues, brain and spleen had the highest levels of casein kinase II α subunit specific RNA, while skeletal muscle showed the lowest. Southern analysis of human cultured cell and tissue genomic DNA using the full-length cDNA probe revealed two bands with restriction enzymes that have no recognition sites within the cDNA and three to six bands with enzymes having single internal sites. These results are consistent with the possibility that two genes encode the α subunits

  6. Covalent dimerization of ribulose bisphosphate carboxylase subunits by UV radiation

    Energy Technology Data Exchange (ETDEWEB)

    Ferreira, R.M.B. [Universidade Tecnica, Lisbon (Portugal). Inst. Superior de Agronomia]|[Universidade Nova de Lisboa, Oeiras (Portugal). Instituto de Tecnologia Quimica e Biologica; Franco, E.; Teixeira, A.R.N. [Universidade Tecnica, Lisbon (Portugal). Inst. Superior de Agronomia

    1996-08-15

    The effect of UV radiation (UV-A, UV-B and UV-C) on ribulose bisphosphate carboxylase from a variety of plant species was examined. The exposition of plant leaves or the pure enzyme to UV radiation produced a UV-dependent accumulation of a 65 kDa polypeptide (P65). Different approaches were utilized to elucidate the origin and structure of P65: electrophoretic and fluorographic analyses of {sup 35}S-labelled ribulose biphosphate carboxylase exposed to UV radiation and immunological experiments using antibodies specific for P65, for the large and small subunits of ribulose biphosphate carboxylase and for high-molecular-mass aggregates of the enzyme. These studies revealed that P65 is a dimer, formed by the covalent, non-disulphide linkage of one small subunit with one large subunit of ribulose biphosphate carboxylase. For short periods of time (<1 h), the amount of P65 formed increased with the duration of the exposure to the UV radiation and with the energy of the radiation applied. Prolonged exposure to UV radiation (1-6 h) resulted in the formation of high-molecular-mass aggregates of ribulose biphosphate carboxylase. Formation of P65 was shown to depend on the native state of the protein, was stimulated by inhibitors of enzyme activity, and was inhibited by activators of enzyme activity. A UV-independent accumulation of P65 was also achieved by the in vitro incubation of plant crude extracts. However, the UV-dependent and the UV-independent formation of P65 seemed to occur by distinct molecular mechanisms. The UV-dependent accumulation of P65 was immunologically detected in all species examined, including Lemna minor, Arum italicum, Brassica oleracea, Triticum aestivum, Zea mays, Pisum sativum and Phaseolus vulgaris, suggesting that it may constitute a universal response to UV radiation, common to all photosynthetic tissues. (Author).

  7. Covalent dimerization of ribulose bisphosphate carboxylase subunits by UV radiation

    International Nuclear Information System (INIS)

    Ferreira, R.M.B.; Universidade Nova de Lisboa, Oeiras; Franco, E.; Teixeira, A.R.N.

    1996-01-01

    The effect of UV radiation (UV-A, UV-B and UV-C) on ribulose bisphosphate carboxylase from a variety of plant species was examined. The exposition of plant leaves or the pure enzyme to UV radiation produced a UV-dependent accumulation of a 65 kDa polypeptide (P65). Different approaches were utilized to elucidate the origin and structure of P65: electrophoretic and fluorographic analyses of 35 S-labelled ribulose biphosphate carboxylase exposed to UV radiation and immunological experiments using antibodies specific for P65, for the large and small subunits of ribulose biphosphate carboxylase and for high-molecular-mass aggregates of the enzyme. These studies revealed that P65 is a dimer, formed by the covalent, non-disulphide linkage of one small subunit with one large subunit of ribulose biphosphate carboxylase. For short periods of time (<1 h), the amount of P65 formed increased with the duration of the exposure to the UV radiation and with the energy of the radiation applied. Prolonged exposure to UV radiation (1-6 h) resulted in the formation of high-molecular-mass aggregates of ribulose biphosphate carboxylase. Formation of P65 was shown to depend on the native state of the protein, was stimulated by inhibitors of enzyme activity, and was inhibited by activators of enzyme activity. A UV-independent accumulation of P65 was also achieved by the in vitro incubation of plant crude extracts. However, the UV-dependent and the UV-independent formation of P65 seemed to occur by distinct molecular mechanisms. The UV-dependent accumulation of P65 was immunologically detected in all species examined, including Lemna minor, Arum italicum, Brassica oleracea, Triticum aestivum, Zea mays, Pisum sativum and Phaseolus vulgaris, suggesting that it may constitute a universal response to UV radiation, common to all photosynthetic tissues. (Author)

  8. Flexible Connectors between Capsomer Subunits that Regulate Capsid Assembly.

    Science.gov (United States)

    Hasek, Mary L; Maurer, Joshua B; Hendrix, Roger W; Duda, Robert L

    2017-08-04

    Viruses build icosahedral capsids of specific size and shape by regulating the spatial arrangement of the hexameric and pentameric protein capsomers in the growing shell during assembly. In the T=7 capsids of Escherichia coli bacteriophage HK97 and other phages, 60 capsomers are hexons, while the rest are pentons that are correctly positioned during assembly. Assembly of the HK97 capsid to the correct size and shape has been shown to depend on specific ionic contacts between capsomers. We now describe additional ionic interactions within capsomers that also regulate assembly. Each is between the long hairpin, the "E-loop," that extends from one subunit to the adjacent subunit within the same capsomer. Glutamate E153 on the E-loop and arginine R210 on the adjacent subunit's backbone alpha-helix form salt bridges in hexamers and pentamers. Mutations that disrupt these salt bridges were lethal for virus production, because the mutant proteins assembled into tubes or sheets instead of capsids. X-ray structures show that the E153-R210 links are flexible and maintained during maturation despite radical changes in capsomer shape. The E153-R210 links appear to form early in assembly to enable capsomers to make programmed changes in their shape during assembly. The links also prevent flattening of capsomers and premature maturation. Mutant phenotypes and modeling support an assembly model in which flexible E153-R210 links mediate capsomer shape changes that control where pentons are placed to create normal-sized capsids. The E-loop may be conserved in other systems in order to play similar roles in regulating assembly. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Sex Hormone Receptor Expression in the Human Vocal Fold Subunits.

    Science.gov (United States)

    Kirgezen, Tolga; Sunter, Ahmet Volkan; Yigit, Ozgur; Huq, Gulben Erdem

    2017-07-01

    The study aimed to evaluate the existence of sex hormone receptors in the subunits of vocal fold. This is a cadaver study. The androgen, estrogen, and progesterone receptors were examined in the epithelium (EP), superficial layer of the lamina propria (SLP), vocal ligament (VL), and macula flava (MF) of the vocal folds from 42 human cadavers (21 male, 21 female) by immunohistochemical methods. Their staining ratios were scored and statistically compared. The androgen receptor score was significantly higher for the MF than for the EP and SLP (P vocal fold, mostly in the MF and VLs. Copyright © 2017 The Voice Foundation. Published by Elsevier Inc. All rights reserved.

  10. N-linked glycans are required on epithelial Na+ channel subunits for maturation and surface expression.

    Science.gov (United States)

    Kashlan, Ossama B; Kinlough, Carol L; Myerburg, Michael M; Shi, Shujie; Chen, Jingxin; Blobner, Brandon M; Buck, Teresa M; Brodsky, Jeffrey L; Hughey, Rebecca P; Kleyman, Thomas R

    2018-03-01

    Epithelial Na + channel (ENaC) subunits undergo N-linked glycosylation in the endoplasmic reticulum where they assemble into an αβγ complex. Six, 13, and 5 consensus sites (Asn-X-Ser/Thr) for N-glycosylation reside in the extracellular domains of the mouse α-, β-, and γ-subunits, respectively. Because the importance of ENaC N-linked glycans has not been fully addressed, we examined the effect of preventing N-glycosylation of specific subunits on channel function, expression, maturation, and folding. Heterologous expression in Xenopus oocytes or Fischer rat thyroid cells with αβγ-ENaC lacking N-linked glycans on a single subunit reduced ENaC activity as well as the inhibitory response to extracellular Na + . The lack of N-linked glycans on the β-subunit also precluded channel activation by trypsin. However, channel activation by shear stress was N-linked glycan independent, regardless of which subunit was modified. We also discovered that the lack of N-linked glycans on any one subunit reduced the total and surface levels of cognate subunits. The lack of N-linked glycans on the β-subunit had the largest effect on total levels, with the lack of N-linked glycans on the γ- and α-subunits having intermediate and modest effects, respectively. Finally, channels with wild-type β-subunits were more sensitive to limited trypsin proteolysis than channels lacking N-linked glycans on the β-subunit. Our results indicate that N-linked glycans on each subunit are required for proper folding, maturation, surface expression, and function of the channel.

  11. A charged residue at the subunit interface of PCNA promotes trimer formation by destabilizing alternate subunit interactions

    International Nuclear Information System (INIS)

    Freudenthal, Bret D.; Gakhar, Lokesh; Ramaswamy, S.; Washington, M. Todd

    2009-01-01

    Eukaryotic proliferating cell nuclear antigen (PCNA), an essential accessory factor in DNA replication and repair, is a ring-shaped homotrimer. A novel nontrimeric structure of E113G-mutant PCNA protein is reported, which shows that this protein forms alternate subunit interactions. It is concluded that the charged side chain of Glu113 promotes normal trimer formation by destabilizing these alternate subunit interactions. Eukaryotic proliferating cell nuclear antigen (PCNA) is an essential replication accessory factor that interacts with a variety of proteins involved in DNA replication and repair. Each monomer of PCNA has an N-terminal domain A and a C-terminal domain B. In the structure of the wild-type PCNA protein, domain A of one monomer interacts with domain B of a neighboring monomer to form a ring-shaped trimer. Glu113 is a conserved residue at the subunit interface in domain A. Two distinct X-ray crystal structures have been determined of a mutant form of PCNA with a substitution at this position (E113G) that has previously been studied because of its effect on translesion synthesis. The first structure was the expected ring-shaped trimer. The second structure was an unanticipated nontrimeric form of the protein. In this nontrimeric form, domain A of one PCNA monomer interacts with domain A of a neighboring monomer, while domain B of this monomer interacts with domain B of a different neighboring monomer. The B–B interface is stabilized by an antiparallel β-sheet and appears to be structurally similar to the A–B interface observed in the trimeric form of PCNA. The A–A interface, in contrast, is primarily stabilized by hydrophobic interactions. Because the E113G substitution is located on this hydrophobic surface, the A–A interface should be less favorable in the case of the wild-type protein. This suggests that the side chain of Glu113 promotes trimer formation by destabilizing these possible alternate subunit interactions

  12. Determinants of RNA polymerase alpha subunit for interaction with beta, beta', and sigma subunits: hydroxyl-radical protein footprinting.

    OpenAIRE

    Heyduk, T; Heyduk, E; Severinov, K; Tang, H; Ebright, R H

    1996-01-01

    Escherichia coli RNA polymerase (RNAP) alpha subunit serves as the initiator for RNAP assembly, which proceeds according to the pathway 2 alpha-->alpha 2-->alpha 2 beta-->alpha 2 beta beta'-->alpha 2 beta beta' sigma. In this work, we have used hydroxyl-radical protein footprinting to define determinants of alpha for interaction with beta, beta', and sigma. Our results indicate that amino acids 30-75 of alpha are protected from hydroxyl-radical-mediated proteolysis upon interaction with beta ...

  13. Heterotrimeric G protein subunits are located on rat liver endosomes

    Directory of Open Access Journals (Sweden)

    Van Dyke Rebecca W

    2004-01-01

    Full Text Available Abstract Background Rat liver endosomes contain activated insulin receptors and downstream signal transduction molecules. We undertook these studies to determine whether endosomes also contain heterotrimeric G proteins that may be involved in signal transduction from G protein-coupled receptors. Results By Western blotting Gsα, Giα1,2, Giα3 and Gβ were enriched in both canalicular (CM and basolateral (BLM membranes but also readily detectable on three types of purified rat liver endosomes in the order recycling receptor compartment (RRC > compartment for uncoupling of receptor and ligand (CURL > multivesicular bodies (MVB >> purified secondary lysosomes. Western blotting with antibodies to Na, K-ATPase and to other proteins associated with plasma membranes and intracellular organelles indicated this was not due to contamination of endosome preparations by CM or BLM. Adenylate cyclase (AC was also identified on purified CM, BLM, RRC, CURL and MVB. Percoll gradient fractionation of liver postnuclear supernatants demonstrated co-occurrence of endosomes and heterotrimeric G protein subunits in fractions with little plasma membrane markers. By confocal microscopy, punctate staining for Gsα, Giα3 and Gβ corresponded to punctate areas of endocytosed Texas red-dextran in hepatocytes from control and cholera toxin-treated livers. Conclusion We conclude that heterotrimeric G protein subunits as well as AC likely traffic into hepatocytes on endosome membranes, possibly generating downstream signals spatially separate from signalling generated at the plasma membrane, analogous to the role(s of internalized insulin receptors.

  14. Fungal mediator tail subunits contain classical transcriptional activation domains.

    Science.gov (United States)

    Liu, Zhongle; Myers, Lawrence C

    2015-04-01

    Classical activation domains within DNA-bound eukaryotic transcription factors make weak interactions with coactivator complexes, such as Mediator, to stimulate transcription. How these interactions stimulate transcription, however, is unknown. The activation of reporter genes by artificial fusion of Mediator subunits to DNA binding domains that bind to their promoters has been cited as evidence that the primary role of activators is simply to recruit Mediator. We have identified potent classical transcriptional activation domains in the C termini of several tail module subunits of Saccharomyces cerevisiae, Candida albicans, and Candida dubliniensis Mediator, while their N-terminal domains are necessary and sufficient for their incorporation into Mediator but do not possess the ability to activate transcription when fused to a DNA binding domain. This suggests that Mediator fusion proteins actually are functioning in a manner similar to that of a classical DNA-bound activator rather than just recruiting Mediator. Our finding that deletion of the activation domains of S. cerevisiae Med2 and Med3, as well as C. dubliniensis Tlo1 (a Med2 ortholog), impairs the induction of certain genes shows these domains function at native promoters. Activation domains within coactivators are likely an important feature of these complexes and one that may have been uniquely leveraged by a common fungal pathogen. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  15. Thermostable cross-protective subunit vaccine against Brucella species.

    Science.gov (United States)

    Cherwonogrodzky, John W; Barabé, Nicole D; Grigat, Michelle L; Lee, William E; Poirier, Robert T; Jager, Scott J; Berger, Bradley J

    2014-12-01

    A subunit vaccine candidate was produced from Brucella suis 145 (biovar 4; expressing both the A antigen of Brucella abortus and the M antigen of Brucella melitensis). The preparation consisted mostly of polysaccharide (PS; >90% [wt/wt]; both cell-associated PS and exo-PS were combined) and a small amount of protein (1 to 3%) with no apparent nucleic acids. Vaccinated mice were protected (these had a statistically significant reduction in bacterial colonization compared to that of unvaccinated controls) when challenged with representative strains of three Brucella species most pathogenic for humans, i.e., B. abortus, B. melitensis, and B. suis. As little as 1 ng of the vaccine, without added adjuvant, protected mice against B. suis 145 infection (5 × 10(5) CFU), and a single injection of 1 μg of this subunit vaccine protected mice from B. suis 145 challenge for at least 14 months. A single immunization induced a serum IgG response to Brucella antigens that remained elevated for up to 9 weeks. The use of heat (i.e., boiling-water bath, autoclaving) in the vaccine preparation showed that it was thermostable. This method also ensured safety and security. The vaccine produced was immunogenic and highly protective against multiple strains of Brucella and represents a promising candidate for further evaluation. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  16. Binding of ATP by pertussis toxin and isolated toxin subunits

    International Nuclear Information System (INIS)

    Hausman, S.Z.; Manclark, C.R.; Burns, D.L.

    1990-01-01

    The binding of ATP to pertussis toxin and its components, the A subunit and B oligomer, was investigated. Whereas, radiolabeled ATP bound to the B oligomer and pertussis toxin, no binding to the A subunit was observed. The binding of [ 3 H]ATP to pertussis toxin and the B oligomer was inhibited by nucleotides. The relative effectiveness of the nucleotides was shown to be ATP > GTP > CTP > TTP for pertussis toxin and ATP > GTP > TTP > CTP for the B oligomer. Phosphate ions inhibited the binding of [ 3 H]ATP to pertussis toxin in a competitive manner; however, the presence of phosphate ions was essential for binding of ATP to the B oligomer. The toxin substrate, NAD, did not affect the binding of [ 3 H]ATP to pertussis toxin, although the glycoprotein fetuin significantly decreased binding. These results suggest that the binding site for ATP is located on the B oligomer and is distinct from the enzymatically active site but may be located near the eukaryotic receptor binding site

  17. Binding of ATP by pertussis toxin and isolated toxin subunits

    Energy Technology Data Exchange (ETDEWEB)

    Hausman, S.Z.; Manclark, C.R.; Burns, D.L. (Center for Biologics Evaluation and Research, Bethesda, MD (USA))

    1990-07-03

    The binding of ATP to pertussis toxin and its components, the A subunit and B oligomer, was investigated. Whereas, radiolabeled ATP bound to the B oligomer and pertussis toxin, no binding to the A subunit was observed. The binding of ({sup 3}H)ATP to pertussis toxin and the B oligomer was inhibited by nucleotides. The relative effectiveness of the nucleotides was shown to be ATP > GTP > CTP > TTP for pertussis toxin and ATP > GTP > TTP > CTP for the B oligomer. Phosphate ions inhibited the binding of ({sup 3}H)ATP to pertussis toxin in a competitive manner; however, the presence of phosphate ions was essential for binding of ATP to the B oligomer. The toxin substrate, NAD, did not affect the binding of ({sup 3}H)ATP to pertussis toxin, although the glycoprotein fetuin significantly decreased binding. These results suggest that the binding site for ATP is located on the B oligomer and is distinct from the enzymatically active site but may be located near the eukaryotic receptor binding site.

  18. Role of the beta subunit of casein kinase-2 on the stability and specificity of the recombinant reconstituted holoenzyme

    DEFF Research Database (Denmark)

    Meggio, F; Boldyreff, B; Marin, O

    1992-01-01

    Recombinant human alpha subunit from casein kinase-2 (CK-2) was subjected, either alone or in combination with recombinant human beta subunit, to high temperature, tryptic digestion and urea treatment. In all three cases, it was shown that the presence of the beta subunit could drastically reduce...... the autophosphorylation site. It is suggested that the acidic domain of the beta subunit, encompassing residues 55-71, plays a role in the interactions between the beta and alpha subunits....

  19. Efficient expression of functional (α6β22β3 AChRs in Xenopus oocytes from free subunits using slightly modified α6 subunits.

    Directory of Open Access Journals (Sweden)

    Carson Kai-Kwong Ley

    Full Text Available Human (α6β2(α4β2β3 nicotinic acetylcholine receptors (AChRs are essential for addiction to nicotine and a target for drug development for smoking cessation. Expressing this complex AChR is difficult, but has been achieved using subunit concatamers. In order to determine what limits expression of α6* AChRs and to efficiently express α6* AChRs using free subunits, we investigated expression of the simpler (α6β22β3 AChR. The concatameric form of this AChR assembles well, but is transported to the cell surface inefficiently. Various chimeras of α6 with the closely related α3 subunit increased expression efficiency with free subunits and produced pharmacologically equivalent functional AChRs. A chimera in which the large cytoplasmic domain of α6 was replaced with that of α3 increased assembly with β2 subunits and transport of AChRs to the oocyte surface. Another chimera replacing the unique methionine 211 of α6 with leucine found at this position in transmembrane domain 1 of α3 and other α subunits increased assembly of mature subunits containing β3 subunits within oocytes. Combining both α3 sequences in an α6 chimera increased expression of functional (α6β22β3 AChRs to 12-fold more than with concatamers. This is pragmatically useful, and provides insights on features of α6 subunit structure that limit its expression in transfected cells.

  20. Characterisation by nuclear magnetic resonance of the β catalytic subunit of the chloroplastic coupling factor

    International Nuclear Information System (INIS)

    Andre, Francois

    1986-09-01

    This academic work addressed the use of nuclear magnetic resonance (NMR) for the structural and dynamic study of the catalytic sub-unit of the extrinsic section of a membrane complex, the chloroplastic H+-ATPase. This work included the development of a protocol of preparation and quantitative purification of β subunits isolated from the CF1 for the elaboration of a concentrated sample for NMR, and then the study of the β subunit by using proton NMR

  1. Antibodies to the α-subunit of insulin receptor from eggs of immunized hens

    International Nuclear Information System (INIS)

    Song, C.; Yu, J.; Bai, D.H.; Hester, P.Y.; Kim, K.

    1985-01-01

    Simple methods for the generation, purification, and assay of antibodies to the α-subunit of insulin receptor from eggs of immunized hen have been described. Chicken antibodies against the α-subunit inhibit insulin binding to the receptor and stimulate glucose oxidation as well as autophosphorylation of the β-subunit. Thus the properties of chicken antibodies are very similar to those of antibodies found in human autoimmune diseases and different from rabbit antibodies obtained against the same antigen

  2. Cloning and sequencing of the casein kinase 2 alpha subunit from Zea mays

    DEFF Research Database (Denmark)

    Dobrowolska, G; Boldyreff, B; Issinger, O G

    1991-01-01

    The nucleotide sequence of the cDNA coding for the alpha subunit of casein kinase 2 of Zea mays has been determined. The cDNA clone contains an open reading frame of 996 nucleotides encoding a polypeptide comprising 332 amino acids. The primary amino acid sequence exhibits 75% identity to the alpha...... subunit and 71% identity to the alpha' subunit of human casein kinase 2....

  3. Compensatory expression of human -Acetylglucosaminyl-1-phosphotransferase subunits in mucolipidosis type III gamma

    OpenAIRE

    Pohl , Sandra; Tiede , Stephan; Castrichini , Monica; Cantz , Michael; Gieselmann , Volkmar; Braulke , Thomas

    2009-01-01

    Abstract The N-Acetylglucosaminyl-1-phosphotransferase plays a key role in the generation of mannose 6-phosphate (M6P) recognition markers essential for efficient transport of lysosomal hydrolases to lysosomes. The phosphotransferase is composed of six subunits (?2, ?2, ?2). The ?- and ?-subunits are catalytically active and encoded by a single gene, GNPTAB, whereas the ?-subunit encoded by GNPTG is proposed to recognize conformational structures common to lysosomal enzymes. Defects in GN...

  4. Subunits of the Snf1 kinase heterotrimer show interdependence for association and activity.

    Science.gov (United States)

    Elbing, Karin; Rubenstein, Eric M; McCartney, Rhonda R; Schmidt, Martin C

    2006-09-08

    The Snf1 kinase and its mammalian orthologue, the AMP-activated protein kinase (AMPK), function as heterotrimers composed of a catalytic alpha-subunit and two non-catalytic subunits, beta and gamma. The beta-subunit is thought to hold the complex together and control subcellular localization whereas the gamma-subunit plays a regulatory role by binding to and blocking the function of an auto-inhibitory domain (AID) present in the alpha-subunit. In addition, catalytic activity requires phosphorylation by a distinct upstream kinase. In yeast, any one of three Snf1-activating kinases, Sak1, Tos3, or Elm1, can fulfill this role. We have previously shown that Sak1 is the only Snf1-activating kinase that forms a stable complex with Snf1. Here we show that the formation of the Sak1.Snf1 complex requires the beta- and gamma-subunits in vivo. However, formation of the Sak1.Snf1 complex is not necessary for glucose-regulated phosphorylation of the Snf1 activation loop. Snf1 kinase purified from cells lacking the beta-subunits do not contain any gamma-subunit, indicating that the Snf1 kinase does not form a stable alphagamma dimer in vivo. In vitro kinase assays using purified full-length and truncated Snf1 proteins demonstrate that the kinase domain, which lacks the AID, is significantly more active than the full-length Snf1 protein. Addition of purified beta- and gamma-subunits could stimulate the kinase activity of the full-length alpha-subunit but only when all three subunits were present, suggesting an interdependence of all three subunits for assembly of a functional complex.

  5. The testis-specific Cα2 subunit of PKA is kinetically indistinguishable from the common Cα1 subunit of PKA

    Directory of Open Access Journals (Sweden)

    Herberg Friedrich W

    2011-08-01

    Full Text Available Abstract Background The two variants of the α-form of the catalytic (C subunit of protein kinase A (PKA, designated Cα1 and Cα2, are encoded by the PRKACA gene. Whereas Cα1 is ubiquitous, Cα2 expression is restricted to the sperm cell. Cα1 and Cα2 are encoded with different N-terminal domains. In Cα1 but not Cα2 the N-terminal end introduces three sites for posttranslational modifications which include myristylation at Gly1, Asp-specific deamidation at Asn2 and autophosphorylation at Ser10. Previous reports have implicated specific biological features correlating with these modifications on Cα1. Since Cα2 is not modified in the same way as Cα1 we tested if they have distinct biochemical activities that may be reflected in different biological properties. Results We show that Cα2 interacts with the two major forms of the regulatory subunit (R of PKA, RI and RII, to form cAMP-sensitive PKAI and PKAII holoenzymes both in vitro and in vivo as is also the case with Cα1. Moreover, using Surface Plasmon Resonance (SPR, we show that the interaction patterns of the physiological inhibitors RI, RII and PKI were comparable for Cα2 and Cα1. This is also the case for their potency to inhibit catalytic activities of Cα2 and Cα1. Conclusion We conclude that the regulatory complexes formed with either Cα1 or Cα2, respectively, are indistinguishable.

  6. The NH2-terminal php domain of the alpha subunit of the Escherichia coli replicase binds the epsilon proofreading subunit.

    Science.gov (United States)

    Wieczorek, Anna; McHenry, Charles S

    2006-05-05

    The alpha subunit of the replicase of all bacteria contains a php domain, initially identified by its similarity to histidinol phosphatase but of otherwise unknown function (Aravind, L., and Koonin, E. V. (1998) Nucleic Acids Res. 26, 3746-3752). Deletion of 60 residues from the NH2 terminus of the alpha php domain destroys epsilon binding. The minimal 255-residue php domain, estimated by sequence alignment with homolog YcdX, is insufficient for epsilon binding. However, a 320-residue segment including sequences that immediately precede the polymerase domain binds epsilon with the same affinity as the 1160-residue full-length alpha subunit. A subset of mutations of a conserved acidic residue (Asp43 in Escherichia coli alpha) present in the php domain of all bacterial replicases resulted in defects in epsilon binding. Using sequence alignments, we show that the prototypical gram+ Pol C, which contains the polymerase and proofreading activities within the same polypeptide chain, has an epsilon-like sequence inserted in a surface loop near the center of the homologous YcdX protein. These findings suggest that the php domain serves as a platform to enable coordination of proofreading and polymerase activities during chromosomal replication.

  7. Distinct forms of the β subunit of GTP-binding regulatory proteins identified by molecular cloning

    International Nuclear Information System (INIS)

    Fong, H.K.W.; Amatruda, T.T. III; Birren, B.W.; Simon, M.I.

    1987-01-01

    Two distinct β subunits of guanine nucleotide-binding regulatory proteins have been identified by cDNA cloning and are referred to as β 1 and β 1 subunits. The bovine transducin β subunit (β 1 ) has been cloned previously. The author now isolated and analyzed cDNA clones that encode the β 2 subunit from bovine adrenal, bovine brain, and a human myeloid leukemia cell line, HL-60. The 340-residue M/sub r/ 37,329 Β 2 protein is 90% identical with β 1 in predicted amino acid sequence, and it is also organized as a series of repetitive homologous segments. The major mRNA that encodes the bovine β 2 subunit is 1.7 kilobases in length. It is expressed at lower levels than β 1 subunit mRNA in all tissues examined. The β 1 and β 2 messages are expressed in cloned human cell lines. Hybridization of cDNA probes to bovine DNA showed that β 1 and β 2 are encoded by separate genes. The amino acid sequences for the bovine and human β 2 subunit are identical, as are the amino acid sequences for the bovine and human β 1 subunit. This evolutionary conservation suggests that the two β subunits have different roles in the signal transduction process

  8. Translation activity of chimeric ribosomes composed of Escherichia coli and Bacillus subtilis or Geobacillus stearothermophilus subunits

    Directory of Open Access Journals (Sweden)

    Sayaka Tsuji

    2017-07-01

    Full Text Available Ribosome composition, consisting of rRNA and ribosomal proteins, is highly conserved among a broad range of organisms. However, biochemical studies focusing on ribosomal subunit exchangeability between organisms remain limited. In this study, we show that chimeric ribosomes, composed of Escherichia coli and Bacillus subtilis or E. coli and Geobacillus stearothermophilus subunits, are active for β-galactosidase translation in a highly purified E. coli translation system. Activities of the chimeric ribosomes showed only a modest decrease when using E. coli 30 S subunits, indicating functional conservation of the 50 S subunit between these bacterial species.

  9. Specific radioimmunoassay of HCG and its α and β subunits: methods and results

    International Nuclear Information System (INIS)

    Reuter, A.M.; Schoonbrood, J.; Franchimont, P.

    1976-01-01

    To create antisera that are specific for the radioimmunoassay of HCG and its subunits, the antisera are neutralized by incubation with LH or HCG. For each RIA system the inhibition curves of HCG and its subunits LH, FSH, TSH and STH are obtained. The 125 I labelled hormones HCG, α and β subunits and LH were chromatographed over a Sephadex G 100 column. Serum of menopausal and pregnant women were chromatographed in the same way and the fractions subjected to RIA. HCG and its subunits were determined by RIA in the sera of patients with different kinds of cancer

  10. Immunochemical analysis of Micrococcus lysodeikticus (luteus) F1-ATPase and its subunits.

    Science.gov (United States)

    Urban, C; Salton, M R

    1983-08-31

    The F1-ATPase from Micrococcus lysodeikticus has been purified to 95% protein homogeneity in this laboratory and as all other bacterial F1S, possesses five distinct subunits with molecular weights ranging from 60 000 to 10 000 (Huberman, M. and Salton, M.R.J. (1979) Biochim. Biophys. Acta 547, 230-240). In this communication, we demonstrate the immunochemical reactivities of antibodies to native and SDS-dissociated subunits with the native and dissociated F1-ATPase and show that: (1) the antibodies generated to the native or SDS-dissociated subunits react with the native molecule; (2) all of the subunits comprising the F1 are antigenically unique as determined by crossed immunoelectrophoresis and the Ouchterlony double-diffusion techniques; (3) antibodies to the SDS-denatured individual delta- and epsilon-subunits can be used to destabilize the interaction of these specific subunits with the rest of the native F1; and (4) all subunit antibodies as well as anti-native F1 were found to inhibit ATPase activity to varying degrees, the strongest inhibition being seen with antibodies to the total F1 and anti-alpha- and anti-beta-subunit antibodies. The interaction of specific subunit antibodies may provide a new and novel way to study further and characterize the catalytic portions of F1-ATPases and in general may offer an additional method for the examination of multimeric proteins.

  11. Determination of hCG-alpha subunit in threatened pregnancy

    International Nuclear Information System (INIS)

    Talas, M.; Pohanka, J.; Fingerova, H.; Janouskova, M.; Krikal, Z.; Prasilova, J.; Zupkova, H.

    1987-01-01

    Radioimmunoassay of the hCG-alpha subunit was made using an antibody anti hCG-alpha serum, highly purified hCG-alpha for 125 I-labelling and the standard hCG-alpha. Sera of healthy pregnant women sampled throughout the whole pregnancies were used to determine x-bar±S.D. of hCG-alpha for 14-day intervals. Included in the study were groups of women with high risk of premature labor, late toxemia of pregnancy, twins and fetal hypotrophy. It was shown that increased hCG-alpha is found in pregnant women in whom signs of late toxemia of pregnancy are combined with high risk of premature labor, or with twin pregnancies, while in those with fetal hypotrophy hCG-alpha is within normal limits. (author). 3 figs., 7 refs

  12. Chaperonin Structure - The Large Multi-Subunit Protein Complex

    Directory of Open Access Journals (Sweden)

    Irena Roterman

    2009-03-01

    Full Text Available The multi sub-unit protein structure representing the chaperonins group is analyzed with respect to its hydrophobicity distribution. The proteins of this group assist protein folding supported by ATP. The specific axial symmetry GroEL structure (two rings of seven units stacked back to back - 524 aa each and the GroES (single ring of seven units - 97 aa each polypeptide chains are analyzed using the hydrophobicity distribution expressed as excess/deficiency all over the molecule to search for structure-to-function relationships. The empirically observed distribution of hydrophobic residues is confronted with the theoretical one representing the idealized hydrophobic core with hydrophilic residues exposure on the surface. The observed discrepancy between these two distributions seems to be aim-oriented, determining the structure-to-function relation. The hydrophobic force field structure generated by the chaperonin capsule is presented. Its possible influence on substrate folding is suggested.

  13. Glycine Receptor α2 Subunit Activation Promotes Cortical Interneuron Migration

    Directory of Open Access Journals (Sweden)

    Ariel Avila

    2013-08-01

    Full Text Available Glycine receptors (GlyRs are detected in the developing CNS before synaptogenesis, but their function remains elusive. This study demonstrates that functional GlyRs are expressed by embryonic cortical interneurons in vivo. Furthermore, genetic disruption of these receptors leads to interneuron migration defects. We discovered that extrasynaptic activation of GlyRs containing the α2 subunit in cortical interneurons by endogenous glycine activates voltage-gated calcium channels and promotes calcium influx, which further modulates actomyosin contractility to fine-tune nuclear translocation during migration. Taken together, our data highlight the molecular events triggered by GlyR α2 activation that control cortical tangential migration during embryogenesis.

  14. The subunit structure of the extracellular hemoglobin of Biomphalaria glabrata

    International Nuclear Information System (INIS)

    Arndt, Marcio H.L.; Naves, Cristiani F.; Xavier, Luciana P.; Santoro, Marcelo M.

    1997-01-01

    Full text. The hemoglobin of Biomphalaria glabrata was purified to homogeneity by a two step purification protocol using a gel filtration column (Superose 6 HR/Pharmacia ) followed by an anion exchange chromatography (MONO-Q Sepharose/Pharmacia). The dissociation products were analysed by a 5 - 15 % Polyacrylamide gel electrophoresis containing Sodium Dodecyl Sulfate (SDS-PAGE) giving a band of 270 K Daltons and a band of 180 K Daltons after reduction with β-mercaptoethanol. The same profile was obtained in a 3.5 % Agarose gel electrophoresis containing SDS (SDS-AGE) showing additional bands of higher molecular weight. These bands were proposed to be monomers, dimers and trimers and, after reduction in a Bidimensional SDS-AGE, the proposed monomers and dimers were decomposed in two and four bands that were interpreted as 1 - 4 chains. The hemoglobin was digested by four different proteases ( Thrombin, Trypsin, Chymotrypsin and Subtilisin ) showing several equivalent fragments with molecular weights multiples of its minimum molecular weight ( 17.7 K Daltons). The circular dichroism spectrum of the protein showed a characteristic high α-helix content. We proposed that this hemoglobin is a pentamer of approx. 360 K Daltons subunits each formed by two 180 K Daltons chains linked in pairs by disulfide bridges and each of these chains comprises ten Heme binding domains. These data were compared to other Planorbidae extracellular hemoglobins. Up to now, the quaternary structure of this hemoglobin (shape and disposition of the subunits) is unknown. It is intended to elucidate its structure by Small Angle X-Ray Scattering in Brazilian National Laboratory of Synchrotron Light (LNLS). (author)

  15. Differential regulation of thyrotropin subunit apoprotein and carbohydrate biosynthesis by thyroid hormone

    International Nuclear Information System (INIS)

    Taylor, T.; Weintraub, B.D.

    1985-01-01

    The regulation of TSH apoprotein and carbohydrate biosynthesis by thyroid hormone was studied by incubating pituitaries from normal and hypothyroid (3 weeks post-thyroidectomy) rats in medium containing [ 14 C]alanine and [ 3 H] glucosamine. After 6 h, samples were sequentially treated with anti-TSH beta to precipitate TSH and free TSH beta, anti-LH beta to clear the sample of LH and free LH beta, then anti-LH alpha to precipitate free alpha-subunit. Total proteins were acid precipitated. All precipitates were subjected to electrophoresis on sodium dodecyl sulfate-polyacrylamide gels, which were then sliced and assayed by scintillation spectrometry. In hypothyroid pituitaries plus medium, [ 14 C]alanine incorporation in combined and free beta-subunits was 26 times normal and considerably greater than the 3.4-fold increase seen in total protein; combined and free alpha-subunits showed no specific increase in apoprotein synthesis. [ 3 H]Glucosamine incorporation in combined alpha- and beta-subunits in hypothyroid samples was 13 and 21 times normal, respectively, and was greater than the 1.9-fold increase in total protein; free alpha-subunit showed no specific increase in carbohydrate synthesis. The glucosamine to alanine ratio, reflecting relative glycosylation of newly synthesized molecules, was increased in hypothyroidism for combined alpha-subunits, but not for combined beta-subunits, free alpha-subunits, or total proteins. In summary, short term hypothyroidism selectively stimulated TSH beta apoprotein synthesis and carbohydrate synthesis of combined alpha- and beta-subunits. Hypothyroidism also increased the relative glycosylation of combined alpha-subunit. Thus, thyroid hormone deficiency appears to alter the rate-limiting step in TSH assembly (i.e. beta-subunit synthesis) as well as the carbohydrate structure of TSH, which may play important roles in its biological function

  16. Copolymer semiconductors comprising thiazolothiazole or benzobisthiazole, or benzobisoxazole electron acceptor subunits, and electron donor subunits, and their uses in transistors and solar cells

    Science.gov (United States)

    Jenekhe, Samson A; Subramaniyan, Selvam; Ahmed, Eilaf; Xin, Hao; Kim, Felix Sunjoo

    2014-10-28

    The inventions disclosed, described, and/or claimed herein relate to copolymers comprising copolymers comprising electron accepting A subunits that comprise thiazolothiazole, benzobisthiazole, or benzobisoxazoles rings, and electron donating subunits that comprise certain heterocyclic groups. The copolymers are useful for manufacturing organic electronic devices, including transistors and solar cells. The invention also relates to certain synthetic precursors of the copolymers. Methods for making the copolymers and the derivative electronic devices are also described.

  17. Roles of the β subunit hinge domain in ATP synthase F1 sector: Hydrophobic network formed by introduced βPhe174 inhibits subunit rotation

    International Nuclear Information System (INIS)

    Nakanishi-Matsui, Mayumi; Kashiwagi, Sachiko; Kojima, Masaki; Nonaka, Takamasa; Futai, Masamitsu

    2010-01-01

    The ATP synthase β subunit hinge domain (βPhe148 ∼ βGly186, P-loop/α-helixB/loop/β-sheet4, Escherichia coli residue numbering) dramatically changes in conformation upon nucleotide binding. We previously reported that F 1 with the βSer174 to Phe mutation in the domain lowered the γ subunit rotation speed, and thus decreased the ATPase activity [M. Nakanishi-Matsui, S. Kashiwagi, T. Ubukata, A. Iwamoto-Kihara, Y. Wada, M. Futai, Rotational catalysis of Escherichia coli ATP synthase F 1 sector. Stochastic fluctuation and a key domain of the β subunit, J. Biol. Chem. 282 (2007) 20698-20704.]. Homology modeling indicates that the amino acid replacement induces a hydrophobic network, in which the βMet159, βIle163, and βAla167 residues of the β subunit are involved together with the mutant βPhe174. The network is expected to stabilize the conformation of β DP (nucleotide-bound form of the β subunit), resulting in increased activation energy for transition to β E (empty β subunit). The modeling further predicts that replacement of βMet159 with Ala or Ile weakens the hydrophobic network. As expected, these two mutations experimentally suppressed the ATPase activities as well as subunit rotation of βS174F. Furthermore, the rotation rate decreased with the increase of the strength in the hydrophobic network. These results indicate that the smooth conformational change of the β subunit hinge domain is pertinent for the rotational catalysis.

  18. Regulated appearance of NMDA receptor subunits and channel functions during in vitro neuronal differentiation

    NARCIS (Netherlands)

    Jelitai, Márta; Schlett, Katalin; Varju, Patrícia; Eisel, Ulrich; Madarász, Emília

    The schedule of NMDA receptor subunit expression and the appearance of functional NMDA-gated ion channels were investigated during the retinoic acid (RA) induced neuronal differentiation of NE-4C, a p53-deficient mouse neuroectodermal progenitor cell line. NR2A. NR2B, and NR2D subunit transcripts

  19. Differential expression of BK channel isoforms and beta-subunits in rat neuro-vascular tissues

    DEFF Research Database (Denmark)

    Poulsen, Asser Nyander; Wulf, Helle; Hay-Schmidt, Anders

    2009-01-01

    We investigated the expression of splice variants and beta-subunits of the BK channel (big conductance Ca(2+)-activated K(+) channel, Slo1, MaxiK, K(Ca)1.1) in rat cerebral blood vessels, meninges, trigeminal ganglion among other tissues. An alpha-subunit splice variant X1(+24) was found expresse...

  20. Differential antibiotic sensitivity determined by the large ribosomal subunit in thermophilic archaea.

    OpenAIRE

    Ruggero, D; Londei, P

    1996-01-01

    Hybrid ribosomes obtained by mixing the ribosomal subunits of the extremely thermophilic archaea Sulfolobus solfataricus and Desulfurococcus mobilis were tested for their sensitivity to selected antibiotics. It is shown that structural differences in the large ribosomal subunits determine qualitatively and quantitatively the patterns of response to alpha-sarcin and paromomycin in these species.

  1. Isolation and characterization of recombinant human casein kinase II subunits alpha and beta from bacteria

    DEFF Research Database (Denmark)

    Grankowski, N; Boldyreff, B; Issinger, O G

    1991-01-01

    cDNA encoding the casein kinase II (CKII) subunits alpha and beta of human origin were expressed in Escherichia coli using expression vector pT7-7. Significant expression was obtained with E. coli BL21(DE3). The CKII subunits accounted for approximately 30% of the bacterial protein; however, most...

  2. Regulation of KV channel voltage-dependent activation by transmembrane β subunits

    Directory of Open Access Journals (Sweden)

    Xiaohui eSun

    2012-04-01

    Full Text Available Voltage-activated K+ (KV channels are important for shaping action potentials and maintaining resting membrane potential in excitable cells. KV channels contain a central pore-gate domain (PGD surrounded by four voltage-sensing domains (VSD. The VSDs will change conformation in response to alterations of the membrane potential thereby inducing the opening of the PGD. Many KV channels are heteromeric protein complexes containing auxiliary β subunits. These β subunits modulate channel expression and activity to increase functional diversity and render tissue specific phenotypes. This review focuses on the KV β subunits that contain transmembrane (TM segments including the KCNE family and the β subunits of large conductance, Ca2+- and voltage-activated K+ (BK channels. These TM β subunits affect the voltage-dependent activation of KV α subunits. Experimental and computational studies have described the structural location of these β subunits in the channel complexes and the biophysical effects on VSD activation, PGD opening and VSD-PGD coupling. These results reveal some common characteristics and mechanistic insights into KV channel modulation by TM β subunits.

  3. A-Raf kinase is a new interacting partner of protein kinase CK2 beta subunit

    DEFF Research Database (Denmark)

    Boldyreff, B; Issinger, O G

    1997-01-01

    In a search for protein kinase CK2 beta subunit binding proteins using the two-hybrid system, more than 1000 positive clones were isolated. Beside clones for the alpha' and beta subunit of CK2, there were clones coding for a so far unknown protein, whose partial cDNA sequence was already deposited...

  4. Purification of the alpha and beta subunits of phosphorylase kinase for structural studies

    International Nuclear Information System (INIS)

    Sotiroudis, T.G.; Heilmeyer, L.M.G. Jr.; Crabb, J.W.

    1987-01-01

    Structural analysis of the alpha (Mr, 132,000) and beta (Mr, 113,000) subunits of phosphorylase kinase may provide clues to their yet unknown functions however purification remains problematic. Preparative RP-HPLC procedures yield inconveniently large, dilute solutions and concentration steps are required prior to subunit modification and fragmentation. Concentration of the β subunit usually results in significant losses due to insolubility. Using preparative SDS-polyacrylamide gel electrophoresis, they have purified the α, 7 , and β subunits from rabbit muscle phosphorylase kinase in a soluble and concentrated form suitable for structural studies. Phosphorylase kinase labelled with fluorescein isothiocyanate in the α and α' subunits and fully 14 C-S-carboxymethylated was fractionated on a 5% acrylamide Laemmli slab gel. The subunit bands were visualized by fluorescence and by SDS precipitation then excised and electroeluted in the presence of SDS using an ELUTRAP device. From 4.5 mg of enzyme applied to a 4.5 mm thick gel about 70% of the α subunit and about 90% of the β subunit were typically recovered in less than 1 ml with overnight elution

  5. Submitochondrial distributions and stabilities of subunits 4, 5, and 6 of yeast cytochrome oxidase in assembly defective mutants.

    Science.gov (United States)

    Glerum, D M; Tzagoloff, A

    1997-08-04

    The concentration and submitochondrial distribution of the subunit polypeptides of cytochrome oxidase have been studied in wild type yeast and in different mutants impaired in assembly of this respiratory complex. All the subunit polypeptides of the enzyme are associated with mitochondrial membranes of wild type cells, except for a small fraction of subunits 4 and 6 that is recovered in the soluble protein fraction of mitochondria. Cytochrome oxidase mutants consistently display a severe reduction in the steady-state concentration of subunit 1 due to its increased turnover. As a consequence, most of subunit 4, which normally is associated with subunit 1, is found in the soluble fraction. A similar shift from membrane-bound to soluble subunit 6 is seen in mutants blocked in expression of subunit 5a. In contrast, null mutations in COX6 coding for subunit 6 promote loss of subunit 5a. The absence of subunit 5a in the cox6 mutant is the result of proteolytic degradation rather than regulation of its expression by subunit 6. The possible role of the ATP-dependent proteases Rca1p and Afg3p in proteolysis of subunits 1 and 5a has been assessed in strains with combined mutations in COX6, RCA1, and/or AFG3. Immunochemical assays indicate that another protease(s) must be responsible for most of the proteolytic loss of these proteins.

  6. Persistence of the mitochondrial permeability transition in the absence of subunit c of human ATP synthase.

    Science.gov (United States)

    He, Jiuya; Ford, Holly C; Carroll, Joe; Ding, Shujing; Fearnley, Ian M; Walker, John E

    2017-03-28

    The permeability transition in human mitochondria refers to the opening of a nonspecific channel, known as the permeability transition pore (PTP), in the inner membrane. Opening can be triggered by calcium ions, leading to swelling of the organelle, disruption of the inner membrane, and ATP synthesis, followed by cell death. Recent proposals suggest that the pore is associated with the ATP synthase complex and specifically with the ring of c-subunits that constitute the membrane domain of the enzyme's rotor. The c-subunit is produced from three nuclear genes, ATP5G1 , ATP5G2 , and ATP5G3 , encoding identical copies of the mature protein with different mitochondrial-targeting sequences that are removed during their import into the organelle. To investigate the involvement of the c-subunit in the PTP, we generated a clonal cell, HAP1-A12, from near-haploid human cells, in which ATP5G1 , ATP5G2 , and ATP5G3 were disrupted. The HAP1-A12 cells are incapable of producing the c-subunit, but they preserve the characteristic properties of the PTP. Therefore, the c-subunit does not provide the PTP. The mitochondria in HAP1-A12 cells assemble a vestigial ATP synthase, with intact F 1 -catalytic and peripheral stalk domains and the supernumerary subunits e, f, and g, but lacking membrane subunits ATP6 and ATP8. The same vestigial complex plus associated c-subunits was characterized from human 143B ρ 0 cells, which cannot make the subunits ATP6 and ATP8, but retain the PTP. Therefore, none of the membrane subunits of the ATP synthase that are involved directly in transmembrane proton translocation is involved in forming the PTP.

  7. Mining Protein Evolution for Insights into Mechanisms of Voltage-Dependent Sodium Channel Auxiliary Subunits.

    Science.gov (United States)

    Molinarolo, Steven; Granata, Daniele; Carnevale, Vincenzo; Ahern, Christopher A

    2018-02-21

    Voltage-gated sodium channel (VGSC) beta (β) subunits have been called the "overachieving" auxiliary ion channel subunit. Indeed, these subunits regulate the trafficking of the sodium channel complex at the plasma membrane and simultaneously tune the voltage-dependent properties of the pore-forming alpha-subunit. It is now known that VGSC β-subunits are capable of similar modulation of multiple isoforms of related voltage-gated potassium channels, suggesting that their abilities extend into the broader voltage-gated channels. The gene family for these single transmembrane immunoglobulin beta-fold proteins extends well beyond the traditional VGSC β1-β4 subunit designation, with deep roots into the cell adhesion protein family and myelin-related proteins - where inherited mutations result in a myriad of electrical signaling disorders. Yet, very little is known about how VGSC β-subunits support protein trafficking pathways, the basis for their modulation of voltage-dependent gating, and, ultimately, their role in shaping neuronal excitability. An evolutionary approach can be useful in yielding new clues to such functions as it provides an unbiased assessment of protein residues, folds, and functions. An approach is described here which indicates the greater emergence of the modern β-subunits roughly 400 million years ago in the early neurons of Bilateria and bony fish, and the unexpected presence of distant homologues in bacteriophages. Recent structural breakthroughs containing α and β eukaryotic sodium channels containing subunits suggest a novel role for a highly conserved polar contact that occurs within the transmembrane segments. Overall, a mixture of approaches will ultimately advance our understanding of the mechanism for β-subunit interactions with voltage-sensor containing ion channels and membrane proteins.

  8. Topographic antigenic determinants recognized by monoclonal antibodies on human choriogonadotropin beta-subunit

    International Nuclear Information System (INIS)

    Bidart, J.M.; Troalen, F.; Salesse, R.; Bousfield, G.R.; Bohuon, C.J.; Bellet, D.H.

    1987-01-01

    We describe a first attempt to study the antibody-combining sites recognized by monoclonal antibodies raised against the beta-subunit of human choriogonadotropin (hCG). Two groups of antibodies were first defined by their ability to recognize only the free beta-subunit or the free and combined subunit. Antibodies FBT-11 and FBT-11-L bind only to hCG beta-subunit but not to hCG, whereas antibodies FBT-10 and D1E8 bind to both the beta-subunit and the hormone. In both cases, the antigenic determinants were localized to the core of the protein (residues 1-112), indicating the weak immunogenicity of the specific carboxyl-terminal extension of hCG-beta. Nine synthetic peptides spanning different regions of hCG-beta and lutropin-beta were assessed for their capacity to inhibit antibody binding. A synthetic peptide inclusive of the NH2-terminal region (residues 1-7) of the hCG beta-subunit was found to inhibit binding to the radiolabeled subunit of a monoclonal antibody specific for free hCG-beta (FBT-11). Further delineation of the antigenic site recognized by this antibody provided evidence for the involvement of fragment 82-92. Moreover, monoclonal antibody FBT-11 inhibited the recombination of hCG-beta to hCG-alpha, indicating that its antigenic determinant might be located nearby or in the hCG-beta portion interacting with the alpha-subunit. Binding of monoclonal antibody FBT-10, corresponding to the second antigenic determinant, was weakly inhibited by fragment 82-105 and did not impair the recombination of the hCG beta-subunit to the hCG alpha-subunit. Its combining site appeared to be located in a region of the intact native choriogonadotropin present at the surface of the hormone-receptor complex

  9. Subunit association as the stabilizing determinant for archaeal methionine adenosyltransferases.

    Science.gov (United States)

    Garrido, Francisco; Alfonso, Carlos; Taylor, John C; Markham, George D; Pajares, María A

    2009-07-01

    Archaea contain a class of methionine adenosyltransferases (MATs) that exhibit substantially higher stability than their mesophilic counterparts. Their sequences are highly divergent, but preserve the essential active site motifs of the family. We have investigated the origin of this increased stability using chemical denaturation experiments on Methanococcus jannaschii MAT (Mj-MAT) and mutants containing single tryptophans in place of tyrosine residues. The results from fluorescence, circular dichroism, hydrodynamic, and enzyme activity measurements showed that the higher stability of Mj-MAT derives largely from a tighter association of its subunits in the dimer. Local fluorescence changes, interpreted using secondary structure predictions, further identify the least stable structural elements as the C-terminal ends of beta-strands E2 and E6, and the N-terminus of E3. Dimer dissociation however requires a wider perturbation of the molecule. Additional analysis was initially hindered by the lack of crystal structures for archaeal MATs, a limitation that we overcame by construction of a 3D-homology model of Mj-MAT. This model predicts preservation of the chain topology and three-domain organization typical of this family, locates the least stable structural elements at the flat contact surface between monomers, and shows that alterations in all three domains are required for dimer dissociation.

  10. The Regulation of NF-κB Subunits by Phosphorylation

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    Frank Christian

    2016-03-01

    Full Text Available The NF-κB transcription factor is the master regulator of the inflammatory response and is essential for the homeostasis of the immune system. NF-κB regulates the transcription of genes that control inflammation, immune cell development, cell cycle, proliferation, and cell death. The fundamental role that NF-κB plays in key physiological processes makes it an important factor in determining health and disease. The importance of NF-κB in tissue homeostasis and immunity has frustrated therapeutic approaches aimed at inhibiting NF-κB activation. However, significant research efforts have revealed the crucial contribution of NF-κB phosphorylation to controlling NF-κB directed transactivation. Importantly, NF-κB phosphorylation controls transcription in a gene-specific manner, offering new opportunities to selectively target NF-κB for therapeutic benefit. This review will focus on the phosphorylation of the NF-κB subunits and the impact on NF-κB function.

  11. DNA binding properties of the small cascade subunit Csa5.

    Directory of Open Access Journals (Sweden)

    Michael Daume

    Full Text Available CRISPR-Cas systems provide immunity against viral attacks in archaeal and bacterial cells. Type I systems employ a Cas protein complex termed Cascade, which utilizes small CRISPR RNAs to detect and degrade the exogenic DNA. A small sequence motif, the PAM, marks the foreign substrates. Previously, a recombinant type I-A Cascade complex from the archaeon Thermoproteus tenax was shown to target and degrade DNA in vitro, dependent on a native PAM sequence. Here, we present the biochemical analysis of the small subunit, Csa5, of this Cascade complex. T. tenax Csa5 preferentially bound ssDNA and mutants that showed decreased ssDNA-binding and reduced Cascade-mediated DNA cleavage were identified. Csa5 oligomerization prevented DNA binding. Specific recognition of the PAM sequence was not observed. Phylogenetic analyses identified Csa5 as a universal member of type I-A systems and revealed three distinct groups. A potential role of Csa5 in R-loop stabilization is discussed.

  12. Editing modifies the GABA(A) receptor subunit alpha3

    DEFF Research Database (Denmark)

    Ohlson, Johan; Pedersen, Jakob Skou; Haussler, David

    2007-01-01

    Adenosine to inosine (A-to-I) pre-mRNA editing by the ADAR enzyme family has the potential to increase the variety of the proteome. This editing by adenosine deamination is essential in mammals for a functional brain. To detect novel substrates for A-to-I editing we have used an experimental method...... to find selectively edited sites and combined it with bioinformatic techniques that find stem-loop structures suitable for editing. We present here the first verified editing candidate detected by this screening procedure. We show that Gabra-3, which codes for the alpha3 subunit of the GABA(A) receptor......, is a substrate for editing by both ADAR1 and ADAR2. Editing of the Gabra-3 mRNA recodes an isoleucine to a methionine. The extent of editing is low at birth but increases with age, reaching close to 100% in the adult brain. We therefore propose that editing of the Gabra-3 mRNA is important for normal brain...

  13. Vaccine profile of herpes zoster (HZ/su) subunit vaccine.

    Science.gov (United States)

    Cunningham, Anthony L; Heineman, Thomas

    2017-07-01

    Herpes zoster (HZ) causes an often severe and painful rash in older people and may be complicated by prolonged pain (postherpetic neuralgia; PHN) and by dissemination in immune-compromised patients. HZ results from reactivation of latent varicella-zoster virus (VZV) infection, often associated with age-related or other causes of decreased T cell immunity. A live attenuated vaccine boosts this immunity and provides partial protection against HZ, but this decreases with age and declines over 8 years. Areas covered: A new HZ subunit (HZ/su) vaccine combines a key surface VZV glycoprotein (E) with a T cell-boosting adjuvant system (AS01 B ) and is administered by two intramuscular injections two months apart. Expert commentary: HZ/su showed excellent efficacy of ~90% in immunocompetent adults ≥50 and ≥70 years of age, respectively, in the ZOE-50 and ZOE-70 phase III controlled trials. Efficacy was unaffected by advancing age and persisted for >3 years. Approximately 9.5% of subjects had severe, but transient (1-2 days) injection site pain, swelling or redness. Compliance with both vaccine doses was high (95%). The vaccine will have a major impact on HZ management. Phase I-II trials showed safety and immunogenicity in severely immunocompromised patients. Phase III trial results are expected soon.

  14. Design of a hyperstable 60-subunit protein icosahedron

    Science.gov (United States)

    Hsia, Yang; Bale, Jacob B.; Gonen, Shane; Shi, Dan; Sheffler, William; Fong, Kimberly K.; Nattermann, Una; Xu, Chunfu; Huang, Po-Ssu; Ravichandran, Rashmi; Yi, Sue; Davis, Trisha N.; Gonen, Tamir; King, Neil P.; Baker, David

    2016-07-01

    The icosahedron is the largest of the Platonic solids, and icosahedral protein structures are widely used in biological systems for packaging and transport. There has been considerable interest in repurposing such structures for applications ranging from targeted delivery to multivalent immunogen presentation. The ability to design proteins that self-assemble into precisely specified, highly ordered icosahedral structures would open the door to a new generation of protein containers with properties custom-tailored to specific applications. Here we describe the computational design of a 25-nanometre icosahedral nanocage that self-assembles from trimeric protein building blocks. The designed protein was produced in Escherichia coli, and found by electron microscopy to assemble into a homogenous population of icosahedral particles nearly identical to the design model. The particles are stable in 6.7 molar guanidine hydrochloride at up to 80 degrees Celsius, and undergo extremely abrupt, but reversible, disassembly between 2 molar and 2.25 molar guanidinium thiocyanate. The icosahedron is robust to genetic fusions: one or two copies of green fluorescent protein (GFP) can be fused to each of the 60 subunits to create highly fluorescent ‘standard candles’ for use in light microscopy, and a designed protein pentamer can be placed in the centre of each of the 20 pentameric faces to modulate the size of the entrance/exit channels of the cage. Such robust and customizable nanocages should have considerable utility in targeted drug delivery, vaccine design and synthetic biology.

  15. Fc receptor gamma subunit polymorphisms and systemic lupus erythematosus

    International Nuclear Information System (INIS)

    Al-Ansari, Aliya; Ollier, W.E.; Gonzalez-Gay, Miguel A.; Gul, Ahmet; Inanac, Murat; Ordi, Jose; Teh, Lee-Suan; Hajeer, Ali H.

    2004-01-01

    To investigate the possible association between Fc receptor gamma polymorphisms and systemic lupus erythematosus (SLE). We have investigated the full FcR gamma gene for polymorphisms using polymerase chain reaction (PCR)-single strand confirmational polymorphisms and DNA sequencing .The polymorphisms identified were genotype using PCR-restriction fragment length polymorphism. Systemic lupus erythematosus cases and controls were available from 3 ethnic groups: Turkish, Spanish and Caucasian. The study was conducted in the year 2001 at the Arthritis Research Campaign, Epidemiology Unit, Manchester University Medical School, Manchester, United Kingdom. Five single nucleotide polymorphisms were identified, 2 in the promoter, one in intron 4 and, 2 in the 3'UTR. Four of the 5 single nucleotide polymorphisms (SNPs) were relatively common and investigated in the 3 populations. Allele and genotype frequencies of all 4 investigated SNPs were not statistically different cases and controls. fc receptor gamma gene does not appear to contribute to SLE susceptibility. The identified polymorphisms may be useful in investigating other diseases where receptors containing the FcR gamma subunit contribute to the pathology. (author)

  16. Crystal structure of the P pilus rod subunit PapA.

    Directory of Open Access Journals (Sweden)

    Denis Verger

    2007-05-01

    Full Text Available P pili are important adhesive fibres involved in kidney infection by uropathogenic Escherichia coli strains. P pili are assembled by the conserved chaperone-usher pathway, which involves the PapD chaperone and the PapC usher. During pilus assembly, subunits are incorporated into the growing fiber via the donor-strand exchange (DSE mechanism, whereby the chaperone's G1 beta-strand that complements the incomplete immunoglobulin-fold of each subunit is displaced by the N-terminal extension (Nte of an incoming subunit. P pili comprise a helical rod, a tip fibrillum, and an adhesin at the distal end. PapA is the rod subunit and is assembled into a superhelical right-handed structure. Here, we have solved the structure of a ternary complex of PapD bound to PapA through donor-strand complementation, itself bound to another PapA subunit through DSE. This structure provides insight into the structural basis of the DSE reaction involving this important pilus subunit. Using gel filtration chromatography and electron microscopy on a number of PapA Nte mutants, we establish that PapA differs in its mode of assembly compared with other Pap subunits, involving a much larger Nte that encompasses not only the DSE region of the Nte but also the region N-terminal to it.

  17. Structural characterization of recombinant crustacyanin subunits from the lobster Homarus americanus

    International Nuclear Information System (INIS)

    Ferrari, Michele; Folli, Claudia; Pincolini, Elisa; McClintock, Timothy S.; Rössle, Manfred; Berni, Rodolfo; Cianci, Michele

    2012-01-01

    The two recombinant apo subunits H1 and H2 from H. americanus have been structurally characterized. Reconstitution studies with astaxanthin reproduced the bathochromic shift of 85–95 nm typical of the natural crustacyanin subunits. Crustacean crustacyanin proteins are linked to the production and modification of carapace colour, with direct implications for fitness and survival. Here, the structural and functional properties of the two recombinant crustacyanin subunits H 1 and H 2 from the American lobster Homarus americanus are reported. The two subunits are structurally highly similar to the corresponding natural apo crustacyanin CRTC and CRTA subunits from the European lobster H. gammarus. Reconstitution studies of the recombinant crustacyanin proteins H 1 and H 2 with astaxanthin reproduced the bathochromic shift of 85–95 nm typical of the natural crustacyanin subunits from H. gammarus in complex with astaxanthin. Moreover, correlations between the presence of crustacyanin genes in crustacean species and the resulting carapace colours with the spectral properties of the subunits in complex with astaxanthin confirmed this genotype–phenotype linkage

  18. Self-subunit swapping occurs in another gene type of cobalt nitrile hydratase.

    Directory of Open Access Journals (Sweden)

    Yi Liu

    Full Text Available Self-subunit swapping is one of the post-translational maturation of the cobalt-containing nitrile hydratase (Co-NHase family of enzymes. All of these NHases possess a gene organization of , which allows the activator protein to easily form a mediatory complex with the α-subunit of the NHase after translation. Here, we discovered that the incorporation of cobalt into another type of Co-NHase, with a gene organization of , was also dependent on self-subunit swapping. We successfully isolated a recombinant NHase activator protein (P14K of Pseudomonas putida NRRL-18668 by adding a Strep-tag N-terminal to the P14K gene. P14K was found to form a complex [α(StrepP14K(2] with the α-subunit of the NHase. The incorporation of cobalt into the NHase of P. putida was confirmed to be dependent on the α-subunit substitution between the cobalt-containing α(StrepP14K(2 and the cobalt-free NHase. Cobalt was inserted into cobalt-free α(StrepP14K(2 but not into cobalt-free NHase, suggesting that P14K functions not only as a self-subunit swapping chaperone but also as a metallochaperone. In addition, NHase from P. putida was also expressed by a mutant gene that was designed with a order. Our findings expand the general features of self-subunit swapping maturation.

  19. Voltage-Gated Sodium Channel β1/β1B Subunits Regulate Cardiac Physiology and Pathophysiology

    Directory of Open Access Journals (Sweden)

    Nnamdi Edokobi

    2018-04-01

    Full Text Available Cardiac myocyte contraction is initiated by a set of intricately orchestrated electrical impulses, collectively known as action potentials (APs. Voltage-gated sodium channels (NaVs are responsible for the upstroke and propagation of APs in excitable cells, including cardiomyocytes. NaVs consist of a single, pore-forming α subunit and two different β subunits. The β subunits are multifunctional cell adhesion molecules and channel modulators that have cell type and subcellular domain specific functional effects. Variants in SCN1B, the gene encoding the Nav-β1 and -β1B subunits, are linked to atrial and ventricular arrhythmias, e.g., Brugada syndrome, as well as to the early infantile epileptic encephalopathy Dravet syndrome, all of which put patients at risk for sudden death. Evidence over the past two decades has demonstrated that Nav-β1/β1B subunits play critical roles in cardiac myocyte physiology, in which they regulate tetrodotoxin-resistant and -sensitive sodium currents, potassium currents, and calcium handling, and that Nav-β1/β1B subunit dysfunction generates substrates for arrhythmias. This review will highlight the role of Nav-β1/β1B subunits in cardiac physiology and pathophysiology.

  20. Three human alcohol dehydrogenase subunits: cDNA structure and molecular and evolutionary divergence

    International Nuclear Information System (INIS)

    Ikuta, T.; Szeto, S.; Yoshida, A.

    1986-01-01

    Class I human alcohol dehydrogenase (ADH; alcohol:NAD + oxidoreductase, EC 1.1.1.1) consists of several homo- and heterodimers of α, β, and γ subunits that are governed by the ADH1, ADH2, and ADH3 loci. The authors previously cloned a full length of cDNA for the β subunit, and the complete sequence of 374 amino acid residues was established. cDNAs for the α and γ subunits were cloned and characterized. A human liver cDNA library, constructed in phage λgt11, was screened by using a synthetic oligonucleotide probe that was matched to the γ but not to the β sequence. Clone pUCADHγ21 and clone pUCADHα15L differed from β cDNA with respect to restriction sites and hybridization with the nucleotide probe. Clone pUCADHγ21 contained an insertion of 1.5 kilobase pairs (kbp) and encodes 374 amino acid residues compatible with the reported amino acid sequence of the γ subunit. Clone pUCADHα15L contained an insertion of 2.4 kbp and included nucleotide sequences that encode 374 amino acid residues for another subunit, the γ subunit. In addition, this clone contained the sequences that encode the COOH-terminal part of the β subunit at its extended 5' region. The amino acid sequences and coding regions of the cDNAs of the three subunits are very similar. A high degree of resemblance is observed also in their 3' noncoding regions. However, distinctive differences exist in the vicinity of the Zn-binding cysteine residue at position 46. Based on the cDNA sequences and the deduced amino acid sequences of the three subunits, their structural and evolutionary relationships are discussed