Science.gov

Sample records for high yielding mutants

  1. Root-determined hypernodulation mutant of Lotus japonicus shows high-yielding characteristics.

    PubMed

    Yokota, Keisuke; Li, Yong Yi; Hisatomi, Masahiro; Wang, Yanxu; Ishikawa, Kaori; Liu, Chi-Te; Suzuki, Shino; Aonuma, Kho; Aono, Toshihiro; Nakamoto, Tomomi; Oyaizu, Hiroshi

    2009-07-01

    Here we report the phenotypic characteristics of a novel hypernodulation mutant, Ljrdh1 (root-determined hypernodulation 1) of Lotus japonicus. At 12 weeks after rhizobial inoculation, there were no differences between the growth of Ljrdh1 and, wild-type. However, Ljrdh1 showed 2 to 3 times higher nitrogen-fixing activity, and seed and pod yields, were approximately 50% higher than the wild-type. This is the first report of a legume hypernodulation mutant showing normal growth and a high-yielding characteristic under optimal cultivation conditions.

  2. High yield antibiotic producing mutants of Streptomyces erythreus induced by low energy ion implantation

    NASA Astrophysics Data System (ADS)

    Yu, Chen; Zhixin, Lin; Zuyao, Zou; Feng, Zhang; Duo, Liu; Xianghuai, Liu; Jianzhong, Tang; Weimin, Zhu; Bo, Huang

    1998-05-01

    Conidia of Streptomyces erythreus, an industrial microbe, were implanted by nitrogen ions with energy of 40-60 keV and fluence from 1 × 10 11 to 5 × 10 14 ions/cm 2. The logarithm value of survival fraction had good linear relationship with the logarithm value of fluence. Some mutants with a high yield of erythromycin were induced by ion implantation. The yield increment was correlated with the implantation fluence. Compared with the mutation results induced by ultraviolet rays, mutation effects of ion implantation were obvious having higher increasing erythromycin potency and wider mutation spectrum. The spores of Bacillus subtilis were implanted by arsenic ions with energy of 100 keV. The distribution of implanted ions was measured by Rutherford Backscattering Spectrometry (RBS) and calculated in theory. The mechanism of mutation induced by ion implantation was discussed.

  3. High yield production of a mutant Nippostrongylus brasiliensis acetylcholinesterase in Pichia pastoris and its purification.

    PubMed

    Richter, Sven; Nieveler, Jens; Schulze, Holger; Bachmann, Till T; Schmid, Rolf D

    2006-04-01

    The mutant M301A of the acetylcholinesterase B from Nippostrongylus brasiliensis (NbAChE) was produced in a high-cell-density fermentation of a recombinant methylotrophic yeast Pichia pastoris. Dissolved oxygen (DO) spikes were used as an indicator for feeding the carbon source. Wet cell weight (WCW) reached after 8 days a maximum value of 316 g/L and the OD600 at this time was 280. The acetylcholinesterase activity increased up to 6,600 U/mL corresponding to an expression rate of 2 g of NbAChE per liter supernatant. The specific activity of the mutant NbAChE was determined after purification as 3,300 U/mg. Active site titration with chlorpyrifos, a strong AChE inhibitor, yielded in a specific activity of 3,400 U/mg. The enzyme was secreted by Pichia pastoris. Therefore, it could be concentrated from culture broth by cross-flow-filtration (50 kDa cut-off membrane). It was further purified in one-step anion-exchange chromatography, using a XK 50/20 column filled with 125 mL Q Sepharose HP. Mutant NbAChE was purified 1.9-fold up to a purity of 97% and a yield of 87%. The isolated enzyme was nearly homogenous, as seen on the silver stained SDS-PAGE as well as by a single peak after gel filtration. This extraordinary high expression rate and the ease of purification is an important prerequisite for their practical application, for example in biosensors for the detection of neurotoxic insecticides.

  4. Stability analysis of a high fibre yield and low lignin content "thick stem" mutant in tossa jute (Corchorus olitorius L.).

    PubMed

    Mandal, Aninda; Datta, Animesh K

    2014-01-01

    A "thick stem" mutant of Corchorus olitorius L. was induced at M2 (0.50%, 4 h, EMS) and the true breeding mutant is assessed across generations (M5 to M7) considering morphometric traits as well as SEM analysis of pollen grains and raw jute fibres, stem anatomy, cytogenetical attributes, and lignin content in relation to control. Furthermore, single fibre diameter and tensile strength are also analysed. The objective is to assess the stability of mutant for its effective exploration for raising a new plant type in tossa jute for commercial exploitation and efficient breeding. The mutant trait is monogenic recessive to normal. Results indicate that "thick stem" mutant is stable across generations (2n = 14) with distinctive high seed and fibre yield and significantly low lignin content. Stem anatomy of the mutant shows significant enhancement in fibre zone, number of fibre pyramids and fibre bundles per pyramid, and diameter of fibre cell in relation to control. Moreover, tensile strength of mutant fibre is significantly higher than control fibre and the trait is inversely related to fibre diameter. However the mutant is associated with low germination frequency, poor seed viability, and high pollen sterility, which may be eliminated through mutational approach followed by rigorous selection and efficient breeding.

  5. Stability analysis of a high fibre yield and low lignin content "thick stem" mutant in tossa jute (Corchorus olitorius L.).

    PubMed

    Mandal, Aninda; Datta, Animesh K

    2014-01-01

    A "thick stem" mutant of Corchorus olitorius L. was induced at M2 (0.50%, 4 h, EMS) and the true breeding mutant is assessed across generations (M5 to M7) considering morphometric traits as well as SEM analysis of pollen grains and raw jute fibres, stem anatomy, cytogenetical attributes, and lignin content in relation to control. Furthermore, single fibre diameter and tensile strength are also analysed. The objective is to assess the stability of mutant for its effective exploration for raising a new plant type in tossa jute for commercial exploitation and efficient breeding. The mutant trait is monogenic recessive to normal. Results indicate that "thick stem" mutant is stable across generations (2n = 14) with distinctive high seed and fibre yield and significantly low lignin content. Stem anatomy of the mutant shows significant enhancement in fibre zone, number of fibre pyramids and fibre bundles per pyramid, and diameter of fibre cell in relation to control. Moreover, tensile strength of mutant fibre is significantly higher than control fibre and the trait is inversely related to fibre diameter. However the mutant is associated with low germination frequency, poor seed viability, and high pollen sterility, which may be eliminated through mutational approach followed by rigorous selection and efficient breeding. PMID:24860822

  6. Clostridium beijerinckii mutant obtained by atmospheric pressure glow discharge producing high proportions of butanol and solvent yields.

    PubMed

    Guo, Ting; Tang, Yan; Xi, Yong-lan; He, Ai-yong; Sun, Bai-jun; Wu, Hao; Liang, Da-feng; Jiang, Min; Ouyang, Ping-kai

    2011-12-01

    With 30 g glucose/l as carbon source, Clostridium beijerinckii ART124, a mutant created by atmospheric pressure glow discharge, produced 13.7 g total solvent/l (containing 3.1 g acetone/l, 10.4 g butanol/l and 0.2 g ethanol/l) in 72 h. The mutant could also use sucrose or xylose or a mixture of glucose/xylose/arabinose with nearly equal yields.

  7. Directed breeding of an Arthrobacter mutant for high-yield production of cyclic adenosine monophosphate by N + ion implantation

    NASA Astrophysics Data System (ADS)

    Song, He; Chen, Xiaochun; Cao, Jiaming; Fang, Ting; Bai, Jianxin; Xiong, Jian; Ying, Hanjie

    2010-08-01

    To obtain a cyclic adenosine monophosphate (cAMP) high-yield production strain, Arthrobacter NG-1 was mutated by N + ion implantation with an energy level of 10 keV and dose of 7×10 15 ions/cm 2. Combined with directed screening methods, a xanthine-defective and 8-azaguanine (8-AG)-resistant mutant Arthrobacter A302 was selected. The concentration of cAMP produced by this mutant was 41.7% higher than that of the original strain and reached 9.78 g/L. Through ten-generation investigation, the capability of cAMP production of A302 was found to be stable. Compared with the original strain, the special activities of key enzymes in A302, which influenced the cAMP biosynthesis, was analyzed. IMP dehydrogenase activity was defective, whereas PRPP amidotransferase, sAMP synthetase and adenylate cyclase activities were increased by 61.5%, 147% and 21.7%, respecitively, which might explain the mutagenesis mechanism by N + ions implantation under the enzymatic level.

  8. Production and Screening of High Yield Avermectin B1b Mutant of Streptomyces avermitilis 41445 Through Mutagenesis

    PubMed Central

    Siddique, Samia; Syed, Quratulain; Adnan, Ahmad; Qureshi, Fahim Ashraf

    2014-01-01

    Background: Secondary metabolite production from wild strains is very low for economical purpose therefore certain strain improvement strategies are required to achieve hundred times greater yield of metabolites. Most important strain improvement techniques include physical and chemical mutagenesis. Broad spectrum mutagenesis through UV irradiation is the most important and convenient physical method. Objectives: The present study was conducted for enhanced production of avermectin B1b from Streptomyces avermitilis 41445 by mutagenesis using ultraviolet (UV) radiation, ethidium bromide (EB), and ethyl methanesulfonate (EMS) as mutagens. Materials and Methods: S. avermitilis DSM 41445 maintained on yeast extract malt extract glucose medium (YMG) was used as inoculum for SM2 fermentation medium. Spores of S. avermitilis DSM 41445 were exposed to UV radiation for physical broad spectrum mutagenesis and to EMS and EB for chemical mutagenesis. For each mutagen, the lethality rate and mutation rate were calculated along with positive mutation rate. Results: Avermectin B1b-hyper-producing mutant, produced using these three different methods, was selected according to the HPLC results. The mutant obtained after 45 minutes of UV radiation to the spores of S. avermitilis 41445, was found to be the best mutant for the enhanced production of avermectin B1b component (254.14 mg/L). Other avermectin B1b-hyper-producing mutants, were obtained from EMS (1 µL/mL) and EB (30 µL/mL) treatments, and yielded 202.63 mg/L and 199.30 mg/L of B1b, respectively. Conclusions: The hereditary stability analysis of the UV mentioning 45 minutes revealed the UV exposure time for mutants and 3 represented the colony taken from the plate irradiated for 45 minutes mutant showed that the production of avermectin B1b remained constant and no reverse mutation occurred after 15 generations. PMID:25147669

  9. Stability Analysis of a High Fibre Yield and Low Lignin Content “Thick Stem” Mutant in Tossa Jute (Corchorus olitorius L.)

    PubMed Central

    Mandal, Aninda; Datta, Animesh K.

    2014-01-01

    A “thick stem” mutant of Corchorus olitorius L. was induced at M2 (0.50%, 4 h, EMS) and the true breeding mutant is assessed across generations (M5 to M7) considering morphometric traits as well as SEM analysis of pollen grains and raw jute fibres, stem anatomy, cytogenetical attributes, and lignin content in relation to control. Furthermore, single fibre diameter and tensile strength are also analysed. The objective is to assess the stability of mutant for its effective exploration for raising a new plant type in tossa jute for commercial exploitation and efficient breeding. The mutant trait is monogenic recessive to normal. Results indicate that “thick stem” mutant is stable across generations (2n = 14) with distinctive high seed and fibre yield and significantly low lignin content. Stem anatomy of the mutant shows significant enhancement in fibre zone, number of fibre pyramids and fibre bundles per pyramid, and diameter of fibre cell in relation to control. Moreover, tensile strength of mutant fibre is significantly higher than control fibre and the trait is inversely related to fibre diameter. However the mutant is associated with low germination frequency, poor seed viability, and high pollen sterility, which may be eliminated through mutational approach followed by rigorous selection and efficient breeding. PMID:24860822

  10. Development and evaluation of rice giant embryo mutants for high oil content originated from a high-yielding cultivar ‘Mizuhochikara’

    PubMed Central

    Sakata, Mitsukazu; Seno, Mari; Matsusaka, Hiroaki; Takahashi, Kiyomi; Nakamura, Yuki; Yamagata, Yoshiyuki; Angeles, Enrique R.; Mochizuki, Toshihiro; Kumamaru, Toshihiro; Sato, Masao; Enomoto, Akiko; Tashiro, Kosuke; Kuhara, Satoru; Satoh, Hikaru; Yoshimura, Atsushi

    2016-01-01

    Rice bran oil is a byproduct of the milling of rice (Oryza sativa L.). It offers various health benefits and has a beneficial fatty acid composition. To increase the amount of rice bran as a sink for triacylglycerol (TAG), we developed and characterized new breeding materials with giant embryos. To induce mutants, we treated fertilized egg cells of the high-yielding cultivar ‘Mizuhochikara’ with N-methyl-N-nitrosourea (MNU). By screening M2 seeds, we isolated four giant embryo mutant lines. Genetic analysis revealed that the causative loci in lines MGE12 and MGE13 were allelic to giant embryo (ge) on chromosome 7, and had base changes in the causal gene Os07g0603700. On the other hand, the causative loci in lines MGE8 and MGE14 were not allelic to ge, and both were newly mapped on chromosome 3. The TAG contents of all four mutant lines increased relative to their wild type, ‘Mizuhochikara’. MGE13 was agronomically similar to ‘Mizuhochikara’ and would be useful for breeding for improved oil content. PMID:27436953

  11. Reduced mutant yield at high doses in the Salmonella/activation assay: the cause is not always toxicity.

    PubMed

    McGregor, D; Prentice, R D; McConville, M; Lee, Y J; Caspary, W J

    1984-01-01

    In the Salmonella/activation assay developed by Ames et al [1973, 1975] toxicity is not measured, though it is recognized by the loss of a cloudy appearance on the plate. One approach to the measurement of toxicity is described here and uses a microscope-linked automated colony counter to estimate the number of microcolonies formed by histidine auxotrophs that stop growing after the depletion of histidine. This technique was used to evaluate the effect of toxicity on the revertant count for 16 mutagens, most of which were chosen because, from previous experience, their dose-response curves manifested a maximum at an intermediate dose tested. One of the sixteen, 2-nitrofluorene, was not toxic up to the maximum dose tested. The relationship between mutation and toxicity for the remaining fifteen allowed them to be grouped into two categories: (1) compounds that induced decreases in survival at the same dose at which the number of mutants decreased, and (2) compounds that induced toxicity, but survival was reduced at dose levels higher than those required to reduce the number of mutants. Possible explanations for this reduction of mutant counts occurring with little apparent concomitant increase in toxicity are examined. These results may be significant for attempts to estimate mutagenic potency and, to a lesser extent, construct mathematical models of the Ames test. PMID:6381041

  12. Generation of a catR deficient mutant of P. putida KT2440 that produces cis, cis-muconate from benzoate at high rate and yield.

    PubMed

    van Duuren, J B J H; Wijte, D; Leprince, A; Karge, B; Puchałka, J; Wery, J; Dos Santos, V A P Martins; Eggink, G; Mars, A E

    2011-12-10

    Pseudomonas putida KT2440-JD1 was derived from P. putida KT2440 after N-methyl-N'-nitro-N-nitrosoguanidine (NTG)-mutagenesis and exposure to 3-fluorobenzoate (3-FB). The mutant was no longer able to grow using benzoate as a sole carbon source, but co-metabolized benzoate to cis, cis-muconate during growth on glucose, which accumulated in the growth medium. The specific production rate (q(pm)) was 0.18±0.03 g cis, cis-muconate/(g(DCW) h) in continuous cultures, and increased to 1.4 g cis, cis-muconate/(g(DCW) h) during wash-out cultivation. Transcriptome analysis showed that the cat operon was not induced in P. putida KT2440-JD1 in the presence of 5mM benzoate, due to a point mutation in the highly conserved DNA binding domain of the transcriptional regulator (catR) of the cat operon. The ben operon was highly expressed in the presence of benzoate in the mutant and its parental strain. This operon contains PP_3166 (catA2), which was shown to be a second catechol 1,2-dioxygenase besides catA. P. putida KT2440-JD1 is the first cis, cis-muconate-accumulating mutant that was characterized at the genetic level. The specific production rate achieved is at least eight times higher than those reported for other cis, cis-muconate-producing strains.

  13. Breeding for blast-disease-resistant and high-yield Thai jasmine rice (Oryza sativa L. cv. KDML 105) mutants using low-energy ion beams

    NASA Astrophysics Data System (ADS)

    Mahadtanapuk, S.; Teraarusiri, W.; Phanchaisri, B.; Yu, L. D.; Anuntalabhochai, S.

    2013-07-01

    Low-energy ion beam was applied on mutation induction for plant breeding of blast-disease-resistant Thai jasmine rice (Oryza sativa L. cv. KDML 105). Seeds of the wild-type rice were bombarded in vacuum by nitrogen ion beam at energy of 60-80 keV to a beam fluence range of 2 × 1016-2 × 1017 ions/cm2. The ion-bombarded rice seeds were grown in soil for 2 weeks as transplanted rice in plastic pots at 1 seedling/pot. The seedlings were then screened for blast resistance by Pyricularia grisea inoculation with 106 spores/ml concentrations. The blast-resistant rice mutant was planted up to F6 generation with the consistent phenotypic variation. The high percentage of the blast-disease-resistant rice was analyzed with DNA fingerprint. The HAT-RAPD (high annealing temperature-random amplified polymorphic DNA) marker revealed the modified polymorphism fragment presenting in the mutant compared with wild type (KDML 105). The cDNA fingerprints were investigated and the polymorphism fragment was subcloned into pGEM-T easy vector and then sequenced. The sequence of this fragment was compared with those already contained in the database, and the fragment was found to be related to the Spotted leaf protein 11 (Spl11).

  14. Ascorbate, added after irradiation, reduces the mutant yield and alters the spectrum of CD59- mutations in A(L) cells irradiated with high LET carbon ions

    NASA Technical Reports Server (NTRS)

    Ueno, Akiko; Vannais, Diane; Lenarczyk, Marek; Waldren, Charles A.; Chatterjee, A. (Principal Investigator)

    2002-01-01

    It has been reported that X-ray induced HPRT- mutation in cultured human cells is prevented by ascorbate added after irradiation. Mutation extinction is attributed to neutralization by ascorbate, of radiation-induced long-lived radicals (LLR) with half-lives of several hours. We here show that post-irradiation treatment with ascorbate (5 mM added 30 min after radiation) reduces, but does not eliminate, the induction of CD59- mutants in human-hamster hybrid A(L) cells exposed to high-LET carbon ions (LET of 100 KeV/microm). RibCys, [2(R,S)-D-ribo-1',2',3',4'-Tetrahydroxybutyl]-thiazolidene-4(R)-ca riboxylic acid] (4 mM) gave a similar but lesser effect. The lethality of the carbon ions was not altered by these chemicals. Preliminary data are presented that ascorbate also alters the spectrum of CD59- mutations induced by the carbon beam, mainly by reducing the incidence of small mutations and mutants displaying transmissible genomic instability (TGI), while large mutations are unaffected. Our results suggest that LLR are important in initiating TGI.

  15. Expression in high yield of pig alpha 1 beta 1 Na,K-ATPase and inactive mutants D369N and D807N in Saccharomyces cerevisiae.

    PubMed

    Pedersen, P A; Rasmussen, J H; Jøorgensen, P L

    1996-02-01

    Studies of structure-function relationships in Na,K-ATPase require high yield expression of inactive mutations in cells without endogenous Na,K-ATPase activity. In this work we developed a host/vector system for expression of fully active pig Na,K-ATPase as well as the inactive mutations D369N and D807N at high levels in Saccharomyces cerevisiae. The alpha 1- and beta 1-subunit cDNAs were inserted into a single 2-microns-based plasmid with a high and regulatable copy number and strong galactose-inducible promoters allowing for stoichiometric alterations of gene dosage. The protease-deficient host strain was engineered to express high levels of GAL4 transactivating protein, thereby causing a 10-fold increase in expression to 32,500 +/- 3,000 [3H]ouabain sites/cell. In one bioreactor run 150-200 g of yeast were produced with 54 +/- 5 micrograms of Na,K-pump protein/g of cells. Through purification in membrane bound form the activity of the recombinant Na,K-ATPase was increased to 42-50 pmol/mg of protein. The Na,K dependence of ATP hydrolysis and the molar activity (4,500-7,000 min-1) were close to those of native pig kidney Na,K-ATPase. Mutations to the phosphorylation site (D369N) or presumptive cation sites (D807N), both devoid of Na,K-ATPase activity, were expressed in the yeast membrane at the same alpha-subunit concentration and [3H]ouabain binding capacity as the wild type Na,K-ATPase. The high yield and absence of endogenous activity allowed assay of [3H]ATP binding at equilibrium, demonstrating a remarkable 18-fold increase in affinity for ATP in consequence of reducing the negative charge at the phosphorylation site (D369N).

  16. Fast-germinating low β-glucan mutants induced in barley with improved malting quality and yield.

    PubMed

    Molina-Cano, J L; de Togores, F R; Royo, C; Pérez, A

    1989-11-01

    Mutation breeding has been used to improve the speed of germination in the high-yielding spring barley variety Troubadour. Five mutants were selected which combined fast germination and good agronomic performance. Two of these mutants yielded significantly more than did Troubadour over eight environments, and showed a clear improvement in their malting quality through an increase in extract yield. The improvement in malting quality appeared to be due to a decrease in the β-glucan content, which seemed to enhance the germination speed and thus the starch degradation. The improvement in grain yield is postulated to be due to a better early growth caused by the enhanced germination speed. All the described changes could theoretically be explained by a single mutation event in each of the mutant genotypes, affecting the quantity of β-glucans present in the endosperm.

  17. PRODUCING HIGH CORN YIELDS.

    ERIC Educational Resources Information Center

    Illinois Univ., Urbana. Coll. of Agriculture.

    RESOURCE MATERIAL ON CORN PRODUCTION FOR HIGH SCHOOL VOCATIONAL AGRICULTURE AND ADULT FARMER CLASSES WAS DESIGNED BY A STATE LEVEL GROUP OF SUBJECT MATTER SPECIALISTS, TEACHER EDUCATORS, SUPERVISORS, AND TEACHERS TO HELP SOLVE PROBLEMS THAT CONFRONT CORN PRODUCERS AT PLANTING TIME. THE SUBJECT MATTER CONCERNS PLANTING TIME, DEPTH, ROW WIDTH,…

  18. Gradient domestication of Haematococcus pluvialis mutant with 15% CO2 to promote biomass growth and astaxanthin yield.

    PubMed

    Cheng, Jun; Li, Ke; Yang, Zongbo; Lu, Hongxiang; Zhou, Junhu; Cen, Kefa

    2016-09-01

    In order to increase biomass yield and reduce culture cost of Haematococcus pluvialis with flue gas from coal-fired power plants, a screened mutant by nuclear irradiation was gradually domesticated with 15% CO2 to promote biomass dry weight and astaxanthin yield. The biomass yield of mutant after 10 generations of 15% CO2 domestication increased to 1.3 times as that with air. With the optimization of nitrogen and phosphorus concentration, the biomass dry weight was further increased by 62%. The astaxanthin yield induced with 15% CO2 and high light of 135 μmol photons m(-2) s(-1) increased to 87.4mg/L, which was 6 times higher than that induced with high light in air.

  19. Gradient domestication of Haematococcus pluvialis mutant with 15% CO2 to promote biomass growth and astaxanthin yield.

    PubMed

    Cheng, Jun; Li, Ke; Yang, Zongbo; Lu, Hongxiang; Zhou, Junhu; Cen, Kefa

    2016-09-01

    In order to increase biomass yield and reduce culture cost of Haematococcus pluvialis with flue gas from coal-fired power plants, a screened mutant by nuclear irradiation was gradually domesticated with 15% CO2 to promote biomass dry weight and astaxanthin yield. The biomass yield of mutant after 10 generations of 15% CO2 domestication increased to 1.3 times as that with air. With the optimization of nitrogen and phosphorus concentration, the biomass dry weight was further increased by 62%. The astaxanthin yield induced with 15% CO2 and high light of 135 μmol photons m(-2) s(-1) increased to 87.4mg/L, which was 6 times higher than that induced with high light in air. PMID:27259189

  20. High Persister Mutants in Mycobacterium tuberculosis

    PubMed Central

    Torrey, Heather L.; Keren, Iris; Via, Laura E.; Lee, Jong Seok; Lewis, Kim

    2016-01-01

    Mycobacterium tuberculosis forms drug-tolerant persister cells that are the probable cause of its recalcitrance to antibiotic therapy. While genetically identical to the rest of the population, persisters are dormant, which protects them from killing by bactericidal antibiotics. The mechanism of persister formation in M. tuberculosis is not well understood. In this study, we selected for high persister (hip) mutants and characterized them by whole genome sequencing and transcriptome analysis. In parallel, we identified and characterized clinical isolates that naturally produce high levels of persisters. We compared the hip mutants obtained in vitro with clinical isolates to identify candidate persister genes. Genes involved in lipid biosynthesis, carbon metabolism, toxin-antitoxin systems, and transcriptional regulators were among those identified. We also found that clinical hip isolates exhibited greater ex vivo survival than the low persister isolates. Our data suggest that M. tuberculosis persister formation involves multiple pathways, and hip mutants may contribute to the recalcitrance of the infection. PMID:27176494

  1. High Yielding Microbubble Production Method

    PubMed Central

    Fiabane, Joe; Prentice, Paul; Pancholi, Ketan

    2016-01-01

    Microfluidic approaches to microbubble production are generally disadvantaged by low yield and high susceptibility to (micro)channel blockages. This paper presents an alternative method of producing microbubbles of 2.6 μm mean diameter at concentrations in excess of 30 × 106 mL−1. In this method, the nitrogen gas flowing inside the liquid jet is disintegrated into spray of microbubble when air surrounding this coflowing nitrogen gas-liquid jet passes through a 100 μm orifice at high velocity. Resulting microbubble foam has the polydispersity index of 16%. Moreover, a ratio of mean microbubble diameter to channel width ratio was found to be less than 0.025, which substantially alleviates the occurrence of blockages during production. PMID:27034935

  2. A high yield neutron target

    NASA Technical Reports Server (NTRS)

    Alger, D. L.; Steinberg, R.; Weisenbach, P.

    1974-01-01

    Target, in cylinder form, rotates rapidly in front of beam. Titanium tritide film is much thicker than range of accelerated deutron. Sputtering electrode permits full use of thick film. Stream of high-velocity coolant provides efficient transfer of heat from target.

  3. Escherichia coli mutants resistant to inactivation by high hydrostatic pressure.

    PubMed Central

    Hauben, K J; Bartlett, D H; Soontjens, C C; Cornelis, K; Wuytack, E Y; Michiels, C W

    1997-01-01

    Alternating cycles of exposure to high pressure and outgrowth of surviving populations were used to select for highly pressure-resistant mutants of Escherichia coli MG1655. Three barotolerant mutants (LMM1010, LMM1020, and LMM1030) were isolated independently by using outgrowth temperatures of 30, 37, and 42 degrees C, respectively. Survival of these mutants after pressure treatment for 15 min at ambient temperature was 40 to 85% at 220 MPa and 0.5 to 1.5% at 800 MPa, while survival of the parent strain, MG1655, decreased from 15% at 220 MPa to 2 x 10(-8)% at 700 MPa. Heat resistance of mutants LMM1020 and LMM1030 was also altered, as evident by higher D values at 58 and 60 degrees C and reduced z values compared to those for the parent strain. D and z values for mutant LMM1010 were not significantly different from those for the parent strain. Pressure sensitivity of the mutants increased from 10 to 50 degrees C, as opposed to the parent strain, which showed a minimum around 40 degrees C. The ability of the mutants to grow at moderately elevated pressure (50 MPa) was reduced at temperatures above 37 degrees C, indicating that resistance to pressure inactivation is unrelated to barotolerant growth. The development of high levels of barotolerance as demonstrated in this work should cause concern about the safety of high-pressure food processing. PMID:9055412

  4. High yield fabrication of fluorescent nanodiamonds

    PubMed Central

    Boudou, Jean-Paul; Curmi, Patrick; Jelezko, Fedor; Wrachtrup, Joerg; Aubert, Pascal; Sennour, Mohamed; Balasubramanian, Gopalakrischnan; Reuter, Rolf; Thorel, Alain; Gaffet, Eric

    2009-01-01

    A new fabrication method to produce homogeneously fluorescent nanodiamonds with high yields is described. The powder obtained by high energy ball milling of fluorescent high pressure, high temperature diamond microcrystals was converted in a pure concentrated aqueous colloidal dispersion of highly crystalline ultrasmall nanoparticles with a mean size less than or equal to 10 nm. The whole fabrication yield of colloidal quasi-spherical nanodiamonds was several orders of magnitude higher than those previously reported starting from microdiamonds. The results open up avenues for the industrial cost-effective production of fluorescent nanodiamonds with well-controlled properties. PMID:19451687

  5. Yield strength of molybdenum at high pressures.

    PubMed

    Jing, Qiumin; Bi, Yan; Wu, Qiang; Jing, Fuqian; Wang, Zhigang; Xu, Jian; Jiang, Sheng

    2007-07-01

    In the diamond anvil cell technology, the pressure gradient approach is one of the three major methods in determining the yield strength for various materials at high pressures. In the present work, by in situ measuring the thickness of the sample foil, we have improved the traditional technique in this method. Based on this modification, the yield strength of molybdenum at pressures has been measured. Our main experimental conclusions are as follows: (1) The measured yield strength data for three samples with different initial thickness (100, 250, and 500 microm) are in good agreement above a peak pressure of 10 GPa. (2) The measured yield strength can be fitted into a linear formula Y=0.48(+/-0.19)+0.14(+/-0.01)P (Y and P denote the yield strength and local pressure, respectively, both of them are in gigapascals) in the local pressure range of 8-21 GPa. This result is in good agreement with both Y=0.46+0.13P determined in the pressure range of 5-24 GPa measured by the radial x-ray diffraction technique and the previous shock wave data below 10 GPa. (3) The zero-pressure yield strength of Mo is 0.5 GPa when we extrapolate our experimental data into the ambient pressure. It is close to the tensile strength of 0.7 GPa determined by Bridgman [Phys. Rev. 48, 825 (1934)] previously. The modified method described in this article therefore provides the confidence in determination of the yield strength at high pressures. PMID:17672772

  6. Determination of essential fatty acid composition among mutant lines of Canola (Brassica napus), through high pressure liquid chromatography.

    PubMed

    Raza, Ghulam; Siddique, Aquil; Khan, Imtiaz Ahmad; Ashraf, Muhammed Yasin; Khatri, Abdullah

    2009-12-01

    The present study aimed to quantify the methyl esters of lenoleic acid (LA), gamma-lenolenic acid (LNA) and oleic acid (OL) in the oil of Brassica napus mutants. Five stable mutants (ROO-75/1, ROO-100/6, ROO-125/12, ROO-125/14, and ROO-125/17) of B. napus cv. 'Rainbow' (P) and three mutants (W97-95/16, W97-0.75/11 and W97-.075/13) of B. napus cv. 'Westar' (P) at M6 stage, exhibiting better yield and yield components, were analyzed for essential fatty acids. The highest seed yield was observed in the mutant (ROO-100/6) followed by ROO-125/14 of Rainbow, that is, 34% and 32% higher than their parent plants, respectively. Westar mutant W97-75/11 also showed 30% higher seed yield than its parent plant. High performance liquid chromatography analysis of the composition of fatty acids indicated that OL was the most dominant fatty acid, ranging from 39.1 to 66.3%; LA was second (15.3-41.6%) and LNA was third (18.1-28.9%). Mutant ROO-125/14 showed higher OL contents than parent (Rainbow). These results are expected to support the approval of ROO-125/14 in the National Uniform Varietal Yield Trials (NUVYT) as a new variety based on high oil quality.

  7. Development and characterisation of highly antibiotic resistant Bartonella bacilliformis mutants

    PubMed Central

    Gomes, Cláudia; Martínez-Puchol, Sandra; Ruiz-Roldán, Lidia; Pons, Maria J.; del Valle Mendoza, Juana; Ruiz, Joaquim

    2016-01-01

    The objective was to develop and characterise in vitro Bartonella bacilliformis antibiotic resistant mutants. Three B. bacilliformis strains were plated 35 or 40 times with azithromycin, chloramphenicol, ciprofloxacin or rifampicin discs. Resistance-stability was assessed performing 5 serial passages without antibiotic pressure. MICs were determined with/without Phe-Arg-β-Napthylamide and artesunate. Target alterations were screened in the 23S rRNA, rplD, rplV, gyrA, gyrB, parC, parE and rpoB genes. Chloramphenicol and ciprofloxacin resistance were the most difficult and easiest (>37.3 and 10.6 passages) to be selected, respectively. All mutants but one selected with chloramphenicol achieved high resistance levels. All rifampicin, one azithromycin and one ciprofloxacin mutants did not totally revert when cultured without antibiotic pressure. Azithromycin resistance was related to L4 substitutions Gln-66 → Lys or Gly-70 → Arg; L4 deletion Δ62–65 (Lys-Met-Tyr-Lys) or L22 insertion 83::Val-Ser-Glu-Ala-His-Val-Gly-Lys-Ser; in two chloramphenicol-resistant mutants the 23S rRNA mutation G2372A was detected. GyrA Ala-91 → Val and Asp-95 → Gly and GyrB Glu474 → Lys were detected in ciprofloxacin-resistant mutants. RpoB substitutions Gln-527 → Arg, His-540 → Tyr and Ser-545 → Phe plus Ser-588 → Tyr were detected in rifampicin-resistant mutants. In 5 mutants the effect of efflux pumps on resistance was observed. Antibiotic resistance was mainly related to target mutations and overexpression of efflux pumps, which might underlie microbiological failures during treatments. PMID:27667026

  8. Pre-thymic somatic mutation leads to high mutant frequency at hypoxanthine-guanine phosphoribosyltransferase gene

    SciTech Connect

    Jett, J.

    1994-12-01

    While characterizing the background mutation spectrum of the Hypoxathine-guanine phosphoribosyltransferase (HPRT) gene in a healthy population, an outlier with a high mutant frequency of thioguanine resistant lymphocytes was found. When studied at the age of 46, this individual had been smoking 60 cigarettes per day for 38 years. His mutant frequency was calculated at 3.6 and 4.2x10{sup {minus}4} for two sampling periods eight months apart. Sequencing analysis of the HPRT gene in his mutant thioguanine resistant T lymphocytes was done to find whether the cells had a high rate of mutation, or if the mutation was due to a single occurrence of mutation and, if so, when in the T lymphocyte development the mutation occurred. By T-cell receptor analysis it has been found that out of 35 thioguanine resistant clones there was no dominant gamma T cell receptor gene rearrangement. During my appointment in the Science & Engineering Research Semester, I found that 34 of those clones have the same base substitution of G{yields}T at cDNA position 197. Due to the consistent mutant frequency from both sampling periods and the varying T cell receptors, the high mutant frequency cannot be due to recent proliferation of a mature mutant T lymphocyte. From the TCR and DNA sequence analysis we conclude that the G{yields}T mutation must have occurred in a T lymphocyte precursor before thymic differentiation so that the thioguanine resistant clones share the same base substitution but not the same gamma T cell receptor gene.

  9. Distributions of enzyme residues yielding mutants with improved substrate specificities from two different directed evolution strategies.

    PubMed

    Paramesvaran, Janahan; Hibbert, Edward G; Russell, Andrew J; Dalby, Paul A

    2009-07-01

    A previous study of random mutations, mostly introduced by error-prone PCR (EPPCR) or DNA shuffling (DS), demonstrated that those closer to the enzyme active site were more effective than distant ones at improving enzyme activity, substrate specificity or enantioselectivity. Since then, many studies have taken advantage of this observation by targeting site-directed saturation mutagenesis (SDSM) to residues closer to or within enzyme active sites. Here, we have analysed a set of SDSM studies, in parallel to a similar set from EPPCR/DS, to determine whether the greater range of amino-acid types accessible by SDSM affects the distances at which the most effective sites occur. We have also analysed the relative effectiveness for obtaining beneficial mutants of residues with different degrees of natural sequence variation, as determined by their sequence entropy which is related to sequence conservation. These analyses attempt to answer the question-how well focused have targeted mutagenesis strategies been? We also compared two different sets of active-site atoms from which to measure distances and found that the inclusion of catalytic, substrate and cofactor atoms refined the analysis compared to using a single key catalytic atom. Using this definition, we found that EPPCR/DS is not effective for altering substrate specificity at sites that are within 5 A of the active-site atoms. In contrast, SDSM is most effective when targeted to residues at <5-6 A from the catalytic, substrate or cofactor atom, and also for residues with intermediate sequence entropies. Furthermore, SDSM is capable of altering substrate specificity at highly and completely conserved residues in the active site. The results suggest ways in which directed evolution by SDSM could be improved for greater efficiency in terms of reducing the library sizes required to obtain beneficial mutations that alter substrate specificity.

  10. Hyperproinsulinemia in a three-generation Caucasian family due to mutant proinsulin (Arg{sup 65}{yields}His) not associated with impaired glucose tolerance: The contribution of mutant proinsulin to insulin bioactivity

    SciTech Connect

    Roder, M.E.; Vissing, H.; Nauck, M.A.

    1996-04-01

    Familial hyperproinsulinemia is a genetic abnormality characterized by an increased proportion of proinsulin immunoreactivity in the circulation due to mutations affecting the posttranslational processing of proinsulin. In affected Japanese families, this has been associated with noninsulin-dependent diabetes mellitus or impaired glucose tolerance. A three-generation Caucasian family with hyperproinsulinemia was identified through unexplained hyperinsulinemia in a normal volunteer participating in a metabolic study. High pressure liquid chromatography analysis of fasting plasma revealed a major peak eluting close to the position of proinsulin. Direct sequencing of the proinsulin gene exon 3 showed a heterozygous point mutation (CGT{yields}CAT) resulting in the substitution of Arg{yields}His in position 65 (corresponding to the AC cleavage site) in the index case, his mother, and his maternal grandmother. All affected subjects had normal oral glucose tolerance. In the basal state and after oral glucose administration, their proinsulin responses were slightly reduced. However, when calculating insulin bioactivity by assuming 9% activity for mutant Arg{sup 65}{yields}His proinsulin, responses in affected subjects were comparable to those in normal subjects. In conclusion, our data demonstrate hyperproinsulinemia in a three-generation Caucasian family due to heterozygous mutant Arg{sup 65}{yields}His proinsulin. This was not associated with impaired glucose tolerance. These results suggest that this mutation in the heterozygous state per se does not affect glucose tolerance and that the biological activity of mutant proinsulin contributes to glucose homeostasis in this family. The association of the same mutation with impaired glucose tolerance or diabetes in previous studies may be the result of selection bias or associated conditions (e.g. the genetic background of the kindreds examined). 29 refs., 5 figs., 3 tabs.

  11. Development of high-yield influenza A virus vaccine viruses

    PubMed Central

    Ping, Jihui; Lopes, Tiago J.S.; Nidom, Chairul A.; Ghedin, Elodie; Macken, Catherine A.; Fitch, Adam; Imai, Masaki; Maher, Eileen A.; Neumann, Gabriele; Kawaoka, Yoshihiro

    2015-01-01

    Vaccination is one of the most cost-effective ways to prevent infection. Influenza vaccines propagated in cultured cells are approved for use in humans, but their yields are often suboptimal. Here, we screened A/Puerto Rico/8/34 (PR8) virus mutant libraries to develop vaccine backbones (defined here as the six viral RNA segments not encoding haemagglutinin and neuraminidase) that support high yield in cell culture. We also tested mutations in the coding and regulatory regions of the virus, and chimeric haemagglutinin and neuraminidase genes. A combination of high-yield mutations from these screens led to a PR8 backbone that improved the titres of H1N1, H3N2, H5N1 and H7N9 vaccine viruses in African green monkey kidney and Madin–Darby canine kidney cells. This PR8 backbone also improves titres in embryonated chicken eggs, a common propagation system for influenza viruses. This PR8 vaccine backbone thus represents an advance in seasonal and pandemic influenza vaccine development. PMID:26334134

  12. Integrated process for high conversion and high yield protein PEGylation.

    PubMed

    Pfister, David; Morbidelli, Massimo

    2016-08-01

    Over the past decades, PEGylation has become a powerful technique to increase the in vivo circulation half-life of therapeutic proteins while maintaining their activity. The development of new therapeutic proteins is likely to require further improvement of the PEGylation methods to reach even better selectivity and yield for reduced costs. The intensification of the PEGylation process was investigated through the integration of a chromatographic step in order to increase yield and conversion for the production of mono-PEGylated protein. Lysozyme was used as a model protein to demonstrate the feasibility of such approach. In the integrated reaction/separation process, chromatography was used as fractionation technique in order to isolate and recycle the unreacted protein from the PEGylated products. This allows operating the reactor with short reaction times so as to minimize the production of multi-PEGylated proteins (i.e., conjugated to more than one polymer). That is, the reaction is stopped before the desired product (i.e., the mono-PEGylated protein) can further react, thus leading to limited conversion but high yield. The recycling of the unreacted protein was then considered to drive the protein overall conversion to completion. This approach has great potential to improve processes whose yield is limited by the further reaction of the product leading to undesirable by-products. Biotechnol. Bioeng. 2016;113: 1711-1718. © 2016 Wiley Periodicals, Inc.

  13. Mutant selection of Hahella chejuensis KCTC 2396 and statistical optimization of medium components for prodigiosin yield-up.

    PubMed

    Kim, Sung Jin; Lee, Hong Kum; Lee, Yoo Kyung; Yim, Joung Han

    2008-04-01

    Prodigiosin is a natural red pigment with algicidal activity against Cochlodinium polykrikoides, a major harmful red-tide microalga. To increase the yield of prodigiosin, a mutant of Hahella chejuenesis KCTC 2396, assigned M3349, was developed by an antibiotic mutagenesis using chloramphenicol. When cultured in Sucrose-based Marine Broth medium (SMB), M3349 could produce prodigiosin at 1.628+/-0.06 g/L, while wild type producing at 0.658+/-0.12 g/L under the same conditions. To increase the yield of prodigiosin production by M3349, significant medium components were determined using a two-level Plackett-Burman statistical design technique. Among fourteen components included in SMB medium, NaCl, Na2SiO3, MgCl2, H3BO3, Na2HPO4, Na2SO4, and CaCl2 were determined to be important for prodigiosin production. The medium formulation was finally optimized using a Box-Behnken design as follows: sucrose 10.0, peptone 8.0, yeast extract 2.0, NaCl 10.0, Na2SO4 12.0, CaCl2 1.8, MgCl2 0.7 g/L; and H3BO3 22.0, Na2HPO4 20.0, Na2SiO3 8.0 mg/L. The predicted maximum yield of prodigiosin in the optimized medium was 2.43 g/L by the Box-Behnken design, while the practical production was 2.60+/-0.176 g/L, which was 3.9 times higher than wild type with SMB Medium (0.658 g/L).

  14. Bacteriophage-insensitive mutants for high quality Crescenza manufacture

    PubMed Central

    Chirico, Donatella; Gorla, Arianna; Verga, Viola; Pedersen, Per D.; Polgatti, Eliseo; Cava, Antonio; Dal Bello, Fabio

    2014-01-01

    Streptococcus thermophilus is a thermophilic lactic acid bacterium used as starter culture for the manufacture of fermented dairy products. For the production of Crescenza and other soft cheeses, Sacco has developed and provides dairies with three different defined blends of S. thermophilus strains. Each blend contains two different S. thermophilus strains. The strains were selected based on their unique technological properties as well as different phage profiles. Analysis of 133 whey samples collected in 2009–2010 from Italian dairies showed a high prevalence (about 50%) of bacteriophage attacks on the blend ST020. More specifically, the strain S. thermophilus ST1A was found to be the preferred target of the bacteriophages. A bacteriophage insensitive mutant (BIM5) of the phage-sensitive strain ST1A was successfully developed and used to substitute strain ST1A in the Crescenza starter culture ST020. The strain BIM5 showed identical technological and industrial traits as those of the phage-sensitive strain ST1A. The improved resistance of the modified Crescenza starter culture ST020R was confirmed at Italian dairies, and its effectiveness monitored on 122 whey samples collected in 2011–2012. Compared to the previous values (2009–2010), the use of the phage-hardened blend ST020R allowed reducing of frequency of phage attacks from about 50 to less than 5% of the whey samples investigated. PMID:24834065

  15. Identification of rice ethylene-response mutants and characterization of MHZ7/OsEIN2 in distinct ethylene response and yield trait regulation.

    PubMed

    Ma, Biao; He, Si-Jie; Duan, Kai-Xuan; Yin, Cui-Cui; Chen, Hui; Yang, Chao; Xiong, Qing; Song, Qing-Xin; Lu, Xiang; Chen, Hao-Wei; Zhang, Wan-Ke; Lu, Tie-Gang; Chen, Shou-Yi; Zhang, Jin-Song

    2013-11-01

    Ethylene plays essential roles in adaptive growth of rice plants in water-saturating environment; however, ethylene signaling pathway in rice is largely unclear. In this study, we report identification and characterization of ethylene-response mutants based on the specific ethylene-response phenotypes of etiolated rice seedlings, including ethylene-inhibited root growth and ethylene-promoted coleoptile elongation, which is different from the ethylene triple-response phenotype in Arabidopsis. We establish an efficient system for screening and a set of rice mutants have been identified. Genetic analysis reveals that these mutants form eight complementation groups. All the mutants show insensitivity or reduced sensitivity to ethylene in root growth but exhibit differential responses in coleoptile growth. One mutant group mhz7 has insensitivity to ethylene in both root and coleoptile growth. We identified the corresponding gene by a map-based cloning method. MHZ7 encodes a membrane protein homologous to EIN2, a central component of ethylene signaling in Arabidopsis. Upon ethylene treatment, etiolated MHZ7-overexpressing seedlings exhibit enhanced coleoptile elongation, increased mesocotyl growth and extremely twisted short roots, featuring enhanced ethylene-response phenotypes in rice. Grain length was promoted in MHZ7-transgenic plants and 1000-grain weight was reduced in mhz7 mutants. Leaf senescent process was also affected by MHZ7 expression. Manipulation of ethylene signaling may improve adaptive growth and yield-related traits in rice.

  16. The yield and post-yield behavior of high-density polyethylene

    NASA Technical Reports Server (NTRS)

    Semeliss, M. A.; Wong, R.; Tuttle, M. E.

    1990-01-01

    An experimental and analytical evaluation was made of the yield and post-yield behavior of high-density polyethylene, a semi-crystalline thermoplastic. Polyethylene was selected for study because it is very inexpensive and readily available in the form of thin-walled tubes. Thin-walled tubular specimens were subjected to axial loads and internal pressures, such that the specimens were subjected to a known biaxial loading. A constant octahederal shear stress rate was imposed during all tests. The measured yield and post-yield behavior was compared with predictions based on both isotropic and anisotropic models. Of particular interest was whether inelastic behavior was sensitive to the hydrostatic stress level. The major achievements and conclusions reached are discussed.

  17. Engineering of Corynebacterium glutamicum for high-yield L-valine production under oxygen deprivation conditions.

    PubMed

    Hasegawa, Satoshi; Suda, Masako; Uematsu, Kimio; Natsuma, Yumi; Hiraga, Kazumi; Jojima, Toru; Inui, Masayuki; Yukawa, Hideaki

    2013-02-01

    We previously demonstrated efficient L-valine production by metabolically engineered Corynebacterium glutamicum under oxygen deprivation. To achieve the high productivity, a NADH/NADPH cofactor imbalance during the synthesis of l-valine was overcome by engineering NAD-preferring mutant acetohydroxy acid isomeroreductase (AHAIR) and using NAD-specific leucine dehydrogenase from Lysinibacillus sphaericus. Lactate as a by-product was largely eliminated by disrupting the lactate dehydrogenase gene ldhA. Nonetheless, a few other by-products, particularly succinate, were still produced and acted to suppress the L-valine yield. Eliminating these by-products therefore was deemed key to improving theL-valine yield. By additionally disrupting the phosphoenolpyruvate carboxylase gene ppc, succinate production was effectively suppressed, but both glucose consumption and L-valine production dropped considerably due to the severely elevated intracellular NADH/NAD(+) ratio. In contrast, this perturbed intracellular redox state was more than compensated for by deletion of three genes associated with NADH-producing acetate synthesis and overexpression of five glycolytic genes, including gapA, encoding NADH-inhibited glyceraldehyde-3-phosphate dehydrogenase. Inserting feedback-resistant mutant acetohydroxy acid synthase and NAD-preferring mutant AHAIR in the chromosome resulted in higher L-valine yield and productivity. Deleting the alanine transaminase gene avtA suppressed alanine production. The resultant strain produced 1,280 mM L-valine at a yield of 88% mol mol of glucose(-1) after 24 h under oxygen deprivation, a vastly improved yield over our previous best.

  18. Synthesis, purification, and characterization of an Arg sub 152 yields Glu site-directed mutant of recombinant human blood clotting factor VII

    SciTech Connect

    Wildgoose, P.; Kisiel, W. ); Berkner, K.L. )

    1990-04-03

    Coagulation factor VII circulates in blood as a single-chain zymogen of a serine protease and is converted to its activated two-chain form, factor VIIa, by cleavage of an internal peptide bond located at Arg{sub 152}-Ile{sub 153}. Previous studies using serine protease active-site inhibitors suggest that zymogen factor VII may possess sufficient proteolytic activity to initiate the extrinsic pathway of blood coagulation. In order to assess the putative intrinsic proteolytic activity of single-chain factor VII, the authors have constructed a site-specific mutant of recombinant human factor VII in which arginine-152 has been replaced with a glutamic acid residue. Mutant factor VII was purified in a single step from culture supernatants of baby hamster kidney cells transfected with a plasmid containing the sequence for Arg{sub 152} {yields} Glu factor VII using a calcium-dependent, murine anti-factor VII monoclonal antibody column. The clotting activity of mutant factor VII was completely inhibited following incubation with dansyl-Glu-Gly-Arg chloromethyl ketone, suggesting that the apparent clotting activity of mutant factor VII was due to a contaminating serine protease. Immunoblots of mutant factor VII with human factor IXa revealed no cleavage, whereas incubation of mutant factor VII with human factor Xa resulted in cleavage of mutant factor VII and the formation of a lower molecular weight degradation product migrating at M{sup r}{approx}40 000. The results are consistent with the proposal that zymogen factor VII possesses no intrinsic proteolytic activity toward factor X or factor IX.

  19. Determining yields in high solids enzymatic hydrolysis of biomass.

    PubMed

    Kristensen, Jan B; Felby, Claus; Jørgensen, Henning

    2009-05-01

    As technologies for utilizing biomass for fuel and chemical production continue to improve, enzymatic hydrolysis can be run at still higher solids concentrations. For hydrolyses that initially contain little or no free water (10-40% total solids, w/w), the saccharification of insoluble polymers into soluble sugars involves changes of volume, density, and proportion of insoluble solids. This poses a new challenge when determining the degree of hydrolysis (conversion yield). Experiments have shown that calculating the yield from the resulting sugar concentration in the supernatant of the slurry and using the assumed initial volume leads to significant overestimations of the yield. By measuring the proportion of insoluble solids in the slurry as well as the sugar concentration and specific gravity of the aqueous phase, it is possible to precisely calculate the degree of conversion. The discrepancies between the different ways of calculating yields are demonstrated along with a nonlaborious method for approximating yields in high solids hydrolysis. PMID:18836690

  20. [Evaluation of penicillin expandase mutants and complex substrate inhibition characteristics at high concentrations of penicillin G].

    PubMed

    Wu, Linjun; Fan, Keqiang; Ji, Junjie; Yang, Keqian

    2015-12-01

    Penicillin expandase, also known as deacetoxycephalosporin C synthase (DAOCS), is an essential enzyme involved in cephalosporin C biosynthesis. To evaluate the catalytic behaviors of penicillin expandase under high penicillin G concentration and to identify mutants suitable for industrial applications, the specific activities of wild-type DAOCS and several mutants with increased activities toward penicillin G were determined by HPLC under high penicillin G concentrations. Their specific activity profiles were compared with theoretical predictions by different catalytic dynamics models. We evaluated the specific activities of wild-type DAOCS and previous reported high-activity mutants H4, H5, H6 and H7 at concentrations ranging from 5.6 to 500 mmol/L penicillin G. The specific activities of wild-type DAOCS and mutant H4 increased as penicillin G concentration increased, but decreased when concentrations of substrate go above 200 mmol/L. Other mutants H5, H6 and H7 showed more complex behaviors under high concentration of penicillin G. Among all tested enzymes, mutant H6 showed the highest activity when concentration of penicillin G is above 100 mmol/L. Our results revealed that the substrate inhibition to wild-type DAOCS' by penicillin G is noncompetitive. Other DAOCS mutants showed more complex trends in their specific activities at high concentration of penicillin G (>100 mmol/L), indicating more complex substrate inhibition mechanism might exist. The substrate inhibition and activity of DAOCS mutants at high penicillin G concentration provide important insight to help select proper mutants for industrial application. PMID:27093832

  1. High pressure intensification of cassava resistant starch (RS3) yields.

    PubMed

    Lertwanawatana, Proyphon; Frazier, Richard A; Niranjan, Keshavan

    2015-08-15

    Cassava starch, typically, has resistant starch type 3 (RS3) content of 2.4%. This paper shows that the RS3 yields can be substantially enhanced by debranching cassava starch using pullulanase followed by high pressure or cyclic high-pressure annealing. RS3 yield of 41.3% was obtained when annealing was carried out at 400MPa/60°C for 15 min, whereas it took nearly 8h to obtain the same yield under conventional atmospheric annealing at 60°C. The yield of RS3 could be further significantly increased by annealing under 400 MPa/60°C pressure for 15 min followed by resting at atmospheric pressure for 3h 45 min, and repeating this cycle for up to six times. Microstructural surface analysis of the product under a scanning electron microscope showed an increasingly rigid density of the crystalline structure formed, confirming higher RS3 content.

  2. HIGH TEMPERATURE STRESS ON FLORAL DEVELOPMENT AND YIELD OF COTTON

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Because a number of reproductive processes must occur in highly concerted fashion during the progamic phase (from pollination to fertilization) for successful fertilization and seed production to occur, final yield in cotton is exceptionally sensitive to high temperatures during the flowering period...

  3. Creation of High-Yield Polyhydroxyalkanoates Engineered Strains by Low Energy Ion Implantation

    NASA Astrophysics Data System (ADS)

    Qian, Shiquan; Cheng, Ying; Zhu, Suwen; Cheng, Beijiu

    2008-12-01

    Polyhydroxyalkanoates (PHAs), as a candidate for biodegradable plastic materials, can be synthesized by numerous microorganisms. However, as its production cost is high in comparison with those of chemically synthesized plastics, a lot of research has been focused on the efficient production of PHAs using different methods. In the present study, the mutation effects of PHAs production in strain pCB4 were investigated with implantation of low energy ions. It was found that under the implantation conditions of 7.8 × 1014 N+/cm2 at 10 keV, a high-yield PHAs strain with high genetic stability was generated from many mutants. After optimizing its fermentation conditions, the biomass, PHAs concentration and PHAs content of pCBH4 reached 2.26 g/L, 1.81 g/L, and 80.08% respectively, whereas its wild type controls were about 1.24 g/L, 0.61 g/L, and 49.20%. Moreover, the main constituent of PHAs was identified as poly-3-hydroxybutyrates (PHB) in the mutant stain and the yield of this compound was increased up to 41.33% in contrast to that of 27.78% in the wild type strain.

  4. Mapping Crop Yield and Sow Date Using High Resolution Imagery

    NASA Astrophysics Data System (ADS)

    Royal, K.

    2015-12-01

    Keitasha Royal, Meha Jain, Ph.D., David Lobell, Ph.D Mapping Crop Yield and Sow Date Using High Resolution ImageryThe use of satellite imagery in agriculture is becoming increasingly more significant and valuable. Due to the emergence of new satellites, such as Skybox, these satellites provide higher resolution imagery (e.g 1m) therefore improving the ability to map smallholder agriculture. For the smallholder farm dominated area of northern India, Skybox high-resolution satellite imagery can aid in understanding how to improve farm yields. In particular, we are interested in mapping winter wheat in India, as this region produces approximately 80% of the country's wheat crop, which is important given that wheat is a staple crop that provides approximately 20% of household calories. In northeast India, the combination of increased heat stress, limited irrigation access, and the difficulty for farmers to access advanced farming technologies results in farmers only producing about 50% of their potential crop yield. The use of satellite imagery can aid in understanding wheat yields through time and help identify ways to increase crop yields in the wheat belt of India. To translate Skybox satellite data into meaningful information about wheat fields, we examine vegetation indices, such as the normalized difference vegetation index (NDVI), to measure the "greenness" of plants to help determine the health of the crops. We test our ability to predict crop characteristics, like sow date and yield, using vegetation indices of 59 fields for which we have field data in Bihar, India.

  5. Selection of the Mutants with High Hydroquinone Degradation Ability of Serratia Marcesscen by Plasma Mutation

    NASA Astrophysics Data System (ADS)

    Yao, Risheng; You, Qidong; He, Weijing; Zhu, Huixia

    2009-06-01

    In this study, an efficient way by plasma induced mutation was applied to improve the hydroquinone degradation capacity of Serratia marcescens AB 90027 (SM27). The results showed that combined with the selection of hydroquinone tolerance, the mutant with high hydroquinone degradation ability induced by plasma could be achieved. The best dose for plasma mutation was 15 s, which showed a 47.0% higher positive mutation ratio. Besides, the aimed mutant was markedly different from the parent strain (SM27) in colonial traits while cultivated on Kings media. Finally, the hydroquinone degradation ratio reached 70.5% using the induced mutant strain with 1500 mg/L hydroquinone (HQ) after 15 days of cultivation as the selective conditions; however, it was only 46.7% for SM27. The improvement of the degradation capacity by the induced mutant with a high concentration of HQ selection was attributed to its faster growth and higher hydroquinone tolerance compared with that of the parent strain.

  6. Isolation of a highly active photosystem II preparation from Synechocystis 6803 using a histidine-tagged mutant of CP 47.

    PubMed

    Bricker, T M; Morvant, J; Masri, N; Sutton, H M; Frankel, L K

    1998-11-01

    Site-directed mutagenesis was used to produce a Synechocystis mutant containing a histidine tag at the C terminus of the CP 47 protein of Photosystem II. This mutant cell line, designated HT-3, exhibited slightly above normal rates of oxygen evolution and appeared to accumulate somewhat more Photosystem II reaction centers than a control strain. A rapidly isolatable (<7 h) oxygen-evolving Photosystem II preparation was prepared from HT-3 using dodecyl-beta-d-maltoside solubilization and Co2+ metal affinity chromatography. This histidine-tagged Photosystem II preparation stably evolved oxygen at a high rate (2440 micromol O2 (mg chl)-1 h-1), exhibited an alpha-band absorption maximum at 674 nm, and was highly enriched in a number of Photosystem II components including cytochrome c550. Fluorescence yield analysis using water or hydroxylamine as an electron donor to the Photosystem II preparation indicated that virtually all of the Photosystem II reaction centers were capable of evolving oxygen. Proteins associated with Photosystem II were highly enriched in this preparation. 3,3',5, 5'-Tetramethylbenzidine staining indicated that the histidine-tagged preparation was enriched in cytochromes c550 and b559 and depleted of cytochrome f. This result was confirmed by optical difference spectroscopy. This histidine-tagged Photosystem II preparation may be very useful for the isolation of Photosystem II preparations from mutants containing lesions in other Photosystem II proteins. PMID:9804889

  7. Neutron emission and fragment yield in high-energy fission

    SciTech Connect

    Grudzevich, O. T. Klinov, D. A.

    2013-07-15

    The KRIS special library of spectra and emission probabilities in the decays of 1500 nuclei excited up to energies between 150 and 250 MeV was developed for correctly taking into account the decay of highly excited nuclei appearing as fission fragments. The emission of neutrons, protons, and photons was taken into account. Neutron emission fromprimary fragments was found to have a substantial effect on the formation of yields of postneutron nuclei. The library was tested by comparing the calculated and measured yields of products originating from the fission of nuclei that was induced by high-energy protons. The method for calculating these yields was tested on the basis of experimental data on the thermal-neutroninduced fission of {sup 235}U nuclei.

  8. High-yield maize with large net energy yield and small global warming intensity

    PubMed Central

    Grassini, Patricio; Cassman, Kenneth G.

    2012-01-01

    Addressing concerns about future food supply and climate change requires management practices that maximize productivity per unit of arable land while reducing negative environmental impact. On-farm data were evaluated to assess energy balance and greenhouse gas (GHG) emissions of irrigated maize in Nebraska that received large nitrogen (N) fertilizer (183 kg of N⋅ha−1) and irrigation water inputs (272 mm or 2,720 m3 ha−1). Although energy inputs (30 GJ⋅ha−1) were larger than those reported for US maize systems in previous studies, irrigated maize in central Nebraska achieved higher grain and net energy yields (13.2 Mg⋅ha−1 and 159 GJ⋅ha−1, respectively) and lower GHG-emission intensity (231 kg of CO2e⋅Mg−1 of grain). Greater input-use efficiencies, especially for N fertilizer, were responsible for better performance of these irrigated systems, compared with much lower-yielding, mostly rainfed maize systems in previous studies. Large variation in energy inputs and GHG emissions across irrigated fields in the present study resulted from differences in applied irrigation water amount and imbalances between applied N inputs and crop N demand, indicating potential to further improve environmental performance through better management of these inputs. Observed variation in N-use efficiency, at any level of applied N inputs, suggests that an N-balance approach may be more appropriate for estimating soil N2O emissions than the Intergovernmental Panel on Climate Change approach based on a fixed proportion of applied N. Negative correlation between GHG-emission intensity and net energy yield supports the proposition that achieving high yields, large positive energy balance, and low GHG emissions in intensive cropping systems are not conflicting goals. PMID:22232684

  9. High-yield maize with large net energy yield and small global warming intensity.

    PubMed

    Grassini, Patricio; Cassman, Kenneth G

    2012-01-24

    Addressing concerns about future food supply and climate change requires management practices that maximize productivity per unit of arable land while reducing negative environmental impact. On-farm data were evaluated to assess energy balance and greenhouse gas (GHG) emissions of irrigated maize in Nebraska that received large nitrogen (N) fertilizer (183 kg of N · ha(-1)) and irrigation water inputs (272 mm or 2,720 m(3) ha(-1)). Although energy inputs (30 GJ · ha(-1)) were larger than those reported for US maize systems in previous studies, irrigated maize in central Nebraska achieved higher grain and net energy yields (13.2 Mg · ha(-1) and 159 GJ · ha(-1), respectively) and lower GHG-emission intensity (231 kg of CO(2)e · Mg(-1) of grain). Greater input-use efficiencies, especially for N fertilizer, were responsible for better performance of these irrigated systems, compared with much lower-yielding, mostly rainfed maize systems in previous studies. Large variation in energy inputs and GHG emissions across irrigated fields in the present study resulted from differences in applied irrigation water amount and imbalances between applied N inputs and crop N demand, indicating potential to further improve environmental performance through better management of these inputs. Observed variation in N-use efficiency, at any level of applied N inputs, suggests that an N-balance approach may be more appropriate for estimating soil N(2)O emissions than the Intergovernmental Panel on Climate Change approach based on a fixed proportion of applied N. Negative correlation between GHG-emission intensity and net energy yield supports the proposition that achieving high yields, large positive energy balance, and low GHG emissions in intensive cropping systems are not conflicting goals.

  10. Zinc-oxygen primary cell yields high energy density

    NASA Technical Reports Server (NTRS)

    Graff, C. B.

    1968-01-01

    Zinc-oxygen primary cell yields high energy density for battery used as an auxiliary power source in space vehicle systems. Maximum reliability and minimum battery weight is achieved by using a stacking configuration of 23 series-connected modules with 6 parallel-connected cells per module.

  11. Setting the Record Straight on "High-Yield" Strategies

    ERIC Educational Resources Information Center

    Marzano, Robert J.

    2009-01-01

    Widely credited with proposing nine "high-yield" instructional strategies, author Robert J. Marzano sets the record straight about the broader number of strategies identified by the research. He provides a list of 41 strategies and suggests more nuanced ways of using, observing, and evaluating them. (Contains 1 figure.)

  12. Executive Summary High-Yield Scenario Workshop Series Report

    SciTech Connect

    Leslie Park Ovard; Thomas H. Ulrich; David J. Muth Jr.; J. Richard Hess; Steven Thomas; Bryce Stokes

    2009-12-01

    To get a collective sense of the impact of research and development (R&D) on biomass resource availability, and to determine the feasibility that yields higher than baseline assumptions used for past assessments could be achieved to support U.S. energy independence, an alternate “High-Yield Scenario” (HYS) concept was presented to industry experts at a series of workshops held in December 2009. The workshops explored future production of corn/agricultural crop residues, herbaceous energy crops (HECs), and woody energy crops (WECs). This executive summary reports the findings of that workshop.

  13. TIO2 Based Electrorheological Fluid with High Yield Stress

    NASA Astrophysics Data System (ADS)

    Shen, Rong; Wang, Xuezhao; Wen, Weijia; Lu, Kunquan

    We have fabricated several TiO2 based ER fluids with doping and without designed doping, which exhibit the high yield stress up to more than 100kPa. The titanium oxide nanoparticles were synthesized by using wet chemical method. The ER effect of those materials is dominated by the special additives, such as amide or its ramification, as well as the remained molecules or ions in the sample preparation. It is found that the yield stress is also strongly dependent on the viscosity of the oil. The prepared ER fluids possess other attractive characters, for instance the current density is low and against sedimentation.

  14. Development of DNA markers for newly identified high-oleate peanut mutants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Development of high-oleate cultivars is one of the important objectives of peanut breeding because consuming products containing high oleate can benefit human health in many aspects. By screening the entire USDA cultivated peanut collection, we have identified two new high-oleate mutants (PI 342664...

  15. Enhancing the growth rate and astaxanthin yield of Haematococcus pluvialis by nuclear irradiation and high concentration of carbon dioxide stress.

    PubMed

    Cheng, Jun; Li, Ke; Yang, Zongbo; Zhou, Junhu; Cen, Kefa

    2016-03-01

    Unicellular green microalgae Haematococcus pluvialis was mutated with (60)Co-γ irradiation to promote growth rate and increase astaxanthin yield under high concentration of CO2 stress. The average specific growth rate of H. pluvialis mutated with 4000 Gy γ-ray irradiation was increased by 15% compared with the original strain with air aeration. The mutant grew best with 6% CO2 (the maximum specific growth rate was 0.60/d) when it was cultured with high concentrations of CO2 (2-10%). The peak biomass productivity (0.16 g/L/d) of the mutant cultured with 6% CO2 was 82% higher than that of the mutant with air. The astaxanthin yield and lipid content of the mutant induced with 6% CO2 and high light (108 μmol photons m(-2) s(-1)) increased to 46.0mg/L and 45.9%, which were 2.4 and 1.3 times higher than those of the wild-type strain, respectively.

  16. Ascorbate-deficient mutants of Arabidopsis grow in high light despite chronic photooxidative stress.

    PubMed

    Müller-Moulé, Patricia; Golan, Talila; Niyogi, Krishna K

    2004-03-01

    Acclimation to changing environments, such as increases in light intensity, is necessary, especially for the survival of sedentary organisms like plants. To learn more about the importance of ascorbate in the acclimation of plants to high light (HL), vtc2, an ascorbate-deficient mutant of Arabidopsis, and the double mutants vtc2npq4 and vtc2npq1 were tested for growth in low light and HL and compared with the wild type. The vtc2 mutant has only 10% to 30% of wild-type levels of ascorbate, vtc2npq4 has lower ascorbate levels and lacks non-photochemical quenching of chlorophyll fluorescence (NPQ) because of the absence of the photosystem II protein PsbS, and vtc2npq1 is NPQ deficient and also lacks zeaxanthin in HL but has PsbS. All three genotypes were able to grow in HL and had wild-type levels of Lhcb1, cytochrome f, PsaF, and 2-cysteine peroxiredoxin. However, the mutants had lower electron transport and oxygen evolution rates and lower quantum efficiency of PSII compared with the wild type, implying that they experienced chronic photooxidative stress. The mutants lacking NPQ in addition to ascorbate were only slightly more affected than vtc2. All three mutants had higher glutathione levels than the wild type in HL, suggesting a possible compensation for the lower ascorbate content. These results demonstrate the importance of ascorbate for the long-term acclimation of plants to HL.

  17. Formation of oligopeptides in high yield under simple programmable conditions

    PubMed Central

    Rodriguez-Garcia, Marc; Surman, Andrew J.; Cooper, Geoffrey J.T.; Suárez-Marina, Irene; Hosni, Zied; Lee, Michael P.; Cronin, Leroy

    2015-01-01

    Many high-yielding reactions for forming peptide bonds have been developed but these are complex, requiring activated amino-acid precursors and heterogeneous supports. Herein we demonstrate the programmable one-pot dehydration–hydration condensation of amino acids forming oligopeptide chains in around 50% yield. A digital recursive reactor system was developed to investigate this process, performing these reactions with control over parameters such as temperature, number of cycles, cycle duration, initial monomer concentration and initial pH. Glycine oligopeptides up to 20 amino acids long were formed with very high monomer-to-oligomer conversion, and the majority of these products comprised three amino acid residues or more. Having established the formation of glycine homo-oligopeptides, we then demonstrated the co-condensation of glycine with eight other amino acids (Ala, Asp, Glu, His, Lys, Pro, Thr and Val), incorporating a range of side-chain functionality. PMID:26442968

  18. High-throughput phenotyping of chlamydomonas swimming mutants based on nanoscale video analysis.

    PubMed

    Fujita, Shohei; Matsuo, Takuya; Ishiura, Masahiro; Kikkawa, Masahide

    2014-07-15

    Studies on biflagellated algae Chlamydomonas reinhardtii mutants have resulted in significant contributions to our understanding of the functions of cilia/flagella components. However, visual inspection conducted under a microscope to screen and classify Chlamydomonas swimming requires considerable time, effort, and experience. In addition, it is likely that identification of mutants by this screening is biased toward individual cells with severe swimming defects, and mutants that swim slightly more slowly than wild-type cells may be missed by these screening methods. To systematically screen Chlamydomonas swimming mutants, we have here developed the cell-locating-with-nanoscale-accuracy (CLONA) method to identify the cell position to within 10-nm precision through the analysis of high-speed video images. Instead of analyzing the shape of the flagella, which is not always visible in images, we determine the position of Chlamydomonas cell bodies by determining the cross-correlation between a reference image and the image of the cell. From these positions, various parameters related to swimming, such as velocity and beat frequency, can be accurately estimated for each beat cycle. In the examination of wild-type and seven dynein arm mutants of Chlamydomonas, we found characteristic clustering on scatter plots of beat frequency versus swimming velocity. Using the CLONA method, we have screened 38 Chlamydomonas strains and detected believed-novel motility-deficient mutants that would be missed by visual screening. This CLONA method can automate the screening for mutants of Chlamydomonas and contribute to the elucidation of the functions of motility-associated proteins.

  19. High-Yield Magnetized Liner Fusion Explosions and Blast Mitigation

    NASA Astrophysics Data System (ADS)

    Slutz, Stephen; Vesey, Roger; Cuneo, Michael

    2011-10-01

    Cylindrical liner implosions with preheated and magnetized deuterium-tritium (DT) are predicted to reach fusion conditions on present pulsed power machines [S.A. Slutz et al Phys. Plasmas 17, 056303 (2010)]. We present simulations indicating that high yields (1-10 GJ) and gains (100-1000) may be possible at currents of about 60-70 MA if a cryogenic layer of solid DT is provided on the inside surface of the metal liner. A hot spot is formed from the central preheated magnetized low-density gas and a burn wave propagates radially into the surrounding cold dense fuel. These yields and gains are more than adequate for inertial fusion energy. However, the pulsed-power driver must be protected from the blast of these high-yield explosions. Numerical simulations are presented which show that the blast can be deflected and the fusion neutrons absorbed by a blanket that partially surrounds the liner. Thus a modest length transmission line can be used to deliver power to the liner. Sandia is a multiprogram laboratory operated by Sandia Corporation, a Lockheed Martin Company, for the United States Department of Energy's National Nuclear Security Administration under contract DE-AC04-94AL85000.

  20. Laser heating challenges of high yield MagLIF targets

    NASA Astrophysics Data System (ADS)

    Slutz, Stephen; Sefkow, Adam; Vesey, Roger

    2014-10-01

    The MagLIF (Magnetized Liner Inertial Fusion) concept is predicted by numerical simulation to produce fusion yields of about 100 kJ, when driven by 25 MA from the existing Z accelerator [S. A. Slutz et al. Phys. Plasmas 17, 056303 (2010)] and much higher yields with future accelerators delivering higher currents [Slutz and Vesey PRL 108, 025003 (2012)]. The fuel must be heated before compression to obtain significant fusion yields due to the relatively slow implosion velocities (~ 100 km/s) of magnetically driven liners. Lasers provide a convenient means to accomplish this pre-compressional heating of the fusion fuel, but there are challenges. The laser must penetrate a foil covering the laser entrance hole and deposit 20-30 kJ within the ~1 cm length of the liner in fuel at 6-12 mg/cc. Such high densities could result in beam scattering due to refraction and laser plasma interactions. Numerical simulations of the laser heating process are presented, which indicate that energies as high as 30 kJ could be deposited in the fuel by using two laser pulses of different wavelengths. Simulations of this process will be presented as well of results for a MagLIF design for a potential new machine delivering 50 MA of current. Sandia is a multiprogram laboratory operated by Sandia Corporation, a Lockheed Martin Company, for the United States Department of Energy's National Nuclear Security Administration under Contract DE-AC04-94AL85000.

  1. [Isolation of a high hydrogen-producing mutant TB34 generated by transposon insertion and analysis of hydrogen production].

    PubMed

    Liu, Hong-Yan; Wang, Guang-Ce; Shi, Liu-Yang; Zhu, Da-Ling

    2012-07-01

    To increase the hydrogen-producing capacity of Pantoea agglomerans BH18, isolated from mangrove sludge, we constructed a stable transposon mutagenesis library of this strain. A Tn7-based transposon was randomly inserted into the genomic DNA. Mutants were screened by kanamycin resistance and identified by amplification of the inserted transposon sequences. A mutant strain TB34 was isolated, whose hydrogen production capacity was significantly improved compared to the wild type strain. In seawater-containing medium supplemented with 10 g x L(-1) glucose and had an initial pH of 7.0, the hydrogen yield (H2/glucose) of the mutant strain was (2.04 +/- 0.04) mol x mol(-1), which was 43% higher than that of the wild type strain. The mutant TB34 showed steady hydrogen production capacity for five consecutive passages. Different carbon sources were tested in the hydrogen production by the mutant TB34 and the results showed that both the mutant strain TB34 and the wild type strain BH18 were able to produce hydrogen on sucrose, glucose and fructose. However, different from the wild type strain, the mutant strain TB34 was also able to produce hydrogen using xylose as substrate, with a hydrogen yield (H2/xylose) of (1.34 +/- 0.09) mol x mol(-1), indicating a broader substrate spectrum in the mutant.

  2. An alternate high yielding purification method for Clitoria ternatea lectin.

    PubMed

    Naeem, Aabgeena; Ahmad, Ejaz; Khan, Rizwan Hasan

    2007-10-01

    In our previous publication we had reported the purification and characterization of Clitoria ternatea agglutinin from its seeds on fetuin CL agarose affinity column, designated CTA [A. Naeem, S. Haque, R.H. Khan. Protein J., 2007]. Since CTA binds beta-d-galactosides, this lectin can be used as valuable tool for glycobiology studies in biomedical and cancer research. So an attempt was made for a high yielding alternative purification method employing the use of asialofetuin CL agarose column for the above-mentioned lectin, designated CTL. The fetuin affinity purified agglutinin was found similar to asialofetuin affinity purified lectin in SDS pattern, HPLC and N-terminal sequence. The content of lectin was found to be 30mg/30g dry weight of pulse. The yield was 2.8% as compared to 0.3% obtained on fetuin column. The number of tryptophan and tyrosine estimated was four and six per subunit. PMID:17590430

  3. The role of the high potential form of the cytochrome b559: Study of Thermosynechococcus elongatus mutants.

    PubMed

    Guerrero, Fernando; Zurita, Jorge L; Roncel, Mercedes; Kirilovsky, Diana; Ortega, José M

    2014-06-01

    Cytochrome b559 is an essential component of the photosystem II reaction center in photosynthetic oxygen-evolving organisms, but its function still remains unclear. The use of photosystem II preparations from Thermosynechococcus elongatus of high integrity and activity allowed us to measure for the first time the influence of cytochrome b559 mutations on its midpoint redox potential and on the reduction of the cytochrome b559 by the plastoquinone pool (or QB). In this work, five mutants having a mutation in the α-subunit (I14A, I14S, R18S, I27A and I27T) and one in the β-subunit (F32Y) of cytochrome b559 have been investigated. All the mutations led to a destabilization of the high potential form of the cytochrome b559. The midpoint redox potential of the high potential form was significantly altered in the αR18S and αI27T mutant strains. The αR18S strain also showed a high sensitivity to photoinhibitory illumination and an altered oxidase activity. This was suggested by measurements of light induced oxidation and dark re-reduction of the cytochrome b559 showing that under conditions of a non-functional water oxidation system, once the cytochrome is oxidized by P680(+), the yield of its reduction by QB or the PQ pool was smaller and the kinetic slower in the αR18S mutant than in the wild-type strain. Thus, the extremely positive redox potential of the high potential form of cytochrome b559 could be necessary to ensure efficient oxidation of the PQ pool and to function as an electron reservoir replacing the water oxidation system when it is not operating.

  4. Chimeric neuraminidase and mutant PB1 gene constellation improves growth and yield of H5N1 vaccine candidate virus.

    PubMed

    Plant, Ewan P; Ye, Zhiping

    2015-04-01

    We previously showed that a mutated PB1 gene improved the growth kinetics of a H3N2 influenza reassortant. Here, we showed that the same mutations improved the growth kinetics of a virus containing the A/Vietnam/1203/2004 (H5N1) haemagglutinin and neuraminidase (NA). Total protein yield and NA activity were increased when a chimeric NA was included. These increases indicated that the synergistic effect was due to the gene constellation containing both the altered PB1 gene and the chimeric NA gene.

  5. Adaptation of Chlamydomonas reinhardtii high-CO sub 2 -requiring mutants to limiting CO sub 2

    SciTech Connect

    Suzuki, K.; Spalding, M.H. )

    1989-07-01

    Photosynthetic characteristics of four high-CO{sub 2}-requiring mutants of Chlamydomonas reinhardtii were compared to those of wild type before and after a 24-hour exposure to limiting CO{sub 2} concentrations. The four mutants represent two loci involved in the CO{sub 2}-concentrating system of this unicellular alga. All mutants had a lower photosynthetic affinity for inorganic carbon than did the wild type when grown at an elevated CO{sub 2} concentration, indicating that the genetic lesion in each is expressed even at elevated CO{sub 2} concentrations. Wild type and all four mutants exhibited adaptive responses to limiting CO{sub 2} characteristic of the induction of the CO{sub 2}-concentrating system, resulting in an increased affinity for inorganic carbon only in wild type. Although other components of the CO{sub 2}-concentrating system were induced in these mutants, the defective component in each was sufficient to prevent any increase in the affinity for inorganic carbon. It was concluded that the genes corresponding to the ca-1 and pmp-1 loci exhibit at least partially constitutive expression and that all components of the CO{sub 2}-concentrating system may be required to significantly affect the photosynthetic affinity for inorganic carbon.

  6. Increase in ethanol yield via elimination of lactate production in an ethanol-tolerant mutant of Clostridium thermocellum

    SciTech Connect

    Biswas, Ranjita; Prabhu, Sandeep; Lynd, Lee R; Guss, Adam M

    2014-01-01

    Large-scale production of lignocellulosic biofuel is a potential solution to sustainably meet global energy needs. One-step consolidated bioprocessing (CBP) is a potentially advantageous approach for the production of biofuels, but requires an organism capable of hydrolyzing biomass to sugars and fermenting the sugars to ethanol at commercially viable titers and yields. Clostridium thermocellum, a thermophilic anaerobe, can ferment cellulosic biomass to ethanol and organic acids, but low yield, low titer, and ethanol sensitivity remain barriers to industrial production. Here, we deleted the hypoxanthine phosphoribosyltransferase gene in ethanol tolerant strain of C. thermocellum adhE*(EA) in order to allow use of previously developed gene deletion tools, then deleted lactate dehydrogenase (ldh) to redirect carbon flux towards ethanol. Upon deletion of ldh, the adhE*(EA) ldh strain produced 30% more ethanol than wild type on minimal medium. The adhE*(EA) ldh strain retained tolerance to 5% v/v ethanol, resulting in an ethanol tolerant platform strain of C. thermocellum for future metabolic engineering efforts.

  7. Isolation and characterization of high-temperature-induced Dauer formation mutants in Caenorhabditis elegans.

    PubMed Central

    Ailion, Michael; Thomas, James H

    2003-01-01

    Dauer formation in Caenorhabditis elegans is regulated by at least three signaling pathways, including an insulin receptor-signaling pathway. These pathways were defined by mutants that form dauers constitutively (Daf-c) at 25 degrees. Screens for Daf-c mutants at 25 degrees have probably been saturated, but failed to identify all the components involved in regulating dauer formation. Here we screen for Daf-c mutants at 27 degrees, a more strongly dauer-inducing condition. Mutations identified include novel classes of alleles for three known genes and alleles defining at least seven new genes, hid-1-hid-7. Many of the genes appear to act in the insulin branch of the dauer pathway, including pdk-1, akt-1, aex-6, and hid-1. We also molecularly identify hid-1 and show that it encodes a novel highly conserved putative transmembrane protein expressed in neurons. PMID:14504222

  8. Projecting crop yield in northern high latitude area.

    PubMed

    Matsumura, Kanichiro

    2014-01-01

    Changing climatic conditions on seasonal and longer time scales influence agricultural production. Improvement of soil and fertilizer is a strong factor in agricultural production, but agricultural production is influenced by climate conditions even in highly developed countries. It is valuable if fewer predictors make it possible to conduct future projections. Monthly temperature and precipitation, wintertime 500hPa geopotential height, and the previous year's yield are used as predictors to forecast spring wheat yield in advance. Canadian small agricultural divisions (SAD) are used for analysis. Each SAD is composed of a collection of Canadian Agricultural Regions (CAR) of similar weather and growing conditions. Spring wheat yields in each CAR are forecast from the following variables: (a) the previous year's yield, (b) earlier stages of the growing season's climate conditions and, (c) the previous year's wintertime northern hemisphere 500hPa geopotential height field. Arctic outflow events in the Okanagan Valley in Canada are associated with episodes of extremely low temperatures during wintertime. Principal component analysis (PCA) is applied for wintertime northern hemisphere 500hPa geopotential height anomalies. The spatial PCA mode1 is defined as Arctic Oscillation and it influences prevailing westerlies. The prevailing westerlies meanders and influences climatic conditions. The spatial similarity between wintertime top 5 Arctic outflow event year's composites of 500hPa geopotential height anomalies and mode 3's spatial pattern is found. Mode 3's spatial pattern looks like the Pacific/North American (PNA) pattern which describes the variation of atmospheric circulation pattern over the Pacific Ocean and North America. Climate conditions from April to June, May to July, mode 3's time coefficients, and previous year's yield are used for forecasting spring wheat yield in each SAD. Cross-validation procedure which generates eight sets of models for the eight

  9. A high-yield saponification of galactosylceramide I(3)-sulfate.

    PubMed

    Koshy, K M; Boggs, J M

    1982-12-01

    A method for the deacylation of galactosylceramide I(3)-sulphate using aqueous methanolic KOH is described. The combination of a relatively low concentration of the alkali (0.3 M) and a moderate reaction temperature (reflux point of 90% methanol) results in the formation of galactosylsphingosine I(3)-sulphate in consistently high yields (61%) with a minimum of side reactions. The product was purified by column chromatography and its identity established by thin layer chromatography, infrared spectroscopy, determination of galactose content and organic sulphate assay using established methods or their modifications.

  10. Functional and fluorochrome analysis of an exocytotic mutant yields evidence of store-operated Ca2+ influx in Paramecium.

    PubMed

    Mohamed, I; Klauke, N; Hentschel, J; Cohen, J; Plattner, H

    2002-05-01

    A non-discharge mutant of Paramecium tetraurelia (nd12-35 degrees C, lacking exocytotic response upon stimulation with the nonpermeable polycationic secretagogue aminoethyldextran, AED), in the pawnA genetic context (d4-500r, lacking ciliary voltage-dependent Ca2+ influx), was shown to lack (45)Ca2+ entry from outside upon AED stimulation. In contrast, cells grown at 25 degrees C behave like the wildtype. To check the functional properties in more detail, fluorochrome-loaded 35 degrees C cells were stimulated, not only with AED (EC(100) = 10(-6) M in wildtype cells), but also with 4-chloro-meta-cresol, (4CmC, 0.5 mM), a permeable activator of ryanodine receptor-type Ca2+ release channels, usually at extracellular [Ca2+] of 50 microM, and eventually with a Ca2+ chelator added. We confirm that pwA-nd12(35 degrees C) cells lack any Ca2+ influx and any exocytosis of trichocysts in response to any stimulus. As we determined by x-ray microanalysis, total calcium content in alveolar sacs (subplasmalemmal stores) known to be mobilized upon exocytosis stimulation in wild-type cells, contain about the same total calcium in 35 degrees C as in 25 degrees C cells, and Ca2+ mobilization from alveoli by AED or 4CmC is also nearly the same. Due to the absence of any AED-induced Ca2+ influx in 35 degrees C cells and normal Ca2+ release from stores found by x-ray microanalysis one can exclude a "CICR"-type mechanism (Ca2+-induced Ca2+ release) and imply that normally a store-operated Ca2+ ("SOC") influx would occur (as in 25 degrees C cells). Furthermore, 35 degrees C cells display a significantly lower basal intracellular [Ca2+], so that any increase upon stimulation may be less expressed or even remain undetected. Under these conditions, any mobilization of Ca2+ from stores cannot compensate for the lack of Ca2+ influx, particularly since normally both components have to cooperate to achieve full exocytotic response. Also striking is our finding that 35 degrees C cells are unable

  11. Designing a mutant CCL2-HSA chimera with high glycosaminoglycan-binding affinity and selectivity.

    PubMed

    Gerlza, Tanja; Winkler, Sophie; Atlic, Aid; Zankl, Christina; Konya, Viktoria; Kitic, Nikola; Strutzmann, Elisabeth; Knebl, Kerstin; Adage, Tiziana; Heinemann, Akos; Weis, Roland; Kungl, Andreas J

    2015-08-01

    Chemokines like CCL2 mediate leukocyte migration to inflammatory sites by binding to G-protein coupled receptors on the target cell as well as to glycosaminoglycans (GAGs) on the endothelium of the inflamed tissue. We have recently shown that the dominant-negative Met-CCL2 mutant Y13A/S21K/Q23R with improved GAG binding affinity is highly bio-active in several animal models of inflammatory diseases. For chronic indications, we have performed here a fusion to human serum albumin (HSA) in order to extend the serum half-life of the chemokine mutant. To compensate a potential drop in GAG-binding affinity due to steric hindrance by HSA, a series of novel CCL2 mutants was generated with additional basic amino acids which were genetically introduced at sites oriented towards the GAG ligand. From this set of mutants, the Met-CCL2 variant Y13A/N17K/S21K/Q23K/S34K exhibited high GAG-binding affinity and a similar selectivity as wild type (wt) CCL2. From a set of different HSA-chemokine chimeric constructs, the linked HSA(C34A)(Gly)4Ser-Met-CCL2(Y13A/N17K/S21K/Q23K/S34K) fusion protein was found to show the best overall GAG-binding characteristics. Molecular modeling demonstrated an energetically beneficial fold of this novel protein chimera. This was experimentally supported by GdmCl-induced unfolding studies, in which the fusion construct exhibited a well-defined secondary structure and a transition point significantly higher than both the wt and the unfused CCL2 mutant protein. Unlike the wt chemokine, the quaternary structure of the HSA-fusion protein is monomeric according to size-exclusion chromatography experiments. In competition experiments, the HSA-fusion construct displaced only two of seven unrelated chemokines from heparan sulfate, whereas the unfused CCL2 mutant protein displaced five other chemokines. The most effective concentration of the HSA-fusion protein in inhibiting CCL2-mediated monocyte attachment to endothelial cells, as detected in the flow chamber

  12. Development of a mutant strain of Escherichia coli for molecular cloning of highly methylated DNA

    SciTech Connect

    Bishr, M.A.

    1991-01-01

    A mutant strain of Escherichia coli designated as GR219 that allows efficient molecular cloning of highly methylated bean DNA has been developed by UV light mutation of the parent LE392 str[sup r] strain. This mutant strain, like the parent, is streptomycin resistant and is biologically contained, because it requires thymidine for growth. Both the wild type and the mutant strain have lambda phage receptors so both can be utilized for construction of genomic libraries using the phase as a vector. The efficiency of transformation of the parent and the mutant strain with a recombinant plasmid containing bean DNA was compared to the efficiency of transformation of the PLK-F[prime] strain, which has a deletion of mcrA and mcrB genes and, therefore, allows transformation with methylated bean DNA. It has been found that the GR219 strain has the highest efficiency of transformation, while the PLK-F[prime] strain shows less, and the parent LE392 str[sup r] strain the least efficiency of transformation. These results indicate that strains of E. coli with mcrA and mcrB genes can recognize and degrade highly methylated DNA. However, other undefined factors affected by the altered gene(s) in the GR219 strain are also involved in the recognition and degradation of any cloned foreign DNA.

  13. A fluorescence-activated cell sorting-based strategy for rapid isolation of high-lipid Chlamydomonas mutants.

    PubMed

    Terashima, Mia; Freeman, Elizabeth S; Jinkerson, Robert E; Jonikas, Martin C

    2015-01-01

    There is significant interest in farming algae for the direct production of biofuels and valuable lipids. Chlamydomonas reinhardtii is the leading model system for studying lipid metabolism in green algae, but current methods for isolating mutants of this organism with a perturbed lipid content are slow and tedious. Here, we present the Chlamydomonas high-lipid sorting (CHiLiS) strategy, which enables enrichment of high-lipid mutants by fluorescence-activated cell sorting (FACS) of pooled mutants stained with the lipid-sensitive dye Nile Red. This method only takes 5 weeks from mutagenesis to mutant isolation. We developed a staining protocol that allows quantification of lipid content while preserving cell viability. We improved separation of high-lipid mutants from the wild type by using each cell's chlorophyll fluorescence as an internal control. We initially demonstrated 20-fold enrichment of the known high-lipid mutant sta1 from a mixture of sta1 and wild-type cells. We then applied CHiLiS to sort thousands of high-lipid cells from a pool of about 60,000 mutants. Flow cytometry analysis of 24 individual mutants isolated by this approach revealed that about 50% showed a reproducible high-lipid phenotype. We further characterized nine of the mutants with the highest lipid content by flame ionization detection and mass spectrometry lipidomics. All mutants analyzed had a higher triacylglycerol content and perturbed whole-cell fatty acid composition. One arbitrarily chosen mutant was evaluated by microscopy, revealing larger lipid droplets than the wild type. The unprecedented throughput of CHiLiS opens the door to a systems-level understanding of green algal lipid biology by enabling genome-saturating isolation of mutants in key genes.

  14. A fluorescence-activated cell sorting-based strategy for rapid isolation of high-lipid Chlamydomonas mutants

    PubMed Central

    Terashima, Mia; Freeman, Elizabeth S; Jinkerson, Robert E; Jonikas, Martin C

    2015-01-01

    There is significant interest in farming algae for the direct production of biofuels and valuable lipids. Chlamydomonas reinhardtii is the leading model system for studying lipid metabolism in green algae, but current methods for isolating mutants of this organism with a perturbed lipid content are slow and tedious. Here, we present the Chlamydomonas high-lipid sorting (CHiLiS) strategy, which enables enrichment of high-lipid mutants by fluorescence-activated cell sorting (FACS) of pooled mutants stained with the lipid-sensitive dye Nile Red. This method only takes 5 weeks from mutagenesis to mutant isolation. We developed a staining protocol that allows quantification of lipid content while preserving cell viability. We improved separation of high-lipid mutants from the wild type by using each cell's chlorophyll fluorescence as an internal control. We initially demonstrated 20-fold enrichment of the known high-lipid mutant sta1 from a mixture of sta1 and wild-type cells. We then applied CHiLiS to sort thousands of high-lipid cells from a pool of about 60 000 mutants. Flow cytometry analysis of 24 individual mutants isolated by this approach revealed that about 50% showed a reproducible high-lipid phenotype. We further characterized nine of the mutants with the highest lipid content by flame ionization detection and mass spectrometry lipidomics. All mutants analyzed had a higher triacylglycerol content and perturbed whole-cell fatty acid composition. One arbitrarily chosen mutant was evaluated by microscopy, revealing larger lipid droplets than the wild type. The unprecedented throughput of CHiLiS opens the door to a systems-level understanding of green algal lipid biology by enabling genome-saturating isolation of mutants in key genes. PMID:25267488

  15. A fluorescence-activated cell sorting-based strategy for rapid isolation of high-lipid Chlamydomonas mutants.

    PubMed

    Terashima, Mia; Freeman, Elizabeth S; Jinkerson, Robert E; Jonikas, Martin C

    2015-01-01

    There is significant interest in farming algae for the direct production of biofuels and valuable lipids. Chlamydomonas reinhardtii is the leading model system for studying lipid metabolism in green algae, but current methods for isolating mutants of this organism with a perturbed lipid content are slow and tedious. Here, we present the Chlamydomonas high-lipid sorting (CHiLiS) strategy, which enables enrichment of high-lipid mutants by fluorescence-activated cell sorting (FACS) of pooled mutants stained with the lipid-sensitive dye Nile Red. This method only takes 5 weeks from mutagenesis to mutant isolation. We developed a staining protocol that allows quantification of lipid content while preserving cell viability. We improved separation of high-lipid mutants from the wild type by using each cell's chlorophyll fluorescence as an internal control. We initially demonstrated 20-fold enrichment of the known high-lipid mutant sta1 from a mixture of sta1 and wild-type cells. We then applied CHiLiS to sort thousands of high-lipid cells from a pool of about 60,000 mutants. Flow cytometry analysis of 24 individual mutants isolated by this approach revealed that about 50% showed a reproducible high-lipid phenotype. We further characterized nine of the mutants with the highest lipid content by flame ionization detection and mass spectrometry lipidomics. All mutants analyzed had a higher triacylglycerol content and perturbed whole-cell fatty acid composition. One arbitrarily chosen mutant was evaluated by microscopy, revealing larger lipid droplets than the wild type. The unprecedented throughput of CHiLiS opens the door to a systems-level understanding of green algal lipid biology by enabling genome-saturating isolation of mutants in key genes. PMID:25267488

  16. High-Yield Secretion of Multiple Client Proteins in Aspergillus

    SciTech Connect

    Segato, F.; Damasio, A. R. L.; Goncalves, T. A.; de Lucas, R. C.; Squina, F. M.; Decker, S. R.; Prade, R. A.

    2012-07-15

    Production of pure and high-yield client proteins is an important technology that addresses the need for industrial applications of enzymes as well as scientific experiments in protein chemistry and crystallization. Fungi are utilized in industrial protein production because of their ability to secrete large quantities of proteins. In this study, we engineered a high-expression-secretion vector, pEXPYR that directs proteins towards the extracellular medium in two Aspergillii host strains, examine the effect of maltose-induced over-expression and protein secretion as well as time and pH-dependent protein stability in the medium. We describe five client proteins representing a core set of hemicellulose degrading enzymes that accumulated up to 50-100 mg/L of protein. Using a recyclable genetic marker that allows serial insertion of multiple genes, simultaneous hyper-secretion of three client proteins in a single host strain was accomplished.

  17. High-Yield D-T Neutron Generator

    SciTech Connect

    Ludewigt, B.A.; Wells, R.P.; Reijonen, J.

    2006-11-15

    A high-yield D-T neutron generator has been developed for neutron interrogation in homeland security applications such as cargo screening. The generator has been designed as a sealed tube with a performance goal of producing 5 {center_dot} 10{sup 11} n/s over a long lifetime. The key generator components developed are a radio-frequency (RF) driven ion source and a beam-loaded neutron production target that can handle a beam power of 10 kW. The ion source can provide a 100 mA D{sup +}/T{sup +} beam current with a high fraction of atomic species and can be pulsed up to frequencies of several kHz for pulsed neutron generator operation. Testing in D-D operation has been started.

  18. Processable high-carbon-yielding polymer for micro- and nanofabrication

    NASA Astrophysics Data System (ADS)

    Perpall, Mark W.; Zengin, Huseyin; Perera, K. Prasanna U.; Zhou, Wensheng; Shah, Hiren; Wu, Xinyu; Creager, Stephen E.; Smith, Dennis W., Jr.; Foulger, Stephen H.; Ballato, John M.

    2003-01-01

    Bis-ortho-Diynyl Arene (BODA) monomers polymerize to network polynapthalene by the thermally-driven Bergman cyclization and subsequent radical polymerization via oligomeric intermediates that can be melt or solution processed. Further heating of the network to 1000 °C affords a high-yield glassy carbon structure that retains the approximate size and dimensions of the polymer precursor. The higher carbon-yield for BODA networks (75- 80 % by mass) is significantly greater than that of traditional phenol-formaldehyde resins and other carbon precursor polymers leading to its greater dimensional stability. Phenyl terminated BODA derived polymers were fabricated using microprocessing such as the micromolding in capillaries (MIMIC) technique, direct microtransfer molding, and molding in quartz capillary tubes. Nano-scale fabrication using closed packed silica spheres as templates was demonstrated with an hydroxy-terminated monomer which exhibits greatly enhanced compatibility for silica surfaces. After pyrolysis to glassy carbon, the silica is chemically etched leaving an inverse carbon opal photonic crystal which is electrically conductive. The wavelength of light diffracted is a function of the average refractive index of the carbon/ filler composite, which can be modified for use as sensitive detector elements.

  19. Multiple resistance mechanisms among Aspergillus fumigatus mutants with high-level resistance to itraconazole.

    PubMed

    Nascimento, Adriana M; Goldman, Gustavo H; Park, Steven; Marras, Salvatore A E; Delmas, Guillaume; Oza, Uma; Lolans, Karen; Dudley, Michael N; Mann, Paul A; Perlin, David S

    2003-05-01

    A collection of Aspergillus fumigatus mutants highly resistant to itraconazole (RIT) at 100 micro g ml(-1) were selected in vitro (following UV irradiation as a preliminary step) to investigate mechanisms of drug resistance in this clinically important pathogen. Eight of the RIT mutants were found to have a mutation at Gly54 (G54E, -K, or -R) in the azole target gene CYP51A. Primers designed for highly conserved regions of multidrug resistance (MDR) pumps were used in reverse transcriptase PCR amplification reactions to identify novel genes encoding potential MDR efflux pumps in A. fumigatus. Two genes, AfuMDR3 and AfuMDR4, showed prominent changes in expression levels in many RIT mutants and were characterized in more detail. Analysis of the deduced amino acid sequence encoded by AfuMDR3 revealed high similarity to major facilitator superfamily transporters, while AfuMDR4 was a typical member of the ATP-binding cassette superfamily. Real-time quantitative PCR with molecular beacon probes was used to assess expression levels of AfuMDR3 and AfuMDR4. Most RIT mutants showed either constitutive high-level expression of both genes or induction of expression upon exposure to itraconazole. Our results suggest that overexpression of one or both of these newly identified drug efflux pump genes of A. fumigatus and/or selection of drug target site mutations are linked to high-level itraconazole resistance and are mechanistic considerations for the emergence of clinical resistance to itraconazole.

  20. Film quantum yields of EUV& ultra-high PAG photoresists

    SciTech Connect

    Hassanein, Elsayed; Higgins, Craig; Naulleau, Patrick; Matyi, Richard; Gallatin, Greg; Denbeaux, Gregory; Antohe, Alin; Thackery, Jim; Spear, Kathleen; Szmanda, Charles; Anderson, Christopher N.; Niakoula, Dimitra; Malloy, Matthew; Khurshid, Anwar; Montgomery, Cecilia; Piscani, Emil C.; Rudack, Andrew; Byers, Jeff; Ma, Andy; Dean, Kim; Brainard, Robert

    2008-01-10

    Base titration methods are used to determine C-parameters for three industrial EUV photoresist platforms (EUV-2D, MET-2D, XP5496) and twenty academic EUV photoresist platforms. X-ray reflectometry is used to measure the density of these resists, and leads to the determination of absorbance and film quantum yields (FQY). Ultrahigh levels ofPAG show divergent mechanisms for production of photo acids beyond PAG concentrations of 0.35 moles/liter. The FQY of sulfonium PAGs level off, whereas resists prepared with iodonium PAG show FQY s that increase beyond PAG concentrations of 0.35 moles/liter, reaching record highs of 8-13 acids generatedlEUV photons absorbed.

  1. High relatedness maintains multicellular cooperation in a social amoeba by controlling cheater mutants

    PubMed Central

    Gilbert, Owen M.; Foster, Kevin R.; Mehdiabadi, Natasha J.; Strassmann, Joan E.; Queller, David C.

    2007-01-01

    The control of cheating is important for understanding major transitions in evolution, from the simplest genes to the most complex societies. Cooperative systems can be ruined if cheaters that lower group productivity are able to spread. Kin-selection theory predicts that high genetic relatedness can limit cheating, because separation of cheaters and cooperators limits opportunities to cheat and promotes selection against low-fitness groups of cheaters. Here, we confirm this prediction for the social amoeba Dictyostelium discoideum; relatedness in natural wild groups is so high that socially destructive cheaters should not spread. We illustrate in the laboratory how high relatedness can control a mutant that would destroy cooperation at low relatedness. Finally, we demonstrate that, as predicted, mutant cheaters do not normally harm cooperation in a natural population. Our findings show how altruism is preserved from the disruptive effects of such mutant cheaters and how exceptionally high relatedness among cells is important in promoting the cooperation that underlies multicellular development. PMID:17496139

  2. Production of Highly Sialylated Recombinant Glycoproteins Using Ricinus communis Agglutinin-I-Resistant CHO Glycosylation Mutants.

    PubMed

    Goh, John S Y; Chan, Kah Fai; Song, Zhiwei

    2015-01-01

    The degree of sialylation of therapeutic glycoproteins affects its circulatory half-life and efficacy because incompletely sialylated glycoproteins are cleared from circulation by asialoglycoprotein receptors present in the liver cells. Mammalian expression systems, often employed in the production of these glycoprotein drugs, produce heterogeneously sialylated products. Here, we describe how to produce highly sialylated glycoproteins using a Chinese hamster ovary (CHO) cell glycosylation mutant called CHO-gmt4 with human erythropoietin (EPO) as a model glycoprotein. The protocol describes how to isolate and characterize the CHO glycosylation mutants and how to assess the sialylation of the recombinant protein using isoelectric focusing (IEF). It further describes how to inactivate the dihydrofolate reductase (DHFR) gene in these cells using zinc finger nuclease (ZFN) technology to enable gene amplification and the generation of stable cell lines producing highly sialylated EPO.

  3. Analysis of the trade-off between high crop yield and low yield instability at the global scale

    NASA Astrophysics Data System (ADS)

    Ben-Ari, Tamara; Makowski, David

    2016-10-01

    Yield dynamics of major crops species vary remarkably among continents. Worldwide distribution of cropland influences both the expected levels and the interannual variability of global yields. An expansion of cultivated land in the most productive areas could theoretically increase global production, but also increase global yield instability if the most productive regions are characterized by high interannual yield variability. In this letter, we use portfolio analysis to quantify the tradeoff between the expected values and the interannual variance of global yield. We compute optimal frontiers for four crop species i.e., maize, rice, soybean and wheat and show how the distribution of cropland among large world regions can be optimized to either increase expected global crop production or decrease its interannual variability. We also show that a preferential allocation of cropland in the most productive regions can increase global expected yield at the expense of yield stability. Theoretically, optimizing the distribution of a small fraction of total cultivated areas can help find a good compromise between low instability and high crop yields at the global scale.

  4. Selection of a Mutant of Escherichia coli Which Has High Mutation Rates

    PubMed Central

    Helling, Robert B.

    1968-01-01

    A mutation which causes high mutation rates in all other loci tested was induced with nitrosoguanidine and was selected through the ability of the progeny of such mutant cells to mutate to streptomycin resistance at a higher rate than the wild-type cells. This mutation (mut-2) and the Treffers' mutation (mutT1) mapped at approximately the same position to the right of leu. Specificity studies showed that the two mutations differ in rates of mutation produced. PMID:4879569

  5. Microvascular endothelium and pericytes: high yield, low passage cultures.

    PubMed

    Carson, M P; Haudenschild, C C

    1986-06-01

    Cultured microvascular endothelial cells (MEC) have become a valuable model for studies of microvascular physiology and pathology. Most current techniques involve manual removal of undesirable cell types or cloning, require one to several months, and yield high population doubling level cultures derived from a relatively small sample of the original population. We have devised a technique to more rapidly produce larger numbers of MEC. This method provided primary cultures consisting predominantly of MEC within 1 wk. The technique involves selective aspiration of gray matter from the bovine cerebral cortex followed by homogenization, sieving, enzymatic dissociation, and then dense plating (10(4) to 10(5) vessel fragments/cm2) onto gelatin- or fibronectin-coated plastic. Typical yields were 0.1 to 0.5 X 10(6) fragments/g of aspirated gray matter. The optimal culture medium for these cells was 15% equine plasma derived serum, 20% conditioned medium, 2% retinal extract, 60% fresh medium, and 500 micrograms/ml heparin. Cells attached within 24 h, well-spread colonies were present within 1 to 2 d, and cultures approached confluence within 2 to 3 d. Alkaline phosphatase staining confirmed the microvascular origin of the material plated. Morphology, Factor VIII-related antigen staining and 1,1'-dioctacecyl-3,3,3'3,-tetramethyl-indocarbocyanine perchlorate acetylated low density lipoprotein uptake suggested that MEC predominated. Cultures could be passaged and additionally purified by sequential exposure to pancreatin and trypsin-EDTA. Pancreatin selectively removed MEC colonies leaving a relatively homogeneous pericyte population. The relative ease with which such cultures can be produced should facilitate the in vitro study of brain microvascular function and may also provide insights useful for growing MEC from other vascular beds.

  6. High yield, single crystal ice via the Bridgman method

    NASA Astrophysics Data System (ADS)

    Bisson, Patrick; Groenzin, Henning; Barnett, Irene Li; Shultz, Mary Jane

    2016-03-01

    The surface chemistry of ice and of water is an important topic of study, especially given the role of ice and water in shaping the environment. Although snow, granular, and polycrystalline ice are often used in research, there are applications where large surface areas of a known crystallographic plane are required. For example, fundamental spectroscopy or scattering studies rely on large area samples of known crystalline orientation. In addition, due to its slower dynamics and decreased number of molecular configurations, ice can be viewed as a reduced complexity model for the complex hydrogen bonding environment found at the surface and within the bulk of liquid water. In our studies using Sum Frequency Generation (SFG) vibrational spectroscopy, we have shown that each crystalline face has a unique spectral signature and therefore a unique chemistry and chemical activity. A reliable, reproducible, high performance method of producing large single crystal samples is needed to support this surface chemistry research. The design, construction, and use of a computer-controlled, ice-growth machine based on the Stockbarger modified Bridgeman technique is described. The instrument reliably produces relatively large single crystals that are optically flawless (that is, no visible flaws when viewed in a crossed polarizer), and in very high yield. Success rates of 95% are typical. Such performance has not been observed in the literature.

  7. Analysis on the DNA Fingerprinting of Aspergillus Oryzae Mutant Induced by High Hydrostatic Pressure

    NASA Astrophysics Data System (ADS)

    Wang, Hua; Zhang, Jian; Yang, Fan; Wang, Kai; Shen, Si-Le; Liu, Bing-Bing; Zou, Bo; Zou, Guang-Tian

    2011-01-01

    The mutant strains of aspergillus oryzae (HP300a) are screened under 300 MPa for 20 min. Compared with the control strains, the screened mutant strains have unique properties such as genetic stability, rapid growth, lots of spores, and high protease activity. Random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) are used to analyze the DNA fingerprinting of HP300a and the control strains. There are 67.9% and 51.3% polymorphic bands obtained by these two markers, respectively, indicating significant genetic variations between HP300a and the control strains. In addition, comparison of HP300a and the control strains, the genetic distances of random sequence and simple sequence repeat of DNA are 0.51 and 0.34, respectively.

  8. Arabidopsis hot mutants define multiple functions required for acclimation to high temperatures.

    PubMed

    Hong, Suk-Whan; Lee, Ung; Vierling, Elizabeth

    2003-06-01

    Plants acquire thermotolerance to lethal high temperatures if first exposed to moderately high temperature or if temperature is increased gradually to an otherwise lethal temperature. We have taken a genetic approach to dissecting acquired thermotolerance by characterizing loss-of-function thermotolerance mutants in Arabidopsis. In previous work, we identified single recessive alleles of four loci required for thermotolerance of hypocotyl elongation, hot1-1, hot2-1, hot3-1, and hot4-1. Completed screening of M2 progeny from approximately 2500 M1 plants has now identified new alleles of three of these original loci, along with three new loci. The low mutant frequency suggests that a relatively small number of genes make a major contribution to this phenotype or that other thermotolerance genes encode essential or redundant functions. Further analysis of the original four loci was performed to define the nature of their thermotolerance defects. Although the HOT1 locus was shown previously to encode a major heat shock protein (Hsp), Hsp101, chromosomal map positions indicate that HOT2, 3, and 4 do not correspond to major Hsp or heat shock transcription factor genes. Measurement of thermotolerance at different growth stages reveals that the mutants have growth stage-specific heat sensitivity. Analysis of Hsp accumulation shows that hot2 and hot4 produce normal levels of Hsps, whereas hot3 shows reduced accumulation. Thermotolerance of luciferase activity and of ion leakage also varies in the mutants. These data provide the first direct genetic evidence, to our knowledge, that distinct functions, independent of Hsp synthesis, are required for thermotolerance, including protection of membrane integrity and recovery of protein activity/synthesis. PMID:12805605

  9. High-Yield Synthesis of Stoichiometric Boron Nitride Nanostructures

    DOE PAGESBeta

    Nocua, José E.; Piazza, Fabrice; Weiner, Brad R.; Morell, Gerardo

    2009-01-01

    Boron nimore » tride (BN) nanostructures are structural analogues of carbon nanostructures but have completely different bonding character and structural defects. They are chemically inert, electrically insulating, and potentially important in mechanical applications that include the strengthening of light structural materials. These applications require the reliable production of bulk amounts of pure BN nanostructures in order to be able to reinforce large quantities of structural materials, hence the need for the development of high-yield synthesis methods of pure BN nanostructures. Using borazine ( B 3 N 3 H 6 ) as chemical precursor and the hot-filament chemical vapor deposition (HFCVD) technique, pure BN nanostructures with cross-sectional sizes ranging between 20 and 50 nm were obtained, including nanoparticles and nanofibers. Their crystalline structure was characterized by (XRD), their morphology and nanostructure was examined by (SEM) and (TEM), while their chemical composition was studied by (EDS), (FTIR), (EELS), and (XPS). Taken altogether, the results indicate that all the material obtained is stoichiometric nanostructured BN with hexagonal and rhombohedral crystalline structure.« less

  10. Photoprotection mutants of Arabidopsis thaliana acclimate to high light by increasing photosynthesis and specific antioxidants.

    PubMed

    Golan, Talila; Müller-Moulé, Patricia; Niyogi, Krishna K

    2006-05-01

    Biochemical and physiological acclimation to different light environments is crucial for plant growth and survival. In high light (HL), feedback de-excitation (qE) is a well-known photoprotective mechanism that dissipates excess excitation energy in the light-harvesting antenna of photosystem II (PSII) and relieves excitation pressure in the photosynthetic electron transport chain. The xanthophylls zeaxanthin (Z) and lutein (L) function in qE, but also have roles as antioxidants. Although several studies have shown that qE is important during short-term fluctuations in light intensity, here we show that it is not required for the growth of Arabidopsis thaliana in prolonged HL conditions in the laboratory. Mutants that are deficient in qE alone, qE and Z synthesis, or in qE, Z synthesis and also L synthesis were able to grow at 1800 micromol photons m(-2) s(-1) and exhibited no major symptoms of photooxidative stress. The mutants (and wild type) acclimated to HL by increasing photosynthetic capacity and decreasing light harvesting, which together rendered qE less important for photoprotection. At a metabolite level, the HL-grown mutants appeared to compensate for their remaining qE deficit with increased alpha-tocopherol and ascorbate levels compared to the wild type. The specificity of this response provides insight into the relationship between qE and the antioxidant network in plants.

  11. Lichen Symbiosis: Nature's High Yielding Machines for Induced Hydrogen Production

    PubMed Central

    Papazi, Aikaterini; Kastanaki, Elizabeth; Pirintsos, Stergios; Kotzabasis, Kiriakos

    2015-01-01

    Hydrogen is a promising future energy source. Although the ability of green algae to produce hydrogen has long been recognized (since 1939) and several biotechnological applications have been attempted, the greatest obstacle, being the O2-sensitivity of the hydrogenase enzyme, has not yet been overcome. In the present contribution, 75 years after the first report on algal hydrogen production, taking advantage of a natural mechanism of oxygen balance, we demonstrate high hydrogen yields by lichens. Lichens have been selected as the ideal organisms in nature for hydrogen production, since they consist of a mycobiont and a photobiont in symbiosis. It has been hypothesized that the mycobiont’s and photobiont’s consumption of oxygen (increase of COX and AOX proteins of mitochondrial respiratory pathways and PTOX protein of chrolorespiration) establishes the required anoxic conditions for the activation of the phycobiont’s hydrogenase in a closed system. Our results clearly supported the above hypothesis, showing that lichens have the ability to activate appropriate bioenergetic pathways depending on the specific incubation conditions. Under light conditions, they successfully use the PSII-dependent and the PSII-independent pathways (decrease of D1 protein and parallel increase of PSaA protein) to transfer electrons to hydrogenase, while under dark conditions, lichens use the PFOR enzyme and the dark fermentative pathway to supply electrons to hydrogenase. These advantages of lichen symbiosis in combination with their ability to survive in extreme environments (while in a dry state) constitute them as unique and valuable hydrogen producing natural factories and pave the way for future biotechnological applications. PMID:25826211

  12. Lichen symbiosis: nature's high yielding machines for induced hydrogen production.

    PubMed

    Papazi, Aikaterini; Kastanaki, Elizabeth; Pirintsos, Stergios; Kotzabasis, Kiriakos

    2015-01-01

    Hydrogen is a promising future energy source. Although the ability of green algae to produce hydrogen has long been recognized (since 1939) and several biotechnological applications have been attempted, the greatest obstacle, being the O2-sensitivity of the hydrogenase enzyme, has not yet been overcome. In the present contribution, 75 years after the first report on algal hydrogen production, taking advantage of a natural mechanism of oxygen balance, we demonstrate high hydrogen yields by lichens. Lichens have been selected as the ideal organisms in nature for hydrogen production, since they consist of a mycobiont and a photobiont in symbiosis. It has been hypothesized that the mycobiont's and photobiont's consumption of oxygen (increase of COX and AOX proteins of mitochondrial respiratory pathways and PTOX protein of chrolorespiration) establishes the required anoxic conditions for the activation of the phycobiont's hydrogenase in a closed system. Our results clearly supported the above hypothesis, showing that lichens have the ability to activate appropriate bioenergetic pathways depending on the specific incubation conditions. Under light conditions, they successfully use the PSII-dependent and the PSII-independent pathways (decrease of D1 protein and parallel increase of PSaA protein) to transfer electrons to hydrogenase, while under dark conditions, lichens use the PFOR enzyme and the dark fermentative pathway to supply electrons to hydrogenase. These advantages of lichen symbiosis in combination with their ability to survive in extreme environments (while in a dry state) constitute them as unique and valuable hydrogen producing natural factories and pave the way for future biotechnological applications.

  13. Lichen symbiosis: nature's high yielding machines for induced hydrogen production.

    PubMed

    Papazi, Aikaterini; Kastanaki, Elizabeth; Pirintsos, Stergios; Kotzabasis, Kiriakos

    2015-01-01

    Hydrogen is a promising future energy source. Although the ability of green algae to produce hydrogen has long been recognized (since 1939) and several biotechnological applications have been attempted, the greatest obstacle, being the O2-sensitivity of the hydrogenase enzyme, has not yet been overcome. In the present contribution, 75 years after the first report on algal hydrogen production, taking advantage of a natural mechanism of oxygen balance, we demonstrate high hydrogen yields by lichens. Lichens have been selected as the ideal organisms in nature for hydrogen production, since they consist of a mycobiont and a photobiont in symbiosis. It has been hypothesized that the mycobiont's and photobiont's consumption of oxygen (increase of COX and AOX proteins of mitochondrial respiratory pathways and PTOX protein of chrolorespiration) establishes the required anoxic conditions for the activation of the phycobiont's hydrogenase in a closed system. Our results clearly supported the above hypothesis, showing that lichens have the ability to activate appropriate bioenergetic pathways depending on the specific incubation conditions. Under light conditions, they successfully use the PSII-dependent and the PSII-independent pathways (decrease of D1 protein and parallel increase of PSaA protein) to transfer electrons to hydrogenase, while under dark conditions, lichens use the PFOR enzyme and the dark fermentative pathway to supply electrons to hydrogenase. These advantages of lichen symbiosis in combination with their ability to survive in extreme environments (while in a dry state) constitute them as unique and valuable hydrogen producing natural factories and pave the way for future biotechnological applications. PMID:25826211

  14. A high-throughput method for isolation of salicylic acid metabolic mutants

    PubMed Central

    2010-01-01

    Background Salicylic acid (SA) is a key defense signal molecule against biotrophic pathogens in plants. Quantification of SA levels in plants is critical for dissecting the SA-mediated immune response. Although HPLC and GC/MS are routinely used to determine SA concentrations, they are expensive and time-consuming. We recently described a rapid method for a bacterial biosensor Acinetobacter sp. ADPWH_lux-based SA quantification, which enables high-throughput analysis. In this study we describe an improved method for fast sample preparation, and present a high-throughput strategy for isolation of SA metabolic mutants. Results On the basis of the previously described biosensor-based method, we simplified the tissue collection and the SA extraction procedure. Leaf discs were collected and boiled in Luria-Bertani (LB), and then the released SA was measured with the biosensor. The time-consuming steps of weighing samples, grinding tissues and centrifugation were avoided. The direct boiling protocol detected similar differences in SA levels among pathogen-infected wild-type, npr1 (nonexpressor of pathogenesis-related genes), and sid2 (SA induction-deficient) plants as did the previously described biosensor-based method and an HPLC-based approach, demonstrating the efficacy of the protocol presented here. We adapted this protocol to a high-throughput format and identified six npr1 suppressors that accumulated lower levels of SA than npr1 upon pathogen infection. Two of the suppressors were found to be allelic to the previously identified eds5 mutant. The other four are more susceptible than npr1 to the bacterial pathogen Pseudomonas syringae pv. maculicola ES4326 and their identity merits further investigation. Conclusions The rapid SA extraction method by direct boiling of leaf discs further reduced the cost and time required for the biosensor Acinetobacter sp. ADPWH_lux-based SA estimation, and allowed the screening for npr1 suppressors that accumulated less SA than npr1

  15. High level of antibiotic production in a double polyphosphate kinase and phosphate-binding protein mutant of Streptomyces lividans.

    PubMed

    Díaz, Margarita; Sevillano, Laura; Rico, Sergio; Lombo, Felipe; Braña, Alfredo F; Salas, Jose A; Mendez, Carmen; Santamaría, Ramón I

    2013-05-01

    Phosphate metabolism regulates most of the life processes of microorganisms. In the present work we obtained and studied a Streptomyces lividans ppk/pstS double mutant, which lacks polyphosphate kinase (PPK) and the high-affinity phosphate-binding protein (PstS), impairing at the same time the intracellular storage of polyphosphate and the intake of new inorganic phosphate from a phosphate-limited medium, respectively. In some of the aspects analyzed, the ppk/pstS double mutant was more similar to the wt strain than was the single pstS mutant. The double mutant was thus able to grow in phosphate-limited media, whereas the pstS mutant required the addition of 1 mM phosphate under the assay conditions used. The double mutant was able to incorporate more than one fourth of the inorganic phosphate incorporated by the wt strain, whereas phosphate incorporation was almost completely impaired in the pstS mutant. Noteworthy, under phosphate limitation conditions, the double ppk/pstS mutant showed a higher production of the endogenous antibiotic actinorhodin and the heterologous antitumor 8-demethyl-tetracenomycin (up to 10-fold with respect to the wt strain), opening new possibilities for the use of this strain in the heterologous expression of antibiotic pathways.

  16. Are the yields of major cereal crops stagnating? Results from the newly developed high spatial resolution crop yield time series

    NASA Astrophysics Data System (ADS)

    Ray, D. K.; Ramankutty, N.; Foley, J. A.

    2011-12-01

    A variety of global scale studies that use crop yield time series for the last 50 years have remained constrained to using national level information due to the lack of high spatial resolution crop yield time series data. In this presentation we will unveil a new global crop yield data set for the 1961-2008 time period, at 5 min spatial resolution, and covering 174 crops. We developed this data by collecting national and sub-national harvested area and production information for individual crops. This new dataset can be used to answer questions related to global agriculture at a resolution and over a time period not previously possible. We have used this new dataset to address the question of whether the yields of the three important cereal crops -- maize, rice and wheat -- are stagnating as widely reported. Our results show that while in the older crop belts of the world yield improvements have slowed, a green revolution type of major yield increases in maize, rice and wheat are continuing in newly cultivated areas of the world.

  17. Anomalous softening of yield strength in tantalum at high pressures

    SciTech Connect

    Jing, Qiumin Wu, Qiang; Xu, Ji-an; Bi, Yan; Liu, Lei; Liu, Shenggang; Zhang, Yi; Geng, Huayun

    2015-02-07

    The pressure dependence of the yield strength of tantalum was investigated experimentally up to 101 GPa at room temperature using a diamond anvil cell. A yield strength softening is observed between 52 and 84 GPa, whereas a normal trend is observed below 52 GPa and above 84 GPa. The onset pressure of the softening is in agreement with previous results obtained by the pressure gradient method and shock wave experiments. This unusual strength softening in tantalum is not related with structural transformation, preferred orientation, or material damage. Our measurements indicate that microscopic deviatoric strain is the major reason for the observed strength softening in tantalum.

  18. Jasmonates induce nonapoptotic death in high-resistance mutant p53-expressing B-lymphoma cells.

    PubMed

    Fingrut, Orit; Reischer, Dorit; Rotem, Ronit; Goldin, Natalia; Altboum, Irit; Zan-Bar, Israel; Flescher, Eliezer

    2005-11-01

    Mutations in p53, a tumor suppressor gene, occur in more than half of human cancers. Therefore, we tested the hypothesis that jasmonates (novel anticancer agents) can induce death in mutated p53-expressing cells. Two clones of B-lymphoma cells were studied, one expressing wild-type (wt) p53 and the other expressing mutated p53. Jasmonic acid and methyl jasmonate (0.25-3 mM) were each equally cytotoxic to both clones, whereas mutant p53-expressing cells were resistant to treatment with the radiomimetic agent neocarzinostatin and the chemotherapeutic agent bleomycin. Neocarzinostatin and bleomycin induced an elevation in the p53 levels in wt p53-expressing cells, whereas methyl jasmonate did not. Methyl jasmonate induced mostly apoptotic death in the wt p53-expressing cells, while no signs of early apoptosis were detected in mutant p53-expressing cells. In contrast, neocarzinostatin and bleomycin induced death only in wt p53-expressing cells, in an apoptotic mode. Methyl jasmonate induced a rapid depletion of ATP in both clones. In both clones, oligomycin (a mitochondrial ATP synthase inhibitor) did not increase ATP depletion induced by methyl jasmonate, whereas inhibition of glycolysis with 2-deoxyglucose did. High glucose levels protected both clones from methyl jasmonate-induced ATP depletion (and reduced methyl jasmonate-induced cytotoxicity), whereas high levels of pyruvate did not. These results suggest that methyl jasmonate induces ATP depletion mostly by compromising oxidative phosphorylation in the mitochondria. In conclusion, jasmonates can circumvent the resistance of mutant p53-expressing cells towards chemotherapy by inducing a nonapoptotic cell death.

  19. Metabolic engineering for high yielding L(-)-carnitine production in Escherichia coli

    PubMed Central

    2013-01-01

    Background L(-)-carnitine production has been widely studied because of its beneficial properties on various diseases and dysfunctions. Enterobacteria possess a specific biotransformation pathway which can be used for the enantioselective production of L(-)-carnitine. Although bioprocesses catalyzed by enzymes or whole cells can overcome the lack of enantioselectivity of chemical methods, current processes for L(−)-carnitine production still have severe disadvantages, such as the low yields, side reactions and the need of high catalyst concentrations and anaerobic conditions for proper expression of the biotransformation pathway. Additionally, genetically engineered strains so far constructed for L(-)-carnitine production are based on plasmids and, therefore, suffer from segregational unstability. Results In this work, a stable, high yielding strain for L(-)-carnitine production from low cost substrates was constructed. A metabolic engineering strategy was implemented in a multiple mutant for use in both growing and resting cells systems. The effect of mutations on gene expression and metabolism was analyzed to characterize the productivity constraints of the wild type and the overproducer strains. Precise deletion of genes which encode proteins of central and carnitine metabolisms were performed. Specifically, flux through the TCA cycle was increased by deletion of aceK (which encodes a bifunctional kinase/phosphatase which inhibits isocitrate dehydrogenase activity) and the synthesis of the by-product γ-butyrobetaine was prevented by deletion of caiA (which encodes a crotonobetainyl-CoA reductase). Both mutations led to improve the L(-)-carnitine production by 20 and 42%, respectively. Moreover, the highly regulated promoter of the cai operon was substituted by a constitutive artificial promoter increasing the biotransformation rate, even under aerobic conditions. Resting cells of the BW ΔaceK ΔcaiA p37cai strain produced 59.6 mmol l-1 · h-1 of L

  20. High molecular weight complexes of mutant superoxide dismutase 1: age-dependent and tissue-specific accumulation.

    PubMed

    Wang, Jiou; Xu, Guilian; Borchelt, David R

    2002-03-01

    Mutations in the cytosolic enzyme, superoxide dismutase 1, have been identified as the cause of motor neuron disease in a subset of cases of familial amyotrophic lateral sclerosis. It has been postulated that the injurious property of mutant enzyme resides in its propensity to aggregate or its propensity to catalyze deleterious, copper-mediated, chemistries. Aggregates of SOD1 have been identified, histologically, in neurons and astroglia of the spinal cords of SOD1-linked FALS patients and in transgenic mice that express these mutant proteins. In the present study, we have employed a technique used in detecting and quantifying aggregates of mutant huntingtin (cellulose acetate filtration) to examine the molecular characteristics of mutant SOD1 in three previously characterized transgenic mouse models of FALS. We show that the brains and spinal cords of these mice accumulate mutant SOD1 complexes that can be trapped by cellulose acetate filtration. The relative abundance of these structures increases dramatically with age. Although expressed to the same level in nonnervous tissues, mutant SOD1 was not found in high molecular weight structures. We conclude that some aspect of the biology of neural tissues (in a setting of declining motor neuron function) predisposes to the accumulation of high molecular weight complexes of mutant SOD1.

  1. Advanced scheme for high-yield laser driven nuclear reactions

    NASA Astrophysics Data System (ADS)

    Margarone, D.; Picciotto, A.; Velyhan, A.; Krasa, J.; Kucharik, M.; Mangione, A.; Szydlowsky, A.; Malinowska, A.; Bertuccio, G.; Shi, Y.; Crivellari, M.; Ullschmied, J.; Bellutti, P.; Korn, G.

    2015-01-01

    The use of a low contrast nanosecond laser pulse with a relatively low intensity (3  ×  1016 W cm-2) allowed the enhancing of the yield of induced nuclear reactions in advanced solid targets. In particular the ‘ultraclean’ proton-boron fusion reaction, producing energetic alpha particles without neutron generation, was chosen. A spatially well-defined layer of boron dopants in a hydrogen-enriched silicon substrate was used as a target. A combination of the specific target composition and the laser pulse temporal shape allowed the enhancing of the yield of alpha particles up to 109 per steradian. This result can be ascribed to the interaction of the long-laser pre-pulse with the target and to the optimal target geometry and composition.

  2. A high yield neutron target for cancer therapy

    NASA Technical Reports Server (NTRS)

    Alger, D. L.; Steinberg, R.

    1972-01-01

    A rotating target was developed that has the potential for providing an initial yield of 10 to the 13th power neutrons per second by the T(d,n)He-4 reaction, and a useable lifetime in excess of 600 hours. This yield and lifetime are indicated for a 300 Kv and 30 mA deuteron accelerator and a 30 microns thick titanium tritide film formed of the stoichiometric compound TiT2. The potential for extended lifetime is made possible by incorporating a sputtering electrode that permits use of titanium tritide thicknesses much greater than the deuteron range. The electrode is used to remove in situ depleted titanium layers to expose fresh tritide beneath. The utilization of the rotating target as a source of fast neutrons for cancer therapy is discussed.

  3. Anisotropic yielding of rocks at high temperatures and pressures

    SciTech Connect

    Kronenberg, A.K.; Russell, J.E.; Handin, J.; Gottschalk, R.R.; Shea, W.T.

    1987-12-01

    Results to date are: All of the starting materials for the three year project have been collected. Included in our collection are relatively fine-grained, fresh, oriented blocks of schist, gneiss, and micaceous quartzite with well-defined foliations and lineations as well as granite blocks oriented with respect to the principal quarrying orientations, the rift, grain, and hardway. A suite of samples has also been collected from an exposed granite stock and surrounding country rocks in order to evaluate the strengths and distribution of fabrics which may be encountered while drilling. These fabrics appear to be directly related to the forceful emplacement of the pluton. The literature on the mechanics of intrusion has been reviewed with regard to strain gradients and foliation development associated with diapiric flow. This information will be used to evaluate flow of varying fabrics on yield criteria within and surrounding magma chambers. Twenty-three successful experiments have been performed on samples of gneiss cored along six different orientations at temperatures ranging from 25{degrees} to 700{degrees}C. These experiments include extension tests, unconfined compression tests, and compression tests performed at P{sub c} = 100 MPa. Theoretical yield conditions for anisotropic materials have been reviewed and the assumptions upon which they are based probed. These yield conditions will ultimately be used to fit our data on gneiss, and the other foliated rocks under investigation. Two abstracts have been published and oral presentations made at the 1987 Fall Meeting of the American Geophysical Union, based upon our previous DOE-sponsored work on tensile fracturing of quartzite and related work on semi-brittle deformation of granitic rocks. 21 refs., 12 figs., 2 tabs.

  4. High yield neutron generator based on a high-current gasdynamic electron cyclotron resonance ion source

    SciTech Connect

    Skalyga, V.; Sidorov, A.; Izotov, I.; Golubev, S.; Razin, S.; Strelkov, A.; Tarvainen, O.; Koivisto, H.; Kalvas, T.

    2015-09-07

    In present paper, an approach for high yield compact D-D neutron generator based on a high current gasdynamic electron cyclotron resonance ion source is suggested. Results on dense pulsed deuteron beam production with current up to 500 mA and current density up to 750 mA/cm{sup 2} are demonstrated. Neutron yield from D{sub 2}O and TiD{sub 2} targets was measured in case of its bombardment by pulsed 300 mA D{sup +} beam with 45 keV energy. Neutron yield density at target surface of 10{sup 9} s{sup −1} cm{sup −2} was detected with a system of two {sup 3}He proportional counters. Estimations based on obtained experimental results show that neutron yield from a high quality TiD{sub 2} target bombarded by D{sup +} beam demonstrated in present work accelerated to 100 keV could reach 6 × 10{sup 10} s{sup −1} cm{sup −2}. It is discussed that compact neutron generator with such characteristics could be perspective for a number of applications like boron neutron capture therapy, security systems based on neutron scanning, and neutronography.

  5. High-yield microfabrication process for biomimetic artificial haircell sensors

    NASA Astrophysics Data System (ADS)

    Li, Junjun; Fan, Zhifang; Chen, Jack; Zou, Jun; Liu, Chang

    2002-07-01

    We have developed artificial haircell sensors based on an improved plastic deformation magnetic assembly (PDMA) method. The PDMA method is uniquely capable of realizing 3D microstructures in an efficient, IC-compatible manner. We have improved the design of the PDMA structure to significantly increase the yield by eliminating stress concentration. The robustness of the fabricated devices is enhanced using conformal Parylene deposition. The PDMA process uses magnetic force to bend surface micromachiend cantilever beams. If the bending is significant, the beams would enter plastic deformation regime and maintains permanently bent position even after the magnetic field is removed. Previously, the PDMA process is associated with appreciable yield loss during het magnetic assembly. The failure mechanisms is believed to be stress concentration at sharp corners defined by edges of sacrificial layers. In the new design, a structure enhancement feature is added to the surface micromachiend assembly and hence eliminating any sharp features. Haircells sensors are modular sensory units widely found in the biological world. Haircells are responsible for hearing, flow sensing, and for balancing/equilibrium. We are developing artificial haircell sensors using advanced micromachining technology. The artificial haircell sensors have similar physical scale compared with biological ones.

  6. Using the Nova target chamber for high-yield targets

    SciTech Connect

    Pitts, J.H.

    1987-09-28

    The existing 2.2-m-radius Nova aluminum target chamber, coated and lined with boron-seeded carbon shields, is proposed for use with 1000-MJ-yield targets in the next laser facility. The laser beam and diagnostic holes in the target chamber are left open and the desired 10/sup -2/ Torr vacuum is maintained both inside and outside the target chamber; a larger target chamber room is the vacuum barrier to the atmosphere. The hole area available is three times that necessary to maintain a maximum fluence below 12 J/cm/sup 2/ on optics placed at a radius of 10 m. Maximum stress in the target chamber wall is 73 MPa, which complies with the intent of the ASME Pressure Vessel Code. However, shock waves passing through the inner carbon shield could cause it to comminute. We propose tests and analyses to ensure that the inner carbon shield survives the environment. 13 refs.

  7. Gluconacetobacter hansenii subsp. nov., a high-yield bacterial cellulose producing strain induced by high hydrostatic pressure.

    PubMed

    Ge, Han-Jing; Du, Shuang-Kui; Lin, De-Hui; Zhang, Jun-Na; Xiang, Jin-Le; Li, Zhi-Xi

    2011-12-01

    Strain M(438), deposited as CGMCC3917 and isolated from inoculums of bacterial cellulose (BC) producing strain screened in homemade vinegar and then induced by high hydrostatic pressure treatment (HHP), has strong ability to produce BC more than three times as that of its initial strain. It is the highest yield BC-producing strain ever reported. In this paper, M(438) was identidied as Gluconacetobacter hansenii subsp. nov. on the basis of the results obtained by examining it phylogenetically, phenotypically, and physiologically-biochemically. Furthermore, the genetic diversity of strain M(438) and its initial strain was examined by amplified fragment length polymorphism. The results indicated that strain M(438) was a deletion mutant induced by HHP, and the only deleted sequence showed 99% identity with 24,917-24,723 bp in the genome sequence of Ga. hansenii ATCC23769, and the complement gene sequence was at 24,699-25,019 bp with local tag GXY_15142, which codes small multidrug resistance (SMR) protein. It can be inferred that SMR might be related to inhibiting BC production to a certain extent.

  8. Anistotropic yielding of rocks at high temperatures and pressures

    SciTech Connect

    Kronenberg, A.K.; Russell, J.E.; Carter, N.L.

    1990-10-14

    The anisotropic deformation of foliated and linealed rocks has been investigated, primarily to predict the mechanical response of rocks surrounding buried magma chambers to the stress fields generated by deep drilling. The principal application in this regard has been to evaluate, the scientific feasibility of extracting geothermal energy from buried magma chambers. Our approach has been to perform triaxial extension and compression tests at temperatures and pressures representative of the borehole environment on samples cored along six selected orientations and to fit the data to an orthohombric yield criterion. We have investigated Four-Mile gneiss (a strongly layered gneiss with well defined lineation), a biotite-rich schist, and Westerly granite (using a block oriented with respect to the granite's rift, grain, and hardway). Progress has been made in three areas: the experimental determination of strength anisotropies for the three starting materials, theoretical treatment and modeling of the results, and characterization of fabrics surrounding magma bodies resulting from their diaperic emplacement into shallow portions of the Earth's crust. In addition, results have been obtained for the tensile fracture of quartzite, basal slip and anisotropy of biotite single crystals, and anisotropic flow of bedded rocksalt.

  9. Separating heat stress from moisture stress: analyzing yield response to high temperature in irrigated maize

    NASA Astrophysics Data System (ADS)

    Carter, Elizabeth K.; Melkonian, Jeff; Riha, Susan J.; Shaw, Stephen B.

    2016-09-01

    Several recent studies have indicated that high air temperatures are limiting maize (Zea mays L.) yields in the US Corn Belt and project significant yield losses with expected increases in growing season temperatures. Further work has suggested that high air temperatures are indicative of high evaporative demand, and that decreases in maize yields which correlate to high temperatures and vapor pressure deficits (VPD) likely reflect underlying soil moisture limitations. It remains unclear whether direct high temperature impacts on yields, independent of moisture stress, can be observed under current temperature regimes. Given that projected high temperature and moisture may not co-vary the same way as they have historically, quantitative analyzes of direct temperature impacts are critical for accurate yield projections and targeted mitigation strategies under shifting temperature regimes. To evaluate yield response to above optimum temperatures independent of soil moisture stress, we analyzed climate impacts on irrigated maize yields obtained from the National Corn Growers Association (NCGA) corn yield contests for Nebraska, Kansas and Missouri. In irrigated maize, we found no evidence of a direct negative impact on yield by daytime air temperature, calculated canopy temperature, or VPD when analyzed seasonally. Solar radiation was the primary yield-limiting climate variable. Our analyses suggested that elevated night temperature impacted yield by increasing rates of phenological development. High temperatures during grain-fill significantly interacted with yields, but this effect was often beneficial and included evidence of acquired thermo-tolerance. Furthermore, genetics and management—information uniquely available in the NCGA contest data—explained more yield variability than climate, and significantly modified crop response to climate. Thermo-acclimation, improved genetics and changes to management practices have the potential to partially or completely

  10. Generation and Multi-phenotypic High-content Screening of Coxiella burnetii Transposon Mutants

    PubMed Central

    Martinez, Eric; Cantet, Franck; Bonazzi, Matteo

    2015-01-01

    Invasion and colonization of host cells by bacterial pathogens depend on the activity of a large number of prokaryotic proteins, defined as virulence factors, which can subvert and manipulate key host functions. The study of host/pathogen interactions is therefore extremely important to understand bacterial infections and develop alternative strategies to counter infectious diseases. This approach however, requires the development of new high-throughput assays for the unbiased, automated identification and characterization of bacterial virulence determinants. Here, we describe a method for the generation of a GFP-tagged mutant library by transposon mutagenesis and the development of high-content screening approaches for the simultaneous identification of multiple transposon-associated phenotypes. Our working model is the intracellular bacterial pathogen Coxiellaburnetii, the etiological agent of the zoonosis Q fever, which is associated with severe outbreaks with a consequent health and economic burden. The obligate intracellular nature of this pathogen has, until recently, severely hampered the identification of bacterial factors involved in host pathogen interactions, making of Coxiella the ideal model for the implementation of high-throughput/high-content approaches. PMID:25992686

  11. High-yield anthocyanin biosynthesis in engineered Escherichia coli.

    PubMed

    Yan, Yajun; Li, Zhen; Koffas, Mattheos A G

    2008-05-01

    Anthocyanins are red, purple, or blue plant water-soluble pigments. In the past two decades, anthocyanins have received extensive studies for their anti-oxidative, anti-inflammatory, anti-cancer, anti-obesity, anti-diabetic, and cardioprotective properties. In the present study, anthocyanin biosynthetic enzymes from different plant species were characterized and employed for pathway construction leading from inexpensive precursors such as flavanones and flavan-3-ols to anthocyanins in Escherichia coli. The recombinant E. coli cells successfully achieved milligram level production of two anthocyanins, pelargonidin 3-O-glucoside (0.98 mg/L) and cyanidin 3-O-gluside (2.07 mg/L) from their respective flavanone precursors naringenin and eriodictyol. Cyanidin 3-O-glucoside was produced at even higher yields (16.1 mg/L) from its flavan-3-ol, (+)-catechin precursor. Further studies demonstrated that availability of the glucosyl donor, UDP-glucose, was the key metabolic limitation, while product instability at normal pH was also identified as a barrier for production improvement. Therefore, various optimization strategies were employed for enhancing the homogenous synthesis of UDP-glucose in the host cells while at the same time stabilizing the final anthocyanin product. Such optimizations included culture medium pH adjustment, the creation of fusion proteins and the rational manipulation of E. coli metabolic network for improving the intracellular UDP-glucose metabolic pool. As a result, production of pelargonidin 3-O-glucoside at 78.9 mg/L and cyanidin 3-O-glucoside at 70.7 mg/L was achieved from their precursor flavan-3-ols without supplementation with extracellular UDP-glucose. These results demonstrate the efficient production of the core anthocyanins for the first time and open the possibility for their commercialization for pharmaceutical and nutraceutical applications.

  12. High-risk energy plans yield low rewards

    NASA Astrophysics Data System (ADS)

    Gwynne, Peter

    2009-10-01

    Scientists have complained that a new research body run by the US Department of Energy (DOE) is suffering from management problems and is rejecting funding proposals without stating why. The DOE's Advanced Research Projects Agency-Energy (ARPA-E) - created in 2007 - is designed to fund high-risk, high-payoff research and development projects in energy. However, the agency, which received its first budget in April, has so far turned down almost 95% of proposals after the first round.

  13. Biomass Accretion and Yield of Erect Leafed and Conventional Sorghum at Low and High Population Densities

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two sorghum isolines, a wild type (BTx 623) and an erect leaf mutant line (ERL 20) isolated from the wild type were field grown in rectilinear arrays at low (25 plants m-2) and high (12 plants/m-2) densities with sub-surface drip irrigation in an effort to eliminate confounding drought effects. Cano...

  14. Enhancing growth rate and lipid yield of Chlorella with nuclear irradiation under high salt and CO2 stress.

    PubMed

    Cheng, Jun; Lu, Hongxiang; Huang, Yun; Li, Ke; Huang, Rui; Zhou, Junhu; Cen, Kefa

    2016-03-01

    In order to produce biodiesel from microalgae cultured with abundant seawater, Chlorella sp. was mutated with (137)Se-γ ray irradiation and domesticated with f/2 seawater culture medium (salinity=3 wt.%) under 15 vol.% CO2 stress. Biomass yield of the mutant increased by 25% compared with wild species and lipid content increased to 54.9%. When nitrogen and phosphorus concentrations in the initial substrate increased, the increased propagation speed of the mutant resulted in decreased cell diameter by 26.6% and decreased cell wall thickness by 69.7%. The dramatically increased biomass yield of the mutant with sufficient initial substrate and relative nitrogen starvation in the later growth period with continuous 15 vol.% CO2 led to an increased lipid yield of 1.0 g/L. The long-chain unsaturated fatty acids increased, whereas short-chain saturated fatty acids decreased.

  15. High School Student Physics Research Experience Yields Positive Results

    ERIC Educational Resources Information Center

    Podolak, K. R.; Walters, M. J.

    2016-01-01

    All high school students that wish to continue onto college are seeking opportunities to be competitive in the college market. They participate in extra-curricular activities which are seen to foster creativity and the skills necessary to do well in the college environment. In the case of students with an interest in physics, participating in a…

  16. High-biomass sorghum yield estimate with aerial imagery

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Abstract. To reach the goals laid out by the U.S. Government for displacing fossil fuels with biofuels, agricultural production of dedicated biomass crops is required. High-biomass sorghum is advantageous across wide regions because it requires less water per unit dry biomass and can produce very hi...

  17. High-Yield Metal Materials for Photocathode RF Gun

    SciTech Connect

    Wang, X.J.; Ben-Zvi, I.; Smedley, J.; Srinivasan-Rao, T.; Woodle, M.; Palmer, D.T.; Miller, R.H.

    1997-11-01

    Significant progress has been made in both improving the quantum efficiency and understanding the physics process of photoemission of metal photocathode materials under high electric field. Metal material, copper and magnesium were installed on the photocathode RF gun, and experimentally investigated using frequency quadrupled Nd:Yag laser (266 nm). A systemic procedure was developed for preparing cathode; this involves diamond polishing, ultrasonic cleaning and high temperature vacuum bake out. Using laser cleaning and explosive electron emission cleaning, the highest quantum efficiency measured for copper and magnesium are 0.02% and 0.3%, respectively. The Schottky effect was experimental investigated, the quantum efficiency of the Cu is dominated by the Schottky effect since the work function of the copper is almost equal to the photon energy used (4.66 eV). The quantum efficiency of the magnesium is significant enhanced by the Schottky effect.

  18. High school student physics research experience yields positive results

    NASA Astrophysics Data System (ADS)

    Podolak, K. R.; Walters, M. J.

    2016-03-01

    All high school students that wish to continue onto college are seeking opportunities to be competitive in the college market. They participate in extra-curricular activities which are seen to foster creativity and the skills necessary to do well in the college environment. In the case of students with an interest in physics, participating in a small scale research project while in high school gives them the hands on experience and ultimately prepares them more for the college experience. SUNY Plattsburgh’s Physics department started a five-week summer program for high school students in 2012. This program has proved not only beneficial for students while in the program, but also as they continue on in their development as scientists/engineers. Independent research, such as that offered by SUNY Plattsburgh’s five-week summer program, offers students a feel and taste of the culture of doing research, and life as a scientist. It is a short-term, risk free way to investigate whether a career in research or a particular scientific field is a good fit.

  19. Clickstream data yields high-resolution maps of science

    SciTech Connect

    Bollen, Johan; Van De Sompel, Herbert; Hagberg, Aric; Bettencourt, Luis; Chute, Ryan; Rodriguez, Marko A; Balakireva, Lyudmila

    2009-01-01

    Intricate maps of science have been created from citation data to visualize the structure of scientific activity. However, most scientific publications are now accessed online. Scholarly web portals record detailed log data at a scale that exceeds the number of all existing citations combined. Such log data is recorded immediately upon publication and keeps track of the sequences of user requests (clickstreams) that are issued by a variety of users across many different domains. Given these advantagees of log datasets over citation data, we investigate whether they can produce high-resolution, more current maps of science.

  20. Increase in Ty1 cDNA Recombination in Yeast sir4 Mutant Strains at High Temperature

    PubMed Central

    Radford, Sarah J.; Boyle, Meredith L.; Sheely, Catherine J.; Graham, Joel; Haeusser, Daniel P.; Zimmerman, Leigh; Keeney, Jill B.

    2004-01-01

    Transposition of the Ty1 element of the yeast Saccharomyces cerevisiae is temperature sensitive. We have identified a null allele of the silent information regulator gene SIR4 as a host mutant that allows for transposition at high temperature. We show that the apparent increase in transposition activity in sir4 mutant strains at high temperature is dependent on the RAD52 gene and is thus likely resulting from an increase in Ty1 cDNA recombination, rather than in IN-mediated integration. General cellular recombination is not increased at high temperature, suggesting that the increase in recombination at high temperature in sir4 mutants is specific for Ty1 cDNA. Additionally, this high-temperature Ty1 recombination was found to be dependent on functional Sir2p and Sir3p. We speculate that the increase in recombination seen in sir4 mutants at high temperature may be due to changes in chromatin structure or Ty1 interactions with chromosomal structures resulting in higher recombination rates. PMID:15454529

  1. High-Throughput FRET Assay Yields Allosteric SERCA Activators

    PubMed Central

    Cornea, Razvan L.; Lockamy, Elizabeth L.; Gruber, Simon J.; Muretta, Joseph M.; Jin, Dongzhu; Chen, Jiqiu; Dahl, Russell; Bartfai, Tamas; Zsebo, Krisztina M.; Gillispie, Gregory D.; Thomas, David D.

    2013-01-01

    Using fluorescence resonance energy transfer (FRET), we performed a high-throughput screen (HTS) in a reconstituted membrane system, seeking compounds that reverse inhibition of sarco-/endoplasmic reticulum Ca-ATPase (SERCA) by its endogenous regulator, phospholamban (PLB). Such compounds have long been sought to correct aberrant Ca2+ regulation in heart failure. Donor-SERCA was reconstituted in phospholipid membranes with or without acceptor-PLB, and FRET was measured in a steady-state fluorescence microplate reader. A 20,000-compound library was tested in duplicate. Compounds that decreased FRET by more than three standard deviations were considered hits. From 43 primary hits (0.2%), 31 (72%) were found to be false positives upon more thorough testing. The remaining 12 hits were tested in assays of Ca-ATPase activity, and six of these activated SERCA significantly, by as much as 60%, and several also enhanced cardiomyocyte contractility. These compounds directly activated SERCA from heart and other tissues. These results validate our FRET approach and set the stage for medicinal chemistry and pre-clinical testing. We were concerned about the high rate of false positives, resulting from the low precision of steady-state fluorescence. Preliminary studies with a novel fluorescence lifetime plate reader show 20-fold higher precision. This instrument can dramatically increase the quality of future HT. PMID:22923787

  2. High-throughput FRET assay yields allosteric SERCA activators.

    PubMed

    Cornea, Razvan L; Gruber, Simon J; Lockamy, Elizabeth L; Muretta, Joseph M; Jin, Dongzhu; Chen, Jiqiu; Dahl, Russell; Bartfai, Tamas; Zsebo, Krisztina M; Gillispie, Gregory D; Thomas, David D

    2013-01-01

    Using fluorescence resonance energy transfer (FRET), we performed a high-throughput screen (HTS) in a reconstituted membrane system, seeking compounds that reverse inhibition of sarcoplasmic reticulum Ca-ATPase (SERCA) by its cardiac regulator, phospholamban (PLB). Such compounds have long been sought to correct aberrant Ca(2+) regulation in heart failure. Donor-SERCA was reconstituted in phospholipid membranes with or without acceptor-PLB, and FRET was measured in a steady-state fluorescence microplate reader. A 20 000-compound library was tested in duplicate. Compounds that decreased FRET by more than three standard deviations were considered hits. From 43 hits (0.2%), 31 (72%) were found to be false-positives upon more thorough FRET testing. The remaining 12 hits were tested in assays of Ca-ATPase activity, and six of these activated SERCA significantly, by as much as 60%, and several also enhanced cardiomyocyte contractility. These compounds directly activated SERCA from heart and other tissues. These results validate our FRET approach and set the stage for medicinal chemistry and preclinical testing. We were concerned about the high rate of false-positives, resulting from the low precision of steady-state fluorescence. Preliminary studies with a novel fluorescence lifetime plate reader show 20-fold higher precision. This instrument can dramatically increase the quality of future HTS.

  3. Kinetics of high-Level of ß-glucosidase production by a 2-deoxyglucose-resistant mutant of Humicola lanuginosa in submerged fermentation

    PubMed Central

    Bokhari, Syed Ali Imran; Latif, Farooq; Rajoka, Muhammad Ibrahim

    2008-01-01

    A 2-deoxyglucose-resistant mutant (M7) of Humicola lanuginosa was obtained by exposing conidia to γ-rays and permitting expression in broth containing 0.6% 2-deoxyglucose (DG) and cellobiose (1%) before plating on DG esculin-ferric ammonium citrate agar medium from which colonies showing faster and bigger blackening zones were selected. Kinetic parameters for enhanced ß-glucosidase (BGL) synthesis by M7 were achieved when corncobs acted as the carbon source. The combination between corncobs and corn steep liquor was the best to support higher values of all product formation kinetic parameters. Effect of temperature on the kinetic and thermodynamic attributes of BGL production equilibrium in the wild organism and M7 was studied using batch process at eight different temperatures in shake-flask studies. The best performance was found at 45°C and 20 g L−1 corncobs in 64 h. Both growth and product formation (17.93 U mL−1) were remarkably high at 45°C and both were coupled under optimum working conditions. Product yield of BGL from the mutant M7 (1556.5 U g−1 dry corncobs) was significantly higher than the values reported on all fungal and bacterial systems. Mutation had thermo-stabilization influence on the organism and mutant required lower activation energy for growth and lower magnitudes of enthalpy and entropy for product formation than those demanded by the wild organism, other mesophilic and thermo-tolerant organisms. In the inactivation phase, the organisms needed lower values of activation energy, enthalpy and entropy for product formation equilibrium, confirming thermophilic nature of metabolic network possessed by the mutant organism. PMID:24031297

  4. A high yield method for growing primary canine keratinocytes.

    PubMed

    Watson, Adrian; Baker, Claire; Bailey, Julie; Fray, Tim; Markwell, Peter

    2004-07-01

    From a small amount of starting material, a large quantity of canine keratinocytes can be generated for experimental purposes using a refined method of explant culture to initiate the growth of basal cells with a high proliferative potential. The dividing capacity of cultures was promoted by a system selecting clonogenic cells onto an i3T3 feeder layer in combination with carefully monitoring cell morphology and passaging to select out excessive numbers of differentiated keratinocytes. Levels of contaminating dermal fibroblasts, which if left unchecked will overgrow keratinocytes, were kept to a minimum by a combination of careful explant micro-dissection to remove dermis, eliminating explants with signs of fibroblast growth as well as using cholera toxin, EGF and i3T3 feeder layers. The advantage of the method described is that it does not rely on the provision of large quantities of starting material thereby reducing the need for repeated tissue sampling, and passage numbers of five or six can be routinely achieved. This technique can therefore be useful to experimenters who require a regular and reliable source of cells for their studies. PMID:15158212

  5. Clickstream Data Yields High-Resolution Maps of Science

    PubMed Central

    Bollen, Johan; Van de Sompel, Herbert; Rodriguez, Marko A.; Balakireva, Lyudmila

    2009-01-01

    Background Intricate maps of science have been created from citation data to visualize the structure of scientific activity. However, most scientific publications are now accessed online. Scholarly web portals record detailed log data at a scale that exceeds the number of all existing citations combined. Such log data is recorded immediately upon publication and keeps track of the sequences of user requests (clickstreams) that are issued by a variety of users across many different domains. Given these advantages of log datasets over citation data, we investigate whether they can produce high-resolution, more current maps of science. Methodology Over the course of 2007 and 2008, we collected nearly 1 billion user interactions recorded by the scholarly web portals of some of the most significant publishers, aggregators and institutional consortia. The resulting reference data set covers a significant part of world-wide use of scholarly web portals in 2006, and provides a balanced coverage of the humanities, social sciences, and natural sciences. A journal clickstream model, i.e. a first-order Markov chain, was extracted from the sequences of user interactions in the logs. The clickstream model was validated by comparing it to the Getty Research Institute's Architecture and Art Thesaurus. The resulting model was visualized as a journal network that outlines the relationships between various scientific domains and clarifies the connection of the social sciences and humanities to the natural sciences. Conclusions Maps of science resulting from large-scale clickstream data provide a detailed, contemporary view of scientific activity and correct the underrepresentation of the social sciences and humanities that is commonly found in citation data. PMID:19277205

  6. Efficient production of a gene mutant cell line through integrating TALENs and high-throughput cell cloning.

    PubMed

    Sun, Changhong; Fan, Yu; Li, Juan; Wang, Gancheng; Zhang, Hanshuo; Xi, Jianzhong Jeff

    2015-02-01

    Transcription activator-like effectors (TALEs) are becoming powerful DNA-targeting tools in a variety of mammalian cells and model organisms. However, generating a stable cell line with specific gene mutations in a simple and rapid manner remains a challenging task. Here, we report a new method to efficiently produce monoclonal cells using integrated TALE nuclease technology and a series of high-throughput cell cloning approaches. Following this method, we obtained three mTOR mutant 293T cell lines within 2 months, which included one homozygous mutant line.

  7. Proteomic analysis of Spirogyra varians mutant with high starch content and growth rate induced by gamma irradiation.

    PubMed

    Yoon, Minchul; Choi, Jong-il; Kim, Gwang Hoon; Kim, Dong-Ho; Park, Don-Hee

    2013-06-01

    This study was conducted to develop a high-efficiency strain of Spirogyra varians for the production of biomass by radiation breeding. The characteristics of wild-type and mutant S. varians were analyzed through phenomenological and proteomic observations. The results of our phenomenological observations of the S. varians mutant demonstrated increases in growth rate and content of chlorophyll a, b, and a + b; in particular, a significant threefold increase was observed in starch accumulation. Proteomic analysis to investigate the differences in expression between wild-type and mutant proteins identified 18 proteins with significantly different expressions. From the literature review, it was confirmed that the up-regulated proteins were mainly involved in photosynthesis, carbohydrate biosynthesis, and energy metabolism. These results suggest the possibility of algae development by radiation breeding for the production of biofuel. PMID:23370702

  8. Proteomic analysis of Spirogyra varians mutant with high starch content and growth rate induced by gamma irradiation.

    PubMed

    Yoon, Minchul; Choi, Jong-il; Kim, Gwang Hoon; Kim, Dong-Ho; Park, Don-Hee

    2013-06-01

    This study was conducted to develop a high-efficiency strain of Spirogyra varians for the production of biomass by radiation breeding. The characteristics of wild-type and mutant S. varians were analyzed through phenomenological and proteomic observations. The results of our phenomenological observations of the S. varians mutant demonstrated increases in growth rate and content of chlorophyll a, b, and a + b; in particular, a significant threefold increase was observed in starch accumulation. Proteomic analysis to investigate the differences in expression between wild-type and mutant proteins identified 18 proteins with significantly different expressions. From the literature review, it was confirmed that the up-regulated proteins were mainly involved in photosynthesis, carbohydrate biosynthesis, and energy metabolism. These results suggest the possibility of algae development by radiation breeding for the production of biofuel.

  9. p53 deficiency alters the yield and spectrum of radiation-induced lacZ mutants in the brain of transgenic mice

    NASA Technical Reports Server (NTRS)

    Chang, P. Y.; Kanazawa, N.; Lutze-Mann, L.; Winegar, R. A.

    2001-01-01

    Exposure to heavy particle radiation in the galacto-cosmic environment poses a significant risk in space exploration and the evaluation of radiation-induced genetic damage in tissues, especially in the central nervous system, is an important consideration in long-term manned space missions. We used a plasmid-based transgenic mouse model system, with the pUR288 lacZ transgene integrated in the genome of every cell of C57Bl/6(lacZ) mice, to evaluate the genetic damage induced by iron particle radiation. In order to examine the importance of genetic background on the radiation sensitivity of individuals, we cross-bred p53 wild-type lacZ transgenic mice with p53 nullizygous mice, producing lacZ transgenic mice that were either hemizygous or nullizygous for the p53 tumor suppressor gene. Animals were exposed to an acute dose of 1 Gy of iron particles and the lacZ mutation frequency (MF) in the brain was measured at time intervals from 1 to 16 weeks post-irradiation. Our results suggest that iron particles induced an increase in lacZ MF (2.4-fold increase in p53+/+ mice, 1.3-fold increase in p53+/- mice and 2.1-fold increase in p53-/- mice) and that this induction is both temporally regulated and p53 genotype dependent. Characterization of mutants based on their restriction patterns showed that the majority of the mutants arising spontaneously are derived from point mutations or small deletions in all three genotypes. Radiation induced alterations in the spectrum of deletion mutants and reorganization of the genome, as evidenced by the selection of mutants containing mouse genomic DNA. These observations are unique in that mutations in brain tissue after particle radiation exposure have never before been reported owing to technical limitations in most other mutation assays.

  10. Chemical Mutagenesis of Dengue Virus Type 4 Yields Mutant Viruses Which Are Temperature Sensitive in Vero Cells or Human Liver Cells and Attenuated in Mice

    PubMed Central

    Blaney, Joseph E.; Johnson, Daniel H.; Firestone, Cai-Yen; Hanson, Christopher T.; Murphy, Brian R.; Whitehead, Stephen S.

    2001-01-01

    A recombinant live attenuated dengue virus type 4 (DEN4) vaccine candidate, 2AΔ30, was found previously to be generally well tolerated in humans, but a rash and an elevation of liver enzymes in the serum occurred in some vaccinees. 2AΔ30, a non-temperature-sensitive (non-ts) virus, contains a 30-nucleotide deletion (Δ30) in the 3′ untranslated region (UTR) of the viral genome. In the present study, chemical mutagenesis of DEN4 was utilized to generate attenuating mutations which may be useful in further attenuation of the 2AΔ30 candidate vaccine. Wild-type DEN4 2A virus was grown in Vero cells in the presence of 5-fluorouracil, and a panel of 1,248 clones were isolated. Twenty ts mutant viruses were identified that were ts in both simian Vero and human liver HuH-7 cells (n = 13) or only in HuH-7 cells (n = 7). Each of the 20 ts mutant viruses possessed an attenuation phenotype, as indicated by restricted replication in the brains of 7-day-old mice. The complete nucleotide sequence of the 20 ts mutant viruses identified nucleotide substitutions in structural and nonstructural genes as well as in the 5′ and 3′ UTRs, with more than one change occurring, in general, per mutant virus. A ts mutation in the NS3 protein (nucleotide position 4995) was introduced into a recombinant DEN4 virus possessing the Δ30 deletion, thereby creating rDEN4Δ30-4995, a recombinant virus which is ts and more attenuated than rDEN4Δ30 virus in the brains of mice. We are assembling a menu of attenuating mutations that should be useful in generating satisfactorily attenuated recombinant dengue vaccine viruses and in increasing our understanding of the pathogenesis of dengue virus. PMID:11559806

  11. Design of high-reliability low-cost amorphous silicon modules for high energy yield

    NASA Astrophysics Data System (ADS)

    Jansen, Kai W.; Varvar, Anthony; Twesme, Edward; Berens, Troy; Dhere, Neelkanth G.

    2008-08-01

    For PV modules to fulfill their intended purpose, they must generate sufficient economic return over their lifetime to justify their initial cost. Not only must modules be manufactured at a low cost/Wp with a high energy yield (kWh/kWp), they must also be designed to withstand the significant environmental stresses experienced throughout their 25+ year lifetime. Based on field experience, the most common factors affecting the lifetime energy yield of glass-based amorphous silicon (a-Si) modules have been identified; these include: 1) light-induced degradation; 2) moisture ingress and thin film corrosion; 3) transparent conductive oxide (TCO) delamination; and 4) glass breakage. The current approaches to mitigating the effect of these degradation mechanisms are discussed and the accelerated tests designed to simulate some of the field failures are described. In some cases, novel accelerated tests have been created to facilitate the development of improved manufacturing processes, including a unique test to screen for TCO delamination. Modules using the most reliable designs are tested in high voltage arrays at customer and internal test sites, as well as at independent laboratories. Data from tests at the Florida Solar Energy Center has shown that a-Si tandem modules can demonstrate an energy yield exceeding 1200 kWh/kWp/yr in a subtropical climate. In the same study, the test arrays demonstrated low long-term power loss over two years of data collection, after initial stabilization. The absolute power produced by the test arrays varied seasonally by approximately +/-7%, as expected.

  12. Quantitative insertion-site sequencing (QIseq) for high throughput phenotyping of transposon mutants

    PubMed Central

    Bronner, Iraad F.; Otto, Thomas D.; Zhang, Min; Udenze, Kenneth; Wang, Chengqi; Quail, Michael A.; Jiang, Rays H.Y.; Adams, John H.

    2016-01-01

    Genetic screening using random transposon insertions has been a powerful tool for uncovering biology in prokaryotes, where whole-genome saturating screens have been performed in multiple organisms. In eukaryotes, such screens have proven more problematic, in part because of the lack of a sensitive and robust system for identifying transposon insertion sites. We here describe quantitative insertion-site sequencing, or QIseq, which uses custom library preparation and Illumina sequencing technology and is able to identify insertion sites from both the 5′ and 3′ ends of the transposon, providing an inbuilt level of validation. The approach was developed using piggyBac mutants in the human malaria parasite Plasmodium falciparum but should be applicable to many other eukaryotic genomes. QIseq proved accurate, confirming known sites in >100 mutants, and sensitive, identifying and monitoring sites over a >10,000-fold dynamic range of sequence counts. Applying QIseq to uncloned parasites shortly after transfections revealed multiple insertions in mixed populations and suggests that >4000 independent mutants could be generated from relatively modest scales of transfection, providing a clear pathway to genome-scale screens in P. falciparum. QIseq was also used to monitor the growth of pools of previously cloned mutants and reproducibly differentiated between deleterious and neutral mutations in competitive growth. Among the mutants with fitness defects was a mutant with a piggyBac insertion immediately upstream of the kelch protein K13 gene associated with artemisinin resistance, implying mutants in this gene may have competitive fitness costs. QIseq has the potential to enable the scale-up of piggyBac-mediated genetics across multiple eukaryotic systems. PMID:27197223

  13. Water use and grain yield in drought-tolerant corn in the Texas High Plains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Drought is an important factor limiting corn (Zea mays L.) yields in the Texas High Plains, and adoption of drought-tolerant (DT) hybrids could be a management tool under water shortage. We conducted a 3-yr field study to investigate grain yield, evapotranspiration (ET), and water use efficiency (WU...

  14. Genome Signature of Artificial Selection for High Milk Yield in Holstein Cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Artificial selection for high milk yield in Holstein cattle during the past forty years achieved tremendous increases in milk yield but had an unintended consequence of reduced fertility. It was unknown how artificial selection changed the Holstein genome and what genome changes were associated wit...

  15. Compartmentalized self-replication under fast PCR cycling conditions yields Taq DNA polymerase mutants with increased DNA-binding affinity and blood resistance

    PubMed Central

    Arezi, Bahram; McKinney, Nancy; Hansen, Connie; Cayouette, Michelle; Fox, Jeffrey; Chen, Keith; Lapira, Jennifer; Hamilton, Sarah; Hogrefe, Holly

    2014-01-01

    Faster-cycling PCR formulations, protocols, and instruments have been developed to address the need for increased throughput and shorter turn-around times for PCR-based assays. Although run times can be cut by up to 50%, shorter cycle times have been correlated with lower detection sensitivity and increased variability. To address these concerns, we applied Compartmentalized Self Replication (CSR) to evolve faster-cycling mutants of Taq DNA polymerase. After five rounds of selection using progressively shorter PCR extension times, individual mutations identified in the fastest-cycling clones were randomly combined using ligation-based multi-site mutagenesis. The best-performing combinatorial mutants exhibit 35- to 90-fold higher affinity (lower Kd) for primed template and a moderate (2-fold) increase in extension rate compared to wild-type Taq. Further characterization revealed that CSR-selected mutations provide increased resistance to inhibitors, and most notably, enable direct amplification from up to 65% whole blood. We discuss the contribution of individual mutations to fast-cycling and blood-resistant phenotypes. PMID:25177317

  16. High-order harmonic generation yielding tunable extreme-ultraviolet radiation of high spectral purity.

    PubMed

    Brandi, F; Neshev, D; Ubachs, W

    2003-10-17

    Production of extreme-ultraviolet radiation by high-order harmonic generation is demonstrated to yield unprecedented spectral purity of lambda/Delta lambda=2.5 x 10(5) at wavelengths covering the entire range 40-100 nm. Tunability and sub-cm(-1) bandwidth of the harmonics are demonstrated in recordings of the He (1s4p) and Ar (3p(5)3d') resonance lines at 52.2 and 86.6 nm. Frequency shift of the harmonics due to chirp-induced phenomena are investigated and found to be small, resulting in a frequency accuracy of about 5 x 10(-7) in the domain of extreme-ultraviolet radiation.

  17. Breeding high-yielding drought-tolerant rice: genetic variations and conventional and molecular approaches

    PubMed Central

    Kumar, Arvind; Dixit, Shalabh; Ram, T.; Yadaw, R. B.; Mishra, K. K.; Mandal, N. P.

    2014-01-01

    The increased occurrence and severity of drought stress have led to a high yield decline in rice in recent years in drought-affected areas. Drought research at the International Rice Research Institute (IRRI) over the past decade has concentrated on direct selection for grain yield under drought. This approach has led to the successful development and release of 17 high-yielding drought-tolerant rice varieties in South Asia, Southeast Asia, and Africa. In addition to this, 14 quantitative trait loci (QTLs) showing a large effect against high-yielding drought-susceptible popular varieties were identified using grain yield as a selection criterion. Six of these (qDTY 1.1, qDTY 2.2, qDTY 3.1, qDTY 3.2, qDTY 6.1, and qDTY 12.1) showed an effect against two or more high-yielding genetic backgrounds in both the lowland and upland ecosystem, indicating their usefulness in increasing the grain yield of rice under drought. The yield of popular rice varieties IR64 and Vandana has been successfully improved through a well-planned marker-assisted backcross breeding approach, and QTL introgression in several other popular varieties is in progress. The identification of large-effect QTLs for grain yield under drought and the higher yield increase under drought obtained through the use of these QTLs (which has not been reported in other cereals) indicate that rice, because of its continuous cultivation in two diverse ecosystems (upland, drought tolerant, and lowland, drought susceptible), has benefited from the existence of larger genetic variability than in other cereals. This can be successfully exploited using marker-assisted breeding. PMID:25205576

  18. Breeding high-yielding drought-tolerant rice: genetic variations and conventional and molecular approaches.

    PubMed

    Kumar, Arvind; Dixit, Shalabh; Ram, T; Yadaw, R B; Mishra, K K; Mandal, N P

    2014-11-01

    The increased occurrence and severity of drought stress have led to a high yield decline in rice in recent years in drought-affected areas. Drought research at the International Rice Research Institute (IRRI) over the past decade has concentrated on direct selection for grain yield under drought. This approach has led to the successful development and release of 17 high-yielding drought-tolerant rice varieties in South Asia, Southeast Asia, and Africa. In addition to this, 14 quantitative trait loci (QTLs) showing a large effect against high-yielding drought-susceptible popular varieties were identified using grain yield as a selection criterion. Six of these (qDTY 1.1 , qDTY 2.2 , qDTY 3.1 , qDTY 3.2 , qDTY 6.1 , and qDTY 12.1 ) showed an effect against two or more high-yielding genetic backgrounds in both the lowland and upland ecosystem, indicating their usefulness in increasing the grain yield of rice under drought. The yield of popular rice varieties IR64 and Vandana has been successfully improved through a well-planned marker-assisted backcross breeding approach, and QTL introgression in several other popular varieties is in progress. The identification of large-effect QTLs for grain yield under drought and the higher yield increase under drought obtained through the use of these QTLs (which has not been reported in other cereals) indicate that rice, because of its continuous cultivation in two diverse ecosystems (upland, drought tolerant, and lowland, drought susceptible), has benefited from the existence of larger genetic variability than in other cereals. This can be successfully exploited using marker-assisted breeding.

  19. Selection and molecular characterization of a high tocopherol accumulation rice mutant line induced by gamma irradiation.

    PubMed

    Hwang, Jung Eun; Ahn, Joon-Woo; Kwon, Soon-Jae; Kim, Jin-Baek; Kim, Sang Hoon; Kang, Si-Yong; Kim, Dong Sub

    2014-11-01

    Tocopherols are micronutrients with antioxidant properties. They are synthesized by photosynthetic bacteria and plants, and play important roles in animal and human nutrition. In this study, we isolated a new rice mutant line with elevated tocopherol content (MRXII) from an in vitro mutagenized population induced by gamma irradiation. The mutant exhibited greater seed longevity than the control, indicating a crucial role for tocopherols in maintaining viability during quiescence, and displayed faster seedling growth during the early growth stage. To study the molecular mechanism underlying vitamin E biosynthesis, we examined the expression patterns of seven rice genes encoding vitamin E biosynthetic enzymes. Accumulation levels of the OsVTE2 transcript and OsVTE2 protein in the MRXII mutant were significantly higher than in the control. Sequence analysis revealed that the MRXII mutant harbored a point mutation in the OsVTE2 promoter region, which resulted in the generation of MYB transcription factor-binding cis-element. These results help identify the promoter regions that regulate OsVTE2 transcription, and offer insights into the regulation of tocopherol content.

  20. Newly identified natural high oleate mutant from Arachis hypogaea L. subsp. hypogaea

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Natural genetic variation exists in animals and plants. Mining and utilizing this variation may provide benefits for new breed/cultivar development. From screening over 4,000 cultivated peanut germplasm accessions, we identified two natural mutant lines with 80% oleic acid by gas chromatography anal...

  1. High-resolution structure of an α-spectrin SH3-domain mutant with a redesigned hydrophobic core

    PubMed Central

    Cámara-Artigas, Ana; Andújar-Sánchez, Monserrat; Ortiz-Salmerón, Emilia; Cuadri, Celia; Cobos, Eva S.; Martin-Garcia, Jose Manuel

    2010-01-01

    The α-spectrin SH3 domain (Spc-SH3) is a small modular domain which has been broadly used as a model protein in folding studies and these studies have sometimes been supported by structural information obtained from the coordinates of Spc-SH3 mutants. The structure of B5/D48G, a multiple mutant designed to improve the hydrophobic core and as a consequence the protein stability, has been solved at 1 Å resolution. The crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 24.79, b = 37.23, c = 62.95 Å. This mutant also bears a D48G substitution in the distal loop and this mutation has also been reported to increase the stability of the protein by itself. The structure of the B5/D48G mutant shows a highly packed hydrophobic core and a more ordered distal loop compared with previous Spc-SH3 structures. PMID:20823517

  2. Activation of the Saccharomyces cerevisiae filamentation/invasion pathway by osmotic stress in high-osmolarity glycogen pathway mutants.

    PubMed

    Davenport, K D; Williams, K E; Ullmann, B D; Gustin, M C

    1999-11-01

    Mitogen-activated protein kinase (MAPK) cascades are frequently used signal transduction mechanisms in eukaryotes. Of the five MAPK cascades in Saccharomyces cerevisiae, the high-osmolarity glycerol response (HOG) pathway functions to sense and respond to hypertonic stress. We utilized a partial loss-of-function mutant in the HOG pathway, pbs2-3, in a high-copy suppressor screen to identify proteins that modulate growth on high-osmolarity media. Three high-copy suppressors of pbs2-3 osmosensitivity were identified: MSG5, CAK1, and TRX1. Msg5p is a dual-specificity phosphatase that was previously demonstrated to dephosphorylate MAPKs in yeast. Deletions of the putative MAPK targets of Msg5p revealed that kss1delta could suppress the osmosensitivity of pbs2-3. Kss1p is phosphorylated in response to hyperosmotic shock in a pbs2-3 strain, but not in a wild-type strain nor in a pbs2-3 strain overexpressing MSG5. Both TEC1 and FRE::lacZ expressions are activated in strains lacking a functional HOG pathway during osmotic stress in a filamentation/invasion-pathway-dependent manner. Additionally, the cellular projections formed by a pbs2-3 mutant on high osmolarity are absent in strains lacking KSS1 or STE7. These data suggest that the loss of filamentation/invasion pathway repression contributes to the HOG mutant phenotype.

  3. A quantitative method for defining high-arched palate using the Tcof1(+/-) mutant mouse as a model.

    PubMed

    Conley, Zachary R; Hague, Molly; Kurosaka, Hiroshi; Dixon, Jill; Dixon, Michael J; Trainor, Paul A

    2016-07-15

    The palate functions as the roof of the mouth in mammals, separating the oral and nasal cavities. Its complex embryonic development and assembly poses unique susceptibilities to intrinsic and extrinsic disruptions. Such disruptions may cause failure of the developing palatal shelves to fuse along the midline resulting in a cleft. In other cases the palate may fuse at an arch, resulting in a vaulted oral cavity, termed high-arched palate. There are many models available for studying the pathogenesis of cleft palate but a relative paucity for high-arched palate. One condition exhibiting either cleft palate or high-arched palate is Treacher Collins syndrome, a congenital disorder characterized by numerous craniofacial anomalies. We quantitatively analyzed palatal perturbations in the Tcof1(+/-) mouse model of Treacher Collins syndrome, which phenocopies the condition in humans. We discovered that 46% of Tcof1(+/-) mutant embryos and new born pups exhibit either soft clefts or full clefts. In addition, 17% of Tcof1(+/-) mutants were found to exhibit high-arched palate, defined as two sigma above the corresponding wild-type population mean for height and angular based arch measurements. Furthermore, palatal shelf length and shelf width were decreased in all Tcof1(+/-) mutant embryos and pups compared to controls. Interestingly, these phenotypes were subsequently ameliorated through genetic inhibition of p53. The results of our study therefore provide a simple, reproducible and quantitative method for investigating models of high-arched palate.

  4. Activation of the Saccharomyces cerevisiae filamentation/invasion pathway by osmotic stress in high-osmolarity glycogen pathway mutants

    NASA Technical Reports Server (NTRS)

    Davenport, K. D.; Williams, K. E.; Ullmann, B. D.; Gustin, M. C.; McIntire, L. V. (Principal Investigator)

    1999-01-01

    Mitogen-activated protein kinase (MAPK) cascades are frequently used signal transduction mechanisms in eukaryotes. Of the five MAPK cascades in Saccharomyces cerevisiae, the high-osmolarity glycerol response (HOG) pathway functions to sense and respond to hypertonic stress. We utilized a partial loss-of-function mutant in the HOG pathway, pbs2-3, in a high-copy suppressor screen to identify proteins that modulate growth on high-osmolarity media. Three high-copy suppressors of pbs2-3 osmosensitivity were identified: MSG5, CAK1, and TRX1. Msg5p is a dual-specificity phosphatase that was previously demonstrated to dephosphorylate MAPKs in yeast. Deletions of the putative MAPK targets of Msg5p revealed that kss1delta could suppress the osmosensitivity of pbs2-3. Kss1p is phosphorylated in response to hyperosmotic shock in a pbs2-3 strain, but not in a wild-type strain nor in a pbs2-3 strain overexpressing MSG5. Both TEC1 and FRE::lacZ expressions are activated in strains lacking a functional HOG pathway during osmotic stress in a filamentation/invasion-pathway-dependent manner. Additionally, the cellular projections formed by a pbs2-3 mutant on high osmolarity are absent in strains lacking KSS1 or STE7. These data suggest that the loss of filamentation/invasion pathway repression contributes to the HOG mutant phenotype.

  5. A cytochrome c mutant with high electron transfer and antioxidant activities but devoid of apoptogenic effect.

    PubMed Central

    Abdullaev, Ziedulla Kh; Bodrova, Marina E; Chernyak, Boris V; Dolgikh, Dmitry A; Kluck, Ruth M; Pereverzev, Mikhail O; Arseniev, Alexander S; Efremov, Roman G; Kirpichnikov, Mikhail P; Mokhova, Elena N; Newmeyer, Donald D; Roder, Heinrich; Skulachev, Vladimir P

    2002-01-01

    A cytochrome c mutant lacking apoptogenic function but competent in electron transfer and antioxidant activities has been constructed. To this end, mutant species of horse and yeast cytochromes c with substitutions in the N-terminal alpha-helix or position 72 were obtained. It was found that yeast cytochrome c was much less effective than the horse protein in activating respiration of rat liver mitoplasts deficient in endogenous cytochrome c as well as in inhibition of H(2)O(2) production by the initial segment of the respiratory chain of intact rat heart mitochondria. The major role in the difference between the horse and yeast proteins was shown to be played by the amino acid residue in position 4 (glutamate in horse, and lysine in yeast; horse protein numbering). A mutant of the yeast cytochrome c containing K4E and some other "horse" modifications in the N-terminal alpha-helix, proved to be (i) much more active in electron transfer and antioxidant activity than the wild-type yeast cytochrome c and (ii), like the yeast cytochrome c, inactive in caspase stimulation, even if added in 400-fold excess compared with the horse protein. Thus this mutant seems to be a good candidate for knock-in studies of the role of cytochrome c-mediated apoptosis, in contrast with the horse K72R, K72G, K72L and K72A mutant cytochromes that at low concentrations were less active in apoptosis than the wild-type, but were quite active when the concentrations were increased by a factor of 2-12. PMID:11879204

  6. Neglecting rice milling yield and quality underestimates economic losses from high-temperature stress.

    PubMed

    Lyman, Nathaniel B; Jagadish, Krishna S V; Nalley, L Lanier; Dixon, Bruce L; Siebenmorgen, Terry

    2013-01-01

    Future increases in global surface temperature threaten those worldwide who depend on rice production for their livelihoods and food security. Past analyses of high-temperature stress on rice production have focused on paddy yield and have failed to account for the detrimental impact of high temperatures on milling quality outcomes, which ultimately determine edible (marketable) rice yield and market value. Using genotype specific rice yield and milling quality data on six common rice varieties from Arkansas, USA, combined with on-site, half-hourly and daily temperature observations, we show a nonlinear effect of high-temperature stress exposure on yield and milling quality. A 1 °C increase in average growing season temperature reduces paddy yield by 6.2%, total milled rice yield by 7.1% to 8.0%, head rice yield by 9.0% to 13.8%, and total milling revenue by 8.1% to 11.0%, across genotypes. Our results indicate that failure to account for changes in milling quality leads to understatement of the impacts of high temperatures on rice production outcomes. These dramatic losses result from reduced paddy yield and increased percentages of chalky and broken kernels, which together decrease the quantity and market value of milled rice. Recently published estimates show paddy yield reductions of up to 10% across the major rice-producing regions of South and Southeast Asia due to rising temperatures. The results of our study suggest that the often-cited 10% figure underestimates the economic implications of climate change for rice producers, thus potentially threatening future food security for global rice producers and consumers.

  7. Neglecting Rice Milling Yield and Quality Underestimates Economic Losses from High-Temperature Stress

    PubMed Central

    Lyman, Nathaniel B.; Jagadish, Krishna S. V.; Nalley, L. Lanier; Dixon, Bruce L.; Siebenmorgen, Terry

    2013-01-01

    Future increases in global surface temperature threaten those worldwide who depend on rice production for their livelihoods and food security. Past analyses of high-temperature stress on rice production have focused on paddy yield and have failed to account for the detrimental impact of high temperatures on milling quality outcomes, which ultimately determine edible (marketable) rice yield and market value. Using genotype specific rice yield and milling quality data on six common rice varieties from Arkansas, USA, combined with on-site, half-hourly and daily temperature observations, we show a nonlinear effect of high-temperature stress exposure on yield and milling quality. A 1°C increase in average growing season temperature reduces paddy yield by 6.2%, total milled rice yield by 7.1% to 8.0%, head rice yield by 9.0% to 13.8%, and total milling revenue by 8.1% to 11.0%, across genotypes. Our results indicate that failure to account for changes in milling quality leads to understatement of the impacts of high temperatures on rice production outcomes. These dramatic losses result from reduced paddy yield and increased percentages of chalky and broken kernels, which together decrease the quantity and market value of milled rice. Recently published estimates show paddy yield reductions of up to 10% across the major rice-producing regions of South and Southeast Asia due to rising temperatures. The results of our study suggest that the often-cited 10% figure underestimates the economic implications of climate change for rice producers, thus potentially threatening future food security for global rice producers and consumers. PMID:23991056

  8. High-yield synthesis of bioactive ethyl cinnamate by enzymatic esterification of cinnamic acid.

    PubMed

    Wang, Yun; Zhang, Dong-Hao; Zhang, Jiang-Yan; Chen, Na; Zhi, Gao-Ying

    2016-01-01

    In this paper, Lipozyme TLIM-catalyzed synthesis of ethyl cinnamate through esterification of cinnamic acid with ethanol was studied. In order to increase the yield of ethyl cinnamate, several media, including acetone, isooctane, DMSO and solvent-free medium, were investigated in this reaction. The reaction showed a high yield by using isooctane as reaction medium, which was found to be much higher than the yields reported previously. Furthermore, several parameters such as shaking rate, water activity, reaction temperature, substrate molar ratio and enzyme loading had important influences on this reaction. For instance, when temperature increased from 10 to 50 °C, the initial reaction rate increased by 18 times and the yield of ethyl cinnamate increased by 6.2 times. Under the optimum conditions, lipase-catalyzed synthesis of ethyl cinnamate gave a maximum yield of 99%, which was of general interest for developing industrial processes for the preparation of ethyl cinnamate.

  9. Fatty acids, calcium soaps of fatty acids, and cottonseeds fed to high yielding cows.

    PubMed

    Sklan, D; Ashkenazi, R; Braun, A; Devorin, A; Tabori, K

    1992-09-01

    We examined the effects of dietary fat as cottonseed, fatty acids, or calcium soaps of fatty acids in the rations of high yielding lactating cows receiving low forage. Experiments were with isoenergetic, isonitrogenous diets containing equal amounts of forage. Inclusion of up to 510 g/d of fatty acids in the ration enhanced FCM yield. With cottonseed, increased FCM was mainly due to increased fat yield. Dietary fatty acids tended to increase milk in mid and late lactation and to decrease fat percentage. Calcium soaps of fatty acids enhanced FCM, particularly in early lactation. Feeding cottonseed and fatty acids together did not enhance yield. Effects described may be attributed in part to changes in ruminal fermentation in which cottonseed increased acetate concentrations and fatty acids decreased the ratio of acetate and butyrate to propionate and in part to enhanced efficiency of milk yield when fat was included in the ration.

  10. Ultra fast elemental synthesis of high yield copper Chevrel phase with high electrochemical performance

    SciTech Connect

    Gershinsky, Gregory; Haik, Ortal; Salitra, Gregory; Grinblat, Judith; Levi, Elena; Daniel Nessim, Gilbert; Zinigrad, Ella; Aurbach, Doron

    2012-04-15

    Self-propagating High-temperature Synthesis (SHS) was applied for the first time to prepare Chevrel phases, M{sub x}Mo{sub 6}T{sub 8} (M=metal, T=S, Se). Combined electron microscopy and X-ray powder diffraction were used to clarify the chemical reactions in the Cu-Mo-S system. It was shown that the replacement of the frontal combustion by thermal explosion increased the Cu{sub 2}Mo{sub 6}S{sub 8} yield from 86 to 96%, while the synthesis remained ultra-fast: 10-20 min in a hot furnace (1000 Degree-Sign C), as compared to at least 17 h of heating for the conventional solid state technique. The synthesized material conformed to the requirements of cathode precursors for Mg batteries, and its electrochemically activity was similar to that of the conventional product. - Graphical abstarct: Schematic diagram of the combustion process. Highlights: Black-Right-Pointing-Pointer Self-propagating high-temperature synthesis was applied for the first time for preparing Chevrel phases. Black-Right-Pointing-Pointer Combined electron microscopy and X-ray powder diffraction were used to clarify the chemical reactions in the Cu-Mo-S system. Black-Right-Pointing-Pointer The synthesized Cu{sub 2}Mo{sub 6}S{sub 8} product conforms to the requirements of cathode precursors for Mg batteries.

  11. Effect of high oleic acid soybean on seed oil, protein concentration, and yield

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Soybeans with high oleic acid content are desired by oil processors because of their improved oxidative stability for broader use in food, fuel and other products. However, non-GMO high-oleic soybeans have tended to have low seed yield. The objective of this study was to test non-GMO, high-oleic s...

  12. The diageotropica mutant of tomato lacks high specific activity auxin binding sites

    NASA Technical Reports Server (NTRS)

    Hicks, G. R.; Rayle, D. L.; Lomax, T. L.

    1989-01-01

    Tomato plants homozygous for the diageotropica (dgt) mutation exhibit morphological and physiological abnormalities which suggest that they are unable to respond to the plant growth hormone auxin (indole-3-acetic acid). The photoaffinity auxin analog [3H]5N3-IAA specifically labels a polypeptide doublet of 40 and 42 kilodaltons in membrane preparations from stems of the parental variety, VFN8, but not from stems of plants containing the dgt mutation. In roots of the mutant plants, however, labeling is indistinguishable from that in VFN8. These data suggest that the two polypeptides are part of a physiologically important auxin receptor system, which is altered in a tissue-specific manner in the mutant.

  13. High-throughput quantitative analysis with cell growth kinetic curves for low copy number mutant cells.

    PubMed

    Xing, James Z; Gabos, Stephan; Huang, Biao; Pan, Tianhong; Huang, Min; Chen, Jie

    2012-10-01

    The mutation rate in cells induced by environmental genotoxic hazards is very low and difficult to detect using traditional cell counting assays. The established genetic toxicity tests currently recognized by regulatory authorities, such as conventional Ames and hypoxanthine guanine phosphoribosyl-transferase (HPRT) assays, are not well suited for higher-throughput screening as they require large amounts of test compounds and are very time consuming. In this study, we developed a novel cell-based assay for quantitative analysis of low numbers of cell copies with HPRT mutation induced by an environmental mutagen. The HPRT gene mutant cells induced by the mutagen were selected by 6-thioguanine (6-TG) and the cell's kinetic growth curve monitored by a real-time cell electronic sensor (RT-CES) system. When a threshold is set at a certain cell index (CI) level, samples with different initial mutant cell copies take different amounts of time in order for their growth (or CI accumulation) to cross this threshold. The more cells that are initially seeded in the test well, the faster the cell accumulation and therefore the shorter the time required to cross this threshold. Therefore, the culture time period required to cross the threshold of each sample corresponds to the original number of cells in the sample. A mutant cell growth time threshold (MT) value of each sample can be calculated to predict the number of original mutant cells. For mutagenesis determination, the RT-CES assay displayed an equal sensitivity (p > 0.05) and coefficients of variation values with good correlation to conventional HPRT mutagenic assays. Most importantly, the RT-CES mutation assay has a higher throughput than conventional cellular assays.

  14. High-photon-yield scintillation detector with Ar/CF4 and glass gas electron multiplier

    NASA Astrophysics Data System (ADS)

    Fujiwara, Takeshi; Mitsuya, Yuki; Yanagida, Takayuki; Saito, Takumi; Toyokawa, Hiroyuki; Takahashi, Hiroyuki

    2016-10-01

    The glass made gas electron multiplier (GEM) and Ar/CF4-gas-based gaseous detector is developed as a scintillation detector and ultra high photon yield is demonstrated. The light yield of a glass GEM (G-GEM)-based gaseous detector is estimated to be 85,000 photons/keV, which is three orders of magnitude brighter than inorganic scintillators. The radioluminescence spectrum peak appeared at around 620 nm, which matches the spectral response of commonly used photosensors such as photomultiplier tubes, photodiodes, CMOSs, CCDs, and other photo-sensors. In X-ray spectroscopy, the light yield showed excellent proportionality and the device was successfully operated as a gas proportional scintillation counter. With this design, we obtained a high photon yield of the G-GEM, which has the further advantage of being much more sensitive to low-energy radiation than solid-scintillator-based detectors.

  15. High levels of plasma cortisol and impaired hypoosmoregulation in a mutant medaka deficient in P450c17I.

    PubMed

    Takahashi, Hideya; Sato, Tadashi; Ikeuchi, Toshitaka; Saito, Kazuhiro; Sakaizumi, Mitsuru; Sakamoto, Tatsuya

    2016-07-15

    scl is a spontaneous medaka mutant deficient in P450c17I, which is required for production of sex steroids, but not of cortisol, the major role of which is osmoregulation in teleost fish. The scl mutant provides a new model to study the functions of these hormones. We first found that fish homozygous for this mutation have plasma cortisol constitutively at a high physiological level (1000 nM). Since we previously showed that this level reversed the seawater-type differentiation of the medaka gastrointestinal tract, hypoosmoregulation of the scl mutant was analyzed. Muscle water contents in freshwater were normal in scl homozygotes, but the contents were lower than those of the wild type (WT) after seawater transfer. There were no differences in gill mRNA levels of corticosteroid receptors or ion transporters between scl homozygotes and WT. In the intestine, expression of glucocorticoid receptors and Na(+)/K(+)/2Cl(-) cotransporter were induced in WT during seawater acclimation, but not in scl homozygotes. The high plasma cortisol may prevent hypoosmoregulation by inhibition of increased intestinal water absorption, essentially by the Na(+)/K(+)/2Cl(-) cotransporter, in seawater. PMID:27107936

  16. High Throughput Sequencing Identifies Misregulated Genes in the Drosophila Polypyrimidine Tract-Binding Protein (hephaestus) Mutant Defective in Spermatogenesis.

    PubMed

    Sridharan, Vinod; Heimiller, Joseph; Robida, Mark D; Singh, Ravinder

    2016-01-01

    The Drosophila polypyrimidine tract-binding protein (dmPTB or hephaestus) plays an important role during spermatogenesis. The heph2 mutation in this gene results in a specific defect in spermatogenesis, causing aberrant spermatid individualization and male sterility. However, the array of molecular defects in the mutant remains uncharacterized. Using an unbiased high throughput sequencing approach, we have identified transcripts that are misregulated in this mutant. Aberrant transcripts show altered expression levels, exon skipping, and alternative 5' ends. We independently verified these findings by reverse-transcription and polymerase chain reaction (RT-PCR) analysis. Our analysis shows misregulation of transcripts that have been connected to spermatogenesis, including components of the actomyosin cytoskeletal apparatus. We show, for example, that the Myosin light chain 1 (Mlc1) transcript is aberrantly spliced. Furthermore, bioinformatics analysis reveals that Mlc1 contains a high affinity binding site(s) for dmPTB and that the site is conserved in many Drosophila species. We discuss that Mlc1 and other components of the actomyosin cytoskeletal apparatus offer important molecular links between the loss of dmPTB function and the observed developmental defect in spermatogenesis. This study provides the first comprehensive list of genes misregulated in vivo in the heph2 mutant in Drosophila and offers insight into the role of dmPTB during spermatogenesis.

  17. High Throughput Sequencing Identifies Misregulated Genes in the Drosophila Polypyrimidine Tract-Binding Protein (hephaestus) Mutant Defective in Spermatogenesis

    PubMed Central

    Sridharan, Vinod; Heimiller, Joseph; Robida, Mark D.; Singh, Ravinder

    2016-01-01

    The Drosophila polypyrimidine tract-binding protein (dmPTB or hephaestus) plays an important role during spermatogenesis. The heph2 mutation in this gene results in a specific defect in spermatogenesis, causing aberrant spermatid individualization and male sterility. However, the array of molecular defects in the mutant remains uncharacterized. Using an unbiased high throughput sequencing approach, we have identified transcripts that are misregulated in this mutant. Aberrant transcripts show altered expression levels, exon skipping, and alternative 5’ ends. We independently verified these findings by reverse-transcription and polymerase chain reaction (RT-PCR) analysis. Our analysis shows misregulation of transcripts that have been connected to spermatogenesis, including components of the actomyosin cytoskeletal apparatus. We show, for example, that the Myosin light chain 1 (Mlc1) transcript is aberrantly spliced. Furthermore, bioinformatics analysis reveals that Mlc1 contains a high affinity binding site(s) for dmPTB and that the site is conserved in many Drosophila species. We discuss that Mlc1 and other components of the actomyosin cytoskeletal apparatus offer important molecular links between the loss of dmPTB function and the observed developmental defect in spermatogenesis. This study provides the first comprehensive list of genes misregulated in vivo in the heph2 mutant in Drosophila and offers insight into the role of dmPTB during spermatogenesis. PMID:26942929

  18. Improvement of L-lactic acid production by osmotic-tolerant mutant of Lactobacillus casei at high temperature.

    PubMed

    Ge, Xiang-Yang; Yuan, Jian; Qin, Hao; Zhang, Wei-Guo

    2011-01-01

    L-Lactic acid production by Lactobacillus casei was used as a model to study the mechanism of substrate inhibition and the strategy for enhancing L-lactic acid production. It was found that the concentration of cell growth and L-lactate decreased with the increase of glucose concentration and fermentation temperature. To enhance the osmotic stress resistance of the strain at high temperature, a mutant G-03 was screened and selected with 360 g/L glucose at 45°C as the selective criterion. To further increase the cell growth for lactic acid production, 3 g/L of biotin was supplemented to the medium. As a result, L: -lactate concentration by the mutant G-03 reached 198.2 g/L (productivity of 5.5 g L(-1) h(-1)) at 41°C in a 7-L fermentor with 210 g/L glucose as carbon source. L: -Lactate concentration and productivity of mutant G-03 were 115.2% and 97.8% higher than those of the parent strain, respectively. The strategy for enhancing L: -lactic acid production by increasing osmotic stress resistance at high temperature may provide an alternative approach to enhance organic acid production with other strains. PMID:20857288

  19. High Yield Synthesis of Aspect Ratio Controlled Graphenic Materials from Anthracite Coal in Supercritical Fluids.

    PubMed

    Sasikala, Suchithra Padmajan; Henry, Lucile; Yesilbag Tonga, Gulen; Huang, Kai; Das, Riddha; Giroire, Baptiste; Marre, Samuel; Rotello, Vincent M; Penicaud, Alain; Poulin, Philippe; Aymonier, Cyril

    2016-05-24

    This paper rationalizes the green and scalable synthesis of graphenic materials of different aspect ratios using anthracite coal as a single source material under different supercritical environments. Single layer, monodisperse graphene oxide quantum dots (GQDs) are obtained at high yield (55 wt %) from anthracite coal in supercritical water. The obtained GQDs are ∼3 nm in lateral size and display a high fluorescence quantum yield of 28%. They show high cell viability and are readily used for imaging cancer cells. In an analogous experiment, high aspect ratio graphenic materials with ribbon-like morphology (GRs) are synthesized from the same source material in supercritical ethanol at a yield of 6.4 wt %. A thin film of GRs with 68% transparency shows a surface resistance of 9.3 kΩ/sq. This is apparently the demonstration of anthracite coal as a source for electrically conductive graphenic materials. PMID:27135862

  20. Specific interaction of mutant p53 with regions of matrix attachment region DNA elements (MARs) with a high potential for base-unpairing

    PubMed Central

    Will, Katrin; Warnecke, Gabriele; Wiesmüller, Lisa; Deppert, Wolfgang

    1998-01-01

    Mutant, but not wild-type p53 binds with high affinity to a variety of MAR-DNA elements (MARs), suggesting that MAR-binding of mutant p53 relates to the dominant-oncogenic activities proposed for mutant p53. MARs recognized by mutant p53 share AT richness and contain variations of an AATATATTT “DNA-unwinding motif,” which enhances the structural dynamics of chromatin and promotes regional DNA base-unpairing. Mutant p53 specifically interacted with MAR-derived oligonucleotides carrying such unwinding motifs, catalyzing DNA strand separation when this motif was located within a structurally labile sequence environment. Addition of GC-clamps to the respective MAR-oligonucleotides or introducing mutations into the unwinding motif strongly reduced DNA strand separation, but supported the formation of tight complexes between mutant p53 and such oligonucleotides. We conclude that the specific interaction of mutant p53 with regions of MAR-DNA with a high potential for base-unpairing provides the basis for the high-affinity binding of mutant p53 to MAR-DNA. PMID:9811860

  1. High throughput gene complementation screening permits identification of a mammalian mitochondrial protein synthesis (ρ(-)) mutant.

    PubMed

    Potluri, Prasanth; Procaccio, Vincent; Scheffler, Immo E; Wallace, Douglas C

    2016-08-01

    To identify nuclear DNA (nDNA) oxidative phosphorylation (OXPHOS) gene mutations using cultured cells, we have developed a complementation system based on retroviral transduction with a full length cDNA expression library and selection for OXHOS function by growth in galactose. We have used this system to transduce the Chinese hamster V79-G7 OXPHOS mutant cell line with a defect in mitochondrial protein synthesis. The complemented cells were found to have acquired the cDNA for the bS6m polypeptide of the small subunit of the mitochondrial ribosome. bS6m is a 14 kDa polypeptide located on the outside of the mitochondrial 28S ribosomal subunit and interacts with the rRNA. The V79-G7 mutant protein was found to harbor a methionine to threonine missense mutation at codon 13. The hamster bS6m null mutant could also be complemented by its orthologs from either mouse or human. bS6m protein tagged at its C-terminus by HA, His or GFP localized to the mitochondrion and was fully functional. Through site-directed mutagenesis we identified the probable RNA interacting residues of the bS6m peptide and tested the functional significance of mammalian specific C-terminal region. The N-terminus of the bS6m polypeptide functionally corresponds to that of the prokaryotic small ribosomal subunit, but deletion of C-terminal residues along with the zinc ion coordinating cysteine had no functional effect. Since mitochondrial diseases can result from hundreds to thousands of different nDNA gene mutations, this one step viral complementation cloning may facilitate the molecular diagnosis of a range of nDNA mitochondrial disease mutations. This article is part of a Special Issue entitled 'EBEC 2016: 19th European Bioenergetics Conference, Riva del Garda, Italy, July 2-6, 2016', edited by Prof. Paolo Bernardi. PMID:26946086

  2. Creating high yield water soluble luminescent graphene quantum dots via exfoliating and disintegrating carbon nanotubes and graphite flakes.

    PubMed

    Lin, Liangxu; Zhang, Shaowei

    2012-10-21

    We have developed an effective method to exfoliate and disintegrate multi-walled carbon nanotubes and graphite flakes. With this technique, high yield production of luminescent graphene quantum dots with high quantum yield and low oxidization can be achieved.

  3. Kinetic Characterization of High-Activity Mutants of Human Butyrylcholinesterase for Cocaine Metabolite Norcocaine

    PubMed Central

    Zhan, Max; Hou, Shurong; Zhan, Chang-Guo; Zheng, Fang

    2015-01-01

    It has been known that cocaine produces the toxic and physiological effects through not only cocaine itself but also norcocaine formed from cocaine oxidation catalyzed by microsomal cytochrome P450 3A4 in the human liver. The catalytic parameters (kcat and KM) of human butyrylcholinesterase (BChE) and its three mutants (i.e. the A199S/S287G/A328W/Y332G, A199S/F227A/S287G/A328W/E441D, and A199S/F227A/S287G/A328W/Y332G mutants) for norcocaine have been characterized in the present study, for the first time, in comparison with those for cocaine. Based on the obtained kinetic data, wild-type human BChE has a significantly lower catalytic activity for norcocaine (kcat = 2.8 min−1, KM = 15 μM, and kcat/KM = 1.87 × 105 M−1 min−1) compared to its catalytic activity for (−)-cocaine. The BChE mutants examined in this study have considerably improved catalytic activities against both cocaine and norcocaine compared to the wild-type enzyme. Within the enzymes examined in this study, the A199S/F227A/S287G/A328W/Y332G mutant (CocH3) is identified as the most efficient enzyme for hydrolyzing both cocaine and norcocaine. CocH3 has a 1080-fold improved catalytic efficiency for norcocaine (kcat = 2610 min−1, KM = 13 μM, and kcat/KM = 2.01 × 108 M−1 min−1) and a 2020-fold improved catalytic efficiency for cocaine. It has been demonstrated that CocH3 as an exogenous enzyme can rapidly metabolize norcocaine, in addition to cocaine, in rats. Further kinetic modeling has suggested that CocH3 with an identical concentration as that of the endogenous BChE in human plasma can effectively eliminate both cocaine and norcocaine in a simplified kinetic model of cocaine abuse. PMID:24125115

  4. Spirogyra varians mutant generated by high dose gamma-irradiation shows increased antioxidant properties

    NASA Astrophysics Data System (ADS)

    Lee, Hak-Jyung; Yoon, Minchul; Sung, Nak-Yun; Choi, Jong-il

    2012-08-01

    The aim of this study was to evaluate the antioxidant properties of a Spirogyra varians mutant (Mut) produced by gamma irradiation. Methanol extracts were prepared from Spirogyra varians wild-type and Mut plants, and their antioxidant activities and total phenolic content (TPC) were determined. Antioxidant parameters, including the 2-diphenyl-1-picrylhydrazyl radical-scavenging activity and ferric-reducing/antioxidant power, were higher in the Mut extract. Moreover, the TPC level was higher (P<0.05) in the Mut methanol extract. Therefore, these results suggest that gamma irradiation-induced S. varians Mut has superior antioxidant properties.

  5. High vapor pressure deficit drives salt-stress-induced rice yield losses in India.

    PubMed

    Tack, Jesse; Singh, Rakesh K; Nalley, Lawton L; Viraktamath, Basavaraj C; Krishnamurthy, Saraswathipura L; Lyman, Nate; Jagadish, Krishna S V

    2015-04-01

    Flooded rice is grown across wide geographic boundaries from as far north as Manchuria and as far south as Uruguay and New South Wales, primarily because of its adaptability across diverse agronomic and climatic conditions. Salt-stress damage, a common occurrence in delta and coastal rice production zones, could be heightened by the interactions between high temperature and relative humidity (vapor pressure deficit--VPD). Using temporal and spatial observations spanning 107 seasons and 19 rice-growing locations throughout India with varying electrical conductivity (EC), including coastal saline, inland saline, and alkaline soils, we quantified the proportion of VPD inducing salinity damage in rice. While controlling for time-invariant factors such as trial locations, rice cultivars, and soil types, our regression analysis indicates that EC has a nonlinear detrimental effect on paddy rice yield. Our estimates suggest these yield reductions become larger at higher VPD. A one standard deviation (SD) increase in EC from its mean value is associated with 1.68% and 4.13% yield reductions at median and maximum observed VPD levels, respectively. Yield reductions increase roughly sixfold when the one SD increase is taken from the 75th percentile of EC. In combination, high EC and VPD generate near catastrophic crop loss as predicted yield approaches zero. If higher VPD levels driven by global warming materialize in conjunction with rising sea levels or salinity incursion in groundwater, this interaction becomes an important and necessary predictor of expected yield losses and global food security. PMID:25379616

  6. High vapor pressure deficit drives salt-stress-induced rice yield losses in India.

    PubMed

    Tack, Jesse; Singh, Rakesh K; Nalley, Lawton L; Viraktamath, Basavaraj C; Krishnamurthy, Saraswathipura L; Lyman, Nate; Jagadish, Krishna S V

    2015-04-01

    Flooded rice is grown across wide geographic boundaries from as far north as Manchuria and as far south as Uruguay and New South Wales, primarily because of its adaptability across diverse agronomic and climatic conditions. Salt-stress damage, a common occurrence in delta and coastal rice production zones, could be heightened by the interactions between high temperature and relative humidity (vapor pressure deficit--VPD). Using temporal and spatial observations spanning 107 seasons and 19 rice-growing locations throughout India with varying electrical conductivity (EC), including coastal saline, inland saline, and alkaline soils, we quantified the proportion of VPD inducing salinity damage in rice. While controlling for time-invariant factors such as trial locations, rice cultivars, and soil types, our regression analysis indicates that EC has a nonlinear detrimental effect on paddy rice yield. Our estimates suggest these yield reductions become larger at higher VPD. A one standard deviation (SD) increase in EC from its mean value is associated with 1.68% and 4.13% yield reductions at median and maximum observed VPD levels, respectively. Yield reductions increase roughly sixfold when the one SD increase is taken from the 75th percentile of EC. In combination, high EC and VPD generate near catastrophic crop loss as predicted yield approaches zero. If higher VPD levels driven by global warming materialize in conjunction with rising sea levels or salinity incursion in groundwater, this interaction becomes an important and necessary predictor of expected yield losses and global food security.

  7. Quantitative digital image analysis of chromogenic assays for high throughput screening of alpha-amylase mutant libraries.

    PubMed

    Shankar, Manoharan; Priyadharshini, Ramachandran; Gunasekaran, Paramasamy

    2009-08-01

    An image analysis-based method for high throughput screening of an alpha-amylase mutant library using chromogenic assays was developed. Assays were performed in microplates and high resolution images of the assay plates were read using the Virtual Microplate Reader (VMR) script to quantify the concentration of the chromogen. This method is fast and sensitive in quantifying 0.025-0.3 mg starch/ml as well as 0.05-0.75 mg glucose/ml. It was also an effective screening method for improved alpha-amylase activity with a coefficient of variance of 18%.

  8. The syntheiss of high yields of full-length reverse transcripts of globin mRNA.

    PubMed Central

    Friedman, E Y; Rosbash, M

    1977-01-01

    Conditions have been determined under which reverse transcriptase catalyzes the synthesis of the high yields of full length complementary deoxyribonucleic acid (cDNA). These conditions depend not only on the cencentration of deoxynucleoside triphosphates (1) but also on the concentration of reverse transcriptase. An analysis of the kinetics of cDNA synthesis and the size of cDNA synthesized as a function of time under different conditions indicates that the mechanism of action of reverse transcriptase is partially distributive. This accounts for the necessity of a high enzyme concentration to obtain high yields of full length cDNA. Additional experiments indicate that the yield of cDNA is limited by the fact that the template mRNA is rapidly inactivated. This is most likely due to the fact that the product cDNA is hydrogen bonded to the template mRNA during synthesis. Images PMID:73163

  9. Barley mutants with low rates of endosperm starch synthesis have low grain dormancy and high susceptibility to preharvest sprouting.

    PubMed

    Howard, Thomas P; Fahy, Brendan; Craggs, Alice; Mumford, Rachel; Leigh, Fiona; Howell, Phil; Greenland, Andy; Smith, Alison M

    2012-04-01

    • Studies of embryo dormancy in relation to preharvest sprouting (PHS) in cereals have focused on ABA and other hormones. The relationship between these phenomena and the rate of grain filling has not been investigated. • A collection of barley mutants impaired in starch synthesis was assessed for preharvest sprouting in the field. In subsequent glasshouse experiments, developing grains were assayed for germination index, sugars, abscisic acid (ABA) and the effects of temperature and exogenous ABA on germination. • Mutant lines displayed greater preharvest sprouting in the field than parental lines. In the glasshouse, nondeep physiological dormancy was reduced in developing grains of five lines with mutations affecting proteins involved in endosperm starch synthesis. Inhibition of germination by exogenous ABA and elevated temperature was decreased in developing mutant grains. Sugar concentrations were high but embryo and endosperm ABA contents were unaltered. • We reveal a direct connection between grain filling and the extent of grain dormancy. Impaired endosperm starch synthesis directly influences the acquisition of embryo dormancy, perhaps because endosperm sugar concentrations modulate the ABA responsiveness of the embryo. Thus environmental or genetic factors that reduce grain filling are likely to reduce dormancy and enhance susceptibility to PHS. PMID:22300545

  10. Use of a highly transparent zebrafish mutant for investigations in the development of the vertebrate auditory system (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Wisniowiecki, Anna M.; Mattison, Scott P.; Kim, Sangmin; Riley, Bruce; Applegate, Brian E.

    2016-03-01

    Zebrafish, an auditory specialist among fish, offer analogous auditory structures to vertebrates and is a model for hearing and deafness in vertebrates, including humans. Nevertheless, many questions remain on the basic mechanics of the auditory pathway. Phase-sensitive Optical Coherence Tomography has been proven as valuable technique for functional vibrometric measurements in the murine ear. Such measurements are key to building a complete understanding of auditory mechanics. The application of such techniques in the zebrafish is impeded by the high level of pigmentation, which develops superior to the transverse plane and envelops the auditory system superficially. A zebrafish double mutant for nacre and roy (mitfa-/- ;roya-/- [casper]), which exhibits defects for neural-crest derived melanocytes and iridophores, at all stages of development, is pursued to improve image quality and sensitivity for functional imaging. So far our investigations with the casper mutants have enabled the identification of the specialized hearing organs, fluid-filled canal connecting the ears, and sub-structures of the semicircular canals. In our previous work with wild-type zebrafish, we were only able to identify and observe stimulated vibration of the largest structures, specifically the anterior swim bladder and tripus ossicle, even among small, larval specimen, with fully developed inner ears. In conclusion, this genetic mutant will enable the study of the dynamics of the zebrafish ear from the early larval stages all the way into adulthood.

  11. Facile Alkaline Lysis of Escherichia coli Cells in High-Throughput Mode for Screening Enzyme Mutants: Arylsulfatase as an Example.

    PubMed

    Yuan, Mei; Yang, Xiaolan; Li, Yuwei; Liu, Hongbo; Pu, Jun; Zhan, Chang-Guo; Liao, Fei

    2016-06-01

    Facile alkaline lysis of Escherichia coli cells in high-throughput (HTP) mode for screening enzyme mutants was tested with Pseudomonas aeruginosa arylsulfatase (PAAS). The alkaline lysis buffer was 1.0 M Tris-HCl at pH 9.0 plus 0.1 % Tween-20 and 2.0 mM 4-aminobenzamidine, mixed with cell suspension at 8:1 to 12:1 ratio for continuous agitation of mixtures in 96-well plates under room temperature; enzymatic activity in lysates was measured with 96-well microplate. PAAS activity tolerated final 0.1 % Tween-20. Individual clones were amplified for 12 h in 0.50 mL TB medium with 48-well plates to enhance the repeatability of induced expression. During continuous agitation of the mixture of cells and the lysis buffer, PAAS activities in lysates were steady from 3 to 9 h and comparable to sonication treatment but better than freezing-thawing. Coefficients of variation of activities of PAAS/mutants in lysates after treatment for 7 h reached ∼22 %. The mutant M72Q had specific activity 2-fold of G138S. By HTP lysis of cells, M72Q was recognized as a positive mutant over G138S with the area under the curve of 0.873. Therefore, for enzymes tolerating concentrated alkaline buffers, the proposed alkaline lysis approach may be generally applicable for HTP lysis of host cells during directed evolution. PMID:26899233

  12. Biochemical nature of cellulases from mutants of Trichoderma reesei

    SciTech Connect

    Montenecourt, B.S.; Kelleher, T.J.; Eveleigh, D.E.; Pettersson, L.G.

    1980-01-01

    The cellulases of two new mutants of T. reesei, RUT-NG14 and RUT-C30, have been examined with respect to the separation and biochemical characterization of the various components in the cellulase complex. The cellulase of both mutants has been shown to contain enhanced proportions of a cellobiohydrolase by quantitative immune precipitation. Application of ion-exchange high-pressure liquid chromatography protein separations affords a rapid (30 min) and simple method of separating and comparing cellulase components from potential high-yielding cellulase mutants.

  13. Attenuation of high sucrose diet–induced insulin resistance in tryptophan 2,3-dioxygenase deficient Drosophila melanogaster vermilion mutants

    PubMed Central

    Navrotskaya, Valeriya; Oxenkrug, Gregory; Vorobyova, Lyudmila; Summergrad, Paul

    2015-01-01

    Exposure to high sugar diet (HSD) serves as an experimental model of insulin resistance (IR) and type 2 diabetes (T2D) in mammals and insects. Peripheral IR induced by HSD delays emergence of pupae from larvae and decreases body weight of Drosophila imago. Understanding of mechanisms of IR/T2D is essential for refining T2D prevention and treatment strategies. Dysregulation of tryptophan (TRP) – kynurenine (KYN) pathway was suggested as one of the mechanisms of IR development. Rate-limiting enzyme of TRP – KYN pathway in Drosophila is TRP 2,3-dioxygenase (TDO), an evolutionary conserved ortholog of human TDO. In insects TDO is encoded by vermilion gene. TDO is not active in vermilion mutants. In order to evaluate the possible impact of deficient formation of KYN from TRP on the inducement of IR by HSD, we compared the effect of HSD in wild type (Oregon) and vermilion mutants of Drosophila melanogaster by assessing the time of white pupae emergence from larva and body weight of imago. Delay of emergence of pupae from larvae induced by high sucrose diet was less pronounced in vermilion (1.4 days) than in Oregon flies (3.3 days) in comparison with flies maintained on standard diet. Exposure to high sucrose diet decreased body weight of Oregon (but not vermilion) imago. Attenuation of high sucrose diet–induced IR/T2D in vermilion flies might depend on deficiency of TRP – KYN pathway. Besides IR/T2D, HSD induces obesity in Drosophila. Future studies of HSD-induced obesity and IR/T2D in TDO deficient vermilion mutants of Drosophila might help to understand the mechanisms of high association between IR/T2D and obesity. Modulation of TRP – KYN metabolism might be utilized for prevention and treatment of IR/T2D. PMID:26191458

  14. High fidelity simian immunodeficiency virus reverse transcriptase mutants have impaired replication in vitro and in vivo.

    PubMed

    Lloyd, Sarah B; Lichtfuss, Marit; Amarasena, Thakshila H; Alcantara, Sheilajen; De Rose, Robert; Tachedjian, Gilda; Alinejad-Rokny, Hamid; Venturi, Vanessa; Davenport, Miles P; Winnall, Wendy R; Kent, Stephen J

    2016-05-01

    The low fidelity of HIV replication facilitates immune and drug escape. Some reverse transcriptase (RT) inhibitor drug-resistance mutations increase RT fidelity in biochemical assays but their effect during viral replication is unclear. We investigated the effect of RT mutations K65R, Q151N and V148I on SIV replication and fidelity in vitro, along with SIV replication in pigtailed macaques. SIVmac239-K65R and SIVmac239-V148I viruses had reduced replication capacity compared to wild-type SIVmac239. Direct virus competition assays demonstrated a rank order of wild-type>K65R>V148I mutants in terms of viral fitness. In single round in vitro-replication assays, SIVmac239-K65R demonstrated significantly higher fidelity than wild-type, and rapidly reverted to wild-type following infection of macaques. In contrast, SIVmac239-Q151N was replication incompetent in vitro and in pigtailed macaques. Thus, we showed that RT mutants, and specifically the common K65R drug-resistance mutation, had impaired replication capacity and higher fidelity. These results have implications for the pathogenesis of drug-resistant HIV. PMID:26896929

  15. Genome-based establishment of a high-yielding heterotic pattern for hybrid wheat breeding

    PubMed Central

    Zhao, Yusheng; Li, Zuo; Liu, Guozheng; Jiang, Yong; Maurer, Hans Peter; Würschum, Tobias; Mock, Hans-Peter; Matros, Andrea; Ebmeyer, Erhard; Schachschneider, Ralf; Kazman, Ebrahim; Schacht, Johannes; Gowda, Manje; Longin, C. Friedrich H.; Reif, Jochen C.

    2015-01-01

    Hybrid breeding promises to boost yield and stability. The single most important element in implementing hybrid breeding is the recognition of a high-yielding heterotic pattern. We have developed a three-step strategy for identifying heterotic patterns for hybrid breeding comprising the following elements. First, the full hybrid performance matrix is compiled using genomic prediction. Second, a high-yielding heterotic pattern is searched based on a developed simulated annealing algorithm. Third, the long-term success of the identified heterotic pattern is assessed by estimating the usefulness, selection limit, and representativeness of the heterotic pattern with respect to a defined base population. This three-step approach was successfully implemented and evaluated using a phenotypic and genomic wheat dataset comprising 1,604 hybrids and their 135 parents. Integration of metabolomic-based prediction was not as powerful as genomic prediction. We show that hybrid wheat breeding based on the identified heterotic pattern can boost grain yield through the exploitation of heterosis and enhance recurrent selection gain. Our strategy represents a key step forward in hybrid breeding and is relevant for self-pollinating crops, which are currently shifting from pure-line to high-yielding and resilient hybrid varieties. PMID:26663911

  16. Genome-based establishment of a high-yielding heterotic pattern for hybrid wheat breeding.

    PubMed

    Zhao, Yusheng; Li, Zuo; Liu, Guozheng; Jiang, Yong; Maurer, Hans Peter; Würschum, Tobias; Mock, Hans-Peter; Matros, Andrea; Ebmeyer, Erhard; Schachschneider, Ralf; Kazman, Ebrahim; Schacht, Johannes; Gowda, Manje; Longin, C Friedrich H; Reif, Jochen C

    2015-12-22

    Hybrid breeding promises to boost yield and stability. The single most important element in implementing hybrid breeding is the recognition of a high-yielding heterotic pattern. We have developed a three-step strategy for identifying heterotic patterns for hybrid breeding comprising the following elements. First, the full hybrid performance matrix is compiled using genomic prediction. Second, a high-yielding heterotic pattern is searched based on a developed simulated annealing algorithm. Third, the long-term success of the identified heterotic pattern is assessed by estimating the usefulness, selection limit, and representativeness of the heterotic pattern with respect to a defined base population. This three-step approach was successfully implemented and evaluated using a phenotypic and genomic wheat dataset comprising 1,604 hybrids and their 135 parents. Integration of metabolomic-based prediction was not as powerful as genomic prediction. We show that hybrid wheat breeding based on the identified heterotic pattern can boost grain yield through the exploitation of heterosis and enhance recurrent selection gain. Our strategy represents a key step forward in hybrid breeding and is relevant for self-pollinating crops, which are currently shifting from pure-line to high-yielding and resilient hybrid varieties.

  17. On the possibility of long-wavelength long-lifetime high-quantum-yield luminophores.

    PubMed

    Lakowicz, J R; Piszczek, G; Kang, J S

    2001-01-01

    We describe an approach to creating a new class of luminophores which display both long wavelength emissions exceeding 600 nm and long lifetimes. These luminophores are based on resonance energy transfer (RET) from a long lifetime donor to a short lifetime but long wavelength acceptor. We demonstrated the possibility of obtaining these desirable spectral properties using donors and acceptors noncovalently bound to DNA. The donor was a ruthenium (Ru) metal-ligand complex in which one of the diimine ligands intercalated into double-helix DNA. The acceptors were either nile blue, TOTO-3, or TO-PRO-3. Upon binding of the acceptor to donor-labeled DNA, we found that the acceptor quantum yield was remarkably enhanced so that the wavelength-integrated intensities of the donor and acceptor bound to DNA were many-fold greater than the intensity of the donor and acceptor alone when separately bound to DNA. The origin of this effect is efficient energy transfer from the donor. Under these conditions the effective overall quantum yield approaches that of the acceptor. Importantly, the increased quantum yield can be obtained while maintaining usefully long apparent acceptor lifetimes of 30 to 80 ns. The effect of an increased quantum yield from a low quantum yield donor may find use in assays to detect macromolecular binding interactions. These results suggest the synthesis of covalently linked donor-acceptor pairs with the desirable spectral properties of long wavelength emission, high quantum yield, and moderately long lifetimes for gated detection.

  18. Mass production of multi-wall carbon nanotubes by metal dusting process with high yield

    SciTech Connect

    Ghorbani, H.; Rashidi, A.M.; Rastegari, S.; Mirdamadi, S.; Alaei, M.

    2011-05-15

    Research highlights: {yields} Synthesis of carbon nanotubes over Fe-Ni nanoparticles supported alloy 304L. {yields} Production of carbon nanotubes with high yield (700-1000%) and low cost catalyst. {yields} Optimum growth condition is CO/H{sub 2} = 1/1, 100 cm{sup 3}/min, at 620 {sup o}C under long term repetitive thermal cycling. {yields} Possibility of the mass production by metal dusting process with low cost. -- Abstract: Carbon nanotube materials were synthesized over Fe-Ni nanoparticles generated during disintegration of the surface of alloy 304L under metal dusting environment. The metal dusting condition was simulated and optimized through exposing stainless steel samples during long term repetitive thermal cycling in CO/H{sub 2} = 1/1, total gas flow rate 100 cm{sup 3}/min, at 620 {sup o}C for 300 h. After reaction, surface morphology of the samples and also carbonaceous deposition which had grown on sample surfaces were examined by stereoscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Results revealed that multi-wall carbon nanotubes could be formed over nanocatalyst generated on the alloy surface by exploiting metal dusting process. By optimization of reaction parameters the yields of carbon nanotube materials obtained were 700-1000%. Also it has been shown herein that the amount of carbon nanotube materials remarkably increases when the reaction time is extended up to 300 h, indicating a possibility of the mass production by this easy method.

  19. Kinetic characterization of high-activity mutants of human butyrylcholinesterase for the cocaine metabolite norcocaine.

    PubMed

    Zhan, Max; Hou, Shurong; Zhan, Chang-Guo; Zheng, Fang

    2014-01-01

    It has been known that cocaine produces its toxic and physiological effects through not only cocaine itself, but also norcocaine formed from cocaine oxidation catalysed by microsomal CYP (cytochrome P450) 3A4 in the human liver. The catalytic parameters (kcat and Km) of human BChE (butyrylcholinesterase) and its three mutants (i.e. A199S/S287G/A328W/Y332G, A199S/F227A/S287G/A328W/E441D and A199S/F227A/S287G/A328W/Y332G) for norcocaine have been characterized in the present study for the first time and compared with those for cocaine. On the basis of the obtained kinetic data, wild-type human BChE has a significantly lower catalytic activity for norcocaine (kcat=2.8 min(-1), Km=15 μM and kcat/Km=1.87 × 10(5) M(-1)·min(-1)) compared with its catalytic activity for (-)-cocaine. The BChE mutants examined in the present study have considerably improved catalytic activities against both cocaine and norcocaine compared with the wild-type enzyme. Within the enzymes examined in the present study, the A199S/F227A/S287G/A328W/Y332G mutant (CocH3) is identified as the most efficient enzyme for hydrolysing both cocaine and norcocaine. CocH3 has a 1080-fold improved catalytic efficiency for norcocaine (kcat=2610 min(-1), Km=13 μM and kcat/Km=2.01 × 10(8) M(-1)·min(-1)) and a 2020-fold improved catalytic efficiency for cocaine. It has been demonstrated that CocH3 as an exogenous enzyme can rapidly metabolize norcocaine, in addition to cocaine, in rats. Further kinetic modelling has suggested that CocH3 with an identical concentration with that of the endogenous BChE in human plasma can effectively eliminate both cocaine and norcocaine in a simplified kinetic model of cocaine abuse.

  20. Molecular determinants for the high constitutive activity of the human histamine H4 receptor: functional studies on orthologues and mutants

    PubMed Central

    Wifling, D; Löffel, K; Nordemann, U; Strasser, A; Bernhardt, G; Dove, S; Seifert, R; Buschauer, A

    2015-01-01

    Background and Purpose Some histamine H4 receptor ligands act as inverse agonists at the human H4 receptor (hH4R), a receptor with exceptionally high constitutive activity, but as neutral antagonists or partial agonists at the constitutively inactive mouse H4 receptor (mH4R) and rat H4 receptor (rH4R). To study molecular determinants of constitutive activity, H4 receptor reciprocal mutants were constructed: single mutants: hH4R-F169V, mH4R-V171F, hH4R-S179A, hH4R-S179M; double mutants: hH4R-F169V+S179A, hH4R-F169V+S179M and mH4R-V171F+M181S. Experimental Approach Site-directed mutagenesis with pVL1392 plasmids containing hH4 or mH4 receptors were performed. Wild-type or mutant receptors were co-expressed with Gαi2 and Gβ1γ2 in Sf9 cells. Membranes were studied in saturation and competition binding assays ([3H]-histamine), and in functional [35S]-GTPγS assays with inverse, partial and full agonists of the hH4 receptor. Key Results Constitutive activity decreased from the hH4 receptor via the hH4R-F169V mutant to the hH4R-F169V+S179A and hH4R-F169V+S179M double mutants. F169 alone or in concert with S179 plays a major role in stabilizing a ligand-free active state of the hH4 receptor. Partial inverse hH4 receptor agonists like JNJ7777120 behaved as neutral antagonists or partial agonists at species orthologues with lower or no constitutive activity. Some partial and full hH4 receptor agonists showed decreased maximal effects and potencies at hH4R-F169V and double mutants. However, the mutation of S179 in the hH4 receptor to M as in mH4 receptor or A as in rH4 receptor did not significantly reduce constitutive activity. Conclusions and Implications F169 and S179 are key amino acids for the high constitutive activity of hH4 receptors and may also be of relevance for other constitutively active GPCRs. Linked Articles This article is part of a themed issue on Histamine Pharmacology Update published in volume 170 issue 1. To view the other articles in this issue visit

  1. The diageotropica mutant of tomato lacks high specific activity auxin sites

    SciTech Connect

    Hicks, G.R.; Lomax, T.L. ); Rayle, D.L. )

    1989-04-01

    Tomato (Lycopersicum esculentum, Mill) plants homozygous for the single gene diageotropica (dgt) mutation have reduced shoot growth, abnormal vascular tissue, altered leaf morphology, and lack of lateral root branching. These and other morphological and physiological abnormalities suggest that dgt plants are unable to respond to the plant growth hormone auxin (indole-3-acetic acid, IAA). The photoaffinity auxin analogue {sup 3}H-5N{sub 3}-IAA specifically labels a polypeptide doublet of 40 ad 42 kD in membrane preparations from stems of the parental variety VFN8, but not from stems of dgt. In elongation tests, excised dgt roots respond in the same manner to IAA an VFN8 roots. These data suggest that the two polypeptides are part of a physiologically important auxin receptor system which is altered in a tissue-specific manner in the mutant.

  2. High-resolution, regional-scale crop yield simulations for the Southwestern United States

    NASA Astrophysics Data System (ADS)

    Stack, D. H.; Kafatos, M.; Medvigy, D.; El-Askary, H. M.; Hatzopoulos, N.; Kim, J.; Kim, S.; Prasad, A. K.; Tremback, C.; Walko, R. L.; Asrar, G. R.

    2012-12-01

    Over the past few decades, there have been many process-based crop models developed with the goal of better understanding the impacts of climate, soils, and management decisions on crop yields. These models simulate the growth and development of crops in response to environmental drivers. Traditionally, process-based crop models have been run at the individual farm level for yield optimization and management scenario testing. Few previous studies have used these models over broader geographic regions, largely due to the lack of gridded high-resolution meteorological and soil datasets required as inputs for these data intensive process-based models. In particular, assessment of regional-scale yield variability due to climate change requires high-resolution, regional-scale, climate projections, and such projections have been unavailable until recently. The goal of this study was to create a framework for extending the Agricultural Production Systems sIMulator (APSIM) crop model for use at regional scales and analyze spatial and temporal yield changes in the Southwestern United States (CA, AZ, and NV). Using the scripting language Python, an automated pipeline was developed to link Regional Climate Model (RCM) output with the APSIM crop model, thus creating a one-way nested modeling framework. This framework was used to combine climate, soil, land use, and agricultural management datasets in order to better understand the relationship between climate variability and crop yield at the regional-scale. Three different RCMs were used to drive APSIM: OLAM, RAMS, and WRF. Preliminary results suggest that, depending on the model inputs, there is some variability between simulated RCM driven maize yields and historical yields obtained from the United States Department of Agriculture (USDA). Furthermore, these simulations showed strong non-linear correlations between yield and meteorological drivers, with critical threshold values for some of the inputs (e.g. minimum and

  3. Measurements of relative binding of cohesin and dockerin mutants using an advanced ELISA technique for high-affinity interactions.

    PubMed

    Slutzki, Michal; Barak, Yoav; Reshef, Dan; Schueler-Furman, Ora; Lamed, Raphael; Bayer, Edward A

    2012-01-01

    The cellulosome is a large bacterial extracellular multienzyme complex able to degrade crystalline cellulosic substrates. The complex contains catalytic and noncatalytic subunits, interconnected by high-affinity cohesin-dockerin interactions. In this chapter, we introduce an optimized method for comparative binding among different cohesins or cohesin mutants to the dockerin partner. This assay offers advantages over other methods (such as ELISA, cELIA, SPR, and ITC) for particularly high-affinity binding interactions. In this approach, the high-affinity interaction of interest occurs in the liquid phase during the equilibrated binding step, whereas the interaction with the immobilized phase is used only for detection of the unbound dockerins that remain in the solution phase. Once equilibrium conditions are reached, the change in free energy of binding (ΔΔG(binding)), as well as the affinity constant of mutants, can be estimated against the known affinity constant of the wild-type interaction. In light of the above, we propose this method as a preferred alternative for the relative quantification of high-affinity protein interactions. PMID:22608739

  4. Yield-determining factors in high-solids enzymatic hydrolysis of lignocellulose

    PubMed Central

    Kristensen, Jan B; Felby, Claus; Jørgensen, Henning

    2009-01-01

    Background Working at high solids (substrate) concentrations is advantageous in enzymatic conversion of lignocellulosic biomass as it increases product concentrations and plant productivity while lowering energy and water input. However, for a number of lignocellulosic substrates it has been shown that at increasing substrate concentration, the corresponding yield decreases in a fashion which can not be explained by current models and knowledge of enzyme-substrate interactions. This decrease in yield is undesirable as it offsets the advantages of working at high solids levels. The cause of the 'solids effect' has so far remained unknown. Results The decreasing conversion at increasing solids concentrations was found to be a generic or intrinsic effect, describing a linear correlation from 5 to 30% initial total solids content (w/w). Insufficient mixing has previously been shown not to be involved in the effect. Hydrolysis experiments with filter paper showed that neither lignin content nor hemicellulose-derived inhibitors appear to be responsible for the decrease in yields. Product inhibition by glucose and in particular cellobiose (and ethanol in simultaneous saccharification and fermentation) at the increased concentrations at high solids loading plays a role but could not completely account for the decreasing conversion. Adsorption of cellulases was found to decrease at increasing solids concentrations. There was a strong correlation between the decreasing adsorption and conversion, indicating that the inhibition of cellulase adsorption to cellulose is causing the decrease in yield. Conclusion Inhibition of enzyme adsorption by hydrolysis products appear to be the main cause of the decreasing yields at increasing substrate concentrations in the enzymatic decomposition of cellulosic biomass. In order to facilitate high conversions at high solids concentrations, understanding of the mechanisms involved in high-solids product inhibition and adsorption inhibition

  5. NEW ACALA GERMPLASM LINES WITH HIGH YIELD POTENTIAL AND FIBER QUALITY

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This paper reports the development of 18 Acala 1517 cotton germplasm lines developed in the New Mexico Cotton Breeding Program. NM W1218 is an Acala cotton line with okra leaf and 7.8% higher lint yield than Acala 1517-99. NM 970123 is an Acala cotton line with high lint percentage (44.6%) and 6.4...

  6. Comparing high density LIDAR and medium resolution GPS generated elevation data for predicting yield stability

    Technology Transfer Automated Retrieval System (TEKTRAN)

    High density light detection and ranging (LIDAR) imaging has been shown to be able to define yield stability areas of a field for multi-cropping. Since LIDAR imaging is expensive and not widely available, we hypothesized that medium resolution GPS elevation data which is commonly collected with var...

  7. High yield synthesis of high-silica chabazite by combining the role of zeolite precursors and tetraethylammonium: SCR of NOx.

    PubMed

    Martín, Nuria; Moliner, Manuel; Corma, Avelino

    2015-06-21

    The synthesis of chabazite with high solid yields is achieved by the rational combination of directing effects of a source of Si and Al coming from USY zeolites and the inexpensive tetraethylammonium. Moreover, Cu-CHA materials prepared by post-synthetic and "one-pot" methodologies show high activity and stability for SCR of NOx.

  8. Canopy Light Interception of a Conventional and an Erect Leaf Mutant Sorghum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two sorghum lines, an erect leafed mutant sorghum and the wild type from which the mutant was generated, were field grown in rectilinear arrays at low (23 plants per square meter) and high (10 plants per square meter) population densities. Canopy light interception, biomass accretion and yield were ...

  9. Repair effects of laser on mutants of filamentous fungi

    NASA Astrophysics Data System (ADS)

    Zhao, Yansheng; Xiao, Canpeng; Qian, Hailun; Su, Baoliang; Hu, Yujun; Deng, Jianhui

    1999-09-01

    The paper reports that penicillin-producing strains and lovastatin-producing strains were irradiated by UV and subsequently by laser (632.8 nm), and the reparation rate reached 297% and 264%. High-yield mutant was selected with improved potency of 24.5% and 30%, respectively; Gibberellin producing strains were treated with chemical agent LiCl, and then irradiated with 632.8 nm laser. One mutant with 189.6% increased potency was obtained. The experimental results indicated that using laser irradiation after UV or chemical agent mutation was a new useful method in breeding high-yield strains.

  10. High-yield reactivation of anionic tobacco peroxidase overexpressed in Escherichia coli.

    PubMed

    Zakharova, G S; Poloznikov, A A; Chubar, T A; Gazaryan, I G; Tishkov, V I

    2015-09-01

    Anionic tobacco peroxidase (TOP) is extremely active in chemiluminescence reaction of luminol oxidation without addition of enhancers and more stable than horseradish peroxidase under antibody conjugation conditions. In addition, recombinant TOP (rTOP) produced in Escherichia coli is known to be a perfect direct electron transfer catalyst on electrodes of various origin. These features make the task of development of a high-yield reactivation protocol for rTOP practically important. Previous attempts to reactivate the enzyme from E. coli inclusion bodies were successful, but the reported reactivation yield was only 14%. In this work, we thoroughly screened the refolding conditions for dilution protocol and compared it with gel-filtration chromatography. The impressive reactivation yield in the dilution protocol (85%) was achieved for 8 μg/mL solubilized rTOP protein and the refolding medium containing 0.3 mM oxidized glutathione, 0.05 mM dithiothreitol, 5 mM CaCl2, 5% glycerol in 50 mM Tris-HCl buffer, pH 9.6, with 1 μM hemin added at the 24th hour of incubation. A practically important discovery was a 30-40% increase in the reactivation yield upon delayed addition of hemin. The reactivation yield achieved is one of the highest reported in the literature on protein refolding by dilution. The final yield of purified active non-glycosylated rTOP was ca. 60 mg per L of E. coli culture, close to the yield reported before for tomato and tobacco plants overexpressing glycosylated TOP (60 mg/kg biomass) and much higher than for the previously reported refolding protocol (2.6 mg per L of E. coli culture).

  11. Microbial electrolysis cells for high yield hydrogen gas production from organic matter.

    PubMed

    Logan, Bruce E; Call, Douglas; Cheng, Shaoan; Hamelers, Hubertus V M; Sleutels, Tom H J A; Jeremiasse, Adriaan W; Rozendal, René A

    2008-12-01

    The use of electrochemically active bacteria to break down organic matter, combined with the addition of a small voltage (> 0.2 V in practice) in specially designed microbial electrolysis cells (MECs), can result in a high yield of hydrogen gas. While microbial electrolysis was invented only a few years ago, rapid developments have led to hydrogen yields approaching 100%, energy yields based on electrical energy input many times greater than that possible by water electrolysis, and increased gas production rates. MECs used to make hydrogen gas are similar in design to microbial fuel cells (MFCs) that produce electricity, but there are important differences in architecture and analytical methods used to evaluate performance. We review here the materials, architectures, performance, and energy efficiencies of these MEC systems that show promise as a method for renewable and sustainable energy production, and wastewater treatment.

  12. Differential metabolite profiles during fruit development in high-yielding oil palm mesocarp.

    PubMed

    Teh, Huey Fang; Neoh, Bee Keat; Hong, May Ping Li; Low, Jaime Yoke Sum; Ng, Theresa Lee Mei; Ithnin, Nalisha; Thang, Yin Mee; Mohamed, Mohaimi; Chew, Fook Tim; Yusof, Hirzun Mohd; Kulaveerasingam, Harikrishna; Appleton, David R

    2013-01-01

    To better understand lipid biosynthesis in oil palm mesocarp, in particular the differences in gene regulation leading to and including de novo fatty acid biosynthesis, a multi-platform metabolomics technology was used to profile mesocarp metabolites during six critical stages of fruit development in comparatively high- and low-yielding oil palm populations. Significantly higher amino acid levels preceding lipid biosynthesis and nucleosides during lipid biosynthesis were observed in a higher yielding commercial palm population. Levels of metabolites involved in glycolysis revealed interesting divergence of flux towards glycerol-3-phosphate, while carbon utilization differences in the TCA cycle were proven by an increase in malic acid/citric acid ratio. Apart from insights into the regulation of enhanced lipid production in oil palm, these results provide potentially useful metabolite yield markers and genes of interest for use in breeding programmes. PMID:23593468

  13. High Photoluminescence Quantum Yield in Band Gap Tunable Bromide Containing Mixed Halide Perovskites.

    PubMed

    Sutter-Fella, Carolin M; Li, Yanbo; Amani, Matin; Ager, Joel W; Toma, Francesca M; Yablonovitch, Eli; Sharp, Ian D; Javey, Ali

    2016-01-13

    Hybrid organic-inorganic halide perovskite based semiconductor materials are attractive for use in a wide range of optoelectronic devices because they combine the advantages of suitable optoelectronic attributes and simultaneously low-cost solution processability. Here, we present a two-step low-pressure vapor-assisted solution process to grow high quality homogeneous CH3NH3PbI3-xBrx perovskite films over the full band gap range of 1.6-2.3 eV. Photoluminescence light-in versus light-out characterization techniques are used to provide new insights into the optoelectronic properties of Br-containing hybrid organic-inorganic perovskites as a function of optical carrier injection by employing pump-powers over a 6 orders of magnitude dynamic range. The internal luminescence quantum yield of wide band gap perovskites reaches impressive values up to 30%. This high quantum yield translates into substantial quasi-Fermi level splitting and high "luminescence or optically implied" open-circuit voltage. Most importantly, both attributes, high internal quantum yield and high optically implied open-circuit voltage, are demonstrated over the entire band gap range (1.6 eV ≤ Eg ≤ 2.3 eV). These results establish the versatility of Br-containing perovskite semiconductors for a variety of applications and especially for the use as high-quality top cell in tandem photovoltaic devices in combination with industry dominant Si bottom cells. PMID:26691065

  14. High-material yield fabrication of YBCO coated conductors by Nd:YAG-PLD system

    NASA Astrophysics Data System (ADS)

    Ono, I.; Ichino, Y.; Yoshida, Y.; Yoshizumi, M.; Izumi, T.; Shiohara, Y.

    In order to fabricate superconducting coated conductors with lower cost, fabrication processes are required high material yield. We report an improvement of the material yield of YBCO films prepared by inside-plume Nd:YAG pulsed laser deposition method on metal substrates with an architecture of CeO2/LMO/IBAD-MgO/GZO/HastelloyTM. In this study, we shortened a distance from the target to the substrate (dT-S) in order to improve the material yield. Additionally, we have used Nd:YAG laser because initial and running costs are anticipated to be lower than those of excimer laser. As a result, by shorting the dT-S from 40 mm to 20 mm, the material yield increased on 10 mm×10 mm substrates. Additionally, by changing the O2 pressure (PO2) from 40 Pa to 400 Pa at dT-S = 20 mm, the material yield had a local maximal value of 18.6% at PO2=200 Pa. On multi-turn (MT) metal substrates, the material yield reached 56.0%. However, the critical current density (Jc) of the YBCO film which was deposited at dT-S = 20 mm and PO2=200 Pa on the 10 mm×10 mm substrate was 1.2 MA/cm2 at the edges and 0.2 MA/cm2 at the center at 77 K in self-field, although the YBCO films showed good aligned crystal textures. In order to improve this non-uniformity, we tilted the surface normal of the target 15 degree from a line connecting target and substrate. As a result, we achieved the uniformity in deposition rate. Then, we prepared YBa1.78Cu2.9Oy target to fabricate YBCO films with stoichiometric composition and obtained the uniform-high Jc.

  15. A baker's yeast mutant (fil1) with a specific, partially inactivating mutation in adenylate cyclase maintains a high stress resistance during active fermentation and growth.

    PubMed

    Van Dijck, P; Ma, P; Versele, M; Gorwa, M F; Colombo, S; Lemaire, K; Bossi, D; Loïez, A; Thevelein, J M

    2000-10-01

    The initiation of fermentation in the yeast Saccharomyces cerevisiae is associated with a rapid drop in stress resistance. This is disadvantageous for several biotechnological applications, e.g. the preparation of freeze doughs. We have isolated mutants in a laboratory strain which are deficient in fermentation-induced loss of stress resistance ('fil' mutants) using a heat shock selection protocol. We show that the fil1 mutant contains a mutation in the CYR1 gene which encodes adenylate cyclase. It causes a change at position 1682 of glutamate into lysine and results in a tenfold drop in adenylate cyclase activity. The fil1 mutant displays a reduction in the glucose-induced cAMP increase, trehalase activation and loss of heat resistance. Interestingly, the fil1 mutant shows the same growth and fermentation rate as the wild type strain, as opposed to other mutants with reduced activity of the cAMP pathway. Introduction of the fil1 mutation in the vigorous Y55 strain and cultivation of the mutant under pilot scale conditions resulted in a yeast that displayed a higher freeze and drought resistance during active fermentation compared to the wild type Y55 strain. These results show that high stress resistance and high fermentation activity are compatible biological properties. Isolation of fil-type mutations appears a promising avenue for development of industrial yeast strains with improved stress resistance during active fermentation.

  16. A baker's yeast mutant (fil1) with a specific, partially inactivating mutation in adenylate cyclase maintains a high stress resistance during active fermentation and growth.

    PubMed

    Van Dijck, P; Ma, P; Versele, M; Gorwa, M F; Colombo, S; Lemaire, K; Bossi, D; Loïez, A; Thevelein, J M

    2000-10-01

    The initiation of fermentation in the yeast Saccharomyces cerevisiae is associated with a rapid drop in stress resistance. This is disadvantageous for several biotechnological applications, e.g. the preparation of freeze doughs. We have isolated mutants in a laboratory strain which are deficient in fermentation-induced loss of stress resistance ('fil' mutants) using a heat shock selection protocol. We show that the fil1 mutant contains a mutation in the CYR1 gene which encodes adenylate cyclase. It causes a change at position 1682 of glutamate into lysine and results in a tenfold drop in adenylate cyclase activity. The fil1 mutant displays a reduction in the glucose-induced cAMP increase, trehalase activation and loss of heat resistance. Interestingly, the fil1 mutant shows the same growth and fermentation rate as the wild type strain, as opposed to other mutants with reduced activity of the cAMP pathway. Introduction of the fil1 mutation in the vigorous Y55 strain and cultivation of the mutant under pilot scale conditions resulted in a yeast that displayed a higher freeze and drought resistance during active fermentation compared to the wild type Y55 strain. These results show that high stress resistance and high fermentation activity are compatible biological properties. Isolation of fil-type mutations appears a promising avenue for development of industrial yeast strains with improved stress resistance during active fermentation. PMID:11075928

  17. Superior triacylglycerol (TAG) accumulation in starchless mutants of Scenedesmus obliquus: (I) mutant generation and characterization

    PubMed Central

    2014-01-01

    Background Microalgae are a promising platform for producing neutral lipids, to be used in the application for biofuels or commodities in the feed and food industry. A very promising candidate is the oleaginous green microalga Scenedesmus obliquus, because it accumulates up to 45% w/w triacylglycerol (TAG) under nitrogen starvation. Under these conditions, starch is accumulated as well. Starch can amount up to 38% w/w under nitrogen starvation, which is a substantial part of the total carbon captured. When aiming for optimized TAG production, blocking the formation of starch could potentially increase carbon allocation towards TAG. In an attempt to increase TAG content, productivity and yield, starchless mutants of this high potential strain were generated using UV mutagenesis. Previous studies in Chlamydomonas reinhardtii have shown that blocking the starch synthesis yields higher TAG contents, although these TAG contents do not surpass those of oleaginous microalgae yet. So far no starchless mutants in oleaginous green microalgae have been isolated that result in higher TAG productivities. Results Five starchless mutants have been isolated successfully from over 3,500 mutants. The effect of the mutation on biomass and total fatty acid (TFA) and TAG productivity under nitrogen-replete and nitrogen-depleted conditions was studied. All five starchless mutants showed a decreased or completely absent starch content. In parallel, an increased TAG accumulation rate was observed for the starchless mutants and no substantial decrease in biomass productivity was perceived. The most promising mutant showed an increase in TFA productivity of 41% at 4 days after nitrogen depletion, reached a TAG content of 49.4% (% of dry weight) and had no substantial change in biomass productivity compared to the wild type. Conclusions The improved S. obliquus TAG production strains are the first starchless mutants in an oleaginous green microalga that show enhanced TAG content under

  18. A defective signal peptide in the maize high-lysine mutant floury 2.

    PubMed Central

    Coleman, C E; Lopes, M A; Gillikin, J W; Boston, R S; Larkins, B A

    1995-01-01

    The maize floury 2 (fl2) mutation enhances the lysine content of the grain, but the soft texture of the endosperm makes it unsuitable for commercial production. The mutant phenotype is linked with the appearance of a 24-kDa alpha-zein protein and increased synthesis of binding protein, both of which are associated with irregularly shaped protein bodies. We have cloned the gene encoding the 24-kDa protein and show that it is expressed as a 22-kDa alpha-zein with an uncleaved signal peptide. Comparison of the deduced N-terminal amino acid sequence of the 24-kDa alpha-zein protein with other alpha-zeins revealed an alanine to valine substitution at the C-terminal position of the signal peptide, a histidine insertion within the seventh alpha-helical repeat, and an alanine to threonine substitution with the same alpha-helical repeat of the protein. Structural defects associated with this alpha-zein explain many of the phenotypic effects of the fl2 mutation. Images Fig. 1 Fig. 2 Fig. 5 PMID:7624327

  19. Assessing mutant p53 in primary high-grade serous ovarian cancer using immunohistochemistry and massively parallel sequencing

    PubMed Central

    Cole, Alexander J.; Dwight, Trisha; Gill, Anthony J.; Dickson, Kristie-Ann; Zhu, Ying; Clarkson, Adele; Gard, Gregory B.; Maidens, Jayne; Valmadre, Susan; Clifton-Bligh, Roderick; Marsh, Deborah J.

    2016-01-01

    The tumour suppressor p53 is mutated in cancer, including over 96% of high-grade serous ovarian cancer (HGSOC). Mutations cause loss of wild-type p53 function due to either gain of abnormal function of mutant p53 (mutp53), or absent to low mutp53. Massively parallel sequencing (MPS) enables increased accuracy of detection of somatic variants in heterogeneous tumours. We used MPS and immunohistochemistry (IHC) to characterise HGSOCs for TP53 mutation and p53 expression. TP53 mutation was identified in 94% (68/72) of HGSOCs, 62% of which were missense. Missense mutations demonstrated high p53 by IHC, as did 35% (9/26) of non-missense mutations. Low p53 was seen by IHC in 62% of HGSOC associated with non-missense mutations. Most wild-type TP53 tumours (75%, 6/8) displayed intermediate p53 levels. The overall sensitivity of detecting a TP53 mutation based on classification as ‘Low’, ‘Intermediate’ or ‘High’ for p53 IHC was 99%, with a specificity of 75%. We suggest p53 IHC can be used as a surrogate marker of TP53 mutation in HGSOC; however, this will result in misclassification of a proportion of TP53 wild-type and mutant tumours. Therapeutic targeting of mutp53 will require knowledge of both TP53 mutations and mutp53 expression. PMID:27189670

  20. Flow “Fine” Synthesis: High Yielding and Selective Organic Synthesis by Flow Methods

    PubMed Central

    2015-01-01

    Abstract The concept of flow “fine” synthesis, that is, high yielding and selective organic synthesis by flow methods, is described. Some examples of flow “fine” synthesis of natural products and APIs are discussed. Flow methods have several advantages over batch methods in terms of environmental compatibility, efficiency, and safety. However, synthesis by flow methods is more difficult than synthesis by batch methods. Indeed, it has been considered that synthesis by flow methods can be applicable for the production of simple gasses but that it is difficult to apply to the synthesis of complex molecules such as natural products and APIs. Therefore, organic synthesis of such complex molecules has been conducted by batch methods. On the other hand, syntheses and reactions that attain high yields and high selectivities by flow methods are increasingly reported. Flow methods are leading candidates for the next generation of manufacturing methods that can mitigate environmental concerns toward sustainable society. PMID:26337828

  1. Flow "Fine" Synthesis: High Yielding and Selective Organic Synthesis by Flow Methods.

    PubMed

    Kobayashi, Shū

    2016-02-18

    The concept of flow "fine" synthesis, that is, high yielding and selective organic synthesis by flow methods, is described. Some examples of flow "fine" synthesis of natural products and APIs are discussed. Flow methods have several advantages over batch methods in terms of environmental compatibility, efficiency, and safety. However, synthesis by flow methods is more difficult than synthesis by batch methods. Indeed, it has been considered that synthesis by flow methods can be applicable for the production of simple gasses but that it is difficult to apply to the synthesis of complex molecules such as natural products and APIs. Therefore, organic synthesis of such complex molecules has been conducted by batch methods. On the other hand, syntheses and reactions that attain high yields and high selectivities by flow methods are increasingly reported. Flow methods are leading candidates for the next generation of manufacturing methods that can mitigate environmental concerns toward sustainable society.

  2. Comparative transcriptome analysis using high papaverine mutant of Papaver somniferum reveals pathway and uncharacterized steps of papaverine biosynthesis.

    PubMed

    Pathak, Sumya; Lakhwani, Deepika; Gupta, Parul; Mishra, Brij Kishore; Shukla, Sudhir; Asif, Mehar Hasan; Trivedi, Prabodh Kumar

    2013-01-01

    The benzylisoquinoline alkaloid papaverine, synthesized in low amount in most of the opium poppy varieties of Papaver somniferum, is used as a vasodilator muscle relaxant and antispasmodic. Papaverine biosynthesis remains controversial as two different routes utilizing either (S)-coclaurine or (S)-reticuline have been proposed with uncharacterized intermediate steps. In an attempt to elucidate papaverine biosynthesis and identify putative genes involved in uncharacterized steps, we carried out comparative transcriptome analysis of high papaverine mutant (pap1) and normal cultivar (BR086) of P. somniferum. This natural mutant synthesizes more than 12-fold papaverine in comparison to BR086. We established more than 238 Mb transcriptome data separately for pap1 and BR086. Assembly of reads generated 127,342 and 106,128 unigenes in pap1 and BR086, respectively. Digital gene expression analysis of transcriptomes revealed 3,336 differentially expressing unigenes. Enhanced expression of (S)-norcoclaurine-6-O-methyltransferase (6OMT), (S)-3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase (4'OMT), norreticuline 7-O-methyltransferase (N7OMT) and down-regulation of reticuline 7-O-methyltransferase (7OMT) in pap1 in comparison to BR086 suggest (S)-coclaurine as the route for papaverine biosynthesis. We also identified several methyltransferases and dehydrogenases with enhanced expression in pap1 in comparison to BR086. Our analysis using natural mutant, pap1, concludes that (S)-coclaurine is the branch-point intermediate and preferred route for papaverine biosynthesis. Differentially expressing methyltransferases and dehydrogenases identified in this study will help in elucidating complete biosynthetic pathway of papaverine. The information generated will be helpful in developing strategies for enhanced biosynthesis of papaverine through biotechnological approaches.

  3. A novel high-throughput in vivo molecular screen for shade avoidance mutants identifies a novel phyA mutation.

    PubMed

    Wang, Xuewen; Roig-Villanova, Irma; Khan, Safina; Shanahan, Hugh; Quail, Peter H; Martinez-Garcia, Jaime F; Devlin, Paul F

    2011-05-01

    The shade avoidance syndrome (SAS) allows plants to anticipate and avoid shading by neighbouring plants by initiating an elongation growth response. The phytochrome photoreceptors are able to detect a reduction in the red:far red ratio in incident light, the result of selective absorption of red and blue wavelengths by proximal vegetation. A shade-responsive luciferase reporter line (PHYB::LUC) was used to carry out a high-throughput screen to identify novel SAS mutants. The dracula 1 (dra1) mutant, that showed no avoidance of shade for the PHYB::LUC response, was the result of a mutation in the PHYA gene. Like previously characterized phyA mutants, dra1 showed a long hypocotyl in far red light and an enhanced hypocotyl elongation response to shade. However, dra1 additionally showed a long hypocotyl in red light. Since phyB levels are relatively unaffected in dra1, this gain-of-function red light phenotype strongly suggests a disruption of phyB signalling. The dra1 mutation, G773E within the phyA PAS2 domain, occurs at a residue absolutely conserved among phyA sequences. The equivalent residue in phyB is absolutely conserved as a threonine. PAS domains are structurally conserved domains involved in molecular interaction. Structural modelling of the dra1 mutation within the phyA PAS2 domain shows some similarity with the structure of the phyB PAS2 domain, suggesting that the interference with phyB signalling may be the result of non-functional mimicry. Hence, it was hypothesized that this PAS2 residue forms a key distinction between the phyA and phyB phytochrome species.

  4. A novel high-throughput in vivo molecular screen for shade avoidance mutants identifies a novel phyA mutation

    PubMed Central

    Roig-Villanova, Irma; Khan, Safina; Shanahan, Hugh; Quail, Peter H.; Martinez-Garcia, Jaime F.; Devlin, Paul F.

    2011-01-01

    The shade avoidance syndrome (SAS) allows plants to anticipate and avoid shading by neighbouring plants by initiating an elongation growth response. The phytochrome photoreceptors are able to detect a reduction in the red:far red ratio in incident light, the result of selective absorption of red and blue wavelengths by proximal vegetation. A shade-responsive luciferase reporter line (PHYB::LUC) was used to carry out a high-throughput screen to identify novel SAS mutants. The dracula 1 (dra1) mutant, that showed no avoidance of shade for the PHYB::LUC response, was the result of a mutation in the PHYA gene. Like previously characterized phyA mutants, dra1 showed a long hypocotyl in far red light and an enhanced hypocotyl elongation response to shade. However, dra1 additionally showed a long hypocotyl in red light. Since phyB levels are relatively unaffected in dra1, this gain-of-function red light phenotype strongly suggests a disruption of phyB signalling. The dra1 mutation, G773E within the phyA PAS2 domain, occurs at a residue absolutely conserved among phyA sequences. The equivalent residue in phyB is absolutely conserved as a threonine. PAS domains are structurally conserved domains involved in molecular interaction. Structural modelling of the dra1 mutation within the phyA PAS2 domain shows some similarity with the structure of the phyB PAS2 domain, suggesting that the interference with phyB signalling may be the result of non-functional mimicry. Hence, it was hypothesized that this PAS2 residue forms a key distinction between the phyA and phyB phytochrome species. PMID:21398429

  5. Ultra-broadband nonlinear saturable absorption of high-yield MoS2 nanosheets.

    PubMed

    Wei, Rongfei; Zhang, Hang; Hu, Zhongliang; Qiao, Tian; He, Xin; Guo, Qiangbing; Tian, Xiangling; Chen, Zhi; Qiu, Jianrong

    2016-07-29

    High-yield MoS2 nanosheets with strong nonlinear optical (NLO) responses in a broad near-infrared range were synthesized by a facile hydrothermal method. The observation of saturable absorption, which was excited by the light with photon energy smaller than the gap energy of MoS2, can be attributed to the enhancement of the hybridization between the Mo d-orbital and S p-orbital by the oxygen incorporation into MoS2. High-yield MoS2 nanosheets with high modulation depth and large saturable intensity generated a stable, passively Q-switched fiber laser pulse at 1.56 μm. The high output power of 1.08 mW can be attained under a very low pump power of 30.87 mW. Compared to recently reported passively Q-switched fiber lasers utilizing exfoliated MoS2 nanosheets, the efficiency of the laser for our passive Q-switching operation is larger and reaches 3.50%. This research may extend the understanding on the NLO properties of MoS2 and indicate the feasibility of the high-yield MoS2 nanosheets to passively Q-switched fiber laser effectively at low pump strengths.

  6. Ultra-broadband nonlinear saturable absorption of high-yield MoS2 nanosheets.

    PubMed

    Wei, Rongfei; Zhang, Hang; Hu, Zhongliang; Qiao, Tian; He, Xin; Guo, Qiangbing; Tian, Xiangling; Chen, Zhi; Qiu, Jianrong

    2016-07-29

    High-yield MoS2 nanosheets with strong nonlinear optical (NLO) responses in a broad near-infrared range were synthesized by a facile hydrothermal method. The observation of saturable absorption, which was excited by the light with photon energy smaller than the gap energy of MoS2, can be attributed to the enhancement of the hybridization between the Mo d-orbital and S p-orbital by the oxygen incorporation into MoS2. High-yield MoS2 nanosheets with high modulation depth and large saturable intensity generated a stable, passively Q-switched fiber laser pulse at 1.56 μm. The high output power of 1.08 mW can be attained under a very low pump power of 30.87 mW. Compared to recently reported passively Q-switched fiber lasers utilizing exfoliated MoS2 nanosheets, the efficiency of the laser for our passive Q-switching operation is larger and reaches 3.50%. This research may extend the understanding on the NLO properties of MoS2 and indicate the feasibility of the high-yield MoS2 nanosheets to passively Q-switched fiber laser effectively at low pump strengths. PMID:27319573

  7. Ultra-broadband nonlinear saturable absorption of high-yield MoS2 nanosheets

    NASA Astrophysics Data System (ADS)

    Wei, Rongfei; Zhang, Hang; Hu, Zhongliang; Qiao, Tian; He, Xin; Guo, Qiangbing; Tian, Xiangling; Chen, Zhi; Qiu, Jianrong

    2016-07-01

    High-yield MoS2 nanosheets with strong nonlinear optical (NLO) responses in a broad near-infrared range were synthesized by a facile hydrothermal method. The observation of saturable absorption, which was excited by the light with photon energy smaller than the gap energy of MoS2, can be attributed to the enhancement of the hybridization between the Mo d-orbital and S p-orbital by the oxygen incorporation into MoS2. High-yield MoS2 nanosheets with high modulation depth and large saturable intensity generated a stable, passively Q-switched fiber laser pulse at 1.56 μm. The high output power of 1.08 mW can be attained under a very low pump power of 30.87 mW. Compared to recently reported passively Q-switched fiber lasers utilizing exfoliated MoS2 nanosheets, the efficiency of the laser for our passive Q-switching operation is larger and reaches 3.50%. This research may extend the understanding on the NLO properties of MoS2 and indicate the feasibility of the high-yield MoS2 nanosheets to passively Q-switched fiber laser effectively at low pump strengths.

  8. Mutant breeding of Serratia marcescens strain for enhancing prodigiosin production and application to textiles.

    PubMed

    Liu, Xiaoxia; Wang, Yujie; Sun, Shiqing; Zhu, Changjun; Xu, Wei; Park, Yongdoo; Zhou, Haimeng

    2013-01-01

    Microwaves have been used as a mutant agent to select mutant strains with high-yield and high-purity pigment. Mass spectrometry and nuclear magnetic resonance spectroscopic techniques were used to elucidate the structures of the pigment. High-performance liquid chromatography was used to measure pigment purity. The analysis of the mutant strain showed that pigment yield increased by 109% and was 98% pure. Prodigiosin in ethanol solution had good stability under ambient temperature and natural indoor light. However, prodigiosin rapidly decomposed under intense sunlight. Prodigiosin is an ecological colorant to dye fabrics, including synthetic and natural fibers. Synthetic fabrics dyed with prodigiosin, such as polyamide and acrylic, have high colorfastness to washing (≥4th grade) and antimicrobial properties (>90%) against Escherichia coli and Staphylococcus aureus. Antimicrobial properties were significantly different between synthetic and natural fabrics. The mutant strain Serratia marcescens jx1-1, with high prodigiosin yield and purity, has promising prospects in food, cosmetic, and textile industries.

  9. Pyrolytic and oxidative syntheses of high-yield carbons from biomass

    NASA Astrophysics Data System (ADS)

    Dai, Xiangfeng

    A pressurized chemical reactor was designed and built per ASME code, which is capable of conducting pyrolysis, activation and gasification processes with an available volume of 7.2 L. It can operate at a pressure up to 3.5 MPa and the system provides a maximum saturated steam flow rate of 2.4 kg/hr at 5.4 MPa. With this reactor, the effects of moisture content, pressure and pretreatment of feedstock on synthesizing charcoal and its yields and properties were examined. Higher moisture content favors charcoal formation and leads to uniform properties of charcoal throughout the bed. However, it demands more external energy. Increase in operation pressure also favors higher yield of charcoal with reduced cook time, but not significantly for pressure above 1 MPa. Woods sorbed with CuSOsb4 and FeSOsb4 result in higher yield and/or better quality as well as short cook time. Charcoals from woods and nutshells have yields around 40% to 65% with a gross calorific value over 30 MJ/kg. A novel process for synthesizing high-yield activated carbon from Macshells was discovered, which involves air activation. Together with equations derived from energy balance, a model describing mass transport and reaction was developed for evaluating roles of diffusion in oxygenating biomass charcoal. Carbonization step is found necessary for subsequent oxygenation and activation in obtaining high-yield activated carbon. Both modeled and experimental results show no influence of mass transport in oxygenation at a temperature below 560 K, and good agreement in temperature rise due to reaction. Results from the kinetics model and the experiments indicate first-order reaction kinetics in oxygenation with respect to oxygen partial pressure. Evaluation on the values of increase in surface area (or iodine number) per carbon loss for activated carbon obtained by this process shows more effective use of carbon compared to the conventional method. Ultimate analyses show high carbon content and low ash

  10. Surface studies and implanted helium measurements following NOVA high-yield DT experiments

    SciTech Connect

    Stoyer, M.A.; Hudson, G.B.

    1997-02-18

    This paper presents the results of three March 6, 1996 direct-drive high-yield DT NOVA experiments and provides `proof-of-principal` results for the quantitative measurement of energetic He ions. Semiconductor quality Si wafers and an amorphous carbon wafer were exposed to NOVA high-yield implosions. Surface damage was sub-micron in general, although the surface ablation was slightly greater for the carbon wafer than for the Si wafers. Melting of a thin ({approx} 0.1{mu}) layer of Si was evident from microscopic investigation. Electron microscopy indicated melted blobs of many different metals (e.g. Al, Au, Ta, Fe alloys, Cu and even Cd) on the surfaces. The yield measured by determining the numbers of atoms of implanted {sup 4}He and {sup 3}He indicate the number of DT fusions to be 9.1({plus_minus}2.3) X 10{sup 12} and DD fusions to be 4.8({plus_minus}1.0) x 10{sup 10}, respectively. The helium DT fusion yield is slightly lower than that of the Cu activation measurement, which was 1.3({plus_minus}0.l) x 10{sup 13} DT fusions.

  11. Designing a high-yielding maize ideotype for a changing climate in Lombardy plain (northern Italy).

    PubMed

    Perego, Alessia; Sanna, Mattia; Giussani, Andrea; Chiodini, Marcello Ermido; Fumagalli, Mattia; Pilu, Salvatore Roberto; Bindi, Marco; Moriondo, Marco; Acutis, Marco

    2014-11-15

    The expected climate change will affect the maize yields in view of air temperature increase and scarce water availability. The application of biophysical models offers the chance to design a drought-resistant ideotype and to assist plant breeders and agronomists in the assessment of its suitability in future scenarios. The aim of the present work was to perform a model-based estimation of the yields of two hybrids, current vs ideotype, under future climate scenarios (2030-2060 and 2070-2100) in Lombardy (northern Italy), testing two options of irrigation (small amount at fixed dates vs optimal water supply), nitrogen (N) fertilization (300 vs 400 kg N ha(-1)), and crop cycle durations (current vs extended). For the designing of the ideotype we set several parameters of the ARMOSA process-based crop model: the root elongation rate and maximum depth, stomatal resistance, four stage-specific crop coefficients for the actual transpiration estimation, and drought tolerance factor. The work findings indicated that the current hybrid ensures good production only with high irrigation amount (245-565 mm y(-1)). With respect to the current hybrid, the ideotype will require less irrigation water (-13%, p<0.01) and it resulted in significantly higher yield under water stress condition (+15%, p<0.01) and optimal water supply (+2%, p<0.05). The elongated cycle has a positive effect on yield under any combination of options. Moreover, higher yields projected for the ideotype implicate more crop residues to be incorporated into the soil, which are positively correlated with the SOC sequestration and negatively with N leaching. The crop N uptake is expected to be adequate in view of higher rate of soil mineralization; the N fertilization rate of 400 kg N ha(-1) will involve significant increasing of grain yield, and it is expected to involve a higher rate of SOC sequestration. PMID:24913890

  12. Differential analysis in Proteome of Space Induced Rice and Soybean Mutants

    NASA Astrophysics Data System (ADS)

    Wang, W.; Lu, B.; Gu, D.; Han, S.; Gao, Y.; Sun, Y.

    To investigate the change trends of proteome induced in space environment we chose 3 Rice mutants 2 Soybean mutants and the seeds which were selected as high yields high tillering rice blast resistance soybean insect pest resistance and wider leaf shape individually after abroad Recoverable Satellite JB-1 for 15 days in 1996 and their corresponding controls Two-dimensional gel electrophoresis 2-D with Coomassie Brilliant Blue staining and PDQuest TM software analysis found that In 6 rice samples 329 pm 35 protein spots were detected in controls whereas 298 pm 37 protein spots detected in mutants representing a 9 decrease 69 pm 27 protein spots were lost in mutants while 37 pm 14 protein spots appeared additionally showing 11 protein spots were lost in mutants 58 protein spots were significantly regulated in mutants with 16 pm 7 up- and 42 pm 18 down-regulated which occupied 5 and 14 of the total average mutants spots separately In 3 soybean leaf samples 263 pm 12 protein spots were detected in controls whereas 255 pm 20 protein spots detected in mutants representing a 3 decrease 49 pm 10 protein spots were lost in mutants while 36 pm 16 protein spots appeared additionally showing 5 protein spots lost in mutants 51 protein spots were significantly regulated in mutants with 25 pm 7 up- and 26 pm 15 down-regulated which occupied 9 8 and 10 2 of the total average mutants spots separately In 3 soybean seed samples 208 pm 41 protein spots were

  13. A shutoff and exonuclease mutant of murine gammaherpesvirus-68 yields infectious virus and causes RNA loss in type I interferon receptor knockout cells.

    PubMed

    Sheridan, Victoria; Polychronopoulos, Louise; Dutia, Bernadette M; Ebrahimi, Bahram

    2014-05-01

    Significant loss of RNA followed by severely reduced cellular protein pool, a phenomenon termed host shutoff, is associated with a number of lytic virus infections and is a critical player in viral pathogenesis. Until recently, viral DNA exonucleases were associated only with processing of viral genomic DNA and its encapsidation. However, recent observations have identified host shutoff and exonuclease function for the highly conserved viral exonucleases in γ-herpesviruses, which include Kaposi's sarcoma-associated herpesvirus, Epstein-Barr virus and the mouse model murine gammaherpesvirus-68, also referred to as MHV-68. In this study, we show that although ablation of the MHV-68 exonuclease ORF37 caused a restrictive phenotype in WT IFN-α/β receptor-positive cells such as NIH 3T3, lack of ORF37 was tolerated in cells lacking the IFN-α/β receptor: the ORF37Stop virus was capable of forming infectious particles and caused loss of mRNA in IFN-α/β receptor knockout cells. Moreover, ORF37Stop virus was able to establish lytic infection in the lungs of mice lacking the IFN-α/β receptor. These observations provide evidence that lytic MHV-68 infection and subsequent loss of mRNA can take place independently of ORF37. Moreover, efficient growth of ORF37Stop virus also identifies a role for this family of viral nucleases in providing a window of opportunity for virus growth by overcoming type I IFN-dependent responses.

  14. Metabolic engineering of biomass for high energy density: oilseed-like triacylglycerol yields from plant leaves.

    PubMed

    Vanhercke, Thomas; El Tahchy, Anna; Liu, Qing; Zhou, Xue-Rong; Shrestha, Pushkar; Divi, Uday K; Ral, Jean-Philippe; Mansour, Maged P; Nichols, Peter D; James, Christopher N; Horn, Patrick J; Chapman, Kent D; Beaudoin, Frederic; Ruiz-López, Noemi; Larkin, Philip J; de Feyter, Robert C; Singh, Surinder P; Petrie, James R

    2014-02-01

    High biomass crops have recently attracted significant attention as an alternative platform for the renewable production of high energy storage lipids such as triacylglycerol (TAG). While TAG typically accumulates in seeds as storage compounds fuelling subsequent germination, levels in vegetative tissues are generally low. Here, we report the accumulation of more than 15% TAG (17.7% total lipids) by dry weight in Nicotiana tabacum (tobacco) leaves by the co-expression of three genes involved in different aspects of TAG production without severely impacting plant development. These yields far exceed the levels found in wild-type leaf tissue as well as previously reported engineered TAG yields in vegetative tissues of Arabidopsis thaliana and N. tabacum. When translated to a high biomass crop, the current levels would translate to an oil yield per hectare that exceeds those of most cultivated oilseed crops. Confocal fluorescence microscopy and mass spectrometry imaging confirmed the accumulation of TAG within leaf mesophyll cells. In addition, we explored the applicability of several existing oil-processing methods using fresh leaf tissue. Our results demonstrate the technical feasibility of a vegetative plant oil production platform and provide for a step change in the bioenergy landscape, opening new prospects for sustainable food, high energy forage, biofuel and biomaterial applications.

  15. Low LET radiolysis escape yields for reducing radicals and H2 in pressurized high temperature water

    NASA Astrophysics Data System (ADS)

    Sterniczuk, Marcin; Yakabuskie, Pamela A.; Wren, J. Clara; Jacob, Jasmine A.; Bartels, David M.

    2016-04-01

    Low Linear Energy Transfer (LET) radiolysis escape yields (G values) are reported for the sum (G(radH)+G(e-)aq) and for G(H2) in subcritical water up to 350 °C. The scavenger system 1-10 mM acetate/0.001 M hydroxide/0.00048 M N2O was used with simultaneous mass spectroscopic detection of H2 and N2 product. Temperature-dependent measurements were carried out with 2.5 MeV electrons from a van de Graaff accelerator, while room temperature calibration measurements were done with a 60Co gamma source. The concentrations and dose range were carefully chosen so that initial spur chemistry is not perturbed and the N2 product yield corresponds to those reducing radicals that escape recombination in pure water. In comparison with a recent review recommendation of Elliot and Bartels (AECL report 153-127160-450-001, 2009), the measured reducing radical yield is seven percent smaller at room temperature but in fairly good agreement above 150 °C. The H2 escape yield is in good agreement throughout the temperature range with several previous studies that used much larger radical scavenging rates. Previous analysis of earlier high temperature measurements of Gesc(radOH) is shown to be flawed, although the actual G values may be nearly correct. The methodology used in the present report greatly reduces the range of possible error and puts the high temperature escape yields for low-LET radiation on a much firmer quantitative foundation than was previously available.

  16. Thin scintillating tiles (with fiber readout and high light-yield) for the OPAL endcaps

    SciTech Connect

    Ball, A.H.

    1998-11-01

    Scintillating tiles with embedded wavelength-shifting fiber readout have recently been installed in the OPAL endcaps to improve trigger performance, time resolution and hermeticity for experiments at LEP II. The design is constrained to provide hermetic coverage of the available area with high single particle efficiency, uniform response and good time resolution, notwithstanding the limited space for the detector and its long readout cables, and despite the strong endcap magnetic field. A high light yield per embedded fiber is required. This paper motivates and describes the design, and demonstrates that the performance meets the required targets. A light yield of 14 photoelectrons/MIP and a time resolution of 3ns have been obtained during 1997 LEP operation. {copyright} {ital 1998 American Institute of Physics.}

  17. Thin scintillating tiles (with fiber readout and high light-yield) for the OPAL endcaps

    SciTech Connect

    Ball, Austin H.

    1998-11-09

    Scintillating tiles with embedded wavelength-shifting fiber readout have recently been installed in the OPAL endcaps to improve trigger performance, time resolution and hermeticity for experiments at LEP II. The design is constrained to provide hermetic coverage of the available area with high single particle efficiency, uniform response and good time resolution, notwithstanding the limited space for the detector and its long readout cables, and despite the strong endcap magnetic field. A high light yield per embedded fiber is required. This paper motivates and describes the design, and demonstrates that the performance meets the required targets. A light yield of 14 photoelectrons/MIP and a time resolution of 3ns have been obtained during 1997 LEP operation.

  18. High-yield isolation and subcellular proteomic characterization of nuclear and subnuclear structures from trypanosomes.

    PubMed

    DeGrasse, Jeffrey A; Chait, Brian T; Field, Mark C; Rout, Michael P

    2008-01-01

    The vast evolutionary distance between the Opisthokonta (animals and yeast) and the excavata (a major group of protists, including Giardia and Trypanosoma) presents a significant challenge to in silico functional genomics and ortholog identification. Subcellular proteomic identification of the constituents of highly enriched organelles can alleviate this problem by both providing localization evidence and yielding a manageably sized proteome for detailed in silico functional assignment. We describe a method for the high-yield isolation of nuclei from the kinetoplastid Trypanosoma brucei. We also describe the subsequent purification of subnuclear compartments, including the nuclear envelope and nucleolus. Finally, using several proteomic strategies, we survey the proteome of a subcellular structure or organelle, using the nuclear pore complex as an example.

  19. A simple, high-yield, apparatus for NEG coating of vacuum beamline elements

    SciTech Connect

    Ron, Guy; Oort, Ron; Lee, Daniel

    2010-12-01

    Non-Evaporable Getter (NEG) materials are extremely useful in vacuum systems for achieving Ultra High Vacuum. Recently, these materials have been used to coat the inner surfaces of vacuum components, acting as an internal, passive, vacuum pump. We have constructed a low cost apparatus, which allows coating of very small diameter vacuum tubes, used as differential pumping stages. Despite the relative ease of construction, we are routinely able to achieve high coating yields. We further describe an improvement to our system, which is able to achieve the same yield, at an even lower complexity by using an easily manufactured permanent magnet arrangement. The designs described are extendible to virtually any combination of length and diameter of the components to be coated.

  20. Bacillus subtilis endospores at high purity and recovery yields: optimization of growth conditions and purification method.

    PubMed

    Tavares, Milene B; Souza, Renata D; Luiz, Wilson B; Cavalcante, Rafael C M; Casaroli, Caroline; Martins, Eduardo G; Ferreira, Rita C C; Ferreira, Luís C S

    2013-03-01

    Bacillus subtilis endospores have applications in different fields including their use as probiotics and antigen delivery vectors. Such specialized applications frequently require highly purified spore preparations. Nonetheless, quantitative data regarding both yields and purity of B. subtilis endospores after application of different growth conditions and purification methods are scarce or poorly reported. In the present study, we conducted several quantitative and qualitative analyses of growth conditions and purification procedures aiming generation of purified B. subtilis spores. Based on two growth media and different incubations conditions, sporulation frequencies up to 74.2 % and spore concentrations up to 7 × 10(9) spores/ml were achieved. Application of a simplified spore isolation method, in which samples were incubated with lysozyme and a detergent, resulted in preparations with highly purified spores at the highest yields. The present study represents, therefore, an important contribution for those working with B. subtilis endospores for different biotechnological purposes.

  1. Fluoro-Julia Olefination as a Mild, High-Yielding Route to α-Fluoro Acrylonitriles†

    PubMed Central

    del Solar, Maria; Ghosh, Arun K.; Zajc, Barbara

    2009-01-01

    Synthesis of a novel, stable reagent (1,3-benzothiazol-2-ylsulfonyl)fluoroacetonitrile from readily available ethyl α-(1,3-benzothiazol-2-ylsulfanyl)- α-fluoroacetate is reported. Aldehydes undergo condensations with (1,3-benzothiazol-2-ylsulfonyl)fluoroacetonitrile in the presence of DBU leading to α-fluoro acrylonitriles in high yields and with good Z-stereoselectivity. Lowering of reaction temperature increases the Z selectivity. PMID:18841918

  2. Development of High Yield Feedstocks and Biomass Conversion Technology for Renewable Energy

    SciTech Connect

    Hashimoto, Andrew G.; Crow, Susan; DeBeryshe, Barbara; Ha, Richard; Jakeway, Lee; Khanal, Samir; Nakahata, Mae; Ogoshi, Richard; Shimizu, Erik; Stern, Ivette; Turano, Brian; Turn, Scott; Yanagida, John

    2015-04-09

    This project had two main goals. The first goal was to evaluate several high yielding tropical perennial grasses as feedstock for biofuel production, and to characterize the feedstock for compatible biofuel production systems. The second goal was to assess the integration of renewable energy systems for Hawaii. The project focused on high-yield grasses (napiergrass, energycane, sweet sorghum, and sugarcane). Field plots were established to evaluate the effects of elevation (30, 300 and 900 meters above sea level) and irrigation (50%, 75% and 100% of sugarcane plantation practice) on energy crop yields and input. The test plots were extensive monitored including: hydrologic studies to measure crop water use and losses through seepage and evapotranspiration; changes in soil carbon stock; greenhouse gas flux (CO2, CH4, and N2O) from the soil surface; and root morphology, biomass, and turnover. Results showed significant effects of environment on crop yields. In general, crop yields decrease as the elevation increased, being more pronounced for sweet sorghum and energycane than napiergrass. Also energy crop yields were higher with increased irrigation levels, being most pronounced with energycane and less so with sweet sorghum. Daylight length greatly affected sweet sorghum growth and yields. One of the energy crops (napiergrass) was harvested at different ages (2, 4, 6, and 8 months) to assess the changes in feedstock characteristics with age and potential to generate co-products. Although there was greater potential for co-products from younger feedstock, the increased production was not sufficient to offset the additional cost of harvesting multiple times per year. The feedstocks were also characterized to assess their compatibility with biochemical and thermochemical conversion processes. The project objectives are being continued through additional support from the Office of Naval Research, and the Biomass Research and Development

  3. Epidemiology and impact of Fasciola hepatica exposure in high-yielding dairy herds

    PubMed Central

    Howell, Alison; Baylis, Matthew; Smith, Rob; Pinchbeck, Gina; Williams, Diana

    2015-01-01

    The liver fluke Fasciola hepatica is a trematode parasite with a worldwide distribution and is the cause of important production losses in the dairy industry. The aim of this observational study was to assess the prevalence of exposure to F. hepatica in a group of high yielding dairy herds, to determine the risk factors and investigate their associations with production and fertility parameters. Bulk milk tank samples from 606 herds that supply a single retailer with liquid milk were tested with an antibody ELISA for F. hepatica. Multivariable linear regression was used to investigate the effect of farm management and environmental risk factors on F. hepatica exposure. Higher rainfall, grazing boggy pasture, presence of beef cattle on farm, access to a stream or pond and smaller herd size were associated with an increased risk of exposure. Univariable regression was used to look for associations between fluke exposure and production-related variables including milk yield, composition, somatic cell count and calving index. Although causation cannot be assumed, a significant (p < 0.001) negative association was seen between F. hepatica exposure and estimated milk yield at the herd level, representing a 15% decrease in yield for an increase in F. hepatica exposure from the 25th to the 75th percentile. This remained significant when fertility, farm management and environmental factors were controlled for. No associations were found between F. hepatica exposure and any of the other production, disease or fertility variables. PMID:26093971

  4. A novel medium devoid of ruminant peptone for high yield growth of Mycoplasma ovipneumoniae.

    PubMed

    Patel, Hiren; Mackintosh, David; Ayling, Roger D; Nicholas, Robin A J; Fielder, Mark D

    2008-03-18

    Mycoplasma ovipneumoniae is considered an emerging veterinary pathogen causing pneumonia in sheep and goats worldwide. Currently it has not been possible to define a growth medium that yields the maximum growth of M. ovipneumoniae within a short incubation period. Growth yields of M. ovipneumoniae in Eaton's medium are variable and not as consistently high as those seen with other Mycoplasma spp. This study investigated the ability of different M. ovipneumoniae field strains to grow in various media formulations, where PPLO broth was replaced by a vegetable protein source, and comparisons were made in terms of strain viability in Eaton's medium. Studies were also conducted to determine the optimal carbohydrate source for use in the M. ovipneumoniae medium. Generally, it was found that different strains showed good growth in all media tested, with growth yields at 24h in TSB-1 medium higher than those observed with Eaton's medium. Growth yields reached 10(8) to 10(9)cfu ml(-1) within 24h for particular field strains, with all strains achieving this growth level within 48-72h.

  5. Epidemiology and impact of Fasciola hepatica exposure in high-yielding dairy herds.

    PubMed

    Howell, Alison; Baylis, Matthew; Smith, Rob; Pinchbeck, Gina; Williams, Diana

    2015-09-01

    The liver fluke Fasciola hepatica is a trematode parasite with a worldwide distribution and is the cause of important production losses in the dairy industry. The aim of this observational study was to assess the prevalence of exposure to F. hepatica in a group of high yielding dairy herds, to determine the risk factors and investigate their associations with production and fertility parameters. Bulk milk tank samples from 606 herds that supply a single retailer with liquid milk were tested with an antibody ELISA for F. hepatica. Multivariable linear regression was used to investigate the effect of farm management and environmental risk factors on F. hepatica exposure. Higher rainfall, grazing boggy pasture, presence of beef cattle on farm, access to a stream or pond and smaller herd size were associated with an increased risk of exposure. Univariable regression was used to look for associations between fluke exposure and production-related variables including milk yield, composition, somatic cell count and calving index. Although causation cannot be assumed, a significant (p<0.001) negative association was seen between F. hepatica exposure and estimated milk yield at the herd level, representing a 15% decrease in yield for an increase in F. hepatica exposure from the 25th to the 75th percentile. This remained significant when fertility, farm management and environmental factors were controlled for. No associations were found between F. hepatica exposure and any of the other production, disease or fertility variables.

  6. Construction high-yield candidate influenza vaccine viruses in Vero cells by reassortment.

    PubMed

    Yu, Wei; Yang, Fan; Yang, Jinghui; Ma, Lei; Cun, Yina; Song, Shaohui; Liao, Guoyang

    2016-11-01

    Usage of influenza vaccine is the best choice measure for preventing and conclusion of influenza virus infection. Although it has been used of chicken embryo to produce influenza vaccine, following with WHO recommended vaccine strain, there were uncontrollable factors and its deficiencies, specially, during an influenza pandemic in the world. The Vero cells are used for vaccine production of a few strains including influenza virus, because of its homology with human, recommended by WHO. However, as known most of the influenza viruses strains could not culture by Vero cells. It was used two high-yield influenza viruses adapted in Vero cells as donor viruses, such as A/Yunnan/1/2005Va (H3N2) and B/Yunnan/2/2005Va (B), to construct high-yield wild influenza virus in Vero cells under antibody selection pressure. After reassortment and passages, it obtained the new Vaccine strains with A/Tianjin/15/2009Va (H1N1), A/Fujian/196/2009Va (H3N2) and B/Chongqing/1384/2010Va (B), which was not only completely keeping their original antigenic (HA and NA), but also grown well in Vero cells with high-yield. All results of gene analysis and HA, HI shown that this reassortment method could be used to find new direction to product the influenza vaccine. J. Med. Virol. 88:1914-1921, 2016. © 2016 Wiley Periodicals, Inc. PMID:27101353

  7. Experimental quantification of cavitation yield revisited: focus on high frequency ultrasound reactors.

    PubMed

    Kirpalani, D M; McQuinn, K J

    2006-01-01

    Acoustic cavitation plays an important role in enhancing the reaction rate of chemical processes in sonochemical systems. However, quantification of cavitation intensity in sonochemical systems is generally limited to low frequency systems. In this study, an empirical determination of cavitation yield in high frequency ultrasound systems was performed by measuring the amount of iodine liberated from the oxidation of potassium iodide (KI) solution at 1.7 and 2.4 MHz. Experiments for determining cavitation were carried out at various solute (KI) concentrations under constant temperature, obtained by direct cooling of the solution and variable temperature conditions, in the absence of external cooling. Cavitation yield measurements, reported in this work, extend previously reported results and lend credence to the two step reaction pathway in high frequency systems. Additionally, the concentration of KI and temperature affect the cavitation yield of a system such that the iodine production is proportional to both conditions. It is proposed that direct cooling of sonicated KI solution may be advantageous for optimization of cavitation intensity in high frequency sonochemical reactors.

  8. Variable sensitivity of US maize yield to high temperatures across developmental stages

    NASA Astrophysics Data System (ADS)

    Butler, E. E.; Huybers, P. J.

    2013-12-01

    The sensitivity of maize to high temperatures has been widely demonstrated. Furthermore, field work has indicated that reproductive development stages are particularly sensitive to stress, but this relationship has not been quantified across a wide geographic region. Here, the relationship between maize yield and temperature variations is examined as a function of developmental stage. US state-level data from the National Agriculture Statistics Service provide dates for six growing stages: planting, silking, doughing, dented, mature, and harvested. Temperatures that correspond to each developmental stage are then inferred from a network of weather station observations interpolated to the county level, and a multiple linear regression technique is employed to estimate the sensitivity of county yield outcomes to variations in growing-degree days and an analogous measure of high temperatures referred to as killing-degree days. Uncertainties in the transition times between county-level growth stages are accounted for. Results indicate that the silking and dented stages are generally the most sensitive to killing degree days, with silking the most sensitive stage in the US South and dented the most sensitive in the US North. These variable patterns of sensitivity aid in interpreting which weather events are of greatest significance to maize yields and provide some insight into how shifts in planting time or changes in developmental timing would influence the risks associated with exposure to high temperatures.

  9. Isolation of Hofbauer Cells from Human Term Placentas with High Yield and Purity

    PubMed Central

    Tang, Zhonghua; Tadesse, Serkalem; Norwitz, Errol; Mor, Gil; Abrahams, Vikki M.; Guller, Seth

    2011-01-01

    Problem Placental villus macrophages (i.e. Hofbauer cells, HBCs) were identified more than 100 years ago. Alterations in their numbers and characteristics are associated with several complications of pregnancy. Although HBCs have previously been isolated and cultured, there is no consensus methodology to obtain these cells with high yield and purity for in vitro studies. Method of Study HBCs were isolated from human term placentas using protocols in which cytotrophoblasts (CTs) and fibroblasts (FIBs), other major villous cell types, were isolated in parallel. Enzymatic digestion, Percoll gradients, and immunoselection were used to isolate the three cell types. Purity was assessed by morphology, flow cytometry, and in phagocytosis assays. Results HBCs were isolated with 98–99% purity and a yield of 130–200 ×106 cells/80 to 100 g of tissue. HBCs exhibited a pleiomorphic and vacuolated appearance for at least 5 days in culture medium with and without serum. High levels of phagocytosis in HBCs, but not in CTs, or FIBs, confirmed macrophage function in HBCs. Phagocytotic activity was maintained across several days in culture. Conclusion HBCs were isolated from term placenta with high yield and purity using protocols in which CTs and FIBs were also obtained. This methodology will foster future studies which examine the role of HBCs in regulating villus function. PMID:21545365

  10. Synthesis of Luminescent Graphene Quantum Dots with High Quantum Yield and Their Toxicity Study

    PubMed Central

    Jiang, Dan; Chen, Yunping; Li, Na; Li, Wen; Wang, Zhenguo; Zhu, Jingli; Zhang, Hong; Liu, Bin; Xu, Shan

    2015-01-01

    High fluorescence quantum yield graphene quantum dots (GQDs) have showed up as a new generation for bioimaging. In this work, luminescent GQDs were prepared by an ameliorative photo-Fenton reaction and a subsequent hydrothermal process using graphene oxide sheets as the precursor. The as-prepared GQDs were nanomaterials with size ranging from 2.3 to 6.4 nm and emitted intense green luminescence in water. The fluorescence quantum yield was as high as 24.6% (excited at 340 nm) and the fluorescence was strongest at pH 7. Moreover, the influences of low-concentration (12.5, 25 μg/mL) GQDs on the morphology, viability, membrane integrity, internal cellular reactive oxygen species level and mortality of HeLa cells were relatively weak, and the in vitro imaging demonstrated GQDs were mainly in the cytoplasm region. More strikingly, zebrafish embryos were co-cultured with GQDs for in vivo imaging, and the results of heart rate test showed the intake of small amounts of GQDs brought little harm to the cardiovascular of zebrafish. GQDs with high quantum yield and strong photoluminescence show good biocompatibility, thus they show good promising for cell imaging, biolabeling and other biomedical applications. PMID:26709828

  11. The variation of the yield stress of Ti alloys with strain rate at high temperatures

    NASA Astrophysics Data System (ADS)

    Rosen, R. S.; Paddon, S. P.; Kassner, M. E.

    1999-06-01

    This study extended investigation on the elevated-temperature yield-strength dependence of beta-phase titanium alloys on strain rate and temperature. Yield stresses were found to increase substantially with increasing strain rate at elevated temperatures due to the high strain-rate sensitivity of titanium at high temperatures. Above 1000 °C, the strain-rate sensitivities were found to increase substantially with increasing temperature and/or decreasing strain rate. The six alloys examined were TIMETAL 21S, Ti-15-3, Ti-6-4, Ti-13-11-3, Beta C, and Beta III. There was particular interest in determining the strain-rate sensitivity of these alloys through strain-rate change tests above 1000 °C. The yield stresses of all the titanium alloys at temperatures above 1093 °C were less than 1% of their ambient temperature values. Strain hardening was negligible in the alloys tested at these high temperatures. Extended tensile ductilities of 100 to 200% were observed due to the pronounced strain-rate sensitivity. The rate controlling mechanism for plasticity, based on activation energy and the strain-rate sensitivity measurements, is discussed.

  12. The variation of the yield stress of Ti alloys with strain rate at high temperatures

    SciTech Connect

    Rosen, R.S.; Paddon, S.P.; Kassner, M.E.

    1999-06-01

    This study extended investigation on the elevated-temperature yield-strength dependence of beta-phase titanium alloys on strain rate and temperature. Yield stresses were found to increase substantially with increasing strain rate at elevated temperatures due to the high strain-rate sensitivity of titanium at high temperatures. Above 1000 C, the strain-rate sensitivities were found to increase substantially with increasing temperature and/or decreasing strain rate. The six alloys examined were TIMETAL 21S, Ti-15-3-, Ti-6-4, Ti-13-11-3, Beta C, and Beta III. There was particular interest in determining the strain-rate sensitivity of these alloys through strain-rate change tests above 1000 C. The yield stresses of all the titanium alloys at temperatures above 1093 C were less than 1% of their ambient temperature values. strain hardening was negligible in the alloys tested at these high temperatures. Extended tensile ductilities of 100 to 200% were observed due to the pronounced strain-rare sensitivity. The rate controlling mechanism for plasticity, based on activation energy and the strain-rate sensitivity measurements, is discussed.

  13. Predawn respiration rates during flowering are highly predictive of yield response in Gossypium hirsutum when yield variability is water-induced.

    PubMed

    Snider, John L; Chastain, Daryl R; Meeks, Calvin D; Collins, Guy D; Sorensen, Ronald B; Byrd, Seth A; Perry, Calvin D

    2015-07-01

    Respiratory carbon evolution by leaves under abiotic stress is implicated as a major limitation to crop productivity; however, respiration rates of fully expanded leaves are positively associated with plant growth rates. Given the substantial sensitivity of plant growth to drought, it was hypothesized that predawn respiration rates (RPD) would be (1) more sensitive to drought than photosynthetic processes and (2) highly predictive of water-induced yield variability in Gossypium hirsutum. Two studies (at Tifton and Camilla Georgia) addressed these hypotheses. At Tifton, drought was imposed beginning at the onset of flowering (first flower) and continuing for three weeks (peak bloom) followed by a recovery period, and predawn water potential (ΨPD), RPD, net photosynthesis (AN) and maximum quantum yield of photosystem II (Fv/Fm) were measured throughout the study period. At Camilla, plants were exposed to five different irrigation regimes throughout the growing season, and average ΨPD and RPD were determined between first flower and peak bloom for all treatments. For both sites, fiber yield was assessed at crop maturity. The relationships between ΨPD, RPD and yield were assessed via non-linear regression. It was concluded for field-grown G. hirsutum that (1) RPD is exceptionally sensitive to progressive drought (more so than AN or Fv/Fm) and (2) average RPD from first flower to peak bloom is highly predictive of water-induced yield variability. PMID:26125121

  14. Economics of fertility in high-yielding dairy cows on confined TMR systems.

    PubMed

    Cabrera, V E

    2014-05-01

    The objective of this review paper was to summarise the latest findings in dairy cattle reproductive economics with an emphasis on high yielding, confined total mixed ration systems. The economic gain increases as the reproductive efficiency improves. These increments follow the law of diminishing returns, but are still positive even at high reproductive performance. Reproductive improvement results in higher milk productivity and, therefore, higher milk income over feed cost, more calf sales and lower culling and breeding expenses. Most high-yielding herds in the United States use a combination of timed artificial insemination (TAI) and oestrous detection (OD) reproductive programme. The ratio of achievable pregnancies between OD and TAI determines the economic value difference between both and their combinations. Nonetheless, complex interactions between reproductive programme, herd relative milk yield, and type of reproductive programme are reported. For example, higher herd relative milk yield would favour programme relying more on TAI. In addition, improved reproductive efficiency produces extra replacements. The availability of additional replacements could allow more aggressive culling policies (e.g. less services for non-pregnant cows) to balance on-farm supply and demand of replacements. Balancing heifer replacement availability in an efficient reproductive programme brings additional economic benefits. New technologies such as the use of earlier chemical tests for pregnancy diagnosis could be economically effective depending on the goals and characteristics of the farm. Opportunities for individual cow reproductive management within defined reproductive programme exist. These decisions would be based on economic metrics derived from the value of a cow such as the value of a new pregnancy, the cost of a pregnancy loss, or the cost of an extra day open. PMID:24679357

  15. Economics of fertility in high-yielding dairy cows on confined TMR systems.

    PubMed

    Cabrera, V E

    2014-05-01

    The objective of this review paper was to summarise the latest findings in dairy cattle reproductive economics with an emphasis on high yielding, confined total mixed ration systems. The economic gain increases as the reproductive efficiency improves. These increments follow the law of diminishing returns, but are still positive even at high reproductive performance. Reproductive improvement results in higher milk productivity and, therefore, higher milk income over feed cost, more calf sales and lower culling and breeding expenses. Most high-yielding herds in the United States use a combination of timed artificial insemination (TAI) and oestrous detection (OD) reproductive programme. The ratio of achievable pregnancies between OD and TAI determines the economic value difference between both and their combinations. Nonetheless, complex interactions between reproductive programme, herd relative milk yield, and type of reproductive programme are reported. For example, higher herd relative milk yield would favour programme relying more on TAI. In addition, improved reproductive efficiency produces extra replacements. The availability of additional replacements could allow more aggressive culling policies (e.g. less services for non-pregnant cows) to balance on-farm supply and demand of replacements. Balancing heifer replacement availability in an efficient reproductive programme brings additional economic benefits. New technologies such as the use of earlier chemical tests for pregnancy diagnosis could be economically effective depending on the goals and characteristics of the farm. Opportunities for individual cow reproductive management within defined reproductive programme exist. These decisions would be based on economic metrics derived from the value of a cow such as the value of a new pregnancy, the cost of a pregnancy loss, or the cost of an extra day open.

  16. Method for reproducibly preparing a low-melting high-carbon yield precursor

    DOEpatents

    Smith, Wesley E.; Napier, Jr., Bradley

    1978-01-01

    The present invention is directed to a method for preparing a reproducible synthetic carbon precursor by the autoclave polymerization of indene (C.sub.9 H.sub.8) at a temperature in the range of 470.degree.-485.degree. C, and at a pressure in the range of about 1000 to about 4300 psi. Volatiles in the resulting liquid indene polymer are removed by vacuum outgassing to form a solid carbon precursor characterized by having a relatively low melting temperature, high-carbon yield, and high reproducibility which provide for the fabrication of carbon and graphite composites having strict requirements for reproducible properties.

  17. High-Yield Hydrogen Production from Starch and Water by a Synthetic Enzymatic Pathway

    PubMed Central

    Zhang, Y.-H. Percival; Evans, Barbara R.; Mielenz, Jonathan R.; Hopkins, Robert C.; Adams, Michael W.W.

    2007-01-01

    Background The future hydrogen economy offers a compelling energy vision, but there are four main obstacles: hydrogen production, storage, and distribution, as well as fuel cells. Hydrogen production from inexpensive abundant renewable biomass can produce cheaper hydrogen, decrease reliance on fossil fuels, and achieve zero net greenhouse gas emissions, but current chemical and biological means suffer from low hydrogen yields and/or severe reaction conditions. Methodology/Principal Findings Here we demonstrate a synthetic enzymatic pathway consisting of 13 enzymes for producing hydrogen from starch and water. The stoichiometric reaction is C6H10O5 (l)+7 H2O (l)→12 H2 (g)+6 CO2 (g). The overall process is spontaneous and unidirectional because of a negative Gibbs free energy and separation of the gaseous products with the aqueous reactants. Conclusions Enzymatic hydrogen production from starch and water mediated by 13 enzymes occurred at 30°C as expected, and the hydrogen yields were much higher than the theoretical limit (4 H2/glucose) of anaerobic fermentations. Significance The unique features, such as mild reaction conditions (30°C and atmospheric pressure), high hydrogen yields, likely low production costs ($∼2/kg H2), and a high energy-density carrier starch (14.8 H2-based mass%), provide great potential for mobile applications. With technology improvements and integration with fuel cells, this technology also solves the challenges associated with hydrogen storage, distribution, and infrastructure in the hydrogen economy. PMID:17520015

  18. Enhanced electron yield from laser-driven wakefield acceleration in high-Z gas jets.

    PubMed

    Mirzaie, Mohammad; Hafz, Nasr A M; Li, Song; Liu, Feng; He, Fei; Cheng, Ya; Zhang, Jie

    2015-10-01

    An investigation of the electron beam yield (charge) form helium, nitrogen, and neon gas jet plasmas in a typical laser-plasma wakefield acceleration experiment is carried out. The charge measurement is made by imaging the electron beam intensity profile on a fluorescent screen into a charge coupled device which was cross-calibrated with an integrated current transformer. The dependence of electron beam charge on the laser and plasma conditions for the aforementioned gases are studied. We found that laser-driven wakefield acceleration in low Z-gas jet targets usually generates high-quality and well-collimated electron beams with modest yields at the level of 10-100 pC. On the other hand, filamentary electron beams which are observed from high-Z gases at higher densities reached much higher yields. Evidences for cluster formation were clearly observed in the nitrogen gas jet target, where we received the highest electron beam charge of ∼1.7 nC. Those intense electron beams will be beneficial for the applications on the generation of bright X-rays, gamma rays radiations, and energetic positrons via the bremsstrahlung or inverse-scattering processes.

  19. Enhanced electron yield from laser-driven wakefield acceleration in high-Z gas jets

    NASA Astrophysics Data System (ADS)

    Mirzaie, Mohammad; Hafz, Nasr A. M.; Li, Song; Liu, Feng; He, Fei; Cheng, Ya; Zhang, Jie

    2015-10-01

    An investigation of the electron beam yield (charge) form helium, nitrogen, and neon gas jet plasmas in a typical laser-plasma wakefield acceleration experiment is carried out. The charge measurement is made by imaging the electron beam intensity profile on a fluorescent screen into a charge coupled device which was cross-calibrated with an integrated current transformer. The dependence of electron beam charge on the laser and plasma conditions for the aforementioned gases are studied. We found that laser-driven wakefield acceleration in low Z-gas jet targets usually generates high-quality and well-collimated electron beams with modest yields at the level of 10-100 pC. On the other hand, filamentary electron beams which are observed from high-Z gases at higher densities reached much higher yields. Evidences for cluster formation were clearly observed in the nitrogen gas jet target, where we received the highest electron beam charge of ˜1.7 nC. Those intense electron beams will be beneficial for the applications on the generation of bright X-rays, gamma rays radiations, and energetic positrons via the bremsstrahlung or inverse-scattering processes.

  20. Enhanced electron yield from laser-driven wakefield acceleration in high-Z gas jets

    SciTech Connect

    Mirzaie, Mohammad; Hafz, Nasr A. M. Li, Song; Liu, Feng; Zhang, Jie; He, Fei; Cheng, Ya

    2015-10-15

    An investigation of the electron beam yield (charge) form helium, nitrogen, and neon gas jet plasmas in a typical laser-plasma wakefield acceleration experiment is carried out. The charge measurement is made by imaging the electron beam intensity profile on a fluorescent screen into a charge coupled device which was cross-calibrated with an integrated current transformer. The dependence of electron beam charge on the laser and plasma conditions for the aforementioned gases are studied. We found that laser-driven wakefield acceleration in low Z-gas jet targets usually generates high-quality and well-collimated electron beams with modest yields at the level of 10-100 pC. On the other hand, filamentary electron beams which are observed from high-Z gases at higher densities reached much higher yields. Evidences for cluster formation were clearly observed in the nitrogen gas jet target, where we received the highest electron beam charge of ∼1.7 nC. Those intense electron beams will be beneficial for the applications on the generation of bright X-rays, gamma rays radiations, and energetic positrons via the bremsstrahlung or inverse-scattering processes.

  1. Enhanced electron yield from laser-driven wakefield acceleration in high-Z gas jets.

    PubMed

    Mirzaie, Mohammad; Hafz, Nasr A M; Li, Song; Liu, Feng; He, Fei; Cheng, Ya; Zhang, Jie

    2015-10-01

    An investigation of the electron beam yield (charge) form helium, nitrogen, and neon gas jet plasmas in a typical laser-plasma wakefield acceleration experiment is carried out. The charge measurement is made by imaging the electron beam intensity profile on a fluorescent screen into a charge coupled device which was cross-calibrated with an integrated current transformer. The dependence of electron beam charge on the laser and plasma conditions for the aforementioned gases are studied. We found that laser-driven wakefield acceleration in low Z-gas jet targets usually generates high-quality and well-collimated electron beams with modest yields at the level of 10-100 pC. On the other hand, filamentary electron beams which are observed from high-Z gases at higher densities reached much higher yields. Evidences for cluster formation were clearly observed in the nitrogen gas jet target, where we received the highest electron beam charge of ∼1.7 nC. Those intense electron beams will be beneficial for the applications on the generation of bright X-rays, gamma rays radiations, and energetic positrons via the bremsstrahlung or inverse-scattering processes. PMID:26520950

  2. Picosecond Lifetimes with High Quantum Yields from Single-Photon-Emitting Colloidal Nanostructures at Room Temperature.

    PubMed

    Bidault, Sébastien; Devilez, Alexis; Maillard, Vincent; Lermusiaux, Laurent; Guigner, Jean-Michel; Bonod, Nicolas; Wenger, Jérôme

    2016-04-26

    Minimizing the luminescence lifetime while maintaining a high emission quantum yield is paramount in optimizing the excitation cross-section, radiative decay rate, and brightness of quantum solid-state light sources, particularly at room temperature, where nonradiative processes can dominate. We demonstrate here that DNA-templated 60 and 80 nm diameter gold nanoparticle dimers, featuring one fluorescent molecule, provide single-photon emission with lifetimes that can fall below 10 ps and typical quantum yields in a 45-70% range. Since these colloidal nanostructures are obtained as a purified aqueous suspension, fluorescence spectroscopy can be performed on both fixed and freely diffusing nanostructures to quantitatively estimate the distributions of decay rate and fluorescence intensity enhancements. These data are in excellent agreement with theoretical calculations and demonstrate that millions of bright fluorescent nanostructures, with radiative lifetimes below 100 ps, can be produced in parallel. PMID:26972678

  3. Reductive burning of high-yield spent pulping liquors by the addition of pulverized coal

    SciTech Connect

    Sell, N.J.; Norman, J.C. )

    1992-10-01

    This paper reports on the reductive burning of high-yield spent pulping liquors which can be accomplished by the addition of pulverized coal to increase the heat content and generate the proper reducing conditions. Samples from a 78%-yield sodium bisulfite pulping process employing a hardwood furnish were mixed with 10-50% pulveriized coal and burned at 950[degrees]C under reducing conditions in a box furnace. Even in these uncontrolled combustion conditions 76. 5% of the sulfur found in the soluble portion of the smelt was converted from lignousulfonates to useful sulfide ion. For the remainder of the sulfur, analyses determined it to be 19. 5% as sulfite ion, 3. 1% as thiosulfate ion, and 0.9% as sulfate ion.

  4. Shock-Ignited High Gain/Yield Targets for the National Ignition Facility

    NASA Astrophysics Data System (ADS)

    Perkins, L. J.; Lafortune, K. N.; Bedrosiian, P.; Tabak, M.; Miles, A.; Dixit, S.; Betti, R.; Anderson, K.; Zhou, C.

    2006-10-01

    Shock-ignition, a new concept for ICF ignition [C.Zhou, R.Betti Bull APS, v50, 2005], is being studied as a future option for efficiently achieving high gains in large laser facilities such as NIF. Accordingly, this offers the potential for testing: (1)High yield (up to 200MJ), reactor-relevant targets for inertial fusion energy (2)High fusion yield targets for DOE NNSA stockpile application (3)Targets with appreciable gain at low laser drive energies (gains of 10's at 150kJ) (4)Ignition of simple, non-cryo (room temperature) single shell gas targets at (unity gain). By contrast to conventional hotspot ignition, we separate the assembly and ignition phases by initially imploding a massive cryogenic shell on a low adiabat (alpha 0.7) at low velocity (less than 2e7cm/s) using a direct drive pulse of modest total energy. The assembled fuel is then separately ignited by a strong, spherically convergent shock driven by a high intensity spike at the end of the pulse and timed to reach the center as the main fuel is stagnating and starting to rebound. Like fast ignition, shock ignition can achieve high gains with low drive energy, but has the advantages of requiring only a single laser with less demanding timing and spatial focusing requirements.

  5. High prevalence of dhfr triple mutant and correlation with high rates of sulphadoxine-pyrimethamine treatment failures in vivo in Gabonese children

    PubMed Central

    2011-01-01

    Background Drug resistance contributes to the global malaria burden. Plasmodium falciparum dihydrofolate reductase (dhfr) and dihydropteroate synthase (dhps) polymorphisms confer resistance to sulphadoxine-pyrimethamine (SP). Methods The study assessed the frequency of SP resistance-conferring polymorphisms in Plasmodium falciparum-positive samples from two clinical studies in Lambaréné. Their role on treatment responses and transmission potential was studied in an efficacy open-label clinical trial with a 28-day follow-up in 29 children under five with uncomplicated malaria. Results SP was well tolerated by all subjects in vivo. Three subjects were excluded from per-protocol analysis. PCR-corrected, 12/26 (46%) achieved an adequate clinical and parasitological response, 13/26 (50%) were late parasitological failures, while 1/26 (4%) had an early treatment failure, resulting in early trial discontinuation. Of 106 isolates, 98 (92%) carried the triple mutant dhfr haplotype. Three point mutations were found in dhps in a variety of haplotypic configurations. The 437G + 540E double mutant allele was found for the first time in Gabon. Conclusions There is a high prevalence of dhfr triple mutant with some dhps point mutations in Gabon, in line with treatment failures observed, and molecular markers of SP resistance should be closely monitored. Trial Registration ClinicalTrials.gov: NCT00453856 PMID:21569596

  6. High-rate, high-yield production of methanol by ammonia-oxidizing bacteria.

    PubMed

    Taher, Edris; Chandran, Kartik

    2013-04-01

    The overall goal of this study was to develop an appropriate biological process for achieving autotrophic conversion of methane (CH(4)) to methanol (CH3OH). In this study, we employed ammonia-oxidizing bacteria (AOB) to selectively and partially oxidize CH(4) to CH(3)OH. In fed-batch reactors using mixed nitrifying enrichment cultures from a continuous bioreactor, up to 59.89 ± 1.12 mg COD/L of CH(3)OH was produced within an incubation time of 7 h, which is approximately ten times the yield obtained previously using pure cultures of Nitrosomonas europaea. The maximum specific rate of CH(4) to CH(3)OH conversion obtained during this study was 0.82 mg CH(3)OH COD/mg AOB biomass COD-d, which is 1.5 times the highest value reported with pure cultures. Notwithstanding these positive results, CH(4) oxidation to CH(3)OH by AOB was inhibited by NH(3) (the primary substrate for the oxidative enzyme, ammonia monooxygenase, AMO) as well as the product, CH(3)OH, itself. Further, oxidation of CH(4) to CH(3)OH by AOB was also limited by reducing equivalents supply, which could be overcome by externally supplying hydroxylamine (NH(2)OH) as an electron donor. Therefore, a potential optimum design for promoting CH(4) to CH(3)OH oxidation by AOB could involve supplying NH(3) (needed to maintain AMO activity) uncoupled from the supply of NH(2)OH and CH(4). Partial oxidation of CH(4)-containing gases to CH3OH by AOB represents an attractive platform for the conversion of a gaseous mixture to an aqueous compound, which could be used as a commodity chemical. Alternately, the nitrate and CH(3) OH thus produced could be channeled to a downstream anoxic zone in a biological nitrogen removal process to effect nitrate reduction to N(2), using an internally produced organic electron donor.

  7. High Yield Chemical Vapor Deposition Growth of High Quality Large-Area AB Stacked Bilayer Graphene

    PubMed Central

    Liu, Lixin; Zhou, Hailong; Cheng, Rui; Yu, Woo Jong; Liu, Yuan; Chen, Yu; Shaw, Jonathan; Zhong, Xing; Huang, Yu; Duan, Xiangfeng

    2012-01-01

    Bernal stacked (AB stacked) bilayer graphene is of significant interest for functional electronic and photonic devices due to the feasibility to continuously tune its band gap with a vertical electrical field. Mechanical exfoliation can be used to produce AB stacked bilayer graphene flakes but typically with the sizes limited to a few micrometers. Chemical vapor deposition (CVD) has been recently explored for the synthesis of bilayer graphene but usually with limited coverage and a mixture of AB and randomly stacked structures. Herein we report a rational approach to produce large-area high quality AB stacked bilayer graphene. We show that the self-limiting effect of graphene growth on Cu foil can be broken by using a high H2/CH4 ratio in a low pressure CVD process to enable the continued growth of bilayer graphene. A high temperature and low pressure nucleation step is found to be critical for the formation of bilayer graphene nuclei with high AB stacking ratio. A rational design of a two-step CVD process is developed for the growth of bilayer graphene with high AB stacking ratio (up to 90 %) and high coverage (up to 99 %). The electrical transport studies demonstrated that devices made of the as-grown bilayer graphene exhibit typical characteristics of AB stacked bilayer graphene with the highest carrier mobility exceeding 4,000 cm2/V·s at room temperature, comparable to that of the exfoliated bilayer graphene. PMID:22906199

  8. Future consequences of decreasing marginal production efficiency in the high-yielding dairy cow.

    PubMed

    Moallem, U

    2016-04-01

    The objectives were to examine the gross and marginal production efficiencies in high-yielding dairy cows and the future consequences on dairy industry profitability. Data from 2 experiments were used in across-treatments analysis (n=82 mid-lactation multiparous Israeli-Holstein dairy cows). Milk yields, body weights (BW), and dry matter intakes (DMI) were recorded daily. In both experiments, cows were fed a diet containing 16.5 to 16.6% crude protein and net energy for lactation (NEL) at 1.61 Mcal/kg of dry matter (DM). The means of milk yield, BW, DMI, NEL intake, and energy required for maintenance were calculated individually over the whole study, and used to calculate gross and marginal efficiencies. Data were analyzed in 2 ways: (1) simple correlation between variables; and (2) cows were divided into 3 subgroups, designated low, moderate, and high DMI (LDMI, MDMI, and HDMI), according to actual DMI per day: ≤ 26 kg (n=27); >26 through 28.2 kg (n=28); and >28.2 kg (n=27). The phenotypic Pearson correlations among variables were analyzed, and the GLM procedure was used to test differences between subgroups. The relationships between milk and fat-corrected milk yields and the corresponding gross efficiencies were positive, whereas BW and gross production efficiency were negatively correlated. The marginal production efficiency from DM and energy consumed decreased with increasing DMI. The difference between BW gain as predicted by the National Research Council model (2001) and the present measurements increased with increasing DMI (r=0.68). The average calculated energy balances were 1.38, 2.28, and 4.20 Mcal/d (standard error of the mean=0.64) in the LDMI, MDMI, and HDMI groups, respectively. The marginal efficiency for milk yields from DMI or energy consumed was highest in LDMI, intermediate in MDMI, and lowest in HDMI. The predicted BW gains for the whole study period were 22.9, 37.9, and 75.8 kg for the LDMI, MDMI, and HDMI groups, respectively. The

  9. Future consequences of decreasing marginal production efficiency in the high-yielding dairy cow.

    PubMed

    Moallem, U

    2016-04-01

    The objectives were to examine the gross and marginal production efficiencies in high-yielding dairy cows and the future consequences on dairy industry profitability. Data from 2 experiments were used in across-treatments analysis (n=82 mid-lactation multiparous Israeli-Holstein dairy cows). Milk yields, body weights (BW), and dry matter intakes (DMI) were recorded daily. In both experiments, cows were fed a diet containing 16.5 to 16.6% crude protein and net energy for lactation (NEL) at 1.61 Mcal/kg of dry matter (DM). The means of milk yield, BW, DMI, NEL intake, and energy required for maintenance were calculated individually over the whole study, and used to calculate gross and marginal efficiencies. Data were analyzed in 2 ways: (1) simple correlation between variables; and (2) cows were divided into 3 subgroups, designated low, moderate, and high DMI (LDMI, MDMI, and HDMI), according to actual DMI per day: ≤ 26 kg (n=27); >26 through 28.2 kg (n=28); and >28.2 kg (n=27). The phenotypic Pearson correlations among variables were analyzed, and the GLM procedure was used to test differences between subgroups. The relationships between milk and fat-corrected milk yields and the corresponding gross efficiencies were positive, whereas BW and gross production efficiency were negatively correlated. The marginal production efficiency from DM and energy consumed decreased with increasing DMI. The difference between BW gain as predicted by the National Research Council model (2001) and the present measurements increased with increasing DMI (r=0.68). The average calculated energy balances were 1.38, 2.28, and 4.20 Mcal/d (standard error of the mean=0.64) in the LDMI, MDMI, and HDMI groups, respectively. The marginal efficiency for milk yields from DMI or energy consumed was highest in LDMI, intermediate in MDMI, and lowest in HDMI. The predicted BW gains for the whole study period were 22.9, 37.9, and 75.8 kg for the LDMI, MDMI, and HDMI groups, respectively. The

  10. High throughput and high yield nanofabrication of precisely designed gold nanohole arrays for fluorescence enhanced detection of biomarkers.

    PubMed

    Wong, Ten It; Han, Shan; Wu, Lin; Wang, Yi; Deng, Jie; Tan, Christina Yuan Ling; Bai, Ping; Loke, Yee Chong; Yang, Xin Da; Tse, Man Siu; Ng, Sum Huan; Zhou, Xiaodong

    2013-06-21

    Fluorescence excitation enhancement by plasmonic nanostructures such as gold nanohole arrays has been a hot topic in biosensing and bioimaging in recent years. However, the high throughput and high yield fabrication of precisely designed metal nanostructures for optimized fluorescence excitation remains a challenge. Our work is the first report combining nanopattern nickel mould fabrication and UV imprinting for gold nanostructure mass fabrication in high yield. We report our successful gold nanohole array mass fabrication on a 4'' glass wafer, by first fabricating a high fidelity nickel mould, then using the mould for UV nanoimprinting on a polymer coated on the glass, evaporating the gold film on the glass wafer, and lifting off the polymer to obtain a gold nanohole array on the glass. Our optimized process for wafer fabrication can achieve almost 100% yield from nanoimprinting to gold lift-off, while the fabricated nickel mould has >70% defect-free area with the rest having a few scattered defects. In our work, the size and pitch of the gold nanohole array are designed to enhance the fluorescent dye Alexa 647. When the fabricated gold nanohole array is used for prostate specific antigen (PSA) detection by establishing a sandwiched fluorescence assay on the gold surface, a detection limit of 100 pg ml(-1) is achieved, while with a same thickness of gold film, only 1 ng ml(-1) is detected. PMID:23645079

  11. High-yield oil palm expansion spares land at the expense of forests in the Peruvian Amazon

    NASA Astrophysics Data System (ADS)

    Gutiérrez-Vélez, Víctor H.; DeFries, Ruth; Pinedo-Vásquez, Miguel; Uriarte, María; Padoch, Christine; Baethgen, Walter; Fernandes, Katia; Lim, Yili

    2011-01-01

    High-yield agriculture potentially reduces pressure on forests by requiring less land to increase production. Using satellite and field data, we assessed the area deforested by industrial-scale high-yield oil palm expansion in the Peruvian Amazon from 2000 to 2010, finding that 72% of new plantations expanded into forested areas. In a focus area in the Ucayali region, we assessed deforestation for high- and smallholder low-yield oil palm plantations. Low-yield plantations accounted for most expansion overall (80%), but only 30% of their expansion involved forest conversion, contrasting with 75% for high-yield expansion. High-yield expansion minimized the total area required to achieve production but counter-intuitively at higher expense to forests than low-yield plantations. The results show that high-yield agriculture is an important but insufficient strategy to reduce pressure on forests. We suggest that high-yield agriculture can be effective in sparing forests only if coupled with incentives for agricultural expansion into already cleared lands.

  12. Wafer scale fabrication of carbon nanotube thin film transistors with high yield

    NASA Astrophysics Data System (ADS)

    Tian, Boyuan; Liang, Xuelei; Yan, Qiuping; Zhang, Han; Xia, Jiye; Dong, Guodong; Peng, Lianmao; Xie, Sishen

    2016-07-01

    Carbon nanotube thin film transistors (CNT-TFTs) are promising candidates for future high performance and low cost macro-electronics. However, most of the reported CNT-TFTs are fabricated in small quantities on a relatively small size substrate. The yield of large scale fabrication and the performance uniformity of devices on large size substrates should be improved before the CNT-TFTs reach real products. In this paper, 25 200 devices, with various geometries (channel width and channel length), were fabricated on 4-in. size ridged and flexible substrates. Almost 100% device yield were obtained on a rigid substrate with high out-put current (>8 μA/μm), high on/off current ratio (>105), and high mobility (>30 cm2/V.s). More importantly, uniform performance in 4-in. area was achieved, and the fabrication process can be scaled up. The results give us more confidence for the real application of the CNT-TFT technology in the near future.

  13. Most mutant OccR proteins that are defective in positive control hold operator DNA in a locked high-angle bend.

    PubMed

    Tsai, Ching-Sung; Chen, Chia-Sui; Winans, Stephen C

    2011-10-01

    OccR is a LysR-type transcriptional regulator of Agrobacterium tumefaciens that positively regulates the octopine catabolism operon of the Ti plasmid. Positive control of the occ genes occurs in response to octopine, a nutrient released from crown gall tumors. OccR also functions as an autorepressor in the presence or absence of octopine. OccR binds to a site between occQ and occR in the presence or absence of octopine, although octopine triggers a conformational change that shortens the DNA footprint and relaxes a DNA bend. In order to determine the roles of this conformational change in transcriptional activation, we isolated 11 OccR mutants that were defective in activation of the occQ promoter but were still capable of autorepression. The mutations in these mutants spanned most of the length of the protein. Two additional positive-control mutants were isolated using site-directed mutagenesis. Twelve mutant proteins displayed a high-angle DNA bend in the presence or absence of octopine. One mutant, the L26A mutant, showed ligand-responsive DNA binding similar to that of wild-type OccR and therefore must be impaired in a subsequent step in activation. PMID:21804007

  14. Characterization of neutron yield and x-ray spectra of a High Flux Neutron Generator (HFNG)

    NASA Astrophysics Data System (ADS)

    Nnamani, Nnaemeka; HFNG Collaboration

    2015-04-01

    The High Flux Neutron Generator (HFNG) is a DD plasma-based source, with a self-loading target intended for fundamental science and engineering applications, including 40 Ar/39 Ar geochronology, neutron cross section measurements, and radiation hardness testing of electronics. Our first estimate of the neutron yield, based on the population of the 4.486 hour 115 In isomer gave a neutron yield of the order 108 n/sec; optimization is ongoing to achieve the design target of 1011 n/sec. Preliminary x-ray spectra showed prominent energy peaks which are likely due to atomic line-emission from back-streaming electrons accelerated up to 100 keV impinging on various components of the HFNG chamber. Our x-ray and neutron diagnostics will aid us as we continue to evolve the design to suppress back-streaming electrons, necessary to achieve higher plasma beam currents, and thus higher neutron flux. This talk will focus on the characterization of the neutron yield and x-ray spectra during our tests. A collimation system is being installed near one of the chamber ports for improved observation of the x-ray spectra. This work is supported by NSF Grant No. EAR-0960138, U.S. DOE LBNL Contract No. DE-AC02-05CH11231, U.S. DOE LLNL Contract No. DE-AC52-07NA27344, and the UC Office of the President Award 12-LR-238745.

  15. Sortase A as a tool for high-yield histatin cyclization.

    PubMed

    Bolscher, Jan G M; Oudhoff, Menno J; Nazmi, Kamran; Antos, John M; Guimaraes, Carla P; Spooner, Eric; Haney, Evan F; Garcia Vallejo, Juan J; Vogel, Hans J; van't Hof, Wim; Ploegh, Hidde L; Veerman, Enno C I

    2011-08-01

    Cyclic peptides are highly valued tools in biomedical research. In many cases, they show higher receptor affinity, enhanced biological activity, and improved serum stability. Technical difficulties in producing cyclic peptides, especially larger ones, in appreciable yields have precluded a prolific use in biomedical research. Here, we describe a novel and efficient cyclization method that uses the peptidyl-transferase activity of the Staphylococcus aureus enzyme sortase A to cyclize linear synthetic precursor peptides. As a model, we used histatin 1, a 38-mer salivary peptide with motogenic activity. Chemical cyclization of histatin 1 resulted in ≤ 3% yields, whereas sortase-mediated cyclization provided a yield of >90%. The sortase-cyclized peptide displayed a maximum wound closure activity at 10 nM, whereas the linear peptide displayed maximal activity at 10 μM. Circular dichroism and NMR spectroscopic analysis of the linear and cyclic peptide in solution showed no evidence for conformational changes, suggesting that structural differences due to cyclization only became manifest when these peptides were located in the binding domain of the receptor. The sortase-based cyclization technology provides a general method for easy and efficient manufacturing of large cyclic peptides.

  16. Cacao Cultivation under Diverse Shade Tree Cover Allows High Carbon Storage and Sequestration without Yield Losses

    PubMed Central

    Abou Rajab, Yasmin; Leuschner, Christoph; Barus, Henry; Tjoa, Aiyen; Hertel, Dietrich

    2016-01-01

    One of the main drivers of tropical forest loss is their conversion to oil palm, soy or cacao plantations with low biodiversity and greatly reduced carbon storage. Southeast Asian cacao plantations are often established under shade tree cover, but are later converted to non-shaded monocultures to avoid resource competition. We compared three co-occurring cacao cultivation systems (3 replicate stands each) with different shade intensity (non-shaded monoculture, cacao with the legume Gliricidia sepium shade trees, and cacao with several shade tree species) in Sulawesi (Indonesia) with respect to above- and belowground biomass and productivity, and cacao bean yield. Total biomass C stocks (above- and belowground) increased fivefold from the monoculture to the multi-shade tree system (from 11 to 57 Mg ha-1), total net primary production rose twofold (from 9 to 18 Mg C ha-1 yr-1). This increase was associated with a 6fold increase in aboveground biomass, but only a 3.5fold increase in root biomass, indicating a clear shift in C allocation to aboveground tree organs with increasing shade for both cacao and shade trees. Despite a canopy cover increase from 50 to 93%, cacao bean yield remained invariant across the systems (variation: 1.1–1.2 Mg C ha-1 yr-1). The monocultures had a twice as rapid leaf turnover suggesting that shading reduces the exposure of cacao to atmospheric drought, probably resulting in greater leaf longevity. Thus, contrary to general belief, cacao bean yield does not necessarily decrease under shading which seems to reduce physical stress. If planned properly, cacao plantations under a shade tree cover allow combining high yield with benefits for carbon sequestration and storage, production system stability under stress, and higher levels of animal and plant diversity. PMID:26927428

  17. Cacao Cultivation under Diverse Shade Tree Cover Allows High Carbon Storage and Sequestration without Yield Losses.

    PubMed

    Abou Rajab, Yasmin; Leuschner, Christoph; Barus, Henry; Tjoa, Aiyen; Hertel, Dietrich

    2016-01-01

    One of the main drivers of tropical forest loss is their conversion to oil palm, soy or cacao plantations with low biodiversity and greatly reduced carbon storage. Southeast Asian cacao plantations are often established under shade tree cover, but are later converted to non-shaded monocultures to avoid resource competition. We compared three co-occurring cacao cultivation systems (3 replicate stands each) with different shade intensity (non-shaded monoculture, cacao with the legume Gliricidia sepium shade trees, and cacao with several shade tree species) in Sulawesi (Indonesia) with respect to above- and belowground biomass and productivity, and cacao bean yield. Total biomass C stocks (above- and belowground) increased fivefold from the monoculture to the multi-shade tree system (from 11 to 57 Mg ha-1), total net primary production rose twofold (from 9 to 18 Mg C ha-1 yr-1). This increase was associated with a 6fold increase in aboveground biomass, but only a 3.5fold increase in root biomass, indicating a clear shift in C allocation to aboveground tree organs with increasing shade for both cacao and shade trees. Despite a canopy cover increase from 50 to 93%, cacao bean yield remained invariant across the systems (variation: 1.1-1.2 Mg C ha-1 yr-1). The monocultures had a twice as rapid leaf turnover suggesting that shading reduces the exposure of cacao to atmospheric drought, probably resulting in greater leaf longevity. Thus, contrary to general belief, cacao bean yield does not necessarily decrease under shading which seems to reduce physical stress. If planned properly, cacao plantations under a shade tree cover allow combining high yield with benefits for carbon sequestration and storage, production system stability under stress, and higher levels of animal and plant diversity.

  18. Cacao Cultivation under Diverse Shade Tree Cover Allows High Carbon Storage and Sequestration without Yield Losses.

    PubMed

    Abou Rajab, Yasmin; Leuschner, Christoph; Barus, Henry; Tjoa, Aiyen; Hertel, Dietrich

    2016-01-01

    One of the main drivers of tropical forest loss is their conversion to oil palm, soy or cacao plantations with low biodiversity and greatly reduced carbon storage. Southeast Asian cacao plantations are often established under shade tree cover, but are later converted to non-shaded monocultures to avoid resource competition. We compared three co-occurring cacao cultivation systems (3 replicate stands each) with different shade intensity (non-shaded monoculture, cacao with the legume Gliricidia sepium shade trees, and cacao with several shade tree species) in Sulawesi (Indonesia) with respect to above- and belowground biomass and productivity, and cacao bean yield. Total biomass C stocks (above- and belowground) increased fivefold from the monoculture to the multi-shade tree system (from 11 to 57 Mg ha-1), total net primary production rose twofold (from 9 to 18 Mg C ha-1 yr-1). This increase was associated with a 6fold increase in aboveground biomass, but only a 3.5fold increase in root biomass, indicating a clear shift in C allocation to aboveground tree organs with increasing shade for both cacao and shade trees. Despite a canopy cover increase from 50 to 93%, cacao bean yield remained invariant across the systems (variation: 1.1-1.2 Mg C ha-1 yr-1). The monocultures had a twice as rapid leaf turnover suggesting that shading reduces the exposure of cacao to atmospheric drought, probably resulting in greater leaf longevity. Thus, contrary to general belief, cacao bean yield does not necessarily decrease under shading which seems to reduce physical stress. If planned properly, cacao plantations under a shade tree cover allow combining high yield with benefits for carbon sequestration and storage, production system stability under stress, and higher levels of animal and plant diversity. PMID:26927428

  19. Unexpected high yields of carbonyl and peroxide products of aqueous isoprene ozonolysis and implications

    NASA Astrophysics Data System (ADS)

    Wang, H. L.; Huang, D.; Zhang, X.; Zhao, Y.; Chen, Z. M.

    2012-03-01

    The aqueous phase reaction of volatile organic compounds (VOCs) has not been considered in most analyses of atmospheric chemical processes. However, some experimental evidence has shown that, compared to the corresponding gas phase reaction, the aqueous chemical processes of VOCs in the bulk solutions and surfaces of ambient wet particles (cloud, fog, and wet aerosols) may potentially contribute to the products and formation of secondary organic aerosol (SOA). In the present study, we performed a laboratory experiment of the aqueous ozonolysis of isoprene at different pHs (3-7) and temperatures (4-25 °C). We detected three important kinds of products, including carbonyl compounds, peroxide compounds, and organic acids. Our results showed that the molar yields of these products were nearly independent of the investigated pHs and temperatures. These products included (1) carbonyls: 56.7 ± 6.7% formaldehyde, 42.8 ± 2.5% methacrolein (MAC), and 57.7 ± 3.4% methyl vinyl ketone (MVK); (2) peroxides: 53.4 ± 4.1% hydrogen peroxide (H2O2) and 15.1 ± 3.1% hydroxylmethyl hydroperoxide (HMHP); and (3) organic acids: undetectable (< 1% estimated by the detection limit). Based on the amounts of products formed and the isoprene consumed, the total carbon yield was estimated to be 95 ± 4%. This implied that most of the products in the reaction system were detected. Of note, the combined yields of both MAC + MVK and H2O2 + HMHP in the aqueous isoprene ozonolysis were much higher than those observed in the corresponding gas phase reaction. We suggested that these unexpected high yields of carbonyls and peroxides were related to the greater capability of condensed water, compared to water vapor, to stabilize energy-rich Criegee radicals. This aqueous ozonolysis of isoprene (and possibly other biogenic VOCs) could potentially occur on the surfaces of ambient wet particles and plants. Moreover, the high-yield carbonyl and peroxide products might provide a considerable source of

  20. High night temperatures during grain number determination reduce wheat and barley grain yield: a field study.

    PubMed

    García, Guillermo A; Dreccer, M Fernanda; Miralles, Daniel J; Serrago, Román A

    2015-11-01

    Warm nights are a widespread predicted feature of climate change. This study investigated the impact of high night temperatures during the critical period for grain yield determination in wheat and barley crops under field conditions, assessing the effects on development, growth and partitioning crop-level processes driving grain number per unit area (GN). Experiments combined: (i) two contrasting radiation and temperature environments: late sowing in 2011 and early sowing in 2013, (ii) two well-adapted crops with similar phenology: bread wheat and two-row malting barley and (iii) two temperature regimes: ambient and high night temperatures. The night temperature increase (ca. 3.9 °C in both crops and growing seasons) was achieved using purpose-built heating chambers placed on the crop at 19:000 hours and removed at 7:00 hours every day from the third detectable stem node to 10 days post-flowering. Across growing seasons and crops, the average minimum temperature during the critical period ranged from 11.2 to 17.2 °C. Wheat and barley grain yield were similarly reduced under warm nights (ca. 7% °C(-1) ), due to GN reductions (ca. 6% °C(-1) ) linked to a lower number of spikes per m(2) . An accelerated development under high night temperatures led to a shorter critical period duration, reducing solar radiation capture with negative consequences for biomass production, GN and therefore, grain yield. The information generated could be used as a starting point to design management and/or breeding strategies to improve crop adaptation facing climate change.

  1. High night temperatures during grain number determination reduce wheat and barley grain yield: a field study.

    PubMed

    García, Guillermo A; Dreccer, M Fernanda; Miralles, Daniel J; Serrago, Román A

    2015-11-01

    Warm nights are a widespread predicted feature of climate change. This study investigated the impact of high night temperatures during the critical period for grain yield determination in wheat and barley crops under field conditions, assessing the effects on development, growth and partitioning crop-level processes driving grain number per unit area (GN). Experiments combined: (i) two contrasting radiation and temperature environments: late sowing in 2011 and early sowing in 2013, (ii) two well-adapted crops with similar phenology: bread wheat and two-row malting barley and (iii) two temperature regimes: ambient and high night temperatures. The night temperature increase (ca. 3.9 °C in both crops and growing seasons) was achieved using purpose-built heating chambers placed on the crop at 19:000 hours and removed at 7:00 hours every day from the third detectable stem node to 10 days post-flowering. Across growing seasons and crops, the average minimum temperature during the critical period ranged from 11.2 to 17.2 °C. Wheat and barley grain yield were similarly reduced under warm nights (ca. 7% °C(-1) ), due to GN reductions (ca. 6% °C(-1) ) linked to a lower number of spikes per m(2) . An accelerated development under high night temperatures led to a shorter critical period duration, reducing solar radiation capture with negative consequences for biomass production, GN and therefore, grain yield. The information generated could be used as a starting point to design management and/or breeding strategies to improve crop adaptation facing climate change. PMID:26111197

  2. Delta-Doping at Wafer Level for High Throughput, High Yield Fabrication of Silicon Imaging Arrays

    NASA Technical Reports Server (NTRS)

    Hoenk, Michael E. (Inventor); Nikzad, Shoulch (Inventor); Jones, Todd J. (Inventor); Greer, Frank (Inventor); Carver, Alexander G. (Inventor)

    2014-01-01

    Systems and methods for producing high quantum efficiency silicon devices. A silicon MBE has a preparation chamber that provides for cleaning silicon surfaces using an oxygen plasma to remove impurities and a gaseous (dry) NH3 + NF3 room temperature oxide removal process that leaves the silicon surface hydrogen terminated. Silicon wafers up to 8 inches in diameter have devices that can be fabricated using the cleaning procedures and MBE processing, including delta doping.

  3. A high yield technique for freeze-fracturing of small fractions of isolated cells.

    PubMed

    Falcieri, E; Mariani, A R; Del Coco, R; Facchini, A; Maraldi, N M

    1988-07-01

    A simple, high-yield technique for the freeze-fracturing of small amounts of isolated cells is described. A drop of cells fixed in suspension is deposited on a polylysine-treated coverslip, forming a monolayer through electrostatic forces. After cryoprotection, the coverslip is inverted on a gold carrier covered with Vinol and then frozen in liquid nitrogen. The monolayer will be fractured by advancing the knife under the coverslip. Large areas of cell surface can be exposed despite their low number, such as that obtainable after cell sorting by flow cytometry. PMID:3179998

  4. A High Yield Synthesis of Chalcopyrite CuInS2 Nanoparticles with Exceptional Size Control

    SciTech Connect

    Sun, Chivin; Gardner, Joseph S.; Shurdha, Endrit; Margulieux, Kelsey R.; Westover, Richard D.; Lau, Lisa D.; Long, Gary; Bajracharya, Cyril; Wang, Chong M.; Thurber, Aaron P.; Punnoose, Alex; Rodriguez, Rene G.; Pak, Joshua J.

    2009-12-29

    Various I-III-VI2 semiconductor materials have been identified as promising photovoltaic materials [1, 2]. Recently, quantum dot (QD) based solar cells have attracted much attention due to their potential to replace thin film devices [3-5]. One of the major advantages of employing QDs is by simply changing the particle size they can be tuned to absorb specific wavelengths ranging from visible to infrared wavelengths [6]. Furthermore, with careful design of photovoltaic (PV) devices incorporating various sizes of nanoparticles in multiple layers, one may achieve increased solar energy absorption in one device [7, 8]. In order to facilitate QD based multilayer devices, synthetic strategies that can deliver QDs in high yields with precise size control are essential. One of the strategies to prepare QDs is to prepare nanoparticles from molecular single source precursors (SSPs), which contain all necessary elements in a single molecule. In recent years, there have been several reports on the formation of CuInS2 nanoparticles through the decomposition of SSPs using thermolysis [9-14], photolysis [18], and microwave irradiation [16]. However, many of these procedures require a combination of long reaction times (10 to 24 hours) and high reaction temperatures (often exceeding 200 °C) with very little information regarding overall yields. Microwave-assisted growth of nanoparticles is generally favorable over traditional thermolysis as microwave irradiation overcomes local intermediaries and increases the microscopic temperature of the reaction [17] thus exhibiting greater homogeneity in the overall reaction temperature. This allows for nanoparticles with diameters of a few nanometers can be prepared [18], dramatic decreases in reaction times, improved product purities, all forms of precursors can be used, and reactions exhibit high reproducibility and yields [19]. For CuInS2 QDs, the Wannier-Mott bulk exciton radius is approximately 8 nm with a bandgap of 1.45 eV and QDs

  5. Very high CO2 reduces photosynthesis, dark respiration and yield in wheat.

    PubMed

    Reuveni, J; Bugbee, B

    1997-10-01

    Although terrestrial CO2 concentrations, [CO2] are not expected to reach 1000 micromoles mol-1 for many decades, CO2 levels in closed systems such as growth chambers and glasshouses, can easily exceed this concentration. CO2 levels in life support systems in space can exceed 10000 micromoles mol-1 (1%). Here we studied the effect of six CO2 concentrations, from ambient up to 10000 micromoles mol-1, on seed yield, growth and gas exchange of two wheat cultivars (USU-Apogee and Veery-l0). Elevating [CO2] from 350 to 1000 micromoles mol-1 increased seed yield (by 33%), vegetative biomass (by 25%) and number of heads m-2 (by 34%) of wheat plants. Elevation of [CO2] from 1000 to 10000 micromoles mol-1 decreased seed yield (by 37%), harvest index (by 14%), mass per seed (by 9%) and number of seeds per head (by 29%). This very high [CO2] had a negligible, non-significant effect on vegetative biomass, number of heads m-2 and seed mass per head. A sharp decrease in seed yield, harvest index and seeds per head occurred by elevating [CO2] from 1000 to 2600 micromoles mol-1. Further elevation of [CO2] from 2600 to 10000 micromoles mol-1 caused a further but smaller decrease. The effect of CO2 on both wheat cultivars was similar for all growth parameters. Similarly there were no differences in the response to high [CO2] between wheat grown hydroponically in growth chambers under fluorescent lights and those grown in soilless media in a glasshouse under sunlight and high pressure sodium lamps. There was no correlation between high [CO2] and ethylene production by flag leaves or by wheat heads. Therefore, the reduction in seed set in wheat plants is not mediated by ethylene. The photosynthetic rate of whole wheat plants was 8% lower and dark respiration of the wheat heads 25% lower when exposed to 2600 micromoles mol-1 CO2 compared to ambient [CO2]. It is concluded that the reduction in the seed set can be mainly explained by the reduction in the dark respiration in wheat heads

  6. Carbon dots with high fluorescence quantum yield: the fluorescence originates from organic fluorophores.

    PubMed

    Shi, Lei; Yang, Jian Hai; Zeng, Hai Bo; Chen, Yong Mei; Yang, Sheng Chun; Wu, Chao; Zeng, Hao; Yoshihito, Osada; Zhang, Qiqing

    2016-08-14

    In this contribution, we have shown that the organic fluorophores, 5-oxo-3,5-dihydro-2H-thiazolo [3,2-a] pyridine-3,7-dicarboxylic acid (TPDCA) and 5-oxo-3,5-dihydro-2H-thiazolo [3,2-a] pyridine-7-carboxylic acid (TPCA), are the main ingredients and fluorescence origins of N,S-CDs via systematic analyses. It inspires us to deeply analyze and understand the fluorescence origins of carbon dots with high fluorescence quantum yields, which will expand their applications. PMID:27426926

  7. Very high CO2 reduces photosynthesis, dark respiration and yield in wheat

    NASA Technical Reports Server (NTRS)

    Reuveni, J.; Bugbee, B.

    1997-01-01

    Although terrestrial CO2 concentrations, [CO2] are not expected to reach 1000 micromoles mol-1 for many decades, CO2 levels in closed systems such as growth chambers and glasshouses, can easily exceed this concentration. CO2 levels in life support systems in space can exceed 10000 micromoles mol-1 (1%). Here we studied the effect of six CO2 concentrations, from ambient up to 10000 micromoles mol-1, on seed yield, growth and gas exchange of two wheat cultivars (USU-Apogee and Veery-l0). Elevating [CO2] from 350 to 1000 micromoles mol-1 increased seed yield (by 33%), vegetative biomass (by 25%) and number of heads m-2 (by 34%) of wheat plants. Elevation of [CO2] from 1000 to 10000 micromoles mol-1 decreased seed yield (by 37%), harvest index (by 14%), mass per seed (by 9%) and number of seeds per head (by 29%). This very high [CO2] had a negligible, non-significant effect on vegetative biomass, number of heads m-2 and seed mass per head. A sharp decrease in seed yield, harvest index and seeds per head occurred by elevating [CO2] from 1000 to 2600 micromoles mol-1. Further elevation of [CO2] from 2600 to 10000 micromoles mol-1 caused a further but smaller decrease. The effect of CO2 on both wheat cultivars was similar for all growth parameters. Similarly there were no differences in the response to high [CO2] between wheat grown hydroponically in growth chambers under fluorescent lights and those grown in soilless media in a glasshouse under sunlight and high pressure sodium lamps. There was no correlation between high [CO2] and ethylene production by flag leaves or by wheat heads. Therefore, the reduction in seed set in wheat plants is not mediated by ethylene. The photosynthetic rate of whole wheat plants was 8% lower and dark respiration of the wheat heads 25% lower when exposed to 2600 micromoles mol-1 CO2 compared to ambient [CO2]. It is concluded that the reduction in the seed set can be mainly explained by the reduction in the dark respiration in wheat heads

  8. Neutron temporal diagnostic for high-yield deuterium-tritium cryogenic implosions on OMEGA

    NASA Astrophysics Data System (ADS)

    Stoeckl, C.; Boni, R.; Ehrne, F.; Forrest, C. J.; Glebov, V. Yu.; Katz, J.; Lonobile, D. J.; Magoon, J.; Regan, S. P.; Shoup, M. J.; Sorce, A.; Sorce, C.; Sangster, T. C.; Weiner, D.

    2016-05-01

    A next-generation neutron temporal diagnostic (NTD) capable of recording high-quality data for the highest anticipated yield cryogenic deuterium-tritium (DT) implosion experiments was recently installed at the Omega Laser Facility. A high-quality measurement of the neutron production width is required to determine the hot-spot pressure achieved in inertial confinement fusion experiments—a key metric in assessing the quality of these implosions. The design of this NTD is based on a fast-rise-time plastic scintillator, which converts the neutron kinetic energy to 350- to 450-nm-wavelength light. The light from the scintillator inside the nose-cone assembly is relayed ˜16 m to a streak camera in a well-shielded location. An ˜200× reduction in neutron background was observed during the first high-yield DT cryogenic implosions compared to the current NTD installation on OMEGA. An impulse response of ˜40 ± 10 ps was measured in a dedicated experiment using hard x-rays from a planar target irradiated with a 10-ps short pulse from the OMEGA EP laser. The measured instrument response includes contributions from the scintillator rise time, optical relay, and streak camera.

  9. A Permanent-Magnet Microwave Ion Source for a Compact High-Yield Neutron Generator

    SciTech Connect

    Waldmann, Ole; Ludewigt, Bernhard

    2010-10-11

    We present recent work on the development of a microwave ion source that will be used in a high-yield compact neutron generator for active interrogation applications. The sealed tube generator will be capable of producing high neutron yields, 5x1011 n/s for D-T and ~;;1x1010 n/s for D-D reactions, while remaining transportable. We constructed a microwave ion source (2.45 GHz) with permanent magnets to provide the magnetic field strength of 87.5 mT necessary for satisfying the electron cyclotron resonance (ECR) condition. Microwave ion sources can produce high extracted beam currents at the low gas pressures required for sealed tube operation and at lower power levels than previously used RF-driven ion sources. A 100 mA deuterium/tritium beam will be extracted through a large slit (60x6 mm2) to spread the beam power over a larger target area. This paper describes the design of the permanent-magnet microwave ion source and discusses the impact of the magnetic field design on the source performance. The required equivalent proton beam current density of 40 mA/cm2 was extracted at a moderate microwave power of 400 W with an optimized magnetic field.

  10. RNA isolation from loquat and other recalcitrant woody plants with high quality and yield.

    PubMed

    Morante-Carriel, Jaime; Sellés-Marchart, Susana; Martínez-Márquez, Ascensión; Martínez-Esteso, María José; Luque, Ignacio; Bru-Martínez, Roque

    2014-05-01

    RNA isolation is difficult in plants that contain large amounts of polysaccharides and polyphenol compounds. To date, no commercial kit has been developed for the isolation of high-quality RNA from tissues with these characteristics, especially for fruit. The common protocols for RNA isolation are tedious and usually result in poor yields when applied to recalcitrant plant tissues. Here an efficient RNA isolation protocol based on cetyltrimethylammonium bromide (CTAB) and two successive precipitations with 10 M lithium chloride (LiCl) was developed specifically for loquat fruits, but it was proved to work efficiently in other tissues of loquat and woody plants. The RNA isolated by this improved protocol was not only of high purity and integrity (A260/A280 ratios ranged from 1.90 to 2.04 and A260/A230 ratios were>2.0) but also of high yield (up to 720 μg on average [coefficient of variation=21%] total RNA per gram fresh tissue). The protocol was tested on loquat fruit (different stages of development, postharvest, ripening, and bruising), leaf, root, flower, stem, and bud; quince fruit and root; grapevine cells in liquid culture; and rose petals. The RNA obtained with this method is amenable to enzymatic treatments and can be efficiently applied for research on gene characterization, expression, and function.

  11. Propagation method for persistent high yield of diverse Listeria phages on permissive hosts at refrigeration temperatures.

    PubMed

    Radford, Devon R; Ahmadi, Hanie; Leon-Velarde, Carlos G; Balamurugan, Sampathkumar

    2016-10-01

    The efficient production of a high concentration of bacteriophage in large volumes has been a limiting factor in the exploration of the true potential of these organisms for biotechnology, agriculture and medicine. Traditional methods focus on generating small volumes of highly concentrated samples as the end product of extensive mechanical and osmotic processing. To function at an industrial scale mandates extensive investment in infrastructure and input materials not feasible for many smaller facilities. To address this, we developed a novel, scalable, generic method for producing significantly higher titer psychrophilic phage (P < 2.0 × 10(-6)), 2- to 4-fold faster than traditional methods. We generate renewable high yields from single source cultures by propagating phage under refrigeration conditions in which Listeria, Yersinia and their phages grow in equilibrium. Diverse Yersinia and Listeria phages tested yielded averages of 3.49 × 10(8) to 3.36 × 10(12) PFU/ml/day compared to averages of 1.28 × 10(5) to 1.30 × 10(10) PFU/ml/day by traditional methods. Host growth and death kinetics made this method ineffective for extended propagation of mesophilic phages.

  12. Propagation method for persistent high yield of diverse Listeria phages on permissive hosts at refrigeration temperatures.

    PubMed

    Radford, Devon R; Ahmadi, Hanie; Leon-Velarde, Carlos G; Balamurugan, Sampathkumar

    2016-10-01

    The efficient production of a high concentration of bacteriophage in large volumes has been a limiting factor in the exploration of the true potential of these organisms for biotechnology, agriculture and medicine. Traditional methods focus on generating small volumes of highly concentrated samples as the end product of extensive mechanical and osmotic processing. To function at an industrial scale mandates extensive investment in infrastructure and input materials not feasible for many smaller facilities. To address this, we developed a novel, scalable, generic method for producing significantly higher titer psychrophilic phage (P < 2.0 × 10(-6)), 2- to 4-fold faster than traditional methods. We generate renewable high yields from single source cultures by propagating phage under refrigeration conditions in which Listeria, Yersinia and their phages grow in equilibrium. Diverse Yersinia and Listeria phages tested yielded averages of 3.49 × 10(8) to 3.36 × 10(12) PFU/ml/day compared to averages of 1.28 × 10(5) to 1.30 × 10(10) PFU/ml/day by traditional methods. Host growth and death kinetics made this method ineffective for extended propagation of mesophilic phages. PMID:27287043

  13. Neutron temporal diagnostic for high-yield deuterium-tritium cryogenic implosions on OMEGA

    DOE PAGESBeta

    Stoeckl, C.; Boni, R.; Ehrne, F.; Forrest, C. J.; Glebov, V. Yu.; Katz, J.; Lonobile, D. J.; Magoon, J.; Regan, S. P.; Shoup, III, M. J.; et al

    2016-05-10

    A next-generation neutron temporal diagnostic (NTD) capable of recording high-quality data for the highest anticipated yield cryogenic DT implosion experiments was recently installed at the Omega Laser Facility. A high-quality measurement of the neutron production width is required to determine the hot-spot pressure achieved in inertial confinement fusion experiments—a key metric in assessing the quality of these implosions. The design of this NTD is based on a fast-rise-time plastic scintillator, which converts the neutron kinetic energy to 350- to 450-nm-wavelength light. The light from the scintillator inside the nose-cone assembly is relayed ~16 m to a streak camera in amore » well-shielded location. An ~200× reduction in neutron background was observed during the first high-yield DT cryogenic implosions compared to the current NTD installation on OMEGA. An impulse response of ~40±10 ps was measured in a dedicated experiment using hard x rays from a planar target irradiated with a 10-ps short pulse from the OMEGA EP laser. Furthermore, the measured instrument response includes contributions from the scintillator rise time, optical relay, and streak camera.« less

  14. High yields of 2,3-butanediol and mannitol in Lactococcus lactis through engineering of NAD⁺ cofactor recycling.

    PubMed

    Gaspar, Paula; Neves, Ana Rute; Gasson, Michael J; Shearman, Claire A; Santos, Helena

    2011-10-01

    Manipulation of NADH-dependent steps, and particularly disruption of the las-located lactate dehydrogenase (ldh) gene in Lactococcus lactis, is common to engineering strategies envisaging the accumulation of reduced end products other than lactate. Reverse transcription-PCR experiments revealed that three out of the four genes assigned to lactate dehydrogenase in the genome of L. lactis, i.e., the ldh, ldhB, and ldhX genes, were expressed in the parental strain MG1363. Given that genetic redundancy is often a major cause of metabolic instability in engineered strains, we set out to develop a genetically stable lactococcal host tuned for the production of reduced compounds. Therefore, the ldhB and ldhX genes were sequentially deleted in L. lactis FI10089, a strain with a deletion of the ldh gene. The single, double, and triple mutants, FI10089, FI10089ΔldhB, and FI10089ΔldhBΔldhX, showed similar growth profiles and displayed mixed-acid fermentation, ethanol being the main reduced end product. Hence, the alcohol dehydrogenase-encoding gene, the adhE gene, was inactivated in FI10089, but the resulting strain reverted to homolactic fermentation due to induction of the ldhB gene. The three lactate dehydrogenase-deficient mutants were selected as a background for the production of mannitol and 2,3-butanediol. Pathways for the biosynthesis of these compounds were overexpressed under the control of a nisin promoter, and the constructs were analyzed with respect to growth parameters and product yields under anaerobiosis. Glucose was efficiently channeled to mannitol (maximal yield, 42%) or to 2,3-butanediol (maximal yield, 67%). The theoretical yield for 2,3-butanediol was achieved. We show that FI10089ΔldhB is a valuable basis for engineering strategies aiming at the production of reduced compounds.

  15. Effective catalytic conversion of cellulose into high yields of methyl glucosides over sulfonated carbon based catalyst.

    PubMed

    Dora, Sambha; Bhaskar, Thallada; Singh, Rawel; Naik, Desavath Viswanatha; Adhikari, Dilip Kumar

    2012-09-01

    An amorphous carbon based catalyst was prepared by sulfonation of the bio-char obtained from fast pyrolysis (N(2) atm; ≈ 550°C) of biomass. The sulfonated carbon catalyst contained high acidity of 6.28 mmol/g as determined by temperature programmed desorption of ammonia of sulfonated carbon catalyst and exhibited high catalytic performance for the hydrolysis of cellulose. Amorphous carbon based catalyst containing -SO(3)H groups was successfully tested and the complete conversion of cellulose in methanol at moderate temperatures with high yields ca. ≥ 90% of α, β-methyl glucosides in short reaction times was achieved. The methyl glucosides formed in methanol are more stable for further conversion than the products formed in water. The carbon catalyst was demonstrated to be stable for five cycles with slight loss in catalytic activity. The utilization of bio-char as a sulfonated carbon catalyst provides a green and efficient process for cellulose conversion. PMID:22776237

  16. Improvement of bleached wheat straw pulp properties by using aspen high-yield pulp.

    PubMed

    Zhang, Hongjie; Li, Jianguo; Hu, Huiren; He, Zhibin; Ni, Yonghao

    2012-09-01

    The bleached wheat straw pulp (BWSP) accounts for about 25% of the virgin fiber supply in the Chinese Pulp and Paper Industry. As a non-wood chemical pulp, BWSP is known to have low bulk, low light scattering coefficient and poor drainage due to its high content of parenchyma cells. In this study, a high-quality aspen high-yield pulp (HYP) was used to improve the BWSP properties at the laboratory scale. The results indicate that adding 5-20% aspen HYP into unrefined or refined BWSP can minimize many of the drawbacks associated with the BWSP: improving its drainage, bulk, light scattering coefficient and opacity. The addition of a small amount (up to 20%) of aspen HYP can also significantly increase the tear index of BWSP with only a slight decrease of the tensile index.

  17. High-yield growth of carbon nanofilaments on nickel foam using nickel-tin intermetallic catalysts.

    PubMed

    Jeong, Namjo; Hwang, Kyo Sik; Yang, Seung Cheol

    2014-10-01

    The integration of nanomaterials into macroscopic structures is of importance to their practical use. We report the direct synthesis of carbon nanofilaments on Ni foam using Ni-Sn intermetallic nanoparticles. The use of SnO2 nanoparticles was highly effective for the high-yield growth of carbon nanofilaments without the occurrence of surface breakup, resulting from excessive carbon accumulation in the Ni foam. Carbon nanofilaments with a diameter of 50 nm were synthesized and contained Ni3Sn nanoparticles at the tip, indicating a tip-growth mechanism. Higher vacuum conditions led to the growth of highly crystalline carbon nanofilaments. The results obtained using different sources of hydrocarbon revealed that in contrast to C2H2, CH4 or C3H8 did not induce carbon nanofilament formation on Ni foam.

  18. High Yield Sample Preconcentration Using a Highly Ion-conductive Charge-selective Polymer

    PubMed Central

    Chun, Honggu; Chung, Taek Dong; Ramsey, J. Michael

    2011-01-01

    The development and analysis of a microfluidic sample preconcentration system using a highly ion-conductive charge-selective polymer (poly-AMPS) is reported. The preconcentration is based on the phenomenon of concentration polarization which develops at the boundaries of the poly-AMPS with buffer solutions. A negatively charged polymer, poly-AMPS, positioned between two microchannels efficiently extracts cations through its large cross section, resulting in efficient anion sample preconcentration. The present work includes the development of a robust polymer that is stable over a wide range of buffers with varying chemical compositions. The sample preconcentration effect remains linear to over 3 mM (0.15 pmol) and 500 μM (15 fmol) for fluorescein and TRITC-tagged albumin solutions, respectively. The system can potentially be used for concentrating proteins on microfluidic devices with subsequent analysis for proteomic applications. PMID:20575520

  19. High-resolution mapping and genetic characterization of the Lazy-2 gravitropic mutant of tomato

    NASA Technical Reports Server (NTRS)

    Behringer, F. J.; Lomax, T. L.

    1999-01-01

    Mutation of the Lazy-2 (Lz-2) gene in tomato (Lycopersicon esculentum mill.) produces a phytochrome-dependent reversal of shoot gravitropism, providing a unique genetic resource for investigating how signals from light modulate gravitropism. We mapped the Lz-2 gene using RFLPs and a PCR-based technique to assess the feasibility of positional cloning. Analysis of a 1338 plant backcross population between L. esculentum and L. pennellii placed Lz-2 within a 1.2 cM interval on chromosome 5, 0.4 cM from TG504-CT201A interval. The inabililty to resolve these markers indicates that Lz-2 resides in a centromeric region in which recombination is highly suppressed. Lazy-2 is tightly linked to but does not encode the gene for ACC4, an enzyme involved in ethylene biosynthesis. We also observed that Lz-2 is partially dominant under certain conditions and stages of development.

  20. High process yield rates of thermoplastic nanofluidic devices using a hybrid thermal assembly technique.

    PubMed

    Uba, Franklin I; Hu, Bo; Weerakoon-Ratnayake, Kumuditha; Oliver-Calixte, Nyote; Soper, Steven A

    2015-02-21

    Over the past decade, thermoplastics have been used as alternative substrates to glass and Si for microfluidic devices because of the diverse and robust fabrication protocols available for thermoplastics that can generate high production rates of the desired structures at low cost and with high replication fidelity, the extensive array of physiochemical properties they possess, and the simple surface activation strategies that can be employed to tune their surface chemistry appropriate for the intended application. While the advantages of polymer microfluidics are currently being realized, the evolution of thermoplastic-based nanofluidic devices is fraught with challenges. One challenge is assembly of the device, which consists of sealing a cover plate to the patterned fluidic substrate. Typically, channel collapse or substrate dissolution occurs during assembly making the device inoperable resulting in low process yield rates. In this work, we report a low temperature hybrid assembly approach for the generation of functional thermoplastic nanofluidic devices with high process yield rates (>90%) and with a short total assembly time (16 min). The approach involves thermally sealing a high T(g) (glass transition temperature) substrate containing the nanofluidic structures to a cover plate possessing a lower T(g). Nanofluidic devices with critical feature sizes ranging between 25-250 nm were fabricated in a thermoplastic substrate (T(g) = 104 °C) and sealed with a cover plate (T(g) = 75 °C) at a temperature significantly below the T(g) of the substrate. Results obtained from sealing tests revealed that the integrity of the nanochannels remained intact after assembly and devices were useful for fluorescence imaging at high signal-to-noise ratios. The functionality of the assembled devices was demonstrated by studying the stretching and translocation dynamics of dsDNA in the enclosed thermoplastic nanofluidic channels.

  1. High Process Yield Rates of Thermoplastic Nanofluidic Devices using a Hybrid Thermal Assembly Technique

    PubMed Central

    Uba, Franklin I.; Hu, Bo; Weerakoon-Ratnayake, Kumuditha; Oliver-Calixte, Nyote; Soper, Steven A.

    2014-01-01

    Over the past decade, thermoplastics have been used as alternative substrates to glass and Si for microfluidic devices because of the diverse and robust fabrication protocols available for thermoplastics that can generate high production rates of the desired structures at low cost and with high replication fidelity, the extensive array of physiochemical properties they possess, and the simple surface activation strategies that can be employed to tune their surface chemistry appropriate for the intended application. While the advantages of polymer microfluidics are currently being realized, the evolution of thermoplastic-based nanofluidic devices is fraught with challenges. One challenge is assembly of the device, which consists of sealing a cover plate to the patterned fluidic substrate. Typically, channel collapse or substrate dissolution occurs during assembly making the device inoperable resulting in low process yield rates. In this work, we report a low temperature hybrid assembly approach for the generation of functional thermoplastic nanofluidic devices with high process yield rates (>90%) with a short total assembly time (16 min). The approach involves thermally sealing a high Tg (glass transition temperature) substrate containing the nanofluidic structures to a cover plate possessing a lower Tg. Nanofluidic devices with critical feature sizes ranging between 25 – 250 nm were fabricated in a thermoplastic substrate (Tg = 104°C) and sealed with a cover plate (Tg = 75°C) at a temperature significantly below the Tg of the substrate. Results obtained from sealing tests revealed that the integrity of the nanochannels remained intact after assembly and devices were useful for fluorescence imaging at high signal-to-noise ratios. The functionality of the assembled devices was demonstrated by studying the stretching and translocation dynamics of dsDNA in the enclosed thermoplastic nanofluidic channels. PMID:25511610

  2. Carbon dots with high fluorescence quantum yield: the fluorescence originates from organic fluorophores

    NASA Astrophysics Data System (ADS)

    Shi, Lei; Yang, Jian Hai; Zeng, Hai Bo; Chen, Yong Mei; Yang, Sheng Chun; Wu, Chao; Zeng, Hao; Yoshihito, Osada; Zhang, Qiqing

    2016-07-01

    In this contribution, we have shown that the organic fluorophores, 5-oxo-3,5-dihydro-2H-thiazolo [3,2-a] pyridine-3,7-dicarboxylic acid (TPDCA) and 5-oxo-3,5-dihydro-2H-thiazolo [3,2-a] pyridine-7-carboxylic acid (TPCA), are the main ingredients and fluorescence origins of N,S-CDs via systematic analyses. It inspires us to deeply analyze and understand the fluorescence origins of carbon dots with high fluorescence quantum yields, which will expand their applications.In this contribution, we have shown that the organic fluorophores, 5-oxo-3,5-dihydro-2H-thiazolo [3,2-a] pyridine-3,7-dicarboxylic acid (TPDCA) and 5-oxo-3,5-dihydro-2H-thiazolo [3,2-a] pyridine-7-carboxylic acid (TPCA), are the main ingredients and fluorescence origins of N,S-CDs via systematic analyses. It inspires us to deeply analyze and understand the fluorescence origins of carbon dots with high fluorescence quantum yields, which will expand their applications. Electronic supplementary information (ESI) available. See DOI: 10.1039/c6nr00451b

  3. OPERATIONAL CHALLENGES IN MIXING AND TRANSFER OF HIGH YIELD STRESS SLUDGE WASTE

    SciTech Connect

    Caldwell, T.; Bhatt, P.

    2009-12-07

    The ability to mobilize and transport non-Newtonian waste is essential to advance the closure of highly radioactive storage tanks. Recent waste removal operations from Tank 12H at the Savannah River Site (SRS) encountered sludge mixtures with a yield stress too high to pump. The waste removal equipment for Tank 12H was designed to mobilize and transport a diluted slurry mixture through an underground 550m long (1800 ft) 0.075m diameter (3 inch) pipeline. The transfer pump was positioned in a well casing submerged in the sludge slurry. The design allowed for mobilized sludge to enter the pump suction while keeping out larger tank debris. Data from a similar tank with known rheological properties were used to size the equipment. However, after installation and startup, field data from Tank 12H confirmed the yield stress of the slurry to exceed 40 Pa, whereas the system is designed for 10 Pa. A revision to the removal strategy was required, which involved metered dilution, blending, and mixing to ensure effective and safe transfer performance. The strategy resulted in the removal of over 255,000 kgs of insoluble solids with four discrete transfer evolutions for a total transfer volume of 2400 m{sup 3} (634,000 gallons) of sludge slurry.

  4. High yielding biomass genotypes of willow (Salix spp.) show differences in below ground biomass allocation

    PubMed Central

    Cunniff, Jennifer; Purdy, Sarah J.; Barraclough, Tim J.P.; Castle, March; Maddison, Anne L.; Jones, Laurence E.; Shield, Ian F.; Gregory, Andrew S.; Karp, Angela

    2015-01-01

    Willows (Salix spp.) grown as short rotation coppice (SRC) are viewed as a sustainable source of biomass with a positive greenhouse gas (GHG) balance due to their potential to fix and accumulate carbon (C) below ground. However, exploiting this potential has been limited by the paucity of data available on below ground biomass allocation and the extent to which it varies between genotypes. Furthermore, it is likely that allocation can be altered considerably by environment. To investigate the role of genotype and environment on allocation, four willow genotypes were grown at two replicated field sites in southeast England and west Wales, UK. Above and below ground biomass was intensively measured over two two-year rotations. Significant genotypic differences in biomass allocation were identified, with below ground allocation differing by up to 10% between genotypes. Importantly, the genotype with the highest below ground biomass also had the highest above ground yield. Furthermore, leaf area was found to be a good predictor of below ground biomass. Growth environment significantly impacted allocation; the willow genotypes grown in west Wales had up to 94% more biomass below ground by the end of the second rotation. A single investigation into fine roots showed the same pattern with double the volume of fine roots present. This greater below ground allocation may be attributed primarily to higher wind speeds, plus differences in humidity and soil characteristics. These results demonstrate that the capacity exists to breed plants with both high yields and high potential for C accumulation. PMID:26339128

  5. Hydraulic conductivity, specific yield, and pumpage--High Plains aquifer system, Nebraska

    USGS Publications Warehouse

    Pettijohn, Robert A.; Chen, Hsiu-Hsiung

    1983-01-01

    Hydrologic data used to evalute the ground-water potential of the High Plains aquifer system in Nebraska are presented on maps showing the hydraulic conductivity and specific yield of the aquifer system and the volume and distribution of water pumped for irrigation from the aquifer system during 1980. The High Plains aquifer system underlies 177,000 square miles in parts of eight states, including 64,770 square miles in Nebraska. It consists of the Ogallala Formation and Tertiary and Quaternary deposits that are saturated and hydraulically connected to the Ogallala. The hydraulic conductivity of the aquifer system varies from greater than 200 feet per day in parts of the North Platte, Platte, Elkhorn, and Republican River valleys to less than 25 feet per day in the northwestern part of the state. Specific yield of the aquifer system ranges from 10 to 20 percent in most of the state and averages 16 percent. The estimated volume of water recoverable from the aquifer system in Nebraska is 2,237 million acre-feet. Inches of water withdrawn from the aquifer system during 1980 varied from less than 1.5 in the sandhills of north-central Nebraska to more than 12 in the Platte River and Blue River basins. This withdrawal represents about 6,703,000 acre-feet of ground water. (USGS)

  6. The cbs mutant strain of Rhodococcus erythropolis KA2-5-1 expresses high levels of Dsz enzymes in the presence of sulfate.

    PubMed

    Tanaka, Yasuhiro; Yoshikawa, Osamu; Maruhashi, Kenji; Kurane, Ryuichiro

    2002-11-01

    Two mutants of the dibenzothiophene-desulfurizing Rhodococcus erythropolis KA2-5-1, strains MS51 and MS316, which express a high level of desulfurizing activity in the presence of sulfate, were isolated using the transposome technique. The level of dibenzothiophene-desulfurization by cell-free extracts prepared from mutants MS51 and MS316 grown on sulfate was about five-fold higher than that by cell-free extracts of the wild-type. This result was consistent with results of Western-blot analysis using antisera specific for DszA, DszB and DszC, the enzymes involved in the desulfurization of dibenzothiophene. Gene analysis of the mutants revealed that the same gene was disrupted in mutants MS51 and MS316 and that the transposon-inserted gene in these strains was the gene for cystathionine beta-synthase, cbs. The cbs mutants also expressed high levels of Dsz enzymes when methionine was used as the sole source of sulfur.

  7. Evolved Streptavidin Mutants Reveal Key Role of Loop Residue in High-affinity Binding

    SciTech Connect

    M Magalhaes; C Melo Czekster; R Guan; V Malashkevich; S Almo; M Levy

    2011-12-31

    We have performed a detailed analysis of streptavidin variants with altered specificity towards desthiobiotin. In addition to changes in key residues which widen the ligand binding pocket and accommodate the more structurally flexible desthiobiotin, the data revealed the role of a key, non-active site mutation at the base of the flexible loop (S52G) which slows dissociation of this ligand by approximately sevenfold. Our data suggest that this mutation results in the loss of a stabilizing contact which keeps this loop open and accessible in the absence of ligand. When this mutation was introduced into the wild-type protein, destabilization of the opened loop conferred a {approx}10-fold decrease in both the on-rate and off-rate for the ligand biotin-4-fluoroscein. A similar effect was observed when this mutation was added to a monomeric form of this protein. Our results provide key insight into the role of the streptavidin flexible loop in ligand binding and maintaining high affinity interactions.

  8. A wide range, high yield and good performance pHEMT switch for MMIC phase shifter

    NASA Astrophysics Data System (ADS)

    Menbari, Babak; Dousti, Massoud; Hajghassem, Hassan

    2014-11-01

    A low-cost method with high yield and good performance is presented by pHEMTs (pseudomorphic high electron mobility transistors) to be used in phase shifter switches. In this method, the capacitor in "off" mode (Coff) of transistor is reduced, without variation of the transistor structure. The transistor structure in switch mode can be optimised. This method increases the transistor isolation in turn "off" mode, while there is no change in resistance of the transistor in "on" mode (Ron). Transistor dimension is determined in turn "off" mode (Vg = -4.5 V) and standard form of 4 × 75 μm. So, in this method, insertion loss will be reduced without a perceptible change in transistor dimension. Thus, design and fabrication capability of some circuits such as phase shifters, antenna switches, SPDT (single port double throw) - without any change in technology - are increasing. In this paper, post layout and measurement result for a sample block of phase shifter are shown.

  9. Reaching High-Yield Fusion with a Slow Plasma Liner Compressing a Magnetized Target

    SciTech Connect

    Ryutov, D D; Parks, P B

    2008-03-18

    Dynamics of the compression of a magnetized plasma target by a heavy liner made of partially ionized high high-Z material is discussed. A 'soft-landing' (shockless) mode of the liner deceleration is analyzed. Conclusion is drawn that such mode is possible for the liners whose thickness at the time of the first contact with the target is smaller than, roughly, 10% of the initial (un-compressed) target radius. A combination of the plasma liner with one or two glide cones allows for a direct access to the area near the center of the reactor chamber. One can then generate plasma target inside the plasma liner at the optimum time. The other advantage of the glide cones is that they can be used to deliver additional fuel to the center of the target near the point of a maximum compression and thereby increase the fusion yield.

  10. High yield recombinant production of a self-assembling polycationic peptide for silica biomineralization.

    PubMed

    Zerfaß, Christian; Braukmann, Sandra; Nietzsche, Sandor; Hobe, Stephan; Paulsen, Harald

    2015-04-01

    We report the recombinant bacterial expression and purification at high yields of a polycationic oligopeptide, P5S3. The sequence of P5S3 was inspired by a diatom silaffin, a silica precipitating peptide. Like its native model, P5S3 exhibits silica biomineralizing activity, but furthermore has unusual self-assembling properties. P5S3 is efficiently expressed in Escherichia coli as fusion with ketosteroid isomerase (KSI), which causes deposition in inclusion bodies. After breaking the fusion by cyanogen bromide reaction, P5S3 was purified by cation exchange chromatography, taking advantage of the exceptionally high content of basic amino acids. The numerous cationic charges do not prevent, but may even promote counterion-independent self-assembly which in turn leads to silica precipitation. Enzymatic phosphorylation, a common modification in native silica biomineralizing peptides, can be used to modify the precipitation activity.

  11. High-Yield Lithium-Injection Fusion-Energy (HYLIFE) reactor

    SciTech Connect

    Blink, J.A.; Hogam, W.J.; Hovingh, J.; Meier, E.R.; Pitts, J.H.

    1985-12-23

    The High-Yield Lithium-Injection Fusion Energy (HYLIFE) concept to convent inertial confinement fusion energy into electric power has undergone intensive research and refinement at LLNL since 1978. This paper reports on the final HYLIFE design, focusing on five major areas: the HYLIFE reaction chamber (which includes neutronics, liquid-metal jet-array hydrocynamics, and structural design), supporting systems, primary steam system and balance of plant, safety and environmental protection, and costs. An annotated bibliography of reports applicable to HYLIFE is also provided. We conclude that HYLIFE is a particularly viable concept for the safe, clean production of electrical energy. The liquid-metal jet array, HYLIFE's key design feature, protects the surrounding structural components from x-rays, fusion fuel-pellet debris, neutron damage and activation, and high temperatures and stresses, allowing the structure to last for the plant's entire 30-year lifetime without being replaced. 127 refs., 18 figs.

  12. High Bone Mass-Causing Mutant LRP5 Receptors Are Resistant to Endogenous Inhibitors In Vivo.

    PubMed

    Niziolek, Paul J; MacDonald, Bryan T; Kedlaya, Rajendra; Zhang, Minjie; Bellido, Teresita; He, Xi; Warman, Matthew L; Robling, Alexander G

    2015-10-01

    Certain missense mutations affecting LRP5 cause high bone mass (HBM) in humans. Based on in vitro evidence, HBM LRP5 receptors are thought to exert their effects by providing resistance to binding/inhibition of secreted LRP5 inhibitors such as sclerostin (SOST) and Dickkopf homolog-1 (DKK1). We previously reported the creation of two Lrp5 HBM knock-in mouse models, in which the human p.A214V or p.G171V missense mutations were knocked into the endogenous Lrp5 locus. To determine whether HBM knock-in mice are resistant to SOST- or DKK1-induced osteopenia, we bred Lrp5 HBM mice with transgenic mice that overexpress human SOST in osteocytes ((8kb) Dmp1-SOST) or mouse DKK1 in osteoblasts and osteocytes ((2.3kb) Col1a1-Dkk1). We observed that the (8kb) Dmp1-SOST transgene significantly lowered whole-body bone mineral density (BMD), bone mineral content (BMC), femoral and vertebral trabecular bone volume fraction (BV/TV), and periosteal bone-formation rate (BFR) in wild-type mice but not in mice with Lrp5 p.G171V and p.A214V alleles. The (2.3kb) Col1a1-Dkk1 transgene significantly lowered whole-body BMD, BMC, and vertebral BV/TV in wild-type mice and affected p.A214V mice more than p.G171V mice. These in vivo data support in vitro studies regarding the mechanism of HBM-causing mutations, and imply that HBM LRP5 receptors differ in their relative sensitivity to inhibition by SOST and DKK1.

  13. High-bone-mass causing mutant LRP5 receptors are resistant to endogenous inhibitors in vivo†

    PubMed Central

    Niziolek, Paul J.; MacDonald, Bryan T.; Kedlaya, Rajendra; Zhang, Minjie; Bellido, Teresita; He, Xi; Warman, Matthew L.; Robling, Alexander G.

    2015-01-01

    Certain missense mutations affecting LRP5 cause high bone mass (HBM) in humans. Based on in vitro evidence, HBM LRP5 receptors are thought to exert their effects by providing resistance to binding/inhibition of secreted LRP5 inhibitors such as sclerostin (SOST) and Dickkopf homolog-1 (DKK1). We previously reported the creation of two Lrp5 HBM knock-in mouse models, in which the human p.A214V or p.G171V missense mutations were knocked into the endogenous Lrp5 locus. To determine whether HBM knock-in mice are resistant to SOST- or DKK1-induced osteopenia, we bred Lrp5 HBM mice with transgenic mice that overexpress human SOST in osteocytes (8kbDmp1-SOST) or mouse DKK1 in osteoblasts and osteocytes (2.3kbCol1a1-Dkk1). We observed that the 8kbDmp1-SOST transgene significantly lowered whole body BMD, BMC, femoral and vertebral BV/TV, and periosteal BFR in wild-type mice but not in mice with Lrp5 p.G171V and p.A214V alleles. The 2.3kbCol1a1-Dkk1 transgene significantly lowered whole body BMD, BMC, and vertebral BV/TV in wild-type mice and affected p.A214V mice more than p.G171V mice. These in vivo data support in vitro studies regarding the mechanism of HBM-causing mutations, and imply that HBM LRP5 receptors differ in their relative sensitivity to inhibition by SOST and DKK1. PMID:25808845

  14. A new genetic method for isolating functionally interacting genes: high plo1(+)-dependent mutants and their suppressors define genes in mitotic and septation pathways in fission yeast.

    PubMed Central

    Cullen, C F; May, K M; Hagan, I M; Glover, D M; Ohkura, H

    2000-01-01

    We describe a general genetic method to identify genes encoding proteins that functionally interact with and/or are good candidates for downstream targets of a particular gene product. The screen identifies mutants whose growth depends on high levels of expression of that gene. We apply this to the plo1(+) gene that encodes a fission yeast homologue of the polo-like kinases. plo1(+) regulates both spindle formation and septation. We have isolated 17 high plo1(+)-dependent (pld) mutants that show defects in mitosis or septation. Three mutants show a mitotic arrest phenotype. Among the 14 pld mutants with septation defects, 12 mapped to known loci: cdc7, cdc15, cdc11 spg1, and sid2. One of the pld mutants, cdc7-PD1, was selected for suppressor analysis. As multicopy suppressors, we isolated four known genes involved in septation in fission yeast: spg1(+), sce3(+), cdc8(+), and rho1(+), and two previously uncharacterized genes, mpd1(+) and mpd2(+). mpd1(+) exhibits high homology to phosphatidylinositol 4-phosphate 5-kinase, while mpd2(+) resembles Saccharomyces cerevisiae SMY2; both proteins are involved in the regulation of actin-mediated processes. As chromosomal suppressors of cdc7-PD1, we isolated mutations of cdc16 that resulted in multiseptation without nuclear division. cdc16(+), dma1(+), byr3(+), byr4(+) and a truncated form of the cdc7 gene were isolated by complementation of one of these cdc16 mutations. These results demonstrate that screening for high dose-dependent mutants and their suppressors is an effective approach to identify functionally interacting genes. PMID:10924454

  15. Hydraulics of high-yield orchard trees: a case study of three Malus domestica cultivars.

    PubMed

    Beikircher, Barbara; De Cesare, Chiara; Mayr, Stefan

    2013-12-01

    The drought tolerance of three economically important apple cultivars, Golden Delicious, Braeburn and Red Delicious, was analysed. The work offers insights into the hydraulics of these high-yield trees and indicates a possible hydraulic limitation of carbon gain. The hydraulic safety and efficiency of branch xylem and leaves were quantified, drought tolerance of living tissues was measured and stomatal regulation, turgor-loss point and osmotic potential at full turgor were analysed. Physiological measurements were correlated with anatomical parameters, such as conduit diameter, cell-wall reinforcement, stomatal density and stomatal pore length. Hydraulic safety differed considerably between the three cultivars with Golden Delicious being significantly less vulnerable to drought-induced embolism than Braeburn and Red Delicious. In Golden Delicious, leaves were less resistant than branch xylem, while in the other cultivars leaves were more resistant than branch xylem. Hydraulic efficiency and xylem anatomical measurements indicate differences in pit properties, which may also be responsible for variations in hydraulic safety. In all three cultivars, full stomatal closure occurred at water potentials where turgor had already been lost and severe loss of hydraulic conductivity as well as damage to living cells had been induced. The consequential negative safety margins pose a risk for hydraulic failure but facilitate carbon gain, which is further improved by the observed high stomatal conductance. Maximal stomatal conductance was clearly seen to be related to stomatal density and size. Based on our results, these three high-yield Malus domestica Borkh. cultivars span a wide range of drought tolerances, appear optimized for maximal carbon gain and, thus, all perform best under well-managed growing conditions. PMID:24319028

  16. Hydraulics of high-yield orchard trees: a case study of three Malus domestica cultivars.

    PubMed

    Beikircher, Barbara; De Cesare, Chiara; Mayr, Stefan

    2013-12-01

    The drought tolerance of three economically important apple cultivars, Golden Delicious, Braeburn and Red Delicious, was analysed. The work offers insights into the hydraulics of these high-yield trees and indicates a possible hydraulic limitation of carbon gain. The hydraulic safety and efficiency of branch xylem and leaves were quantified, drought tolerance of living tissues was measured and stomatal regulation, turgor-loss point and osmotic potential at full turgor were analysed. Physiological measurements were correlated with anatomical parameters, such as conduit diameter, cell-wall reinforcement, stomatal density and stomatal pore length. Hydraulic safety differed considerably between the three cultivars with Golden Delicious being significantly less vulnerable to drought-induced embolism than Braeburn and Red Delicious. In Golden Delicious, leaves were less resistant than branch xylem, while in the other cultivars leaves were more resistant than branch xylem. Hydraulic efficiency and xylem anatomical measurements indicate differences in pit properties, which may also be responsible for variations in hydraulic safety. In all three cultivars, full stomatal closure occurred at water potentials where turgor had already been lost and severe loss of hydraulic conductivity as well as damage to living cells had been induced. The consequential negative safety margins pose a risk for hydraulic failure but facilitate carbon gain, which is further improved by the observed high stomatal conductance. Maximal stomatal conductance was clearly seen to be related to stomatal density and size. Based on our results, these three high-yield Malus domestica Borkh. cultivars span a wide range of drought tolerances, appear optimized for maximal carbon gain and, thus, all perform best under well-managed growing conditions.

  17. Nanoimprinted substrates for high-yield production of topological insulator nanoribbons

    NASA Astrophysics Data System (ADS)

    Wi, Sungjin; Elezi, Eljon; Liu, Amy C.; Ray, Vishva; Sun, Kai; Liang, Xiaogan

    2013-06-01

    We present a growth process mediated by nanoimprinted nanostructures specifically for producing bismuth selenide (Bi2Se3) topological insulator nanoribbons with a high yield. In this process, topological insulator nanostructures are grown on nanoimprinted gratings by using a nanoparticle-catalyzed vapor-liquid-solid mechanism. In comparison with the growth processes performed on flat and randomly rough substrates, such a nanograting-mediated growth method produces topological insulator nanoribbons with a higher yield (˜15 000 nanoribbons/mm2), a narrower average ribbon width ( w avg<60 nm), and a higher uniformity in ribbon width ( σ<30 nm); effectively suppresses the formation of other unwanted morphologies; and also results in the axial growth of nanoribbons along specific in-plane directions relative to pre-structured gratings. Such technical merits of nanograting-mediated growth are attributed to the preferential nucleation of Bi2Se3 crystal seeds and the concomitant pinning of catalytic nanoparticles at ordered grating edges. Finally, Aharonov-Bohm interference oscillations in the magnetoresistance were observed and demonstrated the coherent transport of electrons through topological surface states of Bi2Se3 nanoribbons. This growth process in combination with large-area nanoimprint lithography could serve as an important foundation for nanomanufacturing topological insulator nanoribbons with controllable feature size, large-area uniformity, and ordering, suitable for applications in future low-dissipation nanoelectronics.

  18. Vermicomposting grape marc yields high quality organic biofertiliser and bioactive polyphenols.

    PubMed

    Domínguez, Jorge; Martínez-Cordeiro, Hugo; Álvarez-Casas, Marta; Lores, Marta

    2014-12-01

    Grape is the largest fruit crop in the world, and most (80%) of the harvested fruit is used to make wine. The main by-product of the wine industry is called grape marc, which consists of the stalks, skin, pulp and seeds that remain after pressing the grapes. The aim of this study was to evaluate whether grape marc could be processed by vermicomposting on an industrial scale to yield both a high-quality organic, polyphenol-free fertiliser and grape seeds (as a source of bioactive polyphenols). Vermicomposting reduced the biomass of grape marc substantially (by 58%), mainly as a result of the loss of volatile solids. After 2 weeks, the process yielded a nutrient-rich, microbiologically active and stabilised peat-like material that was easily separated from the seeds by sieving. Although the polyphenol content of the seeds was considerably reduced, this disadvantage was outweighed by the ease of separation of the seeds. Separation of the seeds also eliminated the polyphenol-associated phytotoxicity from the vermicompost. The seeds still contained useful amounts of polyphenols, which could be directly extracted for use in the pharmaceutical, cosmetic and food industries. The procedure described is effective, simple and economical, and could easily be scaled up for industrial application. PMID:25349068

  19. High relative humidity increases yield, harvest index, flowering, and gynophore growth of hydroponically grown peanut plants

    NASA Technical Reports Server (NTRS)

    Mortley, D. G.; Bonsi, C. K.; Loretan, P. A.; Hill, W. A.; Morris, C. E.

    2000-01-01

    Growth chamber experiments were conducted to study the physiological and growth response of peanut (Arachis hypogaea L.) to 50% and 85% relative humidity (RH). The objective was to determine the effects of RH on pod and seed yield, harvest index, and flowering of peanut grown by the nutrient film technique (NFT). 'Georgia Red' peanut plants (14 days old) were planted into growth channels (0.15 x 0.15 x 1.2 m). Plants were spaced 25 cm apart with 15 cm between channels. A modified half-Hoagland solution with an additional 2 mM Ca was used. Solution pH was maintained between 6.4 and 6.7, and electrical conductivity (EC) ranged between 1100 and 1200 microS cm-1. Temperature regimes of 28/22 degrees C were maintained during the light/dark periods (12 hours each) with photosynthetic photon flux (PPF) at canopy level of 500 micromoles-m-2s-1. Foliage and pod fresh and dry weights, total seed yield, harvest index (HI), and seed maturity were greater at high than at low RH. Plants grown at 85% RH had greater total and individual leaflet area and stomatal conductance, flowered 3 days earlier and had a greater number of flowers reaching anthesis. Gynophores grew more rapidly at 85% than at 50% RH.

  20. High relative humidity increases yield, harvest index, flowering, and gynophore growth of hydroponically grown peanut plants.

    PubMed

    Mortley, D G; Bonsi, C K; Loretan, P A; Hill, W A; Morris, C E

    2000-02-01

    Growth chamber experiments were conducted to study the physiological and growth response of peanut (Arachis hypogaea L.) to 50% and 85% relative humidity (RH). The objective was to determine the effects of RH on pod and seed yield, harvest index, and flowering of peanut grown by the nutrient film technique (NFT). 'Georgia Red' peanut plants (14 days old) were planted into growth channels (0.15 x 0.15 x 1.2 m). Plants were spaced 25 cm apart with 15 cm between channels. A modified half-Hoagland solution with an additional 2 mM Ca was used. Solution pH was maintained between 6.4 and 6.7, and electrical conductivity (EC) ranged between 1100 and 1200 microS cm-1. Temperature regimes of 28/22 degrees C were maintained during the light/dark periods (12 hours each) with photosynthetic photon flux (PPF) at canopy level of 500 micromoles-m-2s-1. Foliage and pod fresh and dry weights, total seed yield, harvest index (HI), and seed maturity were greater at high than at low RH. Plants grown at 85% RH had greater total and individual leaflet area and stomatal conductance, flowered 3 days earlier and had a greater number of flowers reaching anthesis. Gynophores grew more rapidly at 85% than at 50% RH.

  1. A rapid method to produce high yields of purified rotavirus particles.

    PubMed

    Villegas, Guillermo Adolfo; Argüelles, Marcelo Horacio; Castello, Alejandro Andrés; Mas, Neus Jiménez; Glikmann, Graciela

    2002-06-01

    A rapid purification method of rotavirus particles to high yield retaining the double shelled structure of infectious virus is described. Group A rotavirus (UK strain) was concentrated through a cushion of colloidal silica (rho=1.10 g/cm(3)) or by precipitating with polyethylene glycol 8000. After concentration, infectious rotavirus was cleared from host cell proteins by density equilibrium centrifugation in gradients of colloidal silica using near vertical rotors. Characterisation of purified virus assessed by electron microscopy and poliacrylamide gel electrophoresis (PAGE) revealed the typical wheel shape structure of rotavirus particles and the presence of the 11 segments of dsRNA arranged in the 4-2-3-2 pattern. Presence of rotavirus structural proteins including VP6, VP4 and VP7 from the outer shell, was demonstrated by SDS-PAGE and Western blot using polyclonal and VP6-specific monoclonal antibodies. This method achieved a approximately 1500 fold purification, which retained approximately 80% infectivity depending on the concentration protocol used, while yielding 160 microg of viral protein per each litre of infected cell culture medium. The time required for the isopycnic centrifugation was only 25 min and the entire completion of the method required 3.5 h. The method is simple technically and applicable to the purification of large as well as minute amounts of virus.

  2. Enhancing biomass energy yield from pilot-scale high rate algal ponds with recycling.

    PubMed

    Park, J B K; Craggs, R J; Shilton, A N

    2013-09-01

    This paper investigates the effect of recycling on biomass energy yield in High Rate Algal Ponds (HRAPs). Two 8 m(3) pilot-scale HRAPs treating primary settled sewage were operated in parallel and monitored over a 2-year period. Volatile suspended solids were measured from both HRAPs and their gravity settlers to determine biomass productivity and harvest efficiency. The energy content of the biomass was also measured. Multiplying biomass productivity and harvest efficiency gives the 'harvestable biomass productivity' and multiplying this by the energy content defines the actual 'biomass energy yield'. In Year 1, algal recycling was implemented in one of the ponds (HRAPr) and improved harvestable biomass productivity by 58% compared with the control (HRAPc) without recycling (HRAPr: 9.2 g/m(2)/d; HRAPc: 5.8 g/m(2)/d). The energy content of the biomass grown in HRAPr, which was dominated by Pediastrun boryanum, was 25% higher than the control HRAPc which contained a mixed culture of 4-5 different algae (HRAPr: 21.5 kJ/g; HRAPc: 18.6 kJ/g). In Year 2, HRAPc was then seeded with the biomass harvested from the P. boryanum dominated HRAPr. This had the effect of shifting algal dominance from 89% Dictyosphaerium sp. (which is poorly-settleable) to over 90% P. boryanum in 5 months. Operation of this pond was then switched to recycling its own harvested biomass, which maintained P. boryanum dominance for the rest of Year 2. This result confirms, for the first time in the literature, that species control is possible for similarly sized co-occurring algal colonies in outdoor HRAP by algal recycling. With regard to the overall improvement in biomass energy yield, which is a critical parameter in the context of algal cultivation for biofuels, the combined improvements that recycling triggered in biomass productivity, harvest efficiency and energy content enhanced the harvested biomass energy yield by 66% (HRAPr: 195 kJ/m(2)/day; HRAPc: 118 kJ/m(2)/day).

  3. A novel, simple, high-throughput method for isolation of genome-wide transposon insertion mutants of Escherichia coli K-12.

    PubMed

    Miki, Takeyoshi; Yamamoto, Yoshihiro; Matsuda, Hideo

    2008-01-01

    We developed a novel, simple, high-throughput method for isolation of genome-wide transposon insertion mutants of Escherichia coli K-12. The basic idea of the method is to randomly disrupt the genes on the DNA fragments cloned on the Kohara library by inserting a mini-transposon first, and then transfer the disrupted genes from the lambda vector to the E. coli chromosome by homologous recombination. Using this method, we constructed a set of 8402 Km(r) cis-diploid mutants harboring a mini-Tn10 insertion mutation and the corresponding wild-type gene on a chromosome, as well as a set of 6954 haploid mutants derived from the cis-diploid mutants. The major advantage of the strategy used is that the indispensable genes or sites for growth can be identified. Preliminary results suggest that 415 open reading frames are indispensable for growth in E. coli cells. A total of 6404 haploid mutants were deposited to Genetic Strains Research Center, National Institute of Genetics, Japan (Chapter 26) and are available for public distribution upon request (http://shigen.lab.nig.ac.jp/ecoli/strain/nbrp/resource.jsp). PMID:18392969

  4. A zebrafish screen for craniofacial mutants identifies wdr68 as a highly conserved gene required for endothelin-1 expression

    PubMed Central

    Nissen, Robert M; Amsterdam, Adam; Hopkins, Nancy

    2006-01-01

    Background Craniofacial birth defects result from defects in cranial neural crest (NC) patterning and morphogenesis. The vertebrate craniofacial skeleton is derived from cranial NC cells and the patterning of these cells occurs within the pharyngeal arches. Substantial efforts have led to the identification of several genes required for craniofacial skeletal development such as the endothelin-1 (edn1) signaling pathway that is required for lower jaw formation. However, many essential genes required for craniofacial development remain to be identified. Results Through screening a collection of insertional zebrafish mutants containing approximately 25% of the genes essential for embryonic development, we present the identification of 15 essential genes that are required for craniofacial development. We identified 3 genes required for hyomandibular development. We also identified zebrafish models for Campomelic Dysplasia and Ehlers-Danlos syndrome. To further demonstrate the utility of this method, we include a characterization of the wdr68 gene. We show that wdr68 acts upstream of the edn1 pathway and is also required for formation of the upper jaw equivalent, the palatoquadrate. We also present evidence that the level of wdr68 activity required for edn1 pathway function differs between the 1st and 2nd arches. Wdr68 interacts with two minibrain-related kinases, Dyrk1a and Dyrk1b, required for embryonic growth and myotube differentiation, respectively. We show that a GFP-Wdr68 fusion protein localizes to the nucleus with Dyrk1a in contrast to an engineered loss of function mutation Wdr68-T284F that no longer accumulated in the cell nucleus and failed to rescue wdr68 mutant animals. Wdr68 homologs appear to exist in all eukaryotic genomes. Notably, we found that the Drosophila wdr68 homolog CG14614 could substitute for the vertebrate wdr68 gene even though insects lack the NC cell lineage. Conclusion This work represents a systematic identification of approximately 25

  5. Tn-seq explorer: a tool for analysis of high-throughput sequencing data of transposon mutant libraries.

    PubMed

    Solaimanpour, Sina; Sarmiento, Felipe; Mrázek, Jan

    2015-01-01

    Tn-seq is a high throughput technique for analysis of transposon mutant libraries. Tn-seq Explorer was developed as a convenient and easy-to-use package of tools for exploration of the Tn-seq data. In a typical application, the user will have obtained a collection of sequence reads adjacent to transposon insertions in a reference genome. The reads are first aligned to the reference genome using one of the tools available for this task. Tn-seq Explorer reads the alignment and the gene annotation, and provides the user with a set of tools to investigate the data and identify possibly essential or advantageous genes as those that contain significantly low counts of transposon insertions. Emphasis is placed on providing flexibility in selecting parameters and methodology most appropriate for each particular dataset. Tn-seq Explorer is written in Java as a menu-driven, stand-alone application. It was tested on Windows, Mac OS, and Linux operating systems. The source code is distributed under the terms of GNU General Public License. The program and the source code are available for download at http://www.cmbl.uga.edu/downloads/programs/Tn_seq_Explorer/ and https://github.com/sina-cb/Tn-seqExplorer.

  6. High-Resolution Crystal Structures of Villin Headpiece nad Mutants with Reduced F-Actin Binding Activity

    SciTech Connect

    Meng,J.; Vardar, D.; Wang, Y.; Guo, H.; Head, J.; McKnight, C.

    2005-01-01

    Villin-type headpiece domains are approximately 70 amino acid modular motifs found at the C terminus of a variety of actin cytoskeleton-associated proteins. The headpiece domain of villin, a protein found in the actin bundles of the brush border epithelium, is of interest both as a compact F-actin binding domain and as a model folded protein. We have determined the high-resolution crystal structures of chicken villin headpiece (HP67) at 1.4 Angstrom resolution as well as two mutants, R37A and W64Y, at 1.45 and 1.5 Angstrom resolution, respectively. Replacement of R37 causes a 5-fold reduction in F-actin binding affinity in sedimentation assays. Replacement of W64 results in a much more drastic reduction in F-actin binding affinity without significant changes in headpiece structure or stability. The detailed comparison of these crystal structures with each other and to our previously determined NMR structures of HP67 and the 35-residue autonomously folding subdomain in villin headpiece, HP35, provides the details of the headpiece fold and further defines the F-actin binding site of villin-type headpiece domains.

  7. Tn-seq explorer: a tool for analysis of high-throughput sequencing data of transposon mutant libraries.

    PubMed

    Solaimanpour, Sina; Sarmiento, Felipe; Mrázek, Jan

    2015-01-01

    Tn-seq is a high throughput technique for analysis of transposon mutant libraries. Tn-seq Explorer was developed as a convenient and easy-to-use package of tools for exploration of the Tn-seq data. In a typical application, the user will have obtained a collection of sequence reads adjacent to transposon insertions in a reference genome. The reads are first aligned to the reference genome using one of the tools available for this task. Tn-seq Explorer reads the alignment and the gene annotation, and provides the user with a set of tools to investigate the data and identify possibly essential or advantageous genes as those that contain significantly low counts of transposon insertions. Emphasis is placed on providing flexibility in selecting parameters and methodology most appropriate for each particular dataset. Tn-seq Explorer is written in Java as a menu-driven, stand-alone application. It was tested on Windows, Mac OS, and Linux operating systems. The source code is distributed under the terms of GNU General Public License. The program and the source code are available for download at http://www.cmbl.uga.edu/downloads/programs/Tn_seq_Explorer/ and https://github.com/sina-cb/Tn-seqExplorer. PMID:25938432

  8. High-yield electrosynthesis of hydrogen peroxide from oxygen reduction by hierarchically porous carbon.

    PubMed

    Liu, Yanming; Quan, Xie; Fan, Xinfei; Wang, Hua; Chen, Shuo

    2015-06-01

    H2O2 production by electroreduction of O2 is an attractive alternative to the current anthraquinone process, which is highly desirable for chemical industries and environmental remediation. However, it remains a great challenge to develop cost-effective electrocatalysts for H2O2 synthesis. Here, hierarchically porous carbon (HPC) was proposed for the electrosynthesis of H2O2 from O2 reduction. It exhibited high activity for O2 reduction and good H2O2 selectivity (95.0-70.2%, most of them >90.0% at pH 1-4 and >80.0% at pH 7). High-yield H2O2 generation has been achieved on HPC with H2O2 concentrations of 222.6-62.0 mmol L(-1) (2.5 h) and corresponding H2O2 production rates of 395.7-110.2 mmol h(-1)  g(-1) at pH 1-7 and -0.5 V. Moreover, HPC was energy-efficient for H2O2 production with current efficiency of 81.8-70.8%. The exceptional performance of HPC for electrosynthesis of H2O2 could be attributed to its high content of sp(3)-C and defects, large surface area and fast mass transfer. PMID:25892325

  9. Garnet Yield Strength at High Pressures and Implications for Upper Mantle and Transition Zone Rheology

    SciTech Connect

    Kavner,A.

    2008-01-01

    Garnet helps control the mechanical behavior of the Earth's crust, mantle, and transition zone. Here, measurements are presented suggesting that garnet, long considered to be a high-viscosity phase, is actually weaker than the other dominant components in the transition zone. The mechanical behavior of garnet at high pressures was examined using radial diffraction techniques in the diamond anvil cell. The yield strength of grossular garnet was inferred from synchrotron X-ray measurements of differential lattice strains. The differential stress was found to increase from 1.3 ({+-}0.6) GPa at a hydrostatic pressure 5.8 ({+-}1.1) GPa to 4.1 ({+-}0.4) GPa at 15.7 ({+-}1.0) GPa, where it was level to 19 GPa. The strength results are consistent with inferred strength values for majorite garnet from measurements in the diamond cell normal geometry, bolstering the idea that garnet-structured materials may all have similar strengths. In this low-temperature, high differential stress regime, garnet is shown to be significantly weaker than anhydrous ringwoodite and to have a strength similar to hydrous ringwoodite. This result suggests that the presence of water in the transition zone may not be required to explain a weak rheology, and therefore models of transition zone behavior built assuming that garnet is the high-strength phase may need to be revised.

  10. Synthesis of uniform CdS nanowires in high yield and its single nanowire electrical property

    SciTech Connect

    Yan Shancheng; Sun Litao; Qu Peng; Huang Ninping; Song Yinchen; Xiao Zhongdang

    2009-10-15

    Large-scale high quality CdS nanowires with uniform diameter were synthesized by using a rapid and simple solvothermal route. Field emission scan electron microscopy (FESEM) and transmission electron microscopy (TEM) images show that the CdS nanowires have diameter of about 26 nm and length up to several micrometres. High resolution TEM (HRTEM) study indicates the single-crystalline nature of CdS nanowires with an oriented growth along the c-axis direction. The optical properties of the products were characterized by UV-vis absorption spectra, photoluminescence spectra and Raman spectra. The resistivity, electron concentration and electron mobility of single NW are calculated by fitting the symmetric I-V curves measured on single NW by the metal-semiconductor-metal model based on thermionic field emission theory. - Graphical abstract: Large-scale high quality CdS nanowires (NWs) with uniform diameter were synthesized by using a rapid and simple solvothermal route. The reaction time is reduced to 2 h, comparing to other synthesis which needed long reaction time up to 12 h. In addition, the as-prepared CdS nanowires have more uniform diameter and high yield. More importantly, the I-V curve of present single CdS nanowire has a good symmetric characteristic as expected by the theory.

  11. High-yield synthesis and optical properties of g-C3N4.

    PubMed

    Yuan, Yanwen; Zhang, Lulu; Xing, Jun; Utama, M Iqbal Bakti; Lu, Xin; Du, Kezhao; Li, Yongmei; Hu, Xiao; Wang, Shijie; Genç, Aziz; Dunin-Borkowski, Rafal; Arbiol, Jordi; Xiong, Qihua

    2015-08-01

    Graphitic carbon nitride (g-C3N4), a metal-free semiconductor with a band gap of 2.7 eV, has received considerable attention owing to its fascinating photocatalytic performances under visible-light. g-C3N4 exhibits high thermal and chemical stability and non-toxicity such that it has been considered as the most promising photocatalyst for environmental improvement and energy conservation. Hence, it is of great importance to obtain high-quality g-C3N4 and gain a clear understanding of its optical properties. Herein, we report a high-yield synthesis of g-C3N4 products via heating of high vacuum-sealed melamine powder in an ampoule at temperatures between 450 and 650 °C. Using transmission electron microscopy (TEM), scanning transmission electron microscopy (STEM), electron energy loss spectroscopy (EELS), thermogravimetric analysis (TGA), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS), the chemical composition and crystallization of the as-produced g-C3N4 are demonstrated. A systematic optical study of g-C3N4 is carried out with several approaches. The optical phonon behavior of g-C3N4 is revealed by infrared and Raman spectroscopy, and the emission properties of g-C3N4 are investigated using photoluminescence (PL) spectroscopy, while the photocatalytic properties are explored by the photodegradation experiment. PMID:26152840

  12. A Scalable and High-Yield Strategy for the Synthesis of Sequence-Defined Macromolecules.

    PubMed

    Solleder, Susanne C; Zengel, Deniz; Wetzel, Katharina S; Meier, Michael A R

    2016-01-18

    The efficient synthesis of a sequence-defined decamer, its characterization, and its straightforward dimerization through self-metathesis are described. For this purpose, a monoprotected AB monomer was designed and used to synthesize a decamer bearing ten different and selectable side chains by iterative Passerini three-component reaction (P-3CR) and subsequent deprotection. The highly efficient procedure provided excellent yields and allows for the multigram-scale synthesis of such perfectly defined macromolecules. An olefin was introduced at the end of the synthesis, allowing the self-metathesis reaction of the resulting decamer to provide a sequence-defined 20-mer with a molecular weight of 7046.40 g mol(-1). The obtained oligomers were carefully characterized by NMR and IR spectroscopy, GPC and GPC coupled to ESI-MS, and mass spectrometry (FAB and orbitrap ESI-MS).

  13. A high liquid yield process for retorting various organic materials including oil shale

    DOEpatents

    Coburn, T.T.

    1988-07-26

    This invention is a continuous retorting process for various high molecular weight organic materials, including oil shale, that yields an enhanced output of liquid product. The organic material, mineral matter, and an acidic catalyst, that appreciably adsorbs alkenes on surface sites at prescribed temperatures, are mixed and introduced into a pyrolyzer. A circulating stream of olefin enriched pyrolysis gas is continuously swept through the organic material and catalyst, whereupon, as the result of pyrolysis, the enhanced liquid product output is provided. Mixed spent organic material, mineral matter, and cool catalyst are continuously withdrawn from the pyrolyzer. Combustion of the spent organic material and mineral matter serves to reheat the catalyst. Olefin depleted pyrolysis gas, from the pyrolyzer, is enriched in olefins and recycled into the pyrolyzer. The reheated acidic catalyst is separated from the mineral matter and again mixed with fresh organic material, to maintain the continuously cyclic process. 2 figs.

  14. High liquid yield process for retorting various organic materials including oil shale

    DOEpatents

    Coburn, Thomas T.

    1990-01-01

    This invention is a continuous retorting process for various high molecular weight organic materials, including oil shale, that yields an enhanced output of liquid product. The organic material, mineral matter, and an acidic catalyst, that appreciably adsorbs alkenes on surface sites at prescribed temperatures, are mixed and introduced into a pyrolyzer. A circulating stream of olefin enriched pyrolysis gas is continuously swept through the organic material and catalyst, whereupon, as the result of pyrolysis, the enhanced liquid product output is provided. Mixed spent organic material, mineral matter, and cool catalyst are continuously withdrawn from the pyrolyzer. Combustion of the spent organic material and mineral matter serves to reheat the catalyst. Olefin depleted pyrolysis gas, from the pyrolyzer, is enriched in olefins and recycled into the pyrolyzer. The reheated acidic catalyst is separated from the mineral matter and again mixed with fresh organic material, to maintain the continuously cyclic process.

  15. A Scalable and High-Yield Strategy for the Synthesis of Sequence-Defined Macromolecules.

    PubMed

    Solleder, Susanne C; Zengel, Deniz; Wetzel, Katharina S; Meier, Michael A R

    2016-01-18

    The efficient synthesis of a sequence-defined decamer, its characterization, and its straightforward dimerization through self-metathesis are described. For this purpose, a monoprotected AB monomer was designed and used to synthesize a decamer bearing ten different and selectable side chains by iterative Passerini three-component reaction (P-3CR) and subsequent deprotection. The highly efficient procedure provided excellent yields and allows for the multigram-scale synthesis of such perfectly defined macromolecules. An olefin was introduced at the end of the synthesis, allowing the self-metathesis reaction of the resulting decamer to provide a sequence-defined 20-mer with a molecular weight of 7046.40 g mol(-1). The obtained oligomers were carefully characterized by NMR and IR spectroscopy, GPC and GPC coupled to ESI-MS, and mass spectrometry (FAB and orbitrap ESI-MS). PMID:26663541

  16. A quartz nanopillar hemocytometer for high-yield separation and counting of CD4+ T lymphocytes

    NASA Astrophysics Data System (ADS)

    Kim, Dong-Joo; Seol, Jin-Kyeong; Wu, Yu; Ji, Seungmuk; Kim, Gil-Sung; Hyung, Jung-Hwan; Lee, Seung-Yong; Lim, Hyuneui; Fan, Rong; Lee, Sang-Kwon

    2012-03-01

    We report the development of a novel quartz nanopillar (QNP) array cell separation system capable of selectively capturing and isolating a single cell population including primary CD4+ T lymphocytes from the whole pool of splenocytes. Integrated with a photolithographically patterned hemocytometer structure, the streptavidin (STR)-functionalized-QNP (STR-QNP) arrays allow for direct quantitation of captured cells using high content imaging. This technology exhibits an excellent separation yield (efficiency) of ~95.3 +/- 1.1% for the CD4+ T lymphocytes from the mouse splenocyte suspensions and good linear response for quantitating captured CD4+ T-lymphoblasts, which is comparable to flow cytometry and outperforms any non-nanostructured surface capture techniques, i.e. cell panning. This nanopillar hemocytometer represents a simple, yet efficient cell capture and counting technology and may find immediate applications for diagnosis and immune monitoring in the point-of-care setting.We report the development of a novel quartz nanopillar (QNP) array cell separation system capable of selectively capturing and isolating a single cell population including primary CD4+ T lymphocytes from the whole pool of splenocytes. Integrated with a photolithographically patterned hemocytometer structure, the streptavidin (STR)-functionalized-QNP (STR-QNP) arrays allow for direct quantitation of captured cells using high content imaging. This technology exhibits an excellent separation yield (efficiency) of ~95.3 +/- 1.1% for the CD4+ T lymphocytes from the mouse splenocyte suspensions and good linear response for quantitating captured CD4+ T-lymphoblasts, which is comparable to flow cytometry and outperforms any non-nanostructured surface capture techniques, i.e. cell panning. This nanopillar hemocytometer represents a simple, yet efficient cell capture and counting technology and may find immediate applications for diagnosis and immune monitoring in the point-of-care setting

  17. A high-yield sampler for toxicological characterization of complex mixtures in combustion effluents.

    PubMed Central

    Kruzel, E L; Lafleur, A L; Braun, A G; Longwell, J P; Thilly, W G; Peters, W A

    1991-01-01

    Combustion sampling for toxicological assessment often requires that large (greater than 100 mg) lots of complex organic mixtures of wide volatility range be rapidly recovered from high temperature gases without contamination. A new sampler, meeting these criteria for studies of public health interest, has been developed and demonstrated. The device provides high sampling rates and intimate contacting of the samples stream with large volumes of a well-cooled, liquid solvent, dichloromethane (DCM). This promotes rapid organics dissolution from carrier gas and particulates and prompt dilution and quenching of the resulting solution, resulting in high organics collection efficiencies with minimal DCM losses. Solvent separation then remits large quantities of concentrated organics for chemical analysis and toxicological testing. One- to seven-hour interrogations of in-flame, post-flame, and flue gas regions gave 50- to 250-mg yields of complex organic mixtures. In side-by-side sampling of combustion exhaust, the DCM sampler provided higher yields of DCM solubles (identified with complex organic mixtures) and of S. typhimuirim mutagens (active without exogenous metabolizing agents) than did a filter/polymeric sorbent bed sampling train. The new sampler also collects polar and high volatile hydrocarbons such as benzaheyde, pentadiyne, m- and p-diethynyl-benzene, and 1-hexen-3,5-diyne. Nitration of naphthalene and pyrene in DCM solution (1 mg/mL each) was less than 1 part in 10(7) after a 345-min exposure to a bubbling flow of moist N2/air mixture (1:1 v/v) containing 107 ppm NO and 1.5 ppm NO2, indicating that for these condition a DCM sampler should resist artifactual nitration of aromatics. However, because of the very high bacterial mutagenicity of some nitroaromatics and the wide range of sampling conditions of environmental interest, nitration and all artifacts must still be scrutinized when using the DCM sampler. The DCM sampler is expected to contribute to public

  18. Study of a High-Yield Cellulase System Created by Heavy-Ion Irradiation-Induced Mutagenesis of Aspergillus niger and Mixed Fermentation with Trichoderma reesei.

    PubMed

    Wang, Shu-Yang; Jiang, Bo-Ling; Zhou, Xiang; Chen, Ji-Hong; Li, Wen-Jian; Liu, Jing; Hu, Wei; Xiao, Guo-Qing; Dong, Miao-Yin; Wang, Yu-Chen

    2015-01-01

    The aim of this study was to evaluate and validate the efficiency of 12C6+ irradiation of Aspergillus niger (A. niger) or mutagenesis via mixed Trichoderma viride (T. viride) culturing as well as a liquid cultivation method for cellulase production via mixed Trichoderma reesei (T. reesei) and A. niger culture fermentation. The first mutagenesis approach was employed to optimize yield from a cellulase-producing strain via heavy-ion mutagenesis and high-throughput screening, and the second was to effectively achieve enzymatic hydrolysis of cellulase from a mixed culture of mutant T. viride and A. niger. We found that 12C6+-ion irradiation induced changes in cellulase biosynthesis in A. niger but had no effect on the time course of the synthesis. It is notable that the exoglucanases (CBH) activities of A. niger strains H11-1 and H differed (6.71 U/mL vs. 6.01 U/mL) and were significantly higher than that of A. niger mutant H3-1. Compared with strain H, the filter paper assay (FPA), endoglucanase (EG) and β-glucosidase (BGL) activities of mutant strain H11-1 were increased by 250.26%, 30.26% and 34.91%, respectively. A mixed culture system was successfully optimized, and the best ratio of T. reesei to A. niger was 5:1 for 96 h with simultaneous inoculation. The BGL activity of the mixed culture increased after 72 h. At 96 h, the FPA and BGL activities of the mixed culture were 689.00 and 797.15 U/mL, respectively, significantly higher than those of monocultures, which were 408.70 and 646.98 U/mL for T. reesei and 447.29 and 658.89 U/mL for A. niger, respectively. The EG activity of the mixed culture was 2342.81 U/mL, a value that was significantly higher than that of monocultures at 2206.57 U/mL for T. reesei and 1727.62 U/mL for A. niger. In summary, cellulose production and hydrolysis yields were significantly enhanced by the proposed combination scheme.

  19. Study of a High-Yield Cellulase System Created by Heavy-Ion Irradiation-Induced Mutagenesis of Aspergillus niger and Mixed Fermentation with Trichoderma reesei.

    PubMed

    Wang, Shu-Yang; Jiang, Bo-Ling; Zhou, Xiang; Chen, Ji-Hong; Li, Wen-Jian; Liu, Jing; Hu, Wei; Xiao, Guo-Qing; Dong, Miao-Yin; Wang, Yu-Chen

    2015-01-01

    The aim of this study was to evaluate and validate the efficiency of 12C6+ irradiation of Aspergillus niger (A. niger) or mutagenesis via mixed Trichoderma viride (T. viride) culturing as well as a liquid cultivation method for cellulase production via mixed Trichoderma reesei (T. reesei) and A. niger culture fermentation. The first mutagenesis approach was employed to optimize yield from a cellulase-producing strain via heavy-ion mutagenesis and high-throughput screening, and the second was to effectively achieve enzymatic hydrolysis of cellulase from a mixed culture of mutant T. viride and A. niger. We found that 12C6+-ion irradiation induced changes in cellulase biosynthesis in A. niger but had no effect on the time course of the synthesis. It is notable that the exoglucanases (CBH) activities of A. niger strains H11-1 and H differed (6.71 U/mL vs. 6.01 U/mL) and were significantly higher than that of A. niger mutant H3-1. Compared with strain H, the filter paper assay (FPA), endoglucanase (EG) and β-glucosidase (BGL) activities of mutant strain H11-1 were increased by 250.26%, 30.26% and 34.91%, respectively. A mixed culture system was successfully optimized, and the best ratio of T. reesei to A. niger was 5:1 for 96 h with simultaneous inoculation. The BGL activity of the mixed culture increased after 72 h. At 96 h, the FPA and BGL activities of the mixed culture were 689.00 and 797.15 U/mL, respectively, significantly higher than those of monocultures, which were 408.70 and 646.98 U/mL for T. reesei and 447.29 and 658.89 U/mL for A. niger, respectively. The EG activity of the mixed culture was 2342.81 U/mL, a value that was significantly higher than that of monocultures at 2206.57 U/mL for T. reesei and 1727.62 U/mL for A. niger. In summary, cellulose production and hydrolysis yields were significantly enhanced by the proposed combination scheme. PMID:26656155

  20. Study of a High-Yield Cellulase System Created by Heavy-Ion Irradiation-Induced Mutagenesis of Aspergillus niger and Mixed Fermentation with Trichoderma reesei

    PubMed Central

    Chen, Ji-Hong; Li, Wen-Jian; Liu, Jing; Hu, Wei; Xiao, Guo-Qing; Dong, Miao-Yin; Wang, Yu-Chen

    2015-01-01

    The aim of this study was to evaluate and validate the efficiency of 12C6+ irradiation of Aspergillus niger (A. niger) or mutagenesis via mixed Trichoderma viride (T. viride) culturing as well as a liquid cultivation method for cellulase production via mixed Trichoderma reesei (T. reesei) and A. niger culture fermentation. The first mutagenesis approach was employed to optimize yield from a cellulase-producing strain via heavy-ion mutagenesis and high-throughput screening, and the second was to effectively achieve enzymatic hydrolysis of cellulase from a mixed culture of mutant T. viride and A. niger. We found that 12C6+-ion irradiation induced changes in cellulase biosynthesis in A. niger but had no effect on the time course of the synthesis. It is notable that the exoglucanases (CBH) activities of A. niger strains H11-1 and H differed (6.71 U/mL vs. 6.01 U/mL) and were significantly higher than that of A. niger mutant H3-1. Compared with strain H, the filter paper assay (FPA), endoglucanase (EG) and β-glucosidase (BGL) activities of mutant strain H11-1 were increased by 250.26%, 30.26% and 34.91%, respectively. A mixed culture system was successfully optimized, and the best ratio of T. reesei to A. niger was 5:1 for 96 h with simultaneous inoculation. The BGL activity of the mixed culture increased after 72 h. At 96 h, the FPA and BGL activities of the mixed culture were 689.00 and 797.15 U/mL, respectively, significantly higher than those of monocultures, which were 408.70 and 646.98 U/mL for T. reesei and 447.29 and 658.89 U/mL for A. niger, respectively. The EG activity of the mixed culture was 2342.81 U/mL, a value that was significantly higher than that of monocultures at 2206.57 U/mL for T. reesei and 1727.62 U/mL for A. niger. In summary, cellulose production and hydrolysis yields were significantly enhanced by the proposed combination scheme. PMID:26656155

  1. RNA mutagenesis yields highly diverse mRNA libraries for in vitro protein evolution

    PubMed Central

    Kopsidas, George; Carman, Rachael K; Stutt, Emma L; Raicevic, Anna; Roberts, Anthony S; Siomos, Mary-Anne V; Dobric, Nada; Pontes-Braz, Luisa; Coia, Greg

    2007-01-01

    Background In protein drug development, in vitro molecular optimization or protein maturation can be used to modify protein properties. One basic approach to protein maturation is the introduction of random DNA mutations into the target gene sequence to produce a library of variants that can be screened for the preferred protein properties. Unfortunately, the capability of this approach has been restricted by deficiencies in the methods currently available for random DNA mutagenesis and library generation. Current DNA based methodologies generally suffer from nucleotide substitution bias that preferentially mutate particular base pairs or show significant bias with respect to transitions or transversions. In this report, we describe a novel RNA-based random mutagenesis strategy that utilizes Qβ replicase to manufacture complex mRNA libraries with a mutational spectrum that is close to the ideal. Results We show that Qβ replicase generates all possible base substitutions with an equivalent preference for mutating A/T or G/C bases and with no significant bias for transitions over transversions. To demonstrate the high diversity that can be sampled from a Qβ replicase-generated mRNA library, the approach was used to evolve the binding affinity of a single domain VNAR shark antibody fragment (12Y-2) against malarial apical membrane antigen-1 (AMA-1) via ribosome display. The binding constant (KD) of 12Y-2 was increased by 22-fold following two consecutive but discrete rounds of mutagenesis and selection. The mutagenesis method was also used to alter the substrate specificity of β-lactamase which does not significantly hydrolyse the antibiotic cefotaxime. Two cycles of RNA mutagenesis and selection on increasing concentrations of cefotaxime resulted in mutants with a minimum 10,000-fold increase in resistance, an outcome achieved faster and with fewer overall mutations than in comparable studies using other mutagenesis strategies. Conclusion The RNA based approach

  2. Microwave absorption properties of amorphous iron nanostructures fabricated by a high-yield method

    NASA Astrophysics Data System (ADS)

    Wang, Zhen; Zuo, Yalu; Yao, Yuelin; Xi, Li; Du, Jihong; Wang, Jianbo; Xue, Desheng

    2013-04-01

    Amorphous Fe nanoparticles and a nanonecklace were synthesized at room temperature by an aqueous reduction procedure, which provided a simple and potential method for volume production of ferromagnetic materials. The morphology was examined by scanning electron microscopy and transmission electron microscopy. The amorphism of Fe nanoparticles and the nanonecklace was confirmed by x-ray diffraction and electron diffraction patterns in transmission electron microscopy. The complex permittivity and permeability behaviour of amorphous iron nanoparticles/paraffin wax (NPPW) and nanonecklace/paraffin wax (NCPW) composites was investigated in 0.1-18 GHz by a coaxial method. The strongest reflection loss values of NPPW and NCPW calculated from permittivity and permeability reached -53.2 dB and -47.8 dB at 6.4 GHz and 4.6 GHz with matching thicknesses of 2.4 mm and 2.3 mm, respectively. Moreover, the frequency ranges of microwave absorption exceeding 90% were around 4.9-8.8 GHz and 3.7-6.1 GHz for NPPW and NCPW, respectively. Comparing the microwave absorption property with crystallized Fe nanostructures, we may conclude that the relatively high resistivity and low permittivity of amorphous Fe nanostructures are favourable for impedance matching, and consequently result in the attracting microwave absorption property of amorphous Fe nanostructures. Thus, amorphous iron nanoparticles and the nanonecklace prepared by a high-yield method have great potential to be a highly efficient microwave absorber.

  3. Mid-Pliocene warm-period deposits in the High Arctic yield insight into camel evolution

    PubMed Central

    Rybczynski, Natalia; Gosse, John C.; Richard Harington, C.; Wogelius, Roy A.; Hidy, Alan J.; Buckley, Mike

    2013-01-01

    The mid-Pliocene was a global warm period, preceding the onset of Quaternary glaciations. Here we use cosmogenic nuclide dating to show that a fossiliferous terrestrial deposit that includes subfossil trees and the northern-most evidence of Pliocene ice wedge casts in Canada’s High Arctic (Ellesmere Island, Nunavut) was deposited during the mid-Pliocene warm period. The age estimates correspond to a general maximum in high latitude mean winter season insolation, consistent with the presence of a rich, boreal-type forest. Moreover, we report that these deposits have yielded the first evidence of a High Arctic camel, identified using collagen fingerprinting of a fragmentary fossil limb bone. Camels originated in North America and dispersed to Eurasia via the Bering Isthmus, an ephemeral land bridge linking Alaska and Russia. The results suggest that the evolutionary history of modern camels can be traced back to a lineage of giant camels that was well established in a forested Arctic. PMID:23462993

  4. High-yielding one-pot synthesis of diaryliodonium triflates from arenes and iodine or aryl iodides.

    PubMed

    Bielawski, Marcin; Olofsson, Berit

    2007-06-28

    Unsymmetric and symmetric diaryliodonium triflates are synthesized from both electron-deficient and electron-rich substrates in a fast, high yielding, and operationally simple protocol employing arenes and aryl iodides or iodine.

  5. HIGH YIELD SOLUBLE BACTERIAL EXPRESSION AND STREAMLINED PURIFICATION OF RECOMBINANT HUMAN INTERFERON α-2A

    PubMed Central

    Bis, Regina L.; Stauffer, Tara M.; Singh, Surinder M.; Lavoie, Thomas B.; Mallela, Krishna M.G.

    2014-01-01

    Interferon α-2a (IFNA2) is a member of the Type I interferon cytokine family, known for its antiviral and anti-proliferative functions. The role of this family in the innate immune response makes it an attractive candidate for the treatment of many viral and chronic immune-compromised diseases. Recombinant IFNA2 is clinically used to modulate hairy cell leukemia as well as hepatitis c. Historically, IFNA2 has been purified from human leukocytes as well as bacterial expression systems. In most cases, bacterial expression of IFNA2 resulted in inclusion body formation, or required numerous purification steps that decreased the protein yield. Here, we describe an expression and purification scheme for IFNA2 using a pET-SUMO bacterial expression system and a single purification step. Using SUMO protein as the fusion tag achieved high soluble protein expression. The SUMO tag was cleaved with the Ulp1 protease leaving no additional amino acids on the fusion terminus following the cleavage. Mass spectrometry, circular dichroism, 2D heteronuclear NMR, and analytical ultracentrifugation confirmed the amino acid sequence identity, secondary and tertiary protein structures, and the solution behavior of the purified IFNA2. The purified protein also has antiviral and anti-proliferative activities comparable to the WHO International Standard, NIBSC 95/650, and the IFNA2 standard available from PBL Assay Science. Combining the expression and purification protocols developed here to produce IFNA2 on a laboratory scale with the commercial fermenter technology commonly used in pharmaceutical industry may further enhance IFNA2 yields, which will promote the development of interferon-based protein drugs to treat various disorders. PMID:24794500

  6. Evaluation of High-Yield Purification Methods on Monodisperse PEG-Grafted Iron Oxide Nanoparticles.

    PubMed

    Lassenberger, Andrea; Bixner, Oliver; Gruenewald, Tilman; Lichtenegger, Helga; Zirbs, Ronald; Reimhult, Erik

    2016-05-01

    Fundamental research on nanoparticle (NP) interactions and development of next-generation biomedical NP applications relies on synthesis of monodisperse, functional, core-shell nanoparticles free of residual dispersants with truly homogeneous and controlled physical properties. Still, synthesis and purification of e.g. such superparamagnetic iron oxide NPs remain a challenge. Comparing the success of different methods is marred by the sensitivity of analysis methods to the purity of the product. We synthesize monodisperse, oleic acid (OA)-capped, Fe3O4 NPs in the superparamagnetic size range (3-10 nm). Ligand exchange of OA for poly(ethylene glycol) (PEG) was performed with the PEG irreversibly grafted to the NP surface by a nitrodopamine (NDA) anchor. Four different methods were investigated to remove excess ligands and residual OA: membrane centrifugation, dialysis, size exclusion chromatography, and precipitation combined with magnetic decantation. Infrared spectroscopy and thermogravimetric analysis were used to determine the purity of samples after each purification step. Importantly, only magnetic decantation yielded pure NPs at high yields with sufficient grafting density for biomedical applications (∼1 NDA-PEG(5 kDa)/nm(2), irrespective of size). The purified NPs withstand challenging tests such as temperature cycling in serum and long-term storage in biological buffers. Dynamic light scattering, transmission electron microscopy, and small-angle X-ray scattering show stability over at least 4 months also in serum. The successful synthesis and purification route is compatible with any conceivable functionalization for biomedical or biomaterial applications of PEGylated Fe3O4 NPs. PMID:27046133

  7. Evaluation of High-Yield Purification Methods on Monodisperse PEG-Grafted Iron Oxide Nanoparticles

    PubMed Central

    2016-01-01

    Fundamental research on nanoparticle (NP) interactions and development of next-generation biomedical NP applications relies on synthesis of monodisperse, functional, core–shell nanoparticles free of residual dispersants with truly homogeneous and controlled physical properties. Still, synthesis and purification of e.g. such superparamagnetic iron oxide NPs remain a challenge. Comparing the success of different methods is marred by the sensitivity of analysis methods to the purity of the product. We synthesize monodisperse, oleic acid (OA)-capped, Fe3O4 NPs in the superparamagnetic size range (3–10 nm). Ligand exchange of OA for poly(ethylene glycol) (PEG) was performed with the PEG irreversibly grafted to the NP surface by a nitrodopamine (NDA) anchor. Four different methods were investigated to remove excess ligands and residual OA: membrane centrifugation, dialysis, size exclusion chromatography, and precipitation combined with magnetic decantation. Infrared spectroscopy and thermogravimetric analysis were used to determine the purity of samples after each purification step. Importantly, only magnetic decantation yielded pure NPs at high yields with sufficient grafting density for biomedical applications (∼1 NDA-PEG(5 kDa)/nm2, irrespective of size). The purified NPs withstand challenging tests such as temperature cycling in serum and long-term storage in biological buffers. Dynamic light scattering, transmission electron microscopy, and small-angle X-ray scattering show stability over at least 4 months also in serum. The successful synthesis and purification route is compatible with any conceivable functionalization for biomedical or biomaterial applications of PEGylated Fe3O4 NPs. PMID:27046133

  8. Characterization of mutants of a highly cross-reactive calcium-binding protein from Brassica pollen for allergen-specific immunotherapy.

    PubMed

    Garmatiuk, Tetiana; Swoboda, Ines; Twardosz-Kropfmüller, Anna; Dall'antonia, Fabio; Keller, Walter; Singh, Mohan B; Bhalla, Prem L; Okada, Takashi; Toriyama, Kinya; Weber, Milena; Ghannadan, Minoo; Sperr, Wolfgang R; Blatt, Katharina; Valent, Peter; Klein, Brigitte; Niederberger, Verena; Curin, Mirela; Balic, Nadja; Spitzauer, Susanne; Valenta, Rudolf

    2013-09-01

    The major turnip (Brassica rapa) pollen allergen, belongs to a family of calcium-binding proteins (i.e., two EF-hand proteins), which occur as highly cross-reactive allergens in pollen of weeds, grasses and trees. In this study, the IgE binding capacity and allergenic activity of three recombinant allergen variants containing mutations in their calcium-binding sites were analyzed in sensitized patients with the aim to identify the most suitable hypoallergenic molecule for specific immunotherapy. Analysis of the wildtype allergen and the mutants regarding IgE reactivity and activation of basophils in allergic patients indicated that the allergen derivative mutated in both calcium-binding domains had the lowest allergenic activity. Gel filtration and circular dichroism experiments showed that both, the wildtype and the double mutant, occurred as dimers in solution and assumed alpha-helical fold, respectively. However, both fold and thermal stability were considerably reduced in the double mutant. The use of bioinformatic tools for evaluation of the solvent accessibility and charge distribution suggested that the reduced IgE reactivity and different structural properties of the double mutant may be due to a loss of negatively charged amino acids on the surface. Interestingly, immunization of rabbits showed that only the double mutant but not the wildtype allergen induced IgG antibodies which recognized the allergen and blocked binding of allergic patients IgE. Due to the extensive structural similarity and cross-reactivity between calcium-binding pollen allergens the hypoallergenic double mutant may be useful not only for immunotherapy of turnip pollen allergy, but also for the treatment of allergies to other two EF-hand pollen allergens. PMID:23790497

  9. Characterization of mutants of a highly cross-reactive calcium-binding protein from Brassica pollen for allergen-specific immunotherapy.

    PubMed

    Garmatiuk, Tetiana; Swoboda, Ines; Twardosz-Kropfmüller, Anna; Dall'antonia, Fabio; Keller, Walter; Singh, Mohan B; Bhalla, Prem L; Okada, Takashi; Toriyama, Kinya; Weber, Milena; Ghannadan, Minoo; Sperr, Wolfgang R; Blatt, Katharina; Valent, Peter; Klein, Brigitte; Niederberger, Verena; Curin, Mirela; Balic, Nadja; Spitzauer, Susanne; Valenta, Rudolf

    2013-09-01

    The major turnip (Brassica rapa) pollen allergen, belongs to a family of calcium-binding proteins (i.e., two EF-hand proteins), which occur as highly cross-reactive allergens in pollen of weeds, grasses and trees. In this study, the IgE binding capacity and allergenic activity of three recombinant allergen variants containing mutations in their calcium-binding sites were analyzed in sensitized patients with the aim to identify the most suitable hypoallergenic molecule for specific immunotherapy. Analysis of the wildtype allergen and the mutants regarding IgE reactivity and activation of basophils in allergic patients indicated that the allergen derivative mutated in both calcium-binding domains had the lowest allergenic activity. Gel filtration and circular dichroism experiments showed that both, the wildtype and the double mutant, occurred as dimers in solution and assumed alpha-helical fold, respectively. However, both fold and thermal stability were considerably reduced in the double mutant. The use of bioinformatic tools for evaluation of the solvent accessibility and charge distribution suggested that the reduced IgE reactivity and different structural properties of the double mutant may be due to a loss of negatively charged amino acids on the surface. Interestingly, immunization of rabbits showed that only the double mutant but not the wildtype allergen induced IgG antibodies which recognized the allergen and blocked binding of allergic patients IgE. Due to the extensive structural similarity and cross-reactivity between calcium-binding pollen allergens the hypoallergenic double mutant may be useful not only for immunotherapy of turnip pollen allergy, but also for the treatment of allergies to other two EF-hand pollen allergens.

  10. [Optimal measure for cultivation of Artemisia annua with high seeds yield].

    PubMed

    Wu, Yekuan; Li, Longyun; Hu, Yingi

    2009-09-01

    The relationship of Artemisia annua seed yield with density, N, P and K fertilizer applied amount was studied, and a mathematical model involving the 4 factors affecting seed yield was established using the orthogonal rotation design of quadratic regression. The seed yield function model was established according to parameters through field tests and data treated by computer techniques. The best agronomic measures complex project was selected and developed by computer imitation. The effects on seed yield of A. annua are density > N > P > K in turn. To obtain the highest yield density should be 13 000-15 000 plants x hm(-2), Ureal 186-242 kg x hm(-2), calcium superphosphate 874-1 023 kg x hm(-2), potassium chloride 135-165 kg x hm(-2) in the experiment. Reasonable planting density and fertilizer application could improve the seed yield of A. annua. PMID:19943472

  11. High-yield synthesis and optical properties of g-C3N4

    NASA Astrophysics Data System (ADS)

    Yuan, Yanwen; Zhang, Lulu; Xing, Jun; Utama, M. Iqbal Bakti; Lu, Xin; Du, Kezhao; Li, Yongmei; Hu, Xiao; Wang, Shijie; Genç, Aziz; Dunin-Borkowski, Rafal; Arbiol, Jordi; Xiong, Qihua

    2015-07-01

    Graphitic carbon nitride (g-C3N4), a metal-free semiconductor with a band gap of 2.7 eV, has received considerable attention owing to its fascinating photocatalytic performances under visible-light. g-C3N4 exhibits high thermal and chemical stability and non-toxicity such that it has been considered as the most promising photocatalyst for environmental improvement and energy conservation. Hence, it is of great importance to obtain high-quality g-C3N4 and gain a clear understanding of its optical properties. Herein, we report a high-yield synthesis of g-C3N4 products via heating of high vacuum-sealed melamine powder in an ampoule at temperatures between 450 and 650 °C. Using transmission electron microscopy (TEM), scanning transmission electron microscopy (STEM), electron energy loss spectroscopy (EELS), thermogravimetric analysis (TGA), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS), the chemical composition and crystallization of the as-produced g-C3N4 are demonstrated. A systematic optical study of g-C3N4 is carried out with several approaches. The optical phonon behavior of g-C3N4 is revealed by infrared and Raman spectroscopy, and the emission properties of g-C3N4 are investigated using photoluminescence (PL) spectroscopy, while the photocatalytic properties are explored by the photodegradation experiment.Graphitic carbon nitride (g-C3N4), a metal-free semiconductor with a band gap of 2.7 eV, has received considerable attention owing to its fascinating photocatalytic performances under visible-light. g-C3N4 exhibits high thermal and chemical stability and non-toxicity such that it has been considered as the most promising photocatalyst for environmental improvement and energy conservation. Hence, it is of great importance to obtain high-quality g-C3N4 and gain a clear understanding of its optical properties. Herein, we report a high-yield synthesis of g-C3N4 products via heating of high vacuum-sealed melamine powder in an ampoule at

  12. High Yield Synthesis of Mesoscopic Conductive and Dispersible Carbon Nanostructures via Ultrasonication of Commercial Precursors

    SciTech Connect

    Srivastava, Vikram K; Quinlan, Ronald; Agapov, Alexander L; Kisliuk, Alexander; Bhat, Gajanan; Mays, Jimmy

    2014-01-01

    The need to produce large quantities of graphenic materials displaying excellent conductivity, thermal resistance, and tunable properties for industrial applications has spurred interest in new techniques for exfoliating graphite. In this paper, sonication-assisted exfoliation of graphitic precursors in the presence of chloroform is shown to produce chemically and structurally unique exfoliated graphitic materials in high yields. These exfoliated graphites, referred to as mesographite and mesographene, respectively, exhibit unique properties which depend on the number of layers and exfoliation conditions. Structural characterization of mesographene reveals the presence of nanoscale two-dimensional graphene layers, and threedimensional carbon nanostructures sandwiched between layers, similar to those found in ball-milled and intercalated graphites. The conductivities of mesographite and mesographene are 2700 and 2000 S/m, respectively, indicating high conductivity despite flake damage. Optical absorption measurements of mesographite sonicated in various solvents showed significant changes in dispersion characteristics, and also indicated significant changes to mesoscopic colloidal behavior. A mechanism for functionalization and formation of capped carbon nanostructures is proposed by integrating the chemical and structural characterization in relation to the various carbon structures observed by electron microscopy. Composites based on common polymers were prepared by solution processing, and changes in thermal properties indicate improved dispersion of mesographite in polar polymers.

  13. Chemical and physical properties of high-yield alkaline sulfite green liquor

    SciTech Connect

    Sell, N.J.; Norman, J.C. . Natural and Applied Sciences)

    1993-11-01

    The majority of sodium sulfite pulping liquor recovery systems are based on the reductive burning of the spent liquor, followed by acidification of the resulting smelt solution by CO[sub 2]. This study investigated a number of the physical and chemical properties of the resulting green liquor which might be relevant to the optimum design of this type of sulfite and carbonate recovery system for an alkaline sulfite high-yield process. CO[sub 2] gas does generate H[sub 2]S when bubbled through green liquor; however, a large amount of solid soon is formed. Continuing the flow leads to increased amounts of H[sub 2]S, but the ratio of H[sub 2]S to CO[sub 2] remains less than 1.0. Solutions more highly concentrated in Na[sub 2]S absorb relatively more CO[sub 2], regardless of the ratios of H[sub 2]S to CO[sub 2] in the initial gas stream. The percentage of H[sub 2]S released increases with increasing Na[sub 2]S concentration. Stripping the green liquor with inert gas, steam, or vacuum does not improve the H[sub 2]S removal efficiency. The maximum CO[sub 2] pressure can be generated by decomposing pure 6 M NaHCO[sub 3]. If the starting material is a bicarbonate/carbonate mixture, conversion is incomplete and a portion of the NaHCO[sub 3] forms a dead load.

  14. Single-step, high yield synthesis of gold nanoworms and their surface enhanced Raman scattering properties

    NASA Astrophysics Data System (ADS)

    Ahmed, Waqqar; van Ruitenbeek, Jan M.

    Rod-shaped gold nanoparticles have attracted enormous attention owing to their interesting optical properties arising from the surface plasmon resonances. Slight deviation from the rod morphology can markedly change the optical properties. For-example, worm-shaped gold nanoparticles can have more than two plasmon peaks. Furthermore, they show much higher local field enhancements as compared to their rod-shaped counterparts. We have devised a simple seedless, high-yield protocol for the synthesis of gold nanoworms (NWs). NWs were grown simply by reducing HAuCl4 with ascorbic acid in a high pH reaction medium, and in the presence of growth directional agents, cetyltrimethylammonium bromide and AgNO3. In contrast to the seed-mediated growth of gold nanorods where a seed grows into a rod, NWs grow by oriental attachment of nanoparticles. By varying different reaction parameters we were able to control the length of NWs from a few nanometers to micrometers. Furthermore, the aspect ratio can also be tuned over a wide range. Gold NWs show excellent surface enhanced Raman scattering (SERS) properties. Ultra-low concentrations of various target molecules were detected using NWs based SERS substrates.

  15. A new approach to the high-yield synthesis of nanoparticles by spark discharge

    NASA Astrophysics Data System (ADS)

    Efimov, A. A.; Lizunova, A. A.; Volkov, I. A.; Mylnikov, D. A.; Arsenov, P. V.; Ivanov, V. V.

    2016-08-01

    A new approach to the high-yield synthesis of nanoparticles in the multi-gap (up to 12) spark discharge generator operating at a repetition rate as high as 2.5 kHz is discussed. This approach allows to increase the production rate by a factor of up to 100 as compared to the conventional approach utilizing single gap generator operating in the self-breakdown mode. In the case of spark erosion of aluminum electrodes in the air atmosphere, the production rate of 300 mg/h has been achieved. From the analysis of transmission electron microscopy images it was found that primary particles of the synthesized material are nearly spherical in shape and their size is 11 nm; these primary particles are united into larger objects (∼80 nm) with fractal structure. The fraction of amorphous phase is 54% and the rest 46% is of cubic γ-Al2O3 phase with the mean crystallite size of 12 nm as determined from X-ray diffraction analysis. The specific surface area of the nanoparticles determined by BET method is equal 180 m2/g.

  16. Enzymatic Process for High-Yield Turanose Production and Its Potential Property as an Adipogenesis Regulator.

    PubMed

    Park, Min-Oh; Lee, Byung-Hoo; Lim, Eunjin; Lim, Ji Ye; Kim, Yuri; Park, Cheon-Seok; Lee, Hyeon Gyu; Kang, Hee-Kwon; Yoo, Sang-Ho

    2016-06-15

    Turanose is a sucrose isomer naturally existing in honey and a promising functional sweetener due to its low glycemic response. In this study, the extrinsic fructose effect on turanose productivity was examined in Neisseria amylosucrase reaction. Turanose was produced, by increasing the amount of extrinsic fructose as a reaction modulator, with high concentration of sucrose substrate, which resulted in 73.7% of production yield. In physiological functionality test, lipid accumulation in 3T3-L1 preadipocytes in the presence of high amounts of pure glucose was attenuated by turanose substitution in a dose-dependent manner. Turanose treatments at concentrations representing 50%, 75%, and 100% of total glucose concentration in cell media significantly reduced lipid accumulation by 18%, 35%, and 72%, respectively, as compared to controls. This result suggested that turanose had a positive role in controlling adipogenesis, and enzymatic process of turanose production has a potential to develop a functional food ingredient for controlling obesity and related chronic diseases. PMID:27253611

  17. Unexpectedly low nitrogen acquisition and absence of root architecture adaptation to nitrate supply in a Medicago truncatula highly branched root mutant

    PubMed Central

    Bourion, Virginie

    2014-01-01

    To complement N2 fixation through symbiosis, legumes can efficiently acquire soil mineral N through adapted root architecture. However, root architecture adaptation to mineral N availability has been little studied in legumes. Therefore, this study investigated the effect of nitrate availability on root architecture in Medicago truncatula and assessed the N-uptake potential of a new highly branched root mutant, TR185. The effects of varying nitrate supply on both root architecture and N uptake were characterized in the mutant and in the wild type. Surprisingly, the root architecture of the mutant was not modified by variation in nitrate supply. Moreover, despite its highly branched root architecture, TR185 had a permanently N-starved phenotype. A transcriptome analysis was performed to identify genes differentially expressed between the two genotypes. This analysis revealed differential responses related to the nitrate acquisition pathway and confirmed that N starvation occurred in TR185. Changes in amino acid content and expression of genes involved in the phenylpropanoid pathway were associated with differences in root architecture between the mutant and the wild type. PMID:24706718

  18. Improved expression of human interleukin-2 in high-cell-density fermentor cultures of Escherichia coli K-12 by a phosphotransacetylase mutant.

    PubMed Central

    Bauer, K A; Ben-Bassat, A; Dawson, M; de la Puente, V T; Neway, J O

    1990-01-01

    A fluoroacetate-resistant mutant of Escherichia coli K-12 (MM-294) accumulated less acetate in the medium during growth to high cell density in fermentor cultures and was shown to be defective in its phosphotransacetylase activity. The mutant had an improved ability to continue growing during induction of interleukin-2 (IL-2) synthesis, and in fermentor cultures it gave a higher level of specific IL-2 accumulation than its parent during expression under control of the temperature-sensitive pL promoter. In flask cultures at lower cell density, the mutant again produced less acetate than the parent, although both showed a much lower level of acetate accumulation than that seen in fermentors at high cell density. Both showed a higher specific expression level of IL-2 in flask cultures, and there was a greater difference between the mutant and its parent in the final extent of specific IL-2 accumulation in fermentor cultures compared with flask cultures. Thus, the concentration of acetate in the medium, which was much higher in fermentor cultures (greater than or equal to 300 mM after 5 h of induction) than in flask cultures (less than or equal to mM) of the parent organism, was a significant factor in limiting expression of the heterologous protein product, IL-2. The acetate kinase-phosphotransacetylase pathway was therefore a major source of acetate formation in these cultures. Blocking this pathway improved accumulation of IL-2 and did not slow growth. PMID:2187412

  19. Altered Body Weight Regulation in CK1ε Null and tau Mutant Mice on Regular Chow and High Fat Diets

    PubMed Central

    Zhou, Lili; Summa, Keith C.; Olker, Christopher; Vitaterna, Martha H.; Turek, Fred W.

    2016-01-01

    Disruption of circadian rhythms results in metabolic dysfunction. Casein kinase 1 epsilon (CK1ε) is a canonical circadian clock gene. Null and tau mutations in CK1ε show distinct effects on circadian period. To investigate the role of CK1ε in body weight regulation under both regular chow (RC) and high fat (HF) diet conditions, we examined body weight on both RC and HF diets in CK1ε−/− and CK1εtau/tau mice on a standard 24 hr light-dark (LD) cycle. Given the abnormal entrainment of CK1εtau/tau mice on a 24 hr LD cycle, a separate set of CK1εtau/tau mice were tested under both diet conditions on a 20 hr LD cycle, which more closely matches their endogenous period length. On the RC diet, both CK1ε−/− and CK1εtau/tau mutants on a 24 hr LD cycle and CK1εtau/tau mice on a 20 hr LD cycle exhibited significantly lower body weights, despite similar overall food intake and activity levels. On the HF diet, CK1εtau/tau mice on a 20 hr LD cycle were protected against the development of HF diet-induced excess weight gain. These results provide additional evidence supporting a link between circadian rhythms and energy regulation at the genetic level, particularly highlighting CK1ε involved in the integration of circadian biology and metabolic physiology. PMID:27144030

  20. Attenuation of high sucrose diet–induced insulin resistance in ABC transporter deficient white mutant of Drosophila melanogaster

    PubMed Central

    Navrotskaya, Valeriya; Oxenkrug, Gregory; Vorobyova, Lyudmila; Summergrad, Paul

    2016-01-01

    Exposure to high sugar diet (HSD) is an experimental model of insulin resistance (IR) and type 2 diabetes (T2D) in mammals and insects. In Drosophila, HSD-induced IR delays emergence of pupae from larvae and eclosion of imago from pupae. Understanding of mechanisms of IR/T2D is essential for refining T2D prevention and treatment strategies. Dysregulation of tryptophan (Trp)-kynurenine (Kyn) pathway was suggested as one of the mechanisms of IR/T2D development. Rate-limiting enzyme of Trp-Kyn pathway in Drosophila is Trp 2,3-dioxygenase (TDO), an evolutionary conserved ortholog of human TDO. We previously reported attenuation of HSD-induced IR in vermilion mutants with inactive TDO. Conversion of Trp to Kyn is regulated not only by TDO activity but by intracellular Trp transport via ATP-binding cassette (ABC) transporter encoded by white gene in Drosophila. In order to evaluate the possible impact of deficient intracellular Trp transport on the inducement of IR by HSD, we compared the effect of HSD on pre-imago development in wild type flies, Canton-Special (C-S), and C-S flies containing white gene, white (C-S). Presence of white gene attenuated (by 50%) HSD-induced delay of pupae emergence from larvae and female and male imago eclosion from pupae. Present study together with our earlier report reveals that both decreased TDO activity (due to vermilion gene mutation) or deficient Trp transport into cell without affecting TDO levels (due to white gene mutation) attenuate HSD-induced development of IR in Drosophila model of T2D. Our data provide further support for hypothesis that dysregulation of Trp-Kyn pathway is one of the pathophysiological mechanisms and potential target for early diagnosis, prevention and treatment of IR/T2D. PMID:27375855

  1. High-yield synthesis of few-layer graphene flakes through electrochemical expansion of graphite in propylene carbonate electrolyte.

    PubMed

    Wang, Junzhong; Manga, Kiran Kumar; Bao, Qiaoliang; Loh, Kian Ping

    2011-06-15

    High-yield production of few-layer graphene flakes from graphite is important for the scalable synthesis and industrial application of graphene. However, high-yield exfoliation of graphite to form graphene sheets without using any oxidation process or super-strong acid is challenging. Here we demonstrate a solution route inspired by the lithium rechargeable battery for the high-yield (>70%) exfoliation of graphite into highly conductive few-layer graphene flakes (average thickness <5 layers). A negative graphite electrode can be electrochemically charged and expanded in an electrolyte of Li salts and organic solvents under high current density and exfoliated efficiently into few-layer graphene sheets with the aid of sonication. The dispersible graphene can be ink-brushed to form highly conformal coatings of conductive films (15 ohm/square at a graphene loading of <1 mg/cm(2)) on commercial paper. PMID:21557613

  2. High-yielding aquifers in crystalline basement: insights about the role of fault zones, exemplified by Armorican Massif, France

    NASA Astrophysics Data System (ADS)

    Roques, Clément; Bour, Olivier; Aquilina, Luc; Dewandel, Benoît

    2016-08-01

    While groundwater constitutes a crucial resource in many crystalline-rock regions worldwide, well-yield conditions are highly variable and barely understood. Nevertheless, it is well known that fault zones may have the capacity to ensure sustainable yield in crystalline media, but there are only a few and disparate examples in the literature that describe high-yield conditions related to fault zones in crystalline rock basements. By investigating structural and hydraulic properties of remarkable yielding sites identified in the Armorican Massif, western France, this study discusses the main factors that may explain such exceptional hydrogeological properties. Twenty-three sites, identified through analysis of databases available for the region, are investigated. Results show that: (1) the highly transmissive fractures are related to fault zones which ensure the main water inflow in the pumped wells; (2) the probability of intersecting such transmissive fault zones does not vary significantly with depth, at least within the range investigated in this study (0-200 m); and (3) high yield is mainly controlled by the structural features of the fault zones, in particular the fault dip and the presence of a connected storage reservoir. Conceptual models that summarize the hydrological properties of high-yield groundwater resources related to fault zones in crystalline basement are shown and discussed.

  3. [Effects of combined application of nitrogen and phosphorus on diurnal variation of photosynthesis at grain-filling stage and grain yield of super high-yielding wheat].

    PubMed

    Zhao, Hai-bo; Lin, Qi; Liu, Yi-guo; Jiang, Wen; Liu, Jian-jun; Zhai, Yan-ju

    2010-10-01

    Taking super high-yielding wheat cultivar Jimai 22 as test material, a field experiment was conducted to study the effects of combined application of nitrogen (N) and phosphorus (P) on the diurnal variation of photosynthesis at grain-filling stage and the grain yield of the cultivar. In treatments CK (without N and P application) and low N/P application (225 kg N x hm(-2) and 75 kg P x hm(-2)), the diurnal variation of net photosynthetic rate (Pn) was presented as double-peak curve, and there existed obvious midday depression of photosynthesis. Under reasonable application of N/P (300 kg N x hm(-2) and 150 kg P x hm(-2), treatment N2P2), the midday depression of photosynthesis weakened or even disappeared. Stomatal and non-stomatal limitations could be the causes of the midday depression. Increasing N and P supply increased the Pn, stomatal conductance (Gs), stomatal limitation value (Ls), and transpiration rate (Tr). Fertilizer P had less effects on the photosynthesis, compared with fertilizer N. When the P supply was over 150 kg x hm(-2), the increment of Pn was alleviated and even decreased. Among the fertilization treatments, treatment N2P2 had the highest Pn, Gs, and water use efficiency, being significantly different from CK. It appeared that fertilizer N had greater regulatory effect on the diurnal variation of photosynthesis, compared with fertilizer P, while the combined application of N and P had significant co-effect on the Pn, Gs, and Tr. A combined application of 300 kg N x hm(-2) and 150 kg P x hm(-2) benefited the enhancement of Pn and grain yield.

  4. High-throughput and high-yield fabrication of uniaxially-aligned chitosan-based nanofibers by centrifugal electrospinning.

    PubMed

    Erickson, Ariane E; Edmondson, Dennis; Chang, Fei-Chien; Wood, Dave; Gong, Alex; Levengood, Sheeny Lan; Zhang, Miqin

    2015-12-10

    The inability to produce large quantities of nanofibers has been a primary obstacle in advancement and commercialization of electrospinning technologies, especially when aligned nanofibers are desired. Here, we present a high-throughput centrifugal electrospinning (HTP-CES) system capable of producing a large number of highly-aligned nanofiber samples with high-yield and tunable diameters. The versatility of the design was revealed when bead-less nanofibers were produced from copolymer chitosan/polycaprolactone (C-PCL) solutions despite variations in polymer blend composition or spinneret needle gauge. Compared to conventional electrospinning techniques, fibers spun with the HTP-CES not only exhibited superior alignment, but also better diameter uniformity. Nanofiber alignment was quantified using Fast Fourier Transform (FFT) analysis. In addition, a concave correlation between the needle diameter and resultant fiber diameter was identified. This system can be easily scaled up for industrial production of highly-aligned nanofibers with tunable diameters that can potentially meet the requirements for various engineering and biomedical applications.

  5. High-pressure homogenization of raw and pasteurized milk modifies the yield, composition, and texture of queso fresco cheese.

    PubMed

    Escobar, D; Clark, S; Ganesan, V; Repiso, L; Waller, J; Harte, F

    2011-03-01

    High-pressure homogenization (HPH) of milk was studied as an alternative processing operation in the manufacturing of queso fresco cheese. Raw and pasteurized (65°C for 30 min) milks were subjected to HPH at 0, 100, 200, and 300 MPa and then used to manufacture queso fresco. The cheeses were evaluated for yield, moisture content, titratable acidity, nitrogen content, whey protein content, yield force, yield strain, and tactile texture by instrumental or trained panel analyses. The combination of HPH and thermal processing of milk resulted in cheeses with increased yield and moisture content. The net amount of protein transferred to the cheese per kilogram of milk remained constant for all treatments except raw milk processed at 300 MPa. The highest cheese yield, moisture content, and crumbliness were obtained for thermally processed milk subjected to HPH at 300 MPa. The principal component analysis of all measured variables showed that the variables yield, moisture content, and crumbliness were strongly correlated to each other and negatively correlated to the variables yield strain, protein content (wet basis), and sensory cohesiveness. It is suggested that the combination of thermal processing and HPH promotes thermally induced denaturation of whey protein, together with homogenization-induced dissociation of casein micelles. The combined effect results in queso fresco containing a thin casein-whey matrix that is able to better retain sweet whey. These results indicate that HPH has a strong potential for the manufacture of queso fresco with excellent yield and textural properties.

  6. E151 (sym15), a pleiotropic mutant of pea (Pisum sativum L.), displays low nodule number, enhanced mycorrhizae, delayed lateral root emergence, and high root cytokinin levels.

    PubMed

    Jones, James M C; Clairmont, Lindsey; Macdonald, Emily S; Weiner, Catherine A; Emery, R J Neil; Guinel, Frédérique C

    2015-07-01

    In legumes, the formation of rhizobial and mycorrhizal root symbioses is a highly regulated process which requires close communication between plant and microorganism. Plant mutants that have difficulties establishing symbioses are valuable tools for unravelling the mechanisms by which these symbioses are formed and regulated. Here E151, a mutant of Pisum sativum cv. Sparkle, was examined to characterize its root growth and symbiotic defects. The symbioses in terms of colonization intensity, functionality of micro-symbionts, and organ dominance were compared between the mutant and wild type. The endogenous cytokinin (CK) and abscisic acid (ABA) levels and the effect of the exogenous application of these two hormones were determined. E151 was found to be a low and delayed nodulator, exhibiting defects in both the epidermal and cortical programmes though a few mature and functional nodules develop. Mycorrhizal colonization of E151 was intensified, although the fungal functionality was impaired. Furthermore, E151 displayed an altered lateral root (LR) phenotype compared with that of the wild type whereby LR emergence is initially delayed but eventually overcome. No differences in ABA levels were found between the mutant and the wild type, but non-inoculated E151 exhibited significantly high CK levels. It is hypothesized that CK plays an essential role in differentially mediating the entry of the two micro-symbionts into the cortex; whereas it would inhibit the entry of the rhizobia in that tissue, it would promote that of the fungus. E151 is a developmental mutant which may prove to be a useful tool in further understanding the role of hormones in the regulation of beneficial root symbioses.

  7. E151 (sym15), a pleiotropic mutant of pea (Pisum sativum L.), displays low nodule number, enhanced mycorrhizae, delayed lateral root emergence, and high root cytokinin levels

    PubMed Central

    Jones, James M. C.; Clairmont, Lindsey; Macdonald, Emily S.; Weiner, Catherine A.; Emery, R. J. Neil; Guinel, Frédérique C.

    2015-01-01

    In legumes, the formation of rhizobial and mycorrhizal root symbioses is a highly regulated process which requires close communication between plant and microorganism. Plant mutants that have difficulties establishing symbioses are valuable tools for unravelling the mechanisms by which these symbioses are formed and regulated. Here E151, a mutant of Pisum sativum cv. Sparkle, was examined to characterize its root growth and symbiotic defects. The symbioses in terms of colonization intensity, functionality of micro-symbionts, and organ dominance were compared between the mutant and wild type. The endogenous cytokinin (CK) and abscisic acid (ABA) levels and the effect of the exogenous application of these two hormones were determined. E151 was found to be a low and delayed nodulator, exhibiting defects in both the epidermal and cortical programmes though a few mature and functional nodules develop. Mycorrhizal colonization of E151 was intensified, although the fungal functionality was impaired. Furthermore, E151 displayed an altered lateral root (LR) phenotype compared with that of the wild type whereby LR emergence is initially delayed but eventually overcome. No differences in ABA levels were found between the mutant and the wild type, but non-inoculated E151 exhibited significantly high CK levels. It is hypothesized that CK plays an essential role in differentially mediating the entry of the two micro-symbionts into the cortex; whereas it would inhibit the entry of the rhizobia in that tissue, it would promote that of the fungus. E151 is a developmental mutant which may prove to be a useful tool in further understanding the role of hormones in the regulation of beneficial root symbioses. PMID:25948707

  8. Increased episomal replication accounts for the high rate of adaptive mutation in recD mutants of Escherichia coli.

    PubMed Central

    Foster, P L; Rosche, W A

    1999-01-01

    Adaptive mutation has been studied extensively in FC40, a strain of Escherichia coli that cannot metabolize lactose (Lac-) because of a frameshift mutation affecting the lacZ gene on its episome. recD mutants of FC40, in which the exonuclease activity of RecBCD (ExoV) is abolished but its helicase activity is retained, have an increased rate of adaptive mutation. The results presented here show that, in several respects, adaptive mutation to Lac+ involves different mechanisms in recD mutant cells than in wild-type cells. About half of the apparent increase in the adaptive mutation rate of recD mutant cells is due to a RecA-dependent increase in episomal copy number and to growth of the Lac- cells on the lactose plates. The remaining increase appears to be due to continued replication of the episome, with the extra copies being degraded or passed to recD+ recipients. In addition, the increase in adaptive mutation rate in recD mutant cells is (i) dependent on activities of the single-stranded exonucleases, RecJ and ExoI, which are not required for (in fact, slightly inhibit) adaptive mutation in wild-type cells, and (ii) enhanced by RecG, which opposes adaptive mutation in wild-type cells. PMID:10224241

  9. A novel high yield method for dry functionalization of carbon nanotubes.

    PubMed

    Ansari, S G

    2011-04-01

    A novel and high yield (> 80%) dry method to functionalize (dry functionalization) carbon nanotubes (CNTs) using hydrothermal method, is reported here. The hydrothermal solution was prepared with HNO3, H2SO4 and H2O2 (1:3:2 vol. ratios) and reaction was carried out from 120 to 200 degrees C for 24 h. CNTs (multi wall) were kept in a way to avoid the direct contact with the solution. Treatment above 180 degrees C resulted in better functionalization of nanotubes as observed from Fourier transform infrared absorption spectroscopic (FTIR) measurements. Field emission scanning electron microscopic (FESEM) images showed that after functionalization, the nanotubes are seen with open ends, granular surface, twisted and are joined together. These clearly indicate the destruction of the graphite structure on the surface. This indicates that after treatment, CNTs reactivity has increased at the ends as well as at the side walls. X-ray Photoelectron Spectroscopic (XPS) studies show a shift in the C 1s peak position, increase in O 1s peak intensity and appearance of an N 1s peak. PMID:21776687

  10. High Yield Seedless Synthesis of Uniform Silver Nanoparticles with Different Sizes.

    PubMed

    Xi, Chunxiao; Song, Yahui; Zhang, Jihui; Zhang, Dongmei; Xia, Haibing

    2016-06-01

    Currently, seed-mediated growth method is the dominant strategy for synthesis of silver nanoparticles (Ag NPs) with different sizes. However, it is complicated for the researchers with limited chemistry-training. In this work, monodisperse, quasi-spherical Ag NPs with different sizes were successfully produced in high yield by adding a trace amount of NaBH4 solution into the mixtures of AgNO3, ascorbic acid and cetyltrimethylammonium chloride (CTAC) as surfactants. It was found that the sizes of as-prepared Ag NPs can be tuned from 20 nm to 50 nm by adjusting the NaBH4 concentration at optimal pH (about 6.33) and CTAC concentration (0.02-0.06 M). The surface-enhanced Raman scattering (SERS) activity of 50 nm Ag NPs shows an enhanced enhancement factor for 4-aminothiophenol (4-ATP) molecules (2.5 x 10(7) for a(1) mode). Moreover, Raman signals (a(g) mode) of 4,4'-dimercaptoazobenzene (DMAB) molecules were also observed due to the laser-induced transformation of 4-ATP molecules on the surfaces of 50 nm Ag NPs during SERS measurement. Furthermore, the transformation of probe molecules on the surfaces of metal NPs during SERS measurement have to be considered. PMID:27427638

  11. Generation of high-yield insulin producing cells from human bone marrow mesenchymal stem cells.

    PubMed

    Jafarian, Arefeh; Taghikhani, Mohammad; Abroun, Saeid; Pourpak, Zahra; Allahverdi, Amir; Soleimani, Masoud

    2014-07-01

    Allogenic islet transplantation is a most efficient approach for treatment of diabetes mellitus. However, the scarcity of islets and long term need for an immunosuppressant limits its application. Recently, cell replacement therapies that generate of unlimited sources of β cells have been developed to overcome these limitations. In this study we have described a stage specific differentiation protocol for the generation of insulin producing islet-like clusters from human bone marrow mesenchymal stem cells (hBM-MSCs). This specific stepwise protocol induced differentiation of hMSCs into definitive endoderm, pancreatic endoderm and pancreatic endocrine cells that expressed of sox17, foxa2, pdx1, ngn3, nkx2.2, insulin, glucagon, somatostatin, pancreatic polypeptide, and glut2 transcripts respectively. In addition, immunocytochemical analysis confirmed protein expression of the above mentioned genes. Western blot analysis discriminated insulin from proinsulin in the final differentiated cells. In derived insulin producing cells (IPCs), secreted insulin and C-peptide was in a glucose dependent manner. We have developed a protocol that generates effective high-yield human IPCs from hBM-MSCs in vitro. These finding suggest that functional IPCs generated by this procedure can be used as a cell-based approach for insulin dependent diabetes mellitus.

  12. High yield production of myristoylated Arf6 small GTPase by recombinant N-myristoyl transferase

    PubMed Central

    Padovani, Dominique; Zeghouf, Mahel; Traverso, José A.; Giglione, Carmela; Cherfils, Jacqueline

    2013-01-01

    Small GTP-binding proteins of the Arf family (Arf GTPases) interact with multiple cellular partners and with membranes to regulate intracellular traffic and organelle structure. Understanding the underlying molecular mechanisms requires in vitro biochemical assays to test for regulations and functions. Such assays should use proteins in their cellular form, which carry a myristoyl lipid attached in N-terminus. N-myristoylation of recombinant Arf GTPases can be achieved by co-expression in E. coli with a eukaryotic N-myristoyl transferase. However, purifying myristoylated Arf GTPases is difficult and has a poor overall yield. Here we show that human Arf6 can be N-myristoylated in vitro by recombinant N-myristoyl transferases from different eukaryotic species. The catalytic efficiency depended strongly on the guanine nucleotide state and was highest for Arf6-GTP. Large-scale production of highly pure N-myristoylated Arf6 could be achieved, which was fully functional for liposome-binding and EFA6-stimulated nucleotide exchange assays. This establishes in vitro myristoylation as a novel and simple method that could be used to produce other myristoylated Arf and Arf-like GTPases for biochemical assays. PMID:23319116

  13. High quantum yield graphene quantum dots decorated TiO2 nanotubes for enhancing photocatalytic activity

    NASA Astrophysics Data System (ADS)

    Qu, Ailan; Xie, Haolong; Xu, Xinmei; Zhang, Yangyu; Wen, Shengwu; Cui, Yifan

    2016-07-01

    Graphene quantum dots (GQDs) with high quantum yield (about 23.6% at an excitation wavelength of 320 nm) and GQDs/TiO2 nanotubes (GQDs/TiO2 NTs) composites were achieved by a simple hydrothermal method at low temperature. Photoluminescence characterization showed that the GQDs exhibited the down-conversion PL features at excitation from 300 to 420 nm and up-conversion photoluminescence in the range of 600-800 nm. The photocatalytic activity of prepared GQDs/TiO2 NTs composites on the degradation of methyl orange (MO) was significantly enhanced compared with that of pure TiO2 nanotubes (TiO2 NTs). For the composites coupling with 1.5%, 2.5% and 3.5% GQDs, the degradation of MO after 20 min irradiation under UV-vis light irradiation (λ = 380-780 nm) were 80.52%, 94.64% and 51.91%, respectively, which are much higher than that of pure TiO2 NTs (35.41%). It was inferred from the results of characterization that the improved photocatalytic activity of the GQDs/TiO2 NTs composites was attributed to the synergetic effect of up-conversion properties of the GQDs, enhanced visible light absorption and efficient separation of photogenerated electron-holes of the GQDs/TiO2 composite.

  14. High-yield room temperature route to copper sulfide hollow nanospheres and their electrochemical properties

    NASA Astrophysics Data System (ADS)

    Wang, Yiying; Li, Qing; Nie, Ming; Li, Xuelian; Li, Yuan; Zhong, Xiaolin

    2011-07-01

    CuS hollow nanospheres have been successfully synthesized in high yield by reacting anhydrous cupric sulfate (CuSO4·5H2O) with thioacetamide (TAA) in ethylene glycol (EG) with the assistance of cetyltrimethylammonium bromide (CTAB). The products were characterized systematically by XRD, EDX, FESEM, TEM and BET measurement and size analysis, CV, LSV and CP. FESEM and TEM images revealed that the as-prepared CuS hollow nanospheres had a mean diameter of about 500 nm with a hollow cavity of about 340 nm and shell thickness of about 80 nm. The spheres were constructed by numerous nanoflakes. The Brunauer-Emmett-Teller (BET) surface area of the as-synthesized products was measured to be 99.77 m2g - 1. The Barrett-Joyner-Halenda (BJH) model analysis showed that the as-prepared CuS materials had a main pore size distribution of around 25 nm. CV curves, LSV of CuS for oxygen electroreduction and CP curves showed that the as-prepared CuS nanospheres were potential candidates which can be used as cathode catalysts for the oxygen reduction reaction (ORR) in alkaline media.

  15. Tailoring wet explosion process parameters for the pretreatment of cocksfoot grass for high sugar yields.

    PubMed

    Njoku, S I; Ahring, B K; Uellendahl, H

    2013-08-01

    The pretreatment of lignocellulosic biomass is crucial for efficient subsequent enzymatic hydrolysis and ethanol fermentation. In this study, wet explosion (WEx) pretreatment was applied to cocksfoot grass and pretreatment conditions were tailored for maximizing the sugar yields using response surface methodology. The WEx process parameters studied were temperature (160-210 °C), retention time (5-20 min), and dilute sulfuric acid concentration (0.2-0.5 %). The pretreatment parameter set E, applying 210 °C for 5 min and 0.5 % dilute sulfuric acid, was found most suitable for achieving a high glucose release with low formation of by-products. Under these conditions, the cellulose and hemicellulose sugar recovery was 94 % and 70 %, respectively. The efficiency of the enzymatic hydrolysis of cellulose under these conditions was 91 %. On the other hand, the release of pentose sugars was higher when applying less severe pretreatment conditions C (160 °C, 5 min, 0.2 % dilute sulfuric acid). Therefore, the choice of the most suitable pretreatment conditions is depending on the main target product, i.e., hexose or pentose sugars.

  16. Fission Yield Measurements from Highly Enriched Uranium Irradiated Inside a Boron Carbide Capsule

    SciTech Connect

    Metz, Lori A.; Friese, Judah I.; Finn, Erin C.; Greenwood, Lawrence R.; Kephart, Rosara F.; Hines, Corey C.; King, Matthew D.; Henry, Kelley; Wall, Donald E.

    2013-05-01

    A boron carbide capsule was previously designed and tested by Pacific Northwest National Laboratory (PNNL) and Washington State University (WSU) for spectral-tailoring in mixed spectrum reactors. The presented work used this B4C capsule to create a fission product sample from the irradiation of highly enriched uranium (HEU) with a fast fission neutron spectrum. An HEU foil was irradiated inside of the capsule in WSU’s 1 MW TRIGA reactor at full power for 200 min to produce 5.8 × 1013 fissions. After three days of cooling, the sample was shipped to PNNL for radiochemical separations and analysis by gamma and beta spectroscopy. Fission yields for products were calculated from the radiometric measurements and compared to measurements from thermal neutron induced fission (analyzed in parallel with the non-thermal sample at PNNL) and published evaluated fast-pooled and thermal nuclear data. Reactor dosimetry measurements were also completed to fully characterize the neutron spectrum and total fluence of the irradiation.

  17. Isolation of a novel mutant strain of Saccharomyces cerevisiae by an ethyl methane sulfonate-induced mutagenesis approach as a high producer of bioethanol.

    PubMed

    Mobini-Dehkordi, Mohsen; Nahvi, Iraj; Zarkesh-Esfahani, Hamid; Ghaedi, Kamran; Tavassoli, Manoochehr; Akada, Rinji

    2008-04-01

    In order to obtain mutant strains showing higher bioethanol production than wild-type strains, a commercial Saccharomyces cerevisiae type was subjected to mutagenesis using ethyl methane sulfonate (EMS). After adding EMS to a shaken yeast suspension, the viability of yeast cells was assessed by diluted sample inoculation to solid yeast-extract peptone glucose (YEPG) medium at 15-min intervals. At 45 min, the viability of yeast cells was estimated to be about 40%. Mutagenized cells were recovered from YEPG broth after incubation at 30 degrees C for 18 h. After this period, EMS-treated yeast cells were grown on solid aerobic low-peptone (ALP) medium containing 2-12% (v/v) ethanol. All plates were incubated at 30 degrees C for 2-6 d in order to form colonies. The mutant strains that tolerated high concentrations of ethanol were selected for bioethanol production in microfuge tubes containing fermentation medium. Formation of bioethanol in small tubes was detected by the distillation-colorimetric method. In addition, trehalose content and invertase activity were determined in each mutant strain. Among many isolated mutant strains, there were six isolated colonies that grew on ALP medium supplemented with 10% (v/v) ethanol and one of them produced bioethanol 17.3% more than the wild type.

  18. MULTI-KEV X-RAY YIELDS FROM HIGH-Z GAS TARGETS FIELDED AT OMEGA

    SciTech Connect

    Kane, J O; Fournier, K B; May, M J; Colvin, J D; Thomas, C A; Marrs, R E; Compton, S M; Moody, J D; Bond, E J; Davis, J F

    2010-11-04

    The authors report on modeling of x-ray yield from gas-filled targets shot at the OMEGA laser facility. The OMEGA targets were 1.8 mm long, 1.95 mm in diameter Be cans filled with either a 50:50 Ar:Xe mixture, pure Ar, pure Kr or pure Xe at {approx} 1 atm. The OMEGA experiments heated the gas with 20 kJ of 3{omega} ({approx} 350 nm) laser energy delivered in a 1 ns square pulse. the emitted x-ray flux was monitored with the x-ray diode based DANTE instruments in the sub-keV range. Two-dimensional x-ray images (for energies 3-5 keV) of the targets were recorded with gated x-ray detectors. The x-ray spectra were recorded with the HENWAY crystal spectrometer at OMEGA. Predictions are 2D r-z cylindrical with DCA NLTE atomic physics. Models generally: (1) underpredict the Xe L-shell yields; (2) overpredict the Ar K-shell yields; (3) correctly predict the Xe thermal yields; and (4) greatly underpredict the Ar thermal yields. However, there are spreads within the data, e.g. the DMX Ar K-shell yields are correctly predicted. The predicted thermal yields show strong angular dependence.

  19. Biochemical characterization of temperature-induced changes in lipid metabolism in a high oleic acid mutant of Brassica rapa.

    PubMed

    Lee, M S; Guerra, D J

    1994-11-15

    We have characterized a Brassica rapa mutant (WR586) that has low levels of polyunsaturated fatty acids (D.L. Auld et al., 1992, Crop Sci. 32, 657-662). The mutant lacked oleoyl-phosphatidylcholine desaturase (ODS) activity when assayed in 6-day-old seedlings. To further characterize the mutant, the leaf fatty acid composition and major galactolipids and phospholipids were characterized in mutant (WR586) and control (cultivar "Tobin") plants grown at either 26 degrees C/26 degrees C or 10 degrees C/5 degrees C. Fatty acid profiles show significantly higher 18:1 levels in WR586 throughout 12 days of germination. The amount of saturated fatty acids decreased with a concomitant increase of 18:1. Ratios of 18:1/18:2 revealed that WR586 maintains higher mole percent of 18:1 than Tobin at all times and temperature regimes because of a lack of desaturation to 18:2. Values for monogalactosyl-diacylglycerol and digalactosyldiacylglycerol 18:1/18:2 ratios indicate a disparity in the concentration of 18:1 between WR586 and Tobin grown in either temperature during early germination. The phosphatidylcholine and phosphatidylethanolamine 18:1/18:2 ratios were higher in WR586 compared to Tobin and remained higher throughout the 12-day period. In WR586, 18:1 always accumulated to higher levels in the cooler temperature. In Tobin, 18:1 concentrations paralleled 18:2 in both temperature regimes. These results indicate that the lesion in the mutant WR586 resides at the ODS locus, 18:1 synthesis is chilling induced, and the microsomal desaturation pathway is the most prominent in early developing Brassica seedlings.

  20. High-yield production of aromatic peroxygenase by the agaric fungus Marasmius rotula

    PubMed Central

    2011-01-01

    An extracellular peroxygenase from Marasmius rotula was produced in liquid culture, chromatographically purified and partially characterized. This is the third aromatic peroxygenase (APO) that has been characterized in detail and the first one that can be produced in high yields. The highest enzyme levels of about 41,000 U l-1 (corresponding to appr. 445 mg l-1 APO protein) exceeded the hitherto reported levels more than 40-fold and were detected in carbon- and nitrogen-rich complex media. The enzyme was purified by FPLC to apparent homogeneity (SDS-PAGE) with a molecular mass of 32 kDa (27 kDa after deglycosylation) and isoelectric points between 4.97 and 5.27. The UV-visible spectrum of the native enzyme showed a characteristic maximum (Soret band) at 418 nm that shifted after reduction with sodium dithionite and flushing with carbon monoxide to 443 nm. The pH optimum of the M. rotula enzyme was found to vary between pH 5 and 6 for most reactions studied. The apparent Km-values for 2,6-dimethoxyphenol, benzyl alcohol, veratryl alcohol, naphthalene and H2O2 were 0.133, 0.118, 0.279, 0.791 and 3.14 mM, respectively. M. rotula APO was found to be highly stable in a pH range from 5 to 10 as well as in the presence of organic solvents (50% vol/vol) such as methanol, acetonitrile and N,N-dimethylformamide. Unlike other APOs, the peroxygenase of M. rotula showed neither brominating nor chlorinating activities. PMID:21988939

  1. A Low-cost, High-yield Process for the Direct Productin of High Energy Density Liquid Fuel from Biomass

    SciTech Connect

    Agrawal, Rakesh

    2014-02-21

    The primary objective and outcome of this project was the development and validation of a novel, low-cost, high-pressure fast-hydropyrolysis/hydrodeoxygenation (HDO) process (H{sub 2}Bioil) using supplementary hydrogen (H{sub 2}) to produce liquid hydrocarbons from biomass. The research efforts under the various tasks of the project have culminated in the first experimental demonstration of the H2Bioil process, producing 100% deoxygenated >C4+ hydrocarbons containing 36-40% of the carbon in the feed of pyrolysis products from biomass. The demonstrated H{sub 2}Bioil process technology (i.e. reactor, catalyst, and downstream product recovery) is scalable to a commercial level and is estimated to be economically competitive for the cases when supplementary H{sub 2} is sourced from coal, natural gas, or nuclear. Additionally, energy systems modeling has revealed several process integration options based on the H{sub 2}Bioil process for energy and carbon efficient liquid fuel production. All project tasks and milestones were completed or exceeded. Novel, commercially-scalable, high-pressure reactors for both fast-hydropyrolysis and hydrodeoxygenation were constructed, completing Task A. These reactors were capable of operation under a wide-range of conditions; enabling process studies that lead to identification of optimum process conditions. Model compounds representing biomass pyrolysis products were studied, completing Task B. These studies were critical in identifying and developing HDO catalysts to target specific oxygen functional groups. These process and model compound catalyst studies enabled identification of catalysts that achieved 100% deoxygenation of the real biomass feedstock, sorghum, to form hydrocarbons in high yields as part of Task C. The work completed during this grant has identified and validated the novel and commercially scalable H2Bioil process for production of hydrocarbon fuels from biomass. Studies on model compounds as well as real biomass

  2. The Sediment Yield of High Mountain Environment Watersheds: Strong Evidence of Transport Capacity Limitation

    NASA Astrophysics Data System (ADS)

    Micheletti, N.; Lane, S. N.; Lambiel, C.

    2015-12-01

    Alpine landscapes are likely to be particularly sensitive to climate change, because of: (1) the vulnerability of permafrost and glacial and nival processes to changes in atmospheric temperature and precipitation; (2) their history, which may have led to high rates of sediment production, and hence high rates of sediment supply; and (3) the steep slopes that may sustain sediment transport. Assessing the effects of climate change upon these processes over the timescale of recent rapid warming (i.e. decades) is difficult because of the lack of available data. Here, we use two unique data sources to assess the relative importance of these three processes for two high mountain basins (altitude range 2'000 to 3'500 m asl): (1) histories of surface change from the late 1950s, based upon construction of digital elevation models from archival imagery; and (2) a long term record, beginning in the early 1960s, of sediment export based upon the flushing of sediment from hydropower intakes. The two basins contain glaciers, rock glaciers, permafrost and the legacy of past glacial activity. These data show that the level of geomorphic activity within both basins is orders of magnitude higher than the volume of sediment exported from the basins. Decadal scale changes in elevation and surface displacement velocities suggest an acceleration of landscape dynamics from the 1980s. Changes in atmospheric temperature remain a key control of this process, but precipitation and snow cover prove critical in controlling rock glacier behaviour and can induce acceleration of surface displacements even under cold atmospheric conditions. However, the analysis of the sediment export data show that despite generally greater within basin dynamics, the rate of export of sediment remains orders of magnitude lower than rates of sediment production. Calculations of sediment transport capacity suggest that this is because the fundamental limit upon sediment yield is the capacity of the stream to

  3. [Canopy light distribution and its correlation with photosynthetic production in super-high yielding cotton fields of Xinjiang, Northwest China].

    PubMed

    Feng, Guo-Yi; Yao, Yan-Di; Luo, Hong-Hai; Zhang, Ya-Li; Du, Ming-Wei; Zhang, Wang-Feng; Xia, Dong-Li; Dong, Heng-Yi

    2012-05-01

    Taking the super-high yielding cotton fields (lint yield > or = 4000 kg x hm(-2)) in Xinjiang as the objects, this paper studied the canopy light distribution, photosynthetic rate, and dry matter accumulation at different growth stages, as well as the relationships between the characteristics of canopy light environment and the photosynthetic production. From full flowering stage to late full bolling stage, the light absorption proportion in the upper, middle and lower canopy layers in the super-high yielding cotton fields was 2:2:1, and the canopy transmission coefficients for radiation penetration and diffuse penetration were 0.20-0.55 and 0.22-0.56, respectively, being at reasonable level. The leaves in the middle and lower canopy layers could well accept light, and the leaf photosynthetic rate had little difference among different canopy layers. Compared with high yielding (3500 kg x hm(-2)) and generally high yielding (3000 kg x hm(-2)) cotton fields, super-high yielding cotton field had higher leaf area index and the highest canopy photosynthesis rate at early full boiling stage, and slowly decreased leaf area index, higher canopy photosynthesis rate, increased contribution of non-foliar organs to photosynthetic production, and larger dry matter accumulation from early boll-opening stage to full boll-opening stage. In cotton cultivation, to adjust the canopy structure for the equidistribution of light and canopy photosynthesis capacity in vertical direction could be the important strategy for the efficient utilization of absorbed light energy and the realization of super-high yielding.

  4. Enhancement of Biogas Yield of Poplar Leaf by High-Solid Codigestion with Swine Manure.

    PubMed

    Wangliang, Li; Zhikai, Zhang; Guangwen, Xu

    2016-05-01

    The aim of this work was to examine the improvement of anaerobic biodegradability of organic fractions of poplar leaf from codigestion with swine manure (SM), thus biogas yield and energy recovery. When poplar leaf was used as a sole substrate, the cumulative biogas yield was low, about 163 mL (g volatile solid (VS))(-1) after 45 days of digestion with a substrate/inoculum ratio of 2.5 and a total solid (TS) of 22 %. Under the same condition, the cumulative biogas yield of poplar leaf reached 321 mL (g VS)(-1) when SM/poplar leaf ratio was 2:5 (based on VS). The SM/poplar leaf ratio can determine C/N ratio of the cosubstrate and thus has significant influence on biogas yield. When the SM/poplar leaf ratio was 2:5, C/N ratio was calculated to be 27.02, and the biogas yield in 45 days of digestion was the highest. The semi-continuous digestion of poplar leaf was carried out with the organic loading rate of 1.25 and 1.88 g VS day(-1). The average daily biogas yield was 230.2 mL (g VS)(-1) and 208.4 mL (g VS)(-1). The composition analysis revealed that cellulose and hemicellulose contributed to the biogas production. PMID:26810922

  5. Analysis of a large dataset of mycorrhiza inoculation field trials on potato shows highly significant increases in yield.

    PubMed

    Hijri, Mohamed

    2016-04-01

    An increasing human population requires more food production in nutrient-efficient systems in order to simultaneously meet global food needs while reducing the environmental footprint of agriculture. Arbuscular mycorrhizal fungi (AMF) have the potential to enhance crop yield, but their efficiency has yet to be demonstrated in large-scale crop production systems. This study reports an analysis of a dataset consisting of 231 field trials in which the same AMF inoculant (Rhizophagus irregularis DAOM 197198) was applied to potato over a 4-year period in North America and Europe under authentic field conditions. The inoculation was performed using a liquid suspension of AMF spores that was sprayed onto potato seed pieces, yielding a calculated 71 spores per seed piece. Statistical analysis showed a highly significant increase in marketable potato yield (ANOVA, P < 0.0001) for inoculated fields (42.2 tons/ha) compared with non-inoculated controls (38.3 tons/ha), irrespective of trial year. The average yield increase was 3.9 tons/ha, representing 9.5 % of total crop yield. Inoculation was profitable with a 0.67-tons/ha increase in yield, a threshold reached in almost 79 % of all trials. This finding clearly demonstrates the benefits of mycorrhizal-based inoculation on crop yield, using potato as a case study. Further improvements of these beneficial inoculants will help compensate for crop production deficits, both now and in the future.

  6. Photosynthesis enhanced oxidative stress tolerance in high-yield rice varieties (Oryza sativa var. japonica L.) in the field.

    PubMed

    Wei, X D

    2016-01-01

    The objective of this study was to understand varietal differences in photosynthetic characteristics, chlorophyll fluorescence, antioxidant capability, and yield of japonica rice varieties. Nanjing 44, Oryza sativa var. japonica (average yield of 12.7 t/ha), Nanjing 46, and Nanjing 5055 (average yields of 11.3 and 11.5 t/ha) were included as "super" and high-yield varieties, respectively, whereas Wuyunjing 7 (average yield of 10.2 t/ha) was included as a control variety. These varieties were grown under field conditions in Jiangsu Province, China, in 2010-2012. Different organs (panicle, grain, etc.) were measured, before and after flowering, to identify differences of dry matter accumulation and transformation properties. Photosynthesis, the chlorophyll content, and antioxidant enzyme activities of the flag leaf in the days after flowering (DAF) were also investigated. The results showed that, compared with the other three rice varieties, Nanjing 44 had the highest plant dry weight and number of grains per panicle. It also had a relatively high net flag leaf photosynthetic rate and showed the least inhibition of photosynthesis at noon in DAF, which probably explains the higher yield in this variety. Furthermore, Nanjing 44 also had the highest stem export and conversion rate from stem to grain, exhibiting a strong ability to convert and distribute photosynthetic products. After DAF 42, Nanjing 44 still maintained a high-soluble protein content and a high antioxidant ability in the leaves to clear peroxidation products, which could protect the photosynthetic apparatus of the flag leaves, and maintain the grain-filling activity for longer. The high-yield capability of Nanjing 44 was attributed to its photosynthetic advantages in the leaves during the late developmental stage. PMID:27525885

  7. Photoyellowing inhibition of bleached high yield pulps using novel water-soluble UV screens.

    PubMed

    Argyropoulos, D S; Halevy, P; Peng, P

    2000-02-01

    To address the deficiencies of benzophenone UV screens for preventing brightness reversion in high yield mechanical papers, we synthesized a new series of such materials with enhanced water solubility and compatibility with the lignocellulosic substrate. A series of 2,4-dihydroxybenzophenones (DHB) were synthesized containing various Mannich bases at the C3 position of one of its rings. They possess the UV-screening ability of o-hydroxylbenzophenones, and they also contain tertiary nitrogen atoms that may function as radical scavengers. Aqueous solutions of the hydrochloride salt of 3-(dimethylaminomethylene)-2,4-dihydroxylbenzophenone (1), when applied on bleached chemithermomechanical pulp (CTMP) sheets, were significantly more efficient in preventing photoyellowing than the original DHB applied on the sheets from ethanol-water solutions. This confirmed our original hypothesis that increasing the compatibility of the UV screen with the lignocellulosic matrix would increase its efficiency in preventing photoyellowing. Compound 1, however, was found to be somewhat more effective than its hydrochloride salt toward preventing photoyellowing. This was attributed to the synergistic action of the free tertiary aminic center attached on the molecule with its UV-screening ability. To comprehend further the various parameters that influence the photoyellowing inhibition performance of these compounds and DHB with bleached CTMP pulp fibers, a series of handsheets were prepared at different pH. The interactions of the protonated compound 1 with pulp fibers were then evaluated by studying their kinetics of absorption and desorption to and from the fiber matrix. This part of our study found that the adsorption of protonated Mannich derivatives of DHB onto pulp is most likely governed by a cation-exchange mechanism involving the cationic amine group with the sulfonic and carboxylic acid groups located on the surface of the fibers. The pH the paper sheet was made from was also

  8. [Characteristics of canopy structure of super high yielding japonica hybrid rice community].

    PubMed

    Chen, Jinhong; Zhang, Guoping; Guo, Hengde; Mao, Guojuan

    2003-06-01

    In this paper, the characteristics of canopy structure, such as the numbers of seedling, panicle and grain, the distribution of dry matters in different canopy layers and different organs, and the distributions of LAI and of solar radiation in different canopy layers of super high yielding community of japonica hybrid rice were studied, in comparison with normal japonica rice. The results showed that the total the dry matter weight and the dry matter weight of layers below 40 cm, 40-60 cm, 60-80 cm and above 80 cm of japonica hybrid rice canopy were 32.29%, 29.12%, 13.95%, 16.45% and 100.17% higher those that of normal japonica rice, respectively. The ratios of dry leaf (photosynthetic organ) and of dry panicle (sink organ) weight to total dry weight were 24.8% and 12.8%, respectively, which were greater than those of normal japonica rice, while the ratios of dry sheath and stem (storage organs) weight were 33.6% and 28.9%, respectively, which were lower than those of normal japonica rice. The allotment of LAI in different layers of japonica hybrid rice canopy was reasonable, and the LAI of above 40 cm layer at full heading stage reached 5.44. The solar radiation was well-distributed inside japonica hybrid rice canopy, for example, the solar radiation in layers below 60 cm were 13.1%-37.0% higher, but 5.9%-12.2% lower above 60 cm than that of normal japonica rice. The extinction coefficients of solar radiation in layers below 20 cm, 20-40 cm, 40-60 cm and 60-80 cm of japonica hybrid rice canopy were 35.1%, 13.5%, 29.1% and 17.2% lower than that of normal japonica rice, respectively.

  9. A New Strategy for Production of 5-Aminolevulinic Acid in Recombinant Corynebacterium glutamicum with High Yield

    PubMed Central

    Yang, Peng; Liu, Wenjing; Cheng, Xuelian; Wang, Jing; Qi, Qingsheng

    2016-01-01

    ABSTRACT 5-Aminolevulinic acid (ALA), a nonprotein amino acid involved in tetrapyrrole synthesis, has been widely applied in agriculture, medicine, and food production. Many engineered metabolic pathways have been constructed; however, the production yields are still low. In this study, several 5-aminolevulinic acid synthases (ALASs) from different sources were evaluated and compared with respect to their ALA production capacities in an engineered Corynebacterium glutamicum CgS1 strain that can accumulate succinyl-coenzyme A (CoA). A codon-optimized ALAS from Rhodobacter capsulatus SB1003 displayed the best potential. Recombinant strain CgS1/pEC-SB produced 7.6 g/liter ALA using a mineral salt medium in a fed-batch fermentation mode. Employing two-stage fermentation, 12.46 g/liter ALA was produced within 17 h, with a productivity of 0.73 g/liter/h, in recombinant C. glutamicum. Through overexpression of the heterologous nonspecific ALA exporter RhtA from Escherichia coli, the titer was further increased to 14.7 g/liter. This indicated that strain CgS1/pEC-SB-rhtA holds attractive industrial application potential for the future. IMPORTANCE In this study, a two-stage fermentation strategy was used for production of the value-added nonprotein amino acid 5-aminolevulinic acid from glucose and glycine in a generally recognized as safe (GRAS) host, Corynebacterium glutamicum. The ALA titer represented the highest in the literature, to our knowledge. This high production capacity, combined with the potential easy downstream processes, made the recombinant strain an attractive candidate for industrial use in the future. PMID:26921424

  10. In search of annual legumes to improve forage sorghum yield and nutritive value in the southern high plains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Livestock production is significant in the Southern High Plains of the USA and demand is increasing for greater forage dry matter (DM) yield with increased nutritive value. Forage sorghum (FS)[Sorghum bicolor (L.) Moench] is commonly used, although, it is low in crude protein (CP) and high in fiber....

  11. The Maize High-Lysine Mutant opaque7 Is Defective in an Acyl-CoA Synthetase-Like Protein

    PubMed Central

    Miclaus, Mihai; Wu, Yongrui; Xu, Jian-Hong; Dooner, Hugo K.; Messing, Joachim

    2011-01-01

    Maize (Zea mays) has a large class of seed mutants with opaque or nonvitreous endosperms that could improve the nutritional quality of our food supply. The phenotype of some of them appears to be linked to the improper formation of protein bodies (PBs) where zein storage proteins are deposited. Although a number of genes affecting endosperm vitreousness have been isolated, it has been difficult to clone opaque7 (o7), mainly because of its low penetrance in many genetic backgrounds. The o7-reference (o7-ref) mutant arose spontaneously in a W22 inbred, but is poorly expressed in other lines. We report here the isolation of o7 with a combination of map-based cloning and transposon tagging. We first identified an o7 candidate gene by map-based cloning. The putative o7-ref allele has a 12-bp in-frame deletion of codons 350–353 in a 528-codon-long acyl-CoA synthetase-like gene (ACS). We then confirmed this candidate gene by generating another mutant allele from a transposon-tagging experiment using the Activator/Dissociation (Ac/Ds) system in a W22 background. The second allele, isolated from ∼1 million gametes, presented a 2-kb Ds insertion that resembles the single Ds component of double-Ds, McClintock’s original Dissociation element, at codon 496 of the ACS gene. PBs exhibited striking membrane invaginations in the o7-ref allele and a severe number reduction in the Ds-insertion mutant, respectively. We propose a model in which the ACS enzyme plays a key role in membrane biogenesis, by taking part in protein acylation, and that altered PBs render the seed nonvitreous. PMID:21926304

  12. Control and reduction of immersion defectivity for yield enhancement at high volume production

    NASA Astrophysics Data System (ADS)

    Nakano, Katsushi; Seki, Rei; Sekito, Toshiyuki; Yoshida, Masato; Fujiwara, Tomoharu; Iriuchijima, Yasuhiro; Owa, Soichi

    2009-03-01

    Volume device manufacturing using immersion lithography is widely accepted as the solution for patterning IC features below 40 nm half pitch. In order to ensure high yield and steady productivity tight control of defectivity is essential. A major source of defects and tool contamination is the particles introduced by incoming wafers. Particles can be categorized in two groups: particles attached to wafer surface or residues on the wafer edge. Surface or edge peeling of topcoats can also be a source of particle. Adhesion force between topcoat or topcoat-less (TC-less) resist and wafer is one of the most important parameter for particle reduction. Peeling test results proved that TC-less resist has better adhesion performance than topcoat. One of the most commonly used adhesion promoting material is hexamethyldisilazane (HMDS). Application condition of this material is an important factor in preventing wafer edge and surface topcoat peeling. Studies have shown lower temperature and longer application of HMDS shows better adhesion result. Maintaining a clean wafer surface is also a very important factor for particle reduction. Pre-rinse, which can rinse off particles before exposure, was evaluated and the efficiency was confirmed. Edge particles are more effectively reduced by pre-rinse, because weakly attached topcoat and wafer edge residues were effectively removed by pre-rinse. For further particle reduction, edge residue reduction and cut line roughness improvement were evaluated and their effectiveness was confirmed. Lower cut position achieved improved particle counts on both topcoat and TC-less resist; more frequent contact between water and cut-line can weaken the adhesion and consequently peel off topcoat or TC-less resist. Finally the relationship between defectivity and hydrophobicity is analyzed, high Receding Contact Angle (RCA) showed better defectivity result. Topcoat and TC-less process is compared for each defectivity reduction methodology and for

  13. Biosolar cells: global artificial photosynthesis needs responsive matrices with quantum coherent kinetic control for high yield.

    PubMed

    Purchase, R L; de Groot, H J M

    2015-06-01

    propose that synergy gains by such processes form a basis for further progress towards high efficiency and yield for a global project on artificial photosynthesis. Finally, we look at artificial photosynthesis research in The Netherlands and use this as an example of how an interdisciplinary approach is beneficial to artificial photosynthesis research. We conclude with some of the potential societal consequences of a large-scale roll out of artificial photosynthesis. PMID:26052428

  14. Biosolar cells: global artificial photosynthesis needs responsive matrices with quantum coherent kinetic control for high yield

    PubMed Central

    Purchase, R. L.; de Groot, H. J. M.

    2015-01-01

    . We propose that synergy gains by such processes form a basis for further progress towards high efficiency and yield for a global project on artificial photosynthesis. Finally, we look at artificial photosynthesis research in The Netherlands and use this as an example of how an interdisciplinary approach is beneficial to artificial photosynthesis research. We conclude with some of the potential societal consequences of a large-scale roll out of artificial photosynthesis. PMID:26052428

  15. Biosolar cells: global artificial photosynthesis needs responsive matrices with quantum coherent kinetic control for high yield.

    PubMed

    Purchase, R L; de Groot, H J M

    2015-06-01

    propose that synergy gains by such processes form a basis for further progress towards high efficiency and yield for a global project on artificial photosynthesis. Finally, we look at artificial photosynthesis research in The Netherlands and use this as an example of how an interdisciplinary approach is beneficial to artificial photosynthesis research. We conclude with some of the potential societal consequences of a large-scale roll out of artificial photosynthesis.

  16. Cesium hafnium chloride: A high light yield, non-hygroscopic cubic crystal scintillator for gamma spectroscopy

    SciTech Connect

    Burger, Arnold; Rowe, Emmanuel; Groza, Michael; Morales Figueroa, Kristle; Cherepy, Nerine J.; Beck, Patrick R.; Hunter, Steven; Payne, Stephen A.

    2015-10-05

    We report on the scintillation properties of Cs{sub 2}HfCl{sub 6} (cesium hafnium chloride or CHC) as an example of a little-known class of non-hygroscopic compounds having the generic cubic crystal structure of K{sub 2}PtCl{sub 6}. The crystals are easily growable from the melt using the Bridgman method with minimal precursor treatments or purification. CHC scintillation is centered at 400 nm, with a principal decay time of 4.37 μs and a light yield of up to 54 000 photons/MeV when measured using a silicon CCD photodetector. The light yield is the highest ever reported for an undoped crystal, and CHC also exhibits excellent light yield nonproportionality. These desirable properties allowed us to build and test CHC gamma-ray spectrometers providing energy resolution of 3.3% at 662 keV.

  17. Energy transfer and light yield properties of a new highly loaded indium(III) β-diketonate organic scintillator system

    NASA Astrophysics Data System (ADS)

    Buck, C.; Hartmann, F. X.; Motta, D.; Schoenert, S.

    2007-02-01

    We present combined experimental and model studies of the light yield and energy transfer properties of a newly developed high light yield scintillator based on indium(III)-tris(2,4-pentanedionate) in a 2-(4-biphenyl)-5-phenyloxazole (BPO), methoxybenzene organic liquid; of interest to the detection of solar electron neutrino oscillations. Optical measurements are made to understand the energy transfer properties and a model is advanced to treat the unusual conditions of high metal and fluor loadings. Such scintillator types are of interest to the exploration of novel luminescent materials and the development of large-scale detectors for studying fundamental properties of naturally occurring neutrinos.

  18. Characterization of low-acetic-acid-producing yeast isolated from 2-deoxyglucose-resistant mutants and its application to high-gravity brewing.

    PubMed

    Mizuno, Akihiro; Tabei, Hideaki; Iwahuti, Masahumi

    2006-01-01

    We isolated a mutant with low acetic acid and high ethanol productivities from 2-deoxyglucose-resistant mutants of brewers' yeast NCYC1245 (Saccharomyces cerevisiae). To determine the mechanism for these properties in the mutant (2DGR19) during fermentation, gene expression and enzyme activity related to acetic acid and ethanol production were investigated. DNA microarray analysis revealed that the transcriptional levels of many genes involved in glycolysis were higher in 2DGR19 than in NCYC1245. Among these transcriptional levels of 2DGR19 relative to NCYC1245, the expression level of ADH4 encoding alcohol dehydrogenase (ADH) was highest, which corresponded to the high ADH activity in 2DGR19. Quantitative PCR analysis also revealed that the transcriptional level of ADH4 was the highest among ADH1 to ADH4. Although no significant differences in the transcriptional levels of ALD2 to ALD6 encoding acetaldehyde dehydrogenase (ALD) between 2DGR19 and NCYC1245 were observed, ALD activity in 2DGR19 was lower. Using quantitative PCR analysis, ALD6 was found to be the most highly expressed among the ALD2 to ALD6 genes. These results indicate that ALD6 contributes to a low ALD activity, depending on post-transcriptional regulation. A high ADH activity appeared to be the major reason for the high ethanol productivity of 2DGR19. A low ALD activity was considered to be principally responsible for a low acetic acid productivity, although a high ADH activity also might have played a role. Beer brewed using 2DGR19 in pilot-scale high-gravity brewing contained about half as much acetic acid and 1.1% more ethanol compared with that brewed using NCYC1245. The use of 2DGR19 may overcome difficulties associated with high-gravity brewing.

  19. Mapping of barley alpha-amylases and outer subsite mutants reveals dynamic high-affinity subsites and barriers in the long substrate binding cleft.

    PubMed

    Kandra, Lili; Hachem, Maher Abou; Gyémánt, Gyöngyi; Kramhøft, Birte; Svensson, Birte

    2006-09-18

    Subsite affinity maps of long substrate binding clefts in barley alpha-amylases, obtained using a series of maltooligosaccharides of degree of polymerization of 3-12, revealed unfavorable binding energies at the internal subsites -3 and -5 and at subsites -8 and +3/+4 defining these subsites as binding barriers. Barley alpha-amylase 1 mutants Y105A and T212Y at subsite -6 and +4 resulted in release or anchoring of bound substrate, thus modifying the affinities of other high-affinity subsites (-2 and +2) and barriers. The double mutant Y105A-T212Y displayed a hybrid subsite affinity profile, converting barriers to binding areas. These findings highlight the dynamic binding energy distribution and the versatility of long maltooligosaccharide derivatives in mapping extended binding clefts in alpha-amylases.

  20. High-throughput FACS-based mutant screen identifies a gain-of-function allele of the Fusarium graminearum adenylyl cyclase causing deoxynivalenol over-production.

    PubMed

    Blum, Ailisa; Benfield, Aurélie H; Stiller, Jiri; Kazan, Kemal; Batley, Jacqueline; Gardiner, Donald M

    2016-05-01

    Fusarium head blight and crown rot, caused by the fungal plant pathogen Fusarium graminearum, impose a major threat to global wheat production. During the infection, plants are contaminated with mycotoxins such as deoxynivalenol (DON), which can be toxic for humans and animals. In addition, DON is a major virulence factor during wheat infection. However, it is not fully understood how DON production is regulated in F. graminearum. In order to identify regulators of DON production, a high-throughput mutant screen using Fluorescence Activated Cell Sorting (FACS) of a mutagenised TRI5-GFP reporter strain was established and a mutant over-producing DON under repressive conditions identified. A gain-of-function mutation in the F. graminearum adenylyl cyclase (FAC1), which is a known positive regulator of DON production, was identified as the cause of this phenotype through genome sequencing and segregation analysis. Our results show that the high-throughput mutant screening procedure developed here can be applied for identification of fungal proteins involved in diverse processes. PMID:26932301

  1. Identification of differentially expressed genes in sorghum (Sorghum bicolor) brown midrib mutants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sorghum (Sorghum bicolor L.), with a high biomass yield and excellent tolerance to drought and low nutrition, has been recommended as one of the most competitive bioenergy crops. Brown midrib (bmr) mutant sorghum with reduced lignin content showed a high potential for the improvement of bioethanol ...

  2. Innovations in papermaking: an LCA of printing and writing paper from conventional and high yield pulp.

    PubMed

    Manda, B M Krishna; Blok, Kornelis; Patel, Martin K

    2012-11-15

    Pulp and paper industry is facing challenges such as resource scarcity and greenhouse gas (GHG) emissions. The objective of this research is to investigate whether the use of new coatings (micro or nano TiO(2)) and different pulp types could bring savings in wood, energy, GHG emissions and other environmental impacts in comparison with conventional printing and writing paper. We studied three types of pulp, namely i) unbleached virgin kraft pulp, ii) recovered fiber, and iii) high yield virgin chemithermo-mechanical pulp (CTMP). A life cycle assessment (LCA) was conducted from cradle to grave. Applying attributional modeling, we found that wood savings amount to 60% for the nanoparticle coated recovered fiber paper and 35% for the micro TiO(2) coated CTMP paper. According to the ReCiPe single score impact assessment method, the new product configurations allow the reduction of the environmental impacts by 10-35% compared to conventional kraft paper. Applying consequential modeling, we found larger energy and GHG emission savings compared to attributional modeling because the saved wood is used for producing energy, thereby replacing fossil fuels. The nanoparticle coated recovered fiber paper offered savings of non-renewable energy use (NREU) by 100% (13GJ/ton paper) and GHG emission reduction by 75% (0.6 tonCO(2)eq./ton paper). Micro TiO(2) coated CTMP paper offered NREU savings by 25% (3GJ/ton paper) and savings of GHG emissions by 10% (0.1 tonCO(2)eq./ton paper). The taking into account of all environmental impacts with the ReCiPe single score method leads to comparable results as that of attributional modeling. We conclude that the nanoparticle coated recovered fiber paper offered the highest savings and lowest environmental impacts. However, human toxicity and ecotoxicity impacts of the nanoparticles were not included in this analysis and need further research. If this leads to the conclusion that the toxicity impacts of the nanoparticles are serious, then the

  3. Innovations in papermaking: an LCA of printing and writing paper from conventional and high yield pulp.

    PubMed

    Manda, B M Krishna; Blok, Kornelis; Patel, Martin K

    2012-11-15

    Pulp and paper industry is facing challenges such as resource scarcity and greenhouse gas (GHG) emissions. The objective of this research is to investigate whether the use of new coatings (micro or nano TiO(2)) and different pulp types could bring savings in wood, energy, GHG emissions and other environmental impacts in comparison with conventional printing and writing paper. We studied three types of pulp, namely i) unbleached virgin kraft pulp, ii) recovered fiber, and iii) high yield virgin chemithermo-mechanical pulp (CTMP). A life cycle assessment (LCA) was conducted from cradle to grave. Applying attributional modeling, we found that wood savings amount to 60% for the nanoparticle coated recovered fiber paper and 35% for the micro TiO(2) coated CTMP paper. According to the ReCiPe single score impact assessment method, the new product configurations allow the reduction of the environmental impacts by 10-35% compared to conventional kraft paper. Applying consequential modeling, we found larger energy and GHG emission savings compared to attributional modeling because the saved wood is used for producing energy, thereby replacing fossil fuels. The nanoparticle coated recovered fiber paper offered savings of non-renewable energy use (NREU) by 100% (13GJ/ton paper) and GHG emission reduction by 75% (0.6 tonCO(2)eq./ton paper). Micro TiO(2) coated CTMP paper offered NREU savings by 25% (3GJ/ton paper) and savings of GHG emissions by 10% (0.1 tonCO(2)eq./ton paper). The taking into account of all environmental impacts with the ReCiPe single score method leads to comparable results as that of attributional modeling. We conclude that the nanoparticle coated recovered fiber paper offered the highest savings and lowest environmental impacts. However, human toxicity and ecotoxicity impacts of the nanoparticles were not included in this analysis and need further research. If this leads to the conclusion that the toxicity impacts of the nanoparticles are serious, then the

  4. New germplasm lines with high yield and fiber quality combined with nematode resistance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The lines BAR-8, -11, -13, -25, -33, -41, and -48 were developed to have both resistance to nematodes and superior yield and quality. All have resistance to reniform nematodes derived from G. barbadense GB 713 and several carry the DNA marker for Mi1, the gene for resistance to root-knot nematodes....

  5. Characteristics of a glycerol utilization mutant of Neurospora crassa.

    PubMed Central

    Courtright, J B

    1975-01-01

    A mutant of Neurospora crassa able to grow on liquid minimal glycerol medium without evidence of conidiation and with high cell yields has been isolated and shown to be allelic to ff-1. The glycerol-specific induction of glycerokinase and glycerol-3-phosphate dehydrogenase was similar in both wild-type and mutant cells, although higher specific activities as well as higher glycerokinase cross-reacting material levels were found in fully induced mutant cells. After growth in minimal glycerol medium there is a significant reduction in wild-type cells of the activities of both pyruvate dehydrogenase and dihydrolipoyl transacetylase. This evidence indicates a relationship between the conditional acetate requirement by wild-type cells grown on glycerol medium and the levels of the pyruvate dehydrogenase complex. Images PMID:126227

  6. Construction and characterization of Pseudomonas aeruginosa algB mutants: role of algB in high-level production of alginate.

    PubMed Central

    Goldberg, J B; Ohman, D E

    1987-01-01

    The algB gene, which is involved in the production of alginate in Pseudomonas aeruginosa, was localized to approximately 2.2 kilobases of DNA from strain FRD by using transposon Tn501 insertion mutagenesis, subcloning, and complementation techniques. The previously reported alg-50(Ts) mutation, which confers the phenotype of temperature-sensitive alginate production, was here designated as an algB allele. A transduction-mediated gene replacement technique was used for site-directed mutagenesis to isolate and characterize algB::Tn501 mutants of P. aeruginosa FRD. Although algB::Tn501 mutants had a nonmucoid phenotype (indicating an alginate deficiency), they still produced about 1 to 5% of wild-type levels of alginate in most growth media and up to 16% in very rich media. The algB::Tn501 mutations had no apparent effect on growth rate or growth requirements. Using another gene replacement technique called excision marker rescue, we constructed a chromosomal algB deletion (delta algB) mutant of P. aeruginosa FRD. The delta algB mutant also produced low levels of alginate as did the algB::Tn501 mutants. The alginate produced by algB::Tn501 mutants resembled wild-type alginate by all criteria studied: molecular weight, acetylation, and proportion of mannuronic and guluronic acids. Thus, the algB gene product is apparently involved in the high-level production of alginate by P. aeruginosa and is not directly involved in the pathway leading to its biosynthesis. Chromosomal mapping of an algB::Tn501 insertion showed linkage to the trp-2 marker on the FRD chromosome as does the algB50(Ts) mutation. The excision marker rescue technique was also used to place the algB::Tn501 marker on the chromosome of characterized strains of P. aeruginosa PAO. The algB::Tn501 mutation mapped near 21 min on the PAO chromosome. Images PMID:3031015

  7. High-yield membrane protein expression from E. coli using an engineered outer membrane protein F fusion.

    PubMed

    Su, Pin-Chuan; Si, William; Baker, Deidre L; Berger, Bryan W

    2013-04-01

    Obtaining high yields of membrane proteins necessary to perform detailed structural study is difficult due to poor solubility and variability in yields from heterologous expression systems. To address this issue, an Escherichia coli-based membrane protein overexpression system utilizing an engineered bacterial outer membrane protein F (pOmpF) fusion has been developed. Full-length human receptor activity-modifying protein 1 (RAMP1) was expressed using pOmpF, solubilized in FC15 and purified to homogeneity. Using circular dichroism and fluorescence spectroscopy, purified full-length RAMP1 is composed of approximately 90% α-helix, and retains its solubility and structure in FC15 over a wide range of temperatures (20-60°C). Thus, our approach provides a useful, complementary approach to achieve high-yield, full-length membrane protein overexpression for biophysical studies.

  8. High-Level Production of Exopolysaccharides by a Cosmic Radiation-Induced Mutant M270 of the Maitake Medicinal Mushroom, Grifola frondosa (Agaricomycetes).

    PubMed

    Zhao, Chen; Tian, Xue-Mei; Wang, Guang-Yuan; Song, Ai-Rong; Liang, Wen-Xing

    2016-01-01

    A new Grifola frondosa mutant, M270, was successfully isolated for high production of exopolysaccharides (EPSs) using cosmic radiation-induced mutagenesis. We found that the mutant M270 had a clearer and thicker EPS layer (~10 µm) adhering to mycelia than those of its parent strain 265 after Congo red staining. In the 20-L batch fermentation for M270, 10.3 g/L of EPS and 17.9 g/L of dry mycelia biomass were obtained after 204 hours of fermentation. Furthermore, a main water-soluble fraction (EP1) in the EPS was purified from M270 and then confirmed to be heteroglycan-protein complex with 91% (w/w) total carbohydrates and 9% (w/w) total proteins. Four kinds of monosaccharide-D-mannose, D-glucosamine, D-glucose, and D-xylose-were detected in EP1 with a molar ratio of 17.6:1.8:100:2.5. The molecular mass of the main component in EP1 was 8.9 kDa. The EPS from M270 significantly inhibited the growth of sarcoma 180 solid tumors in mice. This G. frondosa M270 mutant could serve as a better candidate strain for polysaccharide production.

  9. High-Throughput Genotyping of Green Algal Mutants Reveals Random Distribution of Mutagenic Insertion Sites and Endonucleolytic Cleavage of Transforming DNA[W][OPEN

    PubMed Central

    Zhang, Ru; Patena, Weronika; Armbruster, Ute; Gang, Spencer S.; Blum, Sean R.; Jonikas, Martin C.

    2014-01-01

    A high-throughput genetic screening platform in a single-celled photosynthetic eukaryote would be a transformative addition to the plant biology toolbox. Here, we present ChlaMmeSeq (Chlamydomonas MmeI-based insertion site Sequencing), a tool for simultaneous mapping of tens of thousands of mutagenic insertion sites in the eukaryotic unicellular green alga Chlamydomonas reinhardtii. We first validated ChlaMmeSeq by in-depth characterization of individual insertion sites. We then applied ChlaMmeSeq to a mutant pool and mapped 11,478 insertions, covering 39% of annotated protein coding genes. We observe that insertions are distributed in a manner largely indistinguishable from random, indicating that mutants in nearly all genes can be obtained efficiently. The data reveal that sequence-specific endonucleolytic activities cleave the transforming DNA and allow us to propose a simple model to explain the origin of the poorly understood exogenous sequences that sometimes surround insertion sites. ChlaMmeSeq is quantitatively reproducible, enabling its use for pooled enrichment screens and for the generation of indexed mutant libraries. Additionally, ChlaMmeSeq allows genotyping of hits from Chlamydomonas screens on an unprecedented scale, opening the door to comprehensive identification of genes with roles in photosynthesis, algal lipid metabolism, the algal carbon-concentrating mechanism, phototaxis, the biogenesis and function of cilia, and other processes for which C. reinhardtii is a leading model system. PMID:24706510

  10. High-Level Production of Exopolysaccharides by a Cosmic Radiation-Induced Mutant M270 of the Maitake Medicinal Mushroom, Grifola frondosa (Agaricomycetes).

    PubMed

    Zhao, Chen; Tian, Xue-Mei; Wang, Guang-Yuan; Song, Ai-Rong; Liang, Wen-Xing

    2016-01-01

    A new Grifola frondosa mutant, M270, was successfully isolated for high production of exopolysaccharides (EPSs) using cosmic radiation-induced mutagenesis. We found that the mutant M270 had a clearer and thicker EPS layer (~10 µm) adhering to mycelia than those of its parent strain 265 after Congo red staining. In the 20-L batch fermentation for M270, 10.3 g/L of EPS and 17.9 g/L of dry mycelia biomass were obtained after 204 hours of fermentation. Furthermore, a main water-soluble fraction (EP1) in the EPS was purified from M270 and then confirmed to be heteroglycan-protein complex with 91% (w/w) total carbohydrates and 9% (w/w) total proteins. Four kinds of monosaccharide-D-mannose, D-glucosamine, D-glucose, and D-xylose-were detected in EP1 with a molar ratio of 17.6:1.8:100:2.5. The molecular mass of the main component in EP1 was 8.9 kDa. The EPS from M270 significantly inhibited the growth of sarcoma 180 solid tumors in mice. This G. frondosa M270 mutant could serve as a better candidate strain for polysaccharide production. PMID:27649730

  11. A strategic study using mutant-strain entrapment in calcium alginate for the production of Saccharomyces cerevisiae cells with high invertase activity.

    PubMed

    Rossi-Alva, Juan Carlos; Rocha-Leão, Maria Helena Miguez

    2003-08-01

    Entrapped cells and entrapped cells grown inside of a calcium alginate matrix as well as free cells have been investigated using Saccharomyces cerevisiae mutant strains with regard to their pattern of growth and invertase activity. The repression of invertase by glucose and glucose-consumption ability were considered in the selection process of the mutants. Efficient sucrose bioconversion due to high invertase activity was obtained when entrapped mutant strain Q6R2 cells were grown within calcium alginate gel beads using sucrose plus glucose as the carbon source. Under these conditions, 1 mg (dry weight) of entrapped cells is able to produce 20 micromol of inverted sugar in 3 min (the maximum activity obtained was 20 units x mg(-1)). The experiments were carried out for 6 months without appreciable loss of either bead integrity or invertase activity. The biocatalyst was also stored at 4 degrees C for 6 months without appreciable loss of the invertase activity. This work shows that entrapped yeast cells with a weak ability to consume sugar may be used to produce inverted sugar.

  12. REGIONAL ANALYSIS OF INORGANIC NITROGEN YIELD AND RETENTION IN HIGH-ELEVATION ECOSYSTEMS OF THE SIERRA NEVADA AND ROCKY MOUNTAINS

    EPA Science Inventory

    Yields and retention of inorganic nitrogen (DIN) and nitrate concentrations in surface runoff are summarized for 28 high elevation watersheds in the Sierra Nevada, California and Rocky Mountains of Wyoming and Colorado. Catchments ranged in elevation from 2475 to 3603 m and from...

  13. Simulating evapotranspiration (ET) and corn yield response to irrigation management in the Texas High Plains using DSSAT

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Grain corn (Zea mays L) continues to be a major irrigated crop in the northern Texas High Plains. Improvements in irrigation system efficiency, irrigation management, and plant genetics have increased average yields while decreasing seasonal water use in the last 40 years. However, declining water l...

  14. Registration of ‘USDA-N8002’ soybean cultivar with high yield and abiotic stress resistance traits

    Technology Transfer Automated Retrieval System (TEKTRAN)

    ‘N8002’ is a conventional maturity-group (MG) VIII soybean [Glycine max (L.) Merr.] cultivar released by the USDA-ARS in September 2015. N8002 is the first North American MG VIII soybean cultivar exhibiting drought resistance traits and high yield potential. It is the first U.S. release derived from...

  15. ‘Caro-Tex 312’, a high yielding, orange-fruited, Habanero-type, F1 hybrid pepper

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Agricultural Research Service of the U. S. Department of Agriculture and the College of Agriculture and Life Sciences of Texas A&M University have released the high yielding, orange-fruited, Habanero-type, F1 hybrid pepper cultivar CaroTex-312. CaroTex-312 is the result of an F1 cross made at C...

  16. Heat Shock Transcriptional Responses in an MC-Producing Cyanobacterium (Planktothrix agardhii) and Its MC-Deficient Mutant under High Light Conditions

    PubMed Central

    Tran, Thi Du Chi; Bernard, Cecile; Ammar, Myriam; Chaouch, Soraya; Comte, Katia

    2013-01-01

    Microcystins (MCs) are the most commonly-reported hepatotoxins produced by various cyanobacterial taxa in fresh waters to constitute a potential threat to human and animal health. The biological role of MCs in the producer organisms is not known, and it would be very useful to understand the driving force behind the toxin production. Recent studies have suggested that MCs may have a protective function in cells facing environmental stress. Following this starting premise, we speculate that under adverse conditions the expression of stress-related genes coding for Heat Shock Proteins (Hsp) might be different in an MC-producing strain and its MC-deficient mutant. We therefore used RT-qPCR to compare the expression of 13 hsp genes of an MC-producing strain of Planktothrix agardhii (CYA126/8) and its MC-deficient ΔmcyD mutant over different periods of exposure to high light stress (HL). Three reference genes (RGs) were selected from six candidates to normalize the RT-qPCR data. Of these three RGs (rsh, rpoD, and gltA), gltA is used here for the first time as an RG in prokaryotes. Under HL stress, five genes were found to be strongly up-regulated in both strains (htpG, dnaK, hspA, groES, and groEL). Unexpectedly, we found that the MC-producing wild type strain accumulated higher levels of htpG and dnaK transcripts in response to HL stress than the MC-deficient mutant. In addition, a significant increase in the mcyE transcript was detected in the mutant, suggesting that MCs are required under HL conditions. We discuss several possible roles of MCs in the response to HL stress through their possible involvement in the protective mechanisms of the cells. PMID:24023831

  17. The relation of the yield stress of high-pressure anvils to the pressure attained at yielding and the ultimate attainable pressure

    NASA Technical Reports Server (NTRS)

    Panda, P. C.; Ruoff, A. L.

    1979-01-01

    A sensitive microprofilometer was used to determine the onset of yielding in the anvils of a supported opposed anvil device for the case of 3% cobalt-cemented tungsten carbide as the anvil material. In addition, it is shown how the commencement of yielding in boron carbide pistons, the yield strength being known, can be used to obtain the transition pressure to a conducting phase in gallium phosphide. The transition pressures of bismuth and gallium phosphide are obtained and it is found that these transitions are extremely close to the maximum attainable pressure in, respectively, a maraging steel and a 3% cobalt-cemented tungsten carbide.

  18. Anisotropic yielding of rocks at high temperatures and pressures; Annual Progress Report, 1988-1989

    SciTech Connect

    Kronenberg, A.K.; Russell, J.E.; Carter, N.L.

    1989-11-30

    The experimental results we have obtained on Four-Mile gneiss have demonstrated that the yield behavior of quartzo-feldspathic rocks containing only a small percentage (10%) of mica can be markedly anisotropic, provided the mica minerals exhibit a strong crystallographic preferred orientation. Samples of gneiss oriented such that resolved shear stresses on the foliation plane are large are considerably weaker than granites of similar grain size and composition, and this weakness is attributed to enhanced nucleation of microcracks in quartz and feldspar adjacent to mica grains that are suitably oriented for slip. We expect the yield behavior of rocks containing a higher proportion of phyllosilicates to be influenced by the strongly anisotropic nature of these minerals as well, although the strengths, temperature and pressure dependencies, and flow-controlling mechanisms in such rocks may be significantly different.

  19. A high-yielding, strictly regioselective prebiotic purine nucleoside formation pathway.

    PubMed

    Becker, Sidney; Thoma, Ines; Deutsch, Amrei; Gehrke, Tim; Mayer, Peter; Zipse, Hendrik; Carell, Thomas

    2016-05-13

    The origin of life is believed to have started with prebiotic molecules reacting along unidentified pathways to produce key molecules such as nucleosides. To date, a single prebiotic pathway to purine nucleosides had been proposed. It is considered to be inefficient due to missing regioselectivity and low yields. We report that the condensation of formamidopyrimidines (FaPys) with sugars provides the natural N-9 nucleosides with extreme regioselectivity and in good yields (60%). The FaPys are available from formic acid and aminopyrimidines, which are in turn available from prebiotic molecules that were also detected during the Rosetta comet mission. This nucleoside formation pathway can be fused to sugar-forming reactions to produce pentosides, providing a plausible scenario of how purine nucleosides may have formed under prebiotic conditions. PMID:27174989

  20. Gallium loading of gold seed for high yield of patterned GaAs nanowires

    SciTech Connect

    Boulanger, J. P.; Chia, A. C. E.; LaPierre, R. R.

    2014-08-25

    A method is presented for maximizing the yield and crystal phase purity of vertically aligned Au-assisted GaAs nanowires grown with an SiO{sub x} selective area epitaxy mask on GaAs (111)B substrates. The nanowires were grown by the vapor-liquid-solid (VLS) method in a gas source molecular beam epitaxy system. During annealing, Au VLS seeds will alloy with the underlying GaAs substrate and collect beneath the SiO{sub x} mask layer. This behavior is detrimental to obtaining vertically aligned, epitaxial nanowire growth. To circumvent this issue, Au droplets were pre-filled with Ga assuring vertical yields in excess of 99%.

  1. Advanced scheme for high-yield laser driven proton-boron fusion reaction

    NASA Astrophysics Data System (ADS)

    Margarone, D.; Picciotto, A.; Velyhan, A.; Krasa, J.; Kucharik, M.; Morrissey, M.; Mangione, A.; Szydlowsky, A.; Malinowska, A.; Bertuccio, G.; Shi, Y.; Crivellari, M.; Ullschmied, J.; Bellutti, P.; Korn, G.

    2015-02-01

    A low contrast nanosecond laser pulse with relatively low intensity (3 × 1016 W cm-2) was used to enhance the yield of induced nuclear reactions in advanced solid targets. In particular the "ultraclean" proton-boron fusion reaction, producing energetic alpha-particles without neutron generation, was chosen. A spatially well-defined layer of boron dopants in a hydrogen-enriched silicon substrate was used as target. The combination of the specific target geometry and the laser pulse temporal shape allowed enhancing the yield of alpha-particles up to 109 per steradian, i.e 100 times higher than previous experimental achievements. Moreover the alpha particle stream presented a clearly peaked angular and energy distribution, which make this secondary source attractive for potential applications. This result can be ascribed to the interaction of the long laser pre-pulse with the target and to the optimal target geometry and composition.

  2. Fast and high light yield scintillation in the Ga2O3 semiconductor material

    NASA Astrophysics Data System (ADS)

    Yanagida, Takayuki; Okada, Go; Kato, Takumi; Nakauchi, Daisuke; Yanagida, Satoko

    2016-04-01

    We report the distinct scintillation properties of the well-known Ga2O3 semiconductor material. Under UV excitation, the photoluminescence (PL) emission peak appeared near a wavelength of 380 nm with a quantum yield of 6%, and fast decays of 8 and 793 ns were observed. In contrast, the X-ray-induced scintillation spectrum showed an intense emission band near a wavelength of 380 nm, whose decay curve was reproduced using two exponential decay components with time constants of 8 and 977 ns. The pulse height spectrum of 137Cs γ-rays measured using Ga2O3 showed a clear photoabsorption peak with a light yield of 15000 ± 1500 photons/MeV.

  3. High quantum yield ZnO quantum dots synthesizing via an ultrasonication microreactor method.

    PubMed

    Yang, Weimin; Yang, Huafang; Ding, Wenhao; Zhang, Bing; Zhang, Le; Wang, Lixi; Yu, Mingxun; Zhang, Qitu

    2016-11-01

    Green emission ZnO quantum dots were synthesized by an ultrasonic microreactor. Ultrasonic radiation brought bubbles through ultrasonic cavitation. These bubbles built microreactor inside the microreactor. The photoluminescence properties of ZnO quantum dots synthesized with different flow rate, ultrasonic power and temperature were discussed. Flow rate, ultrasonic power and temperature would influence the type and quantity of defects in ZnO quantum dots. The sizes of ZnO quantum dots would be controlled by those conditions as well. Flow rate affected the reaction time. With the increasing of flow rate, the sizes of ZnO quantum dots decreased and the quantum yields first increased then decreased. Ultrasonic power changed the ultrasonic cavitation intensity, which affected the reaction energy and the separation of the solution. With the increasing of ultrasonic power, sizes of ZnO quantum dots first decreased then increased, while the quantum yields kept increasing. The effect of ultrasonic temperature on the photoluminescence properties of ZnO quantum dots was influenced by the flow rate. Different flow rate related to opposite changing trend. Moreover, the quantum yields of ZnO QDs synthesized by ultrasonic microreactor could reach 64.7%, which is higher than those synthesized only under ultrasonic radiation or only by microreactor. PMID:27245962

  4. Spontaneous high-yield hydrogen production from cellulosic materials and water catalyzed by enzyme cocktail

    SciTech Connect

    Ye, Xinhao; Wang, Yiran; Hopkins, Robert C.; Adams, Michael W. W.; Evans, Barbara R; Mielenz, Jonathan R; Zhang, Y.-H. Percival

    2009-01-01

    Carbon-neutral hydrogen gas is a compelling energy carrier, especially for the transportation section. Low-cost hydrogen can be produced from abundant renewable lignocellulosic biomass through a number of methods employing chemical catalysis, biocatalysis or a combination of both, but these technologies suffer from low hydrogen yields (well below the theoretical yield of 12 H2 per glucose), undesired side-products and/or required severe reaction conditions. Here we present a novel in vitro synthetic biology approach for producing near theoretical hydrogen yields from cellulosic materials (cellodextrins) and water at 32oC and 1 atm. These non-natural catabolic pathways containing up to 14 enzymes and one coenzyme degrade cellodextrins initially to glucose-1-phosphate and eventually to CO2, split water and finally release the chemical energy in the form of hydrogen gas. Up to 11.2 H2 per anhydroglucose was produced in a batch reaction. This spontaneous endothermic reaction is driven by entropy gain, suggesting that the thermal energy is adsorbed for generating more chemical energy (hydrogen gas) than that in cellodextrins, i.e., output/input of chemical energy > 1, with an input of ambient-temperature thermal energy.

  5. High quantum yield ZnO quantum dots synthesizing via an ultrasonication microreactor method.

    PubMed

    Yang, Weimin; Yang, Huafang; Ding, Wenhao; Zhang, Bing; Zhang, Le; Wang, Lixi; Yu, Mingxun; Zhang, Qitu

    2016-11-01

    Green emission ZnO quantum dots were synthesized by an ultrasonic microreactor. Ultrasonic radiation brought bubbles through ultrasonic cavitation. These bubbles built microreactor inside the microreactor. The photoluminescence properties of ZnO quantum dots synthesized with different flow rate, ultrasonic power and temperature were discussed. Flow rate, ultrasonic power and temperature would influence the type and quantity of defects in ZnO quantum dots. The sizes of ZnO quantum dots would be controlled by those conditions as well. Flow rate affected the reaction time. With the increasing of flow rate, the sizes of ZnO quantum dots decreased and the quantum yields first increased then decreased. Ultrasonic power changed the ultrasonic cavitation intensity, which affected the reaction energy and the separation of the solution. With the increasing of ultrasonic power, sizes of ZnO quantum dots first decreased then increased, while the quantum yields kept increasing. The effect of ultrasonic temperature on the photoluminescence properties of ZnO quantum dots was influenced by the flow rate. Different flow rate related to opposite changing trend. Moreover, the quantum yields of ZnO QDs synthesized by ultrasonic microreactor could reach 64.7%, which is higher than those synthesized only under ultrasonic radiation or only by microreactor.

  6. crw1--A novel maize mutant highly susceptible to foliar damage by the western corn rootworm beetle.

    PubMed

    Venkata, Bala Puchakayala; Lauter, Nick; Li, Xu; Chapple, Clint; Krupke, Christian; Johal, Gurmukh; Moose, Stephen

    2013-01-01

    Western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is the most destructive insect pest of corn (Zea mays L.) in the United States. The adult WCR beetles derive their nourishment from multiple sources including corn pollen and silks as well as the pollen of alternate hosts. Conversely, the corn foliage is largely neglected as a food source by WCR beetles, leading to a perception of a passive interaction between the two. We report here a novel recessive mutation of corn that was identified and named after its foliar susceptibility to corn rootworm beetles (crw1). The crw1 mutant under field conditions was exceptionally susceptible to foliar damage by WCR beetles in an age-specific manner. It exhibits pleiotropic defects on cell wall biochemistry, morphology of leaf epidermal cells and lower structural integrity via differential accumulation of cell wall bound phenolic acids. These findings indicate that crw1 is perturbed in a pathway that was not previously ascribed to WCR susceptibility, as well as implying the presence of an active mechanism(s) deterring WCR beetles from devouring corn foliage. The discovery and characterization of this mutant provides a unique opportunity for genetic analysis of interactions between maize and adult WCR beetles and identify new strategies to control the spread and invasion of this destructive pest.

  7. crw1 - A Novel Maize Mutant Highly Susceptible to Foliar Damage by the Western Corn Rootworm Beetle

    PubMed Central

    Venkata, Bala Puchakayala; Lauter, Nick; Li, Xu; Chapple, Clint; Krupke, Christian; Johal, Gurmukh; Moose, Stephen

    2013-01-01

    Western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is the most destructive insect pest of corn (Zea mays L.) in the United States. The adult WCR beetles derive their nourishment from multiple sources including corn pollen and silks as well as the pollen of alternate hosts. Conversely, the corn foliage is largely neglected as a food source by WCR beetles, leading to a perception of a passive interaction between the two. We report here a novel recessive mutation of corn that was identified and named after its foliar susceptibility to corn rootworm beetles (crw1). The crw1 mutant under field conditions was exceptionally susceptible to foliar damage by WCR beetles in an age-specific manner. It exhibits pleiotropic defects on cell wall biochemistry, morphology of leaf epidermal cells and lower structural integrity via differential accumulation of cell wall bound phenolic acids. These findings indicate that crw1 is perturbed in a pathway that was not previously ascribed to WCR susceptibility, as well as implying the presence of an active mechanism(s) deterring WCR beetles from devouring corn foliage. The discovery and characterization of this mutant provides a unique opportunity for genetic analysis of interactions between maize and adult WCR beetles and identify new strategies to control the spread and invasion of this destructive pest. PMID:23951124

  8. Characterization of a beta-glucosidase produced by a high-specific growth-rate mutant of Cellulomonas flavigena.

    PubMed

    Barrera-Islas, Gaspar A; Ramos-Valdivia, Ana C; Salgado, Luis M; Ponce-Noyola, Teresa

    2007-04-01

    The mutant strain PN-120 of Cellulomonas flavigena produces a ss-glucosidase that is 10-fold more active than the corresponding enzyme isolated from the parental strain. These enzymes were partially purified through Q Sepharose and Bio-Gel filtration. A single protein band was detected on polyacrylamide-gel electrophoresis/zymogram using 4-methylumbelliferyl-beta-D-glucoside. On sodium dodecyl sulfate-PAGE, the enzyme displayed three protein bands, suggesting that in C. flavigena the enzyme is oligomeric with a molecular mass of 210 kDa. On purification, the specific activity of ss-glucosidase isolated from PN-120 was increased 16-fold and showed three times more affinity for cellobiose than the enzyme of the parental strain; nevertheless, the optimum pH and temperature were similar for both enzymes. The kinetic parameters suggested that the increase in the activity of the enzyme, from the mutant strain, was caused by a mutation that affects the catalytic site of the enzyme. The partial amino-acid sequence of the isolated enzyme confirmed that it is a beta-glucosidase because of its homology with other beta-glucosidases produced by cellulolytic bacteria and fungi.

  9. Exogenously Applied Plant Growth Regulators Enhance the Morpho-Physiological Growth and Yield of Rice under High Temperature.

    PubMed

    Fahad, Shah; Hussain, Saddam; Saud, Shah; Hassan, Shah; Ihsan, Zahid; Shah, Adnan N; Wu, Chao; Yousaf, Muhammad; Nasim, Wajid; Alharby, Hesham; Alghabari, Fahad; Huang, Jianliang

    2016-01-01

    A 2-year experiment was conducted to ascertain the effects of exogenously applied plant growth regulators (PGR) on rice growth and yield attributes under high day (HDT) and high night temperature (HNT). Two rice cultivars (IR-64 and Huanghuazhan) were subjected to temperature treatments in controlled growth chambers and four different combinations of ascorbic acid (Vc), alpha-tocopherol (Ve), brassinosteroids (Br), methyl jasmonates (MeJA), and triazoles (Tr) were applied. High temperature severely affected rice morphology, and also reduced leaf area, above-, and below-ground biomass, photosynthesis, and water use efficiency, while increased the leaf water potential of both rice cultivars. Grain yield and its related attributes except number of panicles, were reduced under high temperature. The HDT posed more negative effects on rice physiological attributes, while HNT was more detrimental for grain formation and yield. The Huanghuazhan performed better than IR-64 under high temperature stress with better growth and higher grain yield. Exogenous application of PGRs was helpful in alleviating the adverse effects of high temperature. Among PGR combinations, the Vc+Ve+MejA+Br was the most effective treatment for both cultivars under high temperature stress. The highest grain production by Vc+Ve+MejA+Br treated plants was due to enhanced photosynthesis, spikelet fertility and grain filling, which compensated the adversities of high temperature stress. Taken together, these results will be of worth for further understanding the adaptation and survival mechanisms of rice to high temperature and will assist in developing heat-resistant rice germplasm in future.

  10. Exogenously Applied Plant Growth Regulators Enhance the Morpho-Physiological Growth and Yield of Rice under High Temperature.

    PubMed

    Fahad, Shah; Hussain, Saddam; Saud, Shah; Hassan, Shah; Ihsan, Zahid; Shah, Adnan N; Wu, Chao; Yousaf, Muhammad; Nasim, Wajid; Alharby, Hesham; Alghabari, Fahad; Huang, Jianliang

    2016-01-01

    A 2-year experiment was conducted to ascertain the effects of exogenously applied plant growth regulators (PGR) on rice growth and yield attributes under high day (HDT) and high night temperature (HNT). Two rice cultivars (IR-64 and Huanghuazhan) were subjected to temperature treatments in controlled growth chambers and four different combinations of ascorbic acid (Vc), alpha-tocopherol (Ve), brassinosteroids (Br), methyl jasmonates (MeJA), and triazoles (Tr) were applied. High temperature severely affected rice morphology, and also reduced leaf area, above-, and below-ground biomass, photosynthesis, and water use efficiency, while increased the leaf water potential of both rice cultivars. Grain yield and its related attributes except number of panicles, were reduced under high temperature. The HDT posed more negative effects on rice physiological attributes, while HNT was more detrimental for grain formation and yield. The Huanghuazhan performed better than IR-64 under high temperature stress with better growth and higher grain yield. Exogenous application of PGRs was helpful in alleviating the adverse effects of high temperature. Among PGR combinations, the Vc+Ve+MejA+Br was the most effective treatment for both cultivars under high temperature stress. The highest grain production by Vc+Ve+MejA+Br treated plants was due to enhanced photosynthesis, spikelet fertility and grain filling, which compensated the adversities of high temperature stress. Taken together, these results will be of worth for further understanding the adaptation and survival mechanisms of rice to high temperature and will assist in developing heat-resistant rice germplasm in future. PMID:27625658

  11. Exogenously Applied Plant Growth Regulators Enhance the Morpho-Physiological Growth and Yield of Rice under High Temperature

    PubMed Central

    Fahad, Shah; Hussain, Saddam; Saud, Shah; Hassan, Shah; Ihsan, Zahid; Shah, Adnan N.; Wu, Chao; Yousaf, Muhammad; Nasim, Wajid; Alharby, Hesham; Alghabari, Fahad; Huang, Jianliang

    2016-01-01

    A 2-year experiment was conducted to ascertain the effects of exogenously applied plant growth regulators (PGR) on rice growth and yield attributes under high day (HDT) and high night temperature (HNT). Two rice cultivars (IR-64 and Huanghuazhan) were subjected to temperature treatments in controlled growth chambers and four different combinations of ascorbic acid (Vc), alpha-tocopherol (Ve), brassinosteroids (Br), methyl jasmonates (MeJA), and triazoles (Tr) were applied. High temperature severely affected rice morphology, and also reduced leaf area, above-, and below-ground biomass, photosynthesis, and water use efficiency, while increased the leaf water potential of both rice cultivars. Grain yield and its related attributes except number of panicles, were reduced under high temperature. The HDT posed more negative effects on rice physiological attributes, while HNT was more detrimental for grain formation and yield. The Huanghuazhan performed better than IR-64 under high temperature stress with better growth and higher grain yield. Exogenous application of PGRs was helpful in alleviating the adverse effects of high temperature. Among PGR combinations, the Vc+Ve+MejA+Br was the most effective treatment for both cultivars under high temperature stress. The highest grain production by Vc+Ve+MejA+Br treated plants was due to enhanced photosynthesis, spikelet fertility and grain filling, which compensated the adversities of high temperature stress. Taken together, these results will be of worth for further understanding the adaptation and survival mechanisms of rice to high temperature and will assist in developing heat-resistant rice germplasm in future. PMID:27625658

  12. Exogenously Applied Plant Growth Regulators Enhance the Morpho-Physiological Growth and Yield of Rice under High Temperature

    PubMed Central

    Fahad, Shah; Hussain, Saddam; Saud, Shah; Hassan, Shah; Ihsan, Zahid; Shah, Adnan N.; Wu, Chao; Yousaf, Muhammad; Nasim, Wajid; Alharby, Hesham; Alghabari, Fahad; Huang, Jianliang

    2016-01-01

    A 2-year experiment was conducted to ascertain the effects of exogenously applied plant growth regulators (PGR) on rice growth and yield attributes under high day (HDT) and high night temperature (HNT). Two rice cultivars (IR-64 and Huanghuazhan) were subjected to temperature treatments in controlled growth chambers and four different combinations of ascorbic acid (Vc), alpha-tocopherol (Ve), brassinosteroids (Br), methyl jasmonates (MeJA), and triazoles (Tr) were applied. High temperature severely affected rice morphology, and also reduced leaf area, above-, and below-ground biomass, photosynthesis, and water use efficiency, while increased the leaf water potential of both rice cultivars. Grain yield and its related attributes except number of panicles, were reduced under high temperature. The HDT posed more negative effects on rice physiological attributes, while HNT was more detrimental for grain formation and yield. The Huanghuazhan performed better than IR-64 under high temperature stress with better growth and higher grain yield. Exogenous application of PGRs was helpful in alleviating the adverse effects of high temperature. Among PGR combinations, the Vc+Ve+MejA+Br was the most effective treatment for both cultivars under high temperature stress. The highest grain production by Vc+Ve+MejA+Br treated plants was due to enhanced photosynthesis, spikelet fertility and grain filling, which compensated the adversities of high temperature stress. Taken together, these results will be of worth for further understanding the adaptation and survival mechanisms of rice to high temperature and will assist in developing heat-resistant rice germplasm in future.

  13. Tuning of ZIF-Derived Carbon with High Activity, Nitrogen Functionality, and Yield - A Case for Superior CO2 Capture.

    PubMed

    Gadipelli, Srinivas; Guo, Zheng Xiao

    2015-06-22

    A highly effective and facile synthesis route is developed to create and tailor metal-decorated and nitrogen-functionalized active microporous carbon materials from ZIF-8. Clear metal- and pyrrolic-N-induced enhancements of the cyclic CO2 uptake capacities and binding energies are achieved, particularly at a much lower carbonization temperature of 700 °C than those often reported (1000 °C). The high-temperature carbonization can enhance the porosity but only at the expense of considerable losses of sample yield and metal and N functional sites. The findings are comparatively discussed with carbons derived from metal-organic frameworks (MOFs) reported previously. Furthermore, the porosity of the MOF-derived carbon is critically dependent on the structure of the precursor MOF and the crystal growth. The current strategy offers a new and effective route for the creation and tuning of highly active and functionalized carbon structures in high yields and with low energy consumption.

  14. Direct and efficient ethanol production from high-yielding rice using a Saccharomyces cerevisiae strain that express amylases.

    PubMed

    Yamada, Ryosuke; Yamakawa, Syun-Ichi; Tanaka, Tsutomu; Ogino, Chiaki; Fukuda, Hideki; Kondo, Akihiko

    2011-04-01

    Efficient ethanol producing yeast Saccharomyces cerevisiae cannot produce ethanol from raw starch directly. Thus the conventional ethanol production required expensive and complex process. In this study, we developed a direct and efficient ethanol production process from high-yielding rice harvested in Japan by using amylase expressing yeast without any pretreatment or addition of enzymes or nutrients. Ethanol productivity from high-yielding brown rice (1.1g/L/h) was about 5-fold higher than that obtained from purified raw corn starch (0.2g/L/h) when nutrients were added. Using an inoculum volume equivalent to 10% of the fermentation volume without any nutrient supplementation resulted in ethanol productivity and yield reaching 1.2g/L/h and 101%, respectively, in a 24-h period. High-yielding rice was demonstrated to be a suitable feedstock for bioethanol production. In addition, our polyploid amylase-expressing yeast was sufficiently robust to produce ethanol efficiently from real biomass. This is first report of direct ethanol production on real biomass using an amylase-expressing yeast strain without any pretreatment or commercial enzyme addition.

  15. Vineyard Yield Estimation Based on the Analysis of High Resolution Images Obtained with Artificial Illumination at Night

    PubMed Central

    Font, Davinia; Tresanchez, Marcel; Martínez, Dani; Moreno, Javier; Clotet, Eduard; Palacín, Jordi

    2015-01-01

    This paper presents a method for vineyard yield estimation based on the analysis of high-resolution images obtained with artificial illumination at night. First, this paper assesses different pixel-based segmentation methods in order to detect reddish grapes: threshold based, Mahalanobis distance, Bayesian classifier, linear color model segmentation and histogram segmentation, in order to obtain the best estimation of the area of the clusters of grapes in this illumination conditions. The color spaces tested were the original RGB and the Hue-Saturation-Value (HSV). The best segmentation method in the case of a non-occluded reddish table-grape variety was the threshold segmentation applied to the H layer, with an estimation error in the area of 13.55%, improved up to 10.01% by morphological filtering. Secondly, after segmentation, two procedures for yield estimation based on a previous calibration procedure have been proposed: (1) the number of pixels corresponding to a cluster of grapes is computed and converted directly into a yield estimate; and (2) the area of a cluster of grapes is converted into a volume by means of a solid of revolution, and this volume is converted into a yield estimate; the yield errors obtained were 16% and −17%, respectively. PMID:25860071

  16. Off-shell effects for the reaction pp{yields}{pi}d at high energies

    SciTech Connect

    Lee, T.S.H.; Locher, M.P.; Lu, Y.

    1995-08-01

    The reaction pp {yields} {pi}d is studied in a relativistic meson rescattering model. For 1.3 < T{sub p} < 2.4 GeV, the differential cross section and the asymmetry are calculated and compared to experiment. The model introduces simple form factors for the leading {pi}N partial waves, which depend on the virtuality of the exchanged {pi} and {rho} mesons. All remaining input is derived from experimental constraints. The data can be described by energy-independent form factors. The asymmetries are sensitive to pp distortion factors and further details of the model. A paper describing our results was published.

  17. High-yield resin fractionation using a liquid/liquid centrifuge

    NASA Astrophysics Data System (ADS)

    Wanat, Stan F.; Rahman, M. D.; Narasimhan, Balaji; McKenzie, Douglas S.; Cook, Michelle M.

    2000-06-01

    Resins used in photoresist manufacturing are often relatively expensive once processing steps (fractionation e.g.) and yield losses are factored into the net cost. We have previously reported on the merits of using an economically more attractive fractionation process using a liquid/liquid centrifuge. Further refinements of this method indicate that waste streams could be reduced by recycling the extractant phase and that lower molecular weight fractions removed from the starting resin might be used in making other resist ingredients [speed enhancers, photoactive compound (PAC) backbones e.g.]. Both of these improvements would reduce the overall manufacturing costs of making resist raw materials and the final products made with them.

  18. High Resolution Structure of Deinococcus Bacteriophytochrome Yields New Insights into Phytochrome Architecture and Evolution

    SciTech Connect

    Wagner, Jeremiah R.; Zhang, Junrui; Brunzelle, Joseph S.; Vierstra, Richard D.; Forest, Katrina T.

    2010-03-08

    Phytochromes are red/far red light photochromic photoreceptors that direct many photosensory behaviors in the bacterial, fungal, and plant kingdoms. They consist of an N-terminal domain that covalently binds a bilin chromophore and a C-terminal region that transmits the light signal, often through a histidine kinase relay. Using x-ray crystallography, we recently solved the first three-dimensional structure of a phytochrome, using the chromophore-binding domain of Deinococcus radiodurans bacterial phytochrome assembled with its chromophore, biliverdin IX{alpha}. Now, by engineering the crystallization interface, we have achieved a significantly higher resolution model. This 1.45 {angstrom} resolution structure helps identify an extensive buried surface between crystal symmetry mates that may promote dimerization in vivo. It also reveals that upon ligation of the C3{sup 2} carbon of biliverdin to Cys{sup 24}, the chromophore A-ring assumes a chiral center at C2, thus becoming 2(R),3(E)-phytochromobilin, a chemistry more similar to that proposed for the attached chromophores of cyanobacterial and plant phytochromes than previously appreciated. The evolution of bacterial phytochromes to those found in cyanobacteria and higher plants must have involved greater fitness using more reduced bilins, such as phycocyanobilin, combined with a switch of the attachment site from a cysteine near the N terminus to one conserved within the cGMP phosphodiesterase/adenyl cyclase/FhlA domain. From analysis of site-directed mutants in the D. radiodurans phytochrome, we show that this bilin preference was partially driven by the change in binding site, which ultimately may have helped photosynthetic organisms optimize shade detection. Collectively, these three-dimensional structural results better clarify bilin/protein interactions and help explain how higher plant phytochromes evolved from prokaryotic progenitors.

  19. Electrochemical selection and characterization of a high current-generating Shewanella oneidensis mutant with altered cell-surface morphology and biofilm-related gene expression

    PubMed Central

    2014-01-01

    Background Shewanella oneidensis MR-1 exhibits extracellular electron transfer (EET) activity that is influenced by various cellular components, including outer-membrane cytochromes, cell-surface polysaccharides (CPS), and regulatory proteins. Here, a random transposon-insertion mutant library of S. oneidensis MR-1 was screened after extended cultivation in electrochemical cells (ECs) with a working electrode poised at +0.2 V (vs. Ag/AgCl) to isolate mutants that adapted to electrode-respiring conditions and identify as-yet-unknown EET-related factors. Results Several mutants isolated from the enrichment culture exhibited rough morphology and extraordinarily large colonies on agar plates compared to wild-type MR-1. One of the isolated mutants, designated strain EC-2, produced 90% higher electric current than wild-type MR-1 in ECs and was found to have a transposon inserted in the SO_1860 (uvrY) gene, which encodes a DNA-binding response regulator of the BarA/UvrY two-component regulatory system. However, an in-frame deletion mutant of SO_1860 (∆SO_1860) did not exhibit a similar level of current generation as that of EC-2, suggesting that the enhanced current-generating capability of EC-2 was not simply due to the disruption of SO_1860. In both EC-2 and ∆SO_1860, the transcription of genes related to CPS synthesis was decreased compared to wild-type MR-1, suggesting that CPS negatively affects current generation. In addition, transcriptome analyses revealed that a number of genes, including those involved in biofilm formation, were differentially expressed in EC-2 compared to those in ∆SO_1860. Conclusions The present results indicate that the altered expression of the genes related to CPS biosynthesis and biofilm formation is associated with the distinct morphotype and high current-generating capability of strain EC-2, suggesting an important role of these genes in determining the EET activity of S. oneidensis. PMID:25028134

  20. High temperature pre-digestion of corn stover biomass for improved product yields

    DOE PAGESBeta

    Brunecky, Roman; Hobdey, Sarah E.; Taylor, Larry E.; Tao, Ling; Tucker, Melvin P.; Himmel, Michael E.; Decker, Stephen R.

    2014-12-03

    Introduction: The efficient conversion of lignocellulosic feedstocks remains a key step in the commercialization of biofuels. One of the barriers to cost-effective conversion of lignocellulosic biomass to sugars remains the enzymatic saccharification process step. Here, we describe a novel hybrid processing approach comprising enzymatic pre-digestion with newly characterized hyperthermophilic enzyme cocktails followed by conventional saccharification with commercial enzyme preparations. Dilute acid pretreated corn stover was subjected to this new procedure to test its efficacy. Thermal tolerant enzymes from Acidothermus cellulolyticus and Caldicellulosiruptor bescii were used to pre-digest pretreated biomass at elevated temperatures prior to saccharification by the commercial cellulase formulation.more » Results: We report that pre-digestion of biomass with these enzymes at elevated temperatures prior to addition of the commercial cellulase formulation increased conversion rates and yields when compared to commercial cellulase formulation alone under low solids conditions. In conclusion, Our results demonstrating improvements in rates and yields of conversion point the way forward for hybrid biomass conversion schemes utilizing catalytic amounts of hyperthermophilic enzymes.« less

  1. High temperature pre-digestion of corn stover biomass for improved product yields

    SciTech Connect

    Brunecky, Roman; Hobdey, Sarah E.; Taylor, Larry E.; Tao, Ling; Tucker, Melvin P.; Himmel, Michael E.; Decker, Stephen R.

    2014-12-03

    Introduction: The efficient conversion of lignocellulosic feedstocks remains a key step in the commercialization of biofuels. One of the barriers to cost-effective conversion of lignocellulosic biomass to sugars remains the enzymatic saccharification process step. Here, we describe a novel hybrid processing approach comprising enzymatic pre-digestion with newly characterized hyperthermophilic enzyme cocktails followed by conventional saccharification with commercial enzyme preparations. Dilute acid pretreated corn stover was subjected to this new procedure to test its efficacy. Thermal tolerant enzymes from Acidothermus cellulolyticus and Caldicellulosiruptor bescii were used to pre-digest pretreated biomass at elevated temperatures prior to saccharification by the commercial cellulase formulation. Results: We report that pre-digestion of biomass with these enzymes at elevated temperatures prior to addition of the commercial cellulase formulation increased conversion rates and yields when compared to commercial cellulase formulation alone under low solids conditions. In conclusion, Our results demonstrating improvements in rates and yields of conversion point the way forward for hybrid biomass conversion schemes utilizing catalytic amounts of hyperthermophilic enzymes.

  2. New Procedures for Purification of l-Asparaginase with High Yield from Escherichia coli

    PubMed Central

    Roberts, Joseph; Burson, Gene; Hill, Joseph M.

    1968-01-01

    l-Asparaginase is now known to be a potent antineoplastic agent in animals and has given complete remission in some human leukemias. Extensive clinical trials of this enzyme, however, were not possible in the past because of inadequate production of this substance. We have developed practical procedures for producing l-asparaginase in yields of sufficient quantity and purity for more extensive clinical evaluation. The nutritional requirements for optimal production of biologically active l-asparaginase by a strain of Escherichia coli have been ascertained. The highest yields of enzyme were obtained when cells were grown aerobically in a corn steep medium. Good enzyme production was associated with media containing l-glutamic acid, l-methionine, and lactic acid. The addition of glucose to the medium, however, resulted in depressed production of l-asparaginase. Sodium ion appeared to suppress l-asparaginase production. With the procedure described for isolation of biologically active l-asparaginase from E. coli, stable l-asparaginase preparations with a specific activity of 620 IU per mg of protein (1,240-fold purification with 40% total recovery) were obtained. PMID:4970225

  3. High yields of fatty acid and neutral lipid production from cassava bagasse hydrolysate (CBH) by heterotrophic Chlorella protothecoides.

    PubMed

    Chen, Junhui; Liu, Xiaoguang; Wei, Dong; Chen, Gu

    2015-09-01

    The fermentation process for high yields of fatty acid and neutral lipid production from cassava bagasse hydrolysate (CBH) was developed by heterotrophic Chlorella protothecoides. An efficient single-step enzymatic hydrolysis of cassava bagasse (CB) by cellulase was firstly developed to produce >30 g/L of reducing sugars. The concentrated CBH was subsequently applied in a batch culture, producing 7.9 g/L of dry biomass with yield of 0.44 g/g reducing sugar and 34.3 wt% of fatty acids and 48.6 wt% of neutral lipids. Furthermore, fed-batch fermentation using CBH achieved higher yields of fatty acids (41.0 wt% and a titer of 5.83 g/L) and neutral lipids (58.4 wt% and yield of 0.22 g/g reducing sugar). Additionally, the fatty acid profile analysis showed that the intercellular lipid was suitable to prepare high-quality biodiesel. This study demonstrated the feasibility of using CBH as low-cost feedstock to produce crude algal oil for sustainable biodiesel production.

  4. Toward Self-Assembled Plasmonic Devices: High-Yield Arrangement of Gold Nanoparticles on DNA Origami Templates.

    PubMed

    Gür, Fatih N; Schwarz, Friedrich W; Ye, Jingjing; Diez, Stefan; Schmidt, Thorsten L

    2016-05-24

    Plasmonic structures allow the manipulation of light with materials that are smaller than the optical wavelength. Such structures can consist of plasmonically active metal nanoparticles and can be fabricated through scalable bottom-up self-assembly on DNA origami templates. To produce functional devices, the precise and high-yield arrangement of each of the nanoparticles on a structure is of vital importance as the absence of a single particle can destroy the functionality of the entire device. Nevertheless, the parameters influencing the yield of the multistep assembly process are still poorly understood. To overcome this deficiency, we employed a test system consisting of a tubular six-helix bundle DNA origami with binding sites for eight oligonucleotide-functionalized gold nanoparticles. We systematically studied the assembly yield as a function of a wide range of parameters such as ionic strength, stoichiometric ratio, oligonucleotide linker chemistry, and assembly kinetics by an automated high-throughput analysis of electron micrographs of the formed heterocomplexes. Our optimized protocols enable particle placement yields up to 98.7% and promise the reliable production of sophisticated DNA-based multiparticle plasmonic devices for applications in photonics, optoelectronics, and nanomedicine. PMID:27159647

  5. Optimizing Hill Seeding Density for High-Yielding Hybrid Rice in a Single Rice Cropping System in South China

    PubMed Central

    Wang, Danying; Chen, Song; Wang, Zaiman; Ji, Chenglin; Xu, Chunmei; Zhang, Xiufu; Chauhan, Bhagirath Singh

    2014-01-01

    Mechanical hill direct seeding of hybrid rice could be the way to solve the problems of high seeding rates and uneven plant establishment now faced in direct seeded rice; however, it is not clear what the optimum hill seeding density should be for high-yielding hybrid rice in the single-season rice production system. Experiments were conducted in 2010 and 2011 to determine the effects of hill seeding density (25 cm×15 cm, 25 cm×17 cm, 25 cm×19 cm, 25 cm×21 cm, and 25 cm×23 cm; three to five seeds per hill) on plant growth and grain yield of a hybrid variety, Nei2you6, in two fields with different fertility (soil fertility 1 and 2). In addition, in 2012 and 2013, comparisons among mechanical hill seeding, broadcasting, and transplanting were conducted with three hybrid varieties to evaluate the optimum seeding density. With increases in seeding spacing from 25 cm×15 cm to 25 cm×23 cm, productive tillers per hill increased by 34.2% and 50.0% in soil fertility 1 and 2. Panicles per m2 declined with increases in seeding spacing in soil fertility 1. In soil fertility 2, no difference in panicles per m2 was found at spacing ranging from 25 cm×17 cm to 25 cm×23 cm, while decreases in the area of the top three leaves and aboveground dry weight per shoot at flowering were observed. Grain yield was the maximum at 25 cm×17 cm spacing in both soil fertility fields. Our results suggest that a seeding density of 25 cm×17 cm was suitable for high-yielding hybrid rice. These results were verified through on-farm demonstration experiments, in which mechanical hill-seeded rice at this density had equal or higher grain yield than transplanted rice. PMID:25290342

  6. Registration of ‘highpro1’ soybean with high protein and high yield developed from a North x South cross

    Technology Transfer Automated Retrieval System (TEKTRAN)

    ‘Highpro1’ soybean [Glycine max (L.) Merr.]) (Reg. no. , PI ) was developed and jointly released by the USDA-Agricultural Research Service and The Ohio Agricultural Research and Development Center (OARDC) in Wooster, OH as a high-protein high-yielding maturity group (MG) III (rel...

  7. Production of aggregation prone human interferon gamma and its mutant in highly soluble and biologically active form by SUMO fusion technology.

    PubMed

    Tileva, M; Krachmarova, E; Ivanov, I; Maskos, K; Nacheva, G

    2016-01-01

    The Escherichia coli expression system is a preferable choice for production of recombinant proteins. A disadvantage of this system is the target protein aggregation in "inclusion bodies" (IBs) that further requires solubilisation and refolding, which is crucial for the properties and the yield of the final product. In order to prevent aggregation, SUMO fusion tag technology has been successfully applied for expression of eukaryotic proteins, including human interferon gamma (hIFNγ) that was reported, however, with no satisfactory biological activity. We modified this methodology for expression and purification of both the wild type hIFNγ and an extremely prone to aggregation mutant hIFNγ-K88Q, whose recovery from IBs showed to be ineffective upon numerous conditions. By expression of the N-terminal His-SUMO fusion proteins in the E. coli strain BL21(DE3)pG-KJE8, co-expressing two chaperone systems, at 24 °C a significant increase in solubility of both target proteins (1.5-fold for hIFNγ and 8-fold for K88Q) was achieved. Two-step chromatography (affinity and ion-exchange) with on-dialysis His-SUMO-tag cleavage was applied for protein purification that yielded 6.0-7.0mg/g wet biomass for both proteins with >95% purity and native N-termini. The optimised protocol led to increased yields from 5.5 times for hIFNγ up to 100 times for K88Q in comparison to their isolation from IBs. Purified hIFNγ showed preserved thermal stability and antiproliferative activity corresponding to that of the native reference sample (3 × 10(7)IU/mg). The developed methodology represents an optimised procedure that can be successfully applied for large scale expression and purification of aggregation-prone proteins in soluble native form.

  8. Resource-Efficient High-Yield Ionothermal Synthesis of Microcrystalline Cu3-xP.

    PubMed

    Wolff, Alexander; Pallmann, Julia; Boucher, Richard; Weiz, Alexander; Brunner, Eike; Doert, Thomas; Ruck, Michael

    2016-09-01

    Polycrystalline Cu3-xP was successfully synthesized in different ionic liquids comprising imidazolium and phosphonium cations. The reaction of elemental copper and red phosphorus in trihexyltetradecylphosphonium chloride at 200 °C led to single-phase Cu3-xP (x = 0.05) within 24 h with a quantitative yield (99%). Liquid-state nuclear magnetic resonance spectroscopy of the ionic liquids revealed degeneration of the imidazolium cations under the synthesis conditions, while phosphonium cations remain stable. The solid products were characterized with X-ray powder diffraction, scanning electron microscopy, energy-dispersive X-ray spectroscopy, solid-state nuclear magnetic resonance spectroscopy, and elemental analysis. A reinvestigation of the electronic transport properties of Cu2.95(4)P showed metallic behavior for the bulk material. The formation of CuP2 during the synthesis of phosphorus-rich Cu3-xP (x ≥ 0.1) was observed. PMID:27559933

  9. Synthesis of a high-yield activated carbon by air gasification of macadamia nut shell charcoal

    SciTech Connect

    Dai, X.; Antal, M.J. Jr.

    1999-09-01

    Macadamia nut shell charcoal was heated in an inert environment to temperatures above 1000 K (carbonized), reacted with oxygen (Po{sub 2} = 2.68--11.3 kPa) at temperatures between 525 and 586 K (oxygenated), and heated again in an inert environment to temperatures above 1000 K (activated) to produce an activated carbon. Carbons produced by this process possess surface areas and iodine numbers in the range of 400--550. Overall yields of these carbons (based on the dry, raw macadamia nut shell feed) ranged from 24 to 30 wt %. Under the conditions employed in this work, the rates of chemisorption and gasification were not mass transfer limited. Initially, the gasification reaction was first-order with respect to oxygen concentration but became independent of oxygen concentration as the surface sites of the carbon became saturated with oxygen.

  10. High-yield total synthesis of (-)-strictinin through intramolecular coupling of gallates.

    PubMed

    Michihata, Naoki; Kaneko, Yuki; Kasai, Yusuke; Tanigawa, Kotaro; Hirokane, Tsukasa; Higasa, Sho; Yamada, Hidetoshi

    2013-05-01

    This paper describes a total synthesis of (-)-strictinin, an ellagitannin that is 1-O-galloyl-4,6-O-(S)-hexahydroxydiphenoyl (HHDP)-β-D-glucose. In the study, total efficiency of the synthesis was improved to produce a 78% overall yield in 13 steps from D-glucose. In the synthesis, formation of the 4,6-(S)-HHDP bridge including the 11-membered bislactone ring was a key step, in which intramolecular aryl-aryl coupling was adopted. The coupling was oxidatively induced by CuCl2-n-BuNH2 with perfect control of the axial chirality, and the reaction conditions of this coupling were optimized thoroughly to achieve the quantitative formation of the bridge.

  11. High liquid fuel yielding biofuel processes and a roadmap for the future transportation

    NASA Astrophysics Data System (ADS)

    Singh, Navneet R.

    In a fossil-fuel deprived world when crude oil will be scarce and transportation need cannot be met with electricity and transportation liquid fuel must be produced, biomass derived liquid fuels can be a natural replacement. However, the carbon efficiency of the currently known biomass to liquid fuel conversion processes ranges from 35-40%, yielding 90 ethanol gallon equivalents (ege) per ton of biomass. This coupled with the fact that the efficiency at which solar energy is captured by biomass (<1%) is significantly lower than H 2 (10-27%) and electricity (20-42%), implies that sufficient land area is not available to meet the need for the entire transportation sector. To counter this dilemma, a number of processes have been proposed in this work: a hybrid hydrogen-carbon (H2CAR) process based on biomass gasification followed by the Fischer-Tropsch process such that 100% carbon efficiency is achieved yielding 330 ege/ton biomass using hydrogen derived from a carbon-free energy. The hydrogen requirement for the H2CAR process is 0.33 kg/liter of diesel. To decrease the hydrogen requirement associated with the H2CAR process, a hydrogen bio-oil (H2Bioil) process based on biomass fast-hydropyrolysis/hydrodeoxygenation is proposed which can achieve liquid fuel yield of 215 ege/ton consuming 0.11 kg hydrogen per liter of oil. Due to the lower hydrogen consumption of the H2Bioil process, synergistically integrated transition pathways are feasible where hot syngas derived from coal gasification (H2Bioil-C) or a natural gas reformer (H 2Bioil-NG) is used to supply the hydrogen and process heat for the biomass fast-hydropyrolysis/hydrodeoxygenation. Another off-shoot of the H2Bioil process is the H2Bioil-B process, where hydrogen required for the hydropyrolysis is obtained from gasification of a fraction of the biomass. H2Bioil-B achieves the highest liquid fuel yield (126-146 ege/ton of biomass) reported in the literature for any self-contained conversion of biomass to

  12. High-yield production of biosugars from Gracilaria verrucosa by acid and enzymatic hydrolysis processes.

    PubMed

    Kim, Se Won; Hong, Chae-Hwan; Jeon, Sung-Wan; Shin, Hyun-Jae

    2015-11-01

    Gracilaria verrucosa, the red alga, is a suitable feedstock for biosugar production. This study analyzes biosugar production by the hydrolysis of G. verrucosa conducted under various conditions (i.e., various acid concentrations, substrate concentrations, reaction times, and enzyme dosages). The acid hydrolysates of G. verrucosa yielded a total of 7.47g/L (37.4%) and 10.63g/L (21.26%) of reducing sugars under optimal small (30mL) and large laboratory-scale (1L) hydrolysis processes, respectively. Reducing sugar obtained from acid and enzymatic hydrolysates were 10% higher, with minimum by-products, than those reported in other studies. The mass balance for the small laboratory-scale process showed that the acid and enzymatic hydrolysates had a carbohydrate conversion of 57.2%. The mass balance approach to the entire hydrolysis process of red seaweed for biosugar production can be applied to other saccharification processes.

  13. High-Yielding Green Hydrothermal Synthesis of Ruthenium Nanoparticles and Their Characterization.

    PubMed

    Dikhtiarenko, Alla; Khainakov, Sergei A; Khaynakova, Olena; García, José R; Gimeno, José

    2016-06-01

    Using hydrothermal techniques, a novel synthetic approach to prepare ruthenium nanoparticles has been developed. At 180 degrees C and under autogenous pressure, starting from an aqueous solution of ruthenium trichloride, the method yielded nanoparticles whose form and size both depended on the reducing agent: sodium citrate (hexagonal shaped nanocrystals, 1-20 nm), ascorbic acid (spherical nanoparticles, 3-5 nm) and succinic acid (spherical nanoparticles, 1-120 nm). Depending on the reaction variables, the nature and concentration of partially reduced species determines the characteristics of the final products. HRTEM image analysis along with the simulation techniques were stabilized preferential growth of nanoparticles on specific directions. Ruthenium samples have been investigated by Temperature-Programmed Reduction (TPR) showing that the reduction temperature of nanoparticles is correlated to their nanocrystalline size. PMID:27427683

  14. High-yield production of a stable Vero cell-based vaccine candidate against the highly pathogenic avian influenza virus H5N1

    SciTech Connect

    Zhou, Fangye; Zhou, Jian; Ma, Lei; Song, Shaohui; Zhang, Xinwen; Li, Weidong; Jiang, Shude; Wang, Yue; Liao, Guoyang

    2012-05-18

    Highlights: Black-Right-Pointing-Pointer Vero cell-based HPAI H5N1 vaccine with stable high yield. Black-Right-Pointing-Pointer Stable high yield derived from the YNVa H3N2 backbone. Black-Right-Pointing-Pointer H5N1/YNVa has a similar safety and immunogenicity to H5N1delta. -- Abstract: Highly pathogenic avian influenza (HPAI) viruses pose a global pandemic threat, for which rapid large-scale vaccine production technology is critical for prevention and control. Because chickens are highly susceptible to HPAI viruses, the supply of chicken embryos for vaccine production might be depleted during a virus outbreak. Therefore, developing HPAI virus vaccines using other technologies is critical. Meeting vaccine demand using the Vero cell-based fermentation process has been hindered by low stability and yield. In this study, a Vero cell-based HPAI H5N1 vaccine candidate (H5N1/YNVa) with stable high yield was achieved by reassortment of the Vero-adapted (Va) high growth A/Yunnan/1/2005(H3N2) (YNVa) virus with the A/Anhui/1/2005(H5N1) attenuated influenza vaccine strain (H5N1delta) using the 6/2 method. The reassorted H5N1/YNVa vaccine maintained a high hemagglutination (HA) titer of 1024. Furthermore, H5N1/YNVa displayed low pathogenicity and uniform immunogenicity compared to that of the parent virus.

  15. Development and characterization of a high yield transportable pulsed neutron source with efficient and compact pulsed power system

    NASA Astrophysics Data System (ADS)

    Verma, Rishi; Mishra, Ekansh; Dhang, Prosenjit; Sagar, Karuna; Meena, Manraj; Shyam, Anurag

    2016-09-01

    The results of characterization experiments carried out on a newly developed dense plasma focus device based intense pulsed neutron source with efficient and compact pulsed power system are reported. Its high current sealed pseudospark switch based low inductance capacitor bank with maximum stored energy of ˜10 kJ is segregated into four modules of ˜2.5 kJ each and it cumulatively delivers peak current in the range of 400 kA-600 kA (corresponding to charging voltage range of 14 kV-18 kV) in a quarter time period of ˜2 μs. The neutron yield performance of this device has been optimized by discretely varying deuterium filling gas pressure in the range of 6 mbar-11 mbar at ˜17 kV/550 kA discharge. At ˜7 kJ/8.5 mbar operation, the average neutron yield has been measured to be in the order of ˜4 × 109 neutrons/pulse which is the highest ever reported neutron yield from a plasma focus device with the same stored energy. The average forward to radial anisotropy in neutron yield is found to be ˜2. The entire system is contained on a moveable trolley having dimensions 1.5 m × 1 m × 0.7 m and its operation and control (up to the distance of 25 m) are facilitated through optically isolated handheld remote console. The overall compactness of this system provides minimum proximity to small as well as large samples for irradiation. The major intended application objective of this high neutron yield dense plasma focus device development is to explore the feasibility of active neutron interrogation experiments by utilization of intense pulsed neutron sources.

  16. CO₂ enrichment can produce high red leaf lettuce yield while increasing most flavonoid glycoside and some caffeic acid derivative concentrations.

    PubMed

    Becker, Christine; Kläring, Hans-Peter

    2016-05-15

    Carbon dioxide (CO2) enrichment is a common practice in greenhouses to increase crop yields up to 30%. Yet, reports on the effect on foliar phenolic compounds vary. We studied the effect on two red leaf lettuce cultivars, grown for 25 days in growth chambers at CO2 concentrations of 200 or 1,000 ppm, with some plants exchanged between treatments after 11 days. As expected, head mass increased with higher CO2 concentration. Regression analysis, corrected for head mass, showed increased concentrations of most flavonoid glycosides at high CO2 concentrations while only some caffeic acid derivatives were increased, and not uniformly in both cultivars. Sugar concentrations increased with CO2 concentration. Generally, conditions in the 10 days before harvest determined concentrations. We suspect that phenolic compounds were mainly accumulated because plenty of precursors were available. The results indicate that CO2 enrichment can result in high yields of red leaf lettuce rich in phenolic compounds.

  17. CO₂ enrichment can produce high red leaf lettuce yield while increasing most flavonoid glycoside and some caffeic acid derivative concentrations.

    PubMed

    Becker, Christine; Kläring, Hans-Peter

    2016-05-15

    Carbon dioxide (CO2) enrichment is a common practice in greenhouses to increase crop yields up to 30%. Yet, reports on the effect on foliar phenolic compounds vary. We studied the effect on two red leaf lettuce cultivars, grown for 25 days in growth chambers at CO2 concentrations of 200 or 1,000 ppm, with some plants exchanged between treatments after 11 days. As expected, head mass increased with higher CO2 concentration. Regression analysis, corrected for head mass, showed increased concentrations of most flavonoid glycosides at high CO2 concentrations while only some caffeic acid derivatives were increased, and not uniformly in both cultivars. Sugar concentrations increased with CO2 concentration. Generally, conditions in the 10 days before harvest determined concentrations. We suspect that phenolic compounds were mainly accumulated because plenty of precursors were available. The results indicate that CO2 enrichment can result in high yields of red leaf lettuce rich in phenolic compounds. PMID:26776031

  18. Brucella abortusΔcydCΔcydD and ΔcydCΔpurD double-mutants are highly attenuated and confer long-term protective immunity against virulent Brucella abortus.

    PubMed

    Truong, Quang Lam; Cho, Youngjae; Park, Soyeon; Kim, Kiju; Hahn, Tae-Wook

    2016-01-01

    We constructed double deletion (ΔcydCΔcydD and ΔcydCΔpurD) mutants from virulent Brucella abortus biovar 1 field isolate (BA15) by deleting the genes encoding an ATP-binding cassette-type transporter (cydC and cydD genes) and a phosphoribosylamine-glycine ligase (purD). Both BA15ΔcydCΔcydD and BA15ΔcydCΔpurD double-mutants exhibited significant attenuation of virulence when assayed in murine macrophages or in BALB/c mice. Both double-mutants were readily cleared from spleens by 4 weeks post-inoculation even when inoculated at the dose of 10(8) CFU per mouse. Moreover, the inoculated mice showed no splenomegaly, which indicates that the mutants are highly attenuated. Importantly, the attenuation of in vitro and in vivo growth did not impair the ability of these mutants to confer long-term protective immunity in mice against challenge with B. abortus strain 2308. Vaccination of mice with either mutant induced humoral and cell-mediated immune responses, and provided significantly better protection than commercial B. abortus strain RB51 vaccine. These results suggest that highly attenuated BA15ΔcydCΔcydD and BA15ΔcydCΔpurD mutants can be used effectively as potential live vaccine candidates against bovine brucellosis.

  19. Radio-chemotherapy improves survival in IDH-mutant, 1p/19q non-codeleted secondary high-grade astrocytoma patients.

    PubMed

    Juratli, Tareq A; Lautenschläger, Tim; Geiger, Kathrin D; Pinzer, Thomas; Krause, Mechthild; Schackert, Gabriele; Krex, Dietmar

    2015-09-01

    Isocitrate dehydrogenase (IDH) mutations are beginning to drive decisions on therapy for glioma patients. Here we sought to determine the impact of adjuvant treatment in patients with IDH-mutant, 1p/19q non-codeleted secondary high-grade astrocytoma (sHGA) WHO grades III/IV. Clinical data of 109 sHGA patients grades III/IV, in addition to IDH mutation-, 1p/19q-codeletion- and MGMT-promoter methylation status-were retrospectively analyzed. Survival analysis in relation to adjuvant treatment modalities and molecular profiling were performed. Out of 109 patients, 88 patients (80.7 %) harbored IDH mutations, 30 patients had a 1p/19q-codeletion (27.5 %) and 69 patients (63.3 %) exhibited a methylated MGMT-promoter status. At a median follow-up of 9.8 years, 62 patients (57 %) died. The postsurgical treatment included: radio-chemotherapy (RT-CT; 54.5 %), RT alone (19.3 %), and CT alone (22.7 %). The median overall survival (OS) in the entire group was 3.4 years (1.9-6.7 years). Patients who received RT-CT had a significantly longer OS compared with those who underwent RT alone (6.5 vs. 1.2 years, HR 0.35, CI 0.32-0.51, p = 0.011). In the IDH-mutant 1p/19q non-codeleted sHGA subgroup the RT-CT cohort had a significantly longer OS in comparison to the RT cohort (6.4 vs. 1.2 years, HR 2.7, CI 1.1-6.5, p = 0.022). In the stepwise multivariable Cox model for OS of all 88 IDH-mutant sHGA patients, survival was strongly associated with only one factor, namely, adjuvant RT-CT at diagnosis of a sHGA. This retrospective long-term study demonstrates that RT and CT (mostly PCV) significantly improves progression-free and overall survival in IDH-mutant secondary high-grade astrocytoma patients, regardless of 1p/19q-codeletion status. PMID:26033545

  20. MnBr₂/18-crown-6 coordination complexes showing high room temperature luminescence and quantum yield.

    PubMed

    Hausmann, David; Kuzmanoski, Ana; Feldmann, Claus

    2016-04-21

    The reaction of manganese(ii) bromide and the crown ether 18-crown-6 in the ionic liquid [(n-Bu)3MeN][N(Tf)2] under mild conditions (80-130 °C) resulted in the formation of three different coordination compounds: MnBr2(18-crown-6) (), Mn3Br6(18-crown-6)2 () and Mn3Br6(18-crown-6) (). In general, the local coordination and the crystal structure of all compounds are driven by the mismatch between the small radius of the Mn(2+) cation (83 pm) and the ring opening of 18-crown-6 as a chelating ligand (about 300 pm). This improper situation leads to different types of coordination and bonding. MnBr2(18-crown-6) represents a molecular compound with Mn(2+) coordinated by two bromine atoms and only five oxygen atoms of 18-crown-6. Mn3Br6(18-crown-6)2 falls into a [MnBr(18-crown-6)](+) cation - with Mn(2+) coordinated by six oxygen atoms and Br - and a [MnBr(18-crown-6)MnBr4](-) anion. In this anion, Mn(2+) is coordinated by five oxygen atoms of the crown ether as well as by two bromine atoms, one of them bridging to an isolated (MnBr4) tetrahedron. Mn3Br6(18-crown-6), finally, forms an infinite, non-charged [Mn2(18-crown-6)(MnBr6)] chain. Herein, 18-crown-6 is exocyclically coordinated by two Mn(2+) cations. All compounds show intense luminescence in the yellow to red spectral range and exhibit remarkable quantum yields of 70% (Mn3Br6(18-crown-6)) and 98% (Mn3Br6(18-crown-6)2). The excellent quantum yield of Mn3Br6(18-crown-6)2 and its differentiation from MnBr2(18-crown-6) and Mn3Br6(18-crown-6) can be directly correlated to the local coordination. PMID:26956783

  1. Biosynthesis of high yield fatty acids from Chlorella vulgaris NIES-227 under nitrogen starvation stress during heterotrophic cultivation.

    PubMed

    Shen, Xiao-Fei; Chu, Fei-Fei; Lam, Paul K S; Zeng, Raymond J

    2015-09-15

    In this study the heterotrophic cultivation of Chlorella vulgaris NIES-227 fed with glucose was investigated systematically using six media types; combinations of nitrogen repletion/depletion and phosphorus repletion/limitation/depletion. It was found that a high yield of fatty acids (0.88 of fed glucose-COD) and a high content of fatty acid methyl esters (FAMEs) (89% of dry weight) were obtained under nitrogen starved conditions. To our knowledge it is the first report on such high COD conversion yield and FAME content in microalgae. The dominant fatty acid (>50%) was methyl oleate (C18:1), a desirable component for biodiesel synthesis. FAME content under nitrogen starved conditions was significantly higher than under nitrogen sufficient conditions, while phosphorus had no significant influence, indicating that nitrogen starvation was the real "fatty acids trigger" in heterotrophic cultivation. These findings could simplify the downstream extraction process, such as the extrusion of oil from soybeans, and could reduce operating costs by improving the fatty acid yield from waste COD.

  2. Prognostic significance of L1CAM expression and its association with mutant p53 expression in high-risk endometrial cancer.

    PubMed

    Van Gool, Inge C; Stelloo, Ellen; Nout, Remi A; Nijman, Hans W; Edmondson, Richard J; Church, David N; MacKay, Helen J; Leary, Alexandra; Powell, Melanie E; Mileshkin, Linda; Creutzberg, Carien L; Smit, Vincent T H B M; Bosse, Tjalling

    2016-02-01

    Studies in early-stage, predominantly low- and intermediate-risk endometrial cancer have demonstrated that L1 cell adhesion molecule (L1CAM) overexpression identifies patients at increased risk of recurrence, yet its prognostic significance in high-risk endometrial cancer is unclear. To evaluate this, its frequency, and the relationship of L1CAM with the established endometrial cancer biomarker p53, we analyzed the expression of both markers by immunohistochemistry in a pilot series of 116 endometrial cancers (86 endometrioid, 30 non-endometrioid subtype) with high-risk features (such as high tumor grade and deep myometrial invasion) and correlated results with clinical outcome. We used The Cancer Genome Atlas (TCGA) endometrial cancer series to validate our findings. Using the previously reported cutoff of 10% positive staining, 51/116 (44%) tumors were classified as L1CAM-positive, with no significant association between L1CAM positivity and the rate of distant metastasis (P=0.195). However, increasing the threshold for L1CAM positivity to 50% resulted in a reduction of the frequency of L1CAM-positive tumors to 24% (28/116), and a significant association with the rate of distant metastasis (P=0.018). L1CAM expression was strongly associated with mutant p53 in the high-risk and TCGA series (P<0.001), although a substantial fraction (36% of endometrioid, 10% of non-endometrioid morphology) of p53-mutant endometrial cancers displayed <10% L1CAM positivity. Moreover, 30% of p53-wild-type non-endometrioid endometrial cancers demonstrated diffuse L1CAM staining, suggesting p53-independent mechanisms of L1CAM overexpression. In conclusion, the previously proposed threshold for L1CAM positivity of >10% does not predict prognosis in high-risk endometrial cancer, whereas an alternative threshold (>50%) does. L1CAM expression is strongly, but not universally, associated with mutant p53, and may be strong enough for clinical implementation as prognostic marker in combination

  3. High-Throughput Cloning of Temperature-Sensitive Caenorhabditis elegans Mutants with Adult Syncytial Germline Membrane Architecture Defects

    PubMed Central

    Lowry, Josh; Yochem, John; Chuang, Chien-Hui; Sugioka, Kenji; Connolly, Amy A.; Bowerman, Bruce

    2015-01-01

    The adult Caenorhabditis elegans hermaphrodite gonad consists of two mirror-symmetric U-shaped arms, with germline nuclei located peripherally in the distal regions of each arm. The nuclei are housed within membrane cubicles that are open to the center, forming a syncytium with a shared cytoplasmic core called the rachis. As the distal germline nuclei progress through meiotic prophase, they move proximally and eventually cellularize as their compartments grow in size. The development and maintenance of this complex and dynamic germline membrane architecture are relatively unexplored, and we have used a forward genetic screen to identify 20 temperature-sensitive mutations in 19 essential genes that cause defects in the germline membrane architecture. Using a combined genome-wide SNP mapping and whole genome sequencing strategy, we have identified the causal mutations in 10 of these mutants. Four of the genes we have identified are conserved, with orthologs known to be involved in membrane biology, and are required for proper development or maintenance of the adult germline membrane architecture. This work provides a starting point for further investigation of the mechanisms that control the dynamics of syncytial membrane architecture during adult oogenesis. PMID:26311651

  4. Anistotropic yielding of rocks at high temperatures and pressures. Final report

    SciTech Connect

    Kronenberg, A.K.; Russell, J.E.; Carter, N.L.

    1990-10-14

    The anisotropic deformation of foliated and linealed rocks has been investigated, primarily to predict the mechanical response of rocks surrounding buried magma chambers to the stress fields generated by deep drilling. The principal application in this regard has been to evaluate, the scientific feasibility of extracting geothermal energy from buried magma chambers. Our approach has been to perform triaxial extension and compression tests at temperatures and pressures representative of the borehole environment on samples cored along six selected orientations and to fit the data to an orthohombric yield criterion. We have investigated Four-Mile gneiss (a strongly layered gneiss with well defined lineation), a biotite-rich schist, and Westerly granite (using a block oriented with respect to the granite`s rift, grain, and hardway). Progress has been made in three areas: the experimental determination of strength anisotropies for the three starting materials, theoretical treatment and modeling of the results, and characterization of fabrics surrounding magma bodies resulting from their diaperic emplacement into shallow portions of the Earth`s crust. In addition, results have been obtained for the tensile fracture of quartzite, basal slip and anisotropy of biotite single crystals, and anisotropic flow of bedded rocksalt.

  5. Template-particle stabilized bicontinuous emulsion yielding controlled assembly of hierarchical high-flux filtration membranes.

    PubMed

    Hess, Samuel C; Kohll, A Xavier; Raso, Renzo A; Schumacher, Christoph M; Grass, Robert N; Stark, Wendelin J

    2015-01-14

    A novel solvent-evaporation-based process that exploits template-particle stabilized bicontinuous emulsions for the formation of previously unreached membrane morphologies is reported in this article. Porous membranes have a wide range of applications spanning from water filtration, pharmaceutical purification, and battery separators to scaffolds for tissue engineering. Different situations require different membrane morphologies including various pore sizes and pore gradients. However, most of the previously reported membrane preparation procedures are restricted to specific morphologies and morphology alterations require an extensive optimization process. The tertiary system presented in this article, which consists of a poly(ether sulfone)/dimethylacetamide (PES/DMAc) solution, glycerol, and ZnO-nanoparticles, allows simple and exact tuning of pore diameters ranging from sub-20 nm, up to 100 nm. At the same time, the pore size gradient is controlled from 0 up to 840%/μm yielding extreme asymmetry. In addition to structural analysis, water flux rates of over 5600 L m(-2) h(-1) are measured for membranes retaining 45 nm silica beads.

  6. Template-particle stabilized bicontinuous emulsion yielding controlled assembly of hierarchical high-flux filtration membranes.

    PubMed

    Hess, Samuel C; Kohll, A Xavier; Raso, Renzo A; Schumacher, Christoph M; Grass, Robert N; Stark, Wendelin J

    2015-01-14

    A novel solvent-evaporation-based process that exploits template-particle stabilized bicontinuous emulsions for the formation of previously unreached membrane morphologies is reported in this article. Porous membranes have a wide range of applications spanning from water filtration, pharmaceutical purification, and battery separators to scaffolds for tissue engineering. Different situations require different membrane morphologies including various pore sizes and pore gradients. However, most of the previously reported membrane preparation procedures are restricted to specific morphologies and morphology alterations require an extensive optimization process. The tertiary system presented in this article, which consists of a poly(ether sulfone)/dimethylacetamide (PES/DMAc) solution, glycerol, and ZnO-nanoparticles, allows simple and exact tuning of pore diameters ranging from sub-20 nm, up to 100 nm. At the same time, the pore size gradient is controlled from 0 up to 840%/μm yielding extreme asymmetry. In addition to structural analysis, water flux rates of over 5600 L m(-2) h(-1) are measured for membranes retaining 45 nm silica beads. PMID:25513883

  7. Discrete Electronic Bands in Semiconductors and Insulators: Potential High-Light-Yield Scintillators

    DOE PAGESBeta

    Shi, Hongliang; Du, Mao-Hua

    2015-05-12

    Bulk semiconductors and insulators typically have continuous valence and conduction bands. In this paper, we show that valence and conduction bands of a multinary semiconductor or insulator can be split to narrow discrete bands separated by large energy gaps. This unique electronic structure is demonstrated by first-principles calculations in several quaternary elpasolite compounds, i.e., Cs2NaInBr6, Cs2NaBiCl6, and Tl2NaBiCl6. The narrow discrete band structure in these quaternary elpasolites is due to the large electronegativity difference among cations and the large nearest-neighbor distances in cation sublattices. We further use Cs2NaInBr6 as an example to show that the narrow bands can stabilize self-trappedmore » and dopant-bound excitons (in which both the electron and the hole are strongly localized in static positions on adjacent sites) and promote strong exciton emission at room temperature. The discrete band structure should further suppress thermalization of hot carriers and may lead to enhanced impact ionization, which is usually considered inefficient in bulk semiconductors and insulators. Finally, these characteristics can enable efficient room-temperature light emission in low-gap scintillators and may overcome the light-yield bottleneck in current scintillator research.« less

  8. Discrete Electronic Bands in Semiconductors and Insulators: Potential High-Light-Yield Scintillators

    SciTech Connect

    Shi, Hongliang; Du, Mao-Hua

    2015-05-12

    Bulk semiconductors and insulators typically have continuous valence and conduction bands. In this paper, we show that valence and conduction bands of a multinary semiconductor or insulator can be split to narrow discrete bands separated by large energy gaps. This unique electronic structure is demonstrated by first-principles calculations in several quaternary elpasolite compounds, i.e., Cs2NaInBr6, Cs2NaBiCl6, and Tl2NaBiCl6. The narrow discrete band structure in these quaternary elpasolites is due to the large electronegativity difference among cations and the large nearest-neighbor distances in cation sublattices. We further use Cs2NaInBr6 as an example to show that the narrow bands can stabilize self-trapped and dopant-bound excitons (in which both the electron and the hole are strongly localized in static positions on adjacent sites) and promote strong exciton emission at room temperature. The discrete band structure should further suppress thermalization of hot carriers and may lead to enhanced impact ionization, which is usually considered inefficient in bulk semiconductors and insulators. Finally, these characteristics can enable efficient room-temperature light emission in low-gap scintillators and may overcome the light-yield bottleneck in current scintillator research.

  9. High-yield production of graphene by liquid-phase exfoliation of graphite.

    PubMed

    Hernandez, Yenny; Nicolosi, Valeria; Lotya, Mustafa; Blighe, Fiona M; Sun, Zhenyu; De, Sukanta; McGovern, I T; Holland, Brendan; Byrne, Michele; Gun'Ko, Yurii K; Boland, John J; Niraj, Peter; Duesberg, Georg; Krishnamurthy, Satheesh; Goodhue, Robbie; Hutchison, John; Scardaci, Vittorio; Ferrari, Andrea C; Coleman, Jonathan N

    2008-09-01

    Fully exploiting the properties of graphene will require a method for the mass production of this remarkable material. Two main routes are possible: large-scale growth or large-scale exfoliation. Here, we demonstrate graphene dispersions with concentrations up to approximately 0.01 mg ml(-1), produced by dispersion and exfoliation of graphite in organic solvents such as N-methyl-pyrrolidone. This is possible because the energy required to exfoliate graphene is balanced by the solvent-graphene interaction for solvents whose surface energies match that of graphene. We confirm the presence of individual graphene sheets by Raman spectroscopy, transmission electron microscopy and electron diffraction. Our method results in a monolayer yield of approximately 1 wt%, which could potentially be improved to 7-12 wt% with further processing. The absence of defects or oxides is confirmed by X-ray photoelectron, infrared and Raman spectroscopies. We are able to produce semi-transparent conducting films and conducting composites. Solution processing of graphene opens up a range of potential large-area applications, from device and sensor fabrication to liquid-phase chemistry.

  10. Experimental Determination of DT Yield in High Current DD Dense Plasma Focii

    SciTech Connect

    Lowe, D. R.; Hagen, E. C.; Meehan, B. T.; Springs, R. K.; O'Brien, R. J.

    2013-06-18

    Dense Plasma Focii (DPF), which utilize deuterium gas to produce 2.45 MeV neutrons, may in fact also produce DT fusion neutrons at 14.1 MeV due to the triton production in the DD reaction. If beam-target fusion is the primary producer of fusion neutrons in DPFs, it is possible that ejected tritons from the first pinch will interact with the second pinch, and so forth. The 2 MJ DPF at National Security Technologies’ Losee Road Facility is able to, and has produced, over 1E12 DD neutrons per pulse, allowing an accurate measurement of the DT/DD ratio. The DT/DD ratio was experimentally verified by using the (n,2n) reaction in a large piece of praseodymium metal, which has a threshold reaction of 8 MeV, and is widely used as a DT yield measurement system1. The DT/DD ratio was experimentally determined for over 100 shots, and then compared to independent variables such as tube pressure, number of pinches per shot, total current, pinch current and charge voltage.

  11. Food webs: ordering species according to body size yields high degree of intervality.

    PubMed

    Zook, Alexander E; Eklof, Anna; Jacob, Ute; Allesina, Stefano

    2011-02-21

    Food webs, the networks describing "who eats whom" in an ecosystem, are nearly interval, i.e. there is a way to order the species so that almost all the resources of each consumer are adjacent in the ordering. This feature has important consequences, as it means that the structure of food webs can be described using a single (or few) species' traits. Moreover, exploiting the quasi-intervality found in empirical webs can help build better models for food web structure. Here we investigate which species trait is a good proxy for ordering the species to produce quasi-interval orderings. We find that body size produces a significant degree of intervality in almost all food webs analyzed, although it does not match the maximum intervality for the networks. There is also a great variability between webs. Other orderings based on trophic levels produce a lower level of intervality. Finally, we extend the concept of intervality from predator-centered (in which resources are in intervals) to prey-centered (in which consumers are in intervals). In this case as well we find that body size yields a significant, but not maximal, level of intervality. These results show that body size is an important, although not perfect, trait that shapes species interactions in food webs. This has important implications for the formulation of simple models used to construct realistic representations of food webs.

  12. Randomized trial of oral cyclophosphamide and veliparib in high-grade serous ovarian, primary peritoneal, or fallopian tube cancers, or BRCA-mutant ovarian cancer

    PubMed Central

    Kummar, Shivaani; Oza, Amit M.; Fleming, Gini F.; Sullivan, Daniel M.; Gandara, David R.; Naughton, Michael J.; Villalona-Calero, Miguel A.; Morgan, Robert J.; Szabo, Peter M.; Youn, Ahrim; Chen, Alice P.; Ji, Jiuping; Allen, Deborah E.; Lih, Chih-Jian; Mehaffey, Michele G.; Walsh, William D.; McGregor, Paul M.; Steinberg, Seth M.; Williams, Paul M.; Kinders, Robert J.; Conley, Barbara A.; Simon, Richard M.; Doroshow, James H.

    2015-01-01

    Purpose Veliparib, a poly(ADP-ribose) polymerase (PARP) inhibitor, demonstrated clinical activity in combination with oral cyclophosphamide in patients with BRCA-mutant solid tumors in a phase 1 trial. To define the relative contribution of PARP inhibition to the observed clinical activity, we conducted a randomized phase 2 trial to determine the response rate of veliparib in combination with cyclophosphamide compared to cyclophosphamide alone in patients with pretreated BRCA-mutant ovarian cancer or in patients with pretreated primary peritoneal, fallopian tube, or high-grade serous ovarian cancers (HGSOC). Methods Adult patients were randomized to receive cyclophosphamide alone (50 mg orally once daily) or with veliparib (60 mg orally once daily) in 21-day cycles. Crossover to the combination was allowed at disease progression. Results Seventy-five patients were enrolled and 72 were evaluable for response; 38 received cyclophosphamide alone and 37 the combination as their initial treatment regimen. Treatment was well tolerated. One complete response was observed in each arm, with three partial responses (PR) in the combination arm and six PRs in the cyclophosphamide alone arm. Genetic sequence and expression analyses were performed for 211 genes involved in DNA repair; none of the detected genetic alterations were significantly associated with treatment benefit. Conclusion This is the first trial that evaluated single agent, low dose cyclophosphamide in HGSOC, peritoneal, fallopian tube, and BRCA-mutant ovarian cancers. It was well tolerated and clinical activity was observed; the addition of veliparib at 60 mg daily did not improve either the response rate or the median progression free survival. PMID:25589624

  13. Novel mutant human immunodeficiency virus type 1 strains with high degree of resistance to cynomolgus macaque TRIMCyp generated by random mutagenesis.

    PubMed

    Sultana, Tahmina; Nakayama, Emi E; Tobita, Satoshi; Yokoyama, Masaru; Seki, Yohei; Saito, Akatsuki; Nomaguchi, Masako; Adachi, Akio; Akari, Hirofumi; Sato, Hironori; Shioda, Tatsuo

    2016-04-01

    Old World monkey TRIM5α strongly suppresses human immunodeficiency virus type 1 (HIV-1) replication. A fusion protein comprising cynomolgus macaque (CM) TRIM5 and cyclophilin A (CM TRIMCyp) also potently suppresses HIV-1 replication. However, CM TRIMCyp fails to suppress a mutant HIV-1 that encodes a mutant capsid protein containing a SIVmac239-derived loop between α-helices 4 and 5 (L4/5). There are seven amino acid differences between L4/5 of HIV-1 and SIVmac239. Here, we investigated the minimum numbers of amino acid substitutions that would allow HIV-1 to evade CM TRIMCyp-mediated suppression. We performed random PCR mutagenesis to construct a library of HIV-1 variants containing mutations in L4/5, and then we recovered replication-competent viruses from CD4+ MT4 cells that expressed high levels of CM TRIMCyp. CM TRIMCyp-resistant viruses were obtained after three rounds of selection in MT4 cells expressing CM TRIMCyp and these were found to contain four amino acid substitutions (H87R, A88G, P90D and P93A) in L4/5. We then confirmed that these substitutions were sufficient to confer CM TRIMCyp resistance to HIV-1. In a separate experiment using a similar method, we obtained novel CM TRIM5α-resistant HIV-1 strains after six rounds of selection and rescue. Analysis of these mutants revealed that V86A and G116E mutations in the capsid region conferred partial resistance to CM TRIM5α without substantial fitness cost when propagated in MT4 cells expressing CM TRIM5α. These results confirmed and further extended the previous notion that CM TRIMCyp and CM TRIM5α recognize the HIV-1 capsid in different manners. PMID:26795727

  14. The Effect of Body Energy Reserve Mobilization on the Fatty Acid Profile of Milk in High-yielding Cows

    PubMed Central

    Nogalski, Zenon; Wroński, Marek; Sobczuk-Szul, Monika; Mochol, Magdalena; Pogorzelska, Paulina

    2012-01-01

    We investigated the effect of the amount of body condition loss in the dry period and early lactation in 42 high-yielding Holstein-Friesian cows on milk yield and the share of fatty acids in milk fat. Energy reserves were estimated based on the body condition scoring (BCS) and backfat thickness (BFT). Milk yield and milk composition were determined over 305-d lactation. From d 6 to 60 of lactation, the concentrations of 43 fatty acids in milk fat were determined by gas chromatography. Cows were categorized based on body condition loss from the beginning of the dry period to the lowest point of the BCS curve in early lactation into three groups: low condition loss group (L) ≤0.5 points (n = 14); moderate condition loss group (M) 0.75 to 1.0 points (n = 16) and high condition loss group (H) >1.0 points (n = 12). Cows whose body energy reserves were mobilized at 0.8 BCS and 11 mm BFT, produced 12,987 kg ECM over 305-d lactation, i.e. 1,429 kg ECM more than cows whose BCS and BFT decreased by 0.3 and 5 mm, respectively. In group H, milk yield reached 12,818 kg ECM at body fat reserve mobilization of 1.3 BCS and 17 mm BFT. High mobilization of body fat reserves led to a significant (approx. 5%) increase in the concentrations of monounsaturated fatty acids-MUFA (mostly C18:1 cis-9, followed by C18:1 trans-11), a significant decrease in the levels of fatty acids adversely affecting human health, and a drop in the content of linoleic acid, arachidonic acid and docosahexaenoic acid in milk fat. In successive weeks of lactation, an improved energy balance contributed to a decrease in the concentrations of unsaturated fatty acids (UFA) and an increase in the conjugated linoleic acid (CLA) content of milk fat. PMID:25049536

  15. Embedded pitch adapters: A high-yield interconnection solution for strip sensors

    NASA Astrophysics Data System (ADS)

    Ullán, M.; Allport, P. P.; Baca, M.; Broughton, J.; Chisholm, A.; Nikolopoulos, K.; Pyatt, S.; Thomas, J. P.; Wilson, J. A.; Kierstead, J.; Kuczewski, P.; Lynn, D.; Hommels, L. B. A.; Fleta, C.; Fernandez-Tejero, J.; Quirion, D.; Bloch, I.; Díez, S.; Gregor, I. M.; Lohwasser, K.; Poley, L.; Tackmann, K.; Hauser, M.; Jakobs, K.; Kuehn, S.; Mahboubi, K.; Mori, R.; Parzefall, U.; Clark, A.; Ferrere, D.; Gonzalez Sevilla, S.; Ashby, J.; Blue, A.; Bates, R.; Buttar, C.; Doherty, F.; McMullen, T.; McEwan, F.; O'Shea, V.; Kamada, S.; Yamamura, K.; Ikegami, Y.; Nakamura, K.; Takubo, Y.; Unno, Y.; Takashima, R.; Chilingarov, A.; Fox, H.; Affolder, A. A.; Casse, G.; Dervan, P.; Forshaw, D.; Greenall, A.; Wonsak, S.; Wormald, M.; Cindro, V.; Kramberger, G.; Mandić, I.; Mikuž, M.; Gorelov, I.; Hoeferkamp, M.; Palni, P.; Seidel, S.; Taylor, A.; Toms, K.; Wang, R.; Hessey, N. P.; Valencic, N.; Hanagaki, K.; Dolezal, Z.; Kodys, P.; Bohm, J.; Mikestikova, M.; Bevan, A.; Beck, G.; Milke, C.; Domingo, M.; Fadeyev, V.; Galloway, Z.; Hibbard-Lubow, D.; Liang, Z.; Sadrozinski, H. F.-W.; Seiden, A.; To, K.; French, R.; Hodgson, P.; Marin-Reyes, H.; Parker, K.; Jinnouchi, O.; Hara, K.; Bernabeu, J.; Civera, J. V.; Garcia, C.; Lacasta, C.; Marti i Garcia, S.; Rodriguez, D.; Santoyo, D.; Solaz, C.; Soldevila, U.

    2016-09-01

    A proposal to fabricate large area strip sensors with integrated, or embedded, pitch adapters is presented for the End-cap part of the Inner Tracker in the ATLAS experiment. To implement the embedded pitch adapters, a second metal layer is used in the sensor fabrication, for signal routing to the ASICs. Sensors with different embedded pitch adapters have been fabricated in order to optimize the design and technology. Inter-strip capacitance, noise, pick-up, cross-talk, signal efficiency, and fabrication yield have been taken into account in their design and fabrication. Inter-strip capacitance tests taking into account all channel neighbors reveal the important differences between the various designs considered. These tests have been correlated with noise figures obtained in full assembled modules, showing that the tests performed on the bare sensors are a valid tool to estimate the final noise in the full module. The full modules have been subjected to test beam experiments in order to evaluate the incidence of cross-talk, pick-up, and signal loss. The detailed analysis shows no indication of cross-talk or pick-up as no additional hits can be observed in any channel not being hit by the beam above 170 mV threshold, and the signal in those channels is always below 1% of the signal recorded in the channel being hit, above 100 mV threshold. First results on irradiated mini-sensors with embedded pitch adapters do not show any change in the interstrip capacitance measurements with only the first neighbors connected.

  16. Taurus Littrow Pyroclastic Deposit: High-Yield Feedstock for Lunar Oxygen

    NASA Technical Reports Server (NTRS)

    Allen, Carlton C.

    2015-01-01

    Future human habitation of the Moon will likely require the use of locally derived materials because of the high cost of transportation from Earth. Oxygen, extracted from oxides and silicates, is a potentially abundant lunar resource vital for life support and spacecraft propulsion. The anticipated costs of supplying all oxygen needs for a lunar base from Earth are high enough to warrant serious study of oxygen production from local resources.

  17. Effects of Fluctuating Environments on the Selection of High Yielding Microalgae

    SciTech Connect

    Benemann, J. R.; Tillett, D. M.

    1987-02-27

    Microalgae have the potential of producing biomass with a high content of lipids at high productivities using seawater or saline ground water resources. Microalgal lipids are similar to vegetable oils and suitable for processing to liquid fuels. Engineering cost analysis studies have concluded that, at a favorable site, microalgae cultivation for fuel production could be economically viable. The major uncertainties involve the microalgae themselves: biomass and lipid productivity and culture stability.

  18. High yield production of sugars from deproteinated palm kernel cake under microwave irradiation via dilute sulfuric acid hydrolysis.

    PubMed

    Fan, Suet-Pin; Jiang, Li-Qun; Chia, Chin-Hua; Fang, Zhen; Zakaria, Sarani; Chee, Kah-Leong

    2014-02-01

    Recent years, great interest has been devoted to the conversion of biomass-derived carbohydrate into sugars, such as glucose, mannose and fructose. These are important versatile intermediate products that are easily processed into high value-added biofuels. In this work, microwave-assisted dilute sulfuric acid hydrolysis of deproteinated palm kernel cake (DPKC) was systematically studied using Response Surface Methodology. The highest mannose yield (92.11%) was achieved at the optimized condition of 148°C, 0.75N H2SO4, 10min 31s and substrate to solvent (SS) ratio (w/v) of 1:49.69. Besides that, total fermentable sugars yield (77.11%), was obtained at 170°C, 0.181N H2SO4, 6min 6s and SS ratio (w/v) of 1:40. Ridge analysis was employed to further verify the optimum conditions. Thus, this work provides fundamental data of the practical use of DPKC as low cost, high yield and environmental-friendly material for the production of mannose and other sugars.

  19. Mutant pfcrt "SVMNT" haplotype and wild type pfmdr1 "N86" are endemic in Plasmodium vivax dominated areas of India under high chloroquine exposure

    PubMed Central

    2012-01-01

    Background Chloroquine resistance (CQR) phenotype in Plasmodium falciparum is associated with mutations in pfcrt and pfmdr-1 genes. Mutations at amino acid position 72-76 of pfcrt gene, here defined as pfcrt haplotype are associated with the geographic origin of chloroquine resistant parasite. Here, mutations at 72-76 and codon 220 of pfcrt gene and N86Y pfmdr-1 mutation were studied in blood samples collected across 11 field sites, inclusive of high and low P. falciparum prevalent areas in India. Any probable correlation between these mutations and clinical outcome of CQ treatment was also investigated. Methods Finger pricked blood spotted on Whatman No.3 papers were collected from falciparum malaria patients of high and low P. falciparum prevalent areas. For pfcrt haplotype investigation, the parasite DNA was extracted from blood samples and used for PCR amplification, followed by partial sequencing of the pfcrt gene. For pfmdr-1 N86Y mutation, the PCR product was subjected to restriction digestion with AflIII endonuclease enzyme. Results In 240 P. falciparum isolates with reported in vivo CQ therapeutic efficacy, the analysis of mutations in pfcrt gene shows that mutant SVMNT-S (67.50%) and CVIET-S (23.75%) occurred irrespective of clinical outcome and wild type CVMNK-A (7.91%) occurred only in adequate clinical and parasitological response samples. Of 287 P. falciparum isolates, SVMNTS 192 (66.89%) prevailed in all study sites and showed almost monomorphic existence (98.42% isolates) in low P. falciparum prevalent areas. However, CVIETS-S (19.51%) and CVMNK-A (11.84%) occurrence was limited to high P. falciparum prevalent areas. Investigation of pfmdr-1 N86Y mutation shows no correlation with clinical outcomes. The wild type N86 was prevalent in all the low P. falciparum prevalent areas (94.48%). However, mutant N86Y was comparably higher in numbers at the high P. falciparum prevalent areas (42.76%). Conclusions The wild type pfcrt gene is linked to chloroquine

  20. Effects of high-temperature isochoric pre-treatment on the methane yields of cattle, pig and chicken manure.

    PubMed

    Raju, Chitra Sangaraju; Sutaryo, Sutaryo; Ward, Alastair James; Møller, Henrik Bjarne

    2013-01-01

    Cattle manure, dewatered pig manure and chicken manure were pre-treated in a high-temperature reactor under isochoric conditions for 15 min at temperatures between 100 and 225 degrees C with 25 degrees C intervals to study the effect on their methane yield. After 27 days of batch incubation, cattle manure showed a significant improvement in its biochemical methane potential (BMP) of 13% at 175 degrees C and 21% at 200 degrees C. Pig manure showed improvements at temperatures of 125 degrees C and above, with a maximum 29% increase in yield at 200 degrees C. The BMP of chicken manure was reduced by 18% at 225 degrees C, but at lower temperatures there were no significant changes. It was found that this method of pre-treatment could be feasible if sufficient surplus energy was available or if the energy used in the pre-treatment could be recovered.

  1. Neutron Yield and Ion Temperature from DD and DT Fusion in National Ignition Facility High-Foot Implosions

    NASA Astrophysics Data System (ADS)

    Knauer, J. P.; Gatu Johnson, M.; Frenje, J. A.; Petrasso, R. D.; Caggiano, J. A.; Callahan, D. A.; Casey, D. T.; Cerjan, C. J.; Doeppner, T.; Eckart, M. J.; Grim, G. P.; Hartouni, E. P.; Hatarik, R.; Hinkel, D. E.; Hurricane, O. A.; Kritcher, A.; Le Pape, S.; Ma, T.; Munro, D. H.; Patel, P. K.; Ralph, J. E.; Sayre, D. B.; Spears, B. K.; Yeamans, C. B.; Kilkenny, J. D.

    2015-11-01

    Simultaneous measures of neutrons emitted from DT fusion implosions are postulated to provide insight into the fuel conditions during neutron emission. Neutron spectral diagnostics of National Ignition Facility ``high-foot'' implosions measure both the DT and DD fusion neutron spectra. Equivalent ion temperature is measured from the width of the DT and DD neutron emission and the respective yields from the peak areas. This work has focused on reasons for differing inferred temperatures from the DT and DD spectra and the yield ratio. Spatial and temporal averages of the DT and DD reactivities as corrections to the homogeneous and static temperature distributions are shown. This material is based upon work supported by the Department of Energy National Nuclear Security Administration under Award Number DE-NA0001944.

  2. Structure-Triggered High Quantum Yield Luminescence and Switchable Dielectric Properties in Manganese(II) Based Hybrid Compounds.

    PubMed

    Wang, Zhong-Xia; Li, Peng-Fei; Liao, Wei-Qiang; Tang, Yuanyuan; Ye, Heng-Yun; Zhang, Yi

    2016-04-01

    Two new manganese(II) based organic-inorganic hybrid compounds, C11H21Cl3MnN2 (1) and C11H22Cl4MnN2 (2), with prominent photoluminescence and dielectric properties were synthesized by solvent modulation. Compound 1 with novel trigonal bipyramidal geometry exhibits bright red luminescence with a lifetime of 2.47 ms and high quantum yield of 35.8 %. Compound 2 with tetrahedral geometry displays intense long-lived (1.54 ms) green light emission with higher quantum yield of 92.3 %, accompanied by reversible solid-state phase transition at 170 K and a distinct switchable dielectric property. The better performance of 2 results from the structure, including a discrete organic cation moiety and inorganic metal anion framework, which gives the cations large freedom of motion.

  3. High yield synthesis of Ni-BTC metal-organic framework with ultrasonic irradiation: Role of polar aprotic DMF solvent.

    PubMed

    Israr, Farrukh; Chun, Daye; Kim, Yeongmin; Kim, Duk Kyung

    2016-07-01

    Nickel based porous solid was synthesized with 20 kHz ultrasonic irradiation. The reaction of Ni(II) nitrate hexahydrate with 1,3,5-benzene tricarboxylic acid in N,N-Dimethylformamide (DMF) as the sole solvent under ultrasonic radiation produced porous Ni-BTC MOF. Choice of correct solvent for the ultrasonic treatment was proven important. The effect of varying ultrasonic powers (40%, 60% and 80% of 750 W) along with different temperature conditions (50 °C, 60 °C, 70 °C and 80 °C) influenced the respective yield. A very high yield of 88% Ni-BTC MOF was obtained from 80% ultrasonic power at 60 °C. BET surface areas of the MOF crystals measured by N2 gas adsorption isotherms were in the range of 960-1000 m(2)/g.

  4. Yield and temporal characterization of high-order harmonics from intense midinfrared excitation of a cesium vapor

    SciTech Connect

    Clatterbuck, T.O.; Lyngaa, C.; Paul, P.M.; DiMauro, L.F.; Gaarde, M.B.; Schafer, K.J.; Agostini, P.; Kulander, K.C.; Walmsley, I.

    2004-03-01

    Cesium vapor interacting with a tightly focused, intense midinfrared (3.4 {mu}m) source produces high harmonic radiation in the visible/UV spectral range. The measured yields of harmonic orders 9-17 are found to yield good quantitative agreement with numerical simulations that include both the single-atom and the macroscopic response. The 5th-9th harmonic orders are generated with sufficient pulse energies ({approx}100 pJ) for direct temporal measurements using an autocorrelation method and when correlated with bandwidth measurements are found not to be transform limited. A blueshift connected to a strong time-dependent ionization appears to be the cause of this spectral broadening.

  5. Yield and temporal characterization of high-order harmonics from intense midinfrared excitation of a cesium vapor

    NASA Astrophysics Data System (ADS)

    Clatterbuck, T. O.; Lyngå, C.; Paul, P. M.; Dimauro, L. F.; Gaarde, M. B.; Schafer, K. J.; Agostini, P.; Kulander, K. C.; Walmsley, I.

    2004-03-01

    Cesium vapor interacting with a tightly focused, intense midinfrared (3.4 μm) source produces high harmonic radiation in the visible/UV spectral range. The measured yields of harmonic orders 9 17 are found to yield good quantitative agreement with numerical simulations that include both the single-atom and the macroscopic response. The 5th 9th harmonic orders are generated with sufficient pulse energies (˜100 pJ) for direct temporal measurements using an autocorrelation method and when correlated with bandwidth measurements are found not to be transform limited. A blueshift connected to a strong time-dependent ionization appears to be the cause of this spectral broadening.

  6. Physiological Mechanisms Underlying the High-Grain Yield and High-Nitrogen Use Efficiency of Elite Rice Varieties under a Low Rate of Nitrogen Application in China.

    PubMed

    Wu, Lilian; Yuan, Shen; Huang, Liying; Sun, Fan; Zhu, Guanglong; Li, Guohui; Fahad, Shah; Peng, Shaobing; Wang, Fei

    2016-01-01

    Selecting rice varieties with a high nitrogen (N) use efficiency (NUE) is the best approach to reduce N fertilizer application in rice production and is one of the objectives of the Green Super Rice (GSR) Project in China. However, the performance of elite candidate GSR varieties under low N supply remains unclear. In the present study, differences in the grain yield and NUE of 13 and 14 candidate varieties with two controls were determined at a N rate of 100 kg ha(-1) in field experiments in 2014 and 2015, respectively. The grain yield for all of the rice varieties ranged from 8.67 to 11.09 t ha(-1), except for a japonica rice variety YG29, which had a grain yield of 6.42 t ha(-1). HY549 and YY4949 produced the highest grain yield, reflecting a higher biomass production and harvest index in 2014 and 2015, respectively. Total N uptake at maturity (TNPM) ranged from 144 to 210 kg ha(-1), while the nitrogen use efficiency for grain production (NUEg) ranged from 35.2 to 62.0 kg kg(-1). Both TNPM and NUEg showed a significant quadratic correlation with grain yield, indicating that it is possible to obtain high grain yield and NUEg with the reduction of TNPM. The correlation between N-related parameters and yield-related traits suggests that promoting pre-heading growth could increase TNPM, while high biomass accumulation during the grain filling period and large panicles are important for a higher NUEg. In addition, there were significant and negative correlations between the NUEg and N concentrations in leaf, stem, and grain tissues at maturity. Further improvements in NUEg require a reduction in the stem N concentration but not the leaf N concentration. The daily grain yield was the only parameter that significantly and positively correlated with both TNPMand NUEg. This study determined variations in the grain yield and NUE of elite candidate GSR rice varieties and provided plant traits that could be used as selection criteria in breeding N-efficient rice varieties

  7. Physiological Mechanisms Underlying the High-Grain Yield and High-Nitrogen Use Efficiency of Elite Rice Varieties under a Low Rate of Nitrogen Application in China

    PubMed Central

    Wu, Lilian; Yuan, Shen; Huang, Liying; Sun, Fan; Zhu, Guanglong; Li, Guohui; Fahad, Shah; Peng, Shaobing; Wang, Fei

    2016-01-01

    Selecting rice varieties with a high nitrogen (N) use efficiency (NUE) is the best approach to reduce N fertilizer application in rice production and is one of the objectives of the Green Super Rice (GSR) Project in China. However, the performance of elite candidate GSR varieties under low N supply remains unclear. In the present study, differences in the grain yield and NUE of 13 and 14 candidate varieties with two controls were determined at a N rate of 100 kg ha−1 in field experiments in 2014 and 2015, respectively. The grain yield for all of the rice varieties ranged from 8.67 to 11.09 t ha−1, except for a japonica rice variety YG29, which had a grain yield of 6.42 t ha−1. HY549 and YY4949 produced the highest grain yield, reflecting a higher biomass production and harvest index in 2014 and 2015, respectively. Total N uptake at maturity (TNPM) ranged from 144 to 210 kg ha−1, while the nitrogen use efficiency for grain production (NUEg) ranged from 35.2 to 62.0 kg kg−1. Both TNPM and NUEg showed a significant quadratic correlation with grain yield, indicating that it is possible to obtain high grain yield and NUEg with the reduction of TNPM. The correlation between N-related parameters and yield-related traits suggests that promoting pre-heading growth could increase TNPM, while high biomass accumulation during the grain filling period and large panicles are important for a higher NUEg. In addition, there were significant and negative correlations between the NUEg and N concentrations in leaf, stem, and grain tissues at maturity. Further improvements in NUEg require a reduction in the stem N concentration but not the leaf N concentration. The daily grain yield was the only parameter that significantly and positively correlated with both TNPMand NUEg. This study determined variations in the grain yield and NUE of elite candidate GSR rice varieties and provided plant traits that could be used as selection criteria in breeding N-efficient rice varieties

  8. De novo Comparative Transcriptome Analysis of Acremonium chrysogenum: High-Yield and Wild-Type Strains of Cephalosporin C Producer

    PubMed Central

    Liu, Yan; Xie, Liping; Gong, Guihua; Zhang, Wei; Zhu, Baoquan; Hu, Youjia

    2014-01-01

    β-lactam antibiotics are widely used in clinic. Filamentous fungus Acremonium chrysogenum is an important industrial fungus for the production of CPC, one of the major precursors of β-lactam antibiotics. Although its fermentation yield has been bred significantly over the past decades, little is known regarding molecular changes between the industrial strain and the wild type strain. This limits the possibility to improve CPC production further by molecular breeding. Comparative transcriptome is a powerful tool to understand the molecular mechanisms of CPC industrial high yield producer compared to wild type. A total of 57 million clean sequencing reads with an average length of 100 bp were generated from Illumina sequencing platform. 22,878 sequences were assembled. Among the assembled unigenes, 9502 were annotated and 1989 annotated sequences were assigned to 121 pathways by searching against the Kyoto Encyclopedia of Genes and Genomes pathway (KEGG) database. Furthermore, we compared the transcriptome differences between a high-yield and a wild-type strain during fermentation. A total of 4329 unigenes with significantly different transcription level were identified, among which 1737 were up-regulated and 2592 were down-regulated. 24 pathways were subsequently determined which involve glycerolipid metabolism, galactose metabolism, and pyrimidine metabolism. We also examined the transcription levels of 18 identified genes, including 11 up-regulated genes and 7 down-regulated genes using reverse transcription quantitative -PCR (RT-qPCR). The results of RT-qPCR were consistent with the Illumina sequencing. In this study, the Illumina sequencing provides the most comprehensive sequences for gene expression profile of Acremonium chrysogenum and allows de novo transcriptome assembly while lacking genome information. Comparative analysis of RNA-seq data reveals the complexity of the transcriptome in the fermentation of different yield strains. This is an important

  9. High-Yield Endoglucanase Production by Trichoderma harzianum IOC-3844 Cultivated in Pretreated Sugarcane Mill Byproduct

    PubMed Central

    de Castro, Aline Machado; Ferreira, Marcela Costa; da Cruz, Juliana Cunha; Pedro, Kelly Cristina Nascimento Rodrigues; Carvalho, Daniele Fernandes; Leite, Selma Gomes Ferreira; Pereira, Nei

    2010-01-01

    The low-cost production of cellulolytic complexes presenting high action at mild conditions and well-balanced cellulase activities is one of the major bottlenecks for the economical viability of the production of cellulosic ethanol. In the present paper, the filamentous fungus Trichoderma harzianum IOC-3844 was used for the production of cellulases from a pretreated sugarcane bagasse (namely, cellulignin), by submerged fermentation. This fungal strain produced high contents of endoglucanase activity (6,358 U·L−1) after 72 hours of process, and further relevant β-glucosidase and FPase activities (742 and 445 U·L−1, resp.). The crude enzyme extract demonstrated appropriate characteristics for its application in cellulose hydrolysis, such as high thermal stability at up to 50°C, accessory xylanase activity, and absence of proteolytic activity towards azocasein. This strain showed, therefore, potential for the production of complete cellulolytic complexes aiming at the saccharification of lignocellulosic materials. PMID:21048871

  10. Physicochemical origin of high correlation between thermal stability of a protein and its packing efficiency: a theoretical study for staphylococcal nuclease mutants

    PubMed Central

    Oda, Koji; Kinoshita, Masahiro

    2015-01-01

    There is an empirical rule that the thermal stability of a protein is related to the packing efficiency or core volume of the folded state and the protein tends to exhibit higher stability as the backbone and side chains are more closely packed. Previously, the wild type and its nine mutants of staphylococcal nuclease were compared by examining their folded structures. The results obtained were as follows: The stability was not correlated with the number of intramolecular hydrogen bonds, intramolecular electrostatic interaction energy, or degree of burial of the hydrophobic surface; though the empirical rule mentioned above held, it was not the proximate cause of higher stability; and the number of van der Waals contacts NvdW, or equivalently, the intramolecular van der Waals interaction energy was an important factor governing the stability. Here we revisit the wild type and its nine mutants of staphylococcal nuclease using our statistical-mechanical theory for hydration of a protein. A molecular model is employed for water. We show that the pivotal factor is the magnitude of the water-entropy gain upon folding. The gain originates from an increase in the total volume available to the translational displacement of water molecules coexisting with the protein in the system. The magnitude is highly correlated with the denaturation temperature Tm. Moreover, the apparent correlation between NvdW and Tm as well as the empirical rule is interpretable (i.e., their physicochemical meanings can be clarified) on the basis of the water-entropy effect. PMID:27493849

  11. Light-harvesting mutants show differential gene expression upon shift to high light as a consequence of photosynthetic redox and reactive oxygen species metabolism.

    PubMed

    Tikkanen, Mikko; Gollan, Peter J; Mekala, Nageswara Rao; Isojärvi, Janne; Aro, Eva-Mari

    2014-04-19

    The amount of light energy that is harvested and directed to the photosynthetic machinery is regulated in order to control the production of reactive oxygen species (ROS) in leaf tissues. ROS have important roles as signalling factors that instigate and mediate a range of cellular responses, suggesting that the mechanisms regulating light-harvesting and photosynthetic energy transduction also affect cell signalling. In this study, we exposed wild-type (WT) Arabidopsis and mutants impaired in the regulation of photosynthetic light-harvesting (stn7, tap38 and npq4) to transient high light (HL) stress in order to study the role of these mechanisms for up- and downregulation of gene expression under HL stress. The mutants, all of which have disturbed regulation of excitation energy transfer and distribution, responded to transient HL treatment with surprising similarity to the WT in terms of general 'abiotic stress-regulated' genes associated with hydrogen peroxide and 12-oxo-phytodienoic acid signalling. However, we identified distinct expression profiles in each genotype with respect to induction of singlet oxygen and jasmonic acid-dependent responses. The results of this study suggest that the control of excitation energy transfer interacts with hormonal regulation. Furthermore, the photosynthetic pigment-protein complexes appear to operate as receptors that sense the energetic balance between the photosynthetic light reactions and downstream metabolism.

  12. Immune escape mutants of Highly Pathogenic Avian Influenza H5N1 selected using polyclonal sera: identification of key amino acids in the HA protein.

    PubMed

    Sitaras, Ioannis; Kalthoff, Donata; Beer, Martin; Peeters, Ben; de Jong, Mart C M

    2014-01-01

    Evolution of Avian Influenza (AI) viruses--especially of the Highly Pathogenic Avian Influenza (HPAI) H5N1 subtype--is a major issue for the poultry industry. HPAI H5N1 epidemics are associated with huge economic losses and are sometimes connected to human morbidity and mortality. Vaccination (either as a preventive measure or as a means to control outbreaks) is an approach that splits the scientific community, due to the risk of it being a potential driving force in HPAI evolution through the selection of mutants able to escape vaccination-induced immunity. It is therefore essential to study how mutations are selected due to immune pressure. To this effect, we performed an in vitro selection of mutants from HPAI A/turkey/Turkey/1/05 (H5N1), using immune pressure from homologous polyclonal sera. After 42 rounds of selection, we identified 5 amino acid substitutions in the Haemagglutinin (HA) protein, most of which were located in areas of antigenic importance and suspected to be prone to selection pressure. We report that most of the mutations took place early in the selection process. Finally, our antigenic cartography studies showed that the antigenic distance between the selected isolates and their parent strain increased with passage number.

  13. One High-School Class Yields 8 PhDs and 2 Nobel Laureates in Physics.

    ERIC Educational Resources Information Center

    Physics Today, 1983

    1983-01-01

    Of the 718 graduates of the class of 1950, Bronx High School of Science, at least eight members became PhD physicists (including two Nobel prize winners). A list of these PhD physicists and comments about the school made at a reunion held in New York City (June 1982) are provided. (JN)

  14. High-Resolution Mapping of Homologous Recombination Events in rad3 Hyper-Recombination Mutants in Yeast

    PubMed Central

    Dominska, Margaret; Moriel-Carretero, María; Herrera-Moyano, Emilia; Aguilera, Andrés; Petes, Thomas D.

    2016-01-01

    The Saccharomyces cerevisae RAD3 gene is the homolog of human XPD, an essential gene encoding a DNA helicase of the TFIIH complex involved in both nucleotide excision repair (NER) and transcription. Some mutant alleles of RAD3 (rad3-101 and rad3-102) have partial defects in DNA repair and a strong hyper-recombination (hyper-Rec) phenotype. Previous studies showed that the hyper-Rec phenotype associated with rad3-101 and rad3-102 can be explained as a consequence of persistent single-stranded DNA gaps that are converted to recombinogenic double-strand breaks (DSBs) by replication. The systems previously used to characterize the hyper-Rec phenotype of rad3 strains do not detect the reciprocal products of mitotic recombination. We have further characterized these events using a system in which the reciprocal products of mitotic recombination are recovered. Both rad3-101 and rad3-102 elevate the frequency of reciprocal crossovers about 100-fold. Mapping of these events shows that three-quarters of these crossovers reflect DSBs formed at the same positions in both sister chromatids (double sister-chromatid breaks, DSCBs). The remainder reflects DSBs formed in single chromatids (single chromatid breaks, SCBs). The ratio of DSCBs to SCBs is similar to that observed for spontaneous recombination events in wild-type cells. We mapped 216 unselected genomic alterations throughout the genome including crossovers, gene conversions, deletions, and duplications. We found a significant association between the location of these recombination events and regions with elevated gamma-H2AX. In addition, there was a hotspot for deletions and duplications at the IMA2 and HXT11 genes near the left end of chromosome XV. A comparison of these data with our previous analysis of spontaneous mitotic recombination events suggests that a sub-set of spontaneous events in wild-type cells may be initiated by incomplete NER reactions, and that DSCBs, which cannot be repaired by sister

  15. Miniature high-throughput chemosensing of yield, ee, and absolute configuration from crude reaction mixtures

    PubMed Central

    Bentley, Keith W.; Zhang, Peng; Wolf, Christian

    2016-01-01

    High-throughput experimentation (HTE) has emerged as a widely used technology that accelerates discovery and optimization processes with parallel small-scale reaction setups. A high-throughput screening (HTS) method capable of comprehensive analysis of crude asymmetric reaction mixtures (eliminating product derivatization or isolation) would provide transformative impact by matching the pace of HTE. We report how spontaneous in situ construction of stereodynamic metal probes from readily available, inexpensive starting materials can be applied to chiroptical chemosensing of the total amount, enantiomeric excess (ee), and absolute configuration of a wide variety of amines, diamines, amino alcohols, amino acids, carboxylic acids, α-hydroxy acids, and diols. This advance and HTS potential are highlighted with the analysis of 1 mg of crude reaction mixtures of a catalytic asymmetric reaction. This operationally simple assay uses a robust mix-and-measure protocol, is amenable to microscale platforms and automation, and provides critical time efficiency and sustainability advantages over traditional serial methods. PMID:26933684

  16. High Ligation of Inferior Mesenteric Artery in Left Colonic and Rectal Cancers: Lymph Node Yield and Survival Benefit.

    PubMed

    Charan, Ishwar; Kapoor, Akhil; Singhal, Mukesh Kumar; Jagawat, Namrata; Bhavsar, Deepak; Jain, Vikas; Kumar, Vanita; Kumar, Harvindra Singh

    2015-12-01

    During surgery for colorectal cancer, the inferior mesenteric artery (IMA) may be ligated either directly at the origin of the IMA from the aorta (high ligation) or at a point just below the origin of the left colic artery (low ligation). Sixty patients of left colonic and rectal cancer undergoing elective curative surgery in 2007 and 2008 were selected for this observational study. The resected lymph nodes were grouped into three levels: along the bowel wall (D1), along IMA below left colic (D2), and along the IMA and its root (D3). Statistical analysis was performed with SPSS version 20.0. D2 level was involved pathologically in 20 (33.3 %) and D3 in six out of 44 (13.6 %) patients. The median nodal yield with high and low ligation were 33 and 25, respectively (p = 0.048). Median overall survival for high ligation was 62 months versus 42 months for low ligation (p = 0.190). High ligation of the IMA for rectal and left colonic cancers can improve lymph node yield, thus facilitating accurate tumor staging and thus better disease prognostication, but the survival benefit is not significant. PMID:27011519

  17. High Ligation of Inferior Mesenteric Artery in Left Colonic and Rectal Cancers: Lymph Node Yield and Survival Benefit.

    PubMed

    Charan, Ishwar; Kapoor, Akhil; Singhal, Mukesh Kumar; Jagawat, Namrata; Bhavsar, Deepak; Jain, Vikas; Kumar, Vanita; Kumar, Harvindra Singh

    2015-12-01

    During surgery for colorectal cancer, the inferior mesenteric artery (IMA) may be ligated either directly at the origin of the IMA from the aorta (high ligation) or at a point just below the origin of the left colic artery (low ligation). Sixty patients of left colonic and rectal cancer undergoing elective curative surgery in 2007 and 2008 were selected for this observational study. The resected lymph nodes were grouped into three levels: along the bowel wall (D1), along IMA below left colic (D2), and along the IMA and its root (D3). Statistical analysis was performed with SPSS version 20.0. D2 level was involved pathologically in 20 (33.3 %) and D3 in six out of 44 (13.6 %) patients. The median nodal yield with high and low ligation were 33 and 25, respectively (p = 0.048). Median overall survival for high ligation was 62 months versus 42 months for low ligation (p = 0.190). High ligation of the IMA for rectal and left colonic cancers can improve lymph node yield, thus facilitating accurate tumor staging and thus better disease prognostication, but the survival benefit is not significant.

  18. The Yield of Community-Based “Retrospective” Tuberculosis Contact Investigation in a High Burden Setting in Ethiopia

    PubMed Central

    2016-01-01

    Objective To determine the yield and determinants of retrospective TB contact investigation in selected zones in Ethiopia. Materials and Methods This was a community-based cross-sectional study conducted during June-October 2014.Trained lay providers performed symptom screening for close contacts of index cases with all types of TB registered for anti-TB treatment within the last three years. We used logistic regression to determine factors associated with TB diagnosis among the contacts. Results Of 272,441 close contacts of 47, 021 index cases screened, 13,886 and 2, 091 had presumptive and active TB respectively. The yield of active TB was thus 768/100, 000, contributing 25.4% of the 7,954 TB cases reported from the study zones over the study period. The yield was highest among workplace contacts (12,650/100, 000). Active TB was twice more likely among contacts whose index cases had been registered for TB treatment within the last 12 months compared with those who had been registered 24 or more months earlier (adjusted odds ratio, AOR: 1.77 95% CI 1.42–2.21). Sex or clinical type of TB in index cases was not associated with the yield. Smear negative (SS-) index cases (AOR: 1.74 955 CI 1.13–2.68), having index cases who registered for treatment within <12 months (AOR: 2.41 95% CI 1.51–3.84) and being household contact (AOR: 0.072 95% CI 0.01–0.52) were associated with the occurrence of active TB in children. Conclusions The yield of retrospective contact investigation was about six times the case notification in the study zones, contributing a fourth of all TB cases notified over the same period. The yield was highest among workplace contacts and in those with recent past history of contact. Retrospective contact screening can serve as additional strategy to identify high risk groups not addressed through currently recommended screening approaches. PMID:27483160

  19. High-yield isolation of extracellular vesicles using aqueous two-phase system.

    PubMed

    Shin, Hyunwoo; Han, Chungmin; Labuz, Joseph M; Kim, Jiyoon; Kim, Jongmin; Cho, Siwoo; Gho, Yong Song; Takayama, Shuichi; Park, Jaesung

    2015-01-01

    Extracellular vesicles (EVs) such as exosomes and microvesicles released from cells are potential biomarkers for blood-based diagnostic applications. To exploit EVs as diagnostic biomarkers, an effective pre-analytical process is necessary. However, recent studies performed with blood-borne EVs have been hindered by the lack of effective purification strategies. In this study, an efficient EV isolation method was developed by using polyethylene glycol/dextran aqueous two phase system (ATPS). This method provides high EV recovery efficiency (~70%) in a short time (~15 min). Consequently, it can significantly increase the diagnostic applicability of EVs. PMID:26271727

  20. High Yield Research Opportunities in Geriatric Emergency Medicine: Prehospital Care, Delirium, Adverse Drug Events, and Falls

    PubMed Central

    Carpenter, Christopher R.; Shah, Manish N.; Hustey, Fredric M.; Heard, Kennon; Gerson, Lowell W.

    2011-01-01

    Emergency services constitute crucial and frequently used safety nets for older persons, an emergency visit by a senior very often indicates high vulnerability for functional decline and death, and interventions via the emergency system have significant opportunities to change the clinical course of older patients who require its services. However, the evidence base for widespread employment of emergency system-based interventions is lacking. In this article, we review the evidence and offer crucial research questions to capitalize on the opportunity to optimize health trajectories of older persons seeking emergency care in four areas: prehospital care, delirium, adverse drug events, and falls. PMID:21498881

  1. High-yield production of canine parvovirus virus-like particles in a baculovirus expression system.

    PubMed

    Jin, Hongli; Xia, Xiaohong; Liu, Bing; Fu, Yu; Chen, Xianping; Wang, Huihui; Xia, Zhenqiang

    2016-03-01

    An optimized VP2 gene from the current prevalent CPV strain (new CPV-2a) in China was expressed in a baculovirus expression system. It was found that the VP2 proteins assembled into virus-like particles (VLPs) with antigenic properties similar to those of natural CPV and with an especially high hemagglutination (HA) titer (1:2(20)). Dogs intramuscularly or orally immunized with VLPs produced antibodies against CPV with >1:80 hemagglutination inhibition (HI) units for at least 3 months. The CPV VLPs could be considered for use as a vaccine against CPV or as a platform for research on chimeric VLP vaccines against other diseases.

  2. High-yield synthesis of brookite TiO.sub.2 nanoparticles

    DOEpatents

    Huber, Dale L.; Monson, Todd C.

    2011-05-17

    A method for forming non-agglomerated brookite TiO.sub.2 nanoparticles without the use of expensive organic surfactants or high temperature processing. Embodiments of this invention use titanium isopropoxide as the titanium precursor and isopropanol as both the solvent and ligand for ligand-stabilized brookite-phase titania. Isopropanol molecules serve as the ligands interacting with the titania surfaces that stabilize the titania nanoparticles. The isopropanol ligands can be exchanged with other alcohols and other ligands during or after the nanoparticle formation reaction.

  3. Simple room-temperature preparation of high-yield large-area graphene oxide.

    PubMed

    Huang, N M; Lim, H N; Chia, C H; Yarmo, M A; Muhamad, M R

    2011-01-01

    Graphene has attracted much attention from researchers due to its interesting mechanical, electrochemical, and electronic properties. It has many potential applications such as polymer filler, sensor, energy conversion, and energy storage devices. Graphene-based nanocomposites are under an intense spotlight amongst researchers. A large amount of graphene is required for preparation of such samples. Lately, graphene-based materials have been the target for fundamental life science investigations. Despite graphene being a much sought-after raw material, the drawbacks in the preparation of graphene are that it is a challenge amongst researchers to produce this material in a scalable quantity and that there is a concern about its safety. Thus, a simple and efficient method for the preparation of graphene oxide (GO) is greatly desired to address these problems. In this work, one-pot chemical oxidation of graphite was carried out at room temperature for the preparation of large-area GO with ~100% conversion. This high-conversion preparation of large-area GO was achieved using a simplified Hummer's method from large graphite flakes (an average flake size of 500 μm). It was found that a high degree of oxidation of graphite could be realized by stirring graphite in a mixture of acids and potassium permanganate, resulting in GO with large lateral dimension and area, which could reach up to 120 μm and ~8000 μm(2), respectively. The simplified Hummer's method provides a facile approach for the preparation of large-area GO. PMID:22267928

  4. High yield production of D-xylonic acid from D-xylose using engineered Escherichia coli.

    PubMed

    Liu, Huaiwei; Valdehuesa, Kris Niño G; Nisola, Grace M; Ramos, Kristine Rose M; Chung, Wook-Jin

    2012-07-01

    An engineered Escherichia coli was constructed to produce D-xylonic acid, one of the top 30 high-value chemicals identified by US Department of Energy. The native pathway for D-xylose catabolism in E. coli W3110 was blocked by disrupting xylose isomerase (XI) and xylulose kinase (XK) genes. The native pathway for xylonic acid catabolism was also blocked by disrupting two genes both encoding xylonic acid dehydratase (yagE and yjhG). Through the introduction of a D-xylose dehydrogenase from Caulobacter crescentus, a D-xylonic acid producing E. coli was constructed. The recombinant E. coli produced up to 39.2 g L(-1) D-xylonic acid from 40 g L(-1) D-xylose in M9 minimal medium. The average productivity was as high as 1.09 g L(-1) h(-1) and no gluconic acid byproduct was produced. These results suggest that the engineered E. coli has a promising application for the industrial-scale production of D-xylonic acid.

  5. Rapid high-yield mRNA extraction for reverse-transcription PCR.

    PubMed

    Wang, Chengming; Kim, Teayoun; Gao, Dongya; Vaglenov, Alexander; Kaltenboeck, Bernhard

    2007-04-10

    Reverse-transcription PCR (RT-PCR) is the gold standard for mRNA quantification. Efficient, rapid, and high-throughput mRNA extraction is a prerequisite to ensure PCR sensitivity and precision, particularly for quantification of low-abundance mRNAs, and for large numbers of samples. Many mRNA extraction methods entail meticulous handling of individual samples, and are not well suited for large sample numbers. To achieve simple separation of mRNA binding matrix and the medium from which mRNA is to be isolated, oligo (dT)(20)-coated silica beads were used. Simple centrifugation and decanting steps can be used throughout the extraction procedure to separate supernatant fluids from the silica beads. DNase treatment reduced clumping of sedimented beads, thus facilitating bead resuspension and avoiding repeated agitation. DNase treatment also significantly reduced contaminating DNA, increased mRNA purity, and enhanced mRNA PCR readout by approximately 5-fold. The number of target transcripts per sample aliquot was higher in DNase-treated mRNA than in non-treated mRNA or in total nucleic acids. Thus, use of DNase-treated mRNA increased sensitivity of detection and quantification of low-copy transcripts. In conclusion, we describe here a simple, rapid, and cost-effective method that facilitates convenient extraction of high-quality mRNA by minimizing cumbersome mechanical disruption and pipetting steps.

  6. High yield production of D-xylonic acid from D-xylose using engineered Escherichia coli.

    PubMed

    Liu, Huaiwei; Valdehuesa, Kris Niño G; Nisola, Grace M; Ramos, Kristine Rose M; Chung, Wook-Jin

    2012-07-01

    An engineered Escherichia coli was constructed to produce D-xylonic acid, one of the top 30 high-value chemicals identified by US Department of Energy. The native pathway for D-xylose catabolism in E. coli W3110 was blocked by disrupting xylose isomerase (XI) and xylulose kinase (XK) genes. The native pathway for xylonic acid catabolism was also blocked by disrupting two genes both encoding xylonic acid dehydratase (yagE and yjhG). Through the introduction of a D-xylose dehydrogenase from Caulobacter crescentus, a D-xylonic acid producing E. coli was constructed. The recombinant E. coli produced up to 39.2 g L(-1) D-xylonic acid from 40 g L(-1) D-xylose in M9 minimal medium. The average productivity was as high as 1.09 g L(-1) h(-1) and no gluconic acid byproduct was produced. These results suggest that the engineered E. coli has a promising application for the industrial-scale production of D-xylonic acid. PMID:21917451

  7. Characteristics and Composition of a High Oil Yielding Castor Variety from Pakistan.

    PubMed

    Panhwar, Tarique; Mahesar, Sarfaraz Ahmed; Mahesar, Abdul Waheed; Kandhro, Aftab Ahmed; Talpur, Farah Naz; Laghari, Zahid Hussain; Chang, Abdul Sattar; Hussain Sherazi, Syed Tufail

    2016-01-01

    Keeping in view the versatile applications of castor oil in cosmetic, pharmaceutical and recently as renewable source, the present work is a step towards the commercialization of castor on large scale in Pakistan. The current study introduces a castor variety with high oil content. Initially seeds were physically examined for some physical parameters. Seed moisture, ash content and linear dimensions such as length, width and thickness were found to be 4.53%, 6.44%, 12.24 mm, 8.31 mm and 5.67 mm, respectively. For oil extraction, Soxhlet method was used which resulted in the high oil content 54.0%. For quality assessment of oil, physicochemical parameters were checked according to official standard AOCS methods and compared with ASTM specifications. The determined parameters were as follows; specific gravity 0.953 g/cm(-3), refractive index 1.431, viscosity 672.0 mPas.s, moisture content 0.32%, FFA 0.14%, IV 83.61 gI2/100 g, PV 2.25 meq/Kg and SV 186.0 mgKOH/g. Furthermore, fatty acid analysis of oil showed that, most abundant fatty acid was ricinoleic acid 94.59%, followed by palmitic 0.31%, linoleic 1.84%, oleic (n-9) 2.05%, oleic (n-10) 0.22%, stearic 0.45% and eicosenoic acid 0.53%. The detected fatty acids were compared with registered variety and varieties of other regions.

  8. Simple room-temperature preparation of high-yield large-area graphene oxide.

    PubMed

    Huang, N M; Lim, H N; Chia, C H; Yarmo, M A; Muhamad, M R

    2011-01-01

    Graphene has attracted much attention from researchers due to its interesting mechanical, electrochemical, and electronic properties. It has many potential applications such as polymer filler, sensor, energy conversion, and energy storage devices. Graphene-based nanocomposites are under an intense spotlight amongst researchers. A large amount of graphene is required for preparation of such samples. Lately, graphene-based materials have been the target for fundamental life science investigations. Despite graphene being a much sought-after raw material, the drawbacks in the preparation of graphene are that it is a challenge amongst researchers to produce this material in a scalable quantity and that there is a concern about its safety. Thus, a simple and efficient method for the preparation of graphene oxide (GO) is greatly desired to address these problems. In this work, one-pot chemical oxidation of graphite was carried out at room temperature for the preparation of large-area GO with ~100% conversion. This high-conversion preparation of large-area GO was achieved using a simplified Hummer's method from large graphite flakes (an average flake size of 500 μm). It was found that a high degree of oxidation of graphite could be realized by stirring graphite in a mixture of acids and potassium permanganate, resulting in GO with large lateral dimension and area, which could reach up to 120 μm and ~8000 μm(2), respectively. The simplified Hummer's method provides a facile approach for the preparation of large-area GO.

  9. Effect of Rht alleles on wheat grain yield and quality under high temperature and drought stress during booting and anthesis.

    PubMed

    Alghabari, Fahad; Ihsan, Muhammad Zahid; Hussain, Saddam; Aishia, Ghulam; Daur, Ihsanullah

    2015-10-01

    The present study examined the effects of gibberellin semi-sensitive reduced height (Rht) alleles on wheat grain yield and quality under high temperature and drought stress during booting and anthesis stages. Near-isogenic lines (NILs) of winter wheat (Rht (tall), Rht-B1b, Rht-D1b, Rht-B1c, Rht-8c, Rht-D1c, Rht-12) having background of Mercia and Maris Widgeon cultivars were compared under variable temperatures (day/night: 20/12, 27/19, 30/22, 33/25, 36/28, and 39/31 °C) and irrigation regimes. Pots were transferred to controlled thermal conditions (Saxcil growth chamber) during booting and anthesis stages and were maintained at field capacity (FC) or had water withheld. High temperature (>30 °C) and drought stress for seven consecutive days during booting and anthesis stages reduced the grain yield, while increased nitrogen (N) and sulphur (S) concentrations. A 50 % reduction in grain yield was fitted to have occurred at 37.4 °C for well-watered plants and at 31.4 °C for drought-stressed plants. The N and S concentrations were higher for severe dwarfs, whereas no significant differences were observed between tall and semi-dwarfs in Mercia. In the taller background (Maris Widgeon), N and S concentrations were significantly higher compared with that in Mercia. In Mercia, the severe dwarf Rht-D1c had higher Hagberg falling number (HFN) and sodium dodecyl sulphate (SDS) sedimentation volume. In both backgrounds, semi-dwarfs and severe dwarfs had higher HFN. Moreover, the SDS sedimentation volumes in Maris Widgeon were also higher than that in Mercia. Greater adaptability and improved grain quality traits suggested that severe dwarf Rht alleles are better able to enhance tolerance to high temperature and drought stress in wheat. PMID:26006072

  10. Effect of Rht alleles on wheat grain yield and quality under high temperature and drought stress during booting and anthesis.

    PubMed

    Alghabari, Fahad; Ihsan, Muhammad Zahid; Hussain, Saddam; Aishia, Ghulam; Daur, Ihsanullah

    2015-10-01

    The present study examined the effects of gibberellin semi-sensitive reduced height (Rht) alleles on wheat grain yield and quality under high temperature and drought stress during booting and anthesis stages. Near-isogenic lines (NILs) of winter wheat (Rht (tall), Rht-B1b, Rht-D1b, Rht-B1c, Rht-8c, Rht-D1c, Rht-12) having background of Mercia and Maris Widgeon cultivars were compared under variable temperatures (day/night: 20/12, 27/19, 30/22, 33/25, 36/28, and 39/31 °C) and irrigation regimes. Pots were transferred to controlled thermal conditions (Saxcil growth chamber) during booting and anthesis stages and were maintained at field capacity (FC) or had water withheld. High temperature (>30 °C) and drought stress for seven consecutive days during booting and anthesis stages reduced the grain yield, while increased nitrogen (N) and sulphur (S) concentrations. A 50 % reduction in grain yield was fitted to have occurred at 37.4 °C for well-watered plants and at 31.4 °C for drought-stressed plants. The N and S concentrations were higher for severe dwarfs, whereas no significant differences were observed between tall and semi-dwarfs in Mercia. In the taller background (Maris Widgeon), N and S concentrations were significantly higher compared with that in Mercia. In Mercia, the severe dwarf Rht-D1c had higher Hagberg falling number (HFN) and sodium dodecyl sulphate (SDS) sedimentation volume. In both backgrounds, semi-dwarfs and severe dwarfs had higher HFN. Moreover, the SDS sedimentation volumes in Maris Widgeon were also higher than that in Mercia. Greater adaptability and improved grain quality traits suggested that severe dwarf Rht alleles are better able to enhance tolerance to high temperature and drought stress in wheat.

  11. One-pot, high-yield synthesis of size-controlled gold particles with narrow size distribution.

    PubMed

    Guo, Shaojun; Wang, Erkang

    2007-08-01

    Here, we first report a facile one-step one-phase synthetic route to achieve size-controlled gold micro/nanoparticles with narrow size distribution by using o-diaminobenzene as a reducing agent in the presence of poly(N-vinyl-2-pyrrolidone) via a simple wet-chemical approach. All experimental data including that from scanning-electron microscopy, energy-dispersive X-ray spectroscopy, X-ray photoelectron spectroscopy, and X-ray diffraction techniques indicates that the gold micro/nanoparticles with a narrow size distribution were produced in high yield (approximately 100%).

  12. Diagnostic Yield of Chromosomal Microarray Analysis in a Cohort of Patients with Autism Spectrum Disorders from a Highly Consanguineous Population.

    PubMed

    Al-Mamari, Watfa; Al-Saegh, Abeer; Al-Kindy, Adila; Bruwer, Zandre; Al-Murshedi, Fathiya; Al-Thihli, Khalid

    2015-08-01

    Autism Spectrum Disorders are a complicated group of disorders characterized with heterogeneous genetic etiologies. The genetic investigations for this group of disorders have expanded considerably over the past decade. In our study we designed a tired approach and studied the diagnostic yield of chromosomal microarray analysis on patients referred to the Genetic and Developmental Medicine clinic in Sultan Qaboos University in Oman for autism spectrum disorders in a highly consanguineous population. Copy number variants were seen in 27% of our studied cohort of patients and it was strongly associated with dysmorphic features and congenital anomalies. PMID:25703031

  13. APPLICATIONS OF SOLVENT EXTRACTION IN THE HIGH-YIELD MULTI-PROCESS REDUCTION/SEPARATION OF Eu FROM EXCESS Sm

    SciTech Connect

    Schwantes, Jon M.; Sudowe, Ralf; Nitsche, Heino; Hoffman, Darleane C.

    2008-05-15

    A novel multi-process method for separating Eu from neighbouring lanthanides (Ln) has been developed that chemically reduces Eu(III) to Eu(II) prior to solvent extraction of Ln(III) with thenoyltrifluoroacetone in benzene. This method is capable of achieving higher purities (>99%) and separation yields than previously published multi-process methods that stabilize and separate the reduced Eu(II) as a sulphate solid and is ideal for enriching materials of high-value. Results from a variety of combinations of a chemical or electrochemical reduction process preceding a separation process using either ion exchange chromatography, reversed phase chromatography, or solvent extraction are discussed.

  14. High yield formation of lipid bilayer shells around silicon nanowires in aqueous solution.

    PubMed

    Römhildt, Lotta; Gang, Andreas; Baraban, Larysa; Opitz, Jörg; Cuniberti, Gianaurelio

    2013-09-01

    The combination of nanoscaled materials and biological self-assembly is a key step for the development of novel approaches for biotechnology and bionanoelectronic devices. Here we propose a route to merge these two subsystems and report on the formation of highly concentrated aqueous solutions of silanized silicon nanowires wrapped in a lipid bilayer shell. We developed protocols and investigated the dynamics of lipid films on both planar surfaces and silicon nanowires using fluorescence recovery after photobleaching, demonstrating fully intact and fluid bilayers without the presence of a lipid molecule reservoir. Finally, the experimental setup allowed for in situ observation of spontaneous bilayer formation around the nanowire by lipid diffusion from a vesicle to the nanowire. Such aqueous solutions of lipid coated nanowires are a versatile tool for characterization purposes and are relevant for newly emerging bioinspired electronics and nanosensorics.

  15. Some effects of water quality on the performance of high yielding cows in an arid climate.

    PubMed

    Challis, D J; Zeinstra, J S; Anderson, M J

    1987-01-01

    The water from wells in Saudi Arabia is often high in mineral content. The cows in this study had been receiving water containing between 4000 and 5000 ppm total dissolved solids. Four groups of 16 cows were calved down and milked under similar conditions in the hot summer months. Two of the groups were given normal well water and the other two groups received water after desalination in a reverse osmosis plant. One group on each type of water also had their water cooled. The groups of cows receiving treated water drank more water, consumed more concentrate and produced significantly more milk than the groups given normal well water. A similar effect on feed intake and milk production was seen when treated water was given to a 1000-cow unit with a similar number of control cows.