Science.gov (United States)

... The mission of Tandem Thrust is to employ a joint task force headquarters and appropriate forces in a realistic training exercise. ...

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Computer simulations of two-junction, concentrator tandem solar cell performance show that IR-sensitive bottom cells are required to achieve high efficiencies. Based on this conclusion, two novel concentrator tandem designs are under investigations: (1) a mechanically stacked, four-terminal GaAs/GaInAsP (0.95 eV) tandem, and (2) a monolithic, lattice-matched, three-terminal InP/GaInAs tandem. In preliminary experiments, terrestrial concentrator efficiencies exceeding 30% have been achieved with each of the above tandem designs. Methods for improving the efficiency of each tandem type are discussed. (orig.).

UK PubMed Central (United Kingdom)

A liquid chromatography/tandem mass spectrometric (LC/MS/MS) assay was developed for the quantitative determination of salirasib (S-trans, trans-farnesylthiosalicylic acid,...Full Text Available

International Nuclear Information System (INIS)

The motivation for using a tandem electrostatic accelerator as an electron accelerator for a free electron laser (FEL) is presented. The adaptation of the HVEC EN tandem at the Weizmann Institute for this purpose, electron beam optics and nonlinear FEL computation relevant for this FEL realization are described. In the tandem configuration the terminal is held at a positive potential. The electron beam is accelerated from ground potential to the terminal in one beam tube and then decelerated down the other beam tube. The FEL wiggler and cavity are at the terminal. Due to the beam energy recovery this scheme produces a high power beam at the terminal with a small investment in electrical power. (orig.).

Science.gov (United States)

Group B Streptococcus

UK PubMed Central (United Kingdom)

In clinical situations where a diagnostic real-time PCR assay is not sensitive enough, leading to low or falsely negative results, or where detection earlier in a disease progression would benefit the...Full Text Available

International Nuclear Information System (INIS)

Borrelia burgdorferi sensu lato genospecies were differentiated by PCR-based sequencing of the borrelial flagellin gene. To evaluate the usefulness of KingFisher"T"M magnetic particle processor in PCR product purification, borrelia PCR products were purified with KingFisher"T"M magnetic particle processor prior to cycle sequencing and the quality of the sequence data received was analyzed. KingFisher was found to offer a rapid and reliable alternative for borrelial PCR product purification.

Energy Technology Data Exchange (ETDEWEB)

The various factors that affect the open-circuit voltage ([ital V][sub oc]) of a two-terminal GaInP[sub 2]/GaAs tandem cell are examined. These include a) an anomalous problem associated with the GaAs bottom cell and b) back surface passivation of the thin GaInP[sub 2] top cell. Solutions to these problems are presented and yield tandem [ital V][sub oc]s close to the practical theoretical limits.

UK PubMed Central (United Kingdom)

SummaryBlastomycosis is a serious and potentially fatal infection by the thermally dimorphic fungus Blastomyces dermatitidis. PCR assays targeting the BAD-1 virulence...Full Text Available

UK PubMed Central (United Kingdom)

Since current microbiology methods are not suitable to detect Clostridium perfringens in formalin-fixed, paraffin-embedded tissue samples, we developed a PCR assay to detect toxin-encoding...Full Text Available

UK PubMed Central (United Kingdom)

A commercial multiplex real-time PCR assay (Cepheid Xpert C. difficile assay) for the diagnosis of Clostridium difficile infection was evaluated. The sensitivity and...Full Text Available

Science.gov (United States)

... R OFFICIAL FILE COPY SYNTHESIS OF PERFLUOROALIPHATIC ETHER MONOMERS PCR, Inc. ... Synthesis of Perfluoroaliphatic Ether Monomers ...

Science.gov (United States)

Invasive Aspergillosis

Energy Technology Data Exchange (ETDEWEB)

A description is given of the principal steps involved in `USIMINAS` work roll manufacturing project for its tandem cold mill. The service performance of these rolls was found to be similar to that rolls supplied by traditional manufacturers. (author). 5 refs., 5 figs., 3 tabs.

Energy Technology Data Exchange (ETDEWEB)

This annual report describes research activities which have been performed with the JAERI tandem accelerator and the Van de Graaff accelerator from April 1, 2000 to March 31, 2001. Summary reports of 46 papers, and lists of publication, personnel and cooperative research with universities are contained. (author)

Energy Technology Data Exchange (ETDEWEB)

The electrical transport taking place in the {mu}c-Si tunnel recombination junction (TRJ) of a-Si:H/a-Si:H tandem solar cells and the role of CO{sub 2} plasma oxidation performed between microcrystalline layers is investigated in this paper with the computer code AMPS. Oxidized interfaces were modelled as simple highly defective intrinsic {mu}c-Si layers. Two different tunnel junction structures are studied in this paper: (a) (n){mu}c-Si/oxide/(p){mu}c-Si and (b) (n){mu}c-Si/(i){mu}c-Si/(p){mu}c-Si. In the last configuration the oxide interface is removed and replaced by a thin defective (i) {mu}c-Si layer. Both tunnel junctions have comparable theoretical and experimental tandem solar cell efficiencies which indirectly proves that our modelling assumption for oxidised interfaces is correct. A-Si:H/a-Si:H tandem solar cell efficiencies depend on the thickness of the intrinsic layer introduced in the tunnel junction. The ...

British Library Electronic Table of Contents (United Kingdom)

A simple, sensitive and rapid method for analysis of granisetron in human plasma, utilizing liquid chromatography tandem mass spectrometry (LC-MS/MS), has been developed and validated to satisfy FDA guidelines for bioanalytical methods. The analyte and internal standard (IS) were isolated from 100ml plasma samples by liquid-liquid extraction (LLE). A Varian 1200l tandem mass spectrometer equipped with an electrospray ionization source was operated in selected reaction monitoring (SRM) mode with the precursor-to-product ion transitions m/z 313.4/138 for granisetron and m/z 270/201 for the IS used for quantitation. The assay exhibited a linear dynamic range of 0.02-20ng/ml for granisetron in human plasma. The lower limit of quantification (LLOQ) was 0.02ng/ml with a relative standard deviati...

Science.gov (United States)

Typing Workshop": [Introductions] [Intro, DNA Basics, and Historical Perspective] [DNA Extraction] [Validation and QA/QC] [DNA Quantitation] [PCR Amplification] [STR Loci and Kits]...

KoreaScience (Korea)

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UK PubMed Central (United Kingdom)

The genes which encode glycosomal glyceraldehyde-phosphate dehydrogenase (gGAPDH) of Trypanosoma cruzi are arranged as a tandemly repeated pair on a single chromosome and are identical at the level...Full Text Available

UK PubMed Central (United Kingdom)

BackgroundYersinia pestis, the agent of plague, is a young and highly monomorphic species. Three biovars, each one thought to be associated with the last three Y....Full Text Available

UK PubMed Central (United Kingdom)

In this study we have profiled the free sterol content of cerebrospinal fluid by a combination of charge tagging and liquid chromatography-tandem mass spectrometry. Surprisingly, the most abundant cholesterol...Full Text Available

UK PubMed Central (United Kingdom)

Purpose: Reverse helical cone-beam computed tomography (CBCT) is a scanning configuration for potential applications in image-guided radiation therapy in which an accurate anatomic image...Full Text Available

Science.gov (United States)

The stability of a symmetric tandem mirror with hot-electron end plugs against low-frequency modes is investigated using a model first proposed by Baldwin. The Baldwin instability, which is due to the coupling of a negative-energy mode in the plug and a shear Alfven mode in the center cell, is confirmed. It is found that this instability is confined to small ranges of value of L/sub c/, the length of the center cell. Under modest restriction of parameters, the unstable regions in L/sub c/ impose little limitation on L/sub c/.

UK PubMed Central (United Kingdom)

A two-step polymerase chain reaction (PCR) procedure with two nested pairs of primers specific for the yadA gene of Yersinia enterocolitica was developed. The PCR assay identified all common pathogenic...Full Text Available

Science.gov (United States)

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...

British Library Electronic Table of Contents (United Kingdom)

The perceptual center (P-center) is fundamental to the timing of heterogeneous event sequences, including music and speech. Unfortunately, there is currently no comprehensive and reliable model of P-centers in acoustic events, so P-centers must instead be measured empirically. This study reviews existing measurement methods and evaluates two methods in detail?the rhythm adjustment method and a new method based on the phase correction response (PCR) in a synchronous tapping task. The two methods yielded consistent P-center estimates and showed no evidence of P-center context dependence. The PCR method appears promising because it is accurate and efficient and does not require explicit perceptual judgments. As a secondary result, the magnitude of the PCR is shown to vary systematically with ...

British Library Electronic Table of Contents (United Kingdom)

A novel PCR based assay was devised to specifically detect contamination of any Salmonella serovar in milk, fruit juice and ice-cream without pre-enrichment. This method utilizes primers against hilA gene which is conserved in all Salmonella serovars and absent from the close relatives of Salmonella. An optimized protocol, in terms time and money, is provided for the reduction of PCR contaminants from milk, ice-cream and juice through the use of routine laboratory chemicals. The simplicity, efficiency (time taken 3-4 h) and sensitivity (to about 5-10 CFU/ml) of this technique confers a unique advantage over other previously used time consuming detection techniques. This technique does not involve pre-enrichment of the samples or extensive sample processing, which was a pre-requisite in mos...

British Library Electronic Table of Contents (United Kingdom)

Evidence suggests that insertion of the IS6110 element is not without consequence to the biology of Mycobacterium tuberculosis complex strains. Thus, mapping of multiple IS6110 insertion sites in the genome of biomedically relevant clinical isolates would result in a better understanding of the role of this mobile element, particularly with regard to transmission, adaptability and virulence. In the present paper, we describe a versatile strategy, referred to as GL-PCR, that amplifies IS6110-flanking sequences based on the construction of a genomic library. M. tuberculosis chromosomal DNA is fully digested with HincII and then ligated into a plasmid vector between T7 and T3 promoter sequences. The ligation reaction product is transformed into Escherichia coli and selective PCR amplification...

UK PubMed Central (United Kingdom)

Borrelial infection may manifest with a wide range of clinical signs, and in many cases, microbiological findings are essential for a proper diagnosis. This study included 48 patients with a working...Full Text Available

UK PubMed Central (United Kingdom)

Clostridium difficile (C. difficile) is a common causative agent of pseudomembranous colitis (PMC). C. difficile-associated diarrhea (CDAD) ranges...Full Text Available

UK PubMed Central (United Kingdom)

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

UK PubMed Central (United Kingdom)

Epidermal growth factor receptor (EGFR) kinase domain mutations hyperactivate the kinase and confer kinase addiction of the non-small-cell lung cancer (NSCLC) tumor...Full Text Available

UK PubMed Central (United Kingdom)

Striped bass (Morone saxatilis) in the Chesapeake Bay are currently experiencing a very high prevalence of mycobacteriosis associated with newly described Mycobacterium...Full Text Available

UK PubMed Central (United Kingdom)

BackgroundThe genus Vibrio is a diverse group of Gram-negative bacteria comprised of 74 species. Furthermore, the genus has and is expected to continue expanding...Full Text Available

Energy Technology Data Exchange (ETDEWEB)

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 PCR products. This sequence represents a ...

UK PubMed Central (United Kingdom)

The methanogenic community in hydrothermally active sediments of Guaymas Basin (Gulf of California, Mexico) was analyzed by PCR amplification, cloning, and sequencing of methyl coenzyme M reductase...Full Text Available

UK PubMed Central (United Kingdom)

The vaginal bacterial microbiota of 19 premenopausal women was examined by PCR-denaturing gradient gel electrophoresis (DGGE) and sequencing of the V2-V3 region of the 16S rRNA gene. Ten of the women...Full Text Available

UK PubMed Central (United Kingdom)

CTX-M extended-spectrum β-lactamases (ESBLs) are increasingly prevalent worldwide among Escherichia coli bacteria, mostly in community-acquired urinary tract infections. Finding...Full Text Available

British Library Electronic Table of Contents (United Kingdom)

Percutaneous coronary revascularisation [PCR] improves angina and health related quality of life [HRQOL] compared to standard medical therapy. It is unknown whether PCR has the same benefits for patients with a history of CABG. Over a period of 5?years, we assessed HRQOL of patients undergoing PCR using Part 1 of the Nottingham Health Profile [NHP] at baseline 3, 12 and 24?months. We compared HRQOL after PCR in 255 patients with CABG to 2680 without. There were more males [81.1% v 69.6% p?=?0.002] and older patients [mean age 60.1?years v. 58.0 p?=?0.03] in CABG group. Perceived HRQOL improved at 24?months for pain, energy and emotional reaction but the improvement was less in the CABG group. However, mean NHP scores at 24?months for those with CABG had returned to baseline levels for slee...

UK PubMed Central (United Kingdom)

The primary mode of transmission of Helicobacter pylori, a human pathogen carried by more than half the population worldwide, is still unresolved. Some epidemiological data suggest...Full Text Available

UK PubMed Central (United Kingdom)

AIMS: To evaluate the sensitivity of the Roche Cobas, Roche Amplicor plate kit, ligase chain reaction (LCR), and an in house polymerase chain reaction (PCR) by titration of purified elementary bodies...Full Text Available

UK PubMed Central (United Kingdom)

Objective: Creatine and phosphocreatine (PCr) are important compounds in the normal energy metabolism of muscle. Recently, it has been shown that dietary creatine (5 to 20 g/day)...Full Text Available

UK PubMed Central (United Kingdom)

SummaryDNA-based tests are increasingly being used to predict a blood group phenotype. This is possible because genes encoding 29 of the 30 blood group systems have been cloned and sequenced,...Full Text Available

UK PubMed Central (United Kingdom)

Arbuscular mycorrhizal fungi (AMF) are obligate biotrophs, known to play an important role in ecological processes. Conventional light microscopy is the most common method used to detect their presence...Full Text Available

British Library Electronic Table of Contents (United Kingdom)

Problems experienced during Floating Production, Storage and Offloading (FPSO) tandem offloading operations were investigated. The aim of this research was to improve the reliability of such systems, and it needed a means to assess them. Time-domain simulation and analysis of offloading systems was performed using the multi-body mooring software ARIANE 7.0. Hydrodynamic interaction between the vessels was considered. The responses of the offloading system in different loading cases, different parameters of offloading hawsers and the effects of challenging environmental conditions were calculated. There was a focus on the problems of relative motion between the two bodies and its effects on the intensity of hawser forces. Minimum relative distance, maximum relative headings and maximum tens...

Energy Technology Data Exchange (ETDEWEB)

In this paper, research on pass schedule of tandem cold mill (TCM) is carried out. According to load (reduction, rolling force, motor power) balance, non-linear equations set with variables of inter-stand thickness is constructed. The pass schedule optimization is carried out by solving the non-linear equations set. As the traditional method, the Newton-Raphson method is used for solving the non-linear equations set. In this paper a new simple method is brought up. On basis of the monotone relations between thickness and load, the inter-stands thickness is adjusted dynamically. The solution of non-linear equations set can be converged by iterative calculation. This method can avoid the derivative calculation used by traditional method. So, this method is simple and calculation speed is high. It is suitable for on-line control. (author)

British Library Electronic Table of Contents (United Kingdom)

Phenytoin (PHT), phenobarbital (PHB), lamotrigine (LTG), and topiramate (TPM) are some of the most widely used antiepileptic drugs (AEDs). Monitoring of their concentrations in serum is important for the treatment of epilepsy. A reference measurement procedure (RMP) for certification of PHT, PHB, LTG, and TPM in serum has been developed and critically evaluated. Isotopically labeled compounds of PHT, PHB, LTG, and TPM are used as internal standards for the four AEDs. The four drugs and their respective labeled internal standards are simultaneously extracted from serum using solid-phase extraction prior to reversed-phase liquid chromatography?tandem mass spectrometry (LC-MS/MS). Chromatographic separation was performed using a C18 column. Electrospray ionization (ESI) in the positive ion mo...

Energy Technology Data Exchange (ETDEWEB)

Changes in the phosphate metabolism of the transplanted SCCVII (squamous cell carcinoma VII) tumor were studied before and after irradiation using in vivo [sup 31]P-magnetic resonance spectroscopy (MRS). Phosphocreatine (PCr), [gamma]-, [alpha]-, [beta]-ATP, inorganic phosphate (Pi), phosphomonoester (PME), and phosphodiester (PDE) were detected in [sup 31]P-MR spectra of tumor tissues. The curve fitting method was employed for calculating each peak of [sup 31]P-MR spectra (PCr, [beta]-ATP, Pi, PME, PDE). The values of PCr, PCr/Pi, [beta]-ATP/Pi and pH decreased in proportion to the growth of tumor, whereas that of Pi, PME and PDE increased. But the changes of [beta]-ATP were minimal in degree. Based on the fact that the ratio of PCr/Pi highly correlated with tumor volume and showed the most remarkable change among various parameters, it might be the useful index for the tumor ...

Energy Technology Data Exchange (ETDEWEB)

Lawrence Livermore National Laboratory (LLNL) has implemented a computer control system for operation of an FN tandem accelerator. The control software utilized is the Thaumaturgic Automated Control Logic (TACL) written by the Continuous Electron Beam Accelerator Facility and co-developed with LLNL. Details of the design philosophy, hardware configuration, control software, and special control algorithms will be presented. 2 refs., 4 figs.

CERN Document Server

By using a laser and maser in tandem, it is possible to obtain laser action in the hot exhaust gases involved in heat engine operation. Such a "quantum afterburner" involves the internal quantum states of working gas atoms or molecules as well as the techniques of cavity quantum electrodynamics and is therefore in the domain of quantum thermodynamics. As an example, it is shown that Otto cycle engine performance can be improved beyond that of the "ideal" Otto heat engine.

International Nuclear Information System (INIS)

The high-frequency precessional mode of a hot-electron-stabilized magnetic configuration has previously been shown to be stable in a window of core-plasma mass. Under conditions of frequency matching, the resulting stable negative-energy precessional wave can be destabilized by coupling to positive-energy shear-Alfven waves. Coupling is avoided when the hot-electron precession frequency exceeds the core-plasma ion gyrofrequency.

UK PubMed Central (United Kingdom)

4.5S RNA is a group of RNAs 90 to 94 nucleotides long (length polymorphism due to a varying number of UMP residues at the 3' end) that form hydrogen bonds with poly(A)-terminated RNAs isolated from...Full Text Available

Science.gov (United States)

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

Science.gov (United States)

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 ...

Energy Technology Data Exchange (ETDEWEB)

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

Science.gov (United States)

Xenotropic murine leukemia virus-related virus (XMRV) was discovered in human prostate tumors and later in some chronic fatigue syndrome (CFS) patients. However, subsequent studies have identified various sources of potential contamination with XMRV and other murine leukemia virus (MLV)-related sequences in test samples. Biological and nucleotide sequence analysis indicates that XMRV is distinct from known xenotropic MLVs and has a broad host range and cell tropism including human cells. Therefore, it is prudent to minimize the risk of human exposure to infection by evaluating XMRV contamination in cell lines handled in laboratory research and particularly those used in the manufacture of biological products. Nested DNA PCR assays were optimized for investigating XMRV gag and env sequences in various cell lines, which included MRC-5, Vero, HEK-293, MDCK, HeLa, and A549, that may be used in the development of some vaccines and other cell lines broadly used in ...

British Library Electronic Table of Contents (United Kingdom)

The Pacific white shrimp, Litopenaeus vannamei, is the most important shrimp species in volume in world aquaculture. However, in recent decades, outbreaks of diseases, especially viral diseases, have led to significant economic losses, threatening the sustainability of shrimp farming worldwide. In 2004, Brazilian shrimp farming was seriously affected by a new disease caused by the Infectious myonecrosis virus (IMNV). Thus, disease control based on rapid and sensitive pathogen detection methods has become a priority. In this study, a specific quantitation method for IMNV was developed using real-time PCR with SYBR Green chemistry and viral load of the principal target tissues of chronically infected animals was quantified. The quantitative analysis revealed that mean viral load ranged from ...

British Library Electronic Table of Contents (United Kingdom)

We investigated the presence of XMRV in a cohort of Quebec patients with chronic fatigue syndrome (CFS). DNA was purified from activated peripheral blood mononuclear cells (PBMCs) and PCR was used to detect XMRV gag and env in 72 patients. Anti-XMRV antibodies were searched in sera of 62 patients by Western blot analysis. Attempts to detect XMRV antigens was made, using immunofluorescence with Gag anti-p30 antiserum on activated PBMC from 50 patients. Plasma viremia was measured by RT-PCR on 9 subjects. Finally, detection of infectious virus in 113 CFS subjects was made by co-culture of PHA+IL-2 activated PBMC with human LNCaP carcinoma cells, and by infecting the same susceptible cells with plasma, using a reverse transcriptase (RT) assay as a readout in both experiments. No detection of ...

British Library Electronic Table of Contents (United Kingdom)

SummaryBackgroundCandida albicans is one of the most important etiologic agents causing superficial and deep fungal infections. For prevention of candidiasis, it is important to develop a rapid system that discriminates C. albicans at the strain level.ObjectiveTo develop a system that can identify C. albicans at the strain level.MethodsGenomic DNAs were purified from 179 clinical isolates of C. albicans, and were used as templates for PCR amplification of 25S rDNA and ALT repeats in repetitive sequences (RPSs). PCR products generated from ALT repeats were digested with EcoRI and/or ClaI in order to study the relationships between restriction profiles and amplification profiles.ResultsOne hundred and seventy nine clinical isolates were grouped into genotypes A (92 isolates), B (38 isolates)...

Energy Technology Data Exchange (ETDEWEB)

This paper describes a technique for implementing the ultrasonic inside diameter (ID) creeping wave technique for detection and sizing ID connected defects using a phased array ultrasonic system. The technique uses multiple focal laws to produce the examination modes. The first focal law is designed to create a shear wave nominally at the critical angle for mode conversion to a longitudinal wave at the ID of a part, thus creating a creeping wave. This focal law is focused at the ID to improve sensitivity. The rest of the laws are designed to create tandem sound paths that progress up a vertical surface directly above the focal point of the creeping wave generation point. When a defect on the inner surface is detected with the creeping wave, the height of the defect can be measured from the response of a set of tandem laws without readjusting the position of the probe. Results from standard one-inch long notches of varying depths are presented.

Energy Technology Data Exchange (ETDEWEB)

The Holifield Radioactive Ion Beam Facility (HRIBF) now under construction at the Oak Ridge National Laboratory will use the 25-MV tandem accelerator for the acceleration of radioactive ion beams to energies appropriate for research in nuclear physics; negative ion beams are, therefore, required for injection into the tandem accelerator. Because charge exchange is an efficient means for converting initially positive ion beams to negative ion beams, both positive and negative ion sources are viable options for use at the facility; the choice of the type of ion source will depend on the overall efficiency for generating the radioactive species of interest. A high-temperature version of the CERN-ISOLDE positive ion source has been selected and a modified version of the source designed and fabricated for initial use at the HRIBF because of its low emittance, relatively high ionization efficiencies and species versatility, and because it has been ...

Energy Technology Data Exchange (ETDEWEB)

A brief description of a method for producing relatively intense molecular negative ion beams for the difficult Group IIA elements is given which offers considerable improvement in terms of source operation and beam intensity stability over other methods conventionally utilized. It is particularly suited for use in cesium plasma sources such as the Aarhus geometry and axial geometry versions of the source. The method utilizes H/sub 2/ source feed gas for the production of a hydrogen-rich plasma discharge which sputters a negatively biased probe made of elemental or copper alloy material. Negative ion beams of MgH/sub 3//sup -/>=12 ..mu.. A have been realized during routine operation of the 25 MV tandem accelerator. Negative ion beam intensity data, typical source operational parameters, and examples of mass spectra associated with their production are given. Interesting intermetallic molecular negative ion beams consisting of the particular Group IIA element and ...

Energy Technology Data Exchange (ETDEWEB)

The general usefulness of neutron activation analysis (NAA) for samples containing the platinum group elements (PGE) and Au, either as major or trace constituents, is discussed. Charged particle activation is shown to be a viable or complementary alternative. Proton (6-10 MeV) and alpha particle (9-15 MeV) beams, produced in a tandem Van de Graaff accelerator, have been established to be the most effective choice. By taking advantage of the Coulomb barrier effect and the fact that many charged particle induced reactions have relatively large threshold values, the same instrumental technique can be applied, with minor modifications, to a wide range of materials.

Energy Technology Data Exchange (ETDEWEB)

Two sources of negative ions are described. An rf source produces up to 14 ..mu..A beams of He/sup -/ by charge exchange in Rb vapor. The other Source of Negative Ions by Cesium Sputtering (SNICS) produces a wide variety of negative ion beams in the ..mu..A range. Two important features of SNICS are its simple, compact construction and its very good beam emittance (2 to 4..pi..mm mrad MeV/sup 1/2/). Both sources have lifetimes >200 hours and they are used extensively on the Wisconsin EN tandem.

British Library Electronic Table of Contents (United Kingdom)

We operate a new NEC 250kV single-stage accelerator mass spectrometer (SSAMS) next to our established 5MV tandem. This permits good comparison of 14C-AMS and challenges SSAMS performance. Initial SSAMS ion-optical deficiencies have been addressed by shimming the injection magnet and 3permil 14C/13C measurement with background limited by sample chemistry is routine. Higher-precision analysis is also achievable. A problematic measurement interference remains, however. The small spectrometer was procured to add measurement capacity, and for experiments with positive ions, but is now the preferred instrument for both convenience and measurement quality.

Energy Technology Data Exchange (ETDEWEB)

We report on multijunction GaInP/GaAs photovoltaic cells with total-area efficiencies of 29.5% at one-sun concentration and air mass (AM) 1.5 global and 25.7% one-sun, AM0. These values represent the highest efficiencies achieved by any solar cell under these illumination conditions. Three key areas in this technology are identified and discussed: the grid design, front surface passivation of the top cell, and bottom surface passivation of both cells. Aspects of cell design related to its operation under different solar spectra and under concentration are also discussed.

British Library Electronic Table of Contents (United Kingdom)

Purpose Study the radiolysis of solid-state metoclopramide hydrochloride at various absorbed doses. Elucidate the structure of the degradation products to gain information on the radiolysis mechanisms. Methods Solid-state metoclopramide samples were irradiated at several doses with gamma rays and high-energy electrons to evaluate the influence of the dose rate. High-performance liquid chromatography with a diode array detector was used to measure the chemical potency as a function of the absorbed dose and to quantify the degradation products. The characterization of degradation products was performed by liquid chromatography/atmospheric pressure chemical ionization/tandem mass spectrometry. Results The degradation of solid-state metoclopramide after irradiation was negligible. No qualitati...

Energy Technology Data Exchange (ETDEWEB)

Eukaryotic genomes encode a zinc finger protein (ZPR1) with tandem ZPR1 domains. In response to growth stimuli, ZPR1 assembles into complexes with eukaryotic translation elongation factor 1A (eEF1A) and the survival motor neurons protein. To gain insight into the structural mechanisms underlying the essential function of ZPR1 in diverse organisms, we determined the crystal structure of a ZPR1 domain tandem and characterized the interaction with eEF1A. The ZPR1 domain consists of an elongation initiation factor 2-like zinc finger and a double-stranded {beta} helix with a helical hairpin insertion. ZPR1 binds preferentially to GDP-bound eEF1A but does not directly influence the kinetics of nucleotide exchange or GTP hydrolysis. However, ZPR1 efficiently displaces the exchange factor eEF1B from preformed nucleotide-free complexes, suggesting that it may function as a negative regulator of eEF1A activation. Structure-based mutational and ...

Energy Technology Data Exchange (ETDEWEB)

Chapter 5, describes some of the most important molecular methods used in the study of chromosome structure and function. The methods discussed include fragmentation of DNA, cloning, flow cytometry and chromosome sorting, is situ hybridization, polymerase chain reaction (PCR), and yeast artificial chromosomes (YACs). 18 refs., 3 figs., 1 tab.

International Nuclear Information System (INIS)

The development of leukemia and thyroid cancer is characterized by activation of the abl oncogene and ret oncogene, respectively. In order to clarify the relationship between these gene aberrations and radiation, the pro-myelogenous leukemia-derived cell line HL60 and the thyroid cancer-derived cell line 8505C, were irradiated in vitro with 100Gy of X-rays. RNA was then extracted from 10"8 cells of the respective cell lines and examined by the reverse transcription PCR method for rearrangements of abl and ret genes. Five kinds of positive bands were observed in the HL-60 cells irradiated with 100Gy of X-ray. Similarly, six positive bands were also observed in the 8505C cells irradiated with 100Gy. In vitro X-irradiation activation of oncogenes found in radiation induced cancers imply that gene rearrangement by X-rays is involved in the development of malignant tumors. Furthermore, in an experiment to detect radiation effects in A-bomb survivors, RNA was extracted ...

UK PubMed Central (United Kingdom)

To determine if white-tailed deer may serve as a reservoir host for Borrelia lonestari, we used a nested PCR for the Borrelia flagellin gene to evaluate blood samples...Full Text Available

British Library Electronic Table of Contents (United Kingdom)

SummaryA laboratory-based surveillance study was conducted from January 2007 to May 2010 in San Martino Tertiary Referral Hospital in Genoa, Italy in which the molecular epidemiology of multidrug-resistant Acinetobacter baumannii was investigated in the five intensive care units (ICUs). A total of 53 A. baumannii strains were isolated from patients admitted to ICUs (69.8%) and to other epidemiologically linked hospital wards (30.2%) and were genotyped by repetitive extragenic palindromic polymerase chain reaction (REP-PCR), multilocus sequence typing (MLST) and adeB sequence typing. REP-PCR fingerprinting analysis, MLST and adeB typing results were well correlated and allowed us to classify strains causing epidemic events into three major epidemic clones: A (REP-I/ST4, adeB-STII genotype) ...

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Aims: The present study explored the integrative effect of genes encoding methadone pharmacokinetic and pharmacodynamic pathways on methadone maintenance doses in Han Chinese Patients. Materials & methods: Genomic DNA was extracted from 321 opioid-dependent patients and 202 healthy controls, and realtime-PCR and PCR-RFLP were conducted to determine the genotypes. Results: Pair-wise comparisons revealed that carriers of the variants ABCB1 3435C>T or CYP2B6 516G>T alleles were more likely to require a higher methadone dose than noncarriers (both p G or 939C>T allele had a two-fold chance of requiring a lower methadone dose than noncarriers (p = 0.001). Proportional odds regression with adjustment of cofactors demonstrated that ...

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MicroRNAs (miRNAs) are small non-coding RNAs that play vital roles in plant abiotic stress responses via cleavage or translational inhibition of their target mRNAs. Populus euphratica is a typical stress-resistant sessile organism that grows in desert areas. Here, we identified sequences of 12 miRNA precursors from 11 families and 13 mature miRNAs from 12 families by PCR amplification in P. euphratica. To detect expression differences in mature miRNAs and their precursors under dehydration and high salinity shock in P. euphratica, we examined 14 miRNA precursors from 13 miRNA families and 17 mature miRNAs from 17 miRNA families using the SYBR Green RT?PCR assay. This is the first report of expression profiles for both precursor and mature miRNAs in P. euphratica. By profiling both the matu...

International Nuclear Information System (INIS)

Several different aspects of the influence of the target backing on in-beam electron spectra following compound nuclear reactions induced by accelerated ions at tandem energies irradiating backed targets are discussed in detail. This discussion is illustrated by a few typical examples, such as "1"2C"5"+ and "3"1P"1"0"+ beams at 4 MeV/u bombarding Sn(+Be), Sn(+Au), Pb(+C) backed targets. Moreover, the relative influence of electron backscattering, electron Doppler shift and Doppler broadening as well as #delta#-electron emission on the low energy electron spectra (E_e#<=#100 keV) obtained under such conditions are investigated in the frame of the available experimental data. (orig.).

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As a consequence of the close integration of cattle into the food chain of humans, forensically relevant cases involving cattle (Bos taurus) DNA analysis are common. However, scientific publications reporting the information content of the commonly used bovine short tandem repeat (STR) loci remains scarce. Population studies were performed for 16 polymorphic STR loci (BM1818, BM1824, BM2113, CSRM60, CSSM66, ETH3, ETH10, ETH225, HAUT27, ILSTS006, INRA023, SPS115, TGLA53, TGLA122, TGLA126, and TGLA227) including 4,162 randomly selected cattle representing 20 distinct breeds. The power of parental exclusion, expected and observed heterozygosity, probability of identity, and non-amplifying (?null?) allele frequencies were calculated. Major differences existed in the information content between...

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BACKGROUND AND PURPOSE It is well established that cytochrome P450 2J (CYP2J) enzymes are expressed preferentially in the heart, and that ebastine is a substrate for CYP2J, but it is not known whether ebastine is metabolized in myocardium. Therefore, we investigated its pharmacokinetics in the rat isolated perfused heart. EXPERIMENTAL APPROACH Rat isolated hearts were perfused in the recirculating mode with ebastine for 130-min. The concentrations of ebastine and its metabolites, hydroxyebastine and carebastine, were measured using liquid chromatography with a tandem mass spectrometry. The data were analysed by a compartmental model. The time course of negative inotropic response was linked to ebastine concentration to determine the concentration-effect relationship. KEY RESULTS Ebastine w...

Energy Technology Data Exchange (ETDEWEB)

An energy saving dc-dc converter circuit is disclosed having two energy efficient means which operate in tandem, an energy conserving means and a voltage doubling means. These energy efficient means are applied in combination with elements commonly found in dc-dc converter circuits, namely an ac voltage generator, a transformer for stepping up the generated ac voltage, and means for storing the converted dc voltage. The energy conserving means is connected to the dc voltage storage means. It comprises a resettable inhibit circuit which cuts off the provision of dc voltage for conversion for a predetermined interval when the output of the converter exceeds a predetermined level. The voltage doubling means is reponsive to outputs of the inhibit circuit of the energy conserving means and the ac voltage generator. It provides a phase inverted waveform of the generated ac voltage on one of two leads to the ac voltage step-up transformer.

Science.gov (United States)

Synthetic efforts toward the homoerythrina alkaloids 1-3 are described. Two separate model systems guided the pivotal [3 + 2] azomethine ylide cycloaddition cascade to form the A-C rings of these alkaloids. The cycloaddition precursors 63 and 68, prepared in nine and ten steps, respectively, from alkyne 47, each contain an enolizable ketone, a tethered electrophile, and an electron-poor dipolarophile. Heating 63 and 68 with the stannyl amine 17 generated demethoxyschelhammeridine 65 and demethoxyschelhammericine 70, the products of intramolecular azomethine ylide cycloadditions. Subsequent attempts to install the C-3 methoxy group of 1-3 are also described. PMID:17465572

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The real-time polymerase chain reaction (PCR) quantification of several vaginal bacterial groups in healthy women and patients developing asymptomatic bacterial vaginosis (BV) and candidiasis (CA) was performed. Statistical analysis revealed that the BV condition is characterised by a great variability among subjects and that it is associated with a significant increase of Prevotella, Atopobium, Veillonella and Gardnerella vaginalis, and a drop in Lactobacillus. On the contrary, the vaginal microflora of healthy women and patients developing CA was found to be homogeneous and stable over time.

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IntroductionBreast cancer is the most diagnosed and second leading cause of cancer deaths in the U.S. female population. An estimated 5 to 10 percent of all breast cancers are inherited, caused by mutations in the breast cancer susceptibility genes (BRCA1/2). As many as 90% of all mutations are nonsense mutations, causing a truncated polypeptide product. A popular and low cost method of mutation detection has been the protein truncation test (PTT), where target regions of BRCA1/2 are PCR amplified, transcribed/translated in a cell-free protein synthesis system and analyzed for truncated polypeptides by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography. We previously reported a novel High Throughput Solid-Phase PTT (HTS-PTT) based on an enzyme-linked immunosorbent assay (ELISA) format that eliminates the need for radioactivity, SDS-PAGE and subjective interpretation of the results. Here, we report the next generation HTS-PTT ...

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By polymerase chain reaction with arbitrary primer (AP-PCR), the possibility of transmission of genome instability to somatic cells of the offspring (F_1 generation) from male parents of mice exposed to chronic low-dose #gamma#-radiation was studied. Male mice 15 days after exposure to 10-50 cGy were mated with unirradiated females. Biopsies were taken from tale tips of two month-old mice progeny for DNA separation. Primer in the AP-PCR was 20-mer oligonucleotide flanking the micro-satellite locus Atplb2 on chromosome 11 of the mouse. Comparative analysis of individual fingerprints of AP-PCR products on DNA-templates from the offspring of irradiated and unirradiated male mice revealed an increased variability of micro-satellite-associated sequences in the genome of the offspring of males exposed to 25 and 50 cGy. DNA-fingerprints of the offspring of male mice exposed to chronic irradiation doses 10 and 25 cGy. 15 days ...

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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 continued multiplication of the affected cell. Ionizing radiation-induced DNA rearrangements were detected in ...

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The Holifield Radioactive Ion Beam Facility now under construction at the Oak Ridge National Laboratory will use the 25 MV tandem accelerator for the acceleration of radioactive ion beams to energies appropriate for research in nuclear physics; negative ion beams are, therefore, required for injection into the tandem accelerator. Because charge exchange is an efficient means for converting initially positive ion beams to negative ion beams, both positive and negative ion sources are viable options for use at the facility. The choice of the type of ion source will depend on the overall efficiency for generating the radioactive species of interest. Although direct-extraction negative ion sources are clearly desirable, the ion formation efficiencies are often too low for practical consideration; for this situation, positive ion sources, in combination with charge exchange, are the logical choice. The high-temperature version of the CERN-ISOLDE ...

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Radioactive ion beams (RIBs) of short-lived isotopes of fluorine are in demand for investigating astrophysical phenomena related to the hot CNO cycle and rp processes responsible for stellar nucleosynthesis. Since negative ion beams are required for injection into tandem electrostatic accelerators, such as the 25 MV tandem accelerator used for post acceleration of RIBs for the Holifield Radioactive Ion Beam Facility (HRIBF) research program at the Oak Ridge National Laboratory (ORNL), efficient, direct-formation F{sup -} ion sources are highly desirable for RIB applications involving this type of post-accelerator. We have conceived and evaluated a direct extraction F{sup -} source for potential RIB applications which is predicated on the reverse polarity operation of a positive electron-beam-plasma target/ion source (EBPTIS) while simultaneously feeding fluorine rich compounds and Cs vapor into the source. The source is found to operate in two ...

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Purpose To study the activation and expression of vascular (aorta and small mesenteric arteries) potassium channels during septic shock with or without modulation of the NO pathway. Methods Septic shock was induced in rats by peritonitis. Selective inhibitors of vascular KATP (PNU-37883A) or BKCa [iberiotoxin (IbTX)] channels were used to demonstrate their involvement in vascular hyporeactivity. Vascular response to phenylephrine was measured on aorta and small mesenteric arteries mounted on a wire myograph. Vascular expression of potassium channels was studied by PCR and Western blot, in the presence or absence of 1400W, an inducible NO synthase (iNOS) inhibitor. Aortic activation of the transcriptional factor nuclear factor-kappaB (NF-?B) was assessed by electrophoretic mobility shift as...

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

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The antimicrobial peptide scygonadin (Scy) was first isolated from the gonad of Scylla serrata and its gene is predominantly expressed in the ejaculatory duct of adult males. Thus, its function was predicted to be associated with reproductive immunity, but this is still unclear and needs further investigation. In our study, the expression pattern of Scy at different developmental stages of both male and female S. paramamosain was investigated, so that the potential function of this peptide could be examined. Using real-time quantitative PCR, Scy mRNA transcripts were demonstrated obviously in the vulnerable embryos and larvae-zoea I but very weakly detected in the larvae-zoea III, megalops and juveniles. The gene expression pattern showed a decreasing trend during the early developmental s...

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AbstractBackground: A defective innate immune response may contribute to the pathogenesis of Crohn's disease (CD) and ulcerative colitis (UC). Employing a global gene expression analysis, this study was aimed at identifying specifically regulated genes within the epithelial compartment in inflammatory bowel disease (IBD). Methods: The epithelial fraction of human ileal mucosa samples from surgical specimens was obtained by laser microdissection. Gene expression was examined by global expression profiling (n = 18, Affymetrix), quantitative reverse-transcription polymerase chain reaction (RT-PCR) (n = 35), immunoblot analysis (n = 9), and immunohistochemistry (n = 25). Results: Global expression profiling revealed a pronounced downregulation of the retinoic acid-inducible gene I (RIG-I) with...

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Abstract Strains of Pseudomonas syringae are effective in controlling postharvest diseases of citrus fruits, and antagonistic activity has been correlated with in vitro production of lipodepsipeptides. Additionally, biocontrol agents can induce a range of defence mechanisms of resistance in citrus tissue that result in a broad spectrum of metabolic modifications, such as systemic acquired resistance, induced systemic resistance and production of reactive oxygen species. The aim of this study was to evaluate the expression of syringomycin (syrB1) and syringopeptin (sypA) synthetase genes from P.syringae pv. syringae biocontrol strains in vitro on different culture media and in vivo on citrus fruits (Citrus sinensis cv. Tarocco) during the interaction with Penicillium digitatum by quantitati...

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High-level expression of mitotic arrest defective protein 2 (MAD2), a central component of the spindle assembly checkpoint, has been observed in a variety of human malignancies. Aim of the present study was to observe the expression of MAD2 in human non-small-cell lung cancer (NSCLC) and explore its clinicopathologic significance and evaluate MAD2 expression as a prognostic marker. MAD2 transcript was found to be overexpressed in the great majority of lung cancers by semi-quantitative RT-PCR. A total of 358 NSCLCs were analyzed immunohistochemically on tissue microarrays. High-level MAD2 expression was observed in 26.3% (94 of 358 cases), and correlated with male sex (P=0.0002), tumor progression (pT status) (P=0.0009), visceral or parietal pleural invasion (P=0.0151), non-adenocarcinoma, ...

International Nuclear Information System (INIS)

The buoyancy driven convective flow fields are steady circulatory flows which were made between surfaces maintained at two fixed temperatures. They are ubiquitous in nature and play an important role in many engineering applications. Especially, in last decades, natural convection in a close loop or cavity becomes the main issue in the molecular biology for the polymerase chain reaction (PCR). Application of a natural convection can reduce the costs and efforts remarkably. This paper focuses on the sensitivity study of turbulence analysis using CFD for a natural convection in a closed rectangular cavity. Using commercial CFD code, FLUENT, various turbulent models were applied to the turbulent flow. Results from each CFD model will be compared each other in the viewpoints of flow characteristics. This work will suggest the best turbulent model of CFD for analyzing turbulent flows of the natural convection in an enclosure system.

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Abstract Background: G proteins are ubiquitously expressed signal transduction proteins playing a key role in multiple signal transduction pathways. The Gas subunit has been considered as an apoptosis factor. In this study the role of GNAS T393C genotypes of the GNAS gene encoding Gas was analyzed for its influence on the development and progression of prostate cancer. Methods: Genotyping of the GNAS T393C polymorphism in 196 prostate cancer patients and 200 healthy controls was performed by DNA extraction followed by PCR and restriction analysis. Results: We observed no evidence of effects related to GNAS T393C genotype as demonstrated by a comparison of the genotype distribution in prostate cancer patients and healthy controls, the genotype distribution dependent on grade of the primary ...

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BACKGROUND AND PURPOSE Controlling vascular tone involves K+ efflux through endothelial cell small- and intermediate-conductance calcium-activated potassium channels (KCa2.3 and KCa3.1, respectively). We investigated the expression of these channels in astrocytes and the possibility that, by a similar mechanism, they might contribute to neurovascular coupling. EXPERIMENTAL APPROACH Transgenic mice expressing enhanced green fluorescent protein (eGFP) in astrocytes were used to assess KCa2.3 and KCa3.1 expression by immunohistochemistry and RT-PCR. KCa currents in eGFP-positive astrocytes were determined in situ using whole-cell patch clamp electrophysiology. The contribution of KCa3.1 to neurovascular coupling was investigated in pharmacological experiments using electrical field stimulatio...

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DNA methylation is considered as a potential cause of aberrations in regulation of gene expression during carcinogenesis. Therefore, changes in DNA methylation patterns may be targets for chemoprevention. In the present study, we investigated effects of all-trans retinoic acid (ATRA), vitamin D3, and resveratrol alone and in combination with adenosine analogues: 2-chloro-2'-deoxyadenosine (2CdA) and 9-beta-d-arabinosyl-2-fluoroadenine (F-ara-A), on methylation and expression of retinoic acid receptor beta 2 (RARbeta2) in MCF-7 and MDA-MB-231 breast cancer cell lines. Alterations in methylation and expression levels after treatment of cells with the tested compounds were evaluated by methylation-sensitive restriction analysis (MSRA) and real-time PCR, respectively. RARbeta2 promoter in the ...

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Abstract To investigate the link between the functionality and the diversity of microbial communities under strong selective pressure from pollutants, two types of mesocosms that simulate natural attenuation and phytoremediation were generated using soil from a site highly contaminated with jet fuel and under air-sparging treatment. An increase in the petroleum hydrocarbon concentration from 4900 to 18-500-mg-kg-1-dw soil simulated a pollutant rebound (postremediation pollutant reversal due to residual contamination). Analysis of soil bacterial communities by denaturing gradient gel electrophoresis of PCR-amplified 16S rRNA gene fragments showed stronger changes and selection for a phylogenetically diverse microbial population in the mesocosms with pollutant-tolerant willow trees. Enumerat...

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Phytoremediation has been identified as a potentially environmentally friendly and cost effective technique for the treatment of contaminated soil. However, phytoremediation has an unknown mechanism. In this study, we focus on the effects of the cultivation of Italian ryegrass on the soil microbes collected at oil showings, which were expected to have a variety of crude oil degradable microorganisms. We evaluated the number of crude oil degradable microorganism, microbial activity, microflora using the PCR-DGGE method and the change in the concentration of crude oil in the soil. The results indicated that the microflora was affected by the cultivation of Itarian ryegrass, and that the microbial activity and the number of crude oil degradable microorganisms were also improved by the cultivation. Moreover, the concentration of crude oil in the rhizosphere soil decreased significantly when compared to the uncultivated soil. These results suggested that cultivation ...

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

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

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Abstract Purpose: The aim of this in vitro study was to compare the marginal adaptation of a pressed ceramic material, when used with and without a metal substructure, to a traditional feldspathic porcelain-fused-to-metal restoration with a porcelain butt margin. Materials and Methods: A maxillary central incisor typodont tooth was prepared with a 1.5 mm 360degree shoulder with rounded internal line angle, and 30 polyether impressions were made. Dies were poured in type IV dental stone, and 30 restorations were fabricated: 10 metal ceramic restorations (MCR) with porcelain butt joints, 10 pressed to metal restorations (PTM), and 10 all-ceramic restorations (PCR). All restorations were evaluated on their respective dies at 45x magnification using an Olympus SZX-12, measurements of the margi...

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We determined the chromosomal localization and structure of the gene encoding human type II inosine 5{prime}-monophosphate dehydrogenase (IMPDH, EC 1.1.1.205), an enzyme associated with cellular proliferation, malignant transformation, and differentiation. Using polymerase chain reaction (PCR) primers specific for type II IMPDH, we screened a panel of human-Chinese hamster cell somatic hybrids and a separate deletion panel of chromosome 3 hybrids and localized the gene to 3p21.1{yields}p24.2. Two overlapping yeast artificial chromosome clones containing the full gene for type II IMPDH were isolated and a physical map of 117 kb of human genomic DNA in this region of chromosome 3 was constructed. The gene for type II IMPDH was localized and oriented on this map and found to span no more than 12.5 kb.

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A multicusp magnetic field plasma surface ion source, normally used for H/sup -/ ion beam formation, has been utilized for the generation of high intensity, pulsed, heavy negative ion beams suitable for a variety of uses including tandem electrostatic accelerator/synchrotron injection applications. Sputter probe voltage limited total ion currents of 5.5, 8.2, 5.1 and 4.5 mA (peak intensity) have been produced from Au, Cu, Ni and CuO sputter probes, respectively. The mass distributions of these ion beams are found to be dominated by Au/sup -/, Cu/sup -/, Ni/sup -/ and O/sup -/ atomic species, respectively. The source offers the interesting prospect of providing cw negative ion beams at mA intensity levels of the commonly used semiconducting material dopants (e.g. B/sup -/, P/sup -/, As/sup -/ and Sb/sup -/) as well as O/sup -/ for isolation barrier formation. Illustrative examples of intensity versus time and the mass distribution of ion beams extracted from a Ni ...

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In this study, the nearly complete sequence (14,519 bp) of the mitochondrial DNA (mtDNA) of the entomophagous endoparasite Xenos vesparum (Insecta: Strepsiptera) is described. All protein coding genes (PCGs) are in the arrangement known to be ancestral for insects, but three tRNA genes (trnA, trnS(gcu), and trnL(uag)) have transposed to derived positions and there are three tandem copies of trnH, each of which is potentially functional. All of these rearrangements except for that of trnL(uag) is within the short span between nad3 and nad4 and there are numerous blocks of unassignable sequence in this region, perhaps as remnants of larger scale predisposing rearrangements. X. vesparum mtDNA nucleotide composition is strongly biased toward As and Ts, as is typical for insect mtDNAs. There is also significant strand skew in the distribution of these nucleotides, with the J-strand being richer in A than T and in C than G, and the N-strand showing an opposite skew for ...

International Nuclear Information System (INIS)

The purpose of the present study was to examine the in vitro effects of low-dose cadmium (Cd) on developing cortical cells. The cortical cells removed from fetuses (embryonic day 15) were treated with 10 nM of Cd for 24 h. The effects of Cd on dendritic and synaptic development were immunocytochemically observed with anti-microtubule associated protein-2 (MAP2) and anti-synapsin I antibodies, respectively. Administration of Cd suppressed dendritic as well as synaptic development at 10 nM. By two-dimensional gel electrophoresis and liquid chromatography-tandem mass spectrometric (LC/MS/MS) analysis, we identified three proteins with different expression after Cd-treatment; dihydropyrimidinase-related protein 2 (DRP-2/CRMP-2), 14-3-3-epsillon and calmodulin (CaM). Though the number of identified proteins was small, these proteins are known to be involved in neuronal development. The present study demonstrated the morphological effects as well as affected proteins in ...

International Nuclear Information System (INIS)

In experiments designed to detect new mutations affecting hemoglobin, we irradiated the male or female parent in reciprocal crosses of two mouse strains that differ in alleles at the hemoglobin (Hba, Hbb) loci as well as at five other specific loci. Offspring were analyzed for hemoglobin properties (electrophoretic pattern, solubility, crystal pattern), serum albumin differences, erythrocyte lysis, reticulocyte count, and external appearance. Five hemoglobin variants were found among the mutants. In three, the genetic contribution from the irradiated father was not expressed with regard to the #alpha#-chain; one carried a tandem duplication (the first known case in the mouse) involving Hbb; and one probably resulted from double nondisjunction of chromosome 7. The finding that major chromosome aberratios can mimic hemoglobin mutations indicates the need, in similar experiments, to follow F_1 screening with thorough cytogenetic analysis. The variants in which there ...

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The use of excitation system controls, specifically the Power System Stabilizer (PSS), to improve damping of power system oscillations and thus extend steady-state stability limits has been a subject of great interest in recent years. The desire to more fully utilize transmission capacity led to the installation of PSS controls on greater numbers of generating units to maintain stability margins. This paper addresses the tuning and adjustment of PSS controls in a two unit power plant. Both analytical studies and field test confirmations are presented. The practical difficulties of verifying the PSS control loop gain margin by testing are explored. A simplified procedure is outlined. The Lower Colorado River Authority (LCRA), Fayette Power Project (FPP) consists of two 683.7 MVA tandem compound steam turbine driven generators. The excitation systems are of the alternator rectifier type and the PSS control utilizes shaft speed as an input signal. The units must ...

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Described herein are the latest trends in rolling mill facilities toward the 21st. century and new techniques developed by Hitachi Ltd. These cover hot rolling facilities, such as two-stage twin, high crown control-roll cross mill, roll shaping machine and continuous hot rolling systems; cold rolling facilities, such as pickling line-tandem cold mill and other continuous systems; and fine steel rolling facilities, such as roll mill for very small diameter pipes and others to improve productivity and product quality. The other new techniques include systems to pursue reliability and maintainability, in which electric appliances, instruments and computers are integrated, or equipped with built-in large-capacity inverters and drivers. For hot rolling, the above techniques lead to development of a new system named Hi-HOT, which produces high-quality hot coils at a sufficient speed, while keeping rolled steel at a desired temperature. 1 ref., 3 figs.

International Nuclear Information System (INIS)

Electron cyclotron resonance (ECR) ion sources possess several advantages over ion sources conventionally used for injectors of electrostatic accelerators: improved reliability, high efficiency, simplicity, and the capability of generating bright, high-current ion beams. We have adapted a high-current ECR source originally developed as an injector for a CW RFQ proton linac to serve as a source of intense negative-ion beams for the Tandem Accelerator Superconducting Cyclotron (TASCC) facility at Chalk River. The range of ion species of the source has been extended from H1+ alone up to Bi1+, with both gaseous and nonvolatile feeds. Two intense negative-ion beams of He- and O- have been generated so far with the source coupled to a standard charge-exchange canal. We foresee no major problems generating a broad range of negative ions with this system. (author)

International Nuclear Information System (INIS)

Thermoluminescence (TL) dosimetry was used to measure dose effects on the raw stock material of borosilicate container glass from different geographical locations. Effects were studied at times up to 60 days post-irradiation at doses from 0.15 to 20 Gy. The minimum detectable dose using this technique was estimated to be 0.15 Gy which is roughly equivalent to a 24 hr irradiation 1 cm from a 50 ng source of 60Co. Two peaks were identified in the TL glow curve, a relatively unstable peak around 125 C and a more stable peak around 225 C. Differences in TL glow curve shape and intensity were also observed for the glasses from different geographical origins. We investigate radiation induced defects in glass to further develop the technique of intrinsic dosimetry - the measurement of the total absorbed dose received by the walls of a container holding radioactive material. Intrinsic dosimetry is intended to be used as an interrogation tool to provide enhanced pathway information on ...

International Nuclear Information System (INIS)

Chemical analyses were done on the abnormal hemoglobins of the five (101 x SEC)F_1 offspring of X- irradiated adult SEC mice to determine which hemoglobin genes were expressed in each hemoglobin variant. Three offspring of irradiated SEC males did not express either of the two kinds of #alpha#-chains normally found in all SEC mice. The deficient #alpha#-chain synthesis caused these mice to exhibit an #alpha#-thalassemia similar to human #alpha#-thalassemia. Scanning electron microscopy was used to show that many erythrocytes of mice with #alpha#-thalassemia have bizarre shapes; e.g. many erythrocytes appeared flattened or had thorny projections (acanthocytes). One mutant with a tandem duplication of a segment of chromosome 7 (site of locus determining #beta#-chain structure) produced twice as much SEC as 101 #beta#-chain polypeptides. One mutant that probably arose by non-disjunction of chromosome 7's in its unirradiated 101 mother and loss of chromosome 7 from the ...

International Nuclear Information System (INIS)

180 deg magnet chamber is used in Folded Tandem Ion Accelerator for passage and 180 deg bending of ion beam. The chamber is placed between 180 deg terminal magnet (Electro-magnet), which is used for bending, and analysing the beams. Magnet with a particular magnetic field strength bends ions of only specified mass energy product through a precise path. There is also a space limitation in the direction of magnetic field. Both of them require the magnet chamber to be of a close tolerance. Accuracy of center distance between inlet and outlet port of the magnet chamber has to be at par with the concentricity of high energy and low energy beam line. To achieve this we started the fabrication of magnet chamber by following two methods: a) Circular rolling and bending of rectangular tube for 180 deg sector of magnet chamber b) Machining 180 deg sector in two half and welding. By first method we could not achieve the desired result although the method is very promising. ...

Science.gov (United States)

An analytical method was developed for the determination of eleven agrochemicals [abamectin (as B1a), bifenazate, bifenthrin, carfentrazone-ethyl, cymoxanil, hexythiazox, imidacloprid, mefenoxam, pymetrozine, quinoxyfen, and trifloxystrobin] in dried hops. The method utilized polymeric and NH2 solid phase extraction (SPE) column cleanups and liquid chromatography with mass spectrometry (LC-MS/MS). Method validation and concurrent recoveries from untreated dried hops ranged from 71 to 126% for all compounds over three levels of fortification (0.10, 1.0, and 10.0 ppm). Commercially grown hop samples collected from several field sites had detectable residues of bifenazate, bifenthrin, hexythiazox, and quinoxyfen. The control sample used was free of contamination below the 0.050 ppm level for all agrochemicals of interest. The limit of quantitation and limit of detection for all compounds were 0.10 and 0.050 ppm, respectively. PMID:18593182

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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 two sequential affinity chromatography steps to give Sp1 of an estimated 90% homogeneity with 30% yield. In addition, the use of ...

Science.gov (United States)

One of the NASA research activities was to identify, characterize, and simulate a series of technologies that could be used for hydrogen production at NASA Kennedy Space Center (KSC) using locally available sources. This project examined the production of hydrogen from solar energy. To produce hydrogen by water splitting, the operating voltage of conventional photovoltaic (PV) cells cannot supply the overvoltage required. Thus, the objective of this project was to research and develop photoelectrochemical (PEC) cells that can supply the required voltage for water splitting by constructing a multiple bandgap tandem PV cell and a photocatalyst that can be activated by infrared (IR) photons transmitted through the PV cell. The proposed concept is different from conventional PEC water splitting by using multiple band gap combinations. The advantages for this PEC cell concept is that the PV cells are not in contact with the electrolyte solution, thus reducing the ...

Science.gov (United States)

The epidermal compartment is complex and organized into several strata composed of keratinocytes (KCs), including basal, spinous, granular, and cornified layers. The continuous process of self-renewal and barrier formation is dependent on a homeostatic balance achieved amongst KCs involving proliferation, differentiation, and cell death. To determine genes responsible for initiating and maintaining a cornified epidermis, organotypic cultures comprised entirely of stratified KCs creating epidermal equivalents (EE) were raised from a submerged state to an air/liquid (A/L) interface. Compared to the array profile of submerged cultures containing KCs predominantly in a proliferative (relatively undifferentiated) state, EEs raised to an A/L interface displayed a remarkably consistent and distinct profile of mRNAs. Cultures lifted to an A/L interface triggered the induction of gene groups that regulate proliferation, differentiation, and cell death. Next, differentially expressed microRNAs ...

Science.gov (United States)

Twenty autopsy cases with 2009 pandemic influenza A (2009 H1N1) virus infection, performed between August 2009 and February 2010, were histopathologically analyzed. Hematoxylin-eosin staining, immunohistochemistry for type A influenza nucleoprotein antigen, and real-time reverse transcription-PCR assay for viral RNA were performed on formalin-fixed and paraffin-embedded specimens. In addition, the D222G amino acid substitution in influenza virus hemagglutinin, which binds to specific cell receptors, was analyzed in formalin-fixed and paraffin-embedded trachea and lung sections by direct sequencing of PCR-amplified products. There were several histopathological patterns in the lung according to the most remarkable findings in each case: acute diffuse alveolar damage (DAD) with a hyaline membrane (four cases), organized DAD (one case), acute massive intra-alveolar edema with variable degrees of hemorrhage (three cases), neutrophilic ...

DEFF Research Database (Denmark)

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

Science.gov (United States)

A 72-year-old man was exposed to the sarin gas attack in a Tokyo subway on March 20 th, 1995. After exposure, he noticed eye discomfort, chest tightness, headache and weakness of the lower limbs and oropharyngeal muscles. Despite these symptoms, he visited a hot spring on the same day with his family. On March 25 th, his muscle weakness progressed, and a low grade fever appeared. His muscle weakness disappeared 8 days after exposure to sarin, but respiratory failure rapidly developed, necessitating artificial ventilation within four day after hospitalization on March 28th. Chemotherapy with erythromycin, imipenem/cilastatin, and steroid pulse therapy was begu. PCR and culture of sputum collected by bronchofiberscopy were positive for Legionella pneumophila, serogroup I. His respiratory state improved, but subsequent infection with Pseudomonous aeruginosa. Enterobacter cloacae, and Candida tropicalis/glabrata caused his death 71 days after admission. Oropharyngeal ...

Energy Technology Data Exchange (ETDEWEB)

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 16p and mouse 16 is established. 10 refs., ...

British Library Electronic Table of Contents (United Kingdom)

Abstract MicroRNAs (miRNAs) play important roles in global gene regulation. Researchers in recombinant protein production have proposed miRNAs as biomarkers and cell engineering targets. However, miRNA expression remains understudied in Chinese Hamster Ovary cells, one of the most commonly used host cell systems for therapeutic protein production. To profile highly conserved miRNA expression, we used the miRCURY- miRNA array for screening miRNAs in CHO cells. The selection criteria for further miRNA profiling included positive hybridization signals and experimentally validated predicted regulatory targets. On the basis of screening, we selected 16 miRNAs for quantitative RT-PCR profiling. We profiled miR expression in parental CHO DG44 and CHO K1 cell lines as well as four recombinant DG44...

International Nuclear Information System (INIS)

Protocols were standardized for in vitro propagation of several elite and diverse banana accessions using shoot tip explants. Tissue culture raised plants were field planted at multiple locations. Studies were undertaken for the induction of mutations using multiple shoot cultures of six selected cultivars, Shreemanti (AAA), Basrai (AAA), Lal Kela (AAA), Rasthali (AAB), Karibale Monthan (ABB) and a wild diploid (BB). These shoot cultures were irradiated at different doses of gamma rays (0-100 Gy) and subcultured thrice (up to M_1V_3) to separate shimeras, followed by induction of rooting (M_1V_4). In general, the rate of multiplication had a negative association with the dose of gamma rays. Enhanced multiplication of shoots was noticed at lower doses. The proliferation of shoots was arrested beyond 50 Gy and a dose of 70 Gy was completely lethal for all the genotypes studied. The rooted plantlets were hardened in the green house and in the early stages of field growth, a few ...

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This report describes the fiscal 1998 result on development of genome informatics technology. As comparative analysis technique of genes, the combination of electrophoresis and PCR was used. For improvement of the throughput and reproducibility of the technique, module- shuffling primers were used, and the multi(96)-arrayed capillary fragment analyzer was devised. The system detecting SNPs rapidly was also developed successfully. As analysis technology of DNA sequence by use of triple- stranded DNA formation, study was made on construction of long cDNA libraries, selective subtraction of specific sequences from libraries, and the basic technology of homologous cloning. Study was also made on each reaction step of IGCR technique for fast analysis, and specifications of a fluorescence transfer monitor. As modeling technique of genetic sequence information, the simulation model was developed for gene expression regulatory networks during muscle differentiation, and ...

British Library Electronic Table of Contents (United Kingdom)

A new virus species designated as Grapevine leafroll associated virus-Pr (GLRaV-Pr), which is classified in a distinct phylogenetic group of the genus Ampelovirus (Closteroviridae), was recently characterized from Greek grapevine cultivars. Elimination studies of GLRaV-Pr were carried out in two grapevine cultivars, 'Mantilaria' and 'Prevezaniko', co-infected with Grapevine rupestris stem pitting associated virus (GRSPaV, Flexiviridae). Both viruses were detected by nested RT-PCR assays. Virus elimination was achieved by combining in vitro thermotherapy with meristem ( 0.2mm) or shoot tip culture ( 0.5cm). The survival and regeneration rate of meristems was very low. On the other hand, high survival rates were observed in the cultured shoot tips accompanied with high elimination rates for ...

British Library Electronic Table of Contents (United Kingdom)

RNA interference (RNAi) plays an important role in regulating gene expression in eukaryotes. Previously, we generated Arabidopsis and tobacco plants expressing double-stranded RNA (dsRNA) targeting a cotton bollworm (Helicoverpa armigera) P450 gene, CYP6AE14. Bollworms fed on transgenic dsCYP6AE14 plants showed suppressed CYP6AE14 expression and reduced growth on gossypol-containing diet (Mao et?al., in Nat Biotechnol 25: 1307?1313, 2007). Here we report generation and analysis of dsRNA-expressing cotton (Gossypium hirsutum) plants. Bollworm larvae reared on T2 plants of the ds6-3 line exhibited drastically retarded growth, and the transgenic plants were less damaged by bollworms than the control. Quantitative reverse-transcription polymerase chain reaction (RT-PCR) showed that the CYP6AE1...

Science.gov (United States)

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 DNA extraction and bisulfite treatment provides high quality conversion of unmethylated cytosine to uracil ...

British Library Electronic Table of Contents (United Kingdom)

Abstract A bacteriocin-producing Streptococcus bovis strain (HC5) outcompeted a sensitive strain (JB1) before it reached stationary phase (pH 6.4), even though it grew 10% slower and cell-free bovicin HC5 could not yet be detected. The success of bacteriocin-negative S. bovis isolates was enhanced by the presence of another sensitive bacterium (Clostridium sticklandii SR). PCR based on repetitive DNA sequences indicated that S. bovis HC5 was not simply transferring bacteriocin genes to S. bovis JB1. When the two S. bovis strains were coinoculated into minimal medium, bacteriocin-negative isolates predominated, and this effect could be explained by the longer lag time (0.5 vs. 1.5 h) of S. bovis HC5. If the glucose concentration of the minimal medium was increased from 2 to 7 mg mL-1, the e...

British Library Electronic Table of Contents (United Kingdom)

Catalase is an important antioxidant protein which can protect organisms against various oxidative stresses by eliminating hydrogen peroxide. The catalase cDNA of Cristaria plicata@?cpCAT@? was cloned from the haemocytes using degenerate primers by the method of 3' and 5' rapid amplification of cDNA ends PCR. The gene is 4863 bp long and has a total of two introns and three exons. The precise size and location of the introns and exons have been determined. In addition the full-length cDNA of cpCAT contained 2618 bp, The cDNA contained a 5' untranslated region (UTR) of 136 nucleotides, the 3' UTR of 979 bp with a canonical polyadenylation signal sequence AATAAA and a polyA tail, and an open reading frame (ORF) of 1503 bp, encoding 501 amino acid residues with 56.86 kDa predicted molecular w...

Energy Technology Data Exchange (ETDEWEB)

The response of Desulfovibrio vulgaris Hildenborough (DvH),a sulphate-reducing bacterium, to nitrate stress was examined usingquantitative proteomic analysis. DvH was stressed with 105 m M sodiumnitrate(NaNO3), a level that caused a 50 percent inhibition in growth.The protein profile of stressed cells was compared with that of cellsgrown in the absence of nitrate using the iTRAQ peptide labellingstrategy and tandem liquid chromatography separation coupled with massspectrometry (quadrupoletime-of-flight) detection. A total of 737 uniqueproteins were identified by two or more peptides, representing 22 percentof the total DvH proteome and spanning every functional category. Theresults indicate that this was a mild stress, as proteins involved incentral metabolism and the sulphate reduction pathway were unperturbed.Proteins involved in the nitrate reduction pathway increased. Increasesseen in transport systems for proline, glycine^ betaineandglutamateindicate that the ...

Energy Technology Data Exchange (ETDEWEB)

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 efficiency for both ...

Energy Technology Data Exchange (ETDEWEB)

From 1977 through 1984, 293 previously untreated patients with biopsy proven carcinoma of the uterine cervix were treated by whole pelvis irradiation and high intensity 60Co remote afterloading (RAL) intrauterine tandem techniques in Haiti. The treatment results were analyzed retrospectively to evaluate the therapeutic results and prognostic factors of a strict protocol involving 40 Gy to the whole pelvis (2 Gy/day, 5 days/week). In addition, on the 5th day of the 3rd week, the first outpatient 60Co remote afterloading intracavitary insertion, delivering 7.5 Gy to point A with each insertion, repeated 3 times by a week separation for a total of 4 times. The total TDF for external beam plus RAL was 158 and 175 for early and late effects respectively. One hundred-four patients were evaluable after 1 year or more follow-up, with a median of 26.5 months. No evidence of disease (NED) by Stage at 1 year was: Stage I of 100% (3/3), Stage II of 82% (9/11), Stage III of 80% ...

Energy Technology Data Exchange (ETDEWEB)

N/C and S/C ratios show only minor differences between asphaltenes and coal on this basis, the asphaltenes continue to appear to be good surrogates for the characterization of organic acid rain precursors in coal. Alkylmethoxythiophene carboxylic acids (ATCA) are detected in all dichromate oxidation products analyzed to date. Relative concentrations of ATCA compounds are directly proportional to the sample`s bulk organic sulfur contents. Concentrations of ATCA compounds in oxidation products decrease upon repeated oxidation. This may indicate that the thiophenic groups tend to be located in external positions on the coal macromolecular structure and a high proportion of them can be stripped away with only one mild oxidation step. If true, this may make industrial-scale removal of organic sulfur easier than expected. Using analytical micropyrolysis-gas chromatography with a sulfur-selective flame photometric detector, it is possible to easily see a full distribution of organic sulfur ...

Energy Technology Data Exchange (ETDEWEB)

In the 14 MeV Neutron Laboratory, we have continued the development of a facility that is now the only one of its kind in operation in the United States. We have refined the klystron bunching system described in last year's report to the point that 1.2 nanosecond pulses have been directly measured. We have tested the pulse shape discrimination capability of our primary NE 213 neutron detector. We have converted the RF sweeper section of the beamline to a frequency of 1 MHz to replace the function of the high voltage pulser described in last year's report which proved to be difficult to maintain and unreliable in its operation. We have also overcome several other significant experimental difficulties, including a major problem with a vacuum leak in the main accelerator column. We have completed additional testing to prove the remainder of the generation and measurement systems, but overcoming some of these experimental difficulties has delayed the start of actual data ...

International Nuclear Information System (INIS)

Full text of publication follows: Understanding the dynamic irradiation effects on silica glasses is important for developing the diagnostic systems used in fusion and fission environments. While fundamental defects having an un-pared electron such as the E' center have been extensively studied, the neutral oxygen deficiency defects have been insufficiently clarified for lack of the detection methods. The ion induced luminescence is one of the probes that can be used to detect non-paramagnetic defects, and to observe creation and annihilation behavior dynamically. In the present study, we examined the characteristics of the ion induced luminescence such as energy, fluence flux and temperature dependence of the luminescence efficiency to analyze damage process quantitatively. Samples of SiO2 glasses were commercially available fused and synthesized silica glasses, produced by Toshiba Ceramics, Co. Ltd.. A thin films of SiO2 deposited on a Si wafer was used to determine the luminescence ...

International Nuclear Information System (INIS)

local peanut cultivars Giza 4 and Giza 5 were exposed to different doses ( 50,100,150,200,250 and 300 Gy)to Co "6"0 gamma ray source . all irradiated materials were cultivated to give the M_1 and M_2 generation , then high yielding mutants were selected from the M_2 generation and cultivated separately to give the M_3 generation. some yield component traits were measured for the three generations (M_1,M_2 and M_3) and M_3 families.variations observed in these traits were attributed to genetic factors. the M_3 selected mutants were artificially infected by aspergillus flavus L spores (aflatoxins-B1 and B2 groups-producer strain). aflatoxin concentrations were estimated and the analysis showed that aflatoxin concentrations in seeds of the M_3 selected mutants from cultivar Giza 4 were higher than those in seeds of the M3 mutants selected from cultivar Giza 5. however, the concentration of aflatoxin increased when the yield ( weight of seeds per plant trait) increased. ...

Science.gov (United States)

The rate of hepatic cytochrome P450 Cypla1 and Cyp1a2 induction was investigated in C57BL male mice during induction with o-aminoazotoluene (OAT), benzo[a]pyrene (BP) and 1,4-dihydroxyanthraquinone (AQ). The Cypla1 mPNA level determined by quantitative RT-competitive PCR increased more than three orders of magnitude during induction with OAT and BP compared with untreated animals and remained unchanged during induction with AQ. The Cypla2 mRNA level was only 8.5, 18.7 and 1.9 times higher during induction with OAT, BP and AQ respectively than in untreated mice. At the same time 7-Ethoxyresorufin-O-deethylase (EROD) and 7-Methoxyresorufin-O-demethylase (MROD) activities of Cypla were also investigated in liver. The increase of Cypla1 mRNA level correlated with the increase of EROD activity. This suggests involvement of the transcriptional mechanism of the inducibility of this enzyme. In the case of Cypla2 there was insignificant increase of its mRNA level but high ...

Science.gov (United States)

The GntR-like protein NorG has been shown to affect Staphylococcus aureus genes involved in the resistance to quinolones and ?-lactams, such as those encoding the NorB and AbcA transporters. To identify the target genes regulated by NorG, we carried out transcriptional profiling assays using S. aureus RN6390 and its isogenic norG::cat mutant. Our data showed that NorG positively affected the transcription of global regulators mgrA, arlS, and sarZ. The three putative drug efflux pump genes most positively affected by NorG were the NorB efflux pump (5.1-fold), the MmpL-like protein SACOL2566 (5.2-fold), and the BcrA-like drug transporter SACOL2525 (5.7-fold). The S. aureus predicted MmpL protein showed 53% homology with the MmpL lipid transporter of Mycobacterium tuberculosis, and the putative SACOL2525 protein showed 87% homology with the bacitracin drug transporter BcrA of Staphylococcus hominis. Two pump genes most negatively affected by NorG were NorC (4-fold) and AbcA (6-fold). ...

International Nuclear Information System (INIS)

Two candidate small interfering RNAs (siRNAs) corresponding to severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike gene were designed and in vitro transcribed to explore the possibility of silencing SARS-CoV S gene. The plasmid pEGFP-optS, which contains the codon-optimized SARS-CoV S gene and expresses spike-EGFP fusion protein (S-EGFP) as silencing target and expressing reporter, was transfected with siRNAs into HEK 293T cells. At various time points of posttransfection, the levels of S-EGFP expression and amounts of spike mRNA transcript were detected by fluorescence microscopy, flow cytometry, Western blot, and real-time quantitative PCR, respectively. The results showed that the cells transfected with pEGFP-optS expressed S-EGFP fusion protein at a higher level compared with those transfected with pEGFP-S, which contains wildtype SARS-CoV spike gene sequence. The green fluorescence, mean fluorescence intensity, and SARS-CoV S RNA ...

International Nuclear Information System (INIS)

High quality "3"1P MR spectra (signal to noise ratio (S/N) approximately 18, 15 min acquisition for each spectrum) were consistently obtained with surface coils over a period of 6-week RT. Both transient and steady state alterations in metabolites in response to RT were found in this case. The transient changes occurred during the first 3 hr immediately after the 3rd fractionated RT, these changes include the transient elevation of the PCr resonance, a decrease in PDE and an increase in intracellular pH. The monitoring showed that the metabolites approached steady state approximately 2 hr after the fractionated radiation intervention, suggesting that in vivo MRS can be useful for studying the dynamics of tumor response to RT such as repair of potential lethal damage, growth delay, and reoxygenation etc. The steady-state MR spectra showed the net response to each intervention and can clinically be useful for predicting and measuring the result of the fractionated ...

Science.gov (United States)

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

Science.gov (United States)

The complexity and diversity of the microbial communities in biogranules from an upflow anaerobic sludge blanket (UASB) bioreactor were determined in response to short-term changes in substrate feeds. The reactor was fed simulated brewery wastewater (SBWW) (70% ethanol, 15% acetate, 15% propionate) for 1.5 months (phase 1), acetate / sulfate for 2 months (phase 2), acetate-alone for 3 months (phase 3), and then a return to SBWW for 2 months (phase 4). Performance of the reactor remained relatively stable throughout the experiment as shown by COD removal and gas production. 16S rDNA, methanogen-associated mcrA and sulfate reducer-associated dsrAB genes were PCR amplified, then cloned and sequenced. Sequence analysis of 16S clone libraries showed a relatively simple community composed mainly of the methanogenic Archaea (Methanobacterium and Methanosaeta), members of the Green Non-Sulfur (Chloroflexi) group of Bacteria, followed by fewer numbers of Syntrophobacter, ...

International Nuclear Information System (INIS)

IL-4-induced Stat6 signaling is active in a variety of cell types and plays a role in cell proliferation/growth and resistance to apoptosis. Using EMSA, we identified differential IL-4/Stat6 activities in colorectal cancer cell lines, HT-29 being active Stat6"h"i"g"h phenotype and Caco-2 being defective Stat6"n"u"l"l phenotype, respectively. Active Stat6"h"i"g"h HT-29 cells exhibited resistance to apoptosis by flowcytometry and aggressive metastasis by Transwell assay compared with defective Stat6"n"u"l"l Caco-2 cells. Comparing one another using RT-PCR, Stat6"h"i"g"h HT-29 cells expressed more mRNA of anti-apoptotic and pro-metastatic genes Survivin, MDM2, and TMPRSS4, while Stat6"n"u"l"l Caco-2 cells expressed more mRNA of pro-apoptotic and anti-metastatic genes BAX, CAV1, and P53, respectively. This is the first study describing correlations of IL-4/Stat6 activities with apoptosis and metastasis in colon cancer. These findings, together with the observation of ...

International Nuclear Information System (INIS)

Isolation and expansion of neural stem cells (NSCs) of human origin are crucial for successful development of cell therapy approaches in neurodegenerative diseases. Different epigenetic and genetic immortalization strategies have been established for long-term maintenance and expansion of these cells in vitro. Here we report the generation of a new, clonal NSC (hc-NSC) line, derived from human fetal cortical tissue, based on v-myc immortalization. Using immunocytochemistry, we show that these cells retain the characteristics of NSCs after more than 50 passages. Under proliferation conditions, when supplemented with epidermal and basic fibroblast growth factors, the hc-NSCs expressed neural stem/progenitor cell markers like nestin, vimentin and Sox2. When growth factors were withdrawn, proliferation and expression of v-myc and telomerase were dramatically reduced, and the hc-NSCs differentiated into glia and neurons (mostly glutamatergic and GABAergic, as well as tyrosine ...

Energy Technology Data Exchange (ETDEWEB)

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 remained unchanged under all situations. ...

International Nuclear Information System (INIS)

Ewing's sarcoma is a small round-cell tumor typically arising in the bones, rarely in soft tissues, of children and adolescents. Ewing's sarcoma has retained the most unfavorable prognosis of all primary musculoskeletal tumors. Prior to the use of multi-drug chemotherapy, long-term survival was less than 10%. The development of multi-disciplinary therapy with chemotherapy, irradiation, and surgery has increased current long-term survival rates in most clinical centers to greater than 50%. In addition, the preferred method of tumor resection has changed; limb salvage has nearly replaced amputation of the affected limb. Limb salvage procedures can be performed in place of amputation without compromising patient survival rates. Recent studies have revealed that the pathognomonic translocations involving the EWS gene on chromosome 22 and an ETS-type gene, which is most commonly the Fli1 gene on chromosome 11, are implicated in more than 95% of Ewing's sarcomas, primitive neuroectodermal ...

International Nuclear Information System (INIS)

Since 1988, the Expertise group of Molecular and Cellular Biology (MCB) is an important partner in the development of the Micro-Ecological Life Support System Alternative (MELiSSA). The MELiSSA was designed to allow a small crew to survive on an Antarctic, lunar or Mars outpost, and is a joint research project currently fostered by the European Space Agency, ESA. The MELiSSA functions through a series of five interconnected compartments, of which four are microbial bioreactors and was engineered to degrade organic waste, regenerate the outpost's atmosphere and water, and provide the crew with an additional vegetarian diet. The bioreactor of the third compartment provides the edible cyanobacteria and plants of the fourth compartment with nitrate instead of ammonium as a source of nitrogen. The two bacteria responsible for the biological transformation of ammonium to nitrate (nitrification) are Nitrosomonas europaea and Nitrobacter winogradskyi. Since all MELiSSA-reactors are to be ...

International Nuclear Information System (INIS)

To evaluate the role of phytochelatins and metallothioneins in heavy metal tolerance of black mangrove Avicennia germinans, 3-month-old seedlings were exposed to cadmium or copper for 30 h, under hydroponic conditions. Degenerate Mt2 and PCS primers were synthesized based on amino acid and nucleotide alignment sequences reported for Mt2 and PCS in other plant species found in GenBank. Total RNA was isolated from A. germinans leaves and two partial fragments of metallothionein and phytochelatin synthase genes were isolated. Gene expression was evaluated with reverse transcripatase-polymerase chain reaction (RT-PCR) amplification technique. Temporal analysis showed that low Cd"2"+ and Cu"2"+ concentrations caused a slight (but not significant) increase in AvMt2 expression after a 16 h exposure time, while AvPCS expression showed a significant increase under the same conditions but only after 4 h. Results strongly suggest that the rapid increase in AvPCS expression ...

International Nuclear Information System (INIS)

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