Oligonucleotide DNAprobes complementary to the hypervariable region of the 16S rRNA of Bacteroides forsythus were tested for their specificity and sensitivity against reference and clinical isolates...Full Text Available
The relative sensitivities of different protocols for detecting cytomegalovirus nucleic acid sequences in histological specimens, using a biotinylated cDNAprobe, were assessed. Several commonly used...Full Text Available
chipD is a web server that facilitates design of DNA oligonucleotide probes for high-density tiling arrays, which can be used in a number of genomic applications such as ChIP-chip or gene-expression...Full Text Available
We report a simple method for the label-free detection of double-stranded DNA using surface-enhanced Raman scattering (SERS). We prepared cetyltrimethylammonium bromide (CTAB)-capped silver nanoparticles and a DNA-nanoparticle complex by adding silver nanoparticles to {lambda}-DNA solutions. In the present study, the utilization of CTAB-capped silver nanoparticles facilitates the electrostatic interaction between DNA molecules and silver nanoparticles; at the same time, the introduction of DNA avoids adding aggregating agent for the formation of nanoparticle aggregates to obtain large enhancement of DNA, because the DNA acts as both the probe molecules and aggregating agent of Ag nanoparticles. Atomic force microscopy (AFM) studies show that the morphology of DNA-Ag nanoparticle complexes seems to be ...
DNA barcodes linked to genetic features greatly facilitate screening these features in pooled formats using microarray hybridization, and new tools are needed to design large sets of barcodes to allow...Full Text Available
Hairpin oligonucleotides were synthesized with stems ending in a double-stranded structure, which can be ligated to double-strand breaks in DNA, and with loops that contain nucleotides modified by the...Full Text Available
AbstractWe describe the fusion of enhanced green fluorescent protein to the C-terminus of the HsdS DNA sequence-specificity subunit of the Type I DNA modification methyltransferase M.EcoKI....Full Text Available
Two overlapping cDNA clones encoding human DNA topoisomerase II were identified by two independent methods. In one, a human cDNA library in phage {lambda} was screened by hybridization with a mixed oligonucleotide probe encoding a stretch of seven amino acids found in yeast and Drosophila DNA topoisomerase II; in the other, a different human cDNA library in a {lambda}gt11 expression vector was screened for the expression of antigenic determinants that are recognized by rabbit antibodies specific to human DNA topoisomerase II. The entire coding sequences of the human DNA topoisomerase II gene were determined from these and several additional clones, identified through the use of the cloned human TOP2 gene sequences as probes. Hybridization between the cloned sequences and mRNA and genomic ...
Specific {sup 32}P-labeled DNAprobes were prepared and used to evaluate the distribution of tetracycline resistance determinants carried by gram-negative enteric bacteria isolated from pigs in 3 swine herds with different histories of antibiotic exposure. Plasmid DNA, ranging in size from 2.1 to 186 Kb, was observed in over 84% of 114 isolates studied. Two of 78 tetracycline resistant strains did not harbor plasmids. The DNAprobes were isolated from plasmids pSL18, pRT29/Tn10, pBR322 and pSL106, respectively, and they represented class A, B, C and D tetracycline resistance determinants. Hybridization conditions using 0.5X SSPE at 65{degrees}C minimize cross-hybridization between the different class of tetracycline resistance genes. Cross-hybridization between class A and class C determinants could be distinguished by simultaneous comparison of the intensity of their hybridization ...
DNA from 48 hr germinated rice embryos was cut with restriction endonuclease Bam H1 and cloned to the Bam H1 site on plasmid pBR 322. The clones containing recombinant DNA were selected by their sensitivity to tetracycline and resistance to ampicillin. Using /sup 32/P-labelled rice embryos tRNA as a probe two clones were identified to contain tRNA genes by colony hybridization.
An electrochemical detection method for analyzing sequence-specific DNA using gold nanoparticle DNAprobes and subsequent signal amplification step by silver enhancement is described. The assay relies on the electrostatic adsorption of target oligonucleotides onto the sensing surface of the glassy carbon electrode (GCE) and its hybridization to the gold nanoparticle-labeled oligonucleotides DNAprobe. After silver deposition onto gold nanoparticles, binding events between probe and target were monitored by the differential pulse voltammetry (DPV) signal of the large number of silver atoms anchored on the hybrids at the electrode surface. The signal intensity difference permits to distinguish between the match of two perfectly matched DNA strands and the near-perfect match where just one base pair was wrong. Coupled with this ...
Hepatitis B virus (HBV) DNA integration has been shown to occur frequently in human hepatocellular carcinomas. The authors have investigated whether common cellular DNA domains might be rearranged, possibly by HBV integration, in human primary liver tumors. Unique cellular DNA sequences adjacent to an HBV integration site were isolated from a patient with hepatitis B surface antigen-positive hepatocellular carcinoma. These probes detected rearrangement of this cellular region of chromosomal DNA in 3 of 50 additional primary liver tumors studied. Of these three tumor samples, two contained HBV DNA, without an apparent link between the viral DNA and the rearranged allele; HBV DNA sequences were not detected in the third tumor sample. By use of a panel of somatic cell hybrids, these unique cellular DNA ...
Hepatitis B virus (HBV) DNA integration has been shown to occur frequently in human hepatocellular carcinomas. The authors have investigated whether common cellular DNA domains might be rearranged, possibly by HBV integration, in human primary liver tumors. Unique cellular DNA sequences adjacent to an HBV integration site were isolated from a patient with hepatitis B surface antigen-positive hepatocellular carcinoma. These probes detected rearrangement of this cellular region of chromosomal DNA in 3 of 50 additional primary liver tumors studied. Of these three tumor samples, two contained HBV DNA, without an apparent link between the viral DNA and the rearranged allele; HBV DNA sequences were not detected in the third tumor sample. By use of a panel of somatic cell hybrids, these unique cellular DNA ...
High molecular weight DNA from pleroceroid larvae of the tapeworm Spirometra mansonoides was purified from isolated nuclei by conventional techniques. The DNA so isolated has a melting temperature (Tm) of 87 degrees C and a guanine plus cytosine (G/C) content of 44%. 5-Methyl cytosine could not be detected in plerocercoid DNA by HPLC analysis of DNA hydrolysates, by radiolabeling 5'-termini of MspI digests with polynucleotide kinase, or by comparing restriction patterns generated by MspI and HpaII. Renaturation kinetics demonstrated that the genome of S. mansonoides contains repetitive as well as single copy sequences and has a genome size estimated at approx. 1.6 X 10(9) bp. Hybridization was carried out between plerocercoid DNA and cDNAs for human beta-actin, alpha-tubulin and growth hormone (hGH). Rationale for this analysis was based on known homologies among actin and tubulin ...
BackgroundMicroorganisms display vast diversity, and each one has its own set of genes, cell components and metabolic reactions. To assess their huge unexploited metabolic potential...Full Text Available
This report describes our initial attempts at the molecular characterization of a maize controlling element. We have prepared a cDNAprobe and used it to detect changes at a locus where Ds elements are found. Evidence of their presence are indicated by changes in the restriction patterns, but there is as yet no information on the physical nature of the controlling elements nor on the kinds of rearrangements they cause.
A method to control charge transfer reaction in DNA using hybrid nanometer-sized TiO{sub 2} nanoparticles was developed. In this system extended charge separation reflects the sequence of DNA and was measured using metallic silver deposition or by photocurrent response. Light-induced extended charge separation in these systems was found to be dependent on the DNA-bridge length and sequence. The yield of photocatalytic deposition of silver was studied in systems having GG accepting sites imbedded in AT runs at varying distances from the TiO{sub 2} nanoparticle surface. Weak distance dependence of charge separation indicative of a hole hopping through mediating adenine (A) sites was found. The quantum yield of silver deposition in the system having a GG accepting site placed 8.5 {angstrom} from the nanoparticle surface was found to be {Phi} = 0.70 (70%) and {Phi} = 0.56 (56%) for (A){sub n} and (AT){sub n/2} bridge, ...
The relationship between X chromosome-linked adrenoleukodystrophy and the red/green color pigment gene cluster on Xq28 was investigated in a large kindred. The DNA in a hemizygous male showed altered restriction fragment sizes compatible with at least a deletion extending from the 5[prime] end of the color pigment genes. Segregation analysis using a DNAprobe within the color pigment gene cluster showed significant linkage with adrenoleukodystrophy (logarithm of odds score of 3.19 at [theta] = 0.0). These data demonstrate linkage, rather than association, between a unique molecular rearrangement in the color pigment gene cluster and adrenoleukodystrophy. The DNA changes in this region are thus likely to be helpful for determining the location and identity of the responsible gene. 33 refs., 4 figs.
It was the aim of this study to specifically detect the DNA sequences for the bphC gene, the meta-cleavage enzyme of the aerobic catabolic pathway for biphenyl and polychlorinated biphenyl degradation, in aquatic sediments without prior cultivation of microorganisms by using extraction of total DNA, PCR amplification of bphC sequences, and detection with specific gene probes. The direct DNA extraction protocol used was modified to enhance lysis efficiency. Crude extracts of DNA were further purified by gel filtration, which yielded DNA that could be used for the PCR. PCR primers were designed for conserved regions of the bphC gene from a sequence alignment of five known sequences. The specificity of PCR amplification was verified by using digoxigenin-labeled DNAprobes which were located internal to the amplified gene ...
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 ...
Two sequences with homology to a thioredoxin oligonucleotide probe were detected by Southern blot analysis of Anabaena sp. strain PCC 7120 genomic DNA. One of the sequences was shown to code for a protein...Full Text Available
Currently, over 30,000 mapped STSs and 27,000 mapped Unigenes (non-redundant, unigene sets of cDNA representing EST clusters) are available for human alone. A total of 44,000 Unigene cDNA clones have been supplied by Research Genetics. Unigenes, or cDNAs are excellent resource for map building for two reasons. Firstly, they exist in two alternative forms -- as both sequence information for PCR primer pairs, and cDNA clones -- thus making library screening by colony hybridization as well as pooled library PCR possible. The authors have developed an efficient and robust procedure to screen genomic libraries with large number of DNAprobes. Secondly, the linkage and order of expressed sequences, or genes are highly conserved among human, mouse and other mammalian species. Therefore, mapping with cDNA markers rather than random anonymous STSs will greatly facilitate ...
A {lambda}gt11 cDNA library was constructed from poly(U)-Spharose-selected Entamoeba histolytica trophozoite RNA in order to clone and identify surface antigens. The library was screened with rabbit polyclonal anti-E. histolytica serum. A 700-base-pair cDNA insert was isolated and the nucleotide sequence was determined. The deduced amino acid sequence of the cDNA revealed a cysteine-rich protein. DNA hybridizations showed that the gene was specific to E. histolytica since the cDNAprobe reacted with DNA from four axenic strains of E. histolytica but did not react with DNA from Entamoeba invadens, Acanthamoeba castellanii, or Trichomonas vaginalis. The insert was subcloned into the expression vector pGEX-1 and the protein was expressed as a fusion with the C terminus of glutathione S-transferase. Purified fusion protein ...
A #lambda#gt11 cDNA library was constructed from poly(U)-Spharose-selected Entamoeba histolytica trophozoite RNA in order to clone and identify surface antigens. The library was screened with rabbit polyclonal anti-E. histolytica serum. A 700-base-pair cDNA insert was isolated and the nucleotide sequence was determined. The deduced amino acid sequence of the cDNA revealed a cysteine-rich protein. DNA hybridizations showed that the gene was specific to E. histolytica since the cDNAprobe reacted with DNA from four axenic strains of E. histolytica but did not react with DNA from Entamoeba invadens, Acanthamoeba castellanii, or Trichomonas vaginalis. The insert was subcloned into the expression vector pGEX-1 and the protein was expressed as a fusion with the C terminus of glutathione S-transferase. Purified fusion protein ...
An /alpha//sub 2/-adrenergic receptor subtype has been cloned from a human kidney cDNA library using the gene for the human platelet /alpha//sub 2/-adrenergic receptor as a probe. The deduced amino acid sequence resembles the human platelet /alpha//sub 2/-adrenergic receptor and is consistent with the structure of other members of he family of guanine nucleotide-binding protein-coupled receptors. The cDNA was expressed in a mammalian cell line (COS-7), and the /alpha//sub 2/-adrenergic ligand (/sup 3/H)rauwolscine was bound. Competition curve analysis with a variety of adrenergic ligands suggests that this cDNA clone represents the /alpha//sub 2/B-adrenergic receptor. The gene for this receptor is on human chromosome 4, whereas the gene for the human platelet /alpha//sub 2/-adrenergic receptor (/alpha//sub 2/A) lies on chromosome 10. This ability to express the receptor in mammalian cells, free of other ...
Screening of a human placenta lambdagt11 library has led to the isolation of the cDNA for the human ..beta../sub 1/-adrenergic receptor (..beta../sub 1/AR). Used as the probe was the human genomic clone termed G-21. This clone, which contains an intronless gene for a putative receptor, was previously isolated by virtue of its cross hybridization with the human ..beta../sub 2/-adrenergic receptor (..beta../sub 2/AR). The 2.4-kilobase cDNA for the human ..beta../sub 1/AR encodes a protein of 477 amino acid residues that is 69% homologous with the avian ..beta..AR but only 54% homologous with the human ..beta../sub 2/AR. This suggests that the avian gene encoding ..beta..AR and the human gene encoding ..beta../sub 1/AR evolved from a common ancestral gene. RNA blot analysis indicates a message of 2.5 kilobases in rat tissues, with a pattern of tissue distribution consistent with ..beta../sub 1/AR binding. This pattern is quite ...
A cDNA library was constructed using RNA isolated from the livers of chickens which had been treated with zinc. This library was screened with a RNA probe complementary to mouse metallothionein-I (MT), and eight chicken MT cDNA clones were obtained. All of the cDNA clones contained nucleotide sequences homologous to regions of the longest (375 bp) cDNA clone. The latter contained an open reading frame of 189 bp, and the deduced amino acid sequence indicates a protein of 63 amino acids of which 20 are cysteine residues. Amino acid composition and partial amino acid sequence analyses of purified chicken MT protein agreed with the amino acid composition and sequence deduced from the cloned cDNA. Amino acid sequence comparison establish that chicken MT shares extensive homology with mammalian MTs. Southern blot analysis of chicken DNA indicates ...
The Drosophila sequence-specific DNA binding protein, Adf-1, is capable of activating transcription of the alcohol dehydrogenase gene, Adh, and is implicated in the transcriptional control of other developmentally regulated genes. We have cloned the cDNA encoding Adf-1 by generating specific DNAprobes deduced from partial amino acid sequence of the protein. Several cDNA clones encoding an extended open reading frame were isolated from a phage lambda library. The complete amino acid sequence of Adf-1 deduced from the longest cDNA reveals structural similarities to the putative helix-turn-helix DNA binding motif of Myb and Myb-related proteins. DNA sequence analysis of genomic clones and Northern blot analysis of mRNA suggest that Adf-1 is a single-copy gene encoding a 1.9-kb transcript. Purified recombinant Adf-1 ...
The DNA sequence motif ATTTGCAT (octamer) or its inverse complement has been identified as an evolutionarily conserved element in the promoter region of immunoglobulin genes. Two major DNA-binding proteins that bind in a sequence-specific manner to the octamer DNA sequence have been identified in mammalian species--a ubiquitously expressed protein (Oct-1) and a lymphoid-specific protein (Oct-2). During characterization of the promoter region of the chicken immunoglobulin light chain gene, the authors identified two homologous octamer-binding proteins in chicken B cells. when the cloning of the human gene for Oct-2 revealed it to be a member of a distinct family of homeobox genes, they sought to determine if the human Oct-2 cDNA could be used to identify homologous chicken homeobox genes. Using a human Oct-2 homeobox-specific DNAprobe, they were able to identify ...
Introduction: Academic research has made a significant advancement in understanding the viral causes of cervical cancer and generating the technology for prevention, both at the primary and secondary levels. Human papillomaviruses (HPVs) have been recognized as the first necessary cause of cervical cancer, the second most common cancer in women worldwide. Areas covered: This paper reviews the epidemiological evidence of the causality of HPV in relation to cervical cancer, other genital tract cancers and some cancers of the oral cavity and oropharynx. The review also covers HPV DNA testing as a screening tool. DNAprobes of high-risk HPV types in different formats have been fully validated as primary screening tests, as secondary triage tests and as a prognostic marker following treatment o...
To more accurately measure fluorescent signals from microarrays, we calibrated our acquisition and analysis systems by using groundtruth samples comprised of known quantities of red and green gene-specific DNAprobes hybridized to cDNA targets. We imaged the slides with a full-field, white light CCD imager and analyzed them with our custom analysis software. Here we compare, for multiple genes, results obtained with and without preprocessing (alignment, color crosstalk compensation, dark field subtraction, and integration time). We also evaluate the accuracy of various image processing and analysis techniques (background subtraction, segmentation, quantitation and normalization). This methodology calibrates and validates our system for accurate quantitative measurement of microarrays. Specifically, we show that preprocessing the images produces results significantly closer to the known ground-truth for these samples.
We have isolated a genomic clone containing 'Lupinus luteus' 5S ribosomal RNA genes by screening with 5S rDNAprobe clones that were hybridized previously with the initiator methionine tRNA preparation (contaminated) with traces of rRNA or its degradation products). The clone isolated contains ten repeat units of 342 bp with 119 bp fragment showing 100% homology to the 5S rRNA from yellow lupine. Sequence analysis indicates only point heterogeneities among the flanking regions of the genes. (author). 6 refs, 3 figs.
pD10 is a 0.85 kb fragment with BamHI ends cloned in pSP64. It was sub-cloned from {lambda}D10 which was isolated from a genomic library constructed in {lambda}EMBL3 from a rodent human somatic cell hybrid. The probe was localized to 19q13.2-19qter on a panel of rodent human somatic cell hybrids. Co-dominant segregation was shown in 32 families of 280 individuals.
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 ...
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 ...
We aim to isolate cDNAs mapping to human chromosome 16 and localise such cDNAs on the high resolution physical map. In collaboration with LANL, PCR primers will be synthesised from cDNA sequences mapped to chromosome 16 and used as ESTs in the generation of mega-YAC contigs for this chromosome. Probing of high density cosmid grids will enable integration of the ESTs into cosmid contigs and location of the cosmid contigs on the YAC contig. A hn-cDNA library has been constructed from the hybrid CY18 which contains chromosome 16 as the only human chromosome. A modified screening protocol has been successfully developed and 15 hn-cDNA clones have been sequenced and localised on the hybrid map. Sequence analysis of four of these revealed that they were known cDNAs, which are now mapped to chromosome 16. Development of techniques to allow the isolation of longer cDNAs from the identified exons is in progress. ...
The antitumor agent camptothecin targets DNA topoisomerase I by reversibly stabilizing a covalent enzyme-DNA intermediate. The subsequent collision of DNA replication forks with these drug-enzyme-DNA...Full Text Available
We describe the cloning and characterization of a mutated thermostable DNA polymerase from Thermus aquaticus (Taq) that exhibits an increased reverse transcriptase activity and is therefore designated for one-step PCR pathogen detection using established real-time detection methods. We demonstrate that this Taq polymerase mutant (Taq M1) has similar PCR sensitivity and nuclease activity as the respective Taq wild-type DNA polymerase. In addition, and in marked contrast to the wild-type, Taq M1 exhibits a significantly increased reverse transcriptase activity especially at high temperatures (>60 degrees C). RNA generally hosts highly stable secondary structure motifs, such as hairpins and G-quadruplexes, which complicate, or in the worst case obviate, reverse transcription (RT). Thus, RT at high temperatures is desired to weaken or melt secondary structure motifs. To demonstrate the ability of Taq M1 for RNA detection of pathogens, we ...
The human smooth muscle myosin heavy chain locus (MYH11) was mapped by fluorescence in situ hybridization to the middle of the p arm of chromosome 16 using a genomic cosmid clone containing coding sequences of the gene as probe. Probe from coding sequence, when applied to Southern blots of a panel of hybrids containing different portions of human chromosome 16, localized the gene to 16p13.13-13.12. Coding sequence PCR primers, when used on the DNA from a CHO-mouse hybrid clone mapping panel informative for mouse chromosomes, showed that the gene was located on mouse chromosome 16. These results correct a recent assignment of MYH11 from 16q12.2 to the region of the 16p-arm inversion breakpoint seen in acute myelomonocytic leukemia (AMML) M4Eo and demonstrate that the conflicting data do not result from the presence of additional MYH genes on the q arm of the chromosome. Also, a new region of conserved synteny between human ...
The product of the human GRO gene is a cytokine with inflammatory and growth-regulatory properties; GRO is also called MGSA for melanoma growth-stimulatory activity. The authors have identified two additional genes, GRO#beta# and GRO#gamma#, that share 90% and 86% identity at the deduced amino acid level with the original GRO#alpha# isolate. One amino acid substitution of proline in GRO#alpha# by leucine in GRO#beta# and GRO#gamma# leads to a large predicted change in protein conformation. Significant differences also exist in the 3' untranslated region, including different numbers of ATTTA repeats associated with mRNA instability. A 122-base-pair region in the 3' region is conserved among the three GRO genes, and a part of it is also conserved in the Chinese hamster genome, suggesting a role in regulation. DNA hybridization with oligonucleotide probes and partial sequence analysis of the genomic clones confirm that the three forms are derived ...
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 ...
The authors used a near full-length human ornithine [delta]-aminotransferase cDNA, huOAT6, as a probe under low stringency hybridization conditions to identify a new autosomal ornithine [delta]-aminotransferase-related sequence (OATL3). Cloning and characterization of this sequence reveal it to be a partial nonprocessed pseudogene corresponding to exon 3 and flanking intronic sequences of the ornithine [delta]-aminotransferase structural gene. Using somatic cell hybrids and fluorescence in situ hybridization, they mapped OATL3 to 10q26, adjacent to the ornithine [delta]-aminotransferase structural gene locus. 13 refs., 3 figs.
The heat-shock gene, Hsp90, was targeted as a new variable genomic region to supplement other DNA-based tests for identification and discrimination of Globodera pallida, G. rostochiensis and G. tabacum tabacum. Populations of the potato cyst nematodes, G. pallida and G. rostochiensis (PCN), originating from Canada, France, Belgium and USA, together with two populations of G. tabacum tabacum from the USA and France were used for the amplification of a fragment of the Hsp90 gene. General and specific primers and probes for each species were derived from the consensus and non-consensus regions of the aligned sequences, respectively. A triplex conventional PCR assay, using a general forward and reverse or three specific reverse primers, as well as a real-time PCR using general primers and spec...
The astonishingly efficient location and excision of damaged DNA bases by DNA repair glycosylases is an especially intriguing problem in biology. One example is the enzyme uracil DNA glycosylase (UNG),...Full Text Available
Recently reported technologies for DNA-directed organic synthesis and for DNA computing rely on routing DNA populations through complex networks. The reduction of these ideas to practice has been limited...Full Text Available
A gene designated BRCA1, implicated in the susceptibility to early-onset familial breast cancer, has recently been localized to chromosome 17q12-q21. To date, the order of DNA markers mapped within this region has been based on genetic linkage analysis. The authors report the use of multicolor fluorescence in situ hybridization to establish a physically based map of five polymorphic DNA markers and 10 cloned genes spanning this region. Three cosmid clones and Alu-PCR-Generated products derived from 12 yeast artificial chromosome clones representing each of these markers were used in two-color mapping experiments to determine an initial proximity of markers relative to each other on metaphase chromosomes. Interphase mapping was then employed to determine the order and orientation of closely spaced loci by direct visualization of fluorescent signals following hybridization of three probes, each detected in a different color. ...
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 ...
Duchenne and Becker muscular dystrophies are caused by defects of dystrophin, which forms a part of the membrane cytoskeleton of specialized cells such as muscle. It has been previously shown that the dystrophin-associated protein A1 (59-kDa DAP) is actually a heterogeneous group of phosphorylated proteins consisting of an acidic ({alpha}-A1) and a distinct basic ({beta}-A1) component. Partial peptide sequence of the A1 complex purified from rabbit muscle permitted the design of oligonucleotide probes that were used to isolate a cDNA for one human isoform of A1. This cDNA encodes a basic A1 isoform that is distinct from the recently described syntrophins in Torpedo and mouse and is expressed in many tissues with at least five distinct mRNA species of 5.9, 4.8, 4.3, 3.1, and 1.5 kb. A comparison of the human cDNA sequence with the GenBank expressed sequence tag (EST) data base has identified a relative ...
The following topics are discussed: insertion of incorrect bases in DNA; ability of DNA polymerase to copy uv-irradiated DNA; role of enzymes in repair of DNA; effects of uv radiation on molecular weight of DNA; photoreactivation; repair of DNA in Escherichia coli and xp cells following uv radiation; and synchrotron radiation studies on DNA repair. (HLW)
Today you will learn about the parts of DNA and what DNA, genes and chromosomes are. Today you will learn what DNA, genes and chromosomes are and the parts of the DNA molecule. Look at all of the websites, take whatever notes you need to. At the end of the assignment, be able to describle DNA, the parts of DNA, genes and chromosomes. Covers Biology Core Curriculum, ...
Typing Workshop": [Introductions] [Intro, DNA Basics, and Historical Perspective] [DNA Extraction] [Validation and QA/QC] [DNA Quantitation] [PCR Amplification] [STR Loci and Kits]...
The main aim of DNA barcoding is to establish a shared community resource of DNA sequences that can be used for organismal identification and taxonomic clarification. This approach was successfully...Full Text Available
Abstract: Snake venom contains a number of small proteins,enzymes and other components,which displays a broad spectrum of biological activities. With the ability of specifically binding on acetylcholine acceptor, alpha-bungarotoxins are not only useful molecular probes in investigating the mechanism of neural signal transmission, but also potential pharmic preparations for neural disease treatment. In current research,cDNAs of Bungarus multicinutus venom gland were synthesized using SMART cDNA amplification kit and then, alpha-bungarotoxin genes were cloned and sequenced. Total of 20 clones were sequenced representing 14 isotoxin mRNAs of alpha-bungarotoxins. Among those clones, a novel isotoxin gene was subcloned into two expression plasmids, alpha-BgTX/pQE30a and alpha-BgTX/pGEX-4T-1, and transformed into E. coli. After inducing with IPTG, fused protein of GST-alpha-BgTX was successfully expressed at level of 30% gross proteins of bacteria. ...
To resolve the phylogeny of the autochthonous mitochondrial DNA (mtDNA) haplogroups of India and determine the relationship between the Indian and western Eurasian mtDNA pools more precisely, a diverse...Full Text Available
BackgroundDNA repair is the general term for the collection of critical mechanisms which repair many forms of DNA damage such as methylation or ionizing radiation. DNA repair has...Full Text Available
The presence of DNA-anti-DNA complexes in sera from patients with systemic lupus erythematosus (SLE) was investigated by two new radioimmunoassays (RIA) developed for this purpose and by measuring the...Full Text Available
The Tetrahymena thermophila origin recognition complex (ORC) contains an integral RNA subunit, 26T RNA, which confers specificity to the amplified ribosomal DNA (rDNA) origin by base...Full Text Available
Expression of IGF-2 and changes in its expression in response to systemic endocrine alterations have not been demonstrated for adipocytes. Adipocytes were induced to develop within cultures of 3T3-L1 cells using a medium containing 0.5mM isobutylmethylxanthine, 1uM insulin and 100ng hydrocortisone/ml for 48 hours of exposure. Cultures containing developing adipocytes were incubated with 10% pig serum and 1 uM insulin for several days. The resultant adipocyte cultures were then treated with either 10% pig serum, diabetic pig serum or hypophysectomized pig serum in DMEM for 48 hours. Adipocytes within the cultures were separated from undifferentiated cells using percoll density gradient centrifugation. Total RNA was isolated from adipocytes and dot blotted. Blots were probed with a {sup 32}P-cDNAprobe for rat IGF-2. IGF-2 was expressed by the adipocytes and the pattern of expression showed specific differences between serum ...
ADP-ribosylation factors (ARFs) are small guanine nucleotide-binding proteins that enhance the enzymatic activities of cholera toxin. Two ARF cDNAs, ARF1 and ARF3, were cloned from a human cerebellum library. Based on deduced amino acid sequences and patterns of hybridization of cDNA and oligonucleotide probes with mammalian brain poly(A)"+ RNA, human ARF1 is the homologue of bovine ARF1. Human ARF3, which differs from bovine ARF1 and bovine ARF2, appears to represent a newly identified third type of ARF. Hybridization patterns of human ARF cDNA and clone-specific oligonucleotides with poly(A)"+ RNA are consistent with the presence of at least two, and perhaps four, separate ARF messages in human brain. In vitro translation of ARF1, ARF2, and ARF3 produced proteins that behaved, by SDS/PAGE, similar to a purified soluble brain ARF. Deduced amino acid sequences of human ARF1 and ARF3 contain regions, similar to those in ...
Signal-to-noise-ratio (SNR) thresholds for microarray data analysis were experimentally determined with an oligonucleotide array that contained perfect match (PM) and mismatch (MM) probes based upon four genes from Shewanella oneidensis MR-1. A new SNR calculation, called signal to both standard deviations ratio (SSDR) was developed, and evaluated along with other two methods, signal to standard deviation ratio (SSR), and signal to background ratio (SBR). At a low stringency, the thresholds of SSR, SBR, and SSDR were 2.5, 1.60 and 0.80 with oligonucleotide and PCR amplicon as target templates, and 2.0, 1.60 and 0.70 with genomic DNA as target templates. Slightly higher thresholds were obtained at the high stringency condition. The thresholds of SSR and SSDR decreased with an increase in the complexity of targets (e.g., target types), and the presence of background DNA, and a decrease in the composition of targets, while SBR ...
New industrial Non Destructive Testing requires the development of new probes: technologies such as high spatial resolution probes for small breaking flaws detection or flexible probes for complex curved surfaces inspection. The CEA/LIST has designed and integrated 2 probes to answer industrial's constraints. The first one is based on Giant Magneto-Resistance and is able to detect 50 ?m3 flaws. The second one is based on micro-coils etched on kapton and its flexibility is adapted to curved surfaces inspection. Both are multi-elements probes, allowing fast inspection. This paper presents those new probes, their design and their performances.
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.
Nuclear steam generator tubes have become flawed in ways that challenge conventional eddy current probes. In response to the shortcomings of conventional probes, array probes have been developed to improve measurement capabilities. However, the commercially available array probes have exhibited several weaknesses that offset the advantages and limit the applications in steam generator inspections. A primary weakness is the relatively high rate of probe failure coupled with the high unit cost for each probe. This can be costly for a utility in the time lost for probe replacement and increased radiation exposure in addition to the probe costs. Other weaknesses which make array probes undesirable for routine use are: poor mechanical and electrical characteristics; difficulty in operation and ...
A multiple probe approach of implanting microdialysis probes into each separate tissue layer would better represent sampling from the stomach. Presently, microdialysis sampling experiments are...Full Text Available
The Lesch-Nyhan (LN) syndrome is a severe X chromosome-linked disease that results from a deficiency of the purine salvage enzyme hypoxanthine phosphoribosyltransferase (HPRT). The mutations leading to the disease are heterogeneous and frequently arise as de novo events. The authors have identified nucleotide alterations in 15 independently arising HPRT-deficiency cases by direct DNA sequencing of in vitro amplified HPRT cDNA. They also demonstrate that the direct DNA sequence analysis can be automated, further simplifying the detection of new mutations at this locus. The mutations include DNA base substitutions, small DNA deletions, a single DNA base insertion, and errors in RNA splicing. The application of these procedures allows DNA diagnosis and carrier identification by the direct detection of the mutant alleles within individual ...
The overall objective of this project is to produce antibodies to unique modified DNA bases and develop immunochemical assays to quantitate these lesions in damaged DNA. During this past year we have developed an antibody and chemical test to quantitate a basic sites in DNA and produced antibodies to the 8-oxopurines. This report discusses the detection of a basic sites in DNA and the preparation of antibodies to 8-hydroxyadenine and 8-hydroxyguanine.
Uracil appears in DNA as a result of cytosine deamination and by incorporation from the dUTP pool. As potentially mutagenic and deleterious for cell regulation, uracil must be removed from DNA....Full Text Available
Cytosine methylation in DNA is a major epigenetic signal, and plays a central role in propagating chromatin status during cell division. However the mechanistic links between DNA methylation...Full Text Available
We use X-ray scattering and molecular simulations to investigate the structural properties of complexes of multivalent cationic lipids and DNA molecules. At low mole fraction of neutral lipids (NLs), $\\Phi_{\\rm NL}$, the complexes show dramatic DNA compaction down to essentially close packed DNA arrays with a DNA interaxial spacing $d_{\\rm DNA}=25\\AA$. A gradual increase in $\\Phi_{\\rm NL}$ does not lead to a continuous increase in $d_{\\rm DNA}$ as observed for DNA complexes of monovalent cationic lipids (CLs). Instead, distinct spacing regimes exist, with sharp transitions between them. Three packing states have been identified: (i) close packed, (ii) condensed, but not close packed, with $d_{\\rm DNA}=27-28\\AA$, and (iii) an expanded state, where $d_{\\rm DNA}$ increases gradually with ...
Sequence-directed variations in the canonical DNA double helix structure that retain Watson-Crick base-pairing play important roles in DNA recognition, topology, and nucleosome positioning....Full Text Available
DNA double-strand breaks (DSBs) and other lesions occur frequently during cell growth and in meiosis. These are often repaired by homologous recombination (HR). HR may result in the formation of DNA...Full Text Available
This paper announces the availability of the first international standard for anti-double-stranded DNA (anti-dsDNA). The material, coded Wo/80, was obtained after recalcification of plasma taken from...Full Text Available
Hepatitis B virus (HBV) DNA integration has been shown to occur frequently in human hepatocellular carcinomas. We have investigated whether common cellular DNA domains might be rearranged, possibly...Full Text Available
The polymerase chain reaction (PCR) was investigated as a means of diagnosing Mycoplasma pneumoniae infections. The target DNA sequence was a 375-bp segment of the P1 virulence protein. This DNA segment...Full Text Available
The conformation of the self-complimentary DNA dodecamer d(br5CGbr5CGAATTbr5CGbr5CG) has been investigated in a variety of salt and solvent conditions by one and two-dimensional 1H NMR. In low salt...Full Text Available
The sequences of the internal transcribed spacer (ITS) ribosomal DNA (rDNA) domain data obtained by restriction fragment length polymorphism analysis with 18S rDNA and fingerprinting (M13) for clinical...Full Text Available
To gain insight into the process of mitochondrial transmission in yeast, we directly labeled mitochondrial proteins and mitochondrial DNA (mtDNA) and observed their fate after the fusion of two cells....Full Text Available
Mitochondrial DNA is constantly exposed to oxidative injury. Due to its location close to the main site of reactive oxygen species, the inner mitochondrial membrane, mtDNA is more susceptible...Full Text Available
BackgroundMitochondrial dysfunction, oxidative damage and the accumulation of somatic mutations in mitochondrial DNA (mtDNA) have been associated with certain neurodegenerative disorders....Full Text Available
Many genetic processes depend on proteins interacting with specific sequences on DNA. Despite the large excess of nonspecific DNA in the cell, proteins can locate their targets rapidly. After initial...Full Text Available
The effects of various fixative agents, pH, ionic strength, stain concentration, and magnesium concentration on DNA staining with the antibiotics olivomycin, chromomycin A3, and mithramycin were examined with DNA in solution and in mammalian cells.
Chromosomal translocation, deletion, and inversion/duplication directly linked to hepatitis B virus (HBV) DNA integration occur frequently in host DNA of human hepatocellular carcinomas. To test the...Full Text Available
SummaryDnaA initiates chromosomal replication in E. coli at a well regulated time in the cell-cycle. To determine how the spatial distribution of DnaA is related...Full Text Available
Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MS) has been explored widely for DNA sequencing. The major requirement for this method is that the DNA sequencing fragments...Full Text Available
CDC7 is an essential gene required for DNA replication in Saccharomyces cerevisiae. Cdc7p homologs have recently been identified in vertebrates, but their role in DNA...Full Text Available
DNA damage is a threat to genomic integrity in all living organisms. Plants and green algae are particularly susceptible to DNA damage especially that caused by UV light, due to their light dependency...Full Text Available
Competent Haemophilus cells recognize and preferentially take up Haemophilus DNA during genetic transformation. This preferential uptake is correlated with the presence on incoming DNA of an 11-base-pair...Full Text Available
It is well known that DNA strands bind to silica surfaces in the presence of high concentrations of chaotropic salts. We developed simple methods to evaluate binding and recovery of DNA on flat...Full Text Available
The standard paradigm postulates that the human mitochondrial genome (mtDNA) is strictly maternally inherited and that, consequently, mtDNA lineages are clonal. As a result of mtDNA clonality, phylogenetic...Full Text Available
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
A DNA fragment located on the 3' side of the Coxiella burnetii htpAB operon was determined by Southern blotting to exist in approximately 19 copies in the Nine Mile I genome. The DNA sequences of this...Full Text Available
The dependence of the mobility of DNA molecules through an hexagonal array of micropillars on their length and the applied electric field was investigated and it was found that mobility is a nonmonotonic function of their length. Results also revealed that the size dependence of the DNA mobility depends on the applied electric field and there is a crossover around E 25 V/cm for the mobility of l-DNA and T4-DNA. These observations are explained in terms of the diffusion process inside the structure affected by the solvent and are modeled using the Langevin and its corresponding Fokker-Planck equations. The phenomenon is generalized under three regimes in a phase diagram relating the electric field and the DNA lengths. The model and the associated phase diagram described here provide an expl...
This paper examines theoretically the effects that restraints on the tertiary structure of a superhelical DNA domain exert on the energetics of linking and the onset of conformational transitions. The...Full Text Available
Telomeres protect the chromosome ends from unscheduled DNA repair and degradation. Telomeres are heterochromatic domains composed of repetitive DNA (TTAGGG repeats) bound to an array of specialized...Full Text Available
These proceedings collect papers on the subject of lymphokines. Topics include: DNA-cloning of mouse and human lymphokine genes, inteferons, interleukins, gene expression, tumor necrosis factors, and recombinant DNA.
Metastable and somatically heritable patterns of DNA methylation provide an important level of genomic regulation. In this article, we review methods for analyzing these genome-wide epigenetic...Full Text Available
Since the elucidation of the structure of double helical DNA, the construction of small molecules that recognize and react at specific DNA sites has been an area of considerable interest. In...Full Text Available
The inflexibility of double-stranded DNA with respect to bending and twisting is well established in vitro. Understanding apparent DNA physical properties in vivo is...Full Text Available
BackgroundPublicly available DNA sequence databases such as GenBank are large, and are growing at an exponential rate. The sheer volume of data being dealt with presents serious...Full Text Available
BackgroundMitochondrial DNA (mtDNA) is being analyzed by an increasing number of laboratories in order to investigate its potential role as an active marker of tumorigenesis...Full Text Available
This paper describes various eddy current probes, and their performance, which were developed at the Chalk River Laboratories of Atomic Energy of Canada. Included are probes for detecting defects at tubesheet regions in heat exchanger tubes, defects at expansion-transition regions in finned tubes, and defects in ferromagnetic tubes. All of these probes can be used with conventional commercially available instruments. (author).
Described herein is an optical probe (120) for use in characterizing surface defects in wafers, such as semiconductor wafers. The optical probe (120) detects laser light reflected from the surface (124) of the wafer (106) within various ranges of angles. Characteristics of defects in the surface (124) of the wafer (106) are determined based on the amount of reflected laser light detected in each of the ranges of angles. Additionally, a wafer characterization system (100) is described that includes the described optical probe (120).
A new generation of transmit-receive single-pass probes, denoted as C6 or X probe, was field tested during the Gentilly II, 2000 steam generator tube inspection. This probe has a performance equivalent to rotating probes and can be used for tubesheet and full-length inspection at an inspection speed equivalent to that of bobbin probes. Existing C3 transmit-receive probes have been demonstrated to be effective in detecting circumferential cracks. The C5 probe can detect both circumferential and axial cracks and volumetric defects but cannot discriminate between them. The C6 probe expands on the capabilities of both probes in a single probe head. It can simultaneously detect and discriminate between circumferential and axial cracks to satisfy different plugging criteria. It has ...
The chain of eddy current probes and a film cassette probe, which responds to radiation from a radio-active source in one of the heat exchanger pipes is examined. The probes are moved by nylon pipes on both ends of the chain through the pipe to be examined. The nylon pipes are bent off load. In this way the probes on the chain will adjust near to the plane of bending of the pipes to be tested.
The helicase action of the Escherichia coli UvrAB complex on a covalently closed circular DNA template was monitored using bacterial DNA topoisomerase I, which specifically removes negative supercoils. In the presence of E. coli DNA topoisomerase I and ATP, the UvrAB complex gradually introduced positive supercoils into the input relaxed plasmid DNA template. Positive supercoils were not produced when E. coli DNA topoisomerase I was replaced by eukaryotic DNA topoisomerase I or when both E. coli and eukaryotic DNA topoisomerases I were added simultaneously. These results suggest that like other DNA helix-tracking processes, the ATP-dependent action of the UvrAM complex on duplex DNA simultaneously generates both positive and negative supercoils, which are not constrained by protein binding but are ...
Objective: To determine whether asymptomatic bacterial vaginosis (BV) is associated with upper genital tract inflammation or bacterial colonization.Methods: Fifty nonpregnant women with intact uteri who planned to undergo gynecologic surgery and had no recent symptoms of vaginal infection were enrolled. We obtained a vaginal swab for Gram stain, endocervical swab for chlamydia and gonorrhea DNAprobe testing, and Pipelle endometrial biopsy for aerobic and anaerobic cultures and histology. We correlated surgical findings and histology of available surgical specimens with the microbiologic results. The diagnosis of BV was made according to Speigel's criteria. Bacteria isolated from the uterus were classified as high virulence versus low virulence. Contingency tables were analyzed using the chi-square or Fisher Exact tests.Results: Twenty-one of 50 patients had BV on Gram stain, 3 had intermediate BV, and 3 had unreadable slides. Eleven patients ...
Nucleotide sequence analysis of a genomic clone from an Ashkenazic Jewish patient with type 1 Gaucher disease revealed a single-base mutation (adenosine to guanosine transition) in exon 9 of the glucocerebrosidase gene. This change results in the amino acid substitution of serine for asparagine. Transient expression studies following oligonucleotide-directed mutagenesis of the normal cDNA confirmed that the mutation results in loss of glucocerebrosidase activity. Allele-specific hybridization with oligonucleotide probes demonstrated that this mutation was found exclusively in type 1 phenotype. None of the 6 type 2 patients, 11 type 3 patients, or 12 normal controls had this allele. In contrast, 15 of 24 type 1 patients had one allele with this mutation, and 3 others were homozygous for the mutation. Furthermore, some of the Ashkenazic Jewish type 1 patients had only one allele with this mutation, suggesting that even in this population there is ...
The large aggregating proteoglycan aggrecan is a major structural component of the extracellular matrix of articular cartilage. Recent cDNA cloning of the human aggrecan gene (AGC1) reveals a core protein of at least 2316 amino acids characterized by several distinct structural domains. Two globular domains, termed G1 and G2, are present at the amino terminus of the molecule and a third, termed G3, is present at the carboxy terminus. The G1 domain is homologous in structure to the cartilage link protein and accounts for the aggregating potential of aggrecan through its ability to interact with hyaluronic acid. The aggrecan gene is known to consist of 15 exons, with each exon encoding a distinct functional region of the mature protein. However, while the link protein gene is known to reside on chromosome 5 in the human, the location of the aggrecan gene is currently undetermined in any species. The probe (pAGG2) for the aggrecan gene was mapped ...
Individuals with male factor infertility resulting from idiopathic oligo-, astheno- or teratozoospermia are frequently offered IVF in an attempt to increase their chances of having a child. A concern remains whether these infertile males have an elevated risk of transmitting chromosomal abnormalities to their offspring. Sperm chromosomal complements from these men were assayed using the human sperm/hamster oocyte fusion system and fluorescence in situ hybridization (FISH) on sperm nuclei. For each of 5 infertile patients, 100 sperm karyotypes were analyzed and multicolour FISH analysis was performed on a minimum of 10,000 sperm nuclei for each chromosome-specific DNAprobe for chromosomes 1 (pUC1.77), 12 (D12Z3), X (XC) and Y (DYZ3). As a group, the infertile patients showed increased frequencies of both numerical ({chi}{sup 2}=17.26, {proportional_to} <0.001) and total abnormalities ({chi}{sup 2}=7.78, {proportional_to} <0.01) relative ...
This paper described the application of an AC surface probe, similar to presently used eddy current probes, to the measurement of DC transport critical currents and critical state dissipation in high {Tc} superconductors. It has been shown that the probe can provide quantitative measurement of the full field penetration in superconducting samples by measuring the response of AC induced screening currents for superconducting materials in the form of tapes with overlayers of silver. In this manner, the AC probe can be used to replace the contact DC probe for determining critical currents in a noncontacting and local manner suitable for scanning over or along the sample.
This paper described the application of an AC surface probe, similar to presently used eddy current probes, to the measurement of DC transport critical currents and critical state dissipation in high T_c superconductors. It has been shown that the probe can provide quantitative measurement of the full field penetration in superconducting samples by measuring the response of AC induced screening currents for superconducting materials in the form of tapes with overlayers of silver. In this manner, the AC probe can be used to replace the contact DC probe for determining critical currents in a noncontacting and local manner suitable for scanning over or along the sample.
This paper described the application of an AC surface probe, similar to presently used eddy current probes, to the measurement of DC transport critical currents and critical state dissipation in high [Tc] superconductors. It has been shown that the probe can provide quantitative measurement of the full field penetration in superconducting samples by measuring the response of AC induced screening currents for superconducting materials in the form of tapes with overlayers of silver. In this manner, the AC probe can be used to replace the contact DC probe for determining critical currents in a noncontacting and local manner suitable for scanning over or along the sample.
It is both experimentally and theoretically demonstrated that ion flow velocity at an arbitrary angle with respect to the magnetic field can be measured with a directional Langmuir probe. Based on the symmetry argument, we show that the effect of magnetic field on directional probe current is exactly canceled in determining the ion flow velocity, and obtain the generalized relation between flow velocity and directional probe currents valid for any flowing direction. The absolute value of the flow velocity is determined by an in situ calibration method of the probe. The applicability limit of the present method to a strongly ion-magnetized plasma is experimentally examined. (author)
Excision repair in uv irradiated human fibroblasts has been examined in portions of DNA replicating after irradiation versus those remaining unreplicated. Two approaches, one using a uv-endonuclease to estimate pyrimidine dimers remaining in DNA, the other using density labeling to measure excision resynthesis, indicate that the extent of repair is the same for both replicated and unreplicated DNA.
Escherichia coli rnh mutants lacking RNase H activity are capable of recA+-dependent DNA replication in the absence of concomitant protein synthesis (stable DNA replication). In rnh dnaA::Tn10 and rnh...Full Text Available
In this paper the thermal equilibrium number of solitons in DNA as a function of absolute temperature and the number of base pairs is calculated. These calculations are effected by modeling DNA as a Toda lattice with parameters chosen to match experimentally measured properties of DNA. It is found that a significant number of solitons is generated at physiological temperature. 23 refs., 2 figs.
Transcriptional silencing in Saccharomyces cerevisiae occurs at several genetic loci, including the ribosomal DNA (rDNA). Silencing at telomeres (telomere position effect [TPE])...Full Text Available
We describe a method for affinity purification of sequence-specific DNA binding proteins that is fast and effective. Complementary chemically synthesized oligodeoxynucleotides that contain a recognition site for a sequence-specific DNA binding protein are annealed and ligated to give oligomers. This DNA is then covalently coupled to Sepharose CL-2B with cyanogen bromide to yield the affinity resin. A partially purified protein fraction is combined with competitor DNA and subsequently passed through the DNA-Sepharose resin. The desired sequence-specific DNA binding protein is purified because it preferentially binds to the recognition sites in the affinity resin rather than to the nonspecific competitor DNA in solution. For example, a protein fraction that is enriched for transcription factor Sp1 can be further purified 500- to 1000-fold by ...
A plethora of experimental studies use mtDNA as a marker of demographic processes without questioning the possibility that selection may bias their interpretations. We studied four lines of Drosophila melanogaster that have a standardized nuclear DNA but variable mtDNA. We completed the sequencing of the mitochondrial genomes (excluding the A+T rich region) and compiled the differences. We then assayed male influence on oviposition, starvation resistance, lipid proportion and physical activity. We discuss these results in terms of the known differences between the lines and conclude that naturally occurring mtDNA variants in D. melanogaster are expressed at the level of the organismal phenotype. PMID:21757031
We have designed a new nucleobase, benzodeazaadenine (BDA) that has a stronger charge transport ability than guanine and is not destroyed during charge transport process. By incorporating this new nucleobase into DNA, we demonstrated a protocol for real DNA nano-wire that is far superior to natural DNA. We also demonstrated that the selectivity for the interaction of Mn(II) ion with guanine N7 in G runs is a HOMO-controlled process, and as a consequence, the selectivity for G-metal ion interactions obtained by 15N-NMR studies would directly reflect the HOMO distribution of G-containing sequences in B-DNA. PMID:12903077
Computer programs have been developed for modelling impedance and transmit-receive eddy current probes in two-dimensional axis-symmetric configurations. These programs, which are based on analytic equations, simulate bobbin probes in infinitely long tubes and surface probes on plates. They calculate probe signal due to uniform variations in conductor thickness, resistivity and permeability. These signals depend on probe design and frequency. A finite element numerical program has been procured to calculate magnetic permeability in non-linear ferromagnetic materials. Permeability values from these calculations can be incorporated into the above analytic programs to predict signals from eddy current probes with permanent magnets in ferromagnetic tubes. These programs were used to test various probe designs for new testing applications. ...
Sentinel node (SN) scintigraphy for cervical and vulvar cancer guides the gynaecological oncologist in finding the metastatic lymph nodes during lymphadenectomy. The role of the surgical gamma probe in the sentinel node concept in gynaecological oncology is to localise (SN) both intra-operatively and transcutaneously. Intra-operative hand-held collimated gamma probes are increasingly used for detection of the sentinel lymph node. A comparative evaluation of handheld gamma probes: Neoprobe 1500, Europrobe, Gamma Finder, Gamma Ray Prospector GRP1 and GPR2 was performed using different detection methods. Laboratory tests were performed in which sensitivity, spatial resolution and angular sensitivity were evaluated. The results for each gamma probe were summarised and discussed. Awareness of a gamma probes capabilities and limitations should be considered in the appropriate selection of ...
Conventional eddy current bobbin probes are known to be ineffectual in detecting circumferential cracks in tubing. Multi-pancake and/or rotating pancake probes are required to detect circumferential cracks. It has recently been demonstrated in CANDU steam generator tubes, with deformation, ferromagnetic deposits, and copper deposits, that multi-channel probes with transmit-receive (T/R) coils are superior to those using surface impedance coils. Unlike rotating probes, the design of a new probe denoted as C3 permits fast, single-scan inspection of a full-length tube at inspection speeds comparable to conventional bobbin probes. Since 1992, the probe has been used routinely for steam generator in-service inspection at 4 CANDU plants. Defective tubes have been plugged and the units returned to service, and they continue to operate without ...
Conventional eddy current bobbin probes are known to be ineffectual in detecting circumferential cracks in tubing. Multi-pancake and/or rotating pancake probes are required to detect circumferential cracks. It has recently been demonstrated in CANDU steam generator tubes, with deformation, ferromagnetic deposits, and copper deposits, that multi-channel probes with transmit-receive (T/R) coils are superior to those using surface impedance coils. Unlike rotating probes, the design of a new probe denoted as C3 permits fast, single-scan inspection of a full-length tube at inspection speeds comparable to conventional bobbin probes. Since 1992, the probe has been used routinely for steam generator in-service inspection at 4 CANDU plants. Defective tubes have been plugged and the units returned to service, and they continue to operate without ...
Synthetic oligoribonucleotides have been used to probe the interaction of MS2 coat protein with the translational operator of the MS2 replicase gene. We have investigated the possible formation of a...Full Text Available
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
The introduction of prenatal screening requires rapid high-throughput diagnosis of common aneuploidies. Multiplex ligation-dependent probe amplification (MLPA) allows for quick, easily automated multiplex...Full Text Available
A new gadolinium chelating NIR fluorescent molecular probe increases T1 relaxivity of water protons, facilitating combined optical and magnetic resonance imaging.
Computer programs have been written to allow the analysis of different types of eddy-current probes and their performance under different steam generator test conditions. The probe types include the differential bobbin probe, the absolute bobbin probe, the pancake probe and the reflection probe. The generator test conditions include tube supports, copper deposits, magnetite deposits, denting, wastage, pitting, cracking and IGA. These studies are based mostly on computed values, with the limited number of test specimens available used to verify the computed results. The instrument readings were computed for a complete matrix of the different test conditions, and then the test conditions determined as a function of the readings by a least-squares technique. A comparison was made of the errors in fit and instrument drift for the different probe ...
CANDU steam generators and heat exchangers have been afflicted with a significant variety of degradation mechanisms. Until recently, detection and characterization of most of the varieties of flaws required usage of several different eddy current techniques. Tubes needed to be scanned multiple times to ensure they were inspected for all possible degradation mechanisms. The time required to deploy so many different types of techniques to inspect the steam generator tubes has had a significant impact on the inspection schedules. The X-Probe was designed with an array of eddy current coils capable of detecting, characterizing and sizing axial, circumferential and volumetric flaws. The X-Probe is a fast-scanning probe with scanning speed equivalent to speeds typically used for simple bobbin coil probes. With its capability of fulfilling most if not all of the functions of the other tube-testing eddy current ...
CANDU steam generators and heat exchangers have been afflicted with a significant variety of degradation mechanisms. Until recently, detection and characterization of most of the varieties of flaws required usage of several different eddy current techniques. Tubes needed to be scanned multiple times to ensure they were inspected for all possible degradation mechanisms. The time required to deploy so many different types of techniques to inspect the steam generator tubes has had a significant impact on the inspection schedules. The X-Probe was designed with an array of eddy current coils capable of detecting, characterizing and sizing axial, circumferential and volumetric flaws. The X-Probe is a fast-scanning probe with scanning speed equivalent to speeds typically used for simple bobbin coil probes. With its capability of fulfilling most if not all of the functions of the other tube-testing eddy current ...
This paper describes the technical design and performance of the diagnostic system installed in the tokamak TCABR to measure parameters of the plasma edge. The system consists of four Langmuir probes under remote control. (author)
The past decade has seen the development of eddy current probes for inspection of the mildly ferro-magnetic alloy Monel 400. Due to the rapid advances in permanent magnet technology similar probes have been upgraded to magnetically saturate, and hence ins...
The utility of a two-photon optical fiber fluorescence probe (TPOFF) for sensing and quantifying tumor fluorescent signals was tested in vivo. Xenograft tumors were developed in athymic mice using MCA207...Full Text Available
Synthetic complementary oligonucleotides are useful hybridization probes for the detection of mRNAs and genes encoding proteins for which only a partial amino acid sequence is known. Usually this involves...Full Text Available
The infection of cultured human cells with baboon endogenous virus (BEV) frequently leads to an association of viral DNA with a specific genetic locus (termed BEVI, for baboon endogenous virus infection) on chromosome 6. Restriction endonuclease digestion of DNA from BEV-infected human cells and their derived somatic cell clones frequently revealed a common cellular DNA sequence in the proximity of one of the junctions between cellular DNA and the integrated virus. We propose that a short cellular DNA sequence, repeated on chromosome 6 and separated by unique DNA sequences, presents a high-affinity target for the integration of BEV in human cells. PMID:6401843
The authors have cloned a full-length putative rat pancreatic lysophospholipase cDNA by an improved mRNA isolation method and cDNA cloning strategy using (/sup 32/P)-labelled nucleotides. These new methods allow the construction of a cDNA library from the adult rat pancreas in which the majority of recombinant clones contained complete sequences for the corresponding mRNAs. A previously recognized but unidentified long and relatively rare cDNA clone containing the entire sequence from the cap site at the 5' end to the poly(A) tail at the 3' end of the mRNA was isolated by single-step screening of the library. The size, amino acid composition, and the activity of the protein expressed in heterologous cells strongly suggest this mRNA codes for lysophospholipase.
The human mitochondrial DNA (mtDNA) is a closed circular, 16,569-bp double-stranded DNA, encoding 13 genes whose protein products are subunits of the oxidative phosphorylation system required for synthesis of most of the ATP consumed by eukaryotic cells. Point mutations of the mtDNA that cause multi-tissue, loss-of-energy syndromes, called mitochondrial encephalomyopathies (e.g., MERRF and MELAS), have been identified. In addition, large-scale deletions of the human mtDNA have been identified and are the molecular bases for the neonatal and adolescent onset loss-of-energy syndromes Pearson and Kearns-Sayer, respectively. 5 refs., 1 fig.
... The rounds were initiated by an exploding bridgewire detonator (EBW) and the ionisation probes were progressively cloud by the detonation front. ...
This paper describes a signal simulator which is designed to calibrate eddy current probes. By using simultaneously an excitation coil and an electrically conducting plane, the simulator can emulate signals which correspond to spot welds with different flaws. The presence of a conducting plane allows a wide variation of the amplitude and phase of the excitation coil voltage during the calibration of a probe. A mathematical model for the simulator is derived and used to study the variation of the impedance change upon the parameters of the probe. Numerical computation shows that the impedance change depends in an important way on the frequency of the excitation coil current.
Here the authors try to dissolve some commonly held misconceptions about the operation of eddy current probes used for inspecting heat exchanger tubes. With the help of computer model results, the authors demonstrate that, for optimized operation, absolute/differential probes should be operated with coil current flowing in phase with another. This month's feature should be of interest to researchers as well as for NDT field personnel.
Phased array provides many advantages over conventional ultrasonic testing method, but phased array has also limitations. This paper describes typical results of the experimental sound field analysis generated from the array probe. A photo-elastic ultrasonic visualization technique was applied in this study. The sound fields of shear wave generated from the array probe was equivalent to that from the fixed angle probe. (author)
To overcome the limitations of conventional bobbin style eddy-current probes, it has become common practice to perform some portion of a steam generator tube inspection program with array coil probes. Typically, eight surface-riding pancake coils are utilized in a probe head, either as eight independent coils (8 x 1) or in a parallel combination of four serially connected coils (4 x 4). The advantages of the array coils are improved sensitivity to intergranular attack (IGA), reduced interference from tube denting, and improved sensitivity to circumferential cracking. The current status of commercially available probes is such that several weaknesses in array coils have limited the widespread application of this technology. Task S 301-6 was conceived to address each of the weaknesses in order to produce a more reliable and cost-effective probe. The emphasis was on reliability, since ...
Heterochromatin constitutes a significant portion of the genome in higher eukaryotes; approximately 30% in Drosophila and human. Heterochromatin contains a high repeat DNA content and a low density of protein-encoding genes. In contrast, euchromatin is composed mostly of unique sequences and contains the majority of single-copy genes. Genetic and cytological studies demonstrated that heterochromatin exhibits regulatory roles in chromosome organization, centromere function and telomere protection. As an epigenetically regulated structure, heterochromatin formation is not defined by any DNA sequence consensus. Heterochromatin is characterized by its association with nucleosomes containing methylated-lysine 9 of histone H3 (H3K9me), heterochromatin protein 1 (HP1) that binds H3K9me, and Su(var)3-9, which methylates H3K9 and binds HP1. Heterochromatin formation and functions are influenced by HP1, Su(var)3-9, and the RNA interference (RNAi) ...
Werner syndrome (WS) is characterized by premature onset of age-associated disorders and predisposition to cancer. The WS protein, WRN, encodes 3′ → 5′ DNA helicase and 3′...Full Text Available
Cytosine methylation in DNA is a major epigenetic signal, and plays a central role in propagating chromatin status during cell division. However the mechanistic links between DNA methylation and histone methylation are poorly understood. A multi-domain protein UHRF1 (ubiquitin-like, containing PHD and RING finger domains 1) is required for DNA CpG maintenance methylation at replication forks, and mouse UHRF1-null cells show enhanced susceptibility to DNA replication arrest and DNA damaging agents. Recent data demonstrated that the SET and RING associated (SRA) domain of UHRF1 binds hemimethylated CpG and flips 5-methylcytosine out of the DNA helix, whereas its tandom tudor domain and PHD domain bind the tail of histone H3 in a highly methylation sensitive manner. We hypothesize that UHRF1 brings the two components (histones and DNA) carrying ...
DNA methylation of cytosine residues is a widespread phenomenon and has been implicated in a number of biological processes in both prokaryotes and eukaryotes. This methylation occurs at the 5-position...Full Text Available
BackgroundDuring the last ten years, major advances have been made in characterizing and understanding the evolution of mitochondrial DNA, the most popular marker of molecular biodiversity....Full Text Available
BackgroundTrypanosoma cruzi, the etiologic agent of Chagas Disease, is a major vector borne health problem in Latin America and an emerging...Full Text Available
We studied telomeric DNA in leukemic cells as well as in normal T cells, B cells, monocytes, polymorphonuclear leukocytes, and bone marrow hematopoietic progenitor cells. No marked differences were...Full Text Available
Digestion of plant chromatin from Brassica pekinensis and Matthiola incana with staphylococcus nuclease leads to a DNA repeat of 175 plus or minus 8 and a core size of 140 base pairs. DNase I digestion...Full Text Available
AbstractWe combined atomistic molecular-dynamics simulations with quantum-mechanical calculations to investigate the sequence dependence of the stretching behavior of duplex DNA. Our...Full Text Available
Human chromosomal fragile sites are specific loci that are especially susceptible to DNA breakage following conditions of partial replication stress. They often are found in genes involved in tumorigenesis...Full Text Available
We use high-density single nucleotide polymorphism (SNP) genotyping microarrays to demonstrate the ability to accurately and robustly determine whether individuals are in a complex genomic DNA mixture....Full Text Available
BackgroundThe Class II DNA transposons are mobile genetic elements that move DNA sequence from one position in the genome to another. We have previously demonstrated that the naturally...Full Text Available
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
Most codon indices used today are based on highly biased nonrandom usage of codons in coding regions. The background of a coding or noncoding DNA sequence, however, is fairly random, and can be characterized...Full Text Available
The sequence in the first hypervariable segment (HVS-I) of the control region has been used as a source of evolutionary information in most phylogenetic analyses of mtDNA. Population genetic inference...Full Text Available
BackgroundThe construction of cDNA libraries is a useful tool to understand gene expression in organisms under different conditions, but random sequencing of unbiased cDNA collections...Full Text Available
The integration sites for viral DNA in cells infected with Moloney murine leukemia virus (M-MuLV) were studied by restriction endonuclease cleavage of cellular DNA followed by electrophoresis in agarose...Full Text Available
The sequence-dependent structural variability and conformational dynamics of DNA play pivotal roles in many biological milieus, such as in the site-specific binding of transcription factors to target...Full Text Available
DNA interstrand cross-links (ICLs) are critical cytotoxic lesions produced by cancer chemotherapeutic agents such as the nitrogen mustards and platinum drugs; however, the exact mechanism of ICL-induced...Full Text Available
Because copper catalyzes the conversion of H2O2 to hydroxyl radicals in vitro, it has been proposed that oxidative DNA damage may be an important component of copper toxicity....Full Text Available
The bulky octahedral complex, Rh(bpy)2chrysi3+ (chrysi = 5, 6- chrysenequinone diimine), binds single base mismatches in a DNA duplex with micromolar binding affinities...Full Text Available
Auxin autonomous growth of most crown gall tumor cells requires the expression of two auxin biosynthesizing genes (tms 1 and tms 2) from the T-DNA of Agrobacterium...Full Text Available
The use of dideoxynucleotide triphosphates labeled with different fluorescent dyes (dye terminators) is the most versatile method for automated DNA sequencing. However, variation in peak heights reduces...Full Text Available
Chromosomal common fragile sites (CFSs) are genetically unstable regions of the genome that are induced by conditions that impair DNA replication. In this report, we show that treatment with the DNA...Full Text Available
Epigenetic remodeling is a hallmark of cancer, with the frequent acquisition of de novo DNA methylation in CpG islands. However, the functional relevance of de novo...Full Text Available
DNA double-strand breaks (DSBs) are generally accepted to be the most biologically significant lesion by which ionizing radiation causes cancer and hereditary disease. However, no information on the...Full Text Available
The development of chromatin immunoprecipitation methods coupled with DNA microarray (ChIP-chip) technology has enabled genome-wide identification of cis-DNA regulatory elements to which transcription...Full Text Available
DNA supercoiling in the chloroplast of the unicellular green alga Chlamydomonas reinhardtii was found to change with a diurnal rhythm in cells growing in alternating 12-h dark–12-h...Full Text Available
Investigations of enzymes involved in DNA metabolism have strongly benefited from the establishment of single molecule techniques. These experiments frequently require elaborate DNA substrates, which...Full Text Available
In the present study we have used the alkaline elution technique to study the effect of misonidazole (MISO) on the initial amount of DNA cross-linking in various normal and neoplastic tissues of C3H...Full Text Available
A number of aberrant morphological phenotypes were noted during propagation of the Arabidopsis thaliana DNA hypomethylation mutant, ddm1, by repeated self-pollination. Onset of a spectrum of morphological...Full Text Available
Caries is a multifactorial disease, and studies aiming to unravel the factors modulating its etiology must consider all known predisposing factors. One major factor is bacterial colonization,...Full Text Available
DNA replication in Escherichia coli cells lacking protein HU was studied. HU has been suggested to be involved in the initiation of replication from in vitro studies. The isolated HU mutants, however,...Full Text Available
ObjectiveThe present study was undertaken to analyze the impact of epigenetic alterations with a main focus on nuclear area, aneuploidy, hyperploidy, and proliferation...Full Text Available
The syntheses and applications of two metallointercalators, Rh(bpy)2(chrysi)3+ and Rh(bpy)2(phzi)3+, that target single base mismatches in DNA are...Full Text Available
A nonmammalian test system for germ-cell mutagenesis has been developed by using the Japanese medaka fish. We describe a system for detecting DNA alterations in F1 progeny descended from the gamma-irradiated...Full Text Available
The binding mode of telomestatin to G-quadruplex DNA has been investigated using electrospray mass spectrometry, by detecting the intact complexes formed in ammonium acetate. The mass measurements show...Full Text Available
Global hypomethylation has been shown to increase genome instability potentially leading to increased cancer risk. We determined whether global methylation in blood leukocyte DNA was associated...Full Text Available
DNA methylation is an important event in epigenetic changes in cells, and a fundamental regulator of gene transcription. Bisulfite genomic sequencing is a powerful technique used in studies of DNA methylation. However, the established procedures often require relatively large amounts of DNA. In everyday practice, samples submitted for analysis might contain very small amounts of poor quality material, as is often the case with forensic stain samples. In this study, we assess a modified, more efficient method of bisulfite genomic sequencing. Genomic DNA extracted from 3-mm dried blood spots using QIAamp micro kit was treated with sodium bisulfite (using EpiTect kit). Subsequent methylation-specific PCR (MSP) followed by DNA sequencing displayed the differentially methylated region of imprinted gene SNRPN. Our results show that this new combination of efficient ...
We recently reported that human immunodeficiency virus type 1 (HIV-1) unintegrated linear DNA displays a discontinuity in its plus strand, precisely defined by a second copy of the polypurine tract...Full Text Available
DNA antoantibodies in serum will bind to antigen-coated polystyrene tubes and can be detected by radiolabelled anti-immunoglobulin. The method is quantitative, gives information on the antibody class...Full Text Available
A cDNA encoding a cytochrome P450 enzyme was isolated from a cDNA library of the corpora allata (CA) from reproductively active Diploptera punctata cockroaches. This P450 from the endocrine...Full Text Available
Recent studies have indicated that the DNA replication machinery is coupled to silencing of mating-type loci in the budding yeast Saccharomyces cerevisiae, and a similar silencing mechanism...Full Text Available
Human BTEB cDNA clones have been isolated, sequenced, and the corresponding gene has been assigned to human chromosome 9, region q13, by fluorescent in situ hybridization and DNA blot analysis using DNAs from hybrid cell clones containing a single human chromosome. The cDNA clone encodes a polypeptide of 244 amino acids whose sequence shows a high sequence similarity with the rat BTEB (98% amino acid identity).
Until a decade ago, only the differential bobbin type eddy current probe was considered necessary for reliable heat exchanger tube inspection. The introduction of different tube materials, manufacturing processes and a variety of service induced failures has greatly increased the demands of eddy current testing. Optimized probe designs enhanced by improved instrumentation can help satisfy some of these demands. Some of the more difficult inspection problems are detection of circumferential cracks, fretting wear under non-ferromagnetic support plates and shallow internal defects. Reliable detection and sizing of such defects is often made more difficult by the fact they frequently occur in defect prone regions such as under tubesheets or support plates and in transition regions of finned tubes. Probe designs effective in overcoming these difficulties exist. This paper describes a number of such probes ...
A refined map for the linear arrangement of histones along DNA in nucleosomal core particles has been determined by DNA-protein crosslinking. On one strand of 145-bp core DNA, histones are aligned in...Full Text Available
Two cDNA libraries corresponding to polyA+ RNA from human adult skeletal muscle have been constructed by cloning in the PstI site of pBR322. Skeletal alpha actin cDNA clones have been isolated and characterized....Full Text Available
Ditercalinium, an antitumor bifunctional intercalator which forms a high affinity reversible complex with DNA, was found to be specifically cytotoxic for polA and lig7 E. coli strains. In the polA strain,...Full Text Available
Fission yeast S. pombe is assumed to be a good model for cloning of human DNA repair genes, because human gene is normally expressed in S. pombe and has a very similar protein sequence to yeast protein. We have tried to elucidate the DNA repair mechanisms of S. pombe as a model system for those of mammals. (J.P.N.)
Two cellular DNA regions representing common domains for proviral DNA integration ( Mlvi -1 and Mlvi -2) have been identified in Moloney murine leukemia virus-induced rat thymic lymphomas. Cellular...Full Text Available
DNA isolation procedures significantly influence the outcome of PCR-based detection of human pathogens. Unlike clinical samples, DNA isolation from food samples such as fresh and fresh-cut produce has remained a formidable task and has hampered the sensitivity and accuracy of molecular methods. We...
We have examined homologous fragments of DNA cloned from two different tissues for changes in the dNA sequence which might be related to tissue specific gene expression. The 5' end of the chicken ovalbumin...Full Text Available
This paper presents a mathematical model that calculates eddy current signals from laterally spaced transmit-receive pancake coils. This model was validated by comparing calculations with laboratory measurements and it has been applied to evaluating design variations in X-Probe eddy current array coils. This model was used to quantify detection capabilities of probe designs with different numbers of coils. The current standard X-Probe design for 13 mm (0.5 inch) diameter steam generator tubes uses 3 rows of coils with 8 coils per row. This design is commonly referred to as the '8 coil X-Probe'. A significant improvement in detection capability was predicted for an X-Probe with 12 coils per row (called the '12 coil X-Probe') detecting short circumferential crack-like flaws. Based on these calculations, a prototype 12 coil ...
The bobbin probe technique is basically one of the important ECT methods for the steam generator tube integrity assessment that is practiced during each plant outage. The bobbin probe also is the essential component which consists of the whole ECT examination system, and provides a decisive data for the evaluation of tube integrity in compliance with acceptance criteria described in specific procedures. The selection of probe is especially important because the quality of acquired ECT data is determined by the probe design characteristics, such as geometry and operation frequency, and has an important effect on examination results. The Electric Power Research Institute (EPRI) has recently defined the procedures for the qualification of eddy current hardware and technique. These procedures provide two basic methods for qualification. Flawed tube removed from operation, or artificial flaw is required for ...
The bobbin probe technique is basically one of the important ECT methods for the steam generator tube integrity assessment that is practiced during each plant outage. The bobbin probe also is the essential component which consists of the whole ECT examination system, and provides a decisive data for the evaluation of tube integrity in compliance with acceptance criteria described in specific procedures. The selection of probe is especially important because the quality of acquired ECT data is determined by the probe design characteristics, such as geometry and operation frequency, and has an important effect on examination results. The Electric Power Research Institute (EPRI) has recently defined the procedures for the qualification of eddy current hardware and technique. These procedures provide two basic methods for qualification. Flawed tube removed from operation, or artificial flaw is required for ...
Helically coiled tubes of steam generators (SG) in FBR are boundaries between sodium and water/steam. Therefore, to assure the integrity of tubes, it is necessary to inspect the tubes nondestructively for in service or after a sodium-water reaction accident. In order to make it possible to conduct in-service inspection of SG tubes, we have studied on eddy current probes and probe inserting methods. As for the probe inserting method, IHI designed a fluid driving type which consists of a model probe and signal cable with float balls and driven by air pressure force. Presented in this paper is the authors' report, which describes the fluid driving type as an effective method to insert an eddy current probe into helically coiled tubes. The outline of the test results is as follows: 1. It was possible to insert the probe into 65 meter length helically coiled tubes. ...
Five corrosion probes were received from West Valley Nuclear Services for evaluation in simulated tank 8D-2 3rd-stage sludge wash slurry. The same waste slurry simulated was also used in a series of ongoing corrosion studies assessing the effects of in-tank sludge washing on the integrity of tank 8D-2. Two of the corrosion probes were installed in the coupon corrosion test vessels operating at {approximately}150{degrees}F to compare performance of the probes with that observed by coupon tests conducted in the same vessels. Corrosion rate data calculated from electrical resistance measurements of the corrosion probes were evaluated for this study using two slightly different approaches. One approach uses the total length of exposure of the probe to give a ``time-averaged`` value of the corrosion rate. The other approach uses a shorter period of time (relative to the length of the ...
Steam Generator (SG) tubes in PWR plant are periodically inspected using eddy current testing (ECT) method. In Japan, bobbin coil type dual function probe is used for full length inspection, but the detectability for circumferential crack is very poor, and to improve detectability for circumferential crack and supplement regular ECT, various kinds of multi segment eddy current probes and multi transducer ultrasonic testing (UT) method have been developed. ECT probes are constructed of multiple surfaceriding special winding differential type coils and UT probe consists of 16 transducers which are spaced equally around the probe circumference for 360deg wall coverage. Both of those EC and UT probes are designed effective to detect circumferential crack. On the other hand, various kinds of laboratory induced circumferential cracks have been made and destroyed to ...
The Electromagnetic Fluctuations (EMF) diagnostic will be used to monitor ion fluctuations which could be unstable in MFTF-B. Each probe assembly includes a high impedance electrostatic probe to measure potential fluctuations, and a group of nested, single turn loops to measure magnetic fluctuations in three directions. Eventually, more probes and loops will be added to each probe assembly for making more detailed measurements. The sensors must lie physically close to the plasma edge and are radially positionable. Also, probes at separate axial locations can be positioned to connect along the same magnetic field line. These probes are similar in concept to the rf probes used on TMX, but the high thermal load for 30-second shots on MFTF-B requires a water-cooled design along with temperature monitors. Each signal channel has a bandwidth of ...
In this Letter, we demonstrate the application of time-resolved fluorescence anisotropy measurements to detect solution state hybridization of streptavidin conjugate (CdSe)ZnS quantum dots (QD). The study was performed on samples containing 10nM QD incubated with 800nM DNA. We show that the rotational correlation time of QD-DNA constructs increases significantly upon hybridization with values of 330ns (QD-ssDNA) and 1.3ms (QD-dsDNA), corresponding to a diameter of 14nm and 23nm respectively. The present study opens a new modality for hybridization detection using quantum dots.
Lebers hereditary optic neuropathy (LHON), the most frequent mitochondrial disorder, is mostly due to three mitochondrial DNA (mtDNA) mutations in respiratory chain complex I subunit genes: 3460/ND1, 11778/ND4 and 14484/ND6. Despite considerable clinical evidences, a genetic modifying role of the mtDNA haplogroup background in the clinical expression of LHON remains experimentally unproven. We investigated the effect of mtDNA haplogroups on the assembly of oxidative phosphorylation (OXPHOS) complexes in transmitochondrial hybrids (cybrids) harboring the three common LHON mutations. The steady-state levels of respiratory chain complexes appeared normal in mutant cybrids. However, an accumulation of low molecular weight subcomplexes suggested a complex I assembly/stability defect, which was ...
Since the elucidation of the structure of double helical DNA, the construction of small molecules that recognize and react at specific DNA sites has been an area of considerable interest. In particular, the study of transition metal complexes that bind DNA with specificity has been a burgeoning field. This growth has been due in large part to the useful properties of metal complexes, which possess a wide array of photophysical properties and allow for the modular assembly of an ensemble of recognition elements. Here we review recent experiments in our laboratory aimed at the design and study of octahedral metal complexes that bind DNA non-covalently and target reactions to specific sites. Emphasis is placed both on the variety of methods employed to confer site-specificity and upon the many applications for these complexes. Particular attention is given to the family of complexes recently designed that ...
Since the elucidation of the structure of double helical DNA, the construction of small molecules that recognize and react at specific DNA sites has been an area of considerable interest. In particular, the study of transition metal complexes that bind DNA with specificity has been a burgeoning field. This growth has been due in large part to the useful properties of metal complexes, which possess a wide array of photophysical attributes and allow for the modular assembly of an ensemble of recognition elements. Here we review recent experiments in our laboratory aimed at the design and study of octahedral metal complexes that bind DNA non-covalently and target reactions to specific sites. Emphasis is placed both on the variety of methods employed to confer site-specificity and upon the many applications for these complexes. Particular attention is given to the family of complexes recently designed that ...
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 ...
The glass gene is required for proper photo-receptor differentiation during development of the Drosophila eye glass codes for a DNA-binding protein containing five zinc fingers that we show is a transcriptional activator. A comparison of the sequences of the glass genes from two species of Drosophila and a detailed functional domain analysis of the Drosophila melanogaster glass gene reveal that both the DNA-binding domain and the transcriptional-activation domain are highly conserved between the two species. Analysis of the DNA-binding domain of glass indicates that the three carboxyl-terminal zinc fingers alone are necessary and sufficient for DNA binding. We also show that a deletion mutant of glass containing only the DNA-binding domain can behave in a dominant-negative manner both in vivo and in a cell culture assay that measures transcriptional activation. PMID:7604032
In evaluating the risk associated with low levels of exposure to toxicants, it is clear that DNA repair, one of the main defenses against agent damage, is not a constant. It can be modified by age, time of day, and physiological state. Nutrition, especially caloric restriction (CR), can modify almost every step in the process of protecting genomic integrity. And history of exposure can modify DNA repair. Thus, the conditions of exposure are almost as important to toxicity as the exposure itself, even at the level of DNA repair. Extrapolation from high to low dose, to be consistent with what is known, should be less a mathematical exercise than an exercise in toxicological judgement, which puts the exposure in proper perspective. This appears to be true at almost every level in the process including a response with a toxic stimulus, even those thought to be very basic, such as DNA repair.
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...
We propose a combined experimental (Atomic Force Microscopy) and theoretical study of the structural and dynamical properties of nucleosomes. In contrast to biochemical approaches, this method allows to determine simultaneously the DNA complexed length distribution and nucleosome position in various contexts. First, we show that differences in the nucleo-proteic structure observed between conventional H2A and H2A.Bbd variant nucleosomes induce quantitative changes in the in the length distribution of DNA complexed with histones. Then, the sliding action of remodeling complex SWI/SNF is characterized through the evolution of the nucleosome position and wrapped DNA length mapping. Using a linear energetic model for the distribution of DNA complexed length, we extract the net wrapping energy of DNA onto the histone octamer, and compare it to previous studies.
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 ...
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 ...
The parmbsc0 force field was applied to study in detail the binding of netropsin, at a salt concentration of 0.28M Na^+, to the minor groove of an 8-mer (5'CCAATTGG)2 DNA duplex forming a netropsin.DNA complex which previously has been characterized by X-ray crystallography, albeit with the use of closely related DNA duplexes. The X-ray structure revealed that the terminal guanidinium and amidinium groups of netropsin interact with the extreme ends of the palindromic AATT sequence of the receptor DNA. The parmbsc0 parameters of B-DNA and AMBER v9 parameters of netropsin generated a stable 6ns molecular dynamics (MD) trajectory for a 1:1 class I binding motif of this complex. Trajectory analysis for the salt and hydration effects on the binding of netropsin to the 8-mer DNA duplex revealed ...
DNA polymerase gamma from purified nuclei of EMT-6 cells (mice) seems to be identical to the mitochondrial DNA polymerase from the same source following several criteria. These two enzyme activities are strongly inhibited by ethidium bromide and acriflavin, while proflavin, acridine orange, daunomycin and chloroquine inhibition is less pronounced. In the case of DNA polymerases alpha and beta very little inhibition by ethidium bromide was observed. Intercalation of this dye in a poly dA-dT 12-18 template-primer was studied spectrophotometrically under conditions similar to those in the in vitro DNA polymerase assay. The polymerase assay. The inhibition by this drug of the mitochondrial DNA polymerase gamma activity was shown to be competitive at varying concentrations of TTP while the inhibition was of the non-competitive type at different concentrations of poly dA-dT 12-18. We ...
Radioadapted chicken embryo cells (X-irradiation in ovo with 10 cGy at the 14th day of development with priming periods of 24 h) were treated in vitro by challenge doses of 14 different DNA- and/or chromatin-interactive agents, including X-rays. A decrease in the cellular damage, as measured by scheduled DNA synthesis, was only observed with X-irradiation. Sedimentation of nucleoids as well as viscosity of alkaline lysates from ethidium bromide (0.35-400 {mu}g/ml)-, vovobiocin (125-1800 {mu}g/ml)-, and hyperthermia (30 min at 43 and 45 )-treated cells suggest a higher tendency of radioadapted cells to undergo positive DNA supercoiling. When DNA from adapted and non-adapted chicken embryo cells was used as substrate, neither its digestion by DNase I nor the inhibition of DNase I activity by various DNA-interactive agents was changed in primed cells. From the previous investigations ...
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 ...
Bacteriophage M13 mp10 DNA were irradiated with near-UV light in the presence of tetracycline derivatives and primed with synthetic oligonucleotide to be used for DNA synthesis using Escherichia coli DNA polymerase. Chain terminations were observed by denaturing polyacrylamide gel electrophoresis and mapped precisely. All the synthesis stops occurred before or at the level of guanine residues, showing that the photoreaction mediated by tetracycline derivatives led to a preferential alteration of guanine residues. These lesions were demonstrated to be induced in DNA through a pathway involving singlet oxygen. Tetracycline derivatives also photoinduced the breakage of the DNA sugar-phosphate backbone monitored by the conversion of supercoiled phi X174 DNA to a relaxed form. This lesion was shown to be initiated by hydroxyl radicals. The production of this free ...
In the last two decades, due to increased inspection demands, eddy current instrumentation has advanced from single-frequency, single-output instruments to multifrequency, computer-aided systems. This has significantly increased the scope of eddy current testing, but, unfortunately, it has also increased the cost and complexity of inspections. In addition, this approach has not always improved defect detectability or signal-to-noise. Most eddy current testing applications are still performed with impedance probes, which have well known limitations. However, recent research at AECL has led to improved eddy current inspections through the design and development of transmit-receive (T/R) probes. T/R eddy current probes, with laterally displaced transmit and receive coils, present a number of advantages over impedance probes. They have improved signal-to-noise ratio in the presence of variable lift-off ...
Bobbin-coil-type eddy current probes, which are conventionally used for nondestructive inspection of steam generator tubes in pressurized-water-type nuclear power plants, have poor detectability for circumferential flaws. Hence a new type of eddy current probe was proposed to detect effectively the magnetic flux component disturbed by a flaw and thus to eliminate the flaw direction dependency on the flaw detectability. In the course of development of the proposed method, structures of the probe were investigated based on the measurement of magnetic fields induced by exciting flat coils with several shapes. The new type of probe proposed here consists of differential pick-up coils detecting magnetic flux and exciting coils having a parallelogrammic shape, and its structure was fabricated experimentally in order to detect flaws independently of their directions. Nondestructive flaw detection tests was ...
Although the p53 tumor suppressor is relatively well characterized, much less is known about the functions of other members of the p53 family, p73 and p63. Here, we present evidence that in specific pathological conditions caused by exposure of normal cells to bile acids in acidic conditions, p73 protein plays the predominant role in the DNA damage response. These pathological conditions frequently occur during gastric reflux in the human esophagus and are associated with progression to esophageal adenocarcinoma. We found that despite strong DNA damage induced by bile acid exposure, only p73 (but not p53 and p63) is selectively activated in a c-Abl kinase-dependent manner. The activated p73 protein induces DNA damage repair. Using a human DNA repair PCR array, we identified multiple DNA repair genes affected by p73. Two glycosylases involved in base excision repair, SMUG1 and MUTYH, ...
Full text: DNA rearrangement events leading to chromosomal aberrations are central to ionizing radiation-induced cell death. Although DNA double-strand breaks are probably the lesion that initiates formation of chromosomal aberrations, little is understood about the molecular mechanisms that generate and modulate DNA rearrangement. Examination of the sequences that flank sites of DNA rearrangement may provide information regarding the processes and enzymes involved in rearrangement events. Accordingly, we developed a method using inverse PCR that allows the detection and sequencing of putative radiation-induced DNA rearrangements in defined regions of the human genome. The method can detect single copies of a rearrangement event that has occurred in a particular region of the genome and, therefore, DNA rearrangement detection does not require survival and ...
Electrochemical corrosion rate probes were constructed and tested along with mass loss coupons in a N2/O2/CO2 plus water vapor environment. Temperatures ranged from 450 to 600 C. Corrosion rates for ash-covered mild steel, 304L SS, and 316L SS probes using electrochemical techniques were a function of time, temperature, and process environment. Correlation between electrochemical and mass loss corrosion rates was good.
This paper reports on the electron current to a Langmuir probe in a moving high-pressure plasma. The plasma is an atmospheric-pressure propane flame, and the probe's speed relative to the plasma is varied from 10 to 30 ms"-"1. The current is linearly dependent on the speed and appears to be comprised of two components - one driven by diffusion and the other by convection. A model is devised which can be used to measure the electron density. By comparing the density computed from this model to that measured from the ion current, it has been possible to measure the electron mobility in the flame gases. The value obtained here is in good agreement with theoretical calculations found in the literature.
The paper is concerned with the simulation of practical testing situations which are too difficult and/or expensive to replicate in a laboratory environment. Numerical experiments are described which simulate the differential eddy current probe response to the build-up and chemical flushing of magnetite in the crevice gap of a PWR steam generator unit. The simulation results agree well with the only experimental data available to the authors and lead to the conclusion that conventional differential eddy current probes should be capable of characterizing crevice gap conditions with respect to the presence of magnetite. (author).
We consider a D-brane probe in the unstable string background associated with flux branes. The twist in the spacetime metric reponsible for supersymmetry breaking is shown to manifest itself in the mixing of open Wilson lines with the phases of some adjoint matter fields, resulting in a nonlocal and nonsupersymmetric form of Yang-Mills theory as the probe dynamics. This provides a setup where one can study the fate of a large class of unstable closed string theories that includes as a limit type 0 theories and various orbifolds of type II and type 0 theories. We discuss the limit of C/Z_n orbifold in some detail and speculate on the couplings with closed string tachyons.
"ID# ","Measurement ID Number " "DATE ","Date " "JDATE ","Julian Date " "TIME ", "Local Time " "GMT ","Greenwich Mean Time " "BAND ","Dielectric Probe Band ...
... hypersonic aerodynamic force, or the space-equivalent region. Some of these experiments change the X-15 from a research airplane to a kind of space probe, ...
In this paper, four different types of pulsed eddy current (PEC) probe are designed and their performance of detecting wall thickness reduction is compared. By using the backward difference method in time and the finite element method in space, PEC signals from various thickness and materials are numerically calculated and three features of the signal are selected. Since PEC signals and features are obtained by various types and sizes of probe, the comparison is made through the normalized features which reflect the sensitivity of the feature to thickness reduction. The normalized features indicate that the shielded reflection probe provides the best sensitivity to wall thickness reduction for all three signal features. Results show that the best sensitivity to thickness reduction is achieved by the peak value, but also suggest that the time to peak can be a good candidate because of its linear relationship with the ...
With the improvements accomplished during the past 15 years in detection techniques and instrumentation and with the opening of space exploration, molecular spectroscopy has become a very efficient way to probe planetary atmospheres.
Computer programs have been written to allow the analysis of different types of eddy-current probes and their performance under different steam generators test conditions. The probe types include the differential bobbin probe, the absolute bobbin probe, the pancake probe and the reflection probe. The generator test conditions include tube supports, copper deposits, magnetite deposits, denting, wastage, pitting, cracking, and intergranular attack. These studies are based mostly on computed values, with the limited number of test specimens available used to verify the computed results. The instrument readings were computed for a complete matrix of the different test conditions, and then the test conditions determined as a function of the readings by a least-squares technique. A comparison was made of the errors in fit and instrument drift for the different ...
... A second hydrometeor sampler (MRI Model 1220 ... The cloud particle probe normally installed in ... to the northeast showed no significant movement. ...
Integration of Eddy Current probes with MAUS scanning system has been demonstrated to produce scan rates of 36 sqft/hour. System will be designed to ...
Briefly, 48 MOS microparticle sensors (essentially the same as those flown on .... at the Naval Research Lab to interpret the polarization of the incoming radio .... Adapter Satellite, to be published in Journal of Spacecraft and Rockets, 1994. ...
... perform the inspection in the extremely limited space between adjacent blades and at the blade root, miniature eddy current probes were developed ...
Radixin is a cytoskeletal protein that may be important in linking actin to the plasma membrane. Recent cloning of the murine and porcine radixin cDNAs revealed a protein highly homologous to ezrin and moesin. The authors have cloned and sequenced the human radixin cDNA and found the predicted amino acid sequence for the human protein to be nearly identical to those predicted for radixin in the two other species. By Southern analyses of Chinese hamster x human somatic cell hybrid DNA and of PCR products derived from hybrids, the coding gene (RDX) was mapped to 11q. Fluorescence chromosomal in situ hybridization with a cDNA plasmid further localized this gene to band 11q23. However, PCR amplification with [open quotes]radixin-specific[close quotes] primers on the hybrid DNA panel yielded an additional, very similar DNA sequence that was further characterized by direct sequencing of ...
The overall objective of this project has been to develop immunochemical methods to quantitate unique DNA base damages in order to facilitate studies on radiation-induced damage production and repair. Specifically, we have been using antibodies raised to damaged bases to quantitate unique lesions in model systems in order to evaluate their potential biological consequences. Our approach has been to synthesize modified nucleotides or nucleosides, conjugate them to protein carriers, and use the conjugates as immunogens in rabbits or to prepare monoclonal antibodies. We have been studying damages that are stable radiolysis products found in X-irradiated DNA and thus of potential biological consequence. Our aim is to build an in vitro and in vivo data base on the interactions between model DNA lesions and such cellular enzymes as DNA polymerases and repair endonucleases. Initial studies have focused on ...
The overall objective of this project has been to develop immunochemical methods to quantitate unique DNA base damages in order to facilitate studies on radiation-induced damage production and repair. Specifically, we have been using antibodies raised to damaged bases to quantitate unique lesions in model systems in order to evaluate their potential biological consequences. Our approach has been to synthesize modified nucleotides or nucleosides, conjugate them to protein carriers, and use the conjugates as immunogens in rabbits or to prepare monoclonal antibodies. We have been studying damages that are stable radiolysis products found in X-irradiated DNA and thus of potential biological consequence. Our aim is to build an in vitro and in vivo data base on the interactions between model DNA lesions and such cellular enzymes as DNA polymerases and repair endonucleases. Initial studies have focused on ...
Summary DNA topoisomerases are the key enzymes involved in carrying out high precision DNA transactions inside the cells. However, they are detrimental to the cell when a wide variety of topoisomerase-targeted drugs generate cytotoxic lesions by trapping the enzymes in covalent complexes on the DNA. The discovery of unusual heterodimeric topoisomerase I in kinetoplastid family added a new twist in topoisomerase research related to evolution, functional conservation and their preferential sensitivity to Camptothecin. On the other hand, structural and mechanistic studies on kinetoplastid topoisomerase II delineate some distinguishing features that differentiate the parasitic enzyme from its prokaryotic and eukaryotic counterparts. This review summarizes the recent advances in research in kin...
We present the synthesis, magnetic and UV spectrometry of NiZn-ferrite nanofiber. The single phase of spinel ferrite was obtained at 600 {sup o}C. The NiZn-ferrite fibers fabricated by an electrospinning process were formed as a polygonal grain growth with firing temperature in fiber matrix. It appeared that the saturation magnetization (M{sub S}) of NiZn-ferrite nanofiber was dependent on Ni/Zn molar ratio which is similar to that of the inverse spinel ferrites. The NiZn-ferrite fibers showed good DNA adsorption efficiency that can be modified and utilized for DNA separation with magnetic nanofiber as a novel material in clinical applications.
E. coli HB101 cells transformed to tetracycline resistance with the plasmids pMB9 or pBR322 display a 10/sup 5/-10/sup 6/-fold lower plating efficiency on agar containing 440 ..mu..M CdCl/sub 2/ than nontransformed cells. When DNA is inserted into the BamH1 site of the plasmid tet gene, or when DNA spanning the BamH1 site is deleted, tetracycline resistance and cadmium hypersensitivity are both lost. In contrast, insertion of DNA into the ampicillin resistance gene does not affect cadmium hypersensitivity.
Nanoparticles represent versatile building blocks in material science and nanotechnology. Thereby, the defined assembly of nanostructures (13 and 56 nm in diameter, respectively) is of significant importance. Short DNA sequences can be bound to the nanoparticle surface thus enabling highly specific DNA hybridization-driven events that direct the formation of nanoparticle constructs.In this paper, examples for the defined formation of gold nanoparticle constructs are demonstrated. In addition, gold-silver core-shell nanoparticles are introduced as further building blocks for the hybridization-controlled formation of nanoparticle constructs.
Studies begun during the first year of this project on biopsies from spontaneous tumors in dogs and from human patients undergoing radiotherapy treatment were continued. The results indicate that significant fractions of the spontaneous tumors in dogs and cervical carcinomas in patients undergoing radiotherapy have a DNA content higher than normal diploid cells. The higher DNA content in tumor cells permits distinguishing them from normal cells in the biopsy material with the aid of flow microfluorometric (FMF) instrumentation or multiangle light-scatter spectrometry.
A colony PCR technique was applied for both genomic and chloroplast DNA in the green microalgae Chlorella. Of five different lysis buffers, Chelex-100 was superior for DNA extraction, PCR and DNA storage. It also was insensitive to variations in cell density. The conditions established for an improved PCR formulation are applicable for screening of genetically-engineered transformants as well as bioprospecting of natural microalgal isolates. Besides multiple Chlorella species, we also demonstrate the efficacy of Chelex-100 for colony PCR with a number of other microalgal strains, including Chlamydomonas reinhardtii, Dunaliella salina, Nannochloropsis sp., Coccomyxa sp., and Thalassiosira pseudonana. PMID:21431847
This paper presents the concept of a biological inspired reconfigurable hardware cell architecture which supports self-organisation and self-healing. Two fundamental processes in biology, namely fertilization-to-birth and cell self-healing have inspired the development of this cell architecture. In biology as well as in our hardware cell architecture it is the DNA which enables these processes. We propose a platform based on the electronic DNA (eDNA) and show through simulation, its capabilities as a new generation of robust reconfigurable hardware platforms. We have created a Java based simulator to simulate our self-organisation and self-healing algorithms and the results obtained from this looks promising.
Swollen basiodiospores of an adenine auxotroph of Phanerochaete chrysosporium were protoplasted with Novozyme 234 and transformed to prototrophy by using a plasmid containing the gene for an adenine biosynthetic enzyme from Schizophyllum commune. Transformation frequencies of 100 transformants per {mu}g of DNA were obtained. Southern blot analysis of DNA extracted from transformants demonstrated that plasmid DNA was integrated into the chromosomal DNA in multiple tandem copies. Analysis of conidia and basiodiospores from transformants demonstrated that the transforming character was mitotically and meiotically stable on both selective and nonselective media. Genetic crosses between double mutants transformed for adenine prototrophy and other auxotrophic strains yielded Ade{sup {minus}} progeny, which indicated that integration occurred at a site(s) other than the resident adenine biosynthetic gene.
Mitochondrial dysfunction has been implicated in premature aging, age-related diseases, and tumor initiation and progression. Alterations of the mitochondrial genome accumulate both in aging tissue...Full Text Available
In an effort to develop octahedral metal complexes as chemotherapeutic and diagnostic agents targeted to DNA, it is critical to optimize the properties of their cellular uptake. Appending d-octaarginine...Full Text Available
High-throughput technologies for DNA sequencing and for analyses of transcriptomes, proteomes and metabolomes have provided the foundations for deciphering the structure, variation and function of the...Full Text Available
The penetrance of Lebers hereditary optic neuropathy (LHON) in families with primary mitochondrial DNA (mtDNA) mutations is very complex. Matrilineal and nuclear genetic background, as well as environmental factors, have been reported to be involved in different affected pedigrees. Here we describe two large Chinese families that show a striking difference in the penetrance of LHON, in which 53.3% and 15.0% of members were affected (Pin vivo mitochondrial oxidative metabolism as described in a previous study. Evolutionary analysis failed to indicate any putatively pathogenic mutation that cosegregated with G11778A in these two pedigrees. Our results suggest that the variable penetrance of LHON in the two Chinese families is independent of both their mtDNA haplotype background and a seconda...
Senescence is regarded as a physiological response of cells to stress, including telomere dysfunction, aberrant oncogenic activation, DNA damage, and oxidative stress. This stress response has an antagonistically...Full Text Available
Polycyclic aromatic hydrocarbons (PAHs) are combustion products of organic materials, mixtures of which contain multiple known and probable human carcinogens. PAHs occur in indoor and outdoor air, as well as in char-broiled meats and fish. Human exposure to PAHs occurs by inhalation, ingestion and topical absorption, and subsequently formed metabolites are either rendered hydrophilic and excreted, or bioactivated and bound to cellular macromolecules. The formation of PAH-DNA adducts (DNA binding products), considered a necessary step in PAH-initiated carcinogenesis, has been widely studied in experimental models and has been documented in human tissues. This review describes immunohistochemistry (IHC) studies, which reveal localization of PAH-DNA adducts in human tissues, and semi-quantify PAH-DNA adduct levels using the Automated Cellular Imaging System (ACIS). These studies have shown that ...
BackgroundSection Calochroi is one of the most species-rich lineages in the genus Cortinarius (Agaricales, Basidiomycota) and is widely distributed...Full Text Available
This paper describes the search for new potential chemotherapeutic agents based on transition metal complexes with planar ligands. In this study, palladium polypyridyl complexes were synthesized and characterized by elemental analysis, NMR, UV-VIS and IR spectroscopies. The interaction of the complexes with DNA was also investigated by spectroscopic methods. All metal-to-ligand charge transfer (MLCT) bands of the palladium polypyridyl complexes exhibited hypochromism and red shift in the presence of DNA. The binding constant and viscosity data suggested that the complexes [PdCl{sub 2}(phen)] and [PdCl{sub 2}(phendiamine)] interact with DNA by electrostatic forces. Additionally, these complexes induced an important leishmanistatic effect on L. (L.) mexicana promastigotes at the final concentration of 10 {mu}mol L{sup -1} in 48 h. (author)
BackgroundGenetic predisposition to scrapie in sheep is associated with several variations in the peptide sequence of the prion protein gene (PRNP). DNA-based tests...Full Text Available
ABSTRACT. Chlorarachniophytes are enigmatic marine unicellular algae that acquired photosynthesis by secondary endosymbiosis. Chlorarachniophytes are unusual in that the nucleus of the engulfed algal cell (a green alga) persists in a miniaturized form, termed a nucleomorph. The nucleomorph genome of the model chlorarachniophyte, Bigelowiella natans CCMP621, is 373 kilobase pairs (kbp) in size, the smallest nuclear genome characterized to date. The B. natans nucleomorph genome is composed of three chromosomes, each with canonical eukaryotic telomeres and sub telomeric ribosomal DNA (rDNA) operons transcribed away from the chromosome end. Here we present the complete rDNA operon and telomeric region from the nucleomorph genome of Lotharella oceanica CCMP622, a newly characterized chlorarachn...
Pork identification in four types of food products, which are sausages and the casings, bread and biscuits, using species-specific polymerase chain reaction (PCR) detection of a conserved region in the mitochondrial (mt) 12S ribosomal RNA (rRNA) gene was developed. Genomic DNA of the food products were successfully extracted except for the casing samples, where no genomic DNA was detected. The extracted genomic DNA was then subjected to PCR amplification targeting the specific regions of the 12S rRNA gene. The genomic DNA from the food products were found to be of good quality and produced clear PCR products on the amplification of 12S rRNA gene of 387 base pairs (bp) from pork species. The species-specific PCR identification yielded excellent results for identification of pork derivatives...
We explored transcriptional responses of the fission yeast Schizosaccharomyces pombe to various environmental stresses. DNA microarrays were used to characterize changes in expression...Full Text Available
Motivation: DNA sequences can be represented by sequences of four symbols, but it is often useful to convert the symbols into real or complex numbers for further analysis. Several mapping...Full Text Available
By assuming that not only counter-ions but DNA molecules as well are thermally distributed according to a Boltzmann law, we propose a modified Poisson-Boltzmann equation, at the classical level, as a starting point to compute the effects of quantum fluctuations of the electric field on the interaction among DNA-cation complexes. The latter are modeled here as infinite one-dimensional wires (?-functions). Our goal is to single out such quantum-vacuum-driven interaction from the counterion-induced and water-related interactions. We obtain a universal, frustration-free Casimir-like (codimension 2) interaction that extensive numerical analysis show to be a good candidate to explain the formation and stability of DNA aggregates. Such Casimir energy is computed for a variety of configurations of...
As adulterated and substituted Chinese medicinal materials are common in the market, therapeutic effectiveness of such materials cannot be guaranteed. Identification at species-, strain- and locality-levels,...Full Text Available
Curcumin, the yellow pigment from the rhizoma of Curcuma longa, is a widely studied phytochemical with a variety of biological activities. The ongoing research and clinical trials have proved that this natural phenolic compound has great and diverse pharmacological potencies. Beside its effective antioxidant, antiinflammatory, and antimicrobial/antiviral properties, curcumin is also considered as a cancer chemopreventive agent. While the antioxidant activity of curcumin is well documented, its interaction with DNA and RNA is not fully investigated. This study was designed to examine the interactions of curcumin with calf thymus DNA and yeast RNA in aqueous solution at physiological conditions, using constant DNA and RNA concentration (6.25?mM) and various curcumin/polynucleotide (phosphate...
Cytophotometric measurements of DNA content after Feulgen's reaction and autoradiography after incubation with "3H-thymidine were performed to investigate the early stages of development of yellow lupin root nodule. The first response of the host plant to the contact with Rhizobium lupini was the enhancement of nuclear DNA endoreplication in root cortex cells. A conversion of the initially mixoploid nodule meristem into a diploid one during root nodule formation was found. Differentiation of bacteroid-containing tissue both from diploid and polyploid cells was observed. The highest content of nuclear DNA in the uninfected root cortex, nodule cortex and in bacteroid-containing tissue was 16 C. (author).
... The thermostable Thermus aquaticus DNA polymerase, (Taq pol), in the presence of excess deoxynucleoside triphosphates (dNTPs), including deoxyadenosine ...
Adenine DNA glycosylase catalyzes the glycolytic removal of adenine from the promutagenic A {center_dot} oxoG base pair in DNA. The general features of DNA recognition by an adenine DNA glycosylase, Bacillus stearothermophilus MutY, have previously been revealed via the X-ray structure of a catalytically inactive mutant protein bound to an A:oxoG-containing DNA duplex. Although the structure revealed the substrate adenine to be, as expected, extruded from the DNA helix and inserted into an extrahelical active site pocket on the enzyme, the substrate adenine engaged in no direct contacts with active site residues. This feature was paradoxical, because other glycosylases have been observed to engage their substrates primarily through direct contacts. The lack of direct contacts in the case of MutY suggested that either MutY uses a distinctive logic for substrate ...
Synchronized cultures of the green alga Scenedesmus quadricauda were grown in the absence (untreated cultures) or in the presence (FdUrd-treated cultures) of 5-fluorodeoxyuridine, the specific inhibitor of nuclear DNA replication. The attainment of commitment points, at which the cells become committed to nuclear DNA replication, mitosis and cellular division, and the course of committed processes themselves were determined for cell cycle characterization. FdUrd-treated cultures showed nearly unaffected growth and attainment of the commitment points, while DNA replication(s), nuclear division(s) and protoplast fission(s) were blocked. Interestingly, the FdUrd-treated cells possessed a very high mitotic histone H1 kinase activity in the absence of any nuclear division(s). Compared with the ...
Several scientists have proposed that DNA repair deficiencies and the induction of a mutator phenotype are responsible for the generation of multiple mutagenic alterations in cancer cells. I propose...Full Text Available
A set of computer programs is described which constitutes a clone database management system. Maintenance of the database and the stocks of material is designed to be under the control of one person...Full Text Available
The use of europium as a local structural probe allows the various phases appearing in the NaCaPO_4-Na_3Eu(PO_4)_2 and NaSrPO_4-Na_3Eu(PO_4)_2 systems to be detected. The broadening of the europium emission lines in going from the calcium to the strontium phases illustrates the ease of displacement of the PO_4 groups. (Auth.).
1. The fluorescent fatty acid probe 11-(dansylamino)undecanoic acid (DAUDA) binds with high affinity to bovine and human serum albumin (BSA and HSA) at three sites. 2. The Kd of the primary binding...Full Text Available
Surface deposition probe techniques have been shown to be an effective means of determining the characteristics of the plasma edge region of magnetic confinement devices. Experimental results on ISX-B and EBT-S have led to a consistent picture of the edge plasma and the dominant impurity introduction mechanisms. This picture is supported by numerous other measurements and emphasizes the importance of concentrating several techniques on the complex problems of plasma fusion.
Non-destructive methods for measuring the remaining anticorrosive or antioxidative ability of coatings are investigated. For anticorrosive chromium coatings the increasing ferromagnetism caused by loss of chromium is measured by a permeability probe. Measured values on blades after operation are correlated with the progress of corrosion as determined by metallographic methods. For antioxidative aluminum coatings the diminishing layer thickness is taken as indication for the exhaustion of their protection ability. The layer thickness is measured eddy current probes, especially developed for this application.
Procedures are described for the use of synthetic oligonucleotides for Southern blot experiments and gene bank screening, and the effect of various mismatches on the efficiency of hybridization is demonstrated. The following topics are discussed: sensitivity vs. specificity, hybridization of a 12-mer to the lambda endolysin gene; hybridization of oligonucleotide probes to the E. coli lac operator; hybridization of synthetic probes to the CYC1 gene of yeast; and cloning eucaryotic genes. (HLW)
The results show that the SQUID device eddy current testing system is a suitable tool for NDE. Due to the high low-frequency sensitivity of the SQUID sensor, the SQUID device eddy current testing system permits lower examination frequencies than the conventional eddy current probe system. The SQUID system enhances fault detection in even deeper materials layers. (orig./MM).
Nondestructive inspection of carbon fibre-reinforced plastics (CRFP) using eddy current methods is based not only on the conductivity of the fibres. High frequencies (up to 10 MHz) give access to capacitive effects reflecting properties of the matrix. The paper presents a method using rotating eddy current probes to measure the anisotropic electrical properties. Potential applications of this method are the structural identification of laminates and the estimating of its degradation. (orig.)
We determined the Na/K x-ray intensity ratio in frozen sections of myocardial tissues obtained at autopsy from patients who died from various causes, using electron probe analysis. We have been able to distinguish between the ischemically injured and normal cells. The method is simple, fast, and dependable even when the duration of ischemia is only 30 minutes.
In in-service inspection of steam-generator tubes, the need for reproducibility of the measurement results and for comparison of these results over two successive inspections has led EDF to specify the main characteristics of the eddy-current probes and to design equipment to measure them. This equipment is presented here and the electrical and magnetic results are correlated with those obtained by testing tubes with reference defects.
In in-service inspection of steam-generator tubes, the need for reproducibility of the measurement results and for comparison of these results over two successive inspections has led EDF to specify the main characteristics of the Eddy-Current probes and to design equipment to measure them. This equipment is presented here and the electrical and magnetic results are correlated with those obtained by testing tubes with reference defects. 5 refs.
An eddy current probe of the level of molten metal is considered for a sectional machine of continuous casting of steel. The advantages of electromagnetic probes of the level over radiometric ones lie in their insensitivity to the layer of dross and high speed. The results and directions of further studies are given.
Geometrically anisotropic eddy current probes are a type of separate function probes especially fit to the detection of defects showing a preferential direction. This kind of flaw induces a coupling between the transmitter and the receiver by guiding eddy currents from the one to the other. On the other hand, this coupling will be almost non-existent in the presence of defects or spurious effects not displaying this geometrical particularity. Basic studies on an elementary two-coil set-up allow the acknowledgment of the intrinsic qualities of such probes: good signal-to-noise ratio, influence field practically constant on the whole defect length, ability to detect bridged defects, insensitivity to lift off. These results can be improved by achieving multicoils probes adapted to different kinds of problems. An application to continuous casting slabs testing yields very interesting results in the ...
Scanning electrochemical microscopy (SECM) was applied to evaluate the heterogeneity of a passive film formed on a pure iron electrode in deaerated pH 8.4 borate solution. A probe current image of SECM was measured with a tip-generation/sub-strate-collection (TO/SC) mode in deaerated pH 8.4 borate solution containing 0.03 moldm{sup -3} Fe(CN){sub 6}{sup 4-} as a mediator. The difference in thickness of passive films formed on two iron plates at different potentials could be evaluated from the probe current image. The probe current image of the passivated iron surface with distinctive crystal grains was composed of the patch patterns, the shapes of which coincided completely with the shapes of the substrate crystal grains. The probe current flowed above the grain surface oriented to {l_brace}100{r_brace} plane was less than that above the grain surface oriented to {l_brace}110{r_brace} or ...
To analyze the proposed growth-inhibitory function of wild-type p53, we compared simian virus 40 (SV40) DNA replication in primary rhesus monkey kidney (PRK) cells, which express wild-type p53, and...Full Text Available
BackgroundThe Mre11/Rad50/Nbs1 (MRN) complex is a regulator of cell cycle checkpoints and DNA repair. Defects in MRN can lead to defective S-phase arrest when cells are damaged....Full Text Available
Transcription for a hybrid SV40 promoter-beta globin coding sequence recombinant initiates from both early-early (EE) and late-early (LE) SV40 start sites (EES and LES) in the absence of DNA replication....Full Text Available
The role of the HCR system in the repair of prelethal lesions induced by UV light, #gamma# radiation and alkylating agents was studied in the Bacillus subtilis SPP1 phage, its heat sensitive mutants (N3, N73 nad ts_1) and corresponding infectious DNA. The survival of phages and their transfecting DNA after treatment with UV light is substantially higher in hcr"+ cells than in hcr cells, the differences being more striking in intact phages than in their transfecting DNA's. Repair inhibitors reduce survival in hcr"+ cells: caffeine lowers the survival of UV-irradiated phage SPP1 in exponentially growing hcr"+ cells but has no effect on its survival in competent hcr"+ cells; acriflavin and ethidium bromide decrease the survival of the UV-irradiated SPP1 phage in both exponentially growing and competent hcr"+ cells to the level of survival observed in hcr cells; moreover, ethidium bromide lowers the number of infective centres ...
Nucleotide pool sanitizing enzymes Dut (dUTPase), RdgB (dITPase), and MutT (8-oxo-dGTPase) of Escherichia coli hydrolyze noncanonical DNA precursors to prevent incorporation of base...Full Text Available
We have isolated and sequenced cDNA clones encoding the human U1-70K snRNP protein, and have mapped this locus (U1AP1) to human chromosome 19. The gene produces two size classes of RNA, a major 1.7-kb...Full Text Available
Previous studies using in vitro cell culture systems have shown the role of the dynamin-related GTPase Opa1 in apoptosis prevention and mitochondrial DNA (mtDNA) maintenance. However, it remains to...Full Text Available
A recombinant M13 clone (O42) containing a 65 b.p. cDNA fragment from human fetal liver mRNA coding for glyceraldehyde-3-phosphate dehydrogenase has been identified and it has been used to isolate from...Full Text Available
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
The feasibility of performing a multiplex assay for the detection of human immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV) RNAs and hepatitis B virus (HBV) DNA is demonstrated. This...Full Text Available
The histone chaperone Asf1 and the chromatin remodeler SWI/SNF have been separately implicated in derepression of the DNA damage response (DDR) genes in yeast cells treated with genotoxins that cause...Full Text Available
This short survey focuses on the main radiation-induced base lesions that have been identified within cellular DNA. For this purpose, sensitive assays that are aimed at measuring a few modifications per 10{sup 7} normal bases were set-up. In that respect high performance liquid chromatography - tandem mass spectrometry (CLHP-MS/MS) was found to be able to single out the formation of 9 oxidized nucleosides and two modified nucleo-bases out of the 70 oxidative base lesions that have been identified in model systems. As a striking result, it was found that in the DNA of {gamma}-irradiated human monocytes, the formamide-pyrimidine derivative of guanine is produced in a higher yield than the ubiquitous 8-oxo-7,8-dihydro-guanine damage, both arising from the same radical precursor. However, relatively high doses of ionizing radiation (> 20 Gy) have to be applied in order to detect an increase in the level of the damage. This is due to the low ...
By using a low-resolution macrorestriction map as the foundation (R. Majumder et al., J. Bacteriol. 176:1105–1112, 1996), an ordered cloned DNA map of the 3.2-Mb chromosome of the hypertoxinogenic...Full Text Available
A 13.1-kb DNA fragment carrying Pseudomonas denitrificans cob genes has been sequenced. The nucleotide sequence and genetic analysis revealed that this fragment contained five different cob genes named...Full Text Available
An Escherichia coli B strain, B834 galU56, has been isolated which supports growth of bacteriophage T4 with cytosine in its DNA while restricting growth of T4 with hydroxymethylcytosine. This host is...Full Text Available
The nucleotide sequence of the 5'-terminal oligonucleotides produced by pancreatic RNase digestion of bacteriophage T3 RNA polymerase (EC 2.7.7.6) transcripts of T3 DNA has been determined. The sequence...Full Text Available
We report the 1.1-Å resolution crystal structure of a bulky rhodium complex bound to two different DNA sites, mismatched and matched in the oligonucleotide 5′-(dCGGAAATTCCCG)2-3′....Full Text Available
The detection and quantification of defined reaction products of chemical mutagens and carcinogens (and of many cancer chemotherapeutic agents) with DNA require highly sensitive analytical techniques. The exceptional capability of immunoglobulins to recognize subtle alterations of molecular structure (especially when monoclonal antibodies are used to maximize specificity), outstanding sensitivity of immunoanalysis by high-affinity antibodies, and the fact that radioactively-labelled agents are not required suggest the utility of a radioimmunoassay to recognize and quantitate alkylated DNA products. We have recently developed a set of high-affinity monoclonal antibodies (secreted by mouse x mouse as well as by rat x rat hybridomas; antibody affinity constants, 10"9 to > 10"1"0 lmol) specifically directed against several DNA alkylation products with possible relevance in relation to both mutagenesis and malignant ...
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
Oxidized DNA bases, particularly 7,8-dihydro-8-oxoguanine (8-oxoG), are endogenously generated in cells, being a cause of carcinogenic mutations and possibly interfering with gene expression. We found...Full Text Available
Two integrated hepatitis B virus (HBV) DNA molecules were cloned from two primary hepatocellular carcinomas each containing only a single integration. One integration (C3) contained a single linear...Full Text Available
...Explore - The Lab - Australian Broadcasting Corporation's Gateway to Science Radio TV Shop News Sport Local Children Science Environment more Topics help Science Home News in Science Features Explore TV & Radio Dr Karl Play Podcasts DNA - The discovery of the double helix structure of DNA completely changed biology and opened up the new field of genetics. The 'backbone of life' has given us the human genome, stem cell research, ...
The endogenous reverse transcriptase reaction of equine infectious anemia virus (EIAV) has been studied, and conditions allowing synthesis of full-length minus-strand DNA have been determined. In contrast...Full Text Available
We report that expression of a nearly full-length cDNA clone of the L-A double-stranded RNA virus causes virus loss in a wild-type strain of Saccharomyces cerevisiae. We show that in this system exclusion...Full Text Available
BackgroundInfertility affects ∼20% of couples in Europe and in 50% of cases the problem lies with the male partner. The impact of damaged DNA originating...Full Text Available
One of the first technical applications of gene technology is reported here. The DNA of E. coli cells was rearranged such that these cells produce large quantities of the enzyme ..cap alpha..-galactosidase. This enzyme is used to split the trisaccharide, raffinose, which is formed during the production of sugar from sugar beet. By this means sugar yields can be increased and the energy required for sugar crystallisation reduced.
The DNA double-strand break (DSB) damage response induced by high energy charged particles on lung fibroblast cells embedded in a 3-dimensional (3-D) collagen tissue equivalents was investigated using antibodies to the DNA damage response proteins gamma-histone 2AX (#gamma#-H2AX) and phosphorylated DNA-PKcs (p-DNA-PKcs). 3-D tissue equivalents were irradiated in positions across the linear distribution of the Bragg curve profiles of 307.7 MeV/nucleon, 556.9 MeV/nucleon, or 967.0 MeV/nucleon "5"6Fe ions at a dose of 0.30 Gy. Patterns of discrete DNA damage streaks across nuclei or saturated nuclear damage were observed, with saturated nuclear damage being more predominant as samples were positioned closer to the physical Bragg peak. Quantification of the DNA damage signal intensities at each distance for each of the examined energies revealed a biological Bragg ...
32P-Postlabeling analysis and enzyme-linked immunosorbent assay (ELISA) have been used to detect DNA adducts in peripheral blood lymphocytes from primary aluminum production plant workers who were exposed...Full Text Available
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 ...
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 ...
We have identified a plasmid-like element within mitochondria of Neurospora crassa strain stp-B1. It is derived from the EcoRI-4 and EcoRI-6 regions of the mitochondrial DNA, and an additional 124 bp...Full Text Available
HMGA2 is a DNA minor-groove binding protein. We previously demonstrated that HMGA2 binds to AT-rich DNA with very high binding affinity where the binding of HMGA2 to poly(dA-dT)2 is enthalpy-driven...Full Text Available
The DNA-binding properties of Ru(bpy)2(eilatin)2+ have been investigated to determine if the sterically expansive eilatin ligand confers specificity for destabilized single-base...Full Text Available
Alkylation of DNA by 7r,8t-dihydroxy,9t,10t-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (anti-BPDE) forms mainly trans adducts (with respect to the C-9/10...Full Text Available
The nature of binding of Ru(phen) 2+ (I), Ru(bipy) 2+ (II), Ru(terpy) 2+ (III) (phen = 1,10-phenanthroline, bipy 3 = 2,2'-bipyridyl, 3 terpy = 2,2'2," - 2 terpyridyl) to DNA, poly[d(G-C)] and poly[d(A-T)]...Full Text Available
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
Yeast artificial chromosome (YAC) libraries have been difficult to construct with average insert sizes >400 kilobase pairs when DNA is size-fractionated in low-melting-point agarose. By using yeast chromosomes in mock cloning experiments, the authors found that polyamines should be present whenever agarose containing high molecular weight DNA is melted to protect DNA from degradation. By incorporating polyamines during the cloning procedure, they constructed YAC libraries from mouse and human DNA with average insert sizes of 700 and 620 kilobase pairs, respectively. Several genome equivalents of these YAC libraries were replicated onto the surface of many duplicate agar plates using a 40,000 multipin transfer device. High-density filter replicas were screened by hybridization, and 70 mouse YAC clones from 31 loci and 132 human YAC clones from 49 loci were isolated.
Fetal nucleated cells within maternal blood represent a potential source of fetal genes obtainable by venipuncture. The authors used monoclonal antibody against the transferrin receptor (TIR) to identify nucleated erythrocytes in the peripheral blood of pregnant women. Candidate fetal cells from 19 pregnancies were isolated by flow sorting at 12 1/2-17 weeks gestation. The DNA in these cells was amplified for a 222-base-pair (bp) sequence present on the short arm of the Y chromosome as proof that the cells were derived from the fetus. The amplified DNA was compared with standardized DNA concentrations. In the case of the female fetus, DNA prepared from samples at 32 weeks of gestation and cord blood at delivery also showed the presence of the Y chromosomal sequence, suggesting Y sequence mosaicism or translocation. In 10/12 cases where the 222-bp band was absent, the fetuses were female. Thus, they were ...
A new eddy current array probe has been implemented as a straightforward and promising technique for detection of outer diameter (OD) surface-breaking cracks on the extrados of feeder bends. The design is based on previous work performed at AECL, which had demonstrated that eddy current probes with laterally displaced transmit-receive coils can overcome some of the limitations of inspecting ferritic steel components for surface-breaking cracks. The Feeder Integrity Joint Program-CANDU Owners Group Inc. (FIJP-COG) Non-Destructive Evaluation (NDE) Team members commissioned AECL to work in collaboration with the probe manufacturer ZETEC, to develop a field usable eddy current array probe. The objective was to acquire a technique with the following capabilities: fast scanning non-contact inspection technique for surface breaking discontinuities; full inspection of the bend extrados OD surface in a single ...
A new eddy current array probe has been implemented as a straightforward and promising technique for detection of outer diameter (OD) surface-breaking cracks on the extrados of feeder bends. The design is based on previous work performed at AECL, which had demonstrated that eddy current probes with laterally displaced transmit-receive coils can overcome some of the limitations of inspecting ferritic steel components for surface-breaking cracks. The Feeder Integrity Joint Program-CANDU Owners Group Inc. (FIJP-COG) Non-Destructive Evaluation (NDE) Team members commissioned AECL to work in collaboration with the probe manufacturer ZETEC, to develop a field usable eddy current array probe. The objective was to acquire a technique with the following capabilities: fast scanning non-contact inspection technique for surface breaking discontinuities; full inspection of the bend extrados OD surface in a single ...
Customized reference tubes reduced sensitivity discrepancies able to be observed from one probe to the other, due to the gain setting adjustment required for a pre-definite level in amplitude response of the artificial notch. The use of a reference circuit in place of a reference part, makes characterization of the probe matched to its generator more accurate: - the material dependence is cancelled during the compensation process, - the reference signal can be adjusted more accurately in amplitude and phase response, - the manufacturing cost is reduced compared to the one necessary for machining the reference part, - the amplitude and phase response of the reference circuit can be simply modelled by using the transformer relations, such as one can appreciate the variations of the probe definition parameters and its connexion to the generator, and makes them optimal for use. The method proposed by ALSTOM for the ...
This work reports the development of an automatic methodology based on the use of 1-anilinonaphthalene-8-sulfonate (ANS) as an interfacial fluorescent probe for detecting the hydrophobic environment shift around the probe, caused by the hydrolytic action of PLA2 on the liposomes. The implementation of this reaction in a sequential injection analysis (SIA) system along with the use of the mixing chambers permitted the evaluation of PLA2 activity and assessment of the inhibitory effect of the non-steroidal anti-inflammatory drugs (NSAIDs) on PLA2 activity. Several studies were performed with the aim of establishing the appropriate flow system configuration: the liposome substrate; PLA2 and ANS optimum concentrations and incubation times before and after the enzyme addition. Based on these st...
We analyze the potential of the e{sup +}e{sup -} linear colliders, operating in the e{gamma} and {gamma}{gamma} modes, to probe anomalous quartic vector-boson interactions through the multiple production of W's and Z's. We examine all SU(2){sub L}(circle times)U(1){sub Y} chiral operators of order p{sup 4} that lead to new four-gauge-boson interactions but do not alter trilinear vertices. We show that the e{gamma} and {gamma}{gamma} modes are able not only to establish the existence of a strongly interacting symmetry breaking sector but also to probe for anomalous quartic couplings of the order of 10{sup -2} at 90% C.L. Moreover, the information gathered in the e{gamma} mode can be used to reduce the ambiguities of the e{sup +}e{sup -} mode.
We analyze the potential of the e"+e"- linear colliders, operating in the e#gamma# and #gamma##gamma# modes, to probe anomalous quartic vector-boson interactions through the multiple production of W's and Z's. We examine all SU(2)_L(circle times)U(1)_Y chiral operators of order p"4 that lead to new four-gauge-boson interactions but do not alter trilinear vertices. We show that the e#gamma# and #gamma##gamma# modes are able not only to establish the existence of a strongly interacting symmetry breaking sector but also to probe for anomalous quartic couplings of the order of 10"-"2 at 90% C.L. Moreover, the information gathered in the e#gamma# mode can be used to reduce the ambiguities of the e"+e"- mode.
A new luminescent europium probe is presented for the determination of phosphate (P) in microtiter plate format. The assay is based on the quenching of the luminescence of the europium-tetracycline (EuTc) 1:1 complex by phosphate using a reagent concentration of 20.8 {mu}mol/L. The probe is excited at 400 nm and displays a large Stokes' shift of 210 nm. The emission maximum is located at 616 nm. The system works best at neutral pH 7 and is therefore suitable for phosphate determination in biological and biochemical systems. The linear range of the calibration plot is from 5 x 10{sup -6} mol/L to 7.5 x 10{sup -4} mol/L of phosphate, and the limit of detection is 3 {mu}mol/L.
In this Brief Report, we consider dijet production at {gamma}{gamma} colliders as a probe of recently proposed, large extra-dimensional gravity models. The exchange of virtual, spin-2 graviton towers (Kaluza-Klein excitations) significantly modifies the cross section, as compared to the standard model predictions. We find that, in order to maximize the value of the effective scale that can be probed at a given center-of-mass energy, a very severe p{sub T} cut should be applied; in general, a p{sub T} equal to approximately 46% of the e{sup +}e{sup -} beam energy gives the highest reach. We find that we can probe the effective mass scale from about 2.7 TeV to 11.1 TeV, depending on the center-of-mass energy and assumptions about the model. (c) 2000 The American Physical Society.
In this Brief Report, we consider dijet production at #gamma##gamma# colliders as a probe of recently proposed, large extra-dimensional gravity models. The exchange of virtual, spin-2 graviton towers (Kaluza-Klein excitations) significantly modifies the cross section, as compared to the standard model predictions. We find that, in order to maximize the value of the effective scale that can be probed at a given center-of-mass energy, a very severe p_T cut should be applied; in general, a p_T equal to approximately 46% of the e"+e"- beam energy gives the highest reach. We find that we can probe the effective mass scale from about 2.7 TeV to 11.1 TeV, depending on the center-of-mass energy and assumptions about the model. (c) 2000 The American Physical Society
This research deals with in the study of the use of innovating magnetic sensors in eddy current non destructive inspection. The author reports an analysis survey of magnetic sensor performances. This survey enables the selection of magnetic sensor technologies used in non destructive inspection. He presents the state-of-the-art of eddy current probes exploiting the qualities of innovating magnetic sensors, and describes the methods enabling the use of these magnetic sensors in non destructive testing. Two main applications of innovating magnetic sensors are identified: the detection of very small defects by means of magneto-resistive sensors, and the detection of deep defects by means of giant magneto-impedances. Based on the use of modelling, optimization, signal processing tools, probes are manufactured for these both applications.
The ISI (In-Service Inspection) system has been developed so as to maintain the structural integrity of heat transfer tubes in the primary pressurized water cooler in the HTTR (High Temperature Engineering Test Reactor). This system consists of eddy current probes, ultra-sonic probes, insertion and extraction units, positioning unit and so on. Verification and performance tests of the developed ISI system were carried out using mock-up heat transfer tubes in the primary pressurized water cooler. The constitution of the system, R and D results of the inspection probes, and verification and performance test results of the ISI system for heat transfer tubes are described in this paper. (author)
The ISI (In-Service Inspection) system has been developed so as to maintain the structural integrity of heat transfer tubes in the primary pressurized water cooler in the HTTR (High Temperature Engineering Test Reactor). This system consists of eddy current probes, ultra-sonic probes, insertion and extraction units, positioning unit and so on. Verification and performance tests of the developed ISI system were carried out using mock-up heat transfer tubes in the primary pressurized water cooler. The constitution of the system, R and D results of the inspection probes, and verification and performance test results of the ISI system for heat transfer tubes are described in this paper. (author)
Gold and silver nanoparticles display extraordinarily large apparent refractive indices near their plasmon resonance (PR) wavelengths. These nanoparticles show good contrast in a narrow spectral band but are poorly resolved at other wavelengths in differential interference contrast (DIC) microscopy. The wavelength dependence of DIC contrast of gold/silver nanoparticles is interpreted in terms of Mie's theory and DIC working principles. We further exploit this wavelength dependence by modifying a DIC microscope to enable simultaneous imaging at two wavelengths. We demonstrate that gold/silver nanoparticles immobilized on the same glass slides through hybridization can be differentiated and imaged separately. High-contrast, video-rate images of living cells can be recorded both with and without illuminating the gold nanoparticle probes, providing definitive probe identification. Dual-wavelength DIC microscopy thus presents a new approach ...
A CANDU nuclear reactor was shut down for over one year because steam generator (SG) tubes had failed with outer diameter stress-corrosion cracking (ODSCC) in the U-bend section. Novel, single-pass eddy current transmit-receive probes, denoted as C3, were successful in detecting all significant cracks so that the cracked tubes could be plugged and the unit restarted. Significant numbers of tubes with SCC were removed from a SG in order to validate the results of the new probe. Results from metallurgical examinations were used to obtain probability-of-detection (POD) and sizing accuracy plots to quantify the performance of this new inspection technique. Though effective, the above approach of relying on tubes removed from a reactor is expensive, in terms of both economic and radiation-exposure costs. This led to a search for more affordable methods to validate inspection techniques and procedures. Methods are presented for calculating POD curves ...
A CANDU nuclear reactor was shut down for over one year because steam generator (SG) tubes had failed with outer diameter stress-corrosion cracking (ODSCC) in the U-bend section. Novel, single-pass eddy current transmit-receive probes, denoted as C3, were successful in detecting all significant cracks so that the cracked tubes could be plugged and the unit restarted. Significant numbers of tubes with SCC were removed from a SG in order to validate the results of the new probe. Results from metallurgical examinations were used to obtain probability-of-detection (POD) and sizing accuracy plots to quantify the performance of this new inspection technique. Though effective, the above approach of relying on tubes removed from a reactor is expensive, in terms of both economic and radiation-exposure costs. This led to a search for more affordable methods to validate inspection techniques and procedures. Methods are presented for calculating POD curves ...
AbstractThe structural integrity of striated muscle is determined by extra-sarcomere cytoskeleton that includes structures that connect the Z-disks and M-bands of a sarcomere to sarcomeres...Full Text Available
Many bacteria, both gram positive and gram negative, extrude in an energy-dependent manner the fluorescent pH indicator 2',7'-bis-(2-carboxyethyl)-5[and -6]-carboxyfluorescein (BCECF) (D. Molenaar,...Full Text Available
The ability to directly compare gastric ulcerated and healthy tissue would aid in the understanding of the physiological differences between these tissue types. Presently, these comparisons...Full Text Available
The homologous sequences observed for many calcium binding proteins such as parvalbumin, troponin C, the myosin light chains, and calmodulin has lead to the hypothesis that these proteins have homologous...Full Text Available
The T-loop motif is an important recurrent RNA structural building block consisting of a U-turn sub-motif and a UA trans Watson–Crick/Hoogsteen base pair. In the presence of...Full Text Available
We report the first quantitative assessment of electrosprayed droplet/ion focusing enabled by the use of a voltage-assisted air amplifier between an electrospray ionization emitter and a hybrid...Full Text Available
We describe the characteristics of the rapidly rotating molecular disk in the nucleus of the mildly active galaxy NGC4258. The morphology and kinematics of the disk are delineated by the point-like...Full Text Available
Auxin-induced pea (Pisum sativum) stem section elongation is enhanced at levels of 3 to 40 micromolar by six new classes of alkane derivatives additional to those described earlier,...Full Text Available
Failures in cortical control of fronto-striatal neural circuits may underpin impulsive and compulsive acts. In this narrative review, we explore these behaviors from the perspective of neural processes...Full Text Available
A new method is described for continuously measuring cell turgor pressure (P), hydraulic conductivity (Lp), and volumetric elastic modulus (ε) in higher plant cells, using a pressure...Full Text Available
Effects of physical environment on plasma membrane semipermeability and osmotic induction of changes in aqueous cytoplasmic volume were studied in vegetative and spore cells of a plant pathogenic fungus,...Full Text Available
The imaging analytical capabilities of laser scanning cytometer (LSC) have been used to assess morphological features considered to be typical of the senescent phenotype. The characteristic “flattening”...Full Text Available
The polarity of biological mediums controls a host of physiological processes such as digestion, signaling, transportation, metabolism, and excretion. With the recent widespread use of near-infrared...Full Text Available
Eddy current inspection methods are widely used for the nondestructive evaluation (NDE) of tubular products. Specifically, the sensors and instrumentation are designed to detect and characterize changes in a material's electrical and magnetic properties produced by the presence of discontinuities. A recent major enhancement in eddy current inspection technology has been the development of computer interfacing for data collection, analysis, and display. This breakthrough has led to multiple-frequency testing, eddy current imaging, and automated data interpretation systems that significantly enhance both capabilities and reliability of the eddy current inspection. In addition to the clear advantages in data processing, computer interfacing also permits the design and creation of unique sensors that further enhance eddy current inspection capabilities. Perhaps the most promising area of computer interactive probe design is multiple element sensors. ...
This report describes the results of EPRI Program S404-28, titled Experimental Modeling of Eddy Current Response'', conducted by the Westinghouse Science and Technology Center (STC). The Westinghouse STC demonstrates that its mercury modeling technique provides an unique bridge between steam generator eddy current field inspection conditions and predictions obtained using recently developed analytical models. The mercury modeling technique was used systematically to explore factors which contribute to primary water stress corrosion cracking (PWSCC) sizing inaccuracy, including probe design, coil excitation frequency, crack length, and crack morphology. Two new proposed techniques for inverting crack lengths from eddy current data are compared with the technique commonly used in field data analysis for PWSCC. The performance of uniform field eddy current probes is compared with the performance of pancake coil ...
This report describes the results of EPRI Program S404-28, titled Experimental Modeling of Eddy Current Response'', conducted by the Westinghouse Science and Technology Center (STC). The Westinghouse STC demonstrates that its mercury modeling technique provides an unique bridge between steam generator eddy current field inspection conditions and predictions obtained using recently developed analytical models. The mercury modeling technique was used systematically to explore factors which contribute to primary water stress corrosion cracking (PWSCC) sizing inaccuracy, including probe design, coil excitation frequency, crack length, and crack morphology. Two new proposed techniques for inverting crack lengths from eddy current data are compared with the technique commonly used in field data analysis for PWSCC. The performance of uniform field eddy current probes is compared with the performance of pancake coil ...
The localization of gold in the synovial membrane of rheumatoid arthritis patients treated with sodium aurothiomalate was examined and quantitative analysis of epon-embedded sections was carried out...Full Text Available
Despite prolonged scientific efforts to unravel the hydration structures of ions in water, many open questions remain, in particular concerning the existences and structures of ion clusters in 1∶1...Full Text Available
The main subject of this thesis is the study of the hardronic final state in deep-inelastic electron-proton scattering. Theexperiment described in this thesis is performed with such a high resolution that the substructure of the proton is probed by the electron beam with a resolution of less than 10{sup -15} cm. (orig./HSI).
The inspection of PFR evaporator tubing, superheater and reheater tubing, and tube plate examinations, are described. Ferritic steel U tubes in the evaporator have been examined by an eddy current system operating at 400 kHz using flexible rotating probes. Surface defects as shallow as 0.1 mm can be detected and sized in the range 0.1 to 0.5 mm deep. An ultrasonic method is under development for wall thickness. Special test coil probes have been developed for examination of the type 316 stainless steel superheater and reheater tubing. Crack-life defects in the bore are detectable at approximately 10% wall thickness and 20% on the outside diameter. Tube plate examinations from the tube holes, have been conducted using eddy current probes which identify surface breaking cracks in the holes. For detecting curtain cracks between tube plate holes, ultrasonic compression wave probes have been used. (U.K.).
Aug 25, 2004 ... Current Developments in Future Planetary Probe Sensors: .... Data: Outgoing and incoming EIA RS-422 interface for serial data transmission (for electrical purposes only, .... Milos, et. al, Journal of Spacecraft & Rockets 1999 ...
ObjectivesThe overall aim of this work is to use an in-situ FTIR probe to investigate selected heterogeneous catalysts in industrially relevant organic reactions. This approach will be broadly applicable to the UK fine chemical manufacturing base.~%~~%~The project has the following specific objectives:~%~~%~- To demonstrate and develop the use of an in-situ FTIR probe in a batch reactor at elevated temperatures (eg greater than 100 deg C) to monitor reactant usage and product formation.~%~~%~- To validat [continued...]DescriptionThis proposal concerns the in-situ study of catalytic processes and reaction kinetics. The catalysts concerned are microporous materials, such as, zeolites, containing pores and cavities of molecular dimensions. These catalysts constitute crystal reactors on a nanometer scale that are selective on a size and shape basis for organic molecules used in this ...
In this paper the authors show the means putting in place for examination of steam generators tubes. These means (eddy current probes, ultrasonic testing) associated with a knowledge on degradation phenomena allow mapping controlled tubes and limiting undesirable obturations.
Optical coherence microscopy (OCM) is a promising technique or high resolution cellular imaging in human tissues. An OCM system for high-speed en face cellular resolution imaging...Full Text Available
Quantitative probing of heterogeneous regions in muscle is feasible with phosphorus-31 magnetic resonance spectroscopy because of the differentiation of metabolic patterns of glycolytic and oxidative...Full Text Available
The binding of amphiphilic molecules to lipid bilayers is followed by 19F NMR using chemical shift and line shape differences between the solution and membrane-tethered states of...Full Text Available
If we decompose C1 using SVD the parameter covariance matrix can be inverted to ... However, we can analyze the parameter covariance matrix directly by ...
Detection of fatigue damage cracks in aircraft skin structures is an ongoing NDT challenge. Cracks growing rate is difficult to predict, and for this reason, it is important to detect them as soon as possible to avoid a catastrophic break. Eddy current methods are known and well-accepted techniques which are relatively simple to carry out. However, with conventional inspection of airframe structures using standard driver-pickup eddy current probes, it is difficult to detect small fatigue cracks initiated in the second layer of the skin and in the shadow region at fastener holes. Such difficulty is related to fasteners length and their spacing which reduces test sensitivity. This report explains the development of improved eddy current probe/technique to detect and visualize fatigue cracks in fasteners. In this study, an eddy current array probe was evaluated and new coil design was suggested. Several tests were conducted on ...
13C metabolic flux analysis (MFA) is the most comprehensive means of characterizing cellular metabolic states. Uniquely labeled isotopic tracers enable more focused analyses to probe...Full Text Available
Although ultrasonic testing inspection technology and tools have improved significantly, there is still a need for more reliable detection, monitoring, and accurate sizing of crack-like and planar defects, complex corrosion damage, and detection of secondary features within deformed pipe. Ultrasonic two dimensional (2D) matrix phased array technology offers some unique advantages that make the technology promising for improving detection and sizing of pipeline flaws resulting from welding or from in-service damage. Ultrasonic modeling and simulation has been conducted to evaluate the detection and sizing capabilities of 2D matrix arrays for various pipeline inspection concepts. Simulations have been performed using both flexible and rigid array probes. Inspection concepts using rigid probes were evaluated for inspections from both the outside and inside pipe surfaces, while flexible probes were evaluated primarily for ...
The dissociation of methylal is investigated using mass spectrometry, combined with a pyrolytic radical source and femtosecond pump probe experiments. Based on preliminary results two reaction paths of methylal dissociation are proposed and discussed. (author) 4 fig., 3 refs.
Various physico-chemical tissue optical clearing (TOC) methods have been suggested to maximize photon density in tissue. In order to enhance photon density, a compression-controlled low-level...Full Text Available
Improved NMR detection of mass limited samples can be obtained by taking advantage of the mass sensitivity of microcoil NMR, while throughput issues can be addressed using multiple, parallel...Full Text Available
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
Heat transfer tubes of a primary pressurized water cooled (PPWC) in the high temperature engineering test reactor (HTTR) form the reactor pressure boundary of the primary coolant, therefore are important from the viewpoint of safety. To establish inspection techniques for the heat transfer tubes of the PPWC, an automatic inspection system was developed. The system employs a bobbin coil probe, a rotating probe for eddy current testing (ECT) and a rotating probe for ultrasonic testing (UT). Nondestructive test of a half of the heat transfer tubes of the PPWC was carried out by the automatic inspection system during reactor shutdown period of the HTTR (about 55% in the maximum reactor power in this paper). The nondestructive test results showed that the maximum signal-to-noise ratio was 1.8 in ECT. Pattern and phase of Lissajous wave, which were obtained for the heat transfer tube of the PPWC, were different from those ...
Experimental data are reported on the micro-irregularities of resistivity in longitudinal and transverse specimens of germanium 0.5 to 0.8 mm in thickness and 25 to 40 mm in width. These data are correlated with the conditions of growth of the crystals. (16 refs).
BackgroundThe pH is an important parameter controlling many metabolic and signalling pathways in living cells. Recombinant fluorescent pH indicators (pHluorins) have come into vogue...Full Text Available
Water-maze testing has been used to assess prenatal cocaine (PCOC)- induced deficits in behavioral studies of spatial navigation and memory abilities. Effects of PCOC in acquisition or in probe...Full Text Available
Mutation C1494T in mitochondrial 12S rRNA gene was recently reported in two large Chinese families with aminoglycoside-induced and nonsyndromic hearing loss (AINHL) and was claimed to be pathogenic. This mutation, however, was first reported in a sample from central China in our previous study that was aimed to reconstruct East Asian mtDNA phylogeny. All these three mtDNAs formed a subclade defined by mutation C1494T in mtDNA haplogroup A. It thus seems that mutation C1494T is a haplogroup A-associated mutation and this matrilineal background may contribute a high risk for the penetrance of mutation C1494T in Chinese with AINHL. To test this hypothesis, we first genotyped mutation C1494T in 553 unrelated individuals from three regional Chinese populations and performed an extensive search ...
An enantiocontrolled route to aziridinomitosenes had been developed from l-serine methyl ester hydrochloride. The tetracyclic target ring system was assembled by an internal azomethine ylide cycloaddition reaction based on silver ion-assisted intramolecular oxazole alkylation and cyanide-induced ylide generation via a labile oxazoline intermediate (62 to 66). Other key steps include reductive detritylation of 26, methylation of the N-H aziridine 56, oxidation of the sensitive cyclohexenedione 68 to quinone 70, and carbamoylation using Fmoc-NCO. Although the aziridinomitosene tetracycle is sensitive, a range of protecting group manipulations and redox chemistry can be performed if suitable precautions are taken. A study of DNA alkylation by the first C-6,C-7-unsubstituted aziridinomitosene 11a has been carried out, and evidence for DNA cross-link formation involving nucleophilic addition to the quinone subunit is described. PMID:14677970
Retroelements constitute a large part of the human genome. These sequences are mostly silenced in normal cells, but genome-wide DNA hypomethylation in cancers might lead to their re-expression. Whether this re-expression really occurs in human cancers is largely unkown. We therefore investigated expression and DNA methylation of several classes of retroelements in human prostate cancer tissues and cell lines by quantitative reverse transcription-polymerase chain reaction and pyrosequencing, respectively. The most striking finding was strong and generalized increased expression of the HERV-K_22q11.23 provirus in cancers, including de novo expression of a spliced accessory Np9 transcript in some tumors. In parallel, DNA methylation in the long terminal repeat (LTR) decreased. Conversely, HER...
Abstract Ultrastructural studies have shown that the formation of pigment glands in Gossypium hirsutum L. leaves is a lysigenous process, originating from a cluster of cells in the ground meristem. Various techniques were used here to investigate whether programmed cell death (PCD) plays a critical role in this developmental process. Nuclei of internal cells in the pigment gland-forming tissue were TUNEL-positive and DAPI-negative, suggesting that DNA cleavage is an early event and complete DNA degradation is a late event. Smeared bands and a lack of laddering after gel electrophoresis indicate that DNA cleavage is random. Ultrastructurally, secretory cells in the pigment glands become distorted, nuclei are densely stained, and chromosomes become condensed until completely degraded at late...
We describe the cloning and characterization of a mutated thermostable DNA polymerase from Thermus aquaticus (Taq) that exhibits an increased reverse transcriptase activity and is therefore designated for one-step PCR pathogen detection using established real-time detection methods. We demonstrate that this Taq polymerase mutant (Taq M1) has similar PCR sensitivity and nuclease activity as the respective Taq wild-type DNA polymerase. In addition, and in marked contrast to the wild-type, Taq M1 exhibits a significantly increased reverse transcriptase activity especially at high temperatures (>60degreeC). RNA generally hosts highly stable secondary structure motifs, such as hairpins and G-quadruplexes, which complicate, or in the worst case obviate, reverse transcription (RT). Thus, RT at hi...
Fasciolosis caused by Fasciola spp. (Platyhelminthes: Trematoda: Digenea) is considered as the most important helminth infection of ruminants in tropical countries, causing considerable socioeconomic problems. In the endemic regions of the North of Iran, Fasciola hepatica and Fasciola gigantica have been previously characterized on the basis of morphometric differences, but the use of molecular markers is necessary to distinguish exactly between species and intermediate forms. Samples from buffaloes and goats from different localities of northern Iran were identified morphologically and then genetically characterized by sequences of the first (ITS-1) and second (ITS-2) Internal Transcribed Spacers (ITS) of nuclear ribosomal DNA (rDNA). Comparison of the ITS of the northern Iranian samples ...
We investigate quantum mechanical electron transport along the long axis of the DNA molecule using an effective tight-binding model. The overall contour plot of transmission, the current-voltage characteristics, and the differential conductance are examined for the variation of backbone onsite energy, the energy-dependent hopping strength, and the contact coupling between the leads and the DNA molecule. It is shown that as backbone asymmetry increases, the merging and collapse of the two mini-bands take place and an extra resonance peak in the transmission appears. In addition, we present the modulation of voltage threshold in the current-voltage curves and a double-peak structure in the differential conductance due to the disappearance of the merged mini-band. Finally, in the Coulomb bloc...
The antioxidant properties of magnolol and honokiol were evaluated in the experimental systems of reducing ONOO^- and ^1O"2, bleaching @b-carotene in linoleic acid (LH) emulsion, and trapping 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) cationic radical (ABTS^+?) and 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH), and then were applied to inhibit the oxidation of DNA induced by Cu^2^+/glutathione (GSH) and 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH). Magnolol and honokiol were active to reduce ONOO^- and ^1O"2. Honokiol showed a little higher activity to protect LH and to inhibit Cu^2^+/GSH-induced oxidation of DNA than magnolol. In addition, honokiol exhibited higher activities to trap ABTS^+? and DPPH than magnolol. In particular, honokiol trapped 2.5 radicals while magnolol o...
A clone encoding a human D{sub 2} dopamine receptor was isolated from a pituitary cDNA library and sequenced. The deduced protein sequence is 96% identical with that of the cloned rat receptor with one major difference: the human receptor contains an additional 29 amino acids in its putative third cytoplasmic loop. Southern blotting demonstrated the presence of only one human D{sub 2} receptor gene. Two overlapping phage containing the gene were isolated and characterized. DNA sequence analysis of these clones showed that the coding sequence is interrupted by six introns and that the additional amino acids present in the human pituitary receptor are encoded by a single exon of 87 base pairs. The involvement of this sequence in alternative splicing and its biological significance are discussed.
Background: A possible association between the polymorphic CAG repeat in the DNA polymerase gamma (POLG) gene and the risk of testicular germ-cell tumours (TGCT) was investigated in this study. The hypothesis was prompted by an earlier preliminary study proposing an association of the absence of the common 10-CAG-long POLG allele with testicular cancer as well as previously reported in some European populations' association with male subfertility, which is a condition carrying an increased risk of TGCT. Patients and methods: The number of CAG repeats in both POLG alleles was established in 243 patients with TGCT and in 869 controls by the analysis of the genomic DNA fragment. Results: A significantly higher proportion of men homozygous allele of other than the common 10 CAG repeats was fou...
We present two applications of CR-39 to estimate biological effects of heavy ion irradiation. The accurate measurement of fluence of ions using CR-39 is indispensable to calculate the action cross sections for biological effects. Ions with 6 MeV/n at the Medium Energy Beam Course, HIMAC (NIRS) were extracted to the air, and degraded with the air. DNA and living cells were irradiated by ions with various specific energies at several air columns along with the Bragg curve. DNA strand breaks and cell killing were measured and the results were converted to the action cross sections using the fluence measured with CR-39 at the irradiation positions. Another example of the application of CR-39 is to identify whether the ions with a specific energy pass through the cell or stop within the cell. (author)
Methylation of the DNA bases in the Watson-Crick GC and AT base pairs by the methyldiazonium ion was investigated employing density functional and second order Moller-Plesset (MP2) perturbation theories. Methylation at the N3, N7 and O6 sites of guanine, N1, N3 and N7 sites of adenine, O2 and N3 sites of cytosine and the O2 and O4 sites of thymine were considered. The computed reactivities for methylation follow the order N7(guanine)>N3(adenine)>O6(guanine) which is in agreement with experiment. The base pairing in DNA is found to play a significant role with regard to reactivities of the different sites.
We calculate quark-antiquark potentials using the relationship between the expectation value of the Wilson loop and the action of a probe string in the string dual. We review and categorize the possible forms of the dependence of the energy on the separation between the quarks. In particular, we examine the possibility of there being a minimum separation for probe strings which do not penetrate close to the origin of the bulk space, and derive a condition which determines whether this is the case. We then apply these considerations to the flavoured resolved deformed conifold background of Gaillard et al. We suggest that the unusual behaviour we observe in this solution is likely to be related to the IR singularity which is not present in the unflavoured case.
The use of europium as a local structural probe allows the various phases appearing in the NaCaPO/sub 4/-Na/sub 3/Eu(PO/sub 4/)/sub 2/ and NaSrPO/sub 4/-Na/sub 3/Eu(PO/sub 4/)/sub 2/ systems to be detected. The broadening of the europium emission lines in going from the calcium to the strontium phases illustrates the ease of displacement of the PO/sub 4/ groups.
Results are presented applying an enteral nutrition by means of probes with Cosilat in complex therapy of patients with radiotherapeutic late effects of the colon. This nutrition was prescribed with the intention to gave the colon a functional recovery and to guarantee optimal conditions for repair of radiation injuries. Good results were seen in 27 of 28 patients (96.7%), the situation of one patient (3.3%) continued without any change. Cosilat has a high nutritive value, good organoleptic qualities and is well digested without any complications. (author).
Results are given for deep-level petroleum and gas drilling probes in various coastal zones of Southern Sakhalin covering the Poronai depression and the Aniv Bay trough. The exploratory probes indicate that the Middle Miocence deposits of the Boundary trough contain commercial-size deposits of petroleum, that commercial-size gas deposits are in the upper Myocene deposits of the Aliv Bay trough, and that the upper Cretaceous and Cenozoic deposits of the eastern section of Southern Sakhalin lack folded deformations. 7 references, 2 figures.
Surface probe and laser-induced fluorescence measurements in ISX-B and EBT-S have made significant contributions to the understanding of plasma edge characteristics and plasma-surface interactions in these devices. Where comparison is possible, these techniques have led to results which are consistent with plasma diagnostics. Charge-exchange neutral sputtering and self-ion sputtering have been identified as the dominent heavy impurity release mechanisms in ISX-B and EBT-S, respectively.
Secondary-electron emission (SEE) spectra have been obtained with the Scanning Probe Energy Loss Spectrometer at a tip-sample distance of only 50 nm. Such short working distances are required for the best theoretical spatial resolution (<10 nm). The SEE spectra of graphite, obtained as a function of tip bias voltage, are shown to correspond to unoccupied states in the electronic band structure. The SEE spectra of thin gold films demonstrate the capability of identifying (carbonaceous) surface contamination with this technique.
Individual laser cooled atoms are delivered on demand from a single atom magneto-optic trap to a high-finesse optical cavity using an atom conveyor. Strong coupling of the atom with the cavity field allows simultaneous cooling and detection of individual atoms for time scales exceeding 15 s. The single atom scatter rate is studied as a function of probe-cavity detuning and probe Rabi frequency, and the experimental results are in good agreement with theoretical predictions. We demonstrate the ability to manipulate the position of a single atom relative to the cavity mode with excellent control and reproducibility.
This paper describes detection of electro-discharged machine (EDM) defects in magnetic steam generator (SG) tubes of Monju fast breeder reactor (FBR). The EDM defects are located under support plate (SP), on the outer tube surface and they are detected by a remote field eddy current probe. Using the experimental measurements and a multi frequency algorithm, the defect signal can be extracted from the SP signal. The parameters of the multi-frequency algorithm were calculated by comparing SP measurements with two-dimensional finite element simulations (FEM). (author)
A proposal is made to test Newton's inverse-square law using the perihelion shift of test masses (planets) in free fall within a spacecraft located at the Earth-Sun L2 point. Such an Artificial Planetary System In Space (APSIS) will operate in a drag-free environment with controlled experimental conditions and minimal interference from terrestrial sources of contamination. We demonstrate that such a space experiment can probe the presence of a "hidden" fifth dimension on the scale of a micron, if the perihelion shift of a "planet" can be measured to sub-arc-second accuracy. Some suggestions for spacecraft design are made.
In one of the first uses of high Tc superconductors in an end-use product, Lake Shore Cryotronics and a Swiss lab have developed a liquid nitrogen level sensor using a high Tc thin film. The probe is manufactured using a seamless stainless steel tube with an yttrium-based zirconium oxide flame-sprayed on the tube. A plasma-sprayed superconductor compound is deposited on top of that. The probe is coated with a sealant that protects the superconducting film from the atmosphere. This manufacturing method has yielded an extremely durable product. Unaffected by ice formation and related mechanical problems, the superconducting level sensor can be kept in the dewar for long periods of time.
\\lhcb is one of the four major experiments at the Large Hadron Collider (\\lhc) at CERN. It is custom built to look for CP violation and New Physics in rare decays of heavy flavour hadrons, like the B and D systems. Rare decays that occur via loop diagrams provide a way to probe New Physics at energy scales much higher than can be probed by direct production in experiments. In this article, the \\lhcb prospects for such measurements with exclusive decays of the type $B \\to \\Xs \\gamma$ and $B \\to X_{s} l^{+} l^{-}$ are presented.
LHC$b$ is one the four major experiments at the Large Hadron Collider (LHC) at CERN. It is custom built to look for CP violation and New Physics in rare decays of heavy flavor hadrons, like the B and D systems. Rare decays that occur via loop diagrams provide a way to probe New Physics at energy scales much higher than can be probed by direct production in experiment. In this article, the LHC$b$ prospects for such measurement with exclusive decays of the type $B \\to X_s\\gamma$ and $B\\to X_sl^+l^-$ are presented.
Eddy current testing (ECT) method is widely used as the non-destructive evaluation (NDE) of the various kinds of material degradation occurring in nuclear power plants (NPPs) components including steam generator (SG) tubes. Detection of any defects or flaws in the steam generator tubes in the early stage is very important in maintenance of NPP for its primary role as the pressure boundary with thin wall thickness. Although the ECT technique provides lots of information for a SG management, it has a generic problem in its reliability due to a low ability in detect of small defects and some difficulty in a signal analysis. For the improvement of these shortcomings in conventional ECT, profile-MRPC (motorized rotating pancake coil) ECT technology was developed in KAERI. The key of this new technology is the development of new eddy current probes, designated as a diagnostic probe (D-probe). The D-probe is ...
Eddy current testing (ECT) method is widely used as the non-destructive evaluation (NDE) of the various kinds of material degradation occurring in nuclear power plants (NPPs) components including steam generator (SG) tubes. Detection of any defects or flaws in the steam generator tubes in the early stage is very important in maintenance of NPP for its primary role as the pressure boundary with thin wall thickness. Although the ECT technique provides lots of information for a SG management, it has a generic problem in its reliability due to a low ability in detect of small defects and some difficulty in a signal analysis. For the improvement of these shortcomings in conventional ECT, profile-MRPC (motorized rotating pancake coil) ECT technology was developed in KAERI. The key of this new technology is the development of new eddy current probes, designated as a diagnostic probe (D-probe). The D-probe is ...
Moisture profiles from neutron probe data provide valuable information in site characterization and to supplement ground water monitoring efforts. The neutron probe precision error (reproducibility) is found to be about 0.2 vol% under in situ field conditions where the slope in moisture content with depth is varying slowly. This error is about 2 times larger near moisture spikes (e.g., at the vapor phase notch), due to the sensitivity of the probe response to vertical position errors on the order of 0.5 inches. Calibrations were performed to correct the downhole probe response to the volumetric moisture content determined on core samples. Calibration is sensitive to borehole diameter and casing type, requiring 3 separate calibration relations for the boreholes surveyed here. Power law fits were used for calibration in this study to assure moisture content results greater than zero. Findings in the ...
The WEGA stellarator is used to confine low temperature, overdense (densities exceeding the cut-off density of the heating wave) plasmas by magnetic fields in the range of B=50-500 mT. Microwave heating systems are used to ignite gas discharges using hydrogen, helium, neon or argon as working gases. The produced plasmas have been analyzed using Langmuir and emissive probes, a single-channel interferometer and ultra-high resolution Doppler spectroscopy. For a typical argon discharge in the low field operation, B=56 mT, the maximum electron density is n{sub e}{proportional_to}10{sup 18} m{sup -3} with temperatures in the range of T=4-12 eV. The plasma parameters are determined by using Langmuir probes and are cross-checked with interferometry. It is demonstrated within this work that the joint use of emissive probes and ultra-high resolution Doppler spectroscopy allows a precise measurement of the radial electric field. The ...
BackgroundPuccinia striiformis f. sp. tritici is a fungal pathogen causing stripe rust, one of the most important wheat diseases worldwide. The...Full Text Available
Global acetylation of histone H4 is a mark of gene transcriptional activation. The c-Myc transcription factor binds to specific DNA sites in cellular chromatin and induces the acetylation of...Full Text Available
Recently, a novel DNA virus designated SEN virus (SEN-V), which is thought to be related to posttransfusion hepatitis, was discovered. The aim of the present study was to clarify the relationship between...Full Text Available
Significant emphasis has recently been placed on the characterization of the human cancer genome. This effort has been assisted by the development of new DNA sequencing technologies that allow...Full Text Available
FRAXA is one of a number of fragile sites in human chromosomes that are induced by agents like fluorodeoxyuridine (FdU) that affect intracellular thymidylate levels. FRAXA coincides with a >200...Full Text Available
The LW blood group antigens reside on a 42-kDa erythrocyte membrane glycoprotein that was purified by immunoaffinity and partially sequenced. From this information, a specific PCR-amplified DNA fragment...Full Text Available
Activation-induced deaminase (AID) initiates somatic hypermutation, gene conversion and class switch recombination by deaminating variable and switch region DNA cytidines to uridines. AID is predominantly...Full Text Available
The accepted model of retroviral reverse transcription includes a circular DNA intermediate which requires strand displacement synthesis for linearization and creation of an integration-competent, long...Full Text Available
Using the complete genome of Plasmodium falciparum 3D7 which has 14 chromosomes as an example, we have examined the distribution functions for the amount of C or G and A or T consecutively...Full Text Available
BackgroundGenomes store information for building and maintaining organisms. Complete sequencing of many genomes provides the opportunity to study and compare global information properties...Full Text Available
Nuclear hormone receptors comprise a characteristic family of transcription factors found in vertebrates, insects and nematodes. Here we show by cDNA and gene cloning that a Cnidarian, Tripedalia...Full Text Available
We have used restriction endonuclease analysis (REA) of genomic DNA to classify porcine Pasteurella multocida isolates with similar capsular and somatic serotypes, and to monitor the distribution of...Full Text Available
BackgroundEukaryotic DNA replication is regulated at the level of large chromosomal domains (0.5–5 megabases in mammals) within which replicons are activated relatively synchronously....Full Text Available
Reverse genetics technology so far established for measles virus (MeV) is based on the Edmonston strain, which was isolated several decades ago, has been passaged in nonlymphoid cell lines, and is no...Full Text Available
Gene expression is a unique way of characterizing how cells and organisms adapt to changes in the external environment. The measurements of gene expression levels upon exposure to a chemical can be...Full Text Available
A rapid, sensitive, and simple method was developed to detect the sapstain fungi Ophiostoma piceae and O. quercus in stained wood. By using microwave heating for DNA...Full Text Available
A pore-forming peptide is implicated in the potent cytolytic activity of pathogenic Entamoeba histolytica. Using NH2-terminal sequence information of this peptide, the corresponding cDNA was isolated....Full Text Available
Mapping the restriction fragments of the Brucella melitensis 16M genome with a new restriction endonuclease, PacI, which cut the DNA into only eight fragments, indicated that this species contains two...Full Text Available
Erythrovirus (formerly parvovirus) B19 causes a wide range of diseases in humans, including anemia due to aplastic crisis. Diagnosis of B19 infection relies on serology and the detection of viral DNA...Full Text Available
The chaperone-encoding groESL and dnaK operons constitute the CIRCE regulon of Bacillus subtilis. Both operons are under negative control of the repressor...Full Text Available
BackgroundMutations that impair mitochondrial functioning are associated with a variety of metabolic and age-related disorders. A barrier to rigorous tests of the role of mitochondrial...Full Text Available
A plasmid carrying the bacteriophage lambda lysis genes under lac control was subjected to hydroxylamine mutagenesis, and mutations eliminating the host lethality of the S gene were selected. DNA sequence...Full Text Available
The Far UpStream Element (FUSE) regulatory system promotes a peak in the concentration of c-Myc during cell cycle. First, the FBP transcriptional activator binds to the FUSE DNA element upstream...Full Text Available
Chemical reactions in living cells are under strict enzyme control and conform to a tightly regulated metabolic program. However, uncontrolled and potentially deleterious endogenous reactions occur,...Full Text Available
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
In vitro mutagenesis of functional DNA gene fragments by covalently reactive agents permits one in principle to examine the consequent alterations in DNA sequence directly. I have carried out selective mutagenesis of the tetracycline resistance gene in the plasmid pBR322 using the long wavelength UV light activated reaction of 4,5',8-trimethylpsoralen (TMP). The mutagenized DNA sequence was the EcoR1-Hind III restriction fragment in the vicinity of the Tcsup(R) promoter. Two classes of mutants were obtained. One exhibited a high level of Tc resistance (40-60 ..mu..g/ml) but still lower than the wild-type. Interestingly, these showed no sequence alterations at all in the vicinity of the TMP-reacted fragment. The other class of mutants exhibited low levels of drug resistance (< 20 ..mu..g/ml) and two of those that were sequenced were found to contain a 15-base pair insertion to the right of the original Hind III ...
BackgroundRecently, the discovery of copy number variation (CNV) led researchers to think that there are more variations of genomic DNA than initially believed. Moreover, a certain...Full Text Available
A duplication of the polypurine tract (PPT) at the center of the human immunodeficiency virus type 1 (HIV-1) genome (the cPPT) has been shown to prime a separate plus-strand initiation and to result...Full Text Available
The DNA sequences were determined for the lipoprotein lipase (LPL) gene from five unrelated Japanese patients with familial LPL deficiency. The results demonstrated that all five patients are homozygotes...Full Text Available
Forum domains are stretches of chromosomal DNA that are excised from eukaryotic chromosomes during their spontaneous non-random fragmentation. Most forum domains are 50–200 kb in length....Full Text Available
BackgroundDeveloping lepidopteran microsatellite DNA markers can be problematical, as markers often exhibit multiple banding patterns and high frequencies of non-amplifying "null"...Full Text Available
Werner Syndrome (WS) is an inherited disease characterized by premature onset of aging, increased cancer incidence, and genomic instability. The WS gene encodes a 1,432-amino acid polypeptide (WRN)...Full Text Available
AIM: To analyze the antiviral mechanism of Epigallocatechin gallate (EGCG) against hepatitis B virus (HBV) replication.METHODS: In this research, the HBV-replicating cell line HepG2.117 was used...Full Text Available
IntroductionRadiotherapy outcomes might be further improved by a greater understanding of the individual variations in normal tissue reactions that determine tolerance. Most published...Full Text Available
Experiments were performed to investigate the involvement of the cell membrane in the excision DNA repair process in Escherichia coli. Two membrane-binding drugs, procaine and phenethyl alcohol (PEA), inhibited liquid-holding recovery (LBR) in u.v.-irradiated E. coli wild-type and recA strains. In uvrB and polA strains where, after u.v.-irradiation, LHR was absent the two drugs had no effect. Both drugs markedly reduced the removal of u.v.-induced thymine dimers in the DNA of wild-type cells (H/r30). Analysis by alkaline sucrose gradients revealed that PEA inhibited the incision step in excision repair. In contrast, procaine had no effect on incision but apparently inhibited the late steps in excision repair. PEA dissociated DNA from the cell membrane, whereas procaine did not. The results suggest that the two drugs PEA and procaine inhibit LHR and the excision repair process operating on u.v.-induced damage in E. coli by ...
One of the central questions of molecular biology is the discovery of the semantics of DNA. This discovery relies in a critical way on a variety of expensive computations. In order to solve these computations, both parallel computers and special-purpose hardware play a major role.
BackgroundPolyethyleneimine (PEI), which can interact with negatively charged DNA through electrostatic interaction to form nanocomplexes, has been widely attempted to use as a gene...Full Text Available
In methicillin-resistant Staphylococcus aureus, the methicillin resistance gene mecA is localized within a large chromosomal region which is absent in the methicillin-susceptible...Full Text Available
The functional gene and three intronless pseudogenes for human triosephosphate isomerase were isolated from a recombinant DNA library and characterized in detail. The functional gene spans 3.5 kilobase...Full Text Available
We have cloned and characterized the entire DNA polymerase gene and flanking regions from Kaposi's sarcoma-associated herpesvirus (KSHV) and two closely related macaque homologs of KSHV, retroperitoneal...Full Text Available
BackgroundHealth risk from exposure of perfluorochemicals (PFCs) to wildlife and human has been a subject of great interest for understanding their molecular mechanism of toxicity....Full Text Available
BACKGROUND: DNA prenatal diagnosis for cystic fibrosis (CF) has been available for parents of affected children since late 1985. METHODS: Using anonymous questionnaires, we surveyed 395 parents of children...Full Text Available
In biotechnology, acrylamide is being used in DNA and RNA analysis using the polyacrylamide gel electrophoreses procedure. Polymerized acrylamide is degraded into acrylamide through time; it is converted into a hazardous contaminant that is carcinogenic and neurotoxic to animals and humans. Because ...
Chicken embryos were x-irradiated in ovo with 5-30 cGy (=priming dose) at the 13th-15th day of development. After 3-48 h, brain- and liver-cell suspensions were x-irradiated in vitro with (challenge) doses of 4-32 Gy. Significantly less radiation damage was observed when the radiation response was measured by scheduled DNA synthesis, nucleoid sedimentation and viscosity of alkaline cell lysates 12-36 h after the priming exposure. In vivo, pre-irradiation with 10 cGy enhanced regeneration as evidenced by the DNA content of chicken embryo brain and liver 24 h following a challenge dose of 4 Gy. From nucleoid sedimentation analyses in brain and liver cells immediately after irradiation with 16 Gy and after a 30-min repair period in the presence of aphidicolin, dideoxythymidine and 3-aminobenzamide or in the absence of these DNA repair inhibitors, it is concluded that a reduction of the initial radiation damage is the dominant ...
SummaryActivating transcription factor 2 (ATF2) is regulated by JNK/p38 in response to stress. Here, we demonstrate that the protein kinase ATM phosphorylates ATF2 on serines...Full Text Available
For identification of genes responsible for varietal differences in flowering time and leaf morphological traits, we constructed a linkage map of Brassica rapa DNA markers including...Full Text Available