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Sample records for thermus caldophilus gk24

  1. Integrable deformations of the Gk1 ×Gk2 /Gk1+k2 coset CFTs

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    Sfetsos, Konstantinos; Siampos, Konstantinos

    2018-02-01

    We study the effective action for the integrable λ-deformation of the Gk1 ×Gk2 /Gk1+k2 coset CFTs. For unequal levels theses models do not fall into the general discussion of λ-deformations of CFTs corresponding to symmetric spaces and have many attractive features. We show that the perturbation is driven by parafermion bilinears and we revisit the derivation of their algebra. We uncover a non-trivial symmetry of these models parametric space, which has not encountered before in the literature. Using field theoretical methods and the effective action we compute the exact in the deformation parameter β-function and explicitly demonstrate the existence of a fixed point in the IR corresponding to the Gk1-k2 ×Gk2 /Gk1 coset CFTs. The same result is verified using gravitational methods for G = SU (2). We examine various limiting cases previously considered in the literature and found agreement.

  2. Integrable deformations of the GkGk2/Gk1+k2 coset CFTs

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    Konstantinos Sfetsos

    2018-02-01

    Full Text Available We study the effective action for the integrable λ-deformation of the GkGk2/Gk1+k2 coset CFTs. For unequal levels theses models do not fall into the general discussion of λ-deformations of CFTs corresponding to symmetric spaces and have many attractive features. We show that the perturbation is driven by parafermion bilinears and we revisit the derivation of their algebra. We uncover a non-trivial symmetry of these models parametric space, which has not encountered before in the literature. Using field theoretical methods and the effective action we compute the exact in the deformation parameter β-function and explicitly demonstrate the existence of a fixed point in the IR corresponding to the Gk1−k2×Gk2/Gk1 coset CFTs. The same result is verified using gravitational methods for G=SU(2. We examine various limiting cases previously considered in the literature and found agreement.

  3. Thermus arciformis sp. nov., a thermophilic species from a geothermal area.

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    Zhang, Xin-Qi; Ying, Yi; Ye, Ying; Xu, Xue-Wei; Zhu, Xu-Fen; Wu, Min

    2010-04-01

    Two aerobic, Gram-negative, non-motile, non-sporulating, yellow-pigmented bacteria, strains TH92(T) and TH91, were isolated from a hot spring located in Laibin, Guangxi, in the south-eastern geothermal area of China. The isolates grew at 40-77 degrees C (optimally at 70 degrees C) and at pH 6.0-9.5 (optimally at pH 7.5-8.0). Phylogenetic analysis of 16S rRNA gene sequences and levels of DNA-DNA relatedness together indicated that the new isolates represented a novel species of the genus Thermus with closest affinity to Thermus aquaticus, Thermus igniterrae and Thermus thermophilus. Compared with their closest relatives, strains TH92( T) and TH91 were able to assimilate a wider range of carbohydrates, amino acids and organic acids as sole carbon sources for growth, such as lactose and melibiose. The new isolates had lower combined levels of C(16 : 0 ) and iso-C(16 : 0) compared with their closest relatives. On the basis of polyphasic taxonomic characterization, strains TH92(T) and TH91 are considered to represent a single novel species of the genus Thermus, for which the name Thermus arciformis sp. nov. is proposed. The type strain is TH92(T) (=CGMCC 1.6992(T) =JCM 15153(T)).

  4. Sequence of the hyperplastic genome of the naturally competent Thermus scotoductus SA-01

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    Gounder Kamini

    2011-11-01

    Full Text Available Abstract Background Many strains of Thermus have been isolated from hot environments around the world. Thermus scotoductus SA-01 was isolated from fissure water collected 3.2 km below surface in a South African gold mine. The isolate is capable of dissimilatory iron reduction, growth with oxygen and nitrate as terminal electron acceptors and the ability to reduce a variety of metal ions, including gold, chromate and uranium, was demonstrated. The genomes from two different Thermus thermophilus strains have been completed. This paper represents the completed genome from a second Thermus species - T. scotoductus. Results The genome of Thermus scotoductus SA-01 consists of a chromosome of 2,346,803 bp and a small plasmid which, together are about 11% larger than the Thermus thermophilus genomes. The T. thermophilus megaplasmid genes are part of the T. scotoductus chromosome and extensive rearrangement, deletion of nonessential genes and acquisition of gene islands have occurred, leading to a loss of synteny between the chromosomes of T. scotoductus and T. thermophilus. At least nine large inserts of which seven were identified as alien, were found, the most remarkable being a denitrification cluster and two operons relating to the metabolism of phenolics which appear to have been acquired from Meiothermus ruber. The majority of acquired genes are from closely related species of the Deinococcus-Thermus group, and many of the remaining genes are from microorganisms with a thermophilic or hyperthermophilic lifestyle. The natural competence of Thermus scotoductus was confirmed experimentally as expected as most of the proteins of the natural transformation system of Thermus thermophilus are present. Analysis of the metabolic capabilities revealed an extensive energy metabolism with many aerobic and anaerobic respiratory options. An abundance of sensor histidine kinases, response regulators and transporters for a wide variety of compounds are indicative

  5. Complete Genome Analysis of Thermus parvatiensis and Comparative Genomics of Thermus spp. Provide Insights into Genetic Variability and Evolution of Natural Competence as Strategic Survival Attributes

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    Charu Tripathi

    2017-07-01

    Full Text Available Thermophilic environments represent an interesting niche. Among thermophiles, the genus Thermus is among the most studied genera. In this study, we have sequenced the genome of Thermus parvatiensis strain RL, a thermophile isolated from Himalayan hot water springs (temperature >96°C using PacBio RSII SMRT technique. The small genome (2.01 Mbp comprises a chromosome (1.87 Mbp and a plasmid (143 Kbp, designated in this study as pTP143. Annotation revealed a high number of repair genes, a squeezed genome but containing highly plastic plasmid with transposases, integrases, mobile elements and hypothetical proteins (44%. We performed a comparative genomic study of the group Thermus with an aim of analysing the phylogenetic relatedness as well as niche specific attributes prevalent among the group. We compared the reference genome RL with 16 Thermus genomes to assess their phylogenetic relationships based on 16S rRNA gene sequences, average nucleotide identity (ANI, conserved marker genes (31 and 400, pan genome and tetranucleotide frequency. The core genome of the analyzed genomes contained 1,177 core genes and many singleton genes were detected in individual genomes, reflecting a conserved core but adaptive pan repertoire. We demonstrated the presence of metagenomic islands (chromosome:5, plasmid:5 by recruiting raw metagenomic data (from the same niche against the genomic replicons of T. parvatiensis. We also dissected the CRISPR loci wide all genomes and found widespread presence of this system across Thermus genomes. Additionally, we performed a comparative analysis of competence loci wide Thermus genomes and found evidence for recent horizontal acquisition of the locus and continued dispersal among members reflecting that natural competence is a beneficial survival trait among Thermus members and its acquisition depicts unending evolution in order to accomplish optimal fitness.

  6. [Protective effect of arctigenin in GK rats combined with hypertension macroangiopathy].

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    Feng, Qin; Sun, Bao-cun; Xia, Wen-kai

    2015-03-01

    To study the protective effect of Arctigenin in goto-kakizaki (GK) rats combined with hypertension macroangiopathy. Six-week-old GK rats were divided randomly according to blood glucose level into four groups: the model group and low, middle and high dose arctigenin groups (12.5, 25, 50 mg x kg(-1)), with Wistar rats as the normal group. All of GK rats were given high-glucose and high-fat diet. After 16 weeks, GK rats were orally administrated with 10 mg x kg(-1) x d(-1) N-Ω-nitro-L-arginine methyl ester for eight weeks. During the modeling, all of arctigenin groups were orally administrated with different dose of arctigenin twice a day; The model group and the normal group were given solvents. At the beginning, mid-term and end of the experiment, blood glucose was measured. At the end of the experiment, efforts were made to detect blood pressure, collect abdominal aortic blood after anesthesia, fix thoracic aorta after bloodletting to make paraffin sections, observe morphological characteristics and detect the expression of VEGF by immunohistochemistry. According to the results, the blood glucose rose in all GK rats, with no significant difference between the drug group and the model group. At the end of the experiment, the blood pressure significantly increased in GK rats, indicating that Arctigenin could notably reduce the blood pressure in GK rats in a dose-dependent manner. The blood routine test showed increases in both the total white blood cell count and differential blood count, MPV and PDW, abnormal blood platelet parameters and decrease in PLT in GK rats, suggesting that Arctigenin could remarkably reduce the total white blood cell count and differential blood count, MPV and PDW. The thoracic aortic morphological observation revealed obvious endangium lesions in GK rats, demonstrating that Arctigenin could ameliorate the lesion extent. VEGF immumohistochemical staining showed a higher VEGF expression in the model group but lower expression in Arctigenin

  7. Comparative genomics of Thermus thermophilus and Deinococcus radiodurans: divergent routes of adaptation to thermophily and radiation resistance

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    Daly Michael J

    2005-10-01

    Full Text Available Abstract Background Thermus thermophilus and Deinococcus radiodurans belong to a distinct bacterial clade but have remarkably different phenotypes. T. thermophilus is a thermophile, which is relatively sensitive to ionizing radiation and desiccation, whereas D. radiodurans is a mesophile, which is highly radiation- and desiccation-resistant. Here we present an in-depth comparison of the genomes of these two related but differently adapted bacteria. Results By reconstructing the evolution of Thermus and Deinococcus after the divergence from their common ancestor, we demonstrate a high level of post-divergence gene flux in both lineages. Various aspects of the adaptation to high temperature in Thermus can be attributed to horizontal gene transfer from archaea and thermophilic bacteria; many of the horizontally transferred genes are located on the single megaplasmid of Thermus. In addition, the Thermus lineage has lost a set of genes that are still present in Deinococcus and many other mesophilic bacteria but are not common among thermophiles. By contrast, Deinococcus seems to have acquired numerous genes related to stress response systems from various bacteria. A comparison of the distribution of orthologous genes among the four partitions of the Deinococcus genome and the two partitions of the Thermus genome reveals homology between the Thermus megaplasmid (pTT27 and Deinococcus megaplasmid (DR177. Conclusion After the radiation from their common ancestor, the Thermus and Deinococcus lineages have taken divergent paths toward their distinct lifestyles. In addition to extensive gene loss, Thermus seems to have acquired numerous genes from thermophiles, which likely was the decisive contribution to its thermophilic adaptation. By contrast, Deinococcus lost few genes but seems to have acquired many bacterial genes that apparently enhanced its ability to survive different kinds of environmental stresses. Notwithstanding the accumulation of

  8. Phylogenetic analysis of several Thermus strains from Rehai of Tengchong, Yunnan, China.

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    Lin, Lianbing; Zhang, Jie; Wei, Yunlin; Chen, Chaoyin; Peng, Qian

    2005-10-01

    Several Thermus strains were isolated from 10 hot springs of the Rehai geothermal area in Tengchong, Yunnan province. The diversity of Thermus strains was examined by sequencing the 16S rRNA genes and comparing their sequences. Phylogenetic analysis showed that the 16S rDNA sequences from the Rehai geothermal isolates form four branches in the phylogenetic tree and had greater than 95.9% similarity in the phylogroup. Secondary structure comparison also indicated that the 16S rRNA from the Rehai geothermal isolates have unique secondary structure characteristics in helix 6, helix 9, and helix 10 (reference to Escherichia coli). This research is the first attempt to reveal the diversity of Thermus strains that are distributed in the Rehai geothermal area.

  9. Properties of Thermus ruber Strains Isolated from Icelandic Hot Springs and DNA:DNA Homology of Thermus ruber and Thermus aquaticus

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    Sharp, Richard J.; Williams, Ralph A. D.

    1988-01-01

    Seventeen pink-pigmented strains of the genus Thermus were isolated from samples collected from thermal areas of Iceland. The strains were examined by using phenotypic characterization and DNA:DNA homology and were compared with recognized strains. Visually, the strains could be divided into three groups based on their pigmentation; however, spectroscopic studies of the pigments indicated little difference among them. Most strains required a vitamin supplement for growth and used fructose, maltose, mannose, or sucrose as the sole carbon source. In the presence of nitrate, two strains were able to grow under anaerobic conditions. The optimum growth temperature was 60°C; growth did not occur at 30 or 70°C. PMID:16347714

  10. Characterization of the cGMP-dependent protein kinase SmcGK1 of Schistosoma mansoni

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    Silke Leutner

    2011-06-01

    Full Text Available Schistosomes are trematode parasites and of worldwide medical importance for humans and animals. Growth and development of these parasites require a specific host environment, but also permanent communication processes between the two genders. Accumulating molecular evidence indicates that the responsible interactions are mediated by signal transduction processes. Conserved signaling molecules were identified, and first approaches made for their characterization. However, no representative of the conserved family of cGMP-dependent protein kinases (cGKs has been described in this parasite yet. Within the Schistosoma mansoni genome data-set we identified cGK homologs, of which one was investigated in more detail in this study. We present the cloning of SmcGK1, whose sequence shows homology to cGKs of higher eukaryotes. SmcGK1 was found to be gender-independently transcribed in adult schistosomes. The occurrence of SmcGK1 sense and antisense transcripts suggests that the expression of this gene is controlled at the post-transcriptional level. In situ hybridization experiments demonstrated a gonad-preferential expression profile in both genders indicating a role of SmcGK1, at least during sexual development of schistosomes. Using a cGK-specific inhibitor to treat adult schistosomes in vitro finally resulted in a multifaceted phenotype including slow motion, oocyte congestion, and reduced egg production.Esquistossomos são parasitas trematodos de importância médica em todo o mundo para o homem e os animais. O crescimento e o desenvolvimento destes parasitas requerem um ambiente específico do hospedeiro, mas também um processo de comunicação permanente entre parasitas dos dois sexos. Evidência molecular tem se acumulado e indica que as interações são mediadas por processos de transdução de sinal. Moléculas sinalizadoras conservadas foram identificadas, e as primeiras abordagens têm sido feitas para sua caracterização. Contudo, não foi

  11. Thermus caliditerrae sp. nov., a novel thermophilic species isolated from a geothermal area.

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    Ming, Hong; Yin, Yi-Rui; Li, Shuai; Nie, Guo-Xing; Yu, Tian-Tian; Zhou, En-Min; Liu, Lan; Dong, Lei; Li, Wen-Jun

    2014-02-01

    Two thermophilic bacterial strains, designated YIM 77925(T) and YIM 77777, were isolated from two hot springs, one in the Hydrothermal Explosion (Shuirebaozhaqu) area and Frog Mouth Spring in Tengchong county, Yunnan province, south-western China. The taxonomic positions of the two isolates were investigated by a polyphasic approach. Cells of the two strains were Gram-stain-negative, aerobic and rod-shaped. They were able to grow at 50-70 °C, pH 6.0-8.0 and with a NaCl tolerance up to 0.5% (w/v). Colonies are circular, convex, non-transparent and produce yellow pigment. Phylogenetic analyses based on 16S rRNA gene sequences comparison clearly demonstrated that strains YIM 77925(T) and YIM 77777 represent members of the genus Thermus, and they also detected low-level similarities of 16S rRNA gene sequences (below 97%) compared with all other species in this genus. Their predominant menaquinone was MK-8. The genomic DNA G+C contents of strains YIM 77925(T) and YIM 77777 were 65.6 mol% and 67.2 mol%, respectively. Based on the results of physiological and biochemical tests and phylogenetic analyses, strains YIM 77925(T) and YIM 77777 could not be classified as representing any species of the genus Thermus with a validly published name. Thus the two strains are considered to represent a novel species of the genus Thermus, for which the name Thermus caliditerrae sp. nov. is proposed. The type strain is YIM 77925(T) ( = DSM 25901(T) = CCTCC 2012061(T)).

  12. Rhamnolipid-producing thermophilic bacteria of species Thermus and Meiothermus

    Czech Academy of Sciences Publication Activity Database

    Řezanka, Tomáš; Siřišťová, L.; Sigler, Karel

    2011-01-01

    Roč. 15, č. 6 (2011), s. 697-709 ISSN 1431-0651 R&D Projects: GA ČR(CZ) GAP503/11/0215 Institutional research plan: CEZ:AV0Z50200510 Keywords : Thermus sp * T. aquaticus * Meiothermus ruber Subject RIV: EE - Microbiology, Virology Impact factor: 2.941, year: 2011

  13. Bovine herpesvirus type-1 glycoprotein K (gK) interacts with UL20 and is required for infectious virus production

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    Haque, Muzammel; Stanfield, Brent; Kousoulas, Konstantin G.

    2016-12-15

    We have previously shown that the HSV-1 gK and UL20 proteins interact and function in virion envelopment, membrane fusion, and neuronal entry. Alignment of the predicted secondary structures of gKs encoded by BoHV-1, HSV-1, HSV-2, EHV-1 and VZV indicated a high degree of domain conservation. Two BoHV-1 gK-null mutant viruses were created by either gK gene deletion or stop codon insertion. In addition, a V5 epitope-tag was inserted at the carboxyl terminus of gK gene to detect gK. The engineered gK-null mutant viruses failed to replicate and produce viral plaques. Co-immunoprecipitation of gK and UL20 expressed via different methods revealed that gK and UL20 physically interacted in the presence or absence of other viral proteins. Confocal microscopy showed that gK and UL20 colocalized in infected cells. These results indicate that BoHV-1 gK and UL20 may function in a similar manner to other alphaherpesvirus orthologues specified by HSV-1, PRV and EHV-1. -- Highlights: •Glycoprotein K(gK) is conserved among alphaherpesviruses and serves similar functions. •The bovine herpesvirus-1 gK and UL20 proteins physically interact in a similar manner to herpes simplex virus type 1 and equine herpesvirus-1. •The bovine herpesvirus-1 (BoHV-1) gK interacts with UL20 and is essential for virus replication and spread.

  14. Bovine herpesvirus type-1 glycoprotein K (gK) interacts with UL20 and is required for infectious virus production

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    Haque, Muzammel; Stanfield, Brent; Kousoulas, Konstantin G.

    2016-01-01

    We have previously shown that the HSV-1 gK and UL20 proteins interact and function in virion envelopment, membrane fusion, and neuronal entry. Alignment of the predicted secondary structures of gKs encoded by BoHV-1, HSV-1, HSV-2, EHV-1 and VZV indicated a high degree of domain conservation. Two BoHV-1 gK-null mutant viruses were created by either gK gene deletion or stop codon insertion. In addition, a V5 epitope-tag was inserted at the carboxyl terminus of gK gene to detect gK. The engineered gK-null mutant viruses failed to replicate and produce viral plaques. Co-immunoprecipitation of gK and UL20 expressed via different methods revealed that gK and UL20 physically interacted in the presence or absence of other viral proteins. Confocal microscopy showed that gK and UL20 colocalized in infected cells. These results indicate that BoHV-1 gK and UL20 may function in a similar manner to other alphaherpesvirus orthologues specified by HSV-1, PRV and EHV-1. -- Highlights: •Glycoprotein K(gK) is conserved among alphaherpesviruses and serves similar functions. •The bovine herpesvirus-1 gK and UL20 proteins physically interact in a similar manner to herpes simplex virus type 1 and equine herpesvirus-1. •The bovine herpesvirus-1 (BoHV-1) gK interacts with UL20 and is essential for virus replication and spread.

  15. A Metabonomic Comparison of Urinary Changes in Zucker and GK Rats

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    Liang-Cai Zhao

    2010-01-01

    Full Text Available To further investigate pathogenesis and pathogenic process of type 2 diabetes mellitus (T2DM, we compared the urinary metabolic profiling of Zucker obese and Goto-kakizaki (GK rats by NMR-based metabonomics. Principal component analysis (PCA on urine samples of both models rats indicates markedly elevated levels of creatine/creatinine, dimethylamine, and acetoacetate, with concomitantly declined levels of citrate, 2-ketoglurarate, lactate, hippurate, and succinate compared with control rats, respectively. Simultaneously, compared with Zucker obese rats, the GK rats show decreased levels of trimethylamine, acetate, and choline, as well as increased levels of creatine/creatinine, acetoacetate, alanine, citrate, 2-ketoglutarate, succinate, lactate, and hippurate. This study demonstrates metabolic similarities between the two stages of T2DM, including reduced tricarboxylic acid (TCA cycle and increased ketone bodies production. In addition, compared with Zucker obese rats, the GK rats have enhanced concentration of energy metabolites, which indicates energy metabolic changes produced in hyperglycemia stage more than in insulin resistance stage.

  16. Properties and crystal structure of methylenetetrahydrofolate reductase from Thermus thermophilus HB8.

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    Sayaka Igari

    Full Text Available Methylenetetrahydrofolate reductase (MTHFR is one of the enzymes involved in homocysteine metabolism. Despite considerable genetic and clinical attention, the reaction mechanism and regulation of this enzyme are not fully understood because of difficult production and poor stability. While recombinant enzymes from thermophilic organisms are often stable and easy to prepare, properties of thermostable MTHFRs have not yet been reported.MTHFR from Thermus thermophilus HB8, a homologue of Escherichia coli MetF, has been expressed in E. coli and purified. The purified MTHFR was chiefly obtained as a heterodimer of apo- and holo-subunits, that is, one flavin adenine dinucleotide (FAD prosthetic group bound per dimer. The crystal structure of the holo-subunit was quite similar to the β(8α(8 barrel of E. coli MTHFR, while that of the apo-subunit was a previously unobserved closed form. In addition, the intersubunit interface of the dimer in the crystals was different from any of the subunit interfaces of the tetramer of E. coli MTHFR. Free FAD could be incorporated into the apo-subunit of the purified Thermus enzyme after purification, forming a homodimer of holo-subunits. Comparison of the crystal structures of the heterodimer and the homodimer revealed different intersubunit interfaces, indicating a large conformational change upon FAD binding. Most of the biochemical properties of the heterodimer and the homodimer were the same, except that the homodimer showed ≈50% activity per FAD-bound subunit in folate-dependent reactions.The different intersubunit interfaces and rearrangement of subunits of Thermus MTHFR may be related to human enzyme properties, such as the allosteric regulation by S-adenosylmethionine and the enhanced instability of the Ala222Val mutant upon loss of FAD. Whereas E. coli MTHFR was the only structural model for human MTHFR to date, our findings suggest that Thermus MTHFR will be another useful model for this important enzyme.

  17. Thermus tengchongensis sp. nov., isolated from a geothermally heated soil sample in Tengchong, Yunnan, south-west China.

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    Yu, Tian-Tian; Yao, Ji-Cheng; Ming, Hong; Yin, Yi-Rui; Zhou, En-Min; Liu, Min-Jiao; Tang, Shu-Kun; Li, Wen-Jun

    2013-03-01

    A Gram-stain negative aerobic bacterium, designated YIM 77924(T), was isolated from a geothermally heated soil sample collected at Rehai National Park, Tengchong, Yunnan province, south-west China. Growth was found to occur from 55 to 75 °C (optimum 65 °C), pH 6.0-8.0 (optimum pH 7.0) and 0-1 % NaCl (w/v). Cells were observed to be rod-shaped and the colonies convex, circular, smooth, yellow and non-transparent. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain YIM 77924(T) belongs to the genus Thermus. The 16S rRNA gene sequence similarity values between strain YIM 77924(T) and other species of the genus Thermus were all below 97 %. The polar lipids of strain YIM 77924(T) were determined to be aminophospholipid, phospholipid and glycolipid. The predominant respiratory quinone was determined to be MK-8 and the G+C content was 66.64 mol%. The major fatty acids identified were iso-C(16:0), iso-C(15:0), iso-C(17:0) and C(16:0). On the basis of the morphological and chemotaxonomic characteristics as well as genotypic data, strain YIM 77924(T) is proposed to represent a novel species, Thermus tengchongensis sp. nov., in the genus Thermus. The type strain is YIM 77924(T) (=KCTC 32025(T) = CCTCC AB2012063(T)).

  18. Estrogen deprivation aggravates cardiac hypertrophy in nonobese Type 2 diabetic Goto-Kakizaki (GK) rats.

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    Apaijai, Nattayaporn; Charoenphandhu, Narattaphol; Ittichaichareon, Jitjiroj; Suntornsaratoon, Panan; Krishnamra, Nateetip; Aeimlapa, Ratchaneevan; Chattipakorn, Siriporn C; Chattipakorn, Nipon

    2017-10-31

    Both Type 2 diabetes mellitus (T2DM) and estrogen deprivation have been shown to be associated with the development of cardiovascular disease and adverse cardiac remodeling. However, the role of estrogen deprivation on adverse cardiac remodeling in nonobese T2DM rats has not been clearly elucidated. We hypothesized that estrogen-deprivation aggravates adverse cardiac remodeling in Goto-Kakizaki (GK) rats. Wild-type (WT) and GK rats at the age of 9 months old were divided into two subgroups to have either a sham operation (WTS, GKS) or a bilateral ovariectomy (WTO, GKO) ( n = 6/subgroup). Four months after the operation, the rats were killed, and the heart was excised rapidly. Metabolic parameters, cardiomyocytes hypertrophy, cardiac fibrosis, and biochemical parameters were determined. GK rats had hyperglycemia with hypoinsulinemia, and estrogen deprivation did not increase the severity of T2DM. Cardiac hypertrophy, cardiac oxidative stress, and phosphor-antinuclear factor κB were higher in WTO and GKS rats than WTS rats, and they markedly increased in GKO rats compared with GKS rats. Furthermore, cardiac fibrosis, transforming growth factor-β, Bax, phosphor-p38, and peroxisome proliferator- activated receptor γ coactivator-1α expression were increased in GKS and GKO rats compared with the lean rats. However, mitochondrial dynamics proteins including dynamin-related protein 1 and mitofusin-2 were not altered by T2DM and estrogen deprivation. Although estrogen deprivation did not aggravate T2DM in GK rats, it increased the severity of cardiac hypertrophy by provoking cardiac inflammation and oxidative stress in nonobese GK rats. © 2017 The Author(s).

  19. Reciprocal Education Experiences In Two GK-12 Programs: Teachers Learning And Students Teaching In Diverse Settings

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    Mayo, M.; Williams, C.; Rodriguez, T.; Greely, T.; Pyrtle, A. J.; Rivera-Rentas, A. L.; Vilches, M.

    2004-12-01

    The National Science Foundation's Graduate Teaching Fellows in K-12 Education (GK-12) Program has enabled science, technology, engineering and mathematics (STEM) graduate schools across the country to become more active in local area K-12 schools. An overview of a graduate student's experiences, insights gained and lessons learned as a Fellow in the 2003-2004 Universidad Metropolitana's (UMET) environmental science and the 2004-2005 University of South Florida's (USF) ocean science GK-12 Programs is presented. The major goals of the 2003-2004 UMET GK-12 Program were 1) to enrich environmental science teaching and learning via a thematic approach in eight local public schools and 2) to provide UMET graduate students with exposure to teaching methodologies and practical teaching experience. Utilizing examples from local environments in and nearby Carolina, Puerto Rico to teach key science principles at Escuela de la Comunidad Juana Rodriguez Mundo provided numerous opportunities to relate science topics to students' daily life experiences. By 2004, the UMET GK-12 Program had successfully engaged the entire student body (primarily comprised of bilingual minority kindergarten to sixth graders), teachers and school administrators in environment-focused teaching and learning activities. Examples of such activities include tree planting projects to minimize local erosion, conducting a science fair for the first time in many years, and numerous opportunities to experience what "real scientists do" while conducting environmental science investigations. During the 2004-2005 academic year, skills, insights and lessons learned as a UMET GK-12 Fellow are being further enhanced through participation in the USF GK-12 OCEANS Program. The overall objectives of the 2004-2005 USF GK-12 OCEANS assignment at Madeira Beach Elementary School in Saint Petersburg, Florida are to 1) engage students from various ethnic backgrounds and cultures in hands-on science activities, 2) enhance the

  20. NSF GK-12 Fellows as Mentors for K-12 Teachers Participating in Field Research Experiences

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    Ellins, K.; Perry, E.

    2005-12-01

    The University of Texas Institute for Geophysics (UTIG) recognizes the value of providing educational opportunities to K-12 teachers who play a critical role in shaping the minds of young people who are the future of our science. To that end, UTIG established the "Texas Teachers in the Field" program in 2000 to formalize the participation of K-12 teachers in field programs that included UTIG scientists. In 2002, "Texas Teachers in the Field" evolved through UTIG's involvement in a University of Texas at Austin GK-12 project led by the Environmental Sciences Institute, which enabled UTIG to partner a subset of GK-12 Fellows with teachers participating in geophysical field programs. During the three years of the GK-12 project, UTIG successfully partnered four GK-12 Fellows with five K-12 teachers. The Fellows served as mentors to the teachers, as liaisons between UTIG scientists leading field programs and teachers and their students, and as resources in science, mathematics, and technology instruction. Specifically, Fellows prepared teachers and their students for the field investigations, supervised the design of individual Teacher Research Experience (TRE) projects, and helped teachers to develop standards-aligned curriculum resources related to the field program for use in their own classrooms, as well as broader distribution. Although all but one TRE occurred during the school year, Texas school districts and principals were willing to release teachers to participate because the experience and destinations were so extraordinary (i.e., a land-based program in Tierra del Fuego, Argentina; and research cruises to the Southeast Caribbean Sea and Hess Deep in the Pacific Ocean) and carried opportunities to work with scientists from around the world. This exceptional collaboration of GK-12 Fellows, K-12 teachers and research scientists enriches K-12 student learning and promotes greater enthusiasm for science. The level of mentoring, preparation and follow-up provided

  1. Crystallization, characterization and preliminary X-ray crystallographic analysis of GK2848, a putative carbonic anhydrase of Geobacillus kaustophilus

    International Nuclear Information System (INIS)

    Ragunathan, Preethi; Raghunath, Gokul; Kuramitsu, Seiki; Yokoyama, Shigeyuki; Kumarevel, Thirumananseri; Ponnuraj, Karthe

    2013-01-01

    The expression, purification, characterization and crystallization of GK2848, a carbonic anhydrase from G. kaustophilus, are described. The crystals diffracted to a resolution of 2.70 Å. GK2848, a hypothetical protein from the thermophilic organism Geobacillus kaustophilus, was cloned and overexpressed in Escherichia coli. The protein was purified to homogeneity using Ni–NTA affinity-column and gel-filtration chromatography. The purified protein was crystallized using the sitting-drop vapour-diffusion method. The crystals diffracted to a resolution of 2.70 Å and belonged to the orthorhombic space group P2 1 2 1 2. GK2848 bears sequence homology to carbonic anhydrases of various bacterial species, indicating that it belongs to the carbonic anhydrase family of proteins. A subsequent carbonic anhydrase activity assay of GK2848 using the Wilbur–Anderson method confirmed its function as a carbonic anhydrase. A preliminary structure solution was obtained by molecular replacement using MOLREP. Mutation and biochemical characterization of the protein are in progress. The structure and functional analysis of GK2848 might provide valuable information on a novel class of carbonic anhydrases, as none of its homologous structures have been characterized

  2. Differential Changes of Aorta and Carotid Vasodilation in Type 2 Diabetic GK and OLETF Rats: Paradoxical Roles of Hyperglycemia and Insulin

    Directory of Open Access Journals (Sweden)

    Mei-Fang Zhong

    2012-01-01

    Full Text Available We investigated large vessel function in lean Goto-Kakizaki diabetic rats (GK and Otsuka Long-Evans Tokushima Fatty diabetic rats (OLETF with possible roles of hyperglycemia/hyperosmolarity and insulin. Both young and old GK showed marked hyperglycemia with normal insulin level and well-preserved endothelium-dependent and endothelium-independent vasodilation in aorta and carotid artery. There were significant elevations in endothelial/inducible nitric oxide synthase (eNOS/iNOS and inducible/constitutive heme oxygenase (HO-1/HO-2 in GK. The endothelium-dependent vasodilation in GK was inhibited partly by NOS blockade and completely by simultaneous blocking of HO and NOS. In contrast, OLETF showed hyperinsulinemia and mild hyperglycemia but significant endothelium dysfunction beginning at early ages with concomitantly reduced eNOS. Insulin injection corrected hyperglycemia in GK but induced endothelium dysfunction and intima hyperplasia. Hyperglycemia/hyperosmolarity in vitro enhanced vessel eNOS/HO. We suggest that hyperinsulinemia plays a role in endothelium dysfunction in obese diabetic OLETF, while hyperglycemia/hyperosmolarity-induced eNOS/HO upregulation participates in the adaptation of endothelium function in lean diabetic GK.

  3. Regenerating 1 and 3b gene expression in the pancreas of type 2 diabetic Goto-Kakizaki (GK rats.

    Directory of Open Access Journals (Sweden)

    Sophie Calderari

    Full Text Available Regenerating (REG proteins are associated with islet development, β-cell damage, diabetes and pancreatitis. Particularly, REG-1 and REG-3-beta are involved in cell growth/survival and/or inflammation and the Reg1 promoter contains interleukin-6 (IL-6-responsive elements. We showed by transcriptome analysis that islets of Goto-Kakizaki (GK rats, a model of spontaneous type 2 diabetes, overexpress Reg1, 3α, 3β and 3γ, vs Wistar islets. Goto-Kakizaki rat islets also exhibit increased cytokine/chemokine expression/release, particularly IL-6. Here we analyzed Reg1 and Reg3β expression and REG-1 immuno-localization in the GK rat pancreas in relationship with inflammation. Isolated pancreatic islets and acinar tissue from male adult Wistar and diabetic GK rats were used for quantitative RT-PCR analysis. REG-1 immunohistochemistry was performed on paraffin sections with a monoclonal anti-rat REG-1 antibody. Islet cytokine/chemokine release was measured after 48 h-culture. Islet macrophage-positive area was quantified on cryostat sections using anti-CD68 and major histocompatibility complex (MHC class II antibodies. Pancreatic exocrine-to-endocrine Reg1 and Reg3β mRNA ratios were markedly increased in Wistar vs GK rats. Conversely, both genes were upregulated in isolated GK rat islets. These findings were unexpected, because Reg genes are expressed in the pancreatic acinar tissue. However, we observed REG-1 protein labeling in acinar peri-ductal tissue close to islets and around large, often disorganized, GK rat islets, which may retain acinar cells due to their irregular shape. These large islets also showed peri-islet macrophage infiltration and increased release of various cytokines/chemokines, particularly IL-6. Thus, IL-6 might potentially trigger acinar REG-1 expression and secretion in the vicinity of large diabetic GK rat islets. This increased acinar REG-1 expression might reflect an adaptive though unsuccessful response to deleterious

  4. Cloning, purification and crystallization of Thermus thermophilus proline dehydrogenase

    International Nuclear Information System (INIS)

    White, Tommi A.; Tanner, John J.

    2005-01-01

    Cloning, purification and crystallization of T. thermophilus proline dehydrogenase is reported. The detergent n-octyl β-d-glucopyranoside was used to reduce polydispersity, which enabled crystallization. Nature recycles l-proline by converting it to l-glutamate. This four-electron oxidation process is catalyzed by the two enzymes: proline dehydrogenase (PRODH) and Δ 1 -pyrroline-5-carboxylate dehydrogenase. This note reports the cloning, purification and crystallization of Thermus thermophilus PRODH, which is the prototype of a newly discovered superfamily of bacterial monofunctional PRODHs. The results presented here include production of a monodisperse protein solution through use of the detergent n-octyl β-d-glucopyranoside and the growth of native crystals that diffracted to 2.3 Å resolution at Advanced Light Source beamline 4.2.2. The space group is P2 1 2 1 2 1 , with unit-cell parameters a = 82.2, b = 89.6, c = 94.3 Å. The asymmetric unit is predicted to contain two protein molecules and 46% solvent. Molecular-replacement trials using a fragment of the PRODH domain of the multifunctional Escherichia coli PutA protein as the search model (24% amino-acid sequence identity) did not produce a satisfactory solution. Therefore, the structure of T. thermophilus PRODH will be determined by multiwavelength anomalous dispersion phasing using a selenomethionyl derivative

  5. Conformational Dynamics of Thermus aquaticus DNA Polymerase I during Catalysis

    OpenAIRE

    Xu, Cuiling; Maxwell, Brian A.; Suo, Zucai

    2014-01-01

    Despite the fact that DNA polymerases have been investigated for many years and are commonly used as tools in a number of molecular biology assays, many details of the kinetic mechanism they use to catalyze DNA synthesis remain unclear. Structural and kinetic studies have characterized a rapid, pre-catalytic open-to-close conformational change of the Finger domain during nucleotide binding for many DNA polymerases including Thermus aquaticus DNA polymerase I (Taq Pol), a thermostable enzyme c...

  6. Effect of the Nerve Growth Factor Mimetic GK-2 on Brain Structural and Functional State in the Early Postresuscitation Period

    Directory of Open Access Journals (Sweden)

    M. Sh. Avrushchenko

    2012-01-01

    Full Text Available Objective: to evaluate the efficacy of the nerve growth factor mimetic GK-2 used to improve the structural and functional state of the brain in the early postresuscitation period. Material and methods. Cardiac arrest was induced in mature male albino rats for 12 minutes, followed by resuscitation. The neurological state of the resuscitated animals was assessed by a scoring scale. On postresuscitation day 7, the density and composition of neuronal populations of Purkinje cells in the lateral cerebellar region and pyramidal neurons in the hippocampal CA1 sector were determined by a differential morphometric analysis. The results were statistically processed using the ANOVA method. Results. The use of GK-2 was found to accelerate neurological recovery in the resuscitated animals. On day 7 after 12-minute cardiac arrest, the resuscitated animals showed neuronal dystrophic changes and death in the neuronal populations highly susceptible to ischemia. It was shown that the systemic administration of the nerve growth factor mimetic GK-2 contributed to a reduction in the magnitude and depth of postresuscitation changes in the cerebellar Purkinje cells and prevented dystrophic changes in the pyramidal cells of the hippocampal CA1 sector. The findings suggest that GK-2 has a neuroprotective effect in the recovery period after total body ischemia. Conclusion. The results of this study indicate the efficiency of the systemic administration of the nerve growth factor mimetic GK-2 in improving the brain structural and functional state in the early postresuscitation period. This determines perspectives for the use of GK-2 to prevent and correct posthypoxic encephalopathies. Key words: the nerve growth factor mimetic GK-2, postresuscitation period, neuronal dystrophic changes and death, neurological status.

  7. The synthetic parasite-derived peptide GK1 increases survival in a preclinical mouse melanoma model.

    Science.gov (United States)

    Pérez-Torres, Armando; Vera-Aguilera, Jesús; Hernaiz-Leonardo, Juan Carlos; Moreno-Aguilera, Eduardo; Monteverde-Suarez, Diego; Vera-Aguilera, Carlos; Estrada-Bárcenas, Daniel

    2013-11-01

    The therapeutic efficacy of a synthetic parasite-derived peptide GK1, an immune response booster, was evaluated in a mouse melanoma model. This melanoma model correlates with human stage IIb melanoma, which is treated with wide surgical excision; a parallel study employing a surgical treatment was carried out as an instructive goal. C57BL/6 mice were injected subcutaneously in the flank with 2×10(5) B16-F10 murine melanoma cells. When the tumors reached 20 mm3, mice were separated into two different groups; the GK1 group, treated weekly with peritumoral injections of GK1 (10 μg/100 μL of sterile saline solution) and the control group, treated weekly with an antiseptic peritumoral injection of 100 μL of sterile saline solution without further intervention. All mice were monitored daily for clinical appearance, tumor size, and survival. Surgical treatment was performed in parallel when the tumor size was 20 mm3 (group A), 500 mm3 (group B), and >500 mm3 (group C). The GK1 peptide effectively increased the mean survival time by 9.05 days, corresponding to an increase of 42.58%, and significantly delayed tumor growth from day 3 to 12 of treatment. In addition, tumor necrosis was significantly increased (pcancers remains to be determined, and surgical removal remains a challenge for any new experimental treatment of melanoma in mouse models.

  8. Cloning, purification and crystallization of Thermus thermophilus proline dehydrogenase

    Energy Technology Data Exchange (ETDEWEB)

    White, Tommi A.; Tanner, John J., E-mail: tannerjj@missouri.edu [Departments of Chemistry and Biochemistry, University of Missouri-Columbia, Columbia, Missouri 65211 (United States)

    2005-08-01

    Cloning, purification and crystallization of T. thermophilus proline dehydrogenase is reported. The detergent n-octyl β-d-glucopyranoside was used to reduce polydispersity, which enabled crystallization. Nature recycles l-proline by converting it to l-glutamate. This four-electron oxidation process is catalyzed by the two enzymes: proline dehydrogenase (PRODH) and Δ{sup 1}-pyrroline-5-carboxylate dehydrogenase. This note reports the cloning, purification and crystallization of Thermus thermophilus PRODH, which is the prototype of a newly discovered superfamily of bacterial monofunctional PRODHs. The results presented here include production of a monodisperse protein solution through use of the detergent n-octyl β-d-glucopyranoside and the growth of native crystals that diffracted to 2.3 Å resolution at Advanced Light Source beamline 4.2.2. The space group is P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 82.2, b = 89.6, c = 94.3 Å. The asymmetric unit is predicted to contain two protein molecules and 46% solvent. Molecular-replacement trials using a fragment of the PRODH domain of the multifunctional Escherichia coli PutA protein as the search model (24% amino-acid sequence identity) did not produce a satisfactory solution. Therefore, the structure of T. thermophilus PRODH will be determined by multiwavelength anomalous dispersion phasing using a selenomethionyl derivative.

  9. Holomorphic two-spheres in the complex Grassmann manifold G(k, n)

    Indian Academy of Sciences (India)

    of Sciences, Beijing 100049, People's Republic of China. 2Department of Mathematics, University of Science and Technology of China, Hefei, ..... author's knowledge is concerned, there is no corresponding example or counterexample. Example. Through direct calculations, one can show that f : S2 −→ G(k, 2k) defined by.

  10. Two-Point Codes for the Generalised GK curve

    DEFF Research Database (Denmark)

    Barelli, Élise; Beelen, Peter; Datta, Mrinmoy

    2017-01-01

    completely cover and in many cases improve on their results, using different techniques, while also supporting any GGK curve. Our method builds on the order bound for AG codes: to enable this, we study certain Weierstrass semigroups. This allows an efficient algorithm for computing our improved bounds. We......We improve previously known lower bounds for the minimum distance of certain two-point AG codes constructed using a Generalized Giulietti–Korchmaros curve (GGK). Castellanos and Tizziotti recently described such bounds for two-point codes coming from the Giulietti–Korchmaros curve (GK). Our results...

  11. Cesium accumulation by bacterium Thermus sp.TibetanG7: hints for biomineralization of cesiumbearing geyserite in hot springs in Tibet, China

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The bacterium Thermus sp. TibetanG7, isolated from hot springs in Tibet, China, was examined for the ability to accumulate cesium from solutions. Environmental conditions were simulated and the effects of pH, K+, Na+ and K+-regimes were then studied to determine the possible role of the bacterium in the formation of cesium-bearing geyserite around these hot springs. In despite of the inhibition of K+ and Na+, the bacterium Thermus sp. TibetanG7 revealed noticeable accumulation of cesium from solutions, with maximum accumulations of 53.49 and 40.41 μmol Cesium/g cell dry weight in Na+ and K+ inhibition experiments, respectively. The accumulation of cesium by this microorganism is rapid, with 40%―50% accumulated within the first 5 min. K+-deficient cells showed a much higher capacity of cesium accumulation compared with K+-sufficient cells. It is evident that the bacteria within the genus thermus play a significant role in the cesium assembly. The formation of cesium-bearing geyserite is also considered.

  12. Novel N-(pyrimidin-4-ylthiazol-2-amine derivatives as dual-action hypoglycemic agents that activate GK and PPARγ

    Directory of Open Access Journals (Sweden)

    Hui-peng Song

    2011-10-01

    Full Text Available A series of novel N-(pyrimidin-4-ylthiazol-2-amine derivatives have been synthesized and evaluated as glucokinase (GK activators. Ethyl 2-(6-(4-(2-hydroxyethylpiperazin-1-yl-2-methylpyrimidin-4-yl-aminothiazole-5-carboxylate was found to be a potent dual-acting hypoglycemic agent activating both GK and PPARγ. When given orally to normal mice, the compound demonstrated significant efficacy in decreasing the glucose level after oral glucose loading.

  13. Engineering the Substrate Specificity of a Thermophilic Penicillin Acylase from Thermus thermophilus

    Science.gov (United States)

    Torres, Leticia L.; Cantero, Ángel; del Valle, Mercedes; Marina, Anabel; López-Gallego, Fernando; Guisán, José M.

    2013-01-01

    A homologue of the Escherichia coli penicillin acylase is encoded in the genomes of several thermophiles, including in different Thermus thermophilus strains. Although the natural substrate of this enzyme is not known, this acylase shows a marked preference for penicillin K over penicillin G. Three-dimensional models were created in which the catalytic residues and the substrate binding pocket were identified. Through rational redesign, residues were replaced to mimic the aromatic binding site of the E. coli penicillin G acylase. A set of enzyme variants containing between one and four amino acid replacements was generated, with altered catalytic properties in the hydrolyses of penicillins K and G. The introduction of a single phenylalanine residue in position α188, α189, or β24 improved the Km for penicillin G between 9- and 12-fold, and the catalytic efficiency of these variants for penicillin G was improved up to 6.6-fold. Structural models, as well as docking analyses, can predict the positioning of penicillins G and K for catalysis and can demonstrate how binding in a productive pose is compromised when more than one bulky phenylalanine residue is introduced into the active site. PMID:23263966

  14. cAMP-secretion coupling is impaired in diabetic GK/Par rat β-cells: a defect counteracted by GLP-1.

    Science.gov (United States)

    Dolz, Manuel; Movassat, Jamileh; Bailbé, Danielle; Le Stunff, Hervé; Giroix, Marie-Hélène; Fradet, Magali; Kergoat, Micheline; Portha, Bernard

    2011-11-01

    cAMP-raising agents with glucagon-like peptide-1 (GLP-1) as the first in class, exhibit multiple actions that are beneficial for the treatment of type 2 diabetic (T2D) patients, including improvement of glucose-induced insulin secretion (GIIS). To gain additional insight into the role of cAMP in the disturbed stimulus-secretion coupling within the diabetic β-cell, we examined more thoroughly the relationship between changes in islet cAMP concentration and insulin release in the GK/Par rat model of T2D. Basal cAMP content in GK/Par islets was significantly higher, whereas their basal insulin release was not significantly different from that of Wistar (W) islets. Even in the presence of IBMX or GLP-1, their insulin release did not significantly change despite further enhanced cAMP accumulation in both cases. The high basal cAMP level most likely reflects an increased cAMP generation in GK/Par compared with W islets since 1) forskolin dose-dependently induced an exaggerated cAMP accumulation; 2) adenylyl cyclase (AC)2, AC3, and G(s)α proteins were overexpressed; 3) IBMX-activated cAMP accumulation was less efficient and PDE-3B and PDE-1C mRNA were decreased. Moreover, the GK/Par insulin release apparatus appears less sensitive to cAMP, since GK/Par islets released less insulin at submaximal cAMP levels and required five times more cAMP to reach a maximal secretion rate no longer different from W. GLP-1 was able to reactivate GK/Par insulin secretion so that GIIS became indistinguishable from that of W. The exaggerated cAMP production is instrumental, since GLP-1-induced GIIS reactivation was lost in the presence the AC blocker 2',5'-dideoxyadenosine. This GLP-1 effect takes place in the absence of any improvement of the [Ca(2+)](i) response and correlates with activation of the cAMP-dependent PKA-dependent pathway.

  15. Enhanced Glucose Tolerance and Pancreatic Beta Cell Function by Low Dose Aspirin in Hyperglycemic Insulin-Resistant Type 2 Diabetic Goto-Kakizaki (GK Rats

    Directory of Open Access Journals (Sweden)

    Layla Amiri

    2015-07-01

    Full Text Available Background/Aim: Type 2 diabetes is the most common metabolic disorder, characterized by insulin resistance and pancreatic islet beta-cell failure. The most common complications associated with type 2 diabetes are hyperinsulinemia, hyperglycemia, hyperlipidemia, increased inflammatory and reduced insulin response. Aspirin (ASA and other non-steroidal anti-inflammatory drugs (NSAIDs have been associated with the prevention of diabetes, obesity and related cardiovascular disorders. Aspirin has been used in many clinical and experimental trials for the prevention of diabetes and associated complications. Methods: In this study, five month old Goto-Kakizaki (GK rats, which showed signs of mild hyperglycemia (fasting blood glucose 80-95 mg/dl vs 55-60 mg/dl Wistar control rats were used. Two subgroups of GK and Wistar control rats were injected intraperitoneally with 100 mg aspirin/kg body weight/ day for 5 weeks. Animals were sacrificed and blood and tissues were collected after performing glucose tolerance (2 h post 2g IP glucose ingestion tests in experimental and control groups. Results: Aspirin caused a moderate decrease in hyperglycemia. However, we observed a significant improvement in glucose tolerance after ASA treatment in GK rats compared to the nondiabetic Wistar rats. Also, the ASA treated GK rats exhibited a significant decrease in insulinemia. ASA treatment also caused a marked reduction in the pro-inflammatory prostaglandin, PGE2, which was significantly higher in GK rats. On the other hand, no significant organ toxicity was observed after ASA treatment at this dose and time period. However, the total cholesterol and lipoprotein levels were significantly increased in GK rats, which decreased after ASA treatment. Immunofluorescence staining for insulin/glucagon secreting pancreatic cells showed improved beta-cell structural and functional integrity in ASA-treated rats which was also confirmed by SDS-PAGE and Western blot analysis

  16. Detecting DNA synthesis of neointimal formation after catheter balloon injury in GK and in Wistar rats: using 5-ethynyl-2'-deoxyuridine

    Directory of Open Access Journals (Sweden)

    Guo Jingsheng

    2012-12-01

    Full Text Available Abstract Background Neointimal formation plays an important role in the pathogenesis of coronary restenosis after percutaneous coronary intervention (PCI, especially in patients with diabetes mellitus. Recently, some studies have shown that 5-ethynyl-2'-deoxyuridine (EdU incorporation can serve as a novel alternative to the 5-bromo-2'-deoxyuridine (BrdU antibody detection method for detection of DNA synthesis in regenerating avian cochlea, chick embryo and the adult nervous system. However, few studies have been performed to assess the suitability of EdU for detecting DNA synthesis in vascular neointima. Methods The carotid artery balloon injury model was established in Goto-Kakizaki (GK and Wistar rats. A Cell-LightTM EdU Kit was used to detect EdU-labeled cell nuclei of common carotid arteries at day 7 after catheter balloon injury. Different methods of injecting EdU were tested. The protein levels of proliferating cell nuclear antigen (PCNA and p-Akt (Ser473, as well as the mRNA levels of PCNA were evaluated by Western blotting and quantitative real-time PCR (qRT-PCR, respectively. Immunohistochemical staining was also employed to visualize PCNA-positive cells. Results At day 7 after catheter balloon injury, far more EdU-positive and PCNA-positive cells were observed in GK rats. When comparing groups that received different EdU doses, it was found that the percentage of EdU-positive cells at a dose of 100 mg/kg body weight was than at doses of 25 mg/kg and 50 mg/kg. The number of positive cells was significantly higher in the repeated injection group compared to the single injection group. Further, after balloon injury DNA synthesis in GK rats was more notable than in Wistar rats. Neointimal formation in GK rats was more obvious than in Wistar rats. The protein levels of PCNA and p-Akt (Ser473 and the mRNA levels of PCNA were increased in injured rats as compared to uninjured rats, and were significantly higher in GK rats than in Wistar rats

  17. First glycoside hydrolase family 2 enzymes from Thermus antranikianii and Thermus brockianus with β-glucosidase activity

    Directory of Open Access Journals (Sweden)

    Carola eSchröder

    2015-06-01

    Full Text Available Two genes tagh2 and tbgh2 coding for enzymes with hydrolytic activity towards esculin were identified from the extreme thermophilic, aerobic bacteria Thermus antranikianii (Ta and T. brockianus (Tb. Shortened conserved domains predicted a membership of the enzymes of glycoside hydrolase (GH family 2. At present, β-galactosidase activity is found frequently in GH family 2 but β-glucosidase activity has not been reported in this family before. The enzymes TaGH2 and TbGH2 preferred hydrolysis of nitrophenol-linked β-D-glucopyranosides with specific activities of 3,966 U/mg and 660 U/mg, respectively. Residual activities of 40 % (TaGH2 and 51 % (TbGH2 towards 4-NP-β-D-galactopyranoside were observed. Furthermore, TaGH2 hydrolyzed cellobiose. TbGH2, however, showed no activity on cellobiose or lactose. The enzymes exhibited highest activity at 95 °C (TaGH2 and 90 °C (TbGH2 at pH 6.5. Both enzymes were extremely thermostable and thermal activation up to 250 % was observed at temperatures between 50 and 60 °C. Accordingly, the first thermoactive glycoside hydrolase family 2 enzymes with β glucosidase activity have been identified and characterized. The hydrolysis of cellobiose is a unique property of TaGH2 when compared to the enzymes of GH family 2.

  18. Synthesis of rare sugars with L-fuculose-1-phosphate aldolase (FucA) from Thermus thermophilus HB8.

    Science.gov (United States)

    Li, Zijie; Cai, Li; Qi, Qingsheng; Styslinger, Thomas J; Zhao, Guohui; Wang, Peng George

    2011-09-01

    We report herein a one-pot four-enzyme approach for the synthesis of the rare sugars d-psicose, d-sorbose, l-tagatose, and l-fructose with aldolase FucA from a thermophilic source (Thermus thermophilus HB8). Importantly, the cheap starting material DL-GP (DL-glycerol 3-phosphate), was used to significantly reduce the synthetic cost. Copyright © 2011 Elsevier Ltd. All rights reserved.

  19. Ileal Transposition Surgery Decreases Fat Mass and Improves Glucose Metabolism in Diabetic GK Rats: Possible Involvement of FGF21

    Directory of Open Access Journals (Sweden)

    Kemin Yan

    2018-03-01

    Full Text Available Objective: Ileal transposition (IT surgery has been reported to improve glucose and lipid metabolism, and fibroblast growth factor 21 (FGF21 is a powerful metabolic regulator. In the present study, we aimed to investigate the effects of IT surgery on metabolism and its possible relationship with the FGF21 signaling pathway in diabetic Goto-Kakizaki (GK rats.Methods: Ten-week-old male GK rats were subjected to IT surgery with translocation of a 10 cm ileal segment to the proximal jejunum (IT group or sham surgery without the ileum transposition (Sham-IT group. Rats in the no surgery group did not receive any surgical intervention. Six weeks later, body weight, fat mass, fasting blood glucose (FBG, and serum levels of FGF21 and leptin were measured. The expression of the FGF21 signaling pathway and white adipose tissue (WAT browning-related genes in the WAT and liver were evaluated by real-time reverse transcription polymerase chain reaction (RT-qPCR and western blot.Results: IT surgery significantly decreased the body weights and FBG levels and increased the insulin sensitivity of GK rats. The total WAT mass of the IT rats showed a 41.5% reduction compared with the Sham-IT rats, and serum levels of FGF21 and leptin of the IT rats decreased by 26.3 and 61.7%, respectively (all P < 0.05. The mRNA levels of fibroblast growth factor receptor 1 (FGFR1 and its co-receptor β klotho (KLB in the perirenal WAT (pWAT of the IT rats were 1.4- and 2.4-fold that of the Sham-IT rats, respectively, and the FGFR1 protein levels were 1.7-fold of the Sham-IT rats (all P < 0.05. In accordance with the pWAT, the protein levels of FGFR1 and KLB in the epididymal WAT (eWAT of the IT rats notably increased to 3.0- and 3.9-fold of the Sham-IT rats (P < 0.05. Furthermore, uncoupling protein 1 (UCP1 protein levels in the eWAT and pWAT of the IT rats also increased to 2.2- and 2.3-fold of the Sham-IT rats (P < 0.05. However, the protein levels of FGFR1 and KLB in the

  20. Ultraviolet spectrophotometry and optical and infrared photometry of the old nova GK Persei

    Science.gov (United States)

    Wu, Chi-Chao; Holm, Albert V.; Panek, Robert J.; Raymond, John C.; Hartmann, Lee W.; Swank, Jean H.

    1989-01-01

    IUE observations in the 1150-3250-A region were obtained of GK Per during the rise, at the maximum, and during the decline of the 2.5-mag optical outburst in 1981. The results support previous predictions for the interaction of an accretion disk with the magnetic field of an accretion star. The luminosity at minimum is found to be 2.3 solar luminosities, with 1/3 of this being attributed to the cool secondary star.

  1. Restitution of defective glucose-stimulated insulin secretion in diabetic GK rat by acetylcholine uncovers paradoxical stimulatory effect of beta-cell muscarinic receptor activation on cAMP production.

    Science.gov (United States)

    Dolz, Manuel; Bailbé, Danielle; Giroix, Marie-Hélène; Calderari, Sophie; Gangnerau, Marie-Noelle; Serradas, Patricia; Rickenbach, Katharina; Irminger, Jean-Claude; Portha, Bernard

    2005-11-01

    Because acetylcholine (ACh) is a recognized potentiator of glucose-stimulated insulin release in the normal beta-cell, we have studied ACh's effect on islets of the Goto-Kakizaki (GK) rat, a spontaneous model of type 2 diabetes. We first verified that ACh was able to restore the insulin secretory glucose competence of the GK beta-cell. Then, we demonstrated that in GK islets 1) ACh elicited a first-phase insulin release at low glucose, whereas it had no effect in Wistar; 2) total phospholipase C activity, ACh-induced inositol phosphate production, and intracellular free calcium concentration ([Ca2+]i) elevation were normal; 3) ACh triggered insulin release, even in the presence of thapsigargin, which induced a reduction of the ACh-induced [Ca2+]i response (suggesting that ACh produces amplification signals that augment the efficacy of elevated [Ca2+]i on GK exocytosis); 4) inhibition of protein kinase C did not affect [Ca2+]i nor the insulin release responses to ACh; and 5) inhibition of cAMP-dependent protein kinases (PKAs), adenylyl cyclases, or cAMP generation, while not affecting the [Ca2+]i response, significantly lowered the insulinotropic response to ACh (at low and high glucose). In conclusion, ACh acts mainly through activation of the cAMP/PKA pathway to potently enhance Ca2+-stimulated insulin release in the GK beta-cell and, in doing so, normalizes its defective glucose responsiveness.

  2. Isolation and partial characterization of carotenoid underproducing and overproducing mutants from an extremely thermophilic Thermus thermophilus HB27

    International Nuclear Information System (INIS)

    Hoshino, T.; Yoshino, Y.; Guevarra, E.D.; Ishida, S.; Hiruta, T.; Fujii, R.; Nakahara, T.

    1994-01-01

    Twenty-two carotenoid underproducing and thirteen overproducing mutants were obtained from Thermus thermophilus HB27. The strain HB27 was found to produce at least seven colored carotenoids, believed to be identical to those produced by Thermus aquaticus YT1. Based on the results of the genetic analyses performed on twelve carotenoid underproducing mutants, they were classified into three groups; groups 1, 2 and 3. No colored carotenoid was extracted from the cells of mutants belonging to groups 2 and 3, although the accumulation of phytoene, a colorless carotenoid, was observed in group 2 strains. Group 1 was subdivided into groups 1-a and 1-b, where 1-a stains produced neither colored carotenoids nor phytoene and 1-b strains produced two polar colored carotenoids. All of the overproducing mutants produced about twelve times as much seven colored carotenoid mixtures as the parental strain. The mutation loci among all the overproducing mutants were very close to one another, possibly in the same gene. Carotenoid overproducing mutants showed an extensive resistance to UV-irradiation and showed poorer growth at higher temperatures. Carotenoid underproducing mutants were slightly more UV-sensitive but they grew almost normally at higher temperatures. These results suggest that carotenoids are secondary metabolites which are not essential for growth of T. thermophilus

  3. Microturbulence as a third dimension in the G-K giant region of the HR diagram

    International Nuclear Information System (INIS)

    Foy, R.

    1978-01-01

    The structure of the HR diagram in the G-K giant region is complex, because of the funneling effect and of the intersection of evolutionary tracks for different stellar masses. Therefore it is impossible to derive the age of a cool giant from its location in the HR diagram alone. To remove this indeterminacy a third dimension is needed, and the author suggests microturbulence as the appropriate observational parameter. (Auth.)

  4. The Development of Geo-KOMPSAT-2A (GK-2A) Convective Initiation Algorithm over the Korea peninsular

    Science.gov (United States)

    Kim, H. S.; Chung, S. R.; Lee, B. I.; Baek, S.; Jeon, E.

    2016-12-01

    The rapid development of convection can bring heavy rainfall that suffers a great deal of damages to society as well as threatens human life. The high accurate forecast of the strong convection is essentially demanded to prevent those disasters from the severe weather. Since a geostationary satellite is the most suitable instrument for monitoring the single cloud's lifecycle from its formation to extinction, it has been attempted to capture the precursor signals of convection clouds by satellite. Keeping pace with the launch of Geo-KOMPSAT-2A (GK-2A) in 2018, we planned to produce convective initiation (CI) defined as the indicator of potential cloud objects to bring heavy precipitation within two hours. The CI algorithm for GK-2A is composed of four stages. The beginning is to subtract mature cloud pixels, a sort of convective cloud mask by visible (VIS) albedo and infrared (IR) brightness temperature thresholds. Then, the remained immature cloud pixels are clustered as a cloud object by watershed techniques. Each clustering object is undergone 'Interest Fields' tests for IR data that reflect cloud microphysical properties at the current and their temporal changes; the cloud depth, updraft strength and production of glaciations. All thresholds of 'Interest fields' were optimized for Korean-type convective clouds. Based on scores from tests, it is decided whether the cloud object would develop as a convective cell or not. Here we show the result of case study in this summer over the Korea peninsular by using Himawari-8 VIS and IR data. Radar echo and data were used for validation. This study suggests that CI products of GK-2A would contribute to enhance accuracy of the very short range forecast over the Korea peninsular.

  5. The effect of high pressure on the intracellular trehalose synthase activity of Thermus aquaticus.

    Science.gov (United States)

    Dong, Yongsheng; Ma, Lei; Duan, Yuanliang

    2016-01-01

    To understand the effect of high pressure on the intracellular trehalose synthase activity, Thermus aquaticus (T. aquaticus) in the logarithmic growth phase was treated with high-pressure air, and its intracellular trehalose synthase (TSase) activity was determined. Our results indicated that pressure is a factor strongly affecting the cell growth. High pressure significantly attenuated the growth rate of T. aquaticus and shortened the duration of stationary phase. However, after 2 h of culture under 1.0 MPa pressure, the activity of intracellular TSase in T. aquaticus reached its maximum value, indicating that pressure can significantly increase the activity of intracellular TSase in T. aquaticus. Thus the present study provides an important guide for the enzymatic production of trehalose.

  6. A semi-supervised method to detect seismic random noise with fuzzy GK clustering

    International Nuclear Information System (INIS)

    Hashemi, Hosein; Javaherian, Abdolrahim; Babuska, Robert

    2008-01-01

    We present a new method to detect random noise in seismic data using fuzzy Gustafson–Kessel (GK) clustering. First, using an adaptive distance norm, a matrix is constructed from the observed seismic amplitudes. The next step is to find centres of ellipsoidal clusters and construct a partition matrix which determines the soft decision boundaries between seismic events and random noise. The GK algorithm updates the cluster centres in order to iteratively minimize the cluster variance. Multiplication of the fuzzy membership function with values of each sample yields new sections; we name them 'clustered sections'. The seismic amplitude values of the clustered sections are given in a way to decrease the level of noise in the original noisy seismic input. In pre-stack data, it is essential to study the clustered sections in a f–k domain; finding the quantitative index for weighting the post-stack data needs a similar approach. Using the knowledge of a human specialist together with the fuzzy unsupervised clustering, the method is a semi-supervised random noise detection. The efficiency of this method is investigated on synthetic and real seismic data for both pre- and post-stack data. The results show a significant improvement of the input noisy sections without harming the important amplitude and phase information of the original data. The procedure for finding the final weights of each clustered section should be carefully done in order to keep almost all the evident seismic amplitudes in the output section. The method interactively uses the knowledge of the seismic specialist in detecting the noise

  7. A THREE-DIMENSIONAL VIEW OF THE REMNANT OF NOVA PERSEI 1901 (GK Per)

    Energy Technology Data Exchange (ETDEWEB)

    Liimets, T.; Verro, K.; Kolka, I. [Tartu Observatory, 61602 Toravere (Estonia); Corradi, R. L. M.; Rodriguez-Gil, P. [Instituto de Astrofisica de Canarias, Via Lactea s/n, E-38205 La Laguna, Santa Cruz de Tenerife (Spain); Santander-Garcia, M. [Observatorio Astronomico Nacional, Ap. de Correos 112, E-28803 Alcala de Henares, Madrid (Spain); Villaver, E. [Departamento de Fisica Teorica, Universidad Autonoma de Madrid, E-28049 Madrid (Spain)

    2012-12-10

    We present a kinematical study of the optical ejecta of GK Per. It is based on proper-motion measurements of 282 knots from {approx}20 images spanning 25 years. Doppler shifts are also computed for 217 knots. The combination of proper motions and radial velocities allows a unique three-dimensional view of the ejecta to be obtained. The main results are as follows: (1) the outflow is a thick shell in which knots expand with a significant range of velocities, mostly between 600 and 1000 km s{sup -1}, (2) kinematical ages indicate that knots have suffered only a modest deceleration since their ejection a century ago, (3) no evidence for anisotropy in the expansion rate is found, (4) velocity vectors are generally aligned along the radial direction, but a symmetric pattern of non-radial velocities is also observed at specific directions, and (5) the total H{alpha}+[N II] flux has been linearly decreasing at a rate of 2.6% per year in the past decade. The eastern nebular side is fading at a slower rate than the western side. Some of the knots were displayed a rapid change of brightness during the 2004-2011 period. Over a longer timescale, a progressive circularization and homogenization of the nebula are taking place; (6) a kinematic distance of 400 {+-} 30 pc is determined. These results raise some problems with the previous interpretations of the evolution of GK Per. In particular, the idea of a strong interaction of the outflow with the surrounding medium in the southwest quadrant is not supported by our data.

  8. Long-term Spot-Coverage Variations of 13 BY Dra G-K Dwarfs

    Science.gov (United States)

    Alekseev, I. Yu.; Kozhevnikova, A. V.

    2018-06-01

    The results of spot-coverage modeling for 13 active G-K dwarf stars based on many-year photometric observations are presented. The results of UBV RI observations of eight stars performed at the Crimean Astrophysical Observatory were used together with data from the literature in this analysis. The spot-coverage parameters for 13 selected BY Dra active red dwarfs have been redetermined to improve the zonal spot-coverage model for the stellar photospheres, which currently allows for the presence of two active longitudes. Time variations of the spot-activity characteristics of these systems were analyzed with the aim of searching for possible cyclic variations. All the stars, with the exception of OU Gem and BE Cet, show fairly strong correlations between variations in the spot latitudes and spot areas, with absolute values of the correlation coefficients, R(, S), ranging from 0.38 to 0.92. For five stars, an anti-correlation between the mean latitude and area of the spots was found ( R(, S) from-0.24 to-0.73). This behavior may reflect the drift of spots toward the equator in the course of their development. Eight stars feature positive correlations, i.e. the spots drift towards the pole as their areas increase. Nine stars demonstrate activity cycles, which are reflected in photometric variations as well as variations of the spot areas and mean latitudes. The periods of the latitude drift of the spots are found for five stars; the magnitudes of the spot-latitude drift rates are lower than the corresponding value for sunspots by a factor of 1.5-3.

  9. Characterization of recombinant glyoxylate reductase from thermophile Thermus thermophilus HB27.

    Science.gov (United States)

    Ogino, Hiroyasu; Nakayama, Hitoshi; China, Hideyasu; Kawata, Takuya; Doukyu, Noriyuki; Yasuda, Masahiro

    2008-01-01

    A glyoxylate reductase gene from the thermophilic bacterium Thermus thermophilus HB27 (TthGR) was cloned and expressed in Escherichia coli cells. The recombinant enzyme was highly purified to homogeneity and characterized. The purified TthGR showed thermostability up to 70 degrees C. In contrast, the maximum reaction condition was relatively mild (45 degrees C and pH 6.7). Although the kcat values against co-enzyme NADH and NADPH were similar, the Km value against co-enzyme NADH was approximately 18 times higher than that against NADPH. TthGR prefers NADPH rather than NADH as an electron donor. These results indicate that a phosphate group of a co-enzyme affects the binding affinity rather than the reaction efficiency, and TthGR demands appropriate amount of phosphate for a high activity. Furthermore, it was found that the half-lives of TthGR in the presence of 25% dimethyl sulfoxide and diethylene glycol were significantly longer than that in the absence of an organic solvent.

  10. Cloning and analysis of a bifunctional methyltransferase/restriction endonuclease TspGWI, the prototype of a Thermus sp. enzyme family

    Directory of Open Access Journals (Sweden)

    Zylicz-Stachula Agnieszka

    2009-05-01

    Full Text Available Abstract Background Restriction-modification systems are a diverse class of enzymes. They are classified into four major types: I, II, III and IV. We have previously proposed the existence of a Thermus sp. enzyme family, which belongs to type II restriction endonucleases (REases, however, it features also some characteristics of types I and III. Members include related thermophilic endonucleases: TspGWI, TaqII, TspDTI, and Tth111II. Results Here we describe cloning, mutagenesis and analysis of the prototype TspGWI enzyme that recognises the 5'-ACGGA-3' site and cleaves 11/9 nt downstream. We cloned, expressed, and mutagenised the tspgwi gene and investigated the properties of its product, the bifunctional TspGWI restriction/modification enzyme. Since TspGWI does not cleave DNA completely, a cloning method was devised, based on amino acid sequencing of internal proteolytic fragments. The deduced amino acid sequence of the enzyme shares significant sequence similarity with another representative of the Thermus sp. family – TaqII. Interestingly, these enzymes recognise similar, yet different sequences in the DNA. Both enzymes cleave DNA at the same distance, but differ in their ability to cleave single sites and in the requirement of S-adenosylmethionine as an allosteric activator for cleavage. Both the restriction endonuclease (REase and methyltransferase (MTase activities of wild type (wt TspGWI (either recombinant or isolated from Thermus sp. are dependent on the presence of divalent cations. Conclusion TspGWI is a bifunctional protein comprising a tandem arrangement of Type I-like domains; particularly noticeable is the central HsdM-like module comprising a helical domain and a highly conserved S-adenosylmethionine-binding/catalytic MTase domain, containing DPAVGTG and NPPY motifs. TspGWI also possesses an N-terminal PD-(D/EXK nuclease domain related to the corresponding domains in HsdR subunits, but lacks the ATP-dependent translocase module

  11. Crystallization and preliminary crystallographic analysis of a putative glucokinase/hexokinase from Thermus thermophilus

    International Nuclear Information System (INIS)

    Nakamura, Tsutomu; Kashima, Yasuhiro; Mine, Shouhei; Oku, Takashi; Uegaki, Koichi

    2011-01-01

    In this study, a putative glucokinase/hexokinase from T. thermophilus was purified and crystallized. Diffraction data were collected and processed to 2.02 Å resolution. Glucokinase/hexokinase catalyzes the phosphorylation of glucose to glucose 6-phosphate, which is the first step of glycolysis. The open reading frame TTHA0299 of the extreme thermophile Thermus thermophilus encodes a putative glucokinase/hexokinase which contains the consensus sequence for proteins from the repressors, open reading frames and sugar kinases family. In this study, the glucokinase/hexokinase from T. thermophilus was purified and crystallized using polyethylene glycol 8000 as a precipitant. Diffraction data were collected and processed to 2.02 Å resolution. The crystal belonged to space group P2 1 , with unit-cell parameters a = 70.93, b = 138.14, c = 75.16 Å, β = 95.41°

  12. Low molecular weight thiols and thioredoxins are important players in Hg(II) resistance in Thermus thermophilus HB27.

    Science.gov (United States)

    Norambuena, J; Wang, Y; Hanson, T; Boyd, J M; Barkay, T

    2017-11-17

    Mercury (Hg), one of the most toxic and widely distributed heavy metals, has a high affinity for thiol groups. Thiol groups reduce and sequester Hg. Therefore, low molecular weight and protein thiols may be important cell components used in Hg resistance. To date, the role of low molecular weight thiols in Hg-detoxification remains understudied. The mercury resistance ( mer ) operon of Thermus thermophilus suggests an evolutionary link between Hg(II) resistance and low molecular weight thiol metabolism. This mer operon encodes for an enzyme involved in methionine biosynthesis, Oah. Challenge with Hg(II) resulted in increased expression of genes involved in the biosynthesis of multiple low molecular weight thiols (cysteine, homocysteine, and bacillithiol), as well as the thioredoxin system. Phenotypic analysis of gene replacement mutants indicated that Oah contributes to Hg resistance under sulfur limiting conditions, and strains lacking bacillithiol and/or thioredoxins are more sensitive to Hg(II) than the wild type. Growth in presence of either a thiol oxidizing agent or a thiol alkylating agent increased sensitivity to Hg(II). Furthermore, exposure to 3 μM Hg(II) consumed all intracellular reduced bacillithiol and cysteine. Database searches indicate that oah2 is present in all Thermus spp. mer operons. The presence of a thiol related gene was also detected in some alphaprotobacterial mer operons, in which a glutathione reductase gene was present, supporting the role of thiols in Hg(II) detoxification. These results have led to a working model in which LMW thiols act as Hg(II) buffering agents while Hg is reduced by MerA. Importance The survival of microorganisms in presence of toxic metals is central to life's sustainability. The affinity of thiol groups to toxic heavy metals drives microbe-metal interactions and modulate metal toxicity. Mercury detoxification ( mer ) genes likely originated early in microbial evolution among geothermal environments. Little is

  13. Crystallization of ribosomes from Thermus thermophilus

    International Nuclear Information System (INIS)

    Karpova, E.A.; Serdyuk, I.N.; Tarkhovskii, Yu.S.; Orlova, E.V.; Borovyagin, V.L.

    1987-01-01

    An understanding of the molecular bases of the process of protein biosynthesis on the ribosome requires a knowledge of its structure with high three-dimensional resolution involving the method of x-ray crystallographic analysis. The authors report on the production of crystals of the 70S ribosomes from a new source - the highly thermophilic bacterium Thermus thermophilus. Ribosomes for crystallization were obtained from Th. thermophilus strain HB8 by two washings in buffer with high ionic strength. The ribosome preparation was investigated for homogeneity by the method of high-speed sedimentation in a buffer containing 15 mM MgCl 2 , 50 mM NH 4 Cl, and 10 MM Tris-HCl, pH 7.5. Analysis showed that the preparation if homogeneous. The same preparation was investigated for intactness of ribosomal RNA by the method of gel electrophoresis in 2.75% acrylamide 0.5% agarose gel in a buffer containing 30 mM Tris, 30 mM NaH 2 PO 4 , 10 mM EDTA, 1-2% SDS, and 6 M urea. Analysis showed that the preparation possesses intact 16S and 23S RNA. The latter did not degrade, at least in a week of exposure of the ribosomes in buffer solution at 5 0 C. The ribosome preparation had no appreciable RNase activity, which was verified by incubating 4.5 micrograms of ribosomes with 3 micrograms of 14 C-labeled 16S rRna (50 0 C, 90 min) in a buffer containing 10 mM MgCl 2 , 100 mM NH 4 Cl, and 10 mM Tris-HCl, pH/sub 20 0 / 7.5. The incubated nonhydrolyzed RNA was precipitated with 5% trichloroacetic acid and applied on a GF/C filter. The radioactivity was determined in a toluene scintillator on an LS-100C counter

  14. Three-dimensional structure of the enzyme dimanganese catalase from thermus thermophilus at 1 A resolution

    International Nuclear Information System (INIS)

    Antonyuk, S.V.; Melik-Adamyan, V.R.; Popov, A.N.; Lamzin, V.S.; Hempstead, P.D.; Harrison, P.M.; Artymyuk, P.J.; Barynin, V.V.

    2000-01-01

    The crystal structures of two forms of the enzyme dimanganese catalase from Thermus Thermophilus (native and inhibited by chloride) were studied by X-ray diffraction analysis at 1.05 and 0.98 A resolution, respectively. The atomic models of the molecules were refined to the R factors 9.8 and 10%, respectively. The three-dimensional molecular structures are characterized in detail. The analysis of electron-density distributions in the active centers of the native and inhibited enzyme forms revealed that the most flexible side chains of the amino acid residues Lys162 and Glu36 exist in two interrelated conformations. This allowed us to obtain the structural data necessary for understanding the mechanism of enzymatic activity of the dimanganese catalase

  15. Thermophilic amylase from Thermus sp. isolation and its potential application for bioethanol production

    Directory of Open Access Journals (Sweden)

    Amin Fatoni

    2012-11-01

    Full Text Available Limited reserves of fossil energy stimulate researchers to explore for a new alternative energy, such as bioethanol.A thermophilic amylase producing bacterium was isolated from local hot-springs and its characteristic and potential applicationfor bioethanol production was determined. The obtained amylase was studied to determine its optimum temperature, pH,enzymatic reaction time, and substrate concentration. Tapioca waste was used as the substrate to find the potential of theamylase for degrading starch into glucose, and then the process was continued by fermentation to produce bioethanol. Theamylase producer bacterium was proposed as genus Thermus sp. The crude amylase that was obtained has the optimumtemperature of 60°C and optimum pH of 8.0, optimum substrate concentration at 10% (w/w and optimum enzymatic reactiontime of 45 minutes. These enzymes convert the starches of waste tapioca at optimum conditions, with the result of 2.9%ethanol produced from raw materials.

  16. High-level expression, secretion, and purification of the thermostable aqualysin I from Thermus aquaticus YT-1 in Pichia pastoris

    DEFF Research Database (Denmark)

    Oledzka, G.; Dabrowski, Slawomir; Kur, J.

    2003-01-01

    Aqualysin I is a heat-stable subtilisin-type serine protease which is secreted into the culture medium by Thermus aquaticus YT-1, an extreme thermophile. We report the high-level expression of an aqualysin I protein using its native signal sequence for secretion in the methylotrophic yeast, Pichia...... to that of the native enzyme. We also explored the possibility of secreting the GAP expressed aqualysin I in P. pastoris by in-frame fusion of the Saccharomyces cerevisiae alpha-factor secretion signal. However, the levels of secreted pro-aqualysin I particles were approximately 10 times lower, possibly...

  17. Reaction rate of 24Mg(p,γ)25Al

    International Nuclear Information System (INIS)

    Powell, D.C.; Iliadis, C.; Champagne, A.E.; Grossmann, C.A.; Hale, S.E.; Hansper, V.Y.; McLean, L.K.

    1999-01-01

    The proton-capture reaction on 24 Mg has been investigated in the bombarding energy range of E p =0.2-1.7 MeV. Resonance properties (strengths, branching ratios and lifetimes) of low-energy resonances have been measured. From the experimental results, accurate proton partial widths, γ-ray partial widths and total widths (Γ p , Γ γ , and Γ) have been deduced. The present experimental information establishes the 24 Mg+p reaction rates over the temperature range T=0.02-2.0 GK with statistical uncertainties of 5% to 21%. Our recommended reaction rates deviate from previous estimates by 18% to 45%. Based on our results, we can rule out the recent suggestion that the total width of the E R =223 keV resonance has a significant influence on the reaction rates. We also discuss several effects that might give rise to systematic uncertainties in the reaction rates. The astrophysical implications for hydrogen burning of 24 Mg at low stellar temperatures are presented

  18. Crystallization and preliminary X-ray diffraction study of recombinant ribokinase from Thermus Species 2.9

    Energy Technology Data Exchange (ETDEWEB)

    Abramchik, Yu. A. [Russian Academy of Sciences, Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry (Russian Federation); Timofeev, V. I., E-mail: tostars@mail.ru [Russian Academy of Sciences, Shubnikov Institute of Crystallography of Federal Scientific Research Centre “Crystallography and Photonics,” (Russian Federation); Muravieva, T. I.; Esipov, R. S., E-mail: espiov@mx.ibch.ru [Russian Academy of Sciences, Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry (Russian Federation); Kuranova, I. P., E-mail: inna@ns.crys.ras.ru [Russian Academy of Sciences, Shubnikov Institute of Crystallography of Federal Scientific Research Centre “Crystallography and Photonics,” (Russian Federation)

    2016-11-15

    Ribokinase from a thermophilic strain of Thermus species 2.9 belonging to the carbohydrate ribokinase family (EC 2.7.1.15) was isolated, purified, and crystallized. The crystallization conditions were found by the vapor-diffusion technique and were then optimized to apply the capillary counter-diffusion technique. The X-ray diffraction data set was collected from the crystals, which were grown by the counter-diffusion technique, at the SPring-8 synchrotron radiation facility to 2.87 Å resolution. The crystals belong to sp. gr. P12{sub 1}1 and have the following unit-cell parameters: a = 81.613 Å, b = 156.132 Å, c = 87.714 Å, α = γ = 90°, β = 103.819°. The X-ray diffraction data set is suitable for determining the three-dimensional structure of the protein by the molecular-replacement method.

  19. AMP-acetyl CoA synthetase from Leishmania donovani: identification and functional analysis of 'PX4GK' motif.

    Science.gov (United States)

    Soumya, Neelagiri; Kumar, I Sravan; Shivaprasad, S; Gorakh, Landage Nitin; Dinesh, Neeradi; Swamy, Kayala Kambagiri; Singh, Sushma

    2015-04-01

    An adenosine monophosphate forming acetyl CoA synthetase (AceCS) which is the key enzyme involved in the conversion of acetate to acetyl CoA has been identified from Leishmania donovani for the first time. Sequence analysis of L. donovani AceCS (LdAceCS) revealed the presence of a 'PX4GK' motif which is highly conserved throughout organisms with higher sequence identity (96%) to lower sequence identity (38%). A ∼ 77 kDa heterologous protein with C-terminal 6X His-tag was expressed in Escherichia coli. Expression of LdAceCS in promastigotes was confirmed by western blot and RT-PCR analysis. Immunolocalization studies revealed that it is a cytosolic protein. We also report the kinetic characterization of recombinant LdAceCS with acetate, adenosine 5'-triphosphate, coenzyme A and propionate as substrates. Site directed mutagenesis of residues in conserved PX4GK motif of LdAceCS was performed to gain insight into its potential role in substrate binding, catalysis and its role in maintaining structural integrity of the protein. P646A, G651A and K652R exhibited more than 90% loss in activity signifying its indispensible role in the enzyme activity. Substitution of other residues in this motif resulted in altered substrate specificity and catalysis. However, none of them had any role in modulation of the secondary structure of the protein except G651A mutant. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Molecular docking and molecular dynamics simulation studies on Thermus thermophilus leucyl-tRNA synthetase complexed with different amino acids and pre-transfer editing substrates

    OpenAIRE

    Rayevsky A. V.; Tukalo M. A.

    2016-01-01

    Aim. To investigate the structural bases for the amino acid selectivity of the Thermus thermophilus leucyl-tRNA synthetase (LeuRSTT) aminoacylation site and to disclose the binding pattern of pre-transfer editing substrates. Methods. Eight amino acids proposed as semi-cognate substrates for aminoacylation and eight aminoacyl-adenylates (formed from AMP and eight amino acids) were prepared in zwitterions form. The protein structure with a co-crystallized substrate in the aminoacylation site [P...

  1. Conformational Dynamics of Thermus aquaticus DNA Polymerase I during Catalysis

    Science.gov (United States)

    Suo, Zucai

    2014-01-01

    Despite the fact that DNA polymerases have been investigated for many years and are commonly used as tools in a number of molecular biology assays, many details of the kinetic mechanism they use to catalyze DNA synthesis remain unclear. Structural and kinetic studies have characterized a rapid, pre-catalytic open-to-close conformational change of the Finger domain during nucleotide binding for many DNA polymerases including Thermus aquaticus DNA polymerase I (Taq Pol), a thermostable enzyme commonly used for DNA amplification in PCR. However, little has been done to characterize the motions of other structural domains of Taq Pol or any other DNA polymerase during catalysis. Here, we used stopped-flow Förster resonance energy transfer (FRET) to investigate the conformational dynamics of all five structural domains of the full-length Taq Pol relative to the DNA substrate during nucleotide binding and incorporation. Our study provides evidence for a rapid conformational change step induced by dNTP binding and a subsequent global conformational transition involving all domains of Taq Pol during catalysis. Additionally, our study shows that the rate of the global transition was greatly increased with the truncated form of Taq Pol lacking the N-terminal domain. Finally, we utilized a mutant of Taq Pol containing a de novo disulfide bond to demonstrate that limiting protein conformational flexibility greatly reduced the polymerization activity of Taq Pol. PMID:24931550

  2. Science and Engineering of the Environment of Los Angeles: A GK-12 Experiment at Developing Science Communications Skills in UCLA's Graduate Program

    Science.gov (United States)

    Moldwin, M. B.; Hogue, T. S.; Nonacs, P.; Shope, R. E.; Daniel, J.

    2008-12-01

    Many science and research skills are taught by osmosis in graduate programs with the expectation that students will develop good communication skills (speaking, writing, and networking) by observing others, attending meetings, and self reflection. A new National Science Foundation Graduate Teaching Fellows in K- 12 Education (GK-12; http://ehrweb.aaas.org/gk12new/) program at UCLA (SEE-LA; http://measure.igpp.ucla.edu/GK12-SEE-LA/overview.html ) attempts to make the development of good communication skills an explicit part of the graduate program of science and engineering students. SEE-LA places the graduate fellows in two pairs of middle and high schools within Los Angeles to act as scientists-in- residence. They are partnered with two master science teachers and spend two-days per week in the classroom. They are not student teachers, or teacher aides, but scientists who contribute their content expertise, excitement and experience with research, and new ideas for classroom activities and lessons that incorporate inquiry science. During the one-year fellowship, the graduate students also attend a year-long Preparing Future Faculty seminar that discusses many skills needed as they begin their academic or research careers. Students are also required to include a brief (two-page) summary of their research that their middle or high school students would be able to understand as part of their published thesis. Having students actively thinking about and communicating their science to a pre-college audience provides important science communication training and helps contribute to science education. University and local pre- college school partnerships provide an excellent opportunity to support the development of graduate student communication skills while also contributing significantly to the dissemination of sound science to K-12 teachers and students.

  3. Site-directed mutation of a laccase from Thermus thermophilus: Effect on the activity profile

    Directory of Open Access Journals (Sweden)

    Liu Xin

    2012-01-01

    Full Text Available A site-directed mutant R453T of a laccase from Thermus thermophilus HB27 (Tth-laccase was constructed in order to investigate the effect on laccase catalytic properties. The mutated gene was cloned and overexpressed in Escherichia coli. Nickel-affinity purification was achieved and followed by copper ion incorporation. The mature mutated enzyme was quantitatively equal to the wild type. A photometric assay based on the oxidation of the substrate 2,2-azino-bis-(3- ethylbenzthiazoline-6-sulfonate (ABTS was employed in comparison with the wild-type Tth-laccase on catalytic properties. The R453T mutant exhibited improvement in substrate affinity and specific activity at room temperature, whereas those parameters were not significantly influenced when the temperature increased up to 65°C or higher. The mutant had better catalytic activity than that of the wild type at acidic pH. Investigated by circular dichroism spectroscopy, the mutant Tth-laccase displayed similar profiles at low and high temperatures.

  4. Thermus Thermophilus as a Model System for the Study of Ribosomal Antibiotic Resistance

    Science.gov (United States)

    Gregory, Steven T.

    2018-03-01

    Ribosomes are the intracellular ribonucleoprotein machines responsible for the translation of mRNA sequence into protein sequence. As an essential cell component, the ribosome is the target of numerous antibiotics that bind to critical functional sites to impair protein synthesis. Mutations causing resistance to antibiotics arise in antibiotic binding sites, and an understanding of the basis of resistance will be an essential component of efforts to develop new antibiotics by rational drug design. We have identified a number of antibiotic-resistance mutations in ribosomal genes of the thermophilic bacterium Thermus thermophilus. This species offers two primary advantages for examining the structural basis of antibiotic-resistance, in particular, its potential for genetic manipulation and the suitability of its ribosomes for analysis by X-ray crystallography. Mutations we have identified in this organism are in many instances identical to those found in other bacterial species, including important pathogens, a result of the extreme conservation of ribosome functional sites. Here I summarize the advantages of this organism as a model system to study antibiotic-resistance mechanisms at the molecular level.

  5. Engineering of DNA polymerase I from Thermus thermophilus using compartmentalized self-replication.

    Science.gov (United States)

    Aye, Seaim Lwin; Fujiwara, Kei; Ueki, Asuka; Doi, Nobuhide

    2018-05-05

    Although compartmentalized self-replication (CSR) and compartmentalized partnered replication (CPR) are powerful tools for directed evolution of proteins and gene circuits, limitations remain in the emulsion PCR process with the wild-type Taq DNA polymerase used so far, including long run times, low amounts of product, and false negative results due to inhibitors. In this study, we developed a high-efficiency mutant of DNA polymerase I from Thermus thermophilus HB27 (Tth pol) suited for CSR and CPR. We modified the wild-type Tth pol by (i) deletion of the N-terminal 5' to 3' exonuclease domain, (ii) fusion with the DNA-binding protein Sso7d, (iii) introduction of four known effective point mutations from other DNA polymerase mutants, and (iv) codon optimization to reduce the GC content. Consequently, we obtained a mutant that provides higher product yields than the conventional Taq pol without decreased fidelity. Next, we performed four rounds of CSR selection with a randomly mutated library of this modified Tth pol and obtained mutants that provide higher product yields in fewer cycles of emulsion PCR than the parent Tth pol as well as the conventional Taq pol. Copyright © 2018 Elsevier Inc. All rights reserved.

  6. The Deinococcus-Thermus phylum and the effect of rRNA composition on phylogenetic tree construction

    Science.gov (United States)

    Weisburg, W. G.; Giovannoni, S. J.; Woese, C. R.

    1989-01-01

    Through comparative analysis of 16S ribosomal RNA sequences, it can be shown that two seemingly dissimilar types of eubacteria Deinococcus and the ubiquitous hot spring organism Thermus are distantly but specifically related to one another. This confirms an earlier report based upon 16S rRNA oligonucleotide cataloging studies (Hensel et al., 1986). Their two lineages form a distinctive grouping within the eubacteria that deserved the taxonomic status of a phylum. The (partial) sequence of T. aquaticus rRNA appears relatively close to those of other thermophilic eubacteria. e.g. Thermotoga maritima and Thermomicrobium roseum. However, this closeness does not reflect a true evolutionary closeness; rather it is due to a "thermophilic convergence", the result of unusually high G+C composition in the rRNAs of thermophilic bacteria. Unless such compositional biases are taken into account, the branching order and root of phylogenetic trees can be incorrectly inferred.

  7. Structure of TTHA1623, a novel metallo-β-lactamase superfamily protein from Thermus thermophilus HB8

    International Nuclear Information System (INIS)

    Yamamura, Akihiro; Okada, Akitoshi; Kameda, Yasuhiro; Ohtsuka, Jun; Nakagawa, Noriko; Ebihara, Akio; Nagata, Koji; Tanokura, Masaru

    2009-01-01

    The crystal structures of TTHA1623 from T. thermophilus HB8 in an iron-bound and a zinc-bound form have been determined to 2.8 and 2.2 Å resolution, respectively. TTHA1623 is a metallo-β-lactamase superfamily protein from the extremely thermophilic bacterium Thermus thermophilus HB8. Homologues of TTHA1623 exist in a wide range of bacteria and archaea and one eukaryote, Giardia lamblia, but their function remains unknown. To analyze the structural properties of TTHA1623, the crystal structures of its iron-bound and zinc-bound forms have been determined to 2.8 and 2.2 Å resolution, respectively. TTHA1623 possesses an αββα-fold similar to that of other metallo-β-lactamase superfamily proteins with glyoxalase II-type metal coordination. However, TTHA1623 exhibits a putative substrate-binding pocket with a unique shape

  8. G+K 1Σ+/sub g/ double-minimum excited state of H2

    International Nuclear Information System (INIS)

    Glover, R.M.; Weinhold, F.

    1977-01-01

    We have obtained a Born--Oppenheimer potential curve for the previously uncharacterized third 1 Σ + /sub g/ state of H 2 , using a correlated 20-term wavefunction of generalized James--Coolidge type. We find this potential curve to have a double-minimum character, with the inner (Rydberg-like) and outer (''ionic'') wells having minima at about 1.99 and 3.30 bohr, respectively, and an intervening maximum at 2.76 bohr. Unlike the extensively studied E+F double-minimum state, the outer well here appears to be the deeper, by some 450 cm -1 in our calculation. The inner and outer minima can apparently be associated with spectral lines that in experimental tables have previously been attributed to distinct G and K electronic states. The appropriate spectroscopic term symbol of this combined state is accordingly G+K 1 Σ + /sub g/ (1ssigma3dsigma+2pπ 2 )

  9. Insight into the stereospecificity of short-chain thermus thermophilus alcohol dehydrogenase showing pro-S hydride transfer and prelog enantioselectivity.

    Science.gov (United States)

    Pennacchio, Angela; Giordano, Assunta; Esposito, Luciana; Langella, Emma; Rossi, Mosè; Raia, Carlo A

    2010-04-01

    The stereochemistry of the hydride transfer in reactions catalyzed by NAD(H)-dependent alcohol dehydrogenase from Thermus thermophilus HB27 was determined by means of (1)H-NMR spectroscopy. The enzyme transfers the pro-S hydrogen of [4R-(2)H]NADH and exhibits Prelog specificity. Enzyme-substrate docking calculations provided structural details about the enantioselectivity of this thermophilic enzyme. These results give additional insights into the diverse active site architectures of the largely versatile short-chain dehydrogenase superfamily enzymes. A feasible protocol for the synthesis of [4R-(2)H]NADH with high yield was also set up by enzymatic oxidation of 2-propanol-d(8) catalyzed by Bacillus stearothermophilus alcohol dehydrogenase.

  10. Exendin-4, but not dipeptidyl peptidase IV inhibition, increases small intestinal mass in GK rats

    DEFF Research Database (Denmark)

    Simonsen, Lotte; Pilgaard, Sofie; Orskov, Cathrine

    2007-01-01

    Long-term treatment with dipeptidyl peptidase IV inhibitors (DPPIV-I) or glucagon-like peptide (GLP)-1 analogs may potentially affect intestinal growth by down- or upregulating the intestinotrophic hormone GLP-2. This study compared the intestinotrophic effects of 12-wk administration of vehicle......, exendin-4 (Ex-4; 5 nmol/kg bid sc), or DPPIV-I (NN-7201, 10 mg/kg qd orally) in GK rats. Some animals were observed additionally for 9 wk after the end of treatment. Both treatments lowered glycated hemoglobin A1c at wk 12 vs. control (Ex-4, -0.8%; DPPIV-I, -0.4%). Body weight was reduced by Ex-4 compared...... with control (361 +/- 4 vs. 399 +/- 5 g; P weight was identical in all groups...

  11. Cloning, expression, purification, crystallization and initial crystallographic analysis of the preprotein translocation ATPase SecA from Thermus thermophilus

    International Nuclear Information System (INIS)

    Vassylyeva, Marina N.; Mori, Hiroyuki; Tsukazaki, Tomoya; Yokoyama, Shigeyuki; Tahirov, Tahir H.; Ito, Koreaki; Vassylyev, Dmitry G.

    2006-01-01

    The SecA ATPase from T. thermophilus was cloned, expressed, purified and crystallized. Complete diffraction data sets were collected for two crystal forms at 2.8 and 3.5 Å resolution, respectively. Determination of the structure is now in progress. The Thermus thermophilus gene encoding the preprotein translocation ATPase SecA was cloned and expressed and the purified protein was crystallized by the hanging-drop vapour-diffusion technique in two different space groups P3 1(2) 21 (a = b = 168.6, c = 149.8 Å) and P6 1(5) 22 (a = b = 130.9, c = 564.6 Å). The crystals, improved by macroseeding, diffracted to beyond 2.8 and 3.5 Å resolution for the trigonal and hexagonal crystal forms, respectively. Structure determination using the multiple isomorphous replacement method is in progress

  12. Peroxiredoxins: A Model for a Self-Assembling Nanoscale System

    Science.gov (United States)

    2014-08-24

    Introduction 24 Chapter Three describes the identification of a potential peroxiredoxin enzyme in the genome of the thermophilic bacterium Thermus...Discussion (TaqPrx) 80 3.9 Summary of results Comparison of the sequences of thermophilic peroxiredoxins showed that the enzyme reported by Logan and...well as that of the other two variants. A second enzyme was also examined, a thermophilic peroxiredoxin from the bacterium Thermus aquaticus. This

  13. Cloning, expression, purification, crystallization and preliminary X-ray analysis of Thermus aquaticus succinyl-CoA synthetase

    International Nuclear Information System (INIS)

    Joyce, Michael A.; Brownie, Edward R.; Hayakawa, Koto; Fraser, Marie E.

    2007-01-01

    Attempts to crystallize succinyl-CoA synthetase from the thermophile T. aquaticus were thwarted by proteolysis of the β-subunit and preferential crystallization of a truncated form. Crystals of the full-length enzyme were grown after the purification protocol was modified to include frequent additions of protease inhibitors. Succinyl-CoA synthetase (SCS) is an enzyme of the citric acid cycle and is thus found in most species. To date, there are no structures available of SCS from a thermophilic organism. To investigate how the enzyme adapts to higher temperatures, SCS from Thermus aquaticus was cloned, overexpressed, purified and crystallized. Attempts to crystallize the enzyme were thwarted by proteolysis of the β-subunit and preferential crystallization of the truncated form. Crystals of full-length SCS were grown after the purification protocol was modified to include frequent additions of protease inhibitors. The resulting crystals, which diffract to 2.35 Å resolution, are of the protein in complex with Mn 2+ -GDP

  14. Effects of Argonaute on Gene Expression in Thermus thermophilus.

    Directory of Open Access Journals (Sweden)

    Daan C Swarts

    Full Text Available Eukaryotic Argonaute proteins mediate RNA-guided RNA interference, allowing both regulation of host gene expression and defense against invading mobile genetic elements. Recently, it has become evident that prokaryotic Argonaute homologs mediate DNA-guided DNA interference, and play a role in host defense. Argonaute of the bacterium Thermus thermophilus (TtAgo targets invading plasmid DNA during and after transformation. Using small interfering DNA guides, TtAgo can cleave single and double stranded DNAs. Although TtAgo additionally has been demonstrated to cleave RNA targets complementary to its DNA guide in vitro, RNA targeting by TtAgo has not been demonstrated in vivo.To investigate if TtAgo also has the potential to control RNA levels, we analyzed RNA-seq data derived from cultures of four T. thermophilus strain HB27 variants: wild type, TtAgo knockout (Δago, and either strain transformed with a plasmid. Additionally we determined the effect of TtAgo on expression of plasmid-encoded RNA and plasmid DNA levels.In the absence of exogenous DNA (plasmid, TtAgo presence or absence had no effect on gene expression levels. When plasmid DNA is present, TtAgo reduces plasmid DNA levels 4-fold, and a corresponding reduction of plasmid gene transcript levels was observed. We therefore conclude that TtAgo interferes with plasmid DNA, but not with plasmid-encoded RNA. Interestingly, TtAgo presence stimulates expression of specific endogenous genes, but only when exogenous plasmid DNA was present. Specifically, the presence of TtAgo directly or indirectly stimulates expression of CRISPR loci and associated genes, some of which are involved in CRISPR adaptation. This suggests that TtAgo-mediated interference with plasmid DNA stimulates CRISPR adaptation.

  15. Plasminogen-independent initiation of the pro-urokinase activation cascade in vivo. Activation of pro-urokinase by glandular kallikrein (mGK-6) in plasminogen-deficient mice

    DEFF Research Database (Denmark)

    List, K; Jensen, Ole Nørregaard; Bugge, T H

    2000-01-01

    )-antitrypsin. We suggest that mouse glandular kallikrein mGK-6 is an activator of pro-uPA in the mouse urinary tract in vivo. Since this kallikrein is expressed in a number of tissues and also occurs in plasma, it can also be considered a candidate for a physiological pro-uPA activator in other locations....

  16. The subcritical mass limit, 2.4 kgU, for the JCO's precipitation tank

    International Nuclear Information System (INIS)

    Komuro, Yuichi

    1999-01-01

    The critical safety on the precipitation tank in JCO Corporation forming a critical accident in September 30, 1999 had to be guaranteed by limiting amount of uranium contained into charging solution. The limited value of uranic mass in the precipitation tank was determined to be 2.4 kg, and from the dissolution tank positioned at upstream of this tank a solution not excess amount of solution to this value was designed to be transferred. In the 2nd Accident Survey Committee, there were found some discussions on a leading method of this value. In order to answer some requirements for this, here was described on outlines on U.S. Nuclear Safety Guide, TID-7016 Rev. 1, leading method of the limited value in 2.4 kg, and safety tolerance. As a result of reinvestigation, as it was confirmed that 2.4 kg in the limited amount contained an sufficient safety tolerance qualitatively and in comparison with already critical data. (G.K.)

  17. Effect of Qufengtongluo Decoction on PI3K/Akt Signaling Pathway in the Kidney of Type 2 Diabetes Mellitus Rat (GK Rat with Diabetic Nephropathy

    Directory of Open Access Journals (Sweden)

    Wei-Jun Huang

    2018-01-01

    Full Text Available Qufengtongluo (QFTL decoction is an effective treatment for diabetic nephropathy (DN. However, the underlying molecular mechanism is still unclear. In this study, we try to investigate whether QFTL decoction acts via inhibiting PI3K/Akt signaling pathway. Twenty-four GK rats were randomly divided into 3 groups: blank group, sham-operated group, and QFTL group. After model establishment, rats in QFTL group were given QFTL decoction by gavage, while the rest were given pure water. During the 8-week intervention, 24 hr urinal protein was measured every 2-3 weeks. After intervention, kidneys were removed for pathological smear, quantitative real-time PCR, and western blotting to detect expression levels of p-PI3K, p-Akt, PTEN, TGF-β, PI3K mRNA, Akt mRNA, PTEN mRNA, and TGF-β mRNA. QFTL group showed a slighter degree of renal fibrosis in Masson and PASM staining and a greater reduction of 24 hr urinal protein than blank group. Compared to blank group, expression levels of p-PI3K, p-Akt, PI3K mRNA, and Akt mRNA were lower in QFTL group, while expression levels of PTEN and PTEN mRNA were higher. Besides, TGF-β was downregulated by QFTL decoction. In conclusion, this study suggests that QFTL decoction might inhibit PI3K/Akt signaling pathway via activating PTEN and inhibiting TGF-β.

  18. Preface to anthropogenic fluvial sedimentation: Centennial celebration of G.K. Gilbert's Hydraulic-Mining Débris in the Sierra Nevada

    Science.gov (United States)

    James, L. Allan; Phillips, Jonathan D.; Lecce, Scott A.

    2017-10-01

    This special issue celebrates the centennial of the publication of G.K. Gilbert's (1917) monograph, Hydraulic-Mining Débris in the Sierra Nevada, U.S. Geological Survey Professional Paper 105 (PP105). Reasons to celebrate PP105 are manifold. It was the last of four classic monographs that Gilbert wrote in a career that spanned five decades. The monograph, PP105, introduced several important concepts and provided an integrated view of watersheds that was uncommon in its day. It also provided an extreme, lucid example of anthropogenic changes and legacy sediment and how to approach such large-scale phenomena from an objective, quantitative basis.

  19. Purification, crystallization and preliminary X-ray diffraction study on pyrimidine nucleoside phosphorylase TTHA1771 from Thermus thermophilus HB8

    International Nuclear Information System (INIS)

    Shimizu, Katsumi; Kunishima, Naoki

    2007-01-01

    The pyrimidine nucleoside phosphorylase TTHA1771 from T. thermophilus HB8 has been overexpressed, purified and crystallized. The crystals diffract X-rays to 1.8 Å resolution using synchrotron radiation. Pyrimidine nucleoside phosphorylase (PYNP) catalyzes the reversible phosphorolysis of pyrimidines in the nucleotide-synthesis salvage pathway. In order to study the structure–thermostability relationship of this enzyme, PYNP from the extreme thermophile Thermus thermophilus HB8 (TTHA1771) has been cloned, overexpressed and purified. The TTHA1771 protein was crystallized at 291 K using the oil-microbatch method with PEG 4000 as a precipitant. A native data set was collected to 1.8 Å resolution using synchrotron radiation. The crystal belongs to the monoclinic space group P2 1 , with unit-cell parameters a = 58.83, b = 76.23, c = 103.86 Å, β = 91.3°

  20. Crystallization and preliminary crystallographic analysis of molybdenum-cofactor biosynthesis protein C from Thermus thermophilus

    International Nuclear Information System (INIS)

    Kanaujia, Shankar Prasad; Ranjani, Chellamuthu Vasuki; Jeyakanthan, Jeyaraman; Baba, Seiki; Chen, Lirong; Liu, Zhi-Jie; Wang, Bi-Cheng; Nishida, Masami; Ebihara, Akio; Shinkai, Akeo; Kuramitsu, Seiki; Shiro, Yoshitsugu; Sekar, Kanagaraj; Yokoyama, Shigeyuki

    2006-01-01

    The molybdenum-cofactor biosynthesis protein C from T. thermophilus has been crystallized in two different space groups, P2 1 and R32; the crystals diffracted to 1.9 and 1.75 Å resolution, respectively. The Gram-negative aerobic eubacterium Thermus thermophilus is an extremely important thermophilic microorganism that was originally isolated from a thermal vent environment in Japan. The molybdenum cofactor in this organism is considered to be an essential component required by enzymes that catalyze diverse key reactions in the global metabolism of carbon, nitrogen and sulfur. The molybdenum-cofactor biosynthesis protein C derived from T. thermophilus was crystallized in two different space groups. Crystals obtained using the first crystallization condition belong to the monoclinic space group P2 1 , with unit-cell parameters a = 64.81, b = 109.84, c = 115.19 Å, β = 104.9°; the crystal diffracted to a resolution of 1.9 Å. The other crystal form belonged to space group R32, with unit-cell parameters a = b = 106.57, c = 59.25 Å, and diffracted to 1.75 Å resolution. Preliminary calculations reveal that the asymmetric unit contains 12 monomers and one monomer for the crystals belonging to space group P2 1 and R32, respectively

  1. Cloning, expression, purification, crystallization and preliminary X-ray crystallographic study of molybdopterin synthase from Thermus thermophilus HB8

    International Nuclear Information System (INIS)

    Kanaujia, Shankar Prasad; Ranjani, Chellamuthu Vasuki; Jeyakanthan, Jeyaraman; Ohmori, Miwa; Agari, Kazuko; Kitamura, Yoshiaki; Baba, Seiki; Ebihara, Akio; Shinkai, Akeo; Kuramitsu, Seiki; Shiro, Yoshitsugu; Sekar, Kanagaraj; Yokoyama, Shigeyuki

    2007-01-01

    The molybdopterin synthase from T. thermophilus HB8 was cloned, expressed, purified and crystallized. The crystals belong to space group P2 1 and diffracted to a resolution of 1.64 Å. Thermus thermophilus is a Gram-negative aerobic thermophilic eubacterium which can grow at temperatures ranging from 323 to 355 K. In addition to their importance in thermostability or adaptation strategies for survival at high temperatures, the thermostable enzymes in thermophilic organisms contribute to a wide range of biotechnological applications. The molybdenum cofactor in all three kingdoms consists of a tricyclic pyranopterin termed molybdopterin that bears the cis-dithiolene group responsible for molybdenum ligation. The crystals of molybdopterin synthase from T. thermophilus HB8 belong to the primitive monoclinic space group P2 1 , with unit-cell parameters a = 33.94, b = 103.32, c = 59.59 Å, β = 101.3°. Preliminary studies and molecular-replacement calculations reveal the presence of three monomers in the asymmetric unit

  2. Excessive islet NO generation in type 2 diabetic GK rats coincides with abnormal hormone secretion and is counteracted by GLP-1.

    Directory of Open Access Journals (Sweden)

    Albert Salehi

    Full Text Available BACKGROUND: A distinctive feature of type 2 diabetes is inability of insulin-secreting beta-cells to properly respond to elevated glucose eventually leading to beta-cell failure. We have hypothesized that an abnormally increased NO production in the pancreatic islets might be an important factor in the pathogenesis of beta-cell dysfunction. PRINCIPAL FINDINGS: We show now that islets of type 2 spontaneous diabetes in GK rats display excessive NO generation associated with abnormal iNOS expression in insulin and glucagon cells, increased ncNOS activity, impaired glucose-stimulated insulin release, glucagon hypersecretion, and impaired glucose-induced glucagon suppression. Pharmacological blockade of islet NO production by the NOS inhibitor N(G-nitro-L-arginine methyl ester (L-NAME greatly improved hormone secretion from GK islets suggesting islet NOS activity being an important target to inactivate for amelioration of islet cell function. The incretin hormone GLP-1, which is used in clinical practice suppressed iNOS and ncNOS expression and activity with almost full restoration of insulin release and partial restoration of glucagon release. GLP-1 suppression of iNOS expression was reversed by PKA inhibition but unaffected by the proteasome inhibitor MG132. Injection of glucose plus GLP-1 in the diabetic rats showed that GLP-1 amplified the insulin response but induced a transient increase and then a poor depression of glucagon. CONCLUSION: The results suggest that abnormally increased NO production within islet cells is a significant player in the pathogenesis of type 2 diabetes being counteracted by GLP-1 through PKA-dependent, nonproteasomal mechanisms.

  3. Interaction of Thermus thermophilus ArsC enzyme and gold nanoparticles naked-eye assays speciation between As(III) and As(V)

    International Nuclear Information System (INIS)

    Politi, Jane; De Stefano, Luca; Spadavecchia, Jolanda; Casale, Sandra; Fiorentino, Gabriella; Antonucci, Immacolata

    2015-01-01

    The thermophilic bacterium Thermus thermophilus HB27 encodes chromosomal arsenate reductase (TtArsC), the enzyme responsible for resistance to the harmful effects of arsenic. We report on adsorption of TtArsC onto gold nanoparticles for naked-eye monitoring of biomolecular interaction between the enzyme and arsenic species. Synthesis of hybrid biological–metallic nanoparticles has been characterized by transmission electron microscopy (TEM), ultraviolet-visible (UV–vis), dynamic light scattering (DLS) and phase modulated infrared reflection absorption (PM-IRRAS) spectroscopies. Molecular interactions have been monitored by UV–vis and Fourier transform-surface plasmon resonance (FT-SPR). Due to the nanoparticles’ aggregation on exposure to metal salts, pentavalent and trivalent arsenic solutions can be clearly distinguished by naked-eye assay, even at 85 μM concentration. Moreover, the assay shows partial selectivity against other heavy metals. (paper)

  4. Evaluation results of the GlobalWatershed GK-12 Fellowship Program - a model for increased science literacy and partnership

    Science.gov (United States)

    Mayer, A. S.; Vye, E.

    2016-12-01

    The Michigan Tech GlobalWatershed GK-12 Fellowship program bridges the gap between K-12 learning institutions and the scientific community with a focus on watershed research. Michigan Tech graduate students (fellows) work in tandem with teachers on the development of relevant hands-on, inquiry based lesson plans and activities based on their doctoral research projects in watershed science. By connecting students and teachers to state of the art academic research in watershed science, teachers are afforded a meaningful way in which to embed scientific research as a component of K-12 curricula, while mentoring fellows on the most pertinent and essential topics for lesson plan development. Fellows fulfill their vital responsibility of communicating their academic research to a broader public while fostering improved teaching and communication skills. A goal of the project is to increase science literacy among students so they may understand, communicate and participate in decisions made at local, regional, and global levels. The project largely works with schools located in Michigan's western Upper Peninsula but also partners with K-12 systems in Sonora, Mexico. While focusing on local and regional issues, the international element of the project helps expand student, teacher, and fellow worldviews and global awareness of watershed issues and creates meaningful partnerships. Lesson plans are available online and teacher workshops are held regularly to disseminate the wealth of information and resources available to the broader public. Evaluation results indicate that fellows' skill and confidence in their ability to communicate science increased as a results of their participation of the program, as well as their desire to communicate science in their future careers. Teachers' confidence in their capacity to present watershed science to their students increased, along with their understanding of how scientific research contributes to understanding of water

  5. Delaware GK-12: Improvement of Science Education in Vocational Technical High Schools Through Collaborative Learning and Coteaching

    Science.gov (United States)

    Madsen, J.; Skalak, K.; Watson, G.; Scantlebury, K.; Allen, D.; Quillen, A.

    2006-12-01

    With funding from the National Science Foundation, the University of Delaware (UD) in partnership with the New Castle County Vocational Technical School District (NCCoVoTech) in Delaware has initiated a GK-12 Program. In each of year this program, nine full time UD graduate students in the sciences, who have completed all or most of their coursework, will be selected to serve as fellows. Participation in the GK-12 program benefits the graduate fellows in many ways. In addition to gaining general insight into current issues of science education, the fellows enhance their experience as scientific researchers by directly improving their ability to effectively communicate complex quantitative and technical knowledge to an audience with multiple and diverse learning needs. In the first year of this project, fellows have been paired with high school science teachers from NCCoVoTech. These pairs, along with the principal investigators (PIs) of this program have formed a learning community that is taking this opportunity to examine and to reflect on current issues in science education while specifically addressing critical needs in teaching science in vocational technical high schools. By participating in summer workshops and follow-up meetings facilitated by the PIs, the fellows have been introduced to a number of innovative teaching strategies including problem-based learning (PBL). Fellow/teacher pairs have begun to develop and teach PBL activities that are in agreement with State of Delaware science standards and that support student learning through inquiry. Fellows also have the opportunity to engage in coteaching with their teacher partner. In this "teaching at the elbow of another", fellows will gain a better understanding of and appreciation for the complexities and nuances of teaching science in vocational technical high schools. While not taught as a stand-alone course in NCCoVoTech high schools, earth science topics are integrated into the science curriculum at

  6. Summary of the GK15 ground‐motion prediction equation for horizontal PGA and 5% damped PSA from shallow crustal continental earthquakes

    Science.gov (United States)

    Graizer, Vladimir;; Kalkan, Erol

    2016-01-01

    We present a revised ground‐motion prediction equation (GMPE) for computing medians and standard deviations of peak ground acceleration (PGA) and 5% damped pseudospectral acceleration (PSA) response ordinates of the horizontal component of randomly oriented ground motions to be used for seismic‐hazard analyses and engineering applications. This GMPE is derived from the expanded Next Generation Attenuation (NGA)‐West 1 database (see Data and Resources; Chiou et al., 2008). The revised model includes an anelastic attenuation term as a function of quality factor (Q0) to capture regional differences in far‐source (beyond 150 km) attenuation, and a new frequency‐dependent sedimentary‐basin scaling term as a function of depth to the 1.5  km/s shear‐wave velocity isosurface to improve ground‐motion predictions at sites located on deep sedimentary basins. The new Graizer–Kalkan 2015 (GK15) model, developed to be simple, is applicable for the western United States and other similar shallow crustal continental regions in active tectonic environments for earthquakes with moment magnitudes (M) 5.0–8.0, distances 0–250 km, average shear‐wave velocities in the upper 30 m (VS30) 200–1300  m/s, and spectral periods (T) 0.01–5 s. Our aleatory variability model captures interevent (between‐event) variability, which decreases with magnitude and increases with distance. The mixed‐effect residuals analysis reveals that the GK15 has no trend with respect to the independent predictor parameters. Compared to our 2007–2009 GMPE, the PGA values are very similar, whereas spectral ordinates predicted are larger at T<0.2  s and they are smaller at longer periods.

  7. Arsenate reductase from Thermus thermophilus conjugated to polyethylene glycol-stabilized gold nanospheres allow trace sensing and speciation of arsenic ions.

    Science.gov (United States)

    Politi, Jane; Spadavecchia, Jolanda; Fiorentino, Gabriella; Antonucci, Immacolata; De Stefano, Luca

    2016-10-01

    Water sources pollution by arsenic ions is a serious environmental problem all around the world. Arsenate reductase enzyme (TtArsC) from Thermus thermophilus extremophile bacterium, naturally binds arsenic ions, As(V) and As (III), in aqueous solutions. In this research, TtArsC enzyme adsorption onto hybrid polyethylene glycol-stabilized gold nanoparticles (AuNPs) was studied at different pH values as an innovative nanobiosystem for metal concentration monitoring. Characterizations were performed by UV/Vis and circular dichroism spectroscopies, TEM images and in terms of surface charge changes. The molecular interaction between arsenic ions and the TtArsC-AuNPs nanobiosystem was also monitored at all pH values considered by UV/Vis spectroscopy. Tests performed revealed high sensitivities and limits of detection equal to 10 ± 3 M -12 and 7.7 ± 0.3 M -12 for As(III) and As(V), respectively. © 2016 The Author(s).

  8. Production of a mouse strain with impaired glucose tolerance by systemic heterozygous knockout of the glucokinase gene and its feasibility as a prediabetes model

    Science.gov (United States)

    SAITO, Mikako; KANEDA, Asako; SUGIYAMA, Tae; IIDA, Ryousuke; OTOKUNI, Keiko; KABURAGI, Misako; MATSUOKA, Hideaki

    2015-01-01

    Exon II of glucokinase (Gk) was deleted to produce a systemic heterozygous Gk knockout (Gk+/−) mouse. The relative expression levels of Gk in the heart, lung, liver, stomach, and pancreas in Gk+/− mice ranged from 0.41–0.68 versus that in wild (Gk+/+) mice. On the other hand, its expression levels in the brain, adipose tissue, and muscle ranged from 0.95–1.03, and its expression levels in the spleen and kidney were nearly zero. Gk knockout caused no remarkable off-target effect on the expression of 7 diabetes causing genes (Shp, Hnf1a, Hnf1b, Irs1, Irs2, Kir6.2, and Pdx1) in 10 organs. The glucose tolerance test was conducted to determine the blood glucose concentrations just after fasting for 24 h (FBG) and at 2 h after high-glucose application (GTT2h). The FBG-GTT2h plots obtained with the wild strain fed the control diet (CD), Gk+/− strain fed the CD, and Gk+/− strain fed the HFD were distributed in separate areas in the FBG-GTT2h diagram. The respective areas could be defined as the normal state, prediabetes state, and diabetes state, respectively. Based on the results, the criteria for prediabetes could be defined for the Gk+/− strain developed in this study. PMID:25765873

  9. Pb2+ Effects on Growth, Lipids, and Protein and DNA Profiles of the Thermophilic Bacterium Thermus Thermophilus

    Directory of Open Access Journals (Sweden)

    Barbara Nicolaus

    2016-12-01

    Full Text Available Extremophiles are organisms able to thrive in extreme environmental conditions and some of them show the ability to survive high doses of heavy metals thanks to defensive mechanisms provided by primary and secondary metabolic products, i.e., extremolytes, lipids, and extremozymes. This is why there is a growing scientific and industrial interest in the use of thermophilic bacteria in a host of tasks, from the environmental detoxification of heavy metal to industrial activities, such as bio-machining and bio-metallurgy. In this work Thermus thermophilus was challenged against increasing Pb2+ concentrations spanning from 0 to 300 ppm in order to ascertain the sensitiveness of this bacteria to the Pb environmental pollution and to give an insight on its heavy metal resistance mechanisms. Analysis of growth parameters, enzyme activities, protein profiles, and lipid membrane modifications were carried out. In addition, genotyping analysis of bacteria grown in the presence of Pb2+, using random amplified polymorphic DNA-PCR and DNA melting evaluation, were also performed. A better knowledge of the response of thermophilic bacteria to the different pollutants, as heavy metals, is necessary for optimizing their use in remediation or decontamination processes.

  10. Crystal structure of a thermostable Old Yellow Enzyme from Thermus scotoductus SA-01

    International Nuclear Information System (INIS)

    Opperman, Diederik J.; Sewell, Bryan T.; Litthauer, Derek; Isupov, Mikhail N.; Littlechild, Jennifer A.; Heerden, Esta van

    2010-01-01

    Recent characterization of the chromate reductase (CrS) from the thermophile Thermus scotoductus SA-01 revealed this enzyme to be related to the Old Yellow Enzyme (OYE) family. Here, we report the structure of a thermostable OYE homolog in its holoform at 2.2 A as well as its complex with p-hydroxybenzaldehyde (pHBA). The enzyme crystallized as octamers with the monomers showing a classical TIM barrel fold which upon dimerization yields the biologically active form of the protein. A sulfate ion is bound above the si-side of the non-covalently bound FMN cofactor in the oxidized solved structure but is displaced upon pHBA binding. The active-site architecture is highly conserved as with other members of this enzyme family. The pHBA in the CrS complex is positioned by hydrogen bonding to the two conserved catalytic-site histidines. The most prominent structural difference between CrS and other OYE homologs is the size of the 'capping domain'. Thermostabilization of the enzyme is achieved in part through increased proline content within loops and turns as well as increased intersubunit interactions through hydrogen bonding and complex salt bridge networks. CrS is able to reduce the C=C bonds of α,β-unsaturated carbonyl compounds with a preference towards cyclic substrates however no activity was observed towards β-substituted substrates. Mutational studies have confirmed the role of Tyr177 as the proposed proton donor although reduction could still occur at a reduced rate when this residue was mutated to phenylalanine.

  11. Identification and characterization of preferred DNA-binding sites for the Thermus thermophilus transcriptional regulator FadR.

    Directory of Open Access Journals (Sweden)

    Minwoo Lee

    Full Text Available One of the primary transcriptional regulators of fatty acid homeostasis in many prokaryotes is the protein FadR. To better understand its biological function in the extreme thermophile Thermus thermophilus HB8, we sought to first determine its preferred DNA-binding sequences in vitro using the combinatorial selection method Restriction Endonuclease Protection, Selection, and Amplification (REPSA and then use this information to bioinformatically identify potential regulated genes. REPSA determined a consensus FadR-binding sequence 5´-TTRNACYNRGTNYAA-3´, which was further characterized using quantitative electrophoretic mobility shift assays. With this information, a search of the T. thermophilus HB8 genome found multiple operons potentially regulated by FadR. Several of these were identified as encoding proteins involved in fatty acid biosynthesis and degradation; however, others were novel and not previously identified as targets of FadR. The role of FadR in regulating these genes was validated by physical and functional methods, as well as comparative genomic approaches to further characterize regulons in related organisms. Taken together, our study demonstrates that a systematic approach involving REPSA, biophysical characterization of protein-DNA binding, and bioinformatics can be used to postulate biological roles for potential transcriptional regulators.

  12. Areca nut extract up-regulates prostaglandin production, cyclooxygenase-2 mRNA and protein expression of human oral keratinocytes.

    Science.gov (United States)

    Jeng, J H; Ho, Y S; Chan, C P; Wang, Y J; Hahn, L J; Lei, D; Hsu, C C; Chang, M C

    2000-07-01

    There are about 600 million betel quid (BQ) chewers in the world. BQ chewing is associated with increased incidence of oral cancer and submucous fibrosis. In this study, areca nut (AN) extract (200-800 microg/ml) induced the prostaglandin E(2) (PGE(2)) production by 1. 4-3.4-fold and 6-keto-PGF(1 alpha) production by 1.1-1.7-fold of gingival keratinocytes (GK), respectively, following 24 h of exposure. Exposure of GK to AN extract (>400 microg/ml) led to cell retraction and intracellular vacuoles formation. At concentrations of 800 and 1200 microg/ml, AN extract induced cell death at 21-24 and 32-52% as detected by MTT assay and cellular lactate dehydrogenase release, respectively. Interestingly, AN-induced morphological changes of GK are reversible. GK can still proliferate following exposure to AN extract. Cytotoxicity of AN extract cannot be inhibited by indomethacin (1 microM) and aspirin (50 microM), indicating that prostaglandin (PG) production is not the major factor responsible for AN cytotoxicity. PGE(2) exhibited little effect on the growth of GK at concentrations ranging from 100-1000 pg/ml. Stimulating GK production of PGs by AN extract could be due to induction of cyclooxygenase-2 (COX-2) mRNA expression and protein production. These results suggest that AN ingredients are critical in the pathogenesis of oral submucous fibrosis and oral cancer via their stimulatory effects on the PGs, COX-2 production and associated tissue inflammatory responses. AN cytotoxicity to GK is not directly mediated by COX-2 stimulation and PG production.

  13. Cloning, sequencing, and expression of dnaK-operon proteins from the thermophilic bacterium Thermus thermophilus.

    Science.gov (United States)

    Osipiuk, J; Joachimiak, A

    1997-09-12

    We propose that the dnaK operon of Thermus thermophilus HB8 is composed of three functionally linked genes: dnaK, grpE, and dnaJ. The dnaK and dnaJ gene products are most closely related to their cyanobacterial homologs. The DnaK protein sequence places T. thermophilus in the plastid Hsp70 subfamily. In contrast, the grpE translated sequence is most similar to GrpE from Clostridium acetobutylicum, a Gram-positive anaerobic bacterium. A single promoter region, with homology to the Escherichia coli consensus promoter sequences recognized by the sigma70 and sigma32 transcription factors, precedes the postulated operon. This promoter is heat-shock inducible. The dnaK mRNA level increased more than 30 times upon 10 min of heat shock (from 70 degrees C to 85 degrees C). A strong transcription terminating sequence was found between the dnaK and grpE genes. The individual genes were cloned into pET expression vectors and the thermophilic proteins were overproduced at high levels in E. coli and purified to homogeneity. The recombinant T. thermophilus DnaK protein was shown to have a weak ATP-hydrolytic activity, with an optimum at 90 degrees C. The ATPase was stimulated by the presence of GrpE and DnaJ. Another open reading frame, coding for ClpB heat-shock protein, was found downstream of the dnaK operon.

  14. Production, purification and characterization of polyclonal antibody against the truncated gK of the duck enteritis virus

    Directory of Open Access Journals (Sweden)

    Zhang Shunchuan

    2010-09-01

    Full Text Available Abstract Duck virus enteritis (DVE is an acute, contagious herpesvirus infection of ducks, geese, and swans, which has produced significant economic losses in domestic and wild waterfowl. With the purpose of decreasing economic losses in the commercial duck industry, studying the unknown glycoprotein K (gK of DEV may be a new method for preferably preventing and curing this disease. So this is the first time to product and purify the rabbit anti-tgK polyclonal antibody. Through the western blot and ELISA assay, the truncated glycoprotein K (tgK has good antigenicity, also the antibody possesses high specificity and affinity. Meanwhile the rabbit anti-tgK polyclonal antibody has the potential to produce subunit vaccines and the functions of neutralizing DEV and anti-DEV infection because of its neutralization titer. Indirect immunofluorescent microscopy using the purified rabbit anti-tgK polyclonal antibody as diagnostic antibody was susceptive to detect a small quantity of antigen in tissues or cells. This approach also provides effective experimental technology for epidemiological investigation and retrospective diagnose of the preservative paraffin blocks.

  15. Dramatic change of the recurrence time and outburst parameters of the intermediate polar GK Persei

    Science.gov (United States)

    Šimon, V.

    2002-02-01

    This analysis has shown that the intermediate polar GK Per experienced a very striking evolution. Its outbursts became wider and brighter in the last five decades. These changes were accompanied by striking variations of the recurrence time T_C, from 385 days within the years 1948-1967 to 890 days in 1970. Nowadays, T_C displays a strong trend of linear increase. Decrease of irradiation of the disk by the WD, combined with the decrease of viscosity, offers a plausible explanation. It is argued that variations of the mass transfer rate are unlikely to play a major role. The morphology of the outburst light curves in the optical and the X-ray region is also studied. The decay branch in the optical remains remarkably similar for all the events while the largest changes of the light curve occur in the rising branch. This can be explained if the thermal instability may start at different distances from the disk center. The quiescent level of brightness does not display any secular trend in recent decades but a wave on the time scale of about 30 years with the full amplitude of 0.3 m_v, probably due to activity of the cool star, is detected. This research has made use of the AFOEV database, operated at CDS, France, and the observations provided by the ASM/RXTE team.

  16. Cross Section Measurements of the Reaction 23Na(p, γ)24Mg

    Science.gov (United States)

    Boeltzig, Axel; Deboer, Richard James; Macon, Kevin; Wiescher, Michael; Best, Andreas; Imbriani, Gianluca; Gyürky, György; Strieder, Frank

    2017-09-01

    The reaction 23Na(p, γ)24Mg can provide a link from the NeNa to the MgAl cycle in stellar burning and is therefore of interest in nuclear astrophysics. To determine the reaction rates at stellar temperatures, new cross section measurements at low proton energies have been performed recently, and further experiments are underway. The current cross section data implies that the reaction rate up to temperatures of 1 GK is determined by a few narrow resonances and direct capture. Complementary to these experimental efforts at low proton energies, cross section measurements at higher energies can help to constrain the direct capture and broad resonance contributions to the cross section and reduce the uncertainty of the extrapolation towards stellar energies. In this paper we report an experiment to measure the 23Na(p, γ)24Mg cross section with a solid target setup at the St. ANA 5U accelerator at the University of Notre Dame. The experiment and the current status of data analysis will be described. This work benefited from support by the National Science Foundation under Grant No. PHY-1430152 (JINA-CEE), the Nuclear Science Laboratory (NSL), the Istituto Nazionale di Fisica Nucleare (INFN), and the Gran Sasso Science Institute (GSSI).

  17. The intermediate polar GK Persei: An unstable relation of the X-ray and the optical intensities in a series of outbursts

    Science.gov (United States)

    Šimon, V.

    2015-03-01

    Context. GK Per is an intermediate polar that has been displaying dwarf nova outbursts since the middle of the twentieth century. Aims: I analyzed a series of such outbursts in the optical and X-ray bands. I pay attention to the relation of intensities of the optical and X-ray emissions, and its reproducibility in a series of these consecutive outbursts. Methods: This analysis uses the data from the BAT/Swift, ASM/RXTE, AAVSO, and AFOEV databases. It investigates the relation of the time evolution of the profiles of outbursts in the individual bands (hard X-ray, medium/hard X-ray, and optical). Results: This analysis shows that the X-ray intensity steeply rises only in the start of the optical outburst and steeply declines only when the optical outburst comes to its end. However, the 1.5-50 keV band intensity saturates and balances on a plateau during the outburst. (The longer the outburst, the longer its plateau.) The peak X-ray intensities of this series display a significantly narrower range than the optical ones (a factor of about two versus a factor of about eight). This implies a discrepancy between the mass flow through the disk and the production of the X-ray emission via bremsstrahlung at the polar caps of the white dwarf. This discrepancy is the largest in the time of the peak optical intensity when the whole disk (or at least its inner part) is in the hot state and the flow of matter through the disk is the greatest. This study shows that a series of outbursts constitutes more general properties of this discrepancy. I argue that the saturation of the X-ray luminosity in outburst cannot be caused by a dominant increase in X-ray absorption. In the interpretation, large structural changes of the accreting regions at the magnetic poles of the white dwarf occur during the outburst. A buried shock proposed by some authors for polars is also promising for explaining the X-ray light curves of outbursts of GK Per. This research made use of the BAT/Swift, ASM

  18. X-RAY FADING AND EXPANSION IN THE “MINIATURE SUPERNOVA REMNANT” OF GK PERSEI

    Energy Technology Data Exchange (ETDEWEB)

    Takei, D. [Institute of Physical and Chemical Research (RIKEN), RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo, Hyogo 679-5148 (Japan); Drake, J. J.; Slane, P. [Smithsonian Astrophysical Observatory, 60 Garden Street, Cambridge, MA 02138 (United States); Yamaguchi, H. [NASA Goddard Space Flight Center, Code 662, Greenbelt, MD 20771 (United States); Uchiyama, Y. [Department of Physics, Rikkyo University, 3-34-1 Nishi-Ikebukuro, Toshima, Tokyo 171-8501 (Japan); Katsuda, S., E-mail: takei@spring8.or.jp [Japan Aerospace Exploration Agency, Institute of Space and Astronautical Science, 3-1-1 Yoshino-dai, Chuo-ku, Sagamihara, Kanagawa 252-5210 (Japan)

    2015-03-10

    We report on a second epoch of Chandra X-ray imaging spectroscopy of the spatially resolved old nova remnant GK Persei. An ACIS-S3 observation of 97.4 ks was conducted in 2013 November after a lapse of 13.8 yr from the last visit in 2000. The X-ray emitting nebula appeared more faint and patchy compared with the first epoch. The flux decline was particularly evident in fainter regions and the mean decline was 30%–40% in the 0.5–1.2 keV energy band. A typical expansion of the brightest part of the remnant was 1.″9, which corresponds to an expansion rate of 0.″14 yr{sup −1}. The soft X-ray spectra extracted from both the 2000 and 2013 data can be explained by a non-equilibrium ionization collisional plasma model convolved with interstellar absorption, though do not allow us to constrain the origin of the flux evolution. The plasma temperature has not significantly evolved since the 2000 epoch and we conclude that the fading of the X-ray emission is due largely to expansion. This implies that recent expansion has been into a lower density medium, a scenario that is qualitatively consistent with the structure of the circumstellar environment photographed soon after the initial explosion more than a century ago. Fainter areas are fading more quickly than brighter areas, indicating that they are fainter because of a lower ambient medium density and consequently more rapid expansion.

  19. Inactivation and unfolding of protein tyrosine phosphatase from Thermus thermophilus HB27 during urea and guanidine hydrochloride denaturation.

    Directory of Open Access Journals (Sweden)

    Yejing Wang

    Full Text Available The effects of urea and guanidine hydrochloride (GdnHCl on the activity, conformation and unfolding process of protein tyrosine phosphatase (PTPase, a thermostable low molecular weight protein from Thermus thermophilus HB27, have been studied. Enzymatic activity assays showed both urea and GdnHCl resulted in the inactivation of PTPase in a concentration and time-dependent manner. Inactivation kinetics analysis suggested that the inactivation of PTPase induced by urea and GdnHCl were both monophasic and reversible processes, and the effects of urea and GdnHCl on PTPase were similar to that of mixed-type reversible inhibitors. Far-ultraviolet (UV circular dichroism (CD, Tryptophan and 1-anilinonaphthalene -8-sulfonic acid (ANS fluorescence spectral analyses indicated the existence of a partially active and an inactive molten globule-like intermediate during the unfolding processes induced by urea and GdnHCl, respectively. Based on the sequence alignment and the homolog Tt1001 protein structure, we discussed the possible conformational transitions of PTPase induced by urea and GdnHCl and compared the conformations of these unfolding intermediates with the transient states in bovine PTPase and its complex structures in detail. Our results may be able to provide some valuable clues to reveal the relationship between the structure and enzymatic activity, and the unfolding pathway and mechanism of PTPase.

  20. Crystal Structure of the 30S Ribosomal Subunit from Thermus Thermophilus: Purification, Crystallization and Structure Determination

    International Nuclear Information System (INIS)

    Clemons, William M. Jr.; Brodersen, Ditlev E.; McCutcheonn, John P.; May, Joanna L.C.; Carter, Andrew P.; Morgan-Warren, Robert J.; Wimberly, Brian T.; Ramakrishnan, Venki

    2001-01-01

    We describe the crystallization and structure determination of the 30 S ribosomal subunit from Thermus thermophilus. Previous reports of crystals that diffracted to 10 (angstrom) resolution were used as a starting point to improve the quality of the diffraction. Eventually, ideas such as the addition of substrates or factors to eliminate conformational heterogeneity proved less important than attention to detail in yielding crystals that diffracted beyond 3 (angstrom) resolution. Despite improvements in technology and methodology in the last decade, the structure determination of the 30 S subunit presented some very challenging technical problems because of the size of the asymmetric unit, crystal variability and sensitivity to radiation damage. Some steps that were useful for determination of the atomic structure were: the use of anomalous scattering from the LIII edges of osmium and lutetium to obtain the necessary phasing signal; the use of tunable, third-generation synchrotron sources to obtain data of reasonable quality at high resolution; collection of derivative data precisely about a mirror plane to preserve small anomalous differences between Bijvoet mates despite extensive radiation damage and multi-crystal scaling; the pre-screening of crystals to ensure quality, isomorphism and the efficient use of scarce third-generation synchrotron time; pre-incubation of crystals in cobalt hexaammine to ensure isomorphism with other derivatives; and finally, the placement of proteins whose structures had been previously solved in isolation, in conjunction with biochemical data on protein-RNA interactions, to map out the architecture of the 30 S subunit prior to the construction of a detailed atomic-resolution model.

  1. Role of disulphide bonds in a thermophilic serine protease aqualysin I from Thermus aquaticus YT-1.

    Science.gov (United States)

    Sakaguchi, Masayoshi; Takezawa, Makoto; Nakazawa, Rie; Nozawa, Kazutaka; Kusakawa, Taro; Nagasawa, Takeshi; Sugahara, Yasusato; Kawakita, Masao

    2008-05-01

    A thermophilic serine protease, Aqualysin I, from Thermus aquaticus YT-1 has two disulphide bonds, which are also found in a psychrophilic serine protease from Vibrio sp. PA-44 and a proteinase K-like enzyme from Serratia sp. at corresponding positions. To understand the significance of these disulphide bonds in aqualysin I, we prepared mutants C99S, C194S and C99S/C194S (WSS), in which Cys69-Cys99, Cys163-Cys194 and both of these disulphide bonds, respectively, were disrupted by replacing Cys residues with Ser residues. All mutants were expressed stably in Escherichia coli. The C99S mutant was 68% as active as the wild-type enzyme at 40 degrees C in terms of k(cat) value, while C194S and WSS were only 6 and 3%, respectively, as active, indicating that disulphide bond Cys163-Cys194 is critically important for maintaining proper catalytic site conformation. Mutants C194S and WSS were less thermostable than wild-type enzyme, with a half-life at 90 degrees C of 10 min as compared to 45 min of the latter and with transition temperatures on differential scanning calorimetry of 86.7 degrees C and 86.9 degrees C, respectively. Mutant C99S was almost as stable as the wild-type aqualysin I. These results indicate that the disulphide bond Cys163-Cys194 is more important for catalytic activity and conformational stability of aqualysin I than Cys67-Cys99.

  2. Roles of conserved arginines in ATP-binding domains of AAA+ chaperone ClpB from Thermus thermophilus.

    Science.gov (United States)

    Yamasaki, Takashi; Nakazaki, Yosuke; Yoshida, Masasuke; Watanabe, Yo-hei

    2011-07-01

    ClpB, a member of the expanded superfamily of ATPases associated with diverse cellular activities (AAA+), forms a ring-shaped hexamer and cooperates with the DnaK chaperone system to reactivate aggregated proteins in an ATP-dependent manner. The ClpB protomer consists of an N-terminal domain, an AAA+ module (AAA-1), a middle domain, and a second AAA+ module (AAA-2). Each AAA+ module contains highly conserved WalkerA and WalkerB motifs, and two arginines (AAA-1) or one arginine (AAA-2). Here, we investigated the roles of these arginines (Arg322, Arg323, and Arg747) of ClpB from Thermus thermophilus in the ATPase cycle and chaperone function by alanine substitution. These mutations did not affect nucleotide binding, but did inhibit the hydrolysis of the bound ATP and slow the threading of the denatured protein through the central pore of the T. thermophilus ClpB ring, which severely impaired the chaperone functions. Previously, it was demonstrated that ATP binding to the AAA-1 module induced motion of the middle domain and stabilized the ClpB hexamer. However, the arginine mutations of the AAA-1 module destabilized the ClpB hexamer, even though ATP-induced motion of the middle domain was not affected. These results indicated that the three arginines are crucial for ATP hydrolysis and chaperone activity, but not for ATP binding. In addition, the two arginines in AAA-1 and the ATP-induced motion of the middle domain independently contribute to the stabilization of the hexamer. © 2011 The Authors Journal compilation © 2011 FEBS.

  3. Properties of a novel thermostable glucose isomerase mined from Thermus oshimai and its application to preparation of high fructose corn syrup.

    Science.gov (United States)

    Jia, Dong-Xu; Zhou, Lin; Zheng, Yu-Guo

    2017-04-01

    Glucose isomerase (GI) is used in vitro to convert d-glucose to d-fructose, which is capable of commercial producing high fructose corn syrup (HFCS). To manufacture HFCS at elevated temperature and reduce the cost of enriching syrups, novel refractory GIs from Thermoanaerobacterium xylanolyticum (TxGI), Thermus oshimai (ToGI), Geobacillus thermocatenulatus (GtGI) and Thermoanaerobacter siderophilus (TsGI) were screened via genome mining approach. The enzymatic characteristics research showed that ToGI had higher catalytic efficiency and superior thermostability toward d-glucose among the screened GIs. Its optimum temperature reached 95°C and could retain more than 80% of initial activity in the presence of 20mM Mn 2+ at 85°C for 48h. The K m and k cat /K m values for ToGI were 81.46mM and 21.77min -1 mM -1 , respectively. Furthermore, the maximum conversion yield of 400g/L d-glucose to d-fructose at 85°C was 52.16%. Considering its excellent high thermostability and ameliorable application performance, ToGI might be promising for realization of future industrial production of HFCS at elevated temperature. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Modelling the structure and kinematics of the Firework nebula: The nature of the GK Persei nova shell and its jet-like feature

    Science.gov (United States)

    Harvey, E.; Redman, M. P.; Boumis, P.; Akras, S.

    2016-10-01

    Aims: The shaping mechanisms of old nova remnants are probes for several important and unexplained processes, such as dust formation and the structure of evolved star nebulae. To gain a more complete understanding of the dynamics of the GK Per (1901) remnant, an examination of symmetry of the nova shell is explored, followed by a kinematical analysis of the previously detected jet-like feature in the context of the surrounding fossil planetary nebula. Methods: Faint-object high-resolution echelle spectroscopic observations and imaging were undertaken covering the knots which comprise the nova shell and the surrounding nebulosity. New imaging from the Aristarchos telescope in Greece and long-slit spectra from the Manchester Echelle Spectrometer instrument at the San Pedro Mártir observatory in Mexico were obtained, supplemented with archival observations from several other optical telescopes. Position-velocity arrays are produced of the shell, and also individual knots, and are then used for morpho-kinematic modelling with the shape code. The overall structure of the old knotty nova shell of GK Per and the planetary nebula in which it is embedded is then analysed. Results: Evidence is found for the interaction of knots with each other and with a wind component, most likely the periodic fast wind emanating from the central binary system. We find that a cylindrical shell with a lower velocity polar structure gives the best model fit to the spectroscopy and imaging. We show in this work that the previously seen jet-like feature is of low velocity. Conclusions: The individual knots have irregular tail shapes; we propose here that they emanate from episodic winds from ongoing dwarf nova outbursts by the central system. The nova shell is cylindrical, not spherical, and the symmetry axis relates to the inclination of the central binary system. Furthermore, the cylinder axis is aligned with the long axis of the bipolar planetary nebula in which it is embedded. Thus, the

  5. Glycerol rhizotomy versus gamma knife radiosurgery for the treatment of trigeminal neuralgia: An analysis of patients treated at one institution

    International Nuclear Information System (INIS)

    Henson, Clarissa Febles; Goldman, H. Warren; Rosenwasser, Robert H.; Downes, M. Beverly; Bednarz, Greg; Pequignot, Edward C.; Werner-Wasik, Maria; Curran, Walter J.; Andrews, David W.

    2005-01-01

    .05. Results: There were 63 evaluable GK patients and 36 evaluable GR patients. The median follow-up time was 34 and 29 months for the GR and GK groups, respectively. The median age was 69 and 70 years and the median number of years with TN pain was 6 and 7 years in the GR and GK groups, respectively. Thirty-one GR (86%) and 58 GK (92%) patients achieved a successful treatment outcome (p = 0.49). The median time to pain relief was ≤24 h in the GR group and 3 weeks in the GK group (p < 0.001, ordinal logistic regression). Nineteen GR and 26 GK patients experienced pain recurrence or pain never relieved (p = 0.30). The median time to pain recurrence was 5 and 8 months in the GR and GK groups, respectively (p = 0.22). At last follow-up, however, a statistically significant greater number of GR vs. GK patients had failed treatment. Twelve or 33% of GR patients, whereas 8 or 13% of GK patients, had BNI scores of 4 or 5 (p = 0.019, Fisher's exact test). When both initial and late treatment failures were combined, the overall rate of treatment failures was 39% in the GR group and 24% in the GK group (p = 0.023, log-rank). More GR patients developed facial numbness and facial numbness morbidity than GK patients: 19 GR (54%) and 17 GK patients (30%) developed new facial numbness and 12 GR and 7 GK patients reported either somewhat or very bothersome facial numbness (p = 0.018). On multivariate analysis, only treatment with GK and severity of pain before treatment significantly predicted treatment response. GK patients were more likely to have a lower pain score at last follow-up than were GR patients (p = 0.005, OR = 4.3), and patients with pretreatment pain scores of 5 were more likely to have lower posttreatment pain scores than patients with pretreatment pain scores of 4 and lower (p = 0.003, OR = 4.0). Conclusion: Despite greater facial numbness morbidity and a higher failure rate, GR provided more immediate pain relief than GK. GR therefore should be considered in patients with

  6. RNA and DNA Targeting by a Reconstituted Thermus thermophilus Type III-A CRISPR-Cas System.

    Directory of Open Access Journals (Sweden)

    Tina Y Liu

    Full Text Available CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR-associated systems are RNA-guided adaptive immunity pathways used by bacteria and archaea to defend against phages and plasmids. Type III-A systems use a multisubunit interference complex called Csm, containing Cas proteins and a CRISPR RNA (crRNA to target cognate nucleic acids. The Csm complex is intriguing in that it mediates RNA-guided targeting of both RNA and transcriptionally active DNA, but the mechanism is not well understood. Here, we overexpressed the five components of the Thermus thermophilus (T. thermophilus Type III-A Csm complex (TthCsm with a defined crRNA sequence, and purified intact TthCsm complexes from E. coli cells. The complexes were thermophilic, targeting complementary ssRNA more efficiently at 65°C than at 37°C. Sequence-independent, endonucleolytic cleavage of single-stranded DNA (ssDNA by TthCsm was triggered by recognition of a complementary ssRNA, and required a lack of complementarity between the first 8 nucleotides (5' tag of the crRNA and the 3' flanking region of the ssRNA. Mutation of the histidine-aspartate (HD nuclease domain of the TthCsm subunit, Cas10/Csm1, abolished DNA cleavage. Activation of DNA cleavage was dependent on RNA binding but not cleavage. This leads to a model in which binding of an ssRNA target to the Csm complex would stimulate cleavage of exposed ssDNA in the cell, such as could occur when the RNA polymerase unwinds double-stranded DNA (dsDNA during transcription. Our findings establish an amenable, thermostable system for more in-depth investigation of the targeting mechanism using structural biology methods, such as cryo-electron microscopy and x-ray crystallography.

  7. RNA and DNA Targeting by a Reconstituted Thermus thermophilus Type III-A CRISPR-Cas System.

    Science.gov (United States)

    Liu, Tina Y; Iavarone, Anthony T; Doudna, Jennifer A

    2017-01-01

    CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR-associated) systems are RNA-guided adaptive immunity pathways used by bacteria and archaea to defend against phages and plasmids. Type III-A systems use a multisubunit interference complex called Csm, containing Cas proteins and a CRISPR RNA (crRNA) to target cognate nucleic acids. The Csm complex is intriguing in that it mediates RNA-guided targeting of both RNA and transcriptionally active DNA, but the mechanism is not well understood. Here, we overexpressed the five components of the Thermus thermophilus (T. thermophilus) Type III-A Csm complex (TthCsm) with a defined crRNA sequence, and purified intact TthCsm complexes from E. coli cells. The complexes were thermophilic, targeting complementary ssRNA more efficiently at 65°C than at 37°C. Sequence-independent, endonucleolytic cleavage of single-stranded DNA (ssDNA) by TthCsm was triggered by recognition of a complementary ssRNA, and required a lack of complementarity between the first 8 nucleotides (5' tag) of the crRNA and the 3' flanking region of the ssRNA. Mutation of the histidine-aspartate (HD) nuclease domain of the TthCsm subunit, Cas10/Csm1, abolished DNA cleavage. Activation of DNA cleavage was dependent on RNA binding but not cleavage. This leads to a model in which binding of an ssRNA target to the Csm complex would stimulate cleavage of exposed ssDNA in the cell, such as could occur when the RNA polymerase unwinds double-stranded DNA (dsDNA) during transcription. Our findings establish an amenable, thermostable system for more in-depth investigation of the targeting mechanism using structural biology methods, such as cryo-electron microscopy and x-ray crystallography.

  8. ORF Alignment: NC_005835 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available [Thermus thermophilus HB8] pdb|1FP9|A Chain A, Structure ... Of Amylomaltase From Thermus Thermophilus Hb8 In Space... ... Amylomaltase From Thermus Thermophilus Hb8 In Space ... Group P21212 ... Length = 500 ... Query

  9. ORF Alignment: NC_006461 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available [Thermus thermophilus HB8] pdb|1FP9|A Chain A, Structure ... Of Amylomaltase From Thermus Thermophilus Hb8 In Space... ... Amylomaltase From Thermus Thermophilus Hb8 In Space ... Group P21212 ... Length = 500 ... Query

  10. Insight into the transition between the open and closed conformations of Thermus thermophilus carboxypeptidase

    International Nuclear Information System (INIS)

    Okai, Masahiko; Yamamura, Akihiro; Hayakawa, Kou; Tsutsui, Shiho; Miyazono, Ken-ichi; Lee, Woo-Cheol; Nagata, Koji; Inoue, Yumiko; Tanokura, Masaru

    2017-01-01

    Carboxypeptidase cleaves the C-terminal amino acid residue from proteins and peptides. Here, we report the functional and structural characterizations of carboxypeptidase belonging to the M32 family from the thermophilic bacterium Thermus thermophilus HB8 (TthCP). TthCP exhibits a relatively broad specificity for both hydrophilic (neutral and basic) and hydrophobic (aliphatic and aromatic) residues at the C-terminus and shows optimal activity in the temperature range of 75–80 °C and in the pH range of 6.8–7.2. Enzyme activity was significantly enhanced by cobalt or cadmium and was moderately inhibited by Tris at 25 °C. We also determined the crystal structure of TthCP at 2.6 Å resolution. Two dimer types of TthCP are present in the crystal. One type consists of two subunits in different states, open and closed, with a C α RMSD value of 2.2 Å; the other type consists of two subunits in the same open state. This structure enables us to compare the open and closed states of an M32 carboxypeptidase. The TthCP subunit can be divided into two domains, L and S, which are separated by a substrate-binding groove. The L and S domains in the open state are almost identical to those in the closed state, with C α RMSD values of 0.84 and 0.53 Å, respectively, suggesting that the transition between the open and closed states proceeds with a large hinge-bending motion. The superimposition between the closed states of TthCP and BsuCP, another M32 family member, revealed that most putative substrate-binding residues in the grooves are oriented in the same direction. - Highlights: • The enzyme activity of TthCP was inhibited moderately by Tris molecule. • We solved the crystal structure of TthCP at 2.6 Å resolution. • The crystal structure of TthCP revealed both the open and closed conformations.

  11. Fluid balance and dehydration in futsal players: goalkeepers vs. field players. (Reposición hídrica y deshidratación en jugadores de fútbol sala: porteros vs. jugadores de campo.

    Directory of Open Access Journals (Sweden)

    Juan José García-Pellicer

    2011-01-01

    Full Text Available AbstractDehydration higher than 2% can adversely affect performance. The purpose of this study was to determine and to compare fluid balance in elite futsal players in relation to playing position. We tested three goalkeepers (GK and nine field players (FP during 3 official matches (22-24 ºC and 39.33-42% RH. Body mass changes, fluid intake (ad libitum and urine loss were calculated. To analyze this data the statistics used are descriptive and no parametric. Sweat lost in FP (2450±774.63 ml. was higher than in GK (2195±557.64 ml.. Fluid intake in FP (1650.42±774.74 ml. was higher than in GK (1205±333.31 ml.. Dehydration in GK (1.27±0.60% was higher than FP (1.00±1.15%. There were no significant difference in body weight loss and sweat lost replaced (p>0.05 in relation to playing position. Even GK and FP dehydration status are not associated with a reduction in performance. In these players, independently of position, hydration status can be seen to maintain itself through regular substitutions and also the correct fluid intake.ResumenUn nivel de deshidratación superior al 2% puede afectar negativamente al rendimiento. El objetivo de este estudio ha sido determinar y comparar la reposición hídrica en jugadores profesionales de fútbol sala según su posición. 3 porteros (GK y 9 jugadores de campo (FP fueron estudiados durante 3 partidos oficiales (22-24 ºC y 39,33-42% HR. Se registraron los cambios en el peso corporal, líquido ingerido (ad libitum y orina excretada. Para el análisis de datos se aplicó estadística descriptiva y no paramétrica. El sudor perdido en FP (2450±774,63 ml. fue mayor que en GK (2195±557.64 ml.. El líquido ingerido en FP (1650,42±774,74 ml. fue mayor que en GK (1205±333,31 ml.. La deshidratación en GK (1,27±0,60% fue mayor que en FP (1,00±1,15%. No existen diferencias en los valores de deshidratación y líquido perdido (p>0,05 en función del puesto específico. El nivel de deshidrataci

  12. Patronage Politics and Clientelism in Housing Welfare: The Case of Gawad Kalinga (GK Villages in Parañaque City, The Philippines

    Directory of Open Access Journals (Sweden)

    Prince Aian G. Villanueva

    2015-08-01

    Full Text Available Patronage politics has become a defining charac-teristic of the Philippine political landscape. Clientelism, a form of patronage, is often studied as disadvantageous in governance and democratic consolidation. While the patron-client framework remains the most influential among schools of thought that explain Philippine politics, transforma-tions in a clientelist exchange are evident given changes in political, cultural and economic settings. Using this frame of new clientelism, we look at and revisit the structure of patronage in the provision of housing welfare in the Philippines, focusing now on the roles of three important actors— the nongov-ernmental organization, the state through the local government and the recipients or beneficiaries— in an urban setting, thus deviating from the traditional conception of patronage and clientelist politics. This relationship of clientelist exchange is presented in our Accomplice-Principal-Accessory (APA model ofclientelism, with the local government unit of Parañaque City as accomplice, the Gawad Kalinga (GK as the principal agent, and the recipients or beneficiaries as accessories in the provision of hous-ing welfare.

  13. Acarviosine-simmondsin, a novel compound obtained from acarviosine-glucose and simmondsin by Thermus maltogenic amylase and its in vivo effect on food intake and hyperglycemia.

    Science.gov (United States)

    Baek, Jin-Sook; Kim, Hye-Young; Abbott, Thomas P; Moon, Tae-Wha; Lee, Soo-Bok; Park, Cheon-Seok; Park, Kwan-Hwa

    2003-03-01

    Simmondsin was modified with acarviosine-glucose using the transglycosylation activity of Thermus maltogenic amylase to synthesize a novel compound with both antiobesity and hypoglycemic activity. The LC/MS and 13C NMR analyses confirmed that the structure of the major transglycosylation product was acarviosine-simmondsin (Acv-simmondsin), in which acarviosine was attached to the glucose moiety of simmondsin by an alpha-(1,6)-glycosidic linkage. It was found that Acv-simmondsin was a potent competitive inhibitor of alpha-glucosidase with the Ki value of 0.69 microM and a mixed type inhibitor of alpha-amylase with the Ki and KI of 20.78 microM and 26.31 microM, respectively. The administration of Acv-simmondsin (0.1 g/100 g diet/day) to mice for 5 days significantly reduced food intake by 35%, compared to 25% with simmondsin in control obese mice. Acv-simmondsin (50 mg/kg BW) suppressed the postprandial blood glucose response to sucrose (1 g/kg BW) by 74%, compared to 71% with acarbose, in normal rats.

  14. Cre/lox-based multiple markerless gene disruption in the genome of the extreme thermophile Thermus thermophilus.

    Science.gov (United States)

    Togawa, Yoichiro; Nunoshiba, Tatsuo; Hiratsu, Keiichiro

    2018-02-01

    Markerless gene-disruption technology is particularly useful for effective genetic analyses of Thermus thermophilus (T. thermophilus), which have a limited number of selectable markers. In an attempt to develop a novel system for the markerless disruption of genes in T. thermophilus, we applied a Cre/lox system to construct a triple gene disruptant. To achieve this, we constructed two genetic tools, a loxP-htk-loxP cassette and cre-expressing plasmid, pSH-Cre, for gene disruption and removal of the selectable marker by Cre-mediated recombination. We found that the Cre/lox system was compatible with the proliferation of the T. thermophilus HB27 strain at the lowest growth temperature (50 °C), and thus succeeded in establishing a triple gene disruptant, the (∆TTC1454::loxP, ∆TTC1535KpnI::loxP, ∆TTC1576::loxP) strain, without leaving behind a selectable marker. During the process of the sequential disruption of multiple genes, we observed the undesired deletion and inversion of the chromosomal region between multiple loxP sites that were induced by Cre-mediated recombination. Therefore, we examined the effects of a lox66-htk-lox71 cassette by exploiting the mutant lox sites, lox66 and lox71, instead of native loxP sites. We successfully constructed a (∆TTC1535::lox72, ∆TTC1537::lox72) double gene disruptant without inducing the undesired deletion of the 0.7-kbp region between the two directly oriented lox72 sites created by the Cre-mediated recombination of the lox66-htk-lox71 cassette. This is the first demonstration of a Cre/lox system being applicable to extreme thermophiles in a genetic manipulation. Our results indicate that this system is a powerful tool for multiple markerless gene disruption in T. thermophilus.

  15. Purification and characterization of a novel recombinant highly enantioselective short-chain NAD(H)-dependent alcohol dehydrogenase from Thermus thermophilus.

    Science.gov (United States)

    Pennacchio, Angela; Pucci, Biagio; Secundo, Francesco; La Cara, Francesco; Rossi, Mosè; Raia, Carlo A

    2008-07-01

    The gene encoding a novel alcohol dehydrogenase (ADH) that belongs to the short-chain dehydrogenase/reductase (SDR) superfamily was identified in the extremely thermophilic, halotolerant gram-negative eubacterium Thermus thermophilus HB27. The T. thermophilus ADH gene (adh(Tt)) was heterologously overexpressed in Escherichia coli, and the protein (ADH(Tt)) was purified to homogeneity and characterized. ADH(Tt) is a tetrameric enzyme consisting of identical 26,961-Da subunits composed of 256 amino acids. The enzyme has remarkable thermophilicity and thermal stability, displaying activity at temperatures up to approximately 73 degrees C and a 30-min half-inactivation temperature of approximately 90 degrees C, as well as good tolerance to common organic solvents. ADH(Tt) has a strict requirement for NAD(H) as the coenzyme, a preference for reduction of aromatic ketones and alpha-keto esters, and poor activity on aromatic alcohols and aldehydes. This thermophilic enzyme catalyzes the following reactions with Prelog specificity: the reduction of acetophenone, 2,2,2-trifluoroacetophenone, alpha-tetralone, and alpha-methyl and alpha-ethyl benzoylformates to (S)-(-)-1-phenylethanol (>99% enantiomeric excess [ee]), (R)-alpha-(trifluoromethyl)benzyl alcohol (93% ee), (S)-alpha-tetralol (>99% ee), methyl (R)-(-)-mandelate (92% ee), and ethyl (R)-(-)-mandelate (95% ee), respectively, by way of an efficient in situ NADH-recycling system involving 2-propanol and a second thermophilic ADH. This study further supports the critical role of the D37 residue in discriminating NAD(H) from NADP(H) in members of the SDR superfamily.

  16. Crystallization and preliminary X-ray diffraction analysis of recombinant phosphoribosylpyrophosphate synthetase from the Thermophilic thermus thermophilus strain HB27

    Energy Technology Data Exchange (ETDEWEB)

    Abramchik, Yu. A. [Russian Academy of Sciences, Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry (Russian Federation); Timofeev, V. I., E-mail: tostars@mail.ru [Russian Academy of Sciences, Shubnikov Institute of Crystallography of Federal Scientific Research Centre “Crystallography and Photonics” (Russian Federation); Muravieva, T. I.; Sinitsyna, E. V.; Esipov, R. S., E-mail: esipov@mx.ibch.ru [Russian Academy of Sciences, Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry (Russian Federation); Kuranova, I. P., E-mail: inna@ns.crys.ras.ru [Russian Academy of Sciences, Shubnikov Institute of Crystallography of Federal Scientific Research Centre “Crystallography and Photonics” (Russian Federation)

    2017-01-15

    Phosphoribosylpyrophosphate synthetases (PRPP synthetases) are among the key enzymes essential for vital functions of organisms and are involved in the biosynthesis of purine and pyrimidine nucleotides, coenzymes, and the amino acids histidine and tryptophan. These enzymes are used in biotechnology for the combined chemoenzymatic synthesis of natural nucleotide analogs. Recombinant phosphoribosylpyrophosphate synthetase I from the thermophilic strain HB27 of the bacterium Thermus thermophilus (T. th HB27) has high thermal stability and shows maximum activity at 75°Ð¡, due to which this enzyme holds promise for biotechnological applications. In order to grow crystals and study them by X-ray crystallography, an enzyme sample, which was produced using a highly efficient producer strain, was purified by affinity and gel-filtration chromatography. The screening of crystallization conditions was performed by the vapor-diffusion technique. The crystals of the enzyme suitable for X-ray diffraction were grown by the counter-diffusion method through a gel layer. These crystals were used to collect the X-ray diffraction data set at the SPring-8 synchrotron radiation facility (Japan) to 3-Å resolution. The crystals belong to sp. gr. P2{sub 1} and have the following unitcell parameters: a = 107.7 Å, b = 112.6 Å, c = 110.2 Å, α = γ = 90°, β = 116.6°. The X-ray diffraction data set is suitable for determining the three-dimensional structure of the enzyme at 3.0-Å resolution.

  17. Crystal structure studies of NADP{sup +} dependent isocitrate dehydrogenase from Thermus thermophilus exhibiting a novel terminal domain

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, S.M. [Department of Studies in Physics, University of Mysore, Mysore 570 006 (India); Pampa, K.J. [Department of Studies in Microbiology, University of Mysore, Mysore 570 006 (India); Manjula, M. [Department of Studies in Physics, University of Mysore, Mysore 570 006 (India); Abdoh, M.M.M. [Department of Physics, Faculty of Science, An-Najah National University, Nablus, West Bank, Palestine (Country Unknown); Kunishima, Naoki [Advanced Protein Crystallography Research Group, RIKEN SPring-8 Center, Harima Institute, Hyogo 679-5148 (Japan); Lokanath, N.K., E-mail: lokanath@physics.uni-mysore.ac.in [Department of Studies in Physics, University of Mysore, Mysore 570 006 (India)

    2014-06-20

    Highlights: • We determined the structure of isocitrate dehydrogenase with citrate and cofactor. • The structure reveals a unique novel terminal domain involved in dimerization. • Clasp domain shows significant difference, and catalytic residues are conserved. • Oligomerization of the enzyme is quantized with subunit-subunit interactions. • Novel domain of this enzyme is classified as subfamily of the type IV. - Abstract: NADP{sup +} dependent isocitrate dehydrogenase (IDH) is an enzyme catalyzing oxidative decarboxylation of isocitrate into oxalosuccinate (intermediate) and finally the product α-ketoglutarate. The crystal structure of Thermus thermophilus isocitrate dehydrogenase (TtIDH) ternary complex with citrate and cofactor NADP{sup +} was determined using X-ray diffraction method to a resolution of 1.80 Å. The overall fold of this protein was resolved into large domain, small domain and a clasp domain. The monomeric structure reveals a novel terminal domain involved in dimerization, very unique and novel domain when compared to other IDH’s. And, small domain and clasp domain showing significant differences when compared to other IDH’s of the same sub-family. The structure of TtIDH reveals the absence of helix at the clasp domain, which is mainly involved in oligomerization in other IDH’s. Also, helices/beta sheets are absent in the small domain, when compared to other IDH’s of the same sub family. The overall TtIDH structure exhibits closed conformation with catalytic triad residues, Tyr144-Asp248-Lys191 are conserved. Oligomerization of the protein is quantized using interface area and subunit–subunit interactions between protomers. Overall, the TtIDH structure with novel terminal domain may be categorized as a first structure of subfamily of type IV.

  18. Electrodes for 24 hours pH monitoring--a comparative study.

    OpenAIRE

    McLauchlan, G; Rawlings, J M; Lucas, M L; McCloy, R F; Crean, G P; McColl, K E

    1987-01-01

    Three pH electrodes in clinical use were examined--(1) antimony electrode with remote reference electrode (Synectics 0011), (2) glass electrode with remote reference electrode (Microelectrodes Inc. MI 506) and (3) combined glass electrode with integral reference electrode (Radiometer GK2801C). In vitro studies showed that both glass electrodes were similar and superior to the antimony electrode with respect to response time, drift, and sensitivity. The effect of the siting of the reference el...

  19. NCBI nr-aa BLAST: CBRC-TBEL-01-0397 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TBEL-01-0397 ref|YP_005572.1| competence factor comEC [Thermus thermophilus HB...27] gb|AAG34707.1|AF319938_2 competence factor ComEC [Thermus thermophilus] gb|AAS81945.1| competence factor comEC [Thermus thermophilus HB27] YP_005572.1 0.10 31% ...

  20. The role of the PHP domain associated with DNA polymerase X from Thermus thermophilus HB8 in base excision repair.

    Science.gov (United States)

    Nakane, Shuhei; Nakagawa, Noriko; Kuramitsu, Seiki; Masui, Ryoji

    2012-11-01

    Base excision repair (BER) is one of the most commonly used DNA repair pathways involved in genome stability. X-family DNA polymerases (PolXs) play critical roles in BER, especially in filling single-nucleotide gaps. In addition to a polymerase core domain, bacterial PolXs have a polymerase and histidinol phosphatase (PHP) domain with phosphoesterase activity which is also required for BER. However, the role of the PHP domain of PolX in bacterial BER remains unresolved. We found that the PHP domain of Thermus thermophilus HB8 PolX (ttPolX) functions as two types of phosphoesterase in BER, including a 3'-phosphatase and an apurinic/apyrimidinic (AP) endonuclease. Experiments using T. thermophilus HB8 cell lysates revealed that the majority of the 3'-phosphatase and AP endonuclease activities are attributable to the another phosphoesterase in T. thermophilus HB8, endonuclease IV (ttEndoIV). However, ttPolX possesses significant 3'-phosphatase activity in ΔttendoIV cell lysate, indicating possible complementation. Our experiments also reveal that there are only two enzymes that display the 3'-phosphatase activity in the T. thermophilus HB8 cell, ttPolX and ttEndoIV. Furthermore, phenotypic analysis of ΔttpolX, ΔttendoIV, and ΔttpolX/ΔttendoIV using hydrogen peroxide and sodium nitrite supports the hypothesis that ttPolX functions as a backup for ttEndoIV in BER. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Crystallization and preliminary X-ray diffraction study of recombinant adenine phosphoribosyltransferase from the thermophilic bacterium Thermus thermophilus strain HB27

    Science.gov (United States)

    Sinitsyna, E. V.; Timofeev, V. I.; Tuzova, E. S.; Kostromina, M. A.; Murav'eva, T. I.; Esipov, R. S.; Kuranova, I. P.

    2017-07-01

    Adenine phosphoribosyltransferase (APRT) belongs to the type I phosphoribosyltransferase family and catalyzes the formation of adenosine monophosphate via transfer of the 5-phosphoribosyl group from phosphoribosyl pyrophosphate to the nitrogen atom N9 of the adenine base. Proteins of this family are involved in a salvage pathway of nucleotide synthesis, thus providing purine base utilization and maintaining the optimal level of purine bases in the body. Adenine phosphoribosyltransferase from the extremely thermophilic Thermus thermophilus strain HB27 was produced using a highly efficient E. coli producer strain and was then purified by affinity and gel-filtration chromatography. This enzyme was successfully employed as a catalyst for the cascade biosynthesis of biologically important nucleotides. The screening of crystallization conditions for recombinant APRT from T. thermophilus HB27 was performed in order to determine the enzyme structure by X-ray diffraction. The crystallization conditions, which were found by the vapor-diffusion technique, were then optimized to apply the counter-diffusion technique. The crystals of the enzyme were grown by the capillary counter-diffusion method. The crystals belong to sp. gr. P1211 and have the following unitcell parameters: a = 69.86 Å, b = 82.16 Å, c = 91.39 Å, α = γ = 90°, β = 102.58°. The X-ray diffraction data set suitable for the determination of the APRT structure at 2.6 Å resolution was collected from the crystals at the SPring-8 synchrotron facility (Japan).

  2. Identification of novel esterase-active enzymes from hot environments by use of the host bacterium Thermus thermophilus

    Directory of Open Access Journals (Sweden)

    Benedikt eLeis

    2015-04-01

    Full Text Available Functional metagenomic screening strategies, which are independent of known sequence information, can lead to the identification of truly novel genes and enzymes. Since E. coli has been used exhaustively for this purpose as a host, it is important to establish alternative expression hosts and to use them for functional metagenomic screening for new enzymes. In this study we show that Thermus thermophilus HB27 is an excellent screening host and can be used as an alternative provider of truly novel biocatalysts. In a previous study we constructed the mutant strain BL03 that was no longer able to grow on defined minimal medium supplemented with tributyrin as the sole carbon source and could be used as a host to screen for metagenomic DNA fragments that could complement growth on tributyrin. Several thousand single fosmid clones from thermophilic metagenomic libraries from heated compost and hot spring water samples were subjected to a comparative screening for esterase activity in both T. thermophilus strain BL03 and E. coli EPI300. We scored a greater number of active clones in the thermophilic bacterium than in the mesophilic E. coli. From all clones functionally screened in E. coli, only two thermostable α/β-fold hydrolase enzymes with high amino acid sequence similarity to already characterized enzymes were identifiable. In contrast, five further fosmids were found that conferred lipolytic activities in T. thermophilus. Four open reading frames (ORFs were found which did not share significant similarity to known esterase enzymes. Two of the genes were expressed in both hosts and the novel thermophilic esterases, which based on their primary structures could not be assigned to known esterase or lipase families, were purified and preliminarily characterized. Our work underscores the benefit of using additional screening hosts other than E. coli for the identification of novel biocatalysts with industrial relevance.

  3. A Flexible Domain-Domain Hinge Promotes an Induced-fit Dominant Mechanism for the Loading of Guide-DNA into Argonaute Protein in Thermus thermophilus

    KAUST Repository

    Zhu, Lizhe

    2016-02-24

    Argonaute proteins (Ago) are core components of the RNA Induced Silencing Complex (RISC) that load and utilize small guide nucleic acids to silence mRNAs or cleave foreign DNAs. Despite the essential role of Ago in gene regulation and defense against virus, the molecular mechanism of guide-strand loading into Ago remains unclear. We explore such a mechanism in the bacterium Thermus thermophilus Ago (TtAgo), via a computational approach combining molecular dynamics, bias-exchange metadynamics, and protein-DNA docking. We show that apo TtAgo adopts multiple closed states that are unable to accommodate guide-DNA. Conformations able to accommodate the guide are beyond the reach of thermal fluctuations from the closed states. These results suggest an induced-fit dominant mechanism for guide-strand loading in TtAgo, drastically different from the two-step mechanism for human Ago 2 (hAgo2) identified in our previous study. Such a difference between TtAgo and hAgo2 is found to mainly originate from the distinct rigidity of their L1-PAZ hinge. Further comparison among known Ago structures from various species indicates that the L1-PAZ hinge may be flexible in general for prokaryotic Agos but rigid for eukaryotic Agos. © 2016 American Chemical Society.

  4. A Flexible Domain-Domain Hinge Promotes an Induced-fit Dominant Mechanism for the Loading of Guide-DNA into Argonaute Protein in Thermus thermophilus.

    Science.gov (United States)

    Zhu, Lizhe; Jiang, Hanlun; Sheong, Fu Kit; Cui, Xuefeng; Gao, Xin; Wang, Yanli; Huang, Xuhui

    2016-03-17

    Argonaute proteins (Ago) are core components of the RNA Induced Silencing Complex (RISC) that load and utilize small guide nucleic acids to silence mRNAs or cleave foreign DNAs. Despite the essential role of Ago in gene regulation and defense against virus, the molecular mechanism of guide-strand loading into Ago remains unclear. We explore such a mechanism in the bacterium Thermus thermophilus Ago (TtAgo), via a computational approach combining molecular dynamics, bias-exchange metadynamics, and protein-DNA docking. We show that apo TtAgo adopts multiple closed states that are unable to accommodate guide-DNA. Conformations able to accommodate the guide are beyond the reach of thermal fluctuations from the closed states. These results suggest an induced-fit dominant mechanism for guide-strand loading in TtAgo, drastically different from the two-step mechanism for human Ago 2 (hAgo2) identified in our previous study. Such a difference between TtAgo and hAgo2 is found to mainly originate from the distinct rigidity of their L1-PAZ hinge. Further comparison among known Ago structures from various species indicates that the L1-PAZ hinge may be flexible in general for prokaryotic Ago's but rigid for eukaryotic Ago's.

  5. Thermo-reversible inhibition makes aqualysin 1 from Thermus aquaticus a potent tool for studying the contribution of the wheat gluten network to the crumb texture of fresh bread.

    Science.gov (United States)

    Verbauwhede, Annelien E; Lambrecht, Marlies A; Fierens, Ellen; Hermans, Senne; Shegay, Oksana; Brijs, Kristof; Delcour, Jan A

    2018-10-30

    The thermo-active serine peptidase aqualysin 1 (Aq1) of Thermus aquaticus was applied in bread making to study the relative contribution of thermoset gluten to bread crumb texture. Aq1 is active between 30 °C and 90 °C with an optimum activity temperature of around 65 °C. It is inhibited by wheat endogenous serine peptidase inhibitors during dough mixing and fermentation and starts hydrolyzing gluten proteins during baking above 80 °C when the enzyme is no longer inhibited and most of the starch is gelatinized and contributes to structure formation. Aq1 activity reduced the molecular weight of gluten proteins and significantly increased their extractability in sodium dodecyl sulfate containing medium. While it had no impact on the specific bread volume and only limited impact on hardness, cohesiveness, springiness, resilience and chewiness, it impacted bread crumb coherence. We conclude that starch has a greater impact on crumb texture than thermoset gluten. Copyright © 2018 Elsevier Ltd. All rights reserved.

  6. ORF Alignment: NC_002935 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ... [Haloarcula marismortui ATCC 43049] ... Length = 295 ... Query: 24 ... KPFIAIWEVTRACGLVCQHCRADAQH...EPHPEQLTTEQGKQLLSDLASYERPKPLVVLTGG 83 ... +P++ IWEVT+AC L C+HCRADAQ ... HP++...L+T +GK LL D AS ... LVVL+GG Sbjct: 3 ... QPLVLIWEVTQACELACKHCRADAQPRRHPDELSTAEGKALL-DQASEFGDGQLVVLSGG 61 ... Que

  7. SU-D-BRB-04: Plan Quality Comparison of Intracranial Stereotactic Radiosurgery (SRS) for Gamma Knife and VMAT Treatments

    Energy Technology Data Exchange (ETDEWEB)

    Keeling, V; Algan, O; Ahmad, S; Hossain, S [University of Oklahoma Health Sciences Center, Oklahoma City, OK (United States)

    2015-06-15

    Purpose: To compare treatment plan quality of intracranial stereotactic radiosurgery (SRS) for VMAT (RapidArc) and Gamma Knife (GK) systems. Methods: Ten patients with 24 tumors (seven with 1–2 and three with 4–6 lesions), previously treated with GK 4C (prescription doses ranging from 14–23 Gy) were re-planned for RapidArc. Identical contour sets were kept on MRI images for both plans with tissues assigned a CT number of zero. RapidArc plans were performed using 6 MV flattening-filter-free (FFF) beams with dose rate of 1400 MU/minute using two to eight arcs with the following combinations: 2 full coplanar arcs and the rest non-coplanar half arcs. Beam selection was based on target depth. Areas that penetrated more than 10 cm of tissue were avoided by creating smaller arcs or using avoidance sectors in optimization. Plans were optimized with jaw tracking and a high weighting to the normal-brain-tissue and Normal-Tissue-Objective without compromising PTV coverage. Plans were calculated on a 1 mm grid size using AAA algorithm and then normalized so that 99% of each target volume received the prescription dose. Plan quality was assessed by target coverage using Paddick Conformity Index (PCI), sparing of normal-brain-tissue through analysis of V4, V8, and V12 Gy, and integral dose. Results: In all cases critical structure dose criteria were met. RapidArc had a higher PCI than GK plans for 23 out of 24 lesions. The average PCI was 0.76±0.21 for RapidArc and 0.46±0.20 for GK plans (p≤0.001), respectively. Integral dose and normal-brain-tissue doses for all criteria were lower for RapidArc in nearly all patients. The average ratio of GK to RapidArc plans was 1.28±0.27 (p=0.018), 1.31±0.25 (p=0.017), 1.81±0.43 (p=0.005), and 1.50±0.61 (p=0.006) for V4, V8, and V12 Gy, and integral dose, respectively. Conclusion: VMAT was capable of producing higher quality treatment plans than GK when using optimal beam geometries and proper optimization techniques.

  8. Measurement of key resonances for the 24Al(p ,γ )25Si reaction rate using in-beam γ -ray spectroscopy

    Science.gov (United States)

    Longfellow, B.; Gade, A.; Brown, B. A.; Richter, W. A.; Bazin, D.; Bender, P. C.; Bowry, M.; Elman, B.; Lunderberg, E.; Weisshaar, D.; Williams, S. J.

    2018-05-01

    Energy levels and branching ratios for the rp-process nucleus 25Si were determined from the reactions 9Be(26Si,25Si)X and 9Be(25Al,25Si)X using in-beam γ -ray spectroscopy with both high-efficiency and high-resolution detector arrays. Proton-unbound states at 3695(14) and 3802(11) keV were identified and assigned tentative spins and parities based on comparison to theory and the mirror nucleus. The 24Al(p ,γ )25Si reaction rate was calculated using the experimental states and states from charge-dependent USDA and USDB shell-model calculations with downward shifts of the 1 s1 /2 proton orbital to account for the observed Thomas-Ehrman shift, leading to a factor of 10-100 increase in rate for the temperature region of 0.22 GK as compared to a previous calculation. These shifts may be applicable to neighboring nuclei, impacting the proton capture rates in this region of the chart.

  9. Repeat Gamma Knife surgery for vestibular schwannomas

    Science.gov (United States)

    Lonneville, Sarah; Delbrouck, Carine; Renier, Cécile; Devriendt, Daniel; Massager, Nicolas

    2015-01-01

    Background: Gamma Knife (GK) surgery is a recognized treatment option for the management of small to medium-sized vestibular schwannoma (VS) associated with high-tumor control and low morbidity. When a radiosurgical treatment fails to stop tumor growth, repeat GK surgery can be proposed in selected cases. Methods: A series of 27 GK retreatments was performed in 25 patients with VS; 2 patients underwent three procedures. The median time interval between GK treatments was 45 months. The median margin dose used for the first, second, and third GK treatments was 12 Gy, 12 Gy, and 14 Gy, respectively. Six patients (4 patients for the second irradiation and 2 patients for the third irradiation) with partial tumor regrowth were treated only on the growing part of the tumor using a median margin dose of 13 Gy. The median tumor volume was 0.9, 2.3, and 0.7 cc for the first, second, and third treatments, respectively. Stereotactic positron emission tomography (PET) guidance was used for dose planning in 6 cases. Results: Mean follow-up duration was 46 months (range 24–110). At the last follow-up, 85% of schwannomas were controlled. The tumor volume decreased, remained unchanged, or increased after retreatment in 15, 8, and 4 cases, respectively. Four patients had PET during follow-up, and all showed a significant metabolic decrease of the tumor. Hearing was not preserved after retreatment in any patients. New facial or trigeminal palsy did not occur after retreatment. Conclusions: Our results support the long-term efficacy and low morbidity of repeat GK treatment for selected patients with tumor growth after initial treatment. PMID:26500799

  10. Molecular insight into mitochondrial DNA depletion syndrome in two patients with novel mutations in the deoxyguanosine kinase and thymidine kinase 2 genes.

    Science.gov (United States)

    Wang, Liya; Limongelli, Anna; Vila, Maya R; Carrara, Franco; Zeviani, Massimo; Eriksson, Staffan

    2005-01-01

    Thymidine kinase 2 (TK2) and deoxyguanosine kinase (dGK) are the two key enzymes in mitochondrial DNA (mtDNA) precursor synthesis. Deficiencies in TK2 or dGK activity, due to genetic alteration, have been shown to cause tissue-specific depletion of mtDNA. In the case of TK2 deficiency, affected individuals suffer severe myopathy and, in the case of dGK deficiency, devastating liver or multi-systemic disease. Here, we report clinical and biochemical findings from two patients with mtDNA depletion syndrome. Patient A was a compound heterozygote carrying the previously reported T77M mutation and a novel mutation (R161K) in the TK2 gene. Patient B carried a novel mutation (L250S) in the dGK gene. The clinical symptoms of patient A included muscular weakness and exercise intolerance due to a severe mitochondrial myopathy associated with a 92% reduction in mtDNA. There was minimal involvement of other organs. Patient B suffered from rapidly progressive, early onset fatal liver failure associated with profoundly decreased mtDNA levels in liver and, to a lesser extent, in skeletal muscle. Site-directed mutagenesis was used to introduce the mutations detected in patients A and B into the TK2 and dGK cDNAs, respectively. We then characterized each of these recombinant enzymes. Catalytic activities of the three mutant enzymes were reduced to about 2-4% for TK2 and 0.5% for dGK as compared to the wild-type enzymes. Altered competition between dCyd and dThd was observed for the T77M mutant. The residual activities of the two mitochondrial enzymes correlated directly with disease development.

  11. TMG-123, a novel glucokinase activator, exerts durable effects on hyperglycemia without increasing triglyceride in diabetic animal models.

    Science.gov (United States)

    Tsumura, Yoshinori; Tsushima, Yu; Tamura, Azusa; Hasebe, Makiko; Kanou, Masanobu; Kato, Hirotsugu; Kobayashi, Tsunefumi

    2017-01-01

    Glucokinase (GK) plays a critical role for maintaining glucose homeostasis with regulating glucose uptake in liver and insulin secretion in pancreas. GK activators have been reported to decrease blood glucose levels in patients with type 2 diabetes mellitus. However, clinical development of GK activators has failed due to the loss of glucose-lowering effects and increased plasma triglyceride levels after chronic treatment. Here, we generated a novel GK activator, TMG-123, examined its in vitro and in vivo pharmacological characteristics, and evaluated its risks of aforementioned clinical issues. TMG-123 selectively activated GK enzyme activity without increasing Vmax. TMG-123 improved glucose tolerance without increasing plasma insulin levels in both insulin-deficient (Goto-Kakizaki rats) and insulin-resistant (db/db mice) models. The beneficial effect on glucose tolerance was greater than results observed with clinically available antidiabetic drugs such as metformin and glibenclamide in Zucker Diabetic Fatty rats. TMG-123 also improved glucose tolerance in combination with metformin. After 4 weeks of administration, TMG-123 reduced the Hemoglobin A1c levels without affecting liver and plasma triglyceride levels in Goto-Kakizaki rats and Diet-Induced Obesity mice. Moreover, TMG-123 sustained its effect on Hemoglobin A1c levels even after 24 weeks of administration without affecting triglycerides. Taken together, these data indicate that TMG-123 exerts glucose-lowering effects in both insulin-deficient and -resistant diabetes, and sustains reduced Hemoglobin A1c levels without affecting hepatic and plasma triglycerides even after chronic treatment. Therefore, TMG-123 is expected to be an antidiabetic drug that overcomes the concerns previously reported with other GK activators.

  12. Genomic and metabolic disposition of non-obese type 2 diabetic rats to increased myocardial fatty acid metabolism.

    Directory of Open Access Journals (Sweden)

    Sriram Devanathan

    Full Text Available Lipotoxicity of the heart has been implicated as a leading cause of morbidity in Type 2 Diabetes Mellitus (T2DM. While numerous reports have demonstrated increased myocardial fatty acid (FA utilization in obese T2DM animal models, this diabetic phenotype has yet to be demonstrated in non-obese animal models of T2DM. Therefore, the present study investigates functional, metabolic, and genomic differences in myocardial FA metabolism in non-obese type 2 diabetic rats. The study utilized Goto-Kakizaki (GK rats at the age of 24 weeks. Each rat was imaged with small animal positron emission tomography (PET to estimate myocardial blood flow (MBF and myocardial FA metabolism. Echocardiograms (ECHOs were performed to assess cardiac function. Levels of triglycerides (TG and non-esterified fatty acids (NEFA were measured in both plasma and cardiac tissues. Finally, expression profiles for 168 genes that have been implicated in diabetes and FA metabolism were measured using quantitative PCR (qPCR arrays. GK rats exhibited increased NEFA and TG in both plasma and cardiac tissue. Quantitative PET imaging suggests that GK rats have increased FA metabolism. ECHO data indicates that GK rats have a significant increase in left ventricle mass index (LVMI and decrease in peak early diastolic mitral annular velocity (E' compared to Wistar rats, suggesting structural remodeling and impaired diastolic function. Of the 84 genes in each the diabetes and FA metabolism arrays, 17 genes in the diabetes array and 41 genes in the FA metabolism array were significantly up-regulated in GK rats. Our data suggest that GK rats' exhibit increased genomic disposition to FA and TG metabolism independent of obesity.

  13. High resolution structure of the ba3 cytochrome c oxidase from Thermus thermophilus in a lipidic environment.

    Directory of Open Access Journals (Sweden)

    Theresa Tiefenbrunn

    Full Text Available The fundamental chemistry underpinning aerobic life on Earth involves reduction of dioxygen to water with concomitant proton translocation. This process is catalyzed by members of the heme-copper oxidase (HCO superfamily. Despite the availability of crystal structures for all types of HCO, the mode of action for this enzyme is not understood at the atomic level, namely how vectorial H(+ and e(- transport are coupled. Toward addressing this problem, we report wild type and A120F mutant structures of the ba(3-type cytochrome c oxidase from Thermus thermophilus at 1.8 Å resolution. The enzyme has been crystallized from the lipidic cubic phase, which mimics the biological membrane environment. The structures reveal 20 ordered lipid molecules that occupy binding sites on the protein surface or mediate crystal packing interfaces. The interior of the protein encloses 53 water molecules, including 3 trapped in the designated K-path of proton transfer and 8 in a cluster seen also in A-type enzymes that likely functions in egress of product water and proton translocation. The hydrophobic O(2-uptake channel, connecting the active site to the lipid bilayer, contains a single water molecule nearest the Cu(B atom but otherwise exhibits no residual electron density. The active site contains strong electron density for a pair of bonded atoms bridging the heme Fe(a3 and Cu(B atoms that is best modeled as peroxide. The structure of ba(3-oxidase reveals new information about the positioning of the enzyme within the membrane and the nature of its interactions with lipid molecules. The atomic resolution details provide insight into the mechanisms of electron transfer, oxygen diffusion into the active site, reduction of oxygen to water, and pumping of protons across the membrane. The development of a robust system for production of ba(3-oxidase crystals diffracting to high resolution, together with an established expression system for generating mutants, opens the

  14. Amedofu, GK

    African Journals Online (AJOL)

    Characteristics of hearing-impairment among patients in Ghana Abstract PDF · Vol 14, No 1 (2007) - Articles Effectiveness of hearing conservation program at a large surface gold mining company in Ghana. Abstract PDF · Vol 14, No 3 (2007) - Articles Visual damage following direct sighting of solar eclipse in Ghana

  15. Characterization of DNA polymerase X from Thermus thermophilus HB8 reveals the POLXc and PHP domains are both required for 3'-5' exonuclease activity.

    Science.gov (United States)

    Nakane, Shuhei; Nakagawa, Noriko; Kuramitsu, Seiki; Masui, Ryoji

    2009-04-01

    The X-family DNA polymerases (PolXs) comprise a highly conserved DNA polymerase family found in all kingdoms. Mammalian PolXs are known to be involved in several DNA-processing pathways including repair, but the cellular functions of bacterial PolXs are less known. Many bacterial PolXs have a polymerase and histidinol phosphatase (PHP) domain at their C-termini in addition to a PolX core (POLXc) domain, and possess 3'-5' exonuclease activity. Although both domains are highly conserved in bacteria, their molecular functions, especially for a PHP domain, are unknown. We found Thermus thermophilus HB8 PolX (ttPolX) has Mg(2+)/Mn(2+)-dependent DNA/RNA polymerase, Mn(2+)-dependent 3'-5' exonuclease and DNA-binding activities. We identified the domains of ttPolX by limited proteolysis and characterized their biochemical activities. The POLXc domain was responsible for the polymerase and DNA-binding activities but exonuclease activity was not detected for either domain. However, the POLXc and PHP domains interacted with each other and a mixture of the two domains had Mn(2+)-dependent 3'-5' exonuclease activity. Moreover, site-directed mutagenesis revealed catalytically important residues in the PHP domain for the 3'-5' exonuclease activity. Our findings provide a molecular insight into the functional domain organization of bacterial PolXs, especially the requirement of the PHP domain for 3'-5' exonuclease activity.

  16. Models for the a subunits of the Thermus thermophilus V/A-ATPase and Saccharomyces cerevisiae V-ATPase enzymes by cryo-EM and evolutionary covariance

    Science.gov (United States)

    Schep, Daniel G.; Rubinstein, John L.

    2016-01-01

    Rotary ATPases couple ATP synthesis or hydrolysis to proton translocation across a membrane. However, understanding proton translocation has been hampered by a lack of structural information for the membrane-embedded a subunit. The V/A-ATPase from the eubacterium Thermus thermophilus is similar in structure to the eukaryotic V-ATPase but has a simpler subunit composition and functions in vivo to synthesize ATP rather than pump protons. We determined the T. thermophilus V/A-ATPase structure by cryo-EM at 6.4 Å resolution. Evolutionary covariance analysis allowed tracing of the a subunit sequence within the map, providing a complete model of the rotary ATPase. Comparing the membrane-embedded regions of the T. thermophilus V/A-ATPase and eukaryotic V-ATPase from Saccharomyces cerevisiae allowed identification of the α-helices that belong to the a subunit and revealed the existence of previously unknown subunits in the eukaryotic enzyme. Subsequent evolutionary covariance analysis enabled construction of a model of the a subunit in the S. cerevisae V-ATPase that explains numerous biochemical studies of that enzyme. Comparing the two a subunit structures determined here with a structure of the distantly related a subunit from the bovine F-type ATP synthase revealed a conserved pattern of residues, suggesting a common mechanism for proton transport in all rotary ATPases. PMID:26951669

  17. Mutations and phenotype in isolated glycerol kinase deficiency

    Energy Technology Data Exchange (ETDEWEB)

    Walker, A.P.; Muscatelli, F.; Stafford, A.N.; Monaco, A.P. [Inst. of Molecular Medicine, Oxford (United Kingdom)] [and others

    1996-06-01

    We demonstrate that isolated glycerol kinase (GK) deficiency in three families results from mutation of the Xp21 GK gene. GK mutations were detected in four patients with widely differing phenotypes. Patient 1 had a splice-site mutation causing premature termination. His general health was good despite absent GK activity, indicating that isolated GK deficiency can be silent. Patient 2 had GK deficiency and a severe phenotype involving psychomotor retardation and growth delay, bone dysplasia, and seizures, similar to the severe phenotype of one of the first described cases of GK deficiency. His younger brother, patient 3, also had GK deficiency, but so far his development has been normal. GK exon 17 was deleted in both brothers, implicating additional factors in causation of the severe phenotype of patient 2. Patient 4 had both GK deficiency with mental retardation and a GK missense mutation (D440V). Possible explanations for the phenotypic variation of these four patients include ascertainment bias; metabolic or environmental stress as a precipitating factor in revealing GK-related changes, as has previously been described in juvenile GK deficiency; and interactions with functional polymorphisms in other genes that alter the effect of GK deficiency on normal development. 36 refs., 4 figs., 1 tab.

  18. Complete genome sequence of Truepera radiovictrix type strain (RQ-24).

    Science.gov (United States)

    Ivanova, Natalia; Rohde, Christine; Munk, Christine; Nolan, Matt; Lucas, Susan; Del Rio, Tijana Glavina; Tice, Hope; Deshpande, Shweta; Cheng, Jan-Fang; Tapia, Roxane; Han, Cliff; Goodwin, Lynne; Pitluck, Sam; Liolios, Konstantinos; Mavromatis, Konstantinos; Mikhailova, Natalia; Pati, Amrita; Chen, Amy; Palaniappan, Krishna; Land, Miriam; Hauser, Loren; Chang, Yun-Juan; Jeffries, Cynthia D; Brambilla, Evelyne; Rohde, Manfred; Göker, Markus; Tindall, Brian J; Woyke, Tanja; Bristow, James; Eisen, Jonathan A; Markowitz, Victor; Hugenholtz, Philip; Kyrpides, Nikos C; Klenk, Hans-Peter; Lapidus, Alla

    2011-02-22

    Truepera radiovictrix Albuquerque et al. 2005 is the type species of the genus Truepera within the phylum "Deinococcus/Thermus". T. radiovictrix is of special interest not only because of its isolated phylogenetic location in the order Deinococcales, but also because of its ability to grow under multiple extreme conditions in alkaline, moderately saline, and high temperature habitats. Of particular interest is the fact that, T. radiovictrix is also remarkably resistant to ionizing radiation, a feature it shares with members of the genus Deinococcus. This is the first completed genome sequence of a member of the family Trueperaceae and the fourth type strain genome sequence from a member of the order Deinococcales. The 3,260,398 bp long genome with its 2,994 protein-coding and 52 RNA genes consists of one circular chromosome and is a part of the Genomic Encyclopedia of Bacteria and Archaea project.

  19. Reaction of cyanide with cytochrome ba3 from Thermus thermophilus: spectroscopic characterization of the Fe(II)a3-CN.Cu(II)B-CN complex suggests four 14N atoms are coordinated to CuB.

    Science.gov (United States)

    Surerus, K K; Oertling, W A; Fan, C; Gurbiel, R J; Einarsdóttir, O; Antholine, W E; Dyer, R B; Hoffman, B M; Woodruff, W H; Fee, J A

    1992-01-01

    Cytochrome ba3 from Thermus thermophilus reacts slowly with excess HCN at pH 7.4 to create a form of the enzyme in which CuA, cytochrome b, and CuB remain oxidized, while cytochrome a3 is reduced by one electron, presumably with the formation of cyanogen. We have examined this form of the enzyme by UV-visible, resonance Raman, EPR, and electron nuclear double resonance spectroscopies in conjunction with permutations of 13C- and 15N-labeled cyanide. The results support a model in which one CN- binds through the carbon atom to ferrous a3, supporting a low-spin (S = 0) configuration on the Fe; bridging by this cyanide to the CuB is weak or absent. Four 14N atoms, presumably donated by histidine residues of the protein, provide a strong equatorial ligand field about CuB; a second CN- is coordinated through the carbon atom to CuB in an axial position. PMID:1314380

  20. NCBI nr-aa BLAST: CBRC-TBEL-01-0397 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TBEL-01-0397 ref|YP_145234.1| competence protein ComEC [Thermus thermophilus HB8] dbj|BAD71791.1| compe...tence protein ComEC [Thermus thermophilus HB8] YP_145234.1 0.020 31% ...

  1. A comparison of structural and evolutionary attributes of Escherichia coli and Thermus thermophilus small ribosomal subunits: signatures of thermal adaptation.

    Directory of Open Access Journals (Sweden)

    Saurav Mallik

    Full Text Available Here we compare the structural and evolutionary attributes of Thermus thermophilus and Escherichia coli small ribosomal subunits (SSU. Our results indicate that with few exceptions, thermophilic 16S ribosomal RNA (16S rRNA is densely packed compared to that of mesophilic at most of the analogous spatial regions. In addition, we have located species-specific cavity clusters (SSCCs in both species. E. coli SSCCs are numerous and larger compared to T. thermophilus SSCCs, which again indicates densely packed thermophilic 16S rRNA. Thermophilic ribosomal proteins (r-proteins have longer disordered regions than their mesophilic homologs and they experience larger disorder-to-order transitions during SSU-assembly. This is reflected in the predicted higher conformational changes of thermophilic r-proteins compared to their mesophilic homologs during SSU-assembly. This high conformational change of thermophilic r-proteins may help them to associate with the 16S ribosomal RNA with high complementary interfaces, larger interface areas, and denser molecular contacts, compared to those of mesophilic. Thus, thermophilic protein-rRNA interfaces are tightly associated with 16S rRNA than their mesophilic homologs. Densely packed 16S rRNA interior and tight protein-rRNA binding of T. thermophilus (compared to those of E. coli are likely the signatures of its thermal adaptation. We have found a linear correlation between the free energy of protein-RNA interface formation, interface size, and square of conformational changes, which is followed in both prokaryotic and eukaryotic SSU. Disorder is associated with high protein-RNA interface polarity. We have found an evolutionary tendency to maintain high polarity (thereby disorder at protein-rRNA interfaces, than that at rest of the protein structures. However, some proteins exhibit exceptions to this general trend.

  2. Structures of a putative RNA 5-methyluridine methyltransferase, Thermus thermophilus TTHA1280, and its complex with S-adenosyl-l-homocysteine

    International Nuclear Information System (INIS)

    Pioszak, Augen A.; Murayama, Kazutaka; Nakagawa, Noriko; Ebihara, Akio; Kuramitsu, Seiki; Shirouzu, Mikako; Yokoyama, Shigeyuki

    2005-01-01

    Three structures of a putative RNA 5-methyluridine methyltransferase from T. thermophilus, including its complex with S-adenosyl-l-homocysteine, are presented. The structures reveal the mode of cofactor binding, architecture of the putative active site, and the presence of a deep cleft adjacent to the active site that may bind RNA. The Thermus thermophilus hypothetical protein TTHA1280 belongs to a family of predicted S-adenosyl-l-methionine (AdoMet) dependent RNA methyltransferases (MTases) present in many bacterial and archaeal species. Inspection of amino-acid sequence motifs common to class I Rossmann-fold-like MTases suggested a specific role as an RNA 5-methyluridine MTase. Selenomethionine (SeMet) labelled and native versions of the protein were expressed, purified and crystallized. Two crystal forms of the SeMet-labelled apoprotein were obtained: SeMet-ApoI and SeMet-ApoII. Cocrystallization of the native protein with S-adenosyl-l-homocysteine (AdoHcy) yielded a third crystal form, Native-AdoHcy. The SeMet-ApoI structure was solved by the multiple anomalous dispersion method and refined at 2.55 Å resolution. The SeMet-ApoII and Native-AdoHcy structures were solved by molecular replacement and refined at 1.80 and 2.60 Å, respectively. TTHA1280 formed a homodimer in the crystals and in solution. Each subunit folds into a three-domain structure composed of a small N-terminal PUA domain, a central α/β-domain and a C-terminal Rossmann-fold-like MTase domain. The three domains form an overall clamp-like shape, with the putative active site facing a deep cleft. The architecture of the active site is consistent with specific recognition of uridine and catalysis of methyl transfer to the 5-carbon position. The cleft is suitable in size and charge distribution for binding single-stranded RNA.

  3. Divergent Immunomodulation Capacity of Individual Myelin Peptides—Components of Liposomal Therapeutic against Multiple Sclerosis

    Directory of Open Access Journals (Sweden)

    Vilena V. Ivanova

    2017-10-01

    Full Text Available Multiple sclerosis (MS is an autoimmune disease characterized by demyelination and consequent neuron injury. Although the pathogenesis of MS is largely unknown, a breach in immune self-tolerance to myelin followed by development of autoreactive encephalitogenic T cells is suggested to play the central role. The myelin basic protein (MBP is believed to be one of the main targets for autoreactive lymphocytes. Recently, immunodominant MBP peptides encapsulated into the mannosylated liposomes, referred as Xemys, were shown to suppress development of experimental autoimmune encephalomyelitis, a rodent model of MS, and furthermore passed the initial stage of clinical trials. Here, we investigated the role of individual polypeptide components [MBP peptides 46–62 (GH17, 124–139 (GK16, and 147–170 (QR24] of this liposomal peptide therapeutic in cytokine release and activation of immune cells from MS patients and healthy donors. The overall effects were assessed using peripheral blood mononuclear cells (PBMCs, whereas alterations in antigen-presenting capacities were studied utilizing plasmacytoid dendritic cells (pDCs. Among three MBP-immunodominant peptides, QR24 and GK16 activated leukocytes, while GH17 was characterized by an immunosuppressive effect. Peptides QR24 and GK16 upregulated CD4 over CD8 T cells and induced proliferation of CD25+ cells, whereas GH17 decreased the CD4/CD8 T cell ratio and had limited effects on CD25+ T cells. Accordingly, components of liposomal peptide therapeutic differed in upregulation of cytokines upon addition to PBMCs and pDCs. Peptide QR24 was evidently more effective in upregulation of pro-inflammatory cytokines, whereas GH17 significantly increased production of IL-10 through treated cells. Altogether, these data suggest a complexity of action of the liposomal peptide therapeutic that does not seem to involve simple helper T cells (Th-shift but rather the rebalancing of the immune system.

  4. Functional expression of a penicillin acylase from the extreme thermophile Thermus thermophilus HB27 in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Torres Leticia L

    2012-08-01

    Full Text Available Abstract Background Penicillin acylases (PACs are enzymes of industrial relevance in the manufacture of β-lactam antibiotics. Development of a PAC with a longer half-life under the reaction conditions used is essential for the improvement of the operational stability of the process. A gene encoding a homologue to Escherichia coli PAC was found in the genome of the thermophilic bacterium Thermus thermophilus (Tth HB27. Because of the nature of this PAC and its complex maturation that is crucial to reach its functional heterodimeric final conformation, the overexpression of this enzyme in a heterologous mesophilic host was a challenge. Here we describe the purification and characterization of the PAC protein from Tth HB27 overexpressed in Escherichia coli. Results Fusions to a superfolder green fluorescent protein and differential membrane solubilization assays indicated that the native enzyme remains attached through its amino-terminal end to the outer side of the cytoplasmic membrane of Tth cells. In order to overexpress this PAC in E. coli cells, a variant of the protein devoid of its membrane anchoring segment was constructed. The effect of the co-expression of chaperones and calcium supplementation of the culture medium was investigated. The total production of PAC was enhanced by the presence of DnaK/J and GrpE and even more by trigger factor and GroEL/ES. In addition, 10 mM calcium markedly improved both PAC specific and volumetric activities. Recombinant PAC was affinity-purified and proper maturation of the protein was confirmed by SDS-PAGE and MALDI-TOF analysis of the subunits. The recombinant protein was tested for activity towards several penicillins, cephalosporins and homoserine lactones. Hydrophobic acyl-chain penicillins were preferred over the rest of the substrates. Penicillin K (octanoyl penicillin was the best substrate, with the highest specificity constant value (16.12 mM-1.seg-1. The optimum pH was aprox. 4 and the optimum

  5. Functional expression of a penicillin acylase from the extreme thermophile Thermus thermophilus HB27 in Escherichia coli.

    Science.gov (United States)

    Torres, Leticia L; Ferreras, Eloy R; Cantero, Angel; Hidalgo, Aurelio; Berenguer, José

    2012-08-09

    Penicillin acylases (PACs) are enzymes of industrial relevance in the manufacture of β-lactam antibiotics. Development of a PAC with a longer half-life under the reaction conditions used is essential for the improvement of the operational stability of the process. A gene encoding a homologue to Escherichia coli PAC was found in the genome of the thermophilic bacterium Thermus thermophilus (Tth) HB27. Because of the nature of this PAC and its complex maturation that is crucial to reach its functional heterodimeric final conformation, the overexpression of this enzyme in a heterologous mesophilic host was a challenge. Here we describe the purification and characterization of the PAC protein from Tth HB27 overexpressed in Escherichia coli. Fusions to a superfolder green fluorescent protein and differential membrane solubilization assays indicated that the native enzyme remains attached through its amino-terminal end to the outer side of the cytoplasmic membrane of Tth cells. In order to overexpress this PAC in E. coli cells, a variant of the protein devoid of its membrane anchoring segment was constructed. The effect of the co-expression of chaperones and calcium supplementation of the culture medium was investigated. The total production of PAC was enhanced by the presence of DnaK/J and GrpE and even more by trigger factor and GroEL/ES. In addition, 10 mM calcium markedly improved both PAC specific and volumetric activities. Recombinant PAC was affinity-purified and proper maturation of the protein was confirmed by SDS-PAGE and MALDI-TOF analysis of the subunits. The recombinant protein was tested for activity towards several penicillins, cephalosporins and homoserine lactones. Hydrophobic acyl-chain penicillins were preferred over the rest of the substrates. Penicillin K (octanoyl penicillin) was the best substrate, with the highest specificity constant value (16.12 mM-1.seg-1). The optimum pH was aprox. 4 and the optimum temperature was 75 °C. The half-life of

  6. Complete genome sequence of Truepera radiovictrix type strain (RQ-24T)

    Energy Technology Data Exchange (ETDEWEB)

    Ivanova, N [U.S. Department of Energy, Joint Genome Institute; Rohde, Christine [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Munk, Christine [Joint Genome Institute, Walnut Creek, California; Nolan, Matt [Joint Genome Institute, Walnut Creek, California; Lucas, Susan [Joint Genome Institute, Walnut Creek, California; Glavina Del Rio, Tijana [Joint Genome Institute, Walnut Creek, California; Tice, Hope [Joint Genome Institute, Walnut Creek, California; Deshpande, Shweta [Joint Genome Institute, Walnut Creek, California; Cheng, Jan-Fang [Joint Genome Institute, Walnut Creek, California; Tapia, Roxanne [Los Alamos National Laboratory (LANL); Han, Cliff [Los Alamos National Laboratory (LANL); Goodwin, Lynne A. [Los Alamos National Laboratory (LANL); Pitluck, Sam [Joint Genome Institute, Walnut Creek, California; Liolios, Konstantinos [Joint Genome Institute, Walnut Creek, California; Mavromatis, K [U.S. Department of Energy, Joint Genome Institute; Pati, Amrita [U.S. Department of Energy, Joint Genome Institute; Chen, Amy [Joint Genome Institute, Walnut Creek, California; Palaniappan, Krishna [Joint Genome Institute, Walnut Creek, California; Land, Miriam L [ORNL; Hauser, Loren John [ORNL; Chang, Yun-Juan [ORNL; Jeffries, Cynthia [Oak Ridge National Laboratory (ORNL); Brambilla, Evelyne-Marie [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Rohde, Manfred [HZI - Helmholtz Centre for Infection Research, Braunschweig, Germany; Goker, Markus [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Tindall, Brian [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Woyke, Tanja [Joint Genome Institute, Walnut Creek, California; Bristow, James [Joint Genome Institute, Walnut Creek, California; Eisen, Jonathan [Joint Genome Institute, Walnut Creek, California; Markowitz, Victor [Joint Genome Institute, Walnut Creek, California; Hugenholtz, Philip [U.S. Department of Energy, Joint Genome Institute; Kyrpides, Nikos C [Joint Genome Institute, Walnut Creek, California; Klenk, Hans-Peter [DSMZ - German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany; Lapidus, Alla L. [Joint Genome Institute, Walnut Creek, California

    2011-01-01

    Truepera radiovictrix Albuquerque et al. 2005 is the type species of the genus Truepera within the phylum Deinococcus/Thermus. T. radiovictrix is of special interest not only because of its isolated phylogenetic location in the order Deinococcales, but also because of its ability to grow under multiple extreme conditions in alkaline, moderately saline, and high temperature habitats. Of particular interest is the fact that, T. radiovictrix is also remarkably resistant to ionizing radiation, a feature it shares with members of the genus Deinococcus. This is the first completed genome sequence of a member of the family Trueperaceae and the fourth type strain genome sequence from a member of the order Deinococcales. The 3,260,398 bp long genome with its 2,994 protein-coding and 52 RNA genes consists of one circular chromosome and is a part of the Genomic Encyclopedia of Bacteria and Archaea project.

  7. Susceptibility of glucokinase-MODY mutants to inactivation by oxidative stress in pancreatic β-cells.

    Science.gov (United States)

    Cullen, Kirsty S; Matschinsky, Franz M; Agius, Loranne; Arden, Catherine

    2011-12-01

    The posttranslational regulation of glucokinase (GK) differs in hepatocytes and pancreatic β-cells. We tested the hypothesis that GK mutants that cause maturity-onset diabetes of the young (GK-MODY) show compromised activity and posttranslational regulation in β-cells. Activity and protein expression of GK-MODY and persistent hyperinsulinemic hypoglycemia of infancy (PHHI) mutants were studied in β-cell (MIN6) and non-β-cell (H4IIE) models. Binding of GK to phosphofructo-2-kinase, fructose-2,6-bisphosphatase (PFK2/FBPase2) was studied by bimolecular fluorescence complementation in cell-based models. Nine of 11 GK-MODY mutants that have minimal effect on enzyme kinetics in vitro showed decreased specific activity relative to wild type when expressed in β-cells. A subset of these were stable in non-β-cells but showed increased inactivation in conditions of oxidative stress and partial reversal of inactivation by dithiothreitol. Unlike the GK-MODY mutants, four of five GK-PHHI mutants had similar specific activity to wild type and Y214C had higher activity than wild type. The GK-binding protein PFK2/FBPase2 protected wild-type GK from oxidative inactivation and the decreased stability of GK-MODY mutants correlated with decreased interaction with PFK2/FBPase2. Several GK-MODY mutants show posttranslational defects in β-cells characterized by increased susceptibility to oxidative stress and/or protein instability. Regulation of GK activity through modulation of thiol status may be a physiological regulatory mechanism for the control of GK activity in β-cells.

  8. Enhancement of lymphocyte proliferation by mouse glandular kallikrein.

    Science.gov (United States)

    Hu, Z Q; Murakami, K; Ikigai, H; Shimamura, T

    1992-03-01

    Mouse glandular kallikrein (mGK) strongly enhanced the spontaneous and mitogen-induced proliferation of lymphocytes. Both blast formation and 3H-TdR incorporation were dose-dependently enhanced at the same time many cells were killed. The enhancing activity was independent of EGF, because EGF-binding proteins (mGK-9 in mGK-6,9 mixture and mGK-13), renal kallikrein (mGK-6) and human kallikrein all displayed the same enhancement. A serine proteinase inhibitor, diisopropyl fluorophosphate, could block the enhancement by mGK. The new function suggests that mGK is important in the immune system as a regulatory molecule.

  9. Adaptive fractionated stereotactic Gamma Knife radiotherapy of meningioma using integrated stereotactic cone-beam-CT and adaptive re-planning (a-gkFSRT)

    Energy Technology Data Exchange (ETDEWEB)

    Stieler, F.; Wenz, F.; Abo-Madyan, Y.; Schweizer, B.; Polednik, M.; Herskind, C.; Giordano, F.A.; Mai, S. [University of Heidelberg, Department of Radiation Oncology, University Medical Center Mannheim, Mannheim (Germany)

    2016-11-15

    The Gamma Knife Icon (Elekta AB, Stockholm, Sweden) allows frameless stereotactic treatment using a combination of cone beam computer tomography (CBCT), a thermoplastic mask system, and an infrared-based high-definition motion management (HDMM) camera system for patient tracking during treatment. We report on the first patient with meningioma at the left petrous bone treated with adaptive fractionated stereotactic radiotherapy (a-gkFSRT). The first patient treated with Gamma Knife Icon at our institute received MR imaging for preplanning before treatment. For each treatment fraction, a daily CBCT was performed to verify the actual scull/tumor position. The system automatically adapted the planned shot positions to the daily position and recalculated the dose distribution (online adaptive planning). During treatment, the HDMM system recorded the intrafractional patient motion. Furthermore, the required times were recorded to define a clinical treatment slot. Total treatment time was around 20 min. Patient positioning needed 0.8 min, CBCT positioning plus acquisition 1.65 min, CT data processing and adaptive planning 2.66 min, and treatment 15.6 min. The differences for the five daily CBCTs compared to the reference are for rotation: -0.59 ± 0.49 /0.18 ± 0.20 /0.05 ± 0.36 and for translation: 0.94 ± 0.52 mm/-0.08 ± 0.08 mm/-1.13 ± 0.89 mm. Over all fractions, an intrafractional movement of 0.13 ± 0.04 mm was observed. The Gamma Knife Icon allows combining the accuracy of the stereotactic Gamma Knife system with the flexibility of fractionated treatment with the mask system and CBCT. Furthermore, the Icon system introduces a new online patient tracking system to the clinical routine. The interfractional accuracy of patient positioning was controlled with a thermoplastic mask and CBCT. (orig.) [German] Das Gamma Knife Icon (Elekta AB, Stockholm, Schweden) ermoeglicht die stereotaktische Behandlung von Patienten mittels Cone-beam-Computertomographie (CBCT

  10. Gamma knife irradiation of injured sciatic nerve induces histological and behavioral improvement in the rat neuropathic pain model.

    Directory of Open Access Journals (Sweden)

    Yuki Yagasaki

    Full Text Available We examined the effects of gamma knife (GK irradiation on injured nerves using a rat partial sciatic nerve ligation (PSL model. GK irradiation was performed at one week after ligation and nerve preparations were made three weeks after ligation. GK irradiation is known to induce immune responses such as glial cell activation in the central nervous system. Thus, we determined the effects of GK irradiation on macrophages using immunoblot and histochemical analyses. Expression of Iba-1 protein, a macrophage marker, was further increased in GK-treated injured nerves as compared with non-irradiated injured nerves. Immunohistochemical study of Iba-1 in GK-irradiated injured sciatic nerves demonstrated Iba-1 positive macrophage accumulation to be enhanced in areas distal to the ligation point. In the same area, myelin debris was also more efficiently removed by GK-irradiation. Myelin debris clearance by macrophages is thought to contribute to a permissive environment for axon growth. In the immunoblot study, GK irradiation significantly increased expressions of βIII-tubulin protein and myelin protein zero, which are markers of axon regeneration and re-myelination, respectively. Toluidine blue staining revealed the re-myelinated fiber diameter to be larger at proximal sites and that the re-myelinated fiber number was increased at distal sites in GK-irradiated injured nerves as compared with non-irradiated injured nerves. These results suggest that GK irradiation of injured nerves facilitates regeneration and re-myelination. In a behavior study, early alleviation of allodynia was observed with GK irradiation in PSL rats. When GK-induced alleviation of allodynia was initially detected, the expression of glial cell line-derived neurotrophic factor (GDNF, a potent analgesic factor, was significantly increased by GK irradiation. These results suggested that GK irradiation alleviates allodynia via increased GDNF. This study provides novel evidence that GK

  11. Analysis of the cooperative ATPase cycle of the AAA+ chaperone ClpB from Thermus thermophilus by using ordered heterohexamers with an alternating subunit arrangement.

    Science.gov (United States)

    Yamasaki, Takashi; Oohata, Yukiko; Nakamura, Toshiki; Watanabe, Yo-hei

    2015-04-10

    The ClpB/Hsp104 chaperone solubilizes and reactivates protein aggregates in cooperation with DnaK/Hsp70 and its cofactors. The ClpB/Hsp104 protomer has two AAA+ modules, AAA-1 and AAA-2, and forms a homohexamer. In the hexamer, these modules form a two-tiered ring in which each tier consists of homotypic AAA+ modules. By ATP binding and its hydrolysis at these AAA+ modules, ClpB/Hsp104 exerts the mechanical power required for protein disaggregation. Although ATPase cycle of this chaperone has been studied by several groups, an integrated understanding of this cycle has not been obtained because of the complexity of the mechanism and differences between species. To improve our understanding of the ATPase cycle, we prepared many ordered heterohexamers of ClpB from Thermus thermophilus, in which two subunits having different mutations were cross-linked to each other and arranged alternately and measured their nucleotide binding, ATP hydrolysis, and disaggregation abilities. The results indicated that the ATPase cycle of ClpB proceeded as follows: (i) the 12 AAA+ modules randomly bound ATP, (ii) the binding of four or more ATP to one AAA+ ring was sensed by a conserved Arg residue and converted another AAA+ ring into the ATPase-active form, and (iii) ATP hydrolysis occurred cooperatively in each ring. We also found that cooperative ATP hydrolysis in at least one ring was needed for the disaggregation activity of ClpB. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  12. NCBI nr-aa BLAST: CBRC-MLUC-01-1078 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MLUC-01-1078 ref|ZP_03497741.1| DNA internalization-related competence protein... ComEC/Rec2 [Thermus aquaticus Y51MC23] gb|EED09156.1| DNA internalization-related competence protein ComEC/Rec2 [Thermus aquaticus Y51MC23] ZP_03497741.1 0.005 32% ...

  13. Antidiabetic effects of glucokinase regulatory protein small-molecule disruptors

    Science.gov (United States)

    Lloyd, David J.; St Jean, David J.; Kurzeja, Robert J. M.; Wahl, Robert C.; Michelsen, Klaus; Cupples, Rod; Chen, Michelle; Wu, John; Sivits, Glenn; Helmering, Joan; Komorowski, Renée; Ashton, Kate S.; Pennington, Lewis D.; Fotsch, Christopher; Vazir, Mukta; Chen, Kui; Chmait, Samer; Zhang, Jiandong; Liu, Longbin; Norman, Mark H.; Andrews, Kristin L.; Bartberger, Michael D.; van, Gwyneth; Galbreath, Elizabeth J.; Vonderfecht, Steven L.; Wang, Minghan; Jordan, Steven R.; Véniant, Murielle M.; Hale, Clarence

    2013-12-01

    Glucose homeostasis is a vital and complex process, and its disruption can cause hyperglycaemia and type II diabetes mellitus. Glucokinase (GK), a key enzyme that regulates glucose homeostasis, converts glucose to glucose-6-phosphate in pancreatic β-cells, liver hepatocytes, specific hypothalamic neurons, and gut enterocytes. In hepatocytes, GK regulates glucose uptake and glycogen synthesis, suppresses glucose production, and is subject to the endogenous inhibitor GK regulatory protein (GKRP). During fasting, GKRP binds, inactivates and sequesters GK in the nucleus, which removes GK from the gluconeogenic process and prevents a futile cycle of glucose phosphorylation. Compounds that directly hyperactivate GK (GK activators) lower blood glucose levels and are being evaluated clinically as potential therapeutics for the treatment of type II diabetes mellitus. However, initial reports indicate that an increased risk of hypoglycaemia is associated with some GK activators. To mitigate the risk of hypoglycaemia, we sought to increase GK activity by blocking GKRP. Here we describe the identification of two potent small-molecule GK-GKRP disruptors (AMG-1694 and AMG-3969) that normalized blood glucose levels in several rodent models of diabetes. These compounds potently reversed the inhibitory effect of GKRP on GK activity and promoted GK translocation both in vitro (isolated hepatocytes) and in vivo (liver). A co-crystal structure of full-length human GKRP in complex with AMG-1694 revealed a previously unknown binding pocket in GKRP distinct from that of the phosphofructose-binding site. Furthermore, with AMG-1694 and AMG-3969 (but not GK activators), blood glucose lowering was restricted to diabetic and not normoglycaemic animals. These findings exploit a new cellular mechanism for lowering blood glucose levels with reduced potential for hypoglycaemic risk in patients with type II diabetes mellitus.

  14. Browse Title Index - African Journals Online

    African Journals Online (AJOL)

    Items 601 - 650 of 1038 ... Vol 24 (2002), Note on an invasion of intertidal zoanthid colonies by a chaetopterid polychaete at Park Rynie Beach, KwaZulu-Natal, South Africa, Abstract PDF. R Biseswas, GK Moodley, AD Naidoo. Vol 38, No 4 (2016), Nutrient limitation in two marine diatoms inhibits release of bromoform, Abstract.

  15. A Key Enzyme of the NAD+ Salvage Pathway in Thermus thermophilus: Characterization of Nicotinamidase and the Impact of Its Gene Deletion at High Temperatures.

    Science.gov (United States)

    Taniguchi, Hironori; Sungwallek, Sathidaphorn; Chotchuang, Phatcharin; Okano, Kenji; Honda, Kohsuke

    2017-09-01

    NAD (NAD + ) is a cofactor related to many cellular processes. This cofactor is known to be unstable, especially at high temperatures, where it chemically decomposes to nicotinamide and ADP-ribose. Bacteria, yeast, and higher organisms possess the salvage pathway for reconstructing NAD + from these decomposition products; however, the importance of the salvage pathway for survival is not well elucidated, except for in pathogens lacking the NAD + de novo synthesis pathway. Herein, we report the importance of the NAD + salvage pathway in the thermophilic bacterium Thermus thermophilus HB8 at high temperatures. We identified the gene encoding nicotinamidase (TTHA0328), which catalyzes the first reaction of the NAD + salvage pathway. This recombinant enzyme has a high catalytic activity against nicotinamide ( K m of 17 μM, k cat of 50 s -1 , k cat / K m of 3.0 × 10 3 s -1 · mM -1 ). Deletion of this gene abolished nicotinamide deamination activity in crude extracts of T. thermophilus and disrupted the NAD + salvage pathway in T. thermophilus Disruption of the salvage pathway led to the severe growth retardation at a higher temperature (80°C), owing to the drastic decrease in the intracellular concentrations of NAD + and NADH. IMPORTANCE NAD + and other nicotinamide cofactors are essential for cell metabolism. These molecules are unstable and decompose, even under the physiological conditions in most organisms. Thermophiles can survive at high temperatures where NAD + decomposition is, in general, more rapid. This study emphasizes that NAD + instability and its homeostasis can be one of the important factors for thermophile survival in extreme temperatures. Copyright © 2017 American Society for Microbiology.

  16. The use of fluorescent intrabodies to detect endogenous gankyrin in living cancer cells

    International Nuclear Information System (INIS)

    Rinaldi, Anne-Sophie; Freund, Guillaume; Desplancq, Dominique; Sibler, Annie-Paule; Baltzinger, Mireille; Rochel, Natacha; Mély, Yves; Didier, Pascal; Weiss, Etienne

    2013-01-01

    Expression of antibody fragments in mammalian cells (intrabodies) is used to probe the target protein or interfere with its biological function. We previously described the in vitro characterisation of a single-chain Fv (scFv) antibody fragment (F5) isolated from an intrabody library that binds to the oncoprotein gankyrin (GK) in solution. Here, we have isolated several other scFvs that interact with GK in the presence of F5 and tested whether they allow, when fused to fluorescent proteins, to detect by FRET endogenous GK in living cells. The binding of pairs of scFvs to GK was analysed by gel filtration and the ability of each scFv to mediate nuclear import/export of GK was determined. Binding between scFv-EGFP and RFP-labelled GK in living cells was detected by fluorescence lifetime imaging microscopy (FLIM). After co-transfection of two scFvs fused to EGFP and RFP, respectively, which form a tri-molecular complex with GK in vitro, FRET signal was measured. This system allowed us to observe that GK is monomeric and distributed throughout the cytoplasm and nucleus of several cancer cell lines. Our results show that pairs of fluorescently labelled intrabodies can be monitored by FLIM–FRET microscopy and that this technique allows the detection of lowly expressed endogenous proteins in single living cells. Highlights: ► Endogenous GK in living cells was targeted with pairs of fluorescently-tagged scFvs. ► Tri-molecular complexes containing two scFvs and one molecule GK were formed. ► GK was detected using fluorescence lifetime-based FRET imaging. ► GK is monomeric and homogeneously distributed in several cancer cell lines. ► This technique may have many applications in live-cell imaging of endogenous proteins

  17. The use of fluorescent intrabodies to detect endogenous gankyrin in living cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Rinaldi, Anne-Sophie; Freund, Guillaume; Desplancq, Dominique; Sibler, Annie-Paule; Baltzinger, Mireille [Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch (France); Rochel, Natacha [Institut de Génétique et de Biologie Moléculaire et Cellulaire, UMR 7104, CNRS/INSERM/Université de Strasbourg, rue Laurent Fries, 67404 Illkirch (France); Mély, Yves; Didier, Pascal [Faculté de Pharmacie, UMR 7213, CNRS/Université de Strasbourg, route du Rhin, 67401 Illkirch (France); Weiss, Etienne, E-mail: eweiss@unistra.fr [Ecole Supérieure de Biotechnologie de Strasbourg, UMR 7242, CNRS/Université de Strasbourg, boulevard Sébastien Brant, 67412 Illkirch (France)

    2013-04-01

    Expression of antibody fragments in mammalian cells (intrabodies) is used to probe the target protein or interfere with its biological function. We previously described the in vitro characterisation of a single-chain Fv (scFv) antibody fragment (F5) isolated from an intrabody library that binds to the oncoprotein gankyrin (GK) in solution. Here, we have isolated several other scFvs that interact with GK in the presence of F5 and tested whether they allow, when fused to fluorescent proteins, to detect by FRET endogenous GK in living cells. The binding of pairs of scFvs to GK was analysed by gel filtration and the ability of each scFv to mediate nuclear import/export of GK was determined. Binding between scFv-EGFP and RFP-labelled GK in living cells was detected by fluorescence lifetime imaging microscopy (FLIM). After co-transfection of two scFvs fused to EGFP and RFP, respectively, which form a tri-molecular complex with GK in vitro, FRET signal was measured. This system allowed us to observe that GK is monomeric and distributed throughout the cytoplasm and nucleus of several cancer cell lines. Our results show that pairs of fluorescently labelled intrabodies can be monitored by FLIM–FRET microscopy and that this technique allows the detection of lowly expressed endogenous proteins in single living cells. Highlights: ► Endogenous GK in living cells was targeted with pairs of fluorescently-tagged scFvs. ► Tri-molecular complexes containing two scFvs and one molecule GK were formed. ► GK was detected using fluorescence lifetime-based FRET imaging. ► GK is monomeric and homogeneously distributed in several cancer cell lines. ► This technique may have many applications in live-cell imaging of endogenous proteins.

  18. Radioimmunological determination of glucokinase turnover in the rat liver

    International Nuclear Information System (INIS)

    Sibrowski, W.

    1980-01-01

    Synthesis and degradation of glucokinase (GK), key enzyme of glucose utilisation has been investigated in vivo in intact and adrenalectomized rats. Four statements can be made: a pure glucose diet stimulates GK synthesis, glucocorticoids have a permissive effect in the induction of GK sythesis, glucocorticoids do not have any effect in themselves, glucocorticoids affect GK degradation. (orig./AJ) [de

  19. [Glucokinase and glucokinase regulatory proteins as molecular targets for novel antidiabetic drugs].

    Science.gov (United States)

    Rubtsov, P M; Igudin, E L; Tiulpakov, A N

    2015-01-01

    The impairment of glucose homeostasis leads to hyperglycemia and type-2 diabetes mellitus. Glucokinase (GK), an enzyme that catalyzes the conversion of glucose to glucose-6-phosphate in pancreatic ß-cells, liver hepatocytes, specific hypothalamic neurons, and intestine enterocytes, is a key regulator of glucose homeostasis. In hepatocytes, GK controls the glucose uptake and glycogen synthesis and inhibits the glucose synthesis via the gluconeogenesis pathway. Glucokinase regulatory protein (GKRP) synthesized in hepatocytes acts as an endogenous GK inhibitor. During fasting, GKRP binds GK, inactivates it, and transports it into the cell nucleus, thus isolating it from the hepatocyte carbohydrate metabolism. In the beginning of the 2000s, the research was mainly focused on the development and trials of the small molecule GK activators as potential antidiabetic glucose-lowering drugs. However, the use of such substances increased the risk of hypoglycemia, and clinical studies of most synthetic GK activators are currently discontinued. Allosteric inhibitors of the GK-GKRP interaction are coming as alternative agents increasing the GK activity that can substitute GKA. In this review, we discuss the recent advances and the current state of art in the development of potential antidiabetic drugs targeted to GK as a key regulator of glucose homeostasis.

  20. Shell-type SNRs as sources of cosmic rays

    Science.gov (United States)

    Sinitsyna, V. G.; Andreeva, M. S.; Balygin, K. A.; Borisov, S. S.; Ivanov, I. A.; Kirichenko, A. M.; Klimov, A. I.; Kozhukhova, I. P.; Mirzafatikhov, R. M.; Moseiko, N. I.; Ostashev, I. E.; Palamarchuk, A. I.; Sinitsyna, V. Y.; Volokh, I. G.

    2017-06-01

    Investigations of VHE gamma-ray sources by any methods, including mirror Cherenkov telescopes, touch on the problem of the cosmic ray origin and, accordingly, the role of the Galaxy in their generation. SHALON observations have yielded results on Galactic supernova remnants (SNR) of different ages. Among them are: the shell-type SNRs Tycho's SNR (1572y), Cas A (1680y), IC 443 (age ˜ (3 ÷ 30) × 103 y), Cygni SNR (age ˜ (5 ÷ 7) × 103 y), G166.0 + 4.3 (age ˜ 24 × 103 y) and the classical nova GK Per (Nova 1901). Observation results are presented for each of the SNRs with spectral energy distributions by SHALON in comparison with other experiment data and images by SHALON together with data from X-rays by Chandra and radio-data by CGPS. The collected experimental data have confirmed the prediction of the theory about the hadronic generation mechanism of very high energy 800 GeV-100 TeV gamma-rays in Tycho's SNR, Cas A and IC443. For the first time, unique data on GK Per (Nova1901) TeV gamma-ray emission were obtained with the SHALON experiment. The X-ray data shows that the nova remnant of GK Per could be a younger remnant that will resemble older SNRs like IC 443 which interact with molecular clouds. GK Per is supposed to be a candidate for TeV gamma-ray emission due to accelerated particles in the reverse shock region.

  1. Assessment and characterization of the total geometric uncertainty in Gamma Knife radiosurgery using polymer gels

    International Nuclear Information System (INIS)

    Moutsatsos, A.; Karaiskos, P.; Pantelis, E.; Georgiou, E.; Petrokokkinos, L.; Sakelliou, L.; Torrens, M.; Seimenis, I.

    2013-01-01

    geometric uncertainty ranged from 0.34 to 0.80 mm. The average total geometric uncertainty, which also includes the GK spatial dose delivery uncertainty, was found equal to (0.88 ± 0.16), (0.88 ± 0.26), (1.02 ± 0.09), and (1.15 ± 0.24) mm for the MR image series acquired with the read gradient polarity (direction) set toward right, left, posterior, and anterior, respectively. Conclusions: The implemented methodology seems capable of assessing the total geometric uncertainty, as well as of characterizing its contributors, ascribed to the entire GK treatment delivery (i.e., from MR imaging to GK dose delivery) for an extended region of the Leksell stereotactic space. Results obtained indicate that the selection of both the frequency encoding axis and the read gradient polarity during MRI acquisition may affect the magnitude as well as the spatial components of the total geometric uncertainty.

  2. Assessment and characterization of the total geometric uncertainty in Gamma Knife radiosurgery using polymer gels.

    Science.gov (United States)

    Moutsatsos, A; Karaiskos, P; Petrokokkinos, L; Sakelliou, L; Pantelis, E; Georgiou, E; Torrens, M; Seimenis, I

    2013-03-01

    0.34 to 0.80 mm. The average total geometric uncertainty, which also includes the GK spatial dose delivery uncertainty, was found equal to (0.88 ± 0.16), (0.88 ± 0.26), (1.02 ± 0.09), and (1.15 ± 0.24) mm for the MR image series acquired with the read gradient polarity (direction) set toward right, left, posterior, and anterior, respectively. The implemented methodology seems capable of assessing the total geometric uncertainty, as well as of characterizing its contributors, ascribed to the entire GK treatment delivery (i.e., from MR imaging to GK dose delivery) for an extended region of the Leksell stereotactic space. Results obtained indicate that the selection of both the frequency encoding axis and the read gradient polarity during MRI acquisition may affect the magnitude as well as the spatial components of the total geometric uncertainty.

  3. Adaptive fractionated stereotactic Gamma Knife radiotherapy of meningioma using integrated stereotactic cone-beam-CT and adaptive re-planning (a-gkFSRT)

    International Nuclear Information System (INIS)

    Stieler, F.; Wenz, F.; Abo-Madyan, Y.; Schweizer, B.; Polednik, M.; Herskind, C.; Giordano, F.A.; Mai, S.

    2016-01-01

    The Gamma Knife Icon (Elekta AB, Stockholm, Sweden) allows frameless stereotactic treatment using a combination of cone beam computer tomography (CBCT), a thermoplastic mask system, and an infrared-based high-definition motion management (HDMM) camera system for patient tracking during treatment. We report on the first patient with meningioma at the left petrous bone treated with adaptive fractionated stereotactic radiotherapy (a-gkFSRT). The first patient treated with Gamma Knife Icon at our institute received MR imaging for preplanning before treatment. For each treatment fraction, a daily CBCT was performed to verify the actual scull/tumor position. The system automatically adapted the planned shot positions to the daily position and recalculated the dose distribution (online adaptive planning). During treatment, the HDMM system recorded the intrafractional patient motion. Furthermore, the required times were recorded to define a clinical treatment slot. Total treatment time was around 20 min. Patient positioning needed 0.8 min, CBCT positioning plus acquisition 1.65 min, CT data processing and adaptive planning 2.66 min, and treatment 15.6 min. The differences for the five daily CBCTs compared to the reference are for rotation: -0.59 ± 0.49 /0.18 ± 0.20 /0.05 ± 0.36 and for translation: 0.94 ± 0.52 mm/-0.08 ± 0.08 mm/-1.13 ± 0.89 mm. Over all fractions, an intrafractional movement of 0.13 ± 0.04 mm was observed. The Gamma Knife Icon allows combining the accuracy of the stereotactic Gamma Knife system with the flexibility of fractionated treatment with the mask system and CBCT. Furthermore, the Icon system introduces a new online patient tracking system to the clinical routine. The interfractional accuracy of patient positioning was controlled with a thermoplastic mask and CBCT. (orig.) [de

  4. THERMUS—A thermal model package for ROOT

    Science.gov (United States)

    Wheaton, S.; Cleymans, J.; Hauer, M.

    2009-01-01

    THERMUS is a package of C++ classes and functions allowing statistical-thermal model analyses of particle production in relativistic heavy-ion collisions to be performed within the ROOT framework of analysis. Calculations are possible within three statistical ensembles; a grand-canonical treatment of the conserved charges B, S and Q, a fully canonical treatment of the conserved charges, and a mixed-canonical ensemble combining a canonical treatment of strangeness with a grand-canonical treatment of baryon number and electric charge. THERMUS allows for the assignment of decay chains and detector efficiencies specific to each particle yield, which enables sensible fitting of model parameters to experimental data. Program summaryProgram title: THERMUS, version 2.1 Catalogue identifier: AEBW_v1_0 Program summary URL:http://cpc.cs.qub.ac.uk/summaries/AEBW_v1_0.html Program obtainable from: CPC Program Library, Queen's University, Belfast, N. Ireland Licensing provisions: Standard CPC licence, http://cpc.cs.qub.ac.uk/licence/licence.html No. of lines in distributed program, including test data, etc.: 17 152 No. of bytes in distributed program, including test data, etc.: 93 581 Distribution format: tar.gz Programming language: C++ Computer: PC, Pentium 4, 1 GB RAM (not hardware dependent) Operating system: Linux: FEDORA, RedHat, etc. Classification: 17.7 External routines: Numerical Recipes in C [1], ROOT [2] Nature of problem: Statistical-thermal model analyses of heavy-ion collision data require the calculation of both primordial particle densities and contributions from resonance decay. A set of thermal parameters (the number depending on the particular model imposed) and a set of thermalized particles, with their decays specified, is required as input to these models. The output is then a complete set of primordial thermal quantities for each particle, together with the contributions to the final particle yields from resonance decay. In many applications of

  5. Applicability of the PROSPECT model for Norway spruce needles

    Czech Academy of Sciences Publication Activity Database

    Malenovský, Zbyněk; Albrechtová, J.; Lhotáková, Z.; Zurita-Milla, R.; Clevers, J.G.P.W.; Schaepman, M.E.; Cudlín, Pavel

    2006-01-01

    Roč. 27, 23-24 (2006), s. 5315-5340 ISSN 0143-1161 R&D Projects: GA MŠk 1K04121 Institutional research plan: CEZ:AV0Z60870520 Keywords : chlorophyll content estimation * canopy reflectance models * radiative-transfer models * remote-sensing data * leaf-area index * optical-properties * conifer needles * forest * vegetation * inversion Subject RIV: GK - Forest ry Impact factor: 0.980, year: 2006

  6. Synthesis and biological evaluation of [11C]AZ12504948; a novel tracer for imaging of glucokinase in pancreas and liver

    International Nuclear Information System (INIS)

    Jahan, M.; Johnström, P.; Nag, S.; Takano, A.; Korsgren, O.; Johansson, L.; Halldin, C.; Eriksson, O.

    2015-01-01

    Introduction: Glucokinase (GK) is potentially a target for imaging of islets of Langerhans. Here we report the radiosynthesis and preclinical evaluation of the GK activator, [ 11 C]AZ12504948, for in vivo imaging of GK. Methods: [ 11 C]AZ12504948 was synthesized by O-methylation of the precursor, AZ125555620, using carbon-11 methyl iodide ([ 11 C]CH 3 I). Preclinical evaluation was performed by autoradiography (ARG) of human tissues and PET/CT studies in pig and non-human primate. Result: [ 11 C]AZ12504948 was produced in reproducible good radiochemical yield in 28–30 min. Radiochemical purity of the formulated product was > 98% for up to 2 h with specific radioactivities 855 ± 209 GBq/μmol (n = 8). The preclinical evaluation showed some specificity for GK in liver, but not in pancreas. Conclusion: [ 11 C]AZ12504948 images GK in liver, but the low specificity impedes the visualization of GK in pancreas. Improved target specificity is required for further progress using PET probes based on this class of GK activators

  7. Cobordism independence of Grassmann manifolds

    Indian Academy of Sciences (India)

    R. Narasimhan (Krishtel eMaging) 1461 1996 Oct 15 13:05:22

    ν(m) divides m. Given a positive integer d, let G(d) denote the set of bordism classes of all non-bounding. Grassmannian manifolds Gk(Fn+k) having real dimension d such that k < n. The restric- tion kGk(Fn+k) ≈ Gn(Fn+k) and, for k = n, Gk(Fn+k) bounds. Thus, G(d) = {[Gk(Fn+k)] ∈ 9∗ | nkt = d,k < n, ...

  8. SU-E-T-382: Evaluation of Clinical Application and Dosimetric Comparison for Treatment Plans of Gamma Knife and CyberKnife for Arteriovenous Malformations

    Energy Technology Data Exchange (ETDEWEB)

    Kuo, C [Department of Radiation Oncology, Shuang Ho Hospital, Taipei Medical Univer, New Taipei City, Taiwan (R.O.C.) (China)

    2015-06-15

    Purpose: To analyze and compare the characteristics of dose distributions between Gamma Knife (GK) and CyberKnife (CK), in treating arteriovenous malformations (AVMs), and evaluate the influences on their clinical applications. Methods: Twenty four patients with AVMs treated with CK of prescribed dose (PD) of 16–25 Gy in single fraction were selected. Each patient’s CT images used for CK treatment planning with contours of targets and critical organs were exported and then loaded into the GK planning system. GK treatment plan with the same PD used in CK was generated for each patient. The metrics for dose comparison between GK and CK included conformity index (CI), gradient index (GI) of 75%, 50% and 25% of the PD, heterogeneity index (HI), volume of brain tissues covered by 10 Gy and 12 Gy, maximum dose to brainstem and beam-on time. Paired Samples t-test was used to analyze these metrics for significance (p value). Results: The CI were 0.744 ± 0.075 (GK) and 0.768 ± 0.086 (CK), p = 0.281. The GI75%, GI50%, and GI25% in GK and CK were 1.735 ± 0.100 and 2.439 ± 0.338 (p < 0.001), 3.169 ± 0.265 and 4.972 ± 0.852 (p < 0.001), and 8.650 ± 0.914 and 14.261 ± 2.476 (p < 0.001). The HI were 0.728 ± 0.072 (GK) and 0.313 ± 0.069 (CK), p < 0.001. There were significant differences both for volume of brain tissues covered by 10 Gy and 12 Gy in GK and CK (p < 0.001). GK had smaller maximum dose to brainstem. CK had shorter beam-on time. Conclusion: GK has similar dose conformity as CK, and has better normal tissue sparing but is less efficient than CK.

  9. RT-23THE ROLE OF GAMMA KNIFE RADIOSURGERY IN THE TREATMENT FOR PRIMARY CENTRAL NERVOUS SYSTEM LYMPHOMAS

    Science.gov (United States)

    Mori, Kanji; Chiba, Yasuyoshi; Toyota, Shingo; Kumagai, Tetsuya; Yamamoto, Shota; Sugano, Hirohumi; Taki, Takuyu

    2014-01-01

    To evaluate the role of gamma knife (GK) as a treatment modality for primary central nervous system lymphomas (PCNSL), we reviewed PCNSL patients who were treated with GK between November 2004 and May 2014, retrospectively. There were 29 cases (13 males and 16 females, ranged from 33 years old to 91 years old) with 210 treated lesions. Marginal dose was from 12Gy to 18 Gy. One hundred and nineteen lesions of 23 cases could be accessed and all of them showed partial response or complete response. In seven cases, GK was performed at initial treatment, in 18 cases at recurrence, and in four cases at both. Median age of initial treatment group were significantly higher than that of recurred group (77.8 y.o. and 63.6 y.o., p = 0.00268). Two cases of initial treatment group and 16 cases of recurrent group were treated with whole brain radiation therapy (WBRT). GK after first relapse, progression free survival more than six months were obtained for six cases (33.3%), and four of them were free from additional therapy more than one year. Otherwise, thirteen cases (59.1%) received GK repeatedly (2-6 times, median = 3), and interval between each therapy was from 22 to 513 days (median = 93days). We confirmed that GK has high potential for local control. And it was supposed that GK was used as an alternative to WBRT at initial treatment for elderly, or selected to recurrent cases after WBRT. These results showed the role of GK in the treatment for PCNSL is palliative care, now in Japan. But it is clear that there exist patients who can live longer without WBRT by GK. We conclude that GK will become to play a more positive role by modifying its timing and modalities used together.

  10. SU-E-T-382: Evaluation of Clinical Application and Dosimetric Comparison for Treatment Plans of Gamma Knife and CyberKnife for Arteriovenous Malformations

    International Nuclear Information System (INIS)

    Kuo, C

    2015-01-01

    Purpose: To analyze and compare the characteristics of dose distributions between Gamma Knife (GK) and CyberKnife (CK), in treating arteriovenous malformations (AVMs), and evaluate the influences on their clinical applications. Methods: Twenty four patients with AVMs treated with CK of prescribed dose (PD) of 16–25 Gy in single fraction were selected. Each patient’s CT images used for CK treatment planning with contours of targets and critical organs were exported and then loaded into the GK planning system. GK treatment plan with the same PD used in CK was generated for each patient. The metrics for dose comparison between GK and CK included conformity index (CI), gradient index (GI) of 75%, 50% and 25% of the PD, heterogeneity index (HI), volume of brain tissues covered by 10 Gy and 12 Gy, maximum dose to brainstem and beam-on time. Paired Samples t-test was used to analyze these metrics for significance (p value). Results: The CI were 0.744 ± 0.075 (GK) and 0.768 ± 0.086 (CK), p = 0.281. The GI75%, GI50%, and GI25% in GK and CK were 1.735 ± 0.100 and 2.439 ± 0.338 (p < 0.001), 3.169 ± 0.265 and 4.972 ± 0.852 (p < 0.001), and 8.650 ± 0.914 and 14.261 ± 2.476 (p < 0.001). The HI were 0.728 ± 0.072 (GK) and 0.313 ± 0.069 (CK), p < 0.001. There were significant differences both for volume of brain tissues covered by 10 Gy and 12 Gy in GK and CK (p < 0.001). GK had smaller maximum dose to brainstem. CK had shorter beam-on time. Conclusion: GK has similar dose conformity as CK, and has better normal tissue sparing but is less efficient than CK

  11. Thermostabilization of desulfurization enzymes from Rhodococcos sp. IGTS8. Final technical report

    Energy Technology Data Exchange (ETDEWEB)

    John J. Kilbane II

    2000-12-15

    The objective of this project was to develop thermophilic cultures capable of expressing the desulfurization (dsz) operon of Rhodococcus sp. IGTS8. The approaches taken in this project included the development of plasmid and integrative expression vectors that function well in Thermus thermophilus, the cloning of Rhodococcus dsz genes in Thermus expression vectors, and the isolation of bacterial cultures that express the dsz operon at thermophilic temperatures. This project has resulted in the development of plasmid and integrative expression vectors for use in T. thermophilus. The dsz genes have been expressed at moderately thermophilic temperatures (52 C) in Mycobacterium phlei and at temperatures as high as 72 C in T. thermophilus. The tools and methods developed in this project will be generally useful for the expression of heterologous genes in Thermus. Key developments in the project have been the isolation of a Mycobacterium phlei culture capable of expressing the desulfurization operon at 52 C, development of plasmid and integrative expression vectors for Thermus thermophilus, and the development of a host-vector system based on the malate dehydrogenase gene that allows plasmids to be stably maintained in T. thermophilus and provides a convenient reporter gene for the accurate quantification of gene expression. Publications have been prepared regarding each of these topics; these preprints are included.

  12. Genetic control of differential acetylation in diabetic rats.

    Directory of Open Access Journals (Sweden)

    Pamela J Kaisaki

    Full Text Available Post-translational protein modifications such as acetylation have significant regulatory roles in metabolic processes, but their relationship to both variation in gene expression and DNA sequence is unclear. We address this question in the Goto-Kakizaki (GK rat inbred strain, a model of polygenic type 2 diabetes. Expression of the NAD-dependent deacetylase Sirtuin-3 is down-regulated in GK rats compared to normoglycemic Brown Norway (BN rats. We show first that a promoter SNP causes down-regulation of Sirtuin-3 expression in GK rats. We then use mass-spectrometry to identify proteome-wide differential lysine acetylation of putative Sirtuin-3 protein targets in livers of GK and BN rats. These include many proteins in pathways connected to diabetes and metabolic syndrome. We finally sequence GK and BN liver transcriptomes and find that mRNA expression of these targets does not differ significantly between GK and BN rats, in contrast to other components of the same pathways. We conclude that physiological differences between GK and BN rats are mediated by a combination of differential protein acetylation and gene transcription and that genetic variation can modulate acetylation independently of expression.

  13. Investigation of rheological properties of winter wheat varieties during storage

    Directory of Open Access Journals (Sweden)

    Móré M.

    2015-01-01

    Full Text Available The paper shows the results of some experimental researches on the rheological characteristics of the dough obtained from the flour of three winter wheat varieties. We used valorigraph test to determine the rheological properties of wheat flour dough, because it determines the quality of the end-products. Winter wheat varieties (Lupus, Mv Toldi and GK Csillag were produced and their samples were collected on Látókép Research Farm of the University of Debrecen in the crop year of 2011/2012. We have carried out a short-term storage experiment (from July to August, 2012. We analysed the changes in water absorption capacity, dough stability time and valorigraph quality number for 3 times (24.07.2012, 31.07.2012, 21.08.2012 during short-term storage. Our results showed that the baking quality of Lupus, Mv Toldi and GK Csillag improved during the storage period.

  14. Lys169 of human glucokinase is a determinant for glucose phosphorylation: implication for the atomic mechanism of glucokinase catalysis.

    Directory of Open Access Journals (Sweden)

    Jian Zhang

    Full Text Available Glucokinase (GK, a glucose sensor, maintains plasma glucose homeostasis via phosphorylation of glucose and is a potential therapeutic target for treating maturity-onset diabetes of the young (MODY and persistent hyperinsulinemic hypoglycemia of infancy (PHHI. To characterize the catalytic mechanism of glucose phosphorylation by GK, we combined molecular modeling, molecular dynamics (MD simulations, quantum mechanics/molecular mechanics (QM/MM calculations, experimental mutagenesis and enzymatic kinetic analysis on both wild-type and mutated GK. Our three-dimensional (3D model of the GK-Mg(2+-ATP-glucose (GMAG complex, is in agreement with a large number of mutagenesis data, and elucidates atomic information of the catalytic site in GK for glucose phosphorylation. A 10-ns MD simulation of the GMAG complex revealed that Lys169 plays a dominant role in glucose phosphorylation. This prediction was verified by experimental mutagenesis of GK (K169A and enzymatic kinetic analyses of glucose phosphorylation. QM/MM calculations were further used to study the role of Lys169 in the catalytic mechanism of the glucose phosphorylation and we found that Lys169 enhances the binding of GK with both ATP and glucose by serving as a bridge between ATP and glucose. More importantly, Lys169 directly participates in the glucose phosphorylation as a general acid catalyst. Our findings provide mechanistic details of glucose phorphorylation catalyzed by GK, and are important for understanding the pathogenic mechanism of MODY.

  15. Adaptive fractionated stereotactic Gamma Knife radiotherapy of meningioma using integrated stereotactic cone-beam-CT and adaptive re-planning (a-gkFSRT).

    Science.gov (United States)

    Stieler, F; Wenz, F; Abo-Madyan, Y; Schweizer, B; Polednik, M; Herskind, C; Giordano, F A; Mai, S

    2016-11-01

    The Gamma Knife Icon (Elekta AB, Stockholm, Sweden) allows frameless stereotactic treatment using a combination of cone beam computer tomography (CBCT), a thermoplastic mask system, and an infrared-based high-definition motion management (HDMM) camera system for patient tracking during treatment. We report on the first patient with meningioma at the left petrous bone treated with adaptive fractionated stereotactic radiotherapy (a-gkFSRT). The first patient treated with Gamma Knife Icon at our institute received MR imaging for preplanning before treatment. For each treatment fraction, a daily CBCT was performed to verify the actual scull/tumor position. The system automatically adapted the planned shot positions to the daily position and recalculated the dose distribution (online adaptive planning). During treatment, the HDMM system recorded the intrafractional patient motion. Furthermore, the required times were recorded to define a clinical treatment slot. Total treatment time was around 20 min. Patient positioning needed 0.8 min, CBCT positioning plus acquisition 1.65 min, CT data processing and adaptive planning 2.66 min, and treatment 15.6 min. The differences for the five daily CBCTs compared to the reference are for rotation: -0.59 ± 0.49°/0.18 ± 0.20°/0.05 ± 0.36° and for translation: 0.94 ± 0.52 mm/-0.08 ± 0.08 mm/-1.13 ± 0.89 mm. Over all fractions, an intrafractional movement of 0.13 ± 0.04 mm was observed. The Gamma Knife Icon allows combining the accuracy of the stereotactic Gamma Knife system with the flexibility of fractionated treatment with the mask system and CBCT. Furthermore, the Icon system introduces a new online patient tracking system to the clinical routine. The interfractional accuracy of patient positioning was controlled with a thermoplastic mask and CBCT.

  16. Guanylate kinase domains of the MAGUK family scaffold proteins as specific phospho-protein-binding modules.

    Science.gov (United States)

    Zhu, Jinwei; Shang, Yuan; Xia, Caihao; Wang, Wenning; Wen, Wenyu; Zhang, Mingjie

    2011-11-25

    Membrane-associated guanylate kinases (MAGUKs) are a large family of scaffold proteins that play essential roles in tissue developments, cell-cell communications, cell polarity control, and cellular signal transductions. Despite extensive studies over the past two decades, the functions of the signature guanylate kinase domain (GK) of MAGUKs are poorly understood. Here we show that the GK domain of DLG1/SAP97 binds to asymmetric cell division regulatory protein LGN in a phosphorylation-dependent manner. The structure of the DLG1 SH3-GK tandem in complex with a phospho-LGN peptide reveals that the GMP-binding site of GK has evolved into a specific pSer/pThr-binding pocket. Residues both N- and C-terminal to the pSer are also critical for the specific binding of the phospho-LGN peptide to GK. We further demonstrate that the previously reported GK domain-mediated interactions of DLGs with other targets, such as GKAP/DLGAP1/SAPAP1 and SPAR, are also phosphorylation dependent. Finally, we provide evidence that other MAGUK GKs also function as phospho-peptide-binding modules. The discovery of the phosphorylation-dependent MAGUK GK/target interactions indicates that MAGUK scaffold-mediated signalling complex organizations are dynamically regulated.

  17. The permeability and loss of potassium in three types of Yunnan soil measured by using 86Rb

    International Nuclear Information System (INIS)

    Liu Dayong; Bie Zhilong; Wan Zhaoliang

    1997-01-01

    The permeability and loss of potassium in three types of soil (purple, paddy and red soil) used for planting tobacco in Yunnan province were studied by using 86 Rb tracer method. The results showed that for the treatment of 0.1 gK 2 O/kg soil, the potassium loss in soils was in order of purple soil>paddy soil>red soil. The loss of potassium in purple soil and paddy soil increased with sampling time. In the same soil, the loss of potassium was fast with the treatment of 0.1 gK 2 O/kg soil, but it was slow with the treatment of 0.2 gK 2 O/kg soil and 0.3 gK 2 O/kg soil. The potassium residues in soil surface was in order of red soil>paddy soil>purple soil. And the amounts of potassium in soil surface was positively correlated with potassium added. With the increase of soil depth, a slight decrease of potassium residue was found for the treatment of 0.2 gK 2 O/kg soil and 0.3 gK 2 O/kg soil, while a slight increase for the treatment of 0.1 gK 2 O/kg soil

  18. Conserved family of glycerol kinase loci in Drosophila melanogaster

    Science.gov (United States)

    Martinez Agosto, Julian A.; McCabe, Edward R.B.

    2009-01-01

    Glycerol kinase (GK) is an enzyme that catalyzes the formation of glycerol 3-phosphate from ATP and glycerol, the rate-limiting step in glycerol utilization. We analyzed the genome of the model organism Drosophila melanogaster and identified five GK orthologs, including two loci with sequence homology to the mammalian Xp21 GK protein. Using a combination of sequence analysis and evolutionary comparisons of orthologs between species, we characterized functional domains in the protein required for GK activity. Our findings include additional conserved domains that suggest novel nuclear and mitochondrial functions for glycerol kinase in apoptosis and transcriptional regulation. Investigation of GK function in Drosophila will inform us about the role of this enzyme in development and will provide us with a tool to examine genetic modifiers of human metabolic disorders. PMID:16545593

  19. UCP2 mRNA expression is dependent on glucose metabolism in pancreatic islets

    International Nuclear Information System (INIS)

    Dalgaard, Louise T.

    2012-01-01

    Highlights: ► UCP2 mRNA levels are decreased in islets of Langerhans from glucokinase deficient mice. ► UCP2 mRNA up-regulation by glucose is dependent on glucokinase. ► Absence of UCP2 increases GSIS of glucokinase heterozygous pancreatic islets. ► This may protect glucokinase deficient mice from hyperglycemic damages. -- Abstract: Uncoupling Protein 2 (UCP2) is expressed in the pancreatic β-cell, where it partially uncouples the mitochondrial proton gradient, decreasing both ATP-production and glucose-stimulated insulin secretion (GSIS). Increased glucose levels up-regulate UCP2 mRNA and protein levels, but the mechanism for UCP2 up-regulation in response to increased glucose is unknown. The aim was to examine the effects of glucokinase (GK) deficiency on UCP2 mRNA levels and to characterize the interaction between UCP2 and GK with regard to glucose-stimulated insulin secretion in pancreatic islets. UCP2 mRNA expression was reduced in GK+/− islets and GK heterozygosity prevented glucose-induced up-regulation of islet UCP2 mRNA. In contrast to UCP2 protein function UCP2 mRNA regulation was not dependent on superoxide generation, but rather on products of glucose metabolism, because MnTBAP, a superoxide dismutase mimetic, did not prevent the glucose-induced up-regulation of UCP2. Glucose-stimulated insulin secretion was increased in UCP2−/− and GK+/− islets compared with GK+/− islets and UCP2 deficiency improved glucose tolerance of GK+/− mice. Accordingly, UCP2 deficiency increased ATP-levels of GK+/− mice. Thus, the compensatory down-regulation of UCP2 is involved in preserving the insulin secretory capacity of GK mutant mice and might also be implicated in limiting disease progression in MODY2 patients.

  20. Biochemical Changes in the Serum and Liver of albino rats exposed ...

    African Journals Online (AJOL)

    Biochemical changes in the serum and liver of albino rats chronically exposed to rats administered 5gk-1 , 7.5gk-1 and 15gk-1 of gasoline , kerosine and crude petroleum(bonny light) respectively were studied. The petroleum samples were administered intraperitoneally and the biochemical changes in the rat serum and the ...

  1. Bioprospecting of gum kondagogu (Cochlospermum gossypium) for bioremediation of uranium (VI) from aqueous solution and synthetic nuclear power reactor effluents.

    Science.gov (United States)

    Sashidhar, R B; Selvi, S Kalaignana; Vinod, V T P; Kosuri, Tanuja; Raju, D; Karuna, R

    2015-10-01

    An ecofriendly green chemistry method using a natural biopolymer, Gum Kondagogu (GK) for the removal of U (VI) from aqueous, simulated nuclear effluents was studied. The adsorption characteristic of GK towards U (VI) from aqueous solution was studied at varied pH, contact time, adsorbent dose, initial U (VI) concentration and temperature using UV-Visible spectroscopy and ICP-MS. Maximum adsorption was seen at pH 4, 0.1% GK with 60 min contact time at room temperature. The GK- U (VI) composite was characterized by FT-IR, zeta potential, TEM and SEM-EDAX. The Langmuir isotherm was found to be 487 mg of U (VI) g(-1) of GK. The adsorption capacity and (%) of U (VI) was found to be 490 ± 5.4 mg g(-1) and 98.5%. Moreover adsorption of U (VI) by GK was not influenced by other cations present in the simulated effluents. The adsorbed U (VI) was efficiently stripped from composite using 1 M HCl. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Skin-specific regulation of SREBP processing and lipid biosynthesis by glycerol kinase 5

    Science.gov (United States)

    Zhang, Duanwu; Tomisato, Wataru; Su, Lijing; Sun, Lei; Choi, Jin Huk; Zhang, Zhao; Wang, Kuan-wen; Zhan, Xiaoming; Choi, Mihwa; Li, Xiaohong; Tang, Miao; Castro-Perez, Jose M.; Hildebrand, Sara; Murray, Anne R.; Moresco, Eva Marie Y.; Beutler, Bruce

    2017-01-01

    The recessive N-ethyl-N-nitrosourea–induced phenotype toku is characterized by delayed hair growth, progressive hair loss, and excessive accumulation of dermal cholesterol, triglycerides, and ceramides. The toku phenotype was attributed to a null allele of Gk5, encoding glycerol kinase 5 (GK5), a skin-specific kinase expressed predominantly in sebaceous glands. GK5 formed a complex with the sterol regulatory element-binding proteins (SREBPs) through their C-terminal regulatory domains, inhibiting SREBP processing and activation. In Gk5toku/toku mice, transcriptionally active SREBPs accumulated in the skin, but not in the liver; they were localized to the nucleus and led to elevated lipid synthesis and subsequent hair growth defects. Similar defective hair growth was observed in kinase-inactive GK5 mutant mice. Hair growth defects of homozygous toku mice were partially rescued by treatment with the HMG-CoA reductase inhibitor simvastatin. GK5 exists as part of a skin-specific regulatory mechanism for cholesterol biosynthesis, independent of cholesterol regulation elsewhere in the body. PMID:28607088

  3. Overexpression of a novel thermostable and chloride-tolerant laccase from Thermus thermophilus SG0.5JP17-16 in Pichia pastoris and its application in synthetic dye decolorization.

    Directory of Open Access Journals (Sweden)

    Huiping Liu

    Full Text Available Laccases have been used for the decolorization and detoxification of synthetic dyes due to their ability to oxidize a wide variety of dyes with water as the sole byproduct. A putative laccase gene (LacTT from Thermus thermophilus SG0.5JP17-16 was screened using the genome mining approach, and it was highly expressed in Pichia pastoris, yielding a high laccase activity of 6130 U/L in a 10-L fermentor. The LacTT open reading frame encoded a protein of 466 amino acid residues with four putative Cu-binding regions. The optimal pH of the recombinant LacTT was 4.5, 6.0, 7.5 and 8.0 with 2,2'-azino-bis(3-ethylbenzothazoline-6-sulfonic acid (ABTS, syringaldazine (SGZ, guaiacol, and 2,6-dimethoxyphenol (2,6-DMP as the substrate, respectively. The optimal temperature of LacTT was 90°C with guaiacol as the substrate. LacTT was highly stable at pH 4.0-11.0 and thermostable at 40°C-90°C, confirming that it is a pH-stable and thermostable laccase. Furthermore, LacTT also exhibited high tolerance to halides such as NaCl, NaBr and NaF, and decolorized 100%, 94%, 94% and 73% of Congo Red, Reactive Black B and Reactive Black WNN, and Remazol Brilliant Blue R, respectively. Interestingly, addition of high concentration of NaCl increased the RBBR decolorization efficiency of LacTT. These results suggest that LacTT is a good candidate for industrial applications such as dyestuff processing and degradation of dyes in textile wastewaters.

  4. cDNA sequence and tissue expression analysis of glucokinase from ...

    African Journals Online (AJOL)

    Yomi

    2012-01-10

    Jan 10, 2012 ... distribution of GK mRNA in brain, mesenteric adipose tissue, spleen, white muscle and liver of grass ... expression profile of GK mRNA in liver normalized with β-actin level was 31, 454 and 649-fold compared .... Primers and expected products used for GK gene cDNA RT-PCR, RACE and real-time PCR.

  5. Arterial oxygen partial pressures reduce the insulin-dependent induction of the perivenously located glucokinase in rat hepatocyte cultures: mimicry of arterial oxygen pressures by H2O2.

    OpenAIRE

    Kietzmann, T; Roth, U; Freimann, S; Jungermann, K

    1997-01-01

    Liver glucokinase (GK) is localized predominantly in the perivenous zone. GK mRNA was induced by insulin maximally under venous O2 partial pressure (pO2) and only half-maximally under arterial pO2. CoCl2 and desferrioxamine mimicked venous pO2 and enhanced the insulin-dependent induction of GK mRNA under arterial pO2. H2O2 mimicked arterial pO2 and reduced insulin-induced GK mRNA under venous pO2 to the lower arterial levels. Thus the zonal O2 gradient in liver seems to have a key role in the...

  6. Dosimetric characterization of hypofractionated Gamma Knife radiosurgery of large or complex brain tumors versus linear accelerator-based treatments.

    Science.gov (United States)

    Dong, Peng; Pérez-Andújar, Angélica; Pinnaduwage, Dilini; Braunstein, Steve; Theodosopoulos, Philip; McDermott, Michael; Sneed, Penny; Ma, Lijun

    2016-12-01

    OBJECTIVE Noninvasive Gamma Knife (GK) platforms, such as the relocatable frame and on-board imaging, have enabled hypofractionated GK radiosurgery of large or complex brain lesions. This study aimed to characterize the dosimetric quality of such treatments against linear accelerator-based delivery systems that include the CyberKnife (CK) and volumetric modulated arc therapy (VMAT). METHODS Ten patients treated with VMAT at the authors' institution for large brain tumors (> 3 cm in maximum diameter) were selected for the study. The median prescription dose was 25 Gy (range 20-30 Gy) in 5 fractions. The median planning target volume (PTV) was 9.57 cm 3 (range 1.94-24.81 cm 3 ). Treatment planning was performed using Eclipse External Beam Planning V11 for VMAT on the Varian TrueBeam system, Multiplan V4.5 for the CyberKnife VSI System, and Leksell GammaPlan V10.2 for the Gamma Knife Perfexion system. The percentage of the PTV receiving at least the prescription dose was normalized to be identical across all platforms for individual cases. The prescription isodose value for the PTV, conformity index, Paddick gradient index, mean and maximum doses for organs at risk, and normal brain dose at variable isodose volumes ranging from the 5-Gy isodose volume (V5) to the 15-Gy isodose volume (V15) were compared for all of the cases. RESULTS The mean Paddick gradient index was 2.6 ± 0.2, 3.2 ± 0.5, and 4.3 ± 1.0 for GK, CK, and VMAT, respectively (p 0.06). The average prescription isodose values were 52% (range 47%-69%), 60% (range 46%-68%), and 88% (range 70%-94%) for GK, CK, and VMAT, respectively, thus producing significant variations in dose hot spots among the 3 platforms. Furthermore, the mean V5 values for GK and CK were similar (p > 0.79) at 71.9 ± 36.2 cm 3 and 73.3 ± 31.8 cm 3 , respectively, both of which were statistically lower (p linear accelerator-based treatments. Such a result supports the use of a large number of isocenters or confocal beams for the

  7. Bioprospecting of gum kondagogu (Cochlospermum gossypium) for bioremediation of uranium (VI) from aqueous solution and synthetic nuclear power reactor effluents

    International Nuclear Information System (INIS)

    Sashidhar, R.B.; Selvi, S. Kalaignana; Vinod, V.T.P.; Kosuri, Tanuja; Raju, D.; Karuna, R.

    2015-01-01

    An ecofriendly green chemistry method using a natural biopolymer, Gum Kondagogu (GK) for the removal of U (VI) from aqueous, simulated nuclear effluents was studied. The adsorption characteristic of GK towards U (VI) from aqueous solution was studied at varied pH, contact time, adsorbent dose, initial U (VI) concentration and temperature using UV–Visible spectroscopy and ICP-MS. Maximum adsorption was seen at pH 4, 0.1% GK with 60 min contact time at room temperature. The GK- U (VI) composite was characterized by FT-IR, zeta potential, TEM and SEM-EDAX. The Langmuir isotherm was found to be 487 mg of U (VI) g −1 of GK. The adsorption capacity and (%) of U (VI) was found to be 490 ± 5.4 mg g −1 and 98.5%. Moreover adsorption of U (VI) by GK was not influenced by other cations present in the simulated effluents. The adsorbed U (VI) was efficiently stripped from composite using 1 M HCl. - Highlights: • An eco-friendly method for removal of U (VI) from simulated nuclear effluents by Gum Kondagogu. • The Langmuir and Freundlich isotherm indicated favourable adsorption. • The adsorption (%) of U (VI) by GK was found to be 98.5%. • Desorption studies on biosorbed metal ions showed that HCl was a good eluent

  8. Insulin-induced activation of glycerol-3-phosphate acyltransferase by a chiro-inositol-containing insulin mediator is defective in adipocytes of insulin-resistant, type II diabetic, Goto-Kakizaki rats.

    Science.gov (United States)

    Farese, R V; Standaert, M L; Yamada, K; Huang, L C; Zhang, C; Cooper, D R; Wang, Z; Yang, Y; Suzuki, S; Toyota, T

    1994-11-08

    Type II diabetic Goto-Kakizaki (GK) rats were insulin-resistant in euglycemic-hyperinsulinemic clamp studies. We therefore examined insulin signaling systems in control Wistar and diabetic GK rats. Glycerol-3-phosphate acyltransferase (G3PAT), which is activated by headgroup mediators released from glycosyl-phosphatidylinositol (GPI), was activated by insulin in intact and cell-free adipocyte preparations of control, but not diabetic, rats. A specific chiro-inositol-containing inositol phosphoglycan (IPG) mediator, prepared from beef liver, bypassed this defect and comparably activated G3PAT in cell-free adipocyte preparations of both diabetic GK and control rats. A myo-inositol-containing IPG mediator did not activate G3PAT. Relative to control adipocytes, labeling of GPI by [3H]glucosamine was diminished by 50% and insulin failed to stimulate GPI hydrolysis in GK adipocytes. In contrast to GPI-dependent G3PAT activation, insulin-stimulated hexose transport was intact in adipocytes and soleus and gastrocnemius muscles of the GK rat, as was insulin-induced activation of mitogen-activated protein kinase and protein kinase C. We conclude that (i) chiro-inositol-containing IPG mediator activates G3PAT during insulin action, (ii) diabetic GK rats have a defect in synthesizing or releasing functional chiro-inositol-containing IPG, and (iii) defective IPG-regulated intracellular glucose metabolism contributes importantly to insulin resistance in diabetic GK rats.

  9. Critical appraisal of RapidArc radiosurgery with flattening filter free photon beams for benign brain lesions in comparison to GammaKnife: a treatment planning study.

    Science.gov (United States)

    Abacioglu, Ufuk; Ozen, Zeynep; Yilmaz, Meltem; Arifoglu, Alptekin; Gunhan, Basri; Kayalilar, Namik; Peker, Selcuk; Sengoz, Meric; Gurdalli, Salih; Cozzi, Luca

    2014-05-21

    To evaluate the role of RapidArc (RA) for stereotactic radiosurgery (SRS) of benign brain lesions in comparison to GammaKnife (GK) based technique. Twelve patients with vestibular schwannoma (VS, n = 6) or cavernous sinus meningioma (CSM, n = 6) were planned for both SRS using volumetric modulated arc therapy (VMAT) by RA. 104 MV flattening filter free photon beams with a maximum dose rate of 2400 MU/min were selected. Data were compared against plans optimised for GK. A single dose of 12.5 Gy was prescribed. The primary objective was to assess treatment plan quality. Secondary aim was to appraise treatment efficiency. For VS, comparing best GK vs. RA plans, homogeneity was 51.7 ± 3.5 vs. 6.4 ± 1.5%; Paddick conformity Index (PCI) resulted 0.81 ± 0.03 vs. 0.84 ± 0.04. Gradient index (PGI) was 2.7 ± 0.2 vs. 3.8 ± 0.6. Mean target dose was 17.1 ± 0.9 vs. 12.9 ± 0.1 Gy. For the brain stem, D(1cm3) was 5.1 ± 2.0 Gy vs 4.8 ± 1.6 Gy. For the ipsilateral cochlea, D(0.1cm3) was 1.7 ± 1.0 Gy vs. 1.8 ± 0.5 Gy. For CSM, homogeneity was 52.3 ± 2.4 vs. 12.4 ± 0.6; PCI: 0.86 ± 0.05 vs. 0.88 ± 0.05; PGI: 2.6 ± 0.1 vs. 3.8 ± 0.5; D(1cm3) to brain stem was 5.4 ± 2.8 Gy vs. 5.2 ± 2.8 Gy; D(0.1cm3) to ipsi-lateral optic nerve was 4.2 ± 2.1 vs. 2.1 ± 1.5 Gy; D(0.1cm3) to optic chiasm was 5.9 ± 3.1 vs. 4.5 ± 2.1 Gy. Treatment time was 53.7 ± 5.8 (64.9 ± 24.3) minutes for GK and 4.8 ± 1.3 (5.0 ± 0.7) minutes for RA for schwannomas (meningiomas). SRS with RA and FFF beams revealed to be adequate and comparable to GK in terms of target coverage, homogeneity, organs at risk sparing with some gain in terms of treatment efficiency.

  10. 24 CFR 84.24 - Program income.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 1 2010-04-01 2010-04-01 false Program income. 84.24 Section 84.24... income. (a) HUD shall apply the standards set forth in this section in requiring recipient organizations to account for program income related to projects financed in whole or in part with Federal funds. (b...

  11. Place of Gamma Knife Stereotactic Radiosurgery in Grade 4 Vestibular Schwannoma Based on Case Series of 86 Patients with Long-Term Follow-Up.

    Science.gov (United States)

    Lefranc, Michel; Da Roz, Leila Maria; Balossier, Anne; Thomassin, Jean Marc; Roche, Pierre Hugue; Regis, Jean

    2018-06-01

    Grade IV vestibular schwannoma (Koos classification) is generally considered to be an indication for microsurgical resection or combined radiosurgery-microsurgery. However, the place of Gamma Knife stereotactic surgery (GK-SRS), either as first-line treatment or when progression of residual tumor compresses the brainstem, has not been clearly evaluated. This article reports the results of a large case series of patients with grade 4 vestibular schwannoma treated by GK-SRS. All consecutive patients with grade IV vestibular schwannoma treated by GK-SRS in our department between 1996 and 2011 with a minimum follow-up of 3 years were included in this study. 86 patients were treated by GK-SRS with a minimum follow-up of 3 years. Mean follow-up was 6.2 years (3-16 years). The mean age of the patients at the time of GK-SRS was 54.6 years (range: 23-84) and the sex ratio was 0.6. At the time of radiosurgery, no patient presented brainstem dysfunction prior to GK-SRS. 38 patients had functional hearing before treatment. One patient presented mild trigeminal neuralgia before GK-SRS. Tumor control with no clinical deterioration was obtained in 78 patients (90.7%). No radiation-induced brainstem or cranial nerve toxicity was observed in any of these patients. Functional hearing was maintained in 25 patients. 8 (9.3%) patients presented tumor growth and required microsurgical resection in 7 cases and ventricular shunt in 1 case. On the basis of this large series, GK-SRS appears to be a safe and effective treatment option for grade IV vestibular schwannoma for patients with no signs of brainstem dysfunction. Copyright © 2018 Elsevier Inc. All rights reserved.

  12. Effects of Gingko biloba Extract on Tissue Distribution of Fluoxetine and Venlafaxine in Rats

    Directory of Open Access Journals (Sweden)

    Saad Abdulrahman Hussain

    2015-09-01

    Full Text Available Objective: There are many concerns about the interactions of herbal products with conventional drugs, which are mostly used as multiple drug treatment approach. The present study was designed to evaluate the effect of long-term use of Ginkgo biloba extract (GK on the absorption and tissue distribution of fluoxetine and venlafaxine. Materials and Methods: Forty-six Wistar rats are utilized and allocated into eight groups; 2 groups administered the vehicle and saved as control; 4 groups are treated with 100 and 200 mg/kg of GK extract for 30 days; 2 groups are treated with 40mg/kg verapamil for 10 days. The liver, kidney and brain distribution of fluoxetine and venlafaxine were evaluated after single oral doses using HPLC method. Results: 200 mg/kg GK increases fluoxetine concentrations in all studied organs, while GK 100mg/kg increases venlafaxine levels in kidney tissue and not affected in the other two organs. Conclusion: Thirty days treatment with GK (100 mg/kg increases kidney availability of venlafaxine, while 200 mg GK dose increases fluoxetine availability in the liver, kidney and brain tissues after single oral doses. [J Intercult Ethnopharmacol 2015; 4(3.000: 234-238

  13. Temporal changes in the climate sensitivity of Norway spruce andEuropean beech along an elevation gradient in Central Europe

    Czech Academy of Sciences Publication Activity Database

    Kolář, Tomáš; Čermák, P.; Trnka, Miroslav; Žid, T.; Rybníček, Michal

    2017-01-01

    Roč. 2017, č. 239 (2017), s. 24-33 ISSN 0168-1923 R&D Projects: GA MŠk(CZ) LO1415; GA ČR GA13-04291S EU Projects: European Commission(XE) EHP-CZ02-0V-1-066-01-2014 Institutional support: RVO:67179843 Keywords : czech republic * drought * fagus sylvatica * picea abies * temperature * tree-ring width chronology Subject RIV: GK - Forestry OBOR OECD: Forestry Impact factor: 3.887, year: 2016

  14. Advanced Teleprocessing Systems

    Science.gov (United States)

    1983-03-31

    Nelson and Richard Gail with equations written on napkins at lunch time. Other students of the ATS (Advanced Teleprocessing Systems) group have also been... ink Mik 2Gk (I+S,)2 Mk = (1+k/Mk) 2 - Mi k M -I- - mk -- 2Gk, Eq.(2.11) reduces to Gk - ’h (2.13) k-1 which allows throughput of G,/e for group k ( k

  15. Anti-diabetic effects of pumpkin and its components, trigonelline and nicotinic acid, on Goto-Kakizaki rats.

    Science.gov (United States)

    Yoshinari, Orie; Sato, Hideyo; Igarashi, Kiharu

    2009-05-01

    The effects of a pumpkin paste concentrate and its components on oral glucose tolerance and serum lipid levels were determined in non-obese type 2 diabetic Goto-Kakizaki (GK) rats. In the oral glucose tolerance test, the pumpkin paste concentrate-fed group maintained a lower glucose level than the control group between 15 and 60 min. The compounds considered to be effective in improving glucose tolerance and contained in the methanol extract of the pumpkin in relatively abundant amounts were isolated and identified as trigonelline (TRG) and nicotinic acid (NA).Feeding a diet containing TRG and NA respectively improved and tended to improve glucose tolerance. The insulin level increased after 15 min in the TRG-fed GK rats and then gradually decreased over the next 120 min. In contrast, a gradual increase was seen in the insulin level over 120 min in the control GK rats not fed with TRG, suggesting that TRG could improve the insulin resistance. The serum and liver triglyceride (TG) levels in the TRG- and NA-fed GK rats were lower than those in the control GK rats. Lower activity of liver fatty acid synthase (FAS), and higher activity of liver carnitine palmitoyl transferase (CPT) and glucokinase (GLK) in the TRG- and NA-fed GK rats than in the control GK rats were observed. This suggests that the regulation of these enzyme activities by TRG and NA was closely related to the suppression of both TG accumulation and the progression of diabetes.

  16. Comparison of INTAKE24 (an Online 24-h Dietary Recall Tool) with Interviewer-Led 24-h Recall in 11-24 Year-Old.

    Science.gov (United States)

    Bradley, Jennifer; Simpson, Emma; Poliakov, Ivan; Matthews, John N S; Olivier, Patrick; Adamson, Ashley J; Foster, Emma

    2016-06-09

    Online dietary assessment tools offer a convenient, low cost alternative to traditional dietary assessment methods such as weighed records and face-to-face interviewer-led 24-h recalls. INTAKE24 is an online multiple pass 24-h recall tool developed for use with 11-24 year-old. The aim of the study was to undertake a comparison of INTAKE24 (the test method) with interviewer-led multiple pass 24-h recalls (the comparison method) in 180 people aged 11-24 years. Each participant completed both an INTAKE24 24-h recall and an interviewer-led 24-h recall on the same day on four occasions over a one-month period. The daily energy and nutrient intakes reported in INTAKE24 were compared to those reported in the interviewer-led recall. Mean intakes reported using INTAKE24 were similar to the intakes reported in the interviewer-led recall for energy and macronutrients. INTAKE24 was found to underestimate energy intake by 1% on average compared to the interviewer-led recall with the limits of agreement ranging from minus 49% to plus 93%. Mean intakes of all macronutrients and micronutrients (except non-milk extrinsic sugars) were within 4% of the interviewer-led recall. Dietary assessment that utilises technology may offer a viable alternative and be more engaging than paper based methods, particularly for children and young adults.

  17. EST Table: FS737373 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FS737373 E_FL_bmmt_08K06_R_0 10/09/28 37 %/161 aa ref|XP_002067709.1| GK12566 [Dros...ophila willistoni] gb|EDW78695.1| GK12566 [Drosophila willistoni] 10/09/03 37 %/161 aa FBpp0241709|DwilGK12566-PA 10/08/28 n.h 10/09/10 n.h 10/09/10 n.h 10/09/10 n.h FS737272 bmmt ...

  18. Genomic-Enabled Prediction in Maize Using Kernel Models with Genotype × Environment Interaction.

    Science.gov (United States)

    Bandeira E Sousa, Massaine; Cuevas, Jaime; de Oliveira Couto, Evellyn Giselly; Pérez-Rodríguez, Paulino; Jarquín, Diego; Fritsche-Neto, Roberto; Burgueño, Juan; Crossa, Jose

    2017-06-07

    Multi-environment trials are routinely conducted in plant breeding to select candidates for the next selection cycle. In this study, we compare the prediction accuracy of four developed genomic-enabled prediction models: (1) single-environment, main genotypic effect model (SM); (2) multi-environment, main genotypic effects model (MM); (3) multi-environment, single variance G×E deviation model (MDs); and (4) multi-environment, environment-specific variance G×E deviation model (MDe). Each of these four models were fitted using two kernel methods: a linear kernel Genomic Best Linear Unbiased Predictor, GBLUP (GB), and a nonlinear kernel Gaussian kernel (GK). The eight model-method combinations were applied to two extensive Brazilian maize data sets (HEL and USP data sets), having different numbers of maize hybrids evaluated in different environments for grain yield (GY), plant height (PH), and ear height (EH). Results show that the MDe and the MDs models fitted with the Gaussian kernel (MDe-GK, and MDs-GK) had the highest prediction accuracy. For GY in the HEL data set, the increase in prediction accuracy of SM-GK over SM-GB ranged from 9 to 32%. For the MM, MDs, and MDe models, the increase in prediction accuracy of GK over GB ranged from 9 to 49%. For GY in the USP data set, the increase in prediction accuracy of SM-GK over SM-GB ranged from 0 to 7%. For the MM, MDs, and MDe models, the increase in prediction accuracy of GK over GB ranged from 34 to 70%. For traits PH and EH, gains in prediction accuracy of models with GK compared to models with GB were smaller than those achieved in GY. Also, these gains in prediction accuracy decreased when a more difficult prediction problem was studied. Copyright © 2017 Bandeira e Sousa et al.

  19. Genomic-Enabled Prediction in Maize Using Kernel Models with Genotype × Environment Interaction

    Directory of Open Access Journals (Sweden)

    Massaine Bandeira e Sousa

    2017-06-01

    Full Text Available Multi-environment trials are routinely conducted in plant breeding to select candidates for the next selection cycle. In this study, we compare the prediction accuracy of four developed genomic-enabled prediction models: (1 single-environment, main genotypic effect model (SM; (2 multi-environment, main genotypic effects model (MM; (3 multi-environment, single variance G×E deviation model (MDs; and (4 multi-environment, environment-specific variance G×E deviation model (MDe. Each of these four models were fitted using two kernel methods: a linear kernel Genomic Best Linear Unbiased Predictor, GBLUP (GB, and a nonlinear kernel Gaussian kernel (GK. The eight model-method combinations were applied to two extensive Brazilian maize data sets (HEL and USP data sets, having different numbers of maize hybrids evaluated in different environments for grain yield (GY, plant height (PH, and ear height (EH. Results show that the MDe and the MDs models fitted with the Gaussian kernel (MDe-GK, and MDs-GK had the highest prediction accuracy. For GY in the HEL data set, the increase in prediction accuracy of SM-GK over SM-GB ranged from 9 to 32%. For the MM, MDs, and MDe models, the increase in prediction accuracy of GK over GB ranged from 9 to 49%. For GY in the USP data set, the increase in prediction accuracy of SM-GK over SM-GB ranged from 0 to 7%. For the MM, MDs, and MDe models, the increase in prediction accuracy of GK over GB ranged from 34 to 70%. For traits PH and EH, gains in prediction accuracy of models with GK compared to models with GB were smaller than those achieved in GY. Also, these gains in prediction accuracy decreased when a more difficult prediction problem was studied.

  20. Comparison of INTAKE24 (an Online 24-h Dietary Recall Tool with Interviewer-Led 24-h Recall in 11–24 Year-Old

    Directory of Open Access Journals (Sweden)

    Jennifer Bradley

    2016-06-01

    Full Text Available Online dietary assessment tools offer a convenient, low cost alternative to traditional dietary assessment methods such as weighed records and face-to-face interviewer-led 24-h recalls. INTAKE24 is an online multiple pass 24-h recall tool developed for use with 11–24 year-old. The aim of the study was to undertake a comparison of INTAKE24 (the test method with interviewer-led multiple pass 24-h recalls (the comparison method in 180 people aged 11–24 years. Each participant completed both an INTAKE24 24-h recall and an interviewer-led 24-h recall on the same day on four occasions over a one-month period. The daily energy and nutrient intakes reported in INTAKE24 were compared to those reported in the interviewer-led recall. Mean intakes reported using INTAKE24 were similar to the intakes reported in the interviewer-led recall for energy and macronutrients. INTAKE24 was found to underestimate energy intake by 1% on average compared to the interviewer-led recall with the limits of agreement ranging from minus 49% to plus 93%. Mean intakes of all macronutrients and micronutrients (except non-milk extrinsic sugars were within 4% of the interviewer-led recall. Dietary assessment that utilises technology may offer a viable alternative and be more engaging than paper based methods, particularly for children and young adults.

  1. ImmunoPET Imaging of Murine CD4+ T Cells Using Anti-CD4 Cys-Diabody: Effects of Protein Dose on T Cell Function and Imaging.

    Science.gov (United States)

    Freise, Amanda C; Zettlitz, Kirstin A; Salazar, Felix B; Lu, Xiang; Tavaré, Richard; Wu, Anna M

    2017-08-01

    Molecular imaging of CD4 + T cells throughout the body has implications for monitoring autoimmune disease and immunotherapy of cancer. Given the key role of these cells in regulating immunity, it is important to develop a biologically inert probe. GK1.5 cys-diabody (cDb), a previously developed anti-mouse CD4 antibody fragment, was tested at different doses to assess its effects on positron emission tomography (PET) imaging and CD4 + T cell viability, proliferation, CD4 expression, and function. The effect of protein dose on image contrast (lymphoid tissue-to-muscle ratio) was assessed by administering different amounts of 89 Zr-labeled GK1.5 cDb to mice followed by PET imaging and ex vivo biodistribution analysis. To assess impact of GK1.5 cDb on T cell biology, GK1.5 cDb was incubated with T cells in vitro or administered intravenously to C57BL/6 mice at multiple protein doses. CD4 expression and T cell proliferation were analyzed with flow cytometry and cytokines were assayed. For immunoPET imaging, the lowest protein dose of 2 μg of 89 Zr-labeled GK1.5 cDb resulted in significantly higher % injected dose/g in inguinal lymph nodes (ILN) and spleen compared to the 12-μg protein dose. In vivo administration of GK1.5 cDb at the high dose of 40 μg caused a transient decrease in CD4 expression in spleen, blood, lymph nodes, and thymus, which recovered within 3 days postinjection; this effect was reduced, although not abrogated, when 2 μg was administered. Proliferation was inhibited in vivo in ILN but not the spleen by injection of 40 μg GK1.5 cDb. Concentrations of GK1.5 cDb in excess of 25 nM significantly inhibited CD4 + T cell proliferation and interferon-γ production in vitro. Overall, using low-dose GK1.5 cDb minimized biological effects on CD4 + T cells. Low-dose GK1.5 cDb yields high-contrast immunoPET images with minimal effects on T cell biology in vitro and in vivo and may be a useful tool for investigating CD4 + T cells in the context of

  2. Mercury and methylmercury intake estimation due to seafood products for the Catalonian population (Spain)

    OpenAIRE

    2009-01-01

    Abstract This study estimates mercury and methylmercury levels in fish and fishery products commercialized in the city of Barcelona from 2001 to 2007. Combining food levels data with the consumption data of 2158 people (as the median of two 24-hour-recall), the total Mercury intake of the Catalonian population was calculated. Mercury was detected in 32,8% of analyses samples. The general population average weekly intake of total mercury in Catalonian population was 0.783 ?g/k...

  3. Molecular docking and molecular dynamics simulation studies on Thermus thermophilus leucyl-tRNA synthetase complexed with different amino acids and pre-transfer editing substrates

    Directory of Open Access Journals (Sweden)

    Rayevsky A. V.

    2016-02-01

    Full Text Available Aim. To investigate the structural bases for the amino acid selectivity of the Thermus thermophilus leucyl-tRNA synthetase (LeuRSTT aminoacylation site and to disclose the binding pattern of pre-transfer editing substrates. Methods. Eight amino acids proposed as semi-cognate substrates for aminoacylation and eight aminoacyl-adenylates (formed from AMP and eight amino acids were prepared in zwitterions form. The protein structure with a co-crystallized substrate in the aminoacylation site [PDBID: 1OBH] was taken from RCSB. Docking settings and evaluation of substrate efficiency were followed by twofold docking function analysis for each conformation with Gold CCDC. The molecular dynamics simulation was performed using Gromacs. The procedures of relaxation and binding study were separated in two different subsequent simulations for 50ns and 5ns. Results. The evaluation of substrate efficiency for 8 amino acids by twofold docking function analysis, based on score values,has shown that the ligands of LeuRSTT can be positioned in the following order: Leu>Nva>Hcy>Nle>Met>Cys>Ile >Val. MD simulation has revealed lower electrostatic interactions of isoleucine with the active site of the enzyme compared with those for norvaline and leucine. In the case of aminoacyl-adenylates no significant differences were found based on score values for both GoldScore and Asp functions. Molecular dynamics of leucyl-, isoleucyl- and norvalyl-adenylates showed that the most stable and conformationally favorable is leucine, then follow norvaline and isoleucine. It has been also found that the TYR43 of the active site covers carboxyl group of leucine and norvaline like a shield and deflected towards isoleucine, allowing water molecules to come closer. Conclusions. In this study we revealed some structural basis for screening unfavorable substrates by shape, size and flexibility of a radical. The results obtained for different amino acids by molecular docking and MD studies

  4. Optimal low-dose radiosurgery combined with and without intervention for unruptured arteriovenous malformations. A long-term follow-up study

    International Nuclear Information System (INIS)

    Inoue, Hiroshi K.; Naitou, Isao

    2006-01-01

    Treatment indication for unruptured arteriovenous malformations (AVMs) of the brain was investigated based on long-term results of low-dose Gamma Knife (GK) surgery. Thirty-nine patients with unruptured AVMs treated with GK surgery were followed more than 7 years. These AVMs were found due to seizure and steal symptoms in 22 and 4 patients, respectively, and incidentally in 13. Embolization was performed in 6 patients before GK surgery. Total obliteration was obtained in 83.3% after initial treatment and 95.8% after additional treatment. Bleeding before total obliteration occurred in 2 patients and adverse effect on the internal capsule in 1. It is concluded that low-dose GK surgery is safe and effective for the treatment of unruptured AVMs, especially symptomatic AVMs in functional areas. (author)

  5. 4 CFR 22.24 - Alternative Dispute Resolution [Rule 24].

    Science.gov (United States)

    2010-01-01

    ... 4 Accounts 1 2010-01-01 2010-01-01 false Alternative Dispute Resolution [Rule 24]. 22.24 Section... GOVERNMENT ACCOUNTABILITY OFFICE CONTRACT APPEALS BOARD § 22.24 Alternative Dispute Resolution [Rule 24]. (a) Docketed appeals. The Board considers Alternative Dispute Resolution (ADR) to be an efficient way to timely...

  6. Positional differences in the cardiorespiratory, autonomic, and somatic profiles of professional soccer players

    Directory of Open Access Journals (Sweden)

    Martin Semjon

    2016-06-01

    Full Text Available Background: In order to optimize training in soccer, knowledge about the specific position fitness demands and characteristics is required. Objective: The aim of the study was to determine whether there are position specific differences in somatic, cardiorespiratory and autonomic cardiac profiles of professional senior Czech soccer players. Methods: All players (N = 120 were divided into six groups according to field positions: goalkeeper (GK; n = 11, external defenders (ED; n = 15, central defenders (CD; n = 18, external midfielders (EM; n = 18, central midfielders (CM; n = 24 and forwards (F; n = 34. Players underwent anthropometrical and heart rate variability (HRV assessment, and a maximal incremental running test in order to obtain maximal oxygen uptake (VO2max and heart rate. HRV variables were transformed using the natural logarithm (Ln. Results: GK and CD were significantly (p = .005 heavier than ED, EM, CM, and F; while F were significantly (p = .026 heavier than CM and EM. GK and CD were significantly (p = .008 taller than ED, EM, CM, F; and EM were significantly (p = .041 shorter than CM and F. The only significant (p = .043 difference in percentage of body fat was observed in CD compared with CM. A significantly (p = .021 lower VO2max was observed for GK compared with ED, CD, EM, CM, and F. Supine HRV was significantly (p = .039 lower for Ln LF/HF in GK compared with F. Standing HRV was significantly (p = .03 lower for Ln LF in CD compared with both ED and F, significantly (p = .028 higher for Ln LF/HF in ED compared with CD and EM. Conclusions: In soccer, specific positions are associated with different height, body mass and aerobic capacity. A lower VO2max and vagal activity in GK compared with other playing positions may not be considered as disadvantage for performance in this specific playing position.

  7. Results of Gamma Knife Radiosurgery in Acromegaly

    Directory of Open Access Journals (Sweden)

    Alberto Franzin

    2012-01-01

    Full Text Available Objective. Single-session radiosurgery with Gamma Knife (GK may be a potential adjuvant treatment in acromegaly. We analyzed the safety and efficacy of GK in patients who had previously received maximal surgical debulking at our hospital. Methods. The study was a retrospective analysis of hormonal, radiological, and ophthalmologic data collected in a predefined protocol from 1994 to 2009. The mean age at treatment was 42.3 years (range 22–67 yy. 103 acromegalic patients participated in the study. The median follow-up was 71 months (IQ range 43–107. All patients were treated with GK for residual or recurrent GH-secreting adenoma. Results. Sixty-three patients (61.2% reached the main outcome of the study. The rate of remission was 58.3% at 5 years (95% CI 47.6–69.0%. Other 15 patients (14.6% were in remission after GK while on treatment with somatostatin analogues. No serious side effects occurred after GK. Eight patients (7.8% experienced a new deficit of pituitary function. New cases of hypogonadism, hypothyroidism, and hypoadrenalism occurred in 4 of 77 patients (5.2%, 3 of 95 patients (3.2%, and 6 of 100 patients at risk (6.0%, respectively. Conclusion. In a highly selected group of acromegalic patients, GK treatment had good efficacy and safety.

  8. Relación entre conductancias de sodio y potasio al estilo Hodgkin y Huxley y su efecto en la propagación de potenciales de acción a 40°C

    Directory of Open Access Journals (Sweden)

    Oscar Emilio Hernández Bustos

    2011-01-01

    Full Text Available Objetivo: Determinar, a partir de simulacion computacional, la respuesta de un axon no mielinado a un estimulo de corriente, para diferentes valores de GNa y GK tales que 0S/ cm2GK.2S/cm2 y a 40°C Metodologia: Se simulo con el software NEURON la propagacion de potenciales de accion a traves de un modelo neuronal llamado neurox. A este se le insertaron propiedades pasivas y canales de sodio y potasio al estilo HH, cuyas tasas de apertura y cierre fueron corregidas con el coeficiente termico ¿È. Resultados: Existe una region de puntos en el espacio GNa vs GK, limitada por las rectas GNa = 12GK + 1.4 y GNa = 2.9 GK + 0.73, y , en la cual se propaga un potencial de accion a traves de neurox a 40°C. Conclusion: Las conductancias de sodio y potasio al estilo HH pueden existir en neuronas que funcionan a 40 °C con multiples posibilidades de combinaciones (GNa, GK que permiten superar las limitaciones impuestas por el aumento en la velocidad de apertura y cierre de los canales correspondientes. Este trabajo establecio un referente metodologico para la exploracion de otros espacios de combinaciones de conductancias.

  9. Changing practice patterns of Gamma Knife versus linear accelerator-based stereotactic radiosurgery for brain metastases in the US.

    Science.gov (United States)

    Park, Henry S; Wang, Elyn H; Rutter, Charles E; Corso, Christopher D; Chiang, Veronica L; Yu, James B

    2016-04-01

    Single-fraction stereotactic radiosurgery (SRS) is a crucial component in the management of limited brain metastases from non-small cell lung cancer (NSCLC). Intracranial SRS has traditionally been delivered using a frame-based Gamma Knife (GK) platform, but stereotactic modifications to the linear accelerator (LINAC) have made an alternative approach possible. In the absence of definitive prospective trials comparing the efficacy and toxicities of treatment between the 2 techniques, nonclinical factors (such as technology accessibility, costs, and efficiency) may play a larger role in determining which radiosurgery system a facility may choose to install. To the authors' knowledge, this study is the first to investigate national patterns of GK SRS versus LINAC SRS use and to determine which factors may be associated with the adoption of these radiosurgery systems. The National Cancer Data Base was used to identify patients > 18 years old with NSCLC who were treated with single-fraction SRS to the brain between 2003 and 2011. Patients who received "SRS not otherwise specified" or who did not receive a radiotherapy dose within the range of 12-24 Gy were excluded to reduce the potential for misclassification. The chi-square test, t-test, and multivariable logistic regression analysis were used to compare potential demographic, clinicopathologic, and health care system predictors of GK versus LINAC SRS use, when appropriate. This study included 1780 patients, among whom 1371 (77.0%) received GK SRS and 409 (23.0%) underwent LINAC SRS. Over time, the proportion of patients undergoing LINAC SRS steadily increased, from 3.2% in 2003 to 30.8% in 2011 (p < 0.001). LINAC SRS was adopted more rapidly by community versus academic facilities (overall 29.2% vs 17.2%, p < 0.001). On multivariable analysis, 4 independent predictors of increased LINAC SRS use emerged, including year of diagnosis in 2008-2011 versus 2003-2007 (adjusted OR [AOR] 2.04, 95% CI 1.52-2.73, p < 0

  10. Current transport and mass separation for an asymmetric fluctuation system with correlated noises

    Science.gov (United States)

    Wang, Jie; Ning, Li-Juan

    2018-01-01

    Not Available Projected supported by the National Natural Science Foundation of China (Grant No. 11202120) and the Fundamental Research Funds for the Central Universities of China (Grant Nos. GK201502007 and GK201701001).

  11. Bleeding Efficiency and Meat Oxidative Stability and Microbiological Quality of New Zealand White Rabbits Subjected to Halal Slaughter without Stunning and Gas Stun-killing

    Science.gov (United States)

    Nakyinsige, K.; Fatimah, A. B.; Aghwan, Z. A.; Zulkifli, I.; Goh, Y. M.; Sazili, A. Q.

    2014-01-01

    A study was conducted to compare the effect of halal slaughter without stunning and gas stun killing followed by bleeding on residual blood content and storage stability of rabbit meat. Eighty male New Zealand white rabbits were divided into two groups of 40 animals each and subjected to either halal slaughter without stunning (HS) or gas stun-kill (GK). The volume of blood lost during exsanguination was measured. Residual blood was further quantified by determination of haemoglobin content in Longissimus lumborum (LL) muscle. Storage stability of the meat was evaluated by microbiological analysis and measuring lipid oxidation in terms of thiobarbituric acid reactive substances (TBARS). HS resulted in significantly higher blood loss than GK. HS had significantly lower residual haemoglobin in LL muscle compared to GK. Slaughter method had no effect on rabbit meat lipid oxidation at 0, 1, and 3 d postmortem. However, at 5 and 8 days of storage at 4°C, significant differences (p<0.05) were found, with meat from the GK group exhibiting significantly higher levels of MDA than that from HS. At day 3, greater growth of Pseudomonas aeroginosa and E. coli were observed in the GK group (p<0.05) with B. thermosphacta and total aerobic counts remained unaffected by slaughter method. At days 5 and 7 postmortem, bacterial counts for all tested microbes were affected by slaughter method, with GK exhibiting significantly higher growth than HS. It can be concluded that slaughter method can affect keeping quality of rabbit meat, and HS may be a favourable option compared to GK due to high bleed out. PMID:25049968

  12. FoxO1 and HNF-4 are involved in regulation of hepatic glucokinase gene expression by resveratrol.

    Science.gov (United States)

    Ganjam, Goutham Kumar; Dimova, Elitsa Y; Unterman, Terry G; Kietzmann, Thomas

    2009-11-06

    Resveratrol, a polyphenol derived from grapes, exerts important effects on glucose and lipid metabolism, yet detailed mechanisms mediating these effects remain unknown. The liver plays a central role in energy homeostasis, and glucokinase (GK) is a key enzyme involved in glucose utilization. Resveratrol activates SIRT1 (sirtuin 1), which promotes deacetylation of the forkhead transcription factor FoxO1. Previously, we reported that FoxO1 can suppress and that HNF-4 can stimulate GK expression in the liver. Here, we examined the role of FoxO1 and HNF-4 in mediating resveratrol effects on liver GK expression. Resveratrol suppressed hepatic GK expression in vivo and in isolated hepatocytes, and knocking down FoxO1 with shRNAs disrupted this effect. Reporter gene, gel shift, supershift assay, and chromatin immunoprecipitation studies show that FoxO1 binds to the GK promoter and that the interplay between FoxO1 and HNF-4 within the GK promoter is essential for mediating the effects of resveratrol. Resveratrol promotes deacetylation of FoxO1 and enhances its recruitment to the FoxO-binding element. Conversely, resveratrol suppresses recruitment of HNF-4 to its binding site, and knockdown of FoxO1 blocks this effect of resveratrol. Coprecipitation and chromatin immunoprecipitation studies show that resveratrol enhances interaction between FoxO1 and HNF-4, reduces binding of HNF-4 to its own site, and promotes its recruitment to the FoxO site in a FoxO1-dependent manner. These results provide the first evidence that resveratrol represses GK expression via FoxO1 and that the interaction between FoxO1 and HNF-4 contributes to these effects of resveratrol.

  13. Bleeding Efficiency and Meat Oxidative Stability and Microbiological Quality of New Zealand White Rabbits Subjected to Halal Slaughter without Stunning and Gas Stun-killing

    Directory of Open Access Journals (Sweden)

    K. Nakyinsige

    2014-03-01

    Full Text Available A study was conducted to compare the effect of halal slaughter without stunning and gas stun killing followed by bleeding on residual blood content and storage stability of rabbit meat. Eighty male New Zealand white rabbits were divided into two groups of 40 animals each and subjected to either halal slaughter without stunning (HS or gas stun-kill (GK. The volume of blood lost during exsanguination was measured. Residual blood was further quantified by determination of haemoglobin content in Longissimus lumborum (LL muscle. Storage stability of the meat was evaluated by microbiological analysis and measuring lipid oxidation in terms of thiobarbituric acid reactive substances (TBARS. HS resulted in significantly higher blood loss than GK. HS had significantly lower residual haemoglobin in LL muscle compared to GK. Slaughter method had no effect on rabbit meat lipid oxidation at 0, 1, and 3 d postmortem. However, at 5 and 8 days of storage at 4°C, significant differences (p<0.05 were found, with meat from the GK group exhibiting significantly higher levels of MDA than that from HS. At day 3, greater growth of Pseudomonas aeroginosa and E. coli were observed in the GK group (p<0.05 with B. thermosphacta and total aerobic counts remained unaffected by slaughter method. At days 5 and 7 postmortem, bacterial counts for all tested microbes were affected by slaughter method, with GK exhibiting significantly higher growth than HS. It can be concluded that slaughter method can affect keeping quality of rabbit meat, and HS may be a favourable option compared to GK due to high bleed out.

  14. Bleeding Efficiency and Meat Oxidative Stability and Microbiological Quality of New Zealand White Rabbits Subjected to Halal Slaughter without Stunning and Gas Stun-killing.

    Science.gov (United States)

    Nakyinsige, K; Fatimah, A B; Aghwan, Z A; Zulkifli, I; Goh, Y M; Sazili, A Q

    2014-03-01

    A study was conducted to compare the effect of halal slaughter without stunning and gas stun killing followed by bleeding on residual blood content and storage stability of rabbit meat. Eighty male New Zealand white rabbits were divided into two groups of 40 animals each and subjected to either halal slaughter without stunning (HS) or gas stun-kill (GK). The volume of blood lost during exsanguination was measured. Residual blood was further quantified by determination of haemoglobin content in Longissimus lumborum (LL) muscle. Storage stability of the meat was evaluated by microbiological analysis and measuring lipid oxidation in terms of thiobarbituric acid reactive substances (TBARS). HS resulted in significantly higher blood loss than GK. HS had significantly lower residual haemoglobin in LL muscle compared to GK. Slaughter method had no effect on rabbit meat lipid oxidation at 0, 1, and 3 d postmortem. However, at 5 and 8 days of storage at 4°C, significant differences (p<0.05) were found, with meat from the GK group exhibiting significantly higher levels of MDA than that from HS. At day 3, greater growth of Pseudomonas aeroginosa and E. coli were observed in the GK group (p<0.05) with B. thermosphacta and total aerobic counts remained unaffected by slaughter method. At days 5 and 7 postmortem, bacterial counts for all tested microbes were affected by slaughter method, with GK exhibiting significantly higher growth than HS. It can be concluded that slaughter method can affect keeping quality of rabbit meat, and HS may be a favourable option compared to GK due to high bleed out.

  15. EST Table: FY036562 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available FY036562 rbmte12o02 11/11/04 33 %/193 aa ref|XP_002070433.1| GK11033 [Drosophila willistoni] gb|EDW81419.1| GK11033 [Drosophila willistoni] 11/11/04 32 %/183 aa FBpp0240176|DwilGK11033-PA 11/11/04 n.h 11/11/04 low homology 11/11/04 n.h 11/11/04 30 %/181 aa gi|189238448|ref|XP_001813701.1| PREDICTED: similar to Fanconi anemia complementation group L [Tribolium castaneum] FS903257 bmte ...

  16. Prevention and reversal of experimental autoimmune thyroiditis (EAT) in mice by administration of anti-L3T4 monoclonal antibody at different stages of disease development.

    Science.gov (United States)

    Stull, S J; Kyriakos, M; Sharp, G C; Braley-Mullen, H

    1988-11-01

    Experimental autoimmune thyroiditis (EAT) can be induced in CBA/J mice following the transfer of spleen cells from mouse thyroglobulin (MTg)-sensitized donors that have been activated in vitro with MTg. Since L3T4+ T cells are required to transfer EAT in this model, the present study was undertaken to assess the effectiveness of the anti-L3T4 monoclonal antibody (mAb) GK1.5 in preventing or arresting the development of EAT. Spleen cells from mice given mAb GK1.5 prior to sensitization with MTg and adjuvant could not transfer EAT to normal recipients and cells from these mice did not proliferate in vitro to MTg. Donor mice given GK1.5 before immunization did not develop anti-MTg autoantibody and recipients of cells from such mice also produced little anti-MTg. GK1.5 could also prevent the proliferation and activation of sensitized effector cell precursors when added to in vitro cultures. When a single injection of mAb GK1.5 was given to recipients of in vitro-activated spleen cells, EAT was reduced whether the mAb was given prior to cell transfer or as late as 19 days after cell transfer. Whereas the incidence and severity of EAT was consistently reduced by injecting recipient mice with GK1.5, the same mice generally had no reduction in anti-MTg autoantibody. Since EAT is consistently induced in control recipients by 14-19 days after cell transfer, the ability of mAb GK1.5 to inhibit EAT when injected 14 or 19 days after cell transfer indicates that a single injection of the mAb GK1.5 can cause reversal of the histopathologic lesions of EAT in mice. These studies further establish the important role of L3T4+ T cells in the pathogenesis of EAT in mice and also suggest that therapy with an appropriate mAb may be an effective treatment for certain autoimmune diseases even when the therapy is initiated late in the course of the disease.

  17. Functional Characterization of MODY2 Mutations Highlights the Importance of the Fine-Tuning of Glucokinase and Its Role in Glucose Sensing

    Science.gov (United States)

    García-Herrero, Carmen-María; Rubio-Cabezas, Oscar; Azriel, Sharona; Gutierrez-Nogués, Angel; Aragonés, Angel; Vincent, Olivier; Campos-Barros, Angel; Argente, Jesús; Navas, María-Angeles

    2012-01-01

    Glucokinase (GK) acts as a glucose sensor in the pancreatic beta-cell and regulates insulin secretion. Heterozygous mutations in the human GK-encoding GCK gene that reduce the activity index increase the glucose-stimulated insulin secretion threshold and cause familial, mild fasting hyperglycaemia, also known as Maturity Onset Diabetes of the Young type 2 (MODY2). Here we describe the biochemical characterization of five missense GK mutations: p.Ile130Thr, p.Asp205His, p.Gly223Ser, p.His416Arg and p.Ala449Thr. The enzymatic analysis of the corresponding bacterially expressed GST-GK mutant proteins show that all of them impair the kinetic characteristics of the enzyme. In keeping with their position within the protein, mutations p.Ile130Thr, p.Asp205His, p.Gly223Ser, and p.His416Arg strongly decrease the activity index of GK, affecting to one or more kinetic parameters. In contrast, the p.Ala449Thr mutation, which is located in the allosteric activator site, does not affect significantly the activity index of GK, but dramatically modifies the main kinetic parameters responsible for the function of this enzyme as a glucose sensor. The reduced Kcat of the mutant (3.21±0.28 s−1 vs 47.86±2.78 s−1) is balanced by an increased glucose affinity (S0.5 = 1.33±0.08 mM vs 7.86±0.09 mM) and loss of cooperativity for this substrate. We further studied the mechanism by which this mutation impaired GK kinetics by measuring the differential effects of several competitive inhibitors and one allosteric activator on the mutant protein. Our results suggest that this mutation alters the equilibrium between the conformational states of glucokinase and highlights the importance of the fine-tuning of GK and its role in glucose sensing. PMID:22291974

  18. Nonequilibrium Gyrokinetic Fluctuation Theory and Sampling Noise in Gyrokinetic Particle-in-cell Simulations

    International Nuclear Information System (INIS)

    Krommes, John A.

    2007-01-01

    The present state of the theory of fluctuations in gyrokinetic (GK) plasmas and especially its application to sampling noise in GK particle-in-cell (PIC) simulations is reviewed. Topics addressed include the Δf method, the fluctuation-dissipation theorem for both classical and GK many-body plasmas, the Klimontovich formalism, sampling noise in PIC simulations, statistical closure for partial differential equations, the theoretical foundations of spectral balance in the presence of arbitrary noise sources, and the derivation of Kadomtsev-type equations from the general formalism

  19. Nonequilibrium Gyrokinetic Fluctuation Theory and Sampling Noise in Gyrokinetic Particle-in-cell Simulations

    Energy Technology Data Exchange (ETDEWEB)

    John A. Krommes

    2007-10-09

    The present state of the theory of fluctuations in gyrokinetic GK plasmas and especially its application to sampling noise in GK particle-in-cell PIC simulations is reviewed. Topics addressed include the Δf method, the fluctuation-dissipation theorem for both classical and GK many-body plasmas, the Klimontovich formalism, sampling noise in PIC simulations, statistical closure for partial differential equations, the theoretical foundations of spectral balance in the presence of arbitrary noise sources, and the derivation of Kadomtsev-type equations from the general formalism.

  20. FoxO1 and HNF-4 Are Involved in Regulation of Hepatic Glucokinase Gene Expression by Resveratrol*

    Science.gov (United States)

    Ganjam, Goutham Kumar; Dimova, Elitsa Y.; Unterman, Terry G.; Kietzmann, Thomas

    2009-01-01

    Resveratrol, a polyphenol derived from grapes, exerts important effects on glucose and lipid metabolism, yet detailed mechanisms mediating these effects remain unknown. The liver plays a central role in energy homeostasis, and glucokinase (GK) is a key enzyme involved in glucose utilization. Resveratrol activates SIRT1 (sirtuin 1), which promotes deacetylation of the forkhead transcription factor FoxO1. Previously, we reported that FoxO1 can suppress and that HNF-4 can stimulate GK expression in the liver. Here, we examined the role of FoxO1 and HNF-4 in mediating resveratrol effects on liver GK expression. Resveratrol suppressed hepatic GK expression in vivo and in isolated hepatocytes, and knocking down FoxO1 with shRNAs disrupted this effect. Reporter gene, gel shift, supershift assay, and chromatin immunoprecipitation studies show that FoxO1 binds to the GK promoter and that the interplay between FoxO1 and HNF-4 within the GK promoter is essential for mediating the effects of resveratrol. Resveratrol promotes deacetylation of FoxO1 and enhances its recruitment to the FoxO-binding element. Conversely, resveratrol suppresses recruitment of HNF-4 to its binding site, and knockdown of FoxO1 blocks this effect of resveratrol. Coprecipitation and chromatin immunoprecipitation studies show that resveratrol enhances interaction between FoxO1 and HNF-4, reduces binding of HNF-4 to its own site, and promotes its recruitment to the FoxO site in a FoxO1-dependent manner. These results provide the first evidence that resveratrol represses GK expression via FoxO1 and that the interaction between FoxO1 and HNF-4 contributes to these effects of resveratrol. PMID:19740748

  1. Food-intake dysregulation in type 2 diabetic Goto-Kakizaki rats: hypothesized role of dysfunctional brainstem thyrotropin-releasing hormone and impaired vagal output.

    Science.gov (United States)

    Zhao, K; Ao, Y; Harper, R M; Go, V L W; Yang, H

    2013-09-05

    Thyrotropin-releasing hormone (TRH), a neuropeptide contained in neural terminals innervating brainstem vagal motor neurons, enhances vagal outflow to modify multisystemic visceral functions and food intake. Type 2 diabetes (T2D) and obesity are accompanied by impaired vagal functioning. We examined the possibility that impaired brainstem TRH action may contribute to the vagal dysregulation of food intake in Goto-Kakizaki (GK) rats, a T2D model with hyperglycemia and impaired central vagal activation by TRH. Food intake induced by intracisternal injection of TRH analog was reduced significantly by 50% in GK rats, compared to Wistar rats. Similarly, natural food intake in the dark phase or food intake after an overnight fast was reduced by 56-81% in GK rats. Fasting (48h) and refeeding (2h)-associated changes in serum ghrelin, insulin, peptide YY, pancreatic polypeptide and leptin, and the concomitant changes in orexigenic or anorexigenic peptide expression in the brainstem and hypothalamus, all apparent in Wistar rats, were absent or markedly reduced in GK rats, with hormone release stimulated by vagal activation, such as ghrelin and pancreatic polypeptide, decreased substantially. Fasting-induced Fos expression accompanying endogenous brainstem TRH action decreased by 66% and 91%, respectively, in the nucleus tractus solitarius (NTS) and the dorsal motor nucleus of the vagus (DMV) in GK rats, compared to Wistar rats. Refeeding abolished fasting-induced Fos-expression in the NTS, while that in the DMV remained in Wistar but not GK rats. These findings indicate that dysfunctional brainstem TRH-elicited vagal impairment contributes to the disturbed food intake in T2D GK rats, and may provide a pathophysiological mechanism which prevents further weight gain in T2D and obesity. Published by Elsevier Ltd.

  2. Effect of modified atmosphere packaging on quality and shelf life of 'Robusta' banana (Musa sp.) stored at low temperature.

    Science.gov (United States)

    Kudachikar, V B; Kulkarni, S G; Prakash, M N Keshava

    2011-06-01

    Banana (Musa sp var. 'Robusta') stored under active and passive modified atmosphere packaging (MAP) at 12 ± 1°C and 85-90% RH for 2 seasons were evaluated for fruit quality and shelf-life. A steady state of about 8.6 and 8.2% of CO2 and 2.8 and 2.6% of O2 in passive MAP and MAP+GK (Green Keeper) packages, respectively, were established after 3 weeks of storage. Passive MAP and MAP+GK treatments of banana resulted in reduction in physiological loss in weight (PLW) of 0.7 and 0.8% after 5 and 7 weeks of storage, respectively as against 5% PLW in openly kept green banana after 3 weeks. Both MAP and MAP+GK treatments delayed colour, texture, pulp to peel ratio and total soluble solids (TSS) content as compared to openly kept control banana. Results indicated that the shelf life of fruits packed under MAP and MAP+GK could be extended up to 5 and 7 weeks, respectively as compared to 3 weeks for openly kept control fruits. Sensory quality of fully ripe fruits of both passive MAP and MAP+GK treatments, 5 days after ethrel dip was very good. Thus, MAP+GK at 12 ± 1°C and 85-90% RH could be commercially used for long term storage and long distance transportation of banana with maximum shelf-life of 7 weeks.

  3. Lipasin/betatrophin is differentially expressed in liver and white adipose tissue without association with insulin resistance in Wistar and Goto-Kakizaki rats.

    Science.gov (United States)

    Cahová, M; Habart, D; Olejár, T; Berková, Z; Papáčková, Z; Daňková, H; Lodererova, A; Heczková, M; Saudek, F

    2017-05-04

    Lipasin is a recently identified lipokine expressed predominantly in liver and in adipose tissue. It was linked to insulin resistance in mice and to type 1 and type 2 diabetes (T1D, T2D) in humans. No metabolic studies concerning lipasin were performed yet in rats. Therefore, we used rat model of T2D and insulin resistance, Goto-Kakizaki (GK) rats, to determine changes of lipasin expression in liver and in white adipose tissue (WAT) over 52 weeks in the relation to glucose tolerance, peripheral tissue insulin sensitivity and adiposity. GK rats were grossly glucose intolerant since the age of 6 weeks and developed peripheral insulin resistance at the age of 20 weeks. Expression of lipasin in the liver did not differ between GK and Wistar rats, declining with age, and it was not related to hepatic triacylglycerol content. In WAT, the lipasin expression was significantly higher in Wistar rats where it correlated positively with adiposity. No such correlation was found in GK rats. In conclusion, lipasin expression was associated neither with a mild age-related insulin resistance (Wistar), nor with severe genetically-based insulin resistance (GK).

  4. Synthesis of 24S and 24R-hydroxy-[24-3H] vitamin D3 and their metabolism in rachitic rats

    International Nuclear Information System (INIS)

    Tanaka, Y.; DeLuca, H.F.; Akaiwa, A.; Morisaki, M.; Ikekawa, N.

    1976-01-01

    An epimeric mixture of 24-hydroxy-[24- 3 H] vitamin D 3 was synthesized by the reduction of 24-ketovitamin D 3 by sodium borotritide. The epimeric mixture was converted to the trimethylsilylether derivatives and subjected to high-pressure liquid chromatography using silica gel columns to separate the 24-hydroxy-[24- 3 H] vitamin D 3 isomers. The 24R-hydroxy-[24- 3 H]vitamin D 3 induced calcification in rachitic rats while the 24S-hydroxy-[24- 3 H]vitamin D 3 had little or no such activity. As both isomers of 24-hydroxy-vitamin D 3 are metabolized to 24,25-dihydroxyvitamin D 3 , it appears that the 24-hydroxyvitamin D 3 -25-hydroxylase does not discriminate between the isomers. Only the R-isomer of 24-hydroxyvitamin D 3 is metabolized to 1,24-dihydroxyvitamin D 3 , although only trace amounts of this compound were found 2 days after the administration of 24-hydroxyvitamin D 3 . The striking difference in the metabolism of the isomers is the high selectivity of the 1-hydroxylase for the R-isomer. It is suggested that the high specificity of biological activity for the R-isomer of 24-hydroxyvitamin D 3 is because of the specificity of the 1-hydroxylation of 24,25-dihydroxyvitamin D 3 for the R configuration

  5. Differences in strength and speed demands between 4v4 and 8v8 small-sided football games

    DEFF Research Database (Denmark)

    Rebelo, António Natal Campos; Silva, Pedro; Rago, Vincenzo

    2016-01-01

    The aims of this study were (i) to characterise the acceleration demands of two different formats of small-sided game (SSG), i.e., 4v4 + goalkeepers (4v4 + GK) and 8v8 + goalkeepers (8v8 + GK); (ii) to analyse the correlation between performance in power-based tests and acceleration-based physica...... to repetitions and fatigue development of muscle power-based actions than 8v8 + GK. It may therefore be logical to use the former type of SSG to target development of power-related football actions....

  6. Incorporating climate change and technology into the science classroom: Lessons from my year as a GK-12 Fellow

    Science.gov (United States)

    Abramoff, R. Z.

    2012-12-01

    Climate change is not included in the K-8 science standards in Massachusetts; as a result, students learn what climate is, but not how human activities affect it. Starting in 2010, Boston University launched the GK-12 GLACIER program, funded with 2.9M from the National Science Foundation. The purpose of the program is to incorporate the fundamentals of climate change into the K-12 curriculum, focusing on grades 5-8 when quantitative science enters the curriculum. Graduate students are partnered with teachers in Boston public schools for 10 hours a week of teaching with additional curriculum development. I will focus on the curriculum that I developed as a part of this program for the 5th grade science class at The Curley School in Jamaica Plain, MA, where I worked with Grades 3-5, ESL, and PACE autism program science teacher, Stephanie Selznick. The Curley School is an ethnically and economically diverse Boston public school with about 800 students and an 83% minority population. At the Curley, I taught two full days a week, meeting with all of the 5th grade classes and some of the 4th grade classes of all academic levels. The lessons that I created were designed to fit into the state standards and enrich student understanding plant ecology and earth science, as well as develop their capacity to design experiments and use technology. These include Question of the Day, Digital Field Guide to the Outdoor Classroom, Phototropism, Solar System Weather Report, Soil and Water, Local Landforms, and the Earth as a Closed System Unit for which materials and lesson plans are available on my website. Our secondary goals were to improve tech literacy at Curley. Due to funding restrictions, there were few technology resources available to the students at the beginning of the 2011/2012 school year. To improve technology resources at Curley, I organized a fundraiser at Boston University, selling donated items from graduate students and faculty; the 1000 raised was used to supply

  7. 33 CFR 83.24 - Towing and pushing (Rule 24).

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Towing and pushing (Rule 24). 83... NAVIGATION RULES RULES Lights and Shapes § 83.24 Towing and pushing (Rule 24). (a) A power-driven vessel when... it can best be seen. (b) Pushing vessel and pushed vessel rigidly connected in composite unit. When a...

  8. Preparation and characterization of gum karaya hydrogel nanocomposite flocculant for metal ions removal from mine effluents

    CSIR Research Space (South Africa)

    Fosso-Kankeu, E

    2016-02-01

    Full Text Available This research paper reports the removal of heavy metal ions from mine effluents using the gum karaya (GK)-grafted poly(acrylamide-co-acrylic acid) incorporated iron oxide magnetic nanoparticles (Fe3O4 MNPs) hydrogel nanocomposite [i.e., GK...

  9. ???????? ??????? ????????? ?????????? ??????? ? ?????????? ?????? ??-24

    OpenAIRE

    ????????, ?????; ????????, ?????; ?????????, ?????; ??????, ??????

    2011-01-01

    ??????? ????????? ?????????????? ??????? ????????? ??? ?????????? ?????? ??-24 ?? ?????????? ???????? (??). ????????????? ???????? ??????? ????????? ?? ?????? ????? ????? ?? ???????, ?? ??????? ?????? ????????? ?? ???????????? ?????, ?? ?????????? ?????.

  10. The magnetic fields at the surface of active single G-K giants

    Science.gov (United States)

    Aurière, M.; Konstantinova-Antova, R.; Charbonnel, C.; Wade, G. A.; Tsvetkova, S.; Petit, P.; Dintrans, B.; Drake, N. A.; Decressin, T.; Lagarde, N.; Donati, J.-F.; Roudier, T.; Lignières, F.; Schröder, K.-P.; Landstreet, J. D.; Lèbre, A.; Weiss, W. W.; Zahn, J.-P.

    2015-02-01

    Aims: We investigate the magnetic field at the surface of 48 red giants selected as promising for detection of Stokes V Zeeman signatures in their spectral lines. In our sample, 24 stars are identified from the literature as presenting moderate to strong signs of magnetic activity. An additional 7 stars are identified as those in which thermohaline mixing appears not to have occured, which could be due to hosting a strong magnetic field. Finally, we observed 17 additional very bright stars which enable a sensitive search to be performed with the spectropolarimetric technique. Methods: We use the spectropolarimeters Narval and ESPaDOnS to detect circular polarization within the photospheric absorption lines of our targets. We treat the spectropolarimetric data using the least-squares deconvolution method to create high signal-to-noise ratio mean Stokes V profiles. We also measure the classical S-index activity indicator for the Ca ii H&K lines, and the stellar radial velocity. To infer the evolutionary status of our giants and to interpret our results, we use state-of-the-art stellar evolutionary models with predictions of convective turnover times. Results: We unambiguously detect magnetic fields via Zeeman signatures in 29 of the 48 red giants in our sample. Zeeman signatures are found in all but one of the 24 red giants exhibiting signs of activity, as well as 6 out of 17 bright giant stars. However no detections were obtained in the 7 thermohaline deviant giants. The majority of the magnetically detected giants are either in the first dredge up phase or at the beginning of core He burning, i.e. phases when the convective turnover time is at a maximum: this corresponds to a "magnetic strip" for red giants in the Hertzsprung-Russell diagram. A close study of the 16 giants with known rotational periods shows that the measured magnetic field strength is tightly correlated with the rotational properties, namely to the rotational period and to the Rossby number Ro

  11. Deoxyribonucleoside kinases in mitochondrial DNA depletion.

    Science.gov (United States)

    Saada-Reisch, Ann

    2004-10-01

    Mitochondrial DNA (mtDNA) depletion syndromes (MDS) are a heterogeneous group of mitochondrial disorders, manifested by a decreased mtDNA copy number and respiratory chain dysfunction. Primary MDS are inherited autosomally and may affect a single organ or multiple tissues. Mutated mitochondrial deoxyribonucleoside kinases; deoxyguanosine kinase (dGK) and thymidine kinase 2 (TK2), were associated with the hepatocerebral and myopathic forms of MDS respectively. dGK and TK2 are key enzymes in the mitochondrial nucleotide salvage pathway, providing the mitochondria with deoxyribonucleotides (dNP) essential for mtDNA synthesis. Although the mitochondrial dNP pool is physically separated from the cytosolic one, dNP's may still be imported through specific transport. Non-replicating tissues, where cytosolic dNP supply is down regulated, are thus particularly vulnerable to dGK and TK2 deficiency. The overlapping substrate specificity of deoxycytidine kinase (dCK) may explain the relative sparing of muscle in dGK deficiency, while low basal TK2 activity render this tissue susceptible to TK2 deficiency. The precise pathophysiological mechanisms of mtDNA depletion due to dGK and TK2 deficiencies remain to be determined, though recent findings confirm that it is attributed to imbalanced dNTP pools.

  12. PGC-1α mRNA Level and Oxidative Capacity of the Plantaris Muscle in Rats with Metabolic Syndrome, Hypertension, and Type 2 Diabetes

    International Nuclear Information System (INIS)

    Nagatomo, Fumiko; Fujino, Hidemi; Kondo, Hiroyo; Gu, Ning; Takeda, Isao; Ishioka, Noriaki; Tsuda, Kinsuke; Ishihara, Akihiko

    2011-01-01

    We examined the fiber profiles and the mRNA levels of peroxisome proliferator-activated receptors (PPARα and PPARδ/β) and of the PPARγ coactivator-1α (PGC-1α) in the plantaris muscles of 15-week-old control (WR), metabolic syndrome (CP), hypertensive (SHR), and type 2 diabetic (GK) rats. The deep regions in the muscles of SHR and GK rats exhibited lower percentages of high-oxidative type I and IIA fibers and higher percentages of low-oxidative type IIB fibers compared with WR and CP rats. The surface regions in the muscles of CP, SHR, and GK rats exhibited lower percentages of high-oxidative type IIA fibers and higher percentages of low-oxidative type IIB fibers compared with WR rats. The muscles of SHR and GK rats had lower oxidative enzyme activity compared with WR rats. The muscles of SHR rats had the lowest PPARδ/β mRNA level. In addition, the muscles of SHR and GK rats had lower PGC-1α mRNA level compared with WR and CP rats. We concluded that the plantaris muscles of rats with hypertension and type 2 diabetes have lower oxidative capacity, which is associated with the decreased level of PGC-1α mRNA

  13. A review of the microbiology of the Rehai geothermal field in Tengchong, Yunnan Province, China

    Directory of Open Access Journals (Sweden)

    Brian P. Hedlund

    2012-05-01

    Full Text Available The Rehai Geothermal Field, located in Tengchong County, in central-western Yunnan Province, is the largest and most intensively studied geothermal field in China. A wide physicochemical diversity of springs (ambient to ∼97 °C; pH from ≤1.8 to ≥9.3 provides a multitude of niches for extremophilic microorganisms. A variety of studies have focused on the cultivation, identification, basic physiology, taxonomy, and biotechnological potential of thermophilic microorganisms from Rehai. Thermophilic bacteria isolated from Rehai belong to the phyla Firmicutes and Deinococcus-Thermus. Firmicutes include neutrophilic or alkaliphilic Anoxybacillus, Bacillus, Caldalkalibacillus, Caldanaerobacter, Laceyella, and Geobacillus, as well as thermoacidophilic Alicyclobacillus and Sulfobacillus. Isolates from the Deinococcus-Thermus phylum include several Meiothermus and Thermus species. Many of these bacteria synthesize thermostable polymer-degrading enzymes that may be useful for biotechnology. The thermoacidophilic archaea Acidianus, Metallosphaera, and Sulfolobus have also been isolated and studied. A few studies have reported the isolation of thermophilic viruses belonging to Siphoviridae (TTSP4 and TTSP10 and Fuselloviridae (STSV1 infecting Thermus spp. and Sulfolobus spp., respectively. More recently, cultivation-independent studies using 16S rRNA gene sequences, shotgun metagenomics, or “functional gene” sequences have revealed a much broader diversity of microorganisms than represented in culture. Studies of the gene and mRNA encoding the large subunit of the ammonia monooxygenase (amoA of ammonia-oxidizing Archaea (AOA and the tetraether lipid crenarchaeol, a potential biomarker for AOA, suggest a wide diversity, but possibly low abundance, of thermophilic AOA in Rehai. Finally, we introduce the Tengchong Partnerships in International Research and Education (PIRE project, an international collaboration between Chinese and U.S. scientists with

  14. Overproduction, purification, crystallization and preliminary X-ray diffraction analysis of Trypanosoma brucei gambiense glycerol kinase

    International Nuclear Information System (INIS)

    Balogun, Emmanuel Oluwadare; Inaoka, Daniel Ken; Kido, Yasutoshi; Shiba, Tomoo; Nara, Takeshi; Aoki, Takashi; Honma, Teruki; Tanaka, Akiko; Inoue, Masayuki; Matsuoka, Shigeru; Michels, Paul A. M.; Harada, Shigeharu; Kita, Kiyoshi

    2010-01-01

    Glycerol kinase from human African trypanosomes has been purified and crystallized for X-ray structure analysis. In the bloodstream forms of human trypanosomes, glycerol kinase (GK; EC 2.7.1.30) is one of the nine glycosomally compartmentalized enzymes that are essential for energy metabolism. In this study, a recombinant Trypanosoma brucei gambiense GK (rTbgGK) with an N-terminal cleavable His 6 tag was overexpressed, purified to homogeneity and crystallized by the sitting-drop vapour-diffusion method using PEG 400 as a precipitant. A complete X-ray diffraction data set to 2.75 Å resolution indicated that the crystals belonged to the orthorhombic space group P2 1 2 1 2 1 , with unit-cell parameters a = 63.84, b = 121.50, c = 154.59 Å. The presence of two rTbgGK molecules in the asymmetric unit gives a Matthews coefficient (V M ) of 2.5 Å 3 Da −1 , corresponding to 50% solvent content

  15. WE-G-BRA-08: Failure Modes and Effects Analysis (FMEA) for Gamma Knife Radiosurgery

    International Nuclear Information System (INIS)

    Xu, Y; Bhatnagar, J; Bednarz, G; Flickinger, J; Arai, Y; Huq, M Saiful; Vacsulka, J; Monaco, E; Niranjan, A; Lunsford, L Dade; Feng, W

    2015-01-01

    Purpose: To perform a failure modes and effects analysis (FMEA) study for Gamma Knife (GK) radiosurgery processes at our institution based on our experience with the treatment of more than 13,000 patients. Methods: A team consisting of medical physicists, nurses, radiation oncologists, neurosurgeons at the University of Pittsburgh Medical Center and an external physicist expert was formed for the FMEA study. A process tree and a failure mode table were created for the GK procedures using the Leksell GK Perfexion and 4C units. Three scores for the probability of occurrence (O), the severity (S), and the probability of no detection (D) for failure modes were assigned to each failure mode by each professional on a scale from 1 to 10. The risk priority number (RPN) for each failure mode was then calculated (RPN = OxSxD) as the average scores from all data sets collected. Results: The established process tree for GK radiosurgery consists of 10 sub-processes and 53 steps, including a sub-process for frame placement and 11 steps that are directly related to the frame-based nature of the GK radiosurgery. Out of the 86 failure modes identified, 40 failure modes are GK specific, caused by the potential for inappropriate use of the radiosurgery head frame, the imaging fiducial boxes, the GK helmets and plugs, and the GammaPlan treatment planning system. The other 46 failure modes are associated with the registration, imaging, image transfer, contouring processes that are common for all radiation therapy techniques. The failure modes with the highest hazard scores are related to imperfect frame adaptor attachment, bad fiducial box assembly, overlooked target areas, inaccurate previous treatment information and excessive patient movement during MRI scan. Conclusion: The implementation of the FMEA approach for Gamma Knife radiosurgery enabled deeper understanding of the overall process among all professionals involved in the care of the patient and helped identify potential

  16. Interleukin-1/toll-like receptor-induced nuclear factor kappa B signaling participates in intima hyperplasia after carotid artery balloon injury in goto-kakizaki rats: a potential target therapy pathway.

    Directory of Open Access Journals (Sweden)

    Xiaotian Zhang

    Full Text Available The value of restenosis after percutaneous coronary intervention (PCI is recognized worldwide, especially for diabetic patients. Interleukin-1/Toll-like receptor (IL-1/TLR signaling is involved in innate and adaptive immune responses, but whether and how the IL-1/TLR-induced nuclear factor kappa B (NFκB pathway plays key roles in intimal formation is unclear. The underlying mechanism of intima hyperplasia was investigated with a model of carotid balloon injury in Goto-Kakizaki (GK and Wistar rats and with lipopolysaccharide-stimulated macrophages. Elastic-van Gieson staining showed the medial area peakedon Day 3 post-injury and decreased by Day 7 post-injury in both GK and Wistar rats. The N/M at Day 7 in GK rats was significantly higher than in Wistar rats (p<0.001. The percent of 5-ethynyl-2'-deoxyuridine (EdU staining-positive cells on Day 3 post-injury was greater than seen on Day 7 post-injury in GK and Wistar rats. The percent of EdU-positive cells on Days 3 and 7 post-injury in Wistar rats was less than that found in GK rats (p<0.01; p<0.05. NFκBp65 immunostaining had increased by Day 7 post-injury. Agilent Whole Genome Oligo Microarray verified that the IL-1/TLR-induced NFκB pathway was activated by carotid balloon injury. TLR4, IL-1 receptor associated kinase, inhibitors α of NFκB, human antigen R, c-Myc (Proto-Oncogene Proteins, EGF-like module-containing mucin-like hormone receptor-like 1 and Interleukin-6 were up-regulated or down-regulated according to immunochemistry, quantitative real-time PCR, Western blotting and Enzyme linked immunosorbent assay. Overall, we conclude that the IL-1/TLR-induced NFκB pathway participates in the intimal hyperplasia after carotid injury in GK and Wistar rats and that GK rats respond more intensely to the inflammation than Wistar rats.

  17. Rbm24a and Rbm24b are required for normal somitogenesis.

    Directory of Open Access Journals (Sweden)

    Samantha Maragh

    Full Text Available We recently demonstrated that the gene encoding the RNA binding motif protein 24 (RBM24 is expressed during mouse cardiogenesis, and determined the developmental requirement for its zebrafish homologs Rbm24a and Rbm24b during cardiac development. We demonstrate here that both Rbm24a and Rbm24b are also required for normal somite and craniofacial development. Diminution of rbm24a or rbm24b gene products by morpholino knockdown resulted in significant disruption of somite formation. Detailed in situ hybridization-based analyses of a spectrum of somitogenesis-associated transcripts revealed reduced expression of the cyclic muscle pattering genes dlc and dld encoding Notch ligands, as well as their respective target genes her7, her1. By contrast expression of the Notch receptors notch1a and notch3 appears unchanged. Some RBM-family members have been implicated in pre-mRNA processing. Analysis of affected Notch-pathway mRNAs in rbm24a and rbm24b morpholino-injected embryos revealed aberrant transcript fragments of dlc and dld, but not her1 or her7, suggesting the reduction in transcription levels of Notch pathway components may result from aberrant processing of its ligands. These data imply a previously unknown requirement for Rbm24a and Rbm24b in somite and craniofacial development. Although we anticipate the influence of disrupting RBM24 homologs likely extends beyond the Notch pathway, our results suggest their perturbation may directly, or indirectly, compromise post-transcriptional processing, exemplified by imprecise processing of dlc and dld.

  18. Effects of α-lipoic acid on endothelial function in aged diabetic and high-fat fed rats

    Science.gov (United States)

    Sena, C M; Nunes, E; Louro, T; Proença, T; Fernandes, R; Boarder, M R; Seiça, R M

    2007-01-01

    Background and purpose: This study was conducted to investigate the effects of α-lipoic acid (α-LA) on endothelial function in diabetic and high-fat fed animal models and elucidate the potential mechanism underlying the benefits of α-LA. Experimental approach: Plasma metabolites reflecting glucose and lipid metabolism, endothelial function, urinary albumin excretion (UAE), plasma and aortic malondialdehyde (MDA) and urinary 8-hydroxydeoxyguanosine (8-OHdG) were assessed in non-diabetic controls (Wistar rats), untreated Goto-Kakizaki (GK) diabetic and high-fat fed GK rats (fed with atherogenic diet only, treated with α-LA and treated with vehicle, for 3 months). Vascular eNOS, nitrotyrosine, carbonyl groups and superoxide anion were also assessed in the different groups. Key results: α-LA and soybean oil significantly reduced both total and non-HDL serum cholesterol and triglycerides induced by atherogenic diet. MDA, carbonyl groups, vascular superoxide and 8-OHdG levels were higher in GK and high-fat fed GK groups and fully reversed with α-LA treatment. High-fat fed GK diabetic rats showed significantly reduced endothelial function and increased UAE, effects ameliorated with α-LA. This endothelial dysfunction was associated with decreased NO production, decreased expression of eNOS and increased vascular superoxide production and nitrotyrosine expression. Conclusions and implications: α-LA restores endothelial function and significantly improves systemic and local oxidative stress in high-fat fed GK diabetic rats. Improved endothelial function due to α-LA was at least partially attributed to recoupling of eNOS and increased NO bioavailability and represents a pharmacological approach to prevent major complications associated with type 2 diabetes. PMID:17906683

  19. Molecular mechanisms of mitochondrial DNA depletion diseases caused by deficiencies in enzymes in purine and pyrimidine metabolism.

    Science.gov (United States)

    Eriksson, Staffan; Wang, Liya

    2008-06-01

    Mitochondrial DNA depletion syndrome (MDS), a reduction of mitochondrial DNA copy number, often affects muscle or liver. Mutations in enzymes of deoxyribonucleotide metabolism give MDS, for example, the mitochondrial thymidine kinase 2 (TK2) and deoxyguanosine kinase (dGK) genes. Sixteen TK2 and 22 dGK alterations are known. Their characteristics and symptoms are described. Levels of five key deoxynucleotide metabolizing enzymes in mouse tissues were measured. TK2 and dGK levels in muscles were 5- to 10-fold lower than other nonproliferating tissues and 100-fold lower compared to spleen. Each type of tissue apparently relies on de novo and salvage synthesis of DNA precursors to varying degrees.

  20. Glucosensing capacity in rainbow trout liver displays day-night variations possibly related to melatonin action.

    Science.gov (United States)

    Conde-Sieira, Marta; Patiño, Marcos A López; Míguez, Jesús M; Soengas, José L

    2012-09-01

    To assess whether the glucosensing capacity in peripheral (liver and Brockmann bodies) and central (hypothalamus and hindbrain) locations of rainbow trout displays day-night variations in its response to changes in circulating glucose levels, we evaluated the response of parameters related to glucosensing [glucose, glycogen and glucose 6-phosphate levels, activities of glucokinase (GK), glycogen synthetase (GSase) and pyruvate kinase (PK), and mRNA abundance of GK, glucose transporter 2 (GLUT2), and K(ATP) channel subunits Kir6.x-like and sulfonylurea receptor (SUR)-like] in fish subjected to hyperglycemic treatment under night or day conditions. No day-night significant variations were noticed in the glucosensing capacity of the hypothalamus, hindbrain and Brockmann bodies. In contrast, a clear differential response was noticed in the liver, where glucose levels, GK activity (and mRNA levels) and GSase activity displayed increased values during the day in hyperglycemic fish compared with controls, and lower (GK mRNA levels) or non-existent (glucose, GK and GSase activities, and Kir6.x-like mRNA levels) values during the night. A similar decrease in parameters related to glucosensing in the liver was observed when fish under day conditions were treated with melatonin, suggesting a modulatory role of melatonin in day-night changes of the glucosensing response in the same tissue.

  1. Yeast artificial chromosome cloning in the glycerol kinase and adrenal hypoplasia congenita region of Xp21

    Energy Technology Data Exchange (ETDEWEB)

    Worley, K.C.; Ellison, K.A.; Zhang, Y.H.; Wang, D.F.; Mason, J.; Roth, E.J.; Adams, V.; Fogt, D.D.; Zhu, X.M.; Towbin, J.A. [Baylor College of Medicine, Houston, TX (United States)] [and others

    1993-05-01

    The adrenal hypoplasia congenita (AHC) and glycerol kinase (GK) loci are telomeric to the Duchenne muscular dystrophy locus in Xp21. The authors developed a pair of yeast artificial chromosome (YAC) contigs spanning at least 1.2 Mb and encompassing the region from the telomeric end of the Duchenne muscular dystrophy (DMD) locus to beyond YHX39 (DXS727), including the genes for AHC and GK. The centromeric contig consists of 13 YACs reaching more than 600 kb from DMD through GK. The telomeric contig group consists of 8 YACs containing more than 600 kb including the markers YHX39 (DXS727) and QST-59 (DXS319). Patient deletion breakpoints in the region of the two YAC contigs define at least eight intervals, and seven deletion breakpoints are contained within these contigs. In addition to the probes developed from YAC ends, they have mapped eight Alu-PCR probes amplified from a radiation-reduced somatic cell hybrid, two anonymous DNA probes, and one Alu-PCR product amplified from a cosmid end, for a total of 26 new markers within this region of 2 Mb or less. One YAC in the centromeric contig contains an insert encompassing the minimum interval for GK deficiency defined by patient deletion breakpoints, and this clone includes all or part of the GK gene. 33 refs., 3 figs., 5 tabs.

  2. Thermonuclear 19F(p, {{\\boldsymbol{\\alpha }}}_{0})16O reaction rate

    Science.gov (United States)

    He, Jian-Jun; Lombardo, Ivano; Dell'Aquila, Daniele; Xu, Yi; Zhang, Li-Yong; Liu, Wei-Ping

    2018-01-01

    The thermonuclear 19F(p, {{{α }}}0)16O reaction rate in the temperature region 0.007-10 GK has been derived by re-evaluating the available experimental data, together with the low-energy theoretical R-matrix extrapolations. Our new rate deviates by up to about 30% compared to the previous results, although all rates are consistent within the uncertainties. At very low temperature (e.g. 0.01 GK) our reaction rate is about 20% lower than the most recently published rate, because of a difference in the low energy extrapolated S-factor and a more accurate estimate of the reduced mass used in the calculation of the reaction rate. At temperatures above ˜1 GK, our rate is lower, for instance, by about 20% around 1.75 GK, because we have re-evaluated the previous data (Isoya et al., Nucl. Phys. 7, 116 (1958)) in a meticulous way. The present interpretation is supported by the direct experimental data. The uncertainties of the present evaluated rate are estimated to be about 20% in the temperature region below 0.2 GK, and are mainly caused by the lack of low-energy experimental data and the large uncertainties in the existing data. Asymptotic giant branch (AGB) stars evolve at temperatures below 0.2 GK, where the 19F(p, {{α }})16O reaction may play a very important role. However, the current accuracy of the reaction rate is insufficient to help to describe, in a careful way, the fluorine over-abundances observed in AGB stars. Precise cross section (or S factor) data in the low energy region are therefore needed for astrophysical nucleosynthesis studies. Supported by National Natural Science Foundation of China (11490562, 11490560, 11675229) and National Key Research and Development Program of China (2016YFA0400503)

  3. Synergistic Interaction of Hypertension and Diabetes in Promoting Kidney Injury and the Role of Endoplasmic Reticulum Stress.

    Science.gov (United States)

    Wang, Zhen; do Carmo, Jussara M; Aberdein, Nicola; Zhou, Xinchun; Williams, Jan M; da Silva, Alexandre A; Hall, John E

    2017-05-01

    Diabetes mellitus and hypertension are major risk factors for chronic kidney injury, together accounting for >70% of end-stage renal disease. In this study, we assessed interactions of hypertension and diabetes mellitus in causing kidney dysfunction and injury and the role of endoplasmic reticulum (ER) stress. Hypertension was induced by aorta constriction (AC) between the renal arteries in 6-month-old male Goto-Kakizaki (GK) type 2 diabetic and control Wistar rats. Fasting plasma glucose averaged 162±11 and 87±2 mg/dL in GK and Wistar rats, respectively. AC produced hypertension in the right kidney (above AC) and near normal blood pressure in the left kidney (below AC), with both kidneys exposed to the same levels of glucose, circulating hormones, and neural influences. After 8 weeks of AC, blood pressure above the AC (and in the right kidney) increased from 109±1 to 152±5 mm Hg in GK rats and from 106±4 to 141±5 mm Hg in Wistar rats. The diabetic-hypertensive right kidneys in GK-AC rats had much greater increases in albumin excretion and histological injury compared with left kidneys (diabetes mellitus only) of GK rats or right kidneys (hypertension only) of Wistar-AC rats. Marked increases in ER stress and oxidative stress indicators were observed in diabetic-hypertensive kidneys of GK-AC rats. Inhibition of ER stress with tauroursodeoxycholic acid for 6 weeks reduced blood pressure (135±4 versus 151±4 mm Hg), albumin excretion, ER and oxidative stress, and glomerular injury, while increasing glomerular filtration rate in hypertensive-diabetic kidneys. These results suggest that diabetes mellitus and hypertension interact synergistically to promote kidney dysfunction and injury via ER stress. © 2017 American Heart Association, Inc.

  4. Diabetic beta-cells can achieve self-protection against oxidative stress through an adaptive up-regulation of their antioxidant defenses.

    Directory of Open Access Journals (Sweden)

    Grégory Lacraz

    Full Text Available BACKGROUND: Oxidative stress (OS, through excessive and/or chronic reactive oxygen species (ROS, is a mediator of diabetes-related damages in various tissues including pancreatic beta-cells. Here, we have evaluated islet OS status and beta-cell response to ROS using the GK/Par rat as a model of type 2 diabetes. METHODOLOGY/PRINCIPAL FINDINGS: Localization of OS markers was performed on whole pancreases. Using islets isolated from 7-day-old or 2.5-month-old male GK/Par and Wistar control rats, 1 gene expression was analyzed by qRT-PCR; 2 insulin secretion rate was measured; 3 ROS accumulation and mitochondrial polarization were assessed by fluorescence methods; 4 antioxidant contents were quantified by HPLC. After diabetes onset, OS markers targeted mostly peri-islet vascular and inflammatory areas, and not islet cells. GK/Par islets revealed in fact protected against OS, because they maintained basal ROS accumulation similar or even lower than Wistar islets. Remarkably, GK/Par insulin secretion also exhibited strong resistance to the toxic effect of exogenous H(2O(2 or endogenous ROS exposure. Such adaptation was associated to both high glutathione content and overexpression (mRNA and/or protein levels of a large set of genes encoding antioxidant proteins as well as UCP2. Finally, we showed that such a phenotype was not innate but spontaneously acquired after diabetes onset, as the result of an adaptive response to the diabetic environment. CONCLUSIONS: The GK/Par model illustrates the effectiveness of adaptive response to OS by beta-cells to achieve self-tolerance. It remains to be determined to what extend such islet antioxidant defenses upregulation might contribute to GK/Par beta-cell secretory dysfunction.

  5. Bovine oocytes and early embryos express mRNA encoding glycerol kinase but addition of glycerol to the culture media interferes with oocyte maturation.

    Science.gov (United States)

    Okawara, Sumika; Hamano, Seizo; Tetsuka, Masafumi

    2009-04-01

    Glycerol plays multi-functional roles in cellular physiology. Other than forming the backbone molecule for glycerophospholipid and triglyceride (TG), glycerol acts as an energy substrate for glycolysis. Spermatozoa are known to utilize glycerol for energy production, but there are no reports of this in oocytes. In this study, the value of glycerol as an energy substrate for bovine oocyte maturation (Exp. 1) and the gene expression of glycerol kinase (GK), an enzyme crucial for cellular glycerol utilization, in bovine oocytes and early embryos (Exp. 2) were examined. In Exp. 1, in vitro maturation (IVM) was conducted using synthetic oviduct fluid supplemented with/without glucose (1.5 mM) and/or glycerol (1.0 mM), and maturation rate, degree of cumulus expansion, glucose consumption and lactate production by cumulus-oocyte complexes (COC) were examined. In Exp. 2, to examine the developmental expression of GK mRNA, cumulus cells, oocytes and embryos at the 2-, 8- and 16-cell, morula, expanded blastocyst and hatched blastocyst stages were obtained in separate experiments, and the expression of GK mRNA was quantified using a real-time PCR. Glycerol did not support oocyte maturation or cumulus expansion. Addition of glycerol to glucose-supplemented media significantly decreased the maturation rate. Expression of GK mRNA was very low in cumulus cells, whereas an appreciable level of the transcript was observed in the oocytes. GK mRNA was detected in embryos at all the stages examined, and its expression significantly increased at the morula stage. These results indicate that glycerol, at least at the present concentration, is not beneficial as a constituent of the medium for bovine oocyte maturation. However, the appreciable levels of GK mRNA found in the oocyte and embryo imply a physiological role for glycerol in bovine oocyte maturation and embryo development.

  6. Therapeutic Applications of Interleukin 24 (IL24): A Review ...

    African Journals Online (AJOL)

    IL24 has growth suppressive properties in a wide variety of human cancer cell lines without inducing harmful effects in normal cells. This review is focused on the role of IL 24 on tumor cell biology and its potential therapeutic applications. Keywords: Melanoma differentiation, Protein, Therapeutics, Interleukin, ...

  7. ORF Alignment: NC_005835 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ine pdb|1FJG|S Chain S, Structure ... Of The Thermus Thermophilus 30s Ribosomal Subunit In ... Complex With The Antibioti...cs Streptomycin, ... Spectinomycin, And Paromomycin ... Length = 84 ... Q

  8. ORF Alignment: NC_006461 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ine pdb|1FJG|S Chain S, Structure ... Of The Thermus Thermophilus 30s Ribosomal Subunit In ... Complex With The Antibioti...cs Streptomycin, ... Spectinomycin, And Paromomycin ... Length = 84 ... Q

  9. Lactose metabolism in Streptococcus lactis: studies with a mutant lacking glucokinase and mannose-phosphotransferase activities

    International Nuclear Information System (INIS)

    Thompson, J.; Chassy, B.M.; Egan, W.

    1985-01-01

    A mutant of Streptococcus lactis 133 has been isolated that lacks both glucokinase and phosphoenolpyruvate-dependent mannose- phosphotransferase (mannose-PTS) activities. The double mutant S. lactis 133 mannose-PTSd GK- is unable to utilize either exogenously supplied or intracellularly generated glucose for growth. Fluorographic analyses of metabolites formed during the metabolism of [ 14 C]lactose labeled specifically in the glucose or galactosyl moiety established that the cells were unable to phosphorylate intracellular glucose. However, cells of S. lactis 133 mannose-PTSd GK- readily metabolized intracellular glucose 6-phosphate, and the growth rates and cell yield of the mutant and parental strains on sucrose were the same. During growth on lactose, S. lactis 133 mannose-PTSd GK- fermented only the galactose moiety of the disaccharide, and 1 mol of glucose was generated per mol of lactose consumed. For an equivalent concentration of lactose, the cell yield of the mutant was 50% that of the wild type. The specific rate of lactose utilization by growing cells of S. lactis 133 mannose-PTSd GK- was ca. 50% greater than that of the wild type, but the cell doubling times were 70 and 47 min, respectively. High-resolution 31 P nuclear magnetic resonance studies of lactose transport by starved cells of S. lactis 133 and S. lactis 133 mannose-PTSd GK- showed that the latter cells contained elevated lactose-PTS activity. Throughout exponential growth on lactose, the mutant maintained an intracellular steady-state glucose concentration of 100 mM

  10. Lactose metabolism in Streptococcus lactis: studies with a mutant lacking glucokinase and mannose-phosphotransferase activities

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, J.; Chassy, B.M.; Egan, W.

    1985-04-01

    A mutant of Streptococcus lactis 133 has been isolated that lacks both glucokinase and phosphoenolpyruvate-dependent mannose- phosphotransferase (mannose-PTS) activities. The double mutant S. lactis 133 mannose-PTSd GK- is unable to utilize either exogenously supplied or intracellularly generated glucose for growth. Fluorographic analyses of metabolites formed during the metabolism of (/sup 14/C)lactose labeled specifically in the glucose or galactosyl moiety established that the cells were unable to phosphorylate intracellular glucose. However, cells of S. lactis 133 mannose-PTSd GK- readily metabolized intracellular glucose 6-phosphate, and the growth rates and cell yield of the mutant and parental strains on sucrose were the same. During growth on lactose, S. lactis 133 mannose-PTSd GK- fermented only the galactose moiety of the disaccharide, and 1 mol of glucose was generated per mol of lactose consumed. For an equivalent concentration of lactose, the cell yield of the mutant was 50% that of the wild type. The specific rate of lactose utilization by growing cells of S. lactis 133 mannose-PTSd GK- was ca. 50% greater than that of the wild type, but the cell doubling times were 70 and 47 min, respectively. High-resolution /sup 31/P nuclear magnetic resonance studies of lactose transport by starved cells of S. lactis 133 and S. lactis 133 mannose-PTSd GK- showed that the latter cells contained elevated lactose-PTS activity. Throughout exponential growth on lactose, the mutant maintained an intracellular steady-state glucose concentration of 100 mM.

  11. 3D Structure and Interaction of p24β and p24δ Golgi Dynamics Domains: Implication for p24 Complex Formation and Cargo Transport.

    Science.gov (United States)

    Nagae, Masamichi; Hirata, Tetsuya; Morita-Matsumoto, Kana; Theiler, Romina; Fujita, Morihisa; Kinoshita, Taroh; Yamaguchi, Yoshiki

    2016-10-09

    The p24 family consists of four subfamilies (p24α, p24β, p24γ, and p24δ), and the proteins are thought to form hetero-oligomeric complexes for efficient transport of cargo proteins from the endoplasmic reticulum to the Golgi apparatus. The proteins possess a conserved luminal Golgi dynamics (GOLD) domain, whose functions are largely unknown. Here, we present structural and biochemical studies of p24β1 and p24δ1 GOLD domains. Use of GOLD domain-deleted mutants revealed that the GOLD domain of p24δ1 is required for proper p24 hetero-oligomeric complex formation and efficient transport of GPI-anchored proteins. The p24β1 and p24δ1 GOLD domains share a common β-sandwich fold with a characteristic intrasheet disulfide bond. The GOLD domain of p24δ1 crystallized as dimers, allowing the analysis of a homophilic interaction site. Surface plasmon resonance and solution NMR analyses revealed that p24β1 and p24δ1 GOLD domains interact weakly (K d = ~10 -4 M). Bi-protein titration provided interaction site maps. We propose that the heterophilic interaction of p24 GOLD domains contributes to the formation of the p24 hetero-oligomeric complex and to efficient cargo transport. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. ASA24-Canada

    Science.gov (United States)

    A Canadian adaptation of the Automated Self-Administered 24-hour Dietary Assessment Tool (ASA24-Canada), developed by the Food Directorate at Health Canada in collaboration with NCI, has been freely available since April 2014.

  13. Basic studies on gastrin-radioimmunoassay and the results of its clinical application. Comparative studies between the double antibody method using Wilson's anti-gastrin serum and a gastrin kit (CIS) method

    Energy Technology Data Exchange (ETDEWEB)

    Yabana, T; Uchiya, T; Kakumoto, Y; Waga, Y; Konta, M [Sapporo Medical Coll. (Japan)

    1975-03-01

    Fundamental and practical problems in carrying out the radioimmunoassay of gastrin were studied by comparing the double antibody method, using guinea pig anti-porcine gastrin serum (Wilson Lab.) with the gastrin kit method (G-K, CIS). The former method was found to have a measurable gastrin concentration range between 60 and 1,000 pg/ml, whereas the range of the latter method was between 25 and 800 pg/ml. The reproducibility of each method was satisfactory. The G-K method was affected more readily by co-existing proteins, whereas the interferences by other biologically active factors, e.g., CCK/PZ, caerulein, etc., were negligible. While there was a highly significant correlation between the values, values obtained by the G-K method were generally slightly lower than the values obtained by the double antibody method. Results of fractionation analysis employing gel filtration of blood and tissue immunoreactive gastrin caused the authors to observe that the value of big gastrin as determined with the G-K method was lower than that obtained by the double antibody method, and that the difference was especially remarkable for gastrin in blood.

  14. AKTIVITAS KOMBINASI EKSTRAK ETANOL DAUN SIRSAK (Annona muricata L. DAN DAUN PEGAGAN (Centella asiatica L.Urb TERHADAP KELARUTAN KALSIUM BATU GINJAL SECARA IN VITRO

    Directory of Open Access Journals (Sweden)

    Ni Wayan Swintari

    2017-03-01

    Full Text Available Soursop (Annona muricata L. and gotu kola (Centella asiatica L.Urb is a plant that can be used as a laxative medicine kidney stones. This is because of the content of bioactive compounds including flavonoids, especially the leaf section. The use of a combination of soursop leaf extract and gotu kola can improve their effectiveness in order to remove calcium kidney stones. Therefore, this study aims to determine the activity and to determine dose combination soursop leaf ethanol extract (SS and gotu kola leaf (GK which is effective for dissolving potassium kidney stones. Extracts prepared by maceration method using ethanol 96% then subsequently tested phytochemical screening and thin layer chromatography profiles on each extract. Results chromatogram showed the extract containing flavonoids. Testing the activity of the combination of soursop and gotu kola extract performed using seven treatments, combination 1 (SS 1% + GK 7.5% , a combination of 2 (SS  0.5% + GK 3.75%, the combination of 3 (SS 0, 25% + GK 1.875%, single soursop leaf extract (1%, single gotu kola extract (7.5%, negative control (aquadest and positive control (Batugin elixir. Kidney stones are then put as much as 100 mg in each treatment and incubated for 6 hours (37 ℃. Results filtrate then didestruksi and dissolved calcium levels measured using Atomic Absorption Spectrophotometer. The results showed the combination soursop leaf ethanol extract and gotu kola leaf extract with a combination of 2 (SS 0.5% + GK 3.75% is an effective combination of the ability to dissolve calcium kidney stones in vitro.

  15. Feasibility of identification of gamma knife planning strategies by identification of pareto optimal gamma knife plans.

    Science.gov (United States)

    Giller, C A

    2011-12-01

    The use of conformity indices to optimize Gamma Knife planning is common, but does not address important tradeoffs between dose to tumor and normal tissue. Pareto analysis has been used for this purpose in other applications, but not for Gamma Knife (GK) planning. The goal of this work is to use computer models to show that Pareto analysis may be feasible for GK planning to identify dosimetric tradeoffs. We define a GK plan A to be Pareto dominant to B if the prescription isodose volume of A covers more tumor but not more normal tissue than B, or if A covers less normal tissue but not less tumor than B. A plan is Pareto optimal if it is not dominated by any other plan. Two different Pareto optimal plans represent different tradeoffs between dose to tumor and normal tissue, because neither plan dominates the other. 'GK simulator' software calculated dose distributions for GK plans, and was called repetitively by a genetic algorithm to calculate Pareto dominant plans. Three irregular tumor shapes were tested in 17 trials using various combinations of shots. The mean number of Pareto dominant plans/trial was 59 ± 17 (sd). Different planning strategies were identified by large differences in shot positions, and 70 of the 153 coordinate plots (46%) showed differences of 5mm or more. The Pareto dominant plans dominated other nearby plans. Pareto dominant plans represent different dosimetric tradeoffs and can be systematically calculated using genetic algorithms. Automatic identification of non-intuitive planning strategies may be feasible with these methods.

  16. Skin-specific regulation of SREBP processing and lipid biosynthesis by glycerol kinase 5

    OpenAIRE

    Zhang, Duanwu; Tomisato, Wataru; Su, Lijing; Sun, Lei; Choi, Jin Huk; Zhang, Zhao; Wang, Kuan-wen; Zhan, Xiaoming; Choi, Mihwa; Li, Xiaohong; Tang, Miao; Castro-Perez, Jose M.; Hildebrand, Sara; Murray, Anne R.; Moresco, Eva Marie Y.

    2017-01-01

    We discovered a previously unrecognized regulator of cholesterol biosynthesis, glycerol kinase 5 (GK5), which functions exclusively in the skin independently of cholesterol regulation in other tissues. GK5 negatively regulates the processing and nuclear localization of sterol regulatory element binding proteins, transcription factors that control expression of virtually all cholesterol synthesis enzymes. Excessive amounts of cholesterol, triglycerides, and ceramides were found in the skin of ...

  17. Experimental and Theoretical Approaches To Investigate the Immunogenicity of Taenia solium-Derived KE7 Antigen.

    Science.gov (United States)

    Bobes, Raúl J; Navarrete-Perea, José; Ochoa-Leyva, Adrián; Anaya, Víctor Hugo; Hernández, Marisela; Cervantes-Torres, Jacquelynne; Estrada, Karel; Sánchez-Lopez, Filiberto; Soberón, Xavier; Rosas, Gabriela; Nunes, Cáris Maroni; García-Varela, Martín; Sotelo-Mundo, Rogerio Rafael; López-Zavala, Alonso Alexis; Gevorkian, Goar; Acero, Gonzalo; Laclette, Juan P; Fragoso, Gladis; Sciutto, Edda

    2017-12-01

    Taenia solium cysticercosis, a parasitic disease that affects human health in various regions of the world, is preventable by vaccination. Both the 97-amino-acid-long KETc7 peptide and its carboxyl-terminal, 18-amino-acid-long sequence (GK-1) are found in Taenia crassiceps Both peptides have proven protective capacity against cysticercosis and are part of the highly conserved, cestode-native, 264-amino-acid long protein KE7. KE7 belongs to a ubiquitously distributed family of proteins associated with membrane processes and may participate in several vital cell pathways. The aim of this study was to identify the T. solium KE7 (TsKE7) full-length protein and to determine its immunogenic properties. Recombinant TsKE7 (rTsKE7) was expressed in Escherichia coli Rosetta2 cells and used to obtain mouse polyclonal antibodies. Anti-rTsKE7 antibodies detected the expected native protein among the 350 spots developed from T. solium cyst vesicular fluid in a mass spectrometry-coupled immune proteomic analysis. These antibodies were then used to screen a phage-displayed 7-random-peptide library to map B-cell epitopes. The recognized phages displayed 9 peptides, with the consensus motif Y(F/Y)PS sequence, which includes YYYPS (named GK-1M, for being a GK-1 mimotope), exactly matching a part of GK-1. GK-1M was recognized by 58% of serum samples from cysticercotic pigs with 100% specificity but induced weak protection against murine cysticercosis. In silico analysis revealed a universal T-cell epitope(s) in native TsKE7 potentially capable of stimulating cytotoxic T lymphocytes and helper T lymphocytes under different major histocompatibility complex class I and class II mouse haplotypes. Altogether, these results provide a rationale for the efficacy of the KETc7, rTsKE7, and GK-1 peptides as vaccines. Copyright © 2017 American Society for Microbiology.

  18. VP24-Karyopherin Alpha Binding Affinities Differ between Ebolavirus Species, Influencing Interferon Inhibition and VP24 Stability.

    Science.gov (United States)

    Schwarz, Toni M; Edwards, Megan R; Diederichs, Audrey; Alinger, Joshua B; Leung, Daisy W; Amarasinghe, Gaya K; Basler, Christopher F

    2017-02-15

    Zaire ebolavirus (EBOV), Bundibugyo ebolavirus (BDBV), and Reston ebolavirus (RESTV) belong to the same genus but exhibit different virulence properties. VP24 protein, a structural protein present in all family members, blocks interferon (IFN) signaling and likely contributes to virulence. Inhibition of IFN signaling by EBOV VP24 (eVP24) involves its interaction with the NPI-1 subfamily of karyopherin alpha (KPNA) nuclear transporters. Here, we evaluated eVP24, BDBV VP24 (bVP24), and RESTV VP24 (rVP24) interactions with three NPI-1 subfamily KPNAs (KPNA1, KPNA5, and KPNA6). Using purified proteins, we demonstrated that each VP24 binds to each of the three NPI-1 KPNAs. bVP24, however, exhibited approximately 10-fold-lower KPNA binding affinity than either eVP24 or rVP24. Cell-based assays also indicate that bVP24 exhibits decreased KPNA interaction, decreased suppression of IFN induced gene expression, and a decreased half-life in transfected cells compared to eVP24 or rVP24. Amino acid sequence alignments between bVP24 and eVP24 also identified residues within and surrounding the previously defined eVP24-KPNA5 binding interface that decrease eVP24-KPNA affinity or bVP24-KPNA affinity. VP24 mutations that lead to reduced KPNA binding affinity also decrease IFN inhibition and shorten VP24 half-lives. These data identify novel functional differences in VP24-KPNA interaction and reveal a novel impact of the VP24-KPNA interaction on VP24 stability. The interaction of Ebola virus (EBOV) VP24 protein with host karyopherin alpha (KPNA) proteins blocks type I interferon (IFN) signaling, which is a central component of the host innate immune response to viral infection. Here, we quantitatively compared the interactions of VP24 proteins from EBOV and two members of the Ebolavirus genus, Bundibugyo virus (BDBV) and Reston virus (RESTV). The data reveal lower binding affinity of the BDBV VP24 (bVP24) for KPNAs and demonstrate that the interaction with KPNA modulates inhibition

  19. Temperature Dependence of the Stability of Ion Pair Interactions ...

    Indian Academy of Sciences (India)

    The occurrence of bridging water molecules between the ions ensures that the ions are not ... The structural features that render this thermostability ..... dehydrogenase single site mutant T198I from Thermus thermophilus with PDB ID 1BDM.

  20. VP24-Karyopherin Alpha Binding Affinities Differ between Ebolavirus Species, Influencing Interferon Inhibition and VP24 Stability

    Energy Technology Data Exchange (ETDEWEB)

    Schwarz, Toni M.; Edwards, Megan R.; Diederichs, Audrey; Alinger, Joshua B.; Leung, Daisy W.; Amarasinghe, Gaya K.; Basler, Christopher F.; Lyles, Douglas S.

    2016-12-14

    ABSTRACT

    Zaire ebolavirus(EBOV),Bundibugyo ebolavirus(BDBV), andReston ebolavirus(RESTV) belong to the same genus but exhibit different virulence properties. VP24 protein, a structural protein present in all family members, blocks interferon (IFN) signaling and likely contributes to virulence. Inhibition of IFN signaling by EBOV VP24 (eVP24) involves its interaction with the NPI-1 subfamily of karyopherin alpha (KPNA) nuclear transporters. Here, we evaluated eVP24, BDBV VP24 (bVP24), and RESTV VP24 (rVP24) interactions with three NPI-1 subfamily KPNAs (KPNA1, KPNA5, and KPNA6). Using purified proteins, we demonstrated that each VP24 binds to each of the three NPI-1 KPNAs. bVP24, however, exhibited approximately 10-fold-lower KPNA binding affinity than either eVP24 or rVP24. Cell-based assays also indicate that bVP24 exhibits decreased KPNA interaction, decreased suppression of IFN induced gene expression, and a decreased half-life in transfected cells compared to eVP24 or rVP24. Amino acid sequence alignments between bVP24 and eVP24 also identified residues within and surrounding the previously defined eVP24-KPNA5 binding interface that decrease eVP24-KPNA affinity or bVP24-KPNA affinity. VP24 mutations that lead to reduced KPNA binding affinity also decrease IFN inhibition and shorten VP24 half-lives. These data identify novel functional differences in VP24-KPNA interaction and reveal a novel impact of the VP24-KPNA interaction on VP24 stability.

    IMPORTANCEThe interaction of Ebola virus (EBOV) VP24 protein with host karyopherin alpha (KPNA) proteins blocks type I interferon (IFN) signaling, which is a central component of the host innate immune response to viral infection. Here, we quantitatively compared the

  1. Decay of a Jπ=36+ Resonance in the 24Mg+24Mg Reaction

    International Nuclear Information System (INIS)

    Salsac, M.-D.; Haas, F.; Courtin, S.

    2005-01-01

    For the 24 Mg+ 24 Mg reaction, striking narrow and correlated resonance structures have been observed previously in the excitation functions of the elastic and low-lying channels. In our study we have decided to focus on the resonance at E C M =45.7 MeV, which is known to have J π =36 + . Despite the very high excitation energy(∼60 MeV) in the 48 Cr composite system, this resonance has a narrow total width of 170 keV. To determine precisely which states in the inelastic 24 Mg channels carry away the resonance flux, an experiment, on the 24 Mg + 24 Mg reaction at energies On and OFF resonance, has been performed at the Legnaro Tandem accelerator using the Prisma fragment spectrometer associated with the CLARA γ array

  2. CarD uses a minor groove wedge mechanism to stabilize the RNA polymerase open promoter complex.

    Science.gov (United States)

    Bae, Brian; Chen, James; Davis, Elizabeth; Leon, Katherine; Darst, Seth A; Campbell, Elizabeth A

    2015-09-08

    A key point to regulate gene expression is at transcription initiation, and activators play a major role. CarD, an essential activator in Mycobacterium tuberculosis, is found in many bacteria, including Thermus species, but absent in Escherichia coli. To delineate the molecular mechanism of CarD, we determined crystal structures of Thermus transcription initiation complexes containing CarD. The structures show CarD interacts with the unique DNA topology presented by the upstream double-stranded/single-stranded DNA junction of the transcription bubble. We confirm that our structures correspond to functional activation complexes, and extend our understanding of the role of a conserved CarD Trp residue that serves as a minor groove wedge, preventing collapse of the transcription bubble to stabilize the transcription initiation complex. Unlike E. coli RNAP, many bacterial RNAPs form unstable promoter complexes, explaining the need for CarD.

  3. A cytocidal tissue kallikrein isolated from mouse submandibular glands.

    Science.gov (United States)

    Murakami, K; Ikigai, H; Nagumo, N; Tomita, M; Shimamura, T

    1989-11-06

    A cytocidal factor against mouse thymocytes was purified from the submandibular glands of female BALB/c mice using Sephadex G-50 gel filtration chromatography and reverse-phase HPLC. SDS-PAGE and amino acid sequence analysis revealed that the cytocidal factor was mouse glandular kallikrein (mGK)-6. mGK-6 showed an optimal enzyme activity at pH 10 and a cytocidal activity against thymocytes in a dose-dependent manner.

  4. Insulin-induced activation of glycerol-3-phosphate acyltransferase by a chiro-inositol-containing insulin mediator is defective in adipocytes of insulin-resistant, type II diabetic, Goto-Kakizaki rats.

    OpenAIRE

    Farese, R V; Standaert, M L; Yamada, K; Huang, L C; Zhang, C; Cooper, D R; Wang, Z; Yang, Y; Suzuki, S; Toyota, T

    1994-01-01

    Type II diabetic Goto-Kakizaki (GK) rats were insulin-resistant in euglycemic-hyperinsulinemic clamp studies. We therefore examined insulin signaling systems in control Wistar and diabetic GK rats. Glycerol-3-phosphate acyltransferase (G3PAT), which is activated by headgroup mediators released from glycosyl-phosphatidylinositol (GPI), was activated by insulin in intact and cell-free adipocyte preparations of control, but not diabetic, rats. A specific chiro-inositol-containing inositol phosph...

  5. Characterization of Thermostable Cellulases Produced by Bacillus and Geobacillus Strains

    Science.gov (United States)

    Bacterial community composition of thermophilic (60 deg C) mixed cellulose-enrichment cultures was examined by constructing a 16S rDNA clone library which demonstrated major lineages affiliated to Actinobacteria, Bacteroidetes, Chloroflexi, Deinococcus-Thermus, Firmicutes, and Proteobacteria. A tot...

  6. Epigenetic Regulation of Vitamin D 24-Hydroxylase/CYP24A1 in Human Prostate Cancer

    Science.gov (United States)

    Luo, Wei; Karpf, Adam R.; Deeb, Kristin K.; Muindi, Josephia R.; Morrison, Carl D.; Johnson, Candace S.; Trump, Donald L.

    2010-01-01

    Calcitriol, a regulator of calcium homeostasis with antitumor properties, is degraded by the product of the CYP24A1 gene which is downregulated in human prostate cancer by unknown mechanisms. We found that CYP24A1 expression is inversely correlated with promoter DNA methylation in prostate cancer cell lines. Treatment with the DNA methyltransferase inhibitor 5-aza-2′-deoxycytidine (DAC) activates CYP24A1 expression in prostate cancer cells. In vitro methylation of the CYP24A1 promoter represses its promoter activity. Furthermore, inhibition of histone deacetylases by trichostatin A (TSA) enhances the expression of CYP24A1 in prostate cancer cells. ChIP-qPCR reveals that specific histone modifications are associated with the CYP24A1 promoter region. Treatment with TSA increases H3K9ac and H3K4me2 and simultaneously decreases H3K9me2 at the CYP24A1 promoter. ChIP-qPCR assay reveals that treatment with DAC and TSA increases the recruitment of VDR to the CYP24A1 promoter. RT-PCR analysis of paired human prostate samples reveals that CYP24A1 expression is down-regulated in prostate malignant lesions compared to adjacent histologically benign lesions. Bisulfite pyrosequencing shows that CYP24A1 gene is hypermethylated in malignant lesions compared to matched benign lesions. Our findings indicate that repression of CYP24A1 gene expression in human prostate cancer cells is mediated in part by promoter DNA methylation and repressive histone modifications. PMID:20587525

  7. Progress of interleukin-24 study

    International Nuclear Information System (INIS)

    Liu Yongzhe; Jin Shunzi

    2007-01-01

    Interteukin-24 (11-24)is a member of interleukin-10 family. Besides its character as a cytokine, IL-24 could induce apoptosis of different kinds of tumor cells without any harmful effects to normal cells. IL-24 could both promote bystander anti-tumor effect, inhibit tumor growth and angiogenesis in animal models. It also has effects of radiosensitization and immunoregulation. (authors)

  8. Signaling through cGMP-dependent protein kinase I in the amygdala is critical for auditory-cued fear memory and long-term potentiation.

    Science.gov (United States)

    Paul, Cindy; Schöberl, Florian; Weinmeister, Pascal; Micale, Vincenzo; Wotjak, Carsten T; Hofmann, Franz; Kleppisch, Thomas

    2008-12-24

    Long-term potentiation (LTP) of inputs relaying sensory information from cortical and thalamic neurons to principal neurons in the lateral amygdala (LA) is thought to serve as a cellular mechanism for associative fear learning. Nitric oxide (NO), a messenger molecule widely implicated in synaptic plasticity and behavior, has been shown to enhance LTP in the LA as well as consolidation of associative fear memory. Additional evidence suggests that NO-induced enhancement of LTP and amygdala-dependent learning requires signaling through soluble guanylyl cyclase (sGC) and cGMP-dependent protein kinase (cGK). Mammals possess two genes for cGK: the prkg1 gene gives rise to the cGK type I isoforms, cGKIalpha and cGKIbeta, and the prkg2 gene encodes the cGK type II. Reportedly, both cGKI and cGKII are expressed in the amygdala, and cGKII is involved in controlling anxiety-like behavior. Because selective pharmacological tools for individual cGK isoforms are lacking, we used different knock-out mouse models to examine the function of cGKI and cGKII for LTP in the LA and pavlovian fear conditioning. We found robust expression of the cGKI specifically in the LA with cGKIbeta as the prevailing isoform. We further show a marked reduction of LTP at both thalamic and cortical inputs to the LA and a selective impairment of auditory-cued fear memory in cGKI-deficient mutants. In contrast, cGKII null mutants lack these phenotypes. Our data suggest a function of cGKI, likely the beta isoform, in the LA, supporting synaptic plasticity and consolidation of fear memory.

  9. Tri-trophic interactions between Bt cotton, the herbivore Aphis gossypii Glover (Homoptera: Aphididae), and the predator Chrysopa pallens (Rambur) (Neuroptera: Chrysopidae).

    Science.gov (United States)

    Guo, Jian-Ying; Wan, Fang-Hao; Dong, Liang; Lövei, Gábor L; Han, Zhao-Jun

    2008-02-01

    Tri-trophic impacts of transgenic Bacillus thuringiensis (Bt) cotton GK12 and NuCOTN 99B were studied using a predator, the great lacewing Chrysopa pallens (Rambur), and its prey, the cotton aphid Aphis gossypii Glover, in laboratory feeding experiments. The parental nontransgenic cotton cultivar of GK12 was used as control. The predator was fed with uniform (aphids from a single cultivar) or mixed prey (aphids from the three cotton cultivars provided on alternate days). Mortality and development of the immature stages, pupal body mass, adult sex ratio, fecundity, and egg viability of C. pallens were measured. When fed GK12-originated aphid prey, pupal body mass of C. pallens was significantly higher than that of the control, more females emerged, and these females laid significantly more eggs. Other parameters were not impacted. Females emerging from larvae maintained on NuCOTN 99B-originated prey laid fewer eggs than those maintained on GK12. Other measurements did not differ significantly between the two Bt cotton cultivars. Compared with the control, mixed feeding significantly prolonged pupal development time and increased pupal body mass and percentage of females but did not affect other parameters. These results indicate that C. pallens is sensitive to aphid prey from different cotton cultivars. Transgenic Bt cotton GK12-originated aphid prey has no adverse impact on survival, development, and fecundity of C. pallens. Between the two Bt cotton cultivars, NuCOTN 99B-originated aphid prey provided to C. pallens in the larval stage may lower female fecundity. Mixed feeding of C. pallens with the two Bt cotton-originated prey and non-Bt prey may have some adverse impacts on pupal development.

  10. The methanol seed extract of Garcinia kola attenuated angiotensin II- and lipopolyssacharide-inducedvascular smooth muscle cell proliferation and nitric oxide production

    Directory of Open Access Journals (Sweden)

    Adeolu A. Adedapo

    2016-10-01

    Full Text Available All over the world, cardiovascular diseases are a risk factor for poor health and early death with predisposing factors to include age, gender, tobacco use, physical inactivity, excessive alcohol consumption, unhealthy diet, obesity, family history of cardiovascular disease, hypertension, diabetes mellitus, hyperlipidemia, psychosocial factors, poverty and low educational status, and air pollution. It is envisaged that herbal products that can stem this trend would be of great benefit. Garcinia kola (GK, also known as bitter kola is one of such plants. Generally used as a social snack and offered to guests in some cultural settings, bitter kola has been indicated in the treatment of laryngitis, general inflammation, bronchitis, viral infections and diabetes. In this study, the effects of methanol seed extract of Garcinia kola on the proliferation of Vascular Smooth Muscle Cells (VSMCs in cell culture by Angiotensin II (Ang II and LPS-induced NO production were carried out. Confluent VSMCs were exposed to GK (25, 50 and 100 μg/ml before or after treatment with lipopolyssacharide (100μg/ml, and Angiotensin II (10-8-10-6M. Cellular proliferation was determined by MTT assay and NO production by Griess assay. Treatment with Angiotensin II (10-8, 10-6 or LPS significantly enhanced proliferation of VSM cells while LPS significantly increased nitric oxide (NO production. Treatment with GK (25, 50 & 100 μg/ml attenuated VSM cell proliferation. The results indicate that GK has potential to inhibit mitogen activated vascular cell growth and possibly inhibit inflammatory responses to LPS. Thus GK may be useful in condition that is characterized by cellular proliferation and inflammatory responses.

  11. A new spectroscopic calibration to determine Teff and [Fe/H] of FGK dwarfs and giants

    Directory of Open Access Journals (Sweden)

    Teixeira G. D. C.

    2017-01-01

    Full Text Available We present a new spectroscopic calibration for a fast estimate of Teff and [Fe/H] for FGK dwarfs and GK giant stars. We used spectra from a joint sample of 708 stars, composed by 451 FGK dwarfs and 257 GK-giant stars with homogeneously determined spectroscopic stellar parameters. We have derived 322 EW line-ratios and 100 FeI lines that can be used to compute Teff and [Fe/H], respectively. We show that these calibrations are effective for FGK dwarfs and GK-giant stars in the following ranges: 4500 K < Teff < 6500 K, 2.5 < log g < 4.9 dex, and –0.8 < [Fe/H] < 0:5 dex. The new calibration has a standard deviation of 74 K for Teff and 0.07 dex for [Fe/H]. We use four independent samples of stars to test and verify the new calibration, a sample of giant stars, a sample composed of Gaia FGK benchmark stars, a sample of GK-giant stars from the DR1 of the Gaia-ESO survey, and a sample of FGK-dwarf stars. We present a new computer code, GeTCal, for automatically producing new calibration files based on any new sample of stars.

  12. Fusion potentials for Gk and handle squashing

    International Nuclear Information System (INIS)

    Crescimanno, M.

    1993-01-01

    Using Chern-Simons gauge theory, we show that the fusion ring of the conformal field theory G k (G any Lie algebra) is isomorphic to P[u]/(∇V) where (∇V) is the ideal generated by conditions ∇V=0. We explicitly construct V for all G k . We also derive a residue-like formula for the correlation functions in the Chern-Simons theory thus providing an RCFT version of the residue formula for the topological Landau-Ginzburg model. An operator that acts like a measure in this residue formula has the interpretation of a handle-squashing operator and explicit formulae for this operator are given. (orig.)

  13. 24. International Conference on Atomic Collisions in Solids ICACS-24

    International Nuclear Information System (INIS)

    2010-01-01

    This Book contains the abstracts of invited and contributed talks submitted for presentation at the 24 th International Conference on Atomic Collisions in Solids - ICACS-24. Out of nearly 200 submitted abstracts the International Programme Committee selected 46 oral and 89 poster contributions. Furthermore, 15 plenary invited lectures and the honorary Lindhard lecture are included in the scientific program. An additional tutorial day with 4 tutorial lectures is organised on Sunday prior to the Conference.

  14. Diet-dependent depletion of queuosine in tRNAs in Caenorhabditis ...

    Indian Academy of Sciences (India)

    PRAKASH KUMAR

    1972). Apart from the known exceptions of yeast, Thermus .... Hypomodification can also be induced by .... Panels C and D show UV absorbance of the Q and manQ peaks, respectively. ... E. coli B105(Q–), a mutant strain lacking Q modification.

  15. Development of an immunoassay for determination of 2,4-dichlorophenoxyacetic acid (2,4-D) based upon the recombinant Fab fragment of 2,4-D specific antibody

    Science.gov (United States)

    Nguyen, Van C.; Nguyen, Thi D. T.; Dau, Hung A.; Tham, Thu N.; Quyen, Dinh T.; Bachmman, Till; Schmid, Rolf D.

    2001-09-01

    To develop an immunoassay and further an immunosensor for 2,4-D based upon recombinant antibody, the Fab fragments of 2,4-D specific antibody were expressed in E. coli. Western blotting analysis of the periplasmic cell fractions shown that under the non-reducing condition only a single protein band at a molecular mass of 45-kDa, corresponding to the whole Fab fragment was detected. Antigen binding activity for 2,4-D was found only in the extract of cells bearing the 2,4-D plasmid. An immunoassay based on the competitive reaction of 2,4-D and enzyme tracer with 2,4-D Fab fragments immobilized on micro titer plates via rabbit anti-mouse IgC was developed. Using this assay, 2,4-D could be detected at concentration range of 0.5 (mu) g/1 to 10(mu) g/1. The center point of the 2,4-D test was found at a concentration of 5 (mu) g/l. The assay was applied for detection of 2,4-D in spiked orange samples, resulting in recovery rate of 90 percent. The immunoassay could be applied to monitor human exposure to 2,4-D from contamination in fruit samples.

  16. 7 CFR 1230.24 - State.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 10 2010-01-01 2010-01-01 false State. 1230.24 Section 1230.24 Agriculture... CONSUMER INFORMATION Pork Promotion, Research, and Consumer Information Order Definitions § 1230.24 State. State means each of the 50 States. ...

  17. Disparate effects of p24alpha and p24delta on secretory protein transport and processing.

    Directory of Open Access Journals (Sweden)

    Jeroen R P M Strating

    Full Text Available BACKGROUND: The p24 family is thought to be somehow involved in endoplasmic reticulum (ER-to-Golgi protein transport. A subset of the p24 proteins (p24alpha(3, -beta(1, -gamma(3 and -delta(2 is upregulated when Xenopus laevis intermediate pituitary melanotrope cells are physiologically activated to produce vast amounts of their major secretory cargo, the prohormone proopiomelanocortin (POMC. METHODOLOGY/PRINCIPAL FINDINGS: Here we find that transgene expression of p24alpha(3 or p24delta(2 specifically in the Xenopus melanotrope cells in both cases causes an effective displacement of the endogenous p24 proteins, resulting in severely distorted p24 systems and disparate melanotrope cell phenotypes. Transgene expression of p24alpha(3 greatly reduces POMC transport and leads to accumulation of the prohormone in large, ER-localized electron-dense structures, whereas p24delta(2-transgenesis does not influence the overall ultrastructure of the cells nor POMC transport and cleavage, but affects the Golgi-based processes of POMC glycomaturation and sulfation. CONCLUSIONS/SIGNIFICANCE: Transgenic expression of two distinct p24 family members has disparate effects on secretory pathway functioning, illustrating the specificity and non-redundancy of our transgenic approach. We conclude that members of the p24 family furnish subcompartments of the secretory pathway with specific sets of machinery cargo to provide the proper microenvironments for efficient and correct secretory protein transport and processing.

  18. 48 CFR 24.201 - Authority.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 1 2010-10-01 2010-10-01 false Authority. 24.201 Section 24.201 Federal Acquisition Regulations System FEDERAL ACQUISITION REGULATION SOCIOECONOMIC PROGRAMS PROTECTION OF PRIVACY AND FREEDOM OF INFORMATION Freedom of Information Act 24.201 Authority. The Freedom of...

  19. 28 CFR 24.304 - Settlement.

    Science.gov (United States)

    2010-07-01

    ... 28 Judicial Administration 1 2010-07-01 2010-07-01 false Settlement. 24.304 Section 24.304... DEPARTMENT OF JUSTICE ADMINISTRATIVE PROCEEDINGS Procedures for Considering Applications § 24.304 Settlement. A prevailing party and Department counsel may agree on a proposed settlement of an award before...

  20. 7 CFR 2201.24 - Insurance.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 15 2010-01-01 2010-01-01 false Insurance. 2201.24 Section 2201.24 Agriculture Regulations of the Department of Agriculture (Continued) LOCAL TELEVISION LOAN GUARANTEE BOARD LOCAL TELEVISION LOAN GUARANTEE PROGRAM-PROGRAM REGULATIONS Loan Guarantees § 2201.24 Insurance. The Borrower of a...

  1. 49 CFR 24.108 - Donations.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 1 2010-10-01 2010-10-01 false Donations. 24.108 Section 24.108 Transportation Office of the Secretary of Transportation UNIFORM RELOCATION ASSISTANCE AND REAL PROPERTY ACQUISITION FOR FEDERAL AND FEDERALLY-ASSISTED PROGRAMS Real Property Acquisition § 24.108 Donations. An owner whose real...

  2. 40 CFR 17.24 - Settlement.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 1 2010-07-01 2010-07-01 false Settlement. 17.24 Section 17.24... JUSTICE ACT IN EPA ADMINISTRATIVE PROCEEDINGS Procedures for Considering Applications § 17.24 Settlement. A prevailing party and EPA counsel may agree on a proposed settlement of an award before final...

  3. 12 CFR Appendix 1 to Part 24 - CD-1-National Bank Community Development (Part 24) Investments

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 1 2010-01-01 2010-01-01 false CD-1-National Bank Community Development (Part 24) Investments 1 Appendix 1 To Part 24 Banks and Banking COMPTROLLER OF THE CURRENCY, DEPARTMENT OF... PUBLIC WELFARE INVESTMENTS Pt. 24, App. 1 Appendix 1 To Part 24—CD-1—National Bank Community Development...

  4. Radioimmunoassay for 2,4-dichlorophenoxyacetic acid

    International Nuclear Information System (INIS)

    Knopp, D.; Dobberkau, H.J.; Nuhn, P.

    1985-01-01

    Antisera to 2,4-dichlorophenoxyacetic acid (2,4-D), a widely used herbicide, have been obtained from rabbits following immunization with various 2,4-D-protein conjugates. Employing [ 125 I] 2,4-D-tyramine as the radioligand for the antisera, very poor assay sensitivity was achieved because of a much higher affinity of the antibodies to the tracer. When using [6- 3 H] 2,4-D (specific radioactivity 465 GBq/mmol) a sensitive and specific radioimmunoassay (RIA) for 2,4-D could be developed, which allows determination directly in water, plasma and urine samples. Levels as low as approximately 100 pg (450 femtomoles) of 2,4-D can be detected. The antiserum is fairly specific for 2,4-D. Other related phenoxycarboxylic acids and dichlorophenol showed a cross-reactivity smaller than 10%. After a single administration of 2,4-D (0.91 mg/100 g body weight, orally) to rats, plasma and urine levels were determined at different times. Results correspond to those found in the literature, thus indicating the utility of the RIA. Further applications and limitations are discussed. (orig.)

  5. 43 CFR 24.1 - Introduction.

    Science.gov (United States)

    2010-10-01

    ... 43 Public Lands: Interior 1 2010-10-01 2010-10-01 false Introduction. 24.1 Section 24.1 Public Lands: Interior Office of the Secretary of the Interior DEPARTMENT OF THE INTERIOR FISH AND WILDLIFE POLICY: STATE-FEDERAL RELATIONSHIPS § 24.1 Introduction. (a) In 1970, the Secretary of the Interior...

  6. 10 CFR 725.24 - Administration.

    Science.gov (United States)

    2010-01-01

    ... 10 Energy 4 2010-01-01 2010-01-01 false Administration. 725.24 Section 725.24 Energy DEPARTMENT OF ENERGY PERMITS FOR ACCESS TO RESTRICTED DATA Permits § 725.24 Administration. With respect to each permit... designate a DOE or National Nuclear Security Administration office which will: (a) Process all personnel...

  7. 19 CFR 207.24 - Hearing.

    Science.gov (United States)

    2010-04-01

    ... 19 Customs Duties 3 2010-04-01 2010-04-01 false Hearing. 207.24 Section 207.24 Customs Duties... EXPORTS TO THE UNITED STATES Final Determinations, Short Life Cycle Products § 207.24 Hearing. (a) In general. The Commission shall hold a hearing concerning an investigation before making a final...

  8. Equine herpesvirus type 1 (EHV-1) glycoprotein K is required for efficient cell-to-cell spread and virus egress

    International Nuclear Information System (INIS)

    Neubauer, Antonie; Osterrieder, Nikolaus

    2004-01-01

    The function of the equine herpesvirus type 1 (EHV-1) glycoprotein K (gK) homologue was investigated. Deletion of 88% of the UL53-homologous open reading frame in EHV-1 strain RacH resulted in a severe growth defect of the gK-negative virus (HΔgK) as reflected by a significant decrease in the production of infectious virus progeny on RK13 cells. The HΔgK virus induced only minute plaques, was unable to form syncytia, and its penetration efficiency into RK13 cells was reduced by approximately 40%. To further analyze gK function and intracellular trafficking, gK of strain RacH was replaced by a C-terminally truncated gK-green fluorescent protein fusion protein (gK-GFP). The generated recombinant virus was shown to replicate well on non-complementing cells, and virus penetration and syncytium formation were comparable to parental RacH. A reduction in plaque size and slightly decreased intra- and extracellular virus titers, however, were observed. The gK-GFP fusion protein was expressed with early-late kinetics, and multiple forms of the protein exhibiting M r s between 50,000 and 85,000 were detected by Western blot analysis. The various gK-GFP forms were shown to be N-glycosylated, associated with membranes of the Golgi apparatus, and were incorporated into extracellular virions. Complete processing of gK-GFP was only observed within the context of viral infection. From the results, we concluded that EHV-1 gK is required for efficient virus growth in vitro and that the carboxy-terminal amino acids are not required for its function, because the gK-GFP fusion protein was able to complement for EHV-1 growth in the absence of authentic gK

  9. 43 CFR 24.7 - Exemptions.

    Science.gov (United States)

    2010-10-01

    ... 43 Public Lands: Interior 1 2010-10-01 2010-10-01 false Exemptions. 24.7 Section 24.7 Public Lands...-FEDERAL RELATIONSHIPS § 24.7 Exemptions. (a) Exempted from this policy are the following: (1) The control and regulation by the United States, in the area in which an international convention or treaty...

  10. 19 CFR 212.24 - Settlement.

    Science.gov (United States)

    2010-04-01

    ... 19 Customs Duties 3 2010-04-01 2010-04-01 false Settlement. 212.24 Section 212.24 Customs Duties... IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT Procedures for Considering Applications § 212.24 Settlement. The applicant and the Commission may agree on a proposed settlement of the award before final action...

  11. 45 CFR 13.24 - Settlements.

    Science.gov (United States)

    2010-10-01

    ... 45 Public Welfare 1 2010-10-01 2010-10-01 false Settlements. 13.24 Section 13.24 Public Welfare... ACT IN AGENCY PROCEEDINGS Procedures for Considering Applications § 13.24 Settlements. The applicant and the agency's litigating party may agree on a proposed settlement of the award at any time prior to...

  12. 42 CFR 24.2 - Allocation.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 1 2010-10-01 2010-10-01 false Allocation. 24.2 Section 24.2 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES PERSONNEL SENIOR BIOMEDICAL RESEARCH SERVICE § 24... talent. (e) The Secretary will ensure that SBRS slots are used in support of high priority programs...

  13. 24 CFR 208.104 - Applicability.

    Science.gov (United States)

    2010-04-01

    ... Projects (24 CFR part 884); Loans for Housing for the Elderly or Handicapped (24 CFR part 885) and section...(d)(3) Below Market Interest Rate Housing for Low and Moderate Income Mortgage Insurance program (24...); (3) The section 221(d)(3) Below Market Interest Rate Housing for Low and Moderate Income Mortgage...

  14. 32 CFR 34.24 - Supplies.

    Science.gov (United States)

    2010-07-01

    ... shall retain any unused supplies. If the inventory of unused supplies exceeds $5,000 in total aggregate... 32 National Defense 1 2010-07-01 2010-07-01 false Supplies. 34.24 Section 34.24 National Defense... § 34.24 Supplies. (a) Title shall vest in the recipient upon acquisition for supplies acquired with...

  15. 15 CFR 24.33 - Supplies.

    Science.gov (United States)

    2010-01-01

    ... residual inventory of unused supplies exceeding $5,000 in total aggregate fair market value upon... 15 Commerce and Foreign Trade 1 2010-01-01 2010-01-01 false Supplies. 24.33 Section 24.33 Commerce..., and Subawards § 24.33 Supplies. (a) Title. Title to supplies acquired under a grant or subgrant will...

  16. 34 CFR 303.24 - State.

    Science.gov (United States)

    2010-07-01

    ... 34 Education 2 2010-07-01 2010-07-01 false State. 303.24 Section 303.24 Education Regulations of... General Purpose, Eligibility, and Other General Provisions § 303.24 State. Except as provided in § 303.200(b)(3), State means each of the 50 States, the Commonwealth of Puerto Rico, the District of Columbia...

  17. 27 CFR 24.82 - Samples.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Samples. 24.82 Section 24.82 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE... vinegar and salted wine as defined in § 24.215, the proprietor shall submit under separate cover at the...

  18. Biokinetic Analysis and Metabolic Fate of 2,4-D in 2,4-D-Resistant Soybean (Glycine max).

    Science.gov (United States)

    Skelton, Joshua J; Simpson, David M; Peterson, Mark A; Riechers, Dean E

    2017-07-26

    The Enlist weed control system allows the use of 2,4-D in soybean but slight necrosis in treated leaves may be observed in the field. The objectives of this research were to measure and compare uptake, translocation, and metabolism of 2,4-D in Enlist (E, resistant) and non-AAD-12 transformed (NT, sensitive) soybeans. The adjuvant from the Enlist Duo herbicide formulation (ADJ) increased 2,4-D uptake (36%) and displayed the fastest rate of uptake (U 50 = 0.2 h) among treatments. E soybean demonstrated a faster rate of 2,4-D metabolism (M 50 = 0.2 h) compared to NT soybean, but glyphosate did not affect 2,4-D metabolism. Metabolites of 2,4-D in E soybean were qualitatively different than NT. Applying 2,4-D-ethylhexyl ester instead of 2,4-D choline (a quaternary ammonium salt) eliminated visual injury to E soybean, likely due to the time required for initial de-esterification and bioactivation. Excessive 2,4-D acid concentrations in E soybean resulting from ADJ-increased uptake may significantly contribute to foliar injury.

  19. 12 CFR 1705.24 - Settlement.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 7 2010-01-01 2010-01-01 false Settlement. 1705.24 Section 1705.24 Banks and... Consideration of the Application for Award § 1705.24 Settlement. The applicant and agency counsel may agree on a proposed settlement of an award before the final decision on the application for award is made, either in...

  20. Comparative Study on Different Expression Hosts for Alkaline Phytase Engineered in Escherichia coli.

    Science.gov (United States)

    Chen, Weiwei; Yu, Hongwei; Ye, Lidan

    2016-07-01

    The application of alkaline phytase as a feed additive is restricted by the poor specific activity. Escherichia coli is a frequently used host for directed evolution of proteins including alkaline phytase towards improved activity. However, it is not suitable for production of food-grade products due to potential pathogenicity. To combine the advantages of different expression systems, mutants of the alkaline phytase originated from Bacillus subtilis 168 (phy168) were first generated via directed evolution in E. coli and then transformed to food-grade hosts B. subtilis and Pichia pastoris for secretory expression. In order to investigate the suitability of different expression systems, the phy168 mutants expressed in different hosts were characterized and compared in terms of specific activity, pH profile, pH stability, temperature profile, and thermostability. The specific activity of B. subtilis-expressed D24G/K70R/K111E/N121S mutant at pH 7.0 and 60 °C was 30.4 U/mg, obviously higher than those in P. pastoris (22.7 U/mg) and E. coli (19.7 U/mg). Moreover, after 10 min incubation at 80 °C, the B. subtilis-expressed D24G/K70R/K111E/N121S retained about 70 % of the activity at pH 7.0 and 37 °C, whereas the values were only about 25 and 50 % when expressed in P. pastoris and E. coli, respectively. These results suggested B. subtilis as an appropriate host for expression of phy168 mutants and that the strategy of creating mutants in one host and expressing them in another might be a new solution to industrial production of proteins with desired properties.

  1. Mineralization of 2,4-Dichlorophenoxyacetic Acid (2,4-D) and Mixtures of 2,4-D and 2,4,5-Trichlorophenoxyacetic Acid by Phanerochaete chrysosporium

    Science.gov (United States)

    Yadav, J. S.; Reddy, C. A.

    1993-01-01

    Evidence is presented for mineralization of 2,4-dichlorophenoxyacetic acid (2,4-D) in nutrient-rich media (high-nitrogen and malt extract media) by wild-type Phanerochaete chrysosporium and by a peroxidase-negative mutant of this organism. Mass balance analysis of [U-ring-14C]2,4-D mineralization in malt extract cultures showed 82.7% recovery of radioactivity. Of this, 38.6% was released as 14CO2 and 27.0, 11.2, and 5.9% were present in the aqueous, methylene chloride, and mycelial fractions, respectively. 2,4-D and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) were simultaneously mineralized when presented as a mixture, and mutual inhibition of degradation was not observed. In contrast, a relatively higher rate of mineralization of 2,4-D and 2,4,5-T was observed when these compounds were tested as mixtures than when they were tested alone. PMID:16349039

  2. Kinetic analysis of human CYP24A1 metabolism of vitamin D via the C24-oxidation pathway.

    Science.gov (United States)

    Tieu, Elaine W; Tang, Edith K Y; Tuckey, Robert C

    2014-07-01

    CYP24A1 is the multicatalytic cytochrome P450 responsible for the catabolism of vitamin D via the C23- and C24-oxidation pathways. We successfully expressed the labile human enzyme in Escherichia coli and partially purified it in an active state that permitted detailed characterization of its metabolism of 1,25-dihydroxyvitamin D3 [1,25(OH)2 D3] and the intermediates of the C24-oxidation pathway in a phospholipid-vesicle reconstituted system. The C24-oxidation pathway intermediates, 1,24,25-trihydroxyvitamin D3, 24-oxo-1,25-dihydroxyvitamin D3, 24-oxo-1,23,25-trihydroxyvitamin D3 and tetranor-1,23-dihydroxyvitamin D3, were enzymatically produced from 1,25(OH)2 D3 using rat CYP24A1. Both 1,25(OH)2 D3 and 1,23-dihydroxy-24,25,26,27-tetranorvitamin D3 were found to partition strongly into the phospholipid bilayer when in aqueous medium. Changes to the phospholipid concentration did not affect the kinetic parameters for the metabolism of 1,25(OH)2 D3 by CYP24A1, indicating that it is the concentration of substrates in the membrane phase (mol substrate·mol phospholipid(-1) ) that determines their rate of metabolism. CYP24A1 exhibited Km values for the different C24-intermediates ranging from 0.34 to 15 mmol·mol phospholipid(-1) , with 24-oxo-1,23,25-trihydroxyvitamin D3 [24-oxo-1,23,25(OH)3 D3] displaying the lowest and 1,24,25-trihydroxyvitamin D3 [1,24,25(OH)3 D3] displaying the highest. The kcat values varied by up to 3.8-fold, with 1,24,25(OH)3 D3 displaying the highest kcat (34 min(-1) ) and 24-oxo-1,23,25(OH)3 D3 the lowest. The data show that the cleavage of the side chain of 24-oxo-1,23,25(OH)3 D3 occurs with the highest catalytic efficiency (kcat /Km ) and produces 1-hydroxy-23-oxo-24,25,26,27-tetranorvitamin D3 and not 1,23-dihydroxy-24,25,26,27-tetranorvitamin D3, as the primary product. These kinetic analyses also show that intermediates of the C24-oxidation pathway effectively compete with precursor substrates for binding to the active site of the

  3. Gluconeogenesis in Leishmania mexicana

    Science.gov (United States)

    Rodriguez-Contreras, Dayana; Hamilton, Nicklas

    2014-01-01

    Gluconeogenesis is an active pathway in Leishmania amastigotes and is essential for their survival within the mammalian cells. However, our knowledge about this pathway in trypanosomatids is very limited. We investigated the role of glycerol kinase (GK), phosphoenolpyruvate carboxykinase (PEPCK), and pyruvate phosphate dikinase (PPDK) in gluconeogenesis by generating the respective Leishmania mexicana Δgk, Δpepck, and Δppdk null mutants. Our results demonstrated that indeed GK, PEPCK, and PPDK are key players in the gluconeogenesis pathway in Leishmania, although stage-specific differences in their contribution to this pathway were found. GK participates in the entry of glycerol in promastigotes and amastigotes; PEPCK participates in the entry of aspartate in promastigotes, and PPDK is involved in the entry of alanine in amastigotes. Furthermore, the majority of alanine enters into the pathway via decarboxylation of pyruvate in promastigotes, whereas pathway redundancy is suggested for the entry of aspartate in amastigotes. Interestingly, we also found that l-lactate, an abundant glucogenic precursor in mammals, was used by Leishmania amastigotes to synthesize mannogen, entering the pathway through PPDK. On the basis of these new results, we propose a revision in the current model of gluconeogenesis in Leishmania, emphasizing the differences between amastigotes and promastigotes. This work underlines the importance of studying the trypanosomatid intracellular life cycle stages to gain a better understanding of the pathologies caused in humans. PMID:25288791

  4. canal24

    Data.gov (United States)

    California Natural Resource Agency — Canal system center lines in the Central Valley of California and adjacent areas captured from 1:24,000-scale USGS topographic maps. Updates and modifications made...

  5. 40 CFR 209.24 - Default order.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Default order. 209.24 Section 209.24... Orders Issued Under Section 11(d) of the Noise Control Act § 209.24 Default order. (a) Default. Respondent may be found to be in default upon failure to comply with a prehearing or hearing ruling of the...

  6. 24 Pesulan seurantaohjelmisto

    OpenAIRE

    Lumme, Tristan

    2017-01-01

    Insinöörityö tehtiin 24 Pesula Oy:n projektista, joka suoritettiin Helsingissä. Projektin tarkoituksena oli luoda sovellus listaamaan yrityksen toimipisteiden koneet ja niiden käyttökerrat automatisoidusti tietokantaan. 24 Pesula Oy on vuonna 1999 perustettu yritys ja Suomen ainoa itsepalvelupesulaketju. Aikaisemmin koneiden käytönseuranta hoidettiin siten, että käyttödata haettiin paikan päältä lukemalla yksittäisten koneiden lokitiedot. Yrityksen koneissa käytettiin myös Atmelin modifio...

  7. 24 +24 real scalar multiplet in four dimensional N =2 conformal supergravity

    Science.gov (United States)

    Hegde, Subramanya; Lodato, Ivano; Sahoo, Bindusar

    2018-03-01

    Starting from the 48 +48 component multiplet of supercurrents for a rigid N =2 tensor multiplet in four spacetime dimensions, we obtain the transformation of the linearized supergravity multiplet which couples to this supercurrent multiplet. At the linearized level, this 48 +48 component supergravity multiplet decouples into the 24 +24 component linearized standard Weyl multiplet and a 24 +24 component irreducible matter multiplet containing a real scalar field. By a consistent application of the supersymmetry algebra with field-dependent structure constants appropriate to N =2 conformal supergravity, we find the full transformation law for this multiplet in a conformal supergravity background. By performing a suitable field redefinition, we find that the multiplet is a generalization of the flat space multiplet, obtained by Howe et al. in Nucl. Phys. B214, 519 (1983), 10.1016/0550-3213(83)90249-3, to a conformal supergravity background. We also present a set of constraints which can be consistently imposed on this multiplet to obtain a restricted minimal 8 +8 off-shell matter multiplet. We also show, as an example, the precise embedding of the tensor multiplet inside this multiplet.

  8. The analytic renormalization group

    Directory of Open Access Journals (Sweden)

    Frank Ferrari

    2016-08-01

    Full Text Available Finite temperature Euclidean two-point functions in quantum mechanics or quantum field theory are characterized by a discrete set of Fourier coefficients Gk, k∈Z, associated with the Matsubara frequencies νk=2πk/β. We show that analyticity implies that the coefficients Gk must satisfy an infinite number of model-independent linear equations that we write down explicitly. In particular, we construct “Analytic Renormalization Group” linear maps Aμ which, for any choice of cut-off μ, allow to express the low energy Fourier coefficients for |νk|<μ (with the possible exception of the zero mode G0, together with the real-time correlators and spectral functions, in terms of the high energy Fourier coefficients for |νk|≥μ. Operating a simple numerical algorithm, we show that the exact universal linear constraints on Gk can be used to systematically improve any random approximate data set obtained, for example, from Monte-Carlo simulations. Our results are illustrated on several explicit examples.

  9. RADIAL VELOCITY STUDIES OF CLOSE BINARY STARS. XIV

    International Nuclear Information System (INIS)

    Pribulla, Theodor; Rucinski, Slavek M.; DeBond, Heide; De Ridder, Archie; Karmo, Toomas; Thomson, J. R.; Croll, Bryce; Ogloza, Waldemar; Pilecki, Bogumil; Siwak, Michal

    2009-01-01

    Radial velocity (RV) measurements and sine curve fits to the orbital RV variations are presented for 10 close binary systems: TZ Boo, VW Boo, EL Boo, VZ CVn, GK Cep, RW Com, V2610 Oph, V1387 Ori, AU Ser, and FT UMa. Our spectroscopy revealed two quadruple systems, TZ Boo and V2610 Oph, while three stars showing small photometric amplitudes, EL Boo, V1387 Ori, and FT UMa, were found to be triple systems. GK Cep is a close binary with a faint third component. While most of the studied eclipsing systems are contact binaries, VZ CVn and GK Cep are detached or semidetached double-lined binaries, and EL Boo, V1387 Ori, and FT UMa are close binaries of uncertain binary type. The large fraction of triple and quadruple systems found in this sample supports the hypothesis of formation of close binaries in multiple stellar systems; it also demonstrates that low photometric amplitude binaries are a fertile ground for further discoveries of multiple systems.

  10. TOTAL NON-STRUCTURAL CARBOHYDRATE (TNC OF THREE CULTIVARS OF NAPIER GRASS (Pennisetum purpureum AT VEGETATIVE AND REPRODUCTIVE PHASE

    Directory of Open Access Journals (Sweden)

    B. Budiman

    2014-10-01

    Full Text Available An experiment was conducted to determine Total Non-structural carbohydrates (TNC of threecultivars of napier grass (Pennisetum purpureum harvested at vegetative and reproductive phases. Thecultivars tested were Taiwan (Gt, King (Gk and Mott (Gm and arranged in a 3 x 2 of treatments withfour replicates following nested design. The results showed that the highest sugar content (P<0.01 wasfound in Gt cultivar and the lowest was in Gm cultivar. The highest starch content (P<0.01 was found inGk cultivar and the lowest was in Gt cultivar. TNC content of Gt and Gk cultivars were not significantlydifferent, but both were significantly higher (P<0.01 compared with the Gm cultivar. It can beconcluded, that there were differences in TNC between cultivars, however, the TNC content in Gkcultivar was not different with Gt cultivar, while Gm cultivar have the lowest (P<0.01 TNC content. Atreproductive phase all cultivars have higher (P<0.01 TNC and starch content than at vegetative phase

  11. Curiosities at c=24

    International Nuclear Information System (INIS)

    Schellekens, A.N.; Yankielowicz, S.

    1989-01-01

    Two examples of c=24 holomorphic conformal field theories are given that do not correspond to Niemeier lattices or Z 2 twists of Niemeier lattices. One of these examples clarifies a recently discovered modular invariant of F 4 at level 6, which can be interpreted as the 'complement' of SU(3) level 2 within a new c=24 theory. (orig.)

  12. Geneva 24 hours swim

    CERN Document Server

    2003-01-01

    The 18th edition of the Geneva 24 hours swim competition will take place at the Vernets Swimming Pool on the 4th and 5th of October. More information and the results of previous years are given at: http://www.carouge-natation.com/24_heures/home_24_heures.htm Last year, CERN obtained first position in the inter-company category with a total of 152.3 kms swam by 45 participants. We are counting on your support to repeat this excellent performance this year. For those who would like to train, the Livron swimming pool in Meyrin is open as from Monday the 8th September. For further information please do not hesitate to contact us. Gino de Bilio and Catherine Delamare

  13. Geneva 24 Hours Swim

    CERN Document Server

    2003-01-01

    The 18th edition of the Geneva 24 hours swim competition will take place at the Vernets Swimming Pool on the 4th and 5th of October. More information and the results of previous years are given at: http://www.carouge-natation.com/24_heures/home_24_heures.htm Last year, CERN obtained first position in the inter-company category with a total of 152.3 kms swam by 45 participants. We are counting on your support to repeat this excellent performance this year. For those who would like to train, the Livron swimming pool in Meyrin is open as from Monday the 8th September. For further information please do not hesitate to contact us. Gino de Bilio and Catherine Delamare

  14. Purification and Characterization of Taq Polymerase: A 9-Week Biochemistry Laboratory Project for Undergraduate Students

    Science.gov (United States)

    Bellin, Robert M.; Bruno, Mary K.; Farrow, Melissa A.

    2010-01-01

    We have developed a 9-week undergraduate laboratory series focused on the purification and characterization of "Thermus aquaticus" DNA polymerase (Taq). Our aim was to provide undergraduate biochemistry students with a full-semester continuing project simulating a research-like experience, while having each week's procedure focus on a single…

  15. 19 CFR 210.24 - Interlocutory appeals.

    Science.gov (United States)

    2010-04-01

    ... 19 Customs Duties 3 2010-04-01 2010-04-01 false Interlocutory appeals. 210.24 Section 210.24 Customs Duties UNITED STATES INTERNATIONAL TRADE COMMISSION INVESTIGATIONS OF UNFAIR PRACTICES IN IMPORT TRADE ADJUDICATION AND ENFORCEMENT Motions § 210.24 Interlocutory appeals. Rulings by the administrative...

  16. ASA24-Kids (no longer available)

    Science.gov (United States)

    ASA24-Kids-2014 was released in February 2014 and until March 2017, researchers can register new studies in this version of the ASA24® system. Funding is not currently available for a mobile accessible version for kids, such as ASA24-2016.

  17. 25 CFR 225.24 - Environmental studies.

    Science.gov (United States)

    2010-04-01

    ... 25 Indians 1 2010-04-01 2010-04-01 false Environmental studies. 225.24 Section 225.24 Indians... SOLID MINERALS AGREEMENTS Minerals Agreements § 225.24 Environmental studies. (a) The Secretary shall ensure that all environmental studies are prepared as required by the National Environmental Policy Act...

  18. 43 CFR 24.5 - International agreements.

    Science.gov (United States)

    2010-10-01

    ... WILDLIFE POLICY: STATE-FEDERAL RELATIONSHIPS § 24.5 International agreements. (a) International conventions... 43 Public Lands: Interior 1 2010-10-01 2010-10-01 false International agreements. 24.5 Section 24... shall be to recommend that the United States negotiate and accede to only those international agreements...

  19. Gamma Knife Perfexion® radiosurgery and endo diode laser thermotherapy for choroidal melanoma with technical analysis: A case report.

    Science.gov (United States)

    Tsai, Yi-Chieh; Kuo, Chun-Yuan; Lin, Jia-Wei; Yang, Shun-Tai; Lai, Shih-Chung; Tsai, Jo-Ting

    2018-01-01

    Radiosurgery serves an important function in the treatment of patients with intraocular tumors and preserves visual function via organ conservation. Therefore, it is important to ensure the safety and precision of GK-SRS as a primary treatment for intraocular tumors. The present case study described a 57-year-old female with uveal melanoma treated with GK-SRS. Retrobulbar anesthesia following fixation of the treated eye, via the suture of two of the extraocular muscles to the stereotactic frame, was performed to immobilize the eye during treatment. Computed tomography (CT) scans were performed following eye fixation, immediately prior to and following GK-SRS, to validate the accuracy of the tumor localization. The eye movement analysis revealed that the gravity center point deviations of the tumor and lens during treatment were <0.110 mm. At least 95% of the tumor volume was covered by the prescription dose according to three sets of CT images. The patient underwent a trans pars plana vitrectomy owing to a right eye vitreous hemorrhage. A 37-month follow-up assessment revealed tumor shrinkage, and the disappearance of the serous retinal detachments was noted on the basis of ophthalmoscopy and orbital magnetic resonance imaging. No major complications developed during the follow-up period. Using our treatment protocol, GK-SRS is a non-invasive procedure which is used as a brief single fraction treatment for intraocular tumor. The eye fixation method used in the present study has high accuracy.

  20. Thermonuclear reaction rate of 17O(p,γ)18F

    International Nuclear Information System (INIS)

    Fox, C.; Iliadis, C.; Champagne, A.E.; Fitzgerald, R.P.; Longland, R.; Newton, J.; Pollanen, J.; Runkle, R.

    2005-01-01

    The 17 O(p,γ) 18 F and 17 O(p,α) 14 N reactions have a profound influence on hydrogen-burning nucleosynthesis in a number of stellar sites, including red giants, asymptotic giant branch (AGB) stars, massive stars, and classical novae. Previously evaluated thermonuclear rates for both reactions carry large uncertainties. We investigated the proton-capture reaction on 17 O in the bombarding energy range of E p lab = 180-540 keV. We observed a previously undiscovered resonance at E R lab = 193.2 ± 0.9 keV. The resonance strength amounts to (ωγ) pγ (1.2±0.2)x10 -6 eV. With this value, the uncertainties of the 17 O(p,γ) 18 F reaction rates are reduced by orders of magnitude in the peak temperature range of classical novae (T=0.1-0.4 GK). We also report on a reevaluation of the 17 O(p,γ) 18 F reaction rates at lower temperatures that are pertinent to red giants, AGB stars, or massive stars. The present work establishes the 17 O(p,γ) 18 F reaction rates over a temperature range of T= 0.01-1.5 GK with statistical uncertainties of 10-50%. The new recommended reaction rates deviate from the previously accepted values by an order of magnitude around T≅0.2 GK and by factors of 2-3 at T < 0.1 GK

  1. Gluconeogenesis in Leishmania mexicana: contribution of glycerol kinase, phosphoenolpyruvate carboxykinase, and pyruvate phosphate dikinase.

    Science.gov (United States)

    Rodriguez-Contreras, Dayana; Hamilton, Nicklas

    2014-11-21

    Gluconeogenesis is an active pathway in Leishmania amastigotes and is essential for their survival within the mammalian cells. However, our knowledge about this pathway in trypanosomatids is very limited. We investigated the role of glycerol kinase (GK), phosphoenolpyruvate carboxykinase (PEPCK), and pyruvate phosphate dikinase (PPDK) in gluconeogenesis by generating the respective Leishmania mexicana Δgk, Δpepck, and Δppdk null mutants. Our results demonstrated that indeed GK, PEPCK, and PPDK are key players in the gluconeogenesis pathway in Leishmania, although stage-specific differences in their contribution to this pathway were found. GK participates in the entry of glycerol in promastigotes and amastigotes; PEPCK participates in the entry of aspartate in promastigotes, and PPDK is involved in the entry of alanine in amastigotes. Furthermore, the majority of alanine enters into the pathway via decarboxylation of pyruvate in promastigotes, whereas pathway redundancy is suggested for the entry of aspartate in amastigotes. Interestingly, we also found that l-lactate, an abundant glucogenic precursor in mammals, was used by Leishmania amastigotes to synthesize mannogen, entering the pathway through PPDK. On the basis of these new results, we propose a revision in the current model of gluconeogenesis in Leishmania, emphasizing the differences between amastigotes and promastigotes. This work underlines the importance of studying the trypanosomatid intracellular life cycle stages to gain a better understanding of the pathologies caused in humans. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

  2. Radiosurgery of pituitary adenomas

    International Nuclear Information System (INIS)

    Kida, Yoshihisa

    2008-01-01

    The efficacy and role of gamma knife (GK) in the treatment of various pituitary adenomas are described on author's experience and discussed with literature. GK subjects are 328 patients (M 126/F 202, av. age of 47.8 y) in author's hospital, and satisfactory follow-up (32-44 mo) for evaluation has been possible in 253 cases, who had tumors non-functional (129 cases), producing ACTH (23), HGH (70) and PRL (31). Stereotactic GK radiosurgery is done with navigation by Gamma Plan based on enhanced MRI images at various doses, and evaluation in the follow-up period is performed by hormonal levels and MRI which give efficacy of complete response (CR), partial response (PR), MR and standard deviation (SD)/ progressive disease (PD) on the tumor size. The overall tumor control rate is found to be 95-100%. Effectiveness (CR and PR) is found as high as 77.4% in PRL-producing tumor (marginal dose 14-32 Gy), 65% in non-functioning (15-25 Gy), 61% in ACTH (19-30 Gy) and 60% in GH (19-31 Gy), of which tendency is similar to that in literature. Even in ACTH-producing tumor, low ACTH and cortisol levels persisted with tendency of improved obese and hypertensive symptoms. GK radiosurgery has limitations in the tissue size and distance between the tumor and optic nerve/chiasm, but for the enough small tumor, it gives satisfactorily long term efficacy. (R.T.)

  3. Interdependency between mechanical parameters and afferent nerve discharge in remodeled diabetic Goto-Kakizaki rat intestine.

    Science.gov (United States)

    Zhao, Jingbo; Yang, Jian; Liao, Donghua; Gregersen, Hans

    2017-01-01

    Gastrointestinal disorders are very common in diabetic patients, but the pathogenesis is still not well understood. Peripheral afferent nerves may be involved due to the complex regulation of gastrointestinal function by the enteric nervous system. We aimed to characterize the stimulus-response function of afferent fibers innervating the jejunum in the Goto-Kakizaki (GK) type 2 diabetic rat model. A key question is whether changes in afferent firing arise from remodeled tissue or from adaptive afferent processes. Seven 32-week-old male GK rats and seven age-matched normal Wistar rats were studied. Firing from mesenteric afferent nerves was recorded in excised jejunal segments of seven GK rats and seven normal Wistar rats during ramp test, stress relaxation test, and creep test. The circumferential stress-strain, spike rate increase ratio (SRIR), and single unit firing rates were calculated for evaluation of interdependency of the mechanical stimulations and the afferent nerve discharge. Elevated sensitivity to mechanical stimuli was found for diabetic nerve bundles and single unit activity ( P <0.05). The stress relaxed less in the diabetic intestinal segment ( P <0.05). Linear association between SRIR and the thickness of circumferential muscle layer was found at high stress levels as well as for SRIR and the glucose level. Altered viscoelastic properties and elevated mechanosensitivity were found in the GK rat intestine. The altered nerve signaling is related to muscle layer remodeling and glucose levels and may contribute to gastrointestinal symptoms experienced by diabetic patients.

  4. 12 CFR 24.4 - Investment limits.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 1 2010-01-01 2010-01-01 false Investment limits. 24.4 Section 24.4 Banks and... ENTITIES, COMMUNITY DEVELOPMENT PROJECTS, AND OTHER PUBLIC WELFARE INVESTMENTS § 24.4 Investment limits. (a) Limits on aggregate outstanding investments. A national bank's aggregate outstanding investments under...

  5. 45 CFR 99.24 - Evidentiary purpose.

    Science.gov (United States)

    2010-10-01

    ... 45 Public Welfare 1 2010-10-01 2010-10-01 false Evidentiary purpose. 99.24 Section 99.24 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION PROCEDURE FOR HEARINGS FOR THE CHILD CARE AND DEVELOPMENT FUND Hearing Procedures § 99.24 Evidentiary purpose. The purpose of the...

  6. 7 CFR 3019.24 - Program income.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 15 2010-01-01 2010-01-01 false Program income. 3019.24 Section 3019.24 Agriculture... Management § 3019.24 Program income. (a) Federal awarding agencies shall apply the standards set forth in this section in requiring recipient organizations to account for program income related to projects...

  7. 12 CFR 561.24 - Immediate family.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Immediate family. 561.24 Section 561.24 Banks and Banking OFFICE OF THRIFT SUPERVISION, DEPARTMENT OF THE TREASURY DEFINITIONS FOR REGULATIONS AFFECTING ALL SAVINGS ASSOCIATIONS § 561.24 Immediate family. The term immediate family of any natural...

  8. 49 CFR 176.24 - Shipping papers.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 2 2010-10-01 2010-10-01 false Shipping papers. 176.24 Section 176.24... Requirements § 176.24 Shipping papers. (a) A person may not accept a hazardous material for transportation or transport a hazardous material by vessel unless that person has received a shipping paper prepared in...

  9. 49 CFR 174.24 - Shipping papers.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 2 2010-10-01 2010-10-01 false Shipping papers. 174.24 Section 174.24... Requirements § 174.24 Shipping papers. (a) A person may not accept a hazardous material for transportation or transport a hazardous material by rail unless that person receives a shipping paper prepared in accordance...

  10. 33 CFR 2.24 - Internal waters.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Internal waters. 2.24 Section 2.24 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY GENERAL JURISDICTION Jurisdictional Terms § 2.24 Internal waters. (a) With respect to the United States, internal waters means the...

  11. 34 CFR 74.24 - Program income.

    Science.gov (United States)

    2010-07-01

    ... 34 Education 1 2010-07-01 2010-07-01 false Program income. 74.24 Section 74.24 Education Office of... Program Management § 74.24 Program income. (a) The Secretary applies the standards contained in this section in requiring recipient organizations to account for program income related to projects financed in...

  12. 22 CFR 145.24 - Program income.

    Science.gov (United States)

    2010-04-01

    ... 22 Foreign Relations 1 2010-04-01 2010-04-01 false Program income. 145.24 Section 145.24 Foreign....24 Program income. (a) The Department shall apply the standards set forth in this section in requiring recipient organizations to account for program income related to projects financed in whole or in...

  13. 49 CFR 19.24 - Program income.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 1 2010-10-01 2010-10-01 false Program income. 19.24 Section 19.24 Transportation... Requirements § 19.24 Program income. (a) Federal awarding agencies shall apply the standards set forth in this section in requiring recipient organizations to account for program income related to projects financed in...

  14. 22 CFR 518.24 - Program income.

    Science.gov (United States)

    2010-04-01

    ... 22 Foreign Relations 2 2010-04-01 2010-04-01 true Program income. 518.24 Section 518.24 Foreign... Financial and Program Management § 518.24 Program income. (a) Federal awarding agencies shall apply the standards set forth in this section in requiring recipient organizations to account for program income...

  15. 20 CFR 435.24 - Program income.

    Science.gov (United States)

    2010-04-01

    ... 20 Employees' Benefits 2 2010-04-01 2010-04-01 false Program income. 435.24 Section 435.24... ORGANIZATIONS Post-Award Requirements Financial and Program Management § 435.24 Program income. (a) Introduction... for program income related to projects financed in whole or in part with Federal funds. (b) Use of...

  16. 28 CFR 70.24 - Program income.

    Science.gov (United States)

    2010-07-01

    ... 28 Judicial Administration 2 2010-07-01 2010-07-01 false Program income. 70.24 Section 70.24...-PROFIT ORGANIZATIONS Post-Award Requirements Financial and Program Management § 70.24 Program income. (a... income related to projects financed in whole or in part with Department funds. (b) Except as provided in...

  17. 23 CFR 1200.24 - Program income.

    Science.gov (United States)

    2010-04-01

    ... 23 Highways 1 2010-04-01 2010-04-01 false Program income. 1200.24 Section 1200.24 Highways... Implementation and Management of the Highway Safety Program § 1200.24 Program income. (a) Inclusions. Program income includes income from fees for services performed, from the use or rental of real or personal...

  18. 27 CFR 24.217 - Vinegar stock.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Vinegar stock. 24.217 Section 24.217 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS WINE Production of Other Than Standard Wine § 24.217 Vinegar stock. Vinegar...

  19. 15 CFR 24.41 - Financial reporting.

    Science.gov (United States)

    2010-01-01

    ... 15 Commerce and Foreign Trade 1 2010-01-01 2010-01-01 false Financial reporting. 24.41 Section 24..., Records, Retention, and Enforcement § 24.41 Financial reporting. (a) General. (1) Except as provided in... by OMB, for: (i) Submitting financial reports to Federal agencies, or (ii) Requesting advances or...

  20. 24 CFR 1000.24 - If an Indian tribe assumes environmental review responsibility, how will HUD assist the Indian...

    Science.gov (United States)

    2010-04-01

    ...? 1000.24 Section 1000.24 Housing and Urban Development Regulations Relating to Housing and Urban... URBAN DEVELOPMENT NATIVE AMERICAN HOUSING ACTIVITIES General § 1000.24 If an Indian tribe assumes...

  1. The Kepler Dichotomy in Planetary Disks: Linking Kepler Observables to Simulations of Late-stage Planet Formation

    Science.gov (United States)

    Moriarty, John; Ballard, Sarah

    2016-11-01

    NASA’s Kepler Mission uncovered a wealth of planetary systems, many with planets on short-period orbits. These short-period systems reside around 50% of Sun-like stars and are similarly prevalent around M dwarfs. Their formation and subsequent evolution is the subject of active debate. In this paper, we simulate late-stage, in situ planet formation across a grid of planetesimal disks with varying surface density profiles and total mass. We compare simulation results with observable characteristics of the Kepler sample. We identify mixture models with different primordial planetesimal disk properties that self-consistently recover the multiplicity, radius, period and period ratio, and duration ratio distributions of the Kepler planets. We draw three main conclusions. (1) We favor a “frozen-in” narrative for systems of short-period planets, in which they are stable over long timescales, as opposed to metastable. (2) The “Kepler dichotomy,” an observed phenomenon of the Kepler sample wherein the architectures of planetary systems appear to either vary significantly or have multiple modes, can naturally be explained by formation within planetesimal disks with varying surface density profiles. Finally, (3) we quantify the nature of the “Kepler dichotomy” for both GK stars and M dwarfs, and find that it varies with stellar type. While the mode of planet formation that accounts for high multiplicity systems occurs in 24% ± 7% of planetary systems orbiting GK stars, it occurs in 63% ± 16% of planetary systems orbiting M dwarfs.

  2. Introduction to Communication Systems

    Science.gov (United States)

    2014-01-17

    search breakthroughs in coding and modulation can be quickly transitioned into products. This leads to economies of scale that have been critical to the...implementation of its transceiver functionalities in DSP, which leverages the economies of scale of digital computation (Moore’s “law”). For T large...1)!e − gIg ≥0 (8.92) FG(g) = P [G ≤ g] = e−g ∞ ∑ k=N gk k! = 1− e−g N−1 ∑ k=0 gk k! , g ≥ 0 (8.93) (a) Use the preceding results to compute the

  3. An intermolecular binding mechanism involving multiple LysM domains mediates carbohydrate recognition by an endopeptidase

    DEFF Research Database (Denmark)

    Wong, Mei Mei Jaslyn Elizabeth; Midtgaard, Søren Roi; Gysel, Kira

    2015-01-01

    of multiple LysM domains in substrate binding has so far lacked support from high-resolution structures of ligand-bound complexes. Here, a structural study of the Thermus thermophilus NlpC/P60 endopeptidase containing two LysM domains is presented. The crystal structure and small-angle X-ray scattering...

  4. 45 CFR 74.24 - Program income.

    Science.gov (United States)

    2010-10-01

    ... 45 Public Welfare 1 2010-10-01 2010-10-01 false Program income. 74.24 Section 74.24 Public Welfare... COMMERCIAL ORGANIZATIONS Post-Award Requirements Financial and Program Management § 74.24 Program income. (a) The standards set forth in this section shall be used to account for program income related to...

  5. 45 CFR 2543.24 - Program income.

    Science.gov (United States)

    2010-10-01

    ... 45 Public Welfare 4 2010-10-01 2010-10-01 false Program income. 2543.24 Section 2543.24 Public... ORGANIZATIONS Post-Award Requirements Financial and Program Management § 2543.24 Program income. (a) Federal... to account for program income related to projects financed in whole or in part with Federal funds. (b...

  6. 30 CFR 282.24 - Mining Plan.

    Science.gov (United States)

    2010-07-01

    ... 30 Mineral Resources 2 2010-07-01 2010-07-01 false Mining Plan. 282.24 Section 282.24 Mineral... § 282.24 Mining Plan. All OCS mineral development and production activities shall be conducted in accordance with a Mining Plan submitted by the lessee and approved by the Director. A Mining Plan shall...

  7. 47 CFR 24.237 - Interference protection.

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 2 2010-10-01 2010-10-01 false Interference protection. 24.237 Section 24.237... SERVICES Broadband PCS § 24.237 Interference protection. (a) All licensees are required to coordinate their... protection criterion shall be such that the interfering signal will not produce more than 1.0 dB degradation...

  8. 6 CFR 13.24 - Protective order.

    Science.gov (United States)

    2010-01-01

    ... 6 Domestic Security 1 2010-01-01 2010-01-01 false Protective order. 13.24 Section 13.24 Domestic Security DEPARTMENT OF HOMELAND SECURITY, OFFICE OF THE SECRETARY PROGRAM FRAUD CIVIL REMEDIES § 13.24 Protective order. (a) A party or a prospective witness or deponent may file a motion for a protective order...

  9. 30 CFR 36.24 - Engine joints.

    Science.gov (United States)

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Engine joints. 36.24 Section 36.24 Mineral... Construction and Design Requirements § 36.24 Engine joints. (a) Cylinder head. The joint between the cylinder head and block of the engine shall be fitted with a metal or metal-clad gasket satisfactory to MSHA...

  10. 15 CFR 14.24 - Program income.

    Science.gov (United States)

    2010-01-01

    ... 15 Commerce and Foreign Trade 1 2010-01-01 2010-01-01 false Program income. 14.24 Section 14.24... ORGANIZATIONS Post-Award Requirements Financial and Program Management § 14.24 Program income. (a) The standards... income related to projects financed in whole or in part with Federal funds. (b) Except as provided in...

  11. 29 CFR 95.24 - Program income.

    Science.gov (United States)

    2010-07-01

    ... 29 Labor 1 2010-07-01 2010-07-01 true Program income. 95.24 Section 95.24 Labor Office of the... Program Management § 95.24 Program income. (a) Except as provided in paragraph (e) of this section, program income earned during the project period shall be retained by the recipient and added to funds...

  12. 15 CFR 24.25 - Program income.

    Science.gov (United States)

    2010-01-01

    ... 15 Commerce and Foreign Trade 1 2010-01-01 2010-01-01 false Program income. 24.25 Section 24.25... Administration § 24.25 Program income. (a) General. Grantees are encouraged to earn income to defray program costs. Program income includes income from fees for services performed, from the use or rental of real...

  13. 27 CFR 24.266 - Inventory losses.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Inventory losses. 24.266... OF THE TREASURY LIQUORS WINE Losses of Wine § 24.266 Inventory losses. (a) General. The proprietor... reported as required by § 24.313. (b) Bulk wine losses. The physical inventory of bulk wine will determine...

  14. Atrial Fibrillation Detection During 24-Hour Ambulatory Blood Pressure Monitoring: Comparison With 24-Hour Electrocardiography.

    Science.gov (United States)

    Kollias, Anastasios; Destounis, Antonios; Kalogeropoulos, Petros; Kyriakoulis, Konstantinos G; Ntineri, Angeliki; Stergiou, George S

    2018-07-01

    This study assessed the diagnostic accuracy of a novel 24-hour ambulatory blood pressure (ABP) monitor (Microlife WatchBP O3 Afib) with implemented algorithm for automated atrial fibrillation (AF) detection during each ABP measurement. One hundred subjects (mean age 70.6±8.2 [SD] years; men 53%; hypertensives 85%; 17 with permanent AF; 4 paroxysmal AF; and 79 non-AF) had simultaneous 24-hour ABP monitoring and 24-hour Holter monitoring. Among a total of 6410 valid ABP readings, 1091 (17%) were taken in ECG AF rhythm. In reading-to-reading ABP analysis, the sensitivity, specificity, and accuracy of ABP monitoring in detecting AF were 93%, 87%, and 88%, respectively. In non-AF subjects, 12.8% of the 24-hour ABP readings indicated false-positive AF, of whom 27% were taken during supraventricular premature beats. There was a strong association between the proportion of false-positive AF readings and that of supraventricular premature beats ( r =0.67; P ABP monitoring had 100%/85% sensitivity/specificity (area under the curve 0.91; P ABP monitor with AF detector has high sensitivity and moderate specificity for AF screening during routine ABP monitoring. Thus, in elderly hypertensives, a 24-hour ABP recording with at least 26% of the readings suggesting AF indicates a high probability for AF diagnosis and should be regarded as an indication for performing 24-hour Holter monitoring. © 2018 American Heart Association, Inc.

  15. A novel HLA-A*24 allele, A*24:231, was identified by sequencing-based typing.

    Science.gov (United States)

    Li, G Y; Liu, Y Y; Wu, K L; Tang, Z H

    2018-05-22

    HLA-A*24:231 has 1 nucleotide change from HLA-A*24:02:01:01 at position 784 G>C This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  16. Activities of asymmetric dimethylarginine-related enzymes in white adipose tissue are associated with circulating lipid biomarkers

    Directory of Open Access Journals (Sweden)

    Iwasaki Hiroaki

    2012-04-01

    Full Text Available Abstract Background Asymmetric NG,NG-dimethylarginine (ADMA, an endogenous inhibitor of nitric oxide synthase, is regulated by the enzymatic participants of synthetic and metabolic processes, i.e., type I protein N-arginine methyltransferase (PRMT and dimethylarginine dimethylaminohydrolase (DDAH. Previous reports have demonstrated that circulating ADMA levels can vary in patients with type 1 and type 2 diabetes mellitus (T2DM. White adipose tissue expresses the full enzymatic machinery necessary for ADMA production and metabolism; however, modulation of the activities of adipose ADMA-related enzymes in T2DM remains to be determined. Methods A rodent model of T2DM using 11- and 20-week old Goto-Kakizaki (GK rats was used. The expression and catalytic activity of PRMT1 and DDAH1 and 2 in the white adipose tissues (periepididymal, visceral and subcutaneous fats and femur skeletal muscle tissue were determined by immunoblotting, in vitro methyltransferase and in vitro citrulline assays. Results Non-obese diabetic GK rats showed low expression and activity of adipose PRMT1 compared to age-matched Wistar controls. Adipose tissues from the periepididymal, visceral and subcutaneous fats of GK rats had high DDAH1 expression and total DDAH activity, whereas the DDAH2 expression was lowered below the control value. This dynamic of ADMA-related enzymes in white adipose tissues was distinct from that of skeletal muscle tissue. GK rats had lower levels of serum non-esterified fatty acids (NEFA and triglycerides (TG than the control rats. In all subjects the adipose PRMT1 and DDAH activities were statistically correlated with the levels of serum NEFA and TG. Conclusion Activities of PRMT1 and DDAH in white adipose tissues were altered in diabetic GK rats in an organ-specific manner, which was reflected in the serum levels of NEFA and TG. Changes in adipose ADMA-related enzymes might play a part in the function of white adipose tissue.

  17. 24 CFR 511.74 - Audit.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 3 2010-04-01 2010-04-01 false Audit. 511.74 Section 511.74... CLEARANCE AND URBAN RENEWAL RENTAL REHABILITATON GRANT PROGRAM Grant Administration § 511.74 Audit. The... audits in accordance with 24 CFR part 44. ...

  18. 12 CFR 228.24 - Service test.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 3 2010-01-01 2010-01-01 false Service test. 228.24 Section 228.24 Banks and... REINVESTMENT (REGULATION BB) Standards for Assessing Performance § 228.24 Service test. (a) Scope of test. The service test evaluates a bank's record of helping to meet the credit needs of its assessment area(s) by...

  19. 15 CFR 24.32 - Equipment.

    Science.gov (United States)

    2010-01-01

    ... awarding agency. (d) Management requirements. Procedures for managing equipment (including replacement... return. (e) Disposition. When original or replacement equipment acquired under a grant or subgrant is no... 15 Commerce and Foreign Trade 1 2010-01-01 2010-01-01 false Equipment. 24.32 Section 24.32...

  20. 7 CFR 948.24 - Safeguards.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false Safeguards. 948.24 Section 948.24 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing Agreements... entering trade channels other than those authorized by regulations and by such rules as may be necessary...

  1. The nucleotide sequence of 5S ribosomal RNA from Micrococcus lysodeikticus.

    Science.gov (United States)

    Hori, H; Osawa, S; Murao, K; Ishikura, H

    1980-01-01

    The nucleotide sequence of ribosomal 5S RNA from Micrococcus lysodeikticus is pGUUACGGCGGCUAUAGCGUGGGGGAAACGCCCGGCCGUAUAUCGAACCCGGAAGCUAAGCCCCAUAGCGCCGAUGGUUACUGUAACCGGGAGGUUGUGGGAGAGUAGGUCGCCGCCGUGAOH. When compared to other 5S RNAs, the sequence homology is greatest with Thermus aquaticus, and these two 5S RNAs reveal several features intermediate between those of typical gram-positive bacteria and gram-negative bacteria. PMID:6780979

  2. Microcircuit Device Reliability. Digital Evaluation and Failure Analysis Data. Parts 1 and 2, Summer 1980

    Science.gov (United States)

    1980-01-01

    C - 7403 24 :0170C v V ISCTC :o IO a - TVYPCYC :-06r 50 ISO 1±8-U3 42: : 0: ,I~ 4 : I VCVC -2010C MOI- 0ST C"’IO C6SC -MS-883 - 42: 0: - - :V 96i’&u...FINE LK :HE 5.S-8 : MS- 8583 : 64: 60l 1 : : :V . :60 -h-1N : 101-4 A : . : :GkOSSLK :FLUOR 125C Ih-S-3 64: 0, * . : V : 31 : lu14 C : T.I. : E...5.4 . : : : 1 035110J SVARIOUS : DIP :5 .35/76 :.VIS INS :±150: : 0* ± 74122 ± 14 :0/70C : I : ± N.E. "" . . - :S4F E4 :025C 070C ISO :5± 0

  3. 30 CFR 44.24 - Discovery.

    Science.gov (United States)

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Discovery. 44.24 Section 44.24 Mineral... Discovery. Parties shall be governed in their conduct of discovery by appropriate provisions of the Federal... discovery. Alternative periods of time for discovery may be prescribed by the presiding administrative law...

  4. 15 CFR 24.4 - Applicability.

    Science.gov (United States)

    2010-01-01

    ... 15 Commerce and Foreign Trade 1 2010-01-01 2010-01-01 false Applicability. 24.4 Section 24.4 Commerce and Foreign Trade Office of the Secretary of Commerce UNIFORM ADMINISTRATIVE REQUIREMENTS FOR...; Maternal and Child Health Services; Social Services; Low-Income Home Energy Assistance; States' Program of...

  5. Clinical Value of CD24 Expression in Retinoblastoma

    Directory of Open Access Journals (Sweden)

    Jia Li

    2012-01-01

    Full Text Available Background. The expression of CD24 has been detected in a wide variety of human malignancies. Downregulation of CD24 inhibits proliferation and induces apoptosis in tumor cells, whereas its upregulation increases tumor growth and metastasis. However, no data on CD24 protein levels in retinoblastoma are available, and the mechanism of CD24 involvement in retinoblastoma progress has not been elucidated. The aim of this study was to explore the expression profile of CD24 in the retinoblastoma tumor samples and to correlate with clinicopathological parameters. Methods. Immunohistochemistry was performed for CD24 on the archival paraffin sections of retinoblastoma and correlated with clinicopathological features. Western blotting was performed to confirm immunoreactivity results. Results. CD24 immunoreactivity was observed in 72.0% (36/50 of the retinoblastoma specimens. Among the 35 low-risk tumors, CD24 was expressed in 62.9% (22/35 tumors and among the 15 high-risk tumors, CD24 was expressed in 93.3% (14/15 tumors. High-risk tumors showed significantly increased expression of CD24 compared to tumors with low-risk (<0.05. Conclusions. This is the first correlation between CD24 expression and histopathology in human retinoblastoma. Our study showed increased expression of CD24 in high risk tumors compared to low risk tumors. Further functional studies are required to explore the role of CD24 in retinoblastoma.

  6. Long-term follow-up results of combination therapy of surgery and gamma knife on pituitary tumor

    International Nuclear Information System (INIS)

    Ikeda, Hidetoshi; Yoshimoto, Takashi; Shirokura, Hideshi

    1997-01-01

    Usefulness of the combination therapy for invasive pituitary tumor with surgery and gamma knife (GK) was evaluated on 17 cases followed for >2 years. Tumors involved ACTH cell adenoma, GH cell adenoma, mixed GH and PRL, purihormonal adenoma, gonadotrophic cell adenoma, GH cell adenoma and null cell adenoma, which were divided into I and II group since adverse effects by GK on the normal pituitary tissues could be evaluated according to tumor growth and abnormal hormone secretion. Irradiation was carried out to make the marginal dose of around 30 (15-35) Gy and center dose of 25-70 Gy on the gland, and marginal dose of <10 Gy on optic nerve. After GK, follow-up was done by pituitary hormone values, tests for sight and visual field and MRI examination. Hypopituitarism was seen in 67% with 100% remission of Cushing's disease. MRI revealed that the adenoma changed to fibrosis (type 1) with increasing Gd-enhancement or to cystic necrosis (type 2) without Gd-enhancement. Values for GH, cortisol and ACTH turned to normal ones. Thus the combination therapy was found useful. (K.H.)

  7. A treatment strategy for arteriovenous malformations of the brain based on results of gamma knife surgery experienced between 1991 and 1995

    International Nuclear Information System (INIS)

    Inoue, Hiroshi K.; Nagaseki, Yoshishige; Nishi, Hideo; Sanada, Tomoyuki; Tachikawa, Yuichi; Nishimatsu, Terutaka; Ohye, Chihiro; Andou, Yoshitaka; Naitou, Isao

    2001-01-01

    We investigated a treatment strategy for arteriovenous malformations (AVMs) of the brain based on results of 107 patients treated with Gamma Knife (GK) surgery between 1991 and 1995. Total obliteration was obtained in 80.5%. The obliteration rate was 82.1% in Grade 3 of Spetzler-Martin scale, 75.0% in Grade 4, 50.0% in Grade 5, and 66.7% in Grade 6. The obliteration rate of slow and low flow AVMs (moya type) within 12 months (65.5%) was significantly higher than that of rapid and/or high flow AVMs (shunt, mixed type). Six patients had bleeding 7 to 42 months after GK surgery. The annual rate of bleeding in accumulated years was 2.3%. Adverse effects appeared in 2 (1.9%) of 107 patients, and both patients had AVMs in the internal capsule. We concluded that microsurgery is recommended for AVMs operable without morbidity, and GK surgery is recommended for AVMs in functional areas, and that intravascular embolization is required for large, rapid and/or high flow AVMs, especially those with hemorrhagic risk factors. (author)

  8. High dietary fat-induced obesity in Wistar rats and type 2 diabetes in nonobese Goto-Kakizaki rats differentially affect retinol binding protein 4 expression and vitamin A metabolism.

    Science.gov (United States)

    Shirai, Tomomi; Shichi, Yuta; Sato, Miyuki; Tanioka, Yuri; Furusho, Tadasu; Ota, Toru; Tadokoro, Tadahiro; Suzuki, Tsukasa; Kobayashi, Ken-Ichi; Yamamoto, Yuji

    2016-03-01

    Obesity is a major risk factor for type 2 diabetes, which is caused mainly by insulin resistance. Retinol binding protein 4 (RBP4) is the only specific transport protein for retinol in the serum. RBP4 level is increased in the diabetic state and high-fat condition, indicating that retinol metabolism may be affected under these conditions. However, the precise effect of diabetes and high fat-induced obesity on retinol metabolism is unknown. In this study, we examined differences in retinol metabolite levels in rat models of diet-induced obesity and type 2 diabetes (Goto-Kakizaki [GK] rat). Four-week-old male Wistar and GK rats were given either a control diet (AIN-93G) or a high-fat diet (HFD, 40% fat kJ). After 15 weeks of feeding, the RBP4 levels increased by 2-fold in the serum of GK rats but not HFD-fed rats. The hepatic retinol concentration of HFD-fed rats was approximately 50% that of the controls (P type 2 diabetes mellitus. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. 24 CFR 597.3 - Definitions.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 3 2010-04-01 2010-04-01 false Definitions. 597.3 Section 597.3... COMMUNITY FACILITIES URBAN EMPOWERMENT ZONES AND ENTERPRISE COMMUNITIES: ROUND ONE DESIGNATIONS General Provisions § 597.3 Definitions. The terms HUD and Secretary are defined in 24 CFR part 5. Designation means...

  10. 24 CFR 941.209 - Audit.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Audit. 941.209 Section 941.209... HOUSING DEVELOPMENT PHA Eligibility and Program Requirements § 941.209 Audit. All PHAs that receive funds under this part for the development of low-income housing shall comply with audit requirements in 24 CFR...

  11. 24 CFR 574.650 - Audit.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 3 2010-04-01 2010-04-01 false Audit. 574.650 Section 574.650... COMMUNITY FACILITIES HOUSING OPPORTUNITIES FOR PERSONS WITH AIDS Other Federal Requirements § 574.650 Audit... must provide for audits in accordance with 24 CFR part 44. A nonprofit organization that is a grantee...

  12. 24 CFR 884.124 - Audit.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Audit. 884.124 Section 884.124... HOUSING PROJECTS Applicability, Scope and Basic Policies § 884.124 Audit. (a) Where a State or local..., receiving financial assistance under this part, the audit requirements in 24 CFR part 44 shall apply. (b...

  13. 24 CFR 882.124 - Audit.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Audit. 882.124 Section 882.124...) SECTION 8 MODERATE REHABILITATION PROGRAMS Applicability, Scope and Basic Policies § 882.124 Audit. PHAs receiving financial assistance under this part are subject to audit requirements in 24 CFR part 44. [50 FR...

  14. 24 CFR 214.500 - Audit.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 2 2010-04-01 2010-04-01 false Audit. 214.500 Section 214.500... PROGRAM Other Federal Requirements § 214.500 Audit. Housing counseling grant recipients and subrecipients shall be subject to the audit requirements contained in 24 CFR parts 84 and 85. HUD must be provided a...

  15. 27 CFR 24.80 - General.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false General. 24.80 Section 24.80 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE..., agricultural wine, and other than standard wine (except distilling material or vinegar stock) are to be made...

  16. 27 CFR 24.146 - Bonds.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Bonds. 24.146 Section 24.146 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE.... (c) Wine vinegar plant bond. The proprietor of a wine vinegar plant who withdraws wine from a bonded...

  17. Gamma Knife radiosurgery for the treatment of intracranial dural arteriovenous fistulas

    Science.gov (United States)

    Dmytriw, Adam A; Schwartz, Michael L; Cusimano, Michael D; Mendes Pereira, Vitor; Krings, Timo; Tymianski, Michael; Radovanovic, Ivan

    2016-01-01

    Background Intracranial dural arteriovenous fistulae (DAVF) may present a treatment challenge. Endovascular embolization is in most cases the first line of treatment but does not always achieve cure. Gamma Knife (GK) radiosurgery represents an alternative treatment option, and the purpose of this study was to further evaluate its utility. Methods We reviewed all cases of DAVF treated between 2009 and 2016 at our institution with GK radiosurgery independently, or following failed/refused endovascular or surgical management. Patients’ clinical files, radiological images, catheter angiograms, and surgical DAVF disconnection reports were retrospectively reviewed. Results Sixteen DAVF (14 patients) treated by GK radiosurgery were identified. Eleven fistulae were aggressive and five were benign. Marginal doses ranged from 15 to 25 Gy. Target volumes ranged from 0.04 to 4.47 cm3. In all symptomatic patients, GK treatment resulted in symptom palliation. In 13/15 lesions, cure of symptoms (86.0%) was reported. One lesion was asymptomatic. Angiographic cure was achieved in eight cases (50%), small residual DAVF occurred in four, and four were unchanged. One patient developed headache that resolved at one year. No hemorrhage occurred during the follow-up period. There was no significant association between Borden type and cure rate. Prior failed endovascular treatment and small target volume were associated with lower rates of cure. Conclusions Stereotactic radiosurgery is viable treatment for DAVF. It is very effective in palliating symptoms as a de novo approach or adjunctive to endovascular therapy. In our experience it is only somewhat effective in achieving complete angiographic cure. PMID:28156167

  18. Functional Coding Variation in Recombinant Inbred Mouse Lines Reveals Novel Serotonin Transporter-Associated Phenotypes

    Energy Technology Data Exchange (ETDEWEB)

    Carneiro, Ana [Vanderbilt University; Airey, David [University of Tennessee Health Science Center, Memphis; Thompson, Brent [Vanderbilt University; Zhu, C [Vanderbilt University; Rinchik, Eugene M [ORNL; Lu, Lu [University of Tennessee Health Science Center, Memphis; Chesler, Elissa J [ORNL; Erikson, Keith [University of North Carolina; Blakely, Randy [Vanderbilt University

    2009-01-01

    The human serotonin (5-hydroxytryptamine, 5-HT) transporter (hSERT, SLC6A4) figures prominently in the etiology or treatment of many prevalent neurobehavioral disorders including anxiety, alcoholism, depression, autism and obsessive-compulsive disorder (OCD). Here we utilize naturally occurring polymorphisms in recombinant inbred (RI) lines to identify novel phenotypes associated with altered SERT function. The widely used mouse strain C57BL/6J, harbors a SERT haplotype defined by two nonsynonymous coding variants (Gly39 and Lys152 (GK)). At these positions, many other mouse lines, including DBA/2J, encode Glu39 and Arg152 (ER haplotype), assignments found also in hSERT. Synaptosomal 5-HT transport studies revealed reduced uptake associated with the GK variant. Heterologous expression studies confirmed a reduced SERT turnover rate for the GK variant. Experimental and in silico approaches using RI lines (C57Bl/6J X DBA/2J=BXD) identifies multiple anatomical, biochemical and behavioral phenotypes specifically impacted by GK/ER variation. Among our findings are multiple traits associated with anxiety and alcohol consumption, as well as of the control of dopamine (DA) signaling. Further bioinformatic analysis of BXD phenotypes, combined with biochemical evaluation of SERT knockout mice, nominates SERT-dependent 5-HT signaling as a major determinant of midbrain iron homeostasis that, in turn, dictates ironregulated DA phenotypes. Our studies provide a novel example of the power of coordinated in vitro, in vivo and in silico approaches using murine RI lines to elucidate and quantify the system-level impact of gene variation.

  19. 27 CFR 24.95 - General.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false General. 24.95 Section 24.95 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE... wine spirits may be used for testing purposes, and wine may be used for tasting or sampling on bonded...

  20. 24 CFR 886.336 - Audit.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Audit. 886.336 Section 886.336... the Disposition of HUD-Owned Projects § 886.336 Audit. (a) Where a State or local government is the eligible owner of a project receiving financial assistance under this part, the audit requirements in 24...

  1. 24 CFR 883.313 - Audit.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Audit. 883.313 Section 883.313... § 883.313 Audit. (a) Where housing assistance under the Section 8 Program is provided for projects developed by State agencies, these agencies shall follow audit requirements in 24 CFR part 44. (b) Where a...

  2. 24 CFR 963.1 - Purpose.

    Science.gov (United States)

    2010-04-01

    ... by this part is based on the established procurement procedures set forth in 24 CFR 85.36, with... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Purpose. 963.1 Section 963.1... HOUSING-CONTRACTING WITH RESIDENT-OWNED BUSINESSES General § 963.1 Purpose. The purpose of this part is to...

  3. Functional consequences of T-stem mutations in E. coli tRNA Thr UGU in vitro and in vivo

    DEFF Research Database (Denmark)

    Saks, Margaret E; Sanderson, Lee E; Choi, Daniel S

    2011-01-01

    The binding affinities between Escherichia coli EF-Tu and 34 single and double base-pair changes in the T stem of E. coli tRNAThrUGU were compared with similar data obtained previously for several aa-tRNAs binding to Thermus thermophilus EF-Tu. With a single exception, the two proteins bound to m...

  4. Chromosome 15q24 microdeletion syndrome

    Directory of Open Access Journals (Sweden)

    Magoulas Pilar L

    2012-01-01

    Full Text Available Abstract Chromosome 15q24 microdeletion syndrome is a recently described rare microdeletion syndrome that has been reported in 19 individuals. It is characterized by growth retardation, intellectual disability, and distinct facial features including long face with high anterior hairline, hypertelorism, epicanthal folds, downslanting palpebral fissures, sparse and broad medial eyebrows, broad and/or depressed nasal bridge, small mouth, long smooth philtrum, and full lower lip. Other common findings include skeletal and digital abnormalities, genital abnormalities in males, hypotonia, behavior problems, recurrent infections, and eye problems. Other less frequent findings include hearing loss, growth hormone deficiency, hernias, and obesity. Congenital malformations, while rare, can be severe and include structural brain anomalies, cardiovascular malformations, congenital diaphragmatic hernia, intestinal atresia, imperforate anus, and myelomeningocele. Karyotypes are typically normal, and the deletions were detected in these individuals by array comparative genomic hybridization (aCGH. The deletions range in size from 1.7-6.1 Mb and usually result from nonallelic homologous recombination (NAHR between paralogous low-copy repeats (LCRs. The majority of 15q24 deletions have breakpoints that localize to one of five LCR clusters labeled LCR15q24A, -B, -C, -D, and -E. The smallest region of overlap (SRO spans a 1.2 Mb region between LCR15q24B to LCR15q24C. There are several candidate genes within the SRO, including CYP11A1, SEMA7A, CPLX3, ARID3B, STRA6, SIN3A and CSK, that may predispose to many of the clinical features observed in individuals with 15q24 deletion syndrome. The deletion occurred as a de novo event in all of the individuals when parents were available for testing. Parental aCGH and/or FISH studies are recommended to provide accurate genetic counseling and guidance regarding prognosis, recurrence risk, and reproductive options. Management

  5. 39 CFR 2.4 - Seal.

    Science.gov (United States)

    2010-07-01

    ... 39 Postal Service 1 2010-07-01 2010-07-01 false Seal. 2.4 Section 2.4 Postal Service UNITED STATES... Secretary of State, and is required by 39 U.S.C. 207 to be judicially noticed. The Seal shall be in the... description of the Postal Service emblem is described at 39 CFR 221.7. ...

  6. 24 CFR 3288.3 - Definitions.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 5 2010-04-01 2010-04-01 false Definitions. 3288.3 Section 3288.3... “manufactured home” as defined in 24 CFR 3280.2. Manufactured Housing Consensus Committee or MHCC means the consensus committee established pursuant to section 604(a)(3) of the Act, 42 U.S.C. 5403(a)(3). Party or...

  7. 24 CFR 886.131 - Audit.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Audit. 886.131 Section 886.131... Projects With HUD-Insured and HUD-Held Mortgages § 886.131 Audit. (a) Where a State or local government is... assistance under this part, the audit requirements in 24 CFR part 44 shall apply. (b) Where a nonprofit...

  8. 24 CFR 880.211 - Audit.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Audit. 880.211 Section 880.211... § 880.211 Audit. (a) Where a State or local government is the eligible owner of a project or a contract administrator under § 880.505 receiving financial assistance under this part, the audit requirements in 24 CFR...

  9. Thiol ligand-induced transformation of Au38(SC2H4Ph)24 to Au36(SPh-t-Bu)24.

    Science.gov (United States)

    Zeng, Chenjie; Liu, Chunyan; Pei, Yong; Jin, Rongchao

    2013-07-23

    We report a disproportionation mechanism identified in the transformation of rod-like biicosahedral Au38(SCH2CH2Ph)24 to tetrahedral Au36(TBBT)24 nanoclusters. Time-dependent mass spectrometry and optical spectroscopy analyses unambiguously map out the detailed size-conversion pathway. The ligand exchange of Au38(SCH2CH2Ph)24 with bulkier 4-tert-butylbenzenethiol (TBBT) until a certain extent starts to trigger structural distortion of the initial biicosahedral Au38(SCH2CH2Ph)24 structure, leading to the release of two Au atoms and eventually the Au36(TBBT)24 nanocluster with a tetrahedral structure, in which process the number of ligands is interestingly preserved. The other product of the disproportionation process, i.e., Au40(TBBT)m+2(SCH2CH2Ph)24-m, was concurrently observed as an intermediate, which was the result of addition of two Au atoms and two TBBT ligands to Au38(TBBT)m(SCH2CH2Ph)24-m. The reaction kinetics on the Au38(SCH2CH2Ph)24 to Au36(TBBT)24 conversion process was also performed, and the activation energies of the structural distortion and disproportionation steps were estimated to be 76 and 94 kJ/mol, respectively. The optical absorption features of Au36(TBBT)24 are interpreted on the basis of density functional theory simulations.

  10. Linear and nonlinear viscous flow in two-dimensional fluids

    International Nuclear Information System (INIS)

    Gravina, D.; Ciccotti, G.; Holian, B.L.

    1995-01-01

    We report on molecular dynamics simulations of shear viscosity η of a dense two-dimensional fluid as a function of the shear rate γ. We find an analytic dependence of η on γ, and do not find any evidence whatsoever of divergence in the Green-Kubo (GK) value that would be caused by the well-known long-time tail for the shear-stress autocorrelation function, as predicted by the mode-coupling theory. In accordance with the linear response theory, the GK value of η agrees remarkably well with nonequilibrium values at small shear rates. (c) 1995 The American Physical Society

  11. Dicty_cDB: FC-AI24 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available FC (Link to library) FC-AI24 (Link to dictyBase) - - - - FC-AI24Z (Link to Original site) - - FC-AI...24Z 693 - - - - Show FC-AI24 Library FC (Link to library) Clone ID FC-AI24 (Link to dictyBas...e) Atlas ID - NBRP ID - dictyBase ID - Link to Contig - Original site URL http://dictycdb.biol.tsukuba.ac.jp/CSM/FC/FC-AI/FC-AI...24Q.Seq.d/ Representative seq. ID FC-AI24Z (Link to Original s...ite) Representative DNA sequence >FC-AI24 (FC-AI24Q) /CSM/FC/FC-AI/FC-AI24Q.Seq.d/ XXXXXXXXXXAAATTAGAAAACAAA

  12. Luna 24 regolith breccias: A possible source of the fine size material of the Luna 24 regolith

    Science.gov (United States)

    Rode, O. D.; Lindstrom, M. M.

    1994-01-01

    The regolith breccias from the Luna 24 core were analyzed. The Luna 24 regolith is a mixture of fine and coarse grain materials. The comparable analysis of the grain size distributions, the modal and chemical compositions of the breccias, and the regolith from the same levels show that the friable slightly litificated breccia with a friable fine grain matrix may be a source of fine grain material of the Luna 24 present day regolith.

  13. 40 CFR 73.22-73.24 - [Reserved

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 16 2010-07-01 2010-07-01 false [Reserved] 73.22-73.24 Section 73.22-73.24 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) SULFUR DIOXIDE ALLOWANCE SYSTEM Allowance Allocations §§ 73.22-73.24 [Reserved] ...

  14. 22 CFR 212.24 - Effect of noncompliance.

    Science.gov (United States)

    2010-04-01

    ... 22 Foreign Relations 1 2010-04-01 2010-04-01 false Effect of noncompliance. 212.24 Section 212.24 Foreign Relations AGENCY FOR INTERNATIONAL DEVELOPMENT PUBLIC INFORMATION Availability of Information for Public Inspection and Copying § 212.24 Effect of noncompliance. No final order, opinion, statement of...

  15. 24,25,28-Trihydroxyvitamin D2 and 24,25,26-trihydroxyvitamin D2: Novel metabolites of vitamin D2

    International Nuclear Information System (INIS)

    Reddy, G.S.; Tserng, K.

    1990-01-01

    Understanding of the inactivation pathways of 25-hydroxyvitamin D 2 and 24-hydroxyvitamin D 2 , the two physiologically significant monohydroxylated metabolites of vitamin D 2 , is of importance, especially during hypervitaminosis D 2 . At present, little information is available regarding the inactivation pathway of 25-hydroxyvitamin D 2 except its further metabolism into 24,25-dihydroxyvitamin D 2 . In our present study, the authors investigated the metabolic fate of 25-hydroxyvitamin D 2 in the isolated perfused rat kidney and demonstrated its conversion not only into 24,25-dihydroxyvitamin D 2 but also into two other new metabolites, namely, 24,25,28-trihydroxyvitamin D 2 and 24,25,26-trihydroxyvitamin D 2 . The structure identification of the new metabolites was established by the techniques of ultraviolet absorption spectrophotometry and mass spectrometry and by the characteristic nature of each new metabolite's susceptibility to sodium metaperiodate oxidation. In order to demonstrate the physiological significance of the two new trihydroxy metabolites of vitamin D 2 , induced hypervitaminosis D 2 in a rat using [3α- 3 H]vitamin D 2 and analyzed its plasma for the various [3α- 3 H]vitamin D 2 metabolites on two different high-pressure liquid chromatography systems. The results indicate that both 24,25,26-trihydroxyvitamin D 2 and 24,25,26-trihydroxyvitamin D 2 circulate in the vitamin D 2 intoxicated rat in significant amounts along with other previously identified monohydroxy and dihydroxy metabolites of vitamin D 2 , namely, 24-hydroxyvitamin D 2 , 25-hydroxyvitamin D 2 , and 24,25-dihydroxyvitamin D 2

  16. 15 CFR 24.44 - Termination for convenience.

    Science.gov (United States)

    2010-01-01

    ... 15 Commerce and Foreign Trade 1 2010-01-01 2010-01-01 false Termination for convenience. 24.44 Section 24.44 Commerce and Foreign Trade Office of the Secretary of Commerce UNIFORM ADMINISTRATIVE... Reports, Records, Retention, and Enforcement § 24.44 Termination for convenience. Except as provided in...

  17. 27 CFR 10.24 - Sales promotion contests.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Sales promotion contests. 10.24 Section 10.24 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS COMMERCIAL BRIBERY Commercial Bribery § 10.24 Sales promotion contests...

  18. 7 CFR 1.24 - Preservation of records.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 1 2010-01-01 2010-01-01 false Preservation of records. 1.24 Section 1.24 Agriculture Office of the Secretary of Agriculture ADMINISTRATIVE REGULATIONS Official Records § 1.24 Preservation of records. Agencies shall preserve all correspondence relating to the requests it receives under this...

  19. 43 CFR 24.3 - General jurisdictional principles.

    Science.gov (United States)

    2010-10-01

    ... 43 Public Lands: Interior 1 2010-10-01 2010-10-01 false General jurisdictional principles. 24.3 Section 24.3 Public Lands: Interior Office of the Secretary of the Interior DEPARTMENT OF THE INTERIOR FISH AND WILDLIFE POLICY: STATE-FEDERAL RELATIONSHIPS § 24.3 General jurisdictional principles. (a) In...

  20. 12 CFR 24.3 - Public welfare investments.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 1 2010-01-01 2010-01-01 false Public welfare investments. 24.3 Section 24.3... DEVELOPMENT ENTITIES, COMMUNITY DEVELOPMENT PROJECTS, AND OTHER PUBLIC WELFARE INVESTMENTS § 24.3 Public welfare investments. A national bank or national bank subsidiary may make an investment directly or...

  1. 24-h Efficacy of Glaucoma Treatment Options.

    Science.gov (United States)

    Konstas, Anastasios G P; Quaranta, Luciano; Bozkurt, Banu; Katsanos, Andreas; Garcia-Feijoo, Julian; Rossetti, Luca; Shaarawy, Tarek; Pfeiffer, Norbert; Miglior, Stefano

    2016-04-01

    Current management of glaucoma entails the medical, laser, or surgical reduction of intraocular pressure (IOP) to a predetermined level of target IOP, which is commensurate with either stability or delayed progression of visual loss. In the published literature, the hypothesis is often made that IOP control implies a single IOP measurement over time. Although the follow-up of glaucoma patients with single IOP measurements is quick and convenient, such measurements often do not adequately reflect the untreated IOP characteristics, or indeed the quality of treated IOP control during the 24-h cycle. Since glaucoma is a 24-h disease and the damaging effect of elevated IOP is continuous, it is logical that we should aim to understand the efficacy of all treatment options throughout the 24-h period. This article first reviews the concept and value of diurnal and 24-h IOP monitoring. It then critically evaluates selected available evidence on the 24-h efficacy of medical, laser and surgical therapy options. During the past decade several controlled trials have significantly enhanced our understanding on the 24-h efficacy of all glaucoma therapy options. Nevertheless, more long-term evidence is needed to better evaluate the 24-h efficacy of glaucoma therapy and the precise impact of IOP characteristics on glaucomatous progression and visual prognosis.

  2. 14 CFR Sec. 2-4 - Accounting period.

    Science.gov (United States)

    2010-01-01

    ... 14 Aeronautics and Space 4 2010-01-01 2010-01-01 false Accounting period. Sec. 2-4 Section 2-4... REGULATIONS UNIFORM SYSTEM OF ACCOUNTS AND REPORTS FOR LARGE CERTIFICATED AIR CARRIERS General Accounting Provisions Sec. 2-4 Accounting period. (a) The accounting year of each air carrier subject to this Uniform...

  3. 49 CFR 230.24 - Maximum allowable stress.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 4 2010-10-01 2010-10-01 false Maximum allowable stress. 230.24 Section 230.24... Allowable Stress § 230.24 Maximum allowable stress. (a) Maximum allowable stress value. The maximum allowable stress value on any component of a steam locomotive boiler shall not exceed 1/4 of the ultimate...

  4. 24 CFR 92.356 - Conflict of interest.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 1 2010-04-01 2010-04-01 false Conflict of interest. 92.356... Development HOME INVESTMENT PARTNERSHIPS PROGRAM Other Federal Requirements § 92.356 Conflict of interest. (a... subrecipients, the conflict of interest provisions in 24 CFR 85.36 and 24 CFR 84.42, respectively, apply. In all...

  5. 19 CFR 24.14 - Salable Customs forms.

    Science.gov (United States)

    2010-04-01

    ... 19 Customs Duties 1 2010-04-01 2010-04-01 false Salable Customs forms. 24.14 Section 24.14 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY CUSTOMS FINANCIAL AND ACCOUNTING PROCEDURE § 24.14 Salable Customs forms. (a) Customs forms for sale to...

  6. Removal of 2,4-Dichlorophenolyxacetic acid (2,4-D) herbicide in the aqueous phase using modified granular activated carbon.

    Science.gov (United States)

    Dehghani, Mansooreh; Nasseri, Simin; Karamimanesh, Mojtaba

    2014-01-10

    Low cost 2,4-Dichlorophenolyxacetic acid (2,4-D) widely used in controlling broad-leafed weeds is frequently detected in water resources. The main objectives of this research were focused on evaluating the feasibility of using granular activated carbon modified with acid to remove 2,4-D from aqueous phase, determining its removal efficiency and assessing the adsorption kinetics. The present study was conducted at bench-scale method. The influence of different pH (3-9), the effect of contact time (3-90 min), the amount of adsorbent (0.1-0.4 g), and herbicide initial concentration (0.5-3 ppm) on 2,4-D removal efficiency by the granular activated carbon were investigated. Based on the data obtained in the present study, pH of 3 and contact time of 60 min is optimal for 2,4-D removal. 2,4-D reduction rate increased rapidly by the addition of the adsorbent and decreased by herbicide initial concentration (63%). The percent of 2,4-D reduction were significantly enhanced by decreasing pH and increasing the contact time. The adsorption of 2,4-D onto the granular activated carbon conformed to Langmuir and Freundlich models, but was best fitted to type II Langmuir model (R2 = 0.999). The second order kinetics was the best for the adsorption of 2,4-D by modified granular activated carbon with R2 > 0.99. Regression analysis showed that all of the variables in the process have been statistically significant effect (p granular activated carbon modified with acid is an appropriate method for reducing the herbicide in the polluted water resources.

  7. 42 CFR 81.24 - Guidelines for leukemia.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 1 2010-10-01 2010-10-01 false Guidelines for leukemia. 81.24 Section 81.24 Public... Causation § 81.24 Guidelines for leukemia. (a) For claims involving leukemia, DOL will calculate one or more probability of causation estimates from up to three of the four alternate leukemia risk models included in...

  8. 27 CFR 24.267 - Losses in transit.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Losses in transit. 24.267 Section 24.267 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT OF THE TREASURY LIQUORS WINE Losses of Wine § 24.267 Losses in transit. Where the loss in transit of...

  9. 24 CFR 700.175 - Other Federal requirements.

    Science.gov (United States)

    2010-04-01

    ... requirements described in 24 CFR part 44 (OMB Circular A-128). (b) Conflict of interest. In addition to the conflict of interest requirements in OMB Circular A-87 and 24 CFR part 85, no person who is an employee..., and the implementing regulations at 24 CFR part 108; and (3) Racial and ethnic collection requirements...

  10. Secreted phosphoprotein 24 kD (Spp24) inhibits growth of human pancreatic cancer cells caused by BMP-2

    International Nuclear Information System (INIS)

    Li, Chen-Shuang; Tian, Haijun; Zou, Min; Zhao, Ke-Wei; Li, Yawei; Lao, Lifeng; Brochmann, Elsa J.; Duarte, M. Eugenia L.; Daubs, Michael D.; Zhou, Yan-Heng; Murray, Samuel S.; Wang, Jeffrey C.

    2015-01-01

    The emerging role of bone morphogenetic proteins (BMPs) in the initiation and progression of multiple cancers has drawn great attention in cancer research. In this study, we report that BMP-2 can promote the proliferation of the pancreatic tumor cell line, PANC-1. Secreted phosphoprotein 24 kD (Spp24), a BMP binding protein, did not affect the proliferation of the cells but promoted the apoptosis of the cells in vitro. In a xeneograft tumor model using PANC-1 cells, BMP-2 dramatically promoted tumor growth, while Spp24 not only abolished the effect of BMP-2, but also dramatically induced tumor shrinking when used alone. Activation of Smad1/5/8 participated in this process as demonstrated by immunohistochemical staining of phosphorylated Smad 1/5/8. We conclude that Spp24 can be developed into a therapeutic agent that could be employed in clinical situations where the inhibition of BMPs and related proteins is advantageous. - Highlights: • Spp24 effectively inhibited the in vivo tumor growth of PANC-1. • BMP-2 dramatically promoted tumor growth by promoting PANC-1 proliferation. • Spp24 abolished the tumor growth effect of BMP-2 by promoting PANC-1 apoptosis. • Spp24 may be a candidate as a therapeutic agent of pancreatic cancer.

  11. Secreted phosphoprotein 24 kD (Spp24) inhibits growth of human pancreatic cancer cells caused by BMP-2

    Energy Technology Data Exchange (ETDEWEB)

    Li, Chen-Shuang [Department of Orthodontics, Peking University School and Hospital of Stomatology, Beijing (China); Tian, Haijun, E-mail: haijuntianmd@gmail.com [Department of Orthopaedic Surgery, Changzheng Hospital, Second Military Medical University, Shanghai (China); Department of Orthopaedic Surgery, University of California, Los Angeles, Los Angeles, CA (United States); Department of Surgery, Bethune School of Medics, Shijiazhuang (China); Zou, Min [Department of Orthodontics, School and Hospital of Stomatology, Xi' an Jiaotong University, Xi' an (China); Zhao, Ke-Wei [Research Service, VA Greater Los Angeles Healthcare System, North Hills, CA (United States); Li, Yawei; Lao, Lifeng [Department of Orthopaedic Surgery, University of California, Los Angeles, Los Angeles, CA (United States); Brochmann, Elsa J. [Research Service, VA Greater Los Angeles Healthcare System, North Hills, CA (United States); Geriatric Research, Education and Clinical Center, VA Greater Los Angeles Healthcare System, North Hills, CA (United States); Department of Medicine, University of California, Los Angeles, Los Angeles, CA (United States); Duarte, M. Eugenia L. [National Institute of Traumatology and Orthopaedics, Rio de Janeiro (Brazil); Daubs, Michael D. [Division of Orthopaedic Surgery, Department of Surgery, University of Nevada School of Medicine, Las Vegas, NV (United States); Zhou, Yan-Heng, E-mail: yanhengzhou@vip.163.com [Department of Orthodontics, Peking University School and Hospital of Stomatology, Beijing (China); Murray, Samuel S. [Research Service, VA Greater Los Angeles Healthcare System, North Hills, CA (United States); Geriatric Research, Education and Clinical Center, VA Greater Los Angeles Healthcare System, North Hills, CA (United States); Department of Medicine, University of California, Los Angeles, Los Angeles, CA (United States); Wang, Jeffrey C. [Department of Orthopaedic Surgery, University of Southern California, Los Angeles, CA (United States)

    2015-10-16

    The emerging role of bone morphogenetic proteins (BMPs) in the initiation and progression of multiple cancers has drawn great attention in cancer research. In this study, we report that BMP-2 can promote the proliferation of the pancreatic tumor cell line, PANC-1. Secreted phosphoprotein 24 kD (Spp24), a BMP binding protein, did not affect the proliferation of the cells but promoted the apoptosis of the cells in vitro. In a xeneograft tumor model using PANC-1 cells, BMP-2 dramatically promoted tumor growth, while Spp24 not only abolished the effect of BMP-2, but also dramatically induced tumor shrinking when used alone. Activation of Smad1/5/8 participated in this process as demonstrated by immunohistochemical staining of phosphorylated Smad 1/5/8. We conclude that Spp24 can be developed into a therapeutic agent that could be employed in clinical situations where the inhibition of BMPs and related proteins is advantageous. - Highlights: • Spp24 effectively inhibited the in vivo tumor growth of PANC-1. • BMP-2 dramatically promoted tumor growth by promoting PANC-1 proliferation. • Spp24 abolished the tumor growth effect of BMP-2 by promoting PANC-1 apoptosis. • Spp24 may be a candidate as a therapeutic agent of pancreatic cancer.

  12. 27 CFR 24.181 - Use of sugar.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Use of sugar. 24.181... OF THE TREASURY LIQUORS WINE Production of Wine § 24.181 Use of sugar. Only sugar, as defined in § 24.10, may be used in the production of standard wine. The quantity of sugar used will be determined...

  13. 12 CFR 563e.24 - Service test.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Service test. 563e.24 Section 563e.24 Banks and... Assessing Performance § 563e.24 Service test. (a) Scope of test. The service test evaluates a savings... performance, a community development service provided by an affiliate of the savings association, if the...

  14. 25 CFR 215.24 - Books and accounts.

    Science.gov (United States)

    2010-04-01

    ... 25 Indians 1 2010-04-01 2010-04-01 false Books and accounts. 215.24 Section 215.24 Indians BUREAU... LEASES, QUAPAW AGENCY § 215.24 Books and accounts. (a) The lessee shall maintain books in which shall be... land. (b) An audit of the lessee's accounts and books shall be made semiannually, or at such other...

  15. Whole genome transcript profiling of drug induced steatosis in rats reveals a gene signature predictive of outcome.

    Directory of Open Access Journals (Sweden)

    Nishika Sahini

    Full Text Available Drug induced steatosis (DIS is characterised by excess triglyceride accumulation in the form of lipid droplets (LD in liver cells. To explore mechanisms underlying DIS we interrogated the publically available microarray data from the Japanese Toxicogenomics Project (TGP to study comprehensively whole genome gene expression changes in the liver of treated rats. For this purpose a total of 17 and 12 drugs which are diverse in molecular structure and mode of action were considered based on their ability to cause either steatosis or phospholipidosis, respectively, while 7 drugs served as negative controls. In our efforts we focused on 200 genes which are considered to be mechanistically relevant in the process of lipid droplet biogenesis in hepatocytes as recently published (Sahini and Borlak, 2014. Based on mechanistic considerations we identified 19 genes which displayed dose dependent responses while 10 genes showed time dependency. Importantly, the present study defined 9 genes (ANGPTL4, FABP7, FADS1, FGF21, GOT1, LDLR, GK, STAT3, and PKLR as signature genes to predict DIS. Moreover, cross tabulation revealed 9 genes to be regulated ≥10 times amongst the various conditions and included genes linked to glucose metabolism, lipid transport and lipogenesis as well as signalling events. Additionally, a comparison between drugs causing phospholipidosis and/or steatosis revealed 26 genes to be regulated in common including 4 signature genes to predict DIS (PKLR, GK, FABP7 and FADS1. Furthermore, a comparison between in vivo single dose (3, 6, 9 and 24 h and findings from rat hepatocyte studies (2 h, 8 h, 24 h identified 10 genes which are regulated in common and contained 2 DIS signature genes (FABP7, FGF21. Altogether, our studies provide comprehensive information on mechanistically linked gene expression changes of a range of drugs causing steatosis and phospholipidosis and encourage the screening of DIS signature genes at the preclinical stage.

  16. Gamma knife radiosurgery for ten or more brain metastases. Analysis of the whole brain irradiation doses

    International Nuclear Information System (INIS)

    Nakaya, Kotaro; Hori, Tomokatsu; Izawa, Masahiro; Yamamoto, Masaaki

    2002-01-01

    Gamma knife (GK) radiosurgery has recently been recognized as the most powerful treatment modality in managing patients with brain metastasis, be they radioresistant or not, solitary or multiple. Very recently, this treatment has been employed in patients with numerous brain metastases, even those with 10 or more lesions. However, cumulative irradiation doses to the whole brain, with such treatment, remain unknown. Since the Gamma Plan ver. 5.10 (ver. 5.30 is presently available, Leksell Gamma Plan) became available in November, 1998, 105 GK procedures have been performed at our two facilities, Tokyo Women's Medical University and Katsuta Hospital Mito Gamma House. The median lesion number was 17, ranging 10-43, and the median cumulative volume of all tumors was 8.72 cm 3 , ranging 0.41-81.41 cm 3 . The selected doses at the lesion periphery ranged 12-25 Gy, the median being 20 Gy. Based on these treatment protocols, the cumulative irradiation dose was computed. The median cumulative irradiation dose to the whole brain was 4.83, ranging 2.16-8.51 Gy: the median integrated dose to the whole brain was 6.2 J, ranging 2.16-11.9 J. The median brain volumes receiving ≥2, ≥5, ≥10, ≥15 and ≥20 Gy were 1105 (range: 410-1501), 309 (46-1247), 64 (13-282), 24 (2-77), and 8 (0-40) cm 3 , respectively. The cumulative whole brain irradiation doses for patients with numerous radiosurgical targets were considered not to exceed the threshold level of normal brain necrosis. (author)

  17. 16 CFR 2.4 - Investigational policy.

    Science.gov (United States)

    2010-01-01

    ... 16 Commercial Practices 1 2010-01-01 2010-01-01 false Investigational policy. 2.4 Section 2.4 Commercial Practices FEDERAL TRADE COMMISSION ORGANIZATION, PROCEDURES AND RULES OF PRACTICE NONADJUDICATIVE... compulsory processes provided for by law. [45 FR 36341, May 29, 1980] ...

  18. 47 CFR 24.103 - Construction requirements.

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 2 2010-10-01 2010-10-01 false Construction requirements. 24.103 Section 24.103 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES PERSONAL... formula developed or generally used by industry, provided that such formula is based on the technical...

  19. The Use of Cone Beam Computed Tomography for Image Guided Gamma Knife Stereotactic Radiosurgery: Initial Clinical Evaluation

    Energy Technology Data Exchange (ETDEWEB)

    Li, Winnie; Cho, Young-Bin [Radiation Medicine Program, Princess Margaret Cancer Centre, Toronto, Ontario (Canada); Department of Radiation Oncology, University of Toronto, Toronto, Ontario (Canada); Ansell, Steve [Radiation Medicine Program, Princess Margaret Cancer Centre, Toronto, Ontario (Canada); Laperriere, Normand; Ménard, Cynthia; Millar, Barbara-Ann [Radiation Medicine Program, Princess Margaret Cancer Centre, Toronto, Ontario (Canada); Department of Radiation Oncology, University of Toronto, Toronto, Ontario (Canada); Zadeh, Gelareh [Division of Neurosurgery, University of Toronto University Health Network, Toronto Western Hospital, Toronto, Ontario (Canada); Macfeeters-Hamilton Centre for Neuro-oncology, Ontario Cancer Institute, Toronto, Ontario (Canada); Kongkham, Paul; Bernstein, Mark [Division of Neurosurgery, University of Toronto University Health Network, Toronto Western Hospital, Toronto, Ontario (Canada); Jaffray, David A. [Radiation Medicine Program, Princess Margaret Cancer Centre, Toronto, Ontario (Canada); Department of Radiation Oncology, University of Toronto, Toronto, Ontario (Canada); Department of Medical Biophysics, University of Toronto, Toronto, Ontario (Canada); Chung, Caroline, E-mail: caroline.chung.md@gmail.com [Radiation Medicine Program, Princess Margaret Cancer Centre, Toronto, Ontario (Canada); Department of Radiation Oncology, University of Toronto, Toronto, Ontario (Canada)

    2016-09-01

    Purpose: The present study used cone beam computed tomography (CBCT) to measure the inter- and intrafraction uncertainties for intracranial stereotactic radiosurgery (SRS) using the Leksell Gamma Knife (GK). Methods and Materials: Using a novel CBCT system adapted to the GK radiosurgery treatment unit, CBCT images were acquired immediately before and after treatment for each treatment session within the context of a research ethics board–approved prospective clinical trial. Patients were immobilized in the Leksell coordinate frame (LCF) for both volumetric CBCT imaging and GK-SRS delivery. The relative displacement of the patient's skull to the stereotactic reference (interfraction motion) was measured for each CBCT scan. Differences between the pre- and post-treatment CBCT scans were used to determine the intrafraction motion. Results: We analyzed 20 pre- and 17 post-treatment CBCT scans in 20 LCF patients treated with SRS. The mean translational pretreatment setup error ± standard deviation in the left-right, anteroposterior, and craniocaudal directions was −0.19 ± 0.32, 0.06 ± 0.27, and −0.23 ± 0.2 mm, with a maximum of −0.74, −0.53, and −0.68 mm, respectively. After an average time between the pre- and post-treatment CBCT scans of 82 minutes (range 27-170), the mean intrafraction error ± standard deviation for the LCF was −0.03 ± 0.05, −0.03 ± 0.18, and −0.03 ± 0.12 mm in the left-right, anteroposterior, and craniocaudual direction, respectively. Conclusions: Using CBCT on a prototype image guided GK Perfexion unit, we were able to measure the inter- and intrafraction positional changes for GK-SRS using the invasive frame. In the era of image guided radiation therapy, the use of CBCT image guidance for both frame- and non–frame-based immobilization systems could serve as a useful quality assurance tool. Our preliminary measurements can guide the application of achievable thresholds for inter- and intrafraction

  20. The Use of Cone Beam Computed Tomography for Image Guided Gamma Knife Stereotactic Radiosurgery: Initial Clinical Evaluation

    International Nuclear Information System (INIS)

    Li, Winnie; Cho, Young-Bin; Ansell, Steve; Laperriere, Normand; Ménard, Cynthia; Millar, Barbara-Ann; Zadeh, Gelareh; Kongkham, Paul; Bernstein, Mark; Jaffray, David A.; Chung, Caroline

    2016-01-01

    Purpose: The present study used cone beam computed tomography (CBCT) to measure the inter- and intrafraction uncertainties for intracranial stereotactic radiosurgery (SRS) using the Leksell Gamma Knife (GK). Methods and Materials: Using a novel CBCT system adapted to the GK radiosurgery treatment unit, CBCT images were acquired immediately before and after treatment for each treatment session within the context of a research ethics board–approved prospective clinical trial. Patients were immobilized in the Leksell coordinate frame (LCF) for both volumetric CBCT imaging and GK-SRS delivery. The relative displacement of the patient's skull to the stereotactic reference (interfraction motion) was measured for each CBCT scan. Differences between the pre- and post-treatment CBCT scans were used to determine the intrafraction motion. Results: We analyzed 20 pre- and 17 post-treatment CBCT scans in 20 LCF patients treated with SRS. The mean translational pretreatment setup error ± standard deviation in the left-right, anteroposterior, and craniocaudal directions was −0.19 ± 0.32, 0.06 ± 0.27, and −0.23 ± 0.2 mm, with a maximum of −0.74, −0.53, and −0.68 mm, respectively. After an average time between the pre- and post-treatment CBCT scans of 82 minutes (range 27-170), the mean intrafraction error ± standard deviation for the LCF was −0.03 ± 0.05, −0.03 ± 0.18, and −0.03 ± 0.12 mm in the left-right, anteroposterior, and craniocaudual direction, respectively. Conclusions: Using CBCT on a prototype image guided GK Perfexion unit, we were able to measure the inter- and intrafraction positional changes for GK-SRS using the invasive frame. In the era of image guided radiation therapy, the use of CBCT image guidance for both frame- and non–frame-based immobilization systems could serve as a useful quality assurance tool. Our preliminary measurements can guide the application of achievable thresholds for inter- and intrafraction

  1. SU-E-T-366: Estimation of Whole Body Dose From Cranial Irradiation From C and Perfexion Series Gamma Knife Units

    Energy Technology Data Exchange (ETDEWEB)

    Srivastava, S [Indiana University Health Methodist Hospital, Indianapolis, IN (United States); Indiana University School of Medicine, Indianapolis, IN, University Hospitals Case Medical Center, Cleaveland, OH (United States); Andersen, A; Lulu, B; Das, I [Indiana University School of Medicine, Indianapolis, IN, University Hospitals Case Medical Center, Cleaveland, OH (United States); Cheng, C

    2015-06-15

    Purpose: The Leksell Gamma Knife (GK) B & C series contains 201 Cobalt-60 sources with a helmet. The new model, Perfexion uses 192 Cobalt-60 sources without a helmet; using IRIS system for collimation and stereotactic guidance to deliver SRS to brain tumors. Relative dose to extracranial organs at risk (OARs) is measured in phantom in this study for Perfexion and C-series GK. Materials & Methods: Measurements were performed in a Rando anthropomorphic phantom on both systems using a large ion chamber (Keithley-175) for each collimator. The Keithley-175 cc ion chamber was sandwiched between phantom slices at various locations in the phantom to correspond to different extracranial OARs (thyroid, heart, kidney, ovary and testis, etc.) The dose measurement was repeated with OSL detectors for each position and collimator. Results: A large variation is observed in the normalized dose between these two systems. The dose beyond the housing falls off exponentially for Perfexion. Dose beyond the C-series GK housing falls off exponentially from 0–20cm then remains relatively constant from 20–40cm and again falls off with distance but less rapidly. The variation of extracranial dose with distance for each collimator is found to be parallel to each other for both systems. Conclusion: Whole body dose is found to vary significantly between these systems. It is important to measure the extracranial dose, especially for young patients. It is estimated that dose falls off exponentially from the GK housing and is about 1% for large collimators at 75 cm. The dose is two-orders of magnitude smaller for the 4mm collimator. However, this small dose for patient may be significant radiologically.

  2. Genomic-Enabled Prediction Kernel Models with Random Intercepts for Multi-environment Trials

    Science.gov (United States)

    Cuevas, Jaime; Granato, Italo; Fritsche-Neto, Roberto; Montesinos-Lopez, Osval A.; Burgueño, Juan; Bandeira e Sousa, Massaine; Crossa, José

    2018-01-01

    In this study, we compared the prediction accuracy of the main genotypic effect model (MM) without G×E interactions, the multi-environment single variance G×E deviation model (MDs), and the multi-environment environment-specific variance G×E deviation model (MDe) where the random genetic effects of the lines are modeled with the markers (or pedigree). With the objective of further modeling the genetic residual of the lines, we incorporated the random intercepts of the lines (l) and generated another three models. Each of these 6 models were fitted with a linear kernel method (Genomic Best Linear Unbiased Predictor, GB) and a Gaussian Kernel (GK) method. We compared these 12 model-method combinations with another two multi-environment G×E interactions models with unstructured variance-covariances (MUC) using GB and GK kernels (4 model-method). Thus, we compared the genomic-enabled prediction accuracy of a total of 16 model-method combinations on two maize data sets with positive phenotypic correlations among environments, and on two wheat data sets with complex G×E that includes some negative and close to zero phenotypic correlations among environments. The two models (MDs and MDE with the random intercept of the lines and the GK method) were computationally efficient and gave high prediction accuracy in the two maize data sets. Regarding the more complex G×E wheat data sets, the prediction accuracy of the model-method combination with G×E, MDs and MDe, including the random intercepts of the lines with GK method had important savings in computing time as compared with the G×E interaction multi-environment models with unstructured variance-covariances but with lower genomic prediction accuracy. PMID:29476023

  3. Vestibular schwannomas: clinical results and quality of life after microsurgery or gamma knife radiosurgery.

    Science.gov (United States)

    Myrseth, Erling; Møller, Per; Pedersen, Paal-Henning; Vassbotn, Flemming S; Wentzel-Larsen, Tore; Lund-Johansen, Morten

    2005-05-01

    The aim of the present study was to evaluate the overall treatment efficacy (tumor control, facial nerve function, complications) and quality of life for patients treated primarily for unilateral vestibular schwannomas of 30 mm or less, either by microsurgery or by gamma knife (GK) radiosurgery. The results for the two treatment groups are compared with each other, with main emphasis on the long-term quality of life. This is a retrospective study of 189 consecutive patients, 86 treated by microsurgery and 103 by gamma knife. The mean observation time was 5.9 years. All patients had a magnetic resonance imaging scan and clinical evaluation performed toward the end of the study. To evaluate the quality of life, we used two standardized questionnaires, the Glasgow Benefit Inventory and Short-Form 36. The questionnaires were sent to the 168 living patients. The reply rate was 83.3%. A total of 79.8% of the patients in the microsurgery group and 94.8% of the GK patients had a good facial nerve function (House-Brackmann Grade 1-2). Hearing was usually lost after microsurgery, whereas the GK patients had preserved hearing, which often became reduced over the years after the treatment. The treatment efficacy, defined as no need for additional treatment, was similar for the two treatment modalities. Quality of life was reduced compared with normative data, being most reduced in the microsurgery group. Some of the quality of life questions showed an association with facial nerve function and sex. Posttreatment facial nerve function, hearing, complication rates, and quality of life were all significantly in favor of GK radiosurgery.

  4. A 15q24 microduplication, reciprocal to the recently described 15q24 microdeletion, in a boy sharing clinical features with 15q24 microdeletion syndrome patients

    DEFF Research Database (Denmark)

    Lund, A.B. Kiholm; Hove, H.D.; Kirchhoff, M.

    2008-01-01

    A 15q24 microduplication, reciprocal to the minimal critical region for the recently described 15q24 microdeletion syndrome, was found in a 2-year-old boy by 244k Agilent oligoarray CGH analysis. The boy had global developmental delay and dysmorphic facial features, digital and genital abnormalit...

  5. 10 CFR 850.24 - Exposure monitoring.

    Science.gov (United States)

    2010-01-01

    ... 10 Energy 4 2010-01-01 2010-01-01 false Exposure monitoring. 850.24 Section 850.24 Energy... Exposure monitoring. (a) General. The responsible employer must ensure that: (1) Exposure monitoring is... activities properly. (b) Initial monitoring. The responsible employer must perform initial monitoring in...

  6. 47 CFR 24.249 - Payment issues.

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 2 2010-10-01 2010-10-01 false Payment issues. 24.249 Section 24.249 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES PERSONAL COMMUNICATIONS... issues. (a) Timing. On the day that a PCS entity files its prior coordination notice (PCN) in accordance...

  7. TBC1D24 genotype–phenotype correlation

    Science.gov (United States)

    Balestrini, Simona; Milh, Mathieu; Castiglioni, Claudia; Lüthy, Kevin; Finelli, Mattea J.; Verstreken, Patrik; Cardon, Aaron; Stražišar, Barbara Gnidovec; Holder, J. Lloyd; Lesca, Gaetan; Mancardi, Maria M.; Poulat, Anne L.; Repetto, Gabriela M.; Banka, Siddharth; Bilo, Leonilda; Birkeland, Laura E.; Bosch, Friedrich; Brockmann, Knut; Cross, J. Helen; Doummar, Diane; Félix, Temis M.; Giuliano, Fabienne; Hori, Mutsuki; Hüning, Irina; Kayserili, Hulia; Kini, Usha; Lees, Melissa M.; Meenakshi, Girish; Mewasingh, Leena; Pagnamenta, Alistair T.; Peluso, Silvio; Mey, Antje; Rice, Gregory M.; Rosenfeld, Jill A.; Taylor, Jenny C.; Troester, Matthew M.; Stanley, Christine M.; Ville, Dorothee; Walkiewicz, Magdalena; Falace, Antonio; Fassio, Anna; Lemke, Johannes R.; Biskup, Saskia; Tardif, Jessica; Ajeawung, Norbert F.; Tolun, Aslihan; Corbett, Mark; Gecz, Jozef; Afawi, Zaid; Howell, Katherine B.; Oliver, Karen L.; Berkovic, Samuel F.; Scheffer, Ingrid E.; de Falco, Fabrizio A.; Oliver, Peter L.; Striano, Pasquale; Zara, Federico

    2016-01-01

    Objective: To evaluate the phenotypic spectrum associated with mutations in TBC1D24. Methods: We acquired new clinical, EEG, and neuroimaging data of 11 previously unreported and 37 published patients. TBC1D24 mutations, identified through various sequencing methods, can be found online (http://lovd.nl/TBC1D24). Results: Forty-eight patients were included (28 men, 20 women, average age 21 years) from 30 independent families. Eighteen patients (38%) had myoclonic epilepsies. The other patients carried diagnoses of focal (25%), multifocal (2%), generalized (4%), and unclassified epilepsy (6%), and early-onset epileptic encephalopathy (25%). Most patients had drug-resistant epilepsy. We detail EEG, neuroimaging, developmental, and cognitive features, treatment responsiveness, and physical examination. In silico evaluation revealed 7 different highly conserved motifs, with the most common pathogenic mutation located in the first. Neuronal outgrowth assays showed that some TBC1D24 mutations, associated with the most severe TBC1D24-associated disorders, are not necessarily the most disruptive to this gene function. Conclusions: TBC1D24-related epilepsy syndromes show marked phenotypic pleiotropy, with multisystem involvement and severity spectrum ranging from isolated deafness (not studied here), benign myoclonic epilepsy restricted to childhood with complete seizure control and normal intellect, to early-onset epileptic encephalopathy with severe developmental delay and early death. There is no distinct correlation with mutation type or location yet, but patterns are emerging. Given the phenotypic breadth observed, TBC1D24 mutation screening is indicated in a wide variety of epilepsies. A TBC1D24 consortium was formed to develop further research on this gene and its associated phenotypes. PMID:27281533

  8. Dicty_cDB: SSM746 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available lcy*dlikirikn*pkqnllntlremailhtlivikflklltwiktdvflqmnykngskk eelitqewflqevqivkfsw...ilfykvlkfgt*kkkkik*ifnffk*sy*lylf* gk Frame B: ---gr*yf*gitgswcfvfns*sgp*w*ihisrnc*vlcrrgkkkmlnkmlrllhwlmlk

  9. 2,4-dichlorophenoxyacetic acid degradation in soils

    International Nuclear Information System (INIS)

    Bellinck, Celine; Batistic, L.; Mayaudon, J.

    1979-01-01

    The mineralization of 2,4-D (carboxyl 14 C) applied at 10 ppm was beginning immediately after its application in the soil. At higher concentrations a lag period appeared. The quantity of residual 14 C in the soil after 100 days incubation time reached 8.30 and 90% for initial concentrations of 10, 50 and 500 ppm of 2,4-D. The biostable 14 C which was extractable by water was exclusively constituted of 2,4-D. The mineral amendments P-K and PK inhibited 2,4-D mineralization, while the combination CaMg and also those containing nitrogen activated it. When 500 ppm of 2,4-D were applied one observed the formation of a zymogenous flora which was very active in the decarboxylation of the herbicide but unable to cleave the aromatic ring. These microorganisms probably belong to the genus Arthobacter and Pseudomonas [fr

  10. 30 CFR 784.24 - Road systems.

    Science.gov (United States)

    2010-07-01

    ... 30 Mineral Resources 3 2010-07-01 2010-07-01 false Road systems. 784.24 Section 784.24 Mineral... MINING AND RECLAMATION OPERATIONS PERMITS AND COAL EXPLORATION SYSTEMS UNDER REGULATORY PROGRAMS... systems. (a) Plans and drawings. Each applicant for an underground coal mining and reclamation permit...

  11. 40 CFR 503.24 - Management practices.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 29 2010-07-01 2010-07-01 false Management practices. 503.24 Section... FOR THE USE OR DISPOSAL OF SEWAGE SLUDGE Surface Disposal § 503.24 Management practices. (a) Sewage... the permitting authority that through management practices public health and the environment are...

  12. 27 CFR 24.77 - Experimental wine.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Experimental wine. 24.77... OF THE TREASURY LIQUORS WINE Administrative and Miscellaneous Provisions Tax Exempt Wine § 24.77 Experimental wine. (a) General. Any scientific university, college of learning, or institution of scientific...

  13. Analytic solutions for Rayleigh-Taylor growth rates in smooth density gradients

    International Nuclear Information System (INIS)

    Munro, D.H.

    1988-01-01

    The growth rate of perturbations on the shell of a laser fusion target can be estimated as √gk , where g is the shell acceleration and k is the transverse wave number of the perturbation. This formula overestimates the growth rate, and should be modified for the effects of density gradients and/or ablation of the unstable interface. The density-gradient effect is explored here analytically. With the use of variational calculus to explore all possible density profiles, the growth rate is shown to exceed √gk/(1+kL) , where L is a typical density-gradient scale length. Density profiles actually exhibiting this minimum growth rate are found

  14. Distributed Sleep Scheduling in Wireless Sensor Networks via Fractional Domatic Partitioning

    Science.gov (United States)

    Schumacher, André; Haanpää, Harri

    We consider setting up sleep scheduling in sensor networks. We formulate the problem as an instance of the fractional domatic partition problem and obtain a distributed approximation algorithm by applying linear programming approximation techniques. Our algorithm is an application of the Garg-Könemann (GK) scheme that requires solving an instance of the minimum weight dominating set (MWDS) problem as a subroutine. Our two main contributions are a distributed implementation of the GK scheme for the sleep-scheduling problem and a novel asynchronous distributed algorithm for approximating MWDS based on a primal-dual analysis of Chvátal's set-cover algorithm. We evaluate our algorithm with ns2 simulations.

  15. Synthesis of (E-2,4-Dinitro-N-((2E,4E-4-phenyl-5-(pyrrolidin-1-ylpenta-2,4-dienylideneaniline

    Directory of Open Access Journals (Sweden)

    Mostafa Fesanghari

    2009-07-01

    Full Text Available (E-2,4-Dinitro-N-((2E,4E-4-phenyl-5-(pyrrolidin-1-ylpenta-2,4-dienylidene aniline dye was prepared in one pot by reaction of premade N-2,4-dinitrophenyl-3-phenylpyridinium chloride (DNPPC and pyrrolidine in absolute MeOH.

  16. In anticipation of cycle 24

    Science.gov (United States)

    Kunches, Joseph; van der Linden, Ronald; Lundstedt, Henrik

    Anxious eyes are watching for the beginnings of new solar cycle 24. The watchers are many; scientists, engineers, academics, forecasters, end users. Their needs however vary, from the attraction of a better understanding of our nearest star, to how its increase in luminosity will affect particular aspects such as satellite orbits or the strength of the episodic storms that are most likely to occur during the height of the cycle. The first sign, however fleeting, occurred in January, 2008, when an ephemeral new cycle spot was numbered by NOAA as 10981. It was gone quickly but did mark the possible beginning of the new cycle. There has been great debate among solar physicists on how strong cycle 24 will be; and when it will start, reach maximum, and decline. This talk will give the status of the efforts to predict cycle 24, and the most recent projections.

  17. 24 CFR 594.28 - Environmental reviews.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 3 2010-04-01 2010-04-01 false Environmental reviews. 594.28... Grant Amounts § 594.28 Environmental reviews. (a) For all proposed actions or activities that are not considered categorically excluded under 24 CFR 50.20, HUD will perform the appropriate environmental reviews...

  18. 27 CFR 24.282 - Multiple transfers.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Multiple transfers. 24.282 Section 24.282 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT... transfer record for all wine (including distilling material and vinegar stock) transferred by pipeline to...

  19. 27 CFR 24.320 - Chemical record.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Chemical record. 24.320... OF THE TREASURY LIQUORS WINE Records and Reports § 24.320 Chemical record. A proprietor who uses chemicals, preservatives, or other such materials shall maintain a record of the purchase, receipt and...

  20. 28 CFR 24.307 - Department review.

    Science.gov (United States)

    2010-07-01

    ... review. The decision of the adjudicative officer will be reviewed to the extent permitted by law by the... 28 Judicial Administration 1 2010-07-01 2010-07-01 false Department review. 24.307 Section 24.307 Judicial Administration DEPARTMENT OF JUSTICE IMPLEMENTATION OF THE EQUAL ACCESS TO JUSTICE ACT IN...

  1. Production of 2-deoxyribose 5-phosphate from fructose to demonstrate a potential of artificial bio-synthetic pathway using thermophilic enzymes.

    Science.gov (United States)

    Honda, Kohsuke; Maya, Shohei; Omasa, Takeshi; Hirota, Ryuichi; Kuroda, Akio; Ohtake, Hisao

    2010-08-02

    Six thermophilic enzymes from Thermus thermophilus were used to construct an 'artificial bio-synthetic pathway' for the production of 2-deoxyribose 5-phosphate from fructose. By a simple operation using six recombinant Escherichia coli strains producing the thermophilic enzymes, respectively, fructose was converted to 2-deoxyribose 5-phosphate with a molar yield of 55%. Copyright 2010 Elsevier B.V. All rights reserved.

  2. 27 CFR 24.243 - Filtering aids.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Filtering aids. 24.243... OF THE TREASURY LIQUORS WINE Storage, Treatment and Finishing of Wine § 24.243 Filtering aids. Inert fibers, pulps, earths, or similar materials, may be used as filtering aids in the cellar treatment and...

  3. 24 CFR 3280.7 - Excluded structures.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 5 2010-04-01 2010-04-01 false Excluded structures. 3280.7 Section... DEVELOPMENT MANUFACTURED HOME CONSTRUCTION AND SAFETY STANDARDS General § 3280.7 Excluded structures. Certain structures may be excluded from these Standards as modular homes under 24 CFR 3282.12. [52 FR 4581, Feb. 12...

  4. 2 CFR 215.24 - Program income.

    Science.gov (United States)

    2010-01-01

    ... 2 Grants and Agreements 1 2010-01-01 2010-01-01 false Program income. 215.24 Section 215.24 Grants... income. (a) Federal awarding agencies shall apply the standards set forth in this section in requiring recipient organizations to account for program income related to projects financed in whole or in part with...

  5. 36 CFR 1210.24 - Program income.

    Science.gov (United States)

    2010-07-01

    ... 36 Parks, Forests, and Public Property 3 2010-07-01 2010-07-01 false Program income. 1210.24... Management § 1210.24 Program income. (a) The NHPRC applies the standards set forth in this section in requiring recipient organizations to account for program income related to projects financed in whole or in...

  6. 40 CFR 30.24 - Program income.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 1 2010-07-01 2010-07-01 false Program income. 30.24 Section 30.24... income. (a) EPA shall apply the standards set forth in this section in requiring recipient organizations to account for program income related to projects financed in whole or in part with Federal funds. (b...

  7. 24 CFR 1003.503 - Program income.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Program income. 1003.503 Section....503 Program income. (a) Program income requirements for ICDBG grantees are set forth in 24 CFR 85.25, as modified by this section. (b) Program income means gross income received by the grantee or a...

  8. 27 CFR 24.211 - Formula required.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Formula required. 24.211 Section 24.211 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT... which it is to be made, except that no formula is required for distilling material or vinegar stock. The...

  9. 27 CFR 24.218 - Other wine.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Other wine. 24.218 Section... THE TREASURY LIQUORS WINE Production of Other Than Standard Wine § 24.218 Other wine. (a) General. Other than standard wine not included in other sections in this subpart are considered other wine. Those...

  10. 42 CFR 24.5 - Peer review.

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 1 2010-10-01 2010-10-01 false Peer review. 24.5 Section 24.5 Public Health PUBLIC....5 Peer review. An individual may not be considered for appointment into the SBRS unless his/her qualifications have been reviewed by a PHS peer review committee and the committee has recommended appointment to...

  11. 27 CFR 24.317 - Sugar record.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Sugar record. 24.317... OF THE TREASURY LIQUORS WINE Records and Reports § 24.317 Sugar record. A proprietor who receives, stores, or uses sugar shall maintain a record of receipt and use. The record will show the date of...

  12. 12 CFR 25.24 - Service test.

    Science.gov (United States)

    2010-01-01

    ... option, the OCC will consider, in its assessment of a bank's service performance, a community development... performance rating. The OCC rates a bank's service performance as provided in appendix A of this part. ... 12 Banks and Banking 1 2010-01-01 2010-01-01 false Service test. 25.24 Section 25.24 Banks and...

  13. 32 CFR 32.24 - Program income.

    Science.gov (United States)

    2010-07-01

    ... 32 National Defense 1 2010-07-01 2010-07-01 false Program income. 32.24 Section 32.24 National... income. (a) DoD Components shall apply the standards set forth in this section in requiring recipient organizations to account for program income related to projects financed in whole or in part with Federal funds...

  14. 27 CFR 24.292 - Exported wine.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Exported wine. 24.292... OF THE TREASURY LIQUORS WINE Removal, Return and Receipt of Wine Removals Without Payment of Tax § 24.292 Exported wine. (a) General. Wine may be removed from a bonded wine premises without payment of tax...

  15. 27 CFR 24.203 - Honey wine.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Honey wine. 24.203 Section... THE TREASURY LIQUORS WINE Production of Agricultural Wine § 24.203 Honey wine. (a) Subject to paragraph (b) of this section, a winemaker, in the production of wine from honey, may add the following: (1...

  16. DNR 24K Streams

    Data.gov (United States)

    Minnesota Department of Natural Resources — 1:24,000 scale streams captured from USGS seven and one-half minute quadrangle maps, with perennial vs. intermittent classification, and connectivity through lakes,...

  17. 40 CFR 305.24 - Default order.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 27 2010-07-01 2010-07-01 false Default order. 305.24 Section 305.24... Default order. (a) Default. A party may be found to be in default: after motion, upon failure of the... default on the basis of failure to appear at a hearing shall be made against the Claims Official unless...

  18. DNR 24K Lakes

    Data.gov (United States)

    Minnesota Department of Natural Resources — Medium scale lake polygons derived from the National Wetlands Inventory (NWI) polygons and MnDOT Basemap lake delineations. Integrated with the DNR 24K Streams...

  19. 37 CFR 360.24 - Compliance with statutory dates.

    Science.gov (United States)

    2010-07-01

    ... dates. 360.24 Section 360.24 Patents, Trademarks, and Copyrights COPYRIGHT ROYALTY BOARD, LIBRARY OF CONGRESS SUBMISSION OF ROYALTY CLAIMS FILING OF CLAIMS TO ROYALTY FEES COLLECTED UNDER COMPULSORY LICENSE Digital Audio Recording Devices and Media Royalty Claims § 360.24 Compliance with statutory dates. (a...

  20. 7 CFR 322.24 - Packaging of transit shipments.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 5 2010-01-01 2010-01-01 false Packaging of transit shipments. 322.24 Section 322.24 Agriculture Regulations of the Department of Agriculture (Continued) ANIMAL AND PLANT HEALTH INSPECTION... Restricted Organisms Through the United States § 322.24 Packaging of transit shipments. (a) Restricted...

  1. 23_134 - 140_Uba G.K., et al.,_HEPATOCURATIVE

    African Journals Online (AJOL)

    user pc

    boratory Technology, College of Science and Technology, Jigawa State Polytechn. Dutse. istry, Faculty ... ERCP indicates improved fine architecture of the liver and kid ... of C. Procera may have moderate hepatorenal curative effects w ocera ...

  2. 7 CFR 70.24 - Financial interest of graders.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Financial interest of graders. 70.24 Section 70.24 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards... Graders § 70.24 Financial interest of graders. Graders shall not render service on any product in which...

  3. Dicty_cDB: CHL388 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available pcesvrarwaykvpgsxiqnllgmkhgapnkhhiilxk Frame B: gk*iin*dvniqtikrnrkrfhc**k*iqsctkq*nlngncqefrsqepqsfqslkmklm l*iyy*mksqkvlkscala...fmlptlx--- ---nqqr*innklrc*hsnn*kkpkkisllmkinselhqamkp*wklsri*kprttefqs lkmklml*iyy*mksqkvlkscalavvplmksvsrk

  4. Ostujõu pariteedid ja tegelikud kulutused / Viktoria Trasanov

    Index Scriptorium Estoniae

    Trasanov, Viktoria

    1996-01-01

    Ostujõud: pariteedid ja vahetuskursid. Kulutuste klassifikatsioonid: ICP ja SNA. EKS ja GK agregeerimismeetod. Euroopa 1993.a. Võrdlusprogrammi (ECP'93) struktuur. Võrdlusprogrammi korraldamine Balti grupis

  5. A Cascade of Thermophilic Enzymes As an Approach to the Synthesis of Modified Nucleotides.

    Science.gov (United States)

    Esipov, R S; Abramchik, Yu A; Fateev, I V; Konstantinova, I D; Kostromina, M A; Muravyova, T I; Artemova, K G; Miroshnikov, A I

    2016-01-01

    We propose a new approach for the synthesis of biologically important nucleotides which includes a multi-enzymatic cascade conversion of D -pentoses into purine nucleotides. The approach exploits nucleic acid exchange enzymes from thermophilic microorganisms: ribokinase, phosphoribosylpyrophosphate synthetase, and adenine phosphoribosyltransferase. We cloned the ribokinase gene from Thermus sp . 2.9, as well as two different genes of phosphoribosylpyrophosphate synthetase (PRPP-synthetase) and the adenine phosphoribosyltransferase (APR-transferase) gene from Thermus thermophilus HB27 into the expression vectors, generated high-yield E. coli producer strains, developed methods for the purification of the enzymes, and investigated enzyme substrate specificity. The enzymes were used for the conversion of D -pentoses into 5-phosphates that were further converted into 5-phospho-α- D -pentofuranose 1-pyrophosphates by means of ribokinase and PRPP-synthetases. Target nucleotides were obtained through the condensation of the pyrophosphates with adenine and its derivatives in a reaction catalyzed by APR-transferase. 2-Chloro- and 2-fluoroadenosine monophosphates were synthesized from D -ribose and appropriate heterobases in one pot using a system of thermophilic enzymes in the presence of ATP, ribokinase, PRPP-synthetase, and APR-transferase.

  6. Efecto de la conductancia de potasio Kv3.1 en la tasa y frecuencia de disparo de un axón no mielinado

    Directory of Open Access Journals (Sweden)

    Oscar Emilio Hernández Bustos

    2011-01-01

    Full Text Available Objetivo: Determinar la relación entre la conductancia de potasio Kv3.1 y la tasa de disparo (Td de un modelo neuronal llamado neurona1 formado por un soma, un cuello y un axón no mielinado durante un estímulo de corriente de 10 ms de duración y a 40°C. Materiales y métodos: A partir del software libre NEURON se simuló la propagación de ráfagas de potenciales de acción a través de neurona1, variando la conductancia específica máxima de potasio Kv3.1 (GKv3.1 relativa a la conductancia específica máxima de potasio (GK estudiada por A.L. Hodgkin y A.F. Huxley en 1952, de tal forma que GKv3.1+GK=1.6S/ cm2. Resultados: En una estructura neuronal con las características biofísicas de neurona1, Td varía en forma sigmoidea para 0 ¿ GKv3.1/GK ¿ 0.455 y decae exponencialmente para 0.455 < GKv3.1/GK ¿ 15, respectivamente. Para el primer caso, Td aumenta 11 veces más que la frecuencia (f respecto del número de espigas en cada ráfaga. Conclusión: La observación de la conductancia de potasio del tipo Kv3.1 en algún tipo de neurona no implica necesariamente la propagación de ráfagas de alta tasa de disparo. Su efecto es más pronunciado (11 veces en la modulación de Td que en el aumento de f.

  7. Nonlinear gyrokinetics: a powerful tool for the description of microturbulence in magnetized plasmas

    International Nuclear Information System (INIS)

    Krommes, John A

    2010-01-01

    Gyrokinetics is the description of low-frequency dynamics in magnetized plasmas. In magnetic-confinement fusion, it provides the most fundamental basis for numerical simulations of microturbulence; there are astrophysical applications as well. In this tutorial, a sketch of the derivation of the novel dynamical system comprising the nonlinear gyrokinetic (GK) equation (GKE) and the coupled electrostatic GK Poisson equation will be given by using modern Lagrangian and Lie perturbation methods. No background in plasma physics is required in order to appreciate the logical development. The GKE describes the evolution of an ensemble of gyrocenters moving in a weakly inhomogeneous background magnetic field and in the presence of electromagnetic perturbations with wavelength of the order of the ion gyroradius. Gyrocenters move with effective drifts, which may be obtained by an averaging procedure that systematically, order by order, removes gyrophase dependence. To that end, the use of the Lagrangian differential one-form as well as the content and advantages of Lie perturbation theory will be explained. The electromagnetic fields follow via Maxwell's equations from the charge and current density of the particles. Particle and gyrocenter densities differ by an important polarization effect. That is calculated formally by a 'pull-back' (a concept from differential geometry) of the gyrocenter distribution to the laboratory coordinate system. A natural truncation then leads to the closed GK dynamical system. Important properties such as GK energy conservation and fluctuation noise will be mentioned briefly, as will the possibility (and difficulties) of deriving nonlinear gyrofluid equations suitable for rapid numerical solution-although it is probably best to directly simulate the GKE. By the end of the tutorial, students should appreciate the GKE as an extremely powerful tool and will be prepared for later lectures describing its applications to physical problems.

  8. Nonlinear Gyrokinetics: A Powerful Tool for the Description of Microturbulence in Magnetized Plasmas

    International Nuclear Information System (INIS)

    Krommes, John E.

    2010-01-01

    Gyrokinetics is the description of low-frequency dynamics in magnetized plasmas. In magnetic-confinement fusion, it provides the most fundamental basis for numerical simulations of microturbulence; there are astrophysical applications as well. In this tutorial, a sketch of the derivation of the novel dynamical system comprising the nonlinear gyrokinetic (GK) equation (GKE) and the coupled electrostatic GK Poisson equation will be given by using modern Lagrangian and Lie perturbation methods. No background in plasma physics is required in order to appreciate the logical development. The GKE describes the evolution of an ensemble of gyrocenters moving in a weakly inhomogeneous background magnetic field and in the presence of electromagnetic perturbations with wavelength of the order of the ion gyroradius. Gyrocenters move with effective drifts, which may be obtained by an averaging procedure that systematically, order by order, removes gyrophase dependence. To that end, the use of the Lagrangian differential one-form as well as the content and advantages of Lie perturbation theory will be explained. The electromagnetic fields follow via Maxwell's equations from the charge and current density of the particles. Particle and gyrocenter densities differ by an important polarization effect. That is calculated formally by a 'pull-back' (a concept from differential geometry) of the gyrocenter distribution to the laboratory coordinate system. A natural truncation then leads to the closed GK dynamical system. Important properties such as GK energy conservation and fluctuation noise will be mentioned briefly, as will the possibility (and diffculties) of deriving nonlinear gyro fluid equations suitable for rapid numerical solution - although it is probably best to directly simulate the GKE. By the end of the tutorial, students should appreciate the GKE as an extremely powerful tool and will be prepared for later lectures describing its applications to physical problems.

  9. A Simple and Efficient Methodology To Improve Geometric Accuracy in Gamma Knife Radiation Surgery: Implementation in Multiple Brain Metastases

    Energy Technology Data Exchange (ETDEWEB)

    Karaiskos, Pantelis, E-mail: pkaraisk@med.uoa.gr [Medical Physics Laboratory, Medical School, University of Athens (Greece); Gamma Knife Department, Hygeia Hospital, Athens (Greece); Moutsatsos, Argyris; Pappas, Eleftherios; Georgiou, Evangelos [Medical Physics Laboratory, Medical School, University of Athens (Greece); Roussakis, Arkadios [CT and MRI Department, Hygeia Hospital, Athens (Greece); Torrens, Michael [Gamma Knife Department, Hygeia Hospital, Athens (Greece); Seimenis, Ioannis [Medical Physics Laboratory, Medical School, Democritus University of Thrace, Alexandroupolis (Greece)

    2014-12-01

    Purpose: To propose, verify, and implement a simple and efficient methodology for the improvement of total geometric accuracy in multiple brain metastases gamma knife (GK) radiation surgery. Methods and Materials: The proposed methodology exploits the directional dependence of magnetic resonance imaging (MRI)-related spatial distortions stemming from background field inhomogeneities, also known as sequence-dependent distortions, with respect to the read-gradient polarity during MRI acquisition. First, an extra MRI pulse sequence is acquired with the same imaging parameters as those used for routine patient imaging, aside from a reversal in the read-gradient polarity. Then, “average” image data are compounded from data acquired from the 2 MRI sequences and are used for treatment planning purposes. The method was applied and verified in a polymer gel phantom irradiated with multiple shots in an extended region of the GK stereotactic space. Its clinical impact in dose delivery accuracy was assessed in 15 patients with a total of 96 relatively small (<2 cm) metastases treated with GK radiation surgery. Results: Phantom study results showed that use of average MR images eliminates the effect of sequence-dependent distortions, leading to a total spatial uncertainty of less than 0.3 mm, attributed mainly to gradient nonlinearities. In brain metastases patients, non-eliminated sequence-dependent distortions lead to target localization uncertainties of up to 1.3 mm (mean: 0.51 ± 0.37 mm) with respect to the corresponding target locations in the “average” MRI series. Due to these uncertainties, a considerable underdosage (5%-32% of the prescription dose) was found in 33% of the studied targets. Conclusions: The proposed methodology is simple and straightforward in its implementation. Regarding multiple brain metastases applications, the suggested approach may substantially improve total GK dose delivery accuracy in smaller, outlying targets.

  10. Mechanisms of polyphosphate glucokinase and polyphosphate kinase reactions

    International Nuclear Information System (INIS)

    Pepin, C.A.; Robinson, N.A.; Wood, H.G.

    1986-01-01

    Polyphosphate glucokinase [poly(P)GK] catalyzes the following reaction: poly(P)/sub n/ + glucose → poly(P)/sub n-1/ + G-6-P. With long chain poly(P) [n=750 to 400] the mechanism appeared to be processive, in which there is phosphorylation of glucose without release of intermediate sizes of the poly(P) until the chain is about 100, thereafter there were intermediate sizes formed apparently by a non-processive process. Poly(P) kinase catalyzes the following reaction: ATP + poly(P)/sub n/ ↔ ADP + poly(P)/sub n + 1/. Using short chain 32 [P] poly(P) as a primer and non-radioactive ATP, long chain poly(P) is formed processively. The resulting chain has a short length labeled with 32 [P] contributed by the primer on one end and the remainder is made up of unlabeled (P) from the ATP. The authors have used this 32 [P] poly(P) as a substrate with poly(P)GK. If the mechanism of the poly(P)GK were initially processive, there would be a 50% chance the phosphate would be utilized from the unlabeled end and 50% of the 32 [P] would remain in the shortened chain. However, all the 32 [P] was lost when 20% of the poly(P) was converted to G-6-P. In contrast, with poly(P) kinase, the % of poly(P) utilized was equivalent to the % of 32 [P] converted to ATP, which is consistent with a strictly processive mechanism. Even though the mechanism of poly(P)GK appeared to be processive with long chains, the on and off rate of poly(P) from the enzyme is such that there is random removal of (P) from both ends of the poly(P) chain

  11. In type 1 diabetics, high-dose biotin may compensate for low hepatic insulin exposure, promoting a more normal expression of glycolytic and gluconeogenic enyzymes and thereby aiding glycemic control.

    Science.gov (United States)

    McCarty, Mark F

    2016-10-01

    In type 1 diabetics, hepatic exposure to insulin is chronically subnormal even in the context of insulin therapy; as a result, expression of glycolytic enzymes is decreased, and that of gluconeogenic enzymes is enhanced, resulting in a physiologically inappropriate elevation of hepatic glucose output. Subnormal expression of glucokinase (GK) is of particular importance in this regard. Possible strategies for correcting this perturbation of hepatic enzyme expression include administration of small molecule allosteric activators of GK, as well as a procedure known as chronic intermittent intravenous insulin therapy (CIIIT); however, side effects accompany the use of GK activators, and CIIIT is time and labor intensive. Alternatively, administration of high-dose biotin has potential for modulating hepatic enzyme expression in a favorable way. Studies in rodents and in cultured hepatocytes demonstrate that, in the context of low insulin exposure, supra-physiological levels of biotin induce increased expression of GK while suppressing that of the key gluconeogenic enzyme phosphoenolpyruvate carboxykinase. These effects may be a downstream consequence of the fact that biotin down-regulates mRNA expression of FOXO1; insulin's antagonism of the activity of this transcription factor is largely responsible for its modulatory impact on hepatic glycolysis and gluconeogenesis. Hence, high-dose biotin may compensate for subnormal insulin exposure by suppressing FOXO1 levels. High-dose biotin also has the potential to oppose hepatic steatosis by down-regulating SREBP-1 expression. Two pilot trials of high-dose biotin (16 or 2mg per day) in type 1 diabetics have yielded promising results. There is also some reason to suspect that high-dose biotin could aid control of diabetic neuropathy and nephropathy via its stimulatory effect on cGMP production. Owing to the safety, good tolerance, moderate expense, and current availability of high-dose biotin, this strategy merits more

  12. Phosphinothricin Acetyltransferases Identified Using In Vivo, In Vitro, and Bioinformatic Analyses

    Science.gov (United States)

    VanDrisse, Chelsey M.; Hentchel, Kristy L.

    2016-01-01

    ABSTRACT Acetylation of small molecules is widespread in nature, and in some cases, cells use this process to detoxify harmful chemicals. Streptomyces species utilize a Gcn5 N-acetyltransferase (GNAT), known as Bar, to acetylate and detoxify a self-produced toxin, phosphinothricin (PPT), a glutamate analogue. Bar homologues, such as MddA from Salmonella enterica, acetylate methionine analogues such as methionine sulfoximine (MSX) and methionine sulfone (MSO), but not PPT, even though Bar homologues are annotated as PPT acetyltransferases. S. enterica was used as a heterologous host to determine whether or not putative PPT acetyltransferases from various sources could acetylate PPT, MSX, and MSO. In vitro and in vivo analyses identified substrates acetylated by putative PPT acetyltransferases from Deinococcus radiodurans (DR_1057 and DR_1182) and Geobacillus kaustophilus (GK0593 and GK2920). In vivo, synthesis of DR_1182, GK0593, and GK2920 blocked the inhibitory effects of PPT, MSX, and MSO. In contrast, DR_1057 did not detoxify any of the above substrates. Results of in vitro studies were consistent with the in vivo results. In addition, phylogenetic analyses were used to predict the functionality of annotated PPT acetyltransferases in Burkholderia xenovorans, Bacillus subtilis, Staphylococcus aureus, Acinetobacter baylyi, and Escherichia coli. IMPORTANCE The work reported here provides an example of the use of a heterologous system for the identification of enzyme function. Many members of this superfamily of proteins do not have a known function, or it has been annotated solely on the basis of sequence homology to previously characterized enzymes. The critical role of Gcn5 N-acetyltransferases (GNATs) in the modulation of central metabolic processes, and in controlling metabolic stress, necessitates approaches that can reveal their physiological role. The combination of in vivo, in vitro, and bioinformatics approaches reported here identified GNATs that can

  13. Interdependency between mechanical parameters and afferent nerve discharge in remodeled diabetic Goto-Kakizaki rat intestine

    Directory of Open Access Journals (Sweden)

    Zhao J

    2017-12-01

    Full Text Available Jingbo Zhao,1 Jian Yang,1 Donghua Liao,1 Hans Gregersen2 1Giome Academia, Department of Clinical Medicine, Aarhus University, Aarhus, Denmark; 2Giome Center, Department of Surgery, Chinese University of Hong Kong and Prince of Wales Hospital, Shatin, Hong Kong Background: Gastrointestinal disorders are very common in diabetic patients, but the pathogenesis is still not well understood. Peripheral afferent nerves may be involved due to the complex regulation of gastrointestinal function by the enteric nervous system. Objective: We aimed to characterize the stimulus–response function of afferent fibers innervating the jejunum in the Goto-Kakizaki (GK type 2 diabetic rat model. A key question is whether changes in afferent firing arise from remodeled tissue or from adaptive afferent processes. Design: Seven 32-week-old male GK rats and seven age-matched normal Wistar rats were studied. Firing from mesenteric afferent nerves was recorded in excised jejunal segments of seven GK rats and seven normal Wistar rats during ramp test, stress relaxation test, and creep test. The circumferential stress–strain, spike rate increase ratio (SRIR, and single unit firing rates were calculated for evaluation of interdependency of the mechanical stimulations and the afferent nerve discharge. Results: Elevated sensitivity to mechanical stimuli was found for diabetic nerve bundles and single unit activity (P<0.05. The stress relaxed less in the diabetic intestinal segment (P<0.05. Linear association between SRIR and the thickness of circumferential muscle layer was found at high stress levels as well as for SRIR and the glucose level. Conclusion: Altered viscoelastic properties and elevated mechanosensitivity were found in the GK rat intestine. The altered nerve signaling is related to muscle layer remodeling and glucose levels and may contribute to gastrointestinal symptoms experienced by diabetic patients. Keywords: afferents, spike rate, stress–strain, creep

  14. Dosimetric and Clinical Analysis of Spatial Distribution of the Radiation Dose in Gamma Knife Radiosurgery for Vestibular Schwannoma

    International Nuclear Information System (INIS)

    Massager, Nicolas; Lonneville, Sarah; Delbrouck, Carine; Benmebarek, Nadir; Desmedt, Françoise; Devriendt, Daniel

    2011-01-01

    Objectives: We investigated variations in the distribution of radiation dose inside (dose inhomogeneity) and outside (dose falloff) the target volume during Gamma Knife (GK) irradiation of vestibular schwannoma (VS). We analyzed the relationship between some parameters of dose distribution and the clinical and radiological outcome of patients. Methods and Materials: Data from dose plans of 203 patients treated for a vestibular schwannoma by GK C using same prescription dose (12 Gy at the 50% isodose) were collected. Four different dosimetric indexes were defined and calculated retrospectively in all plannings on the basis of dose–volume histograms: Paddick conformity index (PI), gradient index (GI), homogeneity index (HI), and unit isocenter (UI). The different measures related to distribution of the radiation dose were compared with hearing and tumor outcome of 203 patients with clinical and radiological follow-up of minimum 2 years. Results: Mean, median, SD, and ranges of the four indexes of dose distribution analyzed were calculated; large variations were found between dose plans. We found a high correlation between the target volume and PI, GI, and UI. No significant association was found between the indexes of dose distribution calculated in this study and tumor control, tumor volume shrinkage, hearing worsening, loss of functional hearing, or complete hearing loss at last follow-up. Conclusions: Parameters of distribution of the radiation dose during GK radiosurgery for VS can be highly variable between dose plans. The tumor and hearing outcome of patients treated is not significantly related to these global indexes of dose distribution inside and around target volume. In GK radiosurgery for VS, the outcome seems more to be influenced by local radiation dose delivered to specific structures or volumes than by global dose gradients.

  15. 32 CFR 634.24 - Traffic planning and codes.

    Science.gov (United States)

    2010-07-01

    ... 32 National Defense 4 2010-07-01 2010-07-01 true Traffic planning and codes. 634.24 Section 634.24 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) LAW ENFORCEMENT AND CRIMINAL INVESTIGATIONS MOTOR VEHICLE TRAFFIC SUPERVISION Traffic Supervision § 634.24 Traffic planning and codes. (a) Safe and efficient...

  16. 32 CFR 635.24 - Updating the COPS MPRS.

    Science.gov (United States)

    2010-07-01

    ... 32 National Defense 4 2010-07-01 2010-07-01 true Updating the COPS MPRS. 635.24 Section 635.24 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) LAW ENFORCEMENT AND CRIMINAL INVESTIGATIONS LAW ENFORCEMENT REPORTING Offense Reporting § 635.24 Updating the COPS MPRS. Installation Provost Marshals/Directors of...

  17. Removal of 2,4-Dichlorophenolyxacetic acid (2,4-D) herbicide in the aqueous phase using modified granular activated carbon

    OpenAIRE

    Dehghani, Mansooreh; Nasseri, Simin; Karamimanesh, Mojtaba

    2014-01-01

    Background Low cost 2,4-Dichlorophenolyxacetic acid (2,4-D) widely used in controlling broad-leafed weeds is frequently detected in water resources. The main objectives of this research were focused on evaluating the feasibility of using granular activated carbon modified with acid to remove 2,4-D from aqueous phase, determining its removal efficiency and assessing the adsorption kinetics. Results The present study was conducted at bench-scale method. The influence of different pH (3–9), the ...

  18. Biodegradation of 2,4-dinitrotoluene by different plant species.

    Science.gov (United States)

    Podlipná, Radka; Pospíšilová, Blanka; Vaněk, Tomáš

    2015-02-01

    Over the past century, rapid growth of population, mining and industrialization significantly contributed to extensive soil, air and water contamination. The 2,4-dinitrotoluene (2,4-DNT), used mostly as explosive, belongs to the hazardous xenobiotics. Soils and waters contaminated with 2,4-DNT may be cleaned by phytoremediation using suitable plant species. The ability of crop plants (hemp, flax, sunflower and mustard) to germinate and grow on soils contaminated with 2,4-DNT was compared. Stimulation of their growth was found at 0.252 mg/g 2,4-DNT. The lethal concentration for the growth for these species was around 1 mg/g. In hydropony, the above mentioned species were able to survive 200 mg/l 2,4-DNT, the concentration close to maximal solubility of 2,4-DNT in water. Metabolism of 2,4-DNT was tested using suspension culture of soapwort and reed. The degradation products 2-aminonitrotoluene and 4-aminonitrotoluene were found both in the medium and in the acetone extract of plant cells. The test showed that the toxicity of these metabolites was higher than the toxicity of the parent compound, but 2,4-diaminotoluene, the product of next reduction step, was less toxic in the concentration range tested (0-200 mg/l). Copyright © 2014. Published by Elsevier Inc.

  19. Your Child's Development: 2 Years (24 Months)

    Science.gov (United States)

    ... 2 Years (24 Months) Print en español El desarrollo de su hijo: 2 años (24 meses) Tired ... and circular scribbles feeds himself or herself well Social and Emotional Development plays alongside other children fears ...

  20. Two new 24-isopropenyl-lanostanoids from Tillandsia brachycaulos.

    Science.gov (United States)

    Cantillo-Ciau, Zulema; Mena-Rejón, Gonzalo J; Quintero-Mármol, Esther; Jiménez-Díaz, Antonio; Quijano, Leovigildo

    2003-01-01

    The leaves of Tillandsia brachycaulos afforded two novel tetracyclic triterpenoids identified as (24S)-24-isopropenyl-29-nor-5alpha-lanosta-7-en-3beta-ol (1) and (24S)-24-isopropenyl-29-nor-5alpha-lanosta-7-en-3-one (2), in addition to the known isopimaric acid (3) and chlorogenic acid (4). Their structures were elucidated on the basis of spectral analysis, including homo- and heteronuclear correlation NMR experiments (COSY, ROESY, HMQC and HMBC) and by comparison with data in the literature. The antimicrobial and antifungal activities were studied. The compounds did not show significant activity.

  1. Recombinant interleukin-24 lacks apoptosis-inducing properties in melanoma cells.

    Directory of Open Access Journals (Sweden)

    Stephanie Kreis

    Full Text Available IL-24, also known as melanoma differentiation antigen 7 (mda-7, is a member of the IL-10 family of cytokines and is mainly produced by Th(2 cells as well as by activated monocytes. Binding of IL-24 to either of its two possible heterodimeric receptors IL-20R1/IL-20R2 and IL-22R/IL-20R2 activates STAT3 and/or STAT1 in target tissues such as lung, testis, ovary, keratinocytes and skin. To date, the physiological properties of IL-24 are still not well understood but available data suggest that IL-24 affects epidermal functions by increasing proliferation of dermal cells. In stark contrast to its "normal" and physiological behaviour, IL-24 has been reported to selectively and efficiently kill a vast variety of cancer cells, especially melanoma cells, independent of receptor expression and Jak-STAT signalling. These intriguing properties have led to the development of adenovirally-expressed IL-24, which is currently being evaluated in clinical trials. Using three different methods, we have analysed a large panel of melanoma cell lines with respect to IL-24 and IL-24 receptor expression and found that none of the investigated cell lines expressed sufficient amounts of functional receptor pairs and therefore did not react to IL-24 stimulation with Jak/STAT activation. Results for three cell lines contrasted with previous studies, which reported presence of IL-24 receptors and activation of STAT3 following IL-24 stimulation. Furthermore, evaluating four different sources and modes of IL-24 administration (commercial recombinant IL-24, bacterially expressed GST-IL-24 fusion protein, IL-24 produced from transfected Hek cells, transiently over-expressed IL-24 no induction or increase in cell death was detected when compared to appropriate control treatments. Thus, we conclude that the cytokine IL-24 itself has no cancer-specific apoptosis-inducing properties in melanoma cells.

  2. A simple and efficient methodology to improve geometric accuracy in gamma knife radiation surgery: implementation in multiple brain metastases.

    Science.gov (United States)

    Karaiskos, Pantelis; Moutsatsos, Argyris; Pappas, Eleftherios; Georgiou, Evangelos; Roussakis, Arkadios; Torrens, Michael; Seimenis, Ioannis

    2014-12-01

    To propose, verify, and implement a simple and efficient methodology for the improvement of total geometric accuracy in multiple brain metastases gamma knife (GK) radiation surgery. The proposed methodology exploits the directional dependence of magnetic resonance imaging (MRI)-related spatial distortions stemming from background field inhomogeneities, also known as sequence-dependent distortions, with respect to the read-gradient polarity during MRI acquisition. First, an extra MRI pulse sequence is acquired with the same imaging parameters as those used for routine patient imaging, aside from a reversal in the read-gradient polarity. Then, "average" image data are compounded from data acquired from the 2 MRI sequences and are used for treatment planning purposes. The method was applied and verified in a polymer gel phantom irradiated with multiple shots in an extended region of the GK stereotactic space. Its clinical impact in dose delivery accuracy was assessed in 15 patients with a total of 96 relatively small (series. Due to these uncertainties, a considerable underdosage (5%-32% of the prescription dose) was found in 33% of the studied targets. The proposed methodology is simple and straightforward in its implementation. Regarding multiple brain metastases applications, the suggested approach may substantially improve total GK dose delivery accuracy in smaller, outlying targets. Copyright © 2014 Elsevier Inc. All rights reserved.

  3. 27 CFR 30.24 - Specific gravity hydrometers.

    Science.gov (United States)

    2010-04-01

    ... hydrometers. 30.24 Section 30.24 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE... hydrometers. (a) The specific gravity hydrometers furnished by proprietors to appropriate TTB officers shall... instruments. Such specific gravity hydrometers shall be of a precision grade, standardization temperature 60...

  4. 28 CFR 94.24 - Determination of award.

    Science.gov (United States)

    2010-07-01

    ....24 Judicial Administration DEPARTMENT OF JUSTICE (CONTINUED) CRIME VICTIM SERVICES International Terrorism Victim Expense Reimbursement Program Coverage § 94.24 Determination of award. After review of each... fact and conclusions of law supporting it. A copy of the determination shall be mailed to the claimant...

  5. ASA24 enables multiple automatically coded self-administered 24-hour recalls and food records

    Science.gov (United States)

    A freely available web-based tool for epidemiologic, interventional, behavioral, or clinical research from NCI that enables multiple automatically coded self-administered 24-hour recalls and food records.

  6. Kinetics of Hesperetin for Liver Fortification in gamma-Irradiated Mice

    International Nuclear Information System (INIS)

    Tawfik, S.S.

    2011-01-01

    Hesperetin (3',5,7-trihydroxy-4'-methoxyflavonone), the aglycone of the flavanone glycosides hesperidin, exerts pharmacological properties such as antioxidation, anti-inflammation, blood lipid and cholesterol lowering is effectively used as a supplemental agent in the treatment protocols of complementary settings. Four groups were prepared: Control group: received 0.5 ml normal saline for 7 days. Hesperetin group: Mice received 7 doses of hesperetin injections (100 mg/ kg body wt/ day). Irradiated group: Mice submitted to total body irradiation with 4 Gy gamma-rays. Protected group (Hesperetin plus irradiation): Mice received hesperetin for 7 days and then submitted to 4 Gy of gamma-rays. The mice were sacrificed at 24 h, 1 week and 2 weeks after the end of the experimental treatments. Irradiated mice exhibited significant hyperglycaemia and augmented hepatic glycogen after the first day and 1 week but significant hypoglycemia and reducing hepatic glycogen after 2 weeks. Also, they exhibited significant increased serum total cholesterol (TC) and triacylglycerols (TG) and decreased hepatic TC and TG after 1 and 2 weeks. This treatment also resulted in a significant dropped in hepatic glucokinase (GK), glucose-6-phosphatase (G6P) and phosphoenolpyruvate carboxykinase (PEPCK) activities after 1 and 2 weeks. Hesperetin injections modulated the serum glucose and hepatic glycogen, adjusted TC and TG in both serum and liver and ameliorated the lessening in hepatic GK, G6P and PEPCK. The attending results demonstrated that hesperetn treatment modulated the biochemical symptoms of radiation disorders in mice. In conclusion, administration of hesperetin may have a useful role in modulating oxidative stress induced by exposure to gamma-radiation by improving the natural antioxidant mechanism and fortification liver functions

  7. ORF Alignment: NC_003282 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available LKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNGQPVDAKEM 60 ... Query: 236 VGTMTE-KSLSRGERKRLAFACEILTDPPILFC...DEPTSGLDSFMSHQVIKALRQLTIEGK 294 ... +G ... K LS GERKRLAFA E LTDPP+L CDEPTSGLDSF +H V++ L++L+ +GK Sbjct: 121 IGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQVLKKLSQKGK 180 ...

  8. 40 CFR 145.24 - Attorney General's statement.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 22 2010-07-01 2010-07-01 false Attorney General's statement. 145.24... (CONTINUED) STATE UIC PROGRAM REQUIREMENTS State Program Submissions § 145.24 Attorney General's statement... State Attorney General (or the attorney for those State or interstate agencies which have independent...

  9. 18 CFR 806.24 - Standards for diversions.

    Science.gov (United States)

    2010-04-01

    ... related to the following factors: (i) Any adverse effects and cumulative adverse effects the project may... 18 Conservation of Power and Water Resources 2 2010-04-01 2010-04-01 false Standards for diversions. 806.24 Section 806.24 Conservation of Power and Water Resources SUSQUEHANNA RIVER BASIN...

  10. 20 CFR 606.24 - Application for avoidance.

    Science.gov (United States)

    2010-04-01

    ... respect to which a State requests avoidance of tax credit reduction. The Governor is required to notify... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Application for avoidance. 606.24 Section 606.24 Employees' Benefits EMPLOYMENT AND TRAINING ADMINISTRATION, DEPARTMENT OF LABOR TAX CREDITS UNDER...

  11. 24-Hour Academic Libraries: Adjusting to Change

    Science.gov (United States)

    Bowman, Adam C.

    2013-01-01

    The purpose of this study was to explore the adaptive measures that academic libraries perform when implementing and operating a 24-hour schedule. Five in-depth interviews were conducted with current managerial-level librarians at 24-hour academic libraries. The exploratory interviews revealed similar measures for security, budgeting, employee…

  12. 27 CFR 24.101 - Bonded wine premises.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Bonded wine premises. 24..., DEPARTMENT OF THE TREASURY LIQUORS WINE Establishment and Operations Premises and Operations § 24.101 Bonded wine premises. (a) General. A person desiring to conduct operations involving untaxpaid wine, including...

  13. 27 CFR 24.141 - Bonded wine warehouse.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Bonded wine warehouse. 24..., DEPARTMENT OF THE TREASURY LIQUORS WINE Establishment and Operations Permanent Discontinuance of Operations § 24.141 Bonded wine warehouse. Where all operations at a bonded wine warehouse are to be permanently...

  14. AN ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) METHOD FOR THE URINARY BIOMONITORING OF 2,4-DICHLOROPHRENOCYACETIC ACID (2,4-D)

    Science.gov (United States)

    An enzyme-linked immunosorbent assay (ELISA) method was developed to quantitatively measure 2,4-dichlorophenoyacetic acid (2,4-D) in human urine. Samples were diluted (1:5) with phosphate-buffered saline, 0.05% Tween 20, with 0.02% sodium azide, and analyzed by a 96-microwekk pl...

  15. 33 CFR 385.24 - Project Management Plans.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Project Management Plans. 385.24... Processes § 385.24 Project Management Plans. (a) General requirements. (1) The Corps of Engineers and the... agencies, develop a Project Management Plan prior to initiating activities on a project. (2) The Project...

  16. 27 CFR 24.212 - High fermentation wine.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false High fermentation wine. 24..., DEPARTMENT OF THE TREASURY LIQUORS WINE Production of Other Than Standard Wine § 24.212 High fermentation wine. High fermentation wine is wine made with the addition of sugar within the limitations prescribed...

  17. 49 CFR 24.103 - Criteria for appraisals.

    Science.gov (United States)

    2010-10-01

    ...-year sales history of the property. (See appendix A, § 24.103(a)(1).) (ii) All relevant and reliable..., § 24.103(a).) (iii) A description of comparable sales, including a description of all relevant physical..., signature, and certification of the appraiser. (b) Influence of the project on just compensation. The...

  18. 27 CFR 24.166 - Buildings or rooms.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Buildings or rooms. 24.166... OF THE TREASURY LIQUORS WINE Construction and Equipment § 24.166 Buildings or rooms. All buildings or... afford adequate protection to the revenue. Each building or room will be constructed of substantial...

  19. 49 CFR 24.102 - Basic acquisition policies.

    Science.gov (United States)

    2010-10-01

    ... valuation only if the offer to acquire the property is $10,000, or less. (See appendix A, § 24.102(n).) [70... Transportation Office of the Secretary of Transportation UNIFORM RELOCATION ASSISTANCE AND REAL PROPERTY ACQUISITION FOR FEDERAL AND FEDERALLY-ASSISTED PROGRAMS Real Property Acquisition § 24.102 Basic acquisition...

  20. 27 CFR 24.303 - Formula wine record.

    Science.gov (United States)

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Formula wine record. 24..., DEPARTMENT OF THE TREASURY LIQUORS WINE Records and Reports § 24.303 Formula wine record. A proprietor who produces beverage formula wine shall maintain records showing by transaction date the details of production...