Sample records for mutant h184f exhibited
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Diminished self-chaperoning activity of the DeltaF508 mutant of CFTR results in protein misfolding.
Directory of Open Access Journals (Sweden)
Adrian W R Serohijos
2008-02-01
Full Text Available The absence of a functional ATP Binding Cassette (ABC protein called the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR from apical membranes of epithelial cells is responsible for cystic fibrosis (CF. Over 90% of CF patients carry at least one mutant allele with deletion of phenylalanine at position 508 located in the N-terminal nucleotide binding domain (NBD1. Biochemical and cell biological studies show that the DeltaF508 mutant exhibits inefficient biosynthetic maturation and susceptibility to degradation probably due to misfolding of NBD1 and the resultant misassembly of other domains. However, little is known about the direct effect of the Phe508 deletion on the NBD1 folding, which is essential for rational design strategies of cystic fibrosis treatment. Here we show that the deletion of Phe508 alters the folding dynamics and kinetics of NBD1, thus possibly affecting the assembly of the complete CFTR. Using molecular dynamics simulations, we find that meta-stable intermediate states appearing on wild type and mutant folding pathways are populated differently and that their kinetic accessibilities are distinct. The structural basis of the increased misfolding propensity of the DeltaF508 NBD1 mutant is the perturbation of interactions in residue pairs Q493/P574 and F575/F578 found in loop S7-H6. As a proof-of-principle that the S7-H6 loop conformation can modulate the folding kinetics of NBD1, we virtually design rescue mutations in the identified critical interactions to force the S7-H6 loop into the wild type conformation. Two redesigned NBD1-DeltaF508 variants exhibited significantly higher folding probabilities than the original NBD1-DeltaF508, thereby partially rescuing folding ability of the NBD1-DeltaF508 mutant. We propose that these observed defects in folding kinetics of mutant NBD1 may also be modulated by structures separate from the 508 site. The identified structural determinants of increased misfolding propensity of
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Cleanup Verification Package for the 126-F-1, 184-F Powerhouse Ash Pit
International Nuclear Information System (INIS)
Clark, S.W.; Sulloway, H.M.
2007-01-01
This cleanup verification package documents completion of remedial action for the 126-F-1, 184-F Powerhouse Ash Pit. This waste site received coal ash from the 100-F Area coal-fired steam plant. Leakage of process effluent from the 116-F-14 , 107-F Retention Basins flowed south into the ash pit, contaminating the northern portion
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Diabetes and exocrine pancreatic insufficiency in E2F1/E2F2 double-mutant mice.
Iglesias, Ainhoa; Murga, Matilde; Laresgoiti, Usua; Skoudy, Anouchka; Bernales, Irantzu; Fullaondo, Asier; Moreno, Bernardino; Lloreta, José; Field, Seth J; Real, Francisco X; Zubiaga, Ana M
2004-05-01
E2F transcription factors are thought to be key regulators of cell growth control. Here we use mutant mouse strains to investigate the function of E2F1 and E2F2 in vivo. E2F1/E2F2 compound-mutant mice develop nonautoimmune insulin-deficient diabetes and exocrine pancreatic dysfunction characterized by endocrine and exocrine cell dysplasia, a reduction in the number and size of acini and islets, and their replacement by ductal structures and adipose tissue. Mutant pancreatic cells exhibit increased rates of DNA replication but also of apoptosis, resulting in severe pancreatic atrophy. The expression of genes involved in DNA replication and cell cycle control was upregulated in the E2F1/E2F2 compound-mutant pancreas, suggesting that their expression is repressed by E2F1/E2F2 activities and that the inappropriate cell cycle found in the mutant pancreas is likely the result of the deregulated expression of these genes. Interestingly, the expression of ductal cell and adipocyte differentiation marker genes was also upregulated, whereas expression of pancreatic cell marker genes were downregulated. These results suggest that E2F1/E2F2 activity negatively controls growth of mature pancreatic cells and is necessary for the maintenance of differentiated pancreatic phenotypes in the adult.
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NhaA Na+/H+ antiporter mutants that hardly react to the membrane potential.
Directory of Open Access Journals (Sweden)
Dudu Alkoby
Full Text Available pH and Na+ homeostasis in all cells requires Na+/H+ antiporters. The crystal structure, obtained at pH 4, of NhaA, the main antiporter of Escherichia coli, has provided general insights into an antiporter mechanism and its unique pH regulation. Here, we describe a general method to select various NhaA mutants from a library of randomly mutagenized NhaA. The selected mutants, A167P and F267C are described in detail. Both mutants are expressed in Escherichia coli EP432 cells at 70-95% of the wild type but grow on selective medium only at neutral pH, A167P on Li+ (0.1 M and F267C on Na+ (0.6 M. Surprising for an electrogenic secondary transporter, and opposed to wild type NhaA, the rates of A167P and F267C are almost indifferent to membrane potential. Detailed kinetic analysis reveals that in both mutants the rate limiting step of the cation exchange cycle is changed from an electrogenic to an electroneutral reaction.
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Chiu, Din-Chi; Lin, Tzu-Chen; Huang, Wen-I; Cheng, Ting-Jen; Tsai, Keng-Chang; Fang, Jim-Min
2017-11-29
Peramivir is an effective anti-influenza drug in the clinical treatment of influenza, but its efficacy toward the H275Y mutant is reduced. The previously reported cocrystal structures of inhibitors in the mutant neuraminidase (NA) suggest that the hydrophobic side chain should be at the origin of reduced binding affinity. In contrast, zanamivir having a hydrophilic glycerol side chain still possesses high affinity toward the H275Y NA. We thus designed five peramivir analogues (5-9) carrying hydrophilic glycol or glycerol side chains, and evaluated their roles in anti-influenza activity, especially for the H275Y mutant. The synthetic sequence involves a key step of (3 + 2) cycloaddition reactions between alkenes and nitrile oxides to construct the scaffold of peramivir carrying the desired hydrophilic side chains and other appropriate functional groups. The molecular docking experiments reveal that the hydrophilic side chain can provide extra hydrogen bonding with the translocated Glu-276 residue in the H275Y NA active site. Thus, the H275Y mutant may be even more sensitive than wild-type virus toward the peramivir analogues bearing hydrophilic side chains. Notably, the peramivir analogue bearing a glycerol side chain inhibits the H275Y mutant with an IC 50 value of 35 nM, which is better than the WSN virus by 9 fold.
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Cheng, Ning-Hui; Pittman, Jon K; Barkla, Bronwyn J; Shigaki, Toshiro; Hirschi, Kendal D
2003-02-01
The Arabidopsis Ca(2+)/H(+) transporter CAX1 (Cation Exchanger1) may be an important regulator of intracellular Ca(2+) levels. Here, we describe the preliminary localization of CAX1 to the tonoplast and the molecular and biochemical characterization of cax1 mutants. We show that these mutants exhibit a 50% reduction in tonoplast Ca(2+)/H(+) antiport activity, a 40% reduction in tonoplast V-type H(+)-translocating ATPase activity, a 36% increase in tonoplast Ca(2+)-ATPase activity, and increased expression of the putative vacuolar Ca(2+)/H(+) antiporters CAX3 and CAX4. Enhanced growth was displayed by the cax1 lines under Mn(2+) and Mg(2+) stress conditions. The mutants exhibited altered plant development, perturbed hormone sensitivities, and altered expression of an auxin-regulated promoter-reporter gene fusion. We propose that CAX1 regulates myriad plant processes and discuss the observed phenotypes with regard to the compensatory alterations in other transporters.
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Cheng, Ning-Hui; Pittman, Jon K.; Barkla, Bronwyn J.; Shigaki, Toshiro; Hirschi, Kendal D.
2003-01-01
The Arabidopsis Ca2+/H+ transporter CAX1 (Cation Exchanger1) may be an important regulator of intracellular Ca2+ levels. Here, we describe the preliminary localization of CAX1 to the tonoplast and the molecular and biochemical characterization of cax1 mutants. We show that these mutants exhibit a 50% reduction in tonoplast Ca2+/H+ antiport activity, a 40% reduction in tonoplast V-type H+-translocating ATPase activity, a 36% increase in tonoplast Ca2+-ATPase activity, and increased expression of the putative vacuolar Ca2+/H+ antiporters CAX3 and CAX4. Enhanced growth was displayed by the cax1 lines under Mn2+ and Mg2+ stress conditions. The mutants exhibited altered plant development, perturbed hormone sensitivities, and altered expression of an auxin-regulated promoter-reporter gene fusion. We propose that CAX1 regulates myriad plant processes and discuss the observed phenotypes with regard to the compensatory alterations in other transporters. PMID:12566577
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Hicks, Matt N; Gunasekara, Sanjiva; Serate, Jose; Park, Jin; Mosharaf, Pegah; Zhou, Yue; Lee, Jin-Won; Youn, Hwan
2017-10-01
The Escherichia coli cAMP receptor protein (CRP) utilizes the helix-turn-helix motif for DNA binding. The CRP's recognition helix, termed F-helix, includes a stretch of six amino acids (Arg180, Glu181, Thr182, Val183, Gly184, and Arg185) for direct DNA contacts. Arg180, Glu181 and Arg185 are known as important residues for DNA binding and specificity, but little has been studied for the other residues. Here we show that Gly184 is another F-helix residue critical for the transcriptional activation function of CRP. First, glycine was repeatedly selected at CRP position 184 for its unique ability to provide wild type-level transcriptional activation activity. To dissect the glycine requirement, wild type CRP and mutants G184A, G184F, G184S, and G184Y were purified and their in vitro DNA-binding activity was measured. G184A and G184F displayed reduced DNA binding, which may explain their low transcriptional activation activity. However, G184S and G184Y displayed apparently normal DNA affinity. Therefore, an additional factor is needed to account for the diminished transcriptional activation function in G184S and G184Y, and the best explanation is perturbations in their interaction with RNA polymerase. The fact that glycine is the smallest amino acid could not fully warrant its suitability, as shown in this study. We hypothesize that Gly184 fulfills the dual functions of DNA binding and RNA polymerase interaction by conferring conformational flexibility to the F-helix.
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Structure and expression of cytochrome f in an Oenothera plastome mutant.
Johnson, E M; Sears, B B
1990-06-01
The chloroplast mutant pm7 is one of a number of mutants derived from the plastome mutator (pm) line of Oenothera hookeri, strain Johansen. Immunoblotting showed that this mutant accumulates a protein that is cross-antigenic with cytochrome f, but five kilodaltons larger than the mature wild-type protein. Since cytochrome f is known to be translated on plastid ribosomes as a precursor with an amino-terminal extension, it is proposed that the unprocessed cytochrome f precursor accumulates in pm7. In addition to this precursor-sized cytochrome f protein, some mature-sized cytochrome f was also found in the mutant plastids. The pm7 mutation is inherited in a non-Mendelian fashion; but no alterations in chloroplast DNA restriction patterns, or differences in DNA sequence in the region encoding cytochrome f, were found in a comparison of the wild-type and pm7 chloroplast DNAs. Although the mutant was capable of synthesizing heme, no covalently-bound heme, normally found associated with mature, functional, cytochrome f was detected in the mutant at sizes expected for the presumed precursor, or for mature cytochrome f. These results indicate that the aberrant accumulation of a precursor-sized cytochrome f in pm7 is not due to a lesion directly in the plastid gene encoding cytochrome f, petA, or to a deficiency in the ability of the mutant plastids to synthesize or accumulate heme.
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Hua, Ying; Sun, Qi; Wang, Xiangyu; DU, Yanli; Shao, Na; Zhang, Qiwei; Zhao, Wei; Wan, Chengsong
2015-11-01
To construct enterohemorrhagic Escherichia coli (EHEC) O157:H7 strains with delection espF gene and its nucleotide fragment and with espF gene complementation. A pair of homologous arm primers was designed to amplify the gene fragment of kanamycin resistance, which was transformed into EHEC O157:H7 EDL933w strain via the PKD46 plasmid by electroporation. The replacement of the espF gene by kanamycin resistance gene through the PKD46-mediated red recombination system was confirmed by PCR and sequencing. The entire coding region of espF along with its nucleotide fragment was amplified by PCR and cloned into pBAD33 plasmid, which was transformed into a mutant strain to construct the strain with espF complementation. RT-PCR was used to verify the transcription of espF and its nucleotide fragment in the complemented mutant strain. We established EHEC O157:H7 EDL933w strains with espF gene deletion and with espF gene complementation. Both espF and its nucleotide fragment were transcribed in the complemented mutant strain. The two strains provide a basis for further study of the regulatory mechanism of espF.
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Directory of Open Access Journals (Sweden)
Huang He
Full Text Available Chrysanthemum (Chrysanthemum × morifolium is one of the most important ornamental plants in the world. They are typically used as cut flowers or potted plants. Chrysanthemum can exhibit red, purple, pink, yellow and white flowers, but lack bright red and blue flowers. In this study, we identified two chrysanthemum cultivars, C × morifolium 'LPi' and C × morifolium 'LPu', that only accumulate flavonoids in their ligulate flowers. Next, we isolated seven anthocyanin biosynthesis genes, namely CmCHS, CmF3H, CmF3'H, CmDFR, CmANS, CmCHI and Cm3GT in these cultivars. RT-PCR and qRT-PCR analyses showed that CmF3'H was the most important enzyme required for cyanidin biosynthsis. To rebuild the delphinidin pathway, we downregulated CmF3'H using RNAi and overexpressed the Senecio cruentus F3'5'H (PCFH gene in chrysanthemum. The resultant chrysanthemum demonstrated a significantly increased content of cyanidin and brighter red flower petals but did not accumulate delphinidin. These results indicated that CmF3'H in chrysanthemum is important for anthocyanin accumulation, and Senecio cruentus F3'5'H only exhibited F3'H activity in chrysanthemum but did not rebuild the delphinidin pathway to form blue flower chrysanthemum.
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He, Huang; Ke, Hu; Keting, Han; Qiaoyan, Xiang; Silan, Dai
2013-01-01
Chrysanthemum (Chrysanthemum × morifolium) is one of the most important ornamental plants in the world. They are typically used as cut flowers or potted plants. Chrysanthemum can exhibit red, purple, pink, yellow and white flowers, but lack bright red and blue flowers. In this study, we identified two chrysanthemum cultivars, C × morifolium 'LPi' and C × morifolium 'LPu', that only accumulate flavonoids in their ligulate flowers. Next, we isolated seven anthocyanin biosynthesis genes, namely CmCHS, CmF3H, CmF3'H, CmDFR, CmANS, CmCHI and Cm3GT in these cultivars. RT-PCR and qRT-PCR analyses showed that CmF3'H was the most important enzyme required for cyanidin biosynthsis. To rebuild the delphinidin pathway, we downregulated CmF3'H using RNAi and overexpressed the Senecio cruentus F3'5'H (PCFH) gene in chrysanthemum. The resultant chrysanthemum demonstrated a significantly increased content of cyanidin and brighter red flower petals but did not accumulate delphinidin. These results indicated that CmF3'H in chrysanthemum is important for anthocyanin accumulation, and Senecio cruentus F3'5'H only exhibited F3'H activity in chrysanthemum but did not rebuild the delphinidin pathway to form blue flower chrysanthemum.
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Anti-H-Y responses of H-2b mutant mice.
Simpson, E; Gordon, R D; Chandler, P R; Bailey, D
1978-10-01
Two strains of H-2b mutant mice, H-2ba and H-2bf, in which the mutational event took place at H-2K, make anti-H-Y cytotoxic T cell responses which are H-2-restricted, Db-associated and indistinguishable in target cell specificity from those of H-2b mice. Thus, alteration of the H-2K molecule affects neither the Ir gene controlling the response, nor the associative antigen. On the other hand, one H-2Db mutant strain, H-2bo, although it makes a good anti-H-Y cytotoxic response, shows target cell specificity restricted to its own Dbo antigen(s), and neither H-2b, H-2ba or H-2bf anti-H-Y cytotoxic cells kill H-2bo male target cells. Thus, the alteration of the H-2Db molecule does not affect the Ir gene of H-2b mice, but it does alter the H-2Db-associative antigen.
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Phillips, Robert S; Kalu, Ukoha; Hay, Sam
2012-08-21
The effects of pH and hydrostatic pressure on the reaction of H463F tryptophan indole-lyase (TIL) have been evaluated. The mutant TIL shows very low activity for elimination of indole but is still competent to form a quinonoid intermediate from l-tryptophan [Phillips, R. S., Johnson, N., and Kamath, A. V. (2002) Biochemistry 41, 4012-4019]. Stopped-flow measurements show that the formation of the quinonoid intermediate at 505 nm is affected by pH, with a bell-shaped dependence for the forward rate constant, k(f), and dependence on a single basic group for the reverse rate constant, k(r), with the following values: pK(a1) = 8.14 ± 0.15, pK(a2) = 7.54 ± 0.15, k(f,min) = 18.1 ± 1.3 s(-1), k(f,max) = 179 ± 46.3 s(-1), k(r,min) = 11.4 ± 1.2 s(-1), and k(r,max) = 33 ± 1.6 s(-1). The pH effects may be due to ionization of Tyr74 as the base and Cys298 as the acid influencing the rate constant for deprotonation. High-pressure stopped-flow measurements were performed at pH 8, which is the optimum for the forward reaction. The rate constants show an increase with pressure up to 100 MPa and a subsequent decrease above 100 MPa. Fitting the pressure data gives the following values: k(f,0) = 15.4 ± 0.8 s(-1), ΔV(‡) = -29.4 ± 2.9 cm(3) mol(-1), and Δβ(‡) = -0.23 ± 0.03 cm(3) mol(-1) MPa(-1) for the forward reaction, and k(r,0) = 20.7 ± 0.8 s(-1), ΔV(‡) = -9.6 ± 2.3 cm(3) mol(-1), and Δβ(‡) = -0.05 ± 0.02 cm(3) mol(-1) MPa(-1) for the reverse reaction. The primary kinetic isotope effect on quinonoid intermediate formation at pH 8 is small (~2) and is not significantly pressure-dependent, suggesting that the effect of pressure on k(f) may be due to perturbation of an active site preorganization step. The negative activation volume is also consistent with preorganization of the ES complex prior to quinonoid intermediate formation, and the negative compressibility may be due to the effect of pressure on the enzyme conformation. These results support the
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Human liver cell trafficking mutants: characterization and whole exome sequencing.
Directory of Open Access Journals (Sweden)
Fei Yuan
Full Text Available The HuH7 liver cell mutant Trf1 is defective in membrane trafficking and is complemented by the casein kinase 2α subunit CK2α''. Here we identify characteristic morphologies, trafficking and mutational changes in six additional HuH7 mutants Trf2-Trf7. Trf1 cells were previously shown to be severely defective in gap junction functions. Using a Lucifer yellow transfer assay, remarkable attenuation of gap junction communication was revealed in each of the mutants Trf2-Trf7. Electron microscopy and light microscopy of thiamine pyrophosphatase showed that several mutants exhibited fragmented Golgi apparatus cisternae compared to parental HuH7 cells. Intracellular trafficking was investigated using assays of transferrin endocytosis and recycling and VSV G secretion. Surface binding of transferrin was reduced in all six Trf2-Trf7 mutants, which generally correlated with the degree of reduced expression of the transferrin receptor at the cell surface. The mutants displayed the same transferrin influx rates as HuH7, and for efflux rate, only Trf6 differed, having a slower transferrin efflux rate than HuH7. The kinetics of VSV G transport along the exocytic pathway were altered in Trf2 and Trf5 mutants. Genetic changes unique to particular Trf mutants were identified by exome sequencing, and one was investigated in depth. The novel mutation Ile34Phe in the GTPase RAB22A was identified in Trf4. RNA interference knockdown of RAB22A or overexpression of RAB22AI34F in HuH7 cells caused phenotypic changes characteristic of the Trf4 mutant. In addition, the Ile34Phe mutation reduced both guanine nucleotide binding and hydrolysis activities of RAB22A. Thus, the RAB22A Ile34Phe mutation appears to contribute to the Trf4 mutant phenotype.
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Rossi, Raffaella; Granoff, Dan M; Beernink, Peter T
2013-11-04
Factor H-binding protein (fHbp) is a component of a meningococcal vaccine recently licensed in Europe for prevention of serogroup B disease, and a second vaccine in clinical development. The protein specifically binds human factor H (fH), which down-regulates complement activation and enhances resistance to bactericidal activity. There are conflicting data from studies in human fH transgenic mice on whether binding of human fH to fHbp vaccines decreases immunogenicity, and whether mutant fHbp vaccines with decreased fH binding have enhanced immunogenicity. fHbp can be classified into two sub-families based on sequence divergence and immunologic cross-reactivity. Previous studies of mutant fHbp vaccines with low fH binding were from sub-family B, which account for approximately 60% of serogroup B case isolates. In the present study, we evaluated the immunogenicity of two mutant sub-family A fHbp vaccines containing single substitutions, T221A or D211A, which resulted in 15- or 30-fold lower affinity for human fH, respectively, than the corresponding control wild-type fHbp vaccine. In transgenic mice with high serum concentrations of human fH, both mutant vaccines elicited significantly higher IgG titers and higher serum bactericidal antibody responses than the control fHbp vaccine that bound human fH. Thus, mutations introduced into a sub-family A fHbp antigen to decrease fH binding can increase protective antibody responses in human fH transgenic mice. Collectively the data suggest that mutant fHbp antigens with decreased fH binding will result in superior vaccines in humans. Copyright © 2013 Elsevier Ltd. All rights reserved.
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21 CFR 184.1625 - Potassium citrate.
2010-04-01
... acid with potassium hydroxide or potassium carbonate. It occurs as transparent crystals or a white... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Potassium citrate. 184.1625 Section 184.1625 Food... Specific Substances Affirmed as GRAS § 184.1625 Potassium citrate. (a) Potassium citrate (C6H5K3O7·H2O, CAS...
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21 CFR 184.1207 - Calcium lactate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Calcium lactate. 184.1207 Section 184.1207 Food and... Substances Affirmed as GRAS § 184.1207 Calcium lactate. (a) Calcium lactate (C6H10CaO6.xH2O, where x is any... calcium carbonate or calcium hydroxide. (b) The ingredient meets the specifications of the Food Chemicals...
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21 CFR 184.1195 - Calcium citrate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Calcium citrate. 184.1195 Section 184.1195 Food and... Substances Affirmed as GRAS § 184.1195 Calcium citrate. (a) Calcium citrate (Ca3(C6H5O7)2·4H2O, CAS Reg. No. 813-0994-095) is the calcium salt of citric acid. It is prepared by neutralizing citric acid with...
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Dutta, Debodyuti; Mishra, Sabyashachi
2016-11-17
In the fight against bacterial infections and antibiotic resistance, the dapE-encoded N-succinyl-l,l-diaminopimelic acid desuccinylase (DapE) is a potentially safe target enzyme. The role of the Glu134, His67, and His349 residues in the binding and hydrolysis of N-succinyl-l,l-diaminopimelic acid (SDAP) is investigated by employing molecular dynamics simulation and hybrid quantum mechanical-molecular mechanical (MM) calculations of the E134D, H67A, and H349A mutants of DapE. The free energy of substrate binding obtained from the MM-Poisson-Boltzmann surface area approach correctly reproduced the experimentally observed ordering of substrate affinity, that is, E134D > wt > H67A > H349A. The mechanism of catalytic action by the E134D mutant is elucidated by structurally and energetically characterizing the intermediates and the transition states along the reaction pathway. The rate-determining step in the general acid-base hydrolysis reaction by the E134D mutant is found to be the nucleophilic attack step, which involves an activation energy barrier 10 kcal/mol greater than that in the wild-type (wt)-DapE. This explains the 3 orders of magnitude decrease in the experimentally determined k cat value for the E134D mutant compared to that of wt-DapE. In the H67A and H349A mutants, the Glu134 residue undergoes a conformational change and exhibits a strong coordination with the metal centers. This not only results in a weaker substrate binding in the two histidine mutants but also hinders the activation of the catalytic water molecule, which constitutes the first step of the substrate hydrolysis by DapE. This leads to an effective quenching of the catalytic activity in the H67A and H349A mutants.
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Spontaneous and radiation-induced leukemogenesis of the mouse small eye mutant, Pax6Sey3H
International Nuclear Information System (INIS)
Nitta, Yumiko; Satoh, Kenichi; Yoshida, Kazuko; Senba, Kei; Nakagata, Naomi; Peters, J.; Cattanach, B.M.
2004-01-01
Allelic loss on the chromosome 2 is associated with radiation-induced murine acute myeloid leukemia. However, the gene, which contributes mainly to the leukemogenesis has not yet been identified. Expecting any predisposition to acute myeloid leukemia, we performed a radiation leukemogenesis experiment with Pax6 SeY3H , one of the small eye mutants carrying a congenital hemizygosity of the chromosome 2 middle region. A deletion mapping of Pax6 SeY3H with 50 sequence-tagged site (STS) markers indicated that the deleted segment extended between the 106.00 and 111.47 Mb site from the centromere with a length of 5.47 Mb. In the deleted segment, 6 known and 17 novel genes were located. Pax6 SeY3H mutants that crossed back into C3H/He did not develop myeloid leukemia spontaneously, but they did when exposed to gamma-rays. The final incidence of myeloid leukemia in mutants (25.8%) was as high as that in normal sibs (21.4%). Survival curves of leukemia-bearing mutants shifted toward the left (p=0.043 by the Log rank test). F1 hybrids of Pax6 SeY3H with JF1 were less susceptible to radiation than Pax6 SeY3H onto C3H/He in regard to survival (p=0.003 and p<0.00001 for mutants and normal sibs, respectively, by a test of the difference between two proportions). Congenital deletion of the 5.47 Mb segment at the middle region on chromosome 2 alone did not trigger myeloid stem cells to expand clonally in vivo; however, the deletion shortcut the latency of radiation-induced myeloid leukemia. (author)
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2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Riboflavin. 184.1695 Section 184.1695 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1695 Riboflavin. (a) Riboflavin (C17H20N4O6, CAS Reg. No. 83-88-5) occurs...
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21 CFR 184.1443 - Magnesium sulfate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Magnesium sulfate. 184.1443 Section 184.1443 Food... Specific Substances Affirmed as GRAS § 184.1443 Magnesium sulfate. (a) Magnesium sulfate (MgSO4·7H2O, CAS... magnesium oxide, hydroxide, or carbonate with sulfuric acid and evaporating the solution to crystallization...
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21 CFR 184.1446 - Manganese chloride.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Manganese chloride. 184.1446 Section 184.1446 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR... Specific Substances Affirmed as GRAS § 184.1446 Manganese chloride. (a) Manganese chloride (MnCl2·4H2O, CAS...
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21 CFR 184.1426 - Magnesium chloride.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Magnesium chloride. 184.1426 Section 184.1426 Food... Specific Substances Affirmed as GRAS § 184.1426 Magnesium chloride. (a) Magnesium chloride (MgC12·6H2O, CAS... hydrochloric acid solution and crystallizing out magnesium chloride hexahydrate. (b) The ingredient meets the...
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21 CFR 184.1095 - Sulfuric acid.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sulfuric acid. 184.1095 Section 184.1095 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1095 Sulfuric acid. (a) Sulfuric acid (H2SO4, CAS Reg. No. 7664-93-9), also...
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Brewer, John M; Glover, Claiborne V C; Holland, Michael J; Lebioda, Lukasz
2003-05-01
The hypothesis that His159 in yeast enolase moves on a polypeptide loop to protonate the phosphoryl of 2-phosphoglycerate to initiate its conversion to phosphoenolpyruvate was tested by preparing H159N, H159A, and H159F enolases. These have 0.07%-0.25% of the native activity under standard assay conditions and the pH dependence of maximum velocities of H159A and H159N mutants is markedly altered. Activation by Mg2+ is biphasic, with the smaller Mg2+ activation constant closer to that of the "catalytic" Mg2+ binding site of native enolase and the larger in the mM range in which native enolase is inhibited. A third Mg2+ may bind to the phosphoryl, functionally replacing proton donation by His159. N207A enolase lacks an intersubunit interaction that stabilizes the closed loop(s) conformation when 2-phosphoglycerate binds. It has 21% of the native activity, also exhibits biphasic Mg2+ activation, and its reaction with the aldehyde analogue of the substrate is more strongly inhibited than is its normal enzymatic reaction. Polypeptide loop(s) closure may keep a proton from His159 interacting with the substrate phosphoryl oxygen long enough to stabilize a carbanion intermediate.
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Directory of Open Access Journals (Sweden)
Peter T Beernink
Full Text Available The binding of human complement inhibitors to vaccine antigens in vivo could diminish their immunogenicity. A meningococcal ligand for the complement down-regulator, factor H (fH, is fH-binding protein (fHbp, which is specific for human fH. Vaccines containing recombinant fHbp or native outer membrane vesicles (NOMV from mutant strains with over-expressed fHbp are in clinical development. In a previous study in transgenic mice, the presence of human fH impaired the immunogenicity of a recombinant fHbp vaccine. In the present study, we prepared two NOMV vaccines from mutant group B strains with over-expressed wild-type fHbp or an R41S mutant fHbp with no detectable fH binding. In wild-type mice in which mouse fH did not bind to fHbp in either vaccine, the NOMV vaccine with wild-type fHbp elicited 2-fold higher serum IgG anti-fHbp titers (P = 0.001 and 4-fold higher complement-mediated bactericidal titers against a PorA-heterologous strain than the NOMV with the mutant fHbp (P = 0.003. By adsorption, the bactericidal antibodies were shown to be directed at fHbp. In transgenic mice in which human fH bound to the wild-type fHbp but not to the R41S fHbp, the NOMV vaccine with the mutant fHbp elicited 5-fold higher serum IgG anti-fHbp titers (P = 0.002, and 19-fold higher bactericidal titers than the NOMV vaccine with wild-type fHbp (P = 0.001. Thus, in mice that differed only by the presence of human fH, the respective results with the two vaccines were opposite. The enhanced bactericidal activity elicited by the mutant fHbp vaccine in the presence of human fH far outweighed the loss of immunogenicity of the mutant protein in wild-type animals. Engineering fHbp not to bind to its cognate complement inhibitor, therefore, may increase vaccine immunogenicity in humans.
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Energy Technology Data Exchange (ETDEWEB)
Likar, Yury; Dobrenkov, Konstantin; Olszewska, Malgorzata; Shenker, Larissa; Hricak, Hedvig; Ponomarev, Vladimir [Memorial Sloan-Kettering Cancer Center, Molecular Imaging Laboratory, Department of Radiology, New York, NY (United States); Cai, Shangde [Memorial Sloan-Kettering Cancer Center, Radiochemistry/Cyclotron Core Facility, New York, NY (United States)
2009-08-15
The aim of this study was to create an alternative mutant of the herpes simplex virus type 1 thymidine kinase (HSV1-tk) reporter gene with reduced phosphorylation capacity for acycloguanosine derivatives, but not pyrimidine-based compounds that will allow for successful PET imaging. A new mutant of HSV1-tk reporter gene, suitable for PET imaging using pyrimidine-based radiotracers, was developed. The HSV1-tk mutant contains an arginine-to-glutamine substitution at position 176 (HSV1-R176Qtk) of the nucleoside binding region of the enzyme. The mutant-gene product showed favorable enzymatic characteristics toward pyrimidine-based nucleosides, while exhibiting reduced activity with acycloguanosine derivatives. In order to enhance HSV1-R176Qtk reporter activity with pyrimidine-based radiotracers, we introduced the R176Q substitution into the more active HSV1-sr39tk mutant. U87 human glioma cells transduced with the HSV1-R176Qsr39tk double mutant reporter gene showed high {sup 3}H-FEAU pyrimidine nucleoside and low {sup 3}H-penciclovir acycloguanosine analog uptake in vitro. PET imaging also demonstrated high {sup 18}F-FEAU and low {sup 18}F-FHBG accumulation in HSV1-R176Qsr39tk+ xenografts. The feasibility of imaging two independent nucleoside-specific HSV1-tk mutants in the same animal with PET was demonstrated. Two opposite xenografts expressing the HSV1-R176Qsr39tk reporter gene and the previously described acycloguanosine-specific mutant of HSV1-tk, HSV1-A167Ysr39tk reporter gene, were imaged using a short-lived pyrimidine-based {sup 18}F-FEAU and an acycloguanosine-based {sup 18}F-FHBG radiotracer, respectively, administered on 2 consecutive days. We conclude that in combination with acycloguanosine-specific HSV1-A167Ysr39tk reporter gene, a HSV1-tk mutant containing the R176Q substitution could be used for PET imaging of two different cell populations or concurrent molecular biological processes in the same living subject. (orig.)
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Directory of Open Access Journals (Sweden)
Sujit Roy
Full Text Available Abscisic acid (ABA acts as an important plant hormone in regulating various aspects of plant growth and developmental processes particularly under abiotic stress conditions. An increased ABA level in plant cells inhibits DNA replication and cell division, causing plant growth retardation. In this study, we have investigated the effects of ABA on the growth responses of some major loss-of-function mutants of DNA double-stand break (DSB repair genes in Arabidopsis during seed germination and early stages of seedling growth for understanding the role of ABA in the induction of genome instability in plants. A comparative analysis of ABA sensitivity of wild-type Arabidopsis and the knockout mutant lines related to DSB sensors, including atatm, atatr, the non-homologous end joining (NHEJ pathway genes, and mutants related to homologous recombination (HR pathway genes showed relatively enhanced sensitivity of atatr and HR-related mutants to ABA treatment. The expression levels of HR-related genes were increased in wild-type Arabidopsis (Col-0 during seed germination and early stages of seedling growth. Immunoblotting experiments detected phosphorylation of histone H2AX in wild-type (Col-0 and DSB repair gene mutants after ABA treatment, indicating the activation of DNA damage response due to ABA treatment. Analyses of DSB repair kinetics using comet assay under neutral condition have revealed comparatively slower DSB repair activity in HR mutants. Overall, our results have provided comprehensive information on the possible effect of ABA on DNA repair machinery in plants and also indicated potential functional involvement of HR pathway in repairing ABA induced DNA damage in Arabidopsis.
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21 CFR 184.1229 - Calcium stearate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Calcium stearate. 184.1229 Section 184.1229 Food... Specific Substances Affirmed as GRAS § 184.1229 Calcium stearate. (a) Calcium stearate (Ca(C17H35COO)2, CAS Reg. No. 1529-23-0) is the calcium salt of stearic acid derived from edible sources. It is prepared as...
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21 CFR 184.1206 - Calcium iodate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Calcium iodate. 184.1206 Section 184.1206 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1206 Calcium iodate. (a) Calcium iodate [Ca(IO3)2·H2O, CAS Reg. No. 7789-80...
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21 CFR 184.1193 - Calcium chloride.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Calcium chloride. 184.1193 Section 184.1193 Food... Specific Substances Affirmed as GRAS § 184.1193 Calcium chloride. (a) Calcium chloride (CaCl2·2H2O, CAS Reg. No. 10035-04-8) or anhydrous calcium chloride (CaCl2, CAS Reg. No. 10043-52-4) may be commercially...
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21 CFR 184.1061 - Lactic acid.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Lactic acid. 184.1061 Section 184.1061 Food and... Substances Affirmed as GRAS § 184.1061 Lactic acid. (a) Lactic acid (C3H6O3, CAS Reg. Nos.: dl mixture, 598... hydrogen cyanide and subsequent hydrolysis to lactic acid. (b) The ingredient meets the specifications of...
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21 CFR 184.1081 - Propionic acid.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Propionic acid. 184.1081 Section 184.1081 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1081 Propionic acid. (a) Propionic acid (C3H6O2, CAS Reg. No. 79-09-4) is...
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21 CFR 184.1021 - Benzoic acid.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Benzoic acid. 184.1021 Section 184.1021 Food and... Substances Affirmed as GRAS § 184.1021 Benzoic acid. (a) Benzoic acid is the chemical benzenecarboxylic acid (C7H6O2), occurring in nature in free and combined forms. Among the foods in which benzoic acid occurs...
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21 CFR 184.1065 - Linoleic acid.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Linoleic acid. 184.1065 Section 184.1065 Food and... Substances Affirmed as GRAS § 184.1065 Linoleic acid. (a) Linoleic acid ((Z, Z)-9, 12-octadecadienoic acid (C17H31COOH) (CAS Reg. No. 60-33-3)), a straight chain unsaturated fatty acid with a molecular weight of 280.5...
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21 CFR 184.1091 - Succinic acid.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Succinic acid. 184.1091 Section 184.1091 Food and... Substances Affirmed as GRAS § 184.1091 Succinic acid. (a) Succinic acid (C4H6O4, CAS Reg. No. 110-15-6), also referred to as amber acid and ethylenesuccinic acid, is the chemical 1,4-butanedioic acid. It is...
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21 CFR 184.1090 - Stearic acid.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Stearic acid. 184.1090 Section 184.1090 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1090 Stearic acid. (a) Stearic acid (C18H36O2, CAS Reg. No. 57-11-4) is a...
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21 CFR 184.1261 - Copper sulfate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Copper sulfate. 184.1261 Section 184.1261 Food and... Substances Affirmed as GRAS § 184.1261 Copper sulfate. (a) Copper sulfate (cupric sulfate, CuSO4·5H2O, CAS... the reaction of sulfuric acid with cupric oxide or with copper metal. (b) The ingredient must be of a...
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Microstructure and Hardness Variation in a TIG Weldment of Irradiated F82H
International Nuclear Information System (INIS)
Tanigawa, H.; Ando, M.; Sawai, T.; Shiba, K.; Hashimoto, N.; Klueh, R.L.
2003-01-01
Previous work reported that a TIG weld joint of F82H exhibited low irradiation hardening in a tensile test, compared to the base metal. Microhardness tests and microstructure observation on the neutron-irradiated TIG weld joint of F82H revealed that the over-tempered zone in the heat-affected zone (HAZ) exhibited this good performance. The region in the HAZ where the prior austenite grain size became very fine during welding also exhibited lower irradiation hardening. Hypotheses for these low-hardening mechanisms were proposed based on the phase diagram and grain size
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21 CFR 184.1185 - Calcium acetate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Calcium acetate. 184.1185 Section 184.1185 Food and... Substances Affirmed as GRAS § 184.1185 Calcium acetate. (a) Calcium acetate (Ca (C2H3O2)2, CAS Reg. No. 62-54-4), also known as acetate of lime or vinegar salts, is the calcium salt of acetic acid. It may be...
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2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Corn sugar. 184.1857 Section 184.1857 Food and... Substances Affirmed as GRAS § 184.1857 Corn sugar. (a) Corn sugar (C6H12O6, CAS Reg. No. 50-99-7), commonly... monohydrate form and is produced by the complete hydrolysis of corn starch with safe and suitable acids or...
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2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Vitamin D. 184.1950 Section 184.1950 Food and Drugs... Substances Affirmed as GRAS § 184.1950 Vitamin D. (a) Vitamin D is added to food as the following food ingredients: (1) Crystalline vitamin D2 (C28H44O, CAS Reg. No. 50-14-6), also known as ergocalciferol, is the...
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21 CFR 184.1005 - Acetic acid.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Acetic acid. 184.1005 Section 184.1005 Food and... Substances Affirmed as GRAS § 184.1005 Acetic acid. (a) Acetic acid (C2H4O2, CAS Reg. No. 64-19-7) is known as ethanoic acid. It occurs naturally in plant and animal tissues. It is produced by fermentation of...
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2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Malic acid. 184.1069 Section 184.1069 Food and... Substances Affirmed as GRAS § 184.1069 Malic acid. (a) Malic acid (C4H6O5, CAS Reg. No. of L-form 97-67-6, CAS Reg. No. of DL-form 617-48-1) is the common name for 1-hydroxy-1, 2-ethanedicarboxylic acid. L...
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Pleiotropic effects of hemagglutinin amino acid substitutions of H5 influenza escape mutants
International Nuclear Information System (INIS)
Rudneva, Irina A.; Timofeeva, Tatiana A.; Ignatieva, Anna V.; Shilov, Aleksandr A.; Krylov, Petr S.; Ilyushina, Natalia A.; Kaverin, Nikolai V.
2013-01-01
In the present study we assessed pleiotropic characteristics of the antibody-selected mutations. We examined pH optimum of fusion, temperatures of HA heat inactivation, and in vitro and in vivo replication kinetics of the previously obtained influenza H5 escape mutants. Our results showed that HA1 N142K mutation significantly lowered the pH of fusion optimum. Mutations of the escape mutants located in the HA lateral loop significantly affected H5 HA thermostability (P<0.05). HA changes at positions 131, 144, 145, and 156 and substitutions at positions 131, 142, 145, and 156 affected the replicative ability of H5 escape mutants in vitro and in vivo, respectively. Overall, a co-variation between antigenic specificity and different HA phenotypic properties has been demonstrated. We believe that the monitoring of pleiotropic effects of the HA mutations found in H5 escape mutants is essential for accurate prediction of mutants with pandemic potential. - Highlights: • HA1 N142K mutation significantly lowered the pH of fusion optimum. • Mutations located in the HA lateral loop significantly affected H5 HA thermostability. • HA changes at positions 131, 142, 144, 145, and 156 affected the replicative ability of H5 mutants. • Acquisition of glycosylation site could lead to the emergence of multiple pleiotropic effects
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Pleiotropic effects of hemagglutinin amino acid substitutions of H5 influenza escape mutants
Energy Technology Data Exchange (ETDEWEB)
Rudneva, Irina A.; Timofeeva, Tatiana A.; Ignatieva, Anna V.; Shilov, Aleksandr A.; Krylov, Petr S. [D.I. Ivanovsky Institute of Virology, 123098 Moscow (Russian Federation); Ilyushina, Natalia A., E-mail: Natalia.Ilyushina@fda.hhs.gov [FDA CDER, 29 Lincoln Drive, Bethesda, MD 20892 (United States); Kaverin, Nikolai V., E-mail: nik.kaverin@gmail.com [D.I. Ivanovsky Institute of Virology, 123098 Moscow (Russian Federation)
2013-12-15
In the present study we assessed pleiotropic characteristics of the antibody-selected mutations. We examined pH optimum of fusion, temperatures of HA heat inactivation, and in vitro and in vivo replication kinetics of the previously obtained influenza H5 escape mutants. Our results showed that HA1 N142K mutation significantly lowered the pH of fusion optimum. Mutations of the escape mutants located in the HA lateral loop significantly affected H5 HA thermostability (P<0.05). HA changes at positions 131, 144, 145, and 156 and substitutions at positions 131, 142, 145, and 156 affected the replicative ability of H5 escape mutants in vitro and in vivo, respectively. Overall, a co-variation between antigenic specificity and different HA phenotypic properties has been demonstrated. We believe that the monitoring of pleiotropic effects of the HA mutations found in H5 escape mutants is essential for accurate prediction of mutants with pandemic potential. - Highlights: • HA1 N142K mutation significantly lowered the pH of fusion optimum. • Mutations located in the HA lateral loop significantly affected H5 HA thermostability. • HA changes at positions 131, 142, 144, 145, and 156 affected the replicative ability of H5 mutants. • Acquisition of glycosylation site could lead to the emergence of multiple pleiotropic effects.
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Wang, T H; Wang, S Y; Wang, X D; Jiang, H Q; Yang, Y Q; Wang, Y; Cheng, J L; Zhang, C T; Liang, W W; Feng, H L
2018-05-21
Oxidative stress exhibits a central role in the course of amyotrophic lateral sclerosis (ALS), a progressive neurodegenerative disease commonly found to include a copper/zinc superoxide dismutase (SOD1) gene mutation. Fisetin, a natural antioxidant, has shown benefits in varied neurodegenerative diseases. The possible effect of fisetin in ALS has not been clarified as of yet. We investigated whether fisetin affected mutant hSOD1 ALS models. Three different hSOD1-related mutant models were used: Drosophila expressing mutant hSOD1 G85R , hSOD1 G93A NSC34 cells, and transgenic mice. Fisetin treatment provided neuroprotection as demonstrated by an improved survival rate, attenuated motor impairment, reduced ROS damage and regulated redox homeostasis compared with those in controls. Furthermore, fisetin increased the expression of phosphorylated ERK and upregulated antioxidant factors, which were reversed by MEK/ERK inhibition. Finally, fisetin reduced the levels of both mutant and wild-type hSOD1 in vivo and in vitro, as well as the levels of detergent-insoluble hSOD1 proteins. The results indicate that fisetin protects cells from ROS damage and improves the pathological behaviors caused by oxidative stress in disease models related to SOD1 gene mutations probably by activating ERK, thereby providing a potential treatment for ALS. Copyright © 2018 IBRO. Published by Elsevier Ltd. All rights reserved.
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Cellular androgen content influences enzalutamide agonism of F877L mutant androgen receptor
Coleman, Daniel J.; Van Hook, Kathryn; King, Carly J.; Schwartzman, Jacob; Lisac, Robert; Urrutia, Joshua; Sehrawat, Archana; Woodward, Josha; Wang, Nicholas J.; Gulati, Roman; Thomas, George V.; Beer, Tomasz M.; Gleave, Martin; Korkola, James E.; Gao, Lina; Heiser, Laura M.; Alumkal, Joshi J.
2016-01-01
Prostate cancer is the most commonly diagnosed and second-most lethal cancer among men in the United States. The vast majority of prostate cancer deaths are due to castration-resistant prostate cancer (CRPC) – the lethal form of the disease that has progressed despite therapies that interfere with activation of androgen receptor (AR) signaling. One emergent resistance mechanism to medical castration is synthesis of intratumoral androgens that activate the AR. This insight led to the development of the AR antagonist enzalutamide. However, resistance to enzalutamide invariably develops, and disease progression is nearly universal. One mechanism of resistance to enzalutamide is an F877L mutation in the AR ligand-binding domain that can convert enzalutamide to an agonist of AR activity. However, mechanisms that contribute to the agonist switch had not been fully clarified, and there were no therapies to block AR F877L. Using cell line models of castration-resistant prostate cancer (CRPC), we determined that cellular androgen content influences enzalutamide agonism of mutant F877L AR. Further, enzalutamide treatment of AR F877L-expressing cell lines recapitulated the effects of androgen activation of F877L AR or wild-type AR. Because the BET bromodomain inhibitor JQ-1 was previously shown to block androgen activation of wild-type AR, we tested JQ-1 in AR F877L-expressing CRPC models. We determined that JQ-1 suppressed androgen or enzalutamide activation of mutant F877L AR and suppressed growth of mutant F877L AR CRPC tumors in vivo, demonstrating a new strategy to treat tumors harboring this mutation. PMID:27276681
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Energy Technology Data Exchange (ETDEWEB)
Leban, I. (Dept. of Chemistry and Chemical Technology, Univ. Ljubljana (Slovenia))
1994-06-15
The crystals exhibit racemic twinning. The structure consists of hydrazinium(1+), N[sub 2]H[sub 5][sup +], cations and usual octahedral hexafluorotitanate(IV) anions. They are linked together via hydrogen bonds of the types N-H..F and N-H..N. (orig.).
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21 CFR 184.1945 - Vitamin B12.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Vitamin B12. 184.1945 Section 184.1945 Food and... Substances Affirmed as GRAS § 184.1945 Vitamin B12. (a) Vitamin B12, also known as cyanocobalamin (C63H88Co... is used in food at levels not to exceed current good manufacturing practice. Vitamin B12 also may be...
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Hydrazinium(1+) hexafluorotitanate(IV), 2N2H5+.TiF62-
International Nuclear Information System (INIS)
Leban, I.
1994-01-01
The crystals exhibit racemic twinning. The structure consists of hydrazinium(1+), N 2 H 5 + , cations and usual octahedral hexafluorotitanate(IV) anions. They are linked together via hydrogen bonds of the types N-H..F and N-H..N. (orig.)
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RpfF-dependent regulon of Xylella fastidiosa.
Wang, Nian; Li, Jian-Liang; Lindow, Steven E
2012-11-01
ABSTRACT Xylella fastidiosa regulates traits important to both virulence of grape as well as colonization of sharpshooter vectors via its production of a fatty acid signal molecule known as DSF whose production is dependent on rpfF. Although X. fastidiosa rpfF mutants exhibit increased virulence to plants, they are unable to be spread from plant to plant by insect vectors. To gain more insight into the traits that contribute to these processes, a whole-genome Agilent DNA microarray for this species was developed and used to determine the RpfF-dependent regulon by transcriptional profiling. In total, 446 protein coding genes whose expression was significantly different between the wild type and an rpfF mutant (false discovery rate genes were downregulated in the rpfF mutant compared with the wild-type strain whereas 281 genes were over-expressed. RpfF function was required for regulation of 11 regulatory and σ factors, including rpfE, yybA, PD1177, glnB, rpfG, PD0954, PD0199, PD2050, colR, rpoH, and rpoD. In general, RpfF is required for regulation of genes involved in attachment and biofilm formation, enhancing expression of hemagglutinin genes hxfA and hxfB, and suppressing most type IV pili and gum genes. A large number of other RpfF-dependent genes that might contribute to virulence or insect colonization were also identified such as those encoding hemolysin and colicin V, as well as genes with unknown functions.
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International Nuclear Information System (INIS)
Touvin, H.
1973-01-01
An early homogeneous mutant line (B) was obtained in M 2 generation following the gamma-rays (15kR) treatment of the dry seeds of a fixed homogeneous line of soft wheat (A). The study of this mutant leads to the following observations: the earliness is stable in the mutant stock during successive generations and in different climatic conditions; the products of reciprocal crosses between the mutant B and the original line A, compared in micro-tests under greenhouse conditions gave following different results according to the cross; in the F 1 , the reciprocal hybrids (AxB, BxA) are earlier than the mutant. The F 1 offsprings (BxA) which received the earliness characteristic from the female parent, develop more rapidly than the reciprocal F 1 hybrid (AxB). Thus, this shows that there exist a maternal effect from the mutant side. In the F 2 , the descendants of the hybrid (AxB) segregated phenotypically in two classes, early and late, whereas the other hybrid (BxA) produces only early plants. The F 3 offsprings confirm the observations made in the F 2 generation. Although the F 3 generation of the hybrid (AxB) is composed of the early homogenous, the heterogeneous and the late homogeneous lines, but no segregation occurs in the cross (BxA). The segregation ratio in F 2 and in most of the backcrosses progenies indicates that the transmission of the earliness character is monogenic. From these results the existence of a major gene for earliness can be supposed, the expression of which appears to be under the control of the cytoplasm. The conclusion emphasizes the importance of the reciprocal crosses in the use of the mutants [fr
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International Nuclear Information System (INIS)
Landry, L.G.; Last, R.L.; Chapple, C.C.S.
1995-01-01
We have assessed ultraviolet-B (UV-B)-induced injury in wild-type Arabidopsis thaliana and two mutants with altered aromatic secondary product biosynthesis. Arabidopsis mutants defective in the ability to synthesize UV-B-absorbing compounds (flavonoids in transparent testa 5 [tt5] and sinapate esters in ferulic acid hydroxylase 1 [fah 1]) are more sensitive to UV-B than is the wild-type Landsberg erecta. Despite its ability to accumulate UV-absorptive flavonoid compounds, the ferulic acid hydroxylase mutant fah1 exhibits more physiological injury (growth inhibition and foliar lesions) than either wild type or tt5. The extreme UV-B sensitivity of fah1 demonstrates the importance of hydroxycinnamate esters as UV-B protectants. Consistent with the whole-plant response, the highest levels of lipid and protein oxidation products were seen in fah1. Ascorbate peroxidase enzyme activity was also increased in the leaves of UV-B-treated plants in a dose- and genotype-dependent manner. These results demonstrate that, in A. thaliana, hydryoxycinnamates are more effective UV-B protectants than flavonoids. The data also indicate that A. thaliana responds to UV-B as an oxidative stress, and sunscreen compounds reduce the oxidative damage caused by UV-B. 36 refs., 6 figs
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Selenolate complexes of CYP101 and the heme-bound hHO-1/H25A proximal cavity mutant.
Jiang, Yongying; Ortiz de Montellano, Paul R
2008-05-05
Thiolate and selenolate complexes of CYP101 (P450cam) and the H25A proximal cavity mutant of heme-bound human heme oxygenase-1 (hHO-1) have been examined by UV-vis spectroscopy. Both thiolate and selenolate ligands bound to the heme distal side in CYP101 and gave rise to characteristic hyperporphyrin spectra. Thiolate ligands also bound to the proximal side of the heme in the cavity created by the H25A mutation in hHO-1, giving a Soret absorption similar to that of the H25C hHO-1 mutant. Selenolate ligands also bound to this cavity mutant under anaerobic conditions but reduced the heme iron to the ferrous state, as shown by the formation of a ferrous CO complex. Under aerobic conditions, the selenolate ligand but not the thiolate ligand was rapidly oxidized. These results indicate that selenocysteine-coordinated heme proteins will not be stable species in the absence of a redox potential stabilizing effect.
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Directory of Open Access Journals (Sweden)
Tshidi Tsibane
Full Text Available Most monoclonal antibodies (mAbs to the influenza A virus hemagglutinin (HA head domain exhibit very limited breadth of inhibitory activity due to antigenic drift in field strains. However, mAb 1F1, isolated from a 1918 influenza pandemic survivor, inhibits select human H1 viruses (1918, 1943, 1947, and 1977 isolates. The crystal structure of 1F1 in complex with the 1918 HA shows that 1F1 contacts residues that are classically defined as belonging to three distinct antigenic sites, Sa, Sb and Ca(2. The 1F1 heavy chain also reaches into the receptor binding site (RBS and interacts with residues that contact sialoglycan receptors and determine HA receptor specificity. The 1F1 epitope is remarkably similar to the previously described murine HC63 H3 epitope, despite significant sequence differences between H1 and H3 HAs. Both antibodies potently inhibit receptor binding, but only HC63 can block the pH-induced conformational changes in HA that drive membrane fusion. Contacts within the RBS suggested that 1F1 may be sensitive to changes that alter HA receptor binding activity. Affinity assays confirmed that sequence changes that switch the HA to avian receptor specificity affect binding of 1F1 and a mAb possessing a closely related heavy chain, 1I20. To characterize 1F1 cross-reactivity, additional escape mutant selection and site-directed mutagenesis were performed. Residues 190 and 227 in the 1F1 epitope were found to be critical for 1F1 reactivity towards 1918, 1943 and 1977 HAs, as well as for 1I20 reactivity towards the 1918 HA. Therefore, 1F1 heavy-chain interactions with conserved RBS residues likely contribute to its ability to inhibit divergent HAs.
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Baldwin, Thomas T; Basenko, Evelina; Harb, Omar; Brown, Neil A; Urban, Martin; Hammond-Kosack, Kim E; Bregitzer, Phil P
2018-06-01
There is no comprehensive storage for generated mutants of Fusarium graminearum or data associated with these mutants. Instead, researchers relied on several independent and non-integrated databases. FgMutantDb was designed as a simple spreadsheet that is accessible globally on the web that will function as a centralized source of information on F. graminearum mutants. FgMutantDb aids in the maintenance and sharing of mutants within a research community. It will serve also as a platform for disseminating prepublication results as well as negative results that often go unreported. Additionally, the highly curated information on mutants in FgMutantDb will be shared with other databases (FungiDB, Ensembl, PhytoPath, and PHI-base) through updating reports. Here we describe the creation and potential usefulness of FgMutantDb to the F. graminearum research community, and provide a tutorial on its use. This type of database could be easily emulated for other fungal species. Published by Elsevier Inc.
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Tolerance of photoperiod insensitive mutant of Sesbania rostrata to salinity and pH
International Nuclear Information System (INIS)
Ramani, Saradha; Joshua, D.C.; Shaikh, M.S.; Athalye, V.V.
1998-01-01
The photoperiod insensitive mutant, TSR-1 of Sesbania rostrata was compared with the parent variety for its response to soil salinity and different levels of pH in hydroponics. The plant growth and stem nodulation were not significantly affected by salinity. However, salinity in soil without farmyard manure stimulated plant growth. Radiotracer studies showed that the translocation of Na to stem and leaves was much less compared to uptake in both parent and mutant. The growth of TSR-1 was comparable to or marginally better than that of the parent variety in the pH range of 3.5-8.0. Root nodulation was less with low pH. The nitrogen content was not adversely affected by pH, but it was reduced with 200 mM NaCl. This mutant in addition to being short-day insensitive, is tolerant to low to moderate salinity levels and pH like its parent. (author)
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International Nuclear Information System (INIS)
Chen Pingping; Zhang Xiaoming; Ye Shenghai; Zhao Ningchun; Lu Yanting; Liu Heqin; Jin Qingsheng; Yang Ling
2010-01-01
A lesion mimic mutant,obtained by radiation mutagenesis on the seeds of a japonica rice variety Zhejing 22, exhibited a lesion mimic phenotype during the whole growth stage under different environments. Genetic analysis indicated that the mutant trait was controlled by a single recessive gene named spl (t). Relying on simple sequence repeat (SSR) and recessive class analysis method to map the spl (t) gene with a F 2 population was constructed by crossing the mutant spl (t) with Zhenshan 97B.spl (t) was mapped in the interval of 0.8cM between RM7195 and RM27929 near centromere region on the short arm of chromosome 12.Blue trypan dye analyses indicated that the lesion mimic trait of the mutant was caused by the programmer cell death. Further study showed that the programmer cell death was caused by H 2 O 2 oxidative burst. By inoculation of bacterial leaf blight and blast strains, the resistances of the mutant were similar to the wild variety Zhejing 22. (authors)
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Loss-of-function and gain-of-function phenotypes of stomatocytosis mutant RhAG F65S
Stewart, Andrew K.; Shmukler, Boris E.; Vandorpe, David H.; Rivera, Alicia; Heneghan, John F.; Li, Xiaojin; Hsu, Ann; Karpatkin, Margaret; O'Neill, Allison F.; Bauer, Daniel E.; Heeney, Matthew M.; John, Kathryn; Kuypers, Frans A.; Gallagher, Patrick G.; Lux, Samuel E.; Brugnara, Carlo; Westhoff, Connie M.
2011-01-01
Four patients with overhydrated cation leak stomatocytosis (OHSt) exhibited the heterozygous RhAG missense mutation F65S. OHSt erythrocytes were osmotically fragile, with elevated Na and decreased K contents and increased cation channel-like activity. Xenopus oocytes expressing wild-type RhAG and RhAG F65S exhibited increased ouabain and bumetanide-resistant uptake of Li+ and 86Rb+, with secondarily increased 86Rb+ influx sensitive to ouabain and to bumetanide. Increased RhAG-associated 14C-methylammonium (MA) influx was severely reduced in RhAG F65S-expressing oocytes. RhAG-associated influxes of Li+, 86Rb+, and 14C-MA were pharmacologically distinct, and Li+ uptakes associated with RhAG and RhAG F65S were differentially inhibited by NH4+ and Gd3+. RhAG-expressing oocytes were acidified and depolarized by 5 mM bath NH3/NH4+, but alkalinized and depolarized by subsequent bath exposure to 5 mM methylammonium chloride (MA/MA+). RhAG F65S-expressing oocytes exhibited near-wild-type responses to NH4Cl, but MA/MA+ elicited attenuated alkalinization and strong hyperpolarization. Expression of RhAG or RhAG F65S increased steady-state cation currents unaltered by bath Li+ substitution or bath addition of 5 mM NH4Cl or MA/MA+. These oocyte studies suggest that 1) RhAG expression increases oocyte transport of NH3/NH4+ and MA/MA+; 2) RhAG F65S exhibits gain-of-function phenotypes of increased cation conductance/permeability, and loss-of-function phenotypes of decreased and modified MA/MA+ transport, and decreased NH3/NH4+-associated depolarization; and 3) RhAG transports NH3/NH4+ and MA/MA+ by distinct mechanisms, and/or the substrates elicit distinct cellular responses. Thus, RhAG F65S is a loss-of-function mutation for amine transport. The altered oocyte intracellular pH, membrane potential, and currents associated with RhAG or RhAG F65S expression may reflect distinct transport mechanisms. PMID:21849667
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21 CFR 184.1697 - Riboflavin-5′-phosphate (sodium).
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Riboflavin-5â²-phosphate (sodium). 184.1697 Section... SAFE Listing of Specific Substances Affirmed as GRAS § 184.1697 Riboflavin-5′-phosphate (sodium). (a) Riboflavin-5′-phosphate (sodium) (C17H20N4O9PNa·2H2O, CAS Reg. No 130-40-5) occurs as the dihydrate in yellow...
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Over-expression of p53 mutants in LNCaP cells alters tumor growth and angiogenesis in vivo
International Nuclear Information System (INIS)
Perryman, L.A.; Blair, J.M.; Kingsley, E.A.; Szymanska, B.; Ow, K.T.; Wen, V.W.; MacKenzie, K.L.; Vermeulen, P.B.; Jackson, P.; Russell, P.J.
2006-01-01
This study has investigated the impact of three specific dominant-negative p53 mutants (F134L, M237L, and R273H) on tumorigenesis by LNCaP prostate cancer cells. Mutant p53 proteins were associated with an increased subcutaneous 'take rate' in NOD-SCID mice, and increased production of PSA. Tumors expressing F134L and R273H grew slower than controls, and were associated with decreased necrosis and apoptosis, but not hypoxia. Interestingly, hypoxia levels were increased in tumors expressing M237L. There was less proliferation in F134L-bearing tumors compared to control, but this was not statistically significant. Angiogenesis was decreased in tumors expressing F134L and R273H compared with M237L, or controls. Conditioned medium from F134L tumors inhibited growth of normal human umbilical-vein endothelial cells but not telomerase-immortalized bone marrow endothelial cells. F134L tumor supernatants showed lower levels of VEGF and endostatin compared with supernatants from tumors expressing other mutants. Our results support the possibility that decreased angiogenesis might account for reduced growth rate of tumor cells expressing the F134L p53 mutation
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Bele, Aditya; Mirza, Sameer; Zhang, Ying; Ahmad Mir, Riyaz; Lin, Simon; Kim, Jun Hyun; Gurumurthy, Channabasavaiah Basavaraju; West, William; Qiu, Fang; Band, Hamid; Band, Vimla
2015-01-01
The mammalian ortholog of Drosophila ecdysoneless (Ecd) gene product regulates Rb-E2F interaction and is required for cell cycle progression. Ecd is overexpressed in breast cancer and its overexpression predicts shorter survival in patients with ErbB2-positive tumors. Here, we demonstrate Ecd knock down (KD) in human mammary epithelial cells (hMECs) induces growth arrest, similar to the impact of Ecd Knock out (KO) in mouse embryonic fibroblasts. Furthermore, whole-genome mRNA expression analysis of control vs. Ecd KD in hMECs demonstrated that several of the top 40 genes that were down-regulated were E2F target genes. To address the role of Ecd in mammary oncogenesis, we overexpressed Ecd and/or mutant H-Ras in hTERT-immortalized hMECs. Cell cycle analyses revealed hMECs overexpressing Ecd+Ras showed incomplete arrest in G1 phase upon growth factor deprivation, and more rapid cell cycle progression in growth factor-containing medium. Analyses of cell migration, invasion, acinar structures in 3-D Matrigel and anchorage-independent growth demonstrated that Ecd+Ras-overexpressing cells exhibit substantially more dramatic transformed phenotype as compared to cells expressing vector, Ras or Ecd. Under conditions of nutrient deprivation, Ecd+Ras-overexpressing hMECs exhibited better survival, with substantial upregulation of the autophagy marker LC3 both at the mRNA and protein levels. Significantly, while hMECs expressing Ecd or mutant Ras alone did not form tumors in NOD/SCID mice, Ecd+Ras-overexpressing hMECs formed tumors, clearly demonstrating oncogenic cooperation between Ecd and mutant Ras. Collectively, we demonstrate an important co-oncogenic role of Ecd in the progression of mammary oncogenesis through promoting cell survival.
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Directory of Open Access Journals (Sweden)
Hua Wang
Full Text Available Osteoporosis is a common disorder characterized by compromised bone strength that predisposes patients to increased fracture risk. Parathyroid hormone related protein (PTHrP is one of the candidates for clinical osteoporosis treatment. In this study, GST Gene Fusion System was used to express recombinant human PTHrP (hPTHrP 1-34 and 1-84. To determine whether the recombinant hPTHrP1-34 and 1-84 can enhance renal calcium reabsorption and promote bone formation, we examined effects of recombinant hPTHrP1-34 and 1-84 on osteogenic lineage commitment in a primary bone marrow cell culture system and on osteoporosis treatment. Results revealed that both of recombinant hPTHrP1-34 and 1-84 increased colony formation and osteogenic cell differentiation and mineralization in vitro; however, the effect of recombinant hPTHrP1-84 is a little stronger than that of hPTHrP1-34. Next, ovariectomy was used to construct osteoporosis animal model (OVX to test activities of these two recombinants in vivo. HPTHrP1-84 administration elevated serum calcium by up-regulating the expression of renal calcium transporters, which resulted in stimulation of osteoblastic bone formation. These factors contributed to augmented bone mass in hPTHrP1-84 treated OVX mice but did not affect bone resorption. There was no obvious bone mass alteration in hPTHrP1-34 treated OVX mice, which may be, at least partly, associated with shorter half-life of hPTHrP1-34 compared to hPTHrP1-84 in vivo. This study implies that recombinant hPTHrP1-84 is more effective than hPTHrP1-34 to enhance renal calcium reabsorption and to stimulate bone formation in vivo.
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Directory of Open Access Journals (Sweden)
Yinping Jiao
2018-01-01
Full Text Available Sorghum (Sorghum bicolor Moench, L. plant accumulates copious layers of epi-cuticular wax (EW on its aerial surfaces, to a greater extent than most other crops. EW provides a vapor barrier that reduces water loss, and is therefore considered to be a major determinant of sorghum's drought tolerance. However, little is known about the genes responsible for wax accumulation in sorghum. We isolated two allelic mutants, bloomless40-1 (bm40-1 and bm40-2, from a mutant library constructed from ethyl methane sulfonate (EMS treated seeds of an inbred, BTx623. Both mutants were nearly devoid of the EW layer. Each bm mutant was crossed to the un-mutated BTx623 to generated F2 populations that segregated for the bm phenotype. Genomic DNA from 20 bm F2 plants from each population was bulked for whole genome sequencing. A single gene, Sobic.001G228100, encoding a GDSL-like lipase/acylhydrolase, had unique homozygous mutations in each bulked F2 population. Mutant bm40-1 harbored a missense mutation in the gene, whereas bm40-2 had a splice donor site mutation. Our findings thus provide strong evidence that mutation in this GDSL-like lipase gene causes the bm phenotype, and further demonstrate that this approach of sequencing two independent allelic mutant populations is an efficient method for identifying causal mutations. Combined with allelic mutants, MutMap provides powerful method to identify all causal genes for the large collection of bm mutants in sorghum, which will provide insight into how sorghum plants accumulate such abundant EW on their aerial surface. This knowledge may facilitate the development of tools for engineering drought-tolerant crops with reduced water loss.
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Level structures in odd-odd deformed nucleus {sup 184}Ta
Energy Technology Data Exchange (ETDEWEB)
Gowrishankar, R.; Sood, P.C. [Sri Sathya Sai Institute of Higher Learning, Department of Physics, Prasanthinilayam (India)
2016-02-15
A detailed low-energy level scheme of the odd-odd n-rich nucleus {sub 73}{sup 184}Ta{sub 111} is constructed using the well tested Two-Quasiparticle Rotor Model (TQRM) for calculating the bandhead energies of physically admissible 2qp configurations with the inclusion of residual n-p interaction contribution. The presently available data on this level scheme from {sup 184}Hf decay are shown to be in agreement with these calculations. Our analysis concludes that {sup 184}Hf (Q{sub β} = 1340(30) keV) decay admits of 7 additional (to the 3 presently reported) β -branches to {sup 184}Ta with J = 0 or 1 and 8 more physically admissible weak (1f{sup u}) β -branches populating J{sup π} = 2{sup -} levels in {sup 184}Ta. Further, a close examination of our level scheme clearly indicates the existence of a low-lying (E{sub x} = 260(40) keV) high-spin (J{sup π} = 10{sup -}) long-lived isomer in this nucleus. (orig.)
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Genetics of leaf rust-resistant mutant WH 147-LM-1 in hexaploid wheat variety WH 147
International Nuclear Information System (INIS)
Reddy, V.R.K.; Viswanathan, P.
1999-01-01
By applying gamma rays, EMS and their combination in hexaploid wheat variety WH 147, a total of 20 mutants (0.0226%) exhibiting complete leaf rust resistance were isolated from segregating M2 rows.When one of the rust-resistant mutants, WH 147-LM-1 was crossed with the universally susceptible, suggesting that the mutant character is controlled by one dominant gene and one recessive gene.The F2 plants derived by crossing the mutant WH 147-LM with seven near-isogenic wheat lines showed segregation for susceptibility, indicating that the mutant character was indeed generated through induced mutations
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Directory of Open Access Journals (Sweden)
Khoa D Tran
Full Text Available In a variety of eukaryotes, flagella play important roles both in motility and as sensory organelles that monitor the extracellular environment. In the parasitic protozoan Leishmania mexicana, one glucose transporter isoform, LmxGT1, is targeted selectively to the flagellar membrane where it appears to play a role in glucose sensing. Trafficking of LmxGT1 to the flagellar membrane is dependent upon interaction with the KHARON1 protein that is located at the base of the flagellar axoneme. Remarkably, while Δkharon1 null mutants are viable as insect stage promastigotes, they are unable to survive as amastigotes inside host macrophages. Although Δkharon1 promastigotes enter macrophages and transform into amastigotes, these intracellular parasites are unable to execute cytokinesis and form multinucleate cells before dying. Notably, extracellular axenic amastigotes of Δkharon1 mutants replicate and divide normally, indicating a defect in the mutants that is only exhibited in the intra-macrophage environment. Although the flagella of Δkharon1 amastigotes adhere to the phagolysomal membrane of host macrophages, the morphology of the mutant flagella is often distorted. Additionally, these null mutants are completely avirulent following injection into BALB/c mice, underscoring the critical role of the KHARON1 protein for viability of intracellular amastigotes and disease in the animal model of leishmaniasis.
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Chen, Anli; Gao, Peng; Zhao, Qiaoling; Tang, Shunming; Shen, Xingjia; Zhang, Guozheng; Qiu, Zhiyong; Xia, Dingguo; Huang, Yongping; Xu, Yunmin; He, Ningjia
2013-02-25
The egg stage is an important stage in the silkworm (Bombyx mori) life cycle. Normal silkworm eggs are usually short, elliptical, and laterally flattened, with a sometimes hollowed surface on the lateral side. However, the eggs laid by homozygous recessive "Ming" lethal egg mutants (l-e(m)) lose water and become concaved around 1h, ultimately exhibiting a triangular shape on the egg surfaces. We performed positional cloning, and narrowed down the region containing the gene responsible for the l-e(m) mutant to 360 kb on chromosome 10 using 2287 F(2) individuals. Using expression analysis and RNA interference, the best l-e(m) candidate gene was shown to be BmEP80. The results of the inverse polymerase chain reaction showed that an ~1.9 kb region from the 3' untranslated region of BmVMP23 to the forepart of BmEP80 was replaced by a >100 kb DNA fragment in the l-e(m) mutant. Several eggs laid by the normal moths injected with BmEP80 small interfering RNAs were evidently depressed and exhibited a triangular shape on the surface. The phenotype exhibited was consistent with the eggs laid by the l-e(m) mutant. Moreover, two-dimensional gel electrophoresis showed that the BmEP80 protein was expressed in the ovary from the 9th day of the pupa stage to eclosion in the wild-type silkworm, but was absent in the l-e(m) mutant. These results indicate that BmEP80 is responsible for the l-e(m) mutation. Copyright © 2012 Elsevier B.V. All rights reserved.
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International Nuclear Information System (INIS)
Nagasaka, Takuya; Hishinuma, Yoshimitsu; Muroga, Takeo; Li, Yanfen; Watanabe, Hideo; Tanigawa, Hiroyasu; Sakasegawa, Hideo; Ando, Masami
2011-01-01
Extraction residue analysis was conducted on reduced activation ferritic/martensitic steels, such as F82H-BA07 heat, F82H-IEA heat, JLF-1 JOYO heat and CLAM steel. M 23 C 6 type precipitates, TaC precipitates and Fe 2 W Laves phase were identified in the present analyses. M 23 C 6 precipitates were coarsened in F82H-BA07 compared with the other steels at as-normalized and tempered (NT) condition. TaC precipitate formation was enhanced in JLF-1 and CLAM compared with F82H-BA07 and F82H-IEA at as-NT condition. Laves phase were detected in F82H-IEA after aging above 550 o C, where solid solution W was significantly decreased. F82H-IEA exhibited hardening after aging at 400 and 500 o C for 100 khr, whereas softening at 600 and 650 o C. This behavior is similar to JLF-1 and CLAM, and can be understood by precipitation of TaC and Laves phase.
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International Nuclear Information System (INIS)
Nath, S.; Gehlot, J.; Prasad, E.; Sadhukhan, Jhilam; Shidling, P.D.; Madhavan, N.; Muralithar, S.; Golda, K.S.; Jhingan, A.; Varughese, T.; Rao, P.V. Madhusudhana; Sinha, A.K.; Pal, Santanu
2011-01-01
We present γ-ray multiplicity distributions for the formation of evaporation residues in the fusion reaction 19 F + 184 W → 203 83 Bi 120 at beam energies in the range of 90-110 MeV. The measurements were carried out using a 14 element BGO detector array and the Heavy Ion Reaction Analyzer at the Inter University Accelerator Centre. The data have been unfolded to obtain angular momentum distributions with inputs from the statistical model calculation. Comparison with another neighboring system, viz. 19 F + 175 Lu → 194 80 Hg 114 with nearly similar entrance-channel mass asymmetry, hints at the depletion of higher angular momenta after crossing of the Z=82 shell in the compound nucleus.
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2010-04-01
... CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 184.1655 Propane. (a) Propane (empirical formula C3H8, CAS Reg. No. 74-98-6) is also known as dimethylmethane or propyl hydrid. It is a colorless, odorless, flammable gas at normal...
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Gao, Jian; Liang, Li; Chen, Qingqing; Zhang, Ling; Huang, Tonghui
2018-02-01
Acetyl-coenzyme A carboxylases (ACCs) is the first committed enzyme of fatty acid synthesis pathway. The inhibition of ACC is thought to be beneficial not only for diseases related to metabolism, such as type-2 diabetes, but also for infectious disease like bacterial infection disease. Soraphen A, a potent allosteric inhibitor of BC domain of yeast ACC, exhibit lower binding affinities to several yeast ACC mutants and the corresponding drug resistance mechanisms are still unknown. We report here a theoretical study of binding of soraphen A to wild type and yeast ACC mutants (including F510I, N485G, I69E, E477R, and K73R) via molecular dynamic simulation and molecular mechanics/generalized Born surface area free energy calculations methods. The calculated binding free energies of soraphen A to yeast ACC mutants are weaker than to wild type, which is highly consistent with the experimental results. The mutant F510I weakens the binding affinity of soraphen A to yeast ACC mainly by decreasing the van der Waals contributions, while the weaker binding affinities of Soraphen A to other yeast ACC mutants including N485G, I69E, E477R, and K73R are largely attributed to the decreased net electrostatic (ΔE ele + ΔG GB) interactions. Our simulation results could provide important insights for the development of more potent ACC inhibitors.
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Identification of Vitis vinifera L. grape berry skin color mutants and polyphenolic profile.
Ferreira, Vanessa; Fernandes, Fátima; Pinto-Carnide, Olinda; Valentão, Patrícia; Falco, Virgílio; Martín, Juan Pedro; Ortiz, Jesús María; Arroyo-García, Rosa; Andrade, Paula B; Castro, Isaura
2016-03-01
A germplasm set of twenty-five grapevine accessions, forming eleven groups of possible berry skin color mutants, were genotyped with twelve microsatellite loci, being eleven of them identified as true color mutants. The polyphenolic profiling of the confirmed mutant cultivars revealed a total of twenty-four polyphenols, comprising non-colored compounds (phenolic acids, flavan-3-ols, flavonols and a stilbene) and anthocyanins. Results showed differences in the contribution of malvidin-3-O-glucoside to the characteristic Pinot Noir anthocyanins profile. Regarding the two Pique-Poul colored variants, the lighter variant was richer than the darker one in all classes of compounds, excepting anthocyanins. In Moscatel Galego Roxo the F3'H pathway seems to be more active than F3'5'H, resulting in higher amounts of cyanidin, precursor of the cyanidin derivatives. As far as we are aware, this is the first time that a relationship between the content of polyphenolic compounds is established in groups of grape berry skin color mutant cultivars. Copyright © 2015 Elsevier Ltd. All rights reserved.
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Mutant IDH1 Promotes Glioma Formation In Vivo
Directory of Open Access Journals (Sweden)
Beatrice Philip
2018-05-01
Full Text Available Summary: Isocitrate dehydrogenase 1 (IDH1 is the most commonly mutated gene in grade II–III glioma and secondary glioblastoma (GBM. A causal role for IDH1R132H in gliomagenesis has been proposed, but functional validation in vivo has not been demonstrated. In this study, we assessed the role of IDH1R132H in glioma development in the context of clinically relevant cooperating genetic alterations in vitro and in vivo. Immortal astrocytes expressing IDH1R132H exhibited elevated (R-2-hydroxyglutarate levels, reduced NADPH, increased proliferation, and anchorage-independent growth. Although not sufficient on its own, IDH1R132H cooperated with PDGFA and loss of Cdkn2a, Atrx, and Pten to promote glioma development in vivo. These tumors resembled proneural human mutant IDH1 GBM genetically, histologically, and functionally. Our findings support the hypothesis that IDH1R132H promotes glioma development. This model enhances our understanding of the biology of IDH1R132H-driven gliomas and facilitates testing of therapeutic strategies designed to combat this deadly disease. : Philip et al. show that mutant IDH1 cooperates with PDGFA and loss of Cdkn2a, Atrx, and Pten to promote gliomagenesis in vivo in a mouse model of glioma. These tumors resemble proneural human mutant IDH1 glioblastoma and exhibit enhanced sensitivity to PARP inhibition in combination with chemotherapy. Keywords: IDH1, Cdkn2a, Atrx, Pten, glioma, mouse model, RCAS/TVA
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International Nuclear Information System (INIS)
Bresler, S.E.; Kalinin, V.L.; Laneeva, N.I.
1984-01-01
Radioresistant mutant Gam sup(r) 444 differs from a wild type and from Gam sup(r) 445 mutant in decreased frequency of long episome heritage ORF 1 (pur E + -tsx + -proC + -lac + ) and F 14 (ilv + -argE + ), containing hot points of RecRecF - depending recombination and in increased frequency of chromosome mobilization and integrative suppression of temperature sensitive dna A46 mutation by sexual factor F. In this respect Gam sup(r) 444 mutant resembles rec BC sbs B mutant with RecF - recombination type
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Directory of Open Access Journals (Sweden)
Qureshi Nasib
2006-05-01
Full Text Available Abstract Background The field of plasmid-based functional proteomics requires the rapid assay of proteins expressed from plasmid libraries. Automation is essential since large sets of mutant open reading frames are being cloned for evaluation. To date no integrated automated platform is available to carry out the entire process including production of plasmid libraries, expression of cloned genes, and functional testing of expressed proteins. Results We used a functional proteomic assay in a multiplexed setting on an integrated plasmid-based robotic workcell for high-throughput screening of mutants of cellulase F, an endoglucanase from the anaerobic fungus Orpinomyces PC-2. This allowed us to identify plasmids containing optimized clones expressing mutants with improved activity at lower pH. A plasmid library of mutagenized clones of the celF gene with targeted variations in the last four codons was constructed by site-directed PCR mutagenesis and transformed into Escherichia coli. A robotic picker integrated into the workcell was used to inoculate medium in a 96-well deep well plate, combining the transformants into a multiplexed set in each well, and the plate was incubated on the workcell. Plasmids were prepared from the multiplexed culture on the liquid handler component of the workcell and used for in vitro transcription/translation. The multiplexed expressed recombinant proteins were screened for improved activity and stability in an azo-carboxymethylcellulose plate assay. The multiplexed wells containing mutants with improved activity were identified and linked back to the corresponding multiplexed cultures stored in glycerol. Spread plates were prepared from the glycerol stocks and the workcell was used to pick single colonies from the spread plates, prepare plasmid, produce recombinant protein, and assay for activity. The screening assay and subsequent deconvolution of the multiplexed wells resulted in identification of improved CelF
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International Nuclear Information System (INIS)
Wu, Hsu-Pin; Hsu, Shu-Yuan; Wu, Wen-Ai; Hu, Ji-Wei; Ouyang, Pin
2014-01-01
Highlights: •Pnn CCD domain functions as a dominant negative mutant regulating Pnn expression and function. •Pnn CCD mutant Tg mice have a muscle wasting phenotype during development and show dystrophic histological features. •Pnn mutant muscles are susceptible to slow fiber type gene transition and NEB reduction. •The Tg mouse generated by overexpression of the Pnn CCD domain displays many characteristics resembling NEB +/− mice. -- Abstract: Pinin (Pnn) is a nuclear speckle-associated SR-like protein. The N-terminal region of the Pnn protein sequence is highly conserved from mammals to insects, but the C-terminal RS domain-containing region is absent in lower species. The N-terminal coiled-coil domain (CCD) is, therefore, of interest not only from a functional point of view, but also from an evolutionarily standpoint. To explore the biological role of the Pnn CCD in a physiological context, we generated transgenic mice overexpressing Pnn mutant in skeletal muscle. We found that overexpression of the CCD reduces endogenous Pnn expression in cultured cell lines as well as in transgenic skeletal muscle fibers. Pnn mutant mice exhibited reduced body mass and impaired muscle function during development. Mutant skeletal muscles show dystrophic histological features with muscle fibers heavily loaded with centrally located myonuclei. Expression profiling and pathway analysis identified over-representation of genes in gene categories associated with muscle contraction, specifically those related to slow type fiber. In addition nebulin (NEB) expression level is repressed in Pnn mutant skeletal muscle. We conclude that Pnn downregulation in skeletal muscle causes a muscular dystrophic phenotype associated with NEB deficiency and the CCD domain is incapable of replacing full length Pnn in terms of functional capacity
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Energy Technology Data Exchange (ETDEWEB)
Wu, Hsu-Pin; Hsu, Shu-Yuan [Department of Anatomy, Chang Gung University Medical College, Taiwan (China); Wu, Wen-Ai; Hu, Ji-Wei [Transgenic Mouse Core Laboratory, Chang Gung University, Taiwan (China); Ouyang, Pin, E-mail: ouyang@mail.cgu.edu.tw [Department of Anatomy, Chang Gung University Medical College, Taiwan (China); Transgenic Mouse Core Laboratory, Chang Gung University, Taiwan (China); Molecular Medicine Research Center, Chang Gung University, Taiwan (China)
2014-01-03
Highlights: •Pnn CCD domain functions as a dominant negative mutant regulating Pnn expression and function. •Pnn CCD mutant Tg mice have a muscle wasting phenotype during development and show dystrophic histological features. •Pnn mutant muscles are susceptible to slow fiber type gene transition and NEB reduction. •The Tg mouse generated by overexpression of the Pnn CCD domain displays many characteristics resembling NEB{sup +/−} mice. -- Abstract: Pinin (Pnn) is a nuclear speckle-associated SR-like protein. The N-terminal region of the Pnn protein sequence is highly conserved from mammals to insects, but the C-terminal RS domain-containing region is absent in lower species. The N-terminal coiled-coil domain (CCD) is, therefore, of interest not only from a functional point of view, but also from an evolutionarily standpoint. To explore the biological role of the Pnn CCD in a physiological context, we generated transgenic mice overexpressing Pnn mutant in skeletal muscle. We found that overexpression of the CCD reduces endogenous Pnn expression in cultured cell lines as well as in transgenic skeletal muscle fibers. Pnn mutant mice exhibited reduced body mass and impaired muscle function during development. Mutant skeletal muscles show dystrophic histological features with muscle fibers heavily loaded with centrally located myonuclei. Expression profiling and pathway analysis identified over-representation of genes in gene categories associated with muscle contraction, specifically those related to slow type fiber. In addition nebulin (NEB) expression level is repressed in Pnn mutant skeletal muscle. We conclude that Pnn downregulation in skeletal muscle causes a muscular dystrophic phenotype associated with NEB deficiency and the CCD domain is incapable of replacing full length Pnn in terms of functional capacity.
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Microstructural evolution of HFIR-irradiated low activation F82H and F82H-10B steels
International Nuclear Information System (INIS)
Wakai, E.; Shiba, K.; Sawai, T.; Hashimoto, N.; Robertson, J.P.; Klueh, R.L.
1998-01-01
Microstructures of reduced-activation F82H (8Cr-2W-0.2V-0.04Ta) and the F82H steels doped with 10 B, irradiated at 250 and 300 C to 3 and 57 dpa in the High Flux Isotope Reactor (HFIR), were examined by TEM. In the F82H irradiated at 250 C to 3 dpa, dislocation loops, small unidentified defect clusters with a high number density, and a few MC precipitates were observed in the matrix. The defect microstructure after 300 C irradiation to 57 dpa is dominated by the loops, and the number density of loops was lower than that of the F82H- 10 B steel. Cavities were observed in the F82H- 10 B steels, but the swelling value is insignificant. Small particles of M 6 C formed on the M 23 C 6 carbides that were present in both steels before the irradiation at 300 C to 57 dpa. A low number density of MC precipitate particles formed in the matrix during irradiation at 300 C to 57 dpa
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Pharmacokinetic properties of radiolabeled mutant Interleukin-2v: a PET imaging study.
Hartimath, Siddesh V; Manuelli, Valeria; Zijlma, Rolf; Signore, Alberto; Nayak, Tapan K; Freimoser-Grundschober, Anne; Klein, Christian; Dierckx, Rudi A J O; de Vries, Erik F J
2018-01-23
Interleukin-2 (IL2) is a cytokine that can stimulate cytotoxic immune cells to attack infected and malignant cells. Unfortunately, IL2 can also cause serious immune-related toxicity. Recently, a mutant of IL2 (IL2v) with abolished CD25 binding, increased plasma half-life and less toxicity was engineered. Unlike wild-type IL2 (wt-IL2), mutant IL2v does not bind to the α-subunit (CD25) of the high affinity IL2αβγ receptor, but only to its β and γ subunit. Here, we investigated the biological properties of IL2v and compared with the wt-IL2 using fluorine-18 and PET. [ 18 F]FB-IL2v binds specifically to IL2 receptors (IL2R) on activated human peripheral blood monocytes (hPBMCs) and is cleared mainly by the kidneys (Balb/c mice). [ 18 F]FB-IL2v PET studies in SCID mice injected with hPBMCs revealed high uptake in the implant (0.85 ± 0.15 SUV), which was significantly reduced after pretreatment with wt-IL2 or mutant IL2v (SUV 0.26 ± 0.1 and 0.46 ± 0.1, p FB-IL2v to IL2R was reversible. The volume of distribution (V T ) and the non-displaceable binding potential (BP nd ) of mutant [ 18 F]FB-IL2v in the implant were approximately 3 times lower than those of wild-type [ 18 F]FB-IL2 ( p FB-IL2v in the implant ( p FB-IL2 binds stronger to IL2R and has faster kinetics than [18F]FB-IL2v, which makes it less suitable as a therapeutic drug. [ 18 F]FB-IL2v, on the other hand, seems to have better properties for use as a therapeutic drug.
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Selection and agronomic evaluation of induced mutant lines of sesame
International Nuclear Information System (INIS)
Hoballah, A.A.
2001-01-01
%), capsules/plant (33.6%), capsule length (19.0%), 100-seed weight (18.6%) and plant height (12.1%). Analysis of variance for combining ability indicated that general (GCA) and specific (SCA) combining ability variances were highly significant for all studied traits. Estimates of GCA effects showed that EXM90, EFM92 and Mutant 8 were the best general combiners for earliness; the variety Giza 32 and Mutant 48 the best for seed yield and number of capsules per plant. Both parents and their derived crosses could be utilized for hybrid sesame production and varietal improvement in terms of the probability of selecting desirable segregants for yield and yield components. Estimates of the type of gene action confirmed the importance of both additive and non-additive (dominant) gene effects in the inheritance of the studied characters in both F 1 and F 2 generations. However the dominance components were larger than the additive ones for most investigated traits in the F 1 and vice versa in the F 2 . Overdominance was also noted. Heritability in narrow sense (H n ) was low for most characters in the F 1 . On the other hand, high values of heritability (H n ) were recorded for all the investigated traits in the F 2 , indicating that the genetic variance associated with those traits was mostly due to additive effects of genes, and therefore, it could be concluded that selection based on the accumulation of additive effects would be very successful in improving such traits. (author)
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Hemin reconstitutes proton extrusion in an H+-ATPase-negative mutant of Lactococcus lactis
DEFF Research Database (Denmark)
Blank, L.M.; Købmann, Brian Jensen; Michelsen, Ole
2001-01-01
H+-ATPase is considered essential for growth of Lactococcus lactis. However, media containing hemin restored the aerobic growth of an H+-ATPase-negative mutant, suggesting that hemin complements proton extrusion. We show that inverted membrane vesicles prepared from hemin-grown L. lactis cells...
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Federal Laboratory Consortium — The F/H Laboratory is the largest of the eight laboratories within the SRS Analytical Area Project with over 100,000 square feet of floor space, 20,000 square foot...
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International Nuclear Information System (INIS)
Kuila, R.K.; Ranganathan, B.
1978-01-01
A strain of Streptococcus lactis subspecies diacetylactis S 1 isolated from fresh milk was exposed to 7200 ergs/mm 2 of ultraviolet radiation. Over 8100 colonies surviving from 7.4 x 10 6 cells exposed to radiation were screened on citrate agar for detection and isolation of mutants with increased flavor and/or acid production. Of the survivors, 960 were type-I mutants that exhibited clear zone on citrate agar after 18 h (presumed to be high diacetyl producers), and 288 were type-II mutants which did not exhibit clear zones on citrate agar for up to 72 h (high acid producers). Type-II mutants produced an average .93 percent titratable acidity which was 34 percent more than the .69 percent of the parent. Reduction in titratable acidity (56 percent less) was considerable in type-I mutants, compared with the parent culture. Diacetyl + acetoin production by type-I mutants was 137.9 ppM which has 4.5 times more than that of the parental strain. Acetaldehyde production in the mutants varied from 1.5 to 34.5 ppM (parent culture 3.0 ppM). The mutants with increased acid and high acetoin plus diacetyl production were stable after 50 subcultures in milk
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Isolation and characterization of mutant strains of Escherichia coli altered in H2 metabolism
International Nuclear Information System (INIS)
Lee, J.H.; Patel, P.; Sankar, P.; Shanmugam, K.T.
1985-01-01
A positive selection procedure is described for the isolation of hydrogenase-defective mutant strains of Escherichia coli. Mutant strains isolated by this procedure can be divided into two major classes. Class II mutants produced hydrogenase activity (determined by using a tritium-exchange assay) and formate hydrogenlyase activity but lacked the ability to reduce benzyl viologen or fumarate with H 2 as the electron donor. Class I mutants failed to produce active hydrogenase and hydrogenase-dependent activities. All the mutant strains produced detectable levels of formate dehydrogenase-1 and -2 and fumarate reductase. The mutation in class I mutants mapped near 65 min of the E. coli chromosome, whereas the mutation in class II mutants mapped between srl and cys operons (58 and 59 min, respectively) in the genome. The class II Hyd mutants can be further subdivided into two groups (hydA and hydB) based on the cotransduction characteristics with cys and srl. These results indicate that there are two hyd operons and one hup operon in the E. coli chromosome. The two hyd operons are needed for the production of active hydrogenase, and all three are essential for hydrogen-dependent growth of the cell
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Directory of Open Access Journals (Sweden)
David Wifling
Full Text Available In contrast to the corresponding mouse and rat orthologs, the human histamine H4 receptor (hH4R shows extraordinarily high constitutive activity. In the extracellular loop (ECL, replacement of F169 by V as in the mouse H4R significantly reduced constitutive activity. Stabilization of the inactive state was even more pronounced for a double mutant, in which, in addition to F169V, S179 in the ligand binding site was replaced by M. To study the role of the FF motif in ECL2, we generated the hH4R-F168A mutant. The receptor was co-expressed in Sf9 insect cells with the G-protein subunits Gαi2 and Gβ1γ2, and the membranes were studied in [3H]histamine binding and functional [35S]GTPγS assays. The potency of various ligands at the hH4R-F168A mutant decreased compared to the wild-type hH4R, for example by 30- and more than 100-fold in case of the H4R agonist UR-PI376 and histamine, respectively. The high constitutive activity of the hH4R was completely lost in the hH4R-F168A mutant, as reflected by neutral antagonism of thioperamide, a full inverse agonist at the wild-type hH4R. By analogy, JNJ7777120 was a partial inverse agonist at the hH4R, but a partial agonist at the hH4R-F168A mutant, again demonstrating the decrease in constitutive activity due to F168A mutation. Thus, F168 was proven to play a key role not only in ligand binding and potency, but also in the high constitutive activity of the hH4R.
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Ji, S H; Gururani, M A; Lee, J W; Ahn, B-O; Chun, S-C
2014-03-01
We have isolated a severe dwarf mutant derived from a Ds (Dissociation) insertion mutant rice (Oryza sativa var. japonica c.v. Dongjin). This severe dwarf phenotype, has short and dark green leaves, reduced shoot growth early in the seedling stage, and later severe dwarfism with failure to initiate flowering. When treated with bioactive GA3 , mutants are restored to the normal wild-type phenotype. Reverse transcription PCR analyses of 22 candidate genes related to the gibberellin (GA) biosynthesis pathway revealed that among 22 candidate genes tested, a dwarf mutant transcript was not expressed only in one OsKS2 gene. Genetic analysis revealed that the severe dwarf phenotype was controlled by recessive mutation of a single nuclear gene. The putative OsKS2 gene was a chromosome 4-located ent-kaurene synthase (KS), encoding the enzyme that catalyses an early step of the GA biosynthesis pathway. Sequence analysis revealed that osks2 carried a 1-bp deletion in the ORF region of OsKS2, which led to a loss-of-function mutation. The expression pattern of OsKS2 in wild-type cv Dongjin, showed that it is expressed in all organs, most prominently in the stem and floral organs. Morphological characteristics of the dwarf mutant showed dramatic modifications in internal structure and external morphology. We propose that dwarfism in this mutant is caused by a point mutation in OsKS2, which plays a significant role in growth and development of higher plants. Further investigation on OsKS2 and other OsKS-like proteins is underway and may yield better understanding of the putative role of OsKS in severe dwarf mutants. © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands.
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Directory of Open Access Journals (Sweden)
Maria José Menal
2018-01-01
Full Text Available BackgroundEvidence from patients and animal models suggests that obstructive sleep apnea (OSA may increase the risk of Alzheimer’s disease (AD and that AD is associated with reduced brain tissue stiffness.AimTo investigate whether intermittent hypoxia (IH alters brain cortex tissue stiffness in AD mutant mice exposed to IH mimicking OSA.MethodsSix-eight month old (B6C3-Tg(APPswe,PSEN1dE985Dbo/J AD mutant mice and wild-type (WT littermates were subjected to IH (21% O2 40 s to 5% O2 20 s; 6 h/day or normoxia for 8 weeks. After euthanasia, the stiffness (E of 200-μm brain cortex slices was measured by atomic force microscopy.ResultsTwo-way ANOVA indicated significant cortical softening and weight increase in AD mice compared to WT littermates, but no significant effects of IH on cortical stiffness and weight were detected. In addition, reduced myelin was apparent in AD (vs. WT, but no significant differences emerged in the cortex extracellular matrix components laminin and glycosaminoglycans when comparing baseline AD and WT mice.ConclusionAD mutant mice exhibit reduced brain tissue stiffness following both normoxia and IH mimicking sleep apnea, and such differences are commensurate with increased edema and demyelination in AD.
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Study of the decay 184Hg→184Au (Tsub(1/2)=30s)
International Nuclear Information System (INIS)
Nettles, W.G.; Beraud, R.; Cole, J.D.; Hamilton, J.H.; Ramayya, A.V.; Kawakami, H.; Spejewski, E.H.
1978-01-01
The radioactive decay of 184 Hg to 184 Au was studied on line with the UNISOR isotope separator. The 184 mass chain was entered at 184 Tl and 184 Hg by bombarding an isotopically-enriched target of 180 W with 14 N ions of 177 MeV. Multiscale singles and γ-γ-time coincidence studies were carried out. No previous levels have been assigned to 184 Au. From the coincidence data eight levels at 156.2, 159.1, 247.7, 282.8, 295.1, 392.4, 418.0 and 412.4 keV are assigned and two tentative levels at 534.0 and 551.6 keV are proposed. By using the delayed coincidence technique, a half-life of 36+-6 nanoseconds was measured for the 156.2 keV first-excited level and upper limits of 2 nanoseconds were determined for the half-lives of levels at 159.1, 247.7, 295.1 and 392.4 keV [fr
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Rubisco mutants of Chlamydomonas reinhardtii enhance photosynthetic hydrogen production.
Pinto, T S; Malcata, F X; Arrabaça, J D; Silva, J M; Spreitzer, R J; Esquível, M G
2013-06-01
Molecular hydrogen (H2) is an ideal fuel characterized by high enthalpy change and lack of greenhouse effects. This biofuel can be released by microalgae via reduction of protons to molecular hydrogen catalyzed by hydrogenases. The main competitor for the reducing power required by the hydrogenases is the Calvin cycle, and rubisco plays a key role therein. Engineered Chlamydomonas with reduced rubisco levels, activity and stability was used as the basis of this research effort aimed at increasing hydrogen production. Biochemical monitoring in such metabolically engineered mutant cells proceeded in Tris/acetate/phosphate culture medium with S-depletion or repletion, both under hypoxia. Photosynthetic activity, maximum photochemical efficiency, chlorophyll and protein levels were all measured. In addition, expression of rubisco, hydrogenase, D1 and Lhcb were investigated, and H2 was quantified. At the beginning of the experiments, rubisco increased followed by intense degradation. Lhcb proteins exhibited monomeric isoforms during the first 24 to 48 h, and D1 displayed sensitivity under S-depletion. Rubisco mutants exhibited a significant decrease in O2 evolution compared with the control. Although the S-depleted medium was much more suitable than its complete counterpart for H2 production, hydrogen release was observed also in sealed S-repleted cultures of rubisco mutated cells under low-moderate light conditions. In particular, the rubisco mutant Y67A accounted for 10-15-fold higher hydrogen production than the wild type under the same conditions and also displayed divergent metabolic parameters. These results indicate that rubisco is a promising target for improving hydrogen production rates in engineered microalgae.
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The application of shortened upper leaf mutant in barley breeding
International Nuclear Information System (INIS)
Jin Hua
2004-01-01
The shortened upper leaf mutant was induced from Fuji Nigo by γ-ray irradiation. Fuji Nigo, the mutant, cross-cut F 1 , F 2 and back-cross F 1 , F 2 were used to analyze mutant heredity by comparative study. The yield, chlorophyll content, light intensity, dry matter of mutant were investigated. The results showed that (1) the mutant character was controlled by a couple of nuclear genes which were partial dominance; (2) the transmittance of the mutant colony was better than that of Fuji Nigo and bottom dry matter was much more than that of Fuji Nigo; (3) under the condition of high fertilizer and high plant population , the yield of mutant was higher than that of Fuji Nigo; (4) the content of chlorophyll a in the mutant was higher than that in Fuji Nigo
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A molecular chaperone activity of CCS restores the maturation of SOD1 fALS mutants.
Luchinat, Enrico; Barbieri, Letizia; Banci, Lucia
2017-12-12
Superoxide dismutase 1 (SOD1) is an important metalloprotein for cellular oxidative stress defence, that is mutated in familiar variants of Amyotrophic Lateral Sclerosis (fALS). Some mutations destabilize the apo protein, leading to the formation of misfolded, toxic species. The Copper Chaperone for SOD1 (CCS) transiently interacts with SOD1 and promotes its correct maturation by transferring copper and catalyzing disulfide bond formation. By in vitro and in-cell NMR, we investigated the role of the SOD-like domain of CCS (CCS-D2). We showed that CCS-D2 forms a stable complex with zinc-bound SOD1 in human cells, that has a twofold stabilizing effect: it both prevents the accumulation of unstructured mutant SOD1 and promotes zinc binding. We further showed that CCS-D2 interacts with apo-SOD1 in vitro, suggesting that in cells CCS stabilizes mutant apo-SOD1 prior to zinc binding. Such molecular chaperone function of CCS-D2 is novel and its implications in SOD-linked fALS deserve further investigation.
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Energy Technology Data Exchange (ETDEWEB)
Lu, Yuan; Zhao, Hongxin; Zhang, Chong; Lai, Qiheng; Xing, Xin-Hui [Institute of Biochemical Engineering, Department of Chemical Engineering, Tsinghua University, Beijing 100084 (China)
2009-06-15
To examine perturbation effects of formate pathway on hydrogen productivity in Enterobacter aerogenes (Ea), formate dehydrogenase FDH-H gene (fdhF) and formate hydrogen lyase activator protein FHLA gene (fhlA) originated from Escherichia coli, were overexpressed in the wild strain Ea, its hycA-deleted mutant (A) by knockout the formate hydrogen lyase repressor and hybO-deleted mutant (O) by knockout of the uptake hydrogenase, respectively. Overexpression of fdhF and fhlA promoted cell growth and volumetric hydrogen production rates of all the strains, and the hydrogen production per gram cell dry weight (CDW) for Ea, A and O was increased by 38.5%, 21.8% and 5.25%, respectively. The fdhF and fhlA overexpression improved the hydrogen yield per mol glucose of strains Ea and A, but declined that of strain O. The increase of hydrogen yield of the strain Ea with fdhF and fhlA expression was mainly attributed to the increase of formate pathway, while for the mutant A, the improved hydrogen yield with fdhF and fhlA expression was mainly due to the increase of NADH pathway. Analysis of the metabolites and ratio of ethanol-to-acetate showed that the cellular redox state balance and energy level were also changed for these strains by fdhF and fhlA expression. These findings demonstrated that the hydrogen production was not only dependent on the hydrogenase genes, but was also affected by the regulation of the whole metabolism. Therefore, fdhF and fhlA expression in different strains of E. aerogenes could exhibit different perturbation effects on the metabolism and the hydrogen productivity. (author)
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Eom, Jeong Seon; Lee, Sun Young; Choi, Hye Sun
2014-11-01
Bacillus subtilis HJ18-4 isolated from buckwheat sokseongjang, a traditional Korean fermented soybean food, exhibits broad-spectrum antimicrobial activity against foodborne pathogens, including Bacillus cereus. In this study, we investigated the antibacterial efficacy and regulation of toxin gene expression in B. cereus by B. subtilis HJ18-4. Expression of B. cereus toxin-related genes (groEL, nheA, nheC, and entFM) was downregulated by B. subtilis HJ18-4, which also exhibited strong antibacterial activity against B. cereus. We also found that water extracts of soy product fermented with B. subtilis HJ18-4 significantly inhibited the growth of B. cereus and toxin expression. These results indicate that B. subtilis HJ18-4 could be used as an antimicrobial agent to control B. cereus in the fermented soybean food industry. Our findings also provide an opportunity to develop an efficient biological control agent against B. cereus. © 2014 The Authors. Journal of Food Science published by Wiley Periodicals, Inc. on behalf of Institute of Food Technologists®
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Christensen, L L; Jensen, U B; Bross, P; Orntoft, T F
2000-09-01
The alpha1,3/4-fucosyltransferases are involved in the synthesis of fucosylated cell surface glycoconjugates. Human alpha1,3/4-fucosyltransferase III, -V, and -VI (hFucTIII, -V, and -VI) contain two conserved C-terminal N-glycosylation sites (hFucTIII: Asn154 and Asn185; hFucTV: Asn167 and Asn198; and hFucTVI: Asn153 and Asn184). In the present study, we have analyzed the functional role of these potential N-glycosylation sites, laying the main emphasis on the sites in hFucTIII. Tunicamycin treatment completely abolished hFucTIII enzyme activity while castanospermine treatment diminished hFucTIII enzyme activity to approximately 40% of the activity of the native enzyme. To further analyze the role of the conserved N-glycosylation sites in hFucTIII, -V, and -VI, we made a series of mutant genomic DNAs in which the asparagine residues in the potential C-terminal N-glycosylation sites were replaced by glutamine. Subsequently, the hFucTIII, -V, and -VI wild type and the mutants were expressed in COS-7 cells. All the mutants exhibited lower enzyme activity than the wild type and elimination of individual sites had different effects on the activity. The mutations did not affect the protein level of the mutants in the cells, but reduced the molecular mass as predicted. Kinetic analysis of hFucTIII revealed that lack of glycosylation at Asn185 did not change the Km values for the oligosaccharide acceptor and the nucleotide sugar donor. The present study demonstrates that hFucTIII, -V, and -VI require N-glycosylation at the two conserved C-terminal N-glycosylation sites for expression of full enzyme activity.
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Zhou, Mingxu; Duan, Qiangde; Zhu, Xiaofang; Guo, Zhiyan; Li, Yinchau; Hardwidge, Philip R; Zhu, Guoqiang
2013-05-13
The role of flagella in the pathogenesis of F4ac+ Enterotoxigenic Escherichia coli (ETEC) mediated neonatal and post-weaning diarrhea (PWD) is not currently understood. We targeted the reference C83902 ETEC strain (O8:H19:F4ac+ LT+ STa+ STb+), to construct isogenic mutants in the fliC (encoding the major flagellin protein), motA (encoding the flagella motor), and faeG (encoding the major subunit of F4 fimbriae) genes. Both the ΔfliC and ΔfaeG mutants had a reduced ability to adhere to porcine intestinal epithelial IPEC-J2 cells. F4 fimbriae expression was significantly down-regulated after deleting fliC, which revealed that co-regulation exists between flagella and F4 fimbriae. However, there was no difference in adhesion between the ΔmotA mutant and its parent strain. These data demonstrate that both flagella and F4 fimbriae are required for efficient F4ac+ ETEC adhesion in vitro.
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Morphology, physiology, genetics, enigmas, and status of an extremely rare tree: Mutant tanoak
Philip M. McDonald; Jianwei Zhang; Randy S. Senock; Jessica W. Wright
2013-01-01
Important physical characteristics, morphological attributes, physiological functions, and genetic properties of mutant tanoak, Notholithocarpus densiflorus f. attenuato-dentatus (Fagaceae), and normal tanoak, Notholithocarpus densiflorus (Hook. & Arn.) Manos, Cannon & S. H. Oh, were studied on the Challenge...
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Structural Basis for a Switch in Receptor Binding Specificity of Two H5N1 Hemagglutinin Mutants
Directory of Open Access Journals (Sweden)
Xueyong Zhu
2015-11-01
Full Text Available Avian H5N1 influenza viruses continue to spread in wild birds and domestic poultry with sporadic infection in humans. Receptor binding specificity changes are a prerequisite for H5N1 viruses and other zoonotic viruses to be transmitted among humans. Previous reported hemagglutinin (HA mutants from ferret-transmissible H5N1 viruses of A/Vietnam/1203/2004 and A/Indonesia/5/2005 showed slightly increased, but still very weak, binding to human receptors. From mutagenesis and glycan array studies, we previously identified two H5N1 HA mutants that could more effectively switch receptor specificity to human-like α2-6-linked sialosides with avidity comparable to wild-type H5 HA binding to avian-like α2-3-linked sialosides. Here, crystal structures of these two H5 HA mutants free and in complex with human and avian glycan receptor analogs reveal the structural basis for their preferential binding to human receptors. These findings suggest continuous surveillance should be maintained to monitor and assess human-to-human transmission potential of H5N1 viruses.
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International Nuclear Information System (INIS)
Thoms, B.; Wackernagel, W.
1988-01-01
Mutations in recA, such as recA801(Srf) (suppressor of RecF) or recA441(Tif) (temperature-induced filamentation) partially suppress the deficiency in postreplication repair of UV damage conferred by recF mutations. We observed that spontaneous recA(Srf) mutants accumulated in cultures of recB recC sbcB sulA::Mu dX(Ap lac) lexA51 recF cells because they grew faster than the parental strain. We show that in a uvrA recB+ recC+ genetic background there are two prerequisites for the suppression by recA(Srf) of the UV-sensitive phenotype of recF mutants. (i) The recA(Srf) protein must be provided in increased amounts either by SOS derepression or by a recA operator-constitutive mutation in a lexA(Ind) (no induction of SOS functions) genetic background. (ii) The gene recJ, which has been shown previously to be involved in the recF pathway of recombination and repair, must be functional. The level of expression of recJ in a lexA(Ind) strain suffices for full suppression. Suppression by recA441 at 30 degrees C also depends on recJ+. The hampered induction by UV of the SOS gene uvrA seen in a recF mutant was improved by a recA(Srf) mutation. This improvement did not require recJ+. We suggest that recA(Srf) and recA(Tif) mutant proteins can operate in postreplication repair independent of recF by using the recJ+ function
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Ibrutinib targets mutant-EGFR kinase with a distinct binding conformation.
Wang, Aoli; Yan, Xiao-E; Wu, Hong; Wang, Wenchao; Hu, Chen; Chen, Cheng; Zhao, Zheng; Zhao, Peng; Li, Xixiang; Wang, Li; Wang, Beilei; Ye, Zi; Wang, Jinhua; Wang, Chu; Zhang, Wei; Gray, Nathanael S; Weisberg, Ellen L; Chen, Liang; Liu, Jing; Yun, Cai-Hong; Liu, Qingsong
2016-10-25
Ibrutinib, a clinically approved irreversible BTK kinase inhibitor for Mantle Cell Lymphoma (MCL) and Chronic Lymphocytic Leukemia (CLL) etc, has been reported to be potent against EGFR mutant kinase and currently being evaluated in clinic for Non Small Cell Lung Cancer (NSCLC). Through EGFR wt/mutant engineered isogenic BaF3 cell lines we confirmed the irreversible binding mode of Ibrutinib with EGFR wt/mutant kinase via Cys797. However, comparing to typical irreversible EGFR inhibitor, such as WZ4002, the washing-out experiments revealed a much less efficient covalent binding for Ibrutinib. The biochemical binding affinity examination in the EGFR L858R/T790M kinase revealed that, comparing to more efficient irreversible inhibitor WZ4002 (Kd: 0.074 μM), Ibrutinib exhibited less efficient binding (Kd: 0.18 μM). An X-ray crystal structure of EGFR (T790M) in complex with Ibrutinib exhibited a unique DFG-in/c-Helix-out inactive binding conformation, which partially explained the less efficiency of covalent binding and provided insight for further development of highly efficient irreversible binding inhibitor for the EGFR mutant kinase. These results also imply that, unlike the canonical irreversible inhibitor, sustained effective concentration might be required for Ibrutinib in order to achieve the maximal efficacy in the clinic application against EGFR driven NSCLC.
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International Nuclear Information System (INIS)
Morton, Craig J.; Cameron, Rachel; Lawrence, Lynne J.; Lin Bo; Lowe, Melinda; Luttick, Angela; Mason, Anthony; McKimm-Breschkin, Jenny; Parker, Michael W.; Ryan, Jane; Smout, Michael; Sullivan, Jayne; Tucker, Simon P.; Young, Paul R.
2003-01-01
Respiratory syncytial virus (RSV) is a ubiquitous human pathogen and the leading cause of lower respiratory tract infections in infants. Infection of cells and subsequent formation of syncytia occur through membrane fusion mediated by the RSV fusion protein (RSV-F). A novel in vitro assay of recombinant RSV-F function has been devised and used to characterize a number of escape mutants for three known inhibitors of RSV-F that have been isolated. Homology modeling of the RSV-F structure has been carried out on the basis of a chimera derived from the crystal structures of the RSV-F core and a fragment from the orthologous fusion protein from Newcastle disease virus (NDV). The structure correlates well with the appearance of RSV-F in electron micrographs, and the residues identified as contributing to specific binding sites for several monoclonal antibodies are arranged in appropriate solvent-accessible clusters. The positions of the characterized resistance mutants in the model structure identify two promising regions for the design of fusion inhibitors
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Käfer, E; Mayor, O
1986-07-01
To identify genes which affect DNA repair and possibly recombination in Aspergillus nidulans, mutants hypersensitive to methyl methanesulphonate (MMS) were induced with ultraviolet light (UV) or gamma-rays. About half of them contained associated translocations and many were hypersensitive to UV and/or defective in meiosis. Two are alleles of the known uvsB gene while most others define new genes. In addition, among available uvs mutants many were found to be MMS-sensitive. Some of the various uncharacterized ones were identified as alleles of known uvs, but 5 of them were mapped in 2 new genes, uvsH and uvsJ. To identify functional and epistatic groups, mutants from each uvs gene were tested for effects on recombination and mutation, and double mutant uvs strains were compared for UV survival to their component single mutant strains. 3 epistatic pairs were identified, (1) uvsF and H, (2) uvsB and D, and (3) uvsC and E. Conclusive interpair tests were difficult, because such double mutant combinations were frequently lethal or nearly so. The first pair, uvsF and H, shared some of the properties of excision-defective mutants, both uvs being very highly sensitive to UV for mutation as well as survival. But unlike such mutants, uvsH was also sensitive to gamma-rays and defective in meiosis. Both uvs showed normal levels of meiotic recombination, but greatly increased spontaneous mitotic crossing-over, being the most "hyperrec" types among all uvs. The second pair, uvsB and uvsC, which was similarly hyperrec showed only slight increases of UV-induced mutation (less than 2-fold). As a main effect, these uvs caused very high frequencies of unbalanced, unstable segregants from diploid conidia (30 X), but few of these were recognizable aneuploids. The third pair, uvsC and E, which are known to be rec- for gene conversion, caused reduced mitotic crossing-over in diploids and increased levels of haploid segregants. These mutants are spontaneous mutators, but showed less UV
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Preliminary research on morphological differentiation of avilamycin high-yield mutant strain H15
International Nuclear Information System (INIS)
Liang Xinle; Jin Yingyan; Chen Ming; Zhang Hong
2010-01-01
Morphological differentiation characters such as colony, sporotrichial, and conidiophores of mutant H15, which was derived from Streptomyces viridochromogenes 4.1119 treated with 60 Co γ-rays irradiation, were investigated by scanning electron microscope and fluorescence microscope. The results showed that mutant H15 was remarkable variation from the strain 4.1119. Cultured on agar surface, H15 had a grayish-whitish-green colony, linear sporotrichial, smooth and round conidiophore without any spike, whereas strain 4.1119 had spiral sporotrichial and round conidiophore with spike on the surface. In the submerged cultures, differentiation process of mycelia pellet of H15 was also different. Spores germinated as a compartmentalized mycelium, the young compartmentalized mycelium started to form pellets which grew in a radial pattern. After apoptosis took place in the center of the pellets, the pellet diameter growth arrested. Compared with the strain 4.1119, H15 required a long developing course for hyphae clustering and pellets formation (at 48 h, φ 245 μm). The stage of pellet arrest or apoptosis in the pellet centre were extended, which would benefit the avilamycin accumulation since the antibiotic was mainly produced at the same time. These suggested that pellet formation kinetics, relational balance between pellet diameter enlargement and mycelia apoptosis in the pellet arrest stage were key factors to avilamyin accumulation in submerged cultures of Streptomyces viridoehrongenes H15. (authors)
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Samuele, Alberta; Facchini, Marcella; Rotili, Dante; Mai, Antonello; Artico, Marino; Armand-Ugón, Mercedes; Esté, José A; Maga, Giovanni
2008-09-01
We recently reported the synthesis and biological evaluation of a novel series of 5-alkyl-2-(N,N-disubstituted)amino-6-(2,6-difluorophenylalkyl)-3,4-dihydropyrimidin-4(3H)-ones (F(2)-N,N-DABOs). These compounds are highly active against both wild-type HIV-1 and the K103N, Y181C, and Y188L mutant strains. Herein we present novel 6-(2-chloro-6-fluorophenylalkyl)-N,N-DABO (2-Cl-6-F-N,N-DABO) derivatives and investigate the molecular basis for their high-affinity binding to HIV-1 reverse transcriptase (RT). Our results show that the new compounds display higher association rates than the difluoro derivatives toward wild-type HIV-1 RT or drug-resistant RT mutant forms. We also show that they preferentially associate to either the free enzyme or the enzyme-nucleic acid binary complex, and that this binding is stabilized upon formation of the ternary complex between HIV-1 RT and both the nucleic acid and nucleotide substrates. Interestingly, one compound showed dissociation rates from the ternary complex with RT mutants K103N and Y181I 10-20-fold slower than from the corresponding complex with wild-type RT.
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Investigating the structure and dynamics of the PIK3CA wild-type and H1047R oncogenic mutant.
Directory of Open Access Journals (Sweden)
Paraskevi Gkeka
2014-10-01
Full Text Available The PIK3CA gene is one of the most frequently mutated oncogenes in human cancers. It encodes p110α, the catalytic subunit of phosphatidylinositol 3-kinase alpha (PI3Kα, which activates signaling cascades leading to cell proliferation, survival, and cell growth. The most frequent mutation in PIK3CA is H1047R, which results in enzymatic overactivation. Understanding how the H1047R mutation causes the enhanced activity of the protein in atomic detail is central to developing mutant-specific therapeutics for cancer. To this end, Surface Plasmon Resonance (SPR experiments and Molecular Dynamics (MD simulations were carried out for both wild-type (WT and H1047R mutant proteins. An expanded positive charge distribution on the membrane binding regions of the mutant with respect to the WT protein is observed through MD simulations, which justifies the increased ability of the mutated protein variant to bind to membranes rich in anionic lipids in our SPR experiments. Our results further support an auto-inhibitory role of the C-terminal tail in the WT protein, which is abolished in the mutant protein due to loss of crucial intermolecular interactions. Moreover, Functional Mode Analysis reveals that the H1047R mutation alters the twisting motion of the N-lobe of the kinase domain with respect to the C-lobe and shifts the position of the conserved P-loop residues in the vicinity of the active site. These findings demonstrate the dynamical and structural differences of the two proteins in atomic detail and propose a mechanism of overactivation for the mutant protein. The results may be further utilized for the design of mutant-specific PI3Kα inhibitors that exploit the altered mutant conformation.
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Corrosion fatigue studies on F82H mod. martensitic steel in reducing water coolant environments
Energy Technology Data Exchange (ETDEWEB)
Maday, M F; Masci, A [ENEA, Casaccia (Italy). Centro Ricerche Energia
1998-03-01
Load-controlled low cycle fatigue tests have been carried out on F82H martensitic steel in 240degC oxygen-free water with and without dissolved hydrogen, in order to simulate realistic coolant boundary conditions to be approached in DEMO. It was found that water independently of its hydrogen content, determined the same fatigue life reduction compared to the base-line air results. Water cracks exhibited in their first propagation stages similar fracture morphologies which were completely missing on the air cracks, and were attributed to the action of an environment related component. Lowering frequency gave rise to an increase in F82H fatigue lifetimes without any change in cracking mode in air, and to fatigue life reduction by microvoid coalescence alone in water. The data were discussed in terms of (i) frequency dependent concurrent processes for crack initiation and (ii) frequency-dependent competitive mechanisms for crack propagation induced by cathodic hydrogen from F82H corrosion. (author)
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The effect of UVB on flavonoid biosynthesis in wild type and mutant petunia and arabidopsis
International Nuclear Information System (INIS)
Ryan, K.G.; Swinny, E.E.; Markham, K.R.; Winefield, C.
2000-01-01
Full text: Flavonoids may protect plants against damage by UVB radiation. Flavonoid composition and mRNA expression were determined following growth of plants under natural light, and under natural light with low UVB and with enhanced UVB. In wild-type Arabidopsis and Petunia, UVB induced an increase in total levels of flavonols and this was due to an up-regulation of, several genes coding for key enzymes in the phenylpropanoid pathway. In addition, UVB induced a higher rate of production of the di-hydroxylated si flavonol, quercetin glycoside than of the mono-hydroxylated equivalent, of kaempferol glycoside. Thus the ratio of quercetin to kaempferol increased with UVB treatment in wild type plants, and this suggests that the flavonoid r 3'hydroxylase (F3'H) enzyme, which converts dihydrokaempferol to dihydroquercetin, may play a key role in plant protection from UVB. Mutant plants of both species lacking this F3'H gene were grown under similar UV conditions. Leaves of the mutant Arabidopsis plant (tt7) did not contain quercetin, even under the enhanced UVB treatment. Under the low UVB treatment the total amount of flavonol was similar to the wild-type (Ler), but with increasing UVB, total flavonol (i.e. kaempferol) levels were significantly higher than in similarly treated wild type plants. In the Petunia F3'H mutant, low levels of quercetin were found even in the low UVB treatment, which indicates this variety may be producing some quercetin via an alternative pathway. Under UVB radiation, total flavonoids increased to levels significantly higher than in similarly treated wild type plants, and most of this material was kaempferol. These observations suggest that quercetin is the preferred protective flavonol in wild type plants, due perhaps to enhanced antioxidant or free radical scavenging activity. In mutant plants lacking the F3'H enzyme, the response is to produce a larger amount of a less effective photoprotectant
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Mutations in RCA1 and AFG3 inhibit F1-ATPase assembly in Saccharomyces cerevisiae.
Paul, M F; Tzagoloff, A
1995-10-02
The RCA1 (YTA12) and AFG3 (YTA10) genes of Saccharomyces cerevisiae code for homologous mitochondrial proteins that belong to the recently described AAA protein-family [Kunau et al. (1993) Biochimie 75,209-224]. Mutations in either gene have been shown to induce a respiratory defect. In the case of rca1 mutants this phenotype has been ascribed to defective assembly of cytochrome oxidase and ubiquinol-cytochrome c reductase. In the present study we show that the respiratory defect of afg3 mutants, like that of rca1 mutants, is also caused by an arrest in assembly of cytochrome oxidase and ubiquinol-cytochrome c reductase. In addition to the absence of the respiratory complexes, rca1 and afg3 mutants exhibit reduced mitochondrial ATPase activity. As a first step to an understanding of the biochemical basis for the ATPase defect we have examined the assembly of the F1 and F0 constituents of the ATPase complex. We present evidence that the ATPase lesion stems at least in part from the failure of rca1 and afg3 mutants to assemble F1. Although the mutants also display lower steady-state concentrations of some F0 subunits, this could be a secondary effect of defective F1 assembly.
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Enhanced hydrogen storage properties of MgH2 co-catalyzed with K2NiF6 and CNTs.
Sulaiman, N N; Ismail, M
2016-12-06
The composite of MgH 2 /K 2 NiF 6 /carbon nanotubes (CNTs) is prepared by ball milling, and its hydrogenation properties are studied for the first time. MgH 2 co-catalyzed with K 2 NiF 6 and CNTs exhibited an improvement in the onset dehydrogenation temperature and isothermal de/rehydrogenation kinetics compared with the MgH 2 -K 2 NiF 6 composite. The onset dehydrogenation temperature of MgH 2 doped with 10 wt% K 2 NiF 6 and 5 wt% CNTs is 245 °C, which demonstrated a reduction of 25 °C compared with the MgH 2 + 10 wt% K 2 NiF 6 composite. In terms of rehydrogenation kinetics, MgH 2 doped with 10 wt% K 2 NiF 6 and 5 wt% CNTs samples absorbed 3.4 wt% of hydrogen in 1 min at 320 °C, whereas the MgH 2 + 10 wt% K 2 NiF 6 sample absorbed 2.6 wt% of hydrogen under the same conditions. For dehydrogenation kinetics at 320 °C, the MgH 2 + 10 wt% K 2 NiF 6 + 5 wt% CNTs sample released 3.3 wt% hydrogen after 5 min of dehydrogenation. By contrast, MgH 2 doped with 10 wt% K 2 NiF 6 released 3.0 wt% hydrogen in the same time period. The apparent activation energy, E a , for the dehydrogenation of MgH 2 doped with 10 wt% K 2 NiF 6 reduced from 100.0 kJ mol -1 to 70.0 kJ mol -1 after MgH 2 was co-doped with 10 wt% K 2 NiF 6 and 5 wt% CNTs. Based on the experimental results, the hydrogen storage properties of the MgH 2 /K 2 NiF 6 /CNTs composite is enhanced because of the catalytic effects of the active species of KF, KH and Mg 2 Ni that are formed in situ during dehydrogenation, as well as the unique structure of CNTs.
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'CM 88' - A multiple disease resistant chickpea mutant variety
International Nuclear Information System (INIS)
Haq, M.A.; Hassan, Mahmudul; Sadiq, M.
2001-01-01
Full text: Chickpea is the most important grain legume crop of Pakistan. Ascochyta blight (Ascochyta rabiei) and Fusarium wilt (Fusarium oxysporum F. sp cicer) are most serious diseases, having the potential to devastate a crop. A multiple disease resistant and high yielding mutant CM 88 has been developed through 100 Gy gamma irradiation treatment of variety 'C 727'. This was once a widely grown and popular variety, which lost its resistance to Ascochyta and was replaced. The selection of mutants was performed in the M2 generation grown in the Ascochyta blight nursery and sixteen mutants were selected. In the subsequent generations CM 88 proved resistant to both Ascochyta blight and Fusarium wilt, and exhibited superiority in agronomic characteristics. CM 88 was also tested for many years in the various yield trials on research stations and farmers fields throughout the country. In these trials it out yielded both the parent and standard varieties. The mutant CM 88 has been approved by the Punjab Seed Council on 27 October 1994 for general cultivation in the Punjab Province, especially the Thal area which accounts for more than 70% of the area under chickpea cultivation. (author)
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2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Lecithin. 184.1400 Section 184.1400 Food and Drugs... Substances Affirmed as GRAS § 184.1400 Lecithin. (a) Commercial lecithin is a naturally occurring mixture of... isolated as a gum following hydration of solvent-extracted soy, safflower, or corn oils. Lecithin is...
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2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Nitrogen. 184.1540 Section 184.1540 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1540 Nitrogen. (a) Nitrogen (empirical formula N2, CAS Reg. No. 7727-37-9...
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2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sorbitol. 184.1835 Section 184.1835 Food and Drugs... Substances Affirmed as GRAS § 184.1835 Sorbitol. (a) Sorbitol is the chemical 1,2,3,4,5,6-hexanehexol.... Sorbitol is produced by the electrolytic reduction, or the transition metal catalytic hydrogenation of...
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Lewis, R J; Krulwich, T A; Reynafarje, B; Lehninger, A L
1983-02-25
Obligately alkalophilic Bacillus firmus RAB had a higher molar growth yield on L-malate (Ymal = 38 mg, dry weight/mmol of L-malate) than its non-alkalophilic mutant derivative, strain RABN (Ymal = 12 mg, dry weight/mmol of L-malate). Measurements of respiration dependent proton translocation by the two strains in the presence of K+ and valinomycin showed that the alkalophile also has much higher H+/O stoichiometries (at pH 9.0) than does the mutant (at pH 7.0). H+/O ratios for B. firmus RAB at pH 9.0 were as high as 13, with a frequently observed value of 9. These high values were observed in the first phase of a set of biphasic curves for both oxygen consumption and proton ejection. At pH 7.0, both the wild type and the mutant exhibited H+/O ratios near 4 in a single phase of oxygen consumption and proton ejection. The results are consistent with suggestions that the alkalophilic respiratory chain is especially well adapted for effective energy transduction at alkaline but not neutral pH.
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Du, Jing; Huang, Xiao; Sun, Shengfang; Wang, Chunxue; Lebioda, Lukasz; Dawson, John H
2011-09-27
Dehaloperoxidase (DHP), discovered in the marine terebellid polychaete Amphitrite ornata, is the first heme-containing globin with a peroxidase activity. The sequence and crystal structure of DHP argue that it evolved from an ancient O(2) transport and storage globin. Thus, DHP retains an oxygen carrier function but also has the ability to degrade halophenol toxicants in its living environment. Sperm whale myoglobin (Mb) in the ferric state has a peroxidase activity ∼10 times lower than that of DHP. The catalytic activity enhancement observed in DHP appears to have been generated mainly by subtle changes in the positions of the proximal and distal histidine residues that appeared during DHP evolution. Herein, we report investigations into the mechanism of action of DHP derived from examination of "peroxidase-like" Mb mutants and "Mb-like" DHP mutants. The dehalogenation ability of wild-type Mb is augmented in the peroxidase-like Mb mutants (F43H/H64L, G65T, and G65I Mb) but attenuated in the Mb-like T56G DHP variant. X-ray crystallographic data show that the distal His residues in G65T Mb and G65I are positioned ∼0.3 and ∼0.8 Å, respectively, farther from the heme iron compared to that in the wild-type protein. The H93K/T95H double mutant Mb with the proximal His shifted to the "DHP-like" position has an increased peroxidase activity. In addition, a better dehaloperoxidase (M86E DHP) was generated by introducing a negative charge near His89 to enhance the imidazolate character of the proximal His. Finally, only minimal differences in dehalogenation activities are seen among the exogenous ligand-free DHP, the acetate-bound DHP, and the distal site blocker L100F DHP mutant. Thus, we conclude that binding of halophenols in the internal binding site (i.e., distal cavity) is not essential for catalysis. This work provides a foundation for a new structure-function paradigm for peroxidases and for the molecular evolution of the dual-function enzyme DHP.
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Energy Technology Data Exchange (ETDEWEB)
Du, Jing; Huang, Xiao; Sun, Shengfang; Wang, Chunxue; Lebioda, Lukasz; Dawson, John H. (SC)
2012-05-14
Dehaloperoxidase (DHP), discovered in the marine terebellid polychaete Amphitrite ornata, is the first heme-containing globin with a peroxidase activity. The sequence and crystal structure of DHP argue that it evolved from an ancient O{sub 2} transport and storage globin. Thus, DHP retains an oxygen carrier function but also has the ability to degrade halophenol toxicants in its living environment. Sperm whale myoglobin (Mb) in the ferric state has a peroxidase activity {approx}10 times lower than that of DHP. The catalytic activity enhancement observed in DHP appears to have been generated mainly by subtle changes in the positions of the proximal and distal histidine residues that appeared during DHP evolution. Herein, we report investigations into the mechanism of action of DHP derived from examination of 'peroxidase-like' Mb mutants and 'Mb-like' DHP mutants. The dehalogenation ability of wild-type Mb is augmented in the peroxidase-like Mb mutants (F43H/H64L, G65T, and G65I Mb) but attenuated in the Mb-like T56G DHP variant. X-ray crystallographic data show that the distal His residues in G65T Mb and G65I are positioned {approx}0.3 and {approx}0.8 {angstrom}, respectively, farther from the heme iron compared to that in the wild-type protein. The H93K/T95H double mutant Mb with the proximal His shifted to the 'DHP-like' position has an increased peroxidase activity. In addition, a better dehaloperoxidase (M86E DHP) was generated by introducing a negative charge near His89 to enhance the imidazolate character of the proximal His. Finally, only minimal differences in dehalogenation activities are seen among the exogenous ligand-free DHP, the acetate-bound DHP, and the distal site blocker L100F DHP mutant. Thus, we conclude that binding of halophenols in the internal binding site (i.e., distal cavity) is not essential for catalysis. This work provides a foundation for a new structure-function paradigm for peroxidases and for the
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2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Urea. 184.1923 Section 184.1923 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1923 Urea. (a) Urea (CO(NH2)2, CAS Reg. No. 57-13-6) is the diamide of...
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2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Whey. 184.1979 Section 184.1979 Food and Drugs FOOD... Substances Affirmed as GRAS § 184.1979 Whey. (a)(1) Whey. Whey is the liquid substance obtained by separating the coagulum from milk, cream, or skim milk in cheesemaking. Whey obtained from a procedure, in which...
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International Nuclear Information System (INIS)
Zapparoli, Giada V; Jorissen, Robert N; Hewitt, Chelsee A; McBean, Michelle; Westerman, David A; Dobrovic, Alexander
2013-01-01
The JAK2 V617F mutation is the most frequent somatic change in myeloproliferative neoplasms, making it an important tumour-specific marker for diagnostic purposes and for the detection of minimal residual disease. Sensitive quantitative assays are required for both applications, particularly for the monitoring of minimal residual disease, which requires not only high sensitivity but also very high specificity. We developed a highly sensitive probe-free quantitative mutant-allele detection method, Quantitative Threefold Allele-Specific PCR (QuanTAS-PCR), that is performed in a closed-tube system, thus eliminating the manipulation of PCR products. QuantTAS-PCR uses a threefold approach to ensure allele-specific amplification of the mutant sequence: (i) a mutant allele-specific primer, (ii) a 3′dideoxy blocker to suppress false-positive amplification from the wild-type template and (iii) a PCR specificity enhancer, also to suppress false-positive amplification from the wild-type template. Mutant alleles were quantified relative to exon 9 of JAK2. We showed that the addition of the 3′dideoxy blocker suppressed but did not eliminate false-positive amplification from the wild-type template. However, the addition of the PCR specificity enhancer near eliminated false-positive amplification from the wild-type allele. Further discrimination between true and false positives was enabled by using the quantification cycle (Cq) value of a single mutant template as a cut-off point, thus enabling robust distinction between true and false positives. As 10,000 JAK2 templates were used per replicate, the assay had a sensitivity of 1/10 -4 per replicate. Greater sensitivity could be reached by increasing the number of replicates analysed. Variation in replicates when low mutant-allele templates were present necessitated the use of a statistics-based approach to estimate the load of mutant JAK2 copies. QuanTAS-PCR showed comparable quantitative results when validated against a
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Differential uptake of [18F]FET and [3H]L-methionine in focal cortical ischemia
International Nuclear Information System (INIS)
Salber, Dagmar; Stoffels, Gabriele; Pauleit, Dirk; Reifenberger, Guido; Sabel, Michael; Shah, Nadim Jon; Hamacher, Kurt; Coenen, Heinz H.; Langen, Karl-Josef
2006-01-01
]MET, may exhibit significant uptake in the periphery of cortical infarctions, which has to be considered in the differential diagnosis of unknown brain lesions. There are discrepancies between [ 18 F]FET and [ 3 H]MET uptake in the area of infarctions that appear to be caused by the preferential uptake of [ 18 F]FET in reactive astrocytes versus the preferential uptake of [ 3 H]MET in macrophages
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International Nuclear Information System (INIS)
Ragab, A.I.; Boshra, S.A.; Mehany, A.L.; Darwish, A.A.; Kharrab, M.M.
2008-01-01
This study was conducted to assess 23 induced mutants and two parental varieties Giza1 and Giza2 in the three generations (M3, M4, and M5) for seed quality traits, and susceptibility to two insects i.e C. chinnensis and H.armigera. The obtained results exhibited highly significant decrease for alkaloid content of mutants 20 and 23 as compared with the two local varieties. Most of mutants and Giza 2 showed marked increase for protein content as compared with Giza1, however, the increase did not reach the level of significance for the most mutants as compared with Giza2 in the three generations. Except of M4 generation. marked resistance for infestations with C.chinensis and H.armigera was obtained for mutants 1, 5 and 11 in the three generations. However, for total infestation with the two insects, resistance was obtained in mutants 4 and 10. Except of mutant lines 1, 5 and 11, all mutants showed higher loss percentage than that of the local varieties
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Low-lying states of 184W and 184Os nuclei
International Nuclear Information System (INIS)
Sharrad, F.I.; Abdullah, Hewa Y.; Al-Dahan, N.; Umran, N.M.; Okhunov, A.A.; Abu Kassim, H.
2013-01-01
The energy levels, transition energy, B(E2) values, intrinsic quadrupole moment Q 0 and potential energy surface for even-even 184 W and 184 Os nuclei were calculated using IBM-1. The predicted energy levels, transition energy, B(E2) values and intrinsic quadrupole moment Q 0 results are reasonably consistent with the experimental data. A contour plot of the potential energy surfaces shows that two interesting nuclei are deformed and have rotational characters. (authors)
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ALS mutant SOD1 interacts with G3BP1 and affects stress granule dynamics.
Gal, Jozsef; Kuang, Lisha; Barnett, Kelly R; Zhu, Brian Z; Shissler, Susannah C; Korotkov, Konstantin V; Hayward, Lawrence J; Kasarskis, Edward J; Zhu, Haining
2016-10-01
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease. Mutations in Cu/Zn superoxide dismutase (SOD1) are responsible for approximately 20 % of the familial ALS cases. ALS-causing SOD1 mutants display a gain-of-toxicity phenotype, but the nature of this toxicity is still not fully understood. The Ras GTPase-activating protein-binding protein G3BP1 plays a critical role in stress granule dynamics. Alterations in the dynamics of stress granules have been reported in several other forms of ALS unrelated to SOD1. To our surprise, the mutant G93A SOD1 transgenic mice exhibited pathological cytoplasmic inclusions that co-localized with G3BP1-positive granules in spinal cord motor neurons. The co-localization was also observed in fibroblast cells derived from familial ALS patient carrying SOD1 mutation L144F. Mutant SOD1, unlike wild-type SOD1, interacted with G3BP1 in an RNA-independent manner. Moreover, the interaction is specific for G3BP1 since mutant SOD1 showed little interaction with four other RNA-binding proteins implicated in ALS. The RNA-binding RRM domain of G3BP1 and two particular phenylalanine residues (F380 and F382) are critical for this interaction. Mutant SOD1 delayed the formation of G3BP1- and TIA1-positive stress granules in response to hyperosmolar shock and arsenite treatment in N2A cells. In summary, the aberrant mutant SOD1-G3BP1 interaction affects stress granule dynamics, suggesting a potential link between pathogenic SOD1 mutations and RNA metabolism alterations in ALS.
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Fang, Qing; Giordimaina, Alicia M; Dolan, David F; Camper, Sally A; Mustapha, Mirna
2012-04-01
Hypothyroidism is a cause of genetic and environmentally induced deafness. The sensitivity of cochlear development and function to thyroid hormone (TH) mandates understanding TH action in this sensory organ. Prop1(df) and Pou1f1(dw) mutant mice carry mutations in different pituitary transcription factors, each resulting in pituitary thyrotropin deficiency. Despite the same lack of detectable serum TH, these mutants have very different hearing abilities: Prop1(df) mutants are mildly affected, while Pou1f1(dw) mutants are completely deaf. Genetic studies show that this difference is attributable to the genetic backgrounds. Using embryo transfer, we discovered that factors intrinsic to the fetus are the major contributor to this difference, not maternal effects. We analyzed Prop1(df) mutants to identify processes in cochlear development that are disrupted in other hypothyroid animal models but protected in Prop1(df) mutants by the genetic background. The development of outer hair cell (OHC) function is delayed, but Prestin and KCNQ4 immunostaining appear normal in mature Prop1(df) mutants. The endocochlear potential and KCNJ10 immunostaining in the stria vascularis are indistinguishable from wild type, and no differences in neurofilament or synaptophysin staining are evident in Prop1(df) mutants. The synaptic vesicle protein otoferlin normally shifts expression from OHC to IHC as temporary afferent fibers beneath the OHC regress postnatally. Prop1(df) mutants exhibit persistent, abnormal expression of otoferlin in apical OHC, suggesting delayed maturation of synaptic function. Thus, the genetic background of Prop1(df) mutants is remarkably protective for most functions affected in other hypothyroid mice. The Prop1(df) mutant is an attractive model for identifying the genes that protect against deafness.
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Hydrogen permeation behavior through F82H at high temperature
Energy Technology Data Exchange (ETDEWEB)
Matsuda, S.; Katayama, K.; Shimozori, M.; Fukada, S. [Interdisciplinary Graduate School of Engineering Science, Kyushu University, Kyushu (Japan); Ushida, H. [Energy Science and Engineering, Faculty of Engineering, Kyushu University, Kyushu (Japan); Nishikawa, M. [Malaysia-Japan International Institute of Technology, UTM, Kuala Lumpur (Malaysia)
2015-03-15
F82H is a primary candidate of structural material and coolant pipe material in a blanket of a fusion reactor. Understanding tritium permeation behavior through F82H is important. In a normal operation of a fusion reactor, the temperature of F82H will be controlled below 550 C. degrees because it is considered that F82H can be used up to 30,000 hours at 550 C. degrees. However, it is necessary to assume the situation where F82H is heated over 550 C. degrees in a severe accident. In this study, hydrogen permeation behavior through F82H was investigated in the temperature range from 500 to 800 C. degrees. In some cases, water vapor was added in a sample gas to investigate an effect of water vapor on hydrogen permeation. The permeability of hydrogen in the temperature range from 500 to 700 C. degrees agreed well with the permeability reported by E. Serra et al. The degradation of the permeability by water vapor was not observed. After the hydrogen permeation reached in a steady state at 700 C. degrees, the F82H sample was heated to 800 C. degrees. The permeability of hydrogen through F82H sample which was once heated up to 800 C. degrees was lower than that of the original one. (authors)
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National Research Council Canada - National Science Library
Scott, Maria
1999-01-01
...:H21 to human colonic epithelial cells. Transposon mutagenesis of B2F1 was accomplished with the mini-Tn5phoACm mobile element and a mutant bank of B2F1 colonies that carried putative in-frame PhoA-positive transposon insertions was isolated...
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46 CFR 184.210 - Heating equipment.
2010-10-01
... 46 Shipping 7 2010-10-01 2010-10-01 false Heating equipment. 184.210 Section 184.210 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) VESSEL CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Cooking and Heating § 184.210 Heating equipment...
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Structure of the Y94F mutant of Escherichia coli thymidylate synthase
International Nuclear Information System (INIS)
Roberts, Sue A.; Hyatt, David C.; Honts, Jerry E.; Changchien, Liming; Maley, Gladys F.; Maley, Frank; Montfort, William R.
2006-01-01
Mutation of Tyr94 of E. coli thymidylate synthase to phenylalanine leads to a protein with k cat reduced by a factor of 400. The Y94F structure is essentially identical to the wild-type structure, which is consistent with a catalytic role for the phenolic OH. Tyr94 of Escherichia coli thymidylate synthase is thought to be involved, either directly or by activation of a water molecule, in the abstraction of a proton from C5 of the 2′-deoxyuridine 5′-monophosphate (dUMP) substrate. Mutation of Tyr94 leads to a 400-fold loss in catalytic activity. The structure of the Y94F mutant has been determined in the native state and as a ternary complex with thymidine 5′-monophosphate (dTMP) and 10-propargyl 5,8-dideazafolate (PDDF). There are no structural changes ascribable to the mutation other than loss of a water molecule hydrogen bonded to the tyrosine OH, which is consistent with a catalytic role for the phenolic OH
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Structure of the Y94F mutant of Escherichia coli thymidylate synthase
Energy Technology Data Exchange (ETDEWEB)
Roberts, Sue A.; Hyatt, David C. [Department of Biochemistry and Molecular Biophysics, University of Arizona, Tucson, AZ 85721 (United States); Honts, Jerry E. [Department of Biology, Drake University, Des Moines, IA 50311 (United States); Changchien, Liming; Maley, Gladys F.; Maley, Frank [Wadsworth Center, New York State Department of Health, Albany, NY 12201-0509 (United States); Montfort, William R., E-mail: montfort@email.arizona.edu [Department of Biochemistry and Molecular Biophysics, University of Arizona, Tucson, AZ 85721 (United States)
2006-09-01
Mutation of Tyr94 of E. coli thymidylate synthase to phenylalanine leads to a protein with k{sub cat} reduced by a factor of 400. The Y94F structure is essentially identical to the wild-type structure, which is consistent with a catalytic role for the phenolic OH. Tyr94 of Escherichia coli thymidylate synthase is thought to be involved, either directly or by activation of a water molecule, in the abstraction of a proton from C5 of the 2′-deoxyuridine 5′-monophosphate (dUMP) substrate. Mutation of Tyr94 leads to a 400-fold loss in catalytic activity. The structure of the Y94F mutant has been determined in the native state and as a ternary complex with thymidine 5′-monophosphate (dTMP) and 10-propargyl 5,8-dideazafolate (PDDF). There are no structural changes ascribable to the mutation other than loss of a water molecule hydrogen bonded to the tyrosine OH, which is consistent with a catalytic role for the phenolic OH.
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High linolenic acid mutant in soybean induced by X-ray irradiation
Energy Technology Data Exchange (ETDEWEB)
Takagi, Y. [Saga Univ. (Japan); Hossain, A. B.M.M.; Yanagita, T.; Kusaba, S.
1989-12-15
Soybean [Glycine max (L.) Merr. cv. Bay] seeds were irradiated with X-rays (25kR) and the M{sub 2} progeny was screened for changes in the fatty acid composition of seed oil. X-ray irradiation remarkably increased the variability of the fatty acid composition in the oil of the Bay cultivar. A mutant in which linolenic acid accounted for 18.4 per cent of the total oil cornpared with 9.4 per cent in the Bay cultivar was identified among 2006 M{sub 2} plants. The M{sub 3} generation of the mutant also showed a linolenic acid content approximately two times higher than that of the original variety.
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High linolenic acid mutant in soybean induced by X-ray irradiation
International Nuclear Information System (INIS)
Takagi, Y.; Hossain, A.B.M.M.; Yanagita, T.; Kusaba, S.
1989-01-01
Soybean [Glycine max (L.) Merr. cv. Bay] seeds were irradiated with X-rays (25kR) and the M 2 progeny was screened for changes in the fatty acid composition of seed oil. X-ray irradiation remarkably increased the variability of the fatty acid composition in the oil of the Bay cultivar. A mutant in which linolenic acid accounted for 18.4 per cent of the total oil cornpared with 9.4 per cent in the Bay cultivar was identified among 2006 M 2 plants. The M 3 generation of the mutant also showed a linolenic acid content approximately two times higher than that of the original variety
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Genetic Segregation Analysis of a Rapeseed Dwarf Mutant
International Nuclear Information System (INIS)
Xiang, G.; Yu, S.; Zhang, T.; Zhao, J.; Lei, S.; Du, C.
2016-01-01
Dwarf resources in Brassica napus are very important for developing high-yield cultivars through dwarf-type and lodging-resistant breeding. However, few dwarf varieties have been available for this species. Here, we reported a new rapeseed dwarf mutant GRC1157, which exhibits obvious phenotypic variations on dwarf. Six generations (P /sub 1/, P/sub 1/, F/sub 1/, F/sub 1/, B/sub 1/, and B/sub 1/) were produced from a cross between dwarf mutant GRC1157 and an elite tall-type line XR16 to analyze genetic inheritances of plant height (PH), numbers of the 1st valid branch (VBN), main inflorescence length (MIL), pod numbers per main inflorescence (MPN), pod length (PL) and seed numbers per pod (PSN) using the mixed major gene plus polygene inheritance model. The genetic analysis shows different traits were controlled by different inheritance models: PH and PL by two pairs of additive-dominant-epistatic major genes plus additive-dominant-epistatic polygenes, MPN and PSN by two-pair additive-dominant-epistatic major genes plus additive-dominant polygenes, MIL by two-pair additive-dominant-epistatic major genes and VBN by one-pair additive-dominant major genes plus additive-dominant-epistatic polygenes. Furthermore, positive correlations between PH and some other traits were observed, suggesting that some traits may be co-regulated by several linkage or same loci/genes. In addition, high heritability (40.35-93.7 percent) were found for five traits (except VBN) in different segregating generations, indicating these traits were mainly affected by hereditary factors and suitable for early artificial selection. In sum, the dwarf mutant GRC1157 can serve as a valuable resource for rapeseed dwarf breeding and the genetic analysis in this study provided a foundation for further mapping and cloning dwarf genes in mutant GRC1157. (author)
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Klinsupa, Worawan; Phansiri, Salak; Thongpradis, Panida; Yongsmith, Busaba; Pothiratana, Chetsada
2016-01-10
To breed industrially useful strains of a slow-growing, yellow pigment producing strain of Monascus sp., protoplasts of Monascus purpureus yellow mutant (ade(-)) and rapid-growing M. purpureus white mutant (prototroph) were fused and fusants were selected on minimal medium (MM). Preliminary conventional protoplast fusion of the two strains was performed and the result showed that only white colonies were detected on MM. It was not able to differentiate the fusants from the white parental prototroph. To solve this problem, the white parental prototroph was thus pretreated with 20mM iodoacetamide (IOA) for cytoplasm inactivation and subsequently taken into protoplast fusion with slow-growing Monascus yellow mutant. Under this development technique, only the fusants, with viable cytoplasm from Monascus yellow mutant (ade(-)), could thus grow on MM, whereas neither IOA pretreated white parental prototroph nor yellow auxotroph (ade(-)) could survive. Fifty-three fusants isolated from yellow colonies obtained through this developed technique were subsequently inoculated on complete medium (MY agar). Fifteen distinguished yellow colonies from their parental yellow mutant were then selected for biochemical, morphological and fermentative properties in cassava starch and soybean flour (SS) broth. Finally, three most stable fusants (F7, F10 and F43) were then selected and compared in rice solid culture. Enhancement of yellow pigment production over the parental yellow auxotroph was found in F7 and F10, while enhanced glucoamylase activity was found in F43. The formation of fusants was further confirmed by monacolin K content, which was intermediate between the two parents (monacolin K-producing yellow auxotroph and non-monacolin K producing white prototroph). Copyright © 2015 Elsevier B.V. All rights reserved.
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21 CFR 184.1631 - Potassium hydroxide.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Potassium hydroxide. 184.1631 Section 184.1631 Food... Specific Substances Affirmed as GRAS § 184.1631 Potassium hydroxide. (a) Potassium hydroxide (KOH, CAS Reg... pellets, flakes, sticks, lumps, and powders. Potassium hydroxide is obtained commercially from the...
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46 CFR 184.220 - Cooking equipment.
2010-10-01
... 46 Shipping 7 2010-10-01 2010-10-01 false Cooking equipment. 184.220 Section 184.220 Shipping...) VESSEL CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Cooking and Heating § 184.220 Cooking equipment. (a) Doors on a cooking appliance must be provided with hinges and locking devices to prevent...
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Structure of doubly-odd nucleus 184Ir from the decay of 184Pt
International Nuclear Information System (INIS)
Ben Braham, A.; Bourgeois, C.; Kilcher, P.; Roussiere, B.; Sauvage, J.; Kreiner, A.J.; Porquet, M.G.
1987-01-01
States of 184 Ir populated through the β + /EC decay of 184 Pt, were studied using mass-separated sources and on-line γ-ray and e - spectroscopy techniques. A rather complete low-spin level scheme is established. Negative and positive parity structures are interpreted within the two-quasiparticle-plus-rotor model
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21 CFR 184.1622 - Potassium chloride.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Potassium chloride. 184.1622 Section 184.1622 Food... Specific Substances Affirmed as GRAS § 184.1622 Potassium chloride. (a) Potassium chloride (KCl, CAS Reg... levels not to exceed current good manufacturing practice. Potassium chloride may be used in infant...
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Xia, Zhen Wei; Cui, Wen Jun; Zhou, Wen Pu; Zhang, Xue Hong; Shen, Qing Xiang; Li, Yun Zhu; Yu, Shan Chang
2004-10-01
Human Heme Oxygenase-1 (hHO-1) is the rate-limiting enzyme in the catabolism reaction of heme, which directly regulates the concentration of bilirubin in human body. The mutant structure was simulated by Swiss-pdbviewer procedure, which showed that the structure of active pocket was changed distinctly after Ala25 substituted for His25 in active domain, but the mutated enzyme still binded with heme. On the basis of the results, the expression vectors, pBHO-1 and pBHO-1(M), were constructed, induced by IPTG and expressed in E. coli DH5alpha strain. The expression products were purified with 30%-60% saturation (NH4)2SO4 and Q-Sepharose Fast Flow column chromatography. The concentration of hHO-1 in 30%-60% saturation (NH4)2SO4 components and in fractions through twice column chromatography was 3.6-fold and 30-fold higher than that in initial product, respectively. The activity of wild hHO-1 (whHO-1) and mutant hHO-1 (deltahHO-1) showed that the activity of deltahHO-1 was reduced 91.21% compared with that of whHO-1. The study shows that His25 is of importance for the mechanism of hHO-1, and provides the possibility for effectively regulating the activity to exert biological function.
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21 CFR 184.1643 - Potassium sulfate.
2010-04-01
... hydroxide or potassium carbonate. (b) The ingredient meets the specifications of the “Food Chemicals Codex... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Potassium sulfate. 184.1643 Section 184.1643 Food... Specific Substances Affirmed as GRAS § 184.1643 Potassium sulfate. (a) Potassium sulfate (K2SO4, CAS Reg...
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21 CFR 184.1702 - Sheanut oil.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sheanut oil. 184.1702 Section 184.1702 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1702 Sheanut oil. (a) Sheanut oil is produced from sheanuts derived from...
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21 CFR 184.1555 - Rapeseed oil.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Rapeseed oil. 184.1555 Section 184.1555 Food and... Substances Affirmed as GRAS § 184.1555 Rapeseed oil. (a) Fully hydrogenated rapeseed oil. (1) Fully hydrogenated rapeseed oil is a mixture of triglycerides in which the fatty acid composition is a mixture of...
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2010-10-01
... 46 Shipping 7 2010-10-01 2010-10-01 false Gas systems. 184.240 Section 184.240 Shipping COAST... CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Cooking and Heating § 184.240 Gas systems. Cooking systems... requirements: (a) The design, installation and testing of each LPG system must meet ABYC A-1, “Marine Liquefied...
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21 CFR 184.1848 - Starter distillate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Starter distillate. 184.1848 Section 184.1848 Food... Specific Substances Affirmed as GRAS § 184.1848 Starter distillate. (a) Starter distillate (butter starter distillate) is a steam distillate of the culture of any or all of the following species of bacteria grown on...
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21 CFR 184.1351 - Gum tragacanth.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Gum tragacanth. 184.1351 Section 184.1351 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1351 Gum tragacanth. (a) Gum tragacanth is the exudate from one of several...
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International Nuclear Information System (INIS)
Strobel, G.A.; Steiner, G.W.; Byther, R.
1975-01-01
Three mutants selected from a population of sugarcane clone H54-775 that had been irradiated with 3 kR γ-radiation all lacked toxin-binding protein activity. This activity previously had been shown to be essential for eye spot disease susceptibility and was demonstrated in the susceptible parent clone H54-775. In one mutant, the biochemical, immunochemical, and electrophoretic mobilities of the toxin-binding protein were all modified
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21 CFR 184.1978 - Carnauba wax.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Carnauba wax. 184.1978 Section 184.1978 Food and... Substances Affirmed as GRAS § 184.1978 Carnauba wax. (a) Carnauba wax (CAS Reg. No. 008-015-869) is obtained from the leaves and buds of the Brazilian wax palm Copernicia cerifera Martius. The wax is hard...
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21 CFR 184.1431 - Magnesium oxide.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Magnesium oxide. 184.1431 Section 184.1431 Food and... Substances Affirmed as GRAS § 184.1431 Magnesium oxide. (a) Magnesium oxide (MgO, CAS Reg. No. 1309-48-4... powder (light) or a relatively dense white powder (heavy) by heating magnesium hydroxide or carbonate...
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21 CFR 184.1322 - Wheat gluten.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Wheat gluten. 184.1322 Section 184.1322 Food and... Substances Affirmed as GRAS § 184.1322 Wheat gluten. (a) Wheat gluten (CAS Reg. No. 8002-80-0) is the principal protein component of wheat and consists mainly of gliadin and glutenin. Wheat gluten is obtained...
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21 CFR 184.1321 - Corn gluten.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Corn gluten. 184.1321 Section 184.1321 Food and... Substances Affirmed as GRAS § 184.1321 Corn gluten. (a) Corn gluten (CAS Reg. No. 66071-96-3), also known as corn gluten meal, is the principal protein component of corn endosperm. It consists mainly of zein and...
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2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Corn syrup. 184.1865 Section 184.1865 Food and... Substances Affirmed as GRAS § 184.1865 Corn syrup. (a) Corn syrup, commonly called “glucose sirup” or “glucose syrup,” is obtained by partial hydrolysis of corn starch with safe and suitable acids or enzymes...
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21 CFR 184.1859 - Invert sugar.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Invert sugar. 184.1859 Section 184.1859 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1859 Invert sugar. (a) Invert sugar (CAS Reg. No. 8013-17-0) is an aqueous...
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21 CFR 184.1240 - Carbon dioxide.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Carbon dioxide. 184.1240 Section 184.1240 Food and... Substances Affirmed as GRAS § 184.1240 Carbon dioxide. (a) Carbon dioxide (empirical formula CO2, CAS Reg. No.... The solid form, dry ice, sublimes under atmospheric pressure at a temperature of −78.5 °C. Carbon...
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21 CFR 184.1472 - Menhaden oil.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Menhaden oil. 184.1472 Section 184.1472 Food and... Substances Affirmed as GRAS § 184.1472 Menhaden oil. (a) Menhaden oil. (1) Menhaden oil is prepared from fish of the genus Brevoortia, commonly known as menhaden, by cooking and pressing. The resulting crude oil...
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21 CFR 184.1138 - Ammonium chloride.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Ammonium chloride. 184.1138 Section 184.1138 Food... Specific Substances Affirmed as GRAS § 184.1138 Ammonium chloride. (a) Ammonium chloride (NH4Cl, CAS Reg. No. 12125-02-9) is produced by the reaction of sodium chloride and an ammonium salt in solution. The...
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21 CFR 184.1297 - Ferric chloride.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Ferric chloride. 184.1297 Section 184.1297 Food and... Substances Affirmed as GRAS § 184.1297 Ferric chloride. (a) Ferric chloride (iron (III) chloride, FeC13, CAS Reg. No. 7705-08-0) may be prepared from iron and chlorine or from ferric oxide and hydrogen chloride...
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21 CFR 184.1545 - Nitrous oxide.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Nitrous oxide. 184.1545 Section 184.1545 Food and... Substances Affirmed as GRAS § 184.1545 Nitrous oxide. (a) Nitrous oxide (empirical formula N2O, CAS Reg. No.... Nitrous oxide is manufactured by the thermal decomposition of ammonium nitrate. Higher oxides of nitrogen...
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21 CFR 184.1742 - Sodium carbonate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sodium carbonate. 184.1742 Section 184.1742 Food... Specific Substances Affirmed as GRAS § 184.1742 Sodium carbonate. (a) Sodium carbonate (Na2CO3, CAS Reg. No... ore calcined to impure soda ash and then purified; or (3) synthesized from limestone by the Solvay...
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21 CFR 184.1415 - Animal lipase.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Animal lipase. 184.1415 Section 184.1415 Food and... Substances Affirmed as GRAS § 184.1415 Animal lipase. (a) Animal lipase (CAS Reg. No. 9001-62-1) is an enzyme preparation obtained from edible forestomach tissue of calves, kids, or lambs, or from animal pancreatic...
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Directory of Open Access Journals (Sweden)
Gian Marco Tosi
2014-01-01
Full Text Available Purpose. To evaluate the effectiveness and safety of perfluorohexyloctane (F6H8 for intraoperative flattening of the retina and of F6H8/silicone oil (SO 1000 cSt as a postoperative tamponade for inferior retinal detachment with inferior proliferative vitreoretinopathy. Methods. This is a retrospective review of 22 patients who underwent pars plana vitrectomy using F6H8 as an intraoperative tool to flatten the retina. At the end of the surgery a direct partial exchange between F6H8 and SO 1000 cSt was performed, tamponing the eye with different ratios of F6H8/SO (70/30, 60/40, 50/50, 40/30, and 30/70. Anatomical and functional results and complications were evaluated over the follow-up period (mean 22.63 months. Results. F6H8 was efficacious for intraoperative flattening of the retina. Twenty-one of the 22 patients achieved a complete retinal reattachment. Postoperative visual acuity (VA ranged from light perception to 20/70, with 72% of patients obtaining VA better than 20/400. No emulsification/inflammation was observed whatever the ratio of F6H8/SO used. With higher ratios of F6H8/SO (70/30 and 60/40 cloudiness of the tamponade was observed. A transparent mixture was present with all the other ratios. Conclusions. The surgical technique adopted is very simple and safe. The optimal F6H8/SO ratio seems to be between 50/50 and 30/70.
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21 CFR 184.1210 - Calcium oxide.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Calcium oxide. 184.1210 Section 184.1210 Food and... Substances Affirmed as GRAS § 184.1210 Calcium oxide. (a) Calcium oxide (CaO, CAS Reg. No. 1305-78-8) is also known as lime, quick lime, burnt lime, or calx. It is produced from calcium carbonate, limestone, or...
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2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Vitamin A. 184.1930 Section 184.1930 Food and Drugs... Substances Affirmed as GRAS § 184.1930 Vitamin A. (a)(1) Vitamin A (retinol; CAS Reg. No. 68-26-8) is the... odorless or have a mild fishy odor. Vitamin A is extracted from fish liver oils or produced by total...
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21 CFR 184.1890 - α-Tocopherols.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true α-Tocopherols. 184.1890 Section 184.1890 Food and... Substances Affirmed as GRAS § 184.1890 α-Tocopherols. (a) The α-tocopherols that are the subject of this GRAS affirmation regulation are limited to the following: (1) d-α-Tocopherol (CAS Reg. No. 59-02-9) is the chemical...
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2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true L-Cysteine. 184.1271 Section 184.1271 Food and... Substances Affirmed as GRAS § 184.1271 L-Cysteine. (a) L-Cysteine is the chemical L-2-amino-3... of total L-cysteine per 100 parts of flour in dough as a dough strengthener as defined in § 170.3(o...
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Abdel-Azeim, Safwat; Oliva, Romina M.; Chermak, Edrisse; De Cristofaro, Raimondo; Cavallo, Luigi
2014-01-01
Factor X (FX) is one of the major players in the blood coagulation cascade. Upon activation to FXa, it converts prothrombin to thrombin, which in turn converts fibrinogen into fibrin (blood clots). FXa deficiency causes hemostasis defects, such as intracranial bleeding, hemathrosis, and gastrointestinal blood loss. Herein, we have analyzed a pool of pathogenic mutations, located in the FXa catalytic domain and directly associated with defects in enzyme catalytic activity. Using chymotrypsinogen numbering, they correspond to D102N, T135M, V160A, G184S, and G197D. Molecular dynamics simulations were performed for 1.68 μs on the wild-type and mutated forms of FXa. Overall, our analysis shows that four of the five mutants considered, D102N, T135M, V160A, and G184S, have rigidities higher than those of the wild type, in terms of both overall protein motion and, specifically, subpocket S4 flexibility, while S1 is rather insensitive to the mutation. This acquired rigidity can clearly impact the substrate recognition of the mutants.
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Abdel-Azeim, Safwat
2014-11-11
Factor X (FX) is one of the major players in the blood coagulation cascade. Upon activation to FXa, it converts prothrombin to thrombin, which in turn converts fibrinogen into fibrin (blood clots). FXa deficiency causes hemostasis defects, such as intracranial bleeding, hemathrosis, and gastrointestinal blood loss. Herein, we have analyzed a pool of pathogenic mutations, located in the FXa catalytic domain and directly associated with defects in enzyme catalytic activity. Using chymotrypsinogen numbering, they correspond to D102N, T135M, V160A, G184S, and G197D. Molecular dynamics simulations were performed for 1.68 μs on the wild-type and mutated forms of FXa. Overall, our analysis shows that four of the five mutants considered, D102N, T135M, V160A, and G184S, have rigidities higher than those of the wild type, in terms of both overall protein motion and, specifically, subpocket S4 flexibility, while S1 is rather insensitive to the mutation. This acquired rigidity can clearly impact the substrate recognition of the mutants.
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Radiation induced mutants in cassava (Manihot esculenta Crantz)
International Nuclear Information System (INIS)
Nayar, G.G.; Rajendran, P.G.
1987-01-01
Full text: Stem cuttings and true seeds of three promising cultivars of cassava were exposed respectively to 1 to 5 kR and 10 to 50 kR acute gamma rays from a 60 Co source. Treatments of stem cuttings beyond 5 kR and seeds beyond 50 kR were lethal. One mutant each in the cultivars M4, H-165 and H-2304 was obtained from the stem irradiated populations. Another mutant was found in the seed irradiated progeny of H-2304. The mutant of M4 is characterised by light green (chlorina) leaves. The mutant of H-165 shows significantly shorter petiole (22,5 against 35.2 cm) and narrow leaf lobes, while the H-2304 mutant shows speckled leaves, branching and early flowering. The mutant found in the seed irradiated progeny of H-2304 is having yellow tuber flesh indicating the presence of carotene. The mutants may be useful in studies related to basic information as well as in practical breeding. The chlorina mutant in M4 showed slow growth and high HCN content in leaves. Late branching may be a useful trait in the traditionally non-branching clones of cassava to maintain the desirable leaf area index during high leaf fall period. Early flowering could be useful in a recombinant breeding programme. The tuber yield of the short petiole mutant in H-165 increased by 20% - 25% through closer planting. The narrow leaf lobes of this mutant permit better light penetration to lower leaves. (author)
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Study of the decay /sup 184/Hg. -->. /sup 184/Au (Tsub(1/2)=30s). [Lifetime, upper limits
Energy Technology Data Exchange (ETDEWEB)
Nettles, W G; Beraud, R; Cole, J D; Hamilton, J H; Ramayya, A V; Kawakami, H [Vanderbilt Univ., Nashville, TN (USA). Dept. of Physics; Sastry, K S.R. [Massachusetts Univ., Amherst (USA); Spejewski, E H
1978-04-01
The radioactive decay of /sup 184/Hg to /sup 184/Au was studied on line with the UNISOR isotope separator. The 184 mass chain was entered at /sup 184/Tl and /sup 184/Hg by bombarding an isotopically-enriched target of /sup 180/W with /sup 14/N ions of 177 MeV. Multiscale singles and ..gamma..-..gamma..-time coincidence studies were carried out. No previous levels have been assigned to /sup 184/Au. From the coincidence data eight levels at 156.2, 159.1, 247.7, 282.8, 295.1, 392.4, 418.0 and 412.4 keV are assigned and two tentative levels at 534.0 and 551.6 keV are proposed. By using the delayed coincidence technique, a half-life of 36+-6 nanoseconds was measured for the 156.2 keV first-excited level and upper limits of 2 nanoseconds were determined for the half-lives of levels at 159.1, 247.7, 295.1 and 392.4 keV.
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International Nuclear Information System (INIS)
Oishi, Motoi; Sumitani, Shogo; Nagasaki, Yukio; Bronich, Tatiana K.; Kabanov, Alexander V.; Boska, Michael D.
2009-01-01
The pH-responsive PEGylated nanogels composed of the cross-linked poly[2-(N,N-diethylamino)ethyl methacrylate]-co-poly(2,2,2-trifluoroethyl methacrylate) gel core showed a remarkable on-off regulation of 19 F magnetic resonance signal intensity (T 2 values) as well as signal-to-noise ratios in response to extracellular pH 6.5 of tumor environment under 19 F magnetic resonance spectroscopic imaging (MRS/I), demonstrating the utility of the PEGylated nanogels as solid tumor-specific 19 F MRI/S nanoprobes. (author)
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International Nuclear Information System (INIS)
Jayasundera, Anil C.A.; Goff, Richard J.; Li Yang; Finch, Adrian A.; Lightfoot, Philip
2010-01-01
The solvothermal syntheses and crystal structures of three indium fluorides are presented. K 5 In 3 F 14 (1) and β-(NH 4 ) 3 InF 6 (2) are variants on known inorganic structure types chiolite and cryolite, respectively, with the latter exhibiting a complex and apparently novel structural distortion. [NH 4 ] 3 [C 6 H 21 N 4 ] 2 [In 4 F 21 ] (3) represents a new hybrid composition displaying a unique trimeric metal fluoride building unit. - Graphical abstract: Solvothermal synthesis has been used to prepare three indium fluorides, including a novel hybrid material containing a unique [In 3 F 15 ] trimer templated by tren.
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Korenev, Vladimir S; Floquet, Sébastien; Marrot, Jérôme; Haouas, Mohamed; Mbomekallé, Israël-Martyr; Taulelle, Francis; Sokolov, Maxim N; Fedin, Vladimir P; Cadot, Emmanuel
2012-02-20
Reaction of the cyclic lacunary [H(7)P(8)W(48)O(184)](33-) anion (noted P(8)W(48)) with the [Mo(2)S(2)O(2)(H(2)O)(6)](2+) oxothiocation led to two compounds, namely, [K(4){Mo(4)O(4)S(4)(H(2)O)(3)(OH)(2)}(2)(WO(2))(P(8)W(48)O(184))](30-) (denoted 1) and [{Mo(4)O(4)S(4)(H(2)O)(3)(OH)(2)}(2)(P(8)W(48)O(184))](36-) (denoted 2), which were characterized in the solid state and solution. In the solid state, the structure of [K(4){Mo(4)O(4)S(4)(H(2)O)(3)(OH)(2)}(2)(WO(2))(P(8)W(48)O(184))](30-) reveals the presence of two disordered {Mo(4)O(4)S(4)(H(2)O)(3)(OH)(2)}(2+) "handles" connected on both sides of the P(8)W(48) ring. Such a disorder is consistent with the presence of two geometrical isomers where the relative disposition of the two {Mo(4)O(4)S(4)(H(2)O)(3)(OH)(2)}(2+) handles are arranged in a perpendicular or parallel mode. Such an interpretation is fully supported by (31)P and (183)W NMR solution studies. The relative stability of both geometrical isomers appears to be dependent upon the nature of the internal alkali cations, i.e., Na(+) vs K(+), and increased lability of the two {Mo(4)O(4)S(4)(H(2)O)(3)(OH)(2)}(2+) handles, compared to the oxo analogous, was clearly identified by significant broadening of the (31)P and (183)W NMR lines. Solution studies carried out by UV-vis spectroscopy showed that formation of the adduct [{Mo(4)O(4)S(4)(H(2)O)(3)(OH)(2)}(2)(P(8)W(48)O(184))](36-) occurs in the 1.5-4.7 pH range and corresponds to a fast and quantitative condensation process. Furthermore, (31)P NMR titrations in solution reveal formation of the "monohandle" derivative [{Mo(4)O(4)S(4)(H(2)O)(3)(OH)(2)}(P(8)W(48)O(184))](38-) as an intermediate prior to formation of the "bishandle" derivatives. Furthermore, the electrochemical behavior of [{Mo(4)O(4)S(4)(H(2)O)(3)(OH)(2)}(2)(P(8)W(48)O(184))](36-) was studied in aqueous medium and compared with the parent anion P(8)W(48).
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Jura, Marek; Levason, William; Reid, Gillian; Webster, Michael
2009-10-07
Two very unusual sulfonium salts, [{o-C(6)H(4)(CH(2)SMe)(2)}H][NbF(6)] and [{[9]aneS(3)}H][NbF(6)], obtained from reaction of the thioethers with NbF(5) in CH(2)Cl(2) solution, are reported and their structures described; the eight-coordinate tetrafluoro Nb(v) cation of the dithioether is obtained from the same reaction.
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Preliminary results of the round-robin testing of F82H
Energy Technology Data Exchange (ETDEWEB)
Shiba, K.; Yamanouchi, N.; Tohyama, A.
1996-10-01
Preliminary results of metallurgical, physical and mechanical properties of low activation ferritic steel F82H (IEA heat) were obtained in the round-robin test in Japan. The properties of IEA heat F82H were almost the same as the original F82H.
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International Nuclear Information System (INIS)
Cornehl, H.H.; Hornung, G.; Schwarz, H.
1996-01-01
The gas-phase reactivity of the fluorinated hydrocarbons CF 4 , CHF 3 , CH 3 F, C 2 F 6 , 1,1-C 2 H 4 F 2 , and C 6 F 6 with the lanthanide cations Ce + , Pr + , Sm + , Ho + , Tm + , and Yb + and the reactivity of C 6 H 5 F with all lanthanide cations Ln + (Ln = La-Lu, with the exception of Pm + ) have been examined by Fourier-transform ion cyclotron resonance mass spectrometry. The perfluorinated compounds tetrafluoromethane and hexafluoroethane as well as trifluoromethane do not react with any lanthanide cation. Selective activation of the strong C-F bonds in fluoromethane, 1,1-difluoroethane, hexafluorobenzene, and fluorobenzene appears as a general reaction scheme along the 4f row. Experimental evidence is given for a 'harpoon'-like mechanism for the F atom abstraction process which operates via an initial electron transfer from the lanthanide cation to the fluorinated substrate in the encounter complex Ln + RF. The most reactive lanthanides La + , Ce + , Gd + , and Tb + and also the formal closed-shell species Lu + exhibit additional C-H and C-C bond activation pathways in the reaction with fluorobenzene, namely dehydrohalogenation as well as loss of a neutral acetylene molecule. In the case of Tm + and Yb + the formation of neutral LnF 3 is observed in a multistep process via C-C coupling and charge transfer. 17 refs., 2 figs., 2 tabs
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Subunit-specific phenotypes of Salmonella typhimurium HU mutants.
Hillyard, D R; Edlund, M; Hughes, K T; Marsh, M; Higgins, N P
1990-01-01
Salmonella hupA and hupB mutants were studied to determine the reasons for the high degree of conservation in HU structure in bacteria. We found one HU-1-specific effect; the F'128 plasmid was 25-fold less stable in hupB compared with hupA or wild-type cells. F' plasmids were 120-fold more unstable in hupA hupB double mutants compared with wild-type cells, and the double mutant also had a significant alteration in plasmid DNA structure. pBR322 DNA isolated from hupA hupB strains was deficient...
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Energy Technology Data Exchange (ETDEWEB)
Gerasimenko, Andrey V.; Gaivoronskaya, Kseniya A.; Slobodyuk, Arseny B.; Didenko, Nina A. [Institute of Chemistry, Russian Academy of Sciences, Vladivostok (Russian Federation)
2017-12-04
The MgZrF{sub 6}.nH{sub 2}O (n = 5, 2 and 0) compounds were studied by the methods of X-ray diffraction and {sup 19}F, MAS {sup 19}F, and {sup 1}H NMR spectroscopy. At room temperature, the compound MgZrF{sub 6}.5H{sub 2}O has a monoclinic C-centered unit cell and is composed of isolated chains of edge-sharing ZrF{sub 8} dodecahedra reinforced with MgF{sub 2}(H{sub 2}O){sub 4} octahedra and uncoordinated H{sub 2}O molecules and characterized by a disordered system of hydrogen bonds. In the temperature range 259 to 255 K, a reversible monoclinic <-> two-domain triclinic phase transition is observed. The phase transition is accompanied with ordering of hydrogen atoms positions and the system of hydrogen bonds. The structure of MgZrF{sub 6}.2H{sub 2}O comprises a three-dimensional framework consisting of chains of edge-sharing ZrF{sub 8} dodecahedra linked to each other through MgF{sub 4}(H{sub 2}O){sub 2} octahedra. The compound MgZrF{sub 6} belongs to the NaSbF{sub 6} type and is built from regular ZrF{sub 6} and MgF{sub 6} octahedra linked into a three-dimensional framework through linear Zr-F-Mg bridges. The peaks in {sup 19}F MAS spectra were attributed to the fluorine structural positions. The motions of structural water molecules were studied by variable-temperature {sup 1}H NMR spectroscopy. (copyright 2017 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)
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Spread-F occurrences and relationships with foF2 and h'F at low- and mid-latitudes in China
Wang, Ning; Guo, Lixin; Zhao, Zhenwei; Ding, Zonghua; Lin, Leke
2018-04-01
Ionospheric irregularities are an important phenomenon in scientific studies and applications of radio-wave propagation. Spread-F echoes in ionograms are a type of high-frequency band irregularities that include frequency spread-F (FSF), range spread-F (RSF), and mixed spread-F (MSF) events. In this study, we obtained spread-F data from four ionosondes at low- and mid-latitudes near the 120°E chain in China during the 23rd solar cycle. We used these data to investigate spread-F occurrence percentages and variations with local time, season, latitude, and solar activity. The four ionosondes were located at Haikou (HK) (20°N, 110.34°E), Guangzhou (GZ) (23.14°N, 113.36°E), Beijing (BJ) (40.11°N, 116.28°E), and Changchun (CC) (43.84°N, 125.28°E). We also present possible correlations between spread-Fs and other ionospheric parameters, such as the critical frequency of the F2-layer (foF2) and the virtual height of the bottom-side F-layer (h'F). In particular, we investigated the possible threshold of the foF2 affecting the FSF and the relationship between the h'F and the RSF. The main conclusions are as follows: (a) the FSF occurrence percentages were anti-correlated with solar activity at all four sites; meanwhile, RSF occurrence rates increased with the increase in solar activity at HK, but not at the other three sites; (b) FSF occurrence rates were larger at the mid-latitudes than expected, while FSFs occurred more often after midnight; (c) the highest FSF occurrence rates mostly appeared during the summer months, while RSFs occurred mostly in the equinoctial months of 2000-2002 at HK and GZ; (d) a lower foF2 was suitable for FSF events; nevertheless, h'F and RSF occurrences satisfied the parabolic relationship; (e) the foF2 thresholds for FSFs were 15, 14, 7.6, and 7.8 MHz at HK, GZ, BJ, and CC, respectively. The h'Fs occurring between 240 and 290 km were more favorable for RSF occurrences. These results are important for understanding ionospheric
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Zhou, Yongjin; Wang, Lei; Yang, Fan; Lin, Xinping; Zhang, Sufang; Zhao, Zongbao K.
2011-01-01
NAD (NAD+) and its reduced form (NADH) are omnipresent cofactors in biological systems. However, it is difficult to determine the extremes of the cellular NAD(H) level in live cells because the NAD+ level is tightly controlled by a biosynthesis regulation mechanism. Here, we developed a strategy to determine the extreme NAD(H) levels in Escherichia coli cells that were genetically engineered to be NAD+ auxotrophic. First, we expressed the ntt4 gene encoding the NAD(H) transporter in the E. coli mutant YJE001, which had a deletion of the nadC gene responsible for NAD+ de novo biosynthesis, and we showed NTT4 conferred on the mutant strain better growth in the presence of exogenous NAD+. We then constructed the NAD+-auxotrophic mutant YJE003 by disrupting the essential gene nadE, which is responsible for the last step of NAD+ biosynthesis in cells harboring the ntt4 gene. The minimal NAD+ level was determined in M9 medium in proliferating YJE003 cells that were preloaded with NAD+, while the maximal NAD(H) level was determined by exposing the cells to high concentrations of exogenous NAD(H). Compared with supplementation of NADH, cells grew faster and had a higher intracellular NAD(H) level when NAD+ was fed. The intracellular NAD(H) level increased with the increase of exogenous NAD+ concentration, until it reached a plateau. Thus, a minimal NAD(H) level of 0.039 mM and a maximum of 8.49 mM were determined, which were 0.044× and 9.6× those of wild-type cells, respectively. Finally, the potential application of this strategy in biotechnology is briefly discussed. PMID:21742902
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F/H Effluent Treatment Facility. Preliminary engineering report
International Nuclear Information System (INIS)
1985-01-01
The Department of Energy is currently proposing to construct the F/H ETF to process wastewater from the Separations Areas and replace the existing seepage basins. Reasons for seepage basin closure are two-fold. First, nonradioactive hazardous materials routinely discharged to the seepage basins may have adversely impacted the quality of the groundwater in the vicinity of the basins. Second, amendments to the Resource Conservation and Recovery Act (RCRA) were approved in 1984, prohibiting the discharge of hazardous wastes to unlined seepage basins after November, 1988. The F/H ETF will consist of wastewater storage facilities and a treatment plant discharging treated effluent to Upper Three Runs Creek. Seepage basin use in F and H Areas wil be discontinued after startup, allowing timely closure of these basins. 3 refs
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Single-crystal structure refinement of YbF{sub 2} with a remark about YbH{sub 2}
Energy Technology Data Exchange (ETDEWEB)
Reckeweg, Olaf; DiSalvo, Francis J. [Cornell Univ., Ithaca, NY (United States). Dept. of Chemistry and Chemical Biology
2017-07-01
Transparent-yellow single crystals of YbF{sub 2} were obtained as only crystalline product from the solid-state reaction of Yb and teflon designed to yield 'Yb{sub 3}C{sub 3}F{sub 2}' in addition to some amorphous black material. The first single-crystal structure determination of YbF{sub 2} (cubic space group Fm anti 3m, CaF{sub 2}-type structure, a = 559.46(16) pm; R1 = 1.2%, wR2 = 3.2%) was the starting point to compare isostructural binary fluorides MF{sub 2} and hydrides MH{sub 2} (M = Ca, Yb, Eu, Sr and Ba) exhibiting an as-yet unexplained small volume per formula unit for YbH{sub 2}.
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International Nuclear Information System (INIS)
Saito, H.; Hayashi, Y.; Abe, T.; Kanaya, T.; Suzuki, K.
2005-01-01
Full text: Garden verbenas (Verbena hybrida) belonging to the Verbenaceae family are originated from interspecific hybridization among several species, many cultivars frequently produce seeds. Recently, a sterile mutant has been isolated in the verbena cultivar 'Coral Pink' of Temari series (Suntory Flowers Ltd., Tokyo, Japan) by mutation induction using heavy-ion beams at RIKEN Accelarator Research Facility (RARF, Saitama, Japan). We investigated pollen and ovule fertility assessed by acetocarmin staining, seed-set following controlled-pollination tests and behavior of pollen tubes in pistils with the sterile mutant of 'Coral Pink' (SC) and its original fertile one (FC). As the results, although SC has functional male and female gametes, few self-pollinated flowers carry out seed-set, leading to sterile phenotype. Additionally, the sterile mechanism of SC was compared with the wild species V. peruviana (VP) which is one of origin of Temari series and exhibits sterility. Interestingly, similar phenotype was observed in PV. We further investigated, therefore, whether there are differences on self-incompatible reaction between SC and VP. (author)
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Energy Technology Data Exchange (ETDEWEB)
Saito, H; Hayashi, Y; Abe, T [RIKEN, Wako (Japan); Kanaya, T; Suzuki, K [Suntory Flowers Ltd., Higashiomi (Japan)
2005-07-01
Full text: Garden verbenas (Verbena hybrida) belonging to the Verbenaceae family are originated from interspecific hybridization among several species, many cultivars frequently produce seeds. Recently, a sterile mutant has been isolated in the verbena cultivar 'Coral Pink' of Temari series (Suntory Flowers Ltd., Tokyo, Japan) by mutation induction using heavy-ion beams at RIKEN Accelarator Research Facility (RARF, Saitama, Japan). We investigated pollen and ovule fertility assessed by acetocarmin staining, seed-set following controlled-pollination tests and behavior of pollen tubes in pistils with the sterile mutant of 'Coral Pink' (SC) and its original fertile one (FC). As the results, although SC has functional male and female gametes, few self-pollinated flowers carry out seed-set, leading to sterile phenotype. Additionally, the sterile mechanism of SC was compared with the wild species V. peruviana (VP) which is one of origin of Temari series and exhibits sterility. Interestingly, similar phenotype was observed in PV. We further investigated, therefore, whether there are differences on self-incompatible reaction between SC and VP. (author)
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Directory of Open Access Journals (Sweden)
Tang Jun
2015-09-01
Full Text Available To determine the characteristics and potential practical applications of glossy cabbage (Brassica oleracea var. capitata L. mutants, five different glossy mutants were studied. The amount of epicuticular wax covering the mutant leaves was only approximately 30% that of the wild-type (WT leaves. The wax crystals of WT plants were columnar and linear, while they were granular and rod-shaped in the mutants. Additionally, in WT cabbage, the primary wax components were alkanes, alcohols, fatty acids, ketones, and aldehydes. There was a significant decrease in the abundance of alkanes and ketones in the wax of the mutants. The glossy-green trait of the mutants may be the result of an inhibited alkane-forming pathway. Higher rates of chlorophyll leaching and water loss demonstrate that the mutant leaves were more permeable and sensitive to drought stress than the WT leaves. Growth curve results indicated that the growth rate of mutant-1 and mutant-3 was slower than that of the corresponding WT cabbage, resulting in shorter plants. However, the growth rate of mutant-2 was not influenced by the lack of coating wax. An investigation of the agronomic traits and heterosis of the glossy cabbage mutants indicated that all five mutants had glossy-green leaves, which was a favorable characteristic. The F1 plants derived from crosses involving mutant-2 exhibited obvious heterosis, suggesting the observed glossy-green trait is controlled by a dominant gene. Therefore, mutant-2 may be useful as a source of genetic material for future cabbage breeding experiments.
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21 CFR 820.184 - Device history record.
2010-04-01
... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Device history record. 820.184 Section 820.184 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES QUALITY SYSTEM REGULATION Records § 820.184 Device history record. Each manufacturer shall...
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46 CFR 184.704 - Marine sanitation devices.
2010-10-01
... 46 Shipping 7 2010-10-01 2010-10-01 false Marine sanitation devices. 184.704 Section 184.704... TONS) VESSEL CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Miscellaneous § 184.704 Marine sanitation devices. A vessel with installed toilet facilities must have a marine sanitation device that complies with...
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Istrate, Alena; Katolik, Adam; Istrate, Andrei; Leumann, Christian J
2017-08-01
We describe the synthesis, thermal stability, structural and RNase H activation properties of 2'β-fluoro-tricyclo nucleic acids (2'F-tc-ANA). Three 2'F-tc-ANA nucleosides (T, 5Me C and A) were synthesized starting from a previously described fluorinated tricyclo sugar intermediate. NMR analysis and quantum mechanical calculations indicate that 2'F-tc-ANA nucleosides prefer sugar conformations in the East and South regions of the pseudorotational cycle. UV-melting experiments revealed that non-consecutive insertions of 2'F-tc-ANA units in DNA reduce the affinity to DNA and RNA complements. However, an oligonucleotide with five contiguous 2'F-tc-ANA-T insertions exhibits increased affinity to complementary RNA. Moreover, a fully modified 10-mer 2'F-tc-ANA oligonucleotide paired to both DNA (+1.6 °C/mod) and RNA (+2.5 °C/mod) with significantly higher affinity compared to corresponding unmodified DNA, and similar affinity compared to corresponding tc-DNA. In addition, CD spectroscopy and molecular dynamics simulations indicate that the conformation of the 2'F-tc-ANA/RNA duplex is similar to that of a DNA/RNA duplex. Moreover, in some sequence contexts, 2'F-tc-ANA promotes RNase H-mediated cleavage of a complementary RNA strand. Taken together, 2'F-tc-ANA represents a nucleic acid analogue that offers the advantage of high RNA affinity while maintaining the ability to activate RNase H, and can be considered a prospective candidate for gene silencing applications. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
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Ueshima, Junichi; Shoji, Mikio; Ratnayake, Dinath B; Abe, Kihachiro; Yoshida, Shinichi; Yamamoto, Kenji; Nakayama, Koji
2003-03-01
The periodontopathogen Porphyromonas gingivalis is an obligate anaerobe that is devoid of catalase but exhibits a relatively high degree of resistance to peroxide stress. In the present study, we demonstrate that P. gingivalis contains a Dps homologue that plays an important role in the protection of cells from peroxide stress. The Dps protein isolated from P. gingivalis displayed a ferritin-like spherical polymer consisting of 19-kDa subunits. Molecular cloning and sequencing of the gene encoding this protein revealed that it had a high similarity in nucleotide and amino acid sequences to Dps proteins from other species. The expression of Dps was significantly increased by exposure of P. gingivalis to atmospheric oxygen in an OxyR-dependent manner, indicating that it is regulated by the reactive oxygen species-regulating gene oxyR. The Dps-deficient mutants, including the dps single mutant and the ftn dps double mutant, showed no viability loss upon exposure to atmospheric oxygen for 6 h. In contrast to the wild type, however, these mutants exhibited the high susceptibility to hydrogen peroxide, thereby disrupting the viability. On the other hand, no significant difference in sensitivity to mitomycin C and metronidazole was observed between the wild type and the mutants. Furthermore, the dps single mutant, compared with the wild type, showed a lower viability in infected human umbilical vein endothelial cells.
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2010-10-01
... within one mile of land must be fitted with a FCC Type Accepted general marine radar system for surface... Federal Communications Commission (FCC) type accepted general marine radar system for surface navigation... 46 Shipping 7 2010-10-01 2010-10-01 false Radars. 184.404 Section 184.404 Shipping COAST GUARD...
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10 CFR 72.184 - Safeguards contingency plan.
2010-01-01
... 10 Energy 2 2010-01-01 2010-01-01 false Safeguards contingency plan. 72.184 Section 72.184 Energy... Protection § 72.184 Safeguards contingency plan. (a) The requirements of the licensee's safeguards contingency plan for responding to threats and radiological sabotage must be as defined in appendix C to part...
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21 CFR 184.1259 - Cocoa butter substitute.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Cocoa butter substitute. 184.1259 Section 184.1259... FOR HUMAN CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 184.1259 Cocoa butter substitute. (a) The common or...
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Both AtrbohD and AtrbohF are essential for mediating responses to oxygen deficiency in Arabidopsis.
Liu, Bo; Sun, Lirong; Ma, Liya; Hao, Fu-Shun
2017-06-01
Both AtrbohD and AtrbohF promote the increases in activities of ADH, PDC, LDH, and Ca 2+ levels, and induce the expression of multiple hypoxia response genes, thus improving Arabidopsis adaptation to oxygen deficiency. NADPH oxidase AtrbohD and AtrbohF cooperatively play key roles in regulation of growth and stress signaling in Arabidopsis. However, reports on AtrbohD and AtrbohF functioning together in hypoxia signaling are scarce, and the underlying mechanisms remain elusive. Here, we show that the double null mutant atrbohD/F is more sensitive to oxygen deprivation compared with wild type (WT) and the single mutant atrbohD and atrbohF. Under oxygen deficiency, enhancements of the transcripts of alcohol dehydrogenase 1 (ADH1) and pyruvate decarboxylase 1 (PDC1) and the activities of ADH, PDC and lactate dehydrogenase in WT are clearly reduced in the single mutants, and more strongly reduced in the double mutant. Moreover, increases in the production of ATP, H 2 O 2 and Ca 2+ in WT are significantly arrested in atrbohD, atrbohF, and especially in atrbohD/F. Hypoxia-promoted rise in the expression of some hypoxic responsive genes is also inhibited in atrbohD/F relative to WT, atrbohD and atrbohF. These genes include ethylene response factor 73, lactate dehydrogenase, MYB transcription factor 2, sucrose synthase 1 (SUS1), SUS4, heat stress transcription factor A2 and heat-shock protein 18.2. These results suggest that both AtrbohD and AtrbohF are essential for mediating hypoxia signaling. H 2 O 2 derived from AtrbohD and AtrbohF triggers the Ca 2+ increase and induces the expression of multiple hypoxia response genes, thus improving Arabidopsis tolerance to low-oxygen stress. These findings provide new insights into the mechanisms of AtrbohF in regulating the responses to oxygen deprivation in Arabidopsis.
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21 CFR 184.1983 - Bakers yeast extract.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Bakers yeast extract. 184.1983 Section 184.1983... Listing of Specific Substances Affirmed as GRAS § 184.1983 Bakers yeast extract. (a) Bakers yeast extract... a selected strain of yeast, Saccharomyces cerevisiae. It may be concentrated or dried. (b) The...
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2010-04-01
...: Category of food Maximum treatment level in food Functional use Bottled water that prior to ozonation meets... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Ozone. 184.1563 Section 184.1563 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN...
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2010-10-01
... 50 Wildlife and Fisheries 7 2010-10-01 2010-10-01 false Mitigation. 216.184 Section 216.184 Wildlife and Fisheries NATIONAL MARINE FISHERIES SERVICE, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION... the coast from 47°07′ N. to 48°30′ N. latitude December January, March and May. (9) Flower Garden...
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SUBMILLIMETER-WAVE ROTATIONAL SPECTROSCOPY OF H2F+
International Nuclear Information System (INIS)
Fujimori, R.; Kawaguchi, K.; Amano, T.
2011-01-01
Five pure rotational transitions of H 2 F + generated by a discharge in an HF/H 2 /Ar mixture were observed in the range 473-774 GHz with a backward-wave oscillator based submillimeter-wave spectrometer. A simultaneous analysis of the rotational lines with 120 combination differences for the ground state derived from the infrared spectra was carried out to determine the precise molecular constants for the ground state. The rotational transition frequencies that lie below 2 THz were calculated, together with their estimated uncertainties, to facilitate future astronomical identifications. The chemistry for H 2 F + formation in interstellar space is discussed in comparison with a case for recently detected H 2 Cl + .
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Present status of two R.F. heating schemes: I.C.R.H. and L.H.R.H
International Nuclear Information System (INIS)
Consoli, T.
1977-01-01
Among the large number of wave-plasma interaction, Ion-Cyclotron Resonant Heating (I.C.R.H.) and Lower Hybrid Resonant Heating (L.H.R.H.), are two promising additional R.F. heating schemes for toroidal hot plasma. They both offer the advantage of using power generators which requires a moderate development for next generation machines. It seems important to try to state in the limits of this paper the present experimental situation of these two R.F. heating methods as it results from the vast literature published from the last European Conference
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46 CFR 184.710 - First-aid kits.
2010-10-01
... 46 Shipping 7 2010-10-01 2010-10-01 false First-aid kits. 184.710 Section 184.710 Shipping COAST... CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Miscellaneous § 184.710 First-aid kits. A vessel must carry either a first-aid kit approved under approval series 160.041 or a kit with equivalent contents...
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21 CFR 184.1034 - Catalase (bovine liver).
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Catalase (bovine liver). 184.1034 Section 184.1034... Listing of Specific Substances Affirmed as GRAS § 184.1034 Catalase (bovine liver). (a) Catalase (bovine liver) (CAS Reg. No. 81457-95-6) is an enzyme preparation obtained from extracts of bovine liver. It is...
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Molecular Genetic Identification Of Some Flax Mutants
International Nuclear Information System (INIS)
AMER, I.M.; MOUSTAFA, H.A.M.
2009-01-01
Five flax genotypes (Linum usitatissimum L.) i.e., commercial cultivar Sakha 2, the mother variety Giza 4 and three mutant types induced by gamma rays, were screened for their salinity tolerance in field experiments (salinity concentration was 8600 and 8300 ppm for soil and irrigation water, respectively). Mutation 6 was the most salt tolerant as compared to the other four genotypes.RAPD technique was used to detect some molecular markers associated with salt tolerance in flax (Mut 6), RAPD-PCR results using 12 random primers exhibited 149 amplified fragments; 91.9% of them were polymorphic and twelve molecular markers (8.1%) for salt tolerant (mutant 6) were identified with molecular size ranged from 191 to 4159 bp and only eight primers successes to amplify these specific markers. Concerning the other mutants, Mut 15 and Mut 25 exhibited 4.3% and 16.2% specific markers, respectively. The induced mutants exhibited genetic similarity to the parent variety were about 51%, 58.3% and 61.1% for Mut 25, Mut 6 and Mut 15, respectively. These specific markers (SM) are used for identification of the induced mutations and it is important for new variety registration.
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Kakita, Veera Mohana Rao; Rachineni, Kavitha; Hosur, Ramakrishna V
2017-07-21
The present manuscript focuses on fast and simultaneous determination of 1 H- 1 H and 1 H- 19 F scalar couplings in fluorinated complex steroid molecules. Incorporation of broadband PSYCHE homonuclear decoupling in the indirect dimension of zero-quantum filtered diagonal experiments (F1-PSYCHE-DIAG) suppresses 1 H- 1 H scalar couplings; however, it retains 1 H- 19 F scalar couplings (along F1 dimension) for the 19 F coupled protons while preserving the pure-shift nature for 1 H resonances uncoupled to 19 F. In such cases, along the direct dimensions, 1 H- 1 H scalar coupling multiplets deconvolute and they appear as duplicated multiplets for the 19 F coupled protons, which facilitates unambiguous discrimination of 19 F coupled 1 H chemical sites from the others. Further, as an added advantage, data acquisition has been accelerated by invoking the known ideas of spectral aliasing in the F1-PSYCHE-DIAG scheme and experiments demand only ~10 min of spectrometer times. Copyright © 2017 John Wiley & Sons, Ltd.
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Hemenway, C S; Dolinski, K; Cardenas, M E; Hiller, M A; Jones, E W; Heitman, J
1995-11-01
We have characterized a Saccharomyces cerevisiae mutant strain that is hypersensitive to cyclosporin A (CsA) and FK506, immunosuppressants that inhibit calcineurin, a serine-threonine-specific phosphatase (PP2B). A single nuclear mutation, designated cev1 for calcineurin essential for viability, is responsible for the CsA-FK506-sensitive phenotype. The peptidyl-prolyl cis-trans isomerases cyclophilin A and FKBP12, respectively, mediate CsA and FK506 toxicity in the cev1 mutant strain. We demonstrate that cev1 is an allele of the VPH6 gene and that vph6 mutant strains fail to assemble the vacuolar H(+)-ATPase (V-ATPase). The VPH6 gene was mapped on chromosome VIII and is predicted to encode a 181-amino acid (21 kD) protein with no identity to other known proteins. We find that calcineurin is essential for viability in many mutant strains with defects in V-ATPase function or vacuolar acidification. In addition, we find that calcineurin modulates extracellular acidification in response to glucose, which we propose occurs via calcineurin regulation of the plasma membrane H(+)-ATPase PMA1. Taken together, our findings suggest calcineurin plays a general role in the regulation of cation transport and homeostasis.
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21 CFR 184.1063 - Enzyme-modified lecithin.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Enzyme-modified lecithin. 184.1063 Section 184.1063... Listing of Specific Substances Affirmed as GRAS § 184.1063 Enzyme-modified lecithin. (a) Enzyme-modified lecithin is prepared by treating lecithin with either phospholipase A2 (EC 3.1.1.4) or pancreatin. (b) The...
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21 CFR 184.1272 - L-Cysteine monohydrochloride.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true L-Cysteine monohydrochloride. 184.1272 Section 184... Listing of Specific Substances Affirmed as GRAS § 184.1272 L-Cysteine monohydrochloride. (a) L-Cysteine... ingredient is used to supply up to 0.009 part of total L-cysteine per 100 parts of flour in dough as a dough...
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Directory of Open Access Journals (Sweden)
Phillip B Grovenstein
2013-07-01
Full Text Available The green micro-alga Chlamydomonas reinhardtii is an elegant model organism to study oxygenic photosynthesis. Chlorophyll (Chl and heme are major tetrapyrroles that play an essential role in photosynthesis and respiration. These tetrapyrroles are synthesized via a common branched pathway that involves mainly enzymes, encoded by nuclear genes. One of the enzymes in the pathway is Mg chelatase (MgChel. MgChel catalyzes insertion of Mg2+ into protoporphyrin IX (PPIX, proto to form Magnesium-protoporphyrin IX (MgPPIX, Mgproto, the first biosynthetic intermediate in the Chl branch. The GUN4 (genomes uncoupled 4 protein is not essential for the MgChel activity but has been shown to significantly stimulate its activity. We have isolated a light sensitive mutant, 6F14, by random DNA insertional mutagenesis. 6F14 cannot tolerate light intensities higher than 90-100 μmol photons m-2 s-1. It shows a light intensity dependent progressive photo-bleaching. 6F14 is incapable of photo-autotrophic growth under light intensity higher than 100 μmol photons m-2 s-1. PCR based analyses show that in 6F14 the insertion of the plasmid outside the GUN4 locus has resulted in a genetic rearrangement of the GUN4 gene and possible deletions in the genomic region flanking the GUN4 gene. Our gun4 mutant has a Chl content very similar to that in the wild type in the dark and is very sensitive to fluctuations in the light intensity in the environment unlike the earlier identified Chlamydomonas gun4 mutant. Complementation with a functional copy of the GUN4 gene restored light tolerance, Chl biosynthesis and photo-autotrophic growth under high light intensities in 6F14. 6F14 is the second gun4 mutant to be identified in C. reinhardtii. Additionally, we show that our two gun4 complements over-express the GUN4 protein and show a higher Chl content per cell compared to that in the wild type strain.
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Teams : M. Brice ; JC Gadmer
2010-01-01
22nd September 2010 - Korean Minister of Education, Science and Technology J.-H. Lee signing the guest book and exchanging gifts with CERN Director-General R. Heuer and Head of International Relations F. Pauss; visiting ALICE exhibition with Collaboration Spokesperson J. Schukraft; accompanied throughout by Adviser R. Voss.
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Kammoonah, Shaima; Prasad, Brinda; Balaraman, Priyadarshini; Mundhada, Hemanshu; Schwaneberg, Ulrich; Plettner, Erika
2018-01-01
Cytochrome P450 cam (a camphor hydroxylase) from the soil bacterium Pseudomonas putida shows potential importance in environmental applications such as the degradation of chlorinated organic pollutants. Seven P450 cam mutants generated from Sequence Saturation Mutagenesis (SeSaM) and isolated by selection on minimal media with either 3-chloroindole or the insecticide endosulfan were studied for their ability to oxidize of 3-chloroindole to isatin. The wild-type enzyme did not accept 3-chloroindole as a substrate. Mutant (E156G/V247F/V253G/F256S) had the highest maximal velocity in the conversion of 3-chloroindole to isatin, whereas mutants (T56A/N116H/D297N) and (G60S/Y75H) had highest k cat /K M values. Six of the mutants had more than one mutation, and within this set, mutation of residues 297 and 179 was observed twice. Docking simulations were performed on models of the mutant enzymes; the wild-type did not accommodate 3-chloroindole in the active site, whereas all the mutants did. We propose two potential reaction pathways for dechlorination of 3-chloroindole. This article is part of a Special Issue entitled: Cytochrome P450 biodiversity and biotechnology, edited by Erika Plettner, Gianfranco Gilardi, Luet Wong, Vlada Urlacher, Jared Goldstone. Copyright © 2017 Elsevier B.V. All rights reserved.
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The h f type particle separators (1961); Les separateurs de particules du type H F (1961)
Energy Technology Data Exchange (ETDEWEB)
Lapostolle, P [Conseil Europeen pour la recherche nucleaire, Div. de la Machine Synchro-Cyclotron, Geneve (Switzerland); Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires
1961-07-01
A general theory is developed for H.F. type separators designed essentially for the analysis of very high energy beams. Analytical expressions are given for the deviation and the separation; these are then presented in the phase space and in the matrix form, thus facilitating the discussion. The particular diagrams proposed by Panofsky, Good and Blewett are described. A few recommendations are made concerning the choice of the device and of the parameters. (author) [French] On developpe une theorie generale des separateurs a H.F., concus essentiellement pour l'analyse de faisceaux de tres haute energie. On presente des expressions analytiques de la deviation et de la separation qui sont ensuite presentees dans l'espace de phase et sous forme matricielle, ce qui en facilite la discussion. On decrit des schemas particuliers proposes par Panofsky, Good et Blewett. Quelques recommandations se degagent pour le choix du dispositif et des parametres. (auteur)
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21 CFR 184.1560 - Ox bile extract.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Ox bile extract. 184.1560 Section 184.1560 Food and... Substances Affirmed as GRAS § 184.1560 Ox bile extract. (a) Ox bile extract (CAS Reg. No. 8008-63-7), also..., partly bitter, disagreeable taste. It is the purified portion of the bile of an ox obtained by...
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Intracellular transport and sorting of mutant human proinsulins that fail to form hexamers.
Quinn, D; Orci, L; Ravazzola, M; Moore, H P
1991-06-01
Human proinsulin and insulin oligomerize to form dimers and hexamers. It has been suggested that the ability of prohormones to self associate and form aggregates may be responsible for the sorting process at the trans-Golgi. To examine whether insulin oligomerization is required for proper sorting into regulated storage granules, we have constructed point mutations in human insulin B chain that have been previously shown to prevent formation of insulin hexamers (Brange, J., U. Ribel, J. F. Hansen, G. Dodson, M. T. Hansen, S. Havelund, S. G. Melberg, F. Norris, K. Norris, L. Snel, A. R. Sorensen, and H. O. Voight. 1988. Nature [Lond.]. 333:679-682). One mutant (B10His----Asp) allows formation of dimers but not hexamers and the other (B9Ser----Asp) prevents formation of both dimers and hexamers. The mutants were transfected into the mouse pituitary AtT-20 cells, and their ability to be sorted into regulated secretory granules was compared to wild-type insulin. We found that while B10His----Asp is sorted somewhat less efficiently than wild-type insulin as reported previously (Carroll, R. J., R. E. Hammer, S. J. Chan, H. H. Swift, A. H. Rubenstein, and D. F. Steiner. 1988. Proc. Natl. Acad. Sci. USA. 85:8943-8947; Gross, D. J., P. A. Halban, C. R. Kahn, G. C. Weir, and L. Villa-Kumaroff. 1989. Proc. Natl. Acad. Sci. USA. 86:4107-4111). B9Ser----Asp is targeted to granules as efficiently as wild-type insulin. These results indicate that self association of proinsulin into hexamers is not required for its targeting to the regulated secretory pathway.
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International Nuclear Information System (INIS)
Huang, Xiaoting; Li, Shu’ni; Zhai, Quanguo; Jiang, Yucheng; Hu, Mancheng
2016-01-01
Graphical abstract: Thermodynamic properties, such as mean activity coefficients, osmotic coefficients and excess Gibbs free energies, of the RbF + RbCl + H 2 O and CsF + CsCl + H 2 O ternary systems were determined from potentiometric measurement at 298.2 K. The Pitzer model and the Harned rule were used to fit the experimental data. - Highlights: • Thermodynamic properties of RbF + RbCl + H 2 O and CsF + CsCl + H 2 O ternary systems were determined. • The Pitzer model and the Harned rule were used to correlate the experimental data. • The mean activity coefficients, osmotic coefficients, and the excess Gibbs free energy were also obtained. - Abstract: Thermodynamic properties of (RbF + RbCl + H 2 O) and (CsF + CsCl + H 2 O) systems were determined by the potentiometric method for different ionic strength fractions y B of RbCl/CsCl at 298.2 K. The Pitzer model and the Harned rule were used to fit the experimental values. The Pitzer mixing parameters and the Harned coefficients were evaluated. In addition, the mean ionic activity coefficients of RbF/CsF and RbCl/CsCl, the osmotic coefficients, and the excess Gibbs energies of the systems studied were calculated.
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Kim, Ha Kun; Chung, Youn Wook; Chock, P Boon; Yim, Moon B
2011-05-15
Missense mutations of SOD1 are linked to familial amyotrophic lateral sclerosis (FALS) through a yet-to-be identified toxic-gain-of-function. One of the proposed mechanisms involves enhanced aggregate formation. However, a recent study showed that dual transgenic mice overexpressing both G93A and CCS copper chaperone (G93A/CCS) exhibit no SOD1-positive aggregates yet show accelerated FALS symptoms with enhanced mitochondrial pathology compared to G93A mice. Using a dicistronic mRNA to simultaneously generate hSOD1 mutants, G93A, A4V and G85R, and hCCS in AAV293 cells, we revealed: (i) CCS is degraded primarily via a macroautophagy pathway. It forms a stable heterodimer with inactive G85R, and via its novel copper chaperone-independent molecular chaperone activity facilitates G85R degradation via a macroautophagy-mediated pathway. For active G93A and A4V, CCS catalyzes their maturation to form active and soluble homodimers. (ii) CCS reduces, under non-oxidative conditions, yet facilitates in the presence of H(2)O(2), mitochondrial translocation of inactive SOD1 mutants. These results, together with previous reports showing FALS SOD1 mutants enhanced free radical-generating activity, provide a mechanistic explanation for the observations with G93A/CCS dual transgenic mice and suggest that free radical generation by FALS SOD1, enhanced by CCS, may, in part, be responsible for the FALS SOD1 mutant-linked aggregation, mitochondrial translocation, and degradation. Published by Elsevier Inc.
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International Nuclear Information System (INIS)
Nettles, W.G.; Beraud, R.; Cole, J.D.; Hamilton, J.H.; Ramayya, A.V.; Spejewski, E.H.; Sastry, K.S.R.
1977-01-01
Levels in 184 Au were investigated by e--γ--t and γ--γ--t coincidence techniques. A level scheme with eight definite and two tentative levels is presented; gamma branching ratios are shown. The state at 156 keV was found to have a lifetime of 36 +- 6 ns; for four of the next five levels, T 1 / 2 was less than 2 ns. 2 figures
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19F-labeling of the adenine H2-site to study large RNAs by NMR spectroscopy
International Nuclear Information System (INIS)
Sochor, F.; Silvers, R.; Müller, D.; Richter, C.; Fürtig, B.; Schwalbe, H.
2016-01-01
In comparison to proteins and protein complexes, the size of RNA amenable to NMR studies is limited despite the development of new isotopic labeling strategies including deuteration and ligation of differentially labeled RNAs. Due to the restricted chemical shift dispersion in only four different nucleotides spectral resolution remains limited in larger RNAs. Labeling RNAs with the NMR-active nucleus 19 F has previously been introduced for small RNAs up to 40 nucleotides (nt). In the presented work, we study the natural occurring RNA aptamer domain of the guanine-sensing riboswitch comprising 73 nucleotides from Bacillus subtilis. The work includes protocols for improved in vitro transcription of 2-fluoroadenosine-5′-triphosphat (2F-ATP) using the mutant P266L of the T7 RNA polymerase. Our NMR analysis shows that the secondary and tertiary structure of the riboswitch is fully maintained and that the specific binding of the cognate ligand hypoxanthine is not impaired by the introduction of the 19 F isotope. The thermal stability of the 19 F-labeled riboswitch is not altered compared to the unmodified sequence, but local base pair stabilities, as measured by hydrogen exchange experiments, are modulated. The characteristic change in the chemical shift of the imino resonances detected in a 1 H, 15 N-HSQC allow the identification of Watson–Crick base paired uridine signals and the 19 F resonances can be used as reporters for tertiary and secondary structure transitions, confirming the potential of 19 F-labeling even for sizeable RNAs in the range of 70 nucleotides
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International Nuclear Information System (INIS)
Canama, Alma O.; Galvez, Hayde F.; Tongson, Eden Jane U.; Quilloy, Reynaldo B.; Hautea, Desiree M.
2010-01-01
Tomato (L.) is one of the most important vegetable crops grown worldwide for the fresh vegetable market and food processing industry. With the completion of the genome-sequencing projects in various crops, the major challenge will be determine the gene function. One approach is to generate and to analyze mutant phenotypes. The paper reports the generation of gamma-irradiated and ethy methane sulfonate (EMS)-treated mutant populations, identification and phenotypic characterization of dominant and visible mutations in tomato mutant lines. Mutant populations of tomato H7996 were created using physical (cobalt 60 gamma ray) and chemical EMS mutagens. Generally, based on high-throughput phenotypic characterization, mutations were observed on the plant habit, size, morphology, leaf and flower color and morphology and fruit characteristics. Specifically, the most common dominant and visible mutations noted in the M 1 generation were monopodial, compact, short internodes, multi-branch plant type, light yellow and ghost leaf coloration, tiny and long pedicel leaf morphology and small or short plant size. In the M2 generation, homogeneous and segregating M 2 families were selected to constitute the core set of visible tomato mutants. Initial bacterial wilt resistance (BWR) gene knockouts were also identified. The mutant lines will be used as a rich source of genetic materials for breeding and functional genomics of tomato. (author)
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The H159A mutant of yeast enolase 1 has significant activity.
Brewer, J M; Holland, M J; Lebioda, L
2000-10-05
The function of His159 in the enolase mechanism is disputed. Recently, Vinarov and Nowak (Biochemistry (1999) 38, 12138-12149) prepared the H159A mutant of yeast enolase 1 and expressed this in Escherichia coli. They reported minimal (ca. 0.01% of the native value) activity, though the protein appeared to be correctly folded, according to its CD spectrum, tryptophan fluorescence, and binding of metal ion and substrate. We prepared H159A enolase using a multicopy plasmid and expressed the enzyme in yeast. Our preparations of H159A enolase have 0.2-0.4% of the native activity under standard assay conditions and are further activated by Mg(2+) concentrations above 1 mM to 1-1.5% of the native activity. Native enolase 1 (and enolase 2) are inhibited by such Mg(2+) concentrations. It is possible that His159 is necessary for correct folding of the enzyme and that expression in E. coli leads to largely misfolded protein. Copyright 2000 Academic Press.
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42 CFR 415.184 - Psychiatric services.
2010-10-01
... 42 Public Health 3 2010-10-01 2010-10-01 false Psychiatric services. 415.184 Section 415.184 Public Health CENTERS FOR MEDICARE & MEDICAID SERVICES, DEPARTMENT OF HEALTH AND HUMAN SERVICES... Psychiatric services. To qualify for physician fee schedule payment for psychiatric services furnished under...
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D'Amour, Pierre; Brossard, Jean-Hugues; Rousseau, Louise; Nguyen-Yamamoto, Loan; Nassif, Edgard; Lazure, Claude; Gauthier, Dany; Lavigne, Jeffrey R; Zahradnik, Richard J
2005-09-01
Non-(1-84) parathyroid hormone (PTH) fragments are large circulating carboxyl-terminal (C) fragments with a partially preserved amino-terminal (N) structure. hPTH (7-84), a synthetic surrogate, has been demonstrated to exert biologic effects in vivo and in vitro which are opposite to those of hPTH (1-34) on the PTH/PTHrP type I receptor through a C-PTH receptor. We wanted to determine the N structure of non-(1-84) PTH fragments. Parathyroid cells isolated from glands obtained at surgery from three patients with primary hyperparathyroidism and three patients with secondary hyperparathyroidism were incubated with 35S-methionine to internally label their secretion products. Incubations were performed for 8 hours at the patient-ionized calcium concentration and in the presence of various protease inhibitors. The supernatant was fractionated by high-performance liquid chromatography (HPLC) and fractions were analyzed with PTH assays having (1 to 4) and (12 to 23) epitopes, respectively. The serum of each patient was similarly analyzed. Peaks of immunoreactivity identified were submitted to sequence analysis to recover the 35S-methionine residues in positions 8 and 18. Three regions of interest were identified with PTH assays. They corresponded to non-(1-84) PTH fragments (further divided in regions 3 and 4), a peak of N-PTH migrating in front of hPTH (1-84) (region 2) and a peak of immunoreactivity corresponding to the elution position of hPTH (1-84) (region 1). The last corresponded to a single sequence starting at position 1. Region 2 gave similar results in all cases (a major signal starting at position 1) but also sometimes minor sequences starting at position 4 or 7. Regions 3 and 4 always identified a major sequence starting at positions 7 and minor sequences starting at positions 8, 10, and 15. Surprisingly, a major signal starting at position 1 was also present in region 3. The HPLC profile obtained from a given patient's parathyroid cells was qualitatively
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Anisimov, Andrey P; Bakhteeva, Irina V; Panfertsev, Evgeniy A; Svetoch, Tat'yana E; Kravchenko, Tat'yana B; Platonov, Mikhail E; Titareva, Galina M; Kombarova, Tat'yana I; Ivanov, Sergey A; Rakin, Alexander V; Amoako, Kingsley K; Dentovskaya, Svetlana V
2009-01-01
Two isogenic sets of Yersinia pestis strains were generated, composed of wild-type strains 231 and I-1996, their non-polar pH 6(-) mutants with deletions in the psaA gene that codes for its structural subunit or the whole operon, as well as strains with restored ability for temperature- and pH-dependent synthesis of adhesion pili or constitutive production of pH 6 antigen. The mutants were generated by site-directed mutagenesis of the psa operon and subsequent complementation in trans. It was shown that the loss of synthesis or constitutive production of pH 6 antigen did not influence Y. pestis virulence or the average survival time of subcutaneously inoculated BALB/c naïve mice or animals immunized with this antigen.
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Directory of Open Access Journals (Sweden)
Benoit-Joseph Laventie
Full Text Available Panton-Valentine leukocidin (PVL, a bicomponent staphylococcal leukotoxin, is involved in the poor prognosis of necrotizing pneumonia. The present study aimed to elucidate the binding mechanism of PVL and in particular its cell-binding domain. The class S component of PVL, LukS-PV, is known to ensure cell targeting and exhibits the highest affinity for the neutrophil membrane (Kd∼10(-10 M compared to the class F component of PVL, LukF-PV (Kd∼10(-9 M. Alanine scanning mutagenesis was used to identify the residues involved in LukS-PV binding to the neutrophil surface. Nineteen single alanine mutations were performed in the rim domain previously described as implicated in cell membrane interactions. Positions were chosen in order to replace polar or exposed charged residues and according to conservation between leukotoxin class S components. Characterization studies enabled to identify a cluster of residues essential for LukS-PV binding, localized on two loops of the rim domain. The mutations R73A, Y184A, T244A, H245A and Y250A led to dramatically reduced binding affinities for both human leukocytes and undifferentiated U937 cells expressing the C5a receptor. The three-dimensional structure of five of the mutants was determined using X-ray crystallography. Structure analysis identified residues Y184 and Y250 as crucial in providing structural flexibility in the receptor-binding domain of LukS-PV.
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Ivanov, Ana; Kameka, Alexander; Pajak, Agnieszka; Bruneau, Luanne; Beyaert, Ronald; Hernández-Sebastià, Cinta; Marsolais, Frédéric
2012-06-01
Asparaginase catalyzes the degradation of L-asparagine to L-aspartic acid and ammonia, and is implicated in the catabolism of transported asparagine in sink tissues of higher plants. The Arabidopsis genome includes two genes, ASPGA1 and ASPGB1, belonging to distinct asparaginase subfamilies. Conditions of severe nitrogen limitation resulted in a slight decrease in seed size in wild-type Arabidopsis. However, this response was not observed in a homozygous T-DNA insertion mutant where ASPG genes had been inactivated. Under nitrogen-sufficient conditions, the ASPG mutant had elevated levels of free asparagine in mature seed. This phenotype was observed exclusively under conditions of low illumination, when a low ratio of carbon to nitrogen was translocated to the seed. Mutants deficient in one or both asparaginases were more sensitive than wild-type to inhibition of primary root elongation and root hair emergence by L-asparagine as a single nitrogen source. This enhanced inhibition was associated with increased accumulation of asparagine in the root of the double aspga1-1/-b1-1 mutant. This indicates that inhibition of root growth is likely elicited by asparagine itself or an asparagine-derived metabolite, other than the products of asparaginase, aspartic acid or ammonia. During germination, a fusion between the ASPGA1 promoter and beta-glucuronidase was expressed in endosperm cells starting at the micropylar end. Expression was initially high throughout the root and hypocotyl, but became restricted to the root tip after three days, which may indicate a transition to nitrogen-heterotrophic growth.
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A novel mouse Fgfr2 mutant, hobbyhorse (hob, exhibits complete XY gonadal sex reversal.
Directory of Open Access Journals (Sweden)
Pam Siggers
Full Text Available The secreted molecule fibroblast growth factor 9 (FGF9 plays a critical role in testis determination in the mouse. In embryonic gonadal somatic cells it is required for maintenance of SOX9 expression, a key determinant of Sertoli cell fate. Conditional gene targeting studies have identified FGFR2 as the main gonadal receptor for FGF9 during sex determination. However, such studies can be complicated by inefficient and variable deletion of floxed alleles, depending on the choice of Cre deleter strain. Here, we report a novel, constitutive allele of Fgfr2, hobbyhorse (hob, which was identified in an ENU-based forward genetic screen for novel testis-determining loci. Fgr2hob is caused by a C to T mutation in the invariant exon 7, resulting in a polypeptide with a mis-sense mutation at position 263 (Pro263Ser in the third extracellular immunoglobulin-like domain of FGFR2. Mutant homozygous embryos show severe limb and lung defects and, when on the sensitised C57BL/6J (B6 genetic background, undergo complete XY gonadal sex reversal associated with failure to maintain expression of Sox9. Genetic crosses employing a null mutant of Fgfr2 suggest that Fgr2hob is a hypomorphic allele, affecting both the FGFR2b and FGFR2c splice isoforms of the receptor. We exploited the consistent phenotype of this constitutive mutant by analysing MAPK signalling at the sex-determining stage of gonad development, but no significant abnormalities in mutant embryos were detected.
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Higuchi, Robert I; Thompson, Anthony W; Chen, Jyun-Hung; Caferro, Thomas R; Cummings, Marquis L; Deckhut, Charlotte P; Adams, Mark E; Tegley, Christopher M; Edwards, James P; López, Francisco J; Kallel, E Adam; Karanewsky, Donald S; Schrader, William T; Marschke, Keith B; Zhi, Lin
2007-10-01
A series of androgen receptor modulators based on 8H-[1,4]oxazino[2,3-f]quinolin-8-ones was synthesized and evaluated in an androgen receptor transcriptional activation assay. The most potent analogues from the series exhibited single-digit nanomolar potency in vitro. Compound 18h demonstrated full efficacy in the maintenance of muscle weight, at 10 mg/kg, with reduced activity in prostate weight in an in vivo model of androgen action.
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Plakortide F acid (PFA) is a marine-derived polyketide endoperoxide exhibiting strong inhibitory activity against several clinically important fungal pathogens. In the present study, transcriptional profiling coupled with mutant and biochemical analyses were conducted using the model organism Sacch...
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Wise, Alexandria; Tenezaca, Luis; Fernandez, Robert W; Schatoff, Emma; Flores, Julian; Ueda, Atsushi; Zhong, Xiaotian; Wu, Chun-Fang; Simon, Anne F; Venkatesh, Tadmiri
2015-01-01
Autism spectrum disorder (ASD) is a neurodevelopmental disorder in humans characterized by complex behavioral deficits, including intellectual disability, impaired social interactions, and hyperactivity. ASD exhibits a strong genetic component with underlying multigene interactions. Candidate gene studies have shown that the neurobeachin (NBEA) gene is disrupted in human patients with idiopathic autism ( Castermans et al., 2003 ). The NBEA gene spans the common fragile site FRA 13A and encodes a signal scaffold protein ( Savelyeva et al., 2006 ). In mice, NBEA has been shown to be involved in the trafficking and function of a specific subset of synaptic vesicles. ( Medrihan et al., 2009 ; Savelyeva et al., 2006 ). Rugose (rg) is the Drosophila homolog of the mammalian and human NBEA. Our previous genetic and molecular analyses have shown that rg encodes an A kinase anchor protein (DAKAP 550), which interacts with components of the epidermal growth factor receptor or EGFR and Notch-mediated signaling pathways, facilitating cross talk between these and other pathways ( Shamloula et al., 2002 ). We now present functional data from studies on the larval neuromuscular junction that reveal abnormal synaptic architecture and physiology. In addition, adult rg loss-of-function mutants exhibit defective social interactions, impaired habituation, aberrant locomotion, and hyperactivity. These results demonstrate that Drosophila NBEA (rg) mutants exhibit phenotypic characteristics reminiscent of human ASD and thus could serve as a genetic model for studying ASDs.
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46 CFR 184.620 - Propulsion engine control systems.
2010-10-01
... 46 Shipping 7 2010-10-01 2010-10-01 false Propulsion engine control systems. 184.620 Section 184... Communications Systems § 184.620 Propulsion engine control systems. (a) A vessel must have two independent means of controlling each propulsion engine. Control must be provided for the engine speed, direction of...
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International Nuclear Information System (INIS)
Buragohain, Amlan; Couck, Sarah; Van Der Voort, Pascal; Denayer, Joeri F.M.; Biswas, Shyam
2016-01-01
Four existing and three new functionalized chromium terephthalates having MIL-101 topology and denoted as Cr-MIL-101-X (existing ones with X=–F, 1-F; –Cl, 2-Cl; –Br, 3-Br; –CH 3 , 4-CH 3 ; new ones with X=–C 6 H 4 , 5-C 6 H 4 ; –F 2 , 6-F 2 , –(CH 3 ) 2 , 7-(CH 3 ) 2 ) were synthesized under hydrothermal conditions. All the materials except 5-C 6 H 4 could be prepared by a general synthetic route, in which the mixtures of CrO 3 , H 2 BDC-X (BDC=1,4-benzenedicarboxylate) linkers, conc. HCl and water with a molar ratio of 1:1:3.9:222.2 were reacted at 180 °C for 144 h. Compared to the 144 h of synthesis time, three of the compounds, namely 1-Cl, 2-Br and 5-C 6 H 4 , could be prepared in much shorter reaction times (12–18 h at 180–210 °C). The materials possess high thermal stability up to 270–300 °C in an air atmosphere. The activated compounds exhibit significant porosity (S BET range: 1273–2135 m 2 g −1 ). At 0 °C and 1 bar, the CO 2 adsorption capacities of the compounds fall in the 1.7–2.9 mmol g −1 range. Compounds 1-F and 6-F 2 showed enhanced CO 2 uptake values compared to parent Cr-MIL-101. The benzene adsorption capacities of the compounds lie in the range of 66.2–139.5 molecules per unit cell at 50 °C and p/p 0 =0.35. The increased benzene uptake value of 1-F compared to un-functionalized Cr-MIL-101 and 4-CH 3 suggests that the fluorination has induced more hydrophobicity in Cr-MIL-101 as compared to the methylation. - Graphical abstract: Benzene adsorption by seven functionalized Cr-MIL-101-X metal-organic framework (MOF) materials Display Omitted - Highlights: • Seven functionalized Cr-MIL-101-X materials were synthesized solvothermally. • All Cr-MIL-101-X materials exhibited high thermal stability up to 270–300 °C in air. • All Cr-MIL-101-X compounds displayed considerable porosity towards N 2 , CO 2 and benzene. • Mono- and di-fluorinated Cr-MIL-101 materials showed enhanced CO 2 adsorption capacities.
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21 CFR 184.1619 - Potassium carbonate.
2010-04-01
... solution of potassium hydroxide with excess carbon dioxide to produce potassium carbonate; (3) By treating a solution of potassium hydroxide with carbon dioxide to produce potassium bicarbonate, which is... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Potassium carbonate. 184.1619 Section 184.1619 Food...
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Directory of Open Access Journals (Sweden)
Lisa A Lewis
2010-07-01
Full Text Available Complement forms an important arm of innate immunity against invasive meningococcal infections. Binding of the alternative complement pathway inhibitor factor H (fH to fH-binding protein (fHbp is one mechanism meningococci employ to limit complement activation on the bacterial surface. fHbp is a leading vaccine candidate against group B Neisseria meningitidis. Novel mechanisms that meningococci employ to bind fH could undermine the efficacy of fHbp-based vaccines. We observed that fHbp deletion mutants of some meningococcal strains showed residual fH binding suggesting the presence of a second receptor for fH. Ligand overlay immunoblotting using membrane fractions from one such strain showed that fH bound to a approximately 17 kD protein, identified by MALDI-TOF analysis as Neisserial surface protein A (NspA, a meningococcal vaccine candidate whose function has not been defined. Deleting nspA, in the background of fHbp deletion mutants, abrogated fH binding and mAbs against NspA blocked fH binding, confirming NspA as a fH binding molecule on intact bacteria. NspA expression levels vary among strains and expression correlated with the level of fH binding; over-expressing NspA enhanced fH binding to bacteria. Progressive truncation of the heptose (Hep I chain of lipooligosaccharide (LOS, or sialylation of lacto-N-neotetraose LOS both increased fH binding to NspA-expressing meningococci, while expression of capsule reduced fH binding to the strains tested. Similar to fHbp, binding of NspA to fH was human-specific and occurred through fH domains 6-7. Consistent with its ability to bind fH, deleting NspA increased C3 deposition and resulted in increased complement-dependent killing. Collectively, these data identify a key complement evasion mechanism with important implications for ongoing efforts to develop meningococcal vaccines that employ fHbp as one of its components.
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Energy Technology Data Exchange (ETDEWEB)
Neveu, G [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires
1961-07-01
We study various compounds resulting from the interaction of UO{sub 2}F{sub 2} with H{sub 2}O and HF (gas), and various triple compounds UO{sub 2}F{sub 2} - H{sub 2}O - HF; the conditions of decomposition and the thermodynamic limits of stability are specified. (author) [French] Nous etudions divers composes formes par reaction de UO{sub 2}F{sub 2} avec H{sub 2}O et HF (gaz) et divers composes triples UO{sub 2}F{sub 2} - H{sub 2}O - HF, en essayant de preciser les decompositions et domaines d'exisfence thermodynamiques de ces corps. (auteur)
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pH-dependent absorption spectra of rhodopsin mutant E113Q: On the role of counterions and protein
Xie, Peng; Zhou, Panwang; Alsaedi, Ahmed; Zhang, Yan
2017-03-01
The absorption spectra of bovine rhodopsin mutant E113Q in solutions were investigated at the molecular level by using a hybrid quantum mechanics/molecular mechanics (QM/MM) method. The calculations suggest the mechanism of the absorption variations of E113Q at different pH values. The results indicate that the polarizations of the counterions in the vicinity of Schiff base under protonation and unprotonation states of the mutant E113Q would be a crucial factor to change the energy gap of the retinal to tune the absorption spectra. Glu-181 residue, which is close to the chromophore, cannot serve as the counterion of the protonated Schiff base of E113Q in dark state. Moreover, the results of the absorption maximum in mutant E113Q with the various anions (Cl-, Br-, I- and NO3-) manifested that the mutant E113Q could have the potential for use as a template of anion biosensors at visible wavelength.
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21 CFR 184.1317 - Garlic and its derivatives.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Garlic and its derivatives. 184.1317 Section 184... Listing of Specific Substances Affirmed as GRAS § 184.1317 Garlic and its derivatives. (a) Garlic is the... derivatives include essential oils, oleo-resins, and natural extractives obtained from garlic. (b) Garlic oil...
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21 CFR 184.1866 - High fructose corn syrup.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true High fructose corn syrup. 184.1866 Section 184.1866... Listing of Specific Substances Affirmed as GRAS § 184.1866 High fructose corn syrup. (a) High fructose... partial enzymatic conversion of glucose (dextrose) to fructose using an insoluble glucose isomerase enzyme...
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Chen, Qi; Cheng, Xiaolin; Wei, Dongqing; Xu, Qin
2015-03-01
Although Elvitegravir (EVG) is a newly developed antiretrovirals drug to treat the acquired immunodeficiency syndrome (AIDS), drug resistance has already been found in clinic, such as E92Q/N155H and Q148H/G140S. Several structural investigations have already been reported to reveal the molecular mechanism of the drug resistance. As full length crystal structure for HIV-1 integrase is still unsolved, we herein use the crystal structure of the full length prototype foamy virus (PFV) in complex with virus DNA and inhibitor Elvitegravir as a template to construct the wild type and E92Q/N155H mutant system of HIV-1 integrase. Molecular dynamic simulations was used to revel the binding mode and the drug resistance of the EVG ligand in E92Q/N155H. Several important interactions were discovered between the mutated residues and the residues in the active site of the E92Q/N155H double mutant pattern, and cross correlation and clustering methods were used for detailed analysis. The results from the MD simulation studies will be used to guide the experimental efforts of developing novel inhibitors against drug-resistant HIV integrase mutants.
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Van Oosten, Michael James
2013-08-08
The accumulation of anthocyanins in plants exposed to salt stress has been largely documented. However, the functional link and regulatory components underlying the biosynthesis of these molecules during exposure to stress are largely unknown. In a screen of second site suppressors of the salt overly sensitive3-1 (sos3-1) mutant, we isolated the anthocyanin-impaired-response-1 (air1) mutant. air1 is unable to accumulate anthocyanins under salt stress, a key phenotype of sos3-1 under high NaCl levels (120 mM). The air1 mutant showed a defect in anthocyanin production in response to salt stress but not to other stresses such as high light, low phosphorous, high temperature or drought stress. This specificity indicated that air1 mutation did not affect anthocyanin biosynthesis but rather its regulation in response to salt stress. Analysis of this mutant revealed a T-DNA insertion at the first exon of an Arabidopsis thaliana gene encoding for a basic region-leucine zipper transcription factor. air1 mutants displayed higher survival rates compared to wild-type in oxidative stress conditions, and presented an altered expression of anthocyanin biosynthetic genes such as F3H, F3′H and LDOX in salt stress conditions. The results presented here indicate that AIR1 is involved in the regulation of various steps of the flavonoid and anthocyanin accumulation pathways and is itself regulated by the salt-stress response signalling machinery. The discovery and characterization of AIR1 opens avenues to dissect the connections between abiotic stress and accumulation of antioxidants in the form of flavonoids and anthocyanins. © 2013 Springer Science+Business Media Dordrecht.
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21 CFR 184.1293 - Ethyl alcohol.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Ethyl alcohol. 184.1293 Section 184.1293 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE Listing of Specific...
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21 CFR 184.1265 - Cuprous iodide.
2010-04-01
... the following specific limitations: Category of food Maximum treatment level in food Functional use... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Cuprous iodide. 184.1265 Section 184.1265 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN...
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Pilot-scale ultrafiltration testing for the F and H area effluent treatment facility
International Nuclear Information System (INIS)
Kessler, J.L.
1984-01-01
An F and H Area Effluent Treatment Facility (F/H ETF) is being designed to treat low activity aqueous effluents which are produced from F and H Area daily operations. The treatment scheme for the F/H ETF will include pretreatment (pH adjustment and filtration) followed by Reverse Osmosis and/or Ion Exchange to remove dissolved species. Several alternative treatment processes are being considered for the F/H ETF. One of the alternatives in the pretreatment step is tubular Ultrafiltration (UF), using a dynamically formed zirconium oxide membrane supported on a porous stainless steel backing. Pilot-scale testing with a single membrane module (13 ft 2 area) and 200-Area effluent simulant has demonstrated that UF is a viable filtration option for the F/H ETF. UF testing at TNX has defined the operating conditions necessary for extended operation and also demonstrated excellent filtration performance (filtrate SDI 2 /day) flux and will provide excellent pretreatment for both reverse osmosis and ion exchange. 2 refs
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Induction of Mutants in Durum Wheat
International Nuclear Information System (INIS)
AL-Ubaidi, M.; Ibrahim, I.; AL-Hadithi, A.
2002-01-01
This investigation presents a breeding program for induction and development of a new genotype of durum wheat, resistant to lodging with high yield, by irradiation durum wheat hybrids (F2) with gamma rays 100 Gy, during 1990-1997 cultivation seasons. This program involves: induction of variability, selection evaluation of the mutants at three locations: Twaitha (Baghdad) Latifya ( Babylon) and Swari (Kutt). All mutants showed resistance to lodging and there was a significant reduction in plant height. Mutant SIXIZ-22 surpassed other mutants and its origin in lodging resistance and plant height (83.5,82.8 and 89.4 cm) in the three locations at generation M5 and M6, respectively. Also, there were significant differences between mutant and their origin in the number of spikes/M 2 and grain yild during the two successive generation. On the other hand, mutant IZxCO-105 surpassed other mutants in the number of spikes/M 2 (231.8,242.3 and 292) and grain yield (4336,3376 and 5232 kg/ha) in all testing location, respectively . (authors) 14 refs., 4 tabs
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International Nuclear Information System (INIS)
Ryan, J.P.
1984-01-01
The F and H Area Effluent Treatment Facility is essentially a four-stage process that will decontaminate the waste water that is currently being discharged to seepage basins in the Separations Areas. The stages include pretreatment, reverse osmosis, ion exchange, and evaporation. A series of tests were performed at Carre, Inc. (Seneca, SC) from March 5 through March 13, to determine the usefulness of ultrafiltration (UF) in the pretreatment stage of the ETF. The results of that testing program indicate that UF would be an excellent means of removing entrained activity from the 200 Area process effluents. Hyperfiltration (HF) was also tested as a means of providing an improved concentration factor from the reverse osmosis stage. The results show that the membranes that were tested would not reject salt well enough at high salt concentrations to be useful in the final reverse osmosis stage. However, there are several membranes which are commercially available that would provide the needed rejection if they could be applied (dynamically) on the Carre support structure. This avenue is still being explored, as theoretically, it could eliminate the need for the F/H ETF evaporator
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21 CFR 184.1979a - Reduced lactose whey.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Reduced lactose whey. 184.1979a Section 184.1979a... Listing of Specific Substances Affirmed as GRAS § 184.1979a Reduced lactose whey. (a) Reduced lactose whey is the substance obtained by the removal of lactose from whey. The lactose content of the finished...
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21 CFR 184.1979b - Reduced minerals whey.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Reduced minerals whey. 184.1979b Section 184.1979b... Listing of Specific Substances Affirmed as GRAS § 184.1979b Reduced minerals whey. (a) Reduced minerals whey is the substance obtained by the removal of a portion of the minerals from whey. The dry product...
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21 CFR 184.1187 - Calcium alginate.
2010-04-01
... ingredient is used in food only within the following specific limitations: Category of food Maximum level of... other food categories 0.3 Do. (d) Prior sanctions for calcium alginate different from the uses... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Calcium alginate. 184.1187 Section 184.1187 Food...
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21 CFR 184.1120 - Brown algae.
2010-04-01
... used in food only within the following specific limitations: Category of food Maximum level of use in... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Brown algae. 184.1120 Section 184.1120 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN...
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2010-04-01
... within the following specific limitations: Category of food Maximum level of use in food (as served... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Red algae. 184.1121 Section 184.1121 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN...
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21 CFR 184.1011 - Alginic acid.
2010-04-01
... used in food only within the following specific limitations: Category of food Maximum level of use in... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Alginic acid. 184.1011 Section 184.1011 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN...
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21 CFR 184.1801 - Sodium tartrate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sodium tartrate. 184.1801 Section 184.1801 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... crystals. It is obtained as a byproduct of wine manufacture. (b) The ingredient meets the specifications of...
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Defective Glycinergic Synaptic Transmission in Zebrafish Motility Mutants
Hirata, Hiromi; Carta, Eloisa; Yamanaka, Iori; Harvey, Robert J.; Kuwada, John Y.
2010-01-01
Glycine is a major inhibitory neurotransmitter in the spinal cord and brainstem. Recently, in vivo analysis of glycinergic synaptic transmission has been pursued in zebrafish using molecular genetics. An ENU mutagenesis screen identified two behavioral mutants that are defective in glycinergic synaptic transmission. Zebrafish bandoneon (beo) mutants have a defect in glrbb, one of the duplicated glycine receptor (GlyR) β subunit genes. These mutants exhibit a loss of glycinergic synaptic ...
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{sup 19}F-labeling of the adenine H2-site to study large RNAs by NMR spectroscopy
Energy Technology Data Exchange (ETDEWEB)
Sochor, F. [Johann Wolfgang Goethe-University Frankfurt, Institut für Organische Chemie und Chemische Biologie, Center for Biomolecular Magnetic Resonance (BMRZ) (Germany); Silvers, R. [Massachusetts Institute of Technology, Department of Chemistry, Francis Bitter Magnet Laboratory (United States); Müller, D.; Richter, C.; Fürtig, B., E-mail: fuertig@nmr.uni-frankfurt.de; Schwalbe, H., E-mail: schwalbe@nmr.uni-frankfurt.de [Johann Wolfgang Goethe-University Frankfurt, Institut für Organische Chemie und Chemische Biologie, Center for Biomolecular Magnetic Resonance (BMRZ) (Germany)
2016-01-15
In comparison to proteins and protein complexes, the size of RNA amenable to NMR studies is limited despite the development of new isotopic labeling strategies including deuteration and ligation of differentially labeled RNAs. Due to the restricted chemical shift dispersion in only four different nucleotides spectral resolution remains limited in larger RNAs. Labeling RNAs with the NMR-active nucleus {sup 19}F has previously been introduced for small RNAs up to 40 nucleotides (nt). In the presented work, we study the natural occurring RNA aptamer domain of the guanine-sensing riboswitch comprising 73 nucleotides from Bacillus subtilis. The work includes protocols for improved in vitro transcription of 2-fluoroadenosine-5′-triphosphat (2F-ATP) using the mutant P266L of the T7 RNA polymerase. Our NMR analysis shows that the secondary and tertiary structure of the riboswitch is fully maintained and that the specific binding of the cognate ligand hypoxanthine is not impaired by the introduction of the {sup 19}F isotope. The thermal stability of the {sup 19}F-labeled riboswitch is not altered compared to the unmodified sequence, but local base pair stabilities, as measured by hydrogen exchange experiments, are modulated. The characteristic change in the chemical shift of the imino resonances detected in a {sup 1}H,{sup 15}N-HSQC allow the identification of Watson–Crick base paired uridine signals and the {sup 19}F resonances can be used as reporters for tertiary and secondary structure transitions, confirming the potential of {sup 19}F-labeling even for sizeable RNAs in the range of 70 nucleotides.
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Effects of initial microstructure and helium production on radiation hardening in F82H Steels
Energy Technology Data Exchange (ETDEWEB)
Okubo, N.; Wakai, E.; Takada, F.; Jitsukawa, S. [Japan Atomic Energy Agency, Naga-gun, Ibaraki-ken (Japan); Katoh, Y. [Oak Ridge Noational Laboratory, TN (United States)
2007-07-01
Full text of publication follows: Fission neutron irradiation to steels doped with isotope boron-10 is frequently conducted to study effects of the helium production on mechanical properties. The intrinsic mechanical properties of F82H steels could have been changed due to the boron doping. Recently, we reported that co-doping with boron and nitrogen to F82H (F82H+B+N) improved the mechanical properties of F82H doped only with boron. The mechanical properties of F82H+B+N are successfully comparable with the non-doped F82H before irradiation. In order to evaluate the effects of initial microstructure and helium production on radiation hardening, F82H and F82H+B+N were irradiate d Specimens used in this study were standard F82H martensitic steels, F82H steels doped with 60 mass ppm {sup 10}B and 200 ppm N (F82H+10B+N) and F82H steels doped with 60 mass ppm {sup 11}B and 200 ppm N (F82H+11B+N). Initial microstructures were changed by tempering conditions, and the tempering temperatures were at 700, 750 and 780 deg. C. Irradiation was performed at nominally 250 deg. C to 2 dpa in JMTR. Tensile properties were measured for the specimens before and after irradiation. Change of yield stress due to the irradiation in the F82H+11B+N steels depended strongly on the initial microstructure and hardness before irradiation. The radiation hardening due to helium production in the F82H+10B+N steels was less than 60 MPa in these experiments. Size of dimple in the fracture surface of specimen with helium production was larger than that with non-helium production. (authors)
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Directory of Open Access Journals (Sweden)
Xiao-hui ZHEN
2014-11-01
Full Text Available Physiological, biochemical and electron microscopy analyses were used to investigate the photosynthetic performance of flag leaves in rice white stripe mutant 6001 during the senescence process. Results showed that the chlorophyll content at the heading and milk-ripe stages in rice mutant 6001 were about 34.78% and 3.00% less than those in wild type 6028, respectively. However, the chlorophyll content at the fully-ripe stage in rice mutant 6001 was higher than that in wild type 6028. At the heading stage, the net photosynthetic rate (Pn in rice mutant 6001 was lower than that in wild type 6028. Rice mutant 6001 also exhibited a significantly slower decrease rate of Pn than wild type 6028 during the senescence progress, especially at the later stage. Furthermore, Ca2+-ATPase, Mg2+-ATPase and photophosphorylation activities exhibited the similar trends as the Pn. During the senescence process, the 68 kDa polypeptide concentrations in the thylakoid membrane proteins exhibited a significant change, which was one of the critical factors that contributed to the observed change in photosynthesis. We also observed that the chloroplasts of rice mutant 6001 exhibited higher integrity than those of wild type 6028, and the chloroplast membrane of rice mutant 6001 disintegrated more slow during the senescence process. In general, rice mutant 6001 had a relatively slower senescence rate than wild type 6028, and exhibited anti-senescence properties.
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Directory of Open Access Journals (Sweden)
Haiyan Mao
Full Text Available Treatment of LQT2 is inadequate. Many drugs which can pharmacologically rescue defective protein trafficking in LQT2 also result in potent blockade of HERG current, negating their therapeutic benefit. It is reported that PD-118057 and thapsigargin can rescue LQT2 without hERG channel blockade, but the precise mechanism of action is unknown. Furthermore, the effect of PD-118057 and thapsigargin on the dominant negative E637K-hERG mutant has not been previously investigated.IN THIS STUDY, WE INVESTIGATED: (a the effect of PD-118057 and thapsigargin on the current amplitudes of WT-hERG and WT/E637K-hERG channels; (b the effect of PD-118057 and thapsigargin on the biophysical properties of WT-hERG and WT/E637K-hERG channels; (c whether drug treatment can rescue channel processing and trafficking defects of the WT/E637K-hERG mutant.The whole-cell Patch-clamp technique was used to assess the effect of PD-118057 and thapsigargin on the electrophysiological characteristics of the rapidly activating delayed rectifier K(+ current (Ikr of the hERG protein channel. Western blot was done to investigate pharmacological rescue on hERG protein channel function.In our study, PD-118057 was shown to significantly enhance both the maximum current amplitude and tail current amplitude, but did not alter the gating and kinetic properties of the WT-hERG channel, with the exception of accelerating steady-state inactivation. Additionally, thapsigargin shows a similar result as PD-118057 for the WT-hERG channel, but with the exception of attenuating steady-state inactivation. However, for the WT/E637K-hERG channel, PD-118057 had no effect on either the current or on the gating and kinetic properties. Furthermore, thapsigargin treatment did not alter the current or the gating and kinetic properties of the WT/E637K-hERG channel, with the exception of opening at more positive voltages.Our findings illustrate that neither PD-118057 nor thapsigargin play a role in correcting
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F/H Effluent Treatment Facility filtration upgrade alternative evaluations overview
Energy Technology Data Exchange (ETDEWEB)
Miles, W.C. Jr.; Poirier, M.R.; Brown, D.F.
1992-01-01
The F/H Effluent Treatment Facility (ETF) at the Savannah River Site (SRS) was designed to treat process wastewater from the 200-F/H Production Facilities (routine wastewater) as well as intermittent flows from the F/H Retention Basins and F/H Cooling Water Basins (nonroutine wastewater). Since start-up of the ETF at SRS in 1988, the treatment process has experienced difficulties processing routine and nonroutine wastewater. Studies have identified high bacteria and bacterial decomposition products in the wastewater as the cause for excessive fouling of the filtration system. In order to meet Waste Management requirements for the treatment of processed wastewater, an upgrade of the ETF filtration system is being developed. This upgrade must be able to process the nonroutine wastewater at design capacity. As a result, a study of alternative filter technologies was conducted utilizing simulated wastewater. The simulated wastewater tests have been completed. Three filter technologies, centrifugal polymeric ultrafilters, tubular polymeric ultrafilters, and backwashable cartridge filters have been selected for further evaluation utilizing actual ETF wastewater.
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F/H Effluent Treatment Facility filtration upgrade alternative evaluations overview
Energy Technology Data Exchange (ETDEWEB)
Miles, W.C. Jr.; Poirier, M.R.; Brown, D.F.
1992-07-01
The F/H Effluent Treatment Facility (ETF) at the Savannah River Site (SRS) was designed to treat process wastewater from the 200-F/H Production Facilities (routine wastewater) as well as intermittent flows from the F/H Retention Basins and F/H Cooling Water Basins (nonroutine wastewater). Since start-up of the ETF at SRS in 1988, the treatment process has experienced difficulties processing routine and nonroutine wastewater. Studies have identified high bacteria and bacterial decomposition products in the wastewater as the cause for excessive fouling of the filtration system. In order to meet Waste Management requirements for the treatment of processed wastewater, an upgrade of the ETF filtration system is being developed. This upgrade must be able to process the nonroutine wastewater at design capacity. As a result, a study of alternative filter technologies was conducted utilizing simulated wastewater. The simulated wastewater tests have been completed. Three filter technologies, centrifugal polymeric ultrafilters, tubular polymeric ultrafilters, and backwashable cartridge filters have been selected for further evaluation utilizing actual ETF wastewater.
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F/H effluent treatment facility filtration upgrade alternative evaluations overview
International Nuclear Information System (INIS)
Miles, W.C. Jr.; Poirier, M.R.; Brown, D.F.
1992-01-01
The F/H Effluent Treatment Facility (ETF) at the Savannah River Site (SRS) was designed to treat process wastewater from the 200-F/H Production Facilities (routine wastewater) as well as intermittent flows from the F/H Retention Basins and F/H Cooling Water Basins (nonroutine wastewater). Since start-up of the ETF at SRS in 1988, the treatment process has experienced difficulties processing routine and nonroutine wastewater. Studies have identified high bacteria and bacterial decomposition products in the wastewater as the cause for excessive fouling of the filtration system. In order to meet Waste Management requirements for the treatment of processed wastewater, an upgrade of the ETF filtration system is being developed. This upgrade must be able to process the nonroutine wastewater at design capacity. As a result, a study of alternative filter technologies was conducted utilizing simulated wastewater. The simulated wastewater tests have been completed. Three filter technologies, centrifugal polymeric ultrafilters, tubular polymeric ultrafilters, and backwashable cartridge filters have been selected for further evaluation utilizing actual ETF wastewater. (author)
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Energy Technology Data Exchange (ETDEWEB)
Shin, Dae-Seop; Park, Myoung Joo [Drug Discovery Platform Technology Research Group, Korea Research Institute of Chemical Technology, Daejeon (Korea, Republic of); Lee, Hyang-Ae [Korea Institute of Toxicology, Korea Research Institute of Chemical Technology, Daejeon (Korea, Republic of); Lee, Joo Yun; Chung, Hee-Chung; Yoo, Dae Seok; Chae, Chong Hak [Drug Discovery Platform Technology Research Group, Korea Research Institute of Chemical Technology, Daejeon (Korea, Republic of); Park, Sang-Joon [College of Veterinary Medicine, Kyungpook National University, Daegu (Korea, Republic of); Kim, Ki-Suk [Korea Institute of Toxicology, Korea Research Institute of Chemical Technology, Daejeon (Korea, Republic of); Bae, Myung Ae, E-mail: mbae@krict.re.kr [Drug Discovery Platform Technology Research Group, Korea Research Institute of Chemical Technology, Daejeon (Korea, Republic of)
2014-02-01
Nefazodone was used widely as an antidepressant until it was withdrawn from the U.S. market in 2004 due to hepatotoxicity. We have investigated methods to predict various toxic effects of drug candidates to reduce the failure rate of drug discovery. An electrophysiological method was used to assess the cardiotoxicity of drug candidates. Small molecules, including withdrawn drugs, were evaluated using a patch-clamp method to establish a database of hERG inhibition. Nefazodone inhibited hERG channel activity in our system. However, nefazodone-induced hERG inhibition indicated only a theoretical risk of cardiotoxicity. Nefazodone inhibited the hERG channel in a concentration-dependent manner with an IC{sub 50} of 45.3 nM in HEK-293 cells. Nefazodone accelerated both the recovery from inactivation and its onset. Nefazodone also accelerated steady-state inactivation, although it did not modify the voltage-dependent character. Alanine mutants of hERG S6 and pore region residues were used to identify the nefazodone-binding site on hERG. The hERG S6 point mutants Y652A and F656A largely abolished the inhibition by nefazodone. The pore region mutant S624A mildly reduced the inhibition by nefazodone but T623A had little effect. A docking study showed that the aromatic rings of nefazodone interact with Y652 and F656 via π–π interactions, while an amine interacted with the S624 residue in the pore region. In conclusion, Y652 and F656 in the S6 domain play critical roles in nefazodone binding. - Highlights: • Nefazodone inhibits hERG channels with an IC{sub 50} of 45.3 nM in HEK-293 cells. • Nefazodone blocks hERG channels by binding to the open channels. • Y652 and F656 are important for binding of nefazodone. • The aromatic rings of nefazodone interact with Y652 and F656 via π–π interactions.
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21 CFR 184.1613 - Potassium bicarbonate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Potassium bicarbonate. 184.1613 Section 184.1613 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS §...
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21 CFR 184.1610 - Potassium alginate.
2010-04-01
...: Category of food Maximum level of use in food (as served) (percent) Functional use Confections and...) of this chapter 0.25 Do. All other food categories 0.01 Do. (d) Prior sanctions for potassium... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Potassium alginate. 184.1610 Section 184.1610 Food...
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21 CFR 184.1736 - Sodium bicarbonate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sodium bicarbonate. 184.1736 Section 184.1736 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS §...
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21 CFR 184.1099 - Tartaric acid.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Tartaric acid. 184.1099 Section 184.1099 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN..., crystalline powder. It is odorless and has an acid taste. It is obtained as a byproduct of wine manufacture...
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Inhibition of Early Stages of HIV-1 Assembly by INI1/hSNF5 Transdominant Negative Mutant S6 ▿
Cano, Jennifer; Kalpana, Ganjam V.
2011-01-01
INI1/hSNF5 is an HIV-1 integrase (IN) binding protein specifically incorporated into virions. A truncated mutant of INI1 (S6, amino acids 183 to 294) harboring the minimal IN binding Rpt1 domain potently inhibits HIV-1 particle production in a transdominant manner. The inhibition requires interaction of S6 with IN within Gag-Pol. While INI1 is a nuclear protein and harbors a masked nuclear export signal (NES), the transdominant negative mutant S6 is cytoplasmic, due to the unmasking of NES. Here, we examined the effects of subcellular localization of S6 on HIV-1 inhibition and further investigated the stages of assembly that are affected. We found that targeting a nuclear localization signal-containing S6 variant [NLS-S6(Rpt1)] to the nucleoplasm (but not to the nucleolus) resulted in complete reversal of inhibition of particle production. Electron microscopy indicated that although no electron-dense particles at any stage of assembly were seen in cells expressing S6, virions were produced in cells expressing the rescue mutant NLS-S6(Rpt1) to wild-type levels. Immunofluorescence analysis revealed that p24 exhibited a diffuse pattern of localization within the cytoplasm in cells expressing S6 in contrast to accumulation along the membrane in controls. Pulse-chase analysis indicated that in S6-expressing cells, although Gag(Pr55gag) protein translation was unaffected, processing and release of p24 were defective. Together, these results indicate that expression of S6 in the cytoplasm interferes with trafficking of Gag-Pol/Gag to the membrane and causes a defective processing leading to inhibition of assembly at an early stage prior to particle formation and budding. PMID:21159874
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Kovarik, Zrinka; Hrvat, Nikolina Maček; Katalinić, Maja; Sit, Rakesh K.; Paradyse, Alexander; Žunec, Suzana; Musilek, Kamil; Fokin, Valery V.; Taylor, Palmer; Radić, Zoran
2016-01-01
Exposure to the nerve agent soman is difficult to treat due to the rapid dealkylation of soman-acetylcholinesterase (AChE) conjugate known as aging. Oxime antidotes commonly used to reactivate organophosphate inhibited AChE are ineffective against soman, while the efficacy of the recommended nerve agent bioscavenger butyrylcholinesterase is limited by strictly stoichiometric scavenging. To overcome this limitation, we tested ex vivo, in human blood, and in vivo, in soman exposed mice, the capacity of aging-resistant human AChE mutant Y337A/F338A in combination with oxime HI-6 to act as a catalytic bioscavenger of soman. HI-6 was previously shown in vitro to efficiently reactivate this mutant upon soman, as well as VX, cyclosarin, sarin and paraoxon inhibition. We here demonstrate that ex vivo, in whole human blood, 1 μM soman was detoxified within 30 minutes when supplemented with 0.5 μM Y337A/F338A AChE and 100 μM HI-6. This combination was further tested in vivo. Catalytic scavenging of soman in mice improved the therapeutic outcome and resulted in the delayed onset of toxicity symptoms. Furthermore, in a preliminary in vitro screen we identified an even more efficacious oxime than HI-6, in a series of forty-two pyridinium aldoximes, and five imidazole 2-aldoxime N-propyl pyridinium derivatives. One of the later imidazole aldoximes, RS-170B, was a 2–3 –fold more effective reactivator of Y337A/F338A AChE than HI-6 due to the smaller imidazole ring, as indicated by computational molecular models, that affords a more productive angle of nucleophilic attack. PMID:25835984
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Isolation of New Gravitropic Mutants under Hypergravity Conditions.
Mori, Akiko; Toyota, Masatsugu; Shimada, Masayoshi; Mekata, Mika; Kurata, Tetsuya; Tasaka, Masao; Morita, Miyo T
2016-01-01
Forward genetics is a powerful approach used to link genotypes and phenotypes, and mutant screening/analysis has provided deep insights into many aspects of plant physiology. Gravitropism is a tropistic response in plants, in which hypocotyls and stems sense the direction of gravity and grow upward. Previous studies of gravitropic mutants have suggested that shoot endodermal cells in Arabidopsis stems and hypocotyls are capable of sensing gravity (i.e., statocytes). In the present study, we report a new screening system using hypergravity conditions to isolate enhancers of gravitropism mutants, and we also describe a rapid and efficient genome mapping method, using next-generation sequencing (NGS) and single nucleotide polymorphism (SNP)-based markers. Using the endodermal-amyloplast less 1 ( eal1 ) mutant, which exhibits defective development of endodermal cells and gravitropism, we found that hypergravity (10 g) restored the reduced gravity responsiveness in eal1 hypocotyls and could, therefore, be used to obtain mutants with further reduction in gravitropism in the eal1 background. Using the new screening system, we successfully isolated six ene ( enhancer of eal1 ) mutants that exhibited little or no gravitropism under hypergravity conditions, and using NGS and map-based cloning with SNP markers, we narrowed down the potential causative genes, which revealed a new genetic network for shoot gravitropism in Arabidopsis .
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21 CFR 184.1135 - Ammonium bicarbonate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Ammonium bicarbonate. 184.1135 Section 184.1135 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 18...
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21 CFR 184.1660 - Propyl gallate.
2010-04-01
... alcohol. (b) The ingredient meets the specifications of the “Food Chemicals Codex,” 3d Ed. (1981), pp. 257... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Propyl gallate. 184.1660 Section 184.1660 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN...
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21 CFR 184.1133 - Ammonium alginate.
2010-04-01
...: Category of food Maximum level of use in food (as served) (percent) Functional use Confections, frostings... chapter 0.4 Do. Sweet sauces, § 170.3(n)(43) of this chapter 0.5 Do. All other food categories 0.1... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Ammonium alginate. 184.1133 Section 184.1133 Food...
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Fracture toughness of the F-82H steel-effect of loading modes, hydrogen, and temperature
International Nuclear Information System (INIS)
Li, H.-X.; Jones, R.H.; Hirth, J.P.; Gelles, D.S.
1996-01-01
The effects of loading mode, hydrogen, and temperature on fracture toughness and tearing modulus were examined for a ferritic/martensitic steel (F-82H). The introduction of a shear load component, mode III, significantly decreased the initiation and propagation resistance of cracks compared to the opening load, mode I, behavior. Mode I crack initiation and propagation exhibited the highest resistance. A minimum resistance occurred when the mode I and mode III loads were nearly equal. The presence of 4 wppm hydrogen decreased the cracking resistance compared to behavior without H regardless of the loading mode. The minimum mixed-mode fracture toughness with the presence of hydrogen was about 30% of the hydrogen-free mode I fracture toughness. The mixed-mode toughness exhibited a lesser sensitivity to temperature than the mode I toughness. The J IC value was 284 kJ/m 2 at room temperature, but only 60 kJ/m 2 at -55 C and 30 kJ/m 2 at -90 C. The ductile to brittle transition temperature (DBTT) was apparently higher than -55 C. (orig.)
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CtBP1/BARS Gly172 → Glu mutant structure: Impairing NAD(H)-binding and dimerization
International Nuclear Information System (INIS)
Nardini, Marco; Valente, Carmen; Ricagno, Stefano; Luini, Alberto; Corda, Daniela; Bolognesi, Martino
2009-01-01
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.
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Energy Technology Data Exchange (ETDEWEB)
Chen, Qi [Shanghai Jiao Tong Univ., Shanghai (China). State Key Lab. of Microbial Metabolism and College of Life Science and Biotechnology; Cheng, Xiaolin [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Center for Molecular Biophysics; Univ. of Tennessee, Knoxville, TN (United States). Dept. of Biochemistry and Cellular and Molecular Biology; Wei, Dongqing [Shanghai Jiao Tong Univ., Shanghai (China). State Key Lab. of Microbial Metabolism and College of Life Science and Biotechnology; Xu, Qin [Shanghai Jiao Tong Univ., Shanghai (China). State Key Lab. of Microbial Metabolism and College of Life Science and Biotechnology
2014-11-06
Although Elvitegravir (EVG) is a newly developed antiretrovirals drug to treat the acquired immunodeficiency syndrome (AIDS), drug resistance has already been found in clinic, such as E92Q/N155H and Q148H/G140S. Several structural investigations have already been reported to reveal the molecular mechanism of the drug resistance. As full length crystal structure for HIV-1 integrase is still unsolved, we use in this paper the crystal structure of the full length prototype foamy virus (PFV) in complex with virus DNA and inhibitor Elvitegravir as a template to construct the wild type and E92Q/N155H mutant system of HIV-1 integrase. Molecular dynamic simulations was used to revel the binding mode and the drug resistance of the EVG ligand in E92Q/N155H. Several important interactions were discovered between the mutated residues and the residues in the active site of the E92Q/N155H double mutant pattern, and cross correlation and clustering methods were used for detailed analysis. The results from the MD simulation studies will be used to guide the experimental efforts of developing novel inhibitors against drug-resistant HIV integrase mutants.
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Vijayan, Saptha; Mallick, Sathi; Dutta, Mouparna; Narayani, M; Ghosh, Anindya S
2014-02-01
MreB is a cytoskeletal protein, which is responsible for maintaining proper cellular morphology and is essential for cell survival. Likewise, penicillin-binding protein 5 (PBP5) helps in maintaining cell shape, though non-essential for survival. The contradicting feature of these two proteins paves the way for this study, wherein we attempt to draw a relation on the nature of distribution of MreB in PBP deletion mutants. The study revealed that the uniform MreB helices/patches were destabilized/disturbed at the zone of deformities of the PBP mutants, whereas the helical patterns were retained at the regions maintaining a rod shape. We interpret that MreB remains functional irrespective of its distribution being misguided by the aberrant shapes of PBP mutants.
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Shima, Jun; Hino, Akihiro; Yamada-Iyo, Chie; Suzuki, Yasuo; Nakajima, Ryouichi; Watanabe, Hajime; Mori, Katsumi; Takano, Hiroyuki
1999-01-01
Accumulation of trehalose is widely believed to be a critical determinant in improving the stress tolerance of the yeast Saccharomyces cerevisiae, which is commonly used in commercial bread dough. To retain the accumulation of trehalose in yeast cells, we constructed, for the first time, diploid homozygous neutral trehalase mutants (Δnth1), acid trehalase mutants (Δath1), and double mutants (Δnth1 ath1) by using commercial baker’s yeast strains as the parent strains and the gene disruption method. During fermentation in a liquid fermentation medium, degradation of intracellular trehalose was inhibited with all of the trehalase mutants. The gassing power of frozen doughs made with these mutants was greater than the gassing power of doughs made with the parent strains. The Δnth1 and Δath1 strains also exhibited higher levels of tolerance of dry conditions than the parent strains exhibited; however, the Δnth1 ath1 strain exhibited lower tolerance of dry conditions than the parent strain exhibited. The improved freeze tolerance exhibited by all of the trehalase mutants may make these strains useful in frozen dough. PMID:10388673
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21 CFR 184.1724 - Sodium alginate.
2010-04-01
....1(b)(2), the ingredient is used in food only within the following specific limitations: Category of...; texturizer, § 170.3(o)(32) of this chapter. All other food categories 1.0 Emulsifier, § 170.3(o)(8) of this... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sodium alginate. 184.1724 Section 184.1724 Food and...
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Cody, Vivian; Pace, Jim; Piraino, Jennifer; Queener, Sherry F.
2011-01-01
In order to produce a more potent replacement for trimethoprim (TMP) used as a therapy for Pneumocystis pneumonia and targets dihydrofolate reductase from Pneumocystis jirovecii (pjDHFR), it is necessary to understand the determinants of potency and selectivity against DHFR from the mammalian host and fungal pathogen cells. To this end, active site residues in human (h)DHFR were replaced with those from pjDHFR. Structural data are reported for two complexes of TMP with the double mutants Gln35Ser/Asn64Phe (Q35S/N64F) and Gln35Lys/Asn64Phe (Q35K/N64F) of hDHFR that unexpectedly show evidence for the binding of two molecules of TMP: one molecule that binds in the normal folate binding site and the second molecule that binds in a novel subpocket site such that the mutated residue Phe64 is involved in van der Waals contacts to the trimethoxyphenyl ring of the second TMP molecule. Kinetic data for the binding of TMP to hDHFR and pjDHFR reveal an 84-fold selectivity of TMP against pjDHFR (Ki 49 nM) compared to hDHFR (Ki 4093 nM). Two mutants that contain one substitution from pj- and one from the closely related Pneumocystis carinii DHFR (pcDHFR) (Q35K/N64F and Q35S/N64F) show Ki values of 593 and 617 nM, respectively; these Ki values are well above both the Ki for pjDHFR and are similar to pcDHFR (Q35K/N64F) and Q35S/N64F) (305 nM). These results suggest that active site residues 35 and 64 play key roles in determining selectivity for pneumocystis DHFR, but that other residues contribute to the unique binding of inhibitors to these enzymes. PMID:21684339
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Diminished but Not Abolished Effect of Two His351 Mutants of Anthrax Edema Factor in a Murine Model
Zhao, Taoran; Zhao, Xinghui; Liu, Ju; Meng, Yingying; Feng, Yingying; Fang, Ting; Zhang, Jinlong; Yang, Xiuxu; Li, Jianmin; Xu, Junjie; Chen, Wei
2016-01-01
Edema toxin (ET), which is composed of a potent adenylate cyclase (AC), edema factor (EF), and protective antigen (PA), is one of the major toxicity factors of Bacillus anthracis. In this study, we introduced mutations in full-length EF to generate alanine EF(H351A) and arginine EF(H351R) variants. In vitro activity analysis displayed that the adenylyl cyclase activity of both the mutants was significantly diminished compared with the wild-type EF. When the native and mutant toxins were administered subcutaneously in a mouse footpad edema model, severe acute swelling was evoked by wild-type ET, while the symptoms induced by mutant toxins were very minor. Systemic administration of these EF variants caused non-lethal hepatotoxicity. In addition, EF(H351R) exhibited slightly higher activity in causing more severe edema than EF(H351A). Our findings demonstrate that the toxicity of ET is not abolished by substitution of EF residue His351 by alanine or arginine. These results also indicate the potential of the mouse footpad edema model as a sensitive method for evaluating both ET toxicity and the efficacy of candidate therapeutic agents. PMID:26848687
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Genetic analysis and gene mapping of a low stigma exposed mutant gene by high-throughput sequencing.
Directory of Open Access Journals (Sweden)
Xiao Ma
Full Text Available Rice is one of the main food crops and several studies have examined the molecular mechanism of the exposure of the rice plant stigma. The improvement in the exposure of the stigma in female parent hybrid combinations can enhance the efficiency of hybrid breeding. In the present study, a mutant plant with low exposed stigma (lesr was discovered among the descendants of the indica thermo-sensitive sterile line 115S. The ES% rate of the mutant decreased by 70.64% compared with the wild type variety. The F2 population was established by genetic analysis considering the mutant as the female parent and the restorer line 93S as the male parent. The results indicated a normal F1 population, while a clear division was noted for the high and low exposed stigma groups, respectively. This process was possible only by a ES of 25% in the F2 population. This was in agreement with the ratio of 3:1, which indicated that the mutant was controlled by a recessive main-effect QTL locus, temporarily named as LESR. Genome-wide comparison of the SNP profiles between the early, high and low production bulks were constructed from F2 plants using bulked segregant analysis in combination with high-throughput sequencing technology. The results demonstrated that the candidate loci was located on the chromosome 10 of the rice. Following screening of the recombinant rice plants with newly developed molecular markers, the genetic region was narrowed down to 0.25 Mb. This region was flanked by InDel-2 and InDel-2 at the physical location from 13.69 to 13.94 Mb. Within this region, 7 genes indicated base differences between parents. A total of 2 genes exhibited differences at the coding region and upstream of the coding region, respectively. The present study aimed to further clone the LESR gene, verify its function and identify the stigma variation.
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Probiotic features of Lactobacillus plantarum mutant strains.
Bove, Pasquale; Gallone, Anna; Russo, Pasquale; Capozzi, Vittorio; Albenzio, Marzia; Spano, Giuseppe; Fiocco, Daniela
2012-10-01
In this study, the probiotic potential of Lactobacillus plantarum wild-type and derivative mutant strains was investigated. Bacterial survival was evaluated in an in vitro system, simulating the transit along the human oro-gastro-intestinal tract. Interaction with human gut epithelial cells was studied by assessing bacterial adhesive ability to Caco-2 cells and induction of genes involved in innate immunity. L. plantarum strains were resistant to the combined stress at the various steps of the simulated gastrointestinal tract. Major decreases in the viability of L. plantarum cells were observed mainly under drastic acidic conditions (pH ≤ 2.0) of the gastric compartment. Abiotic stresses associated to small intestine poorly affected bacterial viability. All the bacterial strains significantly adhered to Caco-2 cells, with the ΔctsR mutant strain exhibiting the highest adhesion. Induction of immune-related genes resulted higher upon incubation with heat-inactivated bacteria rather than with live ones. For specific genes, a differential transcriptional pattern was observed upon stimulation with different L. plantarum strains, evidencing a possible role of the knocked out bacterial genes in the modulation of host cell response. In particular, cells from Δhsp18.55 and ΔftsH mutants strongly triggered immune defence genes. Our study highlights the relevance of microbial genetic background in host-probiotic interaction and might contribute to identify candidate bacterial genes and molecules involved in probiosis.
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Methods of producing protoporphyrin IX and bacterial mutants therefor
Zhou, Jizhong; Qiu, Dongru; He, Zhili; Xie, Ming
2016-03-01
The presently disclosed inventive concepts are directed in certain embodiments to a method of producing protoporphyrin IX by (1) cultivating a strain of Shewanella bacteria in a culture medium under conditions suitable for growth thereof, and (2) recovering the protoporphyrin IX from the culture medium. The strain of Shewanella bacteria comprises at least one mutant hemH gene which is incapable of normal expression, thereby causing an accumulation of protoporphyrin IX. In certain embodiments of the method, the strain of Shewanella bacteria is a strain of S. loihica, and more specifically may be S. loihica PV-4. In certain embodiments, the mutant hemH gene of the strain of Shewanella bacteria may be a mutant of shew_2229 and/or of shew_1140. In other embodiments, the presently disclosed inventive concepts are directed to mutant strains of Shewanella bacteria having at least one mutant hemH gene which is incapable of normal expression, thereby causing an accumulation of protoporphyrin IX during cultivation of the bacteria. In certain embodiments the strain of Shewanella bacteria is a strain of S. loihica, and more specifically may be S. loihica PV-4. In certain embodiments, the mutant hemH gene of the strain of Shewanella bacteria may be a mutant of shew_2229 and/or shew_1140.
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Directory of Open Access Journals (Sweden)
Deqiu Dai
2018-06-01
Full Text Available We reported the petrography and mineral chemistry of 184 equilibrated ordinary chondrites collected from Grove Mountains, Antarctica. The chemical-petrographic types and shock metamorphism degrees of these chondrites were assigned. They were classified into 46 H groups (22 H4, 20 H5, and four H6, 133 L groups (eight L4, 75 L5, and 50 L6, and five LL groups (four LL4 and one LL5. Some of these chondrites could be paired; however, both H and L group meteorites were affected. Further studies such as terrestrial ages and thermal luminescence are required in order to confirm the pairings. The relative abundances of H, L, and LL are different in Grove Mountain meteorites, when compared to those in Transcontinental Ridge meteorites. Based on the shock effects, the shock metamorphism degrees of these chondrites were assigned. Compared to previous studies, the heavily shocked samples of S4 and S5 have a higher fraction (59 out of 184 in Grove Mountain ordinary chondrites. The L group (54 out of 59 is the dominant chemical group in the heavily shocked chondrites, except for five meteorites which belong to the H group. The shock metamorphism degrees of the H and L groups are distinct, which may indicate different surface properties in their parent bodies. In addition, the petrologic types and shock degrees are probably closely related, with the most heavily shocked chondrites observed in types 5 and 6.
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Energy Technology Data Exchange (ETDEWEB)
Ely, Roger L.; Chaplen, Frank W.R.
2014-03-11
This project used the cyanobacterial species Synechocystis PCC 6803 to pursue two lines of inquiry, with each line addressing one of the two main factors affecting hydrogen (H2) production in Synechocystis PCC 6803: NADPH availability and O2 sensitivity. H2 production in Synechocystis PCC 6803 requires a very high NADPH:NADP+ ratio, that is, the NADP pool must be highly reduced, which can be problematic because several metabolic pathways potentially can act to raise or lower NADPH levels. Also, though the [NiFe]-hydrogenase in PCC 6803 is constitutively expressed, it is reversibly inactivated at very low O2 concentrations. Largely because of this O2 sensitivity and the requirement for high NADPH levels, a major portion of overall H2 production occurs under anoxic conditions in the dark, supported by breakdown of glycogen or other organic substrates accumulated during photosynthesis. Also, other factors, such as N or S limitation, pH changes, presence of other substances, or deletion of particular respiratory components, can affect light or dark H2 production. Therefore, in the first line of inquiry, under a number of culture conditions with wild type (WT) Synechocystis PCC 6803 cells and a mutant with impaired type I NADPH-dehydrogenase (NDH-1) function, we used H2 production profiling and metabolic flux analysis, with and without specific inhibitors, to examine systematically the pathways involved in light and dark H2 production. Results from this work provided rational bases for metabolic engineering to maximize photobiological H2 production on a 24-hour basis. In the second line of inquiry, we used site-directed mutagenesis to create mutants with hydrogenase enzymes exhibiting greater O2 tolerance. The research addressed the following four tasks: 1. Evaluate the effects of various culture conditions (N, S, or P limitation; light/dark; pH; exogenous organic carbon) on H2 production profiles of WT cells and an NDH-1 mutant; 2. Conduct metabolic flux analyses for
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Isolation of new gravitropic mutants under hypergravity conditions
Directory of Open Access Journals (Sweden)
Akiko Mori
2016-09-01
Full Text Available Forward genetics is a powerful approach used to link genotypes and phenotypes, and mutant screening/analysis has provided deep insights into many aspects of plant physiology. Gravitropism is a tropistic response in plants, in which hypocotyls and stems sense the direction of gravity and grow upwards. Previous studies of gravitropic mutants have suggested that shoot endodermal cells in Arabidopsis stems and hypocotyls are capable of sensing gravity (i.e., statocytes. In the present study, we report a new screening system using hypergravity conditions to isolate enhancers of gravitropism mutants, and we also describe a rapid and efficient genome mapping method, using Next-Generation Sequencing (NGS and Single Nucleotide Polymorphism (SNP-based markers. Using the endodermal-amyloplast less 1 (eal1 mutant, which exhibits defective development of endodermal cells and gravitropism, we found that hypergravity (10 g restored the reduced gravity responsiveness in eal1 hypocotyls and could, therefore, be used to obtain mutants with further reduction in gravitropism in the eal1 background. Using the new screening system, we successfully isolated six ene (enhancer of eal1 mutants that exhibited little or no gravitropism under hypergravity conditions, and using NGS and map-based cloning with SNP markers, we narrowed down the potential causative genes, which revealed a new genetic network for shoot gravitropism in Arabidopsis.
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Molybdenum x-ray absorption studies of the mutant Kp nifV of nitrogenase MO-FE protein
International Nuclear Information System (INIS)
Eidsness, M.K.; Smith, B.E.; Flood, A.C.; Garner, C.D.; Cramer, S.P.
1985-01-01
The nifV mutant nitrogenase enzyme of Klebsiella pheumoniae exhibits altered substrate reducing activity. This nitrogenase mutant cannot fix N 2 in vivo but can reduce C 2 H 2 to C 2 H 4 . The nifV mutant enzyme differs further from the wild-type enzyme by CO inhibition of its H 2 evolution activity, up to 80%. The NifV - phenotype (NifV - Kp1) has been shown to be associated with the iron-molybdenum cofactor (FeMoco) in the Mo Fe protein which is generally accepted as the site for substrate reduction. An X-Ray absorption study of the Mo site in this mutant may reveal a difference in its FeMoco structure. The authors report here a comparison of Mo X-Ray absorption data from the nitrogenase enzymes of the wild-type and NifV - strains in three different forms: (1) as isolated, (2) dye-oxidized, and (3) fixing enzyme systems. Mo edge structure of NifV - Kp1 and wild-type enzymes are nearly identical. Small shifts to higher energies are observed in the oxidized and fixing states. Mo EXAFS of NifV - Kp1 and wild-type in the ''as isolated'' state appear indistinguishable. Curve fitting results best describe the molybdenum in FeMoco as bound by 4-5 S atoms at 2.36 A ,3 Fe atoms at 2.69 A, and 0-2 O(N) atoms at 2.19 A. The spectral similarity of these results concerning the nifV mutant FeMoco structure is discussed
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A temperature-sensitive winter wheat chlorophyll mutant derived from space mutagenesis
International Nuclear Information System (INIS)
Zhao Hongbin; Guo Huijun; Zhao Linshu; Gu Jiayu; Zhao Shirong; Li Junhui; Liu Luxiang
2010-01-01
A temperature-sensitive winter wheat (Triticum aestivum L.) chlorophyll mutant Mt18, induced by spaceflight mutagenesis, was studied on agronomic traits, ultrastructure of chloroplast and photosynthesis characteristics. The leaf color of the mutant Mt18 showed changes from green to albino and back to green during the whole growth period. Plant height, productive tillers, spike length, grains and grain weight per plant, and 1000-grain weight of the mutant were lower than those of the wild type. The ultrastructural observation showed that no significant difference was found between the mutant and the wild type during prior albino stage, however, at the albino stage the number of granum-thylakoids and grana lamellae became fewer or completely disappeared, but the strom-thylakoid was obviously visible. After turning green,the structure of most chloroplasts recovered to normal, but number of chloroplast was still lower than that of the wild type. When exposed to photosynthetic active radiation (PAR) of 110 μmol·m -2 ·s -1 , the non-photochemical quenching (NPQ) of mutant was significantly lower than that of the wild type, and the non-regulated energy dissipation (Y NO ) was significantly higher than that of the wild type, while the change of the maximum photosystem II quantum yield (F v /F m ), potential activity of photosystem II (F v /F o ), photochemical quenching (q P ), effective quantum yield (Y PSI I) and regulated non-photochemical energy dissipation (Y NPQ ) were different at various stages. In addition, the differences of the electron transport rate (ETR), photochemical quenching (q P ), and effective quantum yield (Y PSI I) between mutant and wild type varied under different PAR conditions. It was concluded that with the change of chloroplast ultrastructure, the leaf color and photosynthesis of the wheat mutant Mt18 change correspondingly. The chloroplast ultrastructure was obviously different from that of wild type, and the photosynthetic efficiency
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Radiological environmental monitoring with LiF-700-H dosemeters
International Nuclear Information System (INIS)
Scarnichia, E.; Levanon, I.; Andres, P.; Miani, C.; Ramirez, S.
2011-10-01
Since 2008 a radiological environmental monitoring with LiF-700-H has been carried out as a result of increasing the Ra-6 research reactor core power. The information obtained is used to evaluate and to quantify analytically the air kerma rate, the fading and the associated uncertainty by developing software tools (deconvolution and uncertainty algorithms). LiF-700-H dosemeters were chosen because of their high sensitivity to low air kerma rates. They show a very good stability and a negligible fading for two-month working periods. The air kerma rate detection limit (based on the 3σ criterion) during these working periods is about 0.4 n Gy/h. Air kerma rates of about 70 n Gy/h are measured with this detection limit. Following the Nist guidelines, an algorithm was developed in order to find the associated uncertainty. It considers several aspects, such as the source activity decay, distance source-dosemeter during the calibration procedure, irradiation time, calibration factor, dosemeter readout, dosemeter sensitivity, TLD reader stability and fading. The associated uncertainty is found to be about 25% for a 95% confidence interval (k = 2.025), which can be considered acceptable when taking into account the very low air kerma rates estimated. The LiF-700-H response to different energies and its relationship with climate changes over the calendar year are planned as future tasks. (Author)
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En dialog för hållbarhet : En studie om statliga företags intressentdialog i praktiken
Danielsson, Josefin; Ulleryd, Ida
2016-01-01
Kraven på att organisationer ska ta ett socialt, miljömässigt och ekonomiskt ansvar har ökat i och med den ökade medvetenhet om hur olika verksamheter påverkar såväl miljö som samhälle. Det huvudsakliga verktyget för företag att kommunicera sitt ansvars- och hållbarhetsarbete är genom hållbarhetsredovisningen, där Global Reporting Initiatives (GRI) riktlinjer är den mest använda standarden vid upprättandet av en hållbarhetsredovisning. En av de grundläggande principerna i GRI är att företagen...
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International Nuclear Information System (INIS)
Naidenova, N.; Vassilevska-Ivanova, R.; Tcekova, Z.
2003-01-01
Two foliar pea (Pisum sativum L.) mutants characterized by reduced leaf size (2/978) and orange leaf (2/1409 M) were established. Both mutants were described morphologically and their productivity potential , pollen viability and inheritance of the mutant traits were evaluated. The mutant 2/978 was identified after irradiation of dry seeds from cv Borek with 15 Gy fast neutrons and was related to the leaf mutation 'rogue'. Reciprocal crosses between mutant 2/978 and cv Borel were executed, and F 1 and F 2 generations were analyzed. The altered leaf trait was presented in all F 1 plants suggesting a dominant character. F 2 segregation data indicated that the trait was controlled by a single dominant gene. The mutant 2/1409M originated from the mutant 2/978 after irradiation with 50 Gy γ-rays. The main mutant's phenotypic characteristic was the orange-yellow coloration of leaves and plants. After of series of crosses it was established that induced chlorophyll mutation is monogenic, recessive and both mutant traits are independently inherited. Two mutants could be used as appropriate plant material for genetic and biological investigations
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21 CFR 184.1804 - Sodium potassium tartrate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sodium potassium tartrate. 184.1804 Section 184.1804 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... has a cooling saline taste. It is obtained as a byproduct of wine manufacture. (b) The ingredient...
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Saturation behavior of irradiation hardening in F82H irradiated in the HFIR
Energy Technology Data Exchange (ETDEWEB)
Hirose, T. [Blanket Engineering Group, Japan Atomic Energy Agency, Naka, Ibaraki (Japan); Shiba, K.; Tanigawa, H.; Ando, M. [Japan Atomic Energy Agency, Tokai-mura, Naga-gun, Ibaraki-ken (Japan); Klueh, R.L. [Oak Ridge National Laboratory, TN (United States); Stoller, R. [ORNL - Oak Ridge National Laboratory, Materials Science and Technology Div., Oak Ridge, AK TN (United States)
2007-07-01
Full text of publication follows: Post irradiation tensile tests on reduced activation ferritic/martensitic steel, F82H have been conducted over the past two decades using Japan Materials Testing Reactor (JMTR) of JAEA, and Fast Flux Testing Facility (FFTF) of PNNL and High Flux Isotope Reactor (HFIR) of ORNL, USA, under Japan/US collaboration programs. According to these results, F82H does not demonstrate irradiation hardening above 673 K up to 60 dpa. The current study has been concentrated on hardening behavior at temperature around 573 K. A series of low temperature irradiation experiment has been conducted at the HFIR under the international collaborative research between JAEA/US-DOE. In this collaboration, the irradiation condition is precisely controlled by the well matured capsule designing and instrumentation. This paper summarizes recent results of the irradiation experiments focused on F82H and its modified steels compared with the irradiation properties database on F82H. Post irradiation tensile tests have been conducted on the F82H and its modified steels irradiated at 573 K and the dose level was up to 25 dpa. According to these results, irradiation hardening of F82H is saturated by 9 dpa and the as-irradiated 0.2 % proof stress is less than 1 GPa at ambient temperature. The deterioration of total elongation was also saturated by 9 dpa irradiation. The ductility of some modified steels which showed larger total elongation than that of F82H before irradiation become the same level as that of standard F82H steel after irradiation, even though its magnitude of irradiation hardening is smaller than that of F82H. This suggests that the more ductile steel demonstrates the more ductility loss at this temperature, regardless to the hardening level. The difference in ductility loss behavior between various tensile specimens will be discussed as the ductility could depend on the specimen dimension. (authors)
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Prevention of GABA reduction during dough fermentation using a baker's yeast dal81 mutant.
Ando, Akira; Nakamura, Toshihide
2016-10-01
γ-Aminobutyric acid (GABA) is consumed by yeasts during fermentation. To prevent GABA reduction in bread dough, a baker's yeast mutant AY77 deficient in GABA assimilation was characterized and utilized for wheat dough fermentation. An amber mutation in the DAL81 gene, which codes for a positive regulator of multiple nitrogen degradation pathways, was found in the AY77 strain. The qPCR analyses of genes involved in nitrogen utilization showed that transcriptional levels of the UGA1 and DUR3 genes encoding GABA transaminase and urea transporter, respectively, are severely decreased in the AY77 cells. The AY77 strain cultivated by fed-batch culture using cane molasses exhibited inferior gas production during dough fermentation compared to that of wild-type strain AY13. However, when fed with molasses containing 0.5% ammonium sulfate, the mutant strain exhibited gas production comparable to that of the AY13 strain. In contrast to the AY13 strain, which completely consumed GABA in dough within 5 h, the AY77 strain consumed no GABA under either culture condition. Dough fermentation with the dal81 mutant strain should be useful for suppression of GABA reduction in breads. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.
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The role of RNase H2 in processing ribonucleotides incorporated during DNA replication.
Williams, Jessica S; Gehle, Daniel B; Kunkel, Thomas A
2017-05-01
Saccharomyces cerevisiae RNase H2 resolves RNA-DNA hybrids formed during transcription and it incises DNA at single ribonucleotides incorporated during nuclear DNA replication. To distinguish between the roles of these two activities in maintenance of genome stability, here we investigate the phenotypes of a mutant of yeast RNase H2 (rnh201-RED; ribonucleotide excision defective) that retains activity on RNA-DNA hybrids but is unable to cleave single ribonucleotides that are stably incorporated into the genome. The rnh201-RED mutant was expressed in wild type yeast or in a strain that also encodes a mutant allele of DNA polymerase ε (pol2-M644G) that enhances ribonucleotide incorporation during DNA replication. Similar to a strain that completely lacks RNase H2 (rnh201Δ), the pol2-M644G rnh201-RED strain exhibits replication stress and checkpoint activation. Moreover, like its null mutant counterpart, the double mutant pol2-M644G rnh201-RED strain and the single mutant rnh201-RED strain delete 2-5 base pairs in repetitive sequences at a high rate that is topoisomerase 1-dependent. The results highlight an important role for RNase H2 in maintaining genome integrity by removing single ribonucleotides incorporated during DNA replication. Published by Elsevier B.V.
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Gamma-ray induced mutants in castor (Ricinus communis L.)
International Nuclear Information System (INIS)
Janila, P.; Ashok Kumar, A.; Rajashekar Reddy, N.; Hemalatha, V.
2007-01-01
We report isolation of three recessive mutants in castor using dry seed irradiation with gamma rays. The crinkled leaf mutant (crf) was identified in K-55-112 M2 family and leafy mutant (lea) in H-55-577 M2 family; both are recessive lethal and thus maintained as heterozygotes. The cri mutant has highly wrinkled leaves resembling finger millet head and failed to enter reproductive phase, consequently did not produce seeds. The number of leaf lobes is reduced in lea mutant and though it produced spikes, the male and female flowers are converted to leafy appendages. The third mutant, fused (Ius) stem identified in H-55-617 M2 family is a recessive mutant. The branches of which are fused at the base and though each branch terminates in to monoceous spike like normal plant, the spike is highly condensed. The three mutants under report are valuable genetic stocks for development of linkage maps in castor, which is at infancy. (author)
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21 CFR 184.1077 - Potassium acid tartrate.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Potassium acid tartrate. 184.1077 Section 184.1077 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD... a white, crystalline powder. It has a pleasant, acid taste. It is obtained as a byproduct of wine...
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Double-tuned radiofrequency coil for (19)F and (1)H imaging.
Otake, Yosuke; Soutome, Yoshihisa; Hirata, Koji; Ochi, Hisaaki; Bito, Yoshitaka
2014-01-01
We developed a double-tuned radiofrequency (RF) coil using a novel circuit method to double tune for fluorine-19 (19F) and 1H magnetic resonance imaging, whose frequencies are very close to each other. The RF coil consists of 3 parallel-connected series inductor capacitor circuits. A computer simulation for our double-tuned RF coil with a phantom demonstrated that the coil has tuned resonant frequency and high sensitivity for both 19F and 1H. Drug distribution was visualized at 7 tesla using this RF coil and a rat administered perfluoro 15-crown-5-ether emulsion. The double-tune RF coil we developed may be a powerful tool for 19F and 1H imaging.
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Interface strength measurement of tungsten coatings on F82H substrates
International Nuclear Information System (INIS)
Kim, Hyoungil; El-Awady, Jaafar; Gupta, Vijay; Ghoniem, Nasr; Sharafat, Shahram
2009-01-01
In the current work, hot isostatic pressing is adopted to deposit tungsten coatings on F82H substrates. The interface strength of the W/F82H samples is measured using the Laser Spallation technique and the microstructure is analyzed to determine the strength of the coating. Finally, the failure mechanisms of the hot isostatic pressing versus vacuum plasma spraying tungsten coatings and their different failure strengths are compared. It is concluded that the hot isostatic pressing process ensures a good adhesion for the W/F82H interface while the vacuum plasma spraying process results in relatively lower failure strength for the W-coating itself due to the high porosity in the coating.
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Analysis of Organic Samples from the 5-H and 3-F Pump Tanks and Waste Tank 38H
International Nuclear Information System (INIS)
Swingle, R.F. II
1999-01-01
Analyses for organic materials in aqueous and surface floating samples taken from the 5-H Pump Tank and Waste Tank 38H and in vapor samples taken from the 5-H and 3-F Pump Tanks have been completed. The results indicate that the concentration of organic materials is extremely low in all samples. This report documents the development of sampling and analysis techniques for this sampling as well as the results of the analyses of vapor samples pulled from Pump Tanks 5-H and 3-F and liquid samples pulled from Waste Tank 38H and Pump Tank 5-H
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Temperature sensitive riboflavin mutants of Penicillium vermiculatum Dangeard
International Nuclear Information System (INIS)
Mitra, J.; Chaudhari, K.L.
1974-01-01
Two temperature sensitive UV induced riboflavin mutants rib 1 and rib 6 have been physiologically and genetically characterized. The two mutants behave differently with regard to their temperature sensitivity. The rib 1 mutant exhibits a leaky growth in minimal medium between 15 0 C and 30 0 C but grows well when the medium is supplemented with riboflavin. At 35 0 C the growth response of the mutant is at its max. and at 40 0 C and below 15 0 C it ceases to grow. The rib 6 mutant which is red brown in colour shows wild type character at temp. below 25 0 C in minimal medium but requires riboflavin at 30 0 C and above. Heterokaryotic analysis revealed the nonallelic nature of the two temperature mutants. Genetic tests of allelic relationship between riboflavin markers by crossing were also done. (author)
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27 CFR 25.184 - Losses in transit.
2010-04-01
... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Losses in transit. 25.184... Ownership § 25.184 Losses in transit. (a) Liability for losses. The brewer is liable under the bond of the brewery to which beer is transferred for the tax on beer lost in transit. If the brewer reconsigns beer...
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Giuntini, Serena; Reason, Donald C; Granoff, Dan M
2012-01-01
Meningococcal vaccines containing factor H binding protein (fHbp) are in clinical development. fHbp binds human fH, which enables the meningococcus to resist complement-mediated bacteriolysis. Previously, we found that chimeric human IgG1 mouse anti-fHbp monoclonal antibodies (MAbs) had human complement-mediated bactericidal activity only if the MAb inhibited fH binding. Since IgG subclasses differ in their ability to activate complement, we investigated the role of human IgG subclasses on antibody functional activity. We constructed chimeric MAbs in which three different murine fHbp-specific binding domains were each paired with human IgG1, IgG2, or IgG3. Against a wild-type group B isolate, all three IgG3 MAbs, irrespective of their ability to inhibit fH binding, had bactericidal activity that was >5-fold higher than the respective IgG1 MAbs, while the IgG2 MAbs had the least activity. Against a mutant with increased fHbp expression, the anti-fHbp MAbs elicited greater C4b deposition (classical pathway) and greater bactericidal activity than against the wild-type strain, and the IgG1 MAbs had similar or greater activity than the respective IgG3 MAbs. The bactericidal activity against both wild-type and mutant strains also was dependent, in part, on activation of the alternative complement pathway. Thus, at lower epitope density in the wild-type strain, the IgG3 anti-fHbp MAbs had the greatest bactericidal activity. At a higher epitope density in the mutant, the IgG1 MAbs had similar or greater bactericidal activity than the IgG3 MAbs, and the activity was less dependent on the inhibition of fH binding than at a lower epitope density.
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19 CFR 191.184 - Merchandise transferred from continuous Customs custody.
2010-04-01
... 19 Customs Duties 2 2010-04-01 2010-04-01 false Merchandise transferred from continuous Customs custody. 191.184 Section 191.184 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND... From Customs Territory § 191.184 Merchandise transferred from continuous Customs custody. (a) Procedure...
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Comportamiento productivo en híbridos de jitomate y sus respectivas poblaciones F2
Directory of Open Access Journals (Sweden)
J. Martínez-Solís
2005-01-01
Full Text Available Debido a que el precio de semilla híbrida de jitomate es muy alto, muchos agricultores utilizan generaciones F2 en parcelas comerciales. Por esta razón con el propósito de evaluar el comportamiento entre genotipos de jitomate F1 y F2, 37 híbridos y sus respectivas generaciones F2 fueron establecidos en invernadero. La mayoría de las generaciones F2 presentaron menor peso total de fruto por planta y número de frutos por planta con respecto a sus F1; sin embargo, el efecto fue estadísticamente significativo sólo en dos genotipos tipo bola de hábito de crecimiento determinado y cuatro genotipos tipo bola de hábito de crecimiento indeterminado, los cuales abatieron su producción de fruto de F2 en al menos 20 %
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Lasho, Terra L; Pardanani, Animesh; McClure, Rebecca F; Mesa, Ruben A; Levine, Ross L; Gilliland, D Gary; Tefferi, Ayalew
2006-12-01
MPLW515L/K and JAK2V617F can co-exist in myelofibrosis with myeloid metaplasia (MMM). The chronology of clonal emergence was studied in three such cases using serially stored bone marrow. At diagnosis, a major MPL515 mutant clone was accompanied by a minor JAK2V617F clone in all three instances. At 25 time points over a period of 4-8 years, allele burden fluctuated but remained high for MPLW515L/K and low for JAK2V617F. We conclude that MPLW515L/K and JAK2V617F are both early events in MMM and allele burden, rather than the mere presence of these mutations, might be relevant to phenotypic variation in myeloproliferative disorders.
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Giannelli, Serena G; Luoni, Mirko; Castoldi, Valerio; Massimino, Luca; Cabassi, Tommaso; Angeloni, Debora; Demontis, Gian Carlo; Leocani, Letizia; Andreazzoli, Massimiliano; Broccoli, Vania
2018-03-01
P23H is the most common mutation in the RHODOPSIN (RHO) gene leading to a dominant form of retinitis pigmentosa (RP), a rod photoreceptor degeneration that invariably causes vision loss. Specific disruption of the disease P23H RHO mutant while preserving the wild-type (WT) functional allele would be an invaluable therapy for this disease. However, various technologies tested in the past failed to achieve effective changes and consequently therapeutic benefits. We validated a CRISPR/Cas9 strategy to specifically inactivate the P23H RHO mutant, while preserving the WT allele in vitro. We, then, translated this approach in vivo by delivering the CRISPR/Cas9 components in murine Rho+/P23H mutant retinae. Targeted retinae presented a high rate of cleavage in the P23H but not WT Rho allele. This gene manipulation was sufficient to slow photoreceptor degeneration and improve retinal functions. To improve the translational potential of our approach, we tested intravitreal delivery of this system by means of adeno-associated viruses (AAVs). To this purpose, the employment of the AAV9-PHP.B resulted the most effective in disrupting the P23H Rho mutant. Finally, this approach was translated successfully in human cells engineered with the homozygous P23H RHO gene mutation. Overall, this is a significant proof-of-concept that gene allele specific targeting by CRISPR/Cas9 technology is specific and efficient and represents an unprecedented tool for treating RP and more broadly dominant genetic human disorders affecting the eye, as well as other tissues.
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Kuo, Heng-Lan; Lien, Jin-Cherng; Chung, Ching-Hu; Chang, Chien-Hsin; Lo, Shyh-Chyi; Tsai, I-Chun; Peng, Hui-Chin; Kuo, Sheng-Chu; Huang, Tur-Fu
2010-06-01
The established antiplatelet and anticoagulant agents show beneficial effects in the treatment of thromboembolic diseases; however, these drugs still have considerable limitations. The effects of NP-184, a synthetic compound, on platelet functions, plasma coagulant activity, and mesenteric venule thrombosis in mice were investigated. NP-184 concentration-dependently inhibited the human platelet aggregation induced by collagen, arachidonic acid (AA), and U46619, a thromboxane (TX)A(2) mimic, with IC(50) values of 4.5 +/- 0.2, 3.9 +/- 0.1, and 9.3 +/- 0.5 microM, respectively. Moreover, NP-184 concentration-dependently suppressed TXA(2) formations caused by collagen and AA. In exploring effects of NP-184 on enzymes involved in TXA(2) synthesis, we found that NP-184 selectively inhibited TXA(2) synthase activity with an IC(50) value of 4.3 +/- 0.2 microM. Furthermore, NP-184 produced a right shift of the concentration-response curve of U46619, indicating a competitive antagonism on TXA(2)/prostaglandin H(2) receptor. Intriguingly, NP-184 also caused a concentration-dependent prolongation of the activated partial thromboplastin time (aPTT) with no changes in the prothrombin and thrombin time, indicating that it selectively impairs the intrinsic coagulation pathway. Oral administration of NP-184 significantly inhibited thrombus formation of the irradiated mesenteric venules in fluorescein sodium-treated mice without affecting the bleeding time induced by tail transection. However, after oral administration, NP-184 inhibited the ex vivo mouse platelet aggregation triggered by collagen and U46619 and also prolonged aPTT. Taken together, the dual antiplatelet and anticoagulant activities of NP-184 may have therapeutic potential as an oral antithrombotic agent in the treatment of thromboembolic disorders.
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F. H. C h . J othmann , De Bijbel in de loop der eeuwen. Kampen, J ...
African Journals Online (AJOL)
Test
F. H. C h . J othmann , De Bijbel in de loop der eeuwen. Kampen, J . H. Kok. N.V., 1954, 43 bis. Prys 95 cent. Hierdie werkie geskryf vir gebruik op skool, kategesasie en jeug- vereniging is besonder smakelik versorg met uiters insiggewende illu- strasies oor die geskiedenis van die Bybel as geskrif en boek. Die stof is.
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Directory of Open Access Journals (Sweden)
Rolando Pajon
2011-09-01
Full Text Available Factor H binding protein (fHbp is an important antigen for vaccines against meningococcal serogroup B disease. The protein binds human factor H (fH, which enables the bacteria to resist serum bactericidal activity. Little is known about the vaccine-potential of fHbp for control of meningococcal epidemics in Africa, which typically are caused by non-group B strains.We investigated genes encoding fHbp in 106 serogroup A, W-135 and X case isolates from 17 African countries. We determined complement-mediated bactericidal activity of antisera from mice immunized with recombinant fHbp vaccines, or a prototype native outer membrane vesicle (NOMV vaccine from a serogroup B mutant strain with over-expressed fHbp. Eighty-six of the isolates (81% had one of four prevalent fHbp sequence variants, ID 4/5 (serogroup A isolates, 9 (W-135, or 74 (X in variant group 1, or ID 22/23 (W-135 in variant group 2. More than one-third of serogroup A isolates and two-thirds of W-135 isolates tested had low fHbp expression while all X isolates tested had intermediate or high expression. Antisera to the recombinant fHbp vaccines were generally bactericidal only against isolates with fHbp sequence variants that closely matched the respective vaccine ID. Low fHbp expression also contributed to resistance to anti-fHbp bactericidal activity. In contrast to the recombinant vaccines, the NOMV fHbp ID 1 vaccine elicited broad anti-fHbp bactericidal activity, and the antibodies had greater ability to inhibit binding of fH to fHbp than antibodies elicited by the control recombinant fHbp ID 1 vaccine.NOMV vaccines from mutants with increased fHbp expression elicit an antibody repertoire with greater bactericidal activity than recombinant fHbp vaccines. NOMV vaccines are promising for prevention of meningococcal disease in Africa and could be used to supplement coverage conferred by a serogroup A polysaccharide-protein conjugate vaccine recently introduced in some sub
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Thermodynamics and performance of the Mg-H-F system for thermochemical energy storage applications.
Tortoza, Mariana S; Humphries, Terry D; Sheppard, Drew A; Paskevicius, Mark; Rowles, Matthew R; Sofianos, M Veronica; Aguey-Zinsou, Kondo-Francois; Buckley, Craig E
2018-01-24
Magnesium hydride (MgH 2 ) is a hydrogen storage material that operates at temperatures above 300 °C. Unfortunately, magnesium sintering occurs above 420 °C, inhibiting its application as a thermal energy storage material. In this study, the substitution of fluorine for hydrogen in MgH 2 to form a range of Mg(H x F 1-x ) 2 (x = 1, 0.95, 0.85, 0.70, 0.50, 0) composites has been utilised to thermodynamically stabilise the material, so it can be used as a thermochemical energy storage material that can replace molten salts in concentrating solar thermal plants. These materials have been studied by in situ synchrotron X-ray diffraction, differential scanning calorimetry, thermogravimetric analysis, temperature-programmed-desorption mass spectrometry and Pressure-Composition-Isothermal (PCI) analysis. Thermal analysis has determined that the thermal stability of Mg-H-F solid solutions increases proportionally with fluorine content, with Mg(H 0.85 F 0.15 ) 2 having a maximum rate of H 2 desorption at 434 °C, with a practical hydrogen capacity of 4.6 ± 0.2 wt% H 2 (theoretical 5.4 wt% H 2 ). An extremely stable Mg(H 0.43 F 0.57 ) 2 phase is formed upon the decomposition of each Mg-H-F composition of which the remaining H 2 is not released until above 505 °C. PCI measurements of Mg(H 0.85 F 0.15 ) 2 have determined the enthalpy (ΔH des ) to be 73.6 ± 0.2 kJ mol -1 H 2 and entropy (ΔS des ) to be 131.2 ± 0.2 J K -1 mol -1 H 2 , which is slightly lower than MgH 2 with ΔH des of 74.06 kJ mol -1 H 2 and ΔS des = 133.4 J K -1 mol -1 H 2 . Cycling studies of Mg(H 0.85 F 0.15 ) 2 over six absorption/desorption cycles between 425 and 480 °C show an increased usable cycling temperature of ∼80 °C compared to bulk MgH 2 , increasing the thermal operating temperatures for technological applications.
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Heard, Jeffrey J; Phung, Ivy; Potes, Mark I; Tamanoi, Fuyuhiko
2018-01-10
RHEB is a unique member of the RAS superfamily of small GTPases expressed in all tissues and conserved from yeast to humans. Early studies on RHEB indicated a possible RHEB-RAF interaction, but this has not been fully explored. Recent work on cancer genome databases has revealed a reoccurring mutation in RHEB at the Tyr35 position, and a recent study points to the oncogenic potential of this mutant that involves activation of RAF/MEK/ERK signaling. These developments prompted us to reassess the significance of RHEB effect on RAF, and to compare mutant and wild type RHEB. To study RHEB-RAF interaction, and the effect of the Y35N mutation on this interaction, we used transfection, immunoprecipitation, and Western blotting techniques. We generated cell lines stably expressing RHEB WT, RHEB Y35N, and KRAS G12V, and monitored cellular transforming properties through cell proliferation, anchorage independent growth, cell cycle analysis, and foci formation assays. We observe a strong interaction between RHEB and BRAF, but not with CRAF. This interaction is dependent on an intact RHEB effector domain and RHEB-GTP loading status. RHEB overexpression decreases RAF activation of the RAF/MEK/ERK pathway and RHEB knockdown results in an increase in RAF/MEK/ERK activation. RHEB Y35N mutation has decreased interaction with BRAF, and RHEB Y35N cells exhibit greater BRAF/CRAF heterodimerization resulting in increased RAF/MEK/ERK signaling. This leads to cancer transformation of RHEB Y35N stably expressing cell lines, similar to KRAS G12 V expressing cell lines. RHEB interaction with BRAF is crucial for inhibiting RAF/MEK/ERK signaling. The RHEB Y35N mutant sustains RAF/MEK/ERK signaling due to a decreased interaction with BRAF, leading to increased BRAF/CRAF heterodimerization. RHEB Y35N expressing cells undergo cancer transformation due to decreased interaction between RHEB and BRAF resulting in overactive RAF/MEK/ERK signaling. Taken together with the previously established
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DEFF Research Database (Denmark)
Morris, V. M.; Gunning, A. P.; Faults, C. B.
2005-01-01
Atomic force microscopy has been used to investigate the complexes formed between high molecular weight amylose chains and Aspergillus niger glucoamylase mutants (E400Q and W52F), wild-type A. niger starch binding domains (SBDS), and mutant SBDs (W563K and W590K) lacking either of the two starch...
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RECEPTOR POTENTIAL AND LIGHT-INDUCED MITOCHONDRIAL ACTIVATION IN BLOWFLY PHOTORECEPTOR MUTANTS
MOJET, MH; TINBERGEN, J; STAVENGA, DG
1991-01-01
1. Simultaneous measurements of the receptor potential and the light-induced mitochondrial activation were performed in white-eyed blowflies Calliphora vicina, mutant chalky, and Lucilia cuprina, mutants w(F) and w'nss. The intensity dependence and the temporal dynamics were investigated. 2. The
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$\\alpha$-decay study of $^{182,184}$Tl
Van Beveren, C; Barzakh, A E; Cocolios, T E; de Groote, R P; Fedorov, D; Fedosseev, V N; Ferrer, R; Ghys, L; Huyse, M; Köster, U; Lane, J; Liberati, V; Lynch, K M; Marsh, B A; Molkanov, P L; Procter, T J; Rapisarda, E; Sandhu, K; Seliverstov, M D; Van Duppen, P; Venhart, M; Veselský, M
2016-01-01
α -decay spectroscopy of 182,184 Tl has been performed at the CERN isotope separator on-line ( ISOLDE ) facility. New fi ne-structure α decays have been observed for both isotopes. α -decay branching ratios of 0.089 ( 19 ) %, 0.047 ( 6 ) % and 1.22 ( 30 ) % have been deduced for the ( 10 − ) , ( 7 + ) and ( 2 − ) states respectively in 184 Tl and a lower limit of 0.49% for the α -decay branching ratio of 182 Tl. A new half-life of 9.5 ( 2 ) s for the ( 2 − ) state in 184 Tl and 1.9 ( 1 ) s for the low-spin state in 182 Tl has been deduced. Using α – γ coincidence analysis, multiple γ rays were observed de-exciting levels in 178,18 0 Au fed by 182,184 Tl α decays. The γ transitions connecting these low-lying states in 178,18 0 Au are essential to sort the data and possibly identify bands from in- beam studies in these isotopes. Owing to the complex fi ne-structure α decays and limited knowledge about the structure of the daughter nuclei, only partial level schemes could be constructed for bot...
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Energy Technology Data Exchange (ETDEWEB)
Kronenberg, A; Little, J B
1989-04-01
In order to characterize the nature of mutants induced by densely ionizing radiations at an autosomal locus, the authors have isolated a series of 99 thymidine kinase (tk) mutants of human TK6 lymphoblastoid cells iraadiated with either fast neutrons or accelerated argon ions. Individual muant clones were examined for alterations in their restriction fragment pattern after hybridization with a human cDNA probe for tk. A restriction fragment length polymorphism (RFLP) allowed identification of the active tk allele. Among the neutron-induced mutants, 34/52 exhibited loss of the previously active allele while 6/52 exhibited intragenic rearrangements. Among the argon-induced mutants 27/46 exhibited allele loses and 10/46 showed rearrangements within the tk locus. The remaining mutants had restriction patterns indistinguishable from the TK6 parent. Each of the mutant clones was further examined for structural alterations within the c-erbAl locus which has been localized to chromosome 17q11-q22, at some unknown distance from the human tk locus at chromosome 17q21-q22. A substantial proportion (54%) of tk mutants induced by densely ionizing radiation showed loss of the c-erb locus on the homologous chromosome, suggesting that the mutations involve large-scale genetic changes. (author). 51 refs.; 2 figs.; 6 tabs.
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Ito, Masahiro; Kim, Yun-Gi; Tsuji, Hirokazu; Takahashi, Takuya; Kiwaki, Mayumi; Nomoto, Koji; Danbara, Hirofumi; Okada, Nobuhiko
2014-01-01
Lactobacillus casei ATCC 27139 enhances host innate immunity, and the J1 phage-resistant mutants of this strain lose the activity. A transposon insertion mutant library of L. casei ATCC 27139 was constructed, and nine J1 phage-resistant mutants out of them were obtained. Cloning and sequencing analyses identified three independent genes that were disrupted by insertion of the transposon element: asnH, encoding asparagine synthetase, and dnaJ and dnaK, encoding the molecular chaperones DnaJ and DnaK, respectively. Using an in vivo mouse model of Listeria infection, only asnH mutant showed deficiency in their ability to enhance host innate immunity, and complementation of the mutation by introduction of the wild-type asnH in the mutant strain recovered the immuno-augmenting activity. AsnH protein exhibited asparagine synthetase activity when the lysozyme-treated cell wall extracts of L. casei ATCC 27139 was added as substrate. The asnH mutants lost the thick and rigid peptidoglycan features that are characteristic to the wild-type cells, indicating that AsnH of L. casei is involved in peptidoglycan biosynthesis. These results indicate that asnH is required for the construction of the peptidoglycan composition involved in the immune-activating capacity of L. casei ATCC 27139.
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Directory of Open Access Journals (Sweden)
Masahiro Ito
Full Text Available Lactobacillus casei ATCC 27139 enhances host innate immunity, and the J1 phage-resistant mutants of this strain lose the activity. A transposon insertion mutant library of L. casei ATCC 27139 was constructed, and nine J1 phage-resistant mutants out of them were obtained. Cloning and sequencing analyses identified three independent genes that were disrupted by insertion of the transposon element: asnH, encoding asparagine synthetase, and dnaJ and dnaK, encoding the molecular chaperones DnaJ and DnaK, respectively. Using an in vivo mouse model of Listeria infection, only asnH mutant showed deficiency in their ability to enhance host innate immunity, and complementation of the mutation by introduction of the wild-type asnH in the mutant strain recovered the immuno-augmenting activity. AsnH protein exhibited asparagine synthetase activity when the lysozyme-treated cell wall extracts of L. casei ATCC 27139 was added as substrate. The asnH mutants lost the thick and rigid peptidoglycan features that are characteristic to the wild-type cells, indicating that AsnH of L. casei is involved in peptidoglycan biosynthesis. These results indicate that asnH is required for the construction of the peptidoglycan composition involved in the immune-activating capacity of L. casei ATCC 27139.
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26 CFR 1.642(f)-1 - Amortization deductions.
2010-04-01
....642(f)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Estates, Trusts, and Beneficiaries § 1.642(f)-1 Amortization deductions. An estate... respect to qualified railroad rolling stock as defined in section 184(d), with respect to certified coal...
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Zhao, Ji-Li; Liu, Wei; Xie, Wan-Ying; Cao, Xu-Dong; Yuan, Li
2018-01-01
Tuberculosis (TB) caused by Mycobacterium tuberculosis (MTB) is one of the most common chronic infectious amphixenotic diseases worldwide. Prevention and control of TB are greatly difficult, due to the increase in drug-resistant TB, particularly multidrug-resistant TB. We speculated that there were some differences between drug-sensitive and drug-resistant MTB strains and that mazEF 3,6,9 toxin-antitoxin systems (TASs) were involved in MTB viability. This study aimed to investigate differences in viability, biofilm formation, and MazEF expression between drug-sensitive and drug-resistant MTB strains circulating in Xinjiang, China, and whether mazEF 3,6,9 TASs contribute to MTB viability under stress conditions. Growth profiles and biofilm-formation abilities of drug-sensitive, drug-resistant MTB strains and the control strain H37Rv were monitored. Using molecular biology experiments, the mRNA expression of the mazF 3, 6, and 9 toxin genes, the mazE 3, 6, and 9 antitoxin genes, and expression of the MazF9 protein were detected in the different MTB strains, H37RvΔ mazEF 3,6,9 mutants from the H37Rv parent strain were generated, and mutant viability was tested. Ex vivo culture analyses demonstrated that drug-resistant MTB strains exhibit higher survival rates than drug-sensitive strains and the control strain H37Rv. However, there was no statistical difference in biofilm-formation ability in the drug-sensitive, drug-resistant, and H37Rv strains. mazE 3,6 mRNA-expression levels were relatively reduced in the drug-sensitive and drug-resistant strains compared to H37Rv. Conversely, mazE 3,9 expression was increased in drug-sensitive strains compared to drug-resistant strains. Furthermore, compared with the H37Rv strain, mazF 3,6 expression was increased in drug-resistant strains, mazF 9 expression was increased in drug-sensitive strains, and mazF 9 exhibited reduced expression in drug-resistant strains compared with drug-sensitive strains. Protein expression of mazF9
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International Nuclear Information System (INIS)
Dantzig, A.H.; Slayman, C.W.; Adelberg, E.A.
1982-01-01
A spontaneous transport mutant of Chinese hamster ovary cells, CHY-1, was isolated by a combination of [ 3 H]proline suicide and replica plating. The mutant took up less tritium than the parent, resulting in a lower killing rate during storage. Transport by four separate amino acid transport systems (A, ASC, L, Ly+) was examined. The CHY-1 mutant exhibited normal uptake via the ASC, L, and Ly+ systems. By contrast, uptake of the most specific substrate of the A system, 2-(methylamino)-isobutyric acid, was significantly reduced at low, but not high, concentrations, due to a 3.5-fold increase in Km and a 1.5-fold increase in Vmax. Taken together, these data suggest that the CHY-1 mutation may be in the structural gene coding for the A transport protein. The tritium suicide procedure is discussed, and general equations are derived to predict the maximum storage time for the survival of one mutant cell and the optimum size of the cell population for maximum mutant enrichment
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21 CFR 184.1973 - Beeswax (yellow and white).
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Beeswax (yellow and white). 184.1973 Section 184.1973 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE...
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Rijal, Ramesh; Arhzaouy, Khalid; Strucksberg, Karl-Heinz; Cross, Megan; Hofmann, Andreas; Schröder, Rolf; Clemen, Christoph S; Eichinger, Ludwig
2016-01-01
p97 (VCP) is a homo-hexameric triple-A ATPase that exerts a plethora of cellular processes. Heterozygous missense mutations of p97 cause at least five human neurodegenerative disorders. However, the specific molecular consequences of p97 mutations are hitherto widely unknown. Our in silico structural models of human and Dictyostelium p97 showed that the disease-causing human R93C, R155H, and R155C as well as Dictyostelium R154C, E219K, R154C/E219K p97 mutations constitute variations in surface-exposed locations. In-gel ATPase activity measurements of p97 monomers and hexamers revealed significant mutation- and species-specific differences. While all human p97 mutations led to an increase in ATPase activity, no changes could be detected for the Dictyostelium R154C mutant, which is orthologous to human R155C. The E219K mutation led to an almost complete loss of activity, which was partially recuperated in the R154C/E219K double-mutant indicating p97 inter-domain communication. By means of co-immunoprecipitation experiments we identified an UBX-domain containing Dictyostelium protein as a novel p97 interaction partner. We categorized all UBX-domain containing Dictyostelium proteins and named the interaction partner UBXD9. Pull-down assays and surface plasmon resonance analyses of Dictyostelium UBXD9 or the human orthologue TUG/ASPL/UBXD9 demonstrated direct interactions with p97 as well as species-, mutation- and ATP-dependent differences in the binding affinities. Sucrose density gradient assays revealed that both human and Dictyostelium UBXD9 proteins very efficiently disassembled wild-type, but to a lesser extent mutant p97 hexamers into monomers. Our results are consistent with a scenario in which p97 point mutations lead to differences in enzymatic activities and molecular interactions, which in the long-term result in a late-onset and progressive multisystem disease. Copyright © 2016 The Authors. Published by Elsevier GmbH.. All rights reserved.
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Reduction of hydrogen desorption temperature of ball-milled MgH2 by NbF5 addition
International Nuclear Information System (INIS)
Recham, N.; Bhat, V.V.; Kandavel, M.; Aymard, L.; Tarascon, J.-M.; Rougier, A.
2008-01-01
Enhanced sorption properties of ball-milled MgH 2 are reported by adding NbF 5 . Among various catalyst amounts, 2 mol% of NbF 5 reveals to be the optimum concentration leading to significant reduction of the desorption temperature as well as faster kinetics of ball-milled MgH 2 . At 200 deg. C, temperature at which MgH 2 does not show any activity, MgH 2NbF 5 /2mol% composite desorbs 3.2 wt.% of H 2 in 50 mins. Interestingly, the addition of NbF 5 is also associated with an increase in the desorption pressure. At 300 deg. C, MgH 2NbF 5 /2mol% composite starts to desorb hydrogen at 600 mbar in comparison with 1 mbar for MgH 2 . Further improvements were successfully achieved by pre-grinding NbF 5 prior to ball-milling the catalyst with MgH 2 . Such pre-ground NbF 5 catalyzed MgH 2 composite desorbs 3 wt.% of H 2 at 150 deg. C. Improved properties are associated with smaller activation energies down to values close to the enthalpy of formation of MgH 2 . Finally, the mechanism at the origin of the enhancement is discussed in terms of catalyst stability, MgF 2 formation and electronic density localization
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Morphological and physiological investigations on mutants of Fusarium monoliforme IM
International Nuclear Information System (INIS)
Gancheva, V.
1996-01-01
High-producing mutants of Fusarium moniliforme IM are obtained as a result of gamma irradiation. The cultural characteristics of mutant strains 3284, 3211 and 76 following incubation of the producers for 14 days on potato-glucose agar are described. The colour of the aerial and substrate mycelium and the ability of the mutant strains to form conidiae and pigments are discussed in detail. The differences in the ability of mutants to assimilate different carbon and nitrogen sources are of specific importance for modelling nutrient media for submerged cultivation of F. moniliforme. 2 tabs., 2 figs. 7 refs
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Hypermutability of a UV-sensitive aphidicolin-resistant mutant of Chinese hamster fibroblasts
International Nuclear Information System (INIS)
Liu, P.K.; Chang, C.; Trosko, J.E.
1982-01-01
An ultraviolet light (UV)-sensitive thymidine auxotroph of Chinese hamster V79 cells that exhibits pleiotropic effects such as a high level of deoxycytidine triphosphate, slow growth, sensitivity to cytidine, and high frequencies of site-specific bromodeoxyuridine-dependent chromosomal aberrations was selected by its resistance to aphidicolin. The UV-induced mutability of this mutant and one of its revertants, which retains some of the phenotypes listed above, was studied in 3 mutation assay systems. The results showed that the mutant was hypermutable for ouabain and diphtheria-toxin-resistant mutations compared to wild-type V79 cells at the same UV dose or the same survival level. The mutant exhibits a delayed expression of maximal frequency of induced 6-thioguanine-resistant mutants. When maximal frequencies are compared at the same UV dose, the mutant also has higher mutation frequencies at the hypoxanthine-guanine phosphoribosyl transferase locus. The revertant was similar to the wild-type in UV sensitivity and mutability. (orig./AJ)
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Interaction of a non-peptide agonist with angiotensin II AT1 receptor mutants
DEFF Research Database (Denmark)
Costa-Neto, Claudio M; Miyakawa, Ayumi A; Pesquero, João B
2002-01-01
and inositol phosphate turnover assays in COS-7 cells transiently transfected with the wild-type and mutant forms of the receptor. Mutant receptors bore modifications in the extracellular region: T88H, Y92H, G1961, G196W, and D278E. Compound L-162,313 displaced [125I]-Sar1,Leu8-AngII from the mutants G196I...... and G196W with IC50 values similar to that of the wild-type. The affinity was, however, slightly affected by the D278E mutation and more significantly by the T88H and Y92H mutations. In inositol phosphate turnover assays, the ability of L-162,313 to trigger the activation cascade was compared...... with that of angiotensin II. These assays showed that the G196W mutant reached a relative maximum activation exceeding that of the wild-type receptor; the efficacy was slightly reduced in the G1961 mutant and further reduced in the T88H, Y92H, and D278E mutants. Our data suggest that residues of the extracellular domain...
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International Nuclear Information System (INIS)
Freeze, H.; Willies, L.; Hamilton, S.
1986-01-01
The lysosomal enzymes of Dictyostelium discoideum share highly immunogenic oligosaccharides which contain multiple Man-6-SO 4 residues. Two mutant strains which lack the shared antigenic determinant were analyzed in an attempt to identify the primary defect in each. [ 3 H]Man labelled N-linked oligosaccharides of secreted glycoproteins were released by Endo/PNGaseF digestion and analyzed. Both of the mutant strains produced smaller, less sulfated oligosaccharides compared to the wild-type, yet both still contained considerable amounts of Man-6-SO 4 . The size of the precursor lipid-linked oligosaccharide from the wild-type is consistent with a Glc 3 Man 9 GlcNAc 2 structure, while those from both of the mutants have an oligosaccharide the size of Man 5 GlcNAc 2 . The authors conclude that both of the mutants are defective in the biosynthesis of the precursor oligosaccharide. Both oligosaccharides from the mutants contain a tri-mannosyl core and are not glucosylated. Two of the five Man residues are released by a 1,2 specific α mannosidase. Based on the size and mannosidase digestions the authors conclude that 4/5 of the Man residues on the α1,6 branch of the β-linked Man residues are missing. Thus, these residues must be required to define the shared antigenic determinant
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Phenotypic and epistatic grouping of hypo- and hyper-rec mus mutants in Aspergillus.
Kafer, E; Chae, S K
1994-03-01
The mutants musK to musS of Aspergillus nidulans are sensitive to methyl-methanesulfonate (MMS) and several of them are meiotic-defective and alter mitotic recombination frequencies. All were found to be cross-sensitive to 4-nitro-quinoline-N-oxide (4-NQO) but unexpectedly none of them was hypersensitive to gamma-rays and few to UV light. Double mus; uvs mutants were constructed to test for interactions with uvs mutations of the four epistatic groups of Aspergillus, "UvsF", "UvsC", "UvsI", and "UvsB". All meiotic-defective mus mutations caused some lethal interactions, usually with uvsF. None of them showed epistasis with UvsF or UvsB group mutants and one, musO, may represent a new group. Three mus mutations that affect recombination were assigned to the UvsC group, namely musN and K, and also musL which is recombination-defective and closely resembles uvsC. While uvsC mutants are mutators and lack UV-mutagenesis, most mus mutants had no effects on mutation. Only musR, which appeared epistatic with uvsI, showed reduced UV-reversion frequencies similar to uvsI. The recombination-proficient mus mutants appeared to be epistatic with more than one group, but in several cases sensitivities were slight and overlaps insufficient to obtain corroborating results with MMS and 4-NQO.
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International Nuclear Information System (INIS)
Anton, D.N.
1979-01-01
Strain SB564 and its derivative DA78 are hypersensitive to the inhibitory action of the proteins coded for by genes hisF and hisH on cell division. Transduction of hisO1242, a regulatory mutation that elicits a very high level of expression of the histidine operon, into these strains resulted in the production of long filamentous cells carrying large balloons and in growth failure. Forty-one hisO1242 derivatives that escaped inhibition were isolated. These strains showed a large variety of alterations, many of which were related to the cell envelope. The more-frequent alterations included: changes in cell shape, increased sensitivity to one or more of several drugs (deoxycholate, cycloserine, penicillin, novobiocin, acridine orange), increased autolytic activity in alkaline buffer, anomalous fermentation of maltose on eosin--methylene blue plates, and temperature-conditional cell division. The alterations are produced, in some of the strains, by pleiotropic mutations in gene envB. Strains affected in divC, divD, and rodA loci have also been identified. Genetic analaysis has shown that several strains carry more than one envelope mutation. It is assumed that envelope mutations are positively selected because they somehow alleviate the particularly severe inhibition of cell division caused, in strains SB564 and DA78, by the excessive synthesis of hisF and hisH gene products
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Genetic Analysis and Molecular Mapping of a Novel Chlorophyll-Deficit Mutant Gene in Rice
Directory of Open Access Journals (Sweden)
Xiao-qun HUANG
2008-03-01
Full Text Available A rice etiolation mutant 824ys featured with chlorophyll deficiency was identified from a normal green rice variety 824B. It showed whole green-yellow plant from the seedling stage, reduced number of tillers and longer growth duration. The contents of chlorophyll, chlorophyll a, chlorophyll b and net photosynthetic rate in leaves of the mutant obviously decreased, as well as the number of spikelets per panicle, seed setting rate and 1000-grain weight compared with its wild-type parent. Genetic analyses on F1 and F2 generations of 824ys crossed with three normal green varieties showed that the chlorophyll-deficit mutant character was controlled by a pair of recessive nuclear gene. Genetic mapping of the mutant gene was conducted by using microsatellite markers and F2 mapping population of 495R/824ys, and the mutant gene of 824ys was mapped on the short arm of rice chromosome 3. The genetic distances from the target gene to the markers RM218, RM282 and RM6959 were 25.6 cM, 5.2 cM and 21.8 cM, respectively. It was considered to be a new chlorophyll-deficit mutant gene and tentatively named as chl11(t.
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McGrann, Graham R. D.; Steed, , Andrew; Burt, Christopher; Nicholson, Paul; Brown, James K. M.
2015-01-01
Lesion mimic mutants display spontaneous necrotic spots and chlorotic leaves as a result of mis-regulated cell death programmes. Typically these mutants have increased resistance to biotrophic pathogens but their response to facultative fungi that cause necrotrophic diseases is less well studied. The effect of altered cell death regulation on the development of disease caused by Ramularia collo-cygni, Fusarium culmorum and Oculimacula yallundae was explored using a collection of barley necrotic (nec) lesion mimic mutants. nec8 mutants displayed lower levels of all three diseases compared to nec9 mutants, which had increased R. collo-cygni but decreased F. culmorum disease symptoms. nec1 mutants reduced disease development caused by both R. collo-cygni and F. culmorum. The severity of the nec1-induced lesion mimic phenotype and F. culmorum symptom development was reduced by mutation of the negative cell death regulator MLO. The significant reduction in R. collo-cygni symptoms caused by nec1 was completely abolished in the presence of the mlo-5 allele and both symptoms and fungal biomass were greater than in the wild-type. These results indicate that physiological pathways involved in regulation of cell death interact with one another in their effects on different fungal pathogens. PMID:25873675
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Molecular Dynamics Driven Design of pH-Stabilized Mutants of MNEI, a Sweet Protein.
Directory of Open Access Journals (Sweden)
Serena Leone
Full Text Available MNEI is a single chain derivative of monellin, a plant protein that can interact with the human sweet taste receptor, being therefore perceived as sweet. This unusual physiological activity makes MNEI a potential template for the design of new sugar replacers for the food and beverage industry. Unfortunately, applications of MNEI have been so far limited by its intrinsic sensitivity to some pH and temperature conditions, which could occur in industrial processes. Changes in physical parameters can, in fact, lead to irreversible protein denaturation, as well as aggregation and precipitation. It has been previously shown that the correlation between pH and stability in MNEI derives from the presence of a single glutamic residue in a hydrophobic pocket of the protein. We have used molecular dynamics to study the consequences, at the atomic level, of the protonation state of such residue and have identified the network of intramolecular interactions responsible for MNEI stability at acidic pH. Based on this information, we have designed a pH-independent, stabilized mutant of MNEI and confirmed its increased stability by both molecular modeling and experimental techniques.
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Molecular Dynamics Driven Design of pH-Stabilized Mutants of MNEI, a Sweet Protein.
Leone, Serena; Picone, Delia
2016-01-01
MNEI is a single chain derivative of monellin, a plant protein that can interact with the human sweet taste receptor, being therefore perceived as sweet. This unusual physiological activity makes MNEI a potential template for the design of new sugar replacers for the food and beverage industry. Unfortunately, applications of MNEI have been so far limited by its intrinsic sensitivity to some pH and temperature conditions, which could occur in industrial processes. Changes in physical parameters can, in fact, lead to irreversible protein denaturation, as well as aggregation and precipitation. It has been previously shown that the correlation between pH and stability in MNEI derives from the presence of a single glutamic residue in a hydrophobic pocket of the protein. We have used molecular dynamics to study the consequences, at the atomic level, of the protonation state of such residue and have identified the network of intramolecular interactions responsible for MNEI stability at acidic pH. Based on this information, we have designed a pH-independent, stabilized mutant of MNEI and confirmed its increased stability by both molecular modeling and experimental techniques.
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Isolation of glutathione-deficient mutants of the yeast Saccharomyces cerevisiae
International Nuclear Information System (INIS)
Kistler, M.; Eckardt, F.; Summer, K.-H.
1986-01-01
Glutathione-deficient (gsh - ) mutants of the yeast Saccharomyces cerevisiae were isolated after UV treatment using MNNG as selective agent. For genetic and biochemical characterization 5 mutant strains were chosen which exhibited considerably decreased residual GSH contents varying from 2 to 6% of the wild-type levels. All 5 isolates showed a 2:2 segregation of the gsh - :GSH + phenotypes alluding to a monogenic recessive mutation. Complementation analysis indicates that all gsh - mutants belong to one complementation group. (Auth.)
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Directory of Open Access Journals (Sweden)
Zhu Li
Full Text Available The reversibility of thermal denaturation and catalytic efficiency of Bacillus licheniformis α-amylase were improved through site-directed mutagenesis. By using multiple sequence alignment and PoPMuSiC algorithm, Ser187 and Asn188, which located within a long loop in Domain B of Bacillus licheniformis α-amylase, were selected for mutation. In addition, Ala269, which is adjacent to Ser187 and Asn188, was also investigated. Seven mutants carrying the mutations S187D, N188T, N188S, A269K, A269K/S187D, S187D/N188T, and A269K/S187D/N188T were generated and characterized. The most thermostable mutant, A269K/S187D/N188T, exhibited a 9-fold improvement in half-life at 95°C and pH 5.5, compared with that of the wild-type enzyme. Mutant A269K/S187D/N188T also exhibited improved catalytic efficiency. The catalytic efficiency of mutant A269K/S187D/N188T reached 5.87×103±0.17 g·L-1·s-1 at pH 5.5, which is 1.84-fold larger than the corresponding value determined for the wild-type enzyme. Furthermore, the structure analysis showed that immobilization of the loop containing Ser187 and Asn188 plays a significant role in developing the properties of Bacillus licheniformis α-amylase.
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Li, Zhu; Duan, Xuguo; Chen, Sheng; Wu, Jing
2017-01-01
The reversibility of thermal denaturation and catalytic efficiency of Bacillus licheniformis α-amylase were improved through site-directed mutagenesis. By using multiple sequence alignment and PoPMuSiC algorithm, Ser187 and Asn188, which located within a long loop in Domain B of Bacillus licheniformis α-amylase, were selected for mutation. In addition, Ala269, which is adjacent to Ser187 and Asn188, was also investigated. Seven mutants carrying the mutations S187D, N188T, N188S, A269K, A269K/S187D, S187D/N188T, and A269K/S187D/N188T were generated and characterized. The most thermostable mutant, A269K/S187D/N188T, exhibited a 9-fold improvement in half-life at 95°C and pH 5.5, compared with that of the wild-type enzyme. Mutant A269K/S187D/N188T also exhibited improved catalytic efficiency. The catalytic efficiency of mutant A269K/S187D/N188T reached 5.87×103±0.17 g·L-1·s-1 at pH 5.5, which is 1.84-fold larger than the corresponding value determined for the wild-type enzyme. Furthermore, the structure analysis showed that immobilization of the loop containing Ser187 and Asn188 plays a significant role in developing the properties of Bacillus licheniformis α-amylase.
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Mutational analysis of the major soybean UreF paralogue involved in urease activation
Polacco, Joe C.; Hyten, David L.; Medeiros-Silva, Mônica; Sleper, David A.; Bilyeu, Kristin D.
2011-01-01
The soybean genome duplicated ∼14 and 45 million years ago and has many paralogous genes, including those in urease activation (emplacement of Ni and CO2 in the active site). Activation requires the UreD and UreF proteins, each encoded by two paralogues. UreG, a third essential activation protein, is encoded by the single-copy Eu3, and eu3 mutants lack activity of both urease isozymes. eu2 has the same urease-negative phenotype, consistent with Eu2 being a single-copy gene, possibly encoding a Ni carrier. Unexpectedly, two eu2 alleles co-segregated with missense mutations in the chromosome 2 UreF paralogue (Ch02UreF), suggesting lack of expression/function of Ch14UreF. However, Ch02UreF and Ch14UreF transcripts accumulate at the same level. Further, it had been shown that expression of the Ch14UreF ORF complemented a fungal ureF mutant. A third, nonsense (Q2*) allelic mutant, eu2-c, exhibited 5- to 10-fold more residual urease activity than missense eu2-a or eu2-b, though eu2-c should lack all Ch02UreF protein. It is hypothesized that low-level activation by Ch14UreF is ‘spoiled’ by the altered missense Ch02UreF proteins (‘epistatic dominant-negative’). In agreement with active ‘spoiling’ by eu2-b-encoded Ch02UreF (G31D), eu2-b/eu2-c heterozygotes had less than half the urease activity of eu2-c/eu2-c siblings. Ch02UreF (G31D) could spoil activation by Chr14UreF because of higher affinity for the activation complex, or because Ch02UreF (G31D) is more abundant than Ch14UreF. Here, the latter is favoured, consistent with a reported in-frame AUG in the 5' leader of Chr14UreF transcript. Translational inhibition could represent a form of ‘functional divergence’ of duplicated genes. PMID:21430294
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DEFF Research Database (Denmark)
Poulsen, Lars; Andersen, Mikael Rørdam; Lantz, Anna Eliasson
2012-01-01
exhibiting an oxalate overproducing phenotype were identified. The yield of oxalate was increased up to 158% compared to the wild type and the corresponding transcription factor was therefore entitled Oxalic Acid repression Factor, OafA. Detailed physiological characterization of one of the ΔoafA mutants......, compared to the wild type, showed that both strains produced substantial amounts of gluconic acid, but the mutant strain was more efficient in re-uptake of gluconic acid and converting it to oxalic acid, particularly at high pH (pH 5.0). Transcriptional profiles showed that 241 genes were differentially......Acid formation in Aspergillus niger is known to be subjected to tight regulation, and the acid production profiles are fine-tuned to respond to the ambient pH. Based on transcriptome data, putative trans-acting pH responding transcription factors were listed and through knock out studies, mutants...
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Polymorphisms in interleukins 17A and 17F genes and periodontitis: results from a meta-analysis.
da Silva, Felipe Rodolfo Pereira; Pessoa, Larissa Dos Santos; Vasconcelos, Any Carolina Cardoso Guimarães; de Aquino Lima, Weberson; Alves, Even Herlany Pereira; Vasconcelos, Daniel Fernando Pereira
2017-12-01
Polymorphisms in inflammatory genes such as interleukins 17A and 17F are associated with the risk of development of periodontitis, although the results remain contradictory. Hence, the aim of this study was perform a meta-analysis focusing on two polymorphisms (rs2275913 and rs763780) in interleukins 17A and 17F genes, respectively, in both chronic (CP) and aggressive periodontitis (AgP). A review in literature was performed in several databases for studies published before 25, September 2016. The meta-analysis was obtained through the review manager statistical software (version 5.2) with odds ratio (OR) calculation and funnel plot (P < 0.05) for heterogeneity, as well as the comprehensive meta-analysis software (version 3.3.070) for the assessment of publication bias. Seven articles with 1540 participants composed the results in which the mutant allele in the rs2275913 polymorphism did not present significant association with the risk of CP or AgP (OR 1.56, 95% CI 0.77, 3.15, P = 0.21; OR 1.12, 95% CI 0.05, 23.44, P = 0.94, respectively) nor was the mutant allele in rs763780 associated with the risk of CP (OR 1.19, 95% CI 0.80, 1.76, P = 0.39) or AgP (OR 1.07, 95% CI 0.63, 1.84, P = 0.79). No bias of publication was observed by Egger's and Begg's tests in any allelic evaluation. This meta-analysis showed a non-significant association between the polymorphisms rs2275913 and rs763780 in interleukins 17A and 17F genes and chronic and aggressive periodontitis in the allelic evaluation.
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Benomyl-resistant mutant strain of Trichoderma sp. with increased mycoparasitic activity.
Olejníková, P; Ondrusová, Z; Krystofová, S; Hudecová, D
2010-01-01
Application of UV radiation to the strain Trichoderma sp. T-bt (isolated from lignite) resulted in the T-brm mutant which was resistant to the systemic fungicide benomyl. The tub2 gene sequence in the T-brm mutant differed from the parent as well as the collection strain (replacing tyrosine with histidine in the TUB2 protein). Under in vitro conditions this mutant exhibited a higher mycoparasitic activity toward phytopathogenic fungi.
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Hama, S; Kimura, G
1980-01-01
Eleven temperature-sensitive mutants of adenovirus type 12, capable of forming plaques in human cells at 33 C but not at 39.5 C, were isolated from a stock of a wild-type strain after treatment with either nitrous acid or hydroxylamine. Complementation tests in doubly infected human cells permitted a tentative assignment of eight of these mutants to six complementation groups. Temperature-shift experiments revealed that one mutant is affected early and most of the other mutants are affected late. Only the early mutant, H12ts505, was temperature sensitive in viral DNA replication. Infectious virions of all the mutants except H12ts505 and two of the late mutants produced at 33 C, appeared to be more heat labile than those of the wild type. Only H12ts505 was temperature sensitive for the establishment of transformation of rat 3Y1 cells. One of the late mutants (H12ts504) had an increased transforming ability at the permissive temperature. Results of temperature-shift transformation experiments suggest that a viral function affected in H12ts505 is required for "initiation" of transformation. Some of the growth properties of H12ts505-transformed cells were also temperature dependent, suggesting that a functional expression of a gene mutation in H12ts505 is required to maintain at least some aspects of the transformed state.
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2000-01-01
H1 is one of the two large detectors installed at HERA, the first electron-proton accelerator, located at DESY in Hamburg. The H1 collaboration regroups physicists from 32institutes of 11countries all over the world.
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DEFF Research Database (Denmark)
Mohey, Rajesh; Jørgensen, Anne Louise; Møller, Bjarne K
2005-01-01
and incorporation of resistant forms in the long-lived CD4+ T cellular DNA compartment is not clear. Objective To investigate the relationship between lamivudine associated mutant-type 184V and the wild-type 184M proviral forms in the circulating CD4+ T cells of patients and low-level viremia. Study design Cross-sectional......Background Highly active anti-retroviral therapy (HAART) effectively reduces HIV replication but does not completely hinder it. Sub-optimal therapy leads to HIV resistance to the drugs administered. However, the role of low-level viremia (viral-load less than 1000 copies/ml) on mutation genesis...... study of 50 patients on long-term HAART, with a viremia of less than 1000 copies/ml. Patients were stratified into three groups; on lamivudine, group I (viral load
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DEFF Research Database (Denmark)
Willumsen, B M; Vass, W C; Velu, T J
1991-01-01
We have previously used a series of insertion-deletion mutants of the mutationally activated v-rasH gene to identify several regions of the encoded protein that are dispensable for cellular transformation (B. M. Willumsen, A. G. Papageorge, H.-F. Kung, E. Bekesi, T. Robins, M. Johnsen, W. C. Vass...... in their v-rasH forms. We conclude that a region including amino acids 102 and 103 encodes a function that is more critical to c-rasH than to v-rasH. Guanine nucleotide exchange is one function that is compatible with such a phenotype......., and D. R. Lowy, Mol. Cell. Biol. 6:2646-2654, 1986). To determine if some of these amino acids are more important for the biological activity of c-rasH, we have now tested many of the same insertion-deletion mutants in the c-rasH form for their ability to transform NIH 3T3 cells. Since the transforming...
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Energy Technology Data Exchange (ETDEWEB)
Buragohain, Amlan [Department of Chemistry, Indian Institute of Technology Guwahati, 781039 Assam (India); Couck, Sarah [Department of Chemical Engineering, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels (Belgium); Van Der Voort, Pascal [Department of Inorganic and Physical Chemistry, Ghent University, COMOC – Center for Ordered Materials, Organometallics and Catalysis, Krijgslaan 281-S3, 9000 Ghent (Belgium); Denayer, Joeri F.M. [Department of Chemical Engineering, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels (Belgium); Biswas, Shyam, E-mail: sbiswas@iitg.ernet.in [Department of Chemistry, Indian Institute of Technology Guwahati, 781039 Assam (India)
2016-06-15
Four existing and three new functionalized chromium terephthalates having MIL-101 topology and denoted as Cr-MIL-101-X (existing ones with X=–F, 1-F; –Cl, 2-Cl; –Br, 3-Br; –CH{sub 3}, 4-CH{sub 3}; new ones with X=–C{sub 6}H{sub 4}, 5-C{sub 6}H{sub 4}; –F{sub 2}, 6-F{sub 2}, –(CH{sub 3}){sub 2}, 7-(CH{sub 3}){sub 2}) were synthesized under hydrothermal conditions. All the materials except 5-C{sub 6}H{sub 4} could be prepared by a general synthetic route, in which the mixtures of CrO{sub 3}, H{sub 2}BDC-X (BDC=1,4-benzenedicarboxylate) linkers, conc. HCl and water with a molar ratio of 1:1:3.9:222.2 were reacted at 180 °C for 144 h. Compared to the 144 h of synthesis time, three of the compounds, namely 1-Cl, 2-Br and 5-C{sub 6}H{sub 4}, could be prepared in much shorter reaction times (12–18 h at 180–210 °C). The materials possess high thermal stability up to 270–300 °C in an air atmosphere. The activated compounds exhibit significant porosity (S{sub BET} range: 1273–2135 m{sup 2} g{sup −1}). At 0 °C and 1 bar, the CO{sub 2} adsorption capacities of the compounds fall in the 1.7–2.9 mmol g{sup −1} range. Compounds 1-F and 6-F{sub 2} showed enhanced CO{sub 2} uptake values compared to parent Cr-MIL-101. The benzene adsorption capacities of the compounds lie in the range of 66.2–139.5 molecules per unit cell at 50 °C and p/p{sub 0}=0.35. The increased benzene uptake value of 1-F compared to un-functionalized Cr-MIL-101 and 4-CH{sub 3} suggests that the fluorination has induced more hydrophobicity in Cr-MIL-101 as compared to the methylation. - Graphical abstract: Benzene adsorption by seven functionalized Cr-MIL-101-X metal-organic framework (MOF) materials Display Omitted - Highlights: • Seven functionalized Cr-MIL-101-X materials were synthesized solvothermally. • All Cr-MIL-101-X materials exhibited high thermal stability up to 270–300 °C in air. • All Cr-MIL-101-X compounds displayed considerable porosity towards N{sub 2
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Mechanical properties of TIG and EB weld joints of F82H
Energy Technology Data Exchange (ETDEWEB)
Hirose, Takanori, E-mail: hirose.takanori@jaea.go.jp; Sakasegawa, Hideo; Nakajima, Motoki; Tanigawa, Hiroyasu
2015-10-15
Highlights: • Narrow groove TIG minimized volume of F82H weld. • Mechanical properties of TIG and EB welds of F82H have been characterized. • Post weld heat treatment successfully moderate the toughness of weld metal without softening the base metal. - Abstract: This work investigates mechanical properties of weld joints of a reduced activation ferritic/martensitic steel, F82H and effects of post weld heat treatment on the welds. Vickers hardness, tensile and Charpy impact tests were conducted on F82H weld joints prepared using tungsten-inert-gas and electron beam after various heat treatments. Although narrow groove tungsten-inert-gas welding reduced volume of weld bead, significant embrittlement was observed in a heat affected zone transformed due to heat input. Post weld heat treatment above 993 K successfully moderated the brittle transformed region. The hardness of the brittle region strongly depends on the heat treatment temperature. Meanwhile, strength of base metal was slightly reduced by the treatment at temperature ranging from 993 to 1053 K. Moreover, softening due to double welding was observed only in the weld metal, but negligible in base metal.
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Ligase-deficient yeast cells exhibit defective DNA rejoining and enhanced gamma ray sensitivity
International Nuclear Information System (INIS)
Moore, C.W.
1982-01-01
Yeast cells deficient in DNA ligase were also deficient in their capacity to rejoin single-strand scissions in prelabeled nuclear DNA. After high-dose-rate gamma irradiation (10 and 25 krads), cdc9-9 mutant cells failed to rejoin single-strand scissions at the restrictive temperature of 37 0 C. In contrast, parental (CDC9) cells (incubated with mutant cells both during and after irradiation) exhibited rapid medium-independent DNA rejoining after 10 min of post-irradiation incubation and slower rates of rejoining after longer incubation. Parental cells were also more resistant than mutant cells to killing by gamma irradiation. Approximately 2.5 +- 0.07 and 5.7 +- 0.6 single-strand breaks per 10 8 daltons were detected in DNAs from either CDC9 or cdc9-9 cells converted to spheroplasts immediately after 10 and 25 krads of irradiation, respectively. At the permissive temperature of 23 0 C, the cdc9-9 cells contained 2 to 3 times the number of DNA single-strand breaks as parental cells after 10 min to 4 h of incubation after 10 krads of irradiation, and two- to eightfold more breaks after 10 min to 2.5 h of incubation after 25 krads of irradiation. Rejoining of single-strand scissions was faster in medium. After only 10 min in buffered growth medium after 10 krads of irradiation, the number of DNA single-strand breaks was reduced to 0.32 +- 0.3 (at 23 0 C) or 0.21 +- 0.05 (at 37 0 C) per 10 8 daltons in parental cells, but remained at 2.1 +- 0.06 (at 23 0 C) or 2.3 +- 0.07 (at 37 0 C) per 10 8 daltons in mutant cells. After 10 or 25 krads of irradiation plus 1 h of incubation in medium at 37 0 C, only DNA from CDC9 cells was rejoined to the size of DNA from unirradiated cells, whereas at 23 0 C, DNAs in both strains were completely rejoined
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Cytogenetic characteristics of soft wheat mutants under x-irradiation
International Nuclear Information System (INIS)
Shakaryan, Zh.O.; Avakyan, V.A.; Amirbekyan, V.A.
1981-01-01
Radiosensitivity of induced mutants of soft wheat is studied by criteria of frequency and character of changes in 1 and 2 divisions of meiosis. Two constant induced mutant forms of soft wheat were investigated. Mutant lines of squareheads with red ear (re) and erectoids 37/1 were obtained by X-ray irradiating hydride seeds F 1 of hybride combination of Alty-Agach Awnless 1. Seeds of mutants and initial kinds were exposed to X-rays at a dose of 10 kR. A conclusion may be drawn on the basis of studying the meiosis process in mutants and initial kinds of soft wheat on X-ray radiation that the mutants are more radiosensitive. This testifies to that that the induced mutants of soft wheat represent new genotypes in comparison with the initial kinds and differ from the latter not only in morphological characters but in the reaction norm with respect to external medium factors, i.e. the limit of possible changeability of the genotype has been extended [ru
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DNA binding properties of dioxin receptors in wild-type and mutant mouse hepatoma cells
International Nuclear Information System (INIS)
Cuthill, S.; Poellinger, L.
1988-01-01
The current model of action of 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin) entails stimulation of target gene transcription via the formation of dioxin-receptor complexes and subsequent accumulation of the complexes within the cell nucleus. Here, the authors have analyzed the DNA binding properties of the dioxin receptor in wild-type mouse hepatoma (Hepa 1c1c7) cells and a class of nonresponsive mutant cells which fail to accumulate dioxin-receptor complexes within the nucleus in vivo. In vitro, both the wild-type and mutant [ 3 H]dioxin-receptor complexes exhibited low affinity for DNA-cellulose (5-8% and around 4% retention, respectively) in the absence of prior biochemical manipulations. However, following chromatography on heparin-Sepharose, the wild-type but not the mutant dioxin receptor was transformed to a species with an increased affinity for DNA (40-50% retention on DNA-cellulose). The gross molecular structure of the mutant, non DNA binding dioxin receptor did not appear to be altered as compared to that of the wild-type receptor. These results imply that the primary deficiency in the mutant dioxin receptor form may reside at the DNA binding level and that, in analogy to steroid hormone receptors, DNA binding of the receptor may be an essential step in the regulation of target gene transcription by dioxin
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Rate and Regulation of Copper Transport by Human Copper Transporter 1 (hCTR1)*
Maryon, Edward B.; Molloy, Shannon A.; Ivy, Kristin; Yu, Huijun; Kaplan, Jack H.
2013-01-01
Human copper transporter 1 (hCTR1) is a homotrimer of a 190-amino acid monomer having three transmembrane domains believed to form a pore for copper permeation through the plasma membrane. The hCTR1-mediated copper transport mechanism is not well understood, nor has any measurement been made of the rate at which copper ions are transported by hCTR1. In this study, we estimated the rate of copper transport by the hCTR1 trimer in cultured cells using 64Cu uptake assays and quantification of plasma membrane hCTR1. For endogenous hCTR1, we estimated a turnover number of about 10 ions/trimer/s. When overexpressed in HEK293 cells, a second transmembrane domain mutant of hCTR1 (H139R) had a 3-fold higher Km value and a 4-fold higher turnover number than WT. Truncations of the intracellular C-terminal tail and an AAA substitution of the putative metal-binding HCH C-terminal tripeptide (thought to be required for transport) also exhibited elevated transport rates and Km values when compared with WT hCTR1. Unlike WT hCTR1, H139R and the C-terminal mutants did not undergo regulatory endocytosis in elevated copper. hCTR1 mutants combining methionine substitutions that block transport (M150L,M154L) on the extracellular side of the pore and the high transport H139R or AAA intracellular side mutations exhibited the blocked transport of M150L,M154L, confirming that Cu+ first interacts with the methionines during permeation. Our results show that hCTR1 elements on the intracellular side of the hCTR1 pore, including the carboxyl tail, are not essential for permeation, but serve to regulate the rate of copper entry. PMID:23658018
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Cai, Xun-Chao; Xi, Huan; Liang, Li; Liu, Jia-Dong; Liu, Chang-Hong; Xue, Ya-Rong; Yu, Xiang-Yang
2017-01-01
Rifampicin resistance (Rif r ) mutations in the RNA polymerase β subunit ( rpoB ) gene exhibit pleiotropic phenotypes as a result of their effects on the transcription machinery in prokaryotes. However, the differences in the effects of the mutations on the physiology and metabolism of the bacteria remain unknown. In this study, we isolated seven Rif r mutations in rpoB , including six single point mutations (H485Y, H485C, H485D, H485R, Q472R, and S490L) and one double point mutation (S490L/S617F) from vegetative cells of an endophytic strain, Bacillus velezensis CC09. Compared to the wild-type (WT) strain (CC09), the H485R and H485D mutants exhibited a higher degree of inhibition of Aspergillus niger spore germination, while the H485Y, S490L, Q472R, and S490L/S617F mutants exhibited a lower degree of inhibition due to their lower production of the antibiotic iturin A. These mutants all exhibited defective phenotypes in terms of pellicle formation, sporulation, and swarming motility. A hierarchical clustering analysis of the observed phenotypes indicated that the four mutations involving amino acid substitutions at H485 in RpoB belonged to the same cluster. In contrast, the S490L and Q472R mutations, as well as the WT strain, were in another cluster, indicating a functional connection between the mutations in B. velezensis and phenotypic changes. Our data suggest that Rif r mutations cannot only be used to study transcriptional regulation mechanisms, but can also serve as a tool to increase the production of bioactive metabolites in B. velezensis .
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Human GLTP and mutant forms of ACD11 suppress cell death in the Arabidopsis acd11 mutant
DEFF Research Database (Denmark)
Petersen, Nikolaj H T; McKinney, Lea V; Pike, Helen
2008-01-01
The Arabidopsis acd11 mutant exhibits runaway, programmed cell death due to the loss of a putative sphingosine transfer protein (ACD11) with homology to mammalian GLTP. We demonstrate that transgenic expression in Arabidopsis thaliana of human GLTP partially suppressed the phenotype of the acd11...
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A study of analysis PB1-F2 protein of Influenza Viruses A/H1N1pdm09, A/ H3N2, and A/H5N1
Directory of Open Access Journals (Sweden)
Hana Apsari Pawestri
2016-07-01
Full Text Available Abstrak Tujuan. Protein PB1-F2 (polymerase basic 1-frame 2 adalah protein terbaru yang ditemukan pada virus Influenza dan telah terbukti berperan dalam induksi kematian sel dan patogenitas. Tujuan dari tulisan ini adalah untuk menganalisis protein PB1-F2 pada virus Influenza A/H5N1 dan A/H1N1pdm09. Metode. Kami melakukan pencarian data yang relevan yaitu sekuens gen virus Influenza A/H5N1 dan A/H1N1pdm09 dari Gen Bank National Center for Biotechnology Information (NCBI selama tahun 1997-2015. Data yang digunakan adalah data sekuens nukleotida gen PB1 (polymerase basic1 virus influenza A/H5N1 dan A/H1N1pdm09. Kemudian dilakukan analisis alignment untuk mengetahui variasi protein dan mutasi yang berhubungan dengan patogenitas dan virulensi. Hasil. Kami melakukan penelitian terhadap sekuens PB1-F2 sebanyak 3262 influenza A/H5N1 dan 2472 Influenza A/H1N1pdm09. Hasil analisis menunjukkan bahwa semua sekuens A/H5N1 memiliki panjang yang penuh sebanyak 90 asam amino, kecuali influenza pandemi 2009 hanya memiliki panjang 87 asam amino. Kemudian, ditemukan mutasi yang berhubungan dengan virulensi yang ditunjukan dengan perubahan asam amino Asparagin (N menjadi Serin (S. Mutasi tersebut terjadi pada Influenza A/H5N1 sebanyak 8.5% dan Influenza A/H1N1pdm09 sebanyak 0.5%. Kesimpulan. Ditemukan beberapa variasi panjang asam amino dan mutasi penting pada sekuens PB1-F2 dari subtipe yang berbeda yaitu influenza A/H5N1 dan A/H1N1pdm09 yang mengindikasikan seleksi spesifik karena introduksi dan adaptasi terhadap inang yang berbeda. Diperlukan penelitian lanjutan untuk lebih memahami variasi dan kontribusi protein PB1-F2 tersebut terhadap virulensi dan patogenitas virus Influenza. Kata kunci : Patogenesis, Virus Influenza, Protein PB1-F2 Abstract Aim. Influenza virus PB1-F2 (polymerase basic 1-frame 2 protein is a novel protein previously shown to be involved in cell death induction and pathogenesis. Here we analysis the PB1-F2 protein of Influenza virus A/H
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A study of analysis PB1-F2 protein of Influenza Viruses A/H1N1pdm09, A/ H3N2, and A/H5N1
Directory of Open Access Journals (Sweden)
Hana Apsari Pawestri
2016-07-01
Full Text Available Abstrak Tujuan. Protein PB1-F2 (polymerase basic 1-frame 2 adalah protein terbaru yang ditemukan pada virus Influenza dan telah terbukti berperan dalam induksi kematian sel dan patogenitas. Tujuan dari tulisan ini adalah untuk menganalisis protein PB1-F2 pada virus Influenza A/H5N1 dan A/H1N1pdm09. Metode. Kami melakukan pencarian data yang relevan yaitu sekuens gen virus Influenza A/H5N1 dan A/H1N1pdm09 dari Gen Bank National Center for Biotechnology Information (NCBI selama tahun 1997-2015. Data yang digunakan adalah data sekuens nukleotida gen PB1 (polymerase basic1 virus influenza A/H5N1 dan A/H1N1pdm09. Kemudian dilakukan analisis alignment untuk mengetahui variasi protein dan mutasi yang berhubungan dengan patogenitas dan virulensi. Hasil. Kami melakukan penelitian terhadap sekuens PB1-F2 sebanyak 3262 influenza A/H5N1 dan 2472 Influenza A/H1N1pdm09. Hasil analisis menunjukkan bahwa semua sekuens A/H5N1 memiliki panjang yang penuh sebanyak 90 asam amino, kecuali influenza pandemi 2009 hanya memiliki panjang 87 asam amino. Kemudian, ditemukan mutasi yang berhubungan dengan virulensi yang ditunjukan dengan perubahan asam amino Asparagin (N menjadi Serin (S. Mutasi tersebut terjadi pada Influenza A/H5N1 sebanyak 8.5% dan Influenza A/H1N1pdm09 sebanyak 0.5%. Kesimpulan. Ditemukan beberapa variasi panjang asam amino dan mutasi penting pada sekuens PB1-F2 dari subtipe yang berbeda yaitu influenza A/H5N1 dan A/H1N1pdm09 yang mengindikasikan seleksi spesifik karena introduksi dan adaptasi terhadap inang yang berbeda. Diperlukan penelitian lanjutan untuk lebih memahami variasi dan kontribusi protein PB1-F2 tersebut terhadap virulensi dan patogenitas virus Influenza. Kata kunci : Patogenesis, Virus Influenza, Protein PB1-F2 Abstract Aim. Influenza virus PB1-F2 (polymerase basic 1-frame 2 protein is a novel protein previously shown to be involved in cell death induction and pathogenesis. Here we analysis the PB1-F2 protein of Influenza virus A/H
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46 CFR 184.702 - Pollution prevention equipment and procedures.
2010-10-01
... 46 Shipping 7 2010-10-01 2010-10-01 false Pollution prevention equipment and procedures. 184.702 Section 184.702 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS... Pollution prevention equipment and procedures. A vessel must comply with the applicable design, equipment...
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International Nuclear Information System (INIS)
Zelitch, I.
1990-01-01
The increase in net photosynthesis in M 4 progeny of an O 2 -resistant tobacco (Nicotiana tabacum) mutant relative to wild-type plants at 21 and 42% O 2 has been confirmed and further investigated. Self-pollination of an M 3 mutant produced M 4 progeny segregating high catalase phenotypes (average 40% greater than wild type) at a frequency of about 60%. The high catalase phenotype cosegregated precisely with O 2 -resistant photosynthesis. About 25% of the F 1 progeny of reciprocal crosses between the same M 3 mutant and wild type had high catalase activity, whether the mutant was used as the maternal or paternal parent, indicating nuclear inheritance. In high-catalase mutants the activity of NADH-hydroxypyruvate reductase, another peroxisomal enzyme, was the same as wild type. The mutants released 15% less photorespiratory CO 2 as a percent of net photosynthesis in CO 2 -free 21% O 2 and 36% less in CO 2 -free 42% O 2 compared with wild type. The mutant leaf tissue also released less 14 CO 2 per [1- 14 C]glycolate metabolized than wild type in normal air, consistent with less photorespiration in the mutant. The O 2 -resistant photosynthesis appears to be caused by a decrease in photorespiration especially under conditions of high O 2 where the stoichiometry of CO 2 release per glycolate metabolized is expected to be enhanced. The higher catalase activity in the mutant may decrease the nonenzymatic peroxidation of keto-acids such as hydroxypyruvate and glyoxylate by photorespiratory H 2 O 2
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International Nuclear Information System (INIS)
Masur, Shakuntala; Madhusudan, K.
2009-01-01
Sesame induced mutants were isolated from gamma irradiation of a local variety DS-1. The promising mutants for high yield and bold seed were utilized for assessing lignan profiles mainly sesamin, sesamolin and gamma tocopherol.mutant-1022 exhibited 146 per cent improvement over the parent, Since, the sesame possess numerous health benefits, the value added confectionary products were prepared. The studies revealed that, the biscuits prepared by using 100 per cent sesame flour exhibited very good spreading quality and baking strength. Organoleptic score indicated that 80% of the evaluators accepted biscuits prepared from pure sesame flour. (author)
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Wang, Yan; Liu, Fei; Wang, Liuqing; Wang, Qi; Selvaraj, Jonathan Nimal; Zhao, Yueju; Wang, Yun; Xing, Fuguo; Liu, Yang
2018-05-02
In Aspergillus and Penicillium species, an essential pH-response transcription factor pacC is involved in growth, pathogenicity, and toxigenicity. To investigate the connection between ochratoxin A (OTA) biosynthesis and ambient pH, the AopacC in Aspergillus ochraceus was functionally characterized using a loss-of-function mutant. The mycelium growth was inhibited under pH 4.5 and 10.0, while the sporulation increased under alkaline condition. A reduction of mycelium growth and an elevation of sporulation was observed in Δ AopacC mutant. Compared to neutral condition, OTA contents were respectively reduced by 71.6 and 79.8% under acidic and alkaline conditions. The expression of AopacC increased with the elevated pH, and deleting AopacC dramatically decreased OTA production and biosynthetic genes Aopks expression. Additionally, the Δ AopacC mutant exhibited attenuated infection ability toward pear fruits. These results suggest that AopacC is an alkaline-induced regulator responsible for growth and OTA biosynthesis in A. ochraceus and this regulatory mechanism might be pH-dependent.
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International Nuclear Information System (INIS)
Boshra, S.A.; Azer, S.A.
2004-01-01
some promising mutants of rice variety Giza 180 induced by using gamma rays were tested for susceptibility to infestation by angoumois grain moth, sitotroga cerealella in relation with grain physical characters. number of F1 progeny and percentage of adult emergence showed that mutants 7/1, 4/5 and 1/7 were the most susceptible o angoumois grain moth, while mutants 13/1,16/1 and 15/6 b were least susceptible . there was positive correlation between number of F1 adult progeny and grain weight loss. the development period of the insect ranged from 33.7 days on 1/7 to 42.9 days on 15/6 b and the adults that emerged from mutants 1/7,4/5,7/1,8/1,15/1 and 11/1 lived longer than those of the other ones. Grains of the mutants with thick and pubescent or intermediate hulls were the least susceptible to attack by this pest. the rice mutants 13/1,16/1,15/6 b and their parental cultivar Giza 180 had the highest grain yield
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Photo dynamics of BLUF domain mutant H44R of AppA from Rhodobacter sphaeroides
International Nuclear Information System (INIS)
Zirak, P.; Penzkofer, A.; Hegemann, P.; Mathes, T.
2007-01-01
The photo-cycle dynamics of the H44R mutant of the BLUF domain of the transcriptional anti-repressor protein AppA (AppA-H44R) from the non-sulfur anoxyphototropic purple bacterium Rhodobacter sphaeroides is studied in order to gain information on the involvement of His44 in the photo-cyclic mechanism of the AppA BLUF domain and to add information to the involved processes. The amino acid residue histidine at position 44 is replaced by arginine. A 12 nm red-shifted signalling state is formed upon blue-light excitation, while in wild-type AppA (AppA-wt) the red-shift is 16 nm. The recovery to the receptor dark state is approximately a factor of 2.5 faster (τ rec ∼ 6.5 min) than the recovery of the wild-type counterpart. Extended light exposure of the mutant causes photo-degradation of flavin (mainly free flavin conversion to lumichrome and re-equilibration between free and non-covalently bound flavin) and protein aggregation (showing up as light scattering). No photo-degradation was observed for AppA-wt. The quantum efficiency of signalling-state formation determined by intensity dependent absorption measurements is found to be φ s ∼ 0.3 (for AppA-wt: φ s ∼ 0.24). A two-component single-exponential fluorescence relaxation was observed, which is interpreted as fast fluorescence quenching to an equilibrium value by photo-induced electron transfer followed by slower fluorescence decay due to charge recombination. Based on the experimental findings, an extended photo-cycle model for BLUF domains is proposed
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Photo dynamics of BLUF domain mutant H44R of AppA from Rhodobacter sphaeroides
Zirak, P.; Penzkofer, A.; Hegemann, P.; Mathes, T.
2007-05-01
The photo-cycle dynamics of the H44R mutant of the BLUF domain of the transcriptional anti-repressor protein AppA (AppA-H44R) from the non-sulfur anoxyphototropic purple bacterium Rhodobacter sphaeroides is studied in order to gain information on the involvement of His44 in the photo-cyclic mechanism of the AppA BLUF domain and to add information to the involved processes. The amino acid residue histidine at position 44 is replaced by arginine. A 12 nm red-shifted signalling state is formed upon blue-light excitation, while in wild-type AppA (AppA-wt) the red-shift is 16 nm. The recovery to the receptor dark state is approximately a factor of 2.5 faster ( τrec ≈ 6.5 min) than the recovery of the wild-type counterpart. Extended light exposure of the mutant causes photo-degradation of flavin (mainly free flavin conversion to lumichrome and re-equilibration between free and non-covalently bound flavin) and protein aggregation (showing up as light scattering). No photo-degradation was observed for AppA-wt. The quantum efficiency of signalling-state formation determined by intensity dependent absorption measurements is found to be ϕs ≈ 0.3 (for AppA-wt: ϕs ≈ 0.24). A two-component single-exponential fluorescence relaxation was observed, which is interpreted as fast fluorescence quenching to an equilibrium value by photo-induced electron transfer followed by slower fluorescence decay due to charge recombination. Based on the experimental findings, an extended photo-cycle model for BLUF domains is proposed.
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International Nuclear Information System (INIS)
Engvild, K.C.; Rasmussen, K.
2005-01-01
Barley mutants without root hairs or with short or reduced root hairs were isolated among M 2 seeds of 'Lux' barley (Hordeum vulgare L.) after acidified sodium azide mutagenesis. Root hair mutants are investigated intensively in Arabidopsis where about 40 genes are known. A few root hair mutants are known in maize, rice, barley and tomato. Many plants without root hairs grow quite well with good plant nutrition, and mutants have been used for investigations of uptake of strongly bound nutrients like phosphorus, iron, zinc and silicon. Seed of 'Lux' barley (Sejet Plant Breeding, Denmark) were soaked overnight, and then treated with 1.5-millimolarsodium azide in 0.1 molar sodium phosphate buffer, pH 3, for 2.5 hours according to the IAEA Manual on Mutation Breeding (2nd Ed.). After rinsing in tap water and air-drying, the M 2 seeds were sown in the field the same day. Spikes, 4-6 per M 1 plant, were harvested. The mutation frequency was similar to that obtained with other barley cultivars from which low-phytate mutants were isolated [5]. Seeds were germinated on black filter paper in tap water for 3 or 4 days before scoring for root hair mutants
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Reversibility of U → F processes in X or UV irradiated KCl: H- systems
International Nuclear Information System (INIS)
Bressiani, Ana Helena A.
1979-01-01
Potassium Chloride crystals, pure, additively colored or hydrogenated have been exposed to X and UV radiations for correlation studies' of formation processes of defects. In hydrogenated samples, these irradiations are responsible-for a direct U→F centers conversion with the simultaneous creation of H 2 molecules besides the intrinsic formation of F/hole centers. After prolonged irradiation, F aggregate centers react with H 2 molecules forming U center pairs. As the two U centers have independent electronic transitions they add up, increasing the maximum height of the U band after F→U reversed reation. The saturation levels of defects produced by these two types of radiation are different because UV light also excites F centers while producing them. This effect is responsible by the F center aggregation and the inversion F→U decreasing the U→F efficiency. Crystals with F and U centers under F light also show F→U inversion but with higher efficiency than when under UV light. This F→U process gives back 90% of the initial U center concentration. This same F→U process was observed but at smaller rate of formation if samples containing U, F and F aggregates are left in the dark for prolonged periods of time. The same reactions take place namely H 2 molecules annihilation F aggregates. (author)
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Energy Technology Data Exchange (ETDEWEB)
Brunet, A; Geller, R [Commissariat a l' Energie Atomique, Saclay (France).Centre d' Etudes Nucleaires
1960-07-01
In the first part of this paper, we study the single Langmuir probe and the double-probe method. In the second part, we describe the probe-technique in connection with R.F. plasmoids. (author) [French] Dans la premiere partie de cet article, nous rappelons les principes d'utilisation de la sonde simple de Langmuir ainsi que la methode de la sonde double. La seconde partie est consacree a la pratique des sondes dans l'etude des plasmoides H.F. (auteur)
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SRP engineering and design history, Vol III, 200 F and H Areas
International Nuclear Information System (INIS)
Banick, C.J.
2000-01-01
This volume combines the record of events relating to the development of design for both the 200-F and H Areas. Chronologically, the definition of plant facilities was first established for the 200-F Area. The second area, 200-H, was projected initially to be a supplementary plutonium separations facility. This history explains the differences in character and capacity of the manufacturing facilities in both areas as production requirements and experience with separations processes advanced
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SRP engineering and design history, Vol III, 200 F and H Areas
Energy Technology Data Exchange (ETDEWEB)
Banick, C.J.
2000-04-17
This volume combines the record of events relating to the development of design for both the 200-F and H Areas. Chronologically, the definition of plant facilities was first established for the 200-F Area. The second area, 200-H, was projected initially to be a supplementary plutonium separations facility. This history explains the differences in character and capacity of the manufacturing facilities in both areas as production requirements and experience with separations processes advanced.
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Biswal, Jitendra K; Das, Biswajit; Sharma, Gaurav K; Khulape, Sagar A; Pattnaik, Bramhadev
2016-04-01
Foot-and-mouth disease virus (FMDV) particles lose infectivity due to their dissociation into pentamers at pH value below 6.5. After the uptake of FMDV by receptor-mediated endocytosis, the acid-dependent dissociation process is required for the release of FMDV genome inside endosomes. Nevertheless, dissociation of FMDV particles in mildly acidic conditions renders the inactivated FMD vaccine less effective. To improve the acid stability of inactivated FMD vaccine during the manufacturing process, a serotype A IND 40/2000 (in-use vaccine strain) mutant with increased resistance to acid inactivation was generated through reverse genetics approach. Based upon the earlier reports, the crucial amino acid residue, H142 of VP3 capsid protein was substituted separately to various amino acid residues Arg (R), Phe (F), Ala (A), and Asp (D) on the full-genome length cDNA clone. While the H142 → R or H142 → F or H142 → A substitutions resulted in non-infectious FMDV, H142 → D mutation on VP3 protein (H3142D) resulted in the generation of mutant virus with enhanced resistance to acid-induced inactivation. In addition, H3142D substitution did not alter the replication ability and antigenicity of mutant as compared to the parental virus. However, the virus competition experiments revealed that the H3142D substitution conferred a loss of fitness for the mutant virus. Results from this study demonstrate that the H3142D substitution is the molecular determinant of acid-resistant phenotype in FMDV serotype A.
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Selection of high hectolitre weight mutants of winter wheat
International Nuclear Information System (INIS)
Crowley, C.; Jones, P.
1989-01-01
Grain quality in wheat includes hectolitre weight (HLW) besides protein content and thousand-grain weight (TGW). The British winter wheat variety ''Guardian'' has a very high yield potential. Although the long grain of ''Guardian'' results in a desirable high TGW the HLW is too low. To select mutants exhibiting increased HLW the character was first analyzed to identify traits that could more easily be screened for using M 2 seeds. In comparison of 6 wheat cultivars, correlation analyses with HLW resulted in coefficients of -0.86 (grain length, L:P 2 seeds for shorter, less prolate grains. Mutagenesis was carried out using EMS sulphonate (1.8 or 3.6%), sodium azide (2 or 20 mM) or X-rays (7.5 or 20 kR). 69 M 2 grains with altered shape were selected. Examination of the M 3 progeny confirmed 6 grain-shape mutants, most of them resulting from EMS treatment (Table). Two of the mutants showed TGW values significantly below the parental variety, but three mutants exhibited HLW and TGW values significantly greater than those of the parental variety. Microplot yield trails on selected M 3 lines are in progress. The influence of physical grain characteristics on HLW offers prospects for mechanical fractionation of large M 2 populations. The application of gravity separators (fractionation on the basis of grain density) and sieves (fractionation on the basis of grain length) in screening mutants possessing improved grain quality is being investigated
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Radiosynthesis and pre-clinical evaluation of [{sup 18}F]fluoro-[1,2-{sup 2}H{sub 4}]choline
Energy Technology Data Exchange (ETDEWEB)
Smith, Graham [Comprehensive Cancer Imaging Centre, Faculty of Medicine, Imperial College London, Hammersmith Hospital, Du Cane Road, London W12 0NN (United Kingdom); Zhao Yongjun [MDx Discovery (part of GE Healthcare) at Hammersmith Imanet, Ltd., Hammersmith Hospital, Du Cane Road, London W12 0NN (United Kingdom); Leyton, Julius [Comprehensive Cancer Imaging Centre, Faculty of Medicine, Imperial College London, Hammersmith Hospital, Du Cane Road, London W12 0NN (United Kingdom); Shan Bo [MDx Discovery (part of GE Healthcare) at Hammersmith Imanet, Ltd., Hammersmith Hospital, Du Cane Road, London W12 0NN (United Kingdom); Nguyen, Quang-de; Perumal, Meg [Comprehensive Cancer Imaging Centre, Faculty of Medicine, Imperial College London, Hammersmith Hospital, Du Cane Road, London W12 0NN (United Kingdom); Turton, David [GE-Imanet, Hammersmith Hospital, Du Cane Road, London, W12 0NN (United Kingdom); Arstad, Erik; Luthra, Sajinder K.; Robins, Edward G. [MDx Discovery (part of GE Healthcare) at Hammersmith Imanet, Ltd., Hammersmith Hospital, Du Cane Road, London W12 0NN (United Kingdom); Aboagye, Eric O., E-mail: eric.aboagye@imperial.ac.u [Comprehensive Cancer Imaging Centre, Faculty of Medicine, Imperial College London, Hammersmith Hospital, Du Cane Road, London W12 0NN (United Kingdom)
2011-01-15
Introduction: Choline radiotracers are widely used for clinical PET diagnosis in oncology. [{sup 11}C]Choline finds particular utility in the imaging of brain and prostate tumor metabolic status, where 2-[{sup 18}F]fluoro-2-deoxy-D-glucose ('FDG') shows high background uptake. More recently we have extended the clinical utility of [{sup 11}C]choline to breast cancer where radiotracer uptake correlates with tumor aggressiveness (grade). In the present study, a new choline analog, [{sup 18}F]fluoro-[1,2-{sup 2}H{sub 4}]choline, was synthesized and evaluated as a potential PET imaging probe. Methods: [{sup 18}F]Fluorocholine, [{sup 18}F]fluoro-[1-{sup 2}H{sub 2}]choline and [{sup 18}F]fluoro-[1,2-{sup 2}H{sub 4}]choline were synthesized by alkylation of the relevant precursor with [{sup 18}F]fluorobromomethane or [{sup 18}F]fluoromethyl tosylate. Radiosynthesis of [{sup 18}F]fluoromethyl tosylate required extensive modification of the existing method. [{sup 18}F]Fluorocholine and [{sup 18}F]fluoro-[1,2-{sup 2}H{sub 4}]choline were then subjected to in vitro oxidative stability analysis in a chemical oxidation model using potassium permanganate and an enzymatic model using choline oxidase. The two radiotracers, together with the corresponding di-deuterated compound, [{sup 18}F]fluoro-[1-{sup 2}H{sub 2}]choline, were then evaluated in vivo in a time-course biodistribution study in HCT-116 tumor-bearing mice. Results: Alkylation with [{sup 18}F]fluoromethyl tosylate proved to be the most reliable radiosynthetic route. Stability models indicate that [{sup 18}F]fluoro-[1,2-{sup 2}H{sub 4}]choline possesses increased chemical and enzymatic (choline oxidase) oxidative stability relative to [{sup 18}F]fluorocholine. The distribution of the three radiotracers, [{sup 18}F]fluorocholine, [{sup 18}F]fluoro-[1-{sup 2}H{sub 2}]choline and [{sup 18}F]fluoro-[1,2-{sup 2}H{sub 4}]choline, showed a similar uptake profile in most organs. Crucially, tumor uptake of [{sup 18}F
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Plant community development within the F- and H-Area tree-kill zone
Energy Technology Data Exchange (ETDEWEB)
Nelson, E.A.; Westbury, H.M. Jr.
1994-10-01
The F- and H-Area Seepage Basins received liquid waste from the F and H chemical separation facilities from 1955 through 1988. Tree mortality in seepline fed wetlands down-slope from the basins was observed in the late 1970`s, and investigations were conducted to determine the cause and source of the impacts. Analysis of the soil and water in the tree-kill zones demonstrated a strong chemical linkage with the F- and H-Area seepage basins. Although no single cause of the mortality was determined, it was believed to be the result of interactions of alterations in the hydrology and erosional deposition, along with lowering of pH and increased conductivity, sodium, aluminum, and nitrogen compounds. A mild drought during the growing season may also have increased the concentration of the chemical contaminants in the soils matrix. In 1988, the F- and H-Area Seepage Basins were closed and covered with a clay cap to reduce the rate of dispersion of the contaminants in the soil beneath the basins. Subsequent studies of the chemical composition of the tree-kill zone groundwater and toxicological characteristics of the seepline soil have shown a reduced contaminant flux. In 1993, an initial vegetation study was undertaken to determine the level of recovery by the plant communities in the tree-kill zones. This study repeats the initial vegetation investigation in order to further analyze and characterize the recovery of plant communities in the zones after an additional year of growth.
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Improved hydrogen storage properties of LiAlH4 by mechanical milling with TiF3
International Nuclear Information System (INIS)
Zang, Lei; Cai, Jiaxing; Zhao, Lipeng; Gao, Wenhong; Liu, Jian; Wang, Yijing
2015-01-01
Dehydrogenation behavior of LiAlH 4 (lithium alanate) admixed with TiF 3 is investigated by pressure-composition-temperature (PCT), fourier transform infrared spectroscopy (FTIR), X-ray Diffraction (XRD), differential scanning calorimetry (DSC) and temperature programmed desorption (TPD). The TiF 3 addition enhances kinetics of LiAlH 4 and decreases the decomposition temperature. The LiAlH 4 -2 mol % TiF 3 sample starts to release hydrogen at about 35 °C and the dehydrogenation rate reaches a maximum value at 108.4 °C, compared with 145 °C and 179.9 °C for the as-received LiAlH 4 . As for the dehydrogenation kinetics, the LiAlH 4 -2 mol % TiF 3 sample releases about 7.0 wt % H 2 at 140 °C within 80 min. In comparison, the as-received LiAlH 4 sample releases only 0.8 wt % hydrogen under the same conditions. The existence of proposed catalyst, Al 3 Ti formed in-situ in the process of dehydrogenation, has been confirmed experimentally by XRD measurements. The activation energy of LiAlH 4 -2 mol % TiF 3 composite is deduced to be 66.76 kJ mol −1 and 88.21 kJ mol −1 for the first and second reaction stages of LiAlH 4 dehydrogenation. - Highlights: • TiF 3 considerably enhances the dehydrogenation kinetics of LiAlH 4 . • TiF 3 -doped LiAlH 4 dehydrogenates even at room temperature. • Low activation energy of the dehydrogenation reaction. • Al 3 Ti formed in-situ helps to explain the dehydrogenation mechanism
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Friction stir welding of F82H steel for fusion applications
Energy Technology Data Exchange (ETDEWEB)
Noh, Sanghoon, E-mail: shnoh@kaeri.re.kr [Fusion Structural Materials Division, Japan Atomic Energy Agency, Rokkasho, Aomori (Japan); Nuclear Materials Division, Korea Atomic Energy Research Institute, Yuseong-gu, Daejeon (Korea, Republic of); Ando, Masami; Tanigawa, Hiroyasu [Fusion Structural Materials Division, Japan Atomic Energy Agency, Rokkasho, Aomori (Japan); Fujii, Hidetoshi [Joining and Welding Research Institute, Osaka University, Ibaraki, Osaka (Japan); Kimura, Akihiko [Institute of Advanced Energy, Kyoto University, Gokasho, Uji, Kyoto (Japan)
2016-09-15
In the present study, friction stir welding was employed to join F82H steels and develop a potential joining technique for a reduced activation ferritic/martensitic steel. The microstructures and mechanical properties on the joint region were investigated to evaluate the applicability of friction stir welding. F82H steel sheets were successfully butt-joined with various welding parameters. In welding conditions, 100 rpm and 100 mm/min, the stirred zone represented a comparable hardness distribution with a base metal. Stirred zone induced by 100 rpm reserved uniformly distributed precipitates and very fine ferritic grains, whereas the base metal showed a typical tempered martensite with precipitates on the prior austenite grain boundary and lath boundary. Although the tensile strength was decreased at 550 °C, the stirred zone treated at 100 rpm showed comparable tensile behavior with base metal up to 500 °C. Therefore, friction stir welding is considered a potential welding method to preserve the precipitates of F82H steel.
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Friction stir welding of F82H steel for fusion applications
International Nuclear Information System (INIS)
Noh, Sanghoon; Ando, Masami; Tanigawa, Hiroyasu; Fujii, Hidetoshi; Kimura, Akihiko
2016-01-01
In the present study, friction stir welding was employed to join F82H steels and develop a potential joining technique for a reduced activation ferritic/martensitic steel. The microstructures and mechanical properties on the joint region were investigated to evaluate the applicability of friction stir welding. F82H steel sheets were successfully butt-joined with various welding parameters. In welding conditions, 100 rpm and 100 mm/min, the stirred zone represented a comparable hardness distribution with a base metal. Stirred zone induced by 100 rpm reserved uniformly distributed precipitates and very fine ferritic grains, whereas the base metal showed a typical tempered martensite with precipitates on the prior austenite grain boundary and lath boundary. Although the tensile strength was decreased at 550 °C, the stirred zone treated at 100 rpm showed comparable tensile behavior with base metal up to 500 °C. Therefore, friction stir welding is considered a potential welding method to preserve the precipitates of F82H steel.
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Characterization of a Weak Allele of Zebrafish cloche Mutant
Ma, Ning; Huang, Zhibin; Chen, Xiaohui; He, Fei; Wang, Kun; Liu, Wei; Zhao, Linfeng; Xu, Xiangmin; Liao, Wangjun; Ruan, Hua; Luo, Shenqiu; Zhang, Wenqing
2011-01-01
Hematopoiesis is a complicated and dynamic process about which the molecular mechanisms remain poorly understood. Danio rerio (zebrafish) is an excellent vertebrate system for studying hematopoiesis and developmental mechanisms. In the previous study, we isolated and identified a cloche 172 (clo 172) mutant, a novel allele compared to the original cloche (clo) mutant, through using complementation test and initial mapping. Here, according to whole mount in-situ hybridization, we report that the endothelial cells in clo 172 mutant embryos, although initially developed, failed to form the functional vascular system eventually. In addition, further characterization indicates that the clo 172 mutant exhibited weaker defects instead of completely lost in primitive erythroid cells and definitive hematopoietic cells compared with the clo s5 mutant. In contrast, primitive myeloid cells were totally lost in clo 172 mutant. Furthermore, these reappeared definitive myeloid cells were demonstrated to initiate from the remaining hematopoietic stem cells (HSCs) in clo 172 mutant, confirmed by the dramatic decrease of lyc in clo 172 runx1w84x double mutant. Collectively, the clo 172 mutant is a weak allele compared to the clo s5 mutant, therefore providing a model for studying the early development of hematopoietic and vascular system, as well as an opportunity to further understand the function of the cloche gene. PMID:22132109
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Vattanaviboon, Paiboon; Whangsuk, Wirongrong; Mongkolsuk, Skorn
2003-01-01
We isolated menadione-resistant mutants of Xanthomonas campestris pv. phaseoli oxyR (oxyRXp). The oxyRR2Xp mutant was hyperresistant to the superoxide generators menadione and plumbagin and was moderately resistant to H2O2 and tert-butyl hydroperoxide. Analysis of enzymes involved in oxidative-stress protection in the oxyRR2Xp mutant revealed a >10-fold increase in AhpC and AhpF levels, while the levels of superoxide dismutase (SOD), catalase, and the organic hydroperoxide resistance protein (Ohr) were not significantly altered. Inactivation of ahpC in the oxyRR2Xp mutant resulted in increased sensitivity to menadione killing. Moreover, high levels of expression of cloned ahpC and ahpF in the oxyRXp mutant complemented the menadione hypersensitivity phenotype. High levels of other oxidant-scavenging enzymes such as catalase and SOD did not protect the cells from menadione toxicity. These data strongly suggest that the toxicity of superoxide generators could be mediated via organic peroxide production and that alkyl hydroperoxide reductase has an important novel function in the protection against the toxicity of these compounds in X. campestris. PMID:12591894
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An accurate potential energy surface for the F + H2 → HF + H reaction by the coupled-cluster method
International Nuclear Information System (INIS)
Chen, Jun; Sun, Zhigang; Zhang, Dong H.
2015-01-01
A three dimensional potential energy surface for the F + H 2 → HF + H reaction has been computed by the spin unrestricted coupled cluster method with singles, doubles, triples, and perturbative quadruples [UCCSDT(2) Q ] using the augmented correlation-consistent polarised valence quadruple zeta basis set for the fluorine atom and the correlation-consistent polarised valence quadruple zeta basis set for the hydrogen atom. All the calculations are based on the restricted open-shell Hartree-Fock orbitals, together with the frozen core approximations, and the UCCSD(T)/complete basis set (CBS) correction term was included. The global potential energy surface was calculated by fitting the sampled ab initio points without any scaling factor for the correlation energy part using a neutral network function method. Extensive dynamics calculations have been carried out on the potential energy surface. The reaction rate constants, integral cross sections, product rotational states distribution, and forward and backward scattering as a function of collision energy of the F + HD → HF + D, F + HD → DF + H, and F + H 2 reaction, were calculated by the time-independent quantum dynamics scattering theory using the new surface. The satisfactory agreement with the reported experimental observations previously demonstrates the accuracy of the new potential energy surface
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Directory of Open Access Journals (Sweden)
Sujuan Chen
2017-06-01
Full Text Available H9N2 avian influenza virus is a zoonotic agent with a broad host range that can contribute genetic information to H5 or H7N9 subtype viruses, which are significant threats to both humans and birds. Thus, there is a great need for a vaccine to control H9N2 avian influenza. Three mutant viruses of an H9N2 virus A/chicken/Taixing/10/2010 (rTX-NS1-73, rTX-NS1-100, and rTX-NS1-128 were constructed with different NS1 gene truncations and confirmed by western blot analysis. The genetic stability, pathogenicity, transmissibility, and host immune responses toward these mutants were evaluated. The mutant virus rTX-NS1-128 exhibited the most attenuated phenotype and lost transmissibility. The expression levels of interleukin 12 in the nasal and tracheal tissues from chickens immunized with rTX-NS1-128 were significantly upregulated on day 3 post-immunization and the IgA and IgG antibody levels were significantly increased on days 7, 14, and 21 post-immunization when compared to chickens that received an inactivated vaccine. rTX-NS1-128 also protected chickens from challenge by homologous and heterologous H9N2 avian influenza viruses. The results indicate that rTX-NS1-128 can be used as a potential live-attenuated vaccine against H9N2 avian influenza.
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Results of the F/H Effluent Treatment Facility biological monitoring program, July 1987--July 1991
International Nuclear Information System (INIS)
Specht, W.L.
1992-07-01
As required by the South Carolina Department of Health and Environmental Control (SCDHEC) under NPDES Permit SCO000175, biological monitoring was conducted in Upper Three Runs Creek to determine if discharges from the F/H Effluent Treatment Facility have adversely impacted the biotic community of the receiving stream. Data included in this summary report encompass July 1987 through July 1991. As originally designed, the F/H ETF was not expected to remove all of the mercury from the wastewater; therefore, SCDHEC specified that studies be conducted to determine if mercury was bioaccumulating in aquatic biota. Subsequent to approval of the biological monitoring program, an ion exchange column was added to the F/H ETF specifically to remove mercury, which eliminated mercury from the F/H ETF effluent. The results of the biological monitoring program indicate that at the present rate of discharge, the F/H ETF effluent has not adversely affected the receiving stream with respect to any of the parameters that were measured. The effluent is not toxic at the in-stream waste concentration and there is no evidence of mercury bioaccumulation
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Jean-Claude Gadmer
2012-01-01
CERN-HI-1203078 01 - 13: visit of LINAC 3 with M. Benedikt CERN-HI-1203078 14 - 20: in the Roy Billinge room CERN-HI-1203078 21- 22: visit of the LEIR accelerator in building 354 CERN-HI-1203078 23 - 55: signing the guest book with CERN Director-General R. Heuer and Head of International Relations F. Pauss CERN-HI-1203078 56 - 99: in building 184, Governor Pröll and CERN Director-General R. Heuer switch on the MedAustron ion source to produce the proton beam; visit MedAustron facility. CERN-HI-1203078 32:from left to right: Klubobmann and Chairman of EBG MedAustron GmbH Council K. Schneeberger; Director-General R. Heuer;Niederösterreich Governor E. Pröll;Head of International Relations F. Pauss;Permanent Representative of Austria to the UNO and Austrian Delegate to CERN Council Ambassador C. Strohal.
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Pärt: Collage sur B-A-C-H für Kammerorschester / Hans-Christian Dadelsen
Dadelsen, Hans-Christian
1993-01-01
Uuest heliplaadist "Pärt: Collage sur B-A-C-H für Kammerorschester, Summa (1991) für Streichorchester, Fratres, Sinfonie Nr. 2, Festina lente, Wenn Bach Bienen gezüchtet hätte, Credo für Klavier, Chor und Orchester. Philharmonia Orchestra and Chorus, Neeme Järvi". Chandos/Koch CD 9134 (WD: 63'02")
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Fluorometric assay for phenotypic differentiation of drug-resistant HIV mutants
Zhu, Qinchang; Yu, Zhiqiang; Kabashima, Tsutomu; Yin, Sheng; Dragusha, Shpend; El-Mahdy, Ahmed F. M.; Ejupi, Valon; Shibata, Takayuki; Kai, Masaaki
2015-01-01
Convenient drug-resistance testing of viral mutants is indispensable to effective treatment of viral infection. We developed a novel fluorometric assay for phenotypic differentiation of drug-resistant mutants of human immunodeficiency virus-I protease (HIV-PR) which uses enzymatic and peptide-specific fluorescence (FL) reactions and high-performance liquid chromatography (HPLC) of three HIV-PR substrates. This assay protocol enables use of non-purified enzyme sources and multiple substrates f...
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Allosteric Mutant IDH1 Inhibitors Reveal Mechanisms for IDH1 Mutant and Isoform Selectivity
Energy Technology Data Exchange (ETDEWEB)
Xie, Xiaoling; Baird, Daniel; Bowen, Kimberly; Capka, Vladimir; Chen, Jinyun; Chenail, Gregg; Cho, YoungShin; Dooley, Julia; Farsidjani, Ali; Fortin, Pascal; Kohls, Darcy; Kulathila, Raviraj; Lin, Fallon; McKay, Daniel; Rodrigues, Lindsey; Sage, David; Touré, B. Barry; van der Plas, Simon; Wright, Kirk; Xu, Ming; Yin, Hong; Levell, Julian; Pagliarini, Raymond A. (Novartis)
2017-03-01
Oncogenic IDH1 and IDH2 mutations contribute to cancer via production of R-2-hydroxyglutarate (2-HG). Here, we characterize two structurally distinct mutant- and isoform-selective IDH1 inhibitors that inhibit 2-HG production. Both bind to an allosteric pocket on IDH1, yet shape it differently, highlighting the plasticity of this site. Oncogenic IDH1R132H mutation destabilizes an IDH1 “regulatory segment,” which otherwise restricts compound access to the allosteric pocket. Regulatory segment destabilization in wild-type IDH1 promotes inhibitor binding, suggesting that destabilization is critical for mutant selectivity. We also report crystal structures of oncogenic IDH2 mutant isoforms, highlighting the fact that the analogous segment of IDH2 is not similarly destabilized. This intrinsic stability of IDH2 may contribute to observed inhibitor IDH1 isoform selectivity. Moreover, discrete residues in the IDH1 allosteric pocket that differ from IDH2 may also guide IDH1 isoform selectivity. These data provide a deeper understanding of how IDH1 inhibitors achieve mutant and isoform selectivity.
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Mildew resistant and less lodging wheat mutants induced in Iran
International Nuclear Information System (INIS)
Naghedi-Ahmadi, I.
1989-01-01
''Tabassi'' is a lodging and mildew susceptible cultivar. To induce mutations, seeds were gamma irradiated (50 to 150 Gy) in 1982 and selection for lodging resistance was carried out in M 2 . During field experiments with the mutant lines in 1985/86 there has been a heavy mildew epidemic under which mutant 63-5-I (derived from 50 Gy treatment) exhibited considerable resistance and as a consequence, higher yield. The control was 100% infected, the mutant only 40%. The mutant yielded 31% more grain, 7.5% less straw and 4.5% more protein than the control. Lodging of 63-5-I was only 60% in an experiment under rainfed conditions in the same season, resulting in a relative yield increase of about 11%. In 1986/87 there was no mildew epidemic and the mutant yielded the same as ''Tabassi''
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Directory of Open Access Journals (Sweden)
Matthew J. Montgomery
2015-07-01
Full Text Available The two title compounds are isomers of C6H3ClN2 containing a pyridine ring, a nitrile group, and a chloro substituent. The molecules of each compound pack together in the solid state with offset face-to-face π-stacking, and intermolecular C—H...Nnitrile and C—H...Npyridine interactions. 4-Chloropyridine-2-carbonitrile, (I, exhibits pairwise centrosymmetric head-to-head C—H...Nnitrile and C—H...Npyridine interactions, forming one-dimensional chains, which are π-stacked in an offset face-to-face fashion. The intermolecular packing of the isomeric 6-chloropyridine-2-carbonitrile, (II, which differs only in the position of the chloro substituent on the pyridine ring, exhibits head-to-tail C—H...Nnitrile and C—H...Npyridine interactions, forming two-dimensional sheets which are π-stacked in an offset face-to-face fashion. In contrast to (I, the offset face-to-face π-stacking in (II is formed between molecules with alternating orientations of the chloro and nitrile substituents.
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International Nuclear Information System (INIS)
Zhang, J.; Huang, Y.N.; Mao, C.; Peng, P.
2012-01-01
Highlights: ► The co-incorporation of Ti and F into MgH 2 lattice is energetically favorable. ► The incorporated Ti and F in MgH 2 preferably generate TiH 2 and MgF 2 , respectively. ► The synergistic effect of Ti and F is superior to that of pure Ti. ► The weakened interactions of Mg–H explain enhanced dehydrogenation properties. - Abstract: The energetic and electronic properties of MgH 2 co-doped with Ti and F are investigated using first-principles calculations based on density functional theory. The calculation results show that incorporation of Ti combined with F atoms into MgH 2 lattice is energetically favorable relative to single incorporation of Ti atom. After dehydrogenation, the co-doped Ti and F in MgH 2 preferably generate TiH 2 and MgF 2 , respectively. Comparatively, the combined effect of Ti and F in improving the dehydrogenation properties of MgH 2 is superior to that of pure Ti. These results provide a reasonable explanation for experimental observations. Analysis of electronic structures suggests the enhanced dehydrogenation properties of doped MgH 2 can be attributed to the weakened bonding interactions between Mg and H due to foreign species doping.
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An early maturing rice mutant released as a variety
International Nuclear Information System (INIS)
Azam, M.A.; Imtiaz Uddin, Md.
2001-01-01
In the content of food grain production deficiency (about 1.0-1.5 million tons of rice per year according to the Bangladesh Bureau of Statistics, 1998) an induced mutation programme was undertaken in 1985. One moderate early maturing and high yielding rice mutant line (BINA6-84-4-115) has been developed by irradiating F 2 seeds of the cross 'BR4' x 'Iratom 38'. Three treatments viz., 250, 300 and 350 Gy were given to the F 2 seeds. Finally, this line was selected in M 6 generation for advanced yield trial. The line was evaluated in comparative trials with another mutant line BINA6-84-4-163. These two mutant lines had been selected earlier from 300 Gy originated lines. The two check varieties, 'BR 11' and 'BR 22' were also included in the trial, which was conducted in two consecutive T. aman seasons (July to December) during 1994 and 1995 at five locations in Bangladesh. From the results, it was evident that the mutant BINA6-84-4-115 did not differ much with the other mutant lines or check varieties in respect to plant height, number of effective tillers and panicle length but it was 10-18 days earlier than the other 3 entries. It produced a similar yield as the check BR 11 in 1994 and a higher yield than the check BR 11 and BR 22 in 1995. This mutant line gave the highest yield per day among all the entries. In addition to this, the grains are long, fine and possess a high L/B ratio, which are of high commercial value. This line has been released by the National Seed Board of Bangladesh in 1998 as a commercial variety under the name 'BINADHAN-4' for cultivation throughout Bangladesh
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Photo dynamics of BLUF domain mutant H44R of AppA from Rhodobacter sphaeroides
Energy Technology Data Exchange (ETDEWEB)
Zirak, P. [Institut II - Experimentelle und Angewandte Physik, Universitaet Regensburg, Universitaetstrasse 31, D-93053 Regensburg (Germany); Penzkofer, A. [Institut II - Experimentelle und Angewandte Physik, Universitaet Regensburg, Universitaetstrasse 31, D-93053 Regensburg (Germany)], E-mail: alfons.penzkofer@physik.uni-regensburg.de; Hegemann, P.; Mathes, T. [Institut fuer Biologie, Experimentelle Biophysik, Humboldt-Universitaet zu Berlin, Invalidenstr. 42, D-10115 Berlin (Germany)
2007-05-21
The photo-cycle dynamics of the H44R mutant of the BLUF domain of the transcriptional anti-repressor protein AppA (AppA-H44R) from the non-sulfur anoxyphototropic purple bacterium Rhodobacter sphaeroides is studied in order to gain information on the involvement of His44 in the photo-cyclic mechanism of the AppA BLUF domain and to add information to the involved processes. The amino acid residue histidine at position 44 is replaced by arginine. A 12 nm red-shifted signalling state is formed upon blue-light excitation, while in wild-type AppA (AppA-wt) the red-shift is 16 nm. The recovery to the receptor dark state is approximately a factor of 2.5 faster ({tau}{sub rec} {approx} 6.5 min) than the recovery of the wild-type counterpart. Extended light exposure of the mutant causes photo-degradation of flavin (mainly free flavin conversion to lumichrome and re-equilibration between free and non-covalently bound flavin) and protein aggregation (showing up as light scattering). No photo-degradation was observed for AppA-wt. The quantum efficiency of signalling-state formation determined by intensity dependent absorption measurements is found to be {phi}{sub s} {approx} 0.3 (for AppA-wt: {phi}{sub s} {approx} 0.24). A two-component single-exponential fluorescence relaxation was observed, which is interpreted as fast fluorescence quenching to an equilibrium value by photo-induced electron transfer followed by slower fluorescence decay due to charge recombination. Based on the experimental findings, an extended photo-cycle model for BLUF domains is proposed.
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International Nuclear Information System (INIS)
Boyd, J.B.; Golino, M.D.; Nguyen, T.D.; Green, M.M.
1976-01-01
Thirteen X-linked mutants have been isolated in Drosophila melanogaster which render male and homozygous female larvae sensitive to the mutagen methyl methanesulfonate. Their characterization and preliminary assignment to functional groups is described. Four of these mutants are alleles of mei-41. Like previously isolated alleles of this locus, these mutants reduce fertility and increase loss and nondisjunction of the X-chromosome in homozygous females. The remaining mutants have been tentatively assigned to six functional groups (two mutants to the mus(1)101 locus, two to mus(1)102, two to mus(1)103, and one each to mus(1)104, mus(1)105, and mus(1)106. Several of the complementation groups can be distinguished on the basis of nondisjunction and cross sensitivity to mutagens. Females homozygous for the mei-41, mus(1)101 and mus(1)102 mutants exhibit elevated levels of nondisjunction. Mutants belonging to complementation groups mei-41, mus(1)101, and mus(1)104 are sensitive to nitrogen mustard (HN2) in addition to their MMS sensitivity. Among these mutants there is currently a direct correlation between sensitivity to HN2, sensitivity to 2-acetylaminofluorene and a deficiency in post-replication repair. Only the mei-41 mutants are hypersensitive to uv radiation, although several of the mutants exhibit sensitivity to γ-rays. Semidominance is observed in female larvae of the mei-41, mus(1)104, and mus(1)103 mutants after exposure to high concentrations of MMS. The properties of the mutants generally conform to a pattern which has been established for related mutants in yeast
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Intracellular distribution of a speech/language disorder associated FOXP2 mutant
International Nuclear Information System (INIS)
Mizutani, Akifumi; Matsuzaki, Ayumi; Momoi, Mariko Y.; Fujita, Eriko; Tanabe, Yuko; Momoi, Takashi
2007-01-01
Although a mutation (R553H) in the forkhead box (FOX)P2 gene is associated with speech/language disorder, little is known about the function of FOXP2 or its relevance to this disorder. In the present study, we identify the forkhead nuclear localization domains that contribute to the cellular distribution of FOXP2. Nuclear localization of FOXP2 depended on two distally separated nuclear localization signals in the forkhead domain. A truncated version of FOXP2 lacking the leu-zip, Zn 2+ finger, and forkhead domains that was observed in another patient with speech abnormalities demonstrated an aggregated cytoplasmic localization. Furthermore, FOXP2 (R553H) mainly exhibited a cytoplasmic localization despite retaining interactions with nuclear transport proteins (importin α and β). Interestingly, wild type FOXP2 promoted the transport of FOXP2 (R553H) into the nucleus. Mutant and wild type FOXP2 heterodimers in the nucleus or FOXP2 R553H in the cytoplasm may underlie the pathogenesis of the autosomal dominant speech/language disorder
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Temperature-sensitive host range mutants of herpes simplex virus type 2
International Nuclear Information System (INIS)
Koment, R.W.; Rapp, F.
1975-01-01
Herpesviruses are capable of several types of infection of a host cell. To investigate the early events which ultimately determine the nature of the virus-host cell interaction, a system was established utilizing temperature-sensitive mutants of herpes simplex virus type 2. Four mutants have been isolated which fail to induce cytopathic effects and do not replicate at 39 C in hamster embryo fibroblast cells. At least one mutant is virus DNA negative. Since intracellular complementation is detectable between pairs of mutants, a virus function is known to be temperature sensitive. However, all four mutants induce cytopathic effects and replicate to parental virus levels in rabbit kidney cells at 39 C. This suggests that a host cell function, lacking or nonfunctional in HEF cells but present in rabbit kidney cells at 39 C, is required for the replication of these mutants in hamster embryo fibroblast cells at 39 C. Therefore, we conclude that these mutants are both temperature sensitive and exhibit host range properties
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International Nuclear Information System (INIS)
Mao Dezhi; Tang Yilan
1998-01-01
The hybrid rice F 1 plant derived from the back crossing of glutinous rice x wild rice was irradiated with γ-ray. The result of investigation to the induced mutant showed that through the selection and backcross, a black glutinous rice strain with the short stem, cold tolerance and high yield was developed. The analysis of the ability of heredity variance showed that the selection was effective for the husk colour, black glutinous and black Indica rice, but ineffective for the white Indica rice and seed setting
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Serrated leaf mutant in mungbean (Vigna radiata (L) Wilczek)
International Nuclear Information System (INIS)
Malik, I.A.; Ghulam, Sarwar; Yousaf, Ali; Saleem, M.
1988-01-01
Dry dormant seeds of mungbean (Vigna radiata (L) Wilczek) were treated with gamma rays (15, 30 and 60 kR). The serrated leaf mutation was noticed in M 2 of cultivar Pak 32 treated with 60 kR. Cf 14 plants, 3 showed the altered leaf structure and the others were normal. The feature of this mutant was the deep serration of leaflet margins. The mutant had large thick leaflets with prominent venation. The mutant bred true in the M 3 and successive generation. Details of the morphological characteristics of the mutant are presented. The mutant exhibited slower growth particularly during the early stages of development, flowered later and attained shorter height. There was an increase in the number of pods, in seed weight and in seed protein content, but number of seed per pod was considerably reduced. The seed coat colour showed a change from green to yellowish green. In the mutant's flowers the stamina were placed much below the stigma level and the stigma sometimes protruded the corolla. Outcrossing of 4% recorded in some of the mutant lines revealed a reduced cleistogamy. The low number of seeds per pod in the mutant could be due to reduced pollen fertility. The mutant behaved as monogenic recessive. The symbols SL/sl are proposed for this allelic pair. The mutant may have use as a green manure crop because of its large foliage and for the breeders as a genetic marker
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DEFF Research Database (Denmark)
Christensen, Lise Lotte; Bross, Peter Gerd; Ørntoft, Torben Falck
2000-01-01
Human alpha1,3-fucosyltransferase V and -VI (hFucTV and -VI) each contain four potential N-glycosylation sites (hFucTV: Asn60, Asn105, Asn167 and Asn198 and hFucTVI: Asn46, Asn91, Asn153 and Asn184). Glycosylation of the two N-terminal potential N-glycosylation sites (hFucTV: Asn60, Asn105 and h......FucTVI: Asn46 and Asn91) have never been studied in detail. In the present study, we have analysed the glycosylation of these potential N-glycosylation sites. Initially, we compared the molecular mass of hFucTV and -VI expressed in COS-7 cells treated with tunicamycin with the mass of the proteins...... in untreated cells. The difference in molecular mass between the proteins in treated and untreated cells corresponded to the presence of at least three N-linked glycans. We then made a series of mutants, in which the asparagine residues in the N-terminal potential N-glycosylation sites were replaced...
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Structural transitions, ion mobility, and conductivity in CsSbF3(H2PO4)
Kavun, V. Ya.; Uvarov, N. F.; Slobodyuk, A. B.; Ulihin, A. S.; Kovaleva, E. V.; Zemnukhova, L. A.
2018-02-01
Structural transitions, ion mobility, and conductivity in CsSbF3(H2PO4) (I) have been investigated by the methods of 1H, 19F, 31P NMR (including 1H, 19F, 31P MAS NMR), DSC, X-ray diffraction, and impedance spectroscopy. It was found that the fundamental changes in 1H, 19F, 31P NMR spectra (above 390 K) were associated with the formation of a crystalline disorder phase I with high ionic mobility in the proton and fluoride sublattices, as a result of a phase transition in the 400-420 K range. In the same temperature range, the transition of PO2(OH)2- anions from the "rigid lattice" to fast reorientations takes place. Above 430 K, there occurs a transition from the crystalline disordered phase to the amorphous one. The types of ion mobility in CsSbF3(H2PO4) and its amorphous phase have been established and temperature ranges of their realization have been determined (150-450 K). According to the NMR data, the diffusion in the proton sublattice of the disordered crystalline and amorphous phases is preserved even at room temperature. The ionic conductivity in CsSbF3(H2PO4) reaches the values of 2.6 × 10-4 S/cm in the temperature range 410-425 K and decreases down to 2.0 × 10-5 S/cm upon transition to the amorphous phase (435-445 K).
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Tani, Tatsunori; Taguchi, Hisataka; Fujimori, Kazuhiro E; Sahara, Takehiko; Ohgiya, Satoru; Kamagata, Yoichi; Akamatsu, Takashi
2016-10-01
To clarify the mechanisms of xylitol utilization, three xylitol-assimilating mutants were isolated from recombinant Saccharomyces cerevisiae strains showing highly efficient xylose-utilization. The nucleotide sequences of the mutant genomes were analyzed and compared with those of the wild-type strains and the mutation sites were identified. gal80 mutations were common to all the mutants, and recessive to the wild-type allele. Hence we constructed a gal80Δ mutant and confirmed that the gal80Δ mutant showed a xylitol-assimilation phenotype. When the constructed gal80Δ mutant was crossed with the three isolated mutants, all diploid hybrids showed xylitol assimilation, indicating that the mutations were all located in the GAL80. We analyzed the role of the galactose permease Gal2, controlled by the regulatory protein Gal80, in assimilating xylitol. A gal2Δ gal80Δ double mutant did not show xylitol assimilation, whereas expression of GAL2 under the control of the TDH3 promoter in the GAL80 strain did result in assimilation. These data indicate that Gal2 was needed for xylitol assimilation in the wild-type strain. When the gal80 mutant with an initial cell concentration of A660 = 20 was used for batch fermentation in a complex medium containing 20 g/L xylose or 20 g/L xylitol at pH 5.0 and 30°C under oxygen limitation, the gal80 mutant consumed 100% of the xylose within 12 h, but xylitol within 100 h, indicating that xylose reductase is required for xylitol consumption in oxygen-limited conditions. Copyright © 2016 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.
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Electric field effect on the emission rate of H4F and H4S hole traps in InP
International Nuclear Information System (INIS)
Darwich, R.; Alek, B.
2010-01-01
The electric field effect on the emission rate enhancement of the H4 F and H4 S hole trap in highly Zn-doped InP has been examined using the deep level transient spectroscopy (DLTS) and double correlation DLTS (DDLTS). The DLTS and DDLTS results have been found to be in good agreement for low and intermediate electric fields, but they disagree for large field effect. Comparing our emission data with the theory, we have found that H4 F obeys the quantum model of phonon-assisted tunneling while H4 S follows the Poole-Frenkel model employing a three-dimensional screening coulombic potential. Our results show that the H4 S defect can be attributed to a charged (V p - Zn) complex. (author)
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International Nuclear Information System (INIS)
Hoy, C.A.; Thompson, L.H.; Mooney, C.L.; Salazar, E.P.
1985-01-01
DNA repair-deficient mutants from five genetic complementation groups isolated previously from Chinese hamster cells were assayed for survival after exposure to the bifunctional alkylating agents mitomycin C or diepoxybutane. Groups 1, 3, and 5 exhibited 1.6- to 3-fold hypersensitivity compared to the wild-type cells, whereas Groups 2 and 4 exhibited extraordinary hypersensitivity. Mutants from Groups 1 and 2 were exposed to 22 other bifunctional alkylating agents in a rapid assay that compared cytotoxicity of the mutants to the wild-type parental strain, AA8. With all but two of the compounds, the Group 2 mutant (UV4) was 15- to 60-fold more sensitive than AA8 or the Group 1 mutant (UV5). UV4 showed only 6-fold hypersensitivity to quinacrine mustard. Alkaline elution measurements showed that this compound produced few DNA interstrand cross-links but numerous strand breaks. Therefore, the extreme hypersensitivity of mutants from Groups 2 and 4 appeared specific for compounds the main cytotoxic lesions of which were DNA cross-links. Mutant UV5 was only 1- to 4-fold hypersensitive to all the compounds. Although the initial number of cross-links was similar for the three cell lines, the efficiency of removal of cross-links was lowest in UV4 and intermediate in UV5. These results suggest that the different levels of sensitivity are specifically related to different efficiencies of DNA cross-link removal. The phenotype of hypersensitivity to both UV radiation and cross-link damage exhibited by the mutants in Groups 2 and 4 appears to differ from those of the known human DNA repair syndromes
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Karremann, Michael; Gielen, Gerrit H; Hoffmann, Marion; Wiese, Maria; Colditz, Niclas; Warmuth-Metz, Monika; Bison, Brigitte; Claviez, Alexander; van Vuurden, Dannis G; von Bueren, André O; Gessi, Marco; Kühnle, Ingrid; Hans, Volkmar H; Benesch, Martin; Sturm, Dominik; Kortmann, Rolf-Dieter; Waha, Andreas; Pietsch, Torsten; Kramm, Christof M
2018-01-10
The novel entity of "diffuse midline glioma, H3 K27M-mutant" has been defined in the 2016 revision of the World Health Organization (WHO) classification of tumors of the central nervous system (CNS). Tumors of this entity arise in CNS midline structures of predominantly pediatric patients and are associated with an overall dismal prognosis. They are defined by K27M mutations in H3F3A or HIST1H3B/C, encoding for histone 3 variants H3.3 and H3.1, respectively, which are considered hallmark events driving gliomagenesis. Here, we characterized 85 centrally reviewed diffuse gliomas on midline locations enrolled in the nationwide pediatric German HIT-HGG registry regarding tumor site, histone 3 mutational status, WHO grade, age, sex, and extent of tumor resection. We found 56 H3.3 K27M-mutant tumors (66%), 6 H3.1 K27M-mutant tumors (7%), and 23 H3-wildtype tumors (27%). H3 K27M-mutant gliomas shared an aggressive clinical course independent of their anatomic location. Multivariate regression analysis confirmed the significant impact of the H3 K27M mutation as the only independent parameter predictive of overall survival (P = 0.009). In H3 K27M-mutant tumors, neither anatomic midline location nor histopathological grading nor extent of tumor resection had an influence on survival. These results substantiate the clinical significance of considering diffuse midline glioma, H3 K27M-mutant, as a distinct entity corresponding to WHO grade IV, carrying a universally fatal prognosis. © The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com
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21 CFR 184.1387 - Lactase enzyme preparation from Candida pseudotropicalis.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Lactase enzyme preparation from Candida pseudotropicalis. 184.1387 Section 184.1387 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND... preparation from Candida pseudotropicalis. (a) This enzyme preparation is derived from the nonpathogenic...
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E2F1 interactions with hHR23A inhibit its degradation and promote DNA repair.
Singh, Randeep K; Dagnino, Lina
2016-05-03
Nucleotide excision repair (NER) is a major mechanism for removal of DNA lesions induced by exposure to UV radiation in the epidermis. Recognition of damaged DNA sites is the initial step in their repair, and requires multiprotein complexes that contain XPC and hHR23 proteins, or their orthologues. A variety of transcription factors are also involved in NER, including E2F1. In epidermal keratinocytes, UV exposure induces E2F1 phosphorylation, which allows it to recruit various NER factors to sites of DNA damage. However, the relationship between E2F1 and hHR23 proteins vis-à-vis NER has remained unexplored. We now show that E2F1 and hHR23 proteins can interact, and this interaction stabilizes E2F1, inhibiting its proteasomal degradation. Reciprocally, E2F1 regulates hHR23A subcellular localization, recruiting it to sites of DNA photodamage. As a result, E2F1 and hHR23A enhance DNA repair following exposure to UV radiation, contributing to genomic stability in the epidermis.
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International Nuclear Information System (INIS)
Koch, J.W.; Dixon, K.L.
1995-01-01
The Environmental Sciences Section of the Savannah River Technology Center established a quarterly monitoring program of the Fourmile Branch stream and its associated seepline located down gradient from the F- and H-Area Seepage Basins. The primary focus of this program was to survey and track changes in tritium levels; however, specific conductivity, and pH were also surveyed and tracked. The measurement from the eleventh survey (May 1995) exhibited similar tritium levels, conductivity measurements, and pH values to data from previous sampling events. The overall results of the tritium survey and stream monitoring data (Looney et al., 1993) indicate that the tritium plume resulting from the past operation of the seepage basins continues to flush from the Fourmile Branch wetland system
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Indian Academy of Sciences (India)
Home; Journals; Bulletin of Materials Science. F F Sun. Articles written in Bulletin of Materials Science. Volume 37 Issue 1 February 2014 pp 71-76. Study of electroless copper plating on ABS resin surface modified by heterocyclic organosilane self-assembled film · H N Zhang J Wang F F Sun D Liu H Y Wang F Wang.
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Directory of Open Access Journals (Sweden)
Muhammad Ibrahim Rajoka
2011-01-01
Full Text Available Kinetics of intracellular invertase production employing a double mutant derivative of Kluyveromyces marxianus was optimized by varying different process variables in a 23-litre fermentor. The maximum volumetric rate (QP and invertase yield (YP/S by M15 mutant were 1222 U/(L·h and 160 U/g of substrate utilized, respectively (2-fold more than those of parental strain at 50 °C on the molasses (150 g/L of total fermentable sugars at pH=5.5. Glucose or sucrose (100, 150 or 170 g/L did not repress invertase catabolically under the optimized fermentation conditions, contrary to the previous reports on other yeasts and filamentous fungi, where catabolite repression of sugars was predominant. Invertases derived by the wild (IW and mutant (IM strains were purified employing ammonium sulphate precipitation, and then characterized by column chromatographic techniques both kinetically and thermodynamically. The acidic limb of invertases was missing and collation of pKa and the heat of ionization values indicated that carboxyl groups were involved in proton transfer during active catalysis. Ratios of Kcat/Km and vmax/Km indicated that IM was significantly more specific for sucrose hydrolysis. The IM exhibited stability in different buffers at pH=3.0–10.0 and temperature of 50–70 °C, as reflected by long half-lives. IM showed significantly lower values of enthalpy of activation (ΔH* and entropy of activation (ΔS*, while Gibbs free energy (ΔG* was significantly increased at higher temperatures, making the IM thermodynamically more thermostable. Thus IM could be used as a catabolite-resistant invertase for the production of fructose syrup or high gravity ethanol.
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Energy Technology Data Exchange (ETDEWEB)
Chen, Jun; Sun, Zhigang, E-mail: zsun@dicp.ac.cn, E-mail: zhangdh@dicp.ac.cn; Zhang, Dong H., E-mail: zsun@dicp.ac.cn, E-mail: zhangdh@dicp.ac.cn [State Key Laboratory of Molecular Reaction Dynamics and Center for Theoretical Computational Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023 (China)
2015-01-14
A three dimensional potential energy surface for the F + H{sub 2} → HF + H reaction has been computed by the spin unrestricted coupled cluster method with singles, doubles, triples, and perturbative quadruples [UCCSDT(2){sub Q}] using the augmented correlation-consistent polarised valence quadruple zeta basis set for the fluorine atom and the correlation-consistent polarised valence quadruple zeta basis set for the hydrogen atom. All the calculations are based on the restricted open-shell Hartree-Fock orbitals, together with the frozen core approximations, and the UCCSD(T)/complete basis set (CBS) correction term was included. The global potential energy surface was calculated by fitting the sampled ab initio points without any scaling factor for the correlation energy part using a neutral network function method. Extensive dynamics calculations have been carried out on the potential energy surface. The reaction rate constants, integral cross sections, product rotational states distribution, and forward and backward scattering as a function of collision energy of the F + HD → HF + D, F + HD → DF + H, and F + H{sub 2} reaction, were calculated by the time-independent quantum dynamics scattering theory using the new surface. The satisfactory agreement with the reported experimental observations previously demonstrates the accuracy of the new potential energy surface.
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Disturbed secretion of mutant adiponectin associated with the metabolic syndrome.
Kishida, Ken; Nagaretani, Hiroyuki; Kondo, Hidehiko; Kobayashi, Hideki; Tanaka, Sachiyo; Maeda, Norikazu; Nagasawa, Azumi; Hibuse, Toshiyuki; Ohashi, Koji; Kumada, Masahiro; Nishizawa, Hitoshi; Okamoto, Yoshihisa; Ouchi, Noriyuki; Maeda, Kazuhisa; Kihara, Shinji; Funahashi, Tohru; Matsuzawa, Yuji
2003-06-20
Adiponectin, an adipocyte-derived protein, consists of collagen-like fibrous and complement C1q-like globular domains, and circulates in human plasma in a multimeric form. The protein exhibits anti-diabetic and anti-atherogenic activities. However, adiponectin plasma concentrations are low in obese subjects, and hypoadiponectinemia is associated with the metabolic syndrome, which is a cluster of insulin resistance, type 2 diabetes mellitus, hypertension, and dyslipidemia. We have recently reported a missense mutation in the adiponectin gene, in which isoleucine at position 164 in the globular domain is substituted with threonine (I164T). Subjects with this mutation showed markedly low level of plasma adiponectin and clinical features of the metabolic syndrome. Here, we examined the molecular characteristics of the mutant protein associated with a genetic cause of hypoadiponectinemia. The current study revealed (1) the mutant protein showed an oligomerization state similar to the wild-type as determined by gel filtration chromatography and, (2) the mutant protein exhibited normal insulin-sensitizing activity, but (3) pulse-chase study showed abnormal secretion of the mutant protein from adipose tissues. Our results suggest that I164T mutation is associated with hypoadiponectinemia through disturbed secretion into plasma, which may contribute to the development of the metabolic syndrome.
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Targeted Inhibition of EGFR and Glutaminase Induces Metabolic Crisis in EGFR Mutant Lung Cancer.
Momcilovic, Milica; Bailey, Sean T; Lee, Jason T; Fishbein, Michael C; Magyar, Clara; Braas, Daniel; Graeber, Thomas; Jackson, Nicholas J; Czernin, Johannes; Emberley, Ethan; Gross, Matthew; Janes, Julie; Mackinnon, Andy; Pan, Alison; Rodriguez, Mirna; Works, Melissa; Zhang, Winter; Parlati, Francesco; Demo, Susan; Garon, Edward; Krysan, Kostyantyn; Walser, Tonya C; Dubinett, Steven M; Sadeghi, Saman; Christofk, Heather R; Shackelford, David B
2017-01-17
Cancer cells exhibit increased use of nutrients, including glucose and glutamine, to support the bioenergetic and biosynthetic demands of proliferation. We tested the small-molecule inhibitor of glutaminase CB-839 in combination with erlotinib on epidermal growth factor receptor (EGFR) mutant non-small cell lung cancer (NSCLC) as a therapeutic strategy to simultaneously impair cancer glucose and glutamine utilization and thereby suppress tumor growth. Here, we show that CB-839 cooperates with erlotinib to drive energetic stress and activate the AMP-activated protein kinase (AMPK) pathway in EGFR (del19) lung tumors. Tumor cells undergo metabolic crisis and cell death, resulting in rapid tumor regression in vivo in mouse NSCLC xenografts. Consistently, positron emission tomography (PET) imaging with 18 F-fluoro-2-deoxyglucose ( 18 F-FDG) and 11 C-glutamine ( 11 C-Gln) of xenografts indicated reduced glucose and glutamine uptake in tumors following treatment with CB-839 + erlotinib. Therefore, PET imaging with 18 F-FDG and 11 C-Gln tracers can be used to non-invasively measure metabolic response to CB-839 and erlotinib combination therapy. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.
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Targeted Inhibition of EGFR and Glutaminase Induces Metabolic Crisis in EGFR Mutant Lung Cancer
Directory of Open Access Journals (Sweden)
Milica Momcilovic
2017-01-01
Full Text Available Cancer cells exhibit increased use of nutrients, including glucose and glutamine, to support the bioenergetic and biosynthetic demands of proliferation. We tested the small-molecule inhibitor of glutaminase CB-839 in combination with erlotinib on epidermal growth factor receptor (EGFR mutant non-small cell lung cancer (NSCLC as a therapeutic strategy to simultaneously impair cancer glucose and glutamine utilization and thereby suppress tumor growth. Here, we show that CB-839 cooperates with erlotinib to drive energetic stress and activate the AMP-activated protein kinase (AMPK pathway in EGFR (del19 lung tumors. Tumor cells undergo metabolic crisis and cell death, resulting in rapid tumor regression in vivo in mouse NSCLC xenografts. Consistently, positron emission tomography (PET imaging with 18F-fluoro-2-deoxyglucose (18F-FDG and 11C-glutamine (11C-Gln of xenografts indicated reduced glucose and glutamine uptake in tumors following treatment with CB-839 + erlotinib. Therefore, PET imaging with 18F-FDG and 11C-Gln tracers can be used to non-invasively measure metabolic response to CB-839 and erlotinib combination therapy.
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Material properties of the F82H melted in an electric arc furnace
Energy Technology Data Exchange (ETDEWEB)
Sakasegawa, Hideo, E-mail: sakasegawa.hideo@jaea.go.jp [Japan Atomic Energy Agency, Rokkasho, Aomori (Japan); Tanigawa, Hiroyasu [Japan Atomic Energy Agency, Rokkasho, Aomori (Japan); Kano, Sho; Abe, Hiroaki [Institute for Materials Research, Tohoku university, Sendai, Miyagi (Japan)
2015-10-15
Highlights: • We studied material properties of reduced activation ferritic/martensitic steel. • We melted F82H using a 20 tons electric arc furnace for the first time. • Mass effect likely affected material properties. • MX (M: Metal, C: Carbon and/or Nitrogen) precipitates mainly formed on grain and sub grain boundaries. - Abstract: Fusion DEMO reactor requires over 11,000 tons of reduced activation ferritic/martensitic steel. It is necessary to develop the manufacturing technology for fabricating such large-scale steel with appropriate mechanical properties. In this work, we focused fundamental mechanical properties and microstructures of F82H-BA12 heat which was melted using a 20 tons electric arc furnace followed by electroslag remelting process. Its raw material of iron was blast furnace iron, because the production volume of electrolytic iron which has been used in former heats, is limited. After melting and forging, this F82H-BA12 heat was heat-treated in four different conditions to consider their fluctuations and to optimize them, and tensile and Charpy impact tests were then performed. The result of these mechanical properties were comparable to those of former F82H heats less than 5 tons which were melted applying vacuum induction melting.
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Mahajan, Sai Pooja; Velez-Vega, Camilo; Escobedo, Fernando A
2013-01-10
Nanobodies are single-domain antibodies found in camelids. These are the smallest naturally occurring binding domains and derive functionality via three hypervariable loops (H1-H3) that form the binding surface. They are excellent candidates for antibody engineering because of their favorable characteristics like small size, high solubility, and stability. To rationally engineer antibodies with affinity for a specific target, the hypervariable loops can be tailored to obtain the desired binding surface. As a first step toward such a goal, we consider the design of loops with a desired conformation. In this study, we focus on the H1 loop of the anti-hCG llama nanobody that exhibits a noncanonical conformation. We aim to "tilt" the stability of the H1 loop structure from a noncanonical conformation to a (humanized) type 1 canonical conformation by studying the effect of selected mutations to the amino acid sequence of the H1, H2, and proximal residues. We use all-atomistic, explicit-solvent, biased molecular dynamic simulations to simulate the wild-type and mutant loops in a prefolded framework. We thus find mutants with increasing propensity to form a stable type 1 canonical conformation of the H1 loop. Free energy landscapes reveal the existence of conformational isomers of the canonical conformation that may play a role in binding different antigenic surfaces. We also elucidate the approximate mechanism and kinetics of transitions between such conformational isomers by using a Markovian model. We find that a particular three-point mutant has the strongest thermodynamic propensity to form the H1 type 1 canonical structure but also to exhibit transitions between conformational isomers, while a different, more rigid three-point mutant has the strongest propensity to be kinetically trapped in such a canonical structure.
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Influenza A virus NS1 gene mutations F103L and M106I increase replication and virulence
Directory of Open Access Journals (Sweden)
Ping Jihui
2011-01-01
Full Text Available Abstract Background To understand the evolutionary steps required for a virus to become virulent in a new host, a human influenza A virus (IAV, A/Hong Kong/1/68(H3N2 (HK-wt, was adapted to increased virulence in the mouse. Among eleven mutations selected in the NS1 gene, two mutations F103L and M106I had been previously detected in the highly virulent human H5N1 isolate, A/HK/156/97, suggesting a role for these mutations in virulence in mice and humans. Results To determine the selective advantage of these mutations, reverse genetics was used to rescue viruses containing each of the NS1 mouse adapted mutations into viruses possessing the HK-wt NS1 gene on the A/PR/8/34 genetic backbone. Both F103L and M106I NS1 mutations significantly enhanced growth in vitro (mouse and canine cells and in vivo (BALB/c mouse lungs as well as enhanced virulence in the mouse. Only the M106I NS1 mutation enhanced growth in human cells. Furthermore, these NS1 mutations enhanced early viral protein synthesis in MDCK cells and showed an increased ability to replicate in mouse interferon β (IFN-β pre-treated mouse cells relative to rPR8-HK-NS-wt NS1. The double mutant, rPR8-HK-NS-F103L + M106I, demonstrated growth attenuation late in infection due to increased IFN-β induction in mouse cells. We then generated a rPR8 virus possessing the A/HK/156/97 NS gene that possesses 103L + 106I, and then rescued the L103F + I106M mutant. The 103L + 106I mutations increased virulence by >10 fold in BALB/c mice. We also inserted the avian A/Ck/Beijing/1/95 NS1 gene (the source lineage of the A/HK/156/97 NS1 gene that possesses 103L + 106I, onto the A/WSN/33 backbone and then generated the L103F + I106M mutant. None of the H5N1 and H9N2 NS containing viruses resulted in increased IFN-β induction. The rWSN-A/Ck/Beijing/1/95-NS1 gene possessing 103L and 106I demonstrated 100 fold enhanced growth and >10 fold enhanced virulence that was associated with increased tropism for lung
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Cloning a T-DNA-Linked Phosphate Gene that mediates Salt Tolerance on Mutant of Arabidopsis thaliana
International Nuclear Information System (INIS)
Njoroge, N.C; Tremblay, L.; Lefebvre, D.D.
2006-01-01
T-DNA insertionally mutagenized seeds of Arabidopsis thaliana were used to unravel genetic mechanisms underlying salt tolerance in plants. Over a period of two weeks, kanamycin homozygous (KK) seeds of the mutant NN143 attain germination levels of 65% and 77% on 175mM Nacl and 300mM mannitol respectively. Under these conditions of osmotic stress, the wild type seeds were incapable of germination. The mutant was also capable of germination on a medium containing 2μM abscisic acid (ABA). After two weeks on 2μM ABA, it attained 100% germination and the wild type did not germinate. The ABA level in the mutant was 40% higher than the wild type. Segregation analysis indicated that salt tolerance in the mutant is T-DNA linked. Genetic analysis of the F1 and F2 generations indicated that the salt tolerance trait in the mutant is dominant. The putative salt tolerance gene of mutant NN143 was cloned by plasmid rescue and sequence data indicated involvement of a protein phosphatase. The possible mechanism underlying salt tolerance in the mutant is discussed.(author)
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Characterization of a novel curled-cotyledons mutant in soybean ...
African Journals Online (AJOL)
ARL
different stages. Protein and amino acid contents in seeds of mutant are higher than those of the wild .... curled shape, from seedling to maturation phase (Figure. 1D-1F). ..... Arabidopsis seed dermination by stimulating abscisic acid synthesis.
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International Nuclear Information System (INIS)
Kukimura, H.; Ikeda, F.; Fujita, H.; Maeta, T.; Nakajima, K.; Katagiri, K.; Nakahira, K.; Somegou, M.
1976-01-01
The plants studied included apple trees, cryptomeria (japanese cedar) and mulberry. In apple, dwarf and compact types of mutants from cv. Fuji were found to be graft incompatible on Maruba-kaido(Malus prunifolia) rootstock. In Sunki mandarin(Citrus sunki), the number of nucellar embryo per seed was affected by gamma-irradiation, and morphological mutants from nucellar seedlings were obtained at high rate by irradiation at floral bud stage with 2kR exposure. In Cryptomeria, re-irradiated waxless mutants by gamma-rays showed very high rate of somatic mutation when compared to other morphological mutants. Pollen sterility and pollen shaped PMC were found in the most of gamma-induced-mutants. Mutants forming pollen shaped PMC had a genetical tendency of continuous male flower bud formation for a longer term. With mulberry, time of sprouting of induced mutants differed from the originals. Ability of root initiation of semi-softwood cuttings in morphological mutants were tested. Cytochimera induction were found at considerably high rate when actively growing diploid plants were irradiated by gamma-rays. Eight kinds of cytochimeras were induced. Frequency of 2-4-4 was extremely high(approx. 50%), then 4-2-2 and 2-4-2 chimeras followed. Seven kinds were induced by semi-acute irradiation(200R/h), while 4 kinds by acute irradiation(5kR/h). By breeding test it was cleared that the elongate and entire leaf was sexually transmissible, whereas the 'dwarf' was not obvious and the 'marginally curledleaf' was not transmissible. Pyronin-methylgreen staining method proved to be useful in some morphological mutants to distinguish the histo-genetical differences which exist in the shoot apex.
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Semi-dwarf mutants in triticale and wheat breeding
International Nuclear Information System (INIS)
Driscoll, C.J.
1984-01-01
The triticale lines Beagle and DR-IRA have been subjected to ionizing irradiation and chemical mutagenesis in order to produce semi-dwarf mutants. Beagle is 100 cm tall and DR-IRA 80 cm under average field conditions. A bulk then pedigree method is currently represented by 158 single plots of M 6 (or in some cases M 7 ) mutants that are from 5 to 35 cm shorter than the control variety. The shortest mutants are 65 cm in height. Forty of these mutants are also earlier flowering than the control varieties. Replicated yield testing will be conducted on confirmed mutants in 1983. Response to gibberellic acid of these mutants will also be determined. The Cornerstone male-sterility mutant (ms1c) on chromosome arm 4Aα has been combined with the GA-insensitive/reduced height gene Gai/Rht1 which is also on chromosome arm 4Aα. The ms1c mutant has also been combined with Gai/Rht2 on chromosome 4D and with both Gai/Rht1 and Gai/Rht2. The combination ms1c and Gai/Rht1 has been chosen as the basis of a composite cross. Thirteen varieties were tested with GA 3 and seven (Warigal, Aroona, Oxley, Banks, Avocet, Matipo and Toquifen) which contain Gai/Rht1 were crossed with ms1c Gai/Rht1 and entered into an interpollinating F 2 . The entire composite is homozygous for this semi-dwarf allele and selection will be practiced for increased height on a GA-insensitive background. (author)
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21 CFR 184.1027 - Mixed carbohydrase and protease enzyme product.
2010-04-01
... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Mixed carbohydrase and protease enzyme product. 184... RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 184.1027 Mixed carbohydrase and protease enzyme product. (a) Mixed carbohydrase and protease enzyme product is an enzyme preparation that includes...
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Sulfhydryl oxidation of mutants with cysteine in place of acidic residues in the lactose permease.
Voss, J; Sun, J; Venkatesan, P; Kaback, H R
1998-06-02
To examine further the role of charge-pair interactions in the structure and function of lactose permease, Asp237 (helix VII), Asp240 (helix VII), Glu126 (cytoplasmic loop IV/V), Glu269 (helix VIII), and Glu325 (helix X) were replaced individually with Cys in a functional mutant devoid of Cys residues. Each mutant was then oxidized with H2O2 in order to generate a sulfinic and/or sulfonic acid at these positions. Due to the isosteric relationship between aspartate and sulfinate, in particular, and the lower pKa of the sulfinic and sulfonic acid side chains, oxidized derivatives of Cys are useful probes for examining the role of carboxylates. Asp237-->Cys or Asp240-->Cys permease is inactive, as shown previously, but H2O2 oxidation restores activity to an extent similar to that observed when a negative charge is reintroduced by other means. Glu126-->Cys, Glu269-->Cys, or Glu325-->Cys permease is inactive, but oxidation does not restore active lactose transport. The data are consistent with previous observations indicating that Asp237 and Asp240 are not critical for active lactose transport, while Glu126, Glu269, and Glu325 are irreplaceable. Although Glu269-->Cys permease does not transport lactose, the oxidized mutant exhibits significant transport of beta,D-galactosylpyranosyl 1-thio-beta,D-galactopyranoside, a property observed with Glu269-->Asp permease. The observation supports the idea that an acidic residue at position 269 is important for substrate recognition. Finally, oxidized Glu325-->Cys permease catalyzes equilibrium exchange with an apparent pKa of about 6.5, more than a pH unit lower than that observed with Glu325-->Asp permease, thereby providing strong confirmatory evidence that a negative charge at position 325 determines the rate of translocation of the ternary complex between the permease, substrate, and H+.
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GPCR Interaction: 184 [GRIPDB[Archive
Lifescience Database Archive (English)
Full Text Available tion of C5aR dimers.Upon C5a stimulation, mutant receptors did not internalize when individually express...ed. C5a stimulation of cells that co-expressed wild type C5aR together with either un...terfaces of C5aR A Complement component C5a C5aR A Complement component C5a C5aR Experiment Neither an intact NH2 term...inus nor the presence of COOH-terminal phosphorylation sites appeared to be required for the forma...liganded or phosphorylation-deficient mutant resulted in co-internalization of mutant receptors with C5aR. 18772131 NH2-terminally modified C5aR mutant NP_001727.1 ...
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International Nuclear Information System (INIS)
Kalantaryan, V.P.; Trchounian, A.H.; Soghomonyan, D.R.
2014-01-01
The aim of this study was the investigation of the effects of low intensity electromagnetic irradiation (EMI) at the frequencies of 51.8 and 53 GHz and of antibiotic ceftazidime on the N,N'-dicyclohexylcarbodiimide (DCCD) inhibited ATPase activity of membrane vesicles of lactic acid bacteria Lactobacillus acidophilus grown at low pH (pH 4.0 or 6.5) and assayed at the same pH. It was shown that both frequencies EMI stimulated ATPase activity of L. acidophilus grown at pH 4.0, but EMI combined with ceftazidime and DCCD decreased ATPase activity at pH 4.0 and pH 6.5. It was suggested that the F 0 F 1 -ATPase might be a target for EMI even at low pH
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Directory of Open Access Journals (Sweden)
Matthew Dale Woolard
2013-02-01
Full Text Available Francisella tularensis is the causative agent of tularemia. We have previously shown that infection with F. tularensis Live Vaccine Strain (LVS induces macrophages to synthesize prostaglandin E2 (PGE2. Synthesis of PGE2 by F. tularensis infected macrophages results in decreased T cell proliferation in vitro and increased bacterial survival in vivo. Although we understand some of the biological consequences of F. tularensis induced PGE2 synthesis by macrophages, we do not understand the cellular pathways (neither host nor bacterial that result in up-regulation of the PGE2 biosynthetic pathway in F. tularensis infected macrophages. We took a genetic approach to begin to understand the molecular mechanisms of bacterial induction of PGE2 synthesis from infected macrophages. To identify F. tularensis genes necessary for the induction of PGE2 in primary macrophages, we infected cells with individual mutants from the closely related strain Francisella tularensis subspecies novicida U112 (U112 two allele mutant library. Twenty genes were identified that when disrupted resulted in U112 mutant strains unable to induce the synthesis of PGE2 by infected macrophages. Fourteen of the genes identified are located within the Francisella pathogenicity island (FPI. Genes in the FPI are required for F. tularensis to escape from the phagosome and replicate in the cytosol, which might account for the failure of U112 with transposon insertions within the FPI to induce PGE2. This implies that U112 mutant strains that do not grow intracellularly would also not induce PGE2. We found that U112 clpB::Tn grows within macrophages yet fails to induce PGE2, while U112 pdpA::Tn does not grow yet does induce PGE2. We also found that U112 iglC::Tn neither grows nor induces PGE2. These findings indicate that there is dissociation between intracellular growth and the ability of F. tularensis to induce PGE2 synthesis. These mutants provide a critical entrée into the pathways used
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Hydrogen behaviour in the aged low activation martensitic steel F82H for fusion reactor applications
International Nuclear Information System (INIS)
Benamati, G.
1997-10-01
A time dependent permeation method is used to measure the permeability, diffusivity and solubility of hydrogen in the low activation martensitic steel F82H aged for 2000 h under vacuum at 773 K. The measurements cover the temperature range from 373 to 723 K which includes the onset of hydrogen trapping effects on diffusivity and solubility. The results are interpreted using a trapping model. The number of trap sites and their average energies for hydrogen in the aged F82H steel are determined. These data are compared with those obtained for deuterium in F82H steel
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Mai, Antonello; Artico, Marino; Rotili, Dante; Tarantino, Domenico; Clotet-Codina, Imma; Armand-Ugón, Mercedes; Ragno, Rino; Simeoni, Silvia; Sbardella, Gianluca; Nawrozkij, Maxim B; Samuele, Alberta; Maga, Giovanni; Esté, José A
2007-11-01
Following the disclosure of dihydro-alkoxy-, dihydro-alkylthio-, and dihydro-alkylamino-benzyl-oxopyrimidines (DABOs, S-DABOs, and NH-DABOs) as potent and selective anti-HIV-1 agents belonging to the non-nucleoside reverse transcriptase inhibitor (NNRTI) class, we report here the synthesis and biological evaluation of a novel series of DABOs bearing a N,N-disubstituted amino group or a cyclic amine at the pyrimidine-C2 position, a hydrogen atom or a small alkyl group at C5 and/or at the benzylic position, and the favorable 2,6-difluorobenzyl moiety at the C6 position (F2-N,N-DABOs). The new compounds were highly active up to the subnanomolar level against both wt HIV-1 and the Y181C mutant and at the submicromolar to nanomolar range against the K103N and Y188L mutant strains. Such derivatives were more potent than S-DABOs, NH-DABOs, and nevirapine and efavirenz were chosen as reference drugs. The higher inhibitor adaptability to the HIV-1 RT non-nucleoside binding site (NNBS) may account for the higher inhibitory effect exerted by the new molecules against the mutated RTs.
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20 CFR 220.184 - If the annuitant becomes disabled by another impairment(s).
2010-04-01
... impairment(s). 220.184 Section 220.184 Employees' Benefits RAILROAD RETIREMENT BOARD REGULATIONS UNDER THE... Activity or Medical Improvement § 220.184 If the annuitant becomes disabled by another impairment(s). If a new severe impairment(s) begins in or before the month in which the last impairment(s) ends, the Board...
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D'Amour, Pierre; Räkel, Agnès; Brossard, Jean-Hugues; Rousseau, Louise; Albert, Caroline; Cantor, Tom
2006-01-01
The quantitative evaluation of circulating PTH peaks revealed by PTH assays after HPLC separation constitutes the best way to study the behavior of PTH molecular forms, but it is also impractical. The objective of the study was to investigate the regulation of circulating PTH molecular forms by calcium through the use of PTH fragments/PTH (1-84) ratios derived from PTH assays with different specificities before and after HPLC separation of circulating PTH. CaCl2 and Na citrate were infused in eight volunteers. PTH was measured in serum and HPLC fractions at different calcium concentrations in PTH assays reacting with regions 1-2 (CA), 12-18 (T), and 65-69 (C) of the PTH structure. From hypo- to hypercalcemia, the C/CA ratio had the highest range (1.92 to 9.75; P < 0.001), and the C/T ratio had a higher range (1.69 to 6.11; P < 0.01) than the T/CA ratio (1.15 to 1.86). Human (h) PTH (1-84) represented 32.7 and 4.3% of circulating PTH in hypo- and hypercalcemic HPLC profiles, respectively. These numbers were 5 and 0.9% for amino-terminal (N)-PTH, an amino-terminal form of PTH distinct from hPTH (1-84), 7.3 and 6.8% for non-(1-84) PTH or large C-PTH fragments with a partially preserved N structure, and 54.9 and 88.1% for C-PTH fragments missing a N structure. The HPLC C-PTH fragments to hPTH (1-84) ratio had the most extensive range (1.67 to 20.58). Despite their quantitative differences, all ratios identified identical behavior of PTH fragments relative to PTH (1-84). PTH assay ratios are an adequate tool to investigate the modulation of PTH molecular forms, even if all PTH assays show some undesirable cross-reactivity with certain circulating forms of PTH.
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Proton irradiation effects on tensile and bend-fatigue properties of welded F82H specimens
Energy Technology Data Exchange (ETDEWEB)
Saito, S., E-mail: saito.shigeru@jaea.go.j [JAEA Tokai, J-PARC Center, 2-4 Shirakata-shirane, Tokai-mura, Naka-gun, Ibaraki-ken 319-1195 (Japan); Kikuchi, K.; Hamaguchi, D. [JAEA Tokai, J-PARC Center, 2-4 Shirakata-shirane, Tokai-mura, Naka-gun, Ibaraki-ken 319-1195 (Japan); Usami, K.; Ishikawa, A.; Nishino, Y.; Endo, S. [JAEA Tokai, Department of Hot Laboratories, Tokai-mura, Ibaraki-ken 319-1195 (Japan); Kawai, M. [KEK, Tsukuba-shi, Ibaraki-ken 305-0801 (Japan); Dai, Y. [PSI, Spallation Source Division, 5232 Villigen PSI (Switzerland)
2010-03-15
In several institutes, research and development for an accelerator-driven transmutation system (ADS) have been progressed. Ferritic/martensitic (FM) steels are the candidate materials for the beam window of ADS. To evaluate of the mechanical properties of the irradiated materials, the post irradiation examination (PIE) work of the SINQ (Swiss spallation neutron source) target irradiation program (STIP) specimens was carried out at JAEA. In present study, the results of PIE on FM steel F82H and its welded joint have been reported. The present irradiation conditions of the specimens were as follows: proton energy was 580 MeV. Irradiation temperatures were ranged from 130 to 380 deg. C, and displacement damage level was ranged from 5.7 to 11.8 dpa. The results of tensile tests performed at 22 deg. C indicated that the irradiation hardening occurred with increasing the displacement damage up to 10.1 dpa at 320 deg. C irradiation. At higher dose (11.8 dpa) and higher temperature (380 deg. C), irradiation hardening was observed, but degradation of ductility was relaxed in F82H welded joint. In present study, all specimens kept its ductility after irradiation and fractured in ductile manner. The results on bend-fatigue tests showed that the fatigue life (N{sub f}) of F82H base metal irradiated up to 6.3 dpa was almost the same with that of unirradiated specimens. The N{sub f} of the specimens irradiated up to 9.1 dpa was smaller than that of unirradiated specimens. Though the number of specimen was limited, the N{sub f} of F82H EB (15 mm) and EB (3.3 mm) welded joints seemed to increase after irradiation and the fracture surfaces of the specimens showed transgranular morphology. While F82H TIG welded specimens were not fractured by 10{sup 7} cycles.
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21 CFR 184.1250 - Cellulase enzyme preparation derived from Trichoderma longibrachiatum.
2010-04-01
... Trichoderma longibrachiatum. 184.1250 Section 184.1250 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT....1250 Cellulase enzyme preparation derived from Trichoderma longibrachiatum. (a) Cellulase enzyme preparation is derived from a nonpathogenic, nontoxicogenic strain of Trichoderma longibrachiatum (formerly T...
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RgpF Is Required for Maintenance of Stress Tolerance and Virulence in Streptococcus mutans.
Kovacs, C J; Faustoferri, R C; Quivey, R G
2017-12-15
Bacterial cell wall dynamics have been implicated as important determinants of cellular physiology, stress tolerance, and virulence. In Streptococcus mutans , the cell wall is composed primarily of a rhamnose-glucose polysaccharide (RGP) linked to the peptidoglycan. Despite extensive studies describing its formation and composition, the potential roles for RGP in S. mutans biology have not been well investigated. The present study characterizes the impact of RGP disruption as a result of the deletion of rgpF , the gene encoding a rhamnosyltransferase involved in the construction of the core polyrhamnose backbone of RGP. The Δ rgpF mutant strain displayed an overall reduced fitness compared to the wild type, with heightened sensitivities to various stress-inducing culture conditions and an inability to tolerate acid challenge. The loss of rgpF caused a perturbation of membrane-associated functions known to be critical for aciduricity, a hallmark of S. mutans acid tolerance. The proton gradient across the membrane was disrupted, and the Δ rgpF mutant strain was unable to induce activity of the F 1 F o ATPase in cultures grown under low-pH conditions. Further, the virulence potential of S. mutans was also drastically reduced following the deletion of rgpF The Δ rgpF mutant strain produced significantly less robust biofilms, indicating an impairment in its ability to adhere to hydroxyapatite surfaces. Additionally, the Δ rgpF mutant lost competitive fitness against oral peroxigenic streptococci, and it displayed significantly attenuated virulence in an in vivo Galleria mellonella infection model. Collectively, these results highlight a critical function of the RGP in the maintenance of overall stress tolerance and virulence traits in S. mutans IMPORTANCE The cell wall of Streptococcus mutans , the bacterium most commonly associated with tooth decay, is abundant in rhamnose-glucose polysaccharides (RGP). While these structures are antigenically distinct to S. mutans
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Production and characterization of radiation-sensitive meiotic mutants of Coprinus cinereus
International Nuclear Information System (INIS)
Zolan, M.E.; Tremel, C.J.; Pukkila, P.J.
1988-01-01
We have isolated four gamma-sensitive mutants of the basidiomycete Coprinus cinereus. When homozygous, two of these (rad 3-1 and rad 9-1) produce fruiting bodies with very few viable basidiospores, the products of meiosis in this organism. A less radiation-sensitive allele of RAD 3, rad 3-2, causes no apparent meiotic defect in homozygous strains. Quantitative measurements of oidial survival of rad 3-1;rad 9-1 double mutants compared to the single mutants indicated that rad 3-1 and rad 9-1 mutants are defective in the same DNA repair pathway. In the pew viable basidiospores that are produced by these two strains, essentially normal levels of meiotic recombination can be detected. None of the mutants exhibits increased sensitivity to UV radiation. Cytological examination of meiotic chromosomes from mutant and wild-type fruiting bodies showed that rad 3-1 homozygous strains fail to condense and pair homologous chromosomes during prophase I. Although rad 9-1 strains are successful at chromosome pairing, meiosis is usually not completed in these mutants
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2010-10-01
... MISCELLANEOUS SYSTEMS AND EQUIPMENT Cooking and Heating § 184.200 General. Cooking and heating equipment must be suitable for marine use. Equipment designed and installed in accordance with American Boat and Yacht Council (ABYC) A-3, “Galley Stoves,” and A-7, “Boat Heating Systems,” or with National Fire Protection...
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2010-04-01
... Substances Affirmed as GRAS § 184.1585 Papain. (a) Papain (CAS Reg. No. 9001-73-4) is a proteolytic enzyme derived from Carica papaya L. Crude latex containing the enzyme is collected from slashed unripe papaya... latex or by precipitation from an aqueous solution of latex. The resulting enzyme preparation may be...
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Polyvalent horse F(Ab`) 2 snake antivenom: Development of ...
African Journals Online (AJOL)
F(ab´)2 fragments were further purified by Q-Fast Flow chromatography, concentrated by molecular ultrafiltration and sterilized by filtration through 0.22 m membranes. The resulting F(ab´)2 preparations were rich in intact L and in pieces of H IgG(T) chains, as demonstrated by electrophoresis and Western blot and exhibited ...
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International Nuclear Information System (INIS)
Newman, A.P.; Ferro-Novick, S.
1987-01-01
We have adapted a [ 3 H]mannose suicide selection to identify mutations in additional genes which function in the early part of the yeast secretory pathway. Thus far this protocol has led to the identification of two new genes which are implicated in this process, as well as additional alleles of previously identified genes. The new mutants, bet1 and bet2, are temperature sensitive for growth and protein transport. Thin section analysis has revealed the accumulation of a network of endoplasmic reticulum (ER) at the restrictive temperature (37 0 C). Precursors of exported proteins that accumulate in the cell at 37 0 C are terminally core glycosylated. These observations suggest that the transport of precursors is blocked subsequent to translocation into the ER but before entry into the Golgi apparatus. The bet1 and bet2 mutants define two new complementation groups which have the same properties as previously identified ER-accumulating mutants. This and previous findings suggest that protein exit from the ER and entry into the Golgi apparatus is a complex process requiring at least 11 genes
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A yeast mutant specifically sensitive to bifunctional alkylation
International Nuclear Information System (INIS)
Ruhland, A.; Kircher, M.; Wilborn, F.; Brendel, M.
1981-01-01
A mutation that specifically confers sensitivity to bi- and tri-functional alkylating agents is presented. No or little cross-sensitivity to radiation or monofunctional agents could be detected. Sensitivity does not seem to be due to preferential alkylation of mutant DNA as parent and mutant strain exhibit the same amount of DNA alkylation and the same pattern of DNA lesions including interstrand crosslinks. The mutation is due to a defect in a nuclear gene which has been designated SNM1 (sensitive to nitrogen mustard); it may control an important step in the repair of DNA interstrand crosslinks (orig.(AJ)
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MicroRNA-184 inhibits neuroblastoma cell survival through targeting the serine/threonine kinase AKT2
Directory of Open Access Journals (Sweden)
Murphy Derek M
2010-04-01
Full Text Available Abstract Background Neuroblastoma is a paediatric cancer of the sympathetic nervous system. The single most important genetic indicator of poor clinical outcome is amplification of the MYCN transcription factor. One of many down-stream MYCN targets is miR-184, which is either directly or indirectly repressed by this transcription factor, possibly due to its pro-apoptotic effects when ectopically over-expressed in neuroblastoma cells. The purpose of this study was to elucidate the molecular mechanism by which miR-184 conveys pro-apoptotic effects. Results We demonstrate that the knock-down of endogenous miR-184 has the opposite effect of ectopic up-regulation, leading to enhanced neuroblastoma cell numbers. As a mechanism of how miR-184 causes apoptosis when over-expressed, and increased cell numbers when inhibited, we demonstrate direct targeting and degradation of AKT2, a major downstream effector of the phosphatidylinositol 3-kinase (PI3K pathway, one of the most potent pro-survival pathways in cancer. The pro-apoptotic effects of miR-184 ectopic over-expression in neuroblastoma cell lines is reproduced by siRNA inhibition of AKT2, while a positive effect on cell numbers similar to that obtained by the knock-down of endogenous miR-184 can be achieved by ectopic up-regulation of AKT2. Moreover, co-transfection of miR-184 with an AKT2 expression vector lacking the miR-184 target site in the 3'UTR rescues cells from the pro-apoptotic effects of miR-184. Conclusions MYCN contributes to tumorigenesis, in part, by repressing miR-184, leading to increased levels of AKT2, a direct target of miR-184. Thus, two important genes with positive effects on cell growth and survival, MYCN and AKT2, can be linked into a common genetic pathway through the actions of miR-184. As an inhibitor of AKT2, miR-184 could be of potential benefit in miRNA mediated therapeutics of MYCN amplified neuroblastoma and other forms of cancer.
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Shigaki, Toshiro; Barkla, Bronwyn J; Miranda-Vergara, Maria Cristina; Zhao, Jian; Pantoja, Omar; Hirschi, Kendal D
2005-08-26
In plants, yeast, and bacteria, cation/H+ exchangers (CAXs) have been shown to translocate Ca2+ and other metal ions utilizing the H+ gradient. The best characterized of these related transporters is the plant vacuolar localized CAX1. We have used site-directed mutagenesis to assess the impact of altering the seven histidine residues to alanine within Arabidopsis CAX1. The mutants were expressed in a Saccharomyces cerevisiae strain that is sensitive to Ca2+ and other metals. By utilizing a yeast growth assay, the H338A mutant was the only mutation that appeared to alter Ca2+ transport activity. The CAX1 His338 residue is conserved among various CAX transporters and may be located within a filter for cation selection. We proceeded to mutate His338 to every other amino acid residue and utilized yeast growth assays to estimate the transport properties of the 19 CAX mutants. Expression of 16 of these His338 mutants could not rescue any of the metal sensitivities. However, expression of H338N, H338Q, and H338K allowed for some growth on media containing Ca2+. Most interestingly, H338N exhibited increased tolerance to Cd2+ and Zn2+. Endomembrane fractions from yeast cells were used to measure directly the transport of H338N. Although the H338N mutant demonstrated 25% of the wild type Ca2+/H+ transport, it showed an increase in transport for both Cd2+ and Zn2+ reflected in a decrease in the Km for these substrates. This study provides insights into the CAX cation filter and novel mechanisms by which metals may be partitioned across membranes.
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Roy Choudhury, Swarup; Wang, Yuqi; Pandey, Sona
2014-07-01
Signalling pathways mediated by heterotrimeric G-proteins are common to all eukaryotes. Plants have a limited number of each of the G-protein subunits, with the most elaborate G-protein network discovered so far in soya bean (Glycine max, also known as soybean) which has four Gα, four Gβ and ten Gγ proteins. Biochemical characterization of Gα proteins from plants suggests significant variation in their properties compared with the well-characterized non-plant proteins. Furthermore, the four soya bean Gα (GmGα) proteins exhibit distinct biochemical activities among themselves, but the extent to which such biochemical differences contribute to their in vivo function is also not known. We used the yeast gpa1 mutant which displays constitutive signalling and growth arrest in the pheromone-response pathway as an in vivo model to evaluate the effect of distinct biochemical activities of GmGα proteins. We showed that specific GmGα proteins can be activated during pheromone-dependent receptor-mediated signalling in yeast and they display different strengths towards complementation of yeast gpa1 phenotypes. We also identified amino acids that are responsible for differential complementation abilities of specific Gα proteins. These data establish that specific plant Gα proteins are functional in the receptor-mediated pheromone-response pathway in yeast and that the subtle biochemical differences in their activity are physiologically relevant.
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Nuclear high-spin data for A = 174, 176 and 184
Energy Technology Data Exchange (ETDEWEB)
Junde, Huo [Jilin Univ. (China). Dept. of Physics
1996-06-01
Nuclear high-spin data are important in the frontier areas of nuclear structure physics. The information on A = 174, 176 and 184 mass chains from various reaction experiments together with their adopted high-spin levels and gamma transition properties are presented and discussed. High-spin data for A = 174, 176 and 184 mass chains were evaluated in 1995.
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International Nuclear Information System (INIS)
Wellner, R.B.; Ghosh, P.C.; Roecklein, B.; Wu, H.C.
1987-01-01
Increased [ 3 H]palmitate incorporation into specific cellular proteins has been reported to occur in Chinese hamster ovary and yeast mutant cells. In this paper we report studies concerning the relationship between N-linked oligosaccharide structure and [ 3 H]palmitate incorporation into proteins of Chinese hamster ovary (CHO) cells. We have compared the incorporation of [ 3 H]palmitate into proteins of wild-type and four different mutant CHO cell lines defective in various steps of N-linked protein glycosylation. Sodium dodecyl sulfate-gel electrophoretic analysis showed that three of the mutants exhibited increased [ 3 H]palmitate incorporation into several CHO cellular proteins (approximately 30,000-38,000 molecular weight) as compared to the wild-type cells. One of the affected mutants which accumulates the Man5Gn2Asn intermediate structure was examined in detail. In agreement with earlier reports, virtually all of the [ 3 H] palmitate-labeled proteins of both wild-type and mutant cell lines are membrane-bound. Pretreatment of the mutant cell line with tunicamycin blocked the increased [ 3 H]palmitate incorporation into the two specific proteins (both of approximately 30,000 molecular weight) observed in untreated cells; the decreased incorporation of [ 3 H]palmitate into the 30,000 molecular weight species was accompanied by a concomitant increase in the incorporation of [ 3 H]palmitate into two proteins of approximately 20,000 molecular weight. Pretreatment of wild-type cells with tunicamycin also caused increased [ 3 H]palmitate incorporation into the 20,000 molecular weight species
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Production of recombinant proteins in Escherichia coli tagged with the fusion protein CusF3H.
Vargas-Cortez, Teresa; Morones-Ramirez, Jose Ruben; Balderas-Renteria, Isaias; Zarate, Xristo
2017-04-01
Recombinant protein expression in the bacterium Escherichia coli still is the number one choice for large-scale protein production. Nevertheless, many complications can arise using this microorganism, such as low yields, the formation of inclusion bodies, and the requirement for difficult purification steps. Most of these problems can be solved with the use of fusion proteins. Here, the use of the metal-binding protein CusF3H+ is described as a new fusion protein for recombinant protein expression and purification in E. coli. We have previously shown that CusF produces large amounts of soluble protein, with low levels of formation of inclusion bodies, and that proteins can be purified using IMAC resins charged with Cu(II) ions. CusF3H+ is an enhanced variant of CusF, formed by the addition of three histidine residues at the N-terminus. These residues then can bind Ni(II) ions allowing improved purity after affinity chromatography. Expression and purification of Green Fluorescent Protein tagged with CusF3H+ showed that the mutation did not alter the capacity of the fusion protein to increase protein expression, and purity improved considerably after affinity chromatography with immobilized nickel ions; high yields are obtained after tag-removal since CusF3H+ is a small protein of just 10 kDa. Furthermore, the results of experiments involving expression of tagged proteins having medium to large molecular weights indicate that the presence of the CusF3H+ tag improves protein solubility, as compared to a His-tag. We therefore endorse CusF3H+ as a useful alternative fusion protein/affinity tag for production of recombinant proteins in E. coli. Copyright © 2017 Elsevier Inc. All rights reserved.
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Biodegradation of dioxins by recombinant Escherichia coli expressing rat CYP1A1 or its mutant
Energy Technology Data Exchange (ETDEWEB)
Shinkyo, Raku; Inouye, Kuniyo [Kyoto Univ. (Japan). Div. of Food Science and Biotechnology; Kamakura, Masaki; Ikushiro, Shin-ichi; Sakaki, Toshiyuki [Toyama Prefectural Univ. (Japan). Biotechnology Research Center
2006-09-15
Among polychlorinated dibenzo-p-dioxins (PCDDs), 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TetraCDD) is the most toxic one. Recently, we reported that rat CYP1A1 mutant, F240A, expressed in yeast showed metabolic activity toward 2,3,7,8-TetraCDD. In this study, we successfully expressed N-terminal truncated P450s ({delta}1A1 and {delta}F240A) in Escherichia coli cells. Kinetic analysis using membrane fractions prepared from the recombinant E. coli cells revealed that {delta}F240A has enzymatic properties similar to F240A expressed in yeast. The metabolism of PCDDs by recombinant E. coli cells expressing both {delta}F240A and human NADPH-P450 reductase was also examined. When 2,3,7-TriCDD was added to the E. coli cell culture at a final concentration of 10 {mu}M, approximately 90% of the 2,3,7-TriCDD was converted into multiple metabolites within 8 h. These results indicate the possible application of prokaryotic cells expressing {delta}F240A to the bioremediation of PCDD-contaminated soil. (orig.)
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International Nuclear Information System (INIS)
Madiraju, M.V.; Templin, A.; Clark, A.J.
1988-01-01
A mutation partially suppressing the UV sensitivity caused by recF143 in a uvrA6 background was located at codon 37 of recA where GTG (valine) became ATG (methionine). This mutation, originally named srf-803, was renamed recA803. Little if any suppression of the recF143 defect in UV induction of a lexA regulon promoter was detected. This led to the hypothesis that a defect in recombination repair of UV damage was suppressed by recA803. The mutant RecA protein (RecA803) was purified and compared with wild-type protein (RecA+) as a catalyst of formation of joint molecules. Under suboptimal conditions, RecA803 produces both a higher rate of formation and a higher yield of joint molecules. The suboptimal conditions tested included addition of single-stranded DNA binding protein to single-stranded DNA prior to addition of RecA. We hypothesize that the ability of RecA803 to overcome interference by single-stranded DNA binding protein is the property that allows recA803 to suppress partially the deficiency in repair caused by recF mutations in the uvrA6 background. Implications of this hypothesis for the function of RecF protein in recombination are discussed
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F4+ enterotoxigenic Escherichia coli (ETEC) adhesion mediated by the major fimbrial subunit FaeG.
Xia, Pengpeng; Song, Yujie; Zou, Yajie; Yang, Ying; Zhu, Guoqiang
2015-09-01
The FaeG subunit is the major constituent of F4(+) fimbriae, associated with glycoprotein and/or glycolipid receptor recognition and majorly contributes to the pathogen attachment to the host cells. To investigate the key factor involved in the fimbrial binding of F4(+) Escherichia coli, both the recombinant E. coli SE5000 strains carrying the fae operon gene clusters that express the different types of fimbriae in vitro, named as rF4ab, rF4ac, and rF4ad, respectively, corresponding to the fimbrial types F4ab, F4ac, and F4ad, and the three isogenic in-frame faeG gene deletion mutants were constructed. The adhesion assays and adhesion inhibition assays showed that ΔfaeG mutants had a significant reduction in the binding to porcine brush border as well as the intestinal epithelial cell lines, while the complemented strain ΔfaeG/pfaeG restored the adhesion function. The recombinant bacterial strains rF4ab, rF4ac, and rF4ad have the same binding property as wild-type F4(+) E. coli strains do and improvement in terms of binding to porcine brush border and the intestinal epithelial cells, and the adherence was blocked by the monoclonal antibody anti-F4 fimbriae. These data demonstrate that the fimbrial binding of F4(+) E. coli is directly mediated by the major FaeG subunit. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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2010-08-25
... Industries GmbH Models DA 40 and DA 40F Airplanes AGENCY: Federal Aviation Administration (FAA), Department... new airworthiness directive (AD) for all Diamond Aircraft Industries GmbH Models DA 40 and DA 40F... received information from Diamond Aircraft Industries GmbH that the Models DA 40 and DA 40F airplanes have...
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Weldability of dissimilar joint between F82H and SUS316L under fiber laser welding
Energy Technology Data Exchange (ETDEWEB)
Serizawa, Hisashi [Joining and Welding Research Institute, Osaka University, 11-1 Mihogaoka, Ibaraki, Osaka 567-0047 (Japan); Mori, Daiki; Shirai, Yuma; Ogiwara, Hiroyuki; Mori, Hiroaki [Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871 (Japan)
2013-10-15
Highlights: • The microstructure of F82H/SUS316L dissimilar joint can be divided into four regions. • In the case without beam position shift, hardness of WM cannot be reduced by PWHT. • The fiber laser welding would be applicable for constructing the dissimilar joint. -- Abstract: As one of the high beam quality heat sources, 4 kW fiber laser was applied for joining between reduced activation ferritic/martensitic steel, F82H and SUS316L austenitic stainless steel, and the microstructural analyses and Vickers hardness measurements were carried out before and after post-weld heat treatment (PWHT). The microstructure of joint can be divided into four regions which are base metal of F82H, heat affected zone (HAZ) in F82H, weld metal (WM) and base metal of SUS316L. Also, it is revealed that the high-power fiber laser can be employed for constructing butt joint between F82H and SUS316L by applying PWHT and shifting the laser beam position to SUS316L, where the distance between the contact face and beam should be set as a range from radius to diameter of laser beam.
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Weldability of dissimilar joint between F82H and SUS316L under fiber laser welding
International Nuclear Information System (INIS)
Serizawa, Hisashi; Mori, Daiki; Shirai, Yuma; Ogiwara, Hiroyuki; Mori, Hiroaki
2013-01-01
Highlights: • The microstructure of F82H/SUS316L dissimilar joint can be divided into four regions. • In the case without beam position shift, hardness of WM cannot be reduced by PWHT. • The fiber laser welding would be applicable for constructing the dissimilar joint. -- Abstract: As one of the high beam quality heat sources, 4 kW fiber laser was applied for joining between reduced activation ferritic/martensitic steel, F82H and SUS316L austenitic stainless steel, and the microstructural analyses and Vickers hardness measurements were carried out before and after post-weld heat treatment (PWHT). The microstructure of joint can be divided into four regions which are base metal of F82H, heat affected zone (HAZ) in F82H, weld metal (WM) and base metal of SUS316L. Also, it is revealed that the high-power fiber laser can be employed for constructing butt joint between F82H and SUS316L by applying PWHT and shifting the laser beam position to SUS316L, where the distance between the contact face and beam should be set as a range from radius to diameter of laser beam
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Mutant breeding of Aspergillus niger irradiated by 12C6+ for hyper citric acid
International Nuclear Information System (INIS)
Hu Wei; Li Wenjian; Chen Jihong; Liu Jing; Wang Shuyang; Wang Jufang; Lu Dong
2014-01-01
In this study, strains of Aspergillus niger No.4 for hyper citric acid were irradiated to different doses by 80 MeV/u 12 C 6+ ion beams. Seven mutant strains showed marked citric acid over-production records and faster productivity than initial Aspergillus niger No.4 by shaking flash fermentation. The maximum product yield was 132.8 gL -1 (the H4002 strain) being a 8.8% increase to the initial strain. The scale-up experiment was carried out in a 100 L bioreactor. The mutant H4002 can accumulate 187gL -1 product yield of citric acid from starch liquefying supernatant. The productivity of citric acid was 2.75 g L -1 h -1 . So, the mutant H4002 possesses rapid sugar katabolism for producing citric acid. Meanwhile, the pellet morphology kept compact and round during the whole submerged fermentation, which was suited to produce citric acid. The results indicate that mutant H4002 has potential ability to produce citric acid rapidly. (authors)
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RPA binds histone H3-H4 and functions in DNA replication-coupled nucleosome assembly.
Liu, Shaofeng; Xu, Zhiyun; Leng, He; Zheng, Pu; Yang, Jiayi; Chen, Kaifu; Feng, Jianxun; Li, Qing
2017-01-27
DNA replication-coupled nucleosome assembly is essential to maintain genome integrity and retain epigenetic information. Multiple involved histone chaperones have been identified, but how nucleosome assembly is coupled to DNA replication remains elusive. Here we show that replication protein A (RPA), an essential replisome component that binds single-stranded DNA, has a role in replication-coupled nucleosome assembly. RPA directly binds free H3-H4. Assays using a synthetic sequence that mimics freshly unwound single-stranded DNA at replication fork showed that RPA promotes DNA-(H3-H4) complex formation immediately adjacent to double-stranded DNA. Further, an RPA mutant defective in H3-H4 binding exhibited attenuated nucleosome assembly on nascent chromatin. Thus, we propose that RPA functions as a platform for targeting histone deposition to replication fork, through which RPA couples nucleosome assembly with ongoing DNA replication. Copyright © 2017, American Association for the Advancement of Science.
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Directory of Open Access Journals (Sweden)
Céline Pesce
2017-06-01
Full Text Available Xanthomonas translucens is the causal agent of bacterial leaf streak, the most common bacterial disease of wheat and barley. To cause disease, most xanthomonads depend on a highly conserved type III secretion system, which translocates type III effectors into host plant cells. Mutagenesis of the conserved type III secretion gene hrcT confirmed that the X. translucens type III secretion system is required to cause disease on the host plant barley and to trigger a non-host hypersensitive response (HR in pepper leaves. Type III effectors are delivered to the host cell by a surface appendage, the Hrp pilus, and a translocon protein complex that inserts into the plant cell plasma membrane. Homologs of the Xanthomonas HrpF protein, including PopF from Ralstonia solanacearum and NolX from rhizobia, are thought to act as a translocon protein. Comparative genomics revealed that X. translucens strains harbor a noncanonical hrp gene cluster, which rather shares features with type III secretion systems from Ralstonia solanacearum, Paraburkholderia andropogonis, Collimonas fungivorans, and Uliginosibacterium gangwonense than other Xanthomonas spp. Surprisingly, none of these bacteria, except R. solanacearum, encode a homolog of the HrpF translocon. Here, we aimed at identifying a candidate translocon from X. translucens. Notably, genomes from strains that lacked hrpF/popF/nolX instead encode another gene, called hpaT, adjacent to and co-regulated with the type III secretion system gene cluster. An insertional mutant in the X. translucens hpaT gene, which is the first gene of a two-gene operon, hpaT-hpaH, was non-pathogenic on barley and did not cause the HR or programmed cell death in non-host pepper similar to the hrcT mutant. The hpaT mutant phenotypes were partially complemented by either hpaT or the downstream gene, hpaH, which has been described as a facilitator of translocation in Xanthomonas oryzae. Interestingly, the hpaT mutant was also complemented
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International Nuclear Information System (INIS)
Ogawa, T.; Kojima, I.; Takeda, N.; Fukuda, H.
1994-01-01
A nicotinic acid-resistant mutant, designated NA201, was obtained from Fusarium sp. no.5-128B by treatment with ultraviolet light. This mutant strain could grow in the presence of up to 500mM nicotinic acid in the culture medium, although the parent strain could not grow at concentrations of nicotinic acid above 200 mM. The Na201 strain exhibited morphological mutations, neither forming aerial hyphae nor secreting a red-brown pigment. However, it retained the resistance to kabicidin at 25 mg-l(-1) of the parent strain. The mutant NA201 cells contained high levels of squalene and low levels of ergosterol, about 53 times higher and five to six times lower, respectively, than those of the parent strain under standard culture conditions. The volumetric oxygen transfer coefficient (Kd) affected the level of squalene in the mutant cells. The Kd for the maximum production of squalene by the mutant was 24 mmol O2 l(-1)h(-1)atm(-1) and the level of squalene in the mutant cells was 26 mg (g cell)(-1) on a dry weight basis. The greatest accumulation of squalene by the Na201 strain, corresponding to 323 mg per liter of culture medium and 35 mg (g cell)(-1) on a dry weight basis, was achieved in a culture in which the Kd was changed from a high to a low value on the third day, with the simultaneous addition of 3% glucose (w/v)
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Screening of a lactobacillus plantarum mutant with high cla productivity induced by n+ implantation
International Nuclear Information System (INIS)
Li Shurong; Meng Xianjun; Zhang Tao; Zhao Hongwei; Lu Jiaping; Zhao Yun; Gao Yanhong; Li Qingpeng
2009-01-01
The initial lactic acid bacteria strain A6-1 was treated by N + ions implantation of 50 keV with doses of 1 x 10 13 , 3 x 10 13 , 5 x 10 13 , 8 x 10 13 , 10 x 10 13 , 30 x 10 13 , 50 x 10 13 , 80 x 10 13 , and 100 x 10 13 ions/cm 2 . The survival curve showed a saddle model, and the high survival rate was 20% ∼ 35% from the treatments of 30 x 10 13 ions/cm 2 and 50 x 10 13 ions/cm 2 implantation. Considering the survival rate, positive mutation and range of mutation rate, N + ions implantation of 30 x 10 13 ions/cm 2 was recommended for mutation breeding of lactic acid bacteria. Selected mutants with high ability of producing CLA after fermentation. Generic stable was observed until 8 generations of F mutant, and average yield of CLA was 162.5 μg/ml, which was 69.87% higher than the original stain. F mutant was named A6-1F. (authors)
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Thermoluminescence characterization of LiMgF3: DyF3 phosphors exposed to beta radiation
International Nuclear Information System (INIS)
Bernal, R.; Alday S, K.R.; Brown, F.; Cruz V, C.; Furetta, C.; Cruz Z, E.; Kitis, G.
2006-01-01
LiMgF 3 : DyF 3 phosphors were obtained as polycrystalline solids from the melting of component salts, with two DyF 3 concentrations, 2.02 % mol, and 4.04 % mol. In order to guarantee the homogeneity of composition, the samples were crushed and the resulting powder was pressed to form pellet-shaped phosphors, some of which were sintered at 700 C for 5 h under air atmosphere. Thermoluminescence measurements of beta irradiated samples show that these phosphors exhibit adequate properties to be considered for development of thermoluminescence dosimeters. Integrated thermoluminescence as a function of dose displays a linear dependence with dose for doses below 20.0 Gy. (Author)
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Role of the DELSEED Loop in Torque Transmission of F1-ATPase
Tanigawara, Mizue; Tabata, Kazuhito V.; Ito, Yuko; Ito, Jotaro; Watanabe, Rikiya; Ueno, Hiroshi; Ikeguchi, Mitsunori; Noji, Hiroyuki
2012-01-01
F1-ATPase is an ATP-driven rotary motor that generates torque at the interface between the catalytic β-subunits and the rotor γ-subunit. The β-subunit inwardly rotates the C-terminal domain upon nucleotide binding/dissociation; hence, the region of the C-terminal domain that is in direct contact with γ—termed the DELSEED loop—is thought to play a critical role in torque transmission. We substituted all the DELSEED loop residues with alanine to diminish specific DELSEED loop-γ interactions and with glycine to disrupt the loop structure. All the mutants rotated unidirectionally with kinetic parameters comparable to those of the wild-type F1, suggesting that the specific interactions between DELSEED loop and γ is not involved in cooperative interplays between the catalytic β-subunits. Glycine substitution mutants generated half the torque of the wild-type F1, whereas the alanine mutant generated comparable torque. Fluctuation analyses of the glycine/alanine mutants revealed that the γ-subunit was less tightly held in the α3β3-stator ring of the glycine mutant than in the wild-type F1 and the alanine mutant. Molecular dynamics simulation showed that the DELSEED loop was disordered by the glycine substitution, whereas it formed an α-helix in the alanine mutant. Our results emphasize the importance of loop rigidity for efficient torque transmissions. PMID:23009846
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Pudupakam, R S; Huang, Y W; Opriessnig, T; Halbur, P G; Pierson, F W; Meng, X J
2009-01-01
Hepatitis E virus (HEV) is an important human pathogen, although little is known about its biology and replication. Comparative sequence analysis revealed a hypervariable region (HVR) with extensive sequence variations in open reading frame 1 of HEV. To elucidate the role of the HVR in HEV replication, we first constructed two HVR deletion mutants, hHVRd1 and hHVRd2, with in-frame deletion of amino acids (aa) 711 to 777 and 747 to 761 in the HVR of a genotype 1 human HEV replicon. Evidence of HEV replication was detected in Huh7 cells transfected with RNA transcripts from mutant hHVRd2, as evidenced by expression of enhanced green fluorescent protein. To confirm the in vitro results, we constructed three avian HEV mutants with various HVR deletions: mutants aHVRd1, with deletion of aa 557 to 585 (Delta557-585); aHVRd2 (Delta612-641); and aHVRd3 (Delta557-641). Chickens intrahepatically inoculated with capped RNA transcripts from mutants aHVRd1 and aHVRd2 developed active viral infection, as evidenced by seroconversion, viremia, and fecal virus shedding, although mutant aHVRd3, with complete HVR deletion, was apparently attenuated in chickens. To further verify the results, we constructed four additional HVR deletion mutants using the genotype 3 swine HEV as the backbone. Mutants sHVRd2 (Delta722-781), sHVRd3 (Delta735-765), and sHVRd4 (Delta712-765) were shown to tolerate deletions and were infectious in pigs intrahepatically inoculated with capped RNA transcripts from the mutants, whereas mutant sHVRd1 (Delta712-790), with a nearly complete HVR deletion, exhibited an attenuation phenotype in infected pigs. The data from these studies indicate that deletions in HVR do not abolish HEV infectivity in vitro or in vivo, although evidence for attenuation was observed for HEV mutants with a larger or nearly complete HVR deletion.
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Functional rescue of mutant ABCA1 proteins by sodium 4-phenylbutyrate.
Sorrenson, Brie; Suetani, Rachel J; Williams, Michael J A; Bickley, Vivienne M; George, Peter M; Jones, Gregory T; McCormick, Sally P A
2013-01-01
Mutations in the ATP-binding cassette transporter A1 (ABCA1) are a major cause of decreased HDL cholesterol (HDL-C), which infers an increased risk of cardiovascular disease (CVD). Many ABCA1 mutants show impaired localization to the plasma membrane. The aim of this study was to investigate whether the chemical chaperone, sodium 4-phenylbutyrate (4-PBA) could improve cellular localization and function of ABCA1 mutants. Nine different ABCA1 mutants (p.A594T, p.I659V, p.R1068H, p.T1512M, p.Y1767D, p.N1800H, p.R2004K, p.A2028V, p.Q2239N) expressed in HEK293 cells, displaying different degrees of mislocalization to the plasma membrane and discrete impacts on cholesterol efflux, were subject to treatment with 4-PBA. Treatment restored localization to the plasma membrane and increased cholesterol efflux function for the majority of mutants. Treatment with 4-PBA also increased ABCA1 protein expression in all transfected cell lines. In fibroblast cells obtained from low HDL-C subjects expressing two of the ABCA1 mutants (p.R1068H and p.N1800H), 4-PBA increased cholesterol efflux without any increase in ABCA1 expression. Our study is the first to investigate the effect of the chemical chaperone, 4-PBA on ABCA1 and shows that it is capable of restoring plasma membrane localization and enhancing the cholesterol efflux function of mutant ABCA1s both in vitro and ex vivo. These results suggest 4-PBA may warrant further investigation as a potential therapy for increasing cholesterol efflux and HDL-C levels.
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Energy Technology Data Exchange (ETDEWEB)
Hansen, S.G.
1980-09-01
Interactions of the ions N/sup +/, F/sup +/, and CO/sub 2//sup +/ with H/sub 2/ and/or its isotopes were examined using the crossed-beam technique in the low (< 4 eV) initial relative energy. For the reaction N/sup +/(/sup 3/P) + H/sub 2/ ..-->.. NH/sup +/ + H, complex formation dominates up to 1.9 eV and a substantial interaction occurs between all collision partners up to 3.6 eV. The distribution of N/sup +/ scattered nonreactively from H/sub 2/ also showed a long-lived complex channel below 1.9 eV. The reaction F/sup +/(/sup 3/P) + H/sub 2/ ..-->..FH/sup +/ + H proceeded by a direct reaction mechanism at 0.20 to 1.07 eV. The reaction CO/sub 2//sup +/ + D/sub 2/ ..-->.. DCO/sub 2//sup +/ + D gives asymmetric product distributions at 0.27 eV and above, indicating a direct reaction mechanism. Results indicated that there are probably barriers in the exit channels for DCO/sub 2//sup +/, DCO/sup +/, and D/sub 2/O/sup +/ products. The electronic state distributions of the N/sup +/, F/sup +/, and CO/sub 2//sup +/ beams was investigated using beam attenuation and total luminescence techniques.
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Sangha, Jatinder Singh; Chen, Yolanda H; Palchamy, Kadirvel; Jahn, Gary C; Maheswaran, M; Adalla, Candida B; Leung, Hei
2008-04-01
Varietal mutants can be useful for developing durable resistance, understanding categories of resistance, and identifying candidate genes involved in defense responses. We used mutants of rice 'IR64' to isolate new sources of resistance to the planthopper Nilaparvata lugens (Stål) (Hemiptera: Delphacidae). We compared two mutants that showed a gain and loss of resistance to N. lugens, to determine the categories of resistance to this pest. Under choice tests, female planthoppers avoided settling and laid fewer eggs on the resistant mutant 'D518' than on the susceptible mutant D1131, susceptible check 'TN1', and wild-type IR64, indicating that antixenosis was the resistance category. Similarly, under no-choice conditions, planthoppers laid 29% fewer eggs in D518 than in IR64, but they oviposited more in 'D1131' and TN1. Honeydew excretion was greater on D1131 seedlings but slightly lower on D518 than on IR64. Nymphal survival and adult female weight did not differ among rice cultivars. D518 showed higher tolerance of N. lugens infestations than IR64. Genetic analysis of the F1, F2, and F3 populations derived from D518 x IR64 revealed that resistance in D518 is dominant and controlled by a single gene. Despite the variation in resistance to N. lugens, both mutants and IR64 performed similarly in the field. The mutant D518 is a new source of durable resistance to N. lugens, mainly due to enhanced antixenosis to female hoppers for settling and oviposition.
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H2A/K pseudogene mutation may promote cell proliferation
Energy Technology Data Exchange (ETDEWEB)
Guo, Jisheng; Jing, Ruirui; Lv, Xin; Wang, Xiaoyue; Li, Junqiang; Li, Lin; Li, Cuiling; Wang, Daoguang; Bi, Baibing; Chen, Xinjun [Cancer Research Center, Shandong University School of Medicine, Jinan 250012 (China); Yang, Jing-Hua, E-mail: sdu_crc_group1@126.com [Cancer Research Center, Shandong University School of Medicine, Jinan 250012 (China); Department of Surgery, VA Boston Healthcare System, Boston University School of Medicine, Boston 510660, MA (United States)
2016-05-15
Highlights: • The mutant H2A/K pseudogene is active. • The mutant H2A/K pseudogene can promote cell proliferation. - Abstract: Little attention has been paid to the histone H2A/K pseudogene. Results from our laboratory showed that 7 of 10 kidney cancer patients carried a mutant H2A/K pseudogene; therefore, we were interested in determining the relationship between mutant H2A/K and cell proliferation. We used shotgun and label-free proteomics methods to study whether mutant H2A/K lncRNAs affected cell proliferation. Quantitative proteomic analysis indicated that the expression of mutant H2A/K lncRNAs resulted in the upregulation of many oncogenes, which promoted cell proliferation. Further interaction analyses revealed that a proliferating cell nuclear antigen (PCNA)-protein interaction network, with PCNA in the center, contributes to cell proliferation in cells expressing the mutant H2A/K lncRNAs. Western blotting confirmed the critical upregulation of PCNA by mutant H2A/K lncRNA expression. Finally, the promotion of cell proliferation by mutant H2A/K lncRNAs (C290T, C228A and A45G) was confirmed using cell proliferation assays. Although we did not determine the exact mechanism by which the oncogenes were upregulated by the mutant H2A/K lncRNAs, we confirmed that the mutant H2A/K lncRNAs promoted cell proliferation by upregulating PCNA and other oncogenes. The hypothesis that cell proliferation is promoted by the mutant H2A/K lncRNAs was supported by the protein expression and cell proliferation assay results. Therefore, mutant H2A/K lncRNAs may be a new factor in renal carcinogenesis.
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Characterization of two cation diffusion facilitators NpunF0707 and NpunF1794 in Nostoc punctiforme.
Hudek, L; Pearson, L; Michalczyk, A A; Bräu, L; Neilan, B A; Ackland, M L
2015-11-01
To characterize genes involved in maintaining homeostatic levels of zinc in the cyanobacterium Nostoc punctiforme. Metal efflux transporters play a central role in maintaining homeostatic levels of trace elements such as zinc. Sequence analyses of the N. punctiforme genome identified two potential cation diffusion facilitator (CDF) metal efflux transporters, Npun_F0707 (Cdf31) and Npun_F1794 (Cdf33). Deletion of either Cdf31or Cdf33 resulted in increased zinc retention over 3 h. Interestingly, Cdf31(-) and Cdf33(-) mutants showed no change in sensitivity to zinc exposure in comparison with the wild type, suggesting some compensatory capacity for the loss of each other. Using qRT-PCR, a possible interaction was observed between the two cdf's, where the Cdf31(-) mutant had a more profound effect on cdf33 expression than Cdf33(-) did on cdf31. Over-expression of Cdf31 and Cdf33 in ZntA(-) - and ZitB(-) -deficient Escherichia coli revealed function similarities between the ZntA and ZitB of E. coli and the cyanobacterial transporters. The data presented shed light on the function of two important transporters that regulate zinc homeostasis in N. punctiforme. This study shows for the first time the functional characterization of two cyanobacterial zinc efflux proteins belonging to the CDF family. © 2015 The Society for Applied Microbiology.
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Directory of Open Access Journals (Sweden)
H. Vijay Kumar
2009-01-01
Full Text Available A method for the synthesis of tyrosine, phenyl alanine, hydroxy proline and threonine free amino acid analogues of 5H-dibenz[b,f]azepine is proposed. 5H-dibenz[b,f]azepine was prepared by known method. The key intermediate 3-chloro-1-(5H-dibenz[b,f]azepine-5-ylpropan-1-one was obtained by N-acylation of 5H-dibenz[b,f]azepine with 3-chloro propionyl chloride. Further coupling of respective free amino acid to produce 2-(3-(5H-dibenz[b,f]azepine-5-yl-3-oxopropylamino3-(4 hydroxyphenyl propanoic acid, 2-(3-(5H-dibenz[b,f]azepine-5-yl-3-oxopropylamino-3-phenyl propanoicacid,1-(3-(5H-dibenz[b,f]azepine-5-yl-3-oxopropyl-3-hydroxypyrolidine-2-carboxylic acid and 2-(3-(5H-dibenz[b,f] azepine-yl-3-oxopropyl amino-3-hydroxy butanoic acid. The synthesized compounds were evaluated for their potential over 1,1-diphenyl-2-picryl hydrazyl (DPPH free radical scavenging activity. Butylated hydroxy anisole (BHA and ascorbic acid (AA were used as the reference antioxidant compounds and also the comparative study with synthesized compounds was done. Under our experimental conditions tyrosine, hydroxy proline and threonine analogues possess a direct scavenging effect on trapping the stable free radical DPPH. Hydroxy proline analogues showed a significant radical scavenging activity among the synthesized analogues
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Zhong, Huailing; Hansen, Kasper B; Boyle, Noel J; Han, Kiho; Muske, Galina; Huang, Xinyan; Egebjerg, Jan; Sánchez, Connie
2009-10-25
The human serotonin transporter (hSERT) has primary and allosteric binding sites for escitalopram and R-citalopram. Previous studies have established that the interaction of these two compounds at a low affinity allosteric binding site of hSERT can affect the dissociation of [(3)H]escitalopram from hSERT. The allosteric binding site involves a series of residues in the 10th, 11th, and 12th trans-membrane domains of hSERT. The low affinity allosteric activities of escitalopram and R-citalopram are essentially eliminated in a mutant hSERT with changes in some of these residues, namely A505V, L506F, I507L, S574T, I575T, as measured in dissociation binding studies. We confirm that in association binding experiments, R-citalopram at clinically relevant concentrations reduces the association rate of [(3)H]escitalopram as a ligand to wild type hSERT. We demonstrate that the ability of R-citalopram to reduce the association rate of escitalopram is also abolished in the mutant hSERT (A505V, L506F, I507L, S574T, I575T), along with the expected disruption the low affinity allosteric function on dissociation binding. This suggests that the allosteric binding site mediates both the low affinity and higher affinity interactions between R-citalopram, escitalopram, and hSERT. Our data add an additional structural basis for the different efficacies of escitalopram compared to racemic citalopram reported in animal studies and clinical trials, and substantiate the hypothesis that hSERT has complex allosteric mechanisms underlying the unexplained in vivo activities of its inhibitors.
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Conserved role of unc-79 in ethanol responses in lightweight mutant mice.
Directory of Open Access Journals (Sweden)
David J Speca
2010-08-01
Full Text Available The mechanisms by which ethanol and inhaled anesthetics influence the nervous system are poorly understood. Here we describe the positional cloning and characterization of a new mouse mutation isolated in an N-ethyl-N-nitrosourea (ENU forward mutagenesis screen for animals with enhanced locomotor activity. This allele, Lightweight (Lwt, disrupts the homolog of the Caenorhabditis elegans (C. elegans unc-79 gene. While Lwt/Lwt homozygotes are perinatal lethal, Lightweight heterozygotes are dramatically hypersensitive to acute ethanol exposure. Experiments in C. elegans demonstrate a conserved hypersensitivity to ethanol in unc-79 mutants and extend this observation to the related unc-80 mutant and nca-1;nca-2 double mutants. Lightweight heterozygotes also exhibit an altered response to the anesthetic isoflurane, reminiscent of unc-79 invertebrate mutant phenotypes. Consistent with our initial mapping results, Lightweight heterozygotes are mildly hyperactive when exposed to a novel environment and are smaller than wild-type animals. In addition, Lightweight heterozygotes exhibit increased food consumption yet have a leaner body composition. Interestingly, Lightweight heterozygotes voluntarily consume more ethanol than wild-type littermates. The acute hypersensitivity to and increased voluntary consumption of ethanol observed in Lightweight heterozygous mice in combination with the observed hypersensitivity to ethanol in C. elegans unc-79, unc-80, and nca-1;nca-2 double mutants suggests a novel conserved pathway that might influence alcohol-related behaviors in humans.
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Ali, Sikander; Nawaz, Wajeeha
2017-02-01
The optimisation of nutritional requirements for dopamine (DA) synthesis by calcium alginate-entrapped mutant variant of Aspergillus oryzae EMS-6 using submerged fermentation technique was investigated. A total of 13 strains were isolated from soil. Isolate I-2 was selected as a better producer of DA and improved by exposing with ethyl methylsulphonate (EMS). EMS-6 was selected as it exhibited 43 μg/mL DA activity. The mutant variable was further treated with low levels of l-cysteine HCl to make it resistant against diversion and environmental stress. The conidiospores of mutant variant were entrapped in calcium alginate beads for stable product formation. EMS-6 gave maximum DA activity (124 μg/mL) when supplemented with 0.1% peptone and 0.2% sucrose, under optimised parameters viz. pH 3, temperature of 55 °C and incubation time of 70 min. The study involves the high profile of DA activity and is needed, as DA is capable to control numerous neurogenic disorders.
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International Nuclear Information System (INIS)
Hou Guangyun; Jing Liling
1996-01-01
A complete delia cross (no reciprocal hybrid) was made using 5 wheat mutants with distinctive characters. The heterosis and combining ability of 8 agronomic characters were studied in 10 crosses F 1 and 5 parents. The dominance degree of F 1 indicated that except for the remarkable negative heterosis for heading-date, the positive heterosis was remarkable for all the other 7 characters. The performance of parents was significantly related to the gca effects. However, gca effects were not completely consistent with the value of heterosis. As germplasm resources, dwarf-stalk mutant 890376 was the best for reducing plant height. A large grain mutant, Hesheng 2, was the best for increasing 1000-grain weight. An early maturity mutant 890236 was the best for earlier heading and fillering ability. A large head mutant 890018 was the best for improving head length, number of spikelets per head, number of grain per head and grain weight per head. To sum up, the above 4 distinctive mutants were very useful for improving the characters corresponding with their mutated characters. For some characters, the cross will have a bigger sca effects if its parents have a bigger gca effects. However, it is not the same for all their characters. Sca effects of crosses were significantly related to the heterosis over mean parent
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Uvm mutants of Escherichia coli K 12 deficient in UV mutagenesis. Pt. 1
International Nuclear Information System (INIS)
Steinborn, G.
1978-01-01
Selection for defective reversion induction, after UV treatment of E. coli K 12, yielded uvm mutants. These mutants exhibited highly reduced or no UV mutability for all loci tested although they were moderately and normally mutable by X-rays and EMS, respectively. Uvm mutations confer only a slight sensitivity to killing by UV and X-rays and no clear sensitivity to the lethal effect of HN2, EMS or MMS. Growth and viability of untreated uvm cells were normal. The properties of uvm mutants are discussed in relation to those of other relevant mutant types and to some actual problems of induced mutagenesis. (orig.) 891 AJ [de
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Nita, Loredana E; Chiriac, Aurica P; Bercea, Maria
2014-07-01
The present investigation was made in order to evaluate the capability of self-assembling of the two water soluble polymers, respectively, poly(aspartic acid) and Pluronic F127 into well interpenetrated mixture, and to evidence the connection effects intervened during polymer complex formation to exhibit good stability once formed, as well to understand and correlate the binding strength and the interval between better association domains. The effect of pH and temperature on the interpolymeric complex formation between poly(aspartic acid) and Pluronic F127 was studied by combining rheology with light scattering technique. The solution mixtures between poly(aspartic acid) and Pluronic F127 are Newtonian fluids for all ratios among them. Depending on the polymeric mixture composition and experimental temperature, positive or negative deviations of the experimental values from the additive dependence appear. An interesting behavior was registered around 1/1 wt. ratio between the two polymers, when the hydrodynamic diameter of the interpenetrated polymeric particles decreased suddenly. This allows us to conclude the formation of core-shell micelle structure with poly(aspartic acid) core and Pluronic F127 as shell, performed through strong interactions between polymers. This behavior was sustained by the increase of absolute value of zeta potential owing to the decrease of functional groups number at the surface of micelles. Copyright © 2014 Elsevier B.V. All rights reserved.
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Evaluation of TSPO PET Ligands [18F]VUIIS1009A and [18F]VUIIS1009B: Tracers for Cancer Imaging.
Tang, Dewei; Li, Jun; Buck, Jason R; Tantawy, Mohamed Noor; Xia, Yan; Harp, Joel M; Nickels, Michael L; Meiler, Jens; Manning, H Charles
2017-08-01
Positron emission tomography (PET) ligands targeting translocator protein (TSPO) are potential imaging diagnostics of cancer. In this study, we report two novel, high-affinity TSPO PET ligands that are 5,7 regioisomers, [ 18 F]VUIIS1009A ([ 18 F]3A) and [ 18 F]VUIIS1009B ([ 18 F]3B), and their initial in vitro and in vivo evaluation in healthy mice and glioma-bearing rats. VUIIS1009A/B was synthesized and confirmed by X-ray crystallography. Interactions between TSPO binding pocket and novel ligands were evaluated and compared with contemporary TSPO ligands using 2D 1 H- 15 N heteronuclear single quantum coherence (HSQC) spectroscopy. In vivo biodistribution of [ 18 F]VUIIS1009A and [ 18 F]VUIIS1009B was carried out in healthy mice with and without radioligand displacement. Dynamic PET imaging data were acquired simultaneously with [ 18 F]VUIIS1009A/B injections in glioma-bearing rats, with binding reversibility and specificity evaluated by radioligand displacement. In vivo radiometabolite analysis was performed using radio-TLC, and quantitative analysis of PET data was performed using metabolite-corrected arterial input functions. Imaging was validated with histology and immunohistochemistry. Both VUIIS1009A (3A) and VUIIS1009B (3B) were found to exhibit exceptional binding affinity to TSPO, with observed IC 50 values against PK11195 approximately 500-fold lower than DPA-714. However, HSQC NMR suggested that VUIIS1009A and VUIIS1009B share a common binding pocket within mammalian TSPO (mTSPO) as DPA-714 and to a lesser extent, PK11195. [ 18 F]VUIIS1009A ([ 18 F]3A) and [ 18 F]VUIIS1009B ([ 18 F]3B) exhibited similar biodistribution in healthy mice. In rats bearing C6 gliomas, both [ 18 F]VUIIS1009A and [ 18 F]VUIIS1009B exhibited greater binding potential (k 3 /k 4 )in tumor tissue compared to [ 18 F]DPA-714. Interestingly, [ 18 F]VUIIS1009B exhibited significantly greater tumor uptake (V T ) than [ 18 F]VUIIS1009A, which was attributed primarily to greater plasma
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H18F: production and use in aromatic fluorinations via triazenes
International Nuclear Information System (INIS)
Kilbourn, M.R.; Saji, H.; Welch, M.J.
1982-01-01
Studies with the triazene method of radiofluorination are presented, including the production and use of anhydrous H 18 F, investigations into the best reaction conditions, and studies of the stability and purification of the 18 F-labeled products. Despite problems with low yields, the use of triazenes in the prepartion of fluorine-18 labeled receptor ligands remains a sound synthetic approach, and the only one available for no-carrier-added syntheses. However, it appears that the fluorine-18 fluorination yields are much higher with simpler triazenes. For this reason, synthetic efforts are now focused on the preparation of 18 F-spiroperidol by a convergent synthesis
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Calculation of hyperfine constants for the H center in LiF
International Nuclear Information System (INIS)
Alencar, P.T.S.
1975-01-01
The EPR and the ENDOR hyperfine parameters for the H center in LiF are calculated assuming that the F - 2 central molecule is a simplified model for the electronic structure of this deffect. The best theoretical fitting was obtained by relaxing the ions neighboring the central molecule. We have obtained relaxations which are in agreement with the physical nature of this deffect, but for some ions the results have shown that a better wave functions for the unpaired electrons must be used. The results and the limitations of the suggested model, are discussed proposing a more realistic description for the deffect, than the F - 2 central molecule model
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International Nuclear Information System (INIS)
Xia Yingwu; Liu Guifu; Shu Qingyao; Jiang Ronghua; Xie Jiahua
1995-01-01
Chlorophyll mutants of different type were obtained from indica type temperature sensitive genie male-sterile rice (cv. 2177s) by using 60 Co γ-rays irradiation. The total chlorophyll mutation frequency reached to 0.26% in M 2 generation. However only about 4.50% of these mutants could survived. Among them, 33 heritable chlorophyll mutant lines were easily distinguished, and were screened and studied. The mutants either showed chlorosis or yellowing or expressed only at seedling period or persisted all growth cycle. The expression of mutant character was stable under different environment. It is suggested that they are useful as the marker traits in two-line hybrid rice. Moreover, the agronomic traits of most of these lines changed in different levels compared with the parent line 2177S. Every mutation line seemed to be controlled by one recessive gene as the F 1 plants of reciprocal crosses between mutant and 2177S showed normal leaf color. And the ratio of green plants/mutant plants was 3:1 in the segregated F 2 population
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Katre, Gouri; Ajmera, Namasvi; Zinjarde, Smita; RaviKumar, Ameeta
2017-10-24
Oleaginous yeasts are fast emerging as a possible feedstock for biodiesel production. Yarrowia lipolytica, a model oleaginous yeast is known to utilize a variety of hydrophobic substrates for lipid accumulation including waste cooking oil (WCO). Approaches to increase lipid content in this yeast include metabolic engineering which requires manipulation of multiple genes in the lipid biosynthesis pathway. A classical and cost-effective approach, namely, random chemical mutagenesis on the yeast can lead to increased production of biodiesel as is explored here. In this study, chemical mutagenesis using the alkylating agent, N- methyl-N'-nitro-N-nitrosoguanidine (MNNG) as well as an additional treatment with cerulenin, a fatty acid synthase inhibitor generated 800 mutants of Y. lipolytica NCIM 3589 (761 MNNG treated and 39 MNNG + cerulenin treated). A three-stage screening using Sudan Black B plate technique, Nile red fluorimetry and total lipid extraction using solvent was performed, which enabled selection of ten high lipid yielding mutants. Time course studies of all the ten mutants were further undertaken in terms of biomass, lipid yield and lipid content to select three stable mutants (YlB6, YlC7 and YlE1) capable of growing and accumulating lipid on WCO, with lipid contents of 55, 60 and 67% as compared to 45% for the wild type. The mutants demonstrated increased volumetric lipid productivities (0.062, 0.044 and 0.041 g L -1 h -1 ) as compared to the wild type (0.033 g L -1 h -1 ). The fatty acid profile of the three mutants consisted of a high content of C16 and C18 saturated and monounsaturated fatty acids and was found to be suitable for biodiesel production. The fuel properties, namely, density, kinematic viscosity, total acid number, iodine value of the three mutants were evaluated and found to lie within the limits specified by internationally accepted standards. Additionally, it was noted that the mutants demonstrated better cetane numbers and
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Directory of Open Access Journals (Sweden)
Edgard Jauregui
2017-11-01
Full Text Available The calcium/calmodulin-dependent protein kinase (CCaMK is regulated by free Ca2+ and Ca2+-loaded calmodulin. This dual binding is believed to be involved in its regulation and associated physiological functions, although direct experimental evidence for this is lacking. Here we document that site-directed mutations in the calmodulin-binding domain of CCaMK alters its binding capacity to calmodulin, providing an effective approach to study how calmodulin regulates CCaMK in terms of kinase activity and regulation of rhizobial symbiosis in Medicago truncatula. We observed that mutating the tryptophan at position 342 to phenylalanine (W342F markedly increased the calmodulin-binding capability of the mutant. The mutant CCaMK underwent autophosphorylation and catalyzed substrate phosphorylation in the absence of calcium and calmodulin. When the mutant W342F was expressed in ccamk-1 roots, the transgenic roots exhibited an altered nodulation phenotype. These results indicate that altering the calmodulin-binding domain of CCaMK could generate a constitutively activated kinase with a negative role in the physiological function of CCaMK.
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Mutant fatty acid desaturase and methods for directed mutagenesis
Shanklin, John [Shoreham, NY; Whittle, Edward J [Greenport, NY
2008-01-29
The present invention relates to methods for producing fatty acid desaturase mutants having a substantially increased activity towards substrates with fewer than 18 carbon atom chains relative to an unmutagenized precursor desaturase having an 18 carbon chain length specificity, the sequences encoding the desaturases and to the desaturases that are produced by the methods. The present invention further relates to a method for altering a function of a protein, including a fatty acid desaturase, through directed mutagenesis involving identifying candidate amino acid residues, producing a library of mutants of the protein by simultaneously randomizing all amino acid candidates, and selecting for mutants which exhibit the desired alteration of function. Candidate amino acids are identified by a combination of methods. Enzymatic, binding, structural and other functions of proteins can be altered by the method.
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Herrera, Alfa; Vu, Bao G; Stach, Christopher S; Merriman, Joseph A; Horswill, Alexander R; Salgado-Pabón, Wilmara; Schlievert, Patrick M
2016-05-03
β-Toxin is an important virulence factor of Staphylococcus aureus, contributing to colonization and development of disease [Salgado-Pabon, W., et al. (2014) J. Infect. Dis. 210, 784-792; Huseby, M. J., et al. (2010) Proc. Natl. Acad. Sci. U.S.A. 107, 14407-14412; Katayama, Y., et al. (2013) J. Bacteriol. 195, 1194-1203]. This cytotoxin has two distinct mechanisms of action: sphingomyelinase activity and DNA biofilm ligase activity. However, the distinct mechanism that is most important for its role in infective endocarditis is unknown. We characterized the active site of β-toxin DNA biofilm ligase activity by examining deficiencies in site-directed mutants through in vitro DNA precipitation and biofilm formation assays. Possible conformational changes in mutant structure compared to that of wild-type toxin were assessed preliminarily by trypsin digestion analysis, retention of sphingomyelinase activity, and predicted structures based on the native toxin structure. We addressed the contribution of each mechanism of action to producing infective endocarditis and sepsis in vivo in a rabbit model. The H289N β-toxin mutant, lacking sphingomyelinase activity, exhibited lower sepsis lethality and infective endocarditis vegetation formation compared to those of the wild-type toxin. β-Toxin mutants with disrupted biofilm ligase activity did not exhibit decreased sepsis lethality but were deficient in infective endocarditis vegetation formation compared to the wild-type protein. Our study begins to characterize the DNA biofilm ligase active site of β-toxin and suggests β-toxin functions importantly in infective endocarditis through both of its mechanisms of action.
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21 CFR 184.1635 - Potassium iodate.
2010-04-01
... ingredient is used as a dough strengthener as defined in § 170.3(o)(6) of this chapter. (d) The ingredient is used in the manufacture of bread in accordance with § 184.1(b)(2) of this chapter in an amount not to...
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Improved hydrogen storage properties of LiAlH{sub 4} by mechanical milling with TiF{sub 3}
Energy Technology Data Exchange (ETDEWEB)
Zang, Lei; Cai, Jiaxing; Zhao, Lipeng; Gao, Wenhong; Liu, Jian, E-mail: liujian@nankai.eud.cn; Wang, Yijing
2015-10-25
Dehydrogenation behavior of LiAlH{sub 4} (lithium alanate) admixed with TiF{sub 3} is investigated by pressure-composition-temperature (PCT), fourier transform infrared spectroscopy (FTIR), X-ray Diffraction (XRD), differential scanning calorimetry (DSC) and temperature programmed desorption (TPD). The TiF{sub 3} addition enhances kinetics of LiAlH{sub 4} and decreases the decomposition temperature. The LiAlH{sub 4}-2 mol % TiF{sub 3} sample starts to release hydrogen at about 35 °C and the dehydrogenation rate reaches a maximum value at 108.4 °C, compared with 145 °C and 179.9 °C for the as-received LiAlH{sub 4}. As for the dehydrogenation kinetics, the LiAlH{sub 4}-2 mol % TiF{sub 3} sample releases about 7.0 wt % H{sub 2} at 140 °C within 80 min. In comparison, the as-received LiAlH{sub 4} sample releases only 0.8 wt % hydrogen under the same conditions. The existence of proposed catalyst, Al{sub 3}Ti formed in-situ in the process of dehydrogenation, has been confirmed experimentally by XRD measurements. The activation energy of LiAlH{sub 4}-2 mol % TiF{sub 3} composite is deduced to be 66.76 kJ mol{sup −1} and 88.21 kJ mol{sup −1} for the first and second reaction stages of LiAlH{sub 4} dehydrogenation. - Highlights: • TiF{sub 3} considerably enhances the dehydrogenation kinetics of LiAlH{sub 4}. • TiF{sub 3}-doped LiAlH{sub 4} dehydrogenates even at room temperature. • Low activation energy of the dehydrogenation reaction. • Al{sub 3}Ti formed in-situ helps to explain the dehydrogenation mechanism.
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Zhao, Feng; Meng, Songsong; Zhou, Deqing
2016-02-04
To construct heptyl glycosyltransferase gene II (waaF) gene deletion mutant of Vibrio parahaemolyticus, and explore the function of the waaF gene in Vibrio parahaemolyticus. The waaF gene deletion mutant was constructed by chitin-based transformation technology using clinical isolates, and then the growth rate, morphology and serotypes were identified. The different sources (O3, O5 and O10) waaF gene complementations were constructed through E. coli S17λpir strains conjugative transferring with Vibrio parahaemolyticus, and the function of the waaF gene was further verified by serotypes. The waaF gene deletion mutant strain was successfully constructed and it grew normally. The growth rate and morphology of mutant were similar with the wild type strains (WT), but the mutant could not occurred agglutination reaction with O antisera. The O3 and O5 sources waaF gene complementations occurred agglutination reaction with O antisera, but the O10 sources waaF gene complementations was not. The waaF gene was related with O-antigen synthesis and it was the key gene of O-antigen synthesis pathway in Vibrio parahaemolyticus. The function of different sources waaF gene were not the same.
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Identification and Characterization of Spontaneous Auxotrophic Mutants in Fusarium langsethiae
Directory of Open Access Journals (Sweden)
Olga Gavrilova
2017-03-01
Full Text Available Analysis of 49 strains of Fusarium langsethiae originating from northern Europe (Russia, Finland, Sweden, UK, Norway, and Latvia revealed the presence of spontaneous auxotrophic mutants that reflect natural intraspecific diversity. Our investigations detected that 49.0% of F. langsethiae strains were auxotrophic mutants for biotin, and 8.2% of the strains required thiamine as a growth factor. They failed to grow on vitamin-free media. For both prototrophic and auxotrophic strains, no growth defect was observed in rich organic media. Without essential vitamins, a significant reduction in the growth of the auxotrophic strains results in a decrease of the formation of T-2 toxin and diacetoxyscirpenol. In addition, all analysed F. langsethiae strains were distinguished into two subgroups based on PCR product sizes. According to our results, 26 and 23 strains of F. langsethiae belong to subgroups I and II respectively. We determined that the deletion in the intergenic spacer (IGS region of the rDNA of F. langsethiae belonging to subgroup II is linked with temperature sensitivity and causes a decrease in strain growth at 30 °C. Four thiamine auxotrophic strains were found in subgroup I, while 21 biotin auxotrophic strains were detected in subgroups II. To the best of our knowledge, the spontaneous mutations in F. langsethiae observed in the present work have not been previously reported.
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Kulikova, Vitalia V; Anufrieva, Natalya V; Revtovich, Svetlana V; Chernov, Alexander S; Telegin, Georgii B; Morozova, Elena A; Demidkina, Tatyana V
2016-10-01
Pyridoxal 5'-phosphate-dependent methionine γ-lyase (MGL) catalyzes the β-elimination reaction of S-alk(en)yl-l-cysteine sulfoxides to thiosulfinates, which possess antimicrobial activity. Partial inactivation of the enzyme in the course of the reaction occurs due to oxidation of active site cysteine 115 conserved in bacterial MGLs. In this work, the C115H mutant form of Clostridium sporogenes MGL was prepared and the steady-state kinetic parameters of the enzyme were determined. The substitution results in an increase in the catalytic efficiency of the mutant form towards S-substituted l-cysteine sulfoxides compared to the wild type enzyme. We used a sulfoxide/enzyme system to generate antibacterial activity in situ. Two-component systems composed of the mutant enzyme and three S-substituted l-cysteine sulfoxides were demonstrated to be effective against Gram-positive and Gram-negative bacteria and three clinical isolates from mice. © 2016 IUBMB Life, 68(10):830-835, 2016. © 2016 International Union of Biochemistry and Molecular Biology.
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Energy Technology Data Exchange (ETDEWEB)
Jantz, Stephan G.; Hoeppe, Henning A. [Lehrstuhl fuer Festkoerperchemie, Institut fuer Physik, Universitaet Augsburg (Germany); Wuellen, Leo van; Fischer, Andreas [Lehrstuhl fuer Chemische Physik und Materialwissenschaften, Institut fuer Physik, Universitaet Augsburg (Germany); Libowitzky, Eugen [Institute for Mineralogy and Crystallography, Faculty of Geosciences, Geography and Astronomy, University of Vienna (Austria); Baran, Enrique J. [Centro de Quimica Inorganica (CEQUINOR/CONICET, UNLP), Facultad de Ciencias Exactas, Universidad Nacional de La Plata (Argentina); Weil, Matthias [Institute for Chemical Technologies and Analytics, Division Structural Chemistry, Vienna University of Technology (Austria)
2016-03-15
Single crystals of Sr(PO{sub 3}F).H{sub 2}O {P2_1/c, Z = 4, a = 7.4844(2) Aa, b = 7.0793(2) Aa, c = 8.4265(2) Aa, β = 108.696(1) , V = 422.91(2) Aa"3, 2391 F_o"2, 70 parameters, R_1[F"2 > 2σ(F"2)] = 0.036; wR_2(F"2 all) = 0.049, S = 1.054} were grown from an aqueous solution by a metathesis reaction. The structure comprises [SrO{sub 8}] polyhedra and PO{sub 3}F tetrahedra that form a layered arrangement parallel to (100). The topotactic dehydration of this phase proceeds between 80 and 140 C to afford Sr(PO{sub 3}F). The monazite-type crystal structure of Sr(PO{sub 3}F) was elucidated from the X-ray powder data by simulated annealing [P2{sub 1}/c, Z = 4, a = 6.71689(9) Aa, b = 7.11774(11) Aa, c = 8.66997(13) Aa, β = 128.0063(7) , V = 326.605(8) Aa{sup 3}, R{sub p} = 0.010, R{sub wp} = 0.015, R{sub F} = 0.030]. During dehydration, the structure of Sr(PO{sub 3}F) .H{sub 2}O collapses along [100] from a layered arrangement into a framework structure, accompanied by a change of the coordination number of the Sr{sup 2+} ions from eight to nine. The magic-angle spinning (MAS) NMR and vibrational spectroscopy data of both phases are discussed. (Copyright copyright 2016 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)
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The acid test of fluoride: how pH modulates toxicity.
Sharma, Ramaswamy; Tsuchiya, Masahiro; Skobe, Ziedonis; Tannous, Bakhos A; Bartlett, John D
2010-05-28
It is not known why the ameloblasts responsible for dental enamel formation are uniquely sensitive to fluoride (F(-)). Herein, we present a novel theory with supporting data to show that the low pH environment of maturating stage ameloblasts enhances their sensitivity to a given dose of F(-). Enamel formation is initiated in a neutral pH environment (secretory stage); however, the pH can fall to below 6.0 as most of the mineral precipitates (maturation stage). Low pH can facilitate entry of F(-) into cells. Here, we asked if F(-) was more toxic at low pH, as measured by increased cell stress and decreased cell function. Treatment of ameloblast-derived LS8 cells with F(-) at low pH reduced the threshold dose of F(-) required to phosphorylate stress-related proteins, PERK, eIF2alpha, JNK and c-jun. To assess protein secretion, LS8 cells were stably transduced with a secreted reporter, Gaussia luciferase, and secretion was quantified as a function of F(-) dose and pH. Luciferase secretion significantly decreased within 2 hr of F(-) treatment at low pH versus neutral pH, indicating increased functional toxicity. Rats given 100 ppm F(-) in their drinking water exhibited increased stress-mediated phosphorylation of eIF2alpha in maturation stage ameloblasts (pHdental fluorosis.
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Guidelines for acceptable soil concentrations in the old F- and H-Area Retention Basins
International Nuclear Information System (INIS)
Hamby, D.M.
1994-01-01
Concentration guidelines for residual radionuclides in soil at the sites of the Old F- and H-Area Retention Basins (281-3F, 281-3H) have been calculated using a dose-based approach. The guidelines also are being applied to areas around the F-Basin's Process Line. Estimation of these soil guidelines was completed using RESRAD 5.0 in accordance with the DOE RESRAD methodology specified in DOE/CH/8901 (Gi89). Guidelines are provided for the nuclides known to be present in the soils at each basin (Sc87). Soil and hydrologic characteristics specific to each basin are defined for the areas above, within, and beneath the contaminated zones
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Rahantaniaina, Marie-Sylviane; Li, Shengchun; Chatel-Innocenti, Gilles; Tuzet, Andrée; Mhamdi, Amna; Vanacker, Hélène; Noctor, Graham
2017-08-03
Glutathione is a pivotal molecule in oxidative stress, during which it is potentially oxidized by several pathways linked to H 2 O 2 detoxification. We have investigated the response and functional importance of 3 potential routes for glutathione oxidation pathways mediated by glutathione S-transferases (GST), glutaredoxin-dependent peroxiredoxins (PRXII), and dehydroascorbate reductases (DHAR) in Arabidopsis during oxidative stress. Loss-of-function gstU8, gstU24, gstF8, prxIIE and prxIIF mutants as well as double gstU8 gstU24, gstU8 gstF8, gstU24 gstF8, prxIIE prxIIF mutants were obtained. No mutant lines showed marked changes in their phenotype and glutathione profiles in comparison to the wild-type plants in either optimal conditions or oxidative stress triggered by catalase inhibition. By contrast, multiple loss of DHAR functions markedly decreased glutathione oxidation triggered by catalase deficiency. To assess whether this effect was mediated directly by loss of DHAR enzyme activity, or more indirectly by upregulation of other enzymes involved in glutathione and ascorbate recycling, we measured expression of glutathione reductase (GR) and expression and activity of monodehydroascorbate reductases (MDHAR). No evidence was obtained that either GRs or MDHARs were upregulated in plants lacking DHAR function. Hence, interplay between different DHARs appears to be necessary to couple ascorbate and glutathione pools and to allow glutathione-related signaling during enhanced H 2 O 2 metabolism.
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A sorghum (Sorghum bicolor mutant with altered carbon isotope ratio.
Directory of Open Access Journals (Sweden)
Govinda Rizal
Full Text Available Recent efforts to engineer C4 photosynthetic traits into C3 plants such as rice demand an understanding of the genetic elements that enable C4 plants to outperform C3 plants. As a part of the C4 Rice Consortium's efforts to identify genes needed to support C4 photosynthesis, EMS mutagenized sorghum populations were generated and screened to identify genes that cause a loss of C4 function. Stable carbon isotope ratio (δ13C of leaf dry matter has been used to distinguishspecies with C3 and C4 photosynthetic pathways. Here, we report the identification of a sorghum (Sorghum bicolor mutant with a low δ13C characteristic. A mutant (named Mut33 with a pale phenotype and stunted growth was identified from an EMS treated sorghum M2 population. The stable carbon isotope analysis of the mutants showed a decrease of 13C uptake capacity. The noise of random mutation was reduced by crossing the mutant and its wildtype (WT. The back-cross (BC1F1 progenies were like the WT parent in terms of 13C values and plant phenotypes. All the BC1F2 plants with low δ13C died before they produced their 6th leaf. Gas exchange measurements of the low δ13C sorghum mutants showed a higher CO2 compensation point (25.24 μmol CO2.mol-1air and the maximum rate of photosynthesis was less than 5μmol.m-2.s-1. To identify the genetic determinant of this trait, four DNA pools were isolated; two each from normal and low δ13C BC1F2 mutant plants. These were sequenced using an Illumina platform. Comparison of allele frequency of the single nucleotide polymorphisms (SNPs between the pools with contrasting phenotype showed that a locus in Chromosome 10 between 57,941,104 and 59,985,708 bps had an allele frequency of 1. There were 211 mutations and 37 genes in the locus, out of which mutations in 9 genes showed non-synonymous changes. This finding is expected to contribute to future research on the identification of the causal factor differentiating C4 from C3 species that can be used
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Gouider, Mbarka; Feki, Mongi; Sayadi, Sami
2009-10-30
Fluoride content and flow-rate of fertilizer plant wastewater from phosphoric acid and/or triple superphosphate (TSP) production lead to the discharge of several thousand tons of fluoride (F(-)) per year and even more for phosphate (PO4(3-)). Since sustainability is an important environmental concern, the removal methods should allow phosphorus and fluoride to be recycled as a sustainable products for use as raw materials either in agricultural or industrial applications. In the present work, separative recovery with lime of these two target species was investigated. A preliminary speciation study, carried out on the crude effluent, showed that two forms of fluoride: HF and H2SiF6 are present in a highly acidic medium (pH approximately 2). Evidence that fluoride is present under both free (HF) and combined (H2SiF6) forms, in the phosphate-containing effluent, was provided by comparing potentiometric titration curves of a crude wastewater sample and synthetic acid mixtures containing H3PO4, HF and H2SiF6. In a second step synthetic effluent containing mixtures of the following acids: HF, H2SiF6 and H3PO4, were treated with lime. The behaviour of these compounds under lime treatment was analysed. The data showed that fluoride has a beneficial effect on phosphate removal. Moreover, by acting on the precipitation pH, a "selective" recovery of fluoride and phosphate ions was possible either from phosphoric acid/hydrofluoric acid or phosphoric acid/hexafluorosilicic acid mixtures. Indeed, the first stage of the separative recovery, led to a fluoride removal efficiency of 97-98% from phosphoric acid/hydrofluoric acid mixture. It was of 93-95% from phosphoric acid/hexafluorosilicic acid mixture. During the second stage, the phosphate precipitation reached 99.8% from both acidic mixtures whereas it did not exceed 82% from a solution containing H3PO4 alone. The XRD and IR analyses showed that during lime treatment, a H2SiF6 hydrolysis occurred, instead of CaSiF6 solid
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Theoretical Analysis of Allosteric and Operator Binding for Cyclic-AMP Receptor Protein Mutants
Einav, Tal; Duque, Julia; Phillips, Rob
2018-02-01
Allosteric transcription factors undergo binding events both at their inducer binding sites as well as at distinct DNA binding domains, and it is often difficult to disentangle the structural and functional consequences of these two classes of interactions. In this work, we compare the ability of two statistical mechanical models - the Monod-Wyman-Changeux (MWC) and the Koshland-N\\'emethy-Filmer (KNF) models of protein conformational change - to characterize the multi-step activation mechanism of the broadly acting cyclic-AMP receptor protein (CRP). We first consider the allosteric transition resulting from cyclic-AMP binding to CRP, then analyze how CRP binds to its operator, and finally investigate the ability of CRP to activate gene expression. In light of these models, we examine data from a beautiful recent experiment that created a single-chain version of the CRP homodimer, thereby enabling each subunit to be mutated separately. Using this construct, six mutants were created using all possible combinations of the wild type subunit, a D53H mutant subunit, and an S62F mutant subunit. We demonstrate that both the MWC and KNF models can explain the behavior of all six mutants using a small, self-consistent set of parameters. In comparing the results, we find that the MWC model slightly outperforms the KNF model in the quality of its fits, but more importantly the parameters inferred by the MWC model are more in line with structural knowledge of CRP. In addition, we discuss how the conceptual framework developed here for CRP enables us to not merely analyze data retrospectively, but has the predictive power to determine how combinations of mutations will interact, how double mutants will behave, and how each construct would regulate gene expression.
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Elizalde, María Mercedes; Sevic, Ina; González López Ledesma, María Mora; Campos, Rodolfo Héctor; Barbini, Luciana; Flichman, Diego Martin
2018-01-01
In the context of pathogenesis of HBV infection, HBV genotypes and mutants have been shown to affect the natural course of chronic infection and treatment outcomes. In this work, we studied the induction of apoptosis by the replication of HBV subgenotypes F1b and F4, and the naturally occurring mutants BCP and preCore. Both subgenotypes F1b and F4 HBV genome transfections induced cell death by apoptosis in human hepatocytes. The BCPdm (A1762T/G1764A) and preCore (G1896A) mutants induced higher levels of apoptosis than the wt virus. This increase in apoptosis was not associated with the enhanced viral replication of the variants. HBV-mediated apoptosis was independent of viral subgenotypes, and associated with the modulation of members of the regulatory Bcl-2 family proteins expression in the mitochondrial apoptotic pathway. Finally, the apoptosis induction increase observed for the preCore mutants suggests that HBeAg might have an anti-apoptotic effect in human hepatocytes. Copyright © 2017 Elsevier Inc. All rights reserved.
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Chang, Chih-Kang; Teng, Kuo-Hsun; Lin, Sheng-Wei; Chang, Tao-Hsin; Liang, Po-Huang
2013-04-23
Previously we showed that yeast geranylgeranyl diphosphate synthase (GGPPS) becomes an inactive monomer when the first N-terminal helix involved in dimerization is deleted. This raises questions regarding why dimerization is required for GGPPS activity and which amino acids in the dimer interface are essential for dimerization-mediated activity. According to the GGPPS crystal structure, three amino acids (N101, N104, and Y105) located in the helix F of one subunit are near the active site of the other subunit. As presented here, when these residues were replaced individually with Ala caused insignificant activity changes, N101A/Y105A and N101A/N104A but not N104A/Y105A showed remarkably decreased k(cat) values (200-250-fold). The triple mutant N101A/N104A/Y105A displayed no detectable activity, although dimer was retained in these mutants. Because N101 and Y105 form H-bonds with H139 and R140 in the other subunit, respectively, we generated H139A/R140A double mutant and found it was inactive and became monomeric. Therefore, the multiple mutations apparently influence the integrity of the catalytic site due to the missing H-bonding network. Moreover, Met111, also on the highly conserved helix F, was necessary for dimer formation and enzyme activity. When Met111 was replaced with Glu, the negative-charged repulsion converted half of the dimer into a monomer. In conclusion, the H-bonds mainly through N101 for maintaining substrate binding stability and the hydrophobic interaction of M111 in dimer interface are essential for activity of yeast GGPPS.
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Deventhiran, Jagadeeswaran; Kumar, Sandeep R P; Raghunath, Shobana; Leroith, Tanya; Elankumaran, Subbiah
2016-01-01
PB1-F2 protein, the 11th influenza A virus (IAV) protein, is considered to play an important role in primary influenza virus infection and postinfluenza secondary bacterial pneumonia in mice. The functional role of PB1-F2 has been reported to be a strain-specific and host-specific phenomenon. Its precise contribution to the pathogenicity and transmission of influenza virus in mammalian host, such as swine, and avian hosts, such as turkeys, remain largely unknown. In this study, we explored the role of PB1-F2 protein of triple-reassortant (TR) H3N2 swine influenza virus (SIV) in pigs and turkeys. Using the eight-plasmid reverse genetics system, we rescued wild-type SIV A/swine/Minnesota/1145/2007 (H3N2) (SIV 1145-WT), a PB1-F2 knockout mutant (SIV 1145-KO), and its N66S variant (SIV 1145-N66S). The ablation of PB1-F2 in SIV 1145 modulated early-stage apoptosis but did not affect the viral replication in swine alveolar macrophage cells. In pigs, PB1-F2 expression did not affect nasal shedding, lung viral load, immunophenotypes, and lung pathology. On the other hand, in turkeys, SIV 1145-KO infected poults, and its in-contacts developed clinical signs earlier than SIV 1145-WT groups and also displayed more extensive histopathological changes in intestine. Further, turkeys infected with SIV 1145-N66S displayed poor infectivity and transmissibility. The more extensive histopathologic changes in intestine and relative transmission advantage observed in turkeys infected with SIV 1145-KO need to be further explored. Taken together, these results emphasize the host-specific roles of PB1-F2 in the pathogenicity and transmission of IAV. Novel triple-reassortant H3N2 swine influenza virus emerged in 1998 and spread rapidly among the North American swine population. Subsequently, it showed an increased propensity to reassort, generating a range of reassortants. Unlike classical swine influenza virus, TR SIV produces a full-length PB1-F2 protein, which is considered an important
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Directory of Open Access Journals (Sweden)
Hassan Abdel-Latif A. Mohamed
2006-06-01
Full Text Available Exposing a wild-type culture of Trichoderma harzianum to gamma irradiation induced two stable salt-tolerant mutants (Th50M6 and Th50M11. Under saline conditions, both mutants greatly surpassed their wild type strain in growth rate, sporulation and biological proficiency against Fusarium oxysporum, the causal agent of tomato wilt disease. Tolerant T. harzianum mutants detained a capability to grow and convinced sporulation in growth media containing up to 69 mM NaCl. In comparison with their parent strain, characterization of both mutants confirmed that they have reinforced contents of proline and hydroxyproline, relatively higher sodium content compared to potassium, calcium or magnesium contents, higher level of total phenols. Electrophoretic analysis of total soluble proteins in the salt tolerance mutant Th50M6 showed different bands accumulated in response to 69 mM NaCl. Data also showed that mutants produce certain active metabolites, such as chitinases, cellulases, beta-galactosidases, as well as, some antibiotics i.e., trichodermin, gliotoxin and gliovirin. Trichoderma mutants significantly reduced wilt disease incidence and improved yield and mineral contents of tomato plants under both saline and non-saline soil conditions, as well as, under infested and natural conditions. T. harzianum mutants were also more efficient in dropping the F. oxysporum growth in rhizosphere compared to the wild type strain. Population density of both mutants in rhizosphere far exceeded that of T. harzianum wild type strain.A exposição de uma cepa selvagem de Trichoderma harzianum à irradiação gama induziu dois mutantes tolerantes a sal (Th50M6 e Th50M11. Em condições salinas, os dois mutantes foram muito superiores à cepa selvagem em relação à velocidade de multiplicação, esporulação e eficiência contra Fusarium oxysporum, o agente causador da doença wilt do tomate. Os mutantes tolerantes foram capazes de multiplicação e esporulação em
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International Nuclear Information System (INIS)
Capron, J.M.
2008-01-01
The 100-F-26:9 underground pipeline subsite consists of the sanitary sewers servicing the 105-F, 108-F, 184-F, 185-F, and 190-F buildings, and the 1700-F administration and service buildings (1704-F, 1707-F, 1707-FA, 1713-F, 1717-F, 1719-F, and 1722-F). In accordance with this evaluation, the confirmatory and verification sampling results support a reclassification of this site to Interim Closed Out. The current site conditions achieve the remedial action objectives and the corresponding remedial action goals established in the Remaining Sites ROD. The results of verification sampling show that residual contaminant concentrations do not preclude any future uses and allow for unrestricted use of shallow zone soils. The results also demonstrate that residual contaminant concentrations are protective of groundwater and the Columbia River
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Short communication. A spontaneous mutant of L-202 rice
Energy Technology Data Exchange (ETDEWEB)
Garcia-Yzaguire, A.; Padrones, T.
2009-07-01
A new spontaneous phenotype of the rice cultivar L-202 was found. Mendelian analysis indicates that it is a monogenic, recessive mutant. Its distinguishing features are: dark blue-green colour, short and narrow leaves, high tillering and relatively short height. The objectives of this study were: to characterize it, to determine if it is heritable and if so, its genetic basis. Its distinguishing features are: dark blue-green colour, short and narrow leaves, high tillering and relatively short height. Selfing the new phenotype resulted in a uniform progeny, with the traits of the parent plant (wild type). Crossing the new phenotype with the normal L-202 cultivar resulted in a uniform F1 hybrid generation, with the wild type. The F2 generation showed a mendelian segregation which did not depart significantly from three normal plants : one new phenotype. It is concluded that it is a monogenic, recessive mutant. (Author) 3 refs.
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Friedman, Adam D; Kim, Dongwook; Liu, Rihe
2015-01-01
When developed as targeting ligands for the in vivo delivery of biomaterials to biological systems, RNA aptamers immediately face numerous obstacles, in particular nuclease degradation and post-selection 2' modification. This study aims to develop a novel class of highly stable, 2'-fully modified RNA aptamers that are ideal for the targeted delivery of biomaterials. We demonstrated the facile transcription of a fGmH (2'-F-dG, 2'-OMe-dA/dC/dU) RNA library with unexpected hydrophobicity, the direct selection of aptamers from a fGmH RNA library that bind Staphylococcus aureus Protein A (SpA) as a model target, and the superior nuclease and serum stability of these aptamers compared to 2'-partially modified RNA variants. Characterizations of fGmH RNA aptamers binding to purified SpA and to endogenous SpA present on the surface of S. aureus cells demonstrate fGmH RNA aptamer selectivity and stability. Significantly, fGmH RNA aptamers were able to functionalize, stabilize, and specifically deliver aggregation-prone silver nanoparticles (AgNPs) to S. aureus with SpA-dependent antimicrobial effects. This study describes a novel aptamer class with considerable potential to improve the in vivo applicability of nucleic acid-based affinity molecules to biomaterials.
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International Nuclear Information System (INIS)
Zhang, Yanfeng; Gao, Xiaoli
2011-01-01
Recombinant wild-type l-lactate dehydrogenase from B. subtilis (BsLDH) was cocrystallized with fructose 1,6-bisphosphate and NAD + and the crystal diffracted to 2.38 Å resolution. The H171C mutant of BsLDH was also crystallized as the apoenzyme and in complex with NAD + and the crystals diffracted to 2.20 and 2.49 Å, respectively. All crystals belonged to space group P3. l-Lactate dehydrogenase (LDH) is an important enzyme involved in the last step of glycolysis that catalyzes the reversible conversion of pyruvate to l-lactate with the simultaneous oxidation of NADH to NAD + . In this study, wild-type LDH from Bacillus subtilis (BsLDH-WT) and the H171C mutant (BsLDH-H171C) were expressed in Escherichia coli and purified to near-homogeneity. BsLDH-WT was crystallized in the presence of fructose 1,6-bisphosphate (FBP) and NAD + and the crystal diffracted to 2.38 Å resolution. The crystal belonged to space group P3, with unit-cell parameters a = b = 171.04, c = 96.27 Å. BsLDH-H171C was also crystallized as the apoenzyme and in complex with NAD + , and data sets were collected to 2.20 and 2.49 Å resolution, respectively. Both BsLDH-H171C crystals belonged to space group P3, with unit-cell parameters a = b = 133.41, c = 99.34 Å and a = b = 133.43, c = 99.09 Å, respectively. Tetramers were observed in the asymmetric units of all three crystals
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Claudia Marcelloni
2006-01-01
Tour of the stands at the UK@CERN industrial exhibition with the Director-General and H.E. Mr Simon Featherstone, HM Ambassador to Switzerland, hosted by Mrs. Jan Fillingham MBE, Head of Exhibitions, BEAMA
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Genetic analysis of relative traits for a drought-sensitive mutant
International Nuclear Information System (INIS)
Gao Kangning; Wang Huaqi
2009-01-01
A drought-sensitive mutant (M616), selected from Handao 616 (HD616) by 60 Co γ-irradiation at 200Gy, was used. Some morphological and yield related traits of M166 and HD616 related to drought resistance were investigated. We further developed F 1 and F 2 reciprocal hybrid combinations derived from the crosses between M616 and HD616, between M616 and IRAT109, respectively, and genetic analysis of 3 traits including plant height, culm width of main stem and seed setting rate on main panicle were camed out. The results showed that M616 showed obviously sensitive to drought stress. Analysis of variance for three traits in upland and paddy between F 1 reciprocal hybrid combinations showed that each trait had no significant difference, and indicated that there were no differences of cytoplasmic hereditary effect. In addition, 3 traits of F 2 populations, were found that the frequency distributions of culm width showed normal distribution, indicating that culm width was polygenic trait, and the frequency distribution of plant height and seed setting rate did not show normal distribution, indicating that the two traits were qualitative-quantitative traits. Genetic analysis of relative traits for a drought-sensitive mutant of upland rice was a basic work for the gene location and cloning. (authors)
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McClintock, Maria K; Kaznessis, Yiannis N; Hackel, Benjamin J
2016-02-01
Vancomycin-resistant Enterococci infections are a significant clinical problem. One proposed solution is to use probiotics, such as lactic acid bacteria, to produce antimicrobial peptides at the site of infection. Enterocin A, a class 2a bacteriocin, exhibits inhibitory activity against E. faecium and E. faecalis, which account for 86% of vancomycin-resistant Enterococci infections. In this study, we aimed to engineer enterocin A mutants with enhanced potency within a lactic acid bacterial production system. Peptide mutants resulting from saturation mutagenesis at sites A24 and T27 were efficiently screened in a 96-well plate assay for inhibition of pathogen growth. Several mutants exhibit increased potency relative to wild-type enterocin A in both liquid- and solid-medium growth assays. In particular, A24P and T27G exhibit enhanced inhibition of multiple strains of E. faecium and E. faecalis, including clinically isolated vancomycin-resistant strains. A24P and T27G enhance killing of E. faecium 8 by 13 ± 3- and 18 ± 4-fold, respectively. The engineered enterocin A/lactic acid bacteria systems offer significant potential to combat antibiotic-resistant infections. © 2015 Wiley Periodicals, Inc.
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Chen, Sixue; Glawischnig, Erich; Jørgensen, Kirsten; Naur, Peter; Jørgensen, Bodil; Olsen, Carl-Erik; Hansen, Carsten H; Rasmussen, Hasse; Pickett, John A; Halkier, Barbara A
2003-03-01
Cytochromes P450 of the CYP79 family catalyze the conversion of amino acids to oximes in the biosynthesis of glucosinolates, a group of natural plant products known to be involved in plant defense and as a source of flavor compounds, cancer-preventing agents and bioherbicides. We report a detailed biochemical analysis of the substrate specificity and kinetics of CYP79F1 and CYP79F2, two cytochromes P450 involved in the biosynthesis of aliphatic glucosinolates in Arabidopsis thaliana. Using recombinant CYP79F1 and CYP79F2 expressed in Escherichia coli and Saccharomyces cerevisiae, respectively, we show that CYP79F1 metabolizes mono- to hexahomomethionine, resulting in both short- and long-chain aliphatic glucosinolates. In contrast, CYP79F2 exclusively metabolizes long-chain elongated penta- and hexahomomethionines. CYP79F1 and CYP79F2 are spatially and developmentally regulated, with different gene expression patterns. CYP79F2 is highly expressed in hypocotyl and roots, whereas CYP79F1 is strongly expressed in cotyledons, rosette leaves, stems, and siliques. A transposon-tagged CYP79F1 knockout mutant completely lacks short-chain aliphatic glucosinolates, but has an increased level of long-chain aliphatic glucosinolates, especially in leaves and seeds. The level of long-chain aliphatic glucosinolates in a transposon-tagged CYP79F2 knockout mutant is substantially reduced, whereas the level of short-chain aliphatic glucosinolates is not affected. Biochemical characterization of CYP79F1 and CYP79F2, and gene expression analysis, combined with glucosinolate profiling of knockout mutants demonstrate the functional role of these enzymes. This provides valuable insights into the metabolic network leading to the biosynthesis of aliphatic glucosinolates, and into metabolic engineering of altered aliphatic glucosinolate profiles to improve nutritional value and pest resistance.
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Mutation of purD and purF genes further attenuates Brucella abortus strain RB51.
Truong, Quang Lam; Cho, Youngjae; Barate, Abhijit Kashinath; Kim, Suk; Watarai, Masahisa; Hahn, Tae-Wook
2015-02-01
In the present study, transposon mutagenesis was used to further attenuate Brucella abortus RB51 vaccine strain. Two purD and purF mutants were constructed, characterized and evaluated for attenuation via intracellular survival in murine macrophage-like RAW264.7 and HeLa cells, and by clearance in BALB/c mice. The purD and purF mutants showed significantly decreased intracellular survival, and complementation of these mutants with intact copies of purD or purF genes of RB51 strain was able to restore these defects. In addition, the pur mutants presented significantly lowered persistence in mice. Immunization with purD and purF mutants protected mice against a challenge with the virulent B. abortus strain 544 at a level similar to that of the parent RB51. These data suggest that genes encoding the early stages of purine biosynthesis (purD and purF) are required for intracellular survival and virulence of B. abortus. Copyright © 2014 Elsevier Ltd. All rights reserved.
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Tuck, S F; Graham-Lorence, S; Peterson, J A; Ortiz de Montellano, P R
1993-01-05
Ferricyanide oxidation of the aryl-iron complexes formed by the reaction of cytochrome P450 enzymes with arylhydrazines causes in situ migration of the aryl group from the iron to the porphyrin nitrogen atoms. The regiochemistry of this migration, defined by the ratio of the four possible N-arylprotoporphyrin IX isomers, provides a method for mapping the topologies of cytochrome P450 active sites. The method has been validated by using it to examine the active site of cytochrome P450cam (CYP101), for which a crystal structure is available. In agreement with the crystal structure, reaction with phenylhydrazine gives a 5:25:70 ratio of the NA:NC:ND (subscript indicates pyrrole ring) N-phenylprotoporphyrin IX isomers. Naphthylhydrazine, however, yields exclusively the NC regioisomer and 4-(phenyl)phenylhydrazine the NA:NC:ND isomers in a 14:40:46 ratio. These isomer ratio differences are readily explained by topological differences between the upper and lower reaches of the active site. Having validated the aryl-iron shift as a topological probe, we used it to investigate the structural changes caused by mutation of Phe-87, a residue that provides the ceiling over pyrrole ring D in the crystal structure of cytochrome P450cam. Mutation of Phe-87 to a tryptophan causes no detectable change in the regiochemistry of camphor hydroxylation and only minor changes in the N-aryl isomer ratios. However, mutation of Phe-87 to an alanine, which was expected to open up the region above pyrrole ring D, severely decreased the proportion of the ND in favor of the NA isomer. Less rather than more space is therefore available over pyrrole ring D in the F87A mutant despite the fact that the regiochemistry of camphor hydroxylation remains unchanged. These results provide evidence for significant structural reorganization in the upper regions of the substrate binding site without alteration of the camphor hydroxylation regiospecificity in the F87A mutant.
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Deriving a blood-mimicking fluid for particle image velocimetry in Sylgard-184 vascular models.
Yousif, Majid Y; Holdsworth, David W; Poepping, Tamie L
2009-01-01
A new blood-mimicking fluid (BMF) has been developed for particle image velocimetry (PIV), which enables flow studies in vascular models (phantoms). A major difficulty in PIV that affects measurement accuracy is the refraction and distortion of light passing through the interface between the model and the fluid, due to the difference in refractive index (n) between the two materials. The problem can be eliminated by using a fluid with a refractive index matching that of the model. Such fluids are not commonly available, especially for vascular research where the fluid should also have a viscosity similar to human blood. In this work, a blood-mimicking fluid, composed of water (47.38% by weight), glycerol (36.94% by weight) and sodium iodide salt (15.68% by weight), was developed for compatibility with our silicone (Sylgard 184; n = 1.414) phantoms. The fluid exhibits a dynamic viscosity of 4.31+/-0.03 cP which lies within the range of human blood viscosity (4.4+/-0.6 cP). Both refractive index and viscosity were attained at 22.2+/-0.2 degrees C, which is a feasible room temperature, thus eliminating the need for a temperature-control system. The fluid will be used to study hemodynamics in vascular flow models fabricated from Sylgard 184.
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Energy Technology Data Exchange (ETDEWEB)
Bergere, R; Veyssiere, A; Daujat, P [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires
1966-06-01
This paper contains a digest of a series of studies on the frequency modulation of U.H.F. fields and signals associated with the linear electron accelerator at Saclay. We first consider the frequency modulation of a U. H. F. pulse before its injection into an accelerating structure and after its subsequent propagation when no accelerated electrons are present. We then apply a similar analysis to the frequency modulation due to the direct interaction of the electron beam itself, and the accelerating U.H.F. fields. Finally we consider the phase modulation of the elementary electron packet itself. This phase modulation can be correctly interpreted by considering the dynamics of the electron beam as such. This analysis moreover, gives a correct interpretation of the evolution of the phase modulation with time, as the elementary electron packets move along with the sinusoidal U.H.F. accelerating fields. (authors) [French] Cet article resume les etudes faites sur l'accelerateur lineaire d'electrons de Saclay a propos de la modulation de frequence des divers signaux U.H.F. presents autour de l'accelerateur. On etudie d'abord la modulation de frequence des impulsions U.H.F. entrant sur la structure acceleratrice ou transmises par cette structure en l'absence de faisceau d'electrons acceleres. On analyse ensuite la modulation de frequence resultant de l'interaction d'une de ces ondes avec le faisceau d'electrons acceleres. On etudie enfin, la modulation de phase des divers paquets elementaires constituant une impulsion d'electrons acceleres. On montre comment cette modulation de phase peut s'expliquer par des considerations sur la dynamique du faisceau et conduire a une representation dans les divers cas possibles de l'evolution de la phase d'accrochage des electrons sur l'onde sinusoidale progressive de champ accelerateur. (auteurs)
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Energy Technology Data Exchange (ETDEWEB)
Hawkins, Andrew S.; Howard, Brandon A. [Div. of Nuclear Medicine, Dept. of Radiology, Duke University Medical Center, Durham (United States)
2017-09-15
Three males aged 71 to 80 years with known stage IV metastatic prostate cancer underwent F-18 sodium fluoride (NaF) PET/CT to assess osseous metastatic disease burden and stability. In addition to F-18 NaF avid known osseous metastases, each patient also exhibited increased F-18 NaF activity in soft tissues. The first patient exhibited multiple F-18 NaF avid enlarged retroperitoneal and pelvic lymph nodes on consecutive PET/CT scans. The second patient demonstrated an F-18 NaF avid thyroid nodule on consecutive PET/CT scans. The third patient exhibited increased F-18 NaF activity in a hepatic metastasis.
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Abnormal grooming activity in Dab1(scm) (scrambler) mutant mice.
Strazielle, C; Lefevre, A; Jacquelin, C; Lalonde, R
2012-07-15
Dab1(scm) mutant mice, characterized by cell ectopias and degeneration in cerebellum, hippocampus, and neocortex, were compared to non-ataxic controls for different facets of grooming caused by brief water immersions, as well as some non-grooming behaviors. Dab1(scm) mutants were strongly affected in their quantitative functional parameters, exhibiting higher starting latencies before grooming relative to non-ataxic littermates of the A/A strain, fewer grooming bouts, and grooming components of shorter duration, with an unequal regional distribution targeting almost totally the rostral part (head washing and forelimb licking) of the animal. Only bouts of a single grooming element were preserved. The cephalocaudal order of grooming elements appeared less disorganized, mutant and control mice initiating the grooming with head washing and forelimb licking prior to licking posterior parts. However, mutants differed from controls in that all their bouts were incomplete but uninterrupted, although intergroup difference for percentage of the incorrect transitions was not significant. In contrast to grooming, Dab1(scm) mice ambulated for a longer time. During walking episodes, they exhibited more body scratching than controls, possibly to compensate for the lack of licking different body parts. In conjunction with studies with other ataxic mice, these results indicate that the cerebellar cortex affects grooming activity and is consequently involved in executing various components, but not in its sequential organization, which requires other brain regions such as cerebral cortices or basal ganglia. Copyright © 2012 Elsevier B.V. All rights reserved.
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Connexin mutants and cataracts
Directory of Open Access Journals (Sweden)
Eric C Beyer
2013-04-01
Full Text Available The lens is a multicellular, but avascular tissue that must stay transparent to allow normal transmission of light and focusing of it on the retina. Damage to lens cells and/or proteins can cause cataracts, opacities that disrupt these processes. The normal survival of the lens is facilitated by an extensive network of gap junctions formed predominantly of connexin46 and connexin50. Mutations of the genes that encode these connexins (GJA3 and GJA8 have been identified and linked to inheritance of cataracts in human families and mouse lines. In vitro expression studies of several of these mutants have shown that they exhibit abnormalities that may lead to disease. Many of the mutants reduce or modify intercellular communication due to channel alterations (including loss of function or altered gating or due to impaired cellular trafficking which reduces the number of gap junction channels within the plasma membrane. However, the abnormalities detected in studies of other mutants suggest that they cause cataracts through other mechanisms including gain of hemichannel function (leading to cell injury and death and formation of cytoplasmic accumulations (that may act as light scattering particles. These observations and the anticipated results of ongoing studies should elucidate the mechanisms of cataract development due to mutations of lens connexins and abnormalities of other lens proteins. They may also contribute to our understanding of the mechanisms of disease due to connexin mutations in other tissues.
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PNRI mutant variety: Cordyline 'Afable'
International Nuclear Information System (INIS)
Aurigue, Fernando B.
2012-01-01
Cordyline 'Afable', registered by the Philippine Nuclear Research Institute as NSIC 2009 Or-83, is an induced mutant developed from Cordyline 'Kiwi' by treating stem cuttings with acute gamma radiation from a Cobalt-60 source. The new mutant is identical to Cordyline 'Kiwi' in growth habit but differs in foliage color, and exhibits field resistance to Phytophthora sp., a fungus that causes leaf blight and rot in Ti plants. Results of this mutation breeding experiment showed that leaf color was altered by gamma irradiation and resistance to fungal diseases was improved. It also demonstrated how mutations that occur in nature may be generated artificially. Propagation of cordyline 'Afable' is true-to-type by vegetative propagation methods, such as separation of suckers and offshoots, shoot tip cutting, and top cutting. Aside from landscaping material, terrarium or dish-garden plant, it is ideal as containerized plant for indoor and outdoor use. The leaves or shoots may be harvested as cut foliage for flower arrangements. (author)