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Sample records for high yielding mutants

  1. 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

    2006-04-05

    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.

  2. 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.

  3. High yield of B-branch electron transfer in a quadruple reaction center mutant of the photosynthetic bacterium Rhodobacter sphaeroides.

    PubMed

    de Boer, Arjo L; Neerken, Sieglinde; de Wijn, Rik; Permentier, Hjalmar P; Gast, Peter; Vijgenboom, Erik; Hoff, Arnold J

    2002-03-05

    A new reaction center (RC) quadruple mutant, called LDHW, of Rhodobacter sphaeroides is described. This mutant was constructed to obtain a high yield of B-branch electron transfer and to study P(+)Q(B)(-) formation via the B-branch. The A-branch of the mutant RC contains two monomer bacteriochlorophylls, B(A) and beta, as a result of the H mutation L(M214)H. The latter bacteriochlorophyll replaces bacteriopheophytin H(A) of wild-type RCs. As a result of the W mutation A(M260)W, the A-branch does not contain the ubiquinone Q(A); this facilitates the study of P(+)Q(B)(-) formation. Furthermore, the D mutation G(M203)D introduces an aspartic acid residue near B(A). Together these mutations impede electron transfer through the A-branch. The B-branch contains two bacteriopheophytins, Phi(B) and H(B), and a ubiquinone, Q(B.) Phi(B) replaces the monomer bacteriochlorophyll B(B) as a result of the L mutation H(M182)L. In the LDHW mutant we find 35-45% B-branch electron transfer, the highest yield reported so far. Transient absorption spectroscopy at 10 K, where the absorption bands due to the Q(X) transitions of Phi(B) and H(B) are well resolved, shows simultaneous bleachings of both absorption bands. Although photoreduction of the bacteriopheophytins occurs with a high yield, no significant (approximately 1%) P(+)Q(B)(-) formation was found.

  4. 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

  5. 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).

  6. 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.

  7. High-yield production of aryl alcohol oxidase under limited growth conditions in small-scale systems using a mutant Aspergillus nidulans strain.

    PubMed

    Pardo-Planas, Oscar; Prade, Rolf A; Wilkins, Mark R

    2017-02-01

    Aryl alcohol oxidase (MtGloA) is an enzyme that belongs to the ligninolytic consortium and can play an important role in the bioenergy industry. This study investigated production of an MtGloA client enzyme by a mutant strain of Aspergillus nidulans unable to synthesize its own pyridoxine. Pyridoxine limitation can be used to control cell growth, diverting substrate to protein production. In agitated culture, enzyme production was similar when using media with 1 mg/L and without pyridoxine (26.64 ± 6.14 U/mg mycelia and 26.14 ± 8.39 U/mg mycelia using media with and without pyridoxine, respectively). However, the treatment lacking pyridoxine had to be supplemented with pyridoxine after 156 h of fermentation to sustain continued enzyme production. Use of extremely diluted pyridoxine levels allowed reduced fungal growth while maintaining steady enzyme production. Concentrations of 9 and 13.5 µg/L pyridoxine allowed MtGloA production with a growth rate of only 5% of that observed when using the standard 1 mg/L pyridoxine media.

  8. 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,…

  9. 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.

  10. 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

  11. Improved sugar yields from biomass sorghum feedstocks: comparing low-lignin mutants and pretreatment chemistries

    DOE PAGES

    Godin, Bruno; Nagle, Nick; Sattler, Scott; ...

    2016-11-21

    For biofuel production processes to be economically efficient, it is essential to maximize the production of monomeric carbohydrates from the structural carbohydrates of feedstocks. One strategy for maximizing carbohydrate production is to identify less recalcitrant feedstock cultivars by performing some type of experimental screening on a large and diverse set of candidate materials, or by identifying genetic modifications (random or directed mutations or transgenic plants) that provide decreased recalcitrance. Economic efficiency can also be increased using additional pretreatment processes such as deacetylation, which uses dilute NaOH to remove the acetyl groups of hemicellulose prior to dilute acid pretreatment. In thismore » work, we used a laboratory-scale screening tool that mimics relevant thermochemical pretreatment conditions to compare the total sugar yield of three near-isogenic brown midrib (bmr) mutant lines and the wild-type (WT) sorghum cultivar. We then compared results obtained from the laboratory-scale screening pretreatment assay to a large-scale pretreatment system. After pretreatment and enzymatic hydrolysis, the bmr mutants had higher total sugar yields than the WT sorghum cultivar. Increased pretreatment temperatures increased reactivity for all sorghum samples reducing the differences observed at lower reaction temperatures. Deacetylation prior to dilute acid pretreatment increased the total sugar yield for all four sorghum samples, and reduced the differences in total sugar yields among them, but solubilized a sizable fraction of the non-structural carbohydrates. The general trends of increased total sugar yield in the bmr mutant compared to the WT seen at the laboratory scale were observed at the large-scale system. However, in the larger reactor system, the measured total sugar yields were lower and the difference in total sugar yield between the WT and bmr sorghum was larger. Sorghum bmr mutants, which have a reduced lignin content

  12. Improved sugar yields from biomass sorghum feedstocks: comparing low-lignin mutants and pretreatment chemistries

    SciTech Connect

    Godin, Bruno; Nagle, Nick; Sattler, Scott; Agneessens, Richard; Delcarte, Jérôme; Wolfrum, Edward

    2016-11-21

    For biofuel production processes to be economically efficient, it is essential to maximize the production of monomeric carbohydrates from the structural carbohydrates of feedstocks. One strategy for maximizing carbohydrate production is to identify less recalcitrant feedstock cultivars by performing some type of experimental screening on a large and diverse set of candidate materials, or by identifying genetic modifications (random or directed mutations or transgenic plants) that provide decreased recalcitrance. Economic efficiency can also be increased using additional pretreatment processes such as deacetylation, which uses dilute NaOH to remove the acetyl groups of hemicellulose prior to dilute acid pretreatment. In this work, we used a laboratory-scale screening tool that mimics relevant thermochemical pretreatment conditions to compare the total sugar yield of three near-isogenic brown midrib (bmr) mutant lines and the wild-type (WT) sorghum cultivar. We then compared results obtained from the laboratory-scale screening pretreatment assay to a large-scale pretreatment system. After pretreatment and enzymatic hydrolysis, the bmr mutants had higher total sugar yields than the WT sorghum cultivar. Increased pretreatment temperatures increased reactivity for all sorghum samples reducing the differences observed at lower reaction temperatures. Deacetylation prior to dilute acid pretreatment increased the total sugar yield for all four sorghum samples, and reduced the differences in total sugar yields among them, but solubilized a sizable fraction of the non-structural carbohydrates. The general trends of increased total sugar yield in the bmr mutant compared to the WT seen at the laboratory scale were observed at the large-scale system. However, in the larger reactor system, the measured total sugar yields were lower and the difference in total sugar yield between the WT and bmr sorghum was larger. Sorghum bmr mutants, which have a reduced lignin content showed

  13. Identification of potentially high yielding irradiated cassava ‘Gajah’ genotype with different geographic coordinates

    NASA Astrophysics Data System (ADS)

    Subekti, I.; Khumaida, N.; Ardie, SW

    2017-01-01

    Cassava is one of the main and important carbohydrate producing crops in Indonesia. Thus cassava production and its tuber quality need to be improved. ‘Gajah’ genotype is a local genotypes cassava from East Kalimantan, has high potential yield (> 60 ton Ha-1). However, the harvest time of this genotype is quite long (>= 12 months). The objective of this research was to identify the high yielding cassava mutants from the gamma rays irradiated ‘Gajah’ genotype at M1V3 population and potential yield at different location. Several putative cassava mutants (12 mutants) were planted in Cikabayan Experimental Field, IPB from March 2015 to March 2016 and the yields compared with the same genotype grown at different location by seeing its coordinates to observe the potential yield. Our result showed that the fresh tuber weight per plant of some putative mutants could reach more than 8 kg (yield potential of 64 ton Ha-1). The harvested tubers also had sweet flavor, although the tubers of some putative mutants were bitter. Based on previous research study, the different geographic coordinate has resulted variability on fresh tuber yield. It seems that it needs to observe the stability of ‘Gajah’- irradiated mutants in several location in Java Island.

  14. 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

  15. High yield fabrication of fluorescent nanodiamonds.

    PubMed

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

    2009-06-10

    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.

  16. High Energy Explosive Yield Enhancer Using Microencapsulation.

    DTIC Science & Technology

    The invention consists of a class of high energy explosive yield enhancers created through the use of microencapsulation techniques. The... microcapsules consist of combinations of highly reactive oxidizers that are encapsulated in either passivated inorganic fuels or inert materials and inorganic...fuels. Depending on the application, the availability of the various oxidizers and fuels within the microcapsules can be customized to increase the

  17. High-yield pulping effluent treatment technologies

    SciTech Connect

    Su, W.X.; Hsieh, J.S. . School of Chemical Engineering)

    1993-03-01

    The objective of this report is to examine the high-yield (mechanical) pulp processes with respect to environmental issues affected by the discharge of their waste streams. Various statistics are given that support the view that high-yield pulping processes will have major growth in the US regions where pulp mills are located, and sites for projects in the development phase are indicated. Conventional and innovative effluent-treatment technologies applicable to these processes are reviewed. The different types of mechanical pulping or high-yield processes are explained, and the chemical additives are discussed. The important relationship between pulp yield and measure of BOD in the effluent is graphically presented. Effluent contaminants are identified, along with other important characteristics of the streams. Current and proposed environmental limitations specifically related to mechanical pulp production are reviewed. Conventional and innovative effluent-treatment technologies are discussed, along with their principle applications, uses, advantages, and disadvantages. Sludge management and disposal techniques become an intimate part of the treatment of waste streams. The conclusion is made that conventional technologies can successfully treat effluent streams under current waste-water discharge limitations, but these systems may not be adequate when stricter standards are imposed. At present, the most important issue in the treatment of pulp-mill waste is the management and disposal of the resultant sludge.

  18. 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

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

    PubMed

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

    2016-09-26

    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.

  20. Methods for high yield production of terpenes

    DOEpatents

    Kutchan, Toni; Higashi, Yasuhiro; Feng, Xiaohong

    2017-01-03

    Provided are enhanced high yield production systems for producing terpenes in plants via the expression of fusion proteins comprising various combinations of geranyl diphosphate synthase large and small subunits and limonene synthases. Also provided are engineered oilseed plants that accumulate monoterpene and sesquiterpene hydrocarbons in their seeds, as well as methods for producing such plants, providing a system for rapidly engineering oilseed crop production platforms for terpene-based biofuels.

  1. 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.

  2. 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

  3. Field evaluation of mint mutant and hybrid lines for resistance to Verticillium wilt and yield

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Severity of Verticillium wilt varied significantly among mint lines and cultivars in the inoculated and non-inoculated sub-plots in two field trials. Verticillium wilt was significantly less severe for mutant lines 87M0109-1, 84M0107-7, and M90-11 than for Black Mitcham in 2002 and 2003. Verticilli...

  4. High-throughput fluorescence-activated cell sorting for lipid hyperaccumulating Chlamydomonas reinhardtii mutants.

    PubMed

    Xie, Bo; Stessman, Dan; Hart, Jason H; Dong, Haili; Wang, Yingjun; Wright, David A; Nikolau, Basil J; Spalding, Martin H; Halverson, Larry J

    2014-09-01

    The genetically tractable microalga Chlamydomonas reinhardtii has many advantages as a model for renewable bioproducts and/or biofuels production. However, one limitation of C. reinhardtii is its relatively low-lipid content compared with some other algal species. To overcome this limitation, we combined ethane methyl sulfonate mutagenesis with fluorescence-activated cell sorting (FACS) of cells stained with the lipophilic stain Nile Red to isolate lipid hyperaccumulating mutants of C. reinhardtii. By manipulating the FACS gates, we sorted mutagenized cells with extremely high Nile Red fluorescence signals that were rarely detected in nonmutagenized populations. This strategy successfully isolated several putative lipid hyperaccumulating mutants exhibiting 23% to 58% (dry weight basis) higher fatty acid contents than their progenitor strains. Significantly, for most mutants, nitrogen starvation was not required to attain high-lipid content nor was there a requirement for a deficiency in starch accumulation. Microscopy of Nile Red stained cells revealed that some mutants exhibit an increase in the number of lipid bodies, which correlated with TLC analysis of triacyglycerol content. Increased lipid content could also arise through increased biomass production. Collectively, our findings highlight the ability to enhance intracellular lipid accumulation in algae using random mutagenesis in conjunction with a robust FACS and lipid yield verification regime. Our lipid hyperaccumulating mutants could serve as a genetic resource for stacking additional desirable traits to further increase lipid production and for identifying genes contributing to lipid hyperaccumulation, without lengthy lipid-induction periods.

  5. Diode laser welding of high yield steel

    NASA Astrophysics Data System (ADS)

    Lisiecki, Aleksander

    2013-01-01

    The following article describes results of investigations on influence of laser welding parameters on the weld shape, quality and mechanical properties of 2.5 mm thick butt joints of thermo-mechanically rolled, high yield strength steel for cold forming S420MC (according to EN 10149 - 3 and 060XLK according to ASTM) welded with high power diode laser HPDL ROFIN SINAR DL 020 with rectangular laser beam spot and 2.2 kW output power, and 808 nm wavelength. The investigations at the initial stage were focused on detailed analysis of influence of the basic laser welding parameters such as laser power and welding speed on the shape and quality of single bead produced during bead-on-plate welding. Then the optimal parameters were chosen for laser welding of 2.5 mm thick butt joints of the thermo-mechanically rolled, high yield strength steel sheets for cold forming S420MC. The test joints were prepared as single square groove and one-side laser welded without an additional material, at a flat position. Edges of steel sheets were melted in argon atmosphere by the laser beam focused on the top joint surface. The test welded joints were investigated by visual inspection, metallographic examinations, mechanical tests such as tensile tests and bending tests. It was found that the high power diode laser may be applied successfully for one-side welding of the S420MC steel butt joints. Additionally it was found that in the optimal range of laser welding parameters the high quality joint were produced.

  6. Temperature effect on a high stearic acid sunflower mutant.

    PubMed

    Fernández-Moya, Valle; Martínez-Force, Enrique; Garcés, Rafael

    2002-01-01

    Vegetable oil with elevated saturated fatty acid content may be useful for producing solid fat without hydrogenation or transesterification. Under the nutritional point of view stearic acid is preferred to other saturated fatty acids because of its neutral effect on serum cholesterol lipoproteins. Selection of a very high stearic acid sunflower (Helianthus annuus L.) line (CAS-14), with up to a 37.3% of stearic acid in the seed oil, and the relationship between the expression of this character and the growth temperature are presented. The mutant was selected from the M(2) progeny of 3000 mutagenized seeds (4 mM sodium azide mutagenesis treatment) by analysing the fatty acid composition of half-seed by gas liquid chromatography. In order to genetically fix the mutant character, plants were grown at high day/night temperatures during seed formation. We found that temperatures higher than 30/20 degrees C are required for good expression of the phenotype, the maximum stearic acid content being obtained at 39/24 degrees C. This behaviour is totally opposed to that observed in normal and previously isolated high-stearic acid sunflower lines that contain more stearic acid at low temperature. Thus, a new type of temperature regulation on the stearate desaturation must occur. This line is the sunflower mutant with the highest stearic acid content reported so far.

  7. Development of high-yield influenza B virus vaccine viruses.

    PubMed

    Ping, Jihui; Lopes, Tiago J S; Neumann, Gabriele; Kawaoka, Yoshihiro

    2016-12-20

    The burden of human infections with influenza A and B viruses is substantial, and the impact of influenza B virus infections can exceed that of influenza A virus infections in some seasons. Over the past few decades, viruses of two influenza B virus lineages (Victoria and Yamagata) have circulated in humans, and both lineages are now represented in influenza vaccines, as recommended by the World Health Organization. Influenza B virus vaccines for humans have been available for more than half a century, yet no systematic efforts have been undertaken to develop high-yield candidates. Therefore, we screened virus libraries possessing random mutations in the six "internal" influenza B viral RNA segments [i.e., those not encoding the major viral antigens, hemagglutinin (HA) and neuraminidase NA)] for mutants that confer efficient replication. Candidate viruses that supported high yield in cell culture were tested with the HA and NA genes of eight different viruses of the Victoria and Yamagata lineages. We identified combinations of mutations that increased the titers of candidate vaccine viruses in mammalian cells used for human influenza vaccine virus propagation and in embryonated chicken eggs, the most common propagation system for influenza viruses. These influenza B virus vaccine backbones can be used for improved vaccine virus production.

  8. Photosynthesis, light use efficiency, and yield of reduced-chlorophyll soybean mutants in field conditions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Reducing chlorophyll (chl) content may improve the conversion efficiency of absorbed radiation into biomass (ec) and therefore yield in dense monoculture crops by improving light penetration and distribution within the canopy. Modeling suggests that reducing chl content may also reduce leaf temperat...

  9. Combining high biodiversity with high yields in tropical agroforests.

    PubMed

    Clough, Yann; Barkmann, Jan; Juhrbandt, Jana; Kessler, Michael; Wanger, Thomas Cherico; Anshary, Alam; Buchori, Damayanti; Cicuzza, Daniele; Darras, Kevin; Putra, Dadang Dwi; Erasmi, Stefan; Pitopang, Ramadhanil; Schmidt, Carsten; Schulze, Christian H; Seidel, Dominik; Steffan-Dewenter, Ingolf; Stenchly, Kathrin; Vidal, Stefan; Weist, Maria; Wielgoss, Arno Christian; Tscharntke, Teja

    2011-05-17

    Local and landscape-scale agricultural intensification is a major driver of global biodiversity loss. Controversially discussed solutions include wildlife-friendly farming or combining high-intensity farming with land-sparing for nature. Here, we integrate biodiversity and crop productivity data for smallholder cacao in Indonesia to exemplify for tropical agroforests that there is little relationship between yield and biodiversity under current management, opening substantial opportunities for wildlife-friendly management. Species richness of trees, fungi, invertebrates, and vertebrates did not decrease with yield. Moderate shade, adequate labor, and input level can be combined with a complex habitat structure to provide high biodiversity as well as high yields. Although livelihood impacts are held up as a major obstacle for wildlife-friendly farming in the tropics, our results suggest that in some situations, agroforests can be designed to optimize both biodiversity and crop production benefits without adding pressure to convert natural habitat to farmland.

  10. Screening and hydrogen-producing characters of a highly efficient H₂-producing mutant of Rhodovulum sulfidophilum P5.

    PubMed

    Cai, Jinling; Wang, Guangce

    2013-08-01

    In this study, transposon mutagenesis technology was utilized to enhance the hydrogen production capability of a wild marine photosynthetic bacterium Rhodovulum sulfidophilum P5. A mutant strain TH-253 that exhibited high hydrogen yield and weaker light absorption ability was screened. Under strong light conditions, the mutant produced more hydrogen than that of the WT. Under optimum light intensity (120 μmol photons/m(2)s), the mutant achieved its highest hydrogen yield (1,436 ± 44 mL H2/L, about 3.21 ± 0.10 mol H2/mol acetate), which was 40.37% higher that of the WT. In continuous operation mode, the hydrogen yield (3.59 ± 0.11 mol H2/mol acetate) and average hydrogen production rate (16.91 ± 0.46 mL H2/Lh) of the mutant were 43.40% and 45.07% higher than those of the WT, respectively. The mutant strain TH-253 may be used as an appropriate starting strain for future photosynthesis-based large scale hydrogen production.

  11. 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.

  12. Multi-seeded sorghum mutants as a novel trait to boost grain yield

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sorghum (Sorghum bicolor L. Moench) has highly branched panicles with primary, secondary, and tertiary branches. The flower branch ends with a terminal triplet spikelet, one sessile spikelet that is directly attached to the branch and two pedicellate spikelets that are attached to the branch through...

  13. Salicylic acid deficiency in NahG transgenic lines and sid2 mutants increases seed yield in the annual plant Arabidopsis thaliana.

    PubMed

    Abreu, Maria Elizabeth; Munné-Bosch, Sergi

    2009-01-01

    Salicylic acid-deficient NahG transgenic lines and sid2 mutants were used to evaluate the role of this compound in the development of the short-lived, annual plant Arabidopsis thaliana, with a particular focus on the interplay between salicylic acid and other phytohormones. Low salicylic acid levels led to increased growth, as well as to smaller abscisic acid levels and reduced damage to PSII (as indicated by F(v)/F(m) ratios) during the reproductive stages in rosette leaves of NahG transgenic lines and sid2 mutants, compared with wild-type plants. Furthermore, salicylic acid deficiency highly influenced seed yield and composition. Seed production increased by 4.4-fold and 3.5-fold in NahG transgenic lines and sid2 mutants, respectively, compared to the wild type. Salicylic acid deficiency also improved seed composition in terms of antioxidant vitamin concentrations, seeds of salicylic acid-deficient plants showing higher levels of alpha- and gamma-tocopherol (vitamin E) and beta-carotene (pro-vitamin A) than seeds of wild-type plants. Seeds of salicylic acid-deficient plants also showed higher nitrogen concentrations than seeds of wild-type plants. It is concluded that (i) the sid2 gene, which encodes for isochorismate synthase, plays a central role in salicylic acid biosynthesis during plant development in A. thaliana, (ii) salicylic acid plays a role in the regulation of growth, senescence, and seed production, (iii) there is a cross-talk between salicylic acid and other phytohormones during plant development, and (iv) the concentrations of antioxidant vitamins in seeds may be influenced by the endogenous levels of salicylic acid in plants.

  14. Repairing the Sickle Cell mutation. II. Effect of psoralen linker length on specificity of formation and yield of third strand-directed photoproducts with the mutant target sequence.

    PubMed

    Amosova, Olga; Broitman, Steven L; Fresco, Jacques R

    2003-08-15

    Three identical deoxyoligonucleotide third strands with a 3'-terminal psoralen moiety attached by linkers that differ in length (N = 16, 6 and 4 atoms) and structure were examined for their ability to form triplex-directed psoralen photoproducts with both the mutant T residue of the Sickle Cell beta-globin gene and the comparable wild-type sequence in linear duplex targets. Specificity and yield of UVA (365 nm) and visible (419 nm) light-induced photoadducts were studied. The total photoproduct yield varies with the linker and includes both monoadducts and crosslinks at various available pyrimidine sites. The specificity of photoadduct formation at the desired mutant T residue site was greatly improved by shortening the psoralen linker. In particular, using the N-4 linker, psoralen interaction with the residues of the non-coding duplex strand was essentially eliminated, while modification of the Sickle Cell mutant T residue was maximized. At the same time, the proportion of crosslink formation at the mutant T residue upon UV irradiation was much greater for the N-4 linker. The photoproducts formed with the wild-type target were fully consistent with its single base pair difference. The third strand with the N-4 linker was also shown to bind to a supercoiled plasmid containing the Sickle Cell mutation site, giving photoproduct yields comparable with those observed in the linear mutant target.

  15. Repairing the Sickle Cell mutation. II. Effect of psoralen linker length on specificity of formation and yield of third strand-directed photoproducts with the mutant target sequence

    PubMed Central

    Amosova, Olga; Broitman, Steven L.; Fresco, Jacques R.

    2003-01-01

    Three identical deoxyoligonucleotide third strands with a 3′-terminal psoralen moiety attached by linkers that differ in length (N = 16, 6 and 4 atoms) and structure were examined for their ability to form triplex-directed psoralen photoproducts with both the mutant T residue of the Sickle Cell β-globin gene and the comparable wild-type sequence in linear duplex targets. Specificity and yield of UVA (365 nm) and visible (419 nm) light-induced photoadducts were studied. The total photoproduct yield varies with the linker and includes both monoadducts and crosslinks at various available pyrimidine sites. The specificity of photoadduct formation at the desired mutant T residue site was greatly improved by shortening the psoralen linker. In particular, using the N-4 linker, psoralen interaction with the residues of the non-coding duplex strand was essentially eliminated, while modification of the Sickle Cell mutant T residue was maximized. At the same time, the proportion of crosslink formation at the mutant T residue upon UV irradiation was much greater for the N-4 linker. The photoproducts formed with the wild-type target were fully consistent with its single base pair difference. The third strand with the N-4 linker was also shown to bind to a supercoiled plasmid containing the Sickle Cell mutation site, giving photoproduct yields comparable with those observed in the linear mutant target. PMID:12907706

  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. 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.

  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. Statistical optimization for enhanced yields of probiotic Bacillus coagulans and its phage resistant mutants followed by kinetic modelling of the process.

    PubMed

    Pandey, Kavita R; Joshi, Chetan; Vakil, Babu V

    2016-01-01

    Probiotics are microorganisms which when administered in adequate amounts confer health benefits to the host. A leading pharmaceutical company producing Bacillus coagulans as a probiotic was facing the problem of recurring phage attacks. Two mutants viz. B. co PIII and B. co MIII that were isolated as phage resistant mutants after UV irradiation and MMS treatment of phage sensitive B. coagulans parental culture were characterized at functional and molecular level and were noted to have undergone interesting genetic changes. The non-specific genetic alterations induced by mutagenesis can also lead to alterations in cell performance. Hence, in the current study the parental strain and the two mutants were selected for shake flask optimization. Plackett-Burman design was used to select the significant culture variables affecting biomass production. Evolutionary operation method was applied for further optimization. The study showed wide variations in the nutritional requirements of phage resistant mutants, post exposure to mutagens. An increment of 150, 134 and 152 % was observed in the biomass productions of B. coagulans (parental type) and mutants B.co PIII and B.co MIII respectively, compared to the yield from one-factor-at-a-time technique. Using Logistic and modified Leudeking-Piret equations, biomass accumulation and substrate utilization efficiency of the bioprocess were determined. The experimental data was in agreement with the results predicted by statistical analysis and modelling. The developed model may be useful for controlling the growth and substrate consumption kinetics in large scale fermentation using B. coagulans.

  20. 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.

  1. High-throughput screening of a Corynebacterium glutamicum mutant library on genomic and metabolic level.

    PubMed

    Reimer, Lorenz C; Spura, Jana; Schmidt-Hohagen, Kerstin; Schomburg, Dietmar

    2014-01-01

    Due to impressive achievements in genomic research, the number of genome sequences has risen quickly, followed by an increasing number of genes with unknown or hypothetical function. This strongly calls for development of high-throughput methods in the fields of transcriptomics, proteomics and metabolomics. Of these platforms, metabolic profiling has the strongest correlation with the phenotype. We previously published a high-throughput metabolic profiling method for C. glutamicum as well as the automatic GC/MS processing software MetaboliteDetector. Here, we added a high-throughput transposon insertion determination for our C. glutamicum mutant library. The combination of these methods allows the parallel analysis of genotype/phenotype correlations for a large number of mutants. In a pilot project we analyzed the insertion points of 722 transposon mutants and found that 36% of the affected genes have unknown functions. This underlines the need for further information gathered by high-throughput techniques. We therefore measured the metabolic profiles of 258 randomly chosen mutants. The MetaboliteDetector software processed this large amount of GC/MS data within a few hours with a low relative error of 11.5% for technical replicates. Pairwise correlation analysis of metabolites over all genotypes showed dependencies of known and unknown metabolites. For a first insight into this large data set, a screening for interesting mutants was done by a pattern search, focusing on mutants with changes in specific pathways. We show that our transposon mutant library is not biased with respect to insertion points. A comparison of the results for specific mutants with previously published metabolic results on a deletion mutant of the same gene confirmed the concept of high-throughput metabolic profiling. Altogether the described method could be applied to whole mutant libraries and thereby help to gain comprehensive information about genes with unknown, hypothetical and known

  2. Process produces chlorinated aromatic isocyanate in high yield

    NASA Technical Reports Server (NTRS)

    Trischler, F.

    1966-01-01

    Tetrachloreterephthaloyl chloride reacts with sodium azide in an atmosphere of nitrogen to form a high yield of tetrachloro-p-phenylene diisocyanate. The chlorinated diisocyanate should have application as an intermediate in the preparation of polyurethane foams. The high halogen content would impart added flame resistance to these foams.

  3. 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...

  4. [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.

  5. 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.

  6. 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.

  7. Strontium and barium iodide high light yield scintillators

    NASA Astrophysics Data System (ADS)

    Cherepy, Nerine J.; Hull, Giulia; Drobshoff, Alexander D.; Payne, Stephen A.; van Loef, Edgar; Wilson, Cody M.; Shah, Kanai S.; Roy, Utpal N.; Burger, Arnold; Boatner, Lynn A.; Choong, Woon-Seng; Moses, William W.

    2008-02-01

    Europium-doped strontium and barium iodide are found to be readily growable by the Bridgman method and to produce high scintillation light yields. SrI2(Eu ) emits into the Eu2+ band, centered at 435nm, with a decay time of 1.2μs and a light yield of ˜90000photons/MeV. It offers energy resolution better than 4% full width at half maximum at 662keV, and exhibits excellent light yield proportionality. BaI2(Eu ) produces >30000photons/MeV into the Eu2+ band at 420nm (<1μs decay). An additional broad impurity-mediated recombination band is present at 550nm (>3μs decay), unless high-purity feedstock is used.

  8. Simulation of DSB yield for high LET radiation.

    PubMed

    Friedrich, T; Durante, M; Scholz, M

    2015-09-01

    A simulation approach for the calculation of the LET-dependent yield of double-strand breaks (DSB) is presented. The model considers DSB formed as two close-lying single-strand breaks (SSB), whose formation is mediated by both intra-track processes (single electrons) or at local doses larger than about 1000 Gy in particle tracks also by electron inter-track processes (two independent electron tracks). A Monte Carlo algorithm and an analytical formula for the DSB yield are presented. The approach predicts that the DSB yield is enhanced after charged particle irradiation of high LET compared with X-ray or gamma radiation. It is used as an inherent part of the local effect model, which is applied to estimate the relative biological effectiveness of high LET radiation.

  9. High-yield positron systems for linear colliders

    SciTech Connect

    Clendenin, J.E.

    1989-04-01

    Linear colliders, such as the SLC, are among those accelerators for which a high-yield positron source operating at the repetition rate of the accelerator is desired. The SLC, having electron energies up to 50 GeV, presents the possibility of generating positron bunches with useful charge even exceeding that of the initial electron bunch. The exact positron yield to be obtained depends on the particular capture, transport and damping system employed. Using 31 GeV electrons impinging on a W-type converter phase-space at the target to the acceptance of the capture rf section, the SLC source is capable of producing, for every electron, up to two positrons within the acceptance of the positron damping ring. The design of this source and the performance of the positron system as built are described. Also, future prospects and limitations for high-yield positron systems are discussed. 11 refs., 5 figs., 3 tabs.

  10. 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.

  11. Feasibility of High Yield / High Gain NIF Capsules

    SciTech Connect

    Suter, L.; Rothenberg, J.; Munro, D.; Van Wonterghem, B.; Haan, S.; Lindl, J.

    1999-12-06

    Our original ignition ''point designs'' (circa 1992) for the National Ignition Facility (NIF) were made energetically conservative to provide margin for uncertainties in laser absorption, x-ray conversion efficiency and hohlraum-capsule coupling. Since that time, extensive experiments on Nova and Omega and their related analysis indicate that NIF coupling efficiency may be almost ''as good as we could hope for''. Given close agreement between experiment and theory/modeling, we can credibly explore target enhancements which couple more of NIF's energy to an ignition capsule. We find that 3-4X increases in absorbed capsule energy appear possible, providing a potentially more robust target and {approx}10X increase in capsule yield.

  12. 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.

  13. 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.)

  14. A new technique for measuring sputtering yields at high energies

    NASA Technical Reports Server (NTRS)

    Qiu, Y.; Griffith, J. E.; Tombrello, T. A.

    1984-01-01

    The use of thin, self-supporting carbon catcher foils allows one to measure sputtering yields in a broad range of materials with high sensitivity. Analyzing the foils with Rutherford forward scattering, sputtered Al, Si and P surface densities down to 5 x 10 to the 13th per sq cm with uncertainties of about 20 percent have been measured.

  15. 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.

  16. High-yield pulping effluent treatment technologies. Final report

    SciTech Connect

    Su, W.X.; Hsieh, J.S.

    1993-03-01

    The objective of this report is to examine the high-yield (mechanical) pulp processes with respect to environmental issues affected by the discharge of their waste streams. Various statistics are given that support the view that high-yield pulping processes will have major growth in the US regions where pulp mills are located, and sites for projects in the development phase are indicated. Conventional and innovative effluent-treatment technologies applicable to these processes are reviewed. The different types of mechanical pulping or high-yield processes are explained, and the chemical additives are discussed. The important relationship between pulp yield and measure of BOD in the effluent is graphically presented. Effluent contaminants are identified, along with other important characteristics of the streams. Current and proposed environmental limitations specifically related to mechanical pulp production are reviewed. Conventional and innovative effluent-treatment technologies are discussed, along with their principle applications, uses, advantages, and disadvantages. Sludge management and disposal techniques become an intimate part of the treatment of waste streams. The conclusion is made that conventional technologies can successfully treat effluent streams under current waste-water discharge limitations, but these systems may not be adequate when stricter standards are imposed. At present, the most important issue in the treatment of pulp-mill waste is the management and disposal of the resultant sludge.

  17. 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.

  18. 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...

  19. 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

  20. High-yield criteria for panoramic radiography. Final report

    SciTech Connect

    White, S.C.; Forsythe, A.B.

    1982-06-01

    Panoramic radiographs should be obtained when the examination offers the prospect of providing information that will assist in patient care. The purpose of this study was to determine whether high-yield criteria could be developed for the use of panoramic radiographs in the treatment planning of patients seeking dental care. Clinicians were asked what signs or symptoms caused them to order a panoramic radiograph upon patient admission into the UCLA Dental Clinic. At the time the patient was radiographed, a variety of demographic and clinical measures were recorded. The most important high-yield criterion for the panoramic examination is whether the radiograph is ordered for 'general screening examination' (a negative predictor) and whether the radiograph was ordered for any specific examination (a positive predictor). The use of these (or any other) decision rules required clinical judgment of the costs (social and economic) of a missed positive finding relative to that of an unproductive examination.

  1. CtBP1/BARS Gly172 {yields} Glu mutant structure: Impairing NAD(H)-binding and dimerization

    SciTech Connect

    Nardini, Marco; Valente, Carmen; Ricagno, Stefano; Luini, Alberto; Corda, Daniela; Bolognesi, Martino

    2009-03-27

    C-terminal binding proteins (CtBPs) are multi-functional proteins involved in nuclear transcriptional co-repression, Golgi membrane fission, and synaptic ribbon formation. Binding of NAD(H) to CtBPs promotes dimerization. CtBP dimers act as a scaffold for multimeric protein complex formation, thus bridging transcriptional repressors and their targets in the nucleus. Based on size-exclusion chromatography experiments and on the crystal structure of the NAD(H)-free G172E CtBP mutant, we show here that absence of NAD(H) induces flexibility/backbone conformational changes at the dimerization interface and at the CtBP interdomain region. The results presented shed first light on the correlation between NAD(H)-binding and functional CtBP dimerization.

  2. Ascorbate-Deficient Mutants of Arabidopsis Grow in High Light Despite Chronic Photooxidative Stress1

    PubMed Central

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

    2004-01-01

    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. PMID:14963245

  3. 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.

  4. High-yield bang time detector for the OMEGA laser

    NASA Astrophysics Data System (ADS)

    Glebov, V. Yu.; Stoeckl, C.; Sangster, T. C.; Mileham, C.; Roberts, S.; Lerche, R. A.

    2006-10-01

    A simple, low-cost, high-yield neutron bang time (HYNBT) detector has been developed and implemented on the 60-beam, 30kJ OMEGA Laser Facility at the University of Rochester's Laboratory for Laser Energetics. The HYNBT consists of three chemical-vapor deposition diamond detectors of different sizes and sensitivities placed in a lead-shielded housing. The HYNBT is located in a reentrant tube 50cm from the center of the target chamber. The HYNBT has been temporally cross calibrated against the streak-camera-based neutron temporal diagnostic (NTD) for both D2 and DT implosions. The HYNBT has an internal time resolution better than 20ps and is able to measure bang time for yields above 1010 for DT and 5×1010 for D2 implosions. The implementation of the HYNBT on the National Ignition Facility will be discussed.

  5. 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.

  6. 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

  7. Highly-substrate active isoenzyme acetylcholinesterase-II, in rosy eye mutant of Aedes aegypti mosquito.

    PubMed

    Mourya, D T; Gokhale, M D; Barde, P V; Deobagkar, D N

    2001-08-01

    Insecticide bioassays were carried out on larvae and adults of rosy eye mutant and wildtype strains of A. aegypti. Both the strains were equally susceptible to DDT, malathion and deltamethrin. Biochemical assays showed an increase in acetylcholinesterase enzyme (AChE) activity in all the stages of mutant strain with both the substrates i.e. acetylthiocholine iodide and S-butyrylthiocholine iodide. However, there was no difference in the percent inhibition of enzyme activity with propoxur in these two strains. Polyacrylamide gel electrophoresis performed in native conditions on the homogenates of adults of rosy eye mosquitoes showed that AChE-II allele was highly active with the substrate acetylthiocholine iodide as compared to wildtype strain. Frequency of the highly active AChE-II allele in the mutant strain was about 68%, whereas it was about 5% in the wildtype strain.

  8. [High throughput screening atrazine chlorohydrolase mutants with enhanced activity through Haematococcus pluvialis expression system].

    PubMed

    Wang, Huizhuan; Chen, Xiwen; Hao, Xiaohua; Chen, Defu

    2011-04-01

    Developing a high-throughput screening method is of great importance for directed evolution of atrazine chlorohydrolase. A mutagenesis library of atzA from Pseudomonas sp. ADP and Arthrobacter sp. AD1 was constructed using error-prone PCR and DNA shuffling. Candidate mutants were screened through Haematococcus pluvialis expression system, using atrazine as selection pressure. Sequence analysis showed that mutations in the obtained 12 mutants with enhanced activity were all point-substitutions and scattered throughout the gene. Enzymatic activity analysis showed that the mutants all had higher activities than that of the wild type. The activities were 1.8-3.6 fold of the wild-type enzyme when cultured in BBM medium with 1 mg/L atrazine, whereas 1.8-2.6 fold with 2 mg/L atrazine. These results indicated that Haematococcus pluvialis expression system is an ideal high throughput screening system for directed evolution of atrazine chlorohydrolase.

  9. 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.

  10. 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.

  11. Regulation of hordein synthesis in barley high lysine mutant Notch-2.

    PubMed

    Tyagi, A; Santha, I M; Mehta, S L

    1992-02-01

    Genomic DNA isolated from barley cv. NP 113 and its high lysine mutant Notch-2, and restricted with different restriction enzymes was hybridized with B1 and C-hordein DNA probes. Similar Southern hybridization patterns were observed between NP 113 and Notch-2. Dot blot hybridization analysis of RNA isolated at different developmental stages and from different tissues of seed showed temporal as well as tissue specific expression. The results obtained indicate that regulation at the level of transcription/post transcription may be responsible for lower accumulation of hordein in mutant Notch-2.

  12. 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

  13. Copper ligation to soluble oligomers of the English mutant of the amyloid-β peptide yields a linear Cu(I) site that is resistant to O2 oxidation.

    PubMed

    Peck, Kristy L; Clewett, Heather S; Schmitt, Jennifer C; Shearer, Jason

    2013-05-25

    Copper coordination to soluble oligomers of the English (AβH(6)R) mutant of the amyloid-β peptide is probed. Cu(II) coordination yields a square planar (N/O)4 coordination environment, while reduction yields an O2 inert linear bis-His Cu(I) centre.

  14. 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.

  15. 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.

  16. Listeria monocytogenes mutants with altered growth phenotypes at refrigeration temperature and high salt concentrations.

    PubMed

    Burall, Laurel S; Laksanalamai, Pongpan; Datta, Atin R

    2012-02-01

    Listeria monocytogenes can survive and grow in refrigerated temperatures and high-salt environments. In an effort to better understand the associated mechanisms, a library of ∼ 5,200 transposon mutants of LS411, a food isolate from the Jalisco cheese outbreak, were screened for their ability to grow in brain heart infusion (BHI) broth at 5°C or in the presence of 7% NaCl and two mutants with altered growth profiles were identified. The LS522 mutant has a transposon insertion between secA2 and iap and showed a significant reduction in growth in BHI broth at 5°C and in the presence of 7% NaCl. Reverse transcriptase quantitative PCR (RT-qPCR) revealed a substantial reduction in the expression of iap. Additionally, a hypothetical gene (met), containing a putative S-adenosylmethionine-dependent methyltransferase domain, downstream of iap had downregulated expression. In-frame deletion mutants of iap and met were created in LS411. The LS560 (LS411 Δiap) mutant showed reduced growth at 5°C and in the presence of 7% salt, confirming its role in cold and salt growth attenuation. Surprisingly, the LS655 (LS411 Δmet) mutant showed slightly increased growth during refrigeration, though no alteration was seen in salt growth relative to the wild-type strain. The LS527 mutant, containing an insertion 36 bp upstream of the gbu operon, showed reduced expression of the gbu transcript by RT-qPCR and also showed growth reduction at 5°C and in the presence of 7% salt. This attenuation was severely exacerbated when the mutant was grown under the combined stresses. Analysis of the gbu operon deletion mutant showed decreased growth in 7% salt and refrigeration, supporting the previously characterized role for this gene in cold and salt adaptation. These studies indicate the potential for an intricate relationship between environmental stress regulation and virulence in L. monocytogenes.

  17. High yield production of extracellular recombinant levansucrase by Bacillus megaterium.

    PubMed

    Korneli, Claudia; Biedendieck, Rebekka; David, Florian; Jahn, Dieter; Wittmann, Christoph

    2013-04-01

    In this study, a high yield production bioprocess with recombinant Bacillus megaterium for the production of the extracellular enzyme levansucrase (SacB) was developed. For basic optimization of culture parameters and nutrients, a recombinant B. megaterium reporter strain that produced green fluorescent protein under control of a vector-based xylose-inducible promoter was used. It enabled efficient microtiter plate-based screening via fluorescence analysis. A pH value of pH 6, 20 % of dissolved oxygen, 37 °C, and elevated levels of biotin (100 μg L(-1)) were found optimal with regard to high protein yield and reduced overflow metabolism. Among the different compounds tested, fructose and glycerol were identified as the preferred source of carbon. Subsequently, the settings were transferred to a B. megaterium strain recombinantly producing levansucrase SacB based on the plasmid-located xylose-inducible expression system. In shake flask culture under the optimized conditions, the novel strain already secreted the target enzyme in high amounts (14 U mL(-1) on fructose and 17.2 U mL(-1) on glycerol). This was further increased in high cell density fed-batch processes up to 55 U mL(-1), reflecting a levansucrase concentration of 0.52 g L(-1). This is 100-fold more than previous efforts for this enzyme in B. megaterium and more than 10-fold higher than reported values of other extracellular protein produced in this microorganism so far. The recombinant strain could also handle raw glycerol from biodiesel industry which provided the same amount and quality of the recombinant protein and suggests future implementation into existing biorefinery concepts.

  18. High yield neutron generators using the DD reaction

    SciTech Connect

    Vainionpaa, J. H.; Harris, J. L.; Piestrup, M. A.; Gary, C. K.; Williams, D. L.; Apodaca, M. D.; Cremer, J. T.; Ji, Qing; Ludewigt, B. A.; Jones, G.

    2013-04-19

    A product line of high yield neutron generators has been developed at Adelphi technology inc. The generators use the D-D fusion reaction and are driven by an ion beam supplied by a microwave ion source. Yields of up to 5 Multiplication-Sign 10{sup 9} n/s have been achieved, which are comparable to those obtained using the more efficient D-T reaction. The microwave-driven plasma uses the electron cyclotron resonance (ECR) to produce a high plasma density for high current and high atomic ion species. These generators have an actively pumped vacuum system that allows operation at reduced pressure in the target chamber, increasing the overall system reliability. Since no radioactive tritium is used, the generators can be easily serviced, and components can be easily replaced, providing essentially an unlimited lifetime. Fast neutron source size can be adjusted by selecting the aperture and target geometries according to customer specifications. Pulsed and continuous operation has been demonstrated. Minimum pulse lengths of 50 {mu}s have been achieved. Since the generators are easily serviceable, they offer a long lifetime neutron generator for laboratories and commercial systems requiring continuous operation. Several of the generators have been enclosed in radiation shielding/moderator structures designed for customer specifications. These generators have been proven to be useful for prompt gamma neutron activation analysis (PGNAA), neutron activation analysis (NAA) and fast neutron radiography. Thus these generators make excellent fast, epithermal and thermal neutron sources for laboratories and industrial applications that require neutrons with safe operation, small footprint, low cost and small regulatory burden.

  19. High yield neutron generators using the DD reaction

    NASA Astrophysics Data System (ADS)

    Vainionpaa, J. H.; Harris, J. L.; Piestrup, M. A.; Gary, C. K.; Williams, D. L.; Apodaca, M. D.; Cremer, J. T.; Ji, Qing; Ludewigt, B. A.; Jones, G.

    2013-04-01

    A product line of high yield neutron generators has been developed at Adelphi technology inc. The generators use the D-D fusion reaction and are driven by an ion beam supplied by a microwave ion source. Yields of up to 5 × 109 n/s have been achieved, which are comparable to those obtained using the more efficient D-T reaction. The microwave-driven plasma uses the electron cyclotron resonance (ECR) to produce a high plasma density for high current and high atomic ion species. These generators have an actively pumped vacuum system that allows operation at reduced pressure in the target chamber, increasing the overall system reliability. Since no radioactive tritium is used, the generators can be easily serviced, and components can be easily replaced, providing essentially an unlimited lifetime. Fast neutron source size can be adjusted by selecting the aperture and target geometries according to customer specifications. Pulsed and continuous operation has been demonstrated. Minimum pulse lengths of 50 μs have been achieved. Since the generators are easily serviceable, they offer a long lifetime neutron generator for laboratories and commercial systems requiring continuous operation. Several of the generators have been enclosed in radiation shielding/moderator structures designed for customer specifications. These generators have been proven to be useful for prompt gamma neutron activation analysis (PGNAA), neutron activation analysis (NAA) and fast neutron radiography. Thus these generators make excellent fast, epithermal and thermal neutron sources for laboratories and industrial applications that require neutrons with safe operation, small footprint, low cost and small regulatory burden.

  20. High temperature specifically affects the photoprotective responses of chlorophyll b-deficient wheat mutant lines.

    PubMed

    Brestic, Marian; Zivcak, Marek; Kunderlikova, Kristyna; Allakhverdiev, Suleyman I

    2016-12-01

    The effects of high temperature on CO2 assimilation rate, processes associated with photosynthetic electron and proton transport, as well as photoprotective responses, were studied in chlorophyll b-deficient mutant lines (ANK-32A and ANK-32B) and wild type (WT) of wheat (Triticum aestivum L.). Despite the low chlorophyll content and chlorophyll a-to-b ratio, the non-stressed mutant plants had the similar level of CO2 assimilation and photosynthetic responses as WT. However, in ANK mutant plants exposed to prolonged high temperature episode (42 °C for ~10 h), we observed lower CO2 assimilation compared to WT, especially when a high CO2 supply was provided. In all heat-exposed plants, we found approximately the same level of PSII photoinhibition, but the decrease in content of photooxidizable PSI was higher in ANK mutant plants compared to WT. The PSI damage can be well explained by the level of overreduction of PSI acceptor side observed in plants exposed to high temperature, which was, in turn, the result of the insufficient transthylakoid proton gradient associated with low non-photochemical quenching and lack of ability to downregulate the linear electron transport to keep the reduction state of PSI acceptor side low enough. Compared to WT, the ANK mutant lines had lower capacity to drive the cyclic electron transport around PSI in moderate and high light; it confirms the protective role of cyclic electron transport for the protection of PSI against photoinhibition. Our results, however, also suggest that the inactivation of PSI in heat stress conditions can be the protective mechanism against photooxidative damage of chloroplast and cell structures.

  1. Enhanced Priming of Adaptive Immunity by Mycobacterium smegmatis Mutants with High-Level Protein Secretion

    PubMed Central

    Taylor, Natalie; Bahunde, Faith; Thompson, Afton; Yu, Jae-Sung; Jacobs, William R.; Letvin, Norm L.; Haynes, Barton F.

    2012-01-01

    Mycobacteria have features that make them attractive as potential vaccine vectors. The nonpathogenic and rapidly growing Mycobacterium smegmatis can express both Mycobacterium tuberculosis antigens and heterologous antigens from other pathogens, and it has been used as a viable vector for the development of live vaccines. In order to further improve antigen-specific immunogenicity of M. smegmatis, we screened a random transposon mutant library for mutants displaying enhanced efficiency of protein secretion (“high secretors”) and isolated 61 mutants showing enhanced endogenic and transgenic protein secretion. Sequence analysis identified a total of 54 genes involved in optimal secretion of insert proteins, as well as multiple independent transposon insertions localized within the same genomic loci and operons. The majority of transposon insertions occurred in genes that have no known protein secretion function. These transposon mutants were shown to prime antigen-specific CD8+ T cell responses better than the parental strain. Specifically, upon introducing the simian immunodeficiency virus (SIV) gag gene into these transposon mutant strains, we observed that they primed SIV Gag-specific CD8+ T cell responses significantly better than the control prime immunization in a heterologous prime/boost regimen. Our results reveal a dependence on bacterial secretion of mycobacterial and foreign antigens for the induction of antigen-specific CD8+ T cells in vivo. The data also suggest that these M. smegmatis transposon mutants could be used as novel live attenuated vaccine strains to express foreign antigens, such as those of human immunodeficiency virus type 1 (HIV-1), and induce strong antigen-specific T cell responses. PMID:22787192

  2. High-yielding syntheses of hydrophilic, conjugatable chlorins and bacteriochlorins†

    PubMed Central

    McCarthy, Jason R.; Bhaumik, Jayeeta; Merbouh, Nabyl; Weissleder, Ralph

    2009-01-01

    SUMMARY Next-generation photodynamic therapy agents based upon the conjugation of multiple photosensitizers to a targeting backbone will allow for more efficacious light-based therapies. To this end, we have developed glucose-modified chlorins and bacteriochlorins featuring a reactive carboxylic acid linker for conjugation to targeting moities. The photosensitizers were synthesized in relatively high yields from meso-tetra(p-aminophenyl)porphyrin, and resulted in neutral, hydrophilic chromophores with superb absorption profiles in the far-red and near-infrared portions of the electromagnetic spectrum. In addition, conjugation of these photosensitizers to a model nanoscaffold (crosslinked dextran-coated nanoparticles) demonstrated that the inclusion of hydrophilic sugar moieties increased the number of dyes that can be loaded while maintaining suspension stability. The described compounds are expected to be particularly useful in the synthesis of a number of targeted nanotherapeutic systems. PMID:19675897

  3. 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.

  4. High-throughput transformation method for Yarrowia lipolytica mutant library screening.

    PubMed

    Leplat, Christophe; Nicaud, Jean-Marc; Rossignol, Tristan

    2015-09-01

    As a microorganism of major biotechnological importance, the oleaginous yeast Yarrowia lipolytica is subjected to intensive genetic engineering and functional genomic analysis. Future advancements in this area, however, require a system that will generate a large collection of mutants for high-throughput screening. Here, we report a rapid and efficient method for high-throughput transformation of Y. lipolytica in 96-well plates. We developed plasmids and strains for the large-scale screening of overexpression mutant strains, using Gateway® vectors that were adapted for specific locus integration in Y. lipolytica. As an example, a collection of mutants that overexpressed the alkaline extracellular protease (AEP) was obtained in a single transformation experiment. The platform strain that we developed to receive the overexpression cassette was designed to constitutively express a fluorescent protein as a convenient growth reporter for screening in non-translucid media. An example of growth comparison in skim milk-based medium between AEP overexpression and deletion mutants is provided.

  5. 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.

  6. 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.

  7. A natural variant of NAL1, selected in high-yield rice breeding programs, pleiotropically increases photosynthesis rate.

    PubMed

    Takai, Toshiyuki; Adachi, Shunsuke; Taguchi-Shiobara, Fumio; Sanoh-Arai, Yumiko; Iwasawa, Norio; Yoshinaga, Satoshi; Hirose, Sakiko; Taniguchi, Yojiro; Yamanouchi, Utako; Wu, Jianzhong; Matsumoto, Takashi; Sugimoto, Kazuhiko; Kondo, Katsuhiko; Ikka, Takashi; Ando, Tsuyu; Kono, Izumi; Ito, Sachie; Shomura, Ayahiko; Ookawa, Taiichiro; Hirasawa, Tadashi; Yano, Masahiro; Kondo, Motohiko; Yamamoto, Toshio

    2013-01-01

    Improvement of leaf photosynthesis is an important strategy for greater crop productivity. Here we show that the quantitative trait locus GPS (GREEN FOR PHOTOSYNTHESIS) in rice (Oryza sativa L.) controls photosynthesis rate by regulating carboxylation efficiency. Map-based cloning revealed that GPS is identical to NAL1 (NARROW LEAF1), a gene previously reported to control lateral leaf growth. The high-photosynthesis allele of GPS was found to be a partial loss-of-function allele of NAL1. This allele increased mesophyll cell number between vascular bundles, which led to thickened leaves, and it pleiotropically enhanced photosynthesis rate without the detrimental side effects observed in previously identified nal1 mutants, such as dwarf plant stature. Furthermore, pedigree analysis suggested that rice breeders have repeatedly selected the high-photosynthesis allele in high-yield breeding programs. The identification and utilization of NAL1 (GPS) can enhance future high-yield breeding and provides a new strategy for increasing rice productivity.

  8. Intermittent high-dose treatment with erlotinib enhances therapeutic efficacy in EGFR-mutant lung cancer.

    PubMed

    Schöttle, Jakob; Chatterjee, Sampurna; Volz, Caroline; Siobal, Maike; Florin, Alexandra; Rokitta, Dennis; Hinze, Yvonne; Dietlein, Felix; Plenker, Dennis; König, Katharina; Albus, Kerstin; Heuckmann, Johannes M; Rauh, Daniel; Franz, Thomas; Neumaier, Bernd; Fuhr, Uwe; Heukamp, Lukas C; Ullrich, Roland T

    2015-11-17

    Treatment with EGFR kinase inhibitors improves progression-free survival of patients with EGFR-mutant lung cancer. However, all patients with initial response will eventually acquire resistance and die from tumor recurrence. We found that intermittent high-dose treatment with erlotinib induced apoptosis more potently and improved tumor shrinkage significantly than the established low doses. In mice carrying EGFR-mutant xenografts intermittent high-dose treatment (200 mg/kg every other day) was tolerable and prolonged progression-free survival and reduced the frequency of acquired resistance. Intermittent EGFR-targeted high-dose schedules induce more profound as well as sustained target inhibition and may afford enhanced therapeutic efficacy.

  9. Genetic requirements for high constitutive SOS expression in recA730 mutants of Escherichia coli.

    PubMed

    Vlašić, Ignacija; Šimatović, Ana; Brčić-Kostić, Krunoslav

    2011-09-01

    The RecA protein in its functional state is in complex with single-stranded DNA, i.e., in the form of a RecA filament. In SOS induction, the RecA filament functions as a coprotease, enabling the autodigestion of the LexA repressor. The RecA filament can be formed by different mechanisms, but all of them require three enzymatic activities essential for the processing of DNA double-stranded ends. These are helicase, 5'-3' exonuclease, and RecA loading onto single-stranded DNA (ssDNA). In some mutants, the SOS response can be expressed constitutively during the process of normal DNA metabolism. The RecA730 mutant protein is able to form the RecA filament without the help of RecBCD and RecFOR mediators since it better competes with the single-strand binding (SSB) protein for ssDNA. As a consequence, the recA730 mutants show high constitutive SOS expression. In the study described in this paper, we studied the genetic requirements for constitutive SOS expression in recA730 mutants. Using a β-galactosidase assay, we showed that the constitutive SOS response in recA730 mutants exhibits different requirements in different backgrounds. In a wild-type background, the constitutive SOS response is partially dependent on RecBCD function. In a recB1080 background (the recB1080 mutation retains only helicase), constitutive SOS expression is partially dependent on RecBCD helicase function and is strongly dependent on RecJ nuclease. Finally, in a recB-null background, the constitutive SOS expression of the recA730 mutant is dependent on the RecJ nuclease. Our results emphasize the importance of the 5'-3' exonuclease for high constitutive SOS expression in recA730 mutants and show that RecBCD function can further enhance the excellent intrinsic abilities of the RecA730 protein in vivo.

  10. High-Throughput Parallel Sequencing to Measure Fitness of Leptospira interrogans Transposon Insertion Mutants during Acute Infection

    PubMed Central

    Matsunaga, James; Haake, David A.

    2016-01-01

    Pathogenic species of Leptospira are the causative agents of leptospirosis, a zoonotic disease that causes mortality and morbidity worldwide. The understanding of the virulence mechanisms of Leptospira spp is still at an early stage due to the limited number of genetic tools available for this microorganism. The development of random transposon mutagenesis in pathogenic strains a decade ago has contributed to the identification of several virulence factors. In this study, we used the transposon sequencing (Tn-Seq) technique, which combines transposon mutagenesis with massive parallel sequencing, to study the in vivo fitness of a pool of Leptospira interrogans mutants. We infected hamsters with a pool of 42 mutants (input pool), which included control mutants with insertions in four genes previously analyzed by virulence testing (loa22, ligB, flaA1, and lic20111) and 23 mutants with disrupted signal transduction genes. We quantified the mutants in different tissues (blood, kidney and liver) at 4 days post-challenge by high-throughput sequencing and compared the frequencies of mutants recovered from tissues to their frequencies in the input pool. Control mutants that were less fit in the Tn-Seq experiment were attenuated for virulence when tested separately in the hamster model of lethal leptospirosis. Control mutants with unaltered fitness were as virulent as the wild-type strain. We identified two mutants with the transposon inserted in the same putative adenylate/guanylate cyclase gene (lic12327) that had reduced in vivo fitness in blood, kidney and liver. Both lic12327 mutants were attenuated for virulence when tested individually in hamsters. Growth of the control mutants and lic12327 mutants in culture medium were similar to that of the wild-type strain. These results demonstrate the feasibility of screening large pools of L. interrogans transposon mutants for those with altered fitness, and potentially attenuated virulence, by transposon sequencing. PMID

  11. High-Throughput Parallel Sequencing to Measure Fitness of Leptospira interrogans Transposon Insertion Mutants during Acute Infection.

    PubMed

    Lourdault, Kristel; Matsunaga, James; Haake, David A

    2016-11-01

    Pathogenic species of Leptospira are the causative agents of leptospirosis, a zoonotic disease that causes mortality and morbidity worldwide. The understanding of the virulence mechanisms of Leptospira spp is still at an early stage due to the limited number of genetic tools available for this microorganism. The development of random transposon mutagenesis in pathogenic strains a decade ago has contributed to the identification of several virulence factors. In this study, we used the transposon sequencing (Tn-Seq) technique, which combines transposon mutagenesis with massive parallel sequencing, to study the in vivo fitness of a pool of Leptospira interrogans mutants. We infected hamsters with a pool of 42 mutants (input pool), which included control mutants with insertions in four genes previously analyzed by virulence testing (loa22, ligB, flaA1, and lic20111) and 23 mutants with disrupted signal transduction genes. We quantified the mutants in different tissues (blood, kidney and liver) at 4 days post-challenge by high-throughput sequencing and compared the frequencies of mutants recovered from tissues to their frequencies in the input pool. Control mutants that were less fit in the Tn-Seq experiment were attenuated for virulence when tested separately in the hamster model of lethal leptospirosis. Control mutants with unaltered fitness were as virulent as the wild-type strain. We identified two mutants with the transposon inserted in the same putative adenylate/guanylate cyclase gene (lic12327) that had reduced in vivo fitness in blood, kidney and liver. Both lic12327 mutants were attenuated for virulence when tested individually in hamsters. Growth of the control mutants and lic12327 mutants in culture medium were similar to that of the wild-type strain. These results demonstrate the feasibility of screening large pools of L. interrogans transposon mutants for those with altered fitness, and potentially attenuated virulence, by transposon sequencing.

  12. High-Yield Synthesis of Stoichiometric Boron Nitride Nanostructures

    DOE PAGES

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

    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

  13. Optimizing the yield and selectivity of high purity nanoparticle clusters

    NASA Astrophysics Data System (ADS)

    Pease, Leonard F.

    2011-05-01

    Here we investigate the parameters that govern the yield and selectivity of small clusters composed of nanoparticles using a Monte Carlo simulation that accounts for spatial and dimensional distributions in droplet and nanoparticle density and size. Clustering nanoparticles presents a powerful paradigm with which to access properties not otherwise available using individual molecules, individual nanoparticles or bulk materials. However, the governing parameters that precisely tune the yield and selectivity of clusters fabricated via an electrospray droplet evaporation method followed by purification with differential mobility analysis (DMA) remain poorly understood. We find that the product of the electrospray droplet mean diameter to the third power and nanoparticle concentration governs the yield of individual clusters, while the ratio of the nanoparticle standard deviation to the mean diameter governs the selectivity. The resulting, easily accessible correlations may be used to minimize undesirable clustering, such as protein aggregation in the biopharmaceutical industry, and maximize the yield of a particular type of cluster for nanotechnology and energy applications.

  14. 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.

  15. 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.

  16. 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.

  17. High yield synthesis and processing of nanoscale YTZP ceramics

    NASA Astrophysics Data System (ADS)

    Szepesi, Christopher J.

    Nanomaterials are the subject of increasing interest. The expectation of new and enhanced mechanical, optical, magnetic, and electronic properties, in part due to the high concentration of interfaces and grain boundaries within the nanoscale microstructure, has initiated many studies on how to synthesize materials and process components with a final grain size below 100 nm. Several obstacles have hindered the practical application of such materials. Issues related to the production of sufficient quantities of powders composed of nanoparticles, the tendency of the nanoparticles to aggregate, the production of homogeneous green bodies, and avoiding grain growth during sintering have all been addressed with varying degrees of success. The focus of this thesis research is to address several issues inherent to the processing of nanoscale particulates, specifically in yttria tetragonal stabilized zirconia polycrystalline (YTZP) materials, for the purpose of fabricating dense, bulk components with a nano-scale microstructure. These issues include the synthesis of sufficient quantities of an appropriate material in a dispersed state, the formation of homogeneous green bodies of high green density, and design of sintering conditions that retain the fine-grain microstructure while allowing densification to near theoretical density. A recently-developed hydrothermal precipitation procedure was chosen for the production of nano-YTZP because crystalline, 8--10 nm particles of zirconia or YTZP can be produced with a flexible composition and dispersed for further processing. A wide range of characterization techniques are employed to verify particle size, phase, composition, and impurity content. Material yields are increased to 100 g of zirconia or YTZP per liter of stock solution by increasing in the reagent concentrations. Despite the increased ionic strength of the as-synthesized suspension, a laundering and dispersion procedure is described in which well

  18. High Production of 2,3-butanediol by a Mutant Strain of the Newly Isolated Klebsiella pneumoniae SRP2 with Increased Tolerance Towards Glycerol

    PubMed Central

    Rahman, Md. Shafiqur; Xu, Chunbao (Charles); Ma, Kesen; Nanda, Malaya; Qin, Wensheng

    2017-01-01

    Biodiesel, a renewable fuel produced by transesterification of animal fats and vegetable oils, generates about 10% (v/v) of crude glycerol as a core byproduct. The high volume of this non bio-degradable glycerol is becoming of a great environmental and economical concern due to its worldwide ever-growing surplus. Herein we report a high production of 2,3-butanediol (2,3-BD) from pure and biodiesel derived crude glycerol using a mutant K. pneumoniae SRM2 obtained from a newly isolated strain Klebsiella pneumoniae SRP2. The mutant strain SRM2 with standing high glycerol concentration (220 g L-1 of medium) could rapidly convert glycerol aerobically to 2,3-BD, a versatile product extensively used in chemical, pharmaceutical and fuel industries Our study revealed that an increased GDH activity led to a substantially enhanced production of 2,3-BD. The mutant strain exhibited 1.3-fold higher activity of GDH than that of parent strain (500.08 vs. 638.6 µmol min -1 mg -1 protein), yielding of 32.3 g L-1 and 77.5 g L-1 2,3-BD with glycerol in batch and fed-batch process respectively. However, in batch culture with crude glycerol, cell growth and glycerol consumption were expressively boosted, and 2,3-BD production was 27.7 g L-1 from 75.0 g/L crude glycerol. In this report, the optimal conditions for high production of 2,3-BD were defined in a completely aerobic process, and 0.59 g g-1 product yield of 2,3-BD was attained by the mutated strain K. pneumoniae SRM2, which is the highest amount obtained from batch biotransformation process of glycerol metabolism till today. These results indicated that our newly developed mutant can tolerate high concentration of glycerol, have a high glycerol utilization rate, and high product yield of 2,3-BD. It is demonstrated that the mutant strain K. pneumoniae SRM2 has an ability to produce fewer co-products at trace concentrations at higher glycerol concentrations, and could be a potential candidate for 2,3-DB production in an industrial

  19. 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.

  20. 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.

  1. Yield-enhanced routing for high-performance VLSI designs

    NASA Astrophysics Data System (ADS)

    Venkataraman, Arunshankar; Chen, Howard H.; Koren, Israel

    1997-09-01

    It is widely recognized that interconnects will be the main bottleneck in enhancing the performance of future deep sub-micron VLSI designs. Interconnects do not "scale" well with decreasing feature sizes and therefore dominate the delays in the integrated circuit. In addition to RC delays, crosstalk noise also contributes significantly to the delays experienced by a signal. Interconnects are more susceptible to manufacturing defects and therefore affect the product yields significantly. Recently, several channel-routing based solutions have been proposed to minimize crosstalk noise and also enhance yield of the routing. While these approaches are effective, they do not provide maximum benefits as they are either constrained by a particular design methodology or are post-routing steps which have limited scope for significant improvement. Also, design for manufacturabiity objectives have not been fully exploited by VLSI CAD tools as they do not integrate seamlessly into the conventional design flow and the added overheads make it less attractive. In this paper, we propose a modified routing algorithm that maximizes yield and reduces crosstalk noise while using minimal area for the routing. The yield enhancement objective has been integrated into the routing phase as a preferred constraint (a constraint that will be satisfied only if the primary constraints of minimal area and wire length have been satisfied) and fits well into the conventional design flow. This enables the router to produce an output which provides maximum achievable critical area reduction for the given routing solution. Post-routing layout modification is also done with the objective of minimizing the interaction area between the interconnects by exploiting the gridless property of the router. The above algorithm is incorporated into GLITTER (the gridless, variable width channel router), and the results on channel-routing benchmarks are presented. These results show a significant reduction in the

  2. Butanol production by a Clostridium beijerinckii mutant with high ferulic acid tolerance.

    PubMed

    Liu, Jun; Guo, Ting; Wang, Dong; Xu, Jiahui; Ying, Hanjie

    2016-09-01

    A mutant strain of Clostridium beijerinckii, with high tolerance to ferulic acid, was generated using atmospheric pressure glow discharge and high-throughput screening of C. beijerinckii NCIMB 8052. The mutant strain M11 produced 7.24 g/L of butanol when grown in P2 medium containing 30 g/L of glucose and 0.5 g/L of ferulic acid, which is comparable to the production from non-ferulic acid cultures (8.11 g/L of butanol). When 0.8 g/L of ferulic acid was introduced into the P2 medium, C. beijerinckii M11 grew well and produced 4.91 g/L of butanol. Both cell growth and butanol production of C. beijerinckii M11 were seriously inhibited when 0.9 g/L of ferulic acid was added into the P2 medium. Furthermore, C. beijerinckii M11 could produce 6.13 g/L of butanol using non-detoxified hemicellulosic hydrolysate from diluted sulfuric acid-treated corn fiber (SAHHC) as the carbon source. These results demonstrate that C. beijerinckii M11 has a high ferulic acid tolerance and is able to use non-detoxified SAHHC for butanol production.

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

    PubMed Central

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

    2005-01-01

    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. PMID:16170329

  4. Insights into the conformational equilibria of maltose-binding protein by analysis of high affinity mutants.

    PubMed

    Telmer, Patrick G; Shilton, Brian H

    2003-09-05

    The affinity of maltose-binding protein (MBP) for maltose and related carbohydrates was greatly increased by removal of groups in the interface opposite the ligand binding cleft. The wild-type protein has a KD of 1200 nM for maltose; mutation of residues Met-321 and Gln-325, both to alanine, resulted in a KD for maltose of 70 nM; deletion of 4 residues, Glu-172, Asn-173, Lys-175, and Tyr-176, which are part of a poorly ordered loop, results in a KD for maltose of 110 nM. Combining the mutations yields an increased affinity for maltodextrins and a KD of 6 nM for maltotriose. Comparison of ligand binding by the mutants, using surface plasmon resonance spectroscopy, indicates that decreases in the off-rate are responsible for the increased affinity. Small-angle x-ray scattering was used to demonstrate that the mutations do not significantly affect the solution conformation of MBP in either the presence or absence of maltose. The crystal structures of selected mutants showed that the mutations do not cause significant structural changes in either the closed or open conformation of MBP. These studies show that interactions in the interface opposite the ligand binding cleft, which we term the "balancing interface," are responsible for modulating the affinity of MBP for its ligand. Our results are consistent with a model in which the ligand-bound protein alternates between the closed and open conformations, and removal of interactions in the balancing interface decreases the stability of the open conformation, without affecting the closed conformation.

  5. 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.

  6. Exploring the elephant: histopathology in high-throughput phenotyping of mutant mice.

    PubMed

    Schofield, Paul N; Vogel, Peter; Gkoutos, Georgios V; Sundberg, John P

    2012-01-01

    Recent advances in gene knockout techniques and the in vivo analysis of mutant mice, together with the advent of large-scale projects for systematic mouse mutagenesis and genome-wide phenotyping, have allowed the creation of platforms for the most complete and systematic analysis of gene function ever undertaken in a vertebrate. The development of high-throughput phenotyping pipelines for these and other large-scale projects allows investigators to search and integrate large amounts of directly comparable phenotype data from many mutants, on a genomic scale, to help develop and test new hypotheses about the origins of disease and the normal functions of genes in the organism. Histopathology has a venerable history in the understanding of the pathobiology of human and animal disease, and presents complementary advantages and challenges to in vivo phenotyping. In this review, we present evidence for the unique contribution that histopathology can make to a large-scale phenotyping effort, using examples from past and current programmes at Lexicon Pharmaceuticals and The Jackson Laboratory, and critically assess the role of histopathology analysis in high-throughput phenotyping pipelines.

  7. Prediction of yield stress in highly irradiated ferritic steels

    NASA Astrophysics Data System (ADS)

    Windsor, Colin G.; Cottrell, Geoff; Kemp, Richard

    2008-03-01

    The design of any fusion power plant requires information on the irradiation hardening of low-activation ferritic/martensitic steels beyond the range of most present measurements. Neural networks have been used by Kemp et al (J. Nucl. Mater. 348 311-28) to model the yield stress of some 1811 irradiated alloys. The same dataset has been used in this study, but has been divided into a training set containing the majority of the dataset with low irradiation levels, and a test set which contains just those alloys which have been irradiated above a given level. For example some 4.5% of the alloys were irradiated above 30 displacements per atom. For this 'prediction' problem it is found that simpler networks with fewer inputs are advantageous. By using target-driven dimensionality reduction, linear combinations of the atomic inputs reduce the test residual below that achievable by adding inputs from single atoms. It is postulated that these combinations represent 'mechanisms' for the prediction of irradiated yield stress.

  8. 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.

  9. 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.

  10. Modifications of und rbcS in und Synechococcus sp. PCC7942 resulted in high CO sub 2 requiring mutants

    SciTech Connect

    Schwarz, R.; Liehman-Hurwitz, J.; Martinez, F.; Reinhold, L.; Kaplan, A. )

    1990-05-01

    High CO{sub 2} requiring mutants (HCR), capable of growing in the presence of kanamycin, were obtained following transformation with a construct containing {und nptII} in the coding region of rbcS (encoding for the small subunit (SS) of Rubisco). The apparent photosynthetic affinity for inorganic carbon was very low in mutant EK6 whereas in mutant JR12 it was only slightly lower than in high-CO{sub 2}-grown wild type. Southern analysis indicated double crossing over in the case of EK6. In mutant JR12 Southern analysis revealed some rearrangement and a deletion of a region downstream fro {und rbcS}. Western analysis demonstrated normal SS in JR12 but a larger (by about 3 kD) SS in EK6. Sequence analysis indicated that the latter was the result of fusion between {und rbcS} and the flanking region of {und nptII}.

  11. 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.

  12. Yield mapping of high-biomass sorghum with aerial imagery

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 high biomass...

  13. 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

  14. 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...

  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. A highly digestible sorghum mutant cultivar exhibits a unique folded structure of endosperm protein bodies

    PubMed Central

    Oria, Maria P.; Hamaker, Bruce R.; Axtell, John D.; Huang, Chia-Ping

    2000-01-01

    The endosperm of a sorghum mutant cultivar, with high in vitro uncooked and cooked protein digestibilities, was examined by transmission electron microscopy and α-, β-, and γ-kafirins (storage proteins) were localized within its protein bodies. Transmission electron microscopy micrographs revealed that these protein bodies had a unique microstructure related to high protein digestibility. They were irregular in shape and had numerous invaginations, often reaching to the central area of the protein body. Protein bodies from normal cultivars, such as P721N studied here, with much lower uncooked and cooked digestibilities are spherical and contain no invaginations. Immunocytochemistry results showed that the relative location of α- and β-kafirins within the protein bodies of the highly digestible genotype were similar to the normal cultivar, P721N. γ-Kafirin, however, was concentrated in dark-staining regions at the base of the folds instead of at the protein body periphery, as is typical of normal cultivars. The resulting easy accessibility of digestive enzymes to α-kafirin, the major storage protein, in addition to the increased surface area of the protein bodies of the highly digestible cultivar appear to account for its high in vitro protein digestibility. PMID:10792028

  17. 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...

  18. Controllable preparation of high-yield magnetic polymer latex.

    PubMed

    Wu, Chun-Chao; Kong, Xiang-Ming; Yang, Hai-Long

    2011-09-01

    In order to overcome the low conversion and complex post-treatment, four different polymerization procedures were adopted to prepare the magnetic polymer latexes. The results clearly show that the strategy using magnetic emulsion template-dosage is the most effective and feasible. Based on the optimized procedure, various factors including the type of initiators such as oil soluble initiator, water soluble initiator, redox initiator system, crosslinking agent, functional monomers etc. were systematically studied. Magnetic polymer latex with high monomer conversion of 83% and high magnet content of 31.8% was successfully obtained. Besides, core-shell structured magnetic polymer latex with good film forming property was also prepared, which is promising for potential applications such as magnetic coatings and modification of cementitious materials with controlled polymer location.

  19. High yield electroblotting onto polyvinylidene difluoride membranes from polyacrylamide gels.

    PubMed

    Mozdzanowski, J; Hembach, P; Speicher, D W

    1992-01-01

    Optimal conditions of electroblotting that led to high protein recovery on polyvinylidene difluoride (PVDF) membranes were determined for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). SDS concentrations in the gel and transfer buffer were found to be the most important factors affecting the amount of protein recovered on the PVDF membrane. The largest loss occurred during the first 10-30 min of transfer due to the relatively high initial SDS concentration in the gel. During this initial stage of transfer, most of the protein passed through the primary membrane and was partially retained on secondary and tertiary membranes. The value of presoaking gels prior to transfer to reduce the amount of SDS was evaluated by quantitating free SDS densitometrically and by correlating the reduced SDS concentration with increased electroblotting efficiency from presoaked gels. Transfer time was evaluated and no "overtransfer" was found even after very long transfer times. These results clearly indicate that proteins electroblotted onto PVDF membranes were tightly bound and could not be released by extending the transfer time. The effects of methanol and SDS concentrations on protein adsorption from solution to PVDF were also determined quantitatively. The results of this study strongly suggest that proteins fully saturated with SDS cannot bind efficiently to PVDF membranes. Since SDS is necessary for high protein mobility, the challenge in efficient electroblotting is to maintain an optimal SDS concentration which is high enough to permit effective removal from the gel and low enough to permit effective binding to the PVDF membrane. For 1.5 mm thick gels containing 0.2% SDS, presoaking the gel for 15-20 min in transfer buffer with 10% methanol prior to electroblotting provided the best recovery on the primary membrane.

  20. 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.

  1. 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.

  2. 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.

  3. Abnormal Mammary Adipose Tissue Environment of Brca1 Mutant Mice Show a Persistent Deposition of Highly Vascularized Multilocular Adipocytes.

    PubMed

    Jones, Laundette P; Buelto, Destiney; Tago, Elaine; Owusu-Boaitey, Kwadwo E

    2011-12-08

    A major challenge to breast cancer research is the identification of alterations in the architecture and composition of the breast that are associated with breast cancer progression. The aim of the present investigation was to characterize the mammary adipose phenotype from Brca1 mutant mice in the expectation that this would shed light on the role of the mammary tissue environment in the early stages of breast tumorigenesis. We observed that histological sections of mammary tissue from adult Brca1 mutant mice abnormally display small, multilocular adipocytes that are reminiscent of brown adipose tissue (BAT) as compared to wildtype mice. Using a marker for BAT, the uncoupling protein 1 (UCP1), we demonstrated that these multilocular adipose regions in Brca1 mutant mice stain positive for UCP1. Transcriptionally, UCP1 mRNA levels in the Brca1 mutant mice were elevated greater than 50-fold compared to age-matched mammary glands from wildtype mice. Indeed, BAT has characteristics that are favorable for tumor growth, including high vascularity. Therefore, we also demonstrated that the multilocular brown adipose phenotype in the mammary fat pad of Brca1 mutant mice displayed regions of increased vascularity as evidenced by a significant increase in the protein expression of CD31, a marker for angiogenesis. This Brca1 mutant mouse model should provide a physiologically relevant context to determine whether brown adipose tissue can play a role in breast cancer development.

  4. High yield NMR, FY86-2: Quarterly report

    SciTech Connect

    Smith, R.E.

    1986-11-01

    Several analytical methods were investigated this quarter. The previously developed method for determining the percent vinyl and phenyl in silicones has been modified, because it was discovered that the silicone gums are not perfectly homogeneous. Standards having a known vinyl and phenyl content were analyzed to confirm the accuracy of the NMR method. In addition, a high temperature carbon-13 NMR analysis of high strength, low density polymethylpentene (TPX) polymer was performed. A solder flux used on several programs was analyzed and the activator was identified as the hydrochloric acid (HCl) salt of glutamic acid. The reactions that occur in the model epoxy system, phenyl glycidyl ether plus dicyandiamide (DICY), plus boron trifluoride monoethylamine (BF/sub 3/-MEA) were investigated. The solvent carriers in two epoxies were identified. An alkaline cleaner was analyzed and a method was developed to estimate changes in detergent concentration. Methods based on carbon-13 NMR were developed to test the purity of organic compounds. 7 refs., 10 figs., 5 tabs.

  5. Simple, high-yield synthesis of polyhedral carborane amino acids

    SciTech Connect

    Kahl, S.B.; Kasar, R.A.

    1996-02-07

    Boron neutron capture therapy (BNCT) is a form of binary cancer therapy that offers the potential of delivering spatially selective, high linear energy transfer radiation to the target cells while sparing surrounding normal tissue. We have demonstarted a versatile, general method for the conversion of o- ,m-, and p-carborane to their corresponding Boc-protected amino acids. Heterobifunctional polyhedral carboranes are exceedingly rare in the literature, and the amino acids prepared by this general method may prove to be valuable synthons for use in the synthesis of tumor-seeking compounds for BNCT or PDT. Morever, these conformationally constrained amino acids should be particularly interesting for use in peptide synthesis. The dihedral angle between the carbon atoms of these polyhedra increases in the order 60{degree} (ortho), 110{degree} (meta), and 180{degree} (para), allowing the peptide chemist to select a desired conformation. 11 refs.

  6. A high-yield ionic liquid-promoted synthesis of boron nitride nanosheets by direct exfoliation.

    PubMed

    Morishita, Takuya; Okamoto, Hirotaka; Katagiri, Yoshihide; Matsushita, Mitsumasa; Fukumori, Kenzo

    2015-08-04

    Boron nitride nanosheets (BNNSs) with micron-sized edges were prepared in high yields by direct exfoliation of bulk hexagonal boron nitrides using ionic liquids (ILs). The ILs strongly attached onto BNNS surfaces, and dramatically enhanced the exfoliation, giving highly concentrated BNNS dispersions (∼1.9 mg mL(-1)) and yields reaching ∼50%.

  7. A natural variant of NAL1, selected in high-yield rice breeding programs, pleiotropically increases photosynthesis rate

    PubMed Central

    Takai, Toshiyuki; Adachi, Shunsuke; Taguchi-Shiobara, Fumio; Sanoh-Arai, Yumiko; Iwasawa, Norio; Yoshinaga, Satoshi; Hirose, Sakiko; Taniguchi, Yojiro; Yamanouchi, Utako; Wu, Jianzhong; Matsumoto, Takashi; Sugimoto, Kazuhiko; Kondo, Katsuhiko; Ikka, Takashi; Ando, Tsuyu; Kono, Izumi; Ito, Sachie; Shomura, Ayahiko; Ookawa, Taiichiro; Hirasawa, Tadashi; Yano, Masahiro; Kondo, Motohiko; Yamamoto, Toshio

    2013-01-01

    Improvement of leaf photosynthesis is an important strategy for greater crop productivity. Here we show that the quantitative trait locus GPS (GREEN FOR PHOTOSYNTHESIS) in rice (Oryza sativa L.) controls photosynthesis rate by regulating carboxylation efficiency. Map-based cloning revealed that GPS is identical to NAL1 (NARROW LEAF1), a gene previously reported to control lateral leaf growth. The high-photosynthesis allele of GPS was found to be a partial loss-of-function allele of NAL1. This allele increased mesophyll cell number between vascular bundles, which led to thickened leaves, and it pleiotropically enhanced photosynthesis rate without the detrimental side effects observed in previously identified nal1 mutants, such as dwarf plant stature. Furthermore, pedigree analysis suggested that rice breeders have repeatedly selected the high-photosynthesis allele in high-yield breeding programs. The identification and utilization of NAL1 (GPS) can enhance future high-yield breeding and provides a new strategy for increasing rice productivity. PMID:23985993

  8. High-Throughput Robotically Assisted Isolation of Temperature-sensitive Lethal Mutants in Chlamydomonas reinhardtii

    PubMed Central

    Breker, Michal; Lieberman, Kristi; Tulin, Frej; Cross, Frederick R.

    2016-01-01

    Systematic identification and characterization of genetic perturbations have proven useful to decipher gene function and cellular pathways. However, the conventional approaches of permanent gene deletion cannot be applied to essential genes. We have pioneered a unique collection of ~70 temperature-sensitive (ts) lethal mutants for studying cell cycle regulation in the unicellular green algae Chlamydomonas reinhardtii1. These mutations identify essential genes, and the ts alleles can be conditionally inactivated by temperature shift, providing valuable tools to identify and analyze essential functions. Mutant collections are much more valuable if they are close to comprehensive, since scattershot collections can miss important components. However, this requires the efficient collection of a large number of mutants, especially in a wide-target screen. Here, we describe a robotics-based pipeline for generating ts lethal mutants and analyzing their phenotype in Chlamydomonas. This technique can be applied to any microorganism that grows on agar. We have collected over 3000 ts mutants, probably including mutations in most or all cell-essential pathways, including about 200 new candidate cell cycle mutations. Subsequent molecular and cellular characterization of these mutants should provide new insights in plant cell biology; a comprehensive mutant collection is an essential prerequisite to ensure coverage of a broad range of biological pathways. These methods are integrated with downstream genetics and bioinformatics procedures for efficient mapping and identification of the causative mutations that are beyond the scope of this manuscript. PMID:28060315

  9. Molecular characterization of two high-palmitic-acid mutant loci induced by X-ray irradiation in soybean.

    PubMed

    Anai, Toyoaki; Hoshino, Tomoki; Imai, Naoko; Takagi, Yutaka

    2012-01-01

    Palmitic acid is the most abundant (approx. 11% of total fatty acids) saturated fatty acid in conventional soybean seed oil. Increasing the saturated acid content of soybean oil improves its oxidative stability and plasticity. We have developed three soybean mutants with high palmitic acid content by X-ray irradiation. In this study, we successfully identified the mutated sites of two of these high-palmitic-acid mutants, J10 and M22. PCR-based mutant analysis revealed that J10 has a 206,203-bp-long deletion that includes the GmKASIIA gene and 16 other predicted genes, and M22 has a 26-bp-long deletion in the sixth intron of GmKASIIB. The small deletion in M22 causes mis-splicing of GmKASIIB transcripts, which should result in nonfunctional products. In addition, we designed co-dominant marker sets for these mutant alleles and confirmed the association of genotypes and palmitic acid contents in F(2) seeds of J10 X M22. This information will be useful in breeding programs to develop novel soybean cultivars with improved palmitic acid content. However, in the third mutant, KK7, we found no polymorphism in either GmKASIIA or GmKASIIB, which suggests that several unknown genes in addition to GmKASIIA and GmKASIIB may be involved in elevating the palmitic acid content of soybean seed oil.

  10. 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.

  11. Automated Purification of Recombinant Proteins: Combining High-throughput with High Yield

    SciTech Connect

    Lin, Chiann Tso; Moore, Priscilla A.; Auberry, Deanna L.; Landorf , Elizabeth V.; Peppler , Teresa; Victry, Kristin D.; Collart, Frank R.; Kery, Vladimir

    2006-05-01

    Protein crystallography, mapping protein interactions and other approaches of current functional genomics require not only purifying large numbers of proteins but also obtaining sufficient yield and homogeneity for downstream high-throughput applications. There is a need for the development of robust automated high-throughput protein expression and purification processes to meet these requirements. We developed and compared two alternative workflows for automated purification of recombinant proteins based on expression of bacterial genes in Escherichia coli: First - a filtration separation protocol based on expression of 800 ml E. coli cultures followed by filtration purification using Ni2+-NTATM Agarose (Qiagen). Second - a smaller scale magnetic separation method based on expression in 25 ml cultures of E.coli followed by 96-well purification on MagneHisTM Ni2+ Agarose (Promega). Both workflows provided comparable average yields of proteins about 8 ug of purified protein per unit of OD at 600 nm of bacterial culture. We discuss advantages and limitations of the automated workflows that can provide proteins more than 90 % pure in the range of 100 ug – 45 mg per purification run as well as strategies for optimization of these protocols.

  12. 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.

  13. 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

  14. Use of a high-throughput screen to identify Leptospira mutants unable to colonize the carrier host or cause disease in the acute model of infection.

    PubMed

    Marcsisin, Renee A; Bartpho, Thanatchaporn; Bulach, Dieter M; Srikram, Amporn; Sermswan, Rasana W; Adler, Ben; Murray, Gerald L

    2013-10-01

    The molecular basis for leptospirosis infection and colonization remains poorly understood, with no efficient methods available for screening libraries of mutants for attenuation. We analysed the attenuation of leptospiral transposon mutants in vivo using a high-throughput method by infecting animals with pooled sets of transposon mutants. A total of 95 mutants was analysed by this method in the hamster model of acute infection, and one mutant was identified as attenuated (M1233, lb058 mutant). All virulence factors identified in Leptospira to date have been characterized in the acute model of infection, neglecting the carrier host. To address this, a BALB/c mouse colonization model was established. The lb058 mutant and two mutants defective in LPS synthesis were colonization deficient in the mouse model. By applying the high-throughput screening method, a further five colonization-deficient mutants were identified for the mouse model; these included two mutants in genes encoding proteins with a predicted role in iron uptake (LB191/HbpA and LB194). Two attenuated mutants had transposon insertions in either la0589 or la2786 (encoding proteins of unknown function). The final attenuated mutant had an unexpected deletion of genes la0969-la0975 at the point of transposon insertion. This is the first description of defined, colonization-deficient mutants in a carrier host for Leptospira. These mutants were either not attenuated or only weakly attenuated in the hamster model of acute leptospirosis, thus illustrating that different factors that may be required in the carrier and acute models of leptospiral infection. High-throughput screening can reduce the number of animals used in virulence studies and increase the capacity to screen mutants for attenuation, thereby enhancing the likelihood of detecting unique virulence factors. A comparison of virulence factors required in the carrier and acute models of infection will help to unravel colonization and dissemination

  15. Head rice yield response to low and high drying and tempering conditions

    NASA Astrophysics Data System (ADS)

    Sadeghi, M.; Nasrnia, E.; Masoumi, A. A.; Hemmat, A.

    2013-03-01

    The influence of drying and tempering conditions on head rice yield of long- and medium-grain rough rice varieties was investigated. The head rice yield values for the medium-grain variety at high drying conditions and 1.5 and 3% points moisture content removal were even significantly higher than the corresponding values for the long-grain variety at low drying conditions. 1.5% points moisture content removal yielded the least damage to the rough rice. Under low drying conditions, tempering had no meaningful effect on head rice yield for all drying durations. For both varieties, using high drying conditions with 6% points moisture content removal at first drying stage associated with 120 min tempering duration could be suggested as a rapid and energysaving operation to achieve high head rice yield values.

  16. Relationships between fruit exocarp antioxidants in the tomato (Lycopersicon esculentum) high pigment-1 mutant during development.

    PubMed

    Andrews, Preston K.; Fahy, Deirdre A.; Foyer, Christine H.

    2004-04-01

    Development-dependent changes in fruit antioxidants were examined in the exocarp (epidermal and hypodermal tissues) of the monogenic recessive tomato (Lycopersicon esculentum L.) mutant high pigment (hp-1) and its wild-type parent 'Rutgers' grown under non-stress conditions in a greenhouse. The hp-1 mutant was chosen for this study because the reportedly higher lycopene and ascorbic acid (AsA) contents of the fruit may alter its tolerance to photooxidative stress. Throughout most of fruit development, reduced AsA concentrations in the exocarp of hp-1 were 1.5 to 2.0 times higher than in 'Rutgers', but total glutathione concentrations were similar in both genotypes. Only in ripe red fruit were reduced AsA and total glutathione concentrations lower in hp-1 than in 'Rutgers'. The redox ratios (reduced : reduced + oxidized) of AsA in hp-1 and 'Rutgers' exocarps were similar and usually > 0.9, however, the redox ratio of glutathione was lower in hp-1 than in 'Rutgers' throughout development. Lycopene concentrations in ripe red fruit were about 5 times higher in hp-1 than in 'Rutgers'. Large increases in the specific enzyme activities of superoxide dismutase (EC 1.15.1.1), ascorbate peroxidase (EC 1.11.1.11), and monodehydroascorbate reductase (MDHAR; EC 1.6.5.4) occurred during fruit development in both genotypes, with an inverse relationship between the activities of these enzymes and chlorophyll content. Glutathione reductase (EC 1.6.4.2) and MDHAR-specific activities were higher in hp-1 than 'Rutgers' only at the later stages of fruit development. Dehydroascorbate reductase (EC 1.8.5.1) activities, however, were usually higher in 'Rugters' than in hp-1. Catalase (CAT, EC 1.11.1.6) activities increased with fruit development until the fruit were orange/light red, when CAT was higher in 'Rutgers' than in hp-1, but then declined in the ripe red fruit of both genotypes. These results suggest that elevated AsA in the exocarp of hp-1 fruit early in fruit development may

  17. A proteomic study of rice cultivar TNG67 and its high aroma mutant SA0420.

    PubMed

    Lin, Da-Gin; Chou, Szu-Yi; Wang, Arthur Z; Wang, Yi-Wen; Kuo, Shu-Ming; Lai, Chien-Chen; Chen, Liang-Jwu; Wang, Chang-Sheng

    2014-01-01

    Fragrance is a very important economic trait for rice cultivars. To identify the aroma genes in rice, we performed a proteomics analysis of aroma-related proteins between Tainung 67 (TNG67) and its high aroma mutant SA0420. Seventeen of the differentially identified proteins were close related with the aroma phenotype of SA0420. Among them, 9 were found in leaves and 8 were found in grains. One protein (L3) was identified as the chloroplastic glyceraldehyde-3-phosphate dehydrogenase B (OsGAPDHB) which was less abundant in SA0420 than TNG67. Sequence analysis demonstrated that this protein in SA0420 carries a P425S mutation in the C-terminal extension domain, which might hinder the formation of holoenzyme, thereby changing the profile of aroma compounds. The protein profile of OsGAPDHB showed only a weak correlation to its transcription profile. This result indicated that the reduction of OsGAPDHB in SA0420 is regulated by post-translational processes and can only be analyzed by proteomics approach. Transgenic lines suppressing OsGAPDHB through RNAi harbored more fragrance than TNG67 but less than SA0420. With betaine-aldehyde dehydrogenase as the only fragrance gene identified in rice to date, OsGAPDHB may serve as the second protein known to contribute to the aroma phenotype.

  18. 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...

  19. 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...

  20. 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.

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

    PubMed

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

    2016-07-01

    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.

  2. 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

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

    PubMed

    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.

  4. Simplifying multidimensional fermentation dataset analysis and visualization: One step closer to capturing high-quality mutant strains

    PubMed Central

    Zhou, Xiang; Xu, Dan; Jiang, Ting-Ting

    2017-01-01

    In this study, we analyzed mutants of Clostridium acetobutylicum, an organism used in a broad range of industrial processes related to biofuel production, to facilitate future studies of bioreactor and bioprocess design and scale-up, which are very important research projects for industrial microbiology applications. To accomplish this, we generated 329 mutant strains and applied principal component analysis (PCA) to fermentation data gathered from these strains to identify a core set of independent features for comparison. By doing so, we were able to explain the differences in the mutant strains’ fermentation expression states and simplify the analysis and visualization of the multidimensional datasets related to the strains. Our study has produced a high-efficiency PCA application based on a data analytics tool that is designed to visualize screening results and to support several hundred sets of data on fermentation interactions to assist researchers in more precisely screening and capturing high-quality mutant strains. More importantly, although this study focused on the use of PCA in microbial fermentation engineering, its results are broadly applicable. PMID:28045110

  5. High-throughput genotyping of CRISPR/Cas9-mediated mutants using fluorescent PCR-capillary gel electrophoresis

    PubMed Central

    Ramlee, Muhammad Khairul; Yan, Tingdong; Cheung, Alice M. S.; Chuah, Charles T. H.; Li, Shang

    2015-01-01

    Recent advances in the engineering of sequence-specific synthetic nucleases provide enormous opportunities for genetic manipulation of gene expression in order to study their cellular function in vivo. However, current genotyping methods to detect these programmable nuclease-induced insertion/deletion (indel) mutations in targeted human cells are not compatible for high-throughput screening of knockout clones due to inherent limitations and high cost. Here, we describe an efficient method of genotyping clonal CRISPR/Cas9-mediated mutants in a high-throughput manner involving the use of a direct lysis buffer to extract crude genomic DNA straight from cells in culture, and fluorescent PCR coupled with capillary gel electrophoresis. This technique also allows for genotyping of multiplexed gene targeting in a single clone. Overall, this time- and cost-saving technique is able to circumvent the limitations of current genotyping methods and support high-throughput screening of nuclease-induced mutants. PMID:26498861

  6. High Resolution Melt (HRM) analysis is an efficient tool to genotype EMS mutants in complex crop genomes

    PubMed Central

    2011-01-01

    Background Targeted Induced Loci Lesions IN Genomes (TILLING) is increasingly being used to generate and identify mutations in target genes of crop genomes. TILLING populations of several thousand lines have been generated in a number of crop species including Brassica rapa. Genetic analysis of mutants identified by TILLING requires an efficient, high-throughput and cost effective genotyping method to track the mutations through numerous generations. High resolution melt (HRM) analysis has been used in a number of systems to identify single nucleotide polymorphisms (SNPs) and insertion/deletions (IN/DELs) enabling the genotyping of different types of samples. HRM is ideally suited to high-throughput genotyping of multiple TILLING mutants in complex crop genomes. To date it has been used to identify mutants and genotype single mutations. The aim of this study was to determine if HRM can facilitate downstream analysis of multiple mutant lines identified by TILLING in order to characterise allelic series of EMS induced mutations in target genes across a number of generations in complex crop genomes. Results We demonstrate that HRM can be used to genotype allelic series of mutations in two genes, BraA.CAX1a and BraA.MET1.a in Brassica rapa. We analysed 12 mutations in BraA.CAX1.a and five in BraA.MET1.a over two generations including a back-cross to the wild-type. Using a commercially available HRM kit and the Lightscanner™ system we were able to detect mutations in heterozygous and homozygous states for both genes. Conclusions Using HRM genotyping on TILLING derived mutants, it is possible to generate an allelic series of mutations within multiple target genes rapidly. Lines suitable for phenotypic analysis can be isolated approximately 8-9 months (3 generations) from receiving M3 seed of Brassica rapa from the RevGenUK TILLING service. PMID:22152063

  7. Fed-Batch Enzymatic Saccharification of High Solids Pretreated Lignocellulose for Obtaining High Titers and High Yields of Glucose.

    PubMed

    Jung, Young Hoon; Park, Hyun Min; Kim, Dong Hyun; Yang, Jungwoo; Kim, Kyoung Heon

    2017-01-11

    To reduce the distillation costs of cellulosic ethanol, it is necessary to produce high sugar titers in the enzymatic saccharification step. To obtain high sugar titers, high biomass loadings of lignocellulose are necessary. In this study, to overcome the low saccharification yields and the low operability of high biomass loadings, a fed-batch saccharification process was developed using an enzyme reactor that was designed and built in-house. After optimizing the cellulase and biomass feeding profiles and the agitation speed, 132.6 g/L glucose and 76.0% theoretical maximum glucose were obtained from the 60 h saccharification of maleic acid-pretreated rice straw at a 30% (w/v) solids loading with 15 filter paper units (FPU) of Cellic CTec2/g glucan. This study demonstrated that through the proper optimization of fed-batch saccharification, both high sugar titers and high saccharification yields are possible, even with using the high solids loading (i.e., ≥30%) with the moderate enzyme loading (i.e., <15 FPU/g glucan). These results could be contributed to improving economic feasibility of the high solids saccharification process in cellulosic fuel and chemical production.

  8. 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.

  9. 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.

  10. Analysis on differential expressed genes of ovarian tissue between high- and low-yield laying hen.

    PubMed

    Chen, Wei; Song, Ling-Jun; Zeng, Yong-Qing; Yang, Yun; Wang, Hui

    2013-01-01

    In order to elucidate molecular genetic mechanism of laying hen reproduction at the transcriptional level and the structure of significantly differential genes, the mRNA differential display and reverse northern dot-blot were used to detect the differential expression of genes in the ovary tissue of low-yield laying hens and high-yield laying hens in the present study. Sixteen 32-week-old CAU-pink laying hens divided into two groups were used and the laying performance was measured. The results showed that only the egg numbers were significantly different between the two groups; and from 15 primer pairs, a total of 336 bands were displayed of which 59 cDNA bands were found to be differentially expressed in both high-yield and low-yield laying hen. The sequence analysis indicated that the expression of such bands as H-AP5, H-P5, and H-P4 was significantly potentiated in high-yield laying hen using primer pairs AP5/HT11G, P5/HT11G and P4/HT11G and these transcripts had high homology (98%) to HoxDb, HoxCa, and HoxBa, respectively. The differentially expressed gene fragments may be relevant to the progression of the high-yield hens to the egg-laying stage. And further study is required to elucidate the molecular function to improve the productivity of laying hens.

  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. 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...

  13. A modified method for calculating practical ethanol yield at high lignocellulosic solids content and high ethanol titer.

    PubMed

    Zhang, Jian; Bao, Jie

    2012-07-01

    A modified method for calculating practical ethanol yield in the simultaneous saccharification and fermentation (SSF) at high lignocellulosic solids content and high ethanol titer is proposed considering the liquid volume change caused by high titer ethanol generation and the water consumed during cellulose degradation. This modified method was applied to determine the practical ethanol yields of several practical SSF operations and the results compared to those using the conventional method. The results show that the liquid volume increase with ethanol formation during SSF was approximately five times greater than the volume decrease duo to water consumption during cellulose degradation. Furthermore, the practical ethanol yields calculating using traditional method were underestimated and the underestimated errors increased with the increasing ethanol titer. The present work may provide a convenient and accurate method for calculating practical ethanol yield in a high solids and high ethanol titer SSF systems.

  14. 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

  15. 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

  16. 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.

  17. Immunological profiling of molecularly classified high-risk endometrial cancers identifies POLE-mutant and microsatellite unstable carcinomas as candidates for checkpoint inhibition.

    PubMed

    Eggink, Florine A; Van Gool, Inge C; Leary, Alexandra; Pollock, Pamela M; Crosbie, Emma J; Mileshkin, Linda; Jordanova, Ekaterina S; Adam, Julien; Freeman-Mills, Luke; Church, David N; Creutzberg, Carien L; De Bruyn, Marco; Nijman, Hans W; Bosse, Tjalling

    2017-01-01

    High-risk endometrial cancer (EC) is an aggressive disease for which new therapeutic options are needed. Aims of this study were to validate the enhanced immune response in highly mutated ECs and to explore immune profiles in other EC subgroups. We evaluated immune infiltration in 116 high-risk ECs from the TransPORTEC consortium, previously classified into four molecular subtypes: (i) ultramutated POLE exonuclease domain-mutant ECs (POLE-mutant); (ii) hypermutated microsatellite unstable (MSI); (iii) p53-mutant; and (iv) no specific molecular profile (NSMP). Within The Cancer Genome Atlas (TCGA) EC cohort, significantly higher numbers of predicted neoantigens were demonstrated in POLE-mutant and MSI tumors compared with NSMP and p53-mutants. This was reflected by enhanced immune expression and infiltration in POLE-mutant and MSI tumors in both the TCGA cohort (mRNA expression) and the TransPORTEC cohort (immunohistochemistry) with high infiltration of CD8(+) (90% and 69%), PD-1(+) (73% and 69%) and PD-L1(+) immune cells (100% and 71%). Notably, a subset of p53-mutant and NSMP cancers was characterized by signs of an antitumor immune response (43% and 31% of tumors with high infiltration of CD8(+) cells, respectively), despite a low number of predicted neoantigens. In conclusion, the presence of enhanced immune infiltration, particularly high numbers of PD-1 and PD-L1 positive cells, in highly mutated, neoantigen-rich POLE-mutant and MSI endometrial tumors suggests sensitivity to immune checkpoint inhibitors.

  18. Single cell protein production from yacon extract using a highly thermosensitive and permeable mutant of the marine yeast Cryptococcus aureus G7a and its nutritive analysis.

    PubMed

    Zhao, Chun-Hai; Zhang, Tong; Chi, Zhen-Ming; Chi, Zhe; Li, Jing; Wang, Xiang-Hong

    2010-06-01

    The intracellular protein in the highly thermosensitive and permeable mutant can be easily released when they are incubated both in the low-osmolarity water and at the non-permissive temperature (usually 37 degrees C). After the mutant was grown in the yacon extract for 45 h, the crude protein content in the highly thermosensitive and permeable mutant Z114 was 59.1% and over 61% of the total protein could be released from the cells treated at 37 degrees C. The mutant cells grown in the yacon extract still contained high level of essential amino acids and other nutrients. This means that the yacon extract could be used as the medium for growth of the highly thermosensitive and permeable mutant which contained high content of crude protein.

  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.

  20. Membrane Vesicles Released by a hypervesiculating Escherichia coli Nissle 1917 tolR Mutant Are Highly Heterogeneous and Show Reduced Capacity for Epithelial Cell Interaction and Entry

    PubMed Central

    Pérez-Cruz, Carla; Cañas, María-Alexandra; Giménez, Rosa; Badia, Josefa; Mercade, Elena; Aguilera, Laura

    2016-01-01

    Membrane vesicles (MVs) produced by Gram-negative bacteria are being explored for novel clinical applications due to their ability to deliver active molecules to distant host cells, where they can exert immunomodulatory properties. MVs released by the probiotic Escherichia coli Nissle 1917 (EcN) are good candidates for testing such applications. However, a drawback for such studies is the low level of MV isolation from in vitro culture supernatants, which may be overcome by the use of mutants in cell envelope proteins that yield a hypervesiculation phenotype. Here, we confirm that a tolR mutation in EcN increases MV production, as determined by protein, LPS and fluorescent lipid measurements. Transmission electron microscopy (TEM) of negatively stained MVs did not reveal significant differences with wild type EcN MVs. Conversely, TEM observation after high-pressure freezing followed by freeze substitution of bacterial samples, together with cryo-TEM observation of plunge-frozen hydrated isolated MVs showed considerable structural heterogeneity in the EcN tolR samples. In addition to common one-bilayer vesicles (OMVs) and the recently described double-bilayer vesicles (O-IMVs), other types of MVs were observed. Time-course experiments of MV uptake in Caco-2 cells using rhodamine- and DiO-labelled MVs evidenced that EcN tolR MVs displayed reduced internalization levels compared to the wild-type MVs. The low number of intracellular MVs was due to a lower cell binding capacity of the tolR-derived MVs, rather than a different entry pathway or mechanism. These findings indicate that heterogeneity of MVs from tolR mutants may have a major impact on vesicle functionality, and point to the need for conducting a detailed structural analysis when MVs from hypervesiculating mutants are to be used for biotechnological applications. PMID:28036403

  1. 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.

  2. Efficient and high fidelity incorporation of dye-terminators by a novel archaeal DNA polymerase mutant.

    PubMed

    Arezi, Bahram; Hansen, Connie J; Hogrefe, Holly H

    2002-09-27

    We examined the molecular basis of ddNTP selectivity in archaeal family B DNA polymerases by randomly mutagenizing the gene encoding Thermococcus sp. JDF-3 DNA polymerase and screening mutant libraries for improved ddNTP incorporation. We identified two mutations, P410L and A485T, that improved ddNTP uptake, suggesting the contribution of P410 and A485 to ddNTP/dNTP selectivity in archaeal DNA polymerases. The importance of A485 was identified previously in mutagenesis studies employing Pfu (A486) and Vent (A488) DNA polymerases, while the contribution of P410 to ddNTP/dNTP selectivity has not been reported. We demonstrate that a combination of mutations (P410L/A485T) has an additive effect in improving ddNTP incorporation by a total of 250-fold. To assess the usefulness of the JDF-3 P410L/A485T in fluorescent-sequencing applications, we compared the archaeal mutant to Taq F667Y with respect to fidelity and kinetic parameters for DNA and dye-ddNTPs. Although the Taq F667Y and JDF-3 P410L/A485T mutants exhibit similar K(m) and V(max) values for dye-ddNTPs in single-base extension assays, the archaeal mutant exhibits higher fidelity due to a reduced tendency to form certain (ddG:dT, ddT:dC) mispairs. DNA polymerases exhibiting higher insertion fidelity are expected to provide greater accuracy in SNP frequency determinations by single-base extension and in multiplex minisequencing assays.

  3. 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.

  4. Mutants of Micromonospora viridifaciens sialidase have highly variable activities on natural and non-natural substrates.

    PubMed

    Jers, C; Guo, Y; Kepp, K P; Mikkelsen, J D

    2015-02-01

    This study aimed to improve the hydrolase activity of the well-characterised bacterial sialidase from Micromonospora viridifaciens. The enzyme and its mutated versions were produced in Bacillus subtilis and secreted to the growth medium. Twenty amino acid positions in or near the active site were subjected to site-saturation mutagenesis and evaluated on the artificial sialidase substrate 2-O-(p-nitrophenyl)-α-d-N-acetylneuraminic acid and on the natural substrate casein glycomacropeptide. A considerably higher fraction of the mutants exhibited increased activity on the artificial substrate compared with the natural one, with the most proficient mutant showing a 13-fold improvement in kcat/Km. In contrast, no mutants displayed more than a 2-fold increase in activity on the natural substrate. To gain further insight into this important discrepancy, we analysed the stability of mutants using the PoPMuSiC software, a property that also correlates with the potential for introducing chemical variation, after validating the method with a set of experimental stability estimates. We found a significant correlation between improved hydrolase activity on the artificial substrate and reduced apparent stability. Together with the minor improvement on the natural substrate this shows an important difference between naturally evolved functionality and new laboratory functionality. Our results suggest that when engineering sialidases and potentially other proteins towards non-natural substrates that are not optimized by natural evolution, major changes in chemical properties are advantageous, and these changes tend to correlate with decreased stability, partly explaining commonly observed trade-offs between stability and proficiency.

  5. A Mutant of Bacillus Subtilis with High-Producing Surfactin by Ion Beam Implantation

    NASA Astrophysics Data System (ADS)

    Liu, Qingmei; Yuan, Hang; Wang, Jun; Gong, Guohong; Zhou, Wei; Fan, Yonghong; Wang, Li; Yao, Jianming; Yu, Zengliang

    2006-07-01

    In order to generate a mutant of Bacillus subtilis with enhanced surface activity through low energy nitrogen ion beam implantation, the effects of energy and dose of ions implanted were studied. The morphological changes in the bacteria were observed by scanning electron microscope (SEM). The optimum condition of ions implantation, 20 keV of energy and 2.6×1015N+/cm2 in dose, was determined. A mutant, B.s-E-8 was obtained, whose surface activity of 50-fold and 100-fold diluted cell-free Landy medium was as 5.6-fold and 17.4-fold as the wild strain. The microbial growth and biosurfactant production of both the mutant and the wild strain were compared. After purified by ultrafiltration and SOURCE 15PHE, the biosurfactant was determined to be a complex of surfactin family through analysis of electrospray ionization mass spectrum (ESI/MS) and there was an interesting finding that after the ion beam implantation the intensities of the components were different from the wild type strain.

  6. 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.

  7. 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.

  8. Field-based high throughput phenotyping rapidly identifies genomic regions controlling yield components in rice

    PubMed Central

    Tanger, Paul; Klassen, Stephen; Mojica, Julius P.; Lovell, John T.; Moyers, Brook T.; Baraoidan, Marietta; Naredo, Maria Elizabeth B.; McNally, Kenneth L.; Poland, Jesse; Bush, Daniel R.; Leung, Hei; Leach, Jan E.; McKay, John K.

    2017-01-01

    To ensure food security in the face of population growth, decreasing water and land for agriculture, and increasing climate variability, crop yields must increase faster than the current rates. Increased yields will require implementing novel approaches in genetic discovery and breeding. Here we demonstrate the potential of field-based high throughput phenotyping (HTP) on a large recombinant population of rice to identify genetic variation underlying important traits. We find that detecting quantitative trait loci (QTL) with HTP phenotyping is as accurate and effective as traditional labor-intensive measures of flowering time, height, biomass, grain yield, and harvest index. Genetic mapping in this population, derived from a cross of an modern cultivar (IR64) with a landrace (Aswina), identified four alleles with negative effect on grain yield that are fixed in IR64, demonstrating the potential for HTP of large populations as a strategy for the second green revolution. PMID:28220807

  9. High-yield boron nitride nanosheets from 'chemical blowing': towards practical applications in polymer composites.

    PubMed

    Wang, Xuebin; Pakdel, Amir; Zhi, Chunyi; Watanabe, Kentaro; Sekiguchi, Takashi; Golberg, Dmitri; Bando, Yoshio

    2012-08-08

    An improved 'chemical blowing' route presuming atmospheric-pressure pre-treatment and moderate heating rate of designated precursors was developed to synthesize ultra-thin boron nitride (BN) nanosheets with high yield and large lateral dimensions. The yield reached as high as 40 wt% with respect to raw materials (ammonia borane). The strong oxygen-related ultraviolet luminescence together with a blue emission of these BN nanosheets was then documented and analyzed. This implies potential applications in solid-state lighting, ultraviolet lasing and full-color luminescence. Mechanical strength of different polymeric composites with a small fraction of BN nanosheet fillers was dramatically increased by tens of per cent, while high transparency of composite materials was still maintained in the visible optical range. The increased yield and reduced cost of BN nanosheets should promote their wide practical applications in various composites.

  10. 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.

  11. 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

  12. 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

  13. 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.

  14. Development of a Highly Sensitive Bioluminescent Enzyme Immunoassay for Hepatitis B Virus Surface Antigen Capable of Detecting Divergent Mutants

    PubMed Central

    Takehara, Shizuka; Takahashi, Masaharu

    2013-01-01

    Hepatitis B virus (HBV) infections are sometimes overlooked when using commercial kits to measure hepatitis B virus surface antigen (HBsAg) due to their low sensitivities and reactivities to mutant strains of various genotypes. We developed an ultrasensitive bioluminescent enzyme immunoassay (BLEIA) for HBsAg using firefly luciferase, which is adaptable to a variety of HBsAg mutants, by combining four monoclonal antibodies with a polyclonal antibody against HBsAg. The measurement of seroconversion panels showed trace amounts of HBsAg during the early infection phase by the BLEIA because of its high sensitivity of 5 mIU/ml. The BLEIA detected HBsAg as early as did PCR in five of seven series and from 2.1 to 9.4 days earlier than commercial immunoassay methods. During the late infection phase, the BLEIA successfully detected HBsAg even 40 days after the disappearance of HBV DNA and the emergence of antibodies against HBsAg. The HBsAg BLEIA successfully detected all 13 recombinant HBsAg and 45 types of HBsAg mutants with various mutations within amino acids 90 to 164 in the S gene product. Some specimens had higher values determined by the BLEIA than those by a commercial chemiluminescent immunoassay; this suggests that such discrepancies were caused by the dissociation of preS1/preS2 peptides from the particle surface. With its highly sensitive detection of low-titer HBsAg, including various mutants, the HBsAg BLEIA is considered to be useful for the early diagnosis and prevention of HBV infection because of the shorter window of infection prior to detection, which facilitates early prediction of recurrence in HBV-infected individuals. PMID:23761660

  15. 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.

  16. 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%.

  17. 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.

  18. 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.

  19. 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

  20. 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...

  1. 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...

  2. 31 CFR 356.21 - How are awards at the high yield or discount rate calculated?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 31 Money and Finance: Treasury 2 2010-07-01 2010-07-01 false How are awards at the high yield or discount rate calculated? 356.21 Section 356.21 Money and Finance: Treasury Regulations Relating to Money and Finance (Continued) FISCAL SERVICE, DEPARTMENT OF THE TREASURY BUREAU OF THE PUBLIC DEBT SALE...

  3. Dakota Diamond: An exceptionally high yielding, cold chipping potato cultivar with long-term storage potential

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Dakota Diamond (ND5822C-7) is a medium to late maturing cultivar with uniformly sized tubers and very high yield potential. It resulted from the cross of ND4103-2 and “Dakota Pearl”. Dakota Diamond is comprised of approximately 23.3% wild potato species germplasm. It combines the characteristics ...

  4. Surrogate models for identifying robust, high yield regions of parameter space for ICF implosion simulations

    NASA Astrophysics Data System (ADS)

    Humbird, Kelli; Peterson, J. Luc; Brandon, Scott; Field, John; Nora, Ryan; Spears, Brian

    2016-10-01

    Next-generation supercomputer architecture and in-transit data analysis have been used to create a large collection of 2-D ICF capsule implosion simulations. The database includes metrics for approximately 60,000 implosions, with x-ray images and detailed physics parameters available for over 20,000 simulations. To map and explore this large database, surrogate models for numerous quantities of interest are built using supervised machine learning algorithms. Response surfaces constructed using the predictive capabilities of the surrogates allow for continuous exploration of parameter space without requiring additional simulations. High performing regions of the input space are identified to guide the design of future experiments. In particular, a model for the yield built using a random forest regression algorithm has a cross validation score of 94.3% and is consistently conservative for high yield predictions. The model is used to search for robust volumes of parameter space where high yields are expected, even given variations in other input parameters. Surrogates for additional quantities of interest relevant to ignition are used to further characterize the high yield regions. This work performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344, Lawrence Livermore National Security, LLC. LLNL-ABS-697277.

  5. Optimum doping achieves high quantum yields in GaAs photoemitters

    NASA Technical Reports Server (NTRS)

    Sonnenberg, H.

    1971-01-01

    Experimental data indicate that optimum doping exists. Measured quantum yield curves indicate optimum overall response is obtained in GaAs emitters with doping in high 10 to the 18th power per cu cm range. Doping for optimum response is not necessarily in this range.

  6. 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

  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. Extracting DNA from 'jaws': high yield and quality from archived tiger shark (Galeocerdo cuvier) skeletal material.

    PubMed

    Nielsen, E E; Morgan, J A T; Maher, S L; Edson, J; Gauthier, M; Pepperell, J; Holmes, B J; Bennett, M B; Ovenden, J R

    2016-08-10

    Archived specimens are highly valuable sources of DNA for retrospective genetic/genomic analysis. However, often limited effort has been made to evaluate and optimize extraction methods, which may be crucial for downstream applications. Here, we assessed and optimized the usefulness of abundant archived skeletal material from sharks as a source of DNA for temporal genomic studies. Six different methods for DNA extraction, encompassing two different commercial kits and three different protocols, were applied to material, so-called bio-swarf, from contemporary and archived jaws and vertebrae of tiger sharks (Galeocerdo cuvier). Protocols were compared for DNA yield and quality using a qPCR approach. For jaw swarf, all methods provided relatively high DNA yield and quality, while large differences in yield between protocols were observed for vertebrae. Similar results were obtained from samples of white shark (Carcharodon carcharias). Application of the optimized methods to 38 museum and private angler trophy specimens dating back to 1912 yielded sufficient DNA for downstream genomic analysis for 68% of the samples. No clear relationships between age of samples, DNA quality and quantity were observed, likely reflecting different preparation and storage methods for the trophies. Trial sequencing of DNA capture genomic libraries using 20 000 baits revealed that a significant proportion of captured sequences were derived from tiger sharks. This study demonstrates that archived shark jaws and vertebrae are potential high-yield sources of DNA for genomic-scale analysis. It also highlights that even for similar tissue types, a careful evaluation of extraction protocols can vastly improve DNA yield.

  9. High-yield electrochemical production of formaldehyde from CO2 and seawater.

    PubMed

    Nakata, Kazuya; Ozaki, Takuya; Terashima, Chiaki; Fujishima, Akira; Einaga, Yasuaki

    2014-01-13

    The catalytic, electrocatalytic, or photocatalytic conversion of CO2 into useful chemicals in high yield for industrial applications has so far proven difficult. Herein, we present our work on the electrochemical reduction of CO2 in seawater using a boron-doped diamond (BDD) electrode under ambient conditions to produce formaldehyde. This method overcomes the usual limitation of the low yield of higher-order products, and also reduces the generation of H2 . In comparison with other electrode materials, BDD electrodes have a wide potential window and high electrochemical stability, and, moreover, exhibit very high Faradaic efficiency (74%) for the production of formaldehyde, using either methanol, aqueous NaCl, or seawater as the electrolyte. The high Faradaic efficiency is attributed to the sp(3)-bonded carbon of the BDD. Our results have wide ranging implications for the efficient and cost-effective conversion of CO2.

  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 Central

    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. 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.

  14. High precision and high yield fabrication of dense nanoparticle arrays onto DNA origami at statistically independent binding sites

    NASA Astrophysics Data System (ADS)

    Takabayashi, Sadao; Klein, William P.; Onodera, Craig; Rapp, Blake; Flores-Estrada, Juan; Lindau, Elias; Snowball, Lejmarc; Sam, Joseph T.; Padilla, Jennifer E.; Lee, Jeunghoon; Knowlton, William B.; Graugnard, Elton; Yurke, Bernard; Kuang, Wan; Hughes, William L.

    2014-10-01

    High precision, high yield, and high density self-assembly of nanoparticles into arrays is essential for nanophotonics. Spatial deviations as small as a few nanometers can alter the properties of near-field coupled optical nanostructures. Several studies have reported assemblies of few nanoparticle structures with controlled spacing using DNA nanostructures with variable yield. Here, we report multi-tether design strategies and attachment yields for homo- and hetero-nanoparticle arrays templated by DNA origami nanotubes. Nanoparticle attachment yield via DNA hybridization is comparable with streptavidin-biotin binding. Independent of the number of binding sites, >97% site-occupation was achieved with four tethers and 99.2% site-occupation is theoretically possible with five tethers. The interparticle distance was within 2 nm of all design specifications and the nanoparticle spatial deviations decreased with interparticle spacing. Modified geometric, binomial, and trinomial distributions indicate that site-bridging, steric hindrance, and electrostatic repulsion were not dominant barriers to self-assembly and both tethers and binding sites were statistically independent at high particle densities.High precision, high yield, and high density self-assembly of nanoparticles into arrays is essential for nanophotonics. Spatial deviations as small as a few nanometers can alter the properties of near-field coupled optical nanostructures. Several studies have reported assemblies of few nanoparticle structures with controlled spacing using DNA nanostructures with variable yield. Here, we report multi-tether design strategies and attachment yields for homo- and hetero-nanoparticle arrays templated by DNA origami nanotubes. Nanoparticle attachment yield via DNA hybridization is comparable with streptavidin-biotin binding. Independent of the number of binding sites, >97% site-occupation was achieved with four tethers and 99.2% site-occupation is theoretically possible with five

  15. High-yield production of functional soluble single-domain antibodies in the cytoplasm of Escherichia coli

    PubMed Central

    2013-01-01

    Background For their application in the area of diagnosis and therapy, single-domain antibodies (sdAbs) offer multiple advantages over conventional antibodies and fragments thereof in terms of size, stability, solubility, immunogenicity, production costs as well as tumor uptake and blood clearance. Thus, sdAbs have been identified as valuable next-generation targeting moieties for molecular imaging and drug delivery in the past years. Since these probes are much less complex than conventional antibody fragments, bacterial expression represents a facile method in order to produce sdAbs in large amounts as soluble and functional proteins. Results By the combined use of high cell density cultivation media with a genetically engineered E. coli mutant strain designed for the cytoplasmic formation of proper disulfide bonds, we achieved high level of intracellular sdAb production (up to 200 mg/L). Due to a carboxyterminal hexahistidine epitope, the soluble recombinant sdAbs could be purified by one-step immobilized metal affinity chromatography to apparent homogeneity and easily radiolabeled with 99mTc within 1 h. The intradomain disulfide bridge being critical for the stability and functionality of the sdAb molecule was shown to be properly formed in ~96% of the purified proteins. In vitro binding studies confirmed the high affinity and specificity of the expressed sdAb 7C12 towards its molecular target. Conclusions Our study demonstrates an efficient cultivation and expression strategy for the production of substantial amounts of soluble and functional sdAbs, which may be adopted for high-yield production of other more complex proteins with multiple disulfides as well. PMID:24161153

  16. Elevated Levels of High-Melting-Point Phosphatidylglycerols Do Not Induce Chilling Sensitivity in an Arabidopsis Mutant.

    PubMed Central

    Wu, J.; Browse, J.

    1995-01-01

    Molecular species of phosphatidylglycerol that contain only 16:0, 18:0, and 16:1-trans fatty acids undergo the transition from liquid crystalline phase to gel phase at temperatures well above 20[deg]C. Several lines of evidence have been used to implicate elevated proportions of these high-melting-point molecular species as a major cause of plant chilling sensitivity. In the fatty acid biosynthesis 1 (fab1) mutant of Arabidopsis, leaf phosphatidylglycerol contained 43% high-melting-point molecular species[mdash]a higher percentage than is found in many chilling-sensitive plants. Nevertheless, the mutant was completely unaffected (when compared with wild-type controls) by a range of low-temperature treatments that quickly led to the death of cucumber and other chilling-sensitive plants. Our results clearly demonstrate that high-melting-point phosphatidylglycerols do not mediate classic chilling damage. However, growth of fab1 plants was compromised by long-term (>2 weeks) exposure to 2[deg]C. This finding and other observations are consistent with a proposition that plants native to tropical and subtropical regions have evolved many traits that are incompatible with long-term growth or development in cooler climates but that may confer selective advantages at high temperatures. PMID:12242349

  17. 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

  18. 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.

  19. 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

  20. 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.

  1. 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.

  2. 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.

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

    PubMed Central

    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

    2014-01-01

    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. PMID:24151938

  4. 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.

  5. 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 ...

  6. 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.

  7. High Precision and High Yield Fabrication of Dense Nanoparticle Arrays onto DNA Origami at Statistically Independent Binding Sites †

    PubMed Central

    Takabayashi, Sadao; Klein, William P.; Onodera, Craig; Rapp, Blake; Flores-Estrada, Juan; Lindau, Elias; Snowball, Lejmarc; Sam, Joseph Tyler; Padilla, Jennifer E.; Lee, Jeunghoon; Knowlton, William B.; Graugnard, Elton; Yurke, Bernard; Kuang, Wan; Hughes, William L.

    2015-01-01

    High precision, high yield, and high density self-assembly of nanoparticles into arrays is essential for nanophotonics. Spatial deviations as small as a few nanometers can alter the properties of near-field coupled optical nanostructures. Several studies have reported assemblies of few nanoparticle structures with controlled spacing using DNA nanostructures with variable yield. Here, we report multi-tether design strategies and attachment yields for homo- and hetero-nanoparticle arrays templated by DNA origami nanotubes. Nanoparticle attachment yield via DNA hybridization is comparable with streptavidin-biotin binding. Independent of the number of binding sites, >97% site-occupation was achieved with four tethers and 99.2% site-occupation is theoretically possible with five tethers. The interparticle distance was within 2 nm of all design specifications and the nanoparticle spatial deviations decreased with interparticle spacing. Modified geometric, binomial, and trinomial distributions indicate that site-bridging, steric hindrance, and electrostatic repulsion were not dominant barriers to self-assembly and both tethers and binding sites were statistically independent at high particle densities. PMID:25311051

  8. [Molecular ecological basis of high-yielding formation of rice and its application].

    PubMed

    Lin, Wenxiong; Liang, Kangjing; Guo, Yuchun; He, Huaqin; Wang, Jingyuan; Liang, Yiyuan; Chen, Fangyu

    2003-12-01

    This paper introduced the developmental genetics and its molecular ecological basis of high yielding formation of rice in the past decade, and analyzed the advantage and the shortage of comparative physiological approach traditionally used in the research work on crop cultivation. It was emphasized to actively introduce the research contents and its methodology from relative disciplines to deeply understand the scientific issue, and suggested that the key to realize stable and high yielding of rice was to develop a rational cultivation system based on the properties of genetic effects on the traits in different developmental stages by controlling and regulating the traits governed by dominant effect genes and additive effect genes x environment in same direction, which was considered as the main characteristics and the technological innovation of modern crop genetic ecological cultivation science. Finally, the development trend of crop cultivation science shifting to molecular crop cultivation science was predicted and discussed.

  9. Detection of two major grain yield QTL in bread wheat (Triticum aestivum L.) under heat, drought and high yield potential environments.

    PubMed

    Bennett, Dion; Reynolds, Matthew; Mullan, Daniel; Izanloo, Ali; Kuchel, Haydn; Langridge, Peter; Schnurbusch, Thorsten

    2012-11-01

    A large proportion of the worlds' wheat growing regions suffers water and/or heat stress at some stage during the crop growth cycle. With few exceptions, there has been no utilisation of managed environments to screen mapping populations under repeatable abiotic stress conditions, such as the facilities developed by the International Wheat and Maize Improvement Centre (CIMMYT). Through careful management of irrigation and sowing date over three consecutive seasons, repeatable heat, drought and high yield potential conditions were imposed on the RAC875/Kukri doubled haploid population to identify genetic loci for grain yield, yield components and key morpho-physiological traits under these conditions. Two of the detected quantitative trait loci (QTL) were located on chromosome 3B and had a large effect on canopy temperature and grain yield, accounting for up to 22 % of the variance for these traits. The locus on chromosome arm 3BL was detected under all three treatments but had its largest effect under the heat stress conditions, with the RAC875 allele increasing grain yield by 131 kg ha(-1) (or phenotypically, 7 % of treatment average). Only two of the eight yield QTL detected in the current study (including linkage groups 3A, 3D, 4D 5B and 7A) were previously detected in the RAC875/Kukri doubled haploid population; and there were also different yield components driving grain yield. A number of discussion points are raised to understand differences between the Mexican and southern Australian production environments and explain the lack of correlation between the datasets. The two key QTL detected on chromosome 3B in the present study are candidates for further genetic dissection and development of molecular markers.

  10. 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

  11. Extraction conditions of Antheraea mylitta sericin with high yields and minimum molecular weight degradation.

    PubMed

    Yun, Haesung; Oh, Hanjin; Kim, Moo Kon; Kwak, Hyo Won; Lee, Jeong Yun; Um, In Chul; Vootla, Shyam Kumar; Lee, Ki Hoon

    2013-01-01

    Although the technique for extracting the Bombyx mori sericin has been extensively known, the extraction of sericin from wild-silkworm cocoons is not yet standardized. The aim of this study was to find the optimal conditions for the extraction of sericin from Antheraea mylitta cocoons, with high yields and minimum degradation. We attempted to apply various protocols for the extraction of the A. mylitta sericin (AmS). Among these, we found that the extraction of AmS with a sodium carbonate solution exhibited the highest yield except the conventional soap-alkali extraction. To find the optimal conditions for the AmS extraction with the sodium carbonate, we changed the concentration of sodium carbonate and the treatment time. With an increase in the sodium carbonate concentration and the extraction time, the yield of AmS increased, but the molecular weight (MW) of AmS decreased. Considering the yield, molecular weight distribution (MWD) and amino acid composition of AmS, we suggest that the optimal conditions for the AmS extraction require treatment with 0.02 M sodium carbonate and boiling for 60 min.

  12. 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

  13. 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.

  14. Competitive growth experiments with a high-lipid Chlamydomonas reinhardtii mutant strain and its wild-type to predict industrial and ecological risks.

    PubMed

    Russo, David A; Beckerman, Andrew P; Pandhal, Jagroop

    2017-12-01

    Key microalgal species are currently being exploited as biomanufacturing platforms using mass cultivation systems. The opportunities to enhance productivity levels or produce non-native compounds are increasing as genetic manipulation and metabolic engineering tools are rapidly advancing. Regardless of the end product, there are both environmental and industrial risks associated to open pond cultivation of mutant microalgal strains. A mutant escape could be detrimental to local biodiversity and increase the risk of algal blooms. Similarly, if the cultivation pond is invaded by a wild-type (WT) microalgae or the mutant reverts to WT phenotypes, productivity could be impacted. To investigate these potential risks, a response surface methodology was applied to determine the competitive outcome of two Chlamydomonas reinhardtii strains, a WT (CC-124) and a high-lipid accumulating mutant (CC-4333), grown in mixotrophic conditions, with differing levels of nitrogen and initial WT to mutant ratios. Results of the growth experiments show that mutant cells have double the exponential growth rate of the WT in monoculture. However, due to a slower transition from lag phase to exponential phase, mutant cells are outcompeted by the WT in every co-culture treatment. This suggests that, under the conditions tested, outdoor cultivation of the C. reinhardtii cell wall-deficient mutant strains does not carry a significant environmental risk to its WT in an escape scenario. Furthermore, lipid results show the mutant strain accumulates over 200% more TAGs per cell, at 50 mg L(-1) NH4Cl, compared to the WT, therefore, the fragility of the mutant strain could impact on overall industrial productivity.

  15. 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.

  16. 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

  17. 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.

  18. 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.

  19. 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.

  20. Comparative Transcriptome Analysis Using High Papaverine Mutant of Papaver somniferum Reveals Pathway and Uncharacterized Steps of Papaverine Biosynthesis

    PubMed Central

    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. PMID:23738019

  1. 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

  2. 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.

  3. Behavioural effects of high fat diet in a mutant mouse model for the schizophrenia risk gene neuregulin 1.

    PubMed

    Holm-Hansen, S; Low, J K; Zieba, J; Gjedde, A; Bergersen, L H; Karl, T

    2016-03-01

    Schizophrenia patients are often obese or overweight and poor dietary choices appear to be a factor in this phenomenon. Poor diet has been found to have complex consequences for the mental state of patients. Thus, this study investigated whether an unhealthy diet [i.e. high fat diet (HFD)] impacts on the behaviour of a genetic mouse model for the schizophrenia risk gene neuregulin 1 (i.e. transmembrane domain Nrg1 mutant mice: Nrg1 HET). Female Nrg1 HET and wild-type-like littermates (WT) were fed with either HFD or a control chow diet. The mice were tested for baseline (e.g. anxiety) and schizophrenia-relevant behaviours after 7 weeks of diet exposure. HFD increased body weight and impaired glucose tolerance in all mice. Only Nrg1 females on HFD displayed a hyper-locomotive phenotype as locomotion-suppressive effects of HFD were only evident in WT mice. HFD also induced an anxiety-like response and increased freezing in the context and the cued version of the fear conditioning task. Importantly, CHOW-fed Nrg1 females displayed impaired social recognition memory, which was absent in HFD-fed mutants. Sensorimotor gating deficits of Nrg1 females were not affected by diet. In summary, HFD had complex effects on the behavioural phenotype of test mice and attenuated particular cognitive deficits of Nrg1 mutant females. This topic requires further investigations thereby also considering other dietary factors of relevance for schizophrenia as well as interactive effects of diet with medication and sex.

  4. High yields and soluble expression of superoxide dismutases in Escherichia coli due to the HIV-1 Tat peptide via increases in mRNA transcription

    PubMed Central

    Sun, Yangdong; Ye, Qiao; Wu, Min; Wu, Yonghong; Zhang, Chenggang; Yan, Weiqun

    2016-01-01

    This study aimed to validate the high yield and soluble expression of proteins carrying the transactivator of transcription (Tat) peptide tag, and further explored the potential mechanism by which the Tat tag increases expression. Escherichia coli superoxide dismutase (SOD) proteins, including SodA, SodB and SodC, were selected for analysis. As expected, the yields and the solubility of Tat-tagged proteins were higher than those of Tat-free proteins, and similar results were observed for the total SOD enzyme activity. Bacterial cells that overexpressed Tat-tagged proteins exhibited increased anti-paraquat activity compared with those expressing Tat-free proteins that manifested as SodA>SodC>SodB. When compared with an MG1655 wild-type strain, the growth of a ΔSodA mutant strain was found to be inhibited after paraquat treatment; the growth of ΔSodB and ΔSodC mutant strains was also slightly inhibited. The mRNA transcript level of genes encoding Tat-tagged proteins was higher than that of genes encoding Tat-free proteins. Furthermore, the α-helix and turn of Tat-tagged proteins were higher than those of Tat-free proteins, but the β-sheet and random coil content was lower. These results indicated that the incorporation of the Tat core peptide as a significant basic membrane transduction peptide in fusion proteins could increase mRNA transcripts and promote the high yield and soluble expression of heterologous proteins in E. coli. PMID:27741225

  5. 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

  6. 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.

  7. High frequency of mosaic mutants produced by N-ethyl-N-nitrosourea exposure of mouse zygotes.

    PubMed

    Russell, L B; Bangham, J W; Stelzner, K F; Hunsicker, P R

    1988-12-01

    Mouse zygotes containing one multiple-recessive parental genome (a, b; p cch; d se; s) and the corresponding wild-type alleles in the other were exposed to N-ethyl-N-nitrosourea (ENU) at various stages in vivo. At weaning age, the resulting mice were examined for mutations at the marked loci as well as at others producing externally visible phenotypes. Because of viability problems in one of two reciprocal crosses, the bulk of the mutagenesis data are derived from the cross that detects recessive mutations in the maternal genome. The mutation rate was approximately 8 times higher in groups treated 2.5-3 hr postmating (sperm entry, completion of second meiotic division) than in those injected 5-6 hr postmating (pronuclear formation). In the former more sensitive zygote population, the mutation rate is about an order of magnitude greater than that induced by the same ENU exposure (50 mg/kg) to spermatogonial stem cells. Of 11 mutants recovered, 8 were mosaics. Progeny tests have demonstrated germ-line involvement for most of the mosaics, and the average fraction of the germ line carrying the mutation is close to 50%. The nature of the mutations indicates (i) that the mosaicism results not from misassortment at the first cleavage but from mutation affecting one DNA strand of the maternal chromosome, and (ii) that the mutations are intragenic lesions rather than multilocus deletions, thus resembling ENU-induced mutations in spermatogonia. The finding that mosaicism for presumed point mutations is readily inducible by ENU treatment of zygotes may provide a means of generating genetic materials that can be of use for developmental studies.

  8. High Titer and Yields Achieved with Novel, Low-Severity Pretreatment Strategy

    SciTech Connect

    2016-03-01

    NREL researchers obtained high concentration sugar syrups in enzymatic hydrolysis that are fermentable to ethanol and other advanced biofuels and intermediate products at high yields. The novel DMR process is simpler and bypasses all severe pretreatment methods, thus reducing the environmental impact. The results are unprecedented. Researchers achieved a high concentration of sugars (230g/L of monomeric sugar and 270 g/L total sugar) and this low toxicity, highly fermentable syrup yielded 86 g/L ethanol (> 90 percent conversion). In addition, the lignin streams from this process can readily be converted to jet or renewable diesel blendstocks through a hydrodeoxygenation step. The NREL-developed, low severity DMR process may potentially replace higher severity chemical pretreatments and associated expensive reactors constructed of exotic alloys with a simpler process, using commercial-scale equipment commonly associated with the pulp and paper industry, to produce high concentration, low toxicity sugar streams and highly reactive lignin streams from non-food renewable biomass for biological and catalytic upgrading to advanced biofuels and chemicals. The simpler DMR process with black liquor recycling could reduce environmental and life-cycle impacts, and repurpose shuttered pulp and paper mills to help revitalize rural economies.

  9. Preparation of ammonia borane in high yield and purity, methanolysis, and regeneration.

    PubMed

    Ramachandran, P Veeraraghavan; Gagare, Pravin D

    2007-09-17

    Ammonia borane (AB) is emerging as a promising solid hydrogen carrier, particularly for power generation in portable devices that employ proton-exchange membrane fuel cells. A preparative-scale synthesis of AB from sodium borohydride and ammonium salts in high yields (> or =95%) and very high purity (> or =98%) has been described. The first systematic study of a transitional metal-catalyzed alcoholysis of AB, comparison of the methanolysis to the hydrolysis of AB, and regeneration of AB from ammonium tetramethoxyborate also has been described.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. Improvements in immunoprecipitation of specific messenger RNA. Isolation of highly purified conalbumin mRNA in high yield.

    PubMed

    Payvar, F; Schimke, R T

    1979-11-01

    We have described previously procedures for the isolation of specific mRNA employing immunoprecipitation of polysomes. In spite of our success with ovalbumin mRNA in the chicken oviduct, we have had considerable difficulties in applying these same published techniques to the immunopurification of conalbumin mRNA, despite the fact that the chicken oviduct synthesizes up to 10% of protein as conalbumin. Here we describe a number of modifications and refinements which have proved essential in obtaining intact conalbumin mRNA in high purity and high yields. These refinements include: (a) improved purification of conalbumin in order to remove contaminating proteins that result in impure antibodies; (b) improved isolation of specific conalbumin antibody in high yields; (c) improved methods for reducing contamination by non-specific polysomes; (d) improved techniques for isolation of RNA from immunoprecipitates resulting in less degradation and higher recovery of conalbumin mRNA; (E) improved techniques for efficient translation of conalbumin mRNA involving treatment of the RNA with methylmercury prior to translation. We conclude that problems involved in the immunoprecipitation of different mRNAs may differ, and that various refinements in techniques may be required for obtaining highly purified preparations of intact mRNA in high yields.

  15. Streptococcus pyogenes Sortase Mutants Are Highly Susceptible to Killing by Host Factors Due to Aberrant Envelope Physiology

    PubMed Central

    Raz, Assaf; Tanasescu, Ana-Maria; Zhao, Anna M.; Serrano, Anna; Alston, Tricia; Sol, Asaf; Bachrach, Gilad; Fischetti, Vincent A.

    2015-01-01

    Cell wall anchored virulence factors are critical for infection and colonization of the host by Gram-positive bacteria. Such proteins have an N-terminal leader sequence and a C-terminal sorting signal, composed of an LPXTG motif, a hydrophobic stretch, and a few positively charged amino acids. The sorting signal halts translocation across the membrane, allowing sortase to cleave the LPXTG motif, leading to surface anchoring. Deletion of sortase prevents the anchoring of virulence factors to the wall; the effects on bacterial physiology however, have not been thoroughly characterized. Here we show that deletion of Streptococcus pyogenes sortase A leads to accumulation of sorting intermediates, particularly at the septum, altering cellular morphology and physiology, and compromising membrane integrity. Such cells are highly sensitive to cathelicidin, and are rapidly killed in blood and plasma. These phenomena are not a loss-of-function effect caused by the absence of anchored surface proteins, but specifically result from the accumulation of sorting intermediates. Reduction in the level of sorting intermediates leads to a return of the sortase mutant to normal morphology, while expression of M protein with an altered LPXTG motif in wild type cells leads to toxicity in the host environment, similar to that observed in the sortase mutant. These unanticipated effects suggest that inhibition of sortase by small-molecule inhibitors could similarly lead to the rapid elimination of pathogens from an infected host, making such inhibitors much better anti-bacterial agents than previously believed. PMID:26484774

  16. Optimization of Escherichia coli cultivation methods for high yield neuropeptide Y receptor type 2 production.

    PubMed

    Berger, Christian; Montag, Cindy; Berndt, Sandra; Huster, Daniel

    2011-03-01

    The recombinant expression of human G protein-coupled receptors usually yields low production levels using commonly available cultivation protocols. Here, we describe the development of a high yield production protocol for the human neuropeptide Y receptor type 2 (Y2R), which provides the determination of expression levels in a time, media composition, and process parameter dependent manner. Protein was produced by Escherichia coli in a defined medium composition suitable for isotopic labeling required for investigations by nuclear magnetic resonance spectroscopy. The Y2 receptor was fused to a C-terminal 8x histidine tag by means of the pET vector system for easy one-step purification via affinity chromatography, yielding a purity of 95-99% for every condition tested, which was determined by SDS-PAGE and Western blot analysis. The Y2 receptor was expressed as inclusion body aggregates in complex media and minimal media, using different carbon sources. We investigated the influences of media composition, temperature, pH, and set specific growth rate on cell behavior, biomass wet weight specific and culture volume specific amounts of the target protein, which had been identified by inclusion body preparation, solubilization, followed by purification and spectrometric determination of the protein concentration. The developed process control strategy led to very high reproducibility of cell growth and protein concentrations with a maximum yield of 800 μg purified Y2 receptor per gram wet biomass when glycerol was used as carbon source in the mineral salt medium composition (at 38 °C, pH 7.0, and a set specific growth rate of 0.14 g/(gh)). The maximum biomass specific amount of purified Y2 receptor enabled the production of 35 mg Y2R per liter culture medium at an optical density (600 nm) of 25.

  17. Water mobility in the endosperm of high beta-glucan barley mutants as studied by nuclear magnetic resonance imaging.

    PubMed

    Fast Seefeldt, Helene; van den Berg, Frans; Köckenberger, Walter; Engelsen, Søren Balling; Wollenweber, Bernd

    2007-04-01

    (1)H NMR imaging (MRI) was used as a noninvasive technique to study water distribution and mobility in hydrated barley (Hordeum vulgare L.) seeds of accessions with varying content of beta glucan (BG), a highly hygroscopic cell wall component. High contents of BG in barley are unfavorable in malting where it leads to clotting of filters and hazing of beer as well as in animal feed where it hinders the rapid uptake of energy. However, a high content of BG has a positive nutritional effect, as it lowers the cholesterol and the glycaemic index. It was studied whether water distribution and mobility were related to content and location of BG. Water mobility was investigated by following the rate and mode of desiccation in hydrated single seeds. In order to determine the different water components, a multispin echo experiment was set up to reveal the T(2) transverse relaxation rates of water within the seeds. A principal component analysis (PCA) discriminated control seeds from the high-BG mutant seeds. MRI proved efficient in tracing the differences in water-holding capacity of contrasting barley seeds. All accessions showed nonuniform distribution of water at full hydration as well as during desiccation. The embryo retained water even after 36 h of drying, whereas the endosperm showed low and heterogeneous mobility of the water after drying. The relaxation time constants indicated that the BG mutants had regions of much higher water mobility around the ventral crease compared to the control. It is concluded that MRI can be applied to investigate temporal and spatial differences in the location of specific chemical compounds in single seeds.

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

    PubMed

    Taher, Edris; Chandran, Kartik

    2013-04-02

    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.

  19. 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

  20. 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

  1. 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.

  2. Classical effect for enhanced high harmonic yield in ultrashort laser pulses with a moderate laser intensity

    NASA Astrophysics Data System (ADS)

    Shi, Y. Z.; Wang, S.; Dong, F. L.; Li, Y. P.; Chen, Y. J.

    2017-03-01

    We study the influence of pulse duration on high harmonic generation (HHG) by exploring a wide laser-parameter region theoretically. Previous studies have shown that for high laser intensities close to saturation ionization intensity, the HHG inversion efficiency is higher for shorter pulses since the ground-state depletion is weaker in short pulses. Our simulations show that this high efficiency also appears for a moderate laser intensity at which the ionization is not very strong. A classical effect relating to shorter travel distances of the rescattering electron in shorter pulses is shown to contribute importantly to this high efficiency. The effect can be amplified significantly if a two-color laser field is used, suggesting a potential approach to increasing the HHG yield and generating short and bright attosecond pulses.

  3. 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.

  4. High-yield expression in Escherichia coli of soluble human MT2A with native functions.

    PubMed

    Yang, Fang; Zhou, Min; He, Zhimin; Liu, Xiaorong; Sun, Lin; Sun, Yu; Chen, Zhuchu

    2007-05-01

    Metallothioneins (MTs) are a family of low molecular weight, cysteine rich heavy metal binding proteins with multifunction, such as metal detoxification and antioxidation, and are involved in a number of cellular processes including gene expression, apoptosis, proliferation and differentiation. However, high yield expression of human MT in Escherichia coli has not been established effectively. To produce large amounts of human MT protein at low cost, recombinant human metallothionein 2A (MT2A) protein with an N-terminal GST tag was successfully expressed at high levels in soluble form in E. coli and high purification of it was established by affinity chromatography under native conditions. The final yield was about 5mg of the recombinant MT2A per liter of bacterial culture with the purity of 97.9%. Chemical and functional characteristics analysis of the recombinant human MT2A exhibited intact metal binding ability, hydroxyl radical scavenging ability and significant protective role against DNA damage caused by UVC radiation. Establishment of highly purified recombinant human MT2A protein with native characteristics at low cost would improve its function study and wide applications in protecting against oxidative damage and UV radiation.

  5. High incidence of lung, bone, and lymphoid tumors in transgenic mice overexpressing mutant alleles of the p53 oncogene.

    PubMed Central

    Lavigueur, A; Maltby, V; Mock, D; Rossant, J; Pawson, T; Bernstein, A

    1989-01-01

    We have investigated the role of the p53 gene in oncogenesis in vivo by generating transgenic mice carrying murine p53 genomic fragments isolated from a mouse Friend erythroleukemia cell line or BALB/c mouse liver DNA. Elevated levels of p53 mRNA were detected in several tissues of two transgenic lines tested. Increased levels of p53 protein were also detected in most of the tissues analyzed by Western blotting (immunoblotting). Because both transgenes encoded p53 proteins that were antigenically distinct from wild-type p53, it was possible to demonstrate that overexpression of the p53 protein was mostly, if not entirely, due to the expression of the transgenes. Neoplasms developed in 20% of the transgenic mice, with a high incidence of lung adenocarcinomas, osteosarcomas, and lymphomas. Tissues such as ovaries that expressed the transgene at high levels were not at higher risk of malignant transformation than tissues expressing p53 protein at much lower levels. The long latent period and low penetrance suggest that overexpression of p53 alone is not sufficient to induce malignancies and that additional events are required. These observations provide direct evidence that mutant alleles of the p53 oncogene have oncogenic potential in vivo and that different cell types show intrinsic differences in susceptibility to malignant transformation by p53. Since recent data suggest that p53 may be a recessive oncogene, it is possible that the elevated tumor incidence results from functional inactivation of endogenous p53 by overexpression of the mutant transgene. The high incidence of lung and bone tumors suggests that p53 transgenic mice may provide a useful model to investigate the molecular events that underlie these malignancies in humans. Images PMID:2476668

  6. Application of laboratory results to the design of a high yield VMIS oil shale retort

    SciTech Connect

    Bickel, T.C.; Ricketts, T.E.

    1986-01-01

    In situ oil shale retorts have typically been designed to process a rubble bed having uniform cross-sectional rubble properties. Edge effects during rock fragmentation commonly produce increased void at the perimeter of these low-void retorts. Previous laboratory and field results have demonstrated this void variation normal to the direction of flow causes non-uniform retort front velocities that result in significantly lower oil yield. It is unlikely that process control parameters (e.g., multiple injection points, steam, etc.) can provide any significant yield improvement in these non-uniform retorts. Any large improvement would come from modified rubblization concepts. This paper describes a modification to the retort blast design to achieve a uniform retorting front velocity in rubble with non-uniform properties (void fraction and particle size). This concept requires the creation of an anisotropic rubble bed with varying particle size and void fraction normal to the direction of flow. The unavoidable increased void at the retort perimeter is offset by modifying the ratio of the effective particle size of the rubble in the central to the perimeter regions of the retort. The results of laboratory-scale pressure drop and retorting experiments with an empirical blast design technique are used to describe how a high-yield, second generation in situ retort would be designed. 12 refs., 7 figs.

  7. 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.

  8. A high throughput DNA extraction method with high yield and quality

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Preparation of large quantity and high quality genomic DNA from a large number of plant samples is a major bottleneck for most genetic and genomic analyses, such as, genetic mapping, TILLING (Targeting Induced Local Lesion IN Genome), and next-generation sequencing directly from sheared ...

  9. 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

  10. 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.

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. 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.

  16. Pea DNA helicase 45 overexpression in tobacco confers high salinity tolerance without affecting yield.

    PubMed

    Sanan-Mishra, Neeti; Pham, Xuan Hoi; Sopory, Sudhir K; Tuteja, Narendra

    2005-01-11

    Salt tolerance is an important trait that is required to overcome salinity-induced reduction in plant productivity. We have reported previously the isolation of a pea DNA helicase 45 (PDH45) that exhibits striking homology with the eukaryotic translation initiation factor eIF-4A. Here, we report that PDH45 mRNA is induced in pea seedlings in response to high salt, and its overexpression driven by a constitutive cauliflower mosaic virus-(35)S promoter in tobacco plants confers salinity tolerance, thus suggesting a previously undescribed pathway for manipulating stress tolerance in crop plants. The T(0) transgenic plants showed high levels of PDH45 protein in normal and stress conditions, as compared with WT plants. The T(0) transgenics also showed tolerance to high salinity as tested by a leaf disk senescence assay. The T(1) transgenics were able to grow to maturity and set normal viable seeds under continuous salinity stress without any reduction in plant yield in terms of seed weight. Measurement of Na(+) ions in different parts of the plant showed higher accumulation in the old leaves and negligible accumulation in seeds of T(1) transgenic lines as compared with the WT plants. The possible mechanism of salinity tolerance is discussed. Overexpression of PDH45 provides a possible example of the exploitation of DNA/RNA unwinding pathways for engineering salinity tolerance without affecting yield in crop plants.

  17. 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.

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

    SciTech Connect

    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.; Sorce, A.; Sorce, C.; Sangster, T. C.; Weiner, D.

    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 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. Furthermore, the measured instrument response includes contributions from the scintillator rise time, optical relay, and streak camera.

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

    PubMed

    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.

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

    DOE PAGES

    Stoeckl, C.; Boni, R.; Ehrne, F.; ...

    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

  1. GPCR engineering yields high-resolution structural insights into beta2-adrenergic receptor function.

    PubMed

    Rosenbaum, Daniel M; Cherezov, Vadim; Hanson, Michael A; Rasmussen, Søren G F; Thian, Foon Sun; Kobilka, Tong Sun; Choi, Hee-Jung; Yao, Xiao-Jie; Weis, William I; Stevens, Raymond C; Kobilka, Brian K

    2007-11-23

    The beta2-adrenergic receptor (beta2AR) is a well-studied prototype for heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptors (GPCRs) that respond to diffusible hormones and neurotransmitters. To overcome the structural flexibility of the beta2AR and to facilitate its crystallization, we engineered a beta2AR fusion protein in which T4 lysozyme (T4L) replaces most of the third intracellular loop of the GPCR ("beta2AR-T4L") and showed that this protein retains near-native pharmacologic properties. Analysis of adrenergic receptor ligand-binding mutants within the context of the reported high-resolution structure of beta2AR-T4L provides insights into inverse-agonist binding and the structural changes required to accommodate catecholamine agonists. Amino acids known to regulate receptor function are linked through packing interactions and a network of hydrogen bonds, suggesting a conformational pathway from the ligand-binding pocket to regions that interact with G proteins.

  2. Morphological characterization and assessment of genetic variability, character association, and divergence in soybean mutants.

    PubMed

    Malek, M A; Rafii, Mohd Y; Shahida Sharmin Afroz, Most; Nath, Ujjal Kumar; Mondal, M Monjurul Alam

    2014-01-01

    Genetic diversity is important for crop improvement. An experiment was conducted during 2011 to study genetic variability, character association, and genetic diversity among 27 soybean mutants and four mother genotypes. Analysis of variance revealed significant differences among the mutants and mothers for nine morphological traits. Eighteen mutants performed superiorly to their mothers in respect to seed yield and some morphological traits including yield attributes. Narrow differences between phenotypic and genotypic coefficients of variation (PCV and GCV) for most of the characters revealed less environmental influence on their expression. High values of heritability and genetic advance with high GCV for branch number, plant height, pod number, and seed weight can be considered as favorable attributes for soybean improvement through phenotypic selection and high expected genetic gain can be achieved. Pod and seed number and maturity period appeared to be the first order traits for higher yield and priority should be given in selection due to their strong associations and high magnitudes of direct effects on yield. Cluster analysis grouped 31 genotypes into five groups at the coefficient value of 235. The mutants/genotypes from cluster I and cluster II could be used for hybridization program with the mutants of clusters IV and V in order to develop high yielding mutant-derived soybean varieties for further improvement.

  3. Morphological Characterization and Assessment of Genetic Variability, Character Association, and Divergence in Soybean Mutants

    PubMed Central

    Malek, M. A.; Rafii, Mohd Y.; Shahida Sharmin Afroz, Most.; Nath, Ujjal Kumar; Mondal, M. Monjurul Alam

    2014-01-01

    Genetic diversity is important for crop improvement. An experiment was conducted during 2011 to study genetic variability, character association, and genetic diversity among 27 soybean mutants and four mother genotypes. Analysis of variance revealed significant differences among the mutants and mothers for nine morphological traits. Eighteen mutants performed superiorly to their mothers in respect to seed yield and some morphological traits including yield attributes. Narrow differences between phenotypic and genotypic coefficients of variation (PCV and GCV) for most of the characters revealed less environmental influence on their expression. High values of heritability and genetic advance with high GCV for branch number, plant height, pod number, and seed weight can be considered as favorable attributes for soybean improvement through phenotypic selection and high expected genetic gain can be achieved. Pod and seed number and maturity period appeared to be the first order traits for higher yield and priority should be given in selection due to their strong associations and high magnitudes of direct effects on yield. Cluster analysis grouped 31 genotypes into five groups at the coefficient value of 235. The mutants/genotypes from cluster I and cluster II could be used for hybridization program with the mutants of clusters IV and V in order to develop high yielding mutant-derived soybean varieties for further improvement. PMID:25197722

  4. High yield sample preconcentration using a highly ion-conductive charge-selective polymer.

    PubMed

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

    2010-07-15

    The development and analysis of a microfluidic sample preconcentration system using a highly ion-conductive charge-selective polymer [poly-AMPS (2-acrylamido-2-methyl-1-propanesulfonic acid)] 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 microM (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.

  5. 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

  6. Chemical and genetic diversity of high-seed-yield sorghum (Sorghum bicolor M.) germplasms.

    PubMed

    Ryu, J; Im, S B; Kwon, S J; Ahn, J W; Jeong, S W; Kang, S Y

    2016-09-02

    This study evaluated the chemical and genetic diversity of high-seed-yield sorghum germplasms from Korea, the United States, and South Africa. We identified significant differences in the chemical contents of whole plants at the heading stage in all cultivars, including differences in crude protein, fat, fiber, ash, neutral detergent fiber, acid detergent fiber, mineral, and fatty acid contents. Our results suggest that Banwoldang is the most appropriate cultivar for roughage because of its high protein yield. We identified significant differences in the tannin, flavonoid, amylose, mineral, crude fat, fatty acid, and 3-deoxyanthocyanin contents in the whole grain from all cultivars, but not in the mineral or crude fat contents. Tannin levels were generally low. IS645 contained the highest levels of flavonoids and linolenic acid compounds, and Moktak had the highest amylose and deoxyanthocyanidin content in the grain. To assess genetic diversity, we used 10 simple sequence repeat (SSR) primer sets to identify 38 alleles with 3-8 alleles per locus. Based on phylogenetic analysis of the SSR markers, the sorghum cultivars were divided into three major groups. Comparison of clusters based on chemical compositions with those based on SSRs showed that the groups formed by the three native Korean cultivars clustered similarly in molecular dendrograms. Association analysis was conducted for the 10 SSR marker; 48 chemical and growth traits were present for two marker traits (seed color and whole plant fatty acid content) with significant marker-trait associations. These markers could be used to select sorghum cultivars for breeding programs.

  7. High yield ICF target design for a Z-pinch driven hohlraum

    SciTech Connect

    Bailey, D.S.; Hammer, J.H.; Lindl, J.D.; Rambo, P.W.; Tabak, M.; Toor, A.; Wilks, S.C.; Zimmerman, G.B

    1998-11-13

    We describe calculations for a high yield inertial fusion design, employing indirect drive with a double-ended z-pinch-driven hohlraum radiation source. A high current ({approximately}60 MA) accelerator implodes z-pinches within an enclosing hohlraum. Radial spoke arrays and shine shields isolate the capsule from the pinch plasma, magnetic field and direct x-ray shine. Our approach places minimal requirements on z-pinch uniformity and stability, usually problematic due to magneto-Rayleigh Taylor (MRT) instability. Large inhomogeneities of the pinch and spoke array may be present, but the hohlraum adequately smooths the radiation field at the capsule. Simultaneity and reproducibility of the pinch x-ray output to better than 7% are required, however, for good symmetry. Recent experiments suggest a pulse shaping technique, through implosion of a multishell z-pinch. X-ray bursts are calculated and observed to occur at each shell collision. A capsule absorbing 1 MJ of x-rays at a peak drive temperature of 210 eV is found to have adequate stability and to produce 400 MJ of yield.

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

    PubMed

    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

    2015-09-01

    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.

  9. Functionalization of quinoxalines by using TMP bases: preparation of tetracyclic heterocycles with high photoluminescene quantum yields.

    PubMed

    Nafe, Julia; Herbert, Simon; Auras, Florian; Karaghiosoff, Konstantin; Bein, Thomas; Knochel, Paul

    2015-01-12

    Tetracyclic heterocycles that exhibit high photoluminescence quantum yields were synthesized by anellation reactions of mono-, di-, and trifunctionalized 2,3-dichloroquinoxalines. Thus, treatment of 2,3-dichloroquinoxaline with TMPLi (TMP = 2,2,6,6-tetramethylpiperidyl) allows a regioselective lithiation in position 5. Quenching with various electrophiles (iodine, (BrCl2 C)2 , allylic bromide, acid chloride, aryl iodide) leads to 5-functionalized 2,3-dichloroquinoxalines. Further functionalization in positions 6 and 8 can be achieved by using TMPLi or TMPMgCl⋅LiCl furnishing a range of new di- and tri-functionalized 2,3-dichloroquinoxalines. The chlorine atoms are readily substituted by anellation with 1,2-diphenols or 1,2-dithiophenols leading to a series of new tetracyclic compounds. These materials exhibit strong, tunable optical absorption and emission in the blue and green spectral region. The substituted O-heterocyclic compounds exhibit particularly high photoluminescence quantum yields of up to 90%, which renders them interesting candidates for fluorescence imaging applications.

  10. 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

  11. Mechanically activated catalyst mixing for high-yield boron nitride nanotube growth

    NASA Astrophysics Data System (ADS)

    Li, Ling; Li, Lu Hua; Chen, Ying; Dai, Xiujuan J.; Xing, Tan; Petravic, Mladen; Liu, Xiaowei

    2012-07-01

    Boron nitride nanotubes (BNNTs) have many fascinating properties and a wide range of applications. An improved ball milling method has been developed for high-yield BNNT synthesis, in which metal nitrate, such as Fe(NO3)3, and amorphous boron powder are milled together to prepare a more effective precursor. The heating of the precursor in nitrogen-containing gas produces a high density of BNNTs with controlled structures. The chemical bonding and structure of the synthesized BNNTs are precisely probed by near-edge X-ray absorption fine structure spectroscopy. The higher efficiency of the precursor containing milling-activated catalyst is revealed by thermogravimetric analyses. Detailed X-ray diffraction and X-ray photoelectron spectroscopy investigations disclose that during ball milling the Fe(NO3)3 decomposes to Fe which greatly accelerates the nitriding reaction and therefore increases the yield of BNNTs. This improved synthesis method brings the large-scale production and application of BNNTs one step closer.

  12. Mechanically activated catalyst mixing for high-yield boron nitride nanotube growth.

    PubMed

    Li, Ling; Li, Lu Hua; Chen, Ying; Dai, Xiujuan J; Xing, Tan; Petravic, Mladen; Liu, Xiaowei

    2012-07-24

    Boron nitride nanotubes (BNNTs) have many fascinating properties and a wide range of applications. An improved ball milling method has been developed for high-yield BNNT synthesis, in which metal nitrate, such as Fe(NO3)3, and amorphous boron powder are milled together to prepare a more effective precursor. The heating of the precursor in nitrogen-containing gas produces a high density of BNNTs with controlled structures. The chemical bonding and structure of the synthesized BNNTs are precisely probed by near-edge X-ray absorption fine structure spectroscopy. The higher efficiency of the precursor containing milling-activated catalyst is revealed by thermogravimetric analyses. Detailed X-ray diffraction and X-ray photoelectron spectroscopy investigations disclose that during ball milling the Fe(NO3)3 decomposes to Fe which greatly accelerates the nitriding reaction and therefore increases the yield of BNNTs. This improved synthesis method brings the large-scale production and application of BNNTs one step closer.

  13. 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.

  14. Lipid metabolites in seeds of diverse Gossypium accessions: Molecular identification of a high oleic mutant allele

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The domestication and breeding of cotton for elite, high-fiber cultivars has led to reduced genetic variation of seed constituents within currently cultivated upland Cotton genotypes. However, a recent screen of the genetically diverse U.S. National Cotton Germplasm Collection identified Gossypium ...

  15. Breeding of a sake yeast mutant with enhanced ethyl caproate productivity in sake brewing using rice milled at a high polishing ratio.

    PubMed

    Takahashi, Toshinari; Ohara, Yusuke; Sueno, Kazuo

    2017-03-09

    Sake yeast produces a fruity flavor known as ginjo-ko-which is mainly attributable to ethyl caproate and isoamyl acetate-during fermentation in sake brewing. The production of these flavor components is inhibited by unsaturated fatty acids derived from the outer layer of rice as raw material. We isolated three mutants (hec2, hec3, and hec6) with enhanced ethyl caproate productivity in sake brewing using rice milled at a high polishing ratio from a cerulenin-resistant mutant derived from the hia1 strain, which shows enhanced isoamyl acetate productivity. The hec2 mutant had the homozygous FAS2 mutation Gly1250Ser, which is known to confer high ethyl caproate productivity. When the homozygous FAS2 mutation Gly1250Ser was introduced into strain hia1, ethyl caproate productivity was increased but neither this nor intracellular caproic acid content approached the levels observed in the hec2 mutant, indicating that a novel mutation was responsible for the high ethyl caproate productivity. We also found that the expression of EEB1 encoding acyl-coenzyme A: ethanol O-acyltransferase (AEATase) and enzymatic activity were increased in the hec2 mutant. These results suggest that the upregulation of EEB1 expression and AEATase activity may also have contributed to the enhancement of ethyl caproate synthesis from ethanol and caproyl-CoA. Our findings are useful for the brewing of sake with improved flavor due to high levels of isoamyl acetate and ethyl caproate.

  16. Stability and activity of Dictyoglomus thermophilum GH11 xylanase and its disulphide mutant at high pressure and temperature.

    PubMed

    Li, He; Voutilainen, Sanni; Ojamo, Heikki; Turunen, Ossi

    2015-03-01

    The functional properties of extremophilic Dictyoglomus thermophilum xylanase (XYNB) and the N-terminal disulphide-bridge mutant (XYNB-DS) were studied at high pressure and temperature. The enzymes were quite stable even at the pressure of 500MPa at 80°C. The half-life of inactivation in these conditions was over 30h. The inactivation at 80°C in atmospheric pressure was only 3-times slower. The increase of pressure up to 500MPa at 80°C decreased only slightly the enzyme's stability, whereas in 500MPa the increase of temperature from 22 to 80°C decreased significantly more the enzyme's stability. While the high temperature (80-100°C) decreased the enzyme reaction with short xylooligosaccharides (xylotetraose and xylotriose), the high pressure (100-300MPa) had an opposite effect. The temperature of 100°C strongly increased the Km but did not affect the kcat to the same extent, thus indicating that the interaction of the substrate with the active site suffers before the catalytic reaction begins to decrease as the temperature rises. Circular dichroism spectroscopy showed the high structural stability of XYNB and XYNB-DS at 93°C.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. A Microfluidic Platform for High-Throughput Screening of Small Mutant Libraries.

    PubMed

    Lim, Ji Won; Shin, Kwang Soo; Moon, Jaemin; Lee, Sung Kuk; Kim, Taesung

    2016-05-17

    The screening and isolation of target microorganisms from mutated recombinant libraries are crucial for the advancement of synthetic biology and metabolic engineering. However, conventional screening tools present several limitations in throughput, cost, and labor. Herein, we describe a novel microfluidic high-throughput screening (HTS) platform with several advantages. The platform utilizes a fluid array to compartmentalize bacterial cells in well-ordered separated microwells and allows long-term cell culture with high throughput. The platform enables the extraction of selected target cells from the fluid array for additional culture and postanalysis by using a capillary-driven sample relocation method. To confirm the feasibility of the platform, we demonstrated two different types of HTS methods based on the levels of reporter gene expression and cellular growth rate difference. For the reporter gene-based HTS, a spike recovery approach was taken to demonstrate that target cells are successfully screened out from a mixture containing nontarget cells by repeating the culture and extraction processes. Additionally, the same platform allowed us to screen and sort target cells according to their cellular growth rate difference, which seems hard in conventional screening methods. Hence, the platform could be used for various microbiological assays, including the detection of cell-excreted metabolites, microbial biosensors, and other HTS systems.

  2. 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.

  3. 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.

  4. Engineering a high-yield glutathione strain of Hansenula polymorpha using ion beam implantation.

    PubMed

    Qian, Weidong; Fu, Yunfang; Cai, Changlong

    2013-01-01

    To generate an industrial strain of Hansenula polymorpha capable of yielding greater levels of glutathione (GSH), wild strain H. polymorpha DL-1 cells were mutated using a nitrogen ion beam, a novel mutagen. At an energy level of 20 keV and dose of 2.13 × 10(16) ions/cm(2), H. polymorpha strain 28 (HP28) with a high-yield of GSH was screened. HP28 intracellular GSH levels reached 337.16 mg/L by ion beam implantation, 1.56 times greater than that of the wild type strain when the fermentation time was shortened from 48 hr to 42 hr, greatly improving efficiency and reducing the cost of industrial-scale production. The enhanced efficiency of HP28 is promising for GSH production from lignocellulosic materials. Therefore, the ion beam implantation would be a cost-effective alternative to the conventional mutation method for engineering yeast and improving its utility.

  5. Prediction of yield stress and Charpy transition temperature in highly neutron irradiated ferritic steels

    NASA Astrophysics Data System (ADS)

    Windsor, Colin; Cottrell, Geoff; Kemp, Richard

    2010-07-01

    Recent predictions have been made of metallurgical properties of low-activation ferritic/martensitic steels alloys at the high irradiation levels (displacements per atom or dpa) needed for a fusion power plant as based on measurements at low irradiation levels where more data are available. These predictions have been published for the yield stress and for the Charpy ductile to brittle transition temperature shift. The neural network model predictions use training data up to a certain dpa level to predict metallurgical properties above this level. This 'extrapolation' mode of neural networks is explored in some detail. Our studies revealed an increasing accuracy of predictions as the test dpa level is increased for both yield stress and Charpy shift predictions. This result suggests that a model exists for these metallurgical properties as a function of dpa level which becomes more accurate as the available irradiation range in the training data is increased. The explanation suggested is that the metallurgical annealing, which occurs as the irradiation level is increased, simplifies the microstructure and makes prediction more reliable.

  6. Mixed models for selection of Jatropha progenies with high adaptability and yield stability in Brazilian regions.

    PubMed

    Teodoro, P E; Bhering, L L; Costa, R D; Rocha, R B; Laviola, B G

    2016-08-19

    The aim of this study was to estimate genetic parameters via mixed models and simultaneously to select Jatropha progenies grown in three regions of Brazil that meet high adaptability and stability. From a previous phenotypic selection, three progeny tests were installed in 2008 in the municipalities of Planaltina-DF (Midwest), Nova Porteirinha-MG (Southeast), and Pelotas-RS (South). We evaluated 18 families of half-sib in a randomized block design with three replications. Genetic parameters were estimated using restricted maximum likelihood/best linear unbiased prediction. Selection was based on the harmonic mean of the relative performance of genetic values method in three strategies considering: 1) performance in each environment (with interaction effect); 2) performance in each environment (with interaction effect); and 3) simultaneous selection for grain yield, stability and adaptability. Accuracy obtained (91%) reveals excellent experimental quality and consequently safety and credibility in the selection of superior progenies for grain yield. The gain with the selection of the best five progenies was more than 20%, regardless of the selection strategy. Thus, based on the three selection strategies used in this study, the progenies 4, 11, and 3 (selected in all environments and the mean environment and by adaptability and phenotypic stability methods) are the most suitable for growing in the three regions evaluated.

  7. High yield bio-oil production from fast pyrolysis by metabolic controlling of Chlorella protothecoides.

    PubMed

    Miao, Xiaoling; Wu, Qingyu

    2004-05-13

    The use of renewable energy sources is becoming increasingly necessary to mitigate global warming. Recently much research has been focused on identifying suitable biomass species, which can provide high-energy outputs, to replace conventional fossil fuels. This paper reports an approach for increasing the yield of bio-oil production from fast pyrolysis after manipulating the metabolic pathway in microalgae through heterotrophic growth. The yield of bio-oil (57.9%) produced from heterotrophic Chlorella protothecoides cells was 3.4 times higher than from autotrophic cells by fast pyrolysis. The bio-oil was characterized by a much lower oxygen content, with a higher heating value (41 MJ kg(-1)), a lower density (0.92 kg l(-1)), and lower viscosity (0.02 Pas) compared to those of bio-oil from autotrophic cells and wood. These properties are comparable to fossil oil. The research could contribute to the creation of a system to produce energy from microalgae, and also could have great commercial potential for liquid fuel production.

  8. Solution-processable SWNT transistors: Toward high performance and full semiconducting device yield

    NASA Astrophysics Data System (ADS)

    Zhang, Wenjing; Lee, Chun Wei; Zhao, Jianwen; Li, Lain-Jong

    2010-03-01

    The major hurdle to scale up SWNT network field-effect transistors (FETs) is the difficulty in obtaining high mobility and full semiconductor device yield due to the co-existence of metallic and semiconducting tubes in fabricated networks. We demonstrate that the radical initiator 1,1'-azobis(cyanocyclohexane) (ACN) allows for release of radicals which preferentially react with small and metallic SWNTs. By applying this reaction to CoMoCat SWNTs, metallic tubes are electrically suppressed and semiconducting thin-film transistors can be readily made with almost full semiconductor device yield. The effective mobility can be raised to ˜10 cm2/V.s with increasing the network thickness to ˜20 nm while keeping the on-off ratio higher than 10000. Moreover, top-gated devices with polymer-ionic liquid mixture as the dielectrics are demonstrated to be almost hysteresis-free and with low threshold voltage, promising applications in low-cost printable electronics.

  9. 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.

  10. Photoisomerization dynamics of a rhodopsin-based molecule (potential molecular switch) with high quantum yields

    NASA Astrophysics Data System (ADS)

    Allen, Roland; Jiang, Chen-Wei; Zhang, Xiu-Xing; Fang, Ai-Ping; Li, Hong-Rong; Xie, Rui-Hua; Li, Fu-Li

    2015-03-01

    It is worthwhile to explore the detailed reaction dynamics of various candidates for molecular switches, in order to understand, e.g., the differences in quantum yields and switching times. Here we report density-functional-based simulations for the rhodopsin-based molecule 4-[4-Methylbenzylidene]-5-p-tolyl-3,4-dihydro-2H-pyrrole (MDP), synthesized by Sampedro et al. We find that the photoisomerization quantum yields are remarkably high: 82% for cis-to-trans, and 68% for trans-to-cis. The lifetimes of the S1 excited state in cis-MDP in our calculations are in the range of 900-1800 fs, with a mean value of 1270 fs, while the range of times required for full cis-to-trans isomerization are 1100-2000 fs, with a mean value of 1530 fs. In trans-MDP, the calculated S1 excited state lifetimes are 860-2140 fs, with a mean value of 1330 fs, and with the full trans-to-cis isomerization completed about 200 fs later. In both cases, the dominant reaction mechanism is rotation around the central C =C bond (connected to the pyrroline ring), and de-excitation occurs at an avoided crossing between the ground state and the lowest singlet state, near the midpoint of the rotational pathway. Research Fund for the Doctoral Program of Higher Education of China; Fundamental Research Funds for the Central Universities; Robert A. Welch Foundation; National Natural Science Foundation of China.

  11. Photoisomerization dynamics of a rhodopsin-based molecule (potential molecular switch) with high quantum yields

    NASA Astrophysics Data System (ADS)

    Jiang, Chen-Wei; Zhang, Xiu-Xing; Fang, Ai-Ping; Li, Hong-Rong; Xie, Rui-Hua; Li, Fu-Li; Allen, Roland E.

    2015-02-01

    It is worthwhile to explore the detailed reaction dynamics of various candidates for molecular switches, in order to understand, e.g., the differences in quantum yields and switching times. Here we report density-functional-based simulations for the rhodopsin-based molecule 4-[4-methylbenzylidene]-5-p-tolyl-3,4-dihydro-2H-pyrrole (MDP), synthesized by Sampedro et al We find that the photoisomerization quantum yields are remarkably high: 82% for cis-to-trans, and 68% for trans-to-cis. The lifetimes of the S1 excited state in cis-MDP in our calculations are in the range of 900-1800 fs, with a mean value of 1270 fs, while the range of times required for full cis-to-trans isomerization are 1100-2000 fs, with a mean value of 1530 fs. In trans-MDP, the calculated S1 excited state lifetimes are 860-2140 fs, with a mean value of 1330 fs, and with the full trans-to-cis isomerization completed about 200 fs later. In both cases, the dominant reaction mechanism is rotation around the central C=C bond (connected to the pyrroline ring), and de-excitation occurs at an avoided crossing between the ground state and the lowest singlet state, near the midpoint of the rotational pathway. Perhaps remarkably, but apparently because of electrostatic repulsion, the direction of rotation is the same for both reactions.

  12. 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.

  13. 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).

  14. Enhanced ethanol production at commercial scale from molasses using high gravity technology by mutant S. cerevisiae.

    PubMed

    Arshad, Muhammad; Hussain, Tariq; Iqbal, Munawar; Abbas, Mazhar

    2017-02-16

    Very high gravity (VHG) technology was employed on industrial scale to produce ethanol from molasses (fermented) as well as by-products formation estimation. The effect of different Brix° (32, 36 and 40) air-flow rates (0.00, 0.20, 0.40, and 0.60vvm) was studied on ethanol production. The maximum ethanol production was recorded to be 12.2% (v/v) at 40 Brix° with 0.2vvm air-flow rate. At optimum level aeration and 40 Brix° VHG, the residual sugar level was recorded in the range of 12.5-18.5g/L, whereas the viable cell count remained constant up to 50h of fermentation and dry matter production increased with fermentation time. Both water and steam consumption reduced significantly under optimum conditions of Brix° and aeration rate with compromising the ethanol production. Results revealed VHG with continuous air flow is viable technique to reduce the ethanol production cost form molasses at commercial scale.

  15. 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.

  16. 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

    2015-08-07

    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.

  17. High-yield fermentative preparation of tetramethylpyrazine by Bacillus sp. using an endogenous precursor approach.

    PubMed

    Zhu, Bing-Feng; Xu, Yan; Fan, Wen-Lai

    2010-02-01

    A spore-forming Bacillus sp. was isolated from a high-temperature Daqu, a starter culture of Chinese Maotai-flavor liquor, using an endogenous precursor screening strategy. The Bacillus sp. was capable of producing a high level of 2,3,5,6-tetramethylpyrazine (TTMP) via a precursor of 3-hydroxy-2-butanone (HB). The strain was characterized as Bacillus subtilis based on morphological, physiological, and biochemical properties as well as on partial 16S rRNA gene sequences. Different carbon and nitrogen sources as well as fermentation conditions were investigated. Optimization tests showed that oxygen supply and fermentation temperature were the most important parameters determining the production process. The production of >4.08 g/l TTMP was achieved together with a high level of endogenous precursor HB accumulation (>20 g/l) in both flask and fermentor cultures when the optimized medium and cultivation conditions were applied. Our data demonstrates the effectiveness of the endogenous precursor strategy for screening microorganisms that produce flavor compounds with structure-related precursors. The high yield of TTMP and the inexpensiveness of the agro-industrial product used as the substrate (soybean meal) indicate the potential of this process for industrial application.

  18. DEVELOPMENT OF NOVEL, ALTERNATIVE, FACILE, ECOFRIENDLY, HIGH YIELD SYNTHETIC PROCESS FOR PRAZOSIN

    PubMed Central

    Patil, D. A.; Jain, K. S.; Deodhar, M. N.; Patil, P. O.; Patil, G. B.; Patil, D. D.

    2010-01-01

    Industrial chemistry in the new millennium is widely adopting the concept of “Green chemistry” to meet the fundamental scientific challenges. Antihypertensive drugs include several of the most widely prescribed drugs like diuretics, beta-blockers, ACE inhibitors, calcium channel blockers, and α-1 adrenoreceptor blockers. The discovery of prazosin, with very high index of α1/α2 affinity has triggered off a renaissance of interest in α1-adrenoceptor antagonist drugs for treatment of hypertension. The three reported routes for synthesis and manufacture of the α-adrenoceptor antagonist- prazosin had some disadvantages. In present study we had developed new methods for the synthesis of prazosin by using microwave. The most important aspect is the overall yield of this process was ~25 % higher than the other reported methods excluding the use of banned substances PMID:24825992

  19. Film quantum yields of EUV and ultra-high PAG photoresists

    NASA Astrophysics Data System (ADS)

    Hassanein, Elsayed; Higgins, Craig; Naulleau, Patrick; Matyi, Richard; Gallatin, Gregg; Denbeaux, Gregory; Antohe, Alin; Thackeray, 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

    2008-03-01

    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 of PAG show divergent mechanisms for production of photoacids beyond PAG concentrations of 0.35 moles/liter. The FQY of sulfonium PAGs level off, whereas resists prepared with iodonium PAG show FQYs that increase beyond PAG concentrations of 0.35 moles/liter, reaching record highs of 8-13 acids generated/EUV photons absorbed.

  20. High-yield synthesis of silicon carbide nanowires by solar and lamp ablation

    NASA Astrophysics Data System (ADS)

    Lu, Hai-bo; Chan, Benjamin C. Y.; Wang, Xiaolin; Tong Chua, Hui; Raston, Colin L.; Albu-Yaron, Ana; Levy, Moshe; Popowitz-Biro, Ronit; Tenne, Reshef; Feuermann, Daniel; Gordon, Jeffrey M.

    2013-08-01

    We report a reasonably high yield (∼50%) synthesis of silicon carbide (SiC) nanowires from silicon oxides and carbon in vacuum, by novel solar and lamp photothermal ablation methods that obviate the need for catalysis, and allow relatively short reaction times (∼10 min) in a nominally one-step process that does not involve toxic reagents. The one-dimensional core/shell β-SiC/SiOx nanostructures—characterized by SEM, TEM, HRTEM, SAED, XRD and EDS—are typically several microns long, with core and outer diameters of about 10 and 30 nm, respectively. HRTEM revealed additional distinctive nanoscale structures that also shed light on the formation pathways.

  1. 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.

  2. 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.

  3. High yield wind energy resources in the mid-Hudson River basin. Final report

    SciTech Connect

    Sforza, P.M.; Smorto, M.J.; Bailey, B.; Wheeler, R.

    1983-12-01

    An evaluation of high-yield wind energy resources in the franchise area of the Central Hudson Gas and Electric Corporation is presented. A site screening process was used to select seven sites for field tests. Wind data taken at those sites for three months were used as the basis to select four sites for an additional testing period of nine months. A computer simulation was conducted on the performance of eight wind turbines (55 kW to 7350 kW) at the four sites based on the one year data recorded; the results of the computer runs are described. Also, an economic analysis of the operation of the eight wind turbines at the windiest site is presented. Recommendations for wind energy development in the region are discussed.

  4. HYPERSENSITIVE TO HIGH LIGHT1 Interacts with LOW QUANTUM YIELD OF PHOTOSYSTEM II1 and Functions in Protection of Photosystem II from Photodamage in Arabidopsis[C][W][OPEN

    PubMed Central

    Jin, Honglei; Liu, Bing; Luo, Lujun; Feng, Dongru; Wang, Peng; Liu, Jun; Da, Qingen; He, Yanming; Qi, Kangbiao; Wang, Jinfa; Wang, Hong-Bin

    2014-01-01

    Under high-irradiance conditions, plants must efficiently protect photosystem II (PSII) from damage. In this study, we demonstrate that the chloroplast protein HYPERSENSITIVE TO HIGH LIGHT1 (HHL1) is expressed in response to high light and functions in protecting PSII against photodamage. Arabidopsis thaliana hhl1 mutants show hypersensitivity to high light, drastically decreased PSII photosynthetic activity, higher nonphotochemical quenching activity, a faster xanthophyll cycle, and increased accumulation of reactive oxygen species following high-light exposure. Moreover, HHL1 deficiency accelerated the degradation of PSII core subunits under high light, decreasing the accumulation of PSII core subunits and PSII–light-harvesting complex II supercomplex. HHL1 primarily localizes in the stroma-exposed thylakoid membranes and associates with the PSII core monomer complex through direct interaction with PSII core proteins CP43 and CP47. Interestingly, HHL1 also directly interacts, in vivo and in vitro, with LOW QUANTUM YIELD OF PHOTOSYSTEM II1 (LQY1), which functions in the repair and reassembly of PSII. Furthermore, the hhl1 lqy1 double mutants show increased photosensitivity compared with single mutants. Taken together, these results suggest that HHL1 forms a complex with LQY1 and participates in photodamage repair of PSII under high light. PMID:24632535

  5. 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

  6. A high loading overland flow system: Impacts on soil characteristics, grass constituents, yields and nutrient removal.

    PubMed

    Wen, C G; Chen, T H; Hsu, F H; Lu, C H; Lin, J B; Chang, C H; Chang, S P; Lee, C S

    2007-04-01

    The objectives of this paper are to determine effects of different grass species and their harvests on pollutant removal, elucidate impacts on soil characteristics and grass constituents, observe grass yield and quantify nutrient uptake by vegetation in an overland flow system (OLFS). Polluted creek water was applied to eight channels in the OLFS, which were planted with Paragrass, Nilegrass, Cattail, and Vetiver, with each two channels being randomly planted with a given grass species. The grass in one channel was harvested while that in the other channel was not. At a high rate of 27.8 m d(-1) hydraulic loading, the removal efficiencies of conventional pollutants such as BOD, COD, suspended solids (SS), and total coliforms in wastewater are not affected by the type of the grasses species, but those of nitrogen and phosphorus are affected by different species. Overall average removal efficiencies of BOD, COD, SS, ammonia, total nitrogen, total phosphorus and total coliforms through the OLFS are 42%, 48%, 78%, 47%, 40%, 33% and 89%, respectively. The concentration of nitrate, however, increases due to nitrification. Soil characteristics in OLFS have been changed significantly; specific conductivity, organic matter, exchangeable magnesium, extractable copper and zinc in soils all increase with time while pHs decrease. During the winter season, there is a significant accumulation of nitrate in grass with the subsequent reduction during the active growing season (Spring). The contents of nitrate and phosphorus in grass tissue are higher than those of grass in general pastureland, probably due to nutrient luxury uptake by grass. The overall grass yield, growth rate and nutrient uptake are quantified and implication of such high rate OLFS discussed.

  7. Highly efficient gene targeting in Penicillium chrysogenum using the bi-partite approach in deltalig4 or deltaku70 mutants.

    PubMed

    de Boer, Paulo; Bastiaans, Jeroen; Touw, Hesselien; Kerkman, Richard; Bronkhof, Jurian; van den Berg, Marco; Offringa, Remko

    2010-10-01

    Inactivating the non-homologous end-joining (NHEJ) pathway is a well established method to increase gene targeting (GT) efficiencies in filamentous fungi. In this study we have compared the effect of inactivating the NHEJ genes ku70 or lig4 on GT in the industrial penicillin producer Penicillium chrysogenum. Deletion of both genes resulted in strongly increased GT efficiencies at three different loci but not higher than 70%, implying that other, yet uncharacterized, recombination pathways are still active causing a part of the DNA to be integrated via non-homologous recombination. To further increase the GT efficiency we applied the bi-partite approach, in which the DNA fragment for integration was split in two non-functional overlapping parts that via homologous recombination invivo can form a functional selection marker. The combined NHEJ mutant and bi-partite approach further increased GT frequencies up to approximately 90%, which will enable the efficient high throughput engineering of the P. chrysogenum genome. We expect that this combined approach will function with similar high efficiencies in other filamentous fungi.

  8. 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

  9. A High Yield and Cost-efficient Expression System of Human Granzymes in Mammalian Cells.

    PubMed

    Dotiwala, Farokh; Fellay, Isabelle; Filgueira, Luis; Martinvalet, Denis; Lieberman, Judy; Walch, Michael

    2015-06-10

    When cytotoxic T lymphocytes (CTL) or natural killer (NK) cells recognize tumor cells or cells infected with intracellular pathogens, they release their cytotoxic granule content to eliminate the target cells and the intracellular pathogen. Death of the host cells and intracellular pathogens is triggered by the granule serine proteases, granzymes (Gzms), delivered into the host cell cytosol by the pore forming protein perforin (PFN) and into bacterial pathogens by the prokaryotic membrane disrupting protein granulysin (GNLY). To investigate the molecular mechanisms of target cell death mediated by the Gzms in experimental in-vitro settings, protein expression and purification systems that produce high amounts of active enzymes are necessary. Mammalian secreted protein expression systems imply the potential to produce correctly folded, fully functional protein that bears posttranslational modification, such as glycosylation. Therefore, we used a cost-efficient calcium precipitation method for transient transfection of HEK293T cells with human Gzms cloned into the expression plasmid pHLsec. Gzm purification from the culture supernatant was achieved by immobilized nickel affinity chromatography using the C-terminal polyhistidine tag provided by the vector. The insertion of an enterokinase site at the N-terminus of the protein allowed the generation of active protease that was finally purified by cation exchange chromatography. The system was tested by producing high levels of cytotoxic human Gzm A, B and M and should be capable to produce virtually every enzyme in the human body in high yields.

  10. High-yield Synthesis of Multiwalled Carbon Nanotube by Mechanothermal Method

    NASA Astrophysics Data System (ADS)

    Manafi, S. A.; Amin, M. H.; Rahimipour, M. R.; Salahi, E.; Kazemzadeh, A.

    2009-04-01

    This study reports on the mechanothermal synthesis of multiwalled carbon nanotube (MWCNTs) from elemental graphite powder. Initially, high ultra-active graphite powder can be obtained by mechanical milling under argon atmosphere. Finally, the mechanical activation product is heat-treated at 1350°C for 2-4 h under argon gas flow. After heat-treatment, active graphite powders were successfully changed into MWCNTs with high purity. The XRD analyses showed that in the duration 150 h of milling, all the raw materials were changed to the desired materials. From the broadening of the diffraction lines in the XRD patterns, it was concluded that the graphite crystallites were nanosized, and raising the milling duration resulted in the fineness of the particles and the increase of the strain. The structure and morphology of MWCNTs were investigated using scanning electron microscopy (SEM) and high-resolution transmission electron microscopy (HRTEM). The yield of MWCNTs was estimated through SEM and TEM observations of the as-prepared samples was to be about 90%. Indeed, mechanothermal method is of interest for fundamental understanding and improvement of commercial synthesis of carbon nanotubes (CNTs). As a matter of fact, the method of mechanothermal guarantees the production of MWCNTs suitable for different applications.

  11. High-yield Synthesis of Multiwalled Carbon Nanotube by Mechanothermal Method.

    PubMed

    Manafi, S A; Amin, M H; Rahimipour, M R; Salahi, E; Kazemzadeh, A

    2009-01-22

    This study reports on the mechanothermal synthesis of multiwalled carbon nanotube (MWCNTs) from elemental graphite powder. Initially, high ultra-active graphite powder can be obtained by mechanical milling under argon atmosphere. Finally, the mechanical activation product is heat-treated at 1350 degrees C for 2-4 h under argon gas flow. After heat-treatment, active graphite powders were successfully changed into MWCNTs with high purity. The XRD analyses showed that in the duration 150 h of milling, all the raw materials were changed to the desired materials. From the broadening of the diffraction lines in the XRD patterns, it was concluded that the graphite crystallites were nanosized, and raising the milling duration resulted in the fineness of the particles and the increase of the strain. The structure and morphology of MWCNTs were investigated using scanning electron microscopy (SEM) and high-resolution transmission electron microscopy (HRTEM). The yield of MWCNTs was estimated through SEM and TEM observations of the as-prepared samples was to be about 90%. Indeed, mechanothermal method is of interest for fundamental understanding and improvement of commercial synthesis of carbon nanotubes (CNTs). As a matter of fact, the method of mechanothermal guarantees the production of MWCNTs suitable for different applications.

  12. Quality and high yield synthesis of Ag nanowires by microwave-assisted hydrothermal method.

    PubMed

    Meléndrez, Manuel F; Medina, Carlos; Solis-Pomar, Francisco; Flores, Paulo; Paulraj, Mani; Pérez-Tijerina, Eduardo

    2015-01-01

    Silver nanowires (Ag-NWs) were obtained using microwave-assisted hydrothermal method (MAH). The main advantage of the method is its high NWs production which is greater than 90%. It is also easy, fast, and highly reproducible process. One of the drawbacks presented so far in the synthesis of nanostructures by polyol path is the high temperature used in the process, which is superior than the boiling point of solvent (ethylene glycol), and also its excessive reaction time. Here, Ag-NWs with diameters of 70 to 110 nm were synthesized in 5 min in large quantities. Results showed that dimensions and shape of nanowires were very susceptible to changes with reaction parameters. The reactor power and reactor fill capacity were important for the synthesis. It was found that the reaction time needs to be decreased because of the NWs which start to deform and break up due to significant increase in the pressure's system. Energy-dispersive X-ray spectroscopy and electron diffraction analysis (SAED) did not show corresponding phases of AgO. Some aspects about synthesis parameters which are related to the percent yield and size of nanowires are also discussed.

  13. Improved yield of high resolution mercuric iodide gamma-ray spectrometers

    SciTech Connect

    Gerrish, V.; van den Berg, L.

    1990-01-01

    Mercuric iodide (HgI{sub 2}) exhibits properties which make it attractive for use as a solid state nuclear radiation detector. The wide bandgap (E{sub g} = 2.1 eV) and low dark current allow room temperature operation, while the high atomic number provides a large gamma-ray cross section. However, poor hole transport has been a major limitation in the routine fabrication of high-resolution spectrometers using this material. This paper presents the results of gamma-ray response and charge transport parameter measurements conducted during the past year at EG G/EM on 96 HgI{sub 2} spectrometers. The gamma-ray response measurements reveal that detector quality is correlated with the starting material used in the crystal growth. In particular, an increased yield of high-resolution spectrometers was obtained from HgI{sub 2} which was synthesized by precipitation from an aqueous solution, as opposed to using material from commercial vendors. Data are also presented which suggest that better spectrometer performance is tied to improved hole transport. Finally, some initial results on a study of detector uniformity reveal spatial variations which may explain why the correlation between hole transport parameters and spectrometer performance is sometimes violated. 6 refs., 3 figs.

  14. 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

  15. 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.

  16. Effect of moderate salinity stress treatment on the stimulation of proline uptake and growth in Escherichia coli CSH4 and its mutants under high salinity.

    PubMed

    Nagata, Shinichi; Wang, Yaoqiang; Zhang, Hongyan; Sasaki, Hideaki; Oshima, Akinobu; Ishida, Akio

    2009-09-01

    Activity of proline uptake in Escherichia coli CSH4 was inhibited in the presence of 1 M NaCl, while it was recovered if the cells were incubated at 30 degrees C for 1 h in a moderate salinity stress (MSS) solution which consists of Davis minimal medium with 5 mM proline and 0.5 M NaCl. Then, an attempt was made to examine whether MSS treatment is also effective on the activity restoration of proline uptake and growth under high salinity for E. coli CSH4 mutants with different combinations of proP, putA, putP, and proU which are related to the transport and metabolization of proline. After MSS treatment, proline uptake was vigorously occurred for the mutants with proline transporter gene proP but not for its deficient ones. For the expression of proline uptake activities of these mutant strains after MSS treatment, PO(4)(3-) in MSS solution is more important than K(+). No growth of strain CSH4 and its mutants without MSS treatment was observed, when cultured in high osmotic medium G (0.8 M NaCl) consisting of 1 mM glycine betaine and Davis minimal medium without potassium phosphate supplemented. After MSS treatment, however, mutant strains lacking proP showed sufficient growth in medium G. Cell growth of proP(+) strains was recognized if MSS treatment was performed in the absence of proline. In conclusion, growth of mutant strains under high-salinity medium G depended on their amount of proline accumulated during MSS treatment, in which K(+) and PO(4)(3-) might play a key role to guarantee their sufficient growth.

  17. Prospects for high-gain, high yield National Ignition Facility targets driven by 2(omega) (green) light

    SciTech Connect

    Suter, L J; Glenzer, S; Haan, S; Hammel, B; Manes, K; Meezan, N; Moody, J; Spaeth, M; Divol, L; Oades, K; Stevenson, M

    2003-12-16

    The National Ignition Facility (NIF), operating at green (2{omega}) light, has the potential to drive ignition targets with significantly more energy than the 1.8 MJ it will produce with its baseline, blue (3{omega}) operations. This results in a greatly increased 'target design space', providing a number of exciting opportunities for fusion research. These include the prospect of ignition experiments with capsules absorbing energies in the vicinity of 1 MJ. This significant increase in capsule absorbed energy over the original designs at {approx}150 kJ could allow high-gain, high yield experiments on NIF. This paper reports the progress made exploring 2{omega} for NIF ignition, including potential 2{omega} laser performance, 2{omega} ignition target designs and 2{omega} Laser Plasma Interaction (LPI) studies.

  18. 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

  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 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

  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

    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.

  1. Facile Growth of High-Yield Gold Nanobipyramids Induced by Chloroplatinic Acid for High Refractive Index Sensing Properties

    PubMed Central

    Fang, Caihong; Zhao, Guili; Xiao, Yanling; Zhao, Jun; Zhang, Zijun; Geng, Baoyou

    2016-01-01

    Au nanobipyramids (NBPs) have attracted great attention because of their unique localized surface plasmon resonance properties. However, the current growth methods always have low yield or suffer tedious process. Developing new ways to direct synthesis of high-yield Au NBPs using common agents is therefore desirable. Here, we employed chloroplatinic acid as the key shape-directing agent for the first time to grow Au NBPs using a modified seed-mediated method at room temperature. H2PtCl6 was added both during the seed preparation and in growth solution. Metallic Pt, reduced from chloroplatinic acid, will deposit on the surface of the seed nanoparticles and the Au nanocrystals and thus plays a critical role for the formation of Au NBPs. Additionally, the reductant, precursor, and surfactant are all cheap and commonly used. Furthermore, the Au NBPs offer narrow size distribution, two sharp tips, and a shared basis. Au NBPs therefore show much higher refractive index sensitivities than that of the Au nanorods. The refractive index sensitivities and lager figure of merit values of Au NBPs exhibit an increase of 63% and 321% respectively compared to the corresponding values of Au nanorod sample. PMID:27841289

  2. Facile Growth of High-Yield Gold Nanobipyramids Induced by Chloroplatinic Acid for High Refractive Index Sensing Properties

    NASA Astrophysics Data System (ADS)

    Fang, Caihong; Zhao, Guili; Xiao, Yanling; Zhao, Jun; Zhang, Zijun; Geng, Baoyou

    2016-11-01

    Au nanobipyramids (NBPs) have attracted great attention because of their unique localized surface plasmon resonance properties. However, the current growth methods always have low yield or suffer tedious process. Developing new ways to direct synthesis of high-yield Au NBPs using common agents is therefore desirable. Here, we employed chloroplatinic acid as the key shape-directing agent for the first time to grow Au NBPs using a modified seed-mediated method at room temperature. H2PtCl6 was added both during the seed preparation and in growth solution. Metallic Pt, reduced from chloroplatinic acid, will deposit on the surface of the seed nanoparticles and the Au nanocrystals and thus plays a critical role for the formation of Au NBPs. Additionally, the reductant, precursor, and surfactant are all cheap and commonly used. Furthermore, the Au NBPs offer narrow size distribution, two sharp tips, and a shared basis. Au NBPs therefore show much higher refractive index sensitivities than that of the Au nanorods. The refractive index sensitivities and lager figure of merit values of Au NBPs exhibit an increase of 63% and 321% respectively compared to the corresponding values of Au nanorod sample.

  3. Error-Prone PCR-Based Mutagenesis Strategy for Rapidly Generating High-Yield Influenza Vaccine Candidates

    PubMed Central

    Ye, Jianqiang; Wen, Feng; Xu, Yifei; Zhao, Nan; Long, Liping; Sun, Hailiang; Yang, Jialiang; Cooley, Jim; Pharr, G. Todd; Webby, Richard; Wan, Xiu-Feng

    2015-01-01

    Vaccination is the primary strategy for the prevention and control of influenza outbreaks. However, the manufacture of influenza vaccine requires a high-yield seed strain, and the conventional methods for generating such strains are time consuming. In this study, we developed a novel method to rapidly generate high-yield candidate vaccine strains by integrating error-prone PCR, site-directed mutagenesis strategies, and reverse genetics. We used this method to generate seed strains for the influenza A(H1N1)pdm09 virus and produced six high-yield candidate strains. We used a mouse model to assess the efficacy of two of the six candidate strains as a vaccine seed virus: both strains provided complete protection in mice against lethal challenge, thus validating our method. Results confirmed that the efficacy of these candidate vaccine seed strains was not affected by the yield-optimization procedure. PMID:25899178

  4. 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.

  5. 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

    2016-12-01

    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.

  6. [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.

  7. 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.

  8. High-throughput and high-yield fabrication of uniaxially-aligned chitosan-based nanofibers by centrifugal electrospinning

    PubMed Central

    Erickson, Ariane E.; Edmondson, Dennis; Chang, Fei-Chien; Wood, Dave; Gong, Alex; Levengood, Sheeny Lan; Zhang, Miqin

    2016-01-01

    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. PMID:26428148

  9. Comparative metabolic flux analysis of an Ashbya gossypii wild type strain and a high riboflavin-producing mutant strain.

    PubMed

    Jeong, Bo-Young; Wittmann, Christoph; Kato, Tatsuya; Park, Enoch Y

    2015-01-01

    In the present study, we analyzed the central metabolic pathway of an Ashbya gossypii wild type strain and a riboflavin over-producing mutant strain developed in a previous study in order to characterize the riboflavin over-production pathway. (13)C-Metabolic flux analysis ((13)C-MFA) was carried out in both strains, and the resulting data were fit to a steady-state flux isotopomer model using OpenFLUX. Flux to pentose-5-phosphate (P5P) via the pentose phosphate pathway (PPP) was 9% higher in the mutant strain compared to the wild type strain. The flux from purine synthesis to riboflavin in the mutant strain was 1.6%, while that of the wild type strain was only 0.1%, a 16-fold difference. In addition, the flux from the cytoplasmic pyruvate pool to the extracellular metabolites, pyruvate, lactate, and alanine, was 2-fold higher in the mutant strain compared to the wild type strain. This result demonstrates that increased guanosine triphosphate (GTP) flux through the PPP and purine synthesis pathway (PSP) increased riboflavin production in the mutant strain. The present study provides the first insight into metabolic flux through the central carbon pathway in A. gossypii and sets the foundation for development of a quantitative and functional model of the A. gossypii metabolic network.

  10. 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.

  11. The tomato homolog of the gene encoding UV-damaged DNA binding protein 1 (DDB1) underlined as the gene that causes the high pigment-1 mutant phenotype.

    PubMed

    Lieberman, Michal; Segev, Orit; Gilboa, Nehama; Lalazar, Avraham; Levin, Ilan

    2004-05-01

    A tomato EST sequence, highly homologous to the human and Arabidopsis thaliana UV-damaged DNA binding protein 1 (DDB1), was mapped to the centromeric region of the tomato chromosome 2. This region was previously shown to harbor the HP-1 gene, encoding the high pigment-1 ( hp-1) and the high pigment-1(w) ( hp-1(w)) mutant phenotypes. Recent results also show that the A. thaliana DDB1 protein interacts both genetically and biochemically with the protein encoded by DEETIOLATED1, a gene carrying three tomato mutations that are in many respects isophenotypic to hp-1: high pigment-2 ( hp-2), high pigment-2(j) ( hp-2(j)) and dark green ( dg). The entire coding region of the DDB1 gene was sequenced in an hp-1 mutant and its near-isogenic normal plant in the cv. Ailsa Craig background, and also in an hp-1(w) mutant and its isogenic normal plant in the GT breeding line background. Sequence analysis revealed a single A(931)-to-T(931) base transversion in the coding sequence of the DDB1 gene in the hp-1 mutant plants. This transversion results in the substitution of the conserved asparagine at position 311 to a tyrosine residue. In the hp-1(w) mutant, on the other hand, a single G(2392)-to-A(2392) transition was observed, resulting in the substitution of the conserved glutamic acid at position 798 to a lysine residue. The single nucleotide polymorphism that differentiates hp-1 mutant and normal plants in the cv. Ailsa Craig background was used to design a pyrosequencing genotyping system. Analysis of a resource F(2) population segregating for the hp-1 mutation revealed a very strong linkage association between the DDB1 locus and the photomorphogenic response of the seedlings, measured as hypocotyl length (25mutant phenotypes.

  12. 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.

  13. An investigation for unexpected high yield of peroxides from isoprene through aqueous phase ozonolysis

    NASA Astrophysics Data System (ADS)

    Wang, H.; Chen, Z.; Hua, W.; Jie, C.

    2007-12-01

    It has recently become evident that isoprene, the atmosphere's most abundant non-methane hydrocarbon, and its oxidation products can considerably result in formation of secondary organic aerosols (SOA) through the acid- catalyzed aqueous phase reaction with hydrogen peroxide. However, the peroxide source in the atmospheric aqueous process is unclear. The present study revealed a potentially important route to the formation of aqueous peroxides, including hydrogen peroxide and hydroxylmethyl hydroperoxide, from the aqueous phase ozonolysis of isoprene. In this study, the atmospheric aqueous phase ozonolysis of isoprene at different pHs and temperatures were studied with the method of laboratory simulation. The major products, including peroxides and carbonyl compounds, were well-characterized, with a measured carbon balance approaching 100%, and the detailed reaction mechanisms were proposed. Most strikingly, peroxides have been found in the aqueous phase ozonolysis of isoprene with unexpected high yields. Considering the huge amount of isoprene in the atmosphere, we suggest that the aqueous phase ozonolysis of isoprene and its first-generation oxidation products may contribute a considerable and even the main source of oxidants to the atmospheric aqueous phase. This means that isoprene and its oxidation products can be transformed into SOA by peroxides provided from their aqueous phase ozonolysis reactions, even if there is no other peroxide source.

  14. 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.

  15. Single PA mutation as a high yield determinant of avian influenza vaccines

    PubMed Central

    Lee, Ilseob; Il Kim, Jin; Park, Sehee; Bae, Joon-Yong; Yoo, Kirim; Yun, Soo-Hyeon; Lee, Joo-Yeon; Kim, Kisoon; Kang, Chun; Park, Man-Seong

    2017-01-01

    Human infection with an avian influenza virus persists. To prepare for a potential outbreak of avian influenza, we constructed a candidate vaccine virus (CVV) containing hemagglutinin (HA) and neuraminidase (NA) genes of a H5N1 virus and evaluated its antigenic stability after serial passaging in embryonated chicken eggs. The passaged CVV harbored the four amino acid mutations (R136K in PB2; E31K in PA; A172T in HA; and R80Q in M2) without changing its antigenicity, compared with the parental CVV. Notably, the passaged CVV exhibited much greater replication property both in eggs and in Madin-Darby canine kidney and Vero cells. Of the four mutations, the PA E31K showed the greatest effect on the replication property of reverse genetically-rescued viruses. In a further luciferase reporter, mini-replicon assay, the PA mutation appeared to affect the replication property by increasing viral polymerase activity. When applied to different avian influenza CVVs (H7N9 and H9N2 subtypes), the PA E31K mutation resulted in the increases of viral replication in the Vero cell again. Taken all together, our results suggest the PA E31K mutation as a single, substantial growth determinant of avian influenza CVVs and for the establishment of a high-yield avian influenza vaccine backbone. PMID:28084423

  16. 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.

  17. Clinical genomics expands the morbid genome of intellectual disability and offers a high diagnostic yield.

    PubMed

    Anazi, S; Maddirevula, S; Faqeih, E; Alsedairy, H; Alzahrani, F; Shamseldin, H E; Patel, N; Hashem, M; Ibrahim, N; Abdulwahab, F; Ewida, N; Alsaif, H S; Al Sharif, H; Alamoudi, W; Kentab, A; Bashiri, F A; Alnaser, M; AlWadei, A H; Alfadhel, M; Eyaid, W; Hashem, A; Al Asmari, A; Saleh, M M; AlSaman, A; Alhasan, K A; Alsughayir, M; Al Shammari, M; Mahmoud, A; Al-Hassnan, Z N; Al-Husain, M; Osama Khalil, R; Abd El Meguid, N; Masri, A; Ali, R; Ben-Omran, T; El Fishway, P; Hashish, A; Ercan Sencicek, A; State, M; Alazami, A M; Salih, M A; Altassan, N; Arold, S T; Abouelhoda, M; Wakil, S M; Monies, D; Shaheen, R; Alkuraya, F S

    2017-04-01

    Intellectual disability (ID) is a measurable phenotypic consequence of genetic and environmental factors. In this study, we prospectively assessed the diagnostic yield of genomic tools (molecular karyotyping, multi-gene panel and exome sequencing) in a cohort of 337 ID subjects as a first-tier test and compared it with a standard clinical evaluation performed in parallel. Standard clinical evaluation suggested a diagnosis in 16% of cases (54/337) but only 70% of these (38/54) were subsequently confirmed. On the other hand, the genomic approach revealed a likely diagnosis in 58% (n=196). These included copy number variants in 14% (n=54, 15% are novel), and point mutations revealed by multi-gene panel and exome sequencing in the remaining 43% (1% were found to have Fragile-X). The identified point mutations were mostly recessive (n=117, 81%), consistent with the high consanguinity of the study cohort, but also X-linked (n=8, 6%) and de novo dominant (n=19, 13%). When applied directly on all cases with negative molecular karyotyping, the diagnostic yield of exome sequencing was 60% (77/129). Exome sequencing also identified likely pathogenic variants in three novel candidate genes (DENND5A, NEMF and DNHD1) each of which harbored independent homozygous mutations in patients with overlapping phenotypes. In addition, exome sequencing revealed de novo and recessive variants in 32 genes (MAMDC2, TUBAL3, CPNE6, KLHL24, USP2, PIP5K1A, UBE4A, TP53TG5, ATOH1, C16ORF90, SLC39A14, TRERF1, RGL1, CDH11, SYDE2, HIRA, FEZF2, PROCA1, PIANP, PLK2, QRFPR, AP3B2, NUDT2, UFC1, BTN3A2, TADA1, ARFGEF3, FAM160B1, ZMYM5, SLC45A1, ARHGAP33 and CAPS2), which we highlight as potential candidates on the basis of several lines of evidence, and one of these genes (SLC39A14) was biallelically inactivated in a potentially treatable form of hypermanganesemia and neurodegeneration. Finally, likely causal variants in previously published candidate genes were identified (ASTN1, HELZ, THOC6, WDR45B, ADRA

  18. Unravelling the mechanisms behind mixed catalysts for the high yield production of single-walled carbon nanotubes.

    PubMed

    Tetali, Sailaja; Zaka, Mujtaba; Schönfelder, Ronny; Bachmatiuk, Alicja; Börrnert, Felix; Ibrahim, Imad; Lin, Jarrn H; Cuniberti, Gianaurelio; Warner, Jamie H; Büchner, Bernd; Rümmeli, Mark H

    2009-12-22

    The use of mixed catalysts for the high-yield production of single-walled carbon nanotubes is well-known. The mechanisms behind the improved yield are poorly understood. In this study, we systematically explore different catalyst combinations from Ni, Co, and Mo for the synthesis of carbon nanotubes via laser evaporation. Our findings reveal that the mixing of catalysts alters the catalyst cluster size distribution, maximizing the clusters' potential to form a hemispherical cap at nucleation and, hence, form a single-walled carbon nanotube. This process significantly improves the single-walled carbon nanotube yields.

  19. RADIATION CHEMISTRY OF HIGH ENERGY CARBON, NEON AND ARGON IONS: INTEGRAL YIELDS FROM FERROUS SULFATE SOLUTIONS

    SciTech Connect

    Christman, E.A.; Appleby, A.; Jayko, M.

    1980-07-01

    Chemical yields of Fe{sup 3+} have been measured from FeSO{sub 4} solutions irradiated in the presence and absence of oxygen with carbon, neon, and argon ions from the Berkeley Bevalac facility. G(Fe{sup 3+}) decreases with increasing beam penetration and with increasing atomic number of the incident ion. The results are compared with current theoretical expectations of the behavior of these particles in an aqueous absorber. The chemical yields are consistently higher than theoretically predicted, by amounts varying from <6.2% (carbon ions) to <13.2% (argon ions). The additional yields are possibly attributable to fragmentation of the primary particle beams.

  20. 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

  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. [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.

  3. Simultaneous achievement of high ethanol yield and titer in Clostridium thermocellum

    SciTech Connect

    Tian, Liang; Papanek, Beth; Olson, Daniel G.; Rydzak, Thomas; Holwerda, Evert K.; Zheng, Tianyong; Zhou, Jilai; Maloney, Marybeth; Jiang, Nannan; Giannone, Richard J.; Hettich, Robert L.; Guss, Adam M.; Lynd, Lee R.

    2016-06-02

    Background Biofuel production from plant cell walls offers the potential for sustainable and economically attractive alternatives to petroleum-based products. Fuels from cellulosic biomass are particularly promising, but would benefit from lower processing costs. Clostridium thermocellum can rapidly solubilize and ferment cellulosic biomass, making it a promising candidate microorganism for consolidated bioprocessing for biofuel production, but increases in product yield and titer are still needed. Results We started with an engineered C. thermocellum strain where the central metabolic pathways to products other than ethanol had been deleted. After two stages of adaptive evolution, an evolved strain was selected with improved yield and titer. On chemically defined medium with crystalline cellulose as substrate, the evolved strain produced 22.4 ± 1.4 g/L ethanol from 60 g/L cellulose. Moreover, the resulting yield was about 0.39 gETOH/gGluc eq, which is 75 % of the maximum theoretical yield. Genome resequencing, proteomics, and biochemical analysis were used to examine differences between the original and evolved strains. Conclusions A two step selection method successfully improved the ethanol yield and the titer. Finaly, this evolved strain has the highest ethanol yield and titer reported to date for C. thermocellum, and is an important step in the development of this microbe for industrial applications.

  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.

  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

    2016-07-29

    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.

  7. Ru(II) complexes of new tridentate ligands: unexpected high yield of sensitized 1O2.

    PubMed

    Liu, Yao; Hammitt, Richard; Lutterman, Daniel A; Joyce, Lauren E; Thummel, Randolph P; Turro, Claudia

    2009-01-05

    Ru(II) complexes possessing new tridentate ligands with extended pi systems, pydppx (3-(pyrid-2'-yl)-11,12-dimethyl-dipyrido[3,2-a:2',3'-c]phenazine) and pydppn (3-(pyrid-2'-yl)-4,5,9,16-tetraaza-dibenzo[a,c]naphthacene), were synthesized and characterized. The investigation of the photophysical properties of the series [Ru(tpy)(n)(L)(2-n)](2+) (L = pydppx, pydppn, n = 0-2) reveals markedly different excited state behavior among the complexes. The Ru(II) complexes possessing the pydppx ligand are similar to the pydppz (3-(pyrid-2'-yl)dipyrido[3,2-a:2',3'-c]phenazine) systems, with a lowest energy metal-to-ligand charge transfer excited state with lifetimes of 1-4 ns. In contrast, the lowest energy excited state in the [Ru(tpy)(n)(pydppn)(2-n)](2+) (n = 0, 1) complexes is a ligand-centered (3)pipi* localized on the pydppn ligand with lifetimes of approximately 20 mus. The [Ru(tpy)(n)(pydppn)(2-n)](2+) (n = 0, 1) complexes are able to generate (1)O(2) with approximately 100% efficiency. Both [Ru(tpy)(pydppn)](2+) and [Ru(pydppn)(2)](2+) bind to DNA, however, the former exhibits a approximately 10-fold greater DNA binding constant than the latter. Efficient DNA photocleavage is observed for [Ru(tpy)(pydppn)](2+), owing to its ability to photosensitize the production of (1)O(2), which can mediate the reactivity. Such high quantum yields of (1)O(2) photosensitization of transition metal complexes may be useful in the design of new systems with long-lived excited states for photodynamic therapy.

  8. 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

  9. 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....

  10. 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.

  11. 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

  12. 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.

  13. Simultaneous achievement of high ethanol yield and titer in Clostridium thermocellum

    DOE PAGES

    Tian, Liang; Papanek, Beth; Olson, Daniel G.; ...

    2016-06-02

    Background Biofuel production from plant cell walls offers the potential for sustainable and economically attractive alternatives to petroleum-based products. Fuels from cellulosic biomass are particularly promising, but would benefit from lower processing costs. Clostridium thermocellum can rapidly solubilize and ferment cellulosic biomass, making it a promising candidate microorganism for consolidated bioprocessing for biofuel production, but increases in product yield and titer are still needed. Results We started with an engineered C. thermocellum strain where the central metabolic pathways to products other than ethanol had been deleted. After two stages of adaptive evolution, an evolved strain was selected with improved yieldmore » and titer. On chemically defined medium with crystalline cellulose as substrate, the evolved strain produced 22.4 ± 1.4 g/L ethanol from 60 g/L cellulose. Moreover, the resulting yield was about 0.39 gETOH/gGluc eq, which is 75 % of the maximum theoretical yield. Genome resequencing, proteomics, and biochemical analysis were used to examine differences between the original and evolved strains. Conclusions A two step selection method successfully improved the ethanol yield and the titer. Finaly, this evolved strain has the highest ethanol yield and titer reported to date for C. thermocellum, and is an important step in the development of this microbe for industrial applications.« less

  14. 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

    2015-06-06

    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.

  15. 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

  16. 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

  17. High-yield boron nitride nanosheets from ‘chemical blowing’: towards practical applications in polymer composites

    NASA Astrophysics Data System (ADS)

    Wang, Xuebin; Pakdel, Amir; Zhi, Chunyi; Watanabe, Kentaro; Sekiguchi, Takashi; Golberg, Dmitri; Bando, Yoshio

    2012-08-01

    An improved ‘chemical blowing’ route presuming atmospheric-pressure pre-treatment and moderate heating rate of designated precursors was developed to synthesize ultra-thin boron nitride (BN) nanosheets with high yield and large lateral dimensions. The yield reached as high as 40 wt% with respect to raw materials (ammonia borane). The strong oxygen-related ultraviolet luminescence together with a blue emission of these BN nanosheets was then documented and analyzed. This implies potential applications in solid-state lighting, ultraviolet lasing and full-color luminescence. Mechanical strength of different polymeric composites with a small fraction of BN nanosheet fillers was dramatically increased by tens of per cent, while high transparency of composite materials was still maintained in the visible optical range. The increased yield and reduced cost of BN nanosheets should promote their wide practical applications in various composites.

  18. 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.

  19. A high-yielding synthesis of allyl glycosides from peracetylated glycosyl donors.

    PubMed

    Khamsi, Jamal; Ashmus, Roger A; Schocker, Nathaniel S; Michael, Katja

    2012-08-01

    β-Configured peracetylated sugars are often used as easily accessible glycosyl donors that are typically activated with common Lewis acids such as boron trifluoride or trimethylsilyltrifluoromethane sulfonate. Often these glycosylations occur with unsatisfactory yields due to incomplete reactions or extensive byproduct formation, primarily as a result of loss of an additional acetyl group generating partially unprotected glycosides. Here we report a simple glycosylation-reacetylation protocol for the generation of predominantly β-configured peracetylated allyl glucoside, -galactoside, -lactoside, and -maltoside with substantially improved reaction yields.

  20. High yield synthesis of 6-(18F)fluoro-L-dopa

    SciTech Connect

    Chirakal, R.; Firnau, G.; Garnett, E.S.

    1986-03-01

    The radiofluorination of L-dopa with (/sup 18/F)F2 was investigated with the purpose of improving the yield of 6-(/sup 18/F)fluoro-L-dopa. When boron trifluoride was added to the reaction mixture in hydrogen fluoride (HF), the yield was increased threefold. Nine millicuries of 6-(/sup 18/F)fluoro-L-dopa were produced from 100 mCi (/sup 18/F)F2 routinely and reliably after 2 hr of preparation. If acetonitrile or water were substituted for HF, little or no 6-fluoro-L-dopa was made.

  1. High-yield hydrogen production by catalytic gasification of coal or biomass

    SciTech Connect

    Hauserman, W.B.

    1992-01-01

    Gasification of coal or wood, catalyzed by soluble metallic cations to maximize reaction rates and hydrogen yields, offers a potential for large-scale, economical hydrogen production with near-commercial technology. With optimum reaction conditions and catalysts, product gas rich in both hydrogen and methane can be used in fuel cells to produce electricity at efficiencies nearly double those of conventional power plant. If plantation silvaculture techniques can produce wood at a raw energy cost competitive with coal, further enhancement of product gas yields may be possible, with zero net contribution of CO{sub 2} to the atmosphere.

  2. A mutant of Mycobacterium tuberculosis lacking the 19-kilodalton lipoprotein Rv3763 is highly attenuated in vivo but retains potent vaccinogenic properties

    PubMed Central

    Henao-Tamayo, Marcela; Junqueira-Kipnis, Ana Paula; Ordway, Diane; Gonzales-Juarrero, Mercedes; Stewart, Graham R.; Young, Douglas B.; Wilkinson, Robert J.; Basaraba, Randall J.; Orme, Ian M.

    2007-01-01

    A mutant of Mycobacterium tuberculosis (Δ19) lacking the 19kDa lipoprotein grows well in culture in vitro but was shown here to be essentially incapable of any significant replication in mice, even in mice lacking the gamma interferon gene. Despite this, mice inoculated with Δ19 were equally protected against an aerosol delivered challenge with M. tuberculosis compared to the conventional BCG vaccine. Cellular responses, including the generation of activated CD4 and CD8 cells secreting gamma interferon, were produced in similar numbers, and lung cells, particularly dendritic macrophages, exhibited high levels of Class-II MHC expression. These data show that despite being highly attenuated, the Δ19 mutant strongly retained vaccinogenic properties. PMID:17804126

  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. Increased Yield of High Purity Recombinant Human Interferon-γ Utilizing Reversed Phase Column Chromatography

    PubMed Central

    Reddy, Praveen K.; Reddy, Srinivasa G.; Narala, Venkata R.; Majee, Sangita S.; Konda, Sudhakar; Gunwar, Sripad; Reddy, Raju C.

    2007-01-01

    Increasing therapeutic applications for recombinant human interferon-γ (rhIFN-γ), an antiviral proinflammatory cytokine, has broadened interest in optimizing methods for its production and purification. We describe a reversed phase chromatography (RPC) procedure using Source-30™ matrix in the purification of rhIFN-γ from Escherichia coli that results in a higher yield than previously reported. The purified rhIFN-γ monomer from the RPC column is refolded in Tris buffer. Optimal refolding occurs at protein concentrations between 50–100 μg/ml. This method yields greater than 90% of the dimer form with a yield of 40 mg g−1 cell mass. Greater than 99% purity is achieved with further purification over a Superdex G-75 column to obtain specific activities of from 2 to 4 × 107 IU/mg protein as determined via cytopathic antiviral assay. The improved yield of rhIFN-γ in a simple chromatographic purification procedure promises to enhance the development and therapeutic application of this biologically potent molecule. PMID:17049266

  5. High-yield, fluoride-free and large-scale synthesis of MIL-101(Cr).

    PubMed

    Zhao, Tian; Jeremias, Felix; Boldog, Ishtvan; Nguyen, Binh; Henninger, Stefan K; Janiak, Christoph

    2015-10-14

    MIL-101(Cr), one of the most important prototypical MOFs, is well investigated and widely used in many scientific fields. With regard to MOF synthesis in general, the addition of a modifier is commonly used to improve the properties of the products. The effect of inorganic (mineral) and organic acid modifiers was thoroughly investigated in the synthesis of MIL-101(Cr) and HNO3 could increase the yield to over 80% of a product with average SBET > 3200 m(2) g(-1) in repeated experiments (from an average of 50% in most published syntheses) in small-scale laboratory synthesis. The large-scale synthesis could use the finding of HNO3 addition and produce MIL-101(Cr) in >100 g quantities with yields near 70% and BET-surface areas near 4000 m(2) g(-1). The addition of acetic acid (CH3COOH) together with seeding could decrease the reaction temperature, the lowest being 160 °C (from typically 220 °C in published procedures), with still relatively good yield and BET surface area of the product. The use of other strong inorganic or weak carboxylic acids as modulators typically caused a decrease in yield and porosity.

  6. 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....

  7. Understanding dry matter and nitrogen accumulation with time-course for high-yielding wheat production in China.

    PubMed

    Meng, Qingfeng; Yue, Shanchao; Chen, Xinping; Cui, Zhenling; Ye, Youliang; Ma, Wenqi; Tong, Yanan; Zhang, Fusuo

    2013-01-01

    Understanding the time-course of dry matter (DM) and nitrogen (N) accumulation in terms of yield-trait relationships is essential to simultaneously increase grain yield and synchronize N demand and N supply. We collected 413 data points from 11 field experiments to address patterns of DM and N accumulation with time in relation to grain yield and management of winter wheat in China. Detailed growth analysis was conducted at the Zadok growth stages (GS) 25 (regreening), GS30 (stem elongation), GS60 (anthesis), and GS100 (maturity) in all experiments, including DM and N accumulation. Grain yield averaged 7.3 Mg ha(-1), ranging from 2.1 to 11.2 Mg ha(-1). The percent N accumulation was consistent prior to DM accumulation, while both DM and N accumulation increased continuously with growing time. Both the highest and fastest DM and N accumulations were observed from stem elongation to the anthesis stage. Significant correlations between grain yield and DM and N accumulation were found at each of the four growth stages, although no positive relationship was observed between grain yield and harvest index or N harvest index. The yield increase from 7-9 Mg ha(-1) to >9 Mg ha(-1) was mainly attributed to increased DM and N accumulation from stem elongation to anthesis. Although applying more N fertilizer increased N accumulation during this stage, DM accumulation was not improved, indicating that N fertilizer management and related agronomic management should be intensified synchronously across the wheat growing season to simultaneously achieve high yields and match N demand and N supply.

  8. High yield single stage conversion of glucose to hydrogen by photofermentation with continuous cultures of Rhodobacter capsulatus JP91.

    PubMed

    Abo-Hashesh, Mona; Desaunay, Nicolas; Hallenbeck, Patrick C

    2013-01-01

    Photofermentative hydrogen (H(2)) production from glucose with the photosynthetic bacterium Rhodobacter capsulatus JP91 (hup(-)) was examined using a photobioreactor operated in continuous mode. Stable and high hydrogen yields on glucose were obtained at three different retention times (HRTs; 24, 48 and 72 h). The H(2) production rates, varying between 0.57 and 0.81 mmol/h, and optical densities (OD(600 nm)) were similar for the different HRTs examined. However, the rate of glucose consumption was influenced by HRT being greater at HRT 24h than HRTs 48 and 72 h. The highest hydrogen yield, 9.0 ± 1.2 mol H(2)/mol glucose, was obtained at 48 h HRT. These results show that single stage photofermentative hydrogen production from glucose using photobioreactors operated in continuous culture mode gives high, nearly stoichiometric yields of hydrogen from glucose, and thus is considerably more promising than either two stage photofermentation or co-culture approaches.

  9. The DENSE AND ERECT PANICLE 1 (DEP1) gene offering the potential in the breeding of high-yielding rice

    PubMed Central

    Xu, Hai; Zhao, Minghui; Zhang, Qun; Xu, Zhengjin; Xu, Quan

    2016-01-01

    The erect panicle model super-rice can rationally transform the solar energy into accumulated organic matter (biomass) and increase grain yield. The phenotype of erect panicle architecture controlled by DENSE AND ERECT PANICLE 1 (DEP1) has been used in rice breeding for nearly a century owing to its high-yield, lodging tolerance with strong stem, reasonable population structure and high nitrogen use efficiency. DEP1 is a G protein γ subunit that is involved in the regulation of erect panicle, number of grains per panicle, nitrogen uptake, and stress-tolerance through the G protein signal pathway. Here we review the development of erect panicle rice varieties, DEP1 alleles and regulatory network, and its physiological and morphological functions. Additionally, the further increasing the yield potential of erect-panicle super-rice, and the development of molecular designing breeding for indica-japonica hybrid rice with the dep1 gene are also prospected. PMID:28163581

  10. Free energy perturbation simulation on transition states and high-activity mutants of human butyrylcholinesterase for (-)-cocaine hydrolysis.

    PubMed

    Yang, Wenchao; Pan, Yongmei; Fang, Lei; Gao, Daquan; Zheng, Fang; Zhan, Chang-Guo

    2010-08-26

    A unified computational approach based on free energy perturbation (FEP) simulations of transition states has been employed to calculate the mutation-caused shifts of the free energy change from the free enzyme to the rate-determining transition state for (-)-cocaine hydrolysis catalyzed by the currently most promising series of mutants of human butyrylcholinesterase (BChE) that contain the A199S/A328W/Y332G mutations. The FEP simulations were followed by Michaelis-Menten kinetics analysis determining the individual k(cat) and K(M) values missing for the A199S/F227A/A328W/Y332G mutant in this series. The calculated mutation-caused shifts of the free energy change from the free enzyme to the rate-determining transition state are in good agreement with the experimental kinetic data, demonstrating that the unified computational approach based on the FEP simulations of the transition states may be valuable for future computational design of new BChE mutants with a further improved catalytic efficiency against (-)-cocaine.

  11. Heterogeneity of yield strain in low-density versus high-density human trabecular bone

    PubMed Central

    Bevill, Grant; Farhamand, Farhad; Keaveny, Tony M.

    2010-01-01

    Understanding the off-axis behavior of trabecular yield strains may lend unique insight into the etiology of fractures since yield strains provide measures of failure independent of elastic behavior. We sought to address anisotropy of trabecular yield strains while accounting for variations in both density and anatomic site and to determine the mechanisms governing this behavior. Cylindrical specimens were cored from vertebral bodies (n=22, BV/TV=0.11±0.02) and femoral necks (n=28, BV/TV=0.22±0.06) with the principal trabecular orientation either aligned along the cylinder axis (on-axis, n=22) or at an oblique angle of 15° or 45° (off-axis, n=28). Each specimen was scanned with micro-CT, mechanically compressed to failure, and analyzed with nonlinear micro-CT-based finite element analysis. Yield strains depended on anatomic site (p=0.03, ANOVA), and the effect of off-axis loading was different for the two sites (p=0.04) – yield strains increased for off-axis loading of the vertebral bone (p=0.04), but were isotropic for the femoral bone (p=0.66). With sites pooled together, yield strains were positively correlated with BV/TV for on-axis loading (R2=58%, p<0.0001), but no such correlation existed for off-axis loading (p=0.79). Analysis of the modulus-BV/TV and strength-BV/TV relationships indicated that, for the femoral bone, the reduction in strength associated with off-axis loading was greater than that for modulus, while the opposite trend occurred for the vertebral bone. The micro-FE analyses indicated that these trends were due to different failure mechanisms for the two types of bone and the different loading modes. Taken together, these results provide unique insight into the failure behavior of human trabecular bone and highlight the need for a multiaxial failure criterion that accounts for anatomic site and bone volume fraction. PMID:19700162

  12. 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.

  13. Effect of acid additives on sugarcane bagasse pyrolysis: Production of high yields of sugars.

    PubMed

    David, Geraldo Ferreira; Perez, Victor Haber; Rodriguez Justo, Oselys; Garcia-Perez, Manuel

    2017-01-01

    The aim of this work was to improve sugarcane bagasse thermochemical conversion to pyrolytic sugars production, particularly to levoglucosan. The experiments were carried out evaluating the effect of acid washing with HNO3 (0.1wt.%) followed by H2SO4 addition (0.1, 0.2 and 0.3wt.%) at pyrolysis temperatures of 350, 400, 450, 500, 550 and 600°C was studied by Py-GC/MS. The experimental results showed that HNO3 washing, followed by H2SO4 concentration of 0.2wt.% at 350°C resulted in an increase in levoglucosan yield between 5 and 7 times the yield obtained when the raw bagasse was processed. Thus, these results are very attractive to improve pyrolytic sugars production in sugarcane bagasse by previously acid treatment to pyrolysis technology.

  14. 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%.

  15. High-Yield Seeded Growth of Monodisperse Pentatwinned Gold Nanoparticles through Thermally Induced Seed Twinning.

    PubMed

    Sánchez-Iglesias, Ana; Winckelmans, Naomi; Altantzis, Thomas; Bals, Sara; Grzelczak, Marek; Liz-Marzán, Luis M

    2017-01-11

    We show that thermal treatment of small Au seeds results in extensive twinning and a subsequent drastic improvement in the yield (>85%) of formation of pentatwinned nanoparticles (NPs), with preselected morphology (nanorods, bipyramids, and decahedra) and aspect ratio. The "quality" of the seeds thus defines the yield of the obtained NPs, which in the case of nanorods avoids the need for additives such as Ag(+) ions. This modified seeded growth method also improves reproducibility, as the seeds can be stored for extended periods of time without compromising the quality of the final NPs. Additionally, minor modification of the seeds with Pd allows their localization within the final particles, which opens new avenues toward mechanistic studies. Together, these results represent a paradigm shift in anisotropic gold NP synthesis.

  16. Production of high hydroxytyrosol yields via tyrosol conversion by Pseudomonas aeruginosa immobilized resting cells.

    PubMed

    Bouallagui, Zouhaier; Sayadi, Sami

    2006-12-27

    An immobilized whole cell system was successfully performed to produce the most powerful antioxidant, hydroxytyrosol. Bioconversion of tyrosol into hydroxytyrosol was achieved via the immobilization of Pseudomonas aeruginosa resting cells in calcium alginate beads. Immobilization was advantageous as it allows immobilized cells to tolerate a greater tyrosol concentration than free cells. The bioconversion yield reached 86% in the presence of 5 g L-1 of tyrosol when cells immobilized in alginate beads were carried out in single batches. Evaluation of kinetic parameters showed the maintenance of the same catalytic efficiency expressed as Kcat/Km for both free and immobilized cells. The use of immobilized cells in repeated batches demonstrated a notable activity stabilization since the biocatalyst reusability was extended for at least four batches with a molar yield greater than 85%.

  17. 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...

  18. 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...

  19. ‘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...

  20. 'Caro-Tex 312’ – An F1 Hybrid, High Yielding, Multiple Disease Resistant, Orange Habanero Pepper Cultivar

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Texas A&M University and the USDA-ARS U.S. Vegetable Laboratory in Charleston, SC, have developed a new, F1 hybrid Habanero pepper cultivar. ‘Caro-Tex 312’ produces a large, orange-fruited Habanero pepper with typical shape and high pungency. It also possesses unique yield, early maturity and dise...

  1. 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...

  2. Highland Russet: A Full Season, Processing Variety with High Yields of Uniform U.S. No. 1 Tubers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Highland Russet is a late-season potato variety with light russet skin notable for its high yield of uniform U.S. No. 1 tubers, and good processing and culinary qualities. It resulted from a 1990 cross between Ranger Russet and Russet Legend and has been evaluated for over 15 years in public and ind...

  3. Organocatalytic sequential α-amination/Corey-Chaykovsky reaction of aldehydes: a high yield synthesis of 4-hydroxypyrazolidine derivatives.

    PubMed

    Kumar, B Senthil; Venkataramasubramanian, V; Sudalai, Arumugam

    2012-05-18

    A tandem reaction of in situ generated α-amino aldehydes with dimethyloxosulfonium methylide under Corey-Chaykovsky reaction conditions proceeds efficiently to give 4-hydroxypyrazolidine derivatives in high yields with excellent enantio- and diastereoselectivities. This organocatalytic sequential method provides for the efficient synthesis of anti-1,2-aminoalcohols, structural subunits present in several bioactive molecules as well.

  4. High yield secretion of recombinant proteins from the microalga Chlamydomonas reinhardtii.

    PubMed

    Ramos-Martinez, E M; Fimognari, L; Sakuragi, Y

    2017-02-16

    Microalga-based biomanufacturing of recombinant proteins is attracting growing attention due to its advantages in safety, metabolic diversity, scalability, and sustainability. Secretion of recombinant proteins can accelerate the use of microalgal platforms by allowing post-translational modifications and easy recovery of products from the culture media. However, currently, the yields of secreted recombinant proteins are low, which hampers the commercial application of this strategy. This study aimed at expanding the genetic tools for enhancing secretion of recombinant proteins in Chlamydomonas reinhardtii, a widely used green microalga as a model organism and a potential industrial biotechnology platform. We demonstrated that the putative signal sequence from C. reinhardtii gametolysin can assist the secretion of the yellow fluorescent protein Venus into the culture media. In order to increase the secretion yields, Venus was C-terminally fused with synthetic glycomodules comprised of tandem serine (Ser) and proline (Pro) repeats of 10 and 20 units [hereafter (SP)n, wherein n=10 or 20]. The yields of the (SP)n-fused Venus were higher than Venus without the glycomodule by up to 12 folds, with the maximum yield of 15 mg L(-1) . Moreover, the presence of the glycomodules confererred an enhanced proteolytic protein stability. The Venus-(SP)n proteins were shown to be glycosylated, and a treatment of the cells with Brefeldin A led to a suggestion that glycosylation of the (SP)n glycomodules starts in the endoplasmic reticulum (ER). Taken together, the results demonstrate the utility of the gametolysin signal sequence and (SP)n glycomodule to promote a more efficient biomanufacturing of microalgae-based recombinant proteins. This article is protected by copyright. All rights reserved.

  5. Investigation of Ozone Yield of Air Fed Ozonizer by High Pressure Homogeneous Dielectric Barrier Discharge

    DTIC Science & Technology

    2013-07-01

    homogeneous dielectric barrier discharge ( DBD ) in dry air by using a simple DBD device. So far, we have tried to apply the homogeneous DBD to an...specific input energy region. In this work, we investigated the effect of gas pressure (from 0.1 MPa to 0.2 MPa) on the ozone yield by homogeneous DBD . The...homogeneous DBD decreased with increasing the gas pressure. 1. Introduction The dielectric barrier discharge ( DBD ) is composed of many filamentary micro

  6. 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.

  7. A new copper containing MALDI matrix that yields high abundances of [peptide + Cu]+ ions.

    PubMed

    Wu, Zhaoxiang; Fernandez-Lima, Francisco A; Perez, Lisa M; Russell, David H

    2009-07-01

    The dinuclear copper complex (alpha-cyano-4-hydroxycinnamic acid (CHCA) copper salt (CHCA)(4)Cu(2)), synthesized by reacting CHCA with copper oxide (CuO), yields increased abundances of [M + xCu - (x-1)H](+) (x = 1-6) ions when used as a matrix for matrix-assisted laser desorption ionization (355 nm Nd:YAG laser). The yield of [M + xCu - (x-1)H](+) (x = 1 to approximately 6) ion is much greater than that obtained by mixing peptides with copper salts or directly depositing peptides onto oxidized copper surfaces. The increased ion yields for [M + xCu - (x-1)H](+) facilitate studies of biologically important copper binding peptides. For example, using this matrix we have investigated site-specific copper binding of several peptides using fragmentation chemistry of [M + Cu](+) and [M + 2Cu - H](+) ions. The fragmentation studies reveal interesting insight on Cu binding preferences for basic amino acids. Most notable is the fact that the binding of a single Cu(+) ion and two Cu(+) ions are quite different, and these differences are explained in terms of intramolecular interactions of the peptide-Cu ionic complex.

  8. 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.

  9. Enhanced lipid production in thermo-tolerant mutants of Chlorella pyrenoidosa NCIM 2738.

    PubMed

    Sachdeva, Neha; Gupta, Ravi Prakash; Mathur, Anshu Shankar; Tuli, Deepak Kumar

    2016-12-01

    The present study aimed to develop thermo-tolerant mutants of Chlorella pyrenoidosa NCIM 2738 for high lipids production. For this, ethyl methane sulfonate was used, which generated two effective thermo-tolerant mutants, M18 and M24 of Chlorella pyrenoidosa NCIM 2738, capable of surviving at temperature up to 47°C and showing improved lipid and biomass yields. They showed 59.62% and 50.75% increase, respectively in lipid content compared to wild type at 30°C, which could not grow at temperature above 35°C. The novelty of this study lied in incorporation of PAM Flurometry with mutagenesis to generate thermo-tolerant mutants of C. pyrenoidosa and investigating the reasons for increased yields of mutants at cellular and photosynthetic levels with the aim to use them for commercial biodiesel production.

  10. 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.

  11. Failure And Yield Analysis Techniques For Readout Devices Tested In A High Throughput Automated Wafer Probing Environment

    NASA Astrophysics Data System (ADS)

    Jolivet, Noel D.; Holoien, Lee D.

    1990-01-01

    Failure and yield analysis capabilities were developed for focal plane array (FPA) readout devices probe tested at wafer level instead of merely performing production rate testing and cataloging data. Innovative test strategies have been combined with software techniques to provide tools which accomplish these analyses while maintaining high throughput in test. This approach has been beneficial and valuable in saving test time when searching for hardware faults, investigating design susceptibilities, revealing foundry process variations from wafer to wafer and/or lot to lot, and creating a yield model for the parts tested. Testing of readout devices has historically been viewed as a major obstacle in high volume production of reliable components for focal plane systems. Thorough testing in a high throughput automated wafer probe environment may be achieved, but often at the expense of real-time analysis of failures and yield factors. Production testing has been established with these goals in mind rather than as an afterthought. This enables one to identify failure mechanisms as they occur in order to reduce yield loss and unnecessary test time. In addition to performing requisite data base management, routines have been created to re-sort data and reevaluate yield through varying performance parameter limits, to track and map failure mechanisms as they are encountered, to examine acquired data as a function of limits, and to provide yield information for feedback response to foundry processing. Ideas for aiding operators in recognizing and diagnosing possible test set hardware problems (as opposed to on-chip failures) have also been implemented.

  12. 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.

  13. 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.

  14. 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

  15. Demonstration of Radiation Pulse Shaping with Nested-Tungsten-Wire-Array Z Pinches for High-Yield Inertial Confinement Fusion

    SciTech Connect

    Cuneo, M.E.; Vesey, R.A.; Sinars, D.B.; Waisman, E.M.; Lemke, R.W.; Bliss, D.E.; Stygar, W.A.; Porter, J.L.; Mazarakis, M.G.; Chandler, G.A.; Mehlhorn, T.A.; Chittenden, J.P.; Lebedev, S.V.; Schroen, D.G.

    2005-10-28

    Nested wire-array Z pinches are shown to generate soft x-ray radiation pulse shapes required for three-shock isentropic compression and hot-spot ignition of high-yield inertial confinement fusion capsules. We demonstrate a reproducible and tunable foot pulse (first shock) produced by interaction of the outer and inner arrays. A first-step pulse (second shock) is produced by inner array collision with a central CH{sub 2} foam target. Stagnation of the inner array at the axis produces the third shock. Capsules optimized for several of these shapes produce 290-900 MJ fusion yields in 1D simulations.

  16. 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.

  17. High-yield synthesis of boron nitride nanoribbons via longitudinal splitting of boron nitride nanotubes by potassium vapor.

    PubMed

    Sinitskii, Alexander; Erickson, Kristopher J; Lu, Wei; Gibb, Ashley L; Zhi, Chunyi; Bando, Yoshio; Golberg, Dmitri; Zettl, Alex; Tour, James M

    2014-10-28

    Boron nitride nanoribbons (BNNRs) are theorized to have interesting electronic and magnetic properties, but their high-yield synthesis remains challenging. Here we demonstrate that potassium-induced splitting of BN nanotubes (BNNTs) is an effective high-yield method to obtain bulk quantities of high-quality BNNRs if a proper precursor material is chosen. The resulting BNNRs are crystalline; many of them have a high aspect ratio and straight parallel edges. We have observed numerous few-layer and monolayer BNNRs; the multilayered ribbons predominantly have an AA' stacking. We present a detailed microscopy study of BNNRs that provides important insights into the mechanism of the formation of BNNRs from BNNTs. We also demonstrate that the BNNTs prepared by different synthetic approaches could exhibit dramatically different reactivities in the potassium splitting reaction, which highlights the need for future comparison studies of BN nanomaterials prepared using different methods to better understand their preparation-dependent physical and chemical properties.

  18. 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 ...

  19. Robust root growth in altered hydrotropic response1 (ahr1) mutant of Arabidopsis is maintained by high rate of cell production at low water potential gradient.

    PubMed

    Salazar-Blas, Amed; Noriega-Calixto, Laura; Campos, María E; Eapen, Delfeena; Cruz-Vázquez, Tania; Castillo-Olamendi, Luis; Sepulveda-Jiménez, Gabriela; Porta, Helena; Dubrovsky, Joseph G; Cassab, Gladys I

    2017-01-01

    Hydrotropism is the directional root growth response determined by water stimulus. In a water potential gradient system (WPGS) the roots of the Arabidopsis wild type have a diminished root growth compared to normal medium (NM). In contrast, the altered hydrotropic response1 (ahr1) mutant roots maintain their robust growth in the same WPGS. The aims of this work were to ascertain how ahr1 roots could sustain growth in the WPGS, with a special focus on the integration of cellular processes involved in the signaling that determines root growth during abiotic stress and their relation to hydrotropism. Cellular analysis of the root apical meristem of ahr1 mutant contrary to the wild type showed an absence of changes in the meristem length, the elongation zone length, the length of fully elongated cells, and the cell cycle duration. The robust and steady root growth of ahr1 seedlings in the WPGS is explained by the mutant capacity to maintain cell production and cell elongation at the same level as in the NM. Analysis of auxin response at a transcriptional level showed that roots of the ahr1 mutant had a lower auxin response when grown in the WPGS, compared to wild type, indicating that auxin signaling participates in attenuation of root growth under water stress conditions. Also, wild type plants exhibited a high increase in proline content while ahr1 mutants showed minimum changes in the Normal Medium→Water Stress Medium (NM→WSM), a lower water potential gradient system than the WPGS. Accordingly, in this condition, gene expression of Δ1-6 Pyrroline-5-Carboxylate Synthetase1 (P5CS1) involved in proline synthesis strongly increased in wild type but not in ahr1 seedlings. The ahr1 phenotype shows unique features since the mutant root cells continue to proliferate and grow in the presence of a progressively negative water potential gradient at a level comparable to wild type growing in the NM. As such, it represents an exceptional resource for understanding

  20. 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

    2011-04-07

    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.

  1. High-yield thin GaN LED using metal bonding and laser lift-off technology

    NASA Astrophysics Data System (ADS)

    Horng, Ray-Hua; Chen, Ching-Ho; Kao, Wei-Cheng; Wuu, Dong-Sing

    2012-10-01

    Gold-indium metal bonding method was used in this study to increase the product yield of vertical light emitting diodes (LEDs) during laser lift-off (LLO) process. The vertical GaN LED transferred onto Si substrate presented good electrical and optical properties due to the existence of high reflective mirror and texture surface. The chip size and dominant wavelength for vertical type LED are 40×40 mil2 and 450 nm. The optimal conditions of temperature and pressure for 2-inch wafer bonding are set of 200oC and 100 kg/inch2, respectively. The products yield of light output power, forward voltage and leakage current are 96 %, 96.4% and 61.2%, respectively. After aging test, the characteristics decay of light output power, forward voltage and leakage current are less than 4%. Summarization of optical and electrical properties, the total yield of these LEDs products is about 60 %.

  2. Repression of Invasion Genes and Decreased Invasion in a High-Level Fluoroquinolone-Resistant Salmonella Typhimurium Mutant

    PubMed Central

    Fàbrega, Anna; du Merle, Laurence; Le Bouguénec, Chantal; Jiménez de Anta, M. Teresa; Vila, Jordi

    2009-01-01

    Background Nalidixic acid resistance among Salmonella Typhimurium clinical isolates has steadily increased, whereas the level of ciprofloxacin resistance remains low. The main objective of this study was to characterize the fluoroquinolone resistance mechanisms acquired in a S. Typhimurium mutant selected with ciprofloxacin from a susceptible isolate and to investigate its invasion ability. Methodology/Principal Findings Three different amino acid substitutions were detected in the quinolone target proteins of the resistant mutant (MIC of ciprofloxacin, 64 µg/ml): D87G and G81C in GyrA, and a novel mutation, E470K, in ParE. A protein analysis revealed an increased expression of AcrAB/TolC and decreased expression of OmpC. Sequencing of the marRAB, soxRS, ramR and acrR operons did not show any mutation and neither did their expression levels in a microarray analysis. A decreased percentage of invasion ability was detected when compared with the susceptible clinical isolate in a gentamicin protection assay. The microarray results revealed a decreased expression of genes which play a role during the invasion process, such as hilA, invF and the flhDC operon. Of note was the impaired growth detected in the resistant strain. A strain with a reverted phenotype (mainly concerning the resistance phenotype) was obtained from the resistant mutant. Conclusions/Significance In conclusion, a possible link between fluoroquinolone resistance and decreased cell invasion ability may exist explaining the low prevalence of fluoroquinolone-resistant S. Typhimurium clinical isolates. The impaired growth may appear as a consequence of fluoroquinolone resistance acquisition and down-regulate the expression of the invasion genes. PMID:19946377

  3. 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

  4. Vineyard yield estimation based on the analysis of high resolution images obtained with artificial illumination at night.

    PubMed

    Font, Davinia; Tresanchez, Marcel; Martínez, Dani; Moreno, Javier; Clotet, Eduard; Palacín, Jordi

    2015-04-09

    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.

  5. 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.

  6. Native Promoter Strategy for High-Yielding Synthesis and Engineering of Fungal Secondary Metabolites

    PubMed Central

    2015-01-01

    Strategies are needed for the robust production of cryptic, silenced, or engineered secondary metabolites in fungi. The filamentous fungus Fusarium heterosporum natively synthesizes the polyketide equisetin at >2 g L–1 in a controllable manner. We hypothesized that this production level was achieved by regulatory elements in the equisetin pathway, leading to the prediction that the same regulatory elements would be useful in producing other secondary metabolites. This was tested by using the native eqxS promoter and eqxR regulator in F. heterosporum, synthesizing heterologous natural products in yields of ∼1 g L–1. As proof of concept for the practical application, we resurrected an extinct pathway from an endophytic fungus with an initial yield of >800 mg L–1, leading to the practical synthesis of a selective antituberculosis agent. Finally, the method enabled new insights into the function of polyketide synthases in filamentous fungi. These results demonstrate a strategy for optimally employing native regulators for the robust synthesis of secondary metabolites. PMID:25226362

  7. Native promoter strategy for high-yielding synthesis and engineering of fungal secondary metabolites.

    PubMed

    Kakule, Thomas B; Jadulco, Raquel C; Koch, Michael; Janso, Jeffrey E; Barrows, Louis R; Schmidt, Eric W

    2015-05-15

    Strategies are needed for the robust production of cryptic, silenced, or engineered secondary metabolites in fungi. The filamentous fungus Fusarium heterosporum natively synthesizes the polyketide equisetin at >2 g L(-1) in a controllable manner. We hypothesized that this production level was achieved by regulatory elements in the equisetin pathway, leading to the prediction that the same regulatory elements would be useful in producing other secondary metabolites. This was tested by using the native eqxS promoter and eqxR regulator in F. heterosporum, synthesizing heterologous natural products in yields of ∼1 g L(-1). As proof of concept for the practical application, we resurrected an extinct pathway from an endophytic fungus with an initial yield of >800 mg L(-1), leading to the practical synthesis of a selective antituberculosis agent. Finally, the method enabled new insights into the function of polyketide synthases in filamentous fungi. These results demonstrate a strategy for optimally employing native regulators for the robust synthesis of secondary metabolites.

  8. 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.

  9. Spectroscopy of colloidal semiconductor core/shell nanoplatelets with high quantum yield.

    PubMed

    Tessier, M D; Mahler, B; Nadal, B; Heuclin, H; Pedetti, S; Dubertret, B

    2013-07-10

    Free standing two-dimensional materials appear as a novel class of structures. Recently, the first colloidal two-dimensional heterostructures have been synthesized. These core/shell nanoplatelets are the first step toward colloidal quantum wells. Here, we study in detail the spectroscopic properties of this novel generation of colloidal nanoparticles. We show that core/shell CdSe/CdZnS nanoplatelets with 80% quantum yield can be obtained. The emission time trace of single core/shell nanoplatelets exhibits reduced blinking compared to core nanoplatelets with a two level emission time trace. At cryogenic temperatures, these nanoplatelets have a quantum yield close to 100% and a stable emission time trace. A solution of core/shell nanoplatelets has emission spectra with a full width half-maximum close to 20 nm, a value much lower than corresponding spherical or rod-shaped heterostructures. Using single particle spectroscopy, we show that the broadening of the emission spectra upon the shell deposition is not due to dispersity between particles but is related to an intrinsic increased exciton-phonon coupling in the shell. We also demonstrate that optical spectroscopy is a relevant tool to investigate the presence of traps induced by shell deposition. The spectroscopic properties of the core/shell nanoplatelets presented here strongly suggest that this new generation of objects will be an interesting alternative to spherical or rod-shaped nanocrystals.

  10. High temperature pre-digestion of corn stover biomass for improved product yields

    DOE PAGES

    Brunecky, Roman; Hobdey, Sarah E.; Taylor, Larry E.; ...

    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

  11. A high yield multi-method extraction protocol for protein quantification in activated sludge.

    PubMed

    Monique, Ras; Elisabeth, Girbal-Neuhauser; Etienne, Paul; Dominique, Lefebvre

    2008-11-01

    A multi-method extraction protocol based on mechanical, ionic and hydrophobic methods was investigated on two types of activated sludge samples. Extraction methods were chosen with regards to optimal protein yield without cell disruption. Sonication, EDTA and Tween extraction methods were selected and combined. The total amount of protein released by the multi-method protocol sums up to 191 and 264 mg equiv. BSA/g VSS for the two different sludge samples. Protocol repetition on the same sample showed that protein yield after each successive protocol fitted an exponential curve model. The total amount of extractable proteins was evaluated by model predictions, 423 and 516 mg equiv. BSA/g VSS for the two sludge samples. The multi-method extraction protocol appears relevant for harvesting a representative quantity of proteins from the original sample (45-49%), moreover the multi-method criterion of the protocol also offers a heterogeneous pool of proteins. Thus, further qualitative studies may not be biased by the extraction protocol.

  12. A High-Throughput Screening Method for Small-Molecule Inhibitors of the Aberrant Mutant SOD1 and Dynein Complex Interaction

    PubMed Central

    Tang, Xiaohu; Seyb, Kathleen I.; Huang, Mickey; Schuman, Eli R.; Shi, Ping; Zhu, Haining; Glicksman, Marcie A.

    2013-01-01

    Aberrant protein-protein interactions are attractive drug targets in a variety of neurodegenerative diseases due to the common pathology of accumulation of protein aggregates. In amyotrophic lateral sclerosis, mutations in SOD1 cause the formation of aggregates and inclusions that may sequester other proteins and disrupt cellular processes. It has been demonstrated that mutant SOD1, but not wild-type SOD1, interacts with the axonal transport motor dynein and that this interaction contributes to motor neuron cell death, suggesting that disrupting this interaction may be a potential therapeutic target. However, it can be challenging to configure a high-throughput screening (HTS)–compatible assay to detect inhibitors of a protein-protein interaction. Here we describe the development and challenges of an HTS for small-molecule inhibitors of the mutant SOD1-dynein interaction. We demonstrate that the interaction can be formed by coexpressing the A4V mutant SOD1 and dynein intermediate complex in cells and that this interaction can be disrupted by compounds added to the cell lysates. Finally, we show that some of the compounds identified from a pilot screen to inhibit the protein-protein interaction with this method specifically disrupt the interaction between the dynein complex and mtSOD1 but not the dynein complex itself when applied to live cells. PMID:22140121

  13. High-Throughput Genotyping of Green Algal Mutants Reveals Random Distribution of Mutagenic Insertion Sites and Endonucleolytic Cleavage of Transforming DNA.

    PubMed

    Zhang, Ru; Patena, Weronika; Armbruster, Ute; Gang, Spencer S; Blum, Sean R; Jonikas, Martin C

    2014-04-01

    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.

  14. Methylosinus trichosporium OB3b Mutants Having Constitutive Expression of Soluble Methane Monooxygenase in the Presence of High Levels of Copper

    PubMed Central

    Phelps, Patricia A.; Agarwal, Sandeep K.; Speitel, Gerald E.; Georgiou, George

    1992-01-01

    The methanotrophic bacterium Methylosinus trichosporium OB3b is unusually active in degrading recalcitrant haloalkanes such as trichloroethylene (TCE). The first and rate-limiting step in the degradation of TCE is catalyzed by a soluble methane monooxygenase (sMMO). This enzyme is not expressed when the cells are grown in the presence of copper at concentrations typically found in polluted groundwater. Under these conditions, M. trichosporium OB3b expresses a particulate form of the enzyme (pMMO), which has a narrow substrate specificity and does not degrade TCE at any significant rate. We have isolated M. trichosporium OB3b mutants that are deficient in pMMO and express sMMO constitutively in the presence of elevated concentrations of copper. One mutant (PP358) exhibited a TCE degradation rate which was almost twice as high as that of the wild-type strain grown under optimal conditions (without copper). All of the mutants lost the ability to express pMMO activity and to form stacked intracellular membranes characteristic of wild-type cells expressing pMMO. Images PMID:16348810

  15. A Brucella melitensis high-temperature-requirement A (htrA) deletion mutant is attenuated in goats and protects against abortion.

    PubMed

    Phillips, R W; Elzer, P H; Robertson, G T; Hagius, S D; Walker, J V; Fatemi, M B; Enright, F M; Roop, R M

    1997-01-01

    It has been previously demonstrated that a Brucella melitensis high-temperature-requirement A (htrA) deletion mutant is more susceptible to oxidative killing in vitro than the parental strain and is attenuated in mice. To evaluate the contribution of the B melitensis HtrA protease to virulence in ruminants, the capacity of the B melitensis htrA mutant RWP5 to produce abortion in goats was compared to that of the virulent parental strain 16M. Experimental infection with strain 16M caused abortion in eight of 12 pregnant nannies, while none of the 12 nannies inoculated with RWP5 aborted. Furthermore, intramuscular injection of fetuses in utero with RWP5 led to colonisation of the fetus with subsequent colonisation of the nanny, but no abortion was observed. Nannies vaccinated with RWP5 showed complete protection against abortion when challenged with 16M during the third trimester of pregnancy. However, these animals were not protected from colonisation by 16M. The results presented here clearly indicate that the B melitensis htrA gene product contributes to pathogenesis in goats, but the utility of B melitensis htrA mutants as vaccines in this host appears to be limited.

  16. 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

  17. High-resolution structures of Neotermes koshunensis β-glucosidase mutants provide insights into the catalytic mechanism and the synthesis of glucoconjugates.

    PubMed

    Jeng, Wen-Yih; Wang, Nai-Chen; Lin, Cheng-Tse; Chang, Wei-Jung; Liu, Chia-I; Wang, Andrew H-J

    2012-07-01

    NkBgl, a β-glucosidase from Neotermes koshunensis, is a β-retaining glycosyl hydrolase family 1 enzyme that cleaves β-glucosidic linkages in disaccharide or glucose-substituted molecules. β-Glucosidases have been widely used in several applications. For example, mutagenesis of the attacking nucleophile in β-glucosidase has been conducted to convert it into a glycosynthase for the synthesis of oligosaccharides. Here, several high-resolution structures of wild-type or mutated NkBgl in complex with different ligand molecules are reported. In the wild-type NkBgl structures it was found that glucose-like glucosidase inhibitors bind to the glycone-binding pocket, allowing the buffer molecule HEPES to remain in the aglycone-binding pocket. In the crystal structures of NkBgl E193A, E193S and E193D mutants Glu193 not only acts as the catalytic acid/base but also plays an important role in controlling substrate entry and product release. Furthermore, in crystal structures of the NkBgl E193D mutant it was found that new glucoconjugates were generated by the conjugation of glucose (hydrolyzed product) and HEPES/EPPS/opipramol (buffer components). Based on the wild-type and E193D-mutant structures of NkBgl, the glucosidic bond of cellobiose or salicin was hydrolyzed and a new bond was subsequently formed between glucose and HEPES/EPPS/opipramol to generate new glucopyranosidic products through the transglycosylation reaction in the NkBgl E193D mutant. This finding highlights an innovative way to further improve β-glucosidases for the enzymatic synthesis of oligosaccharides.

  18. Heat shock transcriptional responses in an MC-Producing Cyanobacterium (Planktothrix agardhii) and its MC-deficient mutant under high light conditions.

    PubMed

    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.

  19. Transcriptome Analysis Suggests That Starch Synthesis May Proceed via Multiple Metabolic Routes in High Yielding Potato Cultivars

    PubMed Central

    Kaminski, Kacper Piotr; Petersen, Annabeth Høgh; Sønderkær, Mads; Pedersen, Lars Haastrup; Pedersen, Henrik; Feder, Christian; Nielsen, Kåre L.

    2012-01-01

    Background Glucose-6-phosphate is imported into the amyloplast of potato tubers and thought to constitute the precursor for starch synthesis in potato tubers. However, recently it was shown that glucose-1-phosphate can also be imported into the amyloplast and incorporated into starch via an ATP independent mechanism under special conditions. Nonetheless, glucose-6-phosphate is believed to be the quantitatively important precursor for starch synthesis in potato. Principal Finding Potato tubers of the high yielding cv Kuras had low gene expression of plastidial phophoglucomutase (PGM) and normal levels of transcripts for other enzymes involved in starch metabolism in comparison with medium and low yielding cultivars as determined by DeepSAGE transcriptome profiling. The decrease in PGM activity in Kuras was confirmed by measuring the enzyme activity from potato tuber extracts. Contrary to expectations, this combination lead to a higher level of intracellular glucose-1-phosphate (G1P) in Kuras suggesting that G1P is directly imported into plastids and can be quantitatively important for starch synthesis under normal conditions in high yielding cultivars. Significance This could open entirely new possibilities for metabolic engineering of the starch metabolism in potato via the so far uncharacterized G1P transporter. The perspectives are to increase yield and space efficiency of this important crop. In the light of the increasing demands imposed on agriculture to support a growing global population this presents an exciting new possibility. PMID:23284672

  20. Comparison of screening methods for high-throughput determination of oil yields in micro-algal biofuel strains.

    PubMed

    Slocombe, Stephen P; Zhang, Qianyi; Black, Kenneth D; Day, John G; Stanley, Michele S

    2013-08-01

    The phenotypic and phylogenetic diversity of micro-algae capable of accumulating triacylglycerols provides a challenge for the accurate determination of biotechnological potential. High-yielding strains are needed to improve economic viability and their compositional information is required for optimizing biodiesel properties. To facilitate a high-throughput screening programme, a very rapid direct-derivatization procedure capable of extracting lyophilized material for GC analysis was compared with a scaled-down Folch-based method. This was carried out on ten micro-algal strains from 6 phyla where the more rapid direct-derivatization approach was found to provide a more reliable measure of yield. The modified Folch-based procedure was found to substantially underestimate oil yield in one Chlorella species (P < 0.01). In terms of fatty acid composition however, the Folch procedure proved to be slightly better in recovering polyunsaturated fatty acids, in six out of the ten strains. Therefore, direct-derivatization is recommended for rapid determination of yields in screening approaches but can provide slightly less compositional accuracy than solvent-based extraction methods.

  1. High-yield secretion of recombinant proteins expressed in tobacco cell culture with a designer glycopeptide tag: Process development.

    PubMed

    Zhang, Ningning; Gonzalez, Maria; Savary, Brett; Xu, Jianfeng

    2016-03-01

    Low-yield protein production remains the most significant economic hurdle with plant cell culture technology. Fusions of recombinant proteins with hydroxyproline-O-glycosylated designer glycopeptide tags have consistently boosted secreted protein yields. This prompted us to study the process development of this technology aiming to achieve productivity levels necessary for commercial viability. We used a tobacco BY-2 cell culture expressing EGFP as fusion with a glycopeptide tag comprised of 32 repeat of "Ser-Pro" dipeptide, or (SP)32 , to study cell growth and protein secretion, culture scale-up, and establishment of perfusion cultures for continuous production. The BY-2 cells accumulated low levels of cell biomass (~7.5 g DW/L) in Schenk & Hildebrandt medium, but secreted high yields of (SP)32 -tagged EGFP (125 mg/L). Protein productivity of the cell culture has been stable for 6.0 years. The BY-2 cells cultured in a 5-L bioreactor similarly produced high secreted protein yield at 131 mg/L. Successful operation of a cell perfusion culture for 30 days was achieved under the perfusion rate of 0.25 and 0.5 day(-1) , generating a protein volumetric productivity of 17.6 and 28.9 mg/day/L, respectively. This research demonstrates the great potential of the designer glycopeptide technology for use in commercial production of valuable proteins with plant cell cultures.

  2. 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.

  3. Optimizing hill seeding density for high-yielding hybrid rice in a single rice cropping system in South China.

    PubMed

    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.

  4. 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

  5. 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...

  6. Enhanced conversion of sucrose to isomaltulose by a mutant of Erwinia rhapontici.

    PubMed

    Ahn, Seung-Joon; Yoo, Ji-Hyun; Lee, Hyeon-Cheol; Kim, Sang-Yong; Noh, Bong-Soo; Kim, Jung-Hoe; Lee, Jung-Kul

    2003-07-01

    Mutagenesis of Erwinia rhapontici was performed to enhance the production of isomaltulose from sucrose. A mutant strain, BN 68089, was obtained through a screening process involving automated and miniaturized cultivation in Bioscreen C. This high-throughput, miniaturized screening system was optimized to identify the mutant strain, which had a conversion yield (90%) and productivity (194 g l(-1) h(-1)). The BN 68089 mutant cells were immobilized in sodium alginate and when operated in a packed bed reactor gave a yield of 89% and a productivity of 144 g l(-1) h(-1) of at 30 degrees C, the optimal temperature. Immobilized BN 68089 cells exhibited 8% and 15% higher yield and productivity, respectively, than those of the wild-type strain.

  7. Growth and yield of winter wheat (Triticum aestivum L.) and corn (Zea mays L.) near a high voltage transmission line.

    PubMed

    Soja, G; Kunsch, B; Gerzabek, M; Reichenauer, T; Soja, A-M; Rippar, G; Bolhàr-Nordenkampf, H R

    2003-02-01

    The objective of this study was to determine the effects of an electromagnetic field from a high voltage transmission line on the yield of agricultural crops cultivated underneath and near the transmission line. For 5 years, experiments with winter wheat and corn were carried out near the 380 kV transmission line Dürnrohr (Austria)-Slavetice (Czech Republic). Different field strengths were tested by planting the crops at different distances from the transmission line. The plants were grown in experimental plots (1.77 m2), aligned to equal electric field strengths, and were cultivated according to standard agricultural practice. The soil for all plots was homogenized layer-specifically to a depth of 0.5 m to guarantee uniform soil conditions in the plant root environment. The soil was sampled annually for determinations of carbon content and the behavior of microbial biomass. During development of the vegetation, samples were collected at regular intervals for growth rate analyses. At physiological maturity, the plots (n = 8) were harvested for grain and straw yield determinations. The average electric and magnetic field strengths at four distances from the transmission line (nominal distances: 40, 14, 8, and 2 m) were between 0.2 and 4.0 kV/m and between 0.4 and 4.5 micro T, respectively. No effect of the field exposures on soil microbial biomass could be detected. The wheat grain yields were 7% higher (average of 5 years) in the plots with the lowest field exposure than in the plots nearer to the transmission line (P <.10). The responses of the plants were more pronounced in years with drought episodes during grain filling than in humid years. No significant yield differences were found for corn yields. The extent of the yield variations attributed to the distance from the transmission line was small compared to the observed annual variations in climatic or soil specific site characteristics.

  8. A novel plant cell bioproduction platform for high-yield secretion of recombinant proteins.

    PubMed

    Xu, Jianfeng; Kieliszewski, Marcia J

    2012-01-01

    Plant cell suspension culture integrates the merits of whole-plant systems with those of microbial fermentation and mammalian cell culture, and has been recognized as a promising alternative biosynthetic platform for valuable proteins. However, the low protein productivity dilemma has been the bottleneck toward commercializing this technology. Here, we describe a new technology, termed hydroxyproline (Hyp)-Glyco technology, that dramatically increases the yield of secreted recombinant proteins from cultured plant cells by expressing them as fusions with a novel glycomodule tag comprising an Hyp-rich repetitive peptide (HypRP) backbone that is subsequently glycosylated through the Hyp residues. The extensive glycosylation of the HypRP tags greatly extends the serum half-life of small therapeutic proteins, such as interferon α2b or human growth hormone, without significantly impairing their bioactivities and the tag greatly enhances solubility.

  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 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

  11. 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.

  12. Analysis of activated air following high yield shots in the NIF

    DOE PAGES

    Khater, Hesham; Brereton, Sandra

    2015-07-24

    During the ignition experimental campaign, the National Ignition Facility (NIF) is expected to perform shots with varying fusion yield (up to 20 MJ or 7.1 x 1018 neutrons per shot) and a maximum annual yield of 1200 MJ. A detailed MCNP model of the Target Bay (TB) and the two switchyards (SY) has been developed to estimate the post-shot radiation environment inside the facility. During D-T shots, a pulse of 14.1 MeV neutrons streaming outside the Target Chamber (TC) will activate the air present inside the TB and the argon gas inside the laser tubes. Smaller levels of activity aremore » also generated in the SY air and in the argon portion of the SY laser beam path. The activated TB air will be mixed with fresh air from the Operations Support Building (OSB) before release through the stack. Flow of activated air from the Target Bay is controlled by the heating, ventilating, and air conditioning (HVAC) system. 16N (T1/2 = 7.13 s) dominates the radiation levels during the first minute following the shot. It is expected that 16N will decay away during the confinement time before releasing the TB air through the stack. The other major contributors are 13N (T1/2 = 9.97 min) and 41Ar (T1/2 = 1.83 h). In general a low dose rate of < 1 μSv/h is expected near the stack during the first few hours following a 20 MJ shot. Here, the amount of activated Target Bay air released through the stack is very small and does not pose significant hazard to personnel or the environment. In the mean time, due to a very small leakage rate out of the laser tubes, the activated argon gas decays within the tubes and any resulting release to the environment is insignificant.« less

  13. Use of Nonspecific, Glutamic Acid-Free, Media and High Glycerol or High Amylase as Inducing Parameters for Screening Bacillus Isolates Having High Yield of Polyglutamic Acid.

    PubMed

    Baxi, Nandita N

    2014-01-01

    Out of fifty-five Bacillus isolates obtained from ten different regional locations and sources, seven showed the ability to consistently produce specific extracellular polymeric substance (EPS) on rich as well as synthetic but nonspecific media which did not contain glutamic acid. The isolates were identified as either Bacillus licheniformis or Bacillus subtilis. The EPS from all isolates was resistant to alpha protease, proteinase K, and was thus of high molecular weight. Further it was detected after SDS-PAGE by methylene blue but not by coomassie blue R staining as in case of proteins with high proportion of acidic amino acids. Cell-free EPS, after acid hydrolysis, showed absence of carbohydrates and presence of only glutamic acid. Thus the native the EPS from all seven isolates was confirmed to be gamma polyglutamic acid (PGA) and not exopolysaccharide. The Bacillus isolate T which produced maximum polymer on all media tested had higher amylase: protease activity as compared to other strains. If inoculum was developed in rich medium as compared to synthetic medium, the PGA produced increased by twofold in the subsequent synthetic production medium. Similarly, use of inoculum consisting of young and vegetative cells also increased the PGA production by twofold though amount of inoculum did not affect yield of PGA. Though PGA was produced in even in the absence of glutamic acid supplementation in the production medium by all isolates, the yield of PGA increased by fourfold in the presence glutamic acid and the maximum yield was 30 g/l for isolate K. The supplementation of glutamine instead of glutamic acid into the medium caused an increase in the viscosity of the non-Newtonian solution of PGA.

  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. Development and characterization of a high yield transportable pulsed neutron source with efficient and compact pulsed power system.

    PubMed

    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 × 10(9) 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.

  17. Quantum-orbit analysis for yield and ellipticity of high order harmonic generation with elliptically polarized laser field.

    PubMed

    Li, Yang; Zhu, Xiaosong; Zhang, Qingbin; Qin, Meiyan; Lu, Peixiang

    2013-02-25

    We perform a quantum-orbit analysis for the dependence of high-order-harmonic yield on the driving field ellipticity and the polarization properties of the generated high harmonics. The electron trajectories responsible for the emission of particular harmonics are identified. It is found that, in elliptically polarized driving field, the electrons have ellipticity-dependent initial velocities, which lead to the decrease of the ionization rate. Thus the harmonic yield steeply decreases with laser ellipticity. Besides, we show that the polarization properties of the harmonics are related to the complex momenta of the electron. The physical origin of the harmonic ellipticity is interpreted as the consequence of quantum-mechanical uncertainty of the electron momentum. Our results are verified with the experimental results as well as the numerical solutions of the time dependent Schrödinger equation from the literature.

  18. Enhancement of astaxanthin production from Haematococcus pluvialis mutants by three-stage mutagenesis breeding.

    PubMed

    Wang, Ni; Guan, Bin; Kong, Qing; Sun, Han; Geng, Zhaoyan; Duan, Liangfei

    2016-10-20

    Haematococcus pluvialis was modified for higher astaxanthin production compatible with the superiorities of high biomass and high activity by three-stage mutagenesis breeding. UV irradiation mutants named UV11-4 made an increase on cell dry weight, but showed a longer growth circle than the wild type. On the basis of UV mutants, ethyl methane sulphonate (EMS) mutants E2-5 cut down the latent phase, brought forward and extended the logarithmic phase. The inhibitor diphenylamine (DPA) was employed to screen high-yield astaxanthin producer by the color change of colonies from green to red on solid medium. Via the contravariant cultivation, proliferation and transformation, the mutant DPA12-2 possessed an 1.7-fold astaxanthin production compared to the wild type, reaching 47.21±3.30mg/g dry cells.

  19. Organocatalytic azomethine imine-olefin click reaction: high-yielding stereoselective synthesis of spiroindane-1,3-dione-pyrazolidinones.

    PubMed

    Ramachary, Dhevalapally B; Prabhakar Reddy, T; Suresh Kumar, A

    2016-07-06

    In search of developing new useful "click reactions", herein we report the organocatalytic azomethine imine-olefin [3 + 2]-cycloaddition as a new click reaction for the synthesis of drug-like spiroindane-1,3-dione-pyrazolidinones from indane-1,3-diones, aldehydes and N,N-cyclic azomethine imines through amino acid-catalysis. The scope of this new click reaction is demonstrated using many examples with high reactivity, selectivity and yields.

  20. Heating of proteins as a means of improving crystallization: a successful case study on a highly amyloidogenic triple mutant of human transthyretin

    SciTech Connect

    Karlsson, Anders; Sauer-Eriksson, A. Elisabeth

    2007-08-01

    By heating a highly amyloidogenic mutant of the human plasma protein transthyretin at 328 K for 48 h, diffraction-quality crystals could be reproducibly produced. The procedure precipitated ∼40% of the protein, but rendered what remained in solution more homogenous. The use of high temperatures in the purification procedures of heat-stable proteins is a well established technique. Recently, rapid pre-heat treatment of protein samples prior to crystallization trials was described as a final polishing step to improve the diffraction properties of crystals [Pusey et al. (2005 ▶), Prog. Biophys. Mol. Biol. 88, 359–386]. The present study demonstrates that extended high-temperature incubation (328 K for 48 h) of the highly amyloidogenic transthyretin mutant TTR G53S/E54D/L55S successfully removes heterogeneities and allows the reproducible growth of well diffracting crystals. Heat treatment might be applied as an optimization method to other cases in which the protein/biomolecule fails to form diffracting crystals.

  1. Analysis of activated air following high yield shots in the NIF

    SciTech Connect

    Khater, Hesham; Brereton, Sandra

    2015-07-24

    During the ignition experimental campaign, the National Ignition Facility (NIF) is expected to perform shots with varying fusion yield (up to 20 MJ or 7.1 x 1018 neutrons per shot) and a maximum annual yield of 1200 MJ. A detailed MCNP model of the Target Bay (TB) and the two switchyards (SY) has been developed to estimate the post-shot radiation environment inside the facility. During D-T shots, a pulse of 14.1 MeV neutrons streaming outside the Target Chamber (TC) will activate the air present inside the TB and the argon gas inside the laser tubes. Smaller levels of activity are also generated in the SY air and in the argon portion of the SY laser beam path. The activated TB air will be mixed with fresh air from the Operations Support Building (OSB) before release through the stack. Flow of activated air from the Target Bay is controlled by the heating, ventilating, and air conditioning (HVAC) system. 16N (T1/2 = 7.13 s) dominates the radiation levels during the first minute following the shot. It is expected that 16N will decay away during the confinement time before releasing the TB air through the stack. The other major contributors are 13N (T1/2 = 9.97 min) and 41Ar (T1/2 = 1.83 h). In general a low dose rate of < 1 μSv/h is expected near the stack during the first few hours following a 20 MJ shot. Here, the amount of activated Target Bay air released through the stack is very small and does not pose significant hazard to personnel or the environment. In the mean time, due to a very small leakage rate out of the laser tubes, the activated argon gas decays within the tubes and any resulting release to the environment is insignificant.

  2. 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

  3. 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.

  4. Understanding scaling of ignition metrics for high-yield implosions on the NIF

    NASA Astrophysics Data System (ADS)

    Springer, Paul; Hurricane, Omar; Hammer, J. H.; Callahan, D. A.; Casey, D. T.; Cerjan, C. J.; Edwards, M. J.; Field, J. E.; Gaffney, J.; Grim, G. P.; Kritcher, A. L.; Ma, T.; Macphee, A. G.; Munro, D. H.; Nora, R. C.; Patel, P. K.; Peterson, L.; Spears, B.

    2016-10-01

    The self-heating condition for an imploding hotspot requires understanding the balance between mechanical work, heating via fusion reactions, and the radiative and conduction losses. A 3D cognizant Lawson ignition threshold metric is derived based on net fusion hotspot heating achieved when hotspot rho-r and ion temperature exceed critical values that depend on the temperature-dependent loss mechanisms. Key to understanding and scaling such analysis is an accurate determination of hotspot density and pressure, which are generally inferred using the yield, the thermal temperature, and other experimental data. 3D flow and its effect on neutron spectra can lead to overestimation of the temperature, and underestimation of hotspot rho-r, energy, and ignition margin. In this work, we analyze these effects in NIF data, and propose new methods to avoid them. These simple, analytical methods are tested using the largest 2D ICF simulation dataset ever produced. Work performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under contract DE-AC52-07NA273.

  5. 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.

  6. 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.

  7. A mercury-catalyzed, high-yield system for the oxidation of methane to methanol.

    PubMed

    Periana, R A; Taube, D J; Evitt, E R; Löffler, D G; Wentrcek, P R; Voss, G; Masuda, T

    1993-01-15

    A homogeneous system for the selective, catalytic oxidation of methane to methanol via methyl bisulfate is reported. The net reaction catalyzed by mercuric ions, Hg(II), is the oxidation of methane by concentrated sulfuric acid to produce methyl bisulfate, water, and sulfur dioxide. The reaction is efficient. At a methane conversion of 50 percent, 85 percent selectivity to methyl bisulfate ( approximately 43 percent yield; the major side product is carbon dioxide) was achieved at a molar productivity of 10(-7) mole per cubic centimeter per second and Hg(II) turnover frequency of 10(-3) per second. Separate hydrolysis of methyl bisulfate and reoxidation of the sulfur dioxide with air provides a potentially practical scheme for the oxidation of methane to methanol with molecular oxygen. The primary steps of the Hg(II)-catalyzed reaction were individually examined and the essential elements of the mechanism were identified. The Hg(II) ion reacts with methane by an electrophilic displacement mechanism to produce an observable species, CH(3)HgOSO(3)H, 1. Under the reaction conditions, 1 readily decomposes to CH(3)OSO(3)H and the reduced mercurous species, Hg(2)(2+) The catalytic cycle is completed by the reoxidation of Hg(2)(2+) with H(2)SO(4) to regenerate Hg(II) and byproducts SO(2) and H(2)O. Thallium(III), palladium(II), and the cations of platinum and gold also oxidize methane to methyl bisulfate in sulfuric acid.

  8. A Mercury-Catalyzed, High-Yield System for the Oxidation of Methane to Methanol

    NASA Astrophysics Data System (ADS)

    Periana, Roy A.; Taube, Douglas J.; Evitt, Eric R.; Loffler, Daniel G.; Wentrcek, Paul R.; Voss, George; Masuda, Toshihiko

    1993-01-01

    A homogeneous system for the selective, catalytic oxidation of methane to methanol via methyl bisulfate is reported. The net reaction catalyzed by mercuric ions, Hg(II), is the oxidation of methane by concentrated sulfuric acid to produce methyl bisulfate, water, and sulfur dioxide. The reaction is efficient. At a methane conversion of 50 percent, 85 percent selectivity to methyl bisulfate (~43 percent yield; the major side product is carbon dioxide) was achieved at a molar productivity of 10-7 mole per cubic centimeter per second and Hg(II) turnover frequency of 10-3 per second. Separate hydrolysis of methyl bisulfate and reoxidation of the sulfur dioxide with air provides a potentially practical scheme for the oxidation of methane to methanol with molecular oxygen. The primary steps of the Hg(II)-catalyzed reaction were individually examined and the essential elements of the mechanism were identified. The Hg(II) ion reacts with methane by an electrophilic displacement mechanism to produce an observable species, CH_3HgOSO_3H, 1. Under the reaction conditions, 1 readily decomposes to CH_3OSO_3H and the reduced mercurous species, Hg_22+. The catalytic cycle is completed by the reoxidation of Hg_22+ with H_2SO_4 to regenerate Hg(II) and byproducts SO_2 and H_2O. Thallium(III), palladium(II), and the cations of platinum and gold also oxidize methane to methyl bisulfate in sulfuric acid.

  9. 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.

  10. 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.

  11. Discrete Electronic Bands in Semiconductors and Insulators: Potential High-Light-Yield Scintillators

    DOE PAGES

    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

  12. 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.

  13. Screening of High-Level 4-Hydroxy-2 (or 5)-Ethyl-5 (or 2)-Methyl-3(2H)-Furanone-Producing Strains from a Collection of Gene Deletion Mutants of Saccharomyces cerevisiae

    PubMed Central

    Watanabe, Jun; Akao, Takeshi; Watanabe, Daisuke; Mogi, Yoshinobu; Shimoi, Hitoshi

    2014-01-01

    4-Hydroxy-2 (or 5)-ethyl-5 (or 2)-methyl-3(2H)-furanone (HEMF) is an important flavor compound that contributes to the sensory properties of many natural products, particularly soy sauce and soybean paste. The compound exhibits a caramel-like aroma and several important physiological activities, such as strong antioxidant activity. HEMF is produced by yeast species in soy sauce manufacturing; however, the enzymes involved in HEMF production remain unknown, hindering efforts to breed yeasts with high-level HEMF production. In this study, we identified high-level HEMF-producing mutants among a Saccharomyces cerevisiae gene deletion mutant collection. Fourteen deletion mutants were screened as high-level HEMF-producing mutants, and the ADH1 gene deletion mutant (adh1Δ) exhibited the maximum HEMF production capacity. Further investigations of the adh1Δ mutant implied that acetaldehyde accumulation contributes to HEMF production, agreeing with previous findings. Therefore, acetaldehyde might be a precursor for HEMF. The ADH1 gene deletion mutant of Zygosaccharomyces rouxii, which is the dominant strain of yeast found during soy sauce fermentation, also produces HEMF effectively, suggesting that acetaldehyde accumulation might be a benchmark for breeding industrial yeasts with excellent HEMF production abilities. PMID:25362059

  14. 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.

  15. 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.

  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. 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.

  18. High Yield Expression of Recombinant Human Proteins with the Transient Transfection of HEK293 Cells in Suspension

    PubMed Central

    Subedi, Ganesh P.; Johnson, Roy W.; Moniz, Heather A.; Moremen, Kelley W.; Barb, Adam

    2015-01-01

    The art of producing recombinant proteins with complex post-translational modifications represents a major challenge for studies of structure and function. The rapid establishment and high recovery from transiently-transfected mammalian cell lines addresses this barrier and is an effective means of expressing proteins that are naturally channeled through the ER and Golgi-mediated secretory pathway. Here is one protocol for protein expression using the human HEK293F and HEK293S cell lines transfected with a mammalian expression vector designed for high protein yields. The applicability of this system is demonstrated using three representative glycoproteins that expressed with yields between 95-120 mg of purified protein recovered per liter of culture. These proteins are the human FcγRIIIa and the rat α2-6 sialyltransferase, ST6GalI, both expressed with an N-terminal GFP fusion, as well as the unmodified human immunoglobulin G1 Fc. This robust system utilizes a serum-free medium that is adaptable for expression of isotopically enriched proteins and carbohydrates for structural studies using mass spectrometry and nuclear magnetic resonance spectroscopy. Furthermore, the composition of the N-glycan can be tuned by adding a small molecule to prevent certain glycan modifications in a manner that does not reduce yield. PMID:26779721

  19. Reassortment of high-yield influenza viruses in vero cells and safety assessment as candidate vaccine strains.

    PubMed

    Zhou, Jian; Yang, Fan; Yang, Jinghui; Ma, Lei; Cun, Yina; Song, Shaohui; Liao, Guoyang

    2017-01-02

    Vaccination is the practiced and accessible measure for preventing influenza infection. Because chicken embryos used for vaccine production have various insufficiencies, more efficient methods are needed. African green monkey kidney (Vero) cells are recommended by the World Health Organization (WHO) as a safe substitute for influenza vaccine production for humans. However, the influenza virus usually had low-yield in Vero cells, which limits the usage of Vero cellular vaccines. This study used 2 high-yield influenza viruses in Vero cells: A/Yunnan/1/2005Va (H3N2) and B/Yunnan/2/2005Va (B) as donor viruses. It used 3 wild strain viruses to reassort new adaptation viruses, including: A/Tianjin/15/2009(H1N1), A/Fujian/196/2009(H3N2), and B/Chongqing/1384/2010(B). These three new viruses could maintain the characteristic of high-yield in Vero cells. Furthermore, they could keep the immunogenic characteristics of the original wild influenza viruses. Importantly, these viruses were shown as safe in chicken embryo and guinea pigs assessment systems. These results provide an alternative method to produce influenza vaccine based on Vero cells.

  20. Endophytic Fungus Nigrospora oryzae from a Medicinal plant Coccinia grandis, a High Yielding New Source of Phenazine-1-carboxamide.

    PubMed

    Thanabalasingam, Dharushana; Kumar, N Savitri; Jayasinghe, Lalith; Fujimoto, Yoshinori

    2015-10-01

    Nigrospora oryzae was isolated as an endophytic fungus from the leaves of Coccinia grandis, a popular medicinal plant used to control diabetes. Fermentation of the fungus in potato dextrose broth and chromatographic purification of the ethyl acetate extracts of the broth and mycelium yielded two phenazine secondary metabolites, which were identified as phenazine-1-carboxylic acid (1) and phenazine-1-carboxamide (2) by comparing their spectral data with those reported in the literature. Compound 2, isolated in high yield (1 g/4 L medium), showed strong antifungal activity against the plant pathogen Cladosporium cladosporioides. This is the first report of the isolation of N. oryzae as an endophytic fungus of C. grandis. These phenazines have never been isolated from any fungal source. Antifungal activity of 2 against C. cladosporioides is reported for the first time.

  1. Yield Strength of Transparent MgAl2O4 Nano-Ceramic at High Pressure and Temperature.

    PubMed

    Zhang, Jie; Lu, Tiecheng; Chang, Xianghui; Jiang, Shengli; Wei, Nian; Qi, Jianqi

    2010-05-23

    We report here experimental results of yield strength and stress relaxation measurements of transparent MgAl2O4 nano-ceramics at high pressure and temperature. During compression at ambient temperature, the differential strain deduced from peak broadening increased significantly with pressure up to 2 GPa, with no clear indication of strain saturation. However, by then, warming the sample above 400°C under 4 GPa, stress relaxation was obviously observed, and all subsequent plastic deformation cycles are characterized again by peak broadening. Our results reveal a remarkable reduction in yield strength as the sintering temperature increases from 400 to 900°C. The low temperature for the onset of stress relaxation has attracted attention regarding the performance of transparent MgAl2O4 nano-ceramics as an engineering material.

  2. Development of an FPW Biosensor with Low Insertion Loss and High Fabrication Yield for Detection of Carcinoembryonic Antigen

    PubMed Central

    Lan, Je-Wei; Huang, I-Yu; Lin, Yu-Cheng; Lin, Chang-Yu; Chen, Jian-Lin; Hsieh, Chia-Hsu

    2016-01-01

    In the last two decades, various flexural plate-wave (FPW)-based biosensors with low phase velocity, low operation frequency, high sensitivity, and short response time, have been developed. However, conventional FPW transducers have low fabrication yield because controlling the thickness of silicon/isolation/metal/piezoelectric multilayer floating thin-plate is difficult. Additionally, conventional FPW devices usually have high insertion loss because of wave energy dissipation to the silicon substrate or outside area of the output interdigital transducers (IDTs). These two disadvantages hinder the application of FPW devices. To reduce the high insertion loss of FPW devices, we designed two focus-type IDTs (fan-shaped and circular, respectively) that can effectively confine the launched wave energy, and adopted a focus-type silicon-grooved reflective grating structure (RGS) that can reduce the wave propagation loss. To accurately control the thickness of the silicon thin-plate and substantially improve the fabrication yield of FPW transducers, a 60 °C/27 °C two-step anisotropic wet etching process was developed. Compared with conventional FPW devices (with parallel-type IDTs and without RGS), the proposed FPW devices have lower insertion loss (36.04 dB) and higher fabrication yield (63.88%). Furthermore, by using cystamine-based self-assembled monolayer (SAM) nanotechnology, we used the improved FPW device to develop a novel FPW-based carcinoembryonic antigen (CEA) biosensor for detection of colorectal cancer, and this FPW-CEA biosensor has a low detection limit (5 ng/mL), short response time (<10 min), high sensitivity (60.16–70.06 cm2/g), and high sensing linearity (R-square = 0.859–0.980). PMID:27834798

  3. Evaluating low lignin mutants of forage sorghum for increased conversion efficiency to sugars and ethanol

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Reduced lignin near-isogenic lines of Atlas bmr-6, bmr-12, and bmr-6 bmr-12 forage sorghum (Sorghum biocolor (L.)) were evaluated as sources of biomass for conversion to sugars and ethanol. These mutants have the advantage of reduced lignin contents and high biomass yields. Field replicates of wil...

  4. 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

  5. High-Yield Endoglucanase Production by Trichoderma harzianum IOC-3844 Cultivated in Pretreated Sugarcane Mill Byproduct.

    PubMed

    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

    2010-09-26

    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.

  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 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

  7. Lustrous copper nanoparticle film: Photodeposition with high quantum yield and electric conductivity

    NASA Astrophysics Data System (ADS)

    Miyagawa, Masaya; Yonemura, Mari; Tanaka, Hideki

    2016-11-01

    Cu nanoparticle (NP) film has attracted much attention due to its high electric conductivity. In the present study, we prepared a Cu NP film on a TiO2-coated substrate by photoreduction of copper acetate solution. The obtained film showed high electric conductivity and metallic luster by the successive deposition of Cu NP. Moreover, the film was decomposed on exposure to fresh air, and its decomposition reaction mechanisms were proposed. Hence, we concluded that the obtained lustrous film was composed of Cu NP, even though its physical properties was similar to bulk copper.

  8. Maximizing the yield and cutoff of high-order harmonic generation from plasma plume

    SciTech Connect

    Ganeev, Rashid A.; Elouga Bom, Luc B.; Ozaki, Tsuneyuki; Redkin, Pavel V.

    2007-11-15

    We study high-order harmonic generation (HHG) from various lowly ionized laser plasmas. We study harmonic generation from targets of Al (Z=13) to Bi (Z=83). Varying the wavelength, chirp, and pulse duration of the femtosecond pump laser resulted in the change in the harmonic distribution, cutoff, and conversion efficiency of HHG. We also study the use of doubly charged ions, and resonances for some materials. We were able to obtain high HHG conversion efficiency and harmonic cutoff by implementing the above approaches and by observing the time-resolved spectra of the laser plasma.

  9. High-level expression and characterization of a thermostable xylanase mutant from Trichoderma reesei in Pichia pastoris.

    PubMed

    Li, Yang-yuan; Zhong, Kai-xin; Hu, Ai-hong; Liu, Dan-ni; Chen, Li-zhi; Xu, Shu-de

    2015-04-01

    A gene encoding xylanase 2 mutant from Trichoderma reesei (T2C/T28C, named mxyn2) was cloned into the Pichia pastoris X33 strain using the vector pPICZαA. Recombinant Mxyn2p was functionally expressed in P. pastoris X33 and secreted into the supernatant. Real time qPCR demonstrated that an increase in gene copy number correlated with higher levels of expression. Supernatant from methanol induced cells was concentrated by ultrafiltration with a 10kDa cut off membrane, and purified with ion exchange chromatography using SP Sepharose Fast Flow chromatography. Recombinant Mxyn2p protein had the highest activity at 75°C, while recombinant protein encoded by the "wild type" xylanase gene xyn2, also expressed in Pichia, was 20°C lower. The Mxyn2p enzyme retained more than 70% of its activity after incubation at 80°C for 10min. The effects of the optimal pH and temperature for higher expression levels in P. pastoris were also determined, 6.0 and 22°C, respectively. The maximum xylanase activity of Mxyn2p was 13,000nkat/mg (9.88g/l) in fed-batch cultivation after 168h induction with methanol in a 50l bioreactor.

  10. Genome-wide Hi-C analyses in wild type and mutants reveal high-resolution chromatin interactions in Arabidopsis

    PubMed Central

    Feng, Suhua; Cokus, Shawn J.; Schubert, Veit; Zhai, Jixian; Pellegrini, Matteo; Jacobsen, Steven E.

    2015-01-01

    SUMMARY Chromosomes form three-dimensional structures that are critical to the regulation of cellular and genetic processes. Here, we present a study of global chromatin interaction patterns in Arabidopsis thaliana. Our genome-wide approach confirmed interactions that were previously observed by other methods as well as uncovered previously unknown long-range interactions such as those among small heterochromatic regions embedded in euchromatic arms. We also found that interactions are correlated with various epigenetic marks that are localized in active or silenced chromatin. Arabidopsis chromosomes do not contain large local interactive domains that resemble the topological domains described in animals, but instead contain relatively small interactive regions scattered around the genome that contain H3K27me3 or H3K9me2. We generated interaction maps in mutants that are defective in specific epigenetic pathways and found altered interaction patterns that correlate with changes in the epigenome. These analyses provide further insights into molecular mechanisms of epigenetic regulation of the genome. PMID:25132175

  11. 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.

  12. The lux genes of the luminous bacterial symbiont, Photobacterium leiognathi, of the ponyfish. Nucleotide sequence, difference in gene organization, and high expression in mutant Escherichia coli.

    PubMed

    Lee, C Y; Szittner, R B; Meighen, E A

    1991-10-01

    The lux genes required for light expression in the luminescent bacterium Photobacterium leiognathi (ATCC 25521) have been cloned and expressed in Escherichia coli and their organization and nucleotide sequence determined. Transformation of a recombinant 9.5-kbp chromosomal DNA fragment of P. leiognathi into an E. coli mutant (43R) gave luminescent colonies that were as bright as those of the parental strain. Moreover, expression of the lux genes in the mutant E. coli was strong enough so that not only were high levels of luciferase detected in crude extracts, but the fatty-acid reductase activity responsible for synthesis of the aldehyde substrate for the luminescent reaction could readily be measured. Determination of the 7.3-kbp nucleotide sequence of P. leiognathi DNA, including the genes for luciferase (luxAB) and fatty-acid reductase (luxCDE) as well as a new lux gene (luxG) found recently in luminescent Vibrio species, showed that the order of the lux genes was luxCDABEG. Moreover, luxF, a gene homologous to luxB and located between luxB and luxE in Photobacterium but not Vibrio strains, was absent. In spite of this different lux gene organization, an intergenic stem-loop structure between luxB and luxE was discovered to be highly conserved in other Photobacterium species after luxF.

  13. 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

    2016-01-04

    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.

  14. Steady-state and time-resolved fluorescence studies on wild type and mutant chromatium vinosum high potential iron proteins: holo- and apo-forms.

    PubMed Central

    Sau, A K; Chen, C A; Cowan, J A; Mazumdar, S; Mitra, S

    2001-01-01

    Detailed circular dichroism (CD), steady-state and time-resolved tryptophan fluorescence studies on the holo- and apo- forms of high potential iron protein (HiPIP) from Chromatium vinosum and its mutant protein have been carried out to investigate conformational properties of the protein. CD studies showed that the protein does not have any significant secondary structure elements in the holo- or apo- HiPIP, indicating that the metal cluster does not have any effect on formation of secondary structure in the protein. Steady-state fluorescence quenching studies however, suggested that removal of the iron-sulfur ([Fe(4)S(4)](3+)) cluster from the protein leads to an increase in the solvent accessibility of tryptophans, indicating change in the tertiary structure of the protein. CD studies on the holo- and apo- HiPIP also showed that removal of the metal prosthetic group drastically affects the tertiary structure of the protein. Time-resolved fluorescence decay of the wild type protein was fitted to a four-exponentials model and that of the W80N mutant was fitted to a three-exponentials model. The time-resolved fluorescence decay was also analyzed by maximum entropy method (MEM). The results of the MEM analysis agreed with those obtained from discrete exponentials model analysis. Studies on the wild type and mutants helped to assign the fast picosecond lifetime component to the W80 residue, which exhibits fast fluorescence energy transfer to the [Fe(4)S(4)](3+) cluster of the protein. Decay-associated fluorescence spectra of each tryptophan residues were calculated from the time-resolved fluorescence results at different emission wavelengths. The results suggested that W80 is in the hydrophobic core of the protein, but W60 and W76 are partially or completely exposed to the solvent. PMID:11566801

  15. 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

  16. A high yield reverse micelle synthesis of catalysts and catalyst precursors

    SciTech Connect

    Linehan, J.C.; Matson, D.W.; Darab, J.G.

    1995-12-01

    Water-in-oil (w/o) microemulsions containing high ionic strengths in their aqueous cores have been produced by using a stabilizing co-surfactant. These {open_quotes}modified{close_quotes} w/o microemulsions have proven to be effective media in which to synthesize nanophase metal, metal oxide, metal sulfide, and mixed metal containing powders suitable for use as catalysts or catalyst precursors. Up to 10 grams of nanocrystalline powder can be produced per liter of the modified microemulsion because of the high reactant concentrations that can be attained using these systems. The nanocrystalline powders produced have been characterized by XRD, SEM, and TEM, as well as by other techniques where appropriate.

  17. 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

  18. High yield hydrogen production in a single-chamber membrane-less microbial electrolysis cell.

    PubMed

    Ye, Yejie; Wang, Liyong; Chen, Yingwen; Zhu, Shemin; Shen, Shubao

    2010-01-01

    The single-chamber membrane-less MEC exerted much better hydrogen production performance while given higher applied voltages than it did at lower. High applied voltages that could shorten the reaction time and the exposure of anode to air for at least 30 min between cycles can significantly suppress methanogen and increase hydrogen production. At an applied voltage of 1.0 V, a hydrogen production rate of 1.02 m(3)/m(3)/day with a current density of 5.7 A/m(2) was achieved. Cathodic hydrogen recovery and coulombic efficiency were 63.4% and 69.3% respectively. The hydrogen concentration of mixture gas produced of 98.4% was obtained at 1.0 V, which was the best result of reports. The reasons that such a high hydrogen concentration can be achieved were probably the high electrochemical activity and hydrogen production capability of the active microorganisms. Increase in substrate concentrations could not improve MEC's performance, but increased the reaction times. Further, reactor configuration and operation factors optimisation should be considered to increase current density, hydrogen production rate and hydrogen recovery.

  19. High Yields of Hydrogen Production Induced by Meta-Substituted Dichlorophenols Biodegradation from the Green Alga Scenedesmus obliquus

    PubMed Central

    Papazi, Aikaterini; Andronis, Efthimios; Ioannidis, Nikolaos E.; Chaniotakis, Nikolaos; Kotzabasis, Kiriakos

    2012-01-01

    Hydrogen is a highly promising energy source with important social and economic implications. The ability of green algae to produce photosynthetic hydrogen under anaerobic conditions has been known for years. However, until today the yield of production has been very low, limiting an industrial scale use. In the present paper, 73 years after the first report on H2-production from green algae, we present a combinational biological system where the biodegradation procedure of one meta-substituted dichlorophenol (m-dcp) is the key element for maintaining continuous and high rate H2-production (>100 times higher than previously reported) in chloroplasts and mitochondria of the green alga Scenedesmus obliquus. In particular, we report that reduced m-dcps (biodegradation intermediates) mimic endogenous electron and proton carriers in chloroplasts and mitochondria, inhibit Photosystem II (PSII) activity (and therefore O2 production) and enhance Photosystem I (PSI) and hydrogenase activity. In addition, we show that there are some indications for hydrogen production from sources other than chloroplasts in Scenedesmus obliquus. The regulation of these multistage and highly evolved redox pathways leads to high yields of hydrogen production and paves the way for an efficient application to industrial scale use, utilizing simple energy sources and one meta-substituted dichlorophenol as regulating elements. PMID:23145057

  20. Bioconversion of glycerol to ethanol by a mutant Enterobacter aerogenes.

    PubMed

    Nwachukwu, Res; Shahbazi, A; Wang, L; Ibrahim, S; Worku, M; Schimmel, K

    2012-03-29

    The main objective of this research is to develop, by adaptive evolution, mutant strains of Enterobacter aerogenes ATCC 13048 that are capable of withstanding high glycerol concentration as well as resisting ethanol-inhibition. The mutant will be used for high ethanol fermentation from glycerol feedstock. Ethanol production from pure (P-) and recovered (R-) glycerol using the stock was evaluated. A six-tube-subculture-generations method was used for developing the mutant. This involved subculturing the organism six consecutive times in tubes containing the same glycerol and ethanol concentrations at the same culture conditions. Then, the glycerol and/or ethanol concentration was increased and the six subculture generations were repeated. A strain capable of growing in 200 g/L glycerol and 30 g/L ethanol was obtained. The ability of this mutant, vis-à-vis the original strain, in utilizing glycerol in a high glycerol containing medium, with the concomitant ethanol yield, was assessed. Tryptic soy broth without dextrose (TSB) was used as the fermentation medium. Fermentation products were analyzed using HPLC.In a 20 g/L glycerol TSB, E. aerogenes ATCC 13048 converted 18.5 g/L P-glycerol and 17.8 g/L R-glycerol into 12 and 12.8 g/L ethanol, respectively. In a 50 g/L P-glycerol TSB, it utilized only 15.6 g/L glycerol; but the new strain used up 39 g/L, yielding 20 g/L ethanol after 120 h, an equivalence of 1.02 mol ethanol/mol-glycerol. This is the highest ethanol yield reported from glycerol bioconversion. The result of this P-glycerol fermentation can be duplicated using the R-glycerol from biodiesel production.

  1. High-yield isolation of extracellular vesicles using aqueous two-phase system

    PubMed Central

    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

  2. A high yield reverse micelle synthesis of catalysts and catalyst precursors

    SciTech Connect

    Linehan, J.C.; Matson, D.W.; Darab, J.G.; Fulton, J.L.

    1995-04-01

    Reverse micelles or water-in-oil microemulsions have been prepared using a mixed AOT/SDS surfactant to increase the stability of the microemulsion and thereby allow a high loading of particle-forming precursors in the aqueous cores. The Modified Reverse Micelles (MRM), as these new binary surfactant microemulsions are called, have proven useful for the laboratory-scale synthesis of nanoscale metals, metal oxides, metal sulfides, and mixed metal materials. The system allows control over the phase and size of the precipitated crystallites and is ideal for producing nanocrystalline powders and suspensions.

  3. High-yield synthesis of brookite TiO.sub.2 nanoparticles

    SciTech Connect

    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.

  4. 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.

  5. Safe, Selective, and High-Yielding Synthesis of Acryloyl Chloride in a Continuous-Flow System.

    PubMed

    Movsisyan, Marine; Heugebaert, Thomas S A; Dams, Rudy; Stevens, Christian V

    2016-08-09

    Acid chlorides are an important class of compounds and their high reactivity and instability has prompted us to develop a straightforward procedure for their synthesis with on-demand and on-site synthesis possibilities. The focus of this report is acryloyl chloride, mainly important for the acrylate and polymer industry. A continuous-flow methodology was developed for the fast and selective synthesis of the otherwise highly unstable acryloyl chloride. Three routes were investigated in a microreactor setup and all three can potentially be used for its production. The methodology was further expanded to the synthesis of other unstable acid chlorides by both the thionyl chloride and the oxalyl chloride mediated processes. The most sustainable method was the oxalyl chloride mediated procedure under solvent-free conditions, in which near-equimolar amounts of carboxylic acid and oxalyl chloride were used in the presence of catalytic amounts of DMF at room temperature. Within 1 to 3 min, nearly full conversions into the acid chlorides were achieved.

  6. A comparison of continuum and kinetic simulations of microplasmas integrated with high secondary yield cathodes

    NASA Astrophysics Data System (ADS)

    Alamatsaz, Arghavan; Verma, Abhishek Kumar; Venkattraman, Ayyaswamy

    2016-10-01

    During the last two decades, microplasmas have become an active area of research in the field of low-temperature plasma science and engineering with a wide range of applications including electronics, nanomaterial synthesis and metamaterials to name a few. Kinetic and continuum methods are commonly employed numerical simulation techniques to study the low temperature plasmas. The uncertainty and imprecision associated with input parameters used in these models impose a constraint on fidelity of the simulation results. In this work, these computational techniques are compared in the context of modeling microplasmas driven