An ultrasensitive laser-induced fluorescence detector was used with capillary electrophoresis for the study of 5-carboxy-tetramethylrhodamine. The raw signal from the detector provided roughly...Full Text Available
We propose a novel method for ultrasensitive isotope analysis that combines magnetic mass selection, resonant charge-exchange neutralization, and resonant laser ionizaion. Our method attains high isotopic abundance selectivity by means of continuous multistage separation of ions stored in a small ring. For the environmentally interesting case of /sup 90/Sr versus /sup 88/Sr we estimate that sensitivity better than 10/sup -15/ for a throughput of 10/sup 13/ atoms/sec and an efficiency (after the ion source) greater than 10% are readily achievable.
Summary of recent advances (abstract)Nucleic acid-based nanotechnology has always been perceived as novel, but has begun to move from theoretical demonstrations to practical applications....Full Text Available
Some of most used indicators in marine ecology are nucleic acid-derived indices. They can be divided by target levels in three groups: 1) at the organism level as ecophysiologic indicators, indicators...Full Text Available
An interactive system for computer analysis of nucleic acid and protein sequences has been developed for the Los Alamos DNA Sequence Database. It provides a convenient way to search or verify various...Full Text Available
We describe a non-isotopic, semi-automated method for large-scale multiplex analysis of nucleic acid sequences, using the cystic fibrosis transmembrane regulator (CFTR) gene as an example. Products...Full Text Available
A nucleic acid extraction system that can handle small numbers of specimens with a short test turnaround time and short hands-on time is desirable for emergent testing. We performed a comparative validation...Full Text Available
The NucliSens easyMAG and BioRot 9604 automated nucleic acid extraction systems were evaluated and compared with the manual QIAamp (Qiagen) extraction method for their abilities to extract nucleic acid...Full Text Available
Locked nucleic acid (LNA) is a nucleic acid analogue containing one or more LNA nucleotide monomers with a bicyclic furanose unit locked in an RNA mimicking sugar conformation. This conformational restriction results in unprecedented hybridization affinity towards complementary single stranded RNA and thus, makes LNA uniquely suited for mimicking RNA structures and sequence specific targeting of RNA in vitro or in vivo. The focus of this paper is on LNAantisense, LNA-modified siRNA (siLNA), and detection and analysis of microRNAs by LNA-modified oligonucleotide probes.
We present interactive computer programs for the analysis of nucleic acid sequences. In order to handle these programs, minimum computer experience is sufficient. The nucleotide sequence of the human...Full Text Available
A method of patterning n-type GaAs, InP, InGaAs, and InGaAsP by photoelectrochemical (PEC) etching in conjunction with a submicron focused ion beam (FIB) at low dose is described. The ion beam is used to produce damage in a desired pattern in the material. Subsequent PEC etching of the material reveals the ion induced features in relief. The procedure is highly sensitive, requiring a dose of only 5 x 10/sup 9/ ions/cm/sup 2/ for the differential etch to become apparent. The sensitivity allows rapid pattern generation in our FIB system.
Smooth muscle cell (SMC) proliferation and migration are key processes that occur in the pathogenesis of atherosclerosis and post-angioplasty restenosis. In the present study, we designed locked nucleic...Full Text Available
We developed prototype chemistry for nucleic acid hybridization on our bead-based diagnostics platform and we established an automatable bead handling protocol capable of 50 part-per-billion (ppb) sensitivity. We are working towards a platform capable of parallel, rapid (10 minute), raw sample testing for orthogonal (in this case nucleic acid and immunoassays) identification of biological (and other) threats in a single sensor microsystem. In this LDRD we developed the nucleic acid chemistry required for nucleic acid hybridization. Our goal is to place a non-cell associated RNA virus (Bovine Viral Diarrhea, BVD) on the beads for raw sample testing. This key pre-requisite to showing orthogonality (nucleic acid measurements can be performed in parallel with immunoassay measurements). Orthogonal detection dramatically reduces false positives. We chose BVD because our collaborators ...
A commercially available synthetic nucleic acid probe (SNAP) conjugated to alkaline phosphatase was compared with standard culture techniques for detecting Campylobacter species. The SNAP was able to...Full Text Available
Six related Staphylococcus phages spontaneously produced various abnormal head and tail structures: (i) giant capsids which were tailed and apparently contained nucleic acid; (ii) regular and irregular...Full Text Available
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
Nucleic acids are molecules of choice for both established and emerging nanoscale technologies. These technologies benefit from large functional densities of ‘DNA processing elements’...Full Text Available
The relative sensitivities of different protocols for detecting cytomegalovirus nucleic acid sequences in histological specimens, using a biotinylated cDNA probe, were assessed. Several commonly used...Full Text Available
There are many methods for introducing random mutations into nucleic acid sequences. Previously, we described a suite of programmes for estimating the completeness and diversity of randomized DNA libraries...Full Text Available
The uptake and incorporation of (/sup 3/H)adenine as a measure of total microbial (bacteria and unicellular algae) nucleic acid synthesis depends on the validity of several assumptions. A unique characteristic of the (/sup 3/H)adenine method is the ability to measure the specific radioactivity (nCi pmol/sup -1/) of the immediate precursor pool of the adenine incorporated into nucleic acids. This measurement permits correction for isotope dilution when the added radiotracer mixes with exogenous pools of structurally related compounds before uptake and with endogenous pools after transport into the cells. The immediate precursors to (/sup 3/H)adenine incorporation into cellular RNA and DNA are ATP and dATP. Since the intracellular dATP pool is in isotopic equilibrium with the ATP pool, one can determine the specific radioactivity of both precursor pools by measuring that of the intracellular ATP pool. If the specific activity of the precursor ...
Retinoic acid inducible gene I (RIG-I) is a pattern recognition receptor (PRR) responsible for detection of nucleic acids from pathogens in the cytoplasm of infected cells and induction of type I interferon...Full Text Available
Clinical applications of genetic therapies, including delivery of short, interfering RNAs (siRNAs) for RNA interference (RNAi), are limited due to the difficulty of delivering nucleic acids...Full Text Available
BackgroundPrimary hepatocytes are the best resource for in vitro studies directed at understanding hepatic processes at the cellular and molecular levels, necessary...Full Text Available
Three nucleic acid probes, two for autotrophic ammonia-oxidizing bacteria of the beta subdivision of the class Proteobacteria and one for alpha subdivision nitrite-oxidizing bacteria, were developed...Full Text Available
The Protein Data Bank contains the description of more than 45,000 three-dimensional protein and nucleic-acid structures today. Started to exist as the computer-readable depository of crystallographic...Full Text Available
BackgroundThe fast growing Protein Data Bank contains the three-dimensional description of more than 45000 protein- and nucleic-acid structures today. The large majority of the data...Full Text Available
The study by impedance and photoelectrochemical measurements of passive films formed on an AISI 304 type stainless steel shows that important parameters of the electronic structure of these films are modified under ..cap alpha.. irradiation, namely: width of the space charge region, donors concentration and diffusion length for minority carriers. The consequences of ..cap alpha.. irradiation on localized corrosion processes are discussed.
The study by impedance and photoelectrochemical measurements of passive films formed on an AISI 304 type stainless steel shows that important parameters of the electronic structure of these films are modified under #alpha# irradiation, namely: width of the space charge region, donors concentration and diffusion length for minority carriers. The consequences of #alpha# irradiation on localized corrosion processes are discussed.
Discovery of better biomarkers for diagnosis, prognosis, and therapy-response prediction is the most critical task of a scientific quest aimed at developing newly designed, tailor-made therapies for patients with cancer. Consequently, a proteome wide analysis, in addition to genomic studies, is an absolute requirement for a complete functional understanding of tumor biology. Ultra-sensitive, high-performance Fourier transform ion cyclotron resonance (FTICR) mass spectrometry (MS) currently holds an important role in fulfilling the demands of biomarker discovery. In this review, we describe the applicability of FTICR MS for breast cancer proteomics, particularly for the analysis of complex protein mixtures obtained from a limited number of cells typically available from clinical specimens.
The objectives of this study were to evaluate the performance of the NucliSens easyMAG platform for nucleic acid extraction from different clinical specimens compared to NucliSens miniMAG platform and manual QIAGEN extraction. The NucliSens easyMAG and the NucliSens miniMAG showed equal performance on 215 throat swabs since real-time nucleic acid sequence-based amplification scored the same samples positive for Mycoplasma pneumoniae (n=9) and Chlamydia pneumoniae (n=5) RNAs, although internal control RNA was slightly better detected with the NucliSens easyMAG (99.3% versus 96.8%). NucliSens easyMAG extracted nucleic acids more efficiently (higher recovery and/or fewer inhibitors) compared to QIAGEN extraction by showing, on average, lower Ct values in real-time LightCycler PCR, although 4 individual specimen out of 45 were found positive only with QIAGEN. For nine M. pneumoniae-positive throat swabs, the mean difference in ...
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.
Abstract In 46 febrile neutropenic patients who had undergone haematopoietic stem cell transplantation, the fluorescence in situ hybridisation using peptide nucleic acid probes (PNA FISH), Gram stain/acridine orange leukocyte cytospin (Gram/AOLC), and differential time to positivity (DTP) methods were performed for detection of catheter related bloodstream infections (CRBSIs). As compared with the DTP method (which detected 11 patients with CRBSI), the PNA FISH and the Gram/AOLC methods detected ten of 11 CRBSI patients, resulting in a sensitivity, specificity, negative predictive value and positive predictive value of 91%, 100%, 97% and 100%, respectively, for the PNA FISH method as well as for the Gram/AOLC method.
The feasibility of utilizing photoelectrochemical and electrolytical technologies to convert energy from the sun and wind into hydrogen was studied. In exploring opportunities to reduce the cost of hydrogen production through interaction with the electric utility grid, it was found that direct photoelectrochemical (PEC) conversion of sunlight has the economic potential to compete with direct photovoltaic/electrolysis, notwithstanding the significant stability and efficiency issues that are still awaiting solution. Interaction with the grid, while maximizing electrolizer use, makes a significant impact on the economics of producing hydrogen by photovoltaic/electrolysis, making wind-based systems also more economical. Electrolysis was found to be the optimal solution only with electricity from renewable sources or with less expensive non-peak electricity. On the other hand, the delivered cost of hydrogen was found to the lowest when electricity ...
This report discusses the development of transition metal oxide semiconductors for photoelectrolysis of water. More specifically, it involves preparation of TiO/sub 2/ films by sputtering and evaluating their physicochemical characteristics primarily as they relate to the behaviour of the films as photoanodes. Impedance, photoelectrochemical, and photoconduction properties of TiO/sub 2/ films sputtered in pure O/sub 2/ onto heated substrates have been determined as a function of O/sub 2/ pressure during sputtering, film thickness, Pt overcoating, and cathodic treatment. The capacitance data before cathodic treatment are of the form expected. The capacitance is essentially independent of potential, while for potentials increasingly cathodic of this value, the capacitance increases very rapidly. Cathodic treatment alters the impedance characteristics of the films but leads to either no detectible change in their photoelectrochemical properties or ...
Photocurrent measurements for stainless steel type 304 in the as-received condition after dc and AV (alternating voltage) passivation have confirmed that significant changes of the passive film properties occur due to AV passivation. The photocurrents were the highest for the sample treated by the AV passivation process (AVPP). The bandgap energy increased from about 2.8 to about 3.05 eV for this treatment. Very similar results have been obtained for pure chromium, which suggests that the AVPP produces a thicker passive film with a less defective nature due to a partial dehydration of the chromium oxide phase in the film.
Model phosphate-metal solvation complexes have been studied by ab-initio self-consistent-field techniques. The complexes studied include (RO)2PO2-(R = H or CH3) with Li+, Na+, K+, Be++, Mg++, Ca++,...Full Text Available
Eilatin-containing ruthenium complexes bind to a broad range of different nucleic acids including: calf thymus (CT) DNA, tRNAPhe, polymeric RNAs and DNAs, and viral RNAs including the HIV-1...Full Text Available
The metabolism of pentose-phosphate was investigated in Plasmodium falciparum-infected normal and glucose-6-phosphate dehydrogenase (G6PD)-deficient human red blood cells in vitro. 5'-Phosphoribosyl-1-pyrophosphate...Full Text Available
We have isolated a human thrombomodulin cDNA, and a human genomic clone containing the putative promoter domain, as well as the translated and untranslated regions of the endothelial cell receptor....Full Text Available
W have introduced template-synthesized metal phosphate nanoparticle labels for electrochemical immunoassay. Such use of an apoferritin template offers a simple and convenient route to prepare metallic nanoparticle labels for electrochemical immunoassays and avoid the complicated and time-consuming nanoparticle synthesis process (QD synthesis). Releasing metal ions from metal phosphate in an acetate buffer (pH 4.6) eliminates the harsh condition in the traditional metallic nanoparticle dissolution (e.g., strong acid dissolution of QDs and gold nanoparticles). This method is ultrasensitive and its DL is low to 77fM. The simultaneous detection of multiple protein targets is easily performed by using different metal phosphate nanoparticle labels (cadmium phosphate and lead phosphate). This approach can be extended to prepare multiple metal (such as zinc, lead, cadmium, copper, indium, gold, silver) phosphate nanoparticle labels or hybrid metal (bimetallic or ...
Photoelectrochemical current response of passive film was investigated for pure Cr and Fe-xCr (x = 8, 14, 18) alloys polarised potentiostatically in 0.1 kmol m"-"3 H_2SO_4 solution. Photoelectrochemical action spectrum could be separated into two or three constituents. These components were considered to be derived from Cr_2O_3 (E_g"o"p"t#propor to#3.6 eV) and Cr(OH)_3 (E_g"o"p"t#propor to#2.5 eV), and possibly CrO OH. The optical band gap, E_g"o"p"t, of each component was almost constant for various applied potentials, polarisation periods, and substrate materials. Flat band potential E_j_b at which the polarity of photocurrent changes from negative to positive with increasing potential was determined for each phase. E_j_b for Cr(OH)_3 on Cr and Fe-Cr alloys was about 250 mV_A_g_/_A_g_C_l. E_j_b for Cr_2O_3 was about 700 mV for Cr and about 500 mV for Fe-Cr alloys. E_j_b of Cr_2O_3 for Fe-Cr alloys slightly shifted in noble direction with ...
Fe-doped TiO2 hollow spheres (Fe-THs) were synthesized by sol?gel process using carbon spheres as templates. The prepared samples were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), UV?vis diffuse reflectance spectrum (DRS), N2 adsorption?desorption isotherms, Electron paramagnetic resonance (EPR) spectroscopy and Photoluminescence emission spectroscopy (PL). UV?vis spectra showed that Fe3+ doping could extend the absorption edge to the visible region. EPR spectra showed that Fe3+ was incorporated into the crystal lattice of TiO2, which could inhibit the recombination of photo-induced electron?hole pairs and improve the photocatalytic activity. The photocatalytic activities of the prepared samples were evaluated for the degradation of dye Reactive Brillia...
The effect of ion implantation on the aqueous corrosion resistance of Zr-4 in deaerated 1N H_2SO_4 was studied with the potentiokinetic technique. The Zr-4 alloy was bombarded with 5x10"1"4-2x10"1"6 Ar/cm"2 of 190 keV. It was found that the passive current density of Zr-4 decreases with increasing implantation dose. Photoelectrochemical results show that the ion implantation of Ar in Zr-4 raises the flatband potential of its passive film. AES was employed to analyze the surface of the passive film of Zr-4. The decrease in passive current density may be attributed to a thickening oxide layer on Zr-4 and a decrease in concentration of oxygen vacancies in its passive film. ((orig.)).
Moss-Schottky plots and photoelectrochemical measurements were made on films formed at different potentials on AISI 304 stainless steel in a borate/boric acid solution, pH 9.2. The results allowed the determination of the semiconductive properties and band structure of the films, which account for the existence of two kinds of films depending on the formation potential. For potentials below 0 V (SCE), the results point out for a film with an inverse spinel structure constituted by Cr-substituted magnetite with two donor levels. Above 0 V only one donor level is detected, which should be Fe{sup 2 +} on tetrahedral sites.
Moss-Schottky plots and photoelectrochemical measurements were made on films formed at different potentials on AISI 304 stainless steel in a borate/boric acid solution, pH 9.2. The results allowed the determination of the semiconductive properties and band structure of the films, which account for the existence of two kinds of films depending on the formation potential. For potentials below 0 V (SCE), the results point out for a film with an inverse spinel structure constituted by Cr-substituted magnetite with two donor levels. Above 0 V only one donor level is detected, which should be Fe"2 "+ on tetrahedral sites.
In a work studying the corrosion and tarnishing properties of a variety of copper alloys, the alloy Cu-A110-Sn5 was found to show an excellent corrosion resistance in neutral solutions, where copper and most conventional Cu alloys are covered by thick nonprotective surface layers. The passive films formed on this alloy were characterized with electrochemical and photoelectrochemical methods. The pH dependence of the passivation and of the photocurrent behavior of the Cu-A110-Sn5 alloy clearly indicates that the passivity of this alloy in neutral solutions is due to a formation of passive film enriched with aluminum oxide. At corrosion potential a strong increase in the corrosion resistance with time is due to a gradual enrichment of the surface with aluminum oxide. This can be seen in the photocurrent spectra which change from cooper-type to aluminum-type with time. At higher applied potentials the formation of an aluminum-type oxide film is strongly accelerated.
Indium sulfide thin films were deposited onto indium-tin-oxide coated glass substrate by electrochemical deposition from an aqueous solution containing In2 (SO4) 3 and Na2S2O3. The deposition conditions were optimized on the basis of data obtained by scanning electron microscope, Auger electron spectroscopy and optical transmission measurements. Furthermore, the photosensitivity of the films was observed by means of photoelectrochemical measurements, which confirmed that the indium sulfide showed n-type conduction. The X-ray diffraction and Raman studies revealed that the as-grown films were amorphous or nanocrystalline in nature and became polycrystalline In2S3 after annealing.
The effect of Mo addition as an alloying element to stainless steel alloys is investigated by capacitance (Mott Schottky approach), and photoelectrochemistry measurements. Complementary studies were made using Auger electron spectroscopy and X-ray photoelectron spectroscopy. The Mott-Schottky approach and the photoelectrochemical studies showed that the presence of Mo as an alloying element affects the semiconductive properties of the oxide films. The analytical results have shown that the oxide films formed on stainless steels are composed by an external Fe rich region and an inner Cr rich region. No significant amount of Mo was found in the outer layers of the film. The presence of Mo leads to an increase of the chromium content in the inner layers of the film, although without increasing the film thickness. (orig.) 30 refs.
The effect of Mo addition as an alloying element to stainless steel alloys is investigated by capacitance (Mott Schottky approach), and photoelectrochemistry measurements. Complementary studies were made using Auger electron spectroscopy and X-ray photoelectron spectroscopy. The Mott-Schottky approach and the photoelectrochemical studies showed that the presence of Mo as an alloying element affects the semiconductive properties of the oxide films. The analytical results have shown that the oxide films formed on stainless steels are composed by an external Fe rich region and an inner Cr rich region. No significant amount of Mo was found in the outer layers of the film. The presence of Mo leads to an increase of the chromium content in the inner layers of the film, although without increasing the film thickness. (orig.)
The effect of UV radiation in the wavelength region 230 nm to 302 nm on the ability of an irradiated mammalian cell to reactivate UV-irradiated mammalian virus was tested. An action spectrum for radiation enhanced reactivation (RER) is presented. The shape of the action spectrum points to a combined nucleic acid-protein target for UV radiation effects on this cellular parameter. An analysis of the results of others involving the biochemical and photobiological events involved in RER does not allow us to distinguish which macromolecule is the major contributor to this effect. Studies involving an analogous phenomenon in bacteris (Weigle reactivation) imply that RER and WR amy involve similar mechanisms. (author).
Nucleic acid extraction and human immunodeficiency virus type 1 (HIV-1) genotyping using the NucliSens miniMAG platform and the TruGene HIV-1 genotyping kit gave HIV-1 sequence data from HIV-1-negative plasma spiked with 100 copies/ml reference HIV-1 RNA and from low-viremia clinical samples (<500 copies/ml) without the need for ultracentrifugation or nested second-round PCR. PMID:17122019
The term "homology" or "homologous" means an amino acid similarity measured by the program, BLAST (Altschul et al (1997), "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs", Nucleic Acids Res. 25:33 89 3402), and expressed as --(% identity n/n). In measuring homology between a peptide and a protein of greater size, homology is measured only in the corresponding region; that is, the protein is regarded as only having the same general length as the peptide, allowing for gaps and insertions.
The use of platinum(II) complexes as tags and probes for biomolecules is indeed advantageous for their reactivities can be selective for certain purposes through an interplay of mild reaction conditions and of the ligands bound to the platinum. The use of {sup 195}Pt NMR as a method of detecting platinum and its interactions with biomolecules was carried out with the simplest model of platinum(II) tagging to proteins. Variable-temperature {sup 195}Pt NMR spectroscopy proved useful in studying the stereodynamics of complex thioethers like methionine. The complex, Pt(trpy)Cl{sup +}, with its chromophore has a greater potential for probing proteins. It is a noninvasive and selective tag for histidine and cysteine residues on the surface of cytochrome c at pH 5. The protein derivatives obtained are separable, and the tags are easily quantitated and differentiated through the metal-to-ligand charge transfer bands which are sensitive to the environment of the tag. Increasing the pH to 7.0 ...
Photoelectrochemical oxidation is a potentially interesting method for destroying toxic organic materials. We have studied the photoelectrocatalytic activity of TiO{sub 2} films made by thermal oxidation of titanium, low pressure chemical vapour deposition (LPCVD), and anodisation of titanium. Two model organic compounds have been investigated for photooxidation: methyl phosphonic acid (MPA) which is a nerve gas analogue and 4-chlorophenol (4-CP) which is a chlorinated aromatic compound considered a standard for the evaluation of the TiO{sub 2} / UV processes. In addition to photoelectrochemical characterisation the films have been characterised by profilometry, XRD, AFM, photocurrent spectroscopy and Raman microscopy. Correlations have been made between the physical properties of the thin films and their catalytic activities. The most catalytic sample of thermally oxidised titanium was prepared at 400 deg C, and the presence of anatase was ...
Studies are made in this program to enhance our understanding of the role of 5f electrons in chemical bonding. Most of our studies on the macroscopic scale emphasize certain isotopes available from the National Transplutonium Production Program centered at HFIR/TRU -namely, _9_6"2"4"8Cm, _9_7"2"4"9Bk, _9_8"2"4"9Cf, _9_9"2"5"3Es, and _9_9"2"5"4Es. Particular classes of compounds, such as hydrides, chalcogenides, and pnictides are often selected for study on the basis of their utility in bringing out significant chemical differences between the lanthanides and the actinides. High temperature/high pressure experiments on actinide metals and compounds reveal characteristics about the degree of 5f electron localization as a function of atomic number in the series. Magnetic properties of metals and compounds are investigated using ultra-sensitive methods and hydration numbers of ions in solution are investigated by neutron scattering at HFIR. As a result of a recently ...
To accomplish the environmental radiological surveillance need of India's ambitious nuclear power programme, Health Physics Division is infusing new technologies and improved analytical techniques for day to day measurements of various radionuclides in different environmental matrices. It is essential to have techniques for measuring the concentration of radionuclides just above the background level since the discharges from the nuclear facilities are very low i.e. in the range of 5-10% of the prescribed discharge limits by the regulatory bodies. In view of developing ultra-sensitive techniques, the aim of ongoing programmes of the division is to meet the challenges of measuring ultra trace level of radioactivity by adopting state of art new instrumentation and improved sample processing techniques. This will allow us to measure the lowest level of radioactivity (3H, 90Sr, 137Cs, 239+240Pu, etc.) in the environment and thereby estimating the lowest doses to the ...
Abstract Hydrogen bonding interactions between amino acids and nucleic acid bases constitute the most important interactions responsible for the specificity of protein binding. In this study, complexes formed by hydrogen bonding interactions between cysteine and thymine have been studied by density functional theory. The relevant geometries, energies, and IR characteristics of hydrogen bonds (H-bonds) have been systematically investigated. The quantum theory of atoms in molecule and natural bond orbital analysis have also been applied to understand the nature of the hydrogen bonding interactions in complexes. More than 10 kinds of H-bonds including intra- and intermolecular H-bonds have been found in complexes. Most of intermolecular H-bonds involve O (or N) atom as H-acceptor, whereas the...
The possible long term effects of chronic radiation exposure have been studied. Rats were given tritiated drinking water (3#mu#Ci/ml) before pregnancy, during pregnancy and threafter, continuously through 5 generations. The brains were studied at 30 and 120 days postnatally. Significant decreases in weight, DNA and protein contents of specific parts of the brain were found, the most pronounced decreases being in the diencephalon. The decreases in protein content were more pronounced than the decreases in DNA content; the decreases were also more pronounced at 30 days than at 120 days, and more pronounced in the fifth generation than in previous generations. An attempt was made to explain these complex phenomena on the basis of known periods of proliferation of various brain cells, damage to mothers themselves in each generation, and damage and repair of nucleic acids caused by radioactivity.
The stages of processes leading to radiation damage are studied, as well as, the direct and indirect mechanics of its production. The radiation effects on nucleic acid and protein macro moleculas are treated. The physical and chemical factors that modify radiosensibility are analysed, in particular the oxygen effects, the sensibilization by analogues of nitrogen bases, post-effects, chemical protection and inherent cell factors. Consideration is given to restoration processes by excision of injured fragments, the bloching of the excision restoration processes, the restoration of lesions caused by ionizing radiations and to the restoration by genetic recombination. Referring to somatic effects of radiation, the early ones and the acute syndrome of radiation are discussed. The difference of radiosensibility observed in mammalian cells and main observable alterations in tissues and organs are commented. Referring to delayed radiation effects, carcinogeneses, ...
Semiconductor properties of passive films formed on the Fe-18Cr alloy in a borate buffer solution (pH = 8.4) and 0.1 M H{sub 2}SO{sub 4} solution were examined using a photoelectrochemical spectroscopy and an electrochemical impedance spectroscopy. Photo current reveals two photo action spectra that derived from outer hydroxide and inner oxide layers. A typical n-type semiconductor behaviour is observed by both photo current and impedance for the passive films formed in the borate buffer solution. On the other hand, a negative photo current generated, the absolute value of which decreased as applied potential increased in the sulfuric acid solution. This indicates that the passive film behaves as a p-type semiconductor. However, Mott-Schottky plot revealed the typical n-type semiconductor property. It is concluded that the passive film on the Fe-18Cr alloy formed in the borate buffer solution is composed of both n-type outer hydroxide and inner oxide layers. On the ...
Semiconductor properties of passive films formed on the Fe-18Cr alloy in a borate buffer solution (pH = 8.4) and 0.1 M H_2SO_4 solution were examined using a photoelectrochemical spectroscopy and an electrochemical impedance spectroscopy. Photo current reveals two photo action spectra that derived from outer hydroxide and inner oxide layers. A typical n-type semiconductor behaviour is observed by both photo current and impedance for the passive films formed in the borate buffer solution. On the other hand, a negative photo current generated, the absolute value of which decreased as applied potential increased in the sulfuric acid solution. This indicates that the passive film behaves as a p-type semiconductor. However, Mott-Schottky plot revealed the typical n-type semiconductor property. It is concluded that the passive film on the Fe-18Cr alloy formed in the borate buffer solution is composed of both n-type outer hydroxide and inner oxide layers. On the other ...
A theoretical model, which describes subband gap photoexcitation involving localized electronic states, was developed. The escape probability of a charge carrier trapped in a localized state is considered via Poole-Frenkel, direct tunneling, or phonon-assisted tunneling processes, as competing escape mechanisms. Photoelectrochemical experiments were performed on the passive films formed on zirconium and amorphous iron-zirconium alloys and on pure HfO/sub 2/ films and HfO/sub 2/ films implanted with varying concentrations of xenon. These films were found to possess some degree of disorder depending on the substrate, the thickness of the film, and the extent of implantation. The spectral dependence of the photocurrent in all of the films studied is considerably different from what was found for crystalline passive films. The potential dependence of the photocurrent yields Poole-Frenkel behavior. Reverse tunneling processes were also observed at low photon energies ...
Electronic properties of passive films formed on Fe at various applied potentials in pH 8.5 buffer solution were examined through the photocurrent measurement and impedance spectroscopy. Passive film formed on Fe at relatively low potentials was found to be r-FeOOH layer and internal r-Fe{sub 2}O{sub 3} layer. However, the r-FeOOH layer became unstable and disappeared at potentials below 400mV and hence may be an adsorbed layer. An electronic band structure model for the passive film of Fe was suggested on the basis of the spinel band model with involving two types of electronic excitation processes, i. e., the p-d and the d-d transition together. The effects of applied potential on the photocurrent behaviors of the passive film on Fe were explained appropriately by separating the photocurrent spectra depending on the transition type. The Mott-Schottky behavior for the passive film on Fe and the photocurrent variation with applied potential were also analyzed in terms of the suggested ...
The electrochemical method has been used to deposit zinc oxide (ZnO) thin films from aqueous zinc nitrate solution at 80 deg. C onto fluorine doped tin oxide (FTO) coated glass substrates. ZnO thin films were grown between - 0.900 and - 1.025 V vs Ag/AgCl as established by voltammogram. Characterisation of ZnO films was carried out for both as-deposited and annealed films in order to study the effect of annealing. Structural analysis of the ZnO films was performed using X-ray diffraction, which showed polycrystalline films of hexagonal phase with (002) preferential orientation. Atomic force microscopy was used to study the surface morphology. Optical studies identified the bandgap to be {approx} 3.20 eV and refractive index to 2.35. The photoelectrochemical cell signal indicated that the films had n-type electrical conductivity and current-voltage measurements showed the glass/FTO/ZnO/Au devices exhibit rectifying properties. The thickness of the ZnO films was ...
The effect of molybdenum on the electronic structure of the passive films formed on ferritic (Fe-Cr and Fe-Cr-Mo) stainless steels is examined by capacitance and photoelectrochemical measurements. The capacitance study is supported by a mathematical analysis of the Schottky barrier developed at the semiconductor-electrolyte interface in the case of a semiconductor with multiple bulk electronic states in the bandgap. The numerical simulations, based on the more general Mott-Schottky relation proposed, are in good agreement with the experimental results. It can be concluded that the capacitance behavior of the passive films is related to the contributions of a shallow donor level very close to the conduction band and a deep donor level at about 0.4 eV below the conduction band. The addition of molybdenum decreases the donor density of the deep level. Photoeffects observed for subbandgap photon energies reveal that this deep donor level behaves like a trapping ...
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 ...
Rapid detection and characterization of food borne pathogens such as Escherichia coli O157:H7 is crucial for epidemiological investigations and food safety surveillance. As an alternative to conventional technologies, we examined the sensitivity and specificity of nucleic acid microarrays for detecting and genotyping E. coli O157:H7. The array was composed of oligonucleotide probes (25-30 mer) complementary to four virulence loci (intimin, Shiga-like toxins I and II, and hemolysin A). Target DNA was amplified from whole cells or from purified DNA via single or multiplexed polymerase chain reaction (PCR), and PCR products were hybridized to the array without further modification or purification. The array was 32-fold more sensitive than gel electrophoresis and capable of detecting amplification products from < 1 cell equivalent of genomic DNA (1 fg). Immunomagnetic capture, PCR and a microarray were subsequently used to detect 55 CFU ml-1 (E. coli O157:H7) ...
The feasibility of containing genetically engineered bacteria with enhanced dehalogenating properties for in situ bioremediation was investigated. (1) An agarose matrix microbead protocol and a detection system for contained microorganisms or DNA were developed. Multiplex Polymerase Chain Reaction (PCR) allowed tracking of a consortium of encapsulated organisms or several gene targets from a single species. Gene sequences encoding the enzymes responsible for initiating the biodegradation of toluene, octane, and 2,4-D were detected by multiplex PCR and nucleic acid probes from similar to 1-10 biodegradative cells/g soil. Improved DNA extraction methods resulted in PCR reactions detecting similar to 6 cells/g soil. (2) The pcpB gene (for the broad-spectrum detoxicant pentachlorophenol (PCP) hydroxylase) isolated from Flavobacterium sp. strain ATCC 39723 was used in attempts to develop an improved dehalogenating recombinant microorganism for containment experiments.
Modification of DNA and double-stranded deoxyoligonucleotides with antitumor 1,2-diamino-cyclohexanedinitroplatinum(II) (Pt-dach) complexes was investigated with the aid of physico-chemical methods and chemical probes of nucleic acid conformation. The three Pt-dach complexes were used which differed in isomeric forms of the dach non-leaving ligand-Pt(1R,2R-dach), Pt(1S,2S-dach) and Pt(1R,2S-dach) complexes. The latter complex has lower antitumor activity than the other two Pt-dach complexes. Pt(1R,2S-dach) complex exhibits the slowest kinetics of its binding to DNA and of the conversion of monofunctional binding to bifunctional lesions. The anomalously slow electrophoretic mobility of multimers of the platinated and ligated oligomers suggests that bifunctional binding of Pt-dach complexes to a d(GG) site within double-stranded oligonucleotides induces bending of the oligomer. It is proposed that different effects of the Pt-dach complexes on DNA observed in this ...
Pyrrolo[2,1-c][1,4]benzodiazepine (PBD) chemicals are antitumor antibiotics inhibiting nucleic acid synthesis. An indole carboxylate-PBD hybrid with six-carbon spacer structure (IN6CPBD) has been previously demonstrated to induce melanoma cell apoptosis and reduce metastasis in mouse lungs. This study aimed at investigating the efficacy of the other hybrid compound with four-carbon spacer (IN4CPBD) and elucidating its anti-metastatic mechanism. Human melanoma A375 cells with IN4CPBD treatment underwent cytotoxicity and apoptosis-associated assays. Transwell migration assay, Western blotting, and ELISA were used for mechanistic study. IN4CPBD exhibited potent melanoma cytotoxicity through interrupting G1/S cell cycle progression, increasing DNA fragmentation and hypodipoidic DNA contents, a...
A combined application of several microtechniques is presented and discussed with the Ti/TiO_2 and Zr/ZrO_2-systems as an example. All measurements were carried out on single grains of technical materials in order to detect and quantify the effect of substrate microstructure on the properties of anodic passive films formed potentiodynamically in 0.5 M H_2SO_4 (dU/dt = 20 mVs"-"1). Anisotropy-micro-ellipsometry (AME) was employed to determine the crystallographic orientation of the substrate grains along with passive film thickness and crystallinity in dependence on the anodization potential. Both the isotropic (amorphous) TiO_2- and the anisotropic (crystalline) ZrO_2-films exhibit a systematic dependence of film thickness on the grain orientation. Local LASER-scanning photocurrent measurements (#lambda#=257 nm) on the same grains likewise show a heterogeneity of the photoelectrochemical reactivity in all cases. This is quantitatively explained by the results from ...
PCR detection of genetic material of the parasites present in faeces may be an alternative for microscopic and serological tests routinely used for diagnosing parasitic enteral infections. However, small amount of target DNA combined with low efficiency of total DNA extraction, and presence of PCR inhibitors in the samples to be amplified, may cause false negative detection results. The aim of this work was to evaluate the impact of DNA isolation procedure used on the amplification of DNA fragments from the genomes of protozoan Cryptosporidium parvum and the nematode Trichinella spiralis. Two methods based on different principles of biological material lysis were evaluated; NucliSENS miniMAG employing simultaneously applied chemical lysis and mechanical disruption or mechanical disruption followed by enzymatic lysis in case of QIAamp DNA Stool Mini Kit. Both of the analyzed systems for nucleic acids purification allowed isolation of DNA from purified ...
Clostridium difficile (C. difficile) causes 25-30% of cases of antibiotic associated diarrhea and most cases of pseudomembranous colitis. Patients presenting with diarrhea after hospitalization for 3 or more days should be tested for C. difficile. There are many options available for testing, each of which has inherent advantages and disadvantages. Most laboratories perform toxin testing using an enzyme immunoassay method. In general these tests have sensitivities ranging from 60 to 70% and specificities of 98%. When using these methods, symptomatic patients with negative tests should be tested by another more sensitive method. Until recently, cell culture cytotoxicity neutralization assays (CCNAs) were considered the gold standard in the U.S. A two-step algorithm using an EIA for glutamate dehydrogenase detection followed by testing positives using CCNA, offered an improved alternative until the availability of molecular assays. Although early studies that compared the GDH assay to ...
In this paper author deals with the regulation of electricity prices in the Slovak Republic. Author contests the social policy of the government through doped prices of electricity. Two thirds of electricity is generated in nuclear power plants in Slovakia. Hence, it is necessary to focus on the solution of problem of nuclear waste. In 2004 Ministry of Economy stated, that the deficit in nuclear fund, from which the country have to fully cover the costs of liquidation and final disposal of nuclear waste, is estimated in the amount of around 89 billion Slovak crowns (? 3.7 billion $). From it, so called historical deficit, which originated because of late foundation of fund, represents officially 15 billion Slovak crowns (? 0.62 billion $). In Slovakia exists the real risk, that by maintenance of present state by creation and draw of the fund, it will be possible to ensure only 39 per cent of financial sources necessary for full financial handling of the back part of nuclear energetic. ...
The thermostable Thermus aquaticus DNA polymerase (Taq Pol) has been the key factor in transforming the initial PCR method into one with huge impact in molecular biology and biotechnology. Therefore, the development of effective affinity adsorbents for the purification of Taq Pol, as well as other DNA polymerases, attracts the attention of the enzyme manufacturers and the research laboratories. In this report we describe a simple protocol for the purification of Taq Pol from E. coli lysates, leading to enzymes of high specific activity and purity. The protocol is based on a single affinity chromatography step, featuring an immobilized ligand selected from a structure-biased combinatorial library of dNTP-mimetic synthetic ligands. The ligand library was screened for its ability to bind and purify Taq Pol from E. coli lysates. One immobilized ligand (mABSGu) of the general formula X-Trz-Y, bearing 9-aminoethylguanine (AEGu) and aniline-2-sulfonic acid (mABS) linked on the triazine ...
There are many methods for introducing random mutations into nucleic acid sequences. Previously, we described a suite of programmes for estimating the completeness and diversity of randomized DNA libraries generated by a number of these protocols. Our programmes suggested some empirical guidelines for library design; however, no information was provided regarding library diversity at the protein (rather than DNA) level. We have now updated our web server, enabling analysis of translated libraries constructed by site-saturation mutagenesis and error-prone PCR (epPCR). We introduce GLUE-Including Translation (GLUE-IT), which finds the expected amino acid completeness of libraries in which up to six codons have been independently varied (according to any user-specified randomization scheme). We provide two tools for assisting with experimental design: CodonCalculator, for assessing amino acids corresponding to given randomized codons; and AA-Calculator, for finding ...
Owing to exceptional biomolecule preservation, fossil avian eggshell has been used extensively in geochronology and palaeodietary studies. Here, we show, to our knowledge, for the first time that fossil eggshell is a previously unrecognized source of ancient DNA (aDNA). We describe the successful isolation and amplification of DNA from fossil eggshell up to 19 ka old. aDNA was successfully characterized from eggshell obtained from New Zealand (extinct moa and ducks), Madagascar (extinct elephant birds) and Australia (emu and owl). Our data demonstrate excellent preservation of the nucleic acids, evidenced by retrieval of both mitochondrial and nuclear DNA from many of the samples. Using confocal microscopy and quantitative PCR, this study critically evaluates approaches to maximize DNA recovery from powdered eggshell. Our quantitative PCR experiments also demonstrate that moa eggshell has approximately 125 times lower bacterial load than bone, making it a highly ...
In view of the anticipated increased use of atomic energy in industry, the possible long-term effects of chronic radiation exposure were studied in five generations of rats. Female rats (F_0) were given tritiated drinking water ("3HOH;3#mu#Ci/ml) from adolescence (60 days) until and throughout pregnancy. A separate study showed that the maximum radioactivity in the urine is reached in 30 days, and in the blood in 42 days. In the newborns, the highest specific activity was in the nucleic acid fraction, but total radioactivity was mainly due to the water (body fluids) compartment. No signs of radiation illness or increase in cataract formation in the mothers were observed. The food and water intake and body weight changes before pregnancy were normal. The course and the outcome of pregnancy were also normal. However, 60% of the newborns (F_1) exhibited hematomas, edemas, and subdural hemorrhages, which disappeared at 30 days of age. Bollod analysis of the F_1 ...
An attempt was made for early quantitative assessment of the severety of an injury, using the nuclear acids (NA) content as a biochemical indicator. Sexually mature male rats with body mass 160-180 g were irradiated with doses 0.5, 1, 2, 4 and 8 Gy of "1"3"7Cs source. Uneven irradiation was achieved by shielding the abdominal-lumbar area with lead rings, measuring 2,5/50 mm. A 28-30% fall of the dose beneath the shielded zone was thus provided. Dose-dependent quantitative changes in Na content were determined on hours 24 and 72 within the range from 0.5 to 8 Gy and followed up until day 30 after irradiation with nonlethal (2 Gy) and lethal (8 Gy) doses. The total NA content in the leucocytes was determined by the method of Kritski and Aleksandrov. The uneven irradiation of rats within the 0,5-8 Gy range induced early dose-dependent changes in NA content. The nature of their post-irradiation dynamics also depended on the radiation dose and adequately reflected the periods of development ...
The collection of papers in this issue of Mycologia documents considerable improvements in taxon sampling and phylogenetic resolution regarding the Fungal Tree of Life. The new data will stimulate new attempts to date divergences and correlate events in fungal evolution with those of other organisms. Here, we review the history of dating fungal divergences by nucleic acid variation and then use a dataset of 50 genes for 25 selected fungi, plants and animals to investigate divergence times in kingdom Fungi. In particular, we test the choice of fossil calibration points on dating divergences in fungi. At the scale of our analysis, substitution rates varied without showing significant within-lineage correlation, so we used the Langley-Fitch method in the R8S package of computer programs to estimate node ages. Different calibration points had a dramatic effect on estimated divergence dates. The estimate for the age of the Ascomycota/Basidiomycota split was 1808000000 y ...
We report a search for small RNAs (sRNAs) in the low-GC, gram-positive human pathogen Streptococcus pneumoniae. Based on bioinformatic analyses by Livny et al. (J. Livny, A. Brencic, S. Lory, and M. K. Waldor, Nucleic Acids Res. 34:3484-3493, 2006), we tested 40 candidates by Northern blotting and confirmed the expression of nine new and one previously reported (CcnA) sRNAs in strain D39. CcnA is one of five redundant sRNAs reported by Halfmann et al. (A. Halfmann, M. Kovacs, R. Hakenbeck, and R. Bruckner, Mol. Microbiol. 66:110-126, 2007) that are positively controlled by the CiaR response regulator. We characterized 3 of these 14 sRNAs: Spd-sr17 (144 nucleotides [nt]; decreased in stationary phase), Spd-sr37 (80 nt; strongly expressed in all growth phases), and CcnA (93 nt; induced by competence stimulatory peptide). Spd-sr17 and CcnA likely fold into structures containing single-stranded regions between hairpin structures, whereas Spd-sr37 forms a base-paired ...