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1

Locked nucleic acid (LNA): High affinity targeting of RNA for diagnostics and therapeutics  

DEFF Research Database (Denmark)

Locked nucleic acid (LNA) is a nucleic acid analogue containing one or more LNA nucleotide monomers with a bicyclic furanose unit locked in an RNA mimicking sugar conformation. This conformational restriction results in unprecedented hybridization affinity towards complementary single stranded RNA and thus, makes LNA uniquely suited for mimicking RNA structures and sequence specific targeting of RNA in vitro or in vivo. The focus of this paper is on LNAantisense, LNA-modified siRNA (siLNA), and detection and analysis of microRNAs by LNA-modified oligonucleotide probes.

2005-01-01

2

A model for the single stranded random coil form of polydeoxyadenylic acid from minimum energy conformations of the dimeric subunit.  

UK PubMed Central (United Kingdom)

The minimum energy conformations of dApdA have been examined for their suitability as buildings blocks of the single stranded coil form of polynucleotides. Calculations of the characteristic ratio C...Full Text Available

1978-09-01

3

A single-stranded gap in human immunodeficiency virus unintegrated linear DNA defined by a central copy of the polypurine tract.  

UK PubMed Central (United Kingdom)

The structure of unintegrated human immunodeficiency virus type 1 (HIV-1) DNA from acutely infected human lymphoid cells was analyzed by nuclease S1 cleavage. We observed a unique, discrete single-stranded...Full Text Available

1991-05-01

4

Identification and characterization of noncoding small RNAs in Streptococcus pneumoniae serotype 2 strain D39.  

Science.gov (United States)

We report a search for small RNAs (sRNAs) in the low-GC, gram-positive human pathogen Streptococcus pneumoniae. Based on bioinformatic analyses by Livny et al. (J. Livny, A. Brencic, S. Lory, and M. K. Waldor, Nucleic Acids Res. 34:3484-3493, 2006), we tested 40 candidates by Northern blotting and confirmed the expression of nine new and one previously reported (CcnA) sRNAs in strain D39. CcnA is one of five redundant sRNAs reported by Halfmann et al. (A. Halfmann, M. Kovacs, R. Hakenbeck, and R. Bruckner, Mol. Microbiol. 66:110-126, 2007) that are positively controlled by the CiaR response regulator. We characterized 3 of these 14 sRNAs: Spd-sr17 (144 nucleotides [nt]; decreased in stationary phase), Spd-sr37 (80 nt; strongly expressed in all growth phases), and CcnA (93 nt; induced by competence stimulatory peptide). Spd-sr17 and CcnA likely fold into structures containing single-stranded regions between hairpin structures, whereas Spd-sr37 ...

2010-01-01

6

Insights into Cdc13 dependent telomere length regulation  

UK PubMed Central (United Kingdom)

Cdc13 is a single stranded telomere binding protein that specifically localizes to the telomere ends of budding yeasts and is essential for cell viability. It caps the ends of chromosomes thus preventing...Full Text Available

7

A rapid and efficient method for region- and strand-specific mutagenesis of cloned DNA.  

UK PubMed Central (United Kingdom)

The single-stranded viral DNA of an M13 phage recombinant containing the early promoter region of SV40 was hybridized with linear, double-stranded replicative form DNA of a related M13 phage containing...Full Text Available

1982-01-01

8

Transient suppression of MLH1 allows effective single-nucleotide substitution by single-stranded DNA oligonucleotides  

British Library Electronic Table of Contents (United Kingdom)

Short synthetic single-stranded oligodeoxyribonucleotides (ssODNs) can be used to introduce subtle modifications into the genome of mouse embryonic stem cells (ESCs). We have previously shown that effective application of ssODN-mediated gene targeting in ESC requires (transient) suppression of DNA mismatch repair (MMR). However, whereas transient down-regulation of the mismatch recognition protein MSH2 allowed substitution of 3 or 4 nucleotides, 1 or 2 nucleotide substitutions were still suppressed. We now demonstrate that single- or dinucleotide substitution can effectively be achieved by transient down-regulation of the downstream MMR protein MLH1. By exploiting highly specific real-time PCR, we demonstrate the feasibility of substituting a single basepair in a non-selectable gene. Howev...

2011-01-01

9

Shaping up nucleic acid computation  

UK PubMed Central (United Kingdom)

Summary of recent advances (abstract)Nucleic acid-based nanotechnology has always been perceived as novel, but has begun to move from theoretical demonstrations to practical applications....Full Text Available

2010-08-01

10

RNA:DNA Ratio and Other Nucleic Acid Derived Indices in Marine Ecology  

UK PubMed Central (United Kingdom)

Some of most used indicators in marine ecology are nucleic acid-derived indices. They can be divided by target levels in three groups: 1) at the organism level as ecophysiologic indicators, indicators...Full Text Available

11

Los Alamos sequence analysis package for nucleic acids and proteins.  

UK PubMed Central (United Kingdom)

An interactive system for computer analysis of nucleic acid and protein sequences has been developed for the Los Alamos DNA Sequence Database. It provides a convenient way to search or verify various...Full Text Available

1982-01-11

12

High-density multiplex detection of nucleic acid sequences: oligonucleotide ligation assay and sequence-coded separation.  

UK PubMed Central (United Kingdom)

We describe a non-isotopic, semi-automated method for large-scale multiplex analysis of nucleic acid sequences, using the cystic fibrosis transmembrane regulator (CFTR) gene as an example. Products...Full Text Available

1994-10-25

13

Comparative Evaluation of Three Commercial Systems for Nucleic Acid Extraction from Urine Specimens  

UK PubMed Central (United Kingdom)

A nucleic acid extraction system that can handle small numbers of specimens with a short test turnaround time and short hands-on time is desirable for emergent testing. We performed a comparative validation...Full Text Available

2005-09-01

14

Organisms posses enzymes that function in the repair of DNA damaged by radiations, chemicals and metabolic events  

International Nuclear Information System (INIS)

This report briefly describes the studies on the mechanism of in vivo DNA repairing by the author in Research Reactor Institute, Kyoto Univ. for the past 30 years. First, the ability of UV radiation to induce transformation was investigated with viral DNA. The formation of thymine-thymine dimer was found harmful to organisms and such dimers were removable by UV-radiation at a low frequency. The mutability was determined in three different E.coli strains with mutator gene, mutT, mutS or mutL. The ability to excise 8-oxoguanin developed in primer DNA was deficient in mutT and miss-pairing left after DNA replication could not be recovered in mutL and mutS strains. Further, DNA repairing mechanism was investigated in other microorganisms; single-strand cleavage caused by exposure to BNCB radiation (boron-neutron-captured beam) could not be repaired in E. coli. Whereas for Deinococcus radiodurans, of which survival rate was not decreased by #gamma#-ray radiation at 5 ...

1998-01-01

15

Comparison of the NucliSens easyMAG and Qiagen BioRobot 9604 Nucleic Acid Extraction Systems for Detection of RNA and DNA Respiratory Viruses in Nasopharyngeal Aspirate Samples?  

UK PubMed Central (United Kingdom)

The NucliSens easyMAG and BioRot 9604 automated nucleic acid extraction systems were evaluated and compared with the manual QIAamp (Qiagen) extraction method for their abilities to extract nucleic acid...Full Text Available

2008-07-01

16

The role of lesions of DNA in senescence of seeds of Lupinus polyphyllus L. induced by chronic low-intensity irradiation  

International Nuclear Information System (INIS)

A nonlinear relationship between the time of accelerated aging of Lupine seeds and the indices of its survival as well as the single-strand DNA amount in cells from these seeds is established. The character of this relationship is essentially altered in chronically irradiated lupine seeds from the Chernobyl NPP exclusion zone and seems more complicated. The possible role of repair systems in these effects is discussed. The fact that chronic irradiation in low doses can modify the course of senescence in lupine seeds reflects its high biological efficiency comparing with acute irradiation.

2000-08-01

17

Dipyrone-induced changes in DNA repair and other cell membrane associated processes in Escherichia coli  

Energy Technology Data Exchange (ETDEWEB)

The analgesic, dipyrone (1,phenyl-2,3-dimethyl-5-pyrazolone-4-methylamino methane sulphonate sodium), at 20 mM concentration, inhibited the rejoining of single-strand scissions in DNA of Escherichia coli B/r cells induced by 20 krad gamma-radiation. The chemical altered the cell membrane structure as evidenced from the uptake of acriflavin, the efflux of potassium ions from the bacterial cells and the inhibition of alkaline phosphatase-a cell membrane associated enzyme. (author). 18 refs., 6 figures.

1982-09-01

18

Dipyrone-induced changes in DNA repair and other cell membrane associated processes in Escherichia coli  

International Nuclear Information System (INIS)

The analgesic, dipyrone (1,phenyl-2,3-dimethyl-5-pyrazolone-4-methylamino methane sulphonate sodium), at 20 mM concentration, inhibited the rejoining of single-strand scissions in DNA of Escherichia coli B/r cells induced by 20 krad gamma-radiation. The chemical altered the cell membrane structure as evidenced from the uptake of acriflavin, the efflux of potassium ions from the bacterial cells and the inhibition of alkaline phosphatase-a cell membrane associated enzyme. (author).

19

Elastic properties of alternative versus single-stranded leveling archwires.  

Science.gov (United States)

The strength, stiffness, and range of single-stranded stainless steel (SS) and superelastic nickel-titanium (NiTi) archwires were compared with those of alternative leveling products, including nylon-coated and multistranded wires. Wire cross-sections were photographed after being potted in polymer, ground, and polished. Because the rectangular wires had rounded or beveled corners, gravimetric measurements and specific gravity calculations quantified the actual polygonal cross-sectional areas versus the ideal rectangular cross-sectional areas. Beveling reduced the cross-sectional areas by 7% to 8%; this decreased the wire stiffnesses by 15% to 19%. Using a testing machine, we measured the yield strengths, the elastic limits, and the ultimate tensile strengths in tension, and wire stiffnesses in 3-point bending. From cyclic loading tests, the elastic limits of the superelastic NiTi wires were approximately 90% and 45% of their ultimate tensile strengths for the ...

2002-11-01

20

Interactive computer programs in sequence data analysis.  

UK PubMed Central (United Kingdom)

We present interactive computer programs for the analysis of nucleic acid sequences. In order to handle these programs, minimum computer experience is sufficient. The nucleotide sequence of the human...Full Text Available

1982-01-11

 
 
 
 
21

Ultrafast fluorescence dynamics of Sybr Green I/DNA complexes  

British Library Electronic Table of Contents (United Kingdom)

The ultrafast dynamics of the DNA fluorescent dye Sybr Green I (SG) has been studied in buffer, single-stranded (ssDNA), double-stranded (dsDNA) and triple-stranded DNA (tsDNA). The fluorescence quantum yield of SG increases dramatically when bound to DNA (including tsDNA). The fluorescence dynamics of the free SG has shown two decay components with 0.15-0.4ps and 1.3-2.1ps time constants, depending on the fluorescence wavelength. Upon binding to DNA, the dynamics becomes slower exhibiting four decay components. This is mainly due to the restriction of the internal motions of the dye caused by the relatively rigid environment of the dye complexed with DNA.

2010-01-01

22

Mapping of the human cone transducin {alpha}-subunit (GNAT2) gene to 1p13 and negative mutation analysis in patients with Stargardt disease  

Energy Technology Data Exchange (ETDEWEB)

We report localization of the human cone transducin (GNAT2) gene using fluorescence in situ hybridization on chromosome 1 in band p13. The recent assignment of a gene for Stargardt disease to the same chromosomal region by linkage analysis prompted us to investigate the possible role of GNAT2 in the pathogenesis of this disease. We investigated 66 unrelated patients for mutations in the coding region of the GNAT2 gene using polymerase chain reaction-single strand conformation polymorphism analysis (SSCP) and direct sequencing. No disease-specific mutations were found, indicating that GNAT2 is probably not involved in the pathogenesis of most cases of Stargardt disease. 19 refs., 1 fig., 1 tab.

1995-01-01

23

Investigation by physical methods of the possible role of telomeres in DNA in aging process  

CERN Document Server

The interaction energies between the different types of bases of a single strand of DNA molecule have been calculated. Using these original values of energies the harmonic behavior of a number of base patterns of DNA has been studied. In view of the great interest aroused by the discovery of the role of the telomere segment of the DNA in the replication process and its possible link with the aging process, we have investigated, with simple models, the harmonic behavior of the telomeric pattern of bases as well as the thermodynamic response in the biological system. With these results a conclusion on the probable role of the telomeric pattern on aging has also been drawn. Here the calculated values of harmonic frequencies of the telomeric pattern of bases and of other possible patterns show that the telomeric pattern is associated with the highest vibrational frequency among all patterns of base combinations at the replication end of DNA. This ...

2003-01-01

24

Important effects of neighbouring nucleotides on electron induced DNA single-strand breaks  

British Library Electronic Table of Contents (United Kingdom)

In this Letter, we present Quantum Mechanics/Molecular Mechanics (QM/MM) calculations on molecules containing a 2-deoxycytidine-3prime-monophosphate moiety (3prime-dCMPH). In particular, we examine the effect that including neighbouring nucleotides at the Molecular Mechanic (MM) level has on the calculated electron affinities and on the energetic barriers of the C3prime-O3prime bond cleavage. Our results demonstrate that the surrounding nucleotides relocate the excess electron from the p* orbital of the base to a diffuse phosphate-centred orbital, leading to the formation of a dipole-bound anion state. Both the electron affinities and the activation energy of C3prime-O3prime bond cleavage are strongly increased.

2009-01-01

25

Effect of Diaporthe RNA virus 1 (DRV1) on growth and pathogenicity of different Diaporthe species  

British Library Electronic Table of Contents (United Kingdom)

A 4.1?kbp positive-strand RNA virus known as Diaporthe RNA virus 1 (DRV1) occurs in hypovirulent, non-sporulating isolates of the fungal pathogen Diaporthe perjuncta. A full-length cDNA clone of DRV1 was developed and RNA transcribed from the cDNA clone used to transfect different Diaporthe spp. The transfected species included three D. ambigua isolates and an unidentified Phomopsis asexual state of a Diaporthe sp. Successful transfections were confirmed using RT-PCR. Although the in vitro-transcribed positive sense single-stranded RNA used for transfection included vector sequences at both ends, the genomes of progeny virus from DRV1-transfected isolates were free of the vector sequences. Transfection resulted in morphological changes in these fungal pathogens. However, the presence of DR...

2011-01-01

26

Bioinformatic analysis of BBTV satellite DNA in Hainan  

British Library Electronic Table of Contents (United Kingdom)

Banana bunchy top virus (BBTV), family Nanaviridae, genus Babuvirus, is a single stranded DNA virus (ssDNA) that causes banana bunchy top disease (BBTD) in banana plants. It is the most common and most destructive of all viruses in these plants and is widespread throughout the Asia-Pacific region. In this study we isolated, cloned and sequenced a BBTV sample from Hainan Island, China. The results from sequencing and bioinformatics analysis indicate this isolate represents a satellite DNA component with 12 DNA sequences motifs. We also predicted the physical and chemical properties, structure, signal peptide, phosphorylation, secondary structure, tertiary structure and functional domains of its encoding protein, and compare them with the corresponding quantities in the replication initiatio...

2011-01-01

27

A connecting coupling  

Energy Technology Data Exchange (ETDEWEB)

The flexible, insulated, single strand cables are electrically connected with a cylindrical polar tip (PN) by means of cylindrical and conical shafts for the polar tips, which enter the faces of the divided, multiwire strand, clamped by tension half couplings. The flat ends of the polar tips being joined are positioned in two concentric mandrel bushings, an internal fixed one and an external, axially movable bushing (PV). The internal bushing is rigidly attached to the end of the left polar tip and equipped with three or four rounded, radial openings, whose diameter is determined by the external diameter of the locking device hinges (ShF) of the connecting couplings. The right polar tip is equipped with an annular channel of trapezoidal section into which the locking device hinges enter. The external movable bushing on the right side has a conical turning and on the left, a cylindrical into which the cylindrical spring which holds the movable ...

1981-06-05

28

Locked nucleic acid modified DNA enzymes targeting early growth response-1 inhibit human vascular smooth muscle cell growth  

UK PubMed Central (United Kingdom)

Smooth muscle cell (SMC) proliferation and migration are key processes that occur in the pathogenesis of atherosclerosis and post-angioplasty restenosis. In the present study, we designed locked nucleic...Full Text Available

2004-01-01

29

Nanocrystal Bioassembly: Asymmetry, Proximity, and Enzymatic Manipulation  

Energy Technology Data Exchange (ETDEWEB)

Research at the interface between biomolecules and inorganic nanocrystals has resulted in a great number of new discoveries. In part this arises from the synergistic duality of the system: biomolecules may act as self-assembly agents for organizing inorganic nanocrystals into functional materials; alternatively, nanocrystals may act as microscopic or spectroscopic labels for elucidating the behavior of complex biomolecular systems. However, success in either of these functions relies heavily uponthe ability to control the conjugation and assembly processes.In the work presented here, we first design a branched DNA scaffold which allows hybridization of DNA-nanocrystal monoconjugates to form discrete assemblies. Importantly, the asymmetry of the branched scaffold allows the formation of asymmetric2assemblies of nanocrystals. In the context of a self-assembled device, this can be considered a step toward the ability to engineer functionally distinct inputs and outputs.Next we develop an ...

2008-05-01

30

Comparative removal of pyrimidine dimers from human epidermal keratinocytes in vivo and in vitro  

Energy Technology Data Exchange (ETDEWEB)

We have compared the kinetics for repair of UV-induced cyclobutane pyrimidine dimers in the DNA of keratinocytes in human skin and in cell culture. A small area of the buttocks of volunteers was exposed to UVB-irradiation and biopsies were taken at various time intervals. Epidermal keratinocytes in culture from the same subjects were exposed to UVC with doses chosen to elicit comparable yields of dimers in cellular DNA. The initial density of pyrimidine dimers and the kinetics of their removal were assessed utilizing the dimer-specific T4 endonuclease V and sedimentation of the unlabeled DNA through alkaline sucrose gradients. The position of DNA in the gradients was determined using a monoclonal antibody against random sequences of single-stranded DNA in a sensitive immunoassay. The initial dimer frequency was 3.9-6.7 per 10(8) daltons DNA. About 40% of the dimers were removed within 1 h, 70% in 6 h, and 90% in 24 h for both in vivo and in vitro samples. The early ...

1988-10-01

31

Comparative removal of pyrimidine dimers from human epidermal keratinocytes in vivo and in vitro  

International Nuclear Information System (INIS)

We have compared the kinetics for repair of UV-induced cyclobutane pyrimidine dimers in the DNA of keratinocytes in human skin and in cell culture. A small area of the buttocks of volunteers was exposed to UVB-irradiation and biopsies were taken at various time intervals. Epidermal keratinocytes in culture from the same subjects were exposed to UVC with doses chosen to elicit comparable yields of dimers in cellular DNA. The initial density of pyrimidine dimers and the kinetics of their removal were assessed utilizing the dimer-specific T4 endonuclease V and sedimentation of the unlabeled DNA through alkaline sucrose gradients. The position of DNA in the gradients was determined using a monoclonal antibody against random sequences of single-stranded DNA in a sensitive immunoassay. The initial dimer frequency was 3.9-6.7 per 10(8) daltons DNA. About 40% of the dimers were removed within 1 h, 70% in 6 h, and 90% in 24 h for both in vivo and in vitro samples. The early ...

1988-01-01

32

Viral RNA testing and automation on the bead-based CBNE detection microsystem.  

Energy Technology Data Exchange (ETDEWEB)

We developed prototype chemistry for nucleic acid hybridization on our bead-based diagnostics platform and we established an automatable bead handling protocol capable of 50 part-per-billion (ppb) sensitivity. We are working towards a platform capable of parallel, rapid (10 minute), raw sample testing for orthogonal (in this case nucleic acid and immunoassays) identification of biological (and other) threats in a single sensor microsystem. In this LDRD we developed the nucleic acid chemistry required for nucleic acid hybridization. Our goal is to place a non-cell associated RNA virus (Bovine Viral Diarrhea, BVD) on the beads for raw sample testing. This key pre-requisite to showing orthogonality (nucleic acid measurements can be performed in parallel with immunoassay measurements). Orthogonal detection dramatically reduces false positives. We chose BVD because our collaborators ...

2008-09-01

33

Use of an alkaline phosphatase-labeled synthetic oligonucleotide probe for detection of Campylobacter jejuni and Campylobacter coli.  

UK PubMed Central (United Kingdom)

A commercially available synthetic nucleic acid probe (SNAP) conjugated to alkaline phosphatase was compared with standard culture techniques for detecting Campylobacter species. The SNAP was able to...Full Text Available

1990-07-01

34

Structural aberrations in group A Staphylococcus bacteriophages.  

UK PubMed Central (United Kingdom)

Six related Staphylococcus phages spontaneously produced various abnormal head and tail structures: (i) giant capsids which were tailed and apparently contained nucleic acid; (ii) regular and irregular...Full Text Available

1976-05-01

35

Reduced PCR Sensitivity Due to Impaired DNA Recovery with the MagNA Pure LC Total Nucleic Acid Isolation Kit  

UK PubMed Central (United Kingdom)

The increasing demand for molecular diagnostics in clinical microbiology laboratories necessitates automated sample processing. In the present study, we evaluated the performance of the MagNA Pure LC...Full Text Available

2005-09-01

36

Rational design of DNA sequences for nanotechnology, microarrays and molecular computers using Eulerian graphs  

UK PubMed Central (United Kingdom)

Nucleic acids are molecules of choice for both established and emerging nanoscale technologies. These technologies benefit from large functional densities of ‘DNA processing elements’...Full Text Available

2004-01-01

37

Rapid technique of DNA-DNA in situ hybridisation on formalin fixed tissue sections using microwave irradiation.  

UK PubMed Central (United Kingdom)

The relative sensitivities of different protocols for detecting cytomegalovirus nucleic acid sequences in histological specimens, using a biotinylated cDNA probe, were assessed. Several commonly used...Full Text Available

1987-08-01

38

GLUE-IT and PEDEL-AA: new programmes for analyzing protein diversity in randomized libraries  

UK PubMed Central (United Kingdom)

There are many methods for introducing random mutations into nucleic acid sequences. Previously, we described a suite of programmes for estimating the completeness and diversity of randomized DNA libraries...Full Text Available

2008-07-01

39

Does adenine incorporation into nucleic acids measure total microbial production:: a response to comments by Fuhrman et al  

Science.gov (United States)

The uptake and incorporation of (/sup 3/H)adenine as a measure of total microbial (bacteria and unicellular algae) nucleic acid synthesis depends on the validity of several assumptions. A unique characteristic of the (/sup 3/H)adenine method is the ability to measure the specific radioactivity (nCi pmol/sup -1/) of the immediate precursor pool of the adenine incorporated into nucleic acids. This measurement permits correction for isotope dilution when the added radiotracer mixes with exogenous pools of structurally related compounds before uptake and with endogenous pools after transport into the cells. The immediate precursors to (/sup 3/H)adenine incorporation into cellular RNA and DNA are ATP and dATP. Since the intracellular dATP pool is in isotopic equilibrium with the ATP pool, one can determine the specific radioactivity of both precursor pools by measuring that of the intracellular ATP pool. If the specific activity of the precursor ...

1986-11-01

40

Differential recognition of viral RNA by RIG-I  

UK PubMed Central (United Kingdom)

Retinoic acid inducible gene I (RIG-I) is a pattern recognition receptor (PRR) responsible for detection of nucleic acids from pathogens in the cytoplasm of infected cells and induction of type I interferon...Full Text Available

2011-03-01

 
 
 
 
41

Confocal microscopy for the analysis of siRNA delivery by polymeric nanoparticles  

UK PubMed Central (United Kingdom)

Clinical applications of genetic therapies, including delivery of short, interfering RNAs (siRNAs) for RNA interference (RNAi), are limited due to the difficulty of delivering nucleic acids...Full Text Available

2010-09-01

42

Comparative nucleic acid transfection efficacy in primary hepatocytes for gene silencing and functional studies  

UK PubMed Central (United Kingdom)

BackgroundPrimary hepatocytes are the best resource for in vitro studies directed at understanding hepatic processes at the cellular and molecular levels, necessary...Full Text Available

43

Comparative analysis of nitrifying bacteria associated with freshwater and marine aquaria.  

UK PubMed Central (United Kingdom)

Three nucleic acid probes, two for autotrophic ammonia-oxidizing bacteria of the beta subdivision of the class Proteobacteria and one for alpha subdivision nitrite-oxidizing bacteria, were developed...Full Text Available

1996-08-01

44

Being a binding site: Characterizing residue composition of binding sites on proteins  

UK PubMed Central (United Kingdom)

The Protein Data Bank contains the description of more than 45,000 three-dimensional protein and nucleic-acid structures today. Started to exist as the computer-readable depository of crystallographic...Full Text Available

45

Analyzing the simplicial decomposition of spatial protein structures  

UK PubMed Central (United Kingdom)

BackgroundThe fast growing Protein Data Bank contains the three-dimensional description of more than 45000 protein- and nucleic-acid structures today. The large majority of the data...Full Text Available

47

Evaluation of NucliSens easyMAG for automated nucleic acid extraction from various clinical specimens.  

Science.gov (United States)

The objectives of this study were to evaluate the performance of the NucliSens easyMAG platform for nucleic acid extraction from different clinical specimens compared to NucliSens miniMAG platform and manual QIAGEN extraction. The NucliSens easyMAG and the NucliSens miniMAG showed equal performance on 215 throat swabs since real-time nucleic acid sequence-based amplification scored the same samples positive for Mycoplasma pneumoniae (n=9) and Chlamydia pneumoniae (n=5) RNAs, although internal control RNA was slightly better detected with the NucliSens easyMAG (99.3% versus 96.8%). NucliSens easyMAG extracted nucleic acids more efficiently (higher recovery and/or fewer inhibitors) compared to QIAGEN extraction by showing, on average, lower Ct values in real-time LightCycler PCR, although 4 individual specimen out of 45 were found positive only with QIAGEN. For nine M. pneumoniae-positive throat swabs, the mean difference in ...

2006-12-13

48

Construction and Analytical Application of Internal Amplification Controls (IAC) for Detection of Food Supply Chain-Relevant Viruses by Real-Time PCR-Based Assays  

British Library Electronic Table of Contents (United Kingdom)

Internal amplification controls (IACs) were constructed for incorporation into real-time nucleic acid amplification assays for bovine polyomavirus, hepatitis A virus, hepatitis E virus, human adenovirus, human norovirus genogroup I, human norovirus genogroup II, murine norovirus and porcine adenovirus. The addition of optimised amounts of IAC into the assays did not affect the limits of detection for each specific target virus. A poorly performed extraction of viral nucleic acids was simulated, and the effectiveness of IACs in identifying failed assays was demonstrated. The IACs constructed in this study can be reliably used in their specific assays to provide a robust control that can be routinely applied in the analysis of foods for viruses.

2011-01-01

49

Comparison of fluorescence in situ hybridisation using peptide nucleic acid probes, Gram stain/acridine orange leukocyte cytospin and differential time to positivity methods for detection of catheter related bloodstream infection in patients after haematopoietic stem cell transplantation  

British Library Electronic Table of Contents (United Kingdom)

Abstract In 46 febrile neutropenic patients who had undergone haematopoietic stem cell transplantation, the fluorescence in situ hybridisation using peptide nucleic acid probes (PNA FISH), Gram stain/acridine orange leukocyte cytospin (Gram/AOLC), and differential time to positivity (DTP) methods were performed for detection of catheter related bloodstream infections (CRBSIs). As compared with the DTP method (which detected 11 patients with CRBSI), the PNA FISH and the Gram/AOLC methods detected ten of 11 CRBSI patients, resulting in a sensitivity, specificity, negative predictive value and positive predictive value of 91%, 100%, 97% and 100%, respectively, for the PNA FISH method as well as for the Gram/AOLC method.

2010-01-01

50

Theoretical studies of metal-phosphate interactions: interaction of Li+, Na+, K+, Be++, Mg++, and Ca++ with H2PO4- and (CH3O)2PO2-: implications for nucleic acid solvation.  

UK PubMed Central (United Kingdom)

Model phosphate-metal solvation complexes have been studied by ab-initio self-consistent-field techniques. The complexes studied include (RO)2PO2-(R = H or CH3) with Li+, Na+, K+, Be++, Mg++, Ca++,...Full Text Available

1975-10-01

51

The DNA and RNA specificity of eilatin Ru(II) complexes as compared to eilatin and ethidium bromide  

UK PubMed Central (United Kingdom)

Eilatin-containing ruthenium complexes bind to a broad range of different nucleic acids including: calf thymus (CT) DNA, tRNAPhe, polymeric RNAs and DNAs, and viral RNAs including the HIV-1...Full Text Available

2003-10-01

52

Ribose metabolism and nucleic acid synthesis in normal and glucose-6-phosphate dehydrogenase-deficient human erythrocytes infected with Plasmodium falciparum.  

UK PubMed Central (United Kingdom)

The metabolism of pentose-phosphate was investigated in Plasmodium falciparum-infected normal and glucose-6-phosphate dehydrogenase (G6PD)-deficient human red blood cells in vitro. 5'-Phosphoribosyl-1-pyrophosphate...Full Text Available

1986-04-01

53

Human thrombomodulin gene is intron depleted: nucleic acid sequences of the cDNA and gene predict protein structure and suggest sites of regulatory control.  

UK PubMed Central (United Kingdom)

We have isolated a human thrombomodulin cDNA, and a human genomic clone containing the putative promoter domain, as well as the translated and untranslated regions of the endothelial cell receptor....Full Text Available

1987-09-01

54

Very low-dose hyper-radiosensitivity: impact for radiotherapy of micrometastases; Hyperradiosensibilite aux tres faibles doses: impact en radiotherapie des micrometastases  

Energy Technology Data Exchange (ETDEWEB)

Radio-biologists have pointed out a novel radiobiological phenomenon observed in many tumor and normal cell lines: hyper-radiosensitivity to very low-dose (H.R.S.) followed by induced radioresistance (I.R.R.) after a threshold dose of 0.1-0.3 Gy that depends on the cell line. Radioresistance at high dose (i.e. higher than 0.5 Gy) and metastatic potential of tumor cells are likely major factors of failure in radiotherapy. A careful review of literature suggests that: 1) radiotherapy does not increase the metastatic potential of tumor cells; 2) radioresistance at high dose and metastatic potential are not related. However, inside a given tumor cell line, highly metastatic clones may elicit more cells showing H.R.S; or are more radiosensitive at high dose than poorly metastatic ones. Recent data obtained from molecular techniques (comet and immunofluorescence assays) applied to single cells irradiated at very low radiation doses (1-100 mGy) suggest that DNA ...

2007-09-15

55

Mapping of the human cone transducin {alpha} subunit (GNAT2) gene to 1p13 and mutation analysis in patients with Stargardt`s disease  

Energy Technology Data Exchange (ETDEWEB)

Transducin {alpha} subunits are members of a large family of G-proteins and play an important role in phototransduction in rod and cone photoreceptors. We report the localization of the human cone {alpha} transducin (GNAT2) gene using fluorescence in situ hybridization (FISH) on chromosome 1 in band p13. The recent assignment of a gene for Stargardt`s disease to the same chromosomal region by linkage analysis prompted us to investigate the possible role of GNAT2 in the pathogenesis of this disease. Stargardt`s disease is characterized by degeneration in late childhood or early adulthood of the macula of the retina, a region rich in cones. We screened patients with Stargardt`s disease, with or without peripheral cone involvement as monitored by the full-field ERG, for mutations in this gene. We investigated 66 unrelated patients including 22 with peripheral cone dysfunction for mutations in the coding region of the GNAT2 gene using polymerase chain reaction-single ...

1994-09-01

56

Elastic flexural properties of multistranded stainless steel versus conventional nickel titanium archwires.  

Science.gov (United States)

Based on a recent investigation that modeled the elastic properties (ie, strength, stiffness and range) of multistranded wires made from linearly elastic materials, three-stranded (triple) and six-stranded coaxial (coax) stainless steel (SS) wires were compared to single-stranded (single) SS and conventional nickel titanium (NiTi) leveling wires. To measure Young's modulus of elasticity (E), flexural tests were performed with an Instron mechanical testing machine in a three-point bending arrangement having a span length of 8.9 mm or 12.5 mm. A strong correlation between wire stiffness and the area moment of inertia demonstrated that strand interaction was negligible at low activations and that E = 199 GPa was constant even for the heavily drawn coax strands. Using the Instron with an extensometer, the 0.1% yield strengths (sigma(YS)) of the single SS wires and the straight inner strands within the coax wires were tested. The ratio of the sigma(YS) to the ultimate ...

2002-08-01

57

The wavelength dependence of ultraviolet enhanced reactivation in a mammalian cell-virus system  

International Nuclear Information System (INIS)

The effect of UV radiation in the wavelength region 230 nm to 302 nm on the ability of an irradiated mammalian cell to reactivate UV-irradiated mammalian virus was tested. An action spectrum for radiation enhanced reactivation (RER) is presented. The shape of the action spectrum points to a combined nucleic acid-protein target for UV radiation effects on this cellular parameter. An analysis of the results of others involving the biochemical and photobiological events involved in RER does not allow us to distinguish which macromolecule is the major contributor to this effect. Studies involving an analogous phenomenon in bacteris (Weigle reactivation) imply that RER and WR amy involve similar mechanisms. (author).

58

Magnetic silica extraction for low-viremia human immunodeficiency virus type 1 genotyping.  

Science.gov (United States)

Nucleic acid extraction and human immunodeficiency virus type 1 (HIV-1) genotyping using the NucliSens miniMAG platform and the TruGene HIV-1 genotyping kit gave HIV-1 sequence data from HIV-1-negative plasma spiked with 100 copies/ml reference HIV-1 RNA and from low-viremia clinical samples (<500 copies/ml) without the need for ultracentrifugation or nested second-round PCR. PMID:17122019

2006-11-22

59

Cysteine-containing peptides having antioxidant properties  

Energy Technology Data Exchange (ETDEWEB)

The term "homology" or "homologous" means an amino acid similarity measured by the program, BLAST (Altschul et al (1997), "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs", Nucleic Acids Res. 25:33 89 3402), and expressed as --(% identity n/n). In measuring homology between a peptide and a protein of greater size, homology is measured only in the corresponding region; that is, the protein is regarded as only having the same general length as the peptide, allowing for gaps and insertions.

2007-05-15

60

Platinum(II) complexes as spectroscopic probes for biomolecules  

Energy Technology Data Exchange (ETDEWEB)

The use of platinum(II) complexes as tags and probes for biomolecules is indeed advantageous for their reactivities can be selective for certain purposes through an interplay of mild reaction conditions and of the ligands bound to the platinum. The use of {sup 195}Pt NMR as a method of detecting platinum and its interactions with biomolecules was carried out with the simplest model of platinum(II) tagging to proteins. Variable-temperature {sup 195}Pt NMR spectroscopy proved useful in studying the stereodynamics of complex thioethers like methionine. The complex, Pt(trpy)Cl{sup +}, with its chromophore has a greater potential for probing proteins. It is a noninvasive and selective tag for histidine and cysteine residues on the surface of cytochrome c at pH 5. The protein derivatives obtained are separable, and the tags are easily quantitated and differentiated through the metal-to-ligand charge transfer bands which are sensitive to the environment of the tag. Increasing the pH to 7.0 ...

1990-09-21

 
 
 
 
61

Sequence analysis of the ATM gene in 20 patients with RTOG grade 3 or 4 acute and/or late tissue radiation side effects  

International Nuclear Information System (INIS)

Purpose: Patients with ataxia-telangiectasia (A-T) show greatly increased radiation sensitivity and cancer predisposition. Family studies imply that the otherwise clinically silent heterozygotes of this autosomal recessive disease run a 3.5 to 3.8 higher risk of developing cancer. In vitro studies suggest moderately increased cellular radiation sensitivity of A-T carriers. They may also show elevated clinical radiosensitivity. We retrospectively examined patients who presented with severe adverse reactions during or after standard radiation treatment for mutations in the gene responsible for A-T, ATM, considering a potential means of future identification of radiosensitive individuals prospectively to adjust dosage schedules. Material and Methods: We selected 20 cancer patients (breast, 11; rectum, 2; ENT, 2; bladder, 1; prostate, 1; anus, 1; astrocytoma, 1; Hodgkins lymphoma, 1) with Grade 3 to 4 (RTOG) acute and/or late tissue radiation side effects by reaction severity. DNA from the ...

1999-07-15

62

Density functional theory and topological analysis on the hydrogen bonding interactions in cysteine-thymine complexes  

British Library Electronic Table of Contents (United Kingdom)

Abstract Hydrogen bonding interactions between amino acids and nucleic acid bases constitute the most important interactions responsible for the specificity of protein binding. In this study, complexes formed by hydrogen bonding interactions between cysteine and thymine have been studied by density functional theory. The relevant geometries, energies, and IR characteristics of hydrogen bonds (H-bonds) have been systematically investigated. The quantum theory of atoms in molecule and natural bond orbital analysis have also been applied to understand the nature of the hydrogen bonding interactions in complexes. More than 10 kinds of H-bonds including intra- and intermolecular H-bonds have been found in complexes. Most of intermolecular H-bonds involve O (or N) atom as H-acceptor, whereas the...

2011-01-01

63

Chronic irradiation and brain development. Final (4th) progress report  

International Nuclear Information System (INIS)

The possible long term effects of chronic radiation exposure have been studied. Rats were given tritiated drinking water (3#mu#Ci/ml) before pregnancy, during pregnancy and threafter, continuously through 5 generations. The brains were studied at 30 and 120 days postnatally. Significant decreases in weight, DNA and protein contents of specific parts of the brain were found, the most pronounced decreases being in the diencephalon. The decreases in protein content were more pronounced than the decreases in DNA content; the decreases were also more pronounced at 30 days than at 120 days, and more pronounced in the fifth generation than in previous generations. An attempt was made to explain these complex phenomena on the basis of known periods of proliferation of various brain cells, damage to mothers themselves in each generation, and damage and repair of nucleic acids caused by radioactivity.

64

Biological radiation effects  

International Nuclear Information System (INIS)

The stages of processes leading to radiation damage are studied, as well as, the direct and indirect mechanics of its production. The radiation effects on nucleic acid and protein macro moleculas are treated. The physical and chemical factors that modify radiosensibility are analysed, in particular the oxygen effects, the sensibilization by analogues of nitrogen bases, post-effects, chemical protection and inherent cell factors. Consideration is given to restoration processes by excision of injured fragments, the bloching of the excision restoration processes, the restoration of lesions caused by ionizing radiations and to the restoration by genetic recombination. Referring to somatic effects of radiation, the early ones and the acute syndrome of radiation are discussed. The difference of radiosensibility observed in mammalian cells and main observable alterations in tissues and organs are commented. Referring to delayed radiation effects, carcinogeneses, ...

1976-01-01

65

Detecting and Genotyping Escherichia coli O157:H7 using multiplexed PCR and nucleic acid microarrays  

Energy Technology Data Exchange (ETDEWEB)

Rapid detection and characterization of food borne pathogens such as Escherichia coli O157:H7 is crucial for epidemiological investigations and food safety surveillance. As an alternative to conventional technologies, we examined the sensitivity and specificity of nucleic acid microarrays for detecting and genotyping E. coli O157:H7. The array was composed of oligonucleotide probes (25-30 mer) complementary to four virulence loci (intimin, Shiga-like toxins I and II, and hemolysin A). Target DNA was amplified from whole cells or from purified DNA via single or multiplexed polymerase chain reaction (PCR), and PCR products were hybridized to the array without further modification or purification. The array was 32-fold more sensitive than gel electrophoresis and capable of detecting amplification products from < 1 cell equivalent of genomic DNA (1 fg). Immunomagnetic capture, PCR and a microarray were subsequently used to detect 55 CFU ml-1 (E. coli O157:H7) ...

2000-12-01

66

A high stopping power, large acceptance Bragg curve spectrometer for heavy ion detection  

Energy Technology Data Exchange (ETDEWEB)

A large acceptance, high stopping power Bragg curve spectrometer has been developed for use in coincidence experiments with heavy ions. The electron collection fields are radial and position information is obtained from a resistive anode. The detector is 60 cm deep and operates at pressures of up to 2.5 atm of P-10 gas. It is mated to a scattering chamber which allows it to be moved out of plane during coincidence measurements. Factors affecting the performance of the detector are discussed, and results obtained with the detector are presented. (orig.).

1991-05-01

67

A high stopping power, large acceptance Bragg curve spectrometer for heavy ion detection  

International Nuclear Information System (INIS)

A large acceptance, high stopping power Bragg curve spectrometer has been developed for use in coincidence experiments with heavy ions. The electron collection fields are radial and position information is obtained from a resistive anode. The detector is 60 cm deep and operates at pressures of up to 2.5 atm of P-10 gas. It is mated to a scattering chamber which allows it to be moved out of plane during coincidence measurements. Factors affecting the performance of the detector are discussed, and results obtained with the detector are presented. (orig.).

68

Containment of genetically engineered organisms after application to subsurface environments. Technical completion report  

Energy Technology Data Exchange (ETDEWEB)

The feasibility of containing genetically engineered bacteria with enhanced dehalogenating properties for in situ bioremediation was investigated. (1) An agarose matrix microbead protocol and a detection system for contained microorganisms or DNA were developed. Multiplex Polymerase Chain Reaction (PCR) allowed tracking of a consortium of encapsulated organisms or several gene targets from a single species. Gene sequences encoding the enzymes responsible for initiating the biodegradation of toluene, octane, and 2,4-D were detected by multiplex PCR and nucleic acid probes from similar to 1-10 biodegradative cells/g soil. Improved DNA extraction methods resulted in PCR reactions detecting similar to 6 cells/g soil. (2) The pcpB gene (for the broad-spectrum detoxicant pentachlorophenol (PCP) hydroxylase) isolated from Flavobacterium sp. strain ATCC 39723 was used in attempts to develop an improved dehalogenating recombinant microorganism for containment experiments.

1993-06-01

69

Biophysical analysis of DNA modified by 1,2-diaminocyclohexane platinum(II) complexes  

Energy Technology Data Exchange (ETDEWEB)

Modification of DNA and double-stranded deoxyoligonucleotides with antitumor 1,2-diamino-cyclohexanedinitroplatinum(II) (Pt-dach) complexes was investigated with the aid of physico-chemical methods and chemical probes of nucleic acid conformation. The three Pt-dach complexes were used which differed in isomeric forms of the dach non-leaving ligand-Pt(1R,2R-dach), Pt(1S,2S-dach) and Pt(1R,2S-dach) complexes. The latter complex has lower antitumor activity than the other two Pt-dach complexes. Pt(1R,2S-dach) complex exhibits the slowest kinetics of its binding to DNA and of the conversion of monofunctional binding to bifunctional lesions. The anomalously slow electrophoretic mobility of multimers of the platinated and ligated oligomers suggests that bifunctional binding of Pt-dach complexes to a d(GG) site within double-stranded oligonucleotides induces bending of the oligomer. It is proposed that different effects of the Pt-dach complexes on DNA observed in this ...

1992-01-25

70

A DC-81-indole conjugate agent suppresses melanoma A375 cell migration partially via interrupting VEGF production and stromal cell-derived factor-1a-mediated signaling  

British Library Electronic Table of Contents (United Kingdom)

Pyrrolo[2,1-c][1,4]benzodiazepine (PBD) chemicals are antitumor antibiotics inhibiting nucleic acid synthesis. An indole carboxylate-PBD hybrid with six-carbon spacer structure (IN6CPBD) has been previously demonstrated to induce melanoma cell apoptosis and reduce metastasis in mouse lungs. This study aimed at investigating the efficacy of the other hybrid compound with four-carbon spacer (IN4CPBD) and elucidating its anti-metastatic mechanism. Human melanoma A375 cells with IN4CPBD treatment underwent cytotoxicity and apoptosis-associated assays. Transwell migration assay, Western blotting, and ELISA were used for mechanistic study. IN4CPBD exhibited potent melanoma cytotoxicity through interrupting G1/S cell cycle progression, increasing DNA fragmentation and hypodipoidic DNA contents, a...

2011-01-01

71

The Formation of the Low-sized High Density Plasma Structures in the Self-maintained Plasma-beam Discharge  

International Nuclear Information System (INIS)

The opportunity of use self-maintained plasma-beam discharge in an extended pulsing plasma diode of low pressure for making powerful sources of the soft X-rays is investigated. Conditions of formation of the self-maintained plasma-beam discharge are determined. The mode of making of dense high-temperature plasma on the basis of stannum ions in the discharge is shown. The stannum ions are used as a working element of a radiation sources at pulsing power of electron beam P?10...100 MW. Results of the examination on formation of the dense (np?1016 cm3), small sizes (le?100 eV in conditions of working material evaporation from the anode are given. The total contribution of energy to the discharge has made W < 20 J.

2006-01-01

72

Status report about the revision of an IEC standard for personal dose equivalent meters for H{sub p}(10) and H{sub p}(0,07); Statusbericht ueber die Neufassung einer IEC-Norm fuer H{sub p}(10)- und H{sub p}(0,07)-Personendosimeter  

Energy Technology Data Exchange (ETDEWEB)

A working group (SC 45B/WG B8) exists within the international electrochemical commission (IEC) which is working on the combination and revision of the IEC standards 61283, 61323, 61344, 61525 and 61526. All these standards concern direct reading, electronic, personal dose equivalent meters for the quantities H{sub p}(10) and H{sub p}(0,07); they are different with regard to the kind of radiation (photons, and/or beta rays and/or neutrons). The new standard with the number 61526 shall cover the following topics: 1. Combination of all the mentioned standards; 2. Maintenance of the essential requirements of the former standards; 3. Inclusion of the determination of the uncertainty of measurement for the measured dose equivalent value according to the ''Guide to the Expression of Uncertainty in Measurement''; 4. Consideration of all the relevant ISO-standards for reference radiation fields for photon, beta and ...

2001-07-01

73

Bragg Curve Counter for primary beam monitor  

Energy Technology Data Exchange (ETDEWEB)

The prototype Bragg Curve Counter (BCC) has been tested by {alpha}-source ({sup 241}Am) with a continuous gas flow system (P-10: 90% Ar+10% CH{sub 4}, at 300Torr). Two types of the Frisch grid were prepared for the BCC. One was made of expanded metal meshes, which consist of 175 meshes/inch{sup 2} with a 100{mu}m{sup t} nickel sheet. The other was made of a 1mm pitch wire with {phi} 50 {mu}m of the tungsten. The shape of Bragg peak signal was compared each other. It was found that the energy spectrum obtained by the wire grid is a little sharper than that obtained by the meshes grid. However the meshes grid is superior than the wire grid so far as handling and durability are concerned. (author).

1994-12-31

74

A large solid angle, high stopping power Bragg curve spectrometer for coincidence measurements  

Energy Technology Data Exchange (ETDEWEB)

A large acceptance, high stopping power, Bragg curve spectrometer has been developed for use in coincidence experiments with heavy ions. The electron collection fields are radial and position information is obtained from a resistive anode. The detector is 60 cm deep and operates at pressures of up to 2.5 atm of P-10 gas. It is mated to a scattering chamber which allows it to be moved out of plane during coincidence measurements. The detector design was aided by the results of computer simulations of the electron collection process in the detector, and of the signal processing in the electronics. The signals from the Bragg curve spectrometer are recorded in a waveform digitizer and the Bragg peak height, range, position and pileup rejection information are determined from software analysis of the recorded signals. Factors limiting the performance of the detector are discussed, and results obtained with the detector are presented. (orig.).

1991-09-01

75

A large solid angle, high stopping power Bragg curve spectrometer for coincidence measurements  

International Nuclear Information System (INIS)

A large acceptance, high stopping power, Bragg curve spectrometer has been developed for use in coincidence experiments with heavy ions. The electron collection fields are radial and position information is obtained from a resistive anode. The detector is 60 cm deep and operates at pressures of up to 2.5 atm of P-10 gas. It is mated to a scattering chamber which allows it to be moved out of plane during coincidence measurements. The detector design was aided by the results of computer simulations of the electron collection process in the detector, and of the signal processing in the electronics. The signals from the Bragg curve spectrometer are recorded in a waveform digitizer and the Bragg peak height, range, position and pileup rejection information are determined from software analysis of the recorded signals. Factors limiting the performance of the detector are discussed, and results obtained with the detector are presented. (orig.).

76

The thermal neutron sensitivities of SEI and Toshiba FD-P10-7 radiophotoluminescent glass dosimeters  

Energy Technology Data Exchange (ETDEWEB)

The themal neutron sensitivities of SEI and Toshiba FD-P10-7 radiophotoluminescent silver activated metaphosphate glass dosimeters have been measured and studied theoretically. The graphite thermal neutron column attached to the AEEW reactor NESTOR was utilised as a source of thermal neutrons with minimal fast neutron and gamma-ray contamination. Both the neutron removal and gamma correction measurement techniques were applied and these gave consistent results. It was found that the thermal neutron sensitivity of the Toshiba dosimeters is comparable to the most neutron intensitive thermoluminescent materials available. Because of their larger size and higher silver concentration, however, SEI dosimeters are an order of magnitude more neutron sensitive. The neutron sensitivity of FD-7 glass has not been reported previously, whereas in the case of SEI glass, there exists a modest body of previous work on dosimeters of nominally identical ...

1990-01-01

77

The thermal neutron sensitivities of SEI and Toshiba FD-P10-7 radiophotoluminescent glass dosimeters  

International Nuclear Information System (INIS)

The themal neutron sensitivities of SEI and Toshiba FD-P10-7 radiophotoluminescent silver activated metaphosphate glass dosimeters have been measured and studied theoretically. The graphite thermal neutron column attached to the AEEW reactor NESTOR was utilised as a source of thermal neutrons with minimal fast neutron and gamma-ray contamination. Both the neutron removal and gamma correction measurement techniques were applied and these gave consistent results. It was found that the thermal neutron sensitivity of the Toshiba dosimeters is comparable to the most neutron intensitive thermoluminescent materials available. Because of their larger size and higher silver concentration, however, SEI dosimeters are an order of magnitude more neutron sensitive. The neutron sensitivity of FD-7 glass has not been reported previously, whereas in the case of SEI glass, there exists a modest body of previous work on dosimeters of nominally identical ...

78

[Development of efficient DNA isolation procedures for Cryptosporidium and Trichinella PCR detection in fecal samples].  

Science.gov (United States)

PCR detection of genetic material of the parasites present in faeces may be an alternative for microscopic and serological tests routinely used for diagnosing parasitic enteral infections. However, small amount of target DNA combined with low efficiency of total DNA extraction, and presence of PCR inhibitors in the samples to be amplified, may cause false negative detection results. The aim of this work was to evaluate the impact of DNA isolation procedure used on the amplification of DNA fragments from the genomes of protozoan Cryptosporidium parvum and the nematode Trichinella spiralis. Two methods based on different principles of biological material lysis were evaluated; NucliSENS miniMAG employing simultaneously applied chemical lysis and mechanical disruption or mechanical disruption followed by enzymatic lysis in case of QIAamp DNA Stool Mini Kit. Both of the analyzed systems for nucleic acids purification allowed isolation of DNA from purified ...

2009-01-01

79

Tests for the diagnosis of Clostridium difficile infection: the next generation.  

Science.gov (United States)

Clostridium difficile (C. difficile) causes 25-30% of cases of antibiotic associated diarrhea and most cases of pseudomembranous colitis. Patients presenting with diarrhea after hospitalization for 3 or more days should be tested for C. difficile. There are many options available for testing, each of which has inherent advantages and disadvantages. Most laboratories perform toxin testing using an enzyme immunoassay method. In general these tests have sensitivities ranging from 60 to 70% and specificities of 98%. When using these methods, symptomatic patients with negative tests should be tested by another more sensitive method. Until recently, cell culture cytotoxicity neutralization assays (CCNAs) were considered the gold standard in the U.S. A two-step algorithm using an EIA for glutamate dehydrogenase detection followed by testing positives using CCNA, offered an improved alternative until the availability of molecular assays. Although early studies that compared the GDH assay to ...

2011-03-03

80

Regulation of electricity prices?  

International Nuclear Information System (INIS)

In this paper author deals with the regulation of electricity prices in the Slovak Republic. Author contests the social policy of the government through doped prices of electricity. Two thirds of electricity is generated in nuclear power plants in Slovakia. Hence, it is necessary to focus on the solution of problem of nuclear waste. In 2004 Ministry of Economy stated, that the deficit in nuclear fund, from which the country have to fully cover the costs of liquidation and final disposal of nuclear waste, is estimated in the amount of around 89 billion Slovak crowns (? 3.7 billion $). From it, so called historical deficit, which originated because of late foundation of fund, represents officially 15 billion Slovak crowns (? 0.62 billion $). In Slovakia exists the real risk, that by maintenance of present state by creation and draw of the fund, it will be possible to ensure only 39 per cent of financial sources necessary for full financial handling of the back part of nuclear energetic. ...

2006-12-21

 
 
 
 
81

One-step purification of Taq DNA polymerase using nucleotide-mimetic affinity chromatography.  

Science.gov (United States)

The thermostable Thermus aquaticus DNA polymerase (Taq Pol) has been the key factor in transforming the initial PCR method into one with huge impact in molecular biology and biotechnology. Therefore, the development of effective affinity adsorbents for the purification of Taq Pol, as well as other DNA polymerases, attracts the attention of the enzyme manufacturers and the research laboratories. In this report we describe a simple protocol for the purification of Taq Pol from E. coli lysates, leading to enzymes of high specific activity and purity. The protocol is based on a single affinity chromatography step, featuring an immobilized ligand selected from a structure-biased combinatorial library of dNTP-mimetic synthetic ligands. The ligand library was screened for its ability to bind and purify Taq Pol from E. coli lysates. One immobilized ligand (mABSGu) of the general formula X-Trz-Y, bearing 9-aminoethylguanine (AEGu) and aniline-2-sulfonic acid (mABS) linked on the triazine ...

2007-01-01

82

GLUE-IT and PEDEL-AA: new programmes for analyzing protein diversity in randomized libraries.  

Science.gov (United States)

There are many methods for introducing random mutations into nucleic acid sequences. Previously, we described a suite of programmes for estimating the completeness and diversity of randomized DNA libraries generated by a number of these protocols. Our programmes suggested some empirical guidelines for library design; however, no information was provided regarding library diversity at the protein (rather than DNA) level. We have now updated our web server, enabling analysis of translated libraries constructed by site-saturation mutagenesis and error-prone PCR (epPCR). We introduce GLUE-Including Translation (GLUE-IT), which finds the expected amino acid completeness of libraries in which up to six codons have been independently varied (according to any user-specified randomization scheme). We provide two tools for assisting with experimental design: CodonCalculator, for assessing amino acids corresponding to given randomized codons; and AA-Calculator, for finding ...

2008-04-28

83

Fossil avian eggshell preserves ancient DNA  

DEFF Research Database (Denmark)

Owing to exceptional biomolecule preservation, fossil avian eggshell has been used extensively in geochronology and palaeodietary studies. Here, we show, to our knowledge, for the first time that fossil eggshell is a previously unrecognized source of ancient DNA (aDNA). We describe the successful isolation and amplification of DNA from fossil eggshell up to 19 ka old. aDNA was successfully characterized from eggshell obtained from New Zealand (extinct moa and ducks), Madagascar (extinct elephant birds) and Australia (emu and owl). Our data demonstrate excellent preservation of the nucleic acids, evidenced by retrieval of both mitochondrial and nuclear DNA from many of the samples. Using confocal microscopy and quantitative PCR, this study critically evaluates approaches to maximize DNA recovery from powdered eggshell. Our quantitative PCR experiments also demonstrate that moa eggshell has approximately 125 times lower bacterial load than bone, making it a highly ...

2010-01-01

84

Effects of chronic ingestion of tritiated water on prenatal brain development  

International Nuclear Information System (INIS)

In view of the anticipated increased use of atomic energy in industry, the possible long-term effects of chronic radiation exposure were studied in five generations of rats. Female rats (F_0) were given tritiated drinking water ("3HOH;3#mu#Ci/ml) from adolescence (60 days) until and throughout pregnancy. A separate study showed that the maximum radioactivity in the urine is reached in 30 days, and in the blood in 42 days. In the newborns, the highest specific activity was in the nucleic acid fraction, but total radioactivity was mainly due to the water (body fluids) compartment. No signs of radiation illness or increase in cataract formation in the mothers were observed. The food and water intake and body weight changes before pregnancy were normal. The course and the outcome of pregnancy were also normal. However, 60% of the newborns (F_1) exhibited hematomas, edemas, and subdural hemorrhages, which disappeared at 30 days of age. Bollod analysis of the F_1 ...

85

Dose and temporary characteristics of the changes in the amount of nucleic acids in leucocytes of irradiated rats  

International Nuclear Information System (INIS)

An attempt was made for early quantitative assessment of the severety of an injury, using the nuclear acids (NA) content as a biochemical indicator. Sexually mature male rats with body mass 160-180 g were irradiated with doses 0.5, 1, 2, 4 and 8 Gy of "1"3"7Cs source. Uneven irradiation was achieved by shielding the abdominal-lumbar area with lead rings, measuring 2,5/50 mm. A 28-30% fall of the dose beneath the shielded zone was thus provided. Dose-dependent quantitative changes in Na content were determined on hours 24 and 72 within the range from 0.5 to 8 Gy and followed up until day 30 after irradiation with nonlethal (2 Gy) and lethal (8 Gy) doses. The total NA content in the leucocytes was determined by the method of Kritski and Aleksandrov. The uneven irradiation of rats within the 0,5-8 Gy range induced early dose-dependent changes in NA content. The nature of their post-irradiation dynamics also depended on the radiation dose and adequately reflected the periods of development ...

86

Dating divergences in the Fungal Tree of Life: review and new analyses.  

Science.gov (United States)

The collection of papers in this issue of Mycologia documents considerable improvements in taxon sampling and phylogenetic resolution regarding the Fungal Tree of Life. The new data will stimulate new attempts to date divergences and correlate events in fungal evolution with those of other organisms. Here, we review the history of dating fungal divergences by nucleic acid variation and then use a dataset of 50 genes for 25 selected fungi, plants and animals to investigate divergence times in kingdom Fungi. In particular, we test the choice of fossil calibration points on dating divergences in fungi. At the scale of our analysis, substitution rates varied without showing significant within-lineage correlation, so we used the Langley-Fitch method in the R8S package of computer programs to estimate node ages. Different calibration points had a dramatic effect on estimated divergence dates. The estimate for the age of the Ascomycota/Basidiomycota split was 1808000000 y ...

87

Dynamic {sup 31}P-MR-spectroscopy of the quadriceps muscle. Influence of sex and age on spectroscopic results; Die dynamische 31-Phosphor-Magnetresonanz-Spektroskopie des M. quadriceps. Einfluss von Geschlecht und Alter auf spektroskopische Parameter  

Energy Technology Data Exchange (ETDEWEB)

Purpose: {sup 31}P-MRS is used to assess the influence of sex and age on quadriceps muscle metabolism before and after exercise. Materials and Methods: 32 healthy volunteers (15 women, 17 men; mean age: 38{+-}17 yrs.) were examined by dynamic phosphorus-31 ({sup 31}P) magnetic resonance spectroscopy (MRS). In the magnet, the quadriceps muscle was stressed by an isometric and an isotonic form of exercise until exhaustion, respectively. Results: Resting conditions: With increasing subjects` age, the ratio {beta}-adenosine triphosphate/total phosphate decreased (r=-0.37; p=0.02). With increasing subjects` age, the ratios inorganic phosphate/phosphocreatine (r=0.79; p=5x10{sup -8}), phosphomonoester/{beta}-adenosine triphosphate (r=0.74; p=10{sup -6}) and phosphodiester/{beta}-adenosine triphosphate (r=0.62; p=10{sup -4}) increased. The pH was the only one of the evaluated spectroscopic parameters which ...

1999-05-01

88

High mobility two-dimensional hole system in GaAs/AlGaAs quantum wells grown on (100) GaAs substrates  

International Nuclear Information System (INIS)

We report on the transport properties of a high mobility two-dimensional hole system (2DHS) confined in GaAs/AlGaAs quantum wells grown by molecular-beam epitaxy on the (100) surface of GaAs. The quantum wells are modulation doped with carbon utilizing a resistive filament source. At T=0.3 K and carrier density p=1x10"1"1 cm"-"2, a mobility of 10"6 cm"2/Vs is achieved. At fixed carrier density p=10"1"1 cm"-"2, the mobility is found to be a nonmonotonic function of the quantum well width. The mobility peaks at 10"6 cm"2/Vs for a 15-nm well and is reduced for both smaller and larger well widths for these (100) samples. The mobility anisotropy is found to be small. Mobility along [011] is approximately 20% higher than along the [011] direction. In addition, the low-temperature carrier density is found to have low sensitivity to light. The hole density increases by only #approx#10% after exposure to red light at T=4.2 K. In structures designed for ...

2005-04-18

89

DYNAMICS OF SOLIDS IN THE MIDPLANE OF PROTOPLANETARY DISKS: IMPLICATIONS FOR PLANETESIMAL FORMATION  

International Nuclear Information System (INIS)

We present local two-dimensional and three-dimensional hybrid numerical simulations of particles and gas in the midplane of protoplanetary disks (PPDs) using the Athena code. The particles are coupled to gas aerodynamically, with particle-to-gas feedback included. Magnetorotational turbulence is ignored as an approximation for the dead zone of PPDs, and we ignore particle self-gravity to study the precursor of planetesimal formation. Our simulations include a wide size distribution of particles, ranging from strongly coupled particles with dimensionless stopping time #tau#_s #ident to# #OMEGA#t_s_t_o_p = 10"-"4 (where #OMEGA# is the orbital frequency, t_s_t_o_p is the particle friction time) to marginally coupled ones with #tau#_s = 1, and a wide range of solid abundances. Our main results are as follows. (1) Particles with #tau#_s #approx#> 10"-"2 actively participate in the streaming instability (SI), generate turbulence, and maintain the ...

2010-10-20

90

A genetic model of melanoma tumorigenesis based on allelic losses  

Energy Technology Data Exchange (ETDEWEB)

Previous karyotypic studies have indicated a possible series of non-random chromosomal events involved in the progression of melanoma. We sought to define a model of melanocyte tumorigenesis by studying allelic deletions of polymorphic simple tandem repeat markers mapping to chromosome 1, 6q, 7, 9p, 10, 11, 17, and 21 in thirty matched pairs of melanoma and constitutional DNAs. The most frequent and earliest deletions were found on 9p (57%) and 10q (32%) and with the exception of one case, no sample has loss of markers on another chromosome without concomitant loss of markers on 9p and/or 10q. Losses on 6q were also a frequent (32%) event that sometimes occurred in primary melanomas, whereas losses of loci on distal 1p (26%) or 11q (26%) occurred only in metastic melanomas. A background rate (0-17%) of allele loss was seen on chromosomes 7, 17, and 21. Homozygous deletions in a panel of 31 melanoma cell lines were only detected for markers on ...

1994-09-01

91

Simulation of sludge deposit onto a 900 MW steam generator tubesheet with the 3D code GENEPI  

Energy Technology Data Exchange (ETDEWEB)

Heat transfer processes use fluids which are generally not pure and can react with transfer surfaces. These surfaces are subject to deposits which can be sediments harmful to heat transfer and to integrity of materials. For nuclear plant steam generators, sludge build-up accelerates secondary side corrosion by concentrating chemical species. A major safety problem involved with such a corrosion is the growing of circumferential cracks which are very difficult to detect and size with eddy current probes. With a view to understand and control this problem, it is necessary to develop a mathematical model for the prediction of sludge behavior in PWR steam generators. Based on fundamental principles, this work intends to use different models available in literature for the prediction of the phenomenon leading to the accumulation of sludge particles at the bottom (the tubesheet) of a PWR. For that, a three-dimensional simulation of magnetite particulate fouling with the finite elements code ...

1998-07-01