Interstrand cross-links arising from strand breaks at true abasic sites in duplex DNA.

Science.gov (United States)

Yang, Zhiyu; Price, Nathan E; Johnson, Kevin M; Wang, Yinsheng; Gates, Kent S

2017-06-20

Interstrand cross-links are exceptionally bioactive DNA lesions. Endogenous generation of interstrand cross-links in genomic DNA may contribute to aging, neurodegeneration, and cancer. Abasic (Ap) sites are common lesions in genomic DNA that readily undergo spontaneous and amine-catalyzed strand cleavage reactions that generate a 2,3-didehydro-2,3-dideoxyribose sugar remnant (3'ddR5p) at the 3'-terminus of the strand break. Interestingly, this strand scission process leaves an electrophilic α,β-unsaturated aldehyde residue embedded within the resulting nicked duplex. Here we present evidence that 3'ddR5p derivatives generated by spermine-catalyzed strand cleavage at Ap sites in duplex DNA can react with adenine residues on the opposing strand to generate a complex lesion consisting of an interstrand cross-link adjacent to a strand break. The cross-link blocks DNA replication by ϕ29 DNA polymerase, a highly processive polymerase enzyme that couples synthesis with strand displacement. This suggests that 3'ddR5p-derived cross-links have the potential to block critical cellular DNA transactions that require strand separation. LC-MS/MS methods developed herein provide powerful tools for studying the occurrence and properties of these cross-links in biochemical and biological systems. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

Correlation of bistranded clustered abasic DNA lesion processing with structural and dynamic DNA helix distortion

Science.gov (United States)

Bignon, Emmanuelle; Gattuso, Hugo; Morell, Christophe; Dehez, François; Georgakilas, Alexandros G.; Monari, Antonio; Dumont, Elise

2016-01-01

Clustered apurinic/apyrimidinic (AP; abasic) DNA lesions produced by ionizing radiation are by far more cytotoxic than isolated AP lesion entities. The structure and dynamics of a series of seven 23-bp oligonucleotides featuring simple bistranded clustered damage sites, comprising of two AP sites, zero, one, three or five bases 3′ or 5′ apart from each other, were investigated through 400 ns explicit solvent molecular dynamics simulations. They provide representative structures of synthetically engineered multiply damage sites-containing oligonucleotides whose repair was investigated experimentally (Nucl. Acids Res. 2004, 32:5609-5620; Nucl. Acids Res. 2002, 30: 2800–2808). The inspection of extrahelical positioning of the AP sites, bulge and non Watson–Crick hydrogen bonding corroborates the experimental measurements of repair efficiencies by bacterial or human AP endonucleases Nfo and APE1, respectively. This study provides unprecedented knowledge into the structure and dynamics of clustered abasic DNA lesions, notably rationalizing the non-symmetry with respect to 3′ to 5′ position. In addition, it provides strong mechanistic insights and basis for future studies on the effects of clustered DNA damage on the recognition and processing of these lesions by bacterial or human DNA repair enzymes specialized in the processing of such lesions. PMID:27587587

Clustered DNA damages induced in isolated DNA and in human cells by low doses of ionizing radiation

Science.gov (United States)

Sutherland, B. M.; Bennett, P. V.; Sidorkina, O.; Laval, J.; Lowenstein, D. I. (Principal Investigator)

2000-01-01

Clustered DNA damages-two or more closely spaced damages (strand breaks, abasic sites, or oxidized bases) on opposing strands-are suspects as critical lesions producing lethal and mutagenic effects of ionizing radiation. However, as a result of the lack of methods for measuring damage clusters induced by ionizing radiation in genomic DNA, neither the frequencies of their production by physiological doses of radiation, nor their repairability, nor their biological effects are known. On the basis of methods that we developed for quantitating damages in large DNAs, we have devised and validated a way of measuring ionizing radiation-induced clustered lesions in genomic DNA, including DNA from human cells. DNA is treated with an endonuclease that induces a single-strand cleavage at an oxidized base or abasic site. If there are two closely spaced damages on opposing strands, such cleavage will reduce the size of the DNA on a nondenaturing gel. We show that ionizing radiation does induce clustered DNA damages containing abasic sites, oxidized purines, or oxidized pyrimidines. Further, the frequency of each of these cluster classes is comparable to that of frank double-strand breaks; among all complex damages induced by ionizing radiation, double-strand breaks are only about 20%, with other clustered damage constituting some 80%. We also show that even low doses (0.1-1 Gy) of high linear energy transfer ionizing radiation induce clustered damages in human cells.

Repair of endogenous and ionizing radiation-induced DNA damages: mechanisms and biological functions

International Nuclear Information System (INIS)

Boiteux, S.

2002-01-01

The cellular DNA is continuously exposed to endogenous and exogenous stress. Oxidative stress due to cellular metabolism is the major cause of endogenous DNA damage. On the other hand, ionizing radiation (IR) is an important exogenous stress. Both induce similar DNA damages: damaged bases, abasic sites and strand breakage. Most of these lesions are lethal and/or mutagenic. The survival of the cell is managed by efficient and accurate DNA repair mechanisms that remove lesions before their replication or transcription. DNA repair pathways involved in the removal of IR-induced lesions are briefly described. Base excision repair (BER) is mostly involved in the removal of base damage, abasic sites and single strand breaks. In contrast, DNA double strand breaks are mostly repaired by non-homologous end joining (NHEJ) or homologous recombination (HR). How DNA repair pathways prevent cancer process is also discussed. (author)

Simultaneous fluorescence light-up and selective multicolor nucleobase recognition based on sequence-dependent strong binding of berberine to DNA abasic site.

Science.gov (United States)

Wu, Fei; Shao, Yong; Ma, Kun; Cui, Qinghua; Liu, Guiying; Xu, Shujuan

2012-04-28

Label-free DNA nucleobase recognition by fluorescent small molecules has received much attention due to its simplicity in mutation identification and drug screening. However, sequence-dependent fluorescence light-up nucleobase recognition and multicolor emission with individual emission energy for individual nucleobases have been seldom realized. Herein, an abasic site (AP site) in a DNA duplex was employed as a binding field for berberine, one of isoquinoline alkaloids. Unlike weak binding of berberine to the fully matched DNAs without the AP site, strong binding of berberine to the AP site occurs and the berberine's fluorescence light-up behaviors are highly dependent on the target nucleobases opposite the AP site in which the targets thymine and cytosine produce dual emission bands, while the targets guanine and adenine only give a single emission band. Furthermore, more intense emissions are observed for the target pyrimidines than purines. The flanking bases of the AP site also produce some modifications of the berberine's emission behavior. The binding selectivity of berberine at the AP site is also confirmed by measurements of fluorescence resonance energy transfer, excited-state lifetime, DNA melting and fluorescence quenching by ferrocyanide and sodium chloride. It is expected that the target pyrimidines cause berberine to be stacked well within DNA base pairs near the AP site, which results in a strong resonance coupling of the electronic transitions to the particular vibration mode to produce the dual emissions. The fluorescent signal-on and emission energy-modulated sensing for nucleobases based on this fluorophore is substantially advantageous over the previously used fluorophores. We expect that this approach will be developed as a practical device for differentiating pyrimidines from purines by positioning an AP site toward a target that is available for readout by this alkaloid probe. This journal is © The Royal Society of Chemistry 2012

Towards observing the encounter of the T7 DNA replication fork with a lesion site at the Single molecule level

KAUST Repository

Shirbini, Afnan

2017-05-01

Single-molecule DNA flow-stretching assays have been a powerful approach to study various aspects on the mechanism of DNA replication for more than a decade. This technique depends on flow-induced force on a bead attached to a surface-tethered DNA. The difference in the elastic property between double-strand DNA (long) and single-strand DNA (short) at low regime force allows the observation of the beads motion when the dsDNA is converted to ssDNA by the replisome machinery during DNA replication. Here, I aim to develop an assay to track in real-time the encounter of the bacteriophage T7 replisome with abasic lesion site inserted on the leading strand template. I optimized methods to construct the DNA substrate that contains the abasic site and established the T7 leading strand synthesis at the single molecule level. I also optimized various control experiments to remove any interference from the nonspecific interactions of the DNA with the surface. My work established the foundation to image the encounter of the T7 replisome with abasic site and to characterize how the interactions between the helicase and the polymerase could influence the polymerase proofreading ability and its direct bypass of this highly common DNA damage type.

Stalled repair of lesions when present within a clustered DNA damage site

International Nuclear Information System (INIS)

Lomax, M.E.; Cunniffe, S.; O'Neill, P.

2003-01-01

Ionising radiation produces clustered DNA damages (two or more lesions within one or two helical turns of the DNA) which could challenge the repair mechanism(s) of the cell. Using purified base excision repair (BER) enzymes and synthetic oligonucleotides a number of recent studies have established the excision of a lesion within clustered damage sites is compromised. Evidence will be presented that the efficiency of repair of lesions within a clustered DNA damage site is reduced, relative to that of the isolated lesions, since the lifetime of both lesions is extended by up to four fold. Simple clustered damage sites, comprised of single-strand breaks, abasic sites and base damages, one or five bases 3' or 5' to each other, were synthesised in oligonucleotides and repair carried out in mammalian cell nuclear extracts. The rate of repair of the single-strand break/abasic site within these clustered damage sites is reduced, mainly due to inhibition of the DNA ligase. The mechanism of repair of the single-strand break/abasic site shows some asymmetry. Repair appears to be by the short-patch BER pathway when the lesions are 5' to each other. In contrast, when the lesions are 3' to each other repair appears to proceed along the long-patch BER pathway. The lesions within the cluster are processed sequentially, the single-strand break/abasic site being repaired before excision of 8-oxoG, limiting the formation of double-strand breaks to <2%. Stalled processing of clustered DNA damage extends the lifetime of the lesions to an extent that could have biological consequences, e.g. if the lesions are still present during transcription and/or at replication mutations could arise

Detecting single-abasic residues within a DNA strand immobilized in a biological nanopore using an integrated CMOS sensor.

Science.gov (United States)

Kim, Jungsuk; Maitra, Raj D; Pedrotti, Ken; Dunbar, William B

2013-02-01

In this paper, we demonstrate the application of a novel current-measuring sensor (CMS) customized for nanopore applications. The low-noise CMS is fabricated in a 0.35μm CMOS process and is implemented in experiments involving DNA captured in an α-hemolysin (α-HL) nanopore. Specifically, the CMS is used to build a current amplitude map as a function of varying positions of a single-abasic residue within a homopolymer cytosine single-stranded DNA (ssDNA) that is captured and held in the pore. Each ssDNA is immobilized using a biotin-streptavidin linkage. Five different DNA templates are measured and compared: one all-cytosine ssDNA, and four with a single-abasic residue substitution that resides in or near the ~1.5nm aperture of the α-HL channel when the strand is immobilized. The CMOS CMS is shown to resolves the ~5Å displacements of the abasic residue within the varying templates. The demonstration represents an advance in application-specific circuitry that is optimized for small-footprint nanopore applications, including genomic sequencing.

Analysis of translesion DNA synthesis activity of the human REV1 protein, which is a key player in radiation-induced mutagenesis

International Nuclear Information System (INIS)

Masuda, Yuji; Kamiya, Kenji

2003-01-01

Ionizing radiation frequently causes oxidative DNA damage in cells. It has been suggested that functions of the REV1 gene are induction of mutations and prevention of cell death caused by ionizing radiation through the damage bypass DNA replication. The gene product possesses a deoxycytidyl transferase activity, which is required for translesion DNA synthesis of a variety of damaged bases and an abasic site. To elucidate molecular mechanisms of the mutagenesis and translesion DNA synthesis, it is important to characterize the enzymatic properties of the REV1 protein. Here, we describe a novel method for purifying the recombinant human REV1 protein and the anzymatic properties of the protein. We established an efficient system for induction of the recombinant human REV1 protein in Escherichia coli cells. The REV1 protein was purified to homogeneity using nickel-chelating sepharose, heparin sepharose and superdex 200 chromatography. When purified by this method, REV1 protein is free of endo-, exonuclease and DNA polymerase activities. The purified REV1 protein is suitable for enzymological studies, and we used this to biochemical characterization. The REV1 protein inserts dCMP opposite templates G, A, T, C and an abasic site and inserts dGMP and dTMP opposite template G. Kinetic analysis provided evidence for high efficiency for dCMP insertion opposite template G and an abasic site, suggesting that the REV1 protein play a role in translesion DNA synthesis of an abasic site. (author)

Association between mutation spectra and stable and unstable DNA adduct profiles in Salmonella for benzo[a]pyrene and dibenzo[a,l]pyrene

Energy Technology Data Exchange (ETDEWEB)

DeMarini, David M., E-mail: demarini.david@epa.gov [Integrated Systems Toxicology Division, U.S. Environmental Protection Agency, Research Triangle Park, NC 27711 (United States); Hanley, Nancy M.; Warren, Sarah H.; Adams, Linda D.; King, Leon C. [Integrated Systems Toxicology Division, U.S. Environmental Protection Agency, Research Triangle Park, NC 27711 (United States)

2011-09-01

Highlights: {yields} Benzo[a]pyrene (BP) induces stable DNA adducts and mutations primarily at guanine. {yields} Dibenzo[a,l]pyrene (DBP) induces them primarily at adenine. {yields} BP induces abasic sites, but DBP does not in the Salmonella mutagenicity assay. {yields} Stable DNA adducts alone appear to account for the mutation spectrum of DBP. {yields} Stable DNA adducts and possibly abasic sites account for the mutation spectrum of BP. - Abstract: Benzo[a]pyrene (BP) and dibenzo[a,l]pyrene (DBP) are two polycyclic aromatic hydrocarbons (PAHs) that exhibit distinctly different mutagenicity and carcinogenicity profiles. Although some studies show that these PAHs produce unstable DNA adducts, conflicting data and arguments have been presented regarding the relative roles of these unstable adducts versus stable adducts, as well as oxidative damage, in the mutagenesis and tumor-mutation spectra of these PAHs. However, no study has determined the mutation spectra along with the stable and unstable DNA adducts in the same system with both PAHs. Thus, we determined the mutagenic potencies and mutation spectra of BP and DBP in strains TA98, TA100 and TA104 of Salmonella, and we also measured the levels of abasic sites (aldehydic-site assay) and characterized the stable DNA adducts ({sup 32}P-postlabeling/HPLC) induced by these PAHs in TA104. Our results for the mutation spectra and site specificity of stable adducts were consistent with those from other systems, showing that DBP was more mutagenic than BP in TA98 and TA100. The mutation spectra of DBP and BP were significantly different in TA98 and TA104, with 24% of the mutations induced by BP in TA98 being complex frameshifts, whereas DBP produced hardly any of these mutations. In TA104, BP produced primarily GC to TA transversions, whereas DBP produced primarily AT to TA transversions. The majority (96%) of stable adducts induced by BP were at guanine, whereas the majority (80%) induced by DBP were at adenine

On the Formation and Properties of Interstrand DNA-DNA Cross-links Forged by Reaction of an Abasic Site With the Opposing Guanine Residue of 5′-CAp Sequences in Duplex DNA

Science.gov (United States)

Johnson, Kevin M.; Price, Nathan E.; Wang, Jin; Fekry, Mostafa I.; Dutta, Sanjay; Seiner, Derrick R.; Wang, Yinsheng; Gates, Kent S.

2014-01-01

We recently reported that the aldehyde residue of an abasic (Ap) site in duplex DNA can generate an interstrand cross-link via reaction with a guanine residue on the opposing strand. This finding is intriguing because the highly deleterious nature of interstrand cross-links suggests that even small amounts of Ap-derived cross-links could make a significant contribution to the biological consequences stemming from the generation of Ap sites in cellular DNA. Incubation of 21-bp duplexes containing a central 5′-CAp sequence under conditions of reductive amination (NaCNBH3, pH 5.2) generated much higher yields of cross-linked DNA than reported previously. At pH 7, in the absence of reducing agents, these Ap-containing duplexes also produced cross-linked duplexes that were readily detected on denaturing polyacrylamide gels. Cross-link formation was not highly sensitive to reaction conditions and, once formed, the cross-link was stable to a variety of work-up conditions. Results of multiple experiments including MALDI-TOF mass spectrometry, gel mobility, methoxyamine capping of the Ap aldehyde, inosine-for-guanine replacement, hydroxyl radical footprinting, and LCMS/MS were consistent with a cross-linking mechanism involving reversible reaction of the Ap aldehyde residue with the N2-amino group of the opposing guanine residue in 5′-CAp sequences to generate hemiaminal, imine, or cyclic hemiaminal cross-links (7-10) that were irreversibly converted under conditions of reductive amination (NaCNBH3/pH 5.2) to a stable amine linkage. Further support for the importance of the exocyclic N2-amino group in this reaction was provided by an experiment showing that installation of a 2-aminopurine-thymine base pair at the cross-linking site produced high yields (15-30%) of a cross-linked duplex at neutral pH, in the absence of NaCNBH3. PMID:23215239

Chemical repair activity of free radical scavenger edaravone. Reduction reactions with dGMP hydroxyl radical adducts and suppression of base lesions and AP sites on irradiated plasmid DNA

International Nuclear Information System (INIS)

Hata, Kuniki; Katsumura, Yosuke; Urushibara, Ayumi; Yamashita, Shinichi; Lin Mingzhang; Muroya, Yusa; Shikazono, Naoya; Yokoya, Akinari; Fu Haiying

2015-01-01

Reactions of edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one) with deoxyguanosine monophosphate (dGMP) hydroxyl radical adducts were investigated by pulse radiolysis technique. Edaravone was found to reduce the dGMP hydroxyl radical adducts through electron transfer reactions. The rate constants of the reactions were greater than 4 × 10 8 dm 3 mol -1 s -1 and similar to those of the reactions of ascorbic acid, which is a representative antioxidant. Yields of single-strand breaks, base lesions, and abasic sites produced in pUC18 plasmid DNA by gamma ray irradiation in the presence of low concentrations (10–1000 μmol dm -3 ) of edaravone were also quantified, and the chemical repair activity of edaravone was estimated by a method recently developed by the authors. By comparing suppression efficiencies to the induction of each DNA lesion, it was found that base lesions and abasic sites were suppressed by the chemical repair activity of edaravone, although the suppression of single-strand breaks was not very effective. This phenomenon was attributed to the chemical repair activity of edaravone toward base lesions and abasic sites. However, the chemical repair activity of edaravone for base lesions was lower than that of ascorbic acid. (author)

Chemical repair activity of free radical scavenger edaravone: reduction reactions with dGMP hydroxyl radical adducts and suppression of base lesions and AP sites on irradiated plasmid DNA.

Science.gov (United States)

Hata, Kuniki; Urushibara, Ayumi; Yamashita, Shinichi; Lin, Mingzhang; Muroya, Yusa; Shikazono, Naoya; Yokoya, Akinari; Fu, Haiying; Katsumura, Yosuke

2015-01-01

Reactions of edaravone (3-methyl-1-phenyl-2-pyrazolin-5-one) with deoxyguanosine monophosphate (dGMP) hydroxyl radical adducts were investigated by pulse radiolysis technique. Edaravone was found to reduce the dGMP hydroxyl radical adducts through electron transfer reactions. The rate constants of the reactions were greater than 4 × 10(8) dm(3) mol(-1) s(-1) and similar to those of the reactions of ascorbic acid, which is a representative antioxidant. Yields of single-strand breaks, base lesions, and abasic sites produced in pUC18 plasmid DNA by gamma ray irradiation in the presence of low concentrations (10-1000 μmol dm(-3)) of edaravone were also quantified, and the chemical repair activity of edaravone was estimated by a method recently developed by the authors. By comparing suppression efficiencies to the induction of each DNA lesion, it was found that base lesions and abasic sites were suppressed by the chemical repair activity of edaravone, although the suppression of single-strand breaks was not very effective. This phenomenon was attributed to the chemical repair activity of edaravone toward base lesions and abasic sites. However, the chemical repair activity of edaravone for base lesions was lower than that of ascorbic acid. © The Author 2014. Published by Oxford University Press on behalf of The Japan Radiation Research Society and Japanese Society for Radiation Oncology.

Clustered DNA damages induced in human hematopoietic cells by low doses of ionizing radiation

Science.gov (United States)

Sutherland, Betsy M.; Bennett, Paula V.; Cintron-Torres, Nela; Hada, Megumi; Trunk, John; Monteleone, Denise; Sutherland, John C.; Laval, Jacques; Stanislaus, Marisha; Gewirtz, Alan

2002-01-01

Ionizing radiation induces clusters of DNA damages--oxidized bases, abasic sites and strand breaks--on opposing strands within a few helical turns. Such damages have been postulated to be difficult to repair, as are double strand breaks (one type of cluster). We have shown that low doses of low and high linear energy transfer (LET) radiation induce such damage clusters in human cells. In human cells, DSB are about 30% of the total of complex damages, and the levels of DSBs and oxidized pyrimidine clusters are similar. The dose responses for cluster induction in cells can be described by a linear relationship, implying that even low doses of ionizing radiation can produce clustered damages. Studies are in progress to determine whether clusters can be produced by mechanisms other than ionizing radiation, as well as the levels of various cluster types formed by low and high LET radiation.

Delayed repair of radiation induced clustered DNA damage: Friend or foe?

International Nuclear Information System (INIS)

Eccles, Laura J.; O'Neill, Peter; Lomax, Martine E.

2011-01-01

A signature of ionizing radiation exposure is the induction of DNA clustered damaged sites, defined as two or more lesions within one to two helical turns of DNA by passage of a single radiation track. Clustered damage is made up of double strand breaks (DSB) with associated base lesions or abasic (AP) sites, and non-DSB clusters comprised of base lesions, AP sites and single strand breaks. This review will concentrate on the experimental findings of the processing of non-DSB clustered damaged sites. It has been shown that non-DSB clustered damaged sites compromise the base excision repair pathway leading to the lifetime extension of the lesions within the cluster, compared to isolated lesions, thus the likelihood that the lesions persist to replication and induce mutation is increased. In addition certain non-DSB clustered damaged sites are processed within the cell to form additional DSB. The use of E. coli to demonstrate that clustering of DNA lesions is the major cause of the detrimental consequences of ionizing radiation is also discussed. The delayed repair of non-DSB clustered damaged sites in humans can be seen as a 'friend', leading to cell killing in tumour cells or as a 'foe', resulting in the formation of mutations and genetic instability in normal tissue.

DNA sequence analysis of spontaneous and γ-radiation (anoxic)-induced lacId mutations in Escherichia coli umuC122::Tn5: Differential requirement for umuC at G·C vs. A·T sites and for the production of transversions vs. transitions

International Nuclear Information System (INIS)

Sargentini, Neil J.; Smith, Kendric C.

1994-01-01

Escherichia coli umuC122::Tn5 cells were γ-irradiated ( 137 Cs, 750 Gy, under N 2 ), and lac-constitutive mutants were produced at 36% of the wild-type level. The specific nature of the umuC strain's partial radiation mutability was determined by sequencing 325 radiation-induced lacI d mutations. The yields of radiation-induced mutation classes in the umuC strain (as a percentage of the wild-type yield) were: 80% for A·T approaches G·C transitions, 70% for multi-base additions, 60% for single-base deletions, 53% for A·T approaches C·G transversions, 36% for G·C approaches A·T transitions, 25% for multi-base deletions, 21% for A·T approaches T·A transversions, 11% for G·C approaches C·G transversions, 9% for G·C approaches T·A transversions, and 0% for multiple mutations. Based on these deficiencies and other factors, it is concluded that the umuC strain is near-normal for A·T approaches G·C transitions, single-base deletions and possibly A·T approaches C·G transversions; is generally deficient for mutagenesis at G·C sites and for transversions, and is grossly deficient in multiple mutations. Damage at G·C sites seems more difficult for translesion DNA synthesis to bypass than damage at A·T sites, and especially when trying to produce a transversion. The yield of G·C approaches A·T transitions in the umuC strain (36% of the wild-type level) argues that abasic sites are involved in no more than 64% of γ-radiation-induced base substitutions in the wild-type strain. Altogether, these data suggest that the UmuC and UmuD' proteins facilitate, rather than being absolutely required for, translesion DNA synthesis; with the degree of facilitation being dependent both on the nature of the non-coding DNA damage, i.e., at G·C vs. A·T sites, and on the nature of the mis-incorporated base, i.e., whether it induces transversions or transitions

Delayed repair of radiation induced clustered DNA damage: Friend or foe?

Science.gov (United States)

Eccles, Laura J.; O’Neill, Peter; Lomax, Martine E.

2011-01-01

A signature of ionizing radiation exposure is the induction of DNA clustered damaged sites, defined as two or more lesions within one to two helical turns of DNA by passage of a single radiation track. Clustered damage is made up of double strand breaks (DSB) with associated base lesions or abasic (AP) sites, and non-DSB clusters comprised of base lesions, AP sites and single strand breaks. This review will concentrate on the experimental findings of the processing of non-DSB clustered damaged sites. It has been shown that non-DSB clustered damaged sites compromise the base excision repair pathway leading to the lifetime extension of the lesions within the cluster, compared to isolated lesions, thus the likelihood that the lesions persist to replication and induce mutation is increased. In addition certain non-DSB clustered damaged sites are processed within the cell to form additional DSB. The use of E. coli to demonstrate that clustering of DNA lesions is the major cause of the detrimental consequences of ionizing radiation is also discussed. The delayed repair of non-DSB clustered damaged sites in humans can be seen as a “friend”, leading to cell killing in tumour cells or as a “foe”, resulting in the formation of mutations and genetic instability in normal tissue. PMID:21130102

Effect of the bases flanking an abasic site on the recognition of nucleobase by amiloride.

Science.gov (United States)

Rajendran, Arivazhagan; Zhao, Chunxia; Rajendar, Burki; Thiagarajan, Viruthachalam; Sato, Yusuke; Nishizawa, Seiichi; Teramae, Norio

2010-06-01

We explain here the various non-covalent interactions which are responsible for the different binding modes of a small ligand with DNA. The combination of experimental and theoretical methods was used. The interaction of amiloride with thymine was found to depend on the bases flanking the AP site and different binding modes were observed for different flanking bases. Molecular modeling, absorption studies and binding constant measurements support for the different binding patterns. The flanking base dependent recognition of AP site phosphates was investigated by (31)P NMR experiments. The thermodynamics of the ligand-nucleotide interaction was demonstrated by isothermal titration calorimetry. The emission behavior of amiloride was found to depend on the bases flanking the AP site. Amiloride photophysics in the context of AP-site containing DNA is investigated by time-dependent density functional theory. Flanking bases affect the ground and excited electronic states of amiloride when binding to AP site, which causes flanking base-dependent fluorescence signaling. The various noncovalent interactions have been well characterized for the determination of nucleic acid structure and dynamics, and protein-DNA interactions. However, these are not clear for the DNA-small molecule interactions and we believe that our studies will bring a new insight into such phenomena. Copyright 2010 Elsevier B.V. All rights reserved.

In vitro selection of shape-changing DNA nanostructures capable of binding-induced cargo release.

Science.gov (United States)

Oh, Seung Soo; Plakos, Kory; Xiao, Yi; Eisenstein, Michael; Soh, H Tom

2013-11-26

Many biological systems employ allosteric regulatory mechanisms, which offer a powerful means of directly linking a specific binding event to a wide spectrum of molecular functionalities. There is considerable interest in generating synthetic allosteric regulators that can perform useful molecular functions for applications in diagnostics, imaging and targeted therapies, but generating such molecules through either rational design or directed evolution has proven exceptionally challenging. To address this need, we present an in vitro selection strategy for generating conformation-switching DNA nanostructures that selectively release a small-molecule payload in response to binding of a specific trigger molecule. As an exemplar, we have generated a DNA nanostructure that hybridizes with a separate 'cargo strand' containing an abasic site. This abasic site stably sequesters a fluorescent cargo molecule in an inactive state until the DNA nanostructure encounters an ATP trigger molecule. This ATP trigger causes the nanostructure to release the cargo strand, thereby liberating the fluorescent payload and generating a detectable fluorescent readout. Our DNA nanostructure is highly sensitive, with an EC50 of 30 μM, and highly specific, releasing its payload in response to ATP but not to other chemically similar nucleotide triphosphates. We believe that this selection approach could be generalized to generate synthetic nanostructures capable of selective and controlled release of other small-molecule cargos in response to a variety of triggers, for both research and clinical applications.

DNA-dependent protein kinase inhibits AID-induced antibody gene conversion.

Directory of Open Access Journals (Sweden)

Adam J L Cook

2007-04-01

Full Text Available Affinity maturation and class switching of antibodies requires activation-induced cytidine deaminase (AID-dependent hypermutation of Ig V(DJ rearrangements and Ig S regions, respectively, in activated B cells. AID deaminates deoxycytidine bases in Ig genes, converting them into deoxyuridines. In V(DJ regions, subsequent excision of the deaminated bases by uracil-DNA glycosylase, or by mismatch repair, leads to further point mutation or gene conversion, depending on the species. In Ig S regions, nicking at the abasic sites produced by AID and uracil-DNA glycosylases results in staggered double-strand breaks, whose repair by nonhomologous end joining mediates Ig class switching. We have tested whether nonhomologous end joining also plays a role in V(DJ hypermutation using chicken DT40 cells deficient for Ku70 or the DNA-dependent protein kinase catalytic subunit (DNA-PKcs. Inactivation of the Ku70 or DNA-PKcs genes in DT40 cells elevated the rate of AID-induced gene conversion as much as 5-fold. Furthermore, DNA-PKcs-deficiency appeared to reduce point mutation. The data provide strong evidence that double-strand DNA ends capable of recruiting the DNA-dependent protein kinase complex are important intermediates in Ig V gene conversion.

Theoretical approach of complex DNA lesions: from formation to repair

International Nuclear Information System (INIS)

Bignon, Emmanuelle

2017-01-01

This thesis work is focused on the theoretical modelling of DNA damages, from formation to repair. Several projects have been led in this framework, which can be sorted into three different parts. One on hand, we studied complex DNA reactivity. It included a study about 8-oxo-7,8-dihydro-guanine (8oxoG) mechanisms of formation, a project concerning the UV-induced pyrimidine 6-4 pyrimidone (6-4PP) endogenous photo-sensitizer features, and another one about DNA photo-sensitization by nonsteroidal anti-inflammatory drugs (i.e. ketoprofen and ibuprofen). On the other hand, we investigated mechanical properties of damaged DNA. The structural signature of a DNA lesion is of major importance for their repair, unfortunately only few NMR and X-ray structures of such systems are available. In order to gain insights into their dynamical structure, we investigated a series of complex damages: clustered abasic sites, interstrand cross-links, and the 6-4PP photo-lesion. Likewise, we studied the interaction modes DNA with several polyamines, which are well known to interact with the double helix, but also with the perspective to model DNA-protein cross-linking. The third part concerned the study of DNA interactions with repair enzymes. In line with the structural study about clustered abasic sites, we investigated the dynamics of the same system, but this time interacting with the APE1 endonuclease. We also studied interactions between the Fpg glycosylase with an oligonucleotides containing tandem 8-oxoG on one hand and 8-oxoG - abasic site as multiply damaged sites. Thus, we shed new lights on damaged DNA reactivity, structure and repair, which provides perspectives for biomedicine and life's mechanisms understanding as we begin to describe nucleosomal DNA. (author)

Non-Covalent Fluorescent Labeling of Hairpin DNA Probe Coupled with Hybridization Chain Reaction for Sensitive DNA Detection.

Science.gov (United States)

Song, Luna; Zhang, Yonghua; Li, Junling; Gao, Qiang; Qi, Honglan; Zhang, Chengxiao

2016-04-01

An enzyme-free signal amplification-based assay for DNA detection was developed using fluorescent hairpin DNA probes coupled with hybridization chain reaction (HCR). The hairpin DNAs were designed to contain abasic sites in the stem moiety. Non-covalent labeling of the hairpin DNAs was achieved when a fluorescent ligand was bound to the abasic sites through hydrogen bonding with the orphan cytosine present on the complementary strand, accompanied by quench of ligand fluorescence. As a result, the resultant probes, the complex formed between the hairpin DNA and ligand, showed almost no fluorescence. Upon hybridization with target DNA, the probe underwent a dehybridization of the stem moiety containing an abasic site. The release of ligand from the abasic site to the solution resulted in an effective fluorescent enhancement, which can be used as a signal. Compared with a sensing system without HCR, a 20-fold increase in the sensitivity was achieved using the sensing system with HCR. The fluorescent intensity of the sensing system increased with the increase in target DNA concentration from 0.5 nM to 100 nM. A single mismatched target ss-DNA could be effectively discriminated from complementary target DNA. Genotyping of a G/C single-nucleotide polymorphism of polymerase chain reaction (PCR) products was successfully demonstrated with the sensing system. Therefore, integrating HCR strategy with non-covalent labeling of fluorescent hairpin DNA probes provides a sensitive and cost-effective DNA assay. © The Author(s) 2016.

Yields of clustered DNA damage induced by charged-particle radiations of similar kinetic energy per nucleon: LET dependence in different DNA microenvironments

International Nuclear Information System (INIS)

Keszenman, D.J.; Sutherland, B.M.

2010-01-01

To determine the linear energy transfer (LET) dependence of the biological effects of densely ionizing radiation in relation to changes in the ionization density along the track, we measured the yields and spectrum of clustered DNA damages induced by charged particles of different atomic number but similar kinetic energy per nucleon in different DNA microenvironments. Yeast DNA embedded in agarose in solutions of different free radical scavenging capacity was irradiated with 1 GeV protons, 1 GeV/nucleon oxygen ions, 980 MeV/nucleon titanium ions or 968 MeV/nucleon iron ions. The frequencies of double-strand breaks (DSBs), abasic sites and oxypurine clusters were quantified. The total DNA damage yields per absorbed dose induced in non-radioquenching solution decreased with LET, with minor variations in radioquenching conditions being detected. However, the total damage yields per particle fluence increased with LET in both conditions, indicating a higher efficiency per particle to induce clustered DNA damages. The yields of DSBs and non-DSB clusters as well as the damage spectra varied with LET and DNA milieu, suggesting the involvement of more than one mechanism in the formation of the different types of clustered damages.

Aag-initiated base excision repair promotes ischemia reperfusion injury in liver, brain, and kidney.

Science.gov (United States)

Ebrahimkhani, Mohammad R; Daneshmand, Ali; Mazumder, Aprotim; Allocca, Mariacarmela; Calvo, Jennifer A; Abolhassani, Nona; Jhun, Iny; Muthupalani, Sureshkumar; Ayata, Cenk; Samson, Leona D

2014-11-11

Inflammation is accompanied by the release of highly reactive oxygen and nitrogen species (RONS) that damage DNA, among other cellular molecules. Base excision repair (BER) is initiated by DNA glycosylases and is crucial in repairing RONS-induced DNA damage; the alkyladenine DNA glycosylase (Aag/Mpg) excises several DNA base lesions induced by the inflammation-associated RONS release that accompanies ischemia reperfusion (I/R). Using mouse I/R models we demonstrate that Aag(-/-) mice are significantly protected against, rather than sensitized to, I/R injury, and that such protection is observed across three different organs. Following I/R in liver, kidney, and brain, Aag(-/-) mice display decreased hepatocyte death, cerebral infarction, and renal injury relative to wild-type. We infer that in wild-type mice, Aag excises damaged DNA bases to generate potentially toxic abasic sites that in turn generate highly toxic DNA strand breaks that trigger poly(ADP-ribose) polymerase (Parp) hyperactivation, cellular bioenergetics failure, and necrosis; indeed, steady-state levels of abasic sites and nuclear PAR polymers were significantly more elevated in wild-type vs. Aag(-/-) liver after I/R. This increase in PAR polymers was accompanied by depletion of intracellular NAD and ATP levels plus the translocation and extracellular release of the high-mobility group box 1 (Hmgb1) nuclear protein, activating the sterile inflammatory response. We thus demonstrate the detrimental effects of Aag-initiated BER during I/R and sterile inflammation, and present a novel target for controlling I/R-induced injury.

Functional Sites Induce Long-Range Evolutionary Constraints in Enzymes.

Directory of Open Access Journals (Sweden)

Benjamin R Jack

2016-05-01

Full Text Available Functional residues in proteins tend to be highly conserved over evolutionary time. However, to what extent functional sites impose evolutionary constraints on nearby or even more distant residues is not known. Here, we report pervasive conservation gradients toward catalytic residues in a dataset of 524 distinct enzymes: evolutionary conservation decreases approximately linearly with increasing distance to the nearest catalytic residue in the protein structure. This trend encompasses, on average, 80% of the residues in any enzyme, and it is independent of known structural constraints on protein evolution such as residue packing or solvent accessibility. Further, the trend exists in both monomeric and multimeric enzymes and irrespective of enzyme size and/or location of the active site in the enzyme structure. By contrast, sites in protein-protein interfaces, unlike catalytic residues, are only weakly conserved and induce only minor rate gradients. In aggregate, these observations show that functional sites, and in particular catalytic residues, induce long-range evolutionary constraints in enzymes.

UVA-induced DNA double-strand breaks result from the repair of clustered oxidative DNA damages

Science.gov (United States)

Greinert, R.; Volkmer, B.; Henning, S.; Breitbart, E. W.; Greulich, K. O.; Cardoso, M. C.; Rapp, Alexander

2012-01-01

UVA (320–400 nm) represents the main spectral component of solar UV radiation, induces pre-mutagenic DNA lesions and is classified as Class I carcinogen. Recently, discussion arose whether UVA induces DNA double-strand breaks (dsbs). Only few reports link the induction of dsbs to UVA exposure and the underlying mechanisms are poorly understood. Using the Comet-assay and γH2AX as markers for dsb formation, we demonstrate the dose-dependent dsb induction by UVA in G1-synchronized human keratinocytes (HaCaT) and primary human skin fibroblasts. The number of γH2AX foci increases when a UVA dose is applied in fractions (split dose), with a 2-h recovery period between fractions. The presence of the anti-oxidant Naringin reduces dsb formation significantly. Using an FPG-modified Comet-assay as well as warm and cold repair incubation, we show that dsbs arise partially during repair of bi-stranded, oxidative, clustered DNA lesions. We also demonstrate that on stretched chromatin fibres, 8-oxo-G and abasic sites occur in clusters. This suggests a replication-independent formation of UVA-induced dsbs through clustered single-strand breaks via locally generated reactive oxygen species. Since UVA is the main component of solar UV exposure and is used for artificial UV exposure, our results shine new light on the aetiology of skin cancer. PMID:22941639

Cytosine hypomethylation at CHG and CHH sites in the pleiotropic ...

Indian Academy of Sciences (India)

2013-12-16

Dec 16, 2013 ... The abasic gap left behind by the demethylase action is filled by any of the DNA ...... Inactivation of a DNA methylation pathway in maize reproduc- tive organs ... of diversity for morphological and yield related traits among.

Relationship between hydrogen-induced phase transformations and pitting nucleation sites in duplex stainless steel

Energy Technology Data Exchange (ETDEWEB)

Guo, Liqiu; Yang, Binjie; Qin, Sixiao [University of Science and Technology Beijing (China). Corrosion and Protection Center

2016-02-15

This paper demonstrates the hydrogen-induced phase transformation and the associated pitting nucleation sites of 2507 duplex stainless steel using scanning Kelvin probe force microscopy and magnetic force microscopy. The low potential sites in Volta potential images, which are considered as the pitting nucleation sites, are strongly dependent on the hydrogen-induced phase transformation. They firstly initiate on the magnetic martensite laths in the austenite phase or at the ferrite/austenite boundaries, and then appear near the needle-shaped microtwins in the ferrite phase, because of the difference in physicochemical properties of hydrogen-induced phase transformation microstructures.

Site-controlled fabrication of silicon nanotips by indentation-induced selective etching

Science.gov (United States)

Jin, Chenning; Yu, Bingjun; Liu, Xiaoxiao; Xiao, Chen; Wang, Hongbo; Jiang, Shulan; Wu, Jiang; Liu, Huiyun; Qian, Linmao

2017-12-01

In the present study, the indentation-induced selective etching approach is proposed to fabricate site-controlled pyramidal nanotips on Si(100) surface. Without any masks, the site-controlled nanofabrication can be realized by nanoindentation and post etching in potassium hydroxide (KOH) solution. The effect of indentation force and etching time on the formation of pyramidal nanotips was investigated. It is found that the height and radius of the pyramidal nanotips increase with the indentation force or etching time, while long-time etching can lead to the collapse of the tips. The formation of pyramidal tips is ascribed to the anisotropic etching of silicon and etching stop of (111) crystal planes in KOH aqueous solution. The capability of this fabrication method was further demonstrated by producing various tip arrays on silicon surface by selective etching of the site-controlled indent patterns, and the maximum height difference of these tips is less than 10 nm. The indentation-induced selective etching provides a new strategy to fabricate well site-controlled tip arrays for multi-probe SPM system, Si nanostructure-based sensors and high-quality information storage.

Analysis of Structural Flexibility of Damaged DNA Using Thiol-Tethered Oligonucleotide Duplexes.

Directory of Open Access Journals (Sweden)

Masashi Fujita

Full Text Available Bent structures are formed in DNA by the binding of small molecules or proteins. We developed a chemical method to detect bent DNA structures. Oligonucleotide duplexes in which two mercaptoalkyl groups were attached to the positions facing each other across the major groove were prepared. When the duplex contained the cisplatin adduct, which was proved to induce static helix bending, interstrand disulfide bond formation under an oxygen atmosphere was detected by HPLC analyses, but not in the non-adducted duplex, when the two thiol-tethered nucleosides were separated by six base pairs. When the insert was five and seven base pairs, the disulfide bond was formed and was not formed, respectively, regardless of the cisplatin adduct formation. The same reaction was observed in the duplexes containing an abasic site analog and the (6–4 photoproduct. Compared with the cisplatin case, the disulfide bond formation was slower in these duplexes, but the reaction rate was nearly independent of the linker length. These results indicate that dynamic structural changes of the abasic site- and (6–4 photoproduct-containing duplexes could be detected by our method. It is strongly suggested that the UV-damaged DNA-binding protein, which specifically binds these duplexes and functions at the first step of global-genome nucleotide excision repair, recognizes the easily bendable nature of damaged DNA.

Endonuclease IV Is the major apurinic/apyrimidinic endonuclease in Mycobacterium tuberculosis and is important for protection against oxidative damage.

Directory of Open Access Journals (Sweden)

Rupangi Verma Puri

Full Text Available During the establishment of an infection, bacterial pathogens encounter oxidative stress resulting in the production of DNA lesions. Majority of these lesions are repaired by base excision repair (BER pathway. Amongst these, abasic sites are the most frequent lesions in DNA. Class II apurinic/apyrimidinic (AP endonucleases play a major role in BER of damaged DNA comprising of abasic sites. Mycobacterium tuberculosis, a deadly pathogen, resides in the human macrophages and is continually subjected to oxidative assaults. We have characterized for the first time two AP endonucleases namely Endonuclease IV (End and Exonuclease III (XthA that perform distinct functions in M.tuberculosis. We demonstrate that M.tuberculosis End is a typical AP endonuclease while XthA is predominantly a 3'→5' exonuclease. The AP endonuclease activity of End and XthA was stimulated by Mg(2+ and Ca(2+ and displayed a preferential recognition for abasic site paired opposite to a cytosine residue in DNA. Moreover, End exhibited metal ion independent 3'→5' exonuclease activity while in the case of XthA this activity was metal ion dependent. We demonstrate that End is not only a more efficient AP endonuclease than XthA but it also represents the major AP endonuclease activity in M.tuberculosis and plays a crucial role in defense against oxidative stress.

A distant cis acting intronic element induces site-selective RNA editing

DEFF Research Database (Denmark)

Daniel, Chammiran; Venø, Morten Trillingsgaard; Ekdahl, Ylva

2012-01-01

Transcripts have been found to be site selectively edited from adenosine-to-inosine (A-to-I) in the mammalian brain, mostly in genes involved in neurotransmission. While A-to-I editing occurs at double-stranded structures, other structural requirements are largely unknown. We have investigated...... shown to be important for A-to-I editing. We demonstrate that the element also can induce editing in related but normally not edited RNA sequences. In human, thousands of genes are edited in duplexes formed by inverted repeats in non-coding regions. It is likely that numerous such duplexes can induce...... the requirements for editing at the I/M site in the Gabra-3 transcript of the GABA(A) receptor. We identify an evolutionarily conserved intronic duplex, 150 nt downstream of the exonic hairpin where the I/M site resides, which is required for its editing. This is the first time a distant RNA structure has been...

Melatonin Protects Human Cells from Clustered DNA Damages, Killing and Acquisition of Soft Agar Growth Induced by X-rays or 970 MeV/n Fe ions

Energy Technology Data Exchange (ETDEWEB)

Das, B.; Sutherland, B.; Bennett, P. V.; Cutter, N. C.; Sutherland, J. C.

2011-06-01

We tested the ability of melatonin (N-acetyl-5 methoxytryptamine), a highly effective radical scavenger and human hormone, to protect DNA in solution and in human cells against induction of complex DNA clusters and biological damage induced by low or high linear energy transfer radiation (100 kVp X-rays, 970 MeV/nucleon Fe ions). Plasmid DNA in solution was treated with increasing concentrations of melatonin (0.0-3.5 mM) and were irradiated with X-rays. Human cells (28SC monocytes) were also irradiated with X-rays and Fe ions with and without 2 mM melatonin. Agarose plugs containing genomic DNA were subjected to Contour Clamped Homogeneous Electrophoretic Field (CHEF) followed by imaging and clustered DNA damages were measured by using Number Average length analysis. Transformation experiments on human primary fibroblast cells using soft agar colony assay were carried out which were irradiated with Fe ions with or without 2 mM melatonin. In plasmid DNA in solution, melatonin reduced the induction of single- and double-strand breaks. Pretreatment of human 28SC cells for 24 h before irradiation with 2 mM melatonin reduced the level of X-ray induced double-strand breaks by {approx}50%, of abasic clustered damages about 40%, and of Fe ion-induced double-strand breaks (41% reduction) and abasic clusters (34% reduction). It decreased transformation to soft agar growth of human primary cells by a factor of 10, but reduced killing by Fe ions only by 20-40%. Melatonin's effective reduction of radiation-induced critical DNA damages, cell killing, and striking decrease of transformation suggest that it is an excellent candidate as a countermeasure against radiation exposure, including radiation exposure to astronaut crews in space travel.

Base excision repair of chemotherapeutically-induced alkylated DNA damage predominantly causes contractions of expanded GAA repeats associated with Friedreich's ataxia.

Directory of Open Access Journals (Sweden)

Yanhao Lai

Full Text Available Expansion of GAA·TTC repeats within the first intron of the frataxin gene is the cause of Friedreich's ataxia (FRDA, an autosomal recessive neurodegenerative disorder. However, no effective treatment for the disease has been developed as yet. In this study, we explored a possibility of shortening expanded GAA repeats associated with FRDA through chemotherapeutically-induced DNA base lesions and subsequent base excision repair (BER. We provide the first evidence that alkylated DNA damage induced by temozolomide, a chemotherapeutic DNA damaging agent can induce massive GAA repeat contractions/deletions, but only limited expansions in FRDA patient lymphoblasts. We showed that temozolomide-induced GAA repeat instability was mediated by BER. Further characterization of BER of an abasic site in the context of (GAA20 repeats indicates that the lesion mainly resulted in a large deletion of 8 repeats along with small expansions. This was because temozolomide-induced single-stranded breaks initially led to DNA slippage and the formation of a small GAA repeat loop in the upstream region of the damaged strand and a small TTC loop on the template strand. This allowed limited pol β DNA synthesis and the formation of a short 5'-GAA repeat flap that was cleaved by FEN1, thereby leading to small repeat expansions. At a later stage of BER, the small template loop expanded into a large template loop that resulted in the formation of a long 5'-GAA repeat flap. Pol β then performed limited DNA synthesis to bypass the loop, and FEN1 removed the long repeat flap ultimately causing a large repeat deletion. Our study indicates that chemotherapeutically-induced alkylated DNA damage can induce large contractions/deletions of expanded GAA repeats through BER in FRDA patient cells. This further suggests the potential of developing chemotherapeutic alkylating agents to shorten expanded GAA repeats for treatment of FRDA.

External human induced events in site evaluation for nuclear power plants. Safety guide

International Nuclear Information System (INIS)

2004-01-01

The purpose of the present Safety Guide is to provide recommendations and guidance for the examination of the region considered for site evaluation for a plant in order to identity hazardous phenomena associated with human induced events initiated by sources external to the plant. In some cases it also presents preliminary guidance for deriving values of relevant parameters for the design basis. This Safety Guide is also applicable for periodic site evaluation and site evaluation following a major human induced event, and for the design and operation of the site's environmental monitoring system. Site evaluation includes site characterization. Consideration of external events that could lead to a degradation of the safety features of the plant and cause a release of radioactive material from the plant and/or affect the dispersion of such material in the environment. And consideration of population issues and access issues significant to safety (such as the feasibility of evacuation, the population distribution and the location of resources). The process of site evaluation continues throughout the lifetime of the facility, from siting to design, construction, operation and decommissioning. The external human induced events considered in this Safety Guide are all of accidental origin. Considerations relating to the physical protection of the plant against wilful actions by third parties are outside its scope. However, the methods described herein may also have some application for the purposes of such physical protection. The present Safety Guide may also be used for events that may originate within the boundaries of the site, but from sources which are not directly involved in the operational states of the nuclear power plant units, such as fuel depots or areas for the storage of hazardous materials for the construction of other facilities at the same site. Special consideration should be given to the hazardous material handled during the construction, operation and

A novel method for monitoring functional lesion-specific recruitment of repair proteins in live cells

Energy Technology Data Exchange (ETDEWEB)

Woodrick, Jordan; Gupta, Suhani; Khatkar, Pooja; Dave, Kalpana; Levashova, Darya; Choudhury, Sujata; Elias, Hadi; Saha, Tapas; Mueller, Susette; Roy, Rabindra, E-mail: rr228@georgetown.edu

2015-05-15

Highlights: • A method of monitoring lesion-specific recruitment of proteins in vivo is described. • Recruitment of repair enzymes to abasic sites is monitored by co-localization. • Repair protein recruitment is consistent with known protein–protein relationships. • Cells demonstrated complete repair of abasic sites by 90 min. - Abstract: DNA–protein relationships have been studied by numerous methods, but a particular gap in methodology lies in the study of DNA adduct-specific interactions with proteins in vivo, which particularly affects the field of DNA repair. Using the repair of a well-characterized and ubiquitous adduct, the abasic (AP) site, as a model, we have developed a comprehensive method of monitoring DNA lesion-specific recruitment of proteins in vivo over time. We utilized a surrogate system in which a Cy3-labeled plasmid containing a single AP-site was transfected into cells, and the interaction of the labeled DNA with BER enzymes, including APE1, Polβ, LIG1, and FEN1, was monitored by immunofluorescent staining of the enzymes by Alexafluor-488-conjugated secondary antibody. The recruitment of enzymes was characterized by quantification of Cy3-Alexafluor-488 co-localization. To validate the microscopy-based method, repair of the transfected AP-site DNA was also quantified at various time points post-transfection using a real time PCR-based method. Notably, the recruitment time kinetics for each enzyme were consistent with AP-site repair time kinetics. This microscopy-based methodology is reliable in detecting the recruitment of proteins to specific DNA substrates and can be extended to study other in vivo DNA–protein relationships in any DNA sequence and in the context of any DNA structure in transfectable proliferating or quiescent cells. The method may be applied to a variety of disciplines of nucleic acid transaction pathways, including repair, replication, transcription, and recombination.

A novel method for monitoring functional lesion-specific recruitment of repair proteins in live cells

International Nuclear Information System (INIS)

Woodrick, Jordan; Gupta, Suhani; Khatkar, Pooja; Dave, Kalpana; Levashova, Darya; Choudhury, Sujata; Elias, Hadi; Saha, Tapas; Mueller, Susette; Roy, Rabindra

2015-01-01

Highlights: • A method of monitoring lesion-specific recruitment of proteins in vivo is described. • Recruitment of repair enzymes to abasic sites is monitored by co-localization. • Repair protein recruitment is consistent with known protein–protein relationships. • Cells demonstrated complete repair of abasic sites by 90 min. - Abstract: DNA–protein relationships have been studied by numerous methods, but a particular gap in methodology lies in the study of DNA adduct-specific interactions with proteins in vivo, which particularly affects the field of DNA repair. Using the repair of a well-characterized and ubiquitous adduct, the abasic (AP) site, as a model, we have developed a comprehensive method of monitoring DNA lesion-specific recruitment of proteins in vivo over time. We utilized a surrogate system in which a Cy3-labeled plasmid containing a single AP-site was transfected into cells, and the interaction of the labeled DNA with BER enzymes, including APE1, Polβ, LIG1, and FEN1, was monitored by immunofluorescent staining of the enzymes by Alexafluor-488-conjugated secondary antibody. The recruitment of enzymes was characterized by quantification of Cy3-Alexafluor-488 co-localization. To validate the microscopy-based method, repair of the transfected AP-site DNA was also quantified at various time points post-transfection using a real time PCR-based method. Notably, the recruitment time kinetics for each enzyme were consistent with AP-site repair time kinetics. This microscopy-based methodology is reliable in detecting the recruitment of proteins to specific DNA substrates and can be extended to study other in vivo DNA–protein relationships in any DNA sequence and in the context of any DNA structure in transfectable proliferating or quiescent cells. The method may be applied to a variety of disciplines of nucleic acid transaction pathways, including repair, replication, transcription, and recombination

N-terminal domains of human DNA polymerase lambda promote primer realignment during translesion DNA synthesis

Science.gov (United States)

Taggart, David J.; Dayeh, Daniel M.; Fredrickson, Saul W.; Suo, Zucai

2014-01-01

The X-family DNA polymerases λ (Polλ) and β (Polβ) possess similar 5′-2-deoxyribose-5-phosphatelyase (dRPase) and polymerase domains. Besides these domains, Polλ also possesses a BRCA1 C-terminal (BRCT) domain and a proline-rich domain at its N terminus. However, it is unclear how these non-enzymatic domains contribute to the unique biological functions of Polλ. Here, we used primer extension assays and a newly developed high-throughput short oligonucleotide sequencing assay (HT-SOSA) to compare the efficiency of lesion bypass and fidelity of human Polβ, Polλ and two N-terminal deletion constructs of Polλ during the bypass of either an abasic site or a 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) lesion. We demonstrate that the BRCT domain of Polλ enhances the efficiency of abasic site bypass by approximately 1.6-fold. In contrast, deletion of the N-terminal domains of Polλ did not affect the efficiency of 8-oxodG bypass relative to nucleotide incorporations opposite undamaged dG. HT-SOSA analysis demonstrated that Polλ and Polβ preferentially generated −1 or −2 frameshift mutations when bypassing an abasic site and the single or double base deletion frequency was highly sequence dependent. Interestingly, the BRCT and proline-rich domains of Polλ cooperatively promoted the generation of −2 frameshift mutations when the abasic site was situated within a sequence context that was susceptible to homology-driven primer realignment. Furthermore, both N-terminal domains of Polλ increased the generation of −1 frameshift mutations during 8-oxodG bypass and influenced the frequency of substitution mutations produced by Polλ opposite the 8-oxodG lesion. Overall, our data support a model wherein the BRCT and proline-rich domains of Polλ act cooperatively to promote primer/template realignment between DNA strands of limited sequence homology. This function of the N-terminal domains may facilitate the role of Polλ as a gap-filling polymerase

Stress-induced enhancement of leukocyte trafficking into sites of surgery or immune activation

Science.gov (United States)

Viswanathan, Kavitha; Dhabhar, Firdaus S.

2005-04-01

Effective immunoprotection requires rapid recruitment of leukocytes into sites of surgery, wounding, infection, or vaccination. In contrast to immunosuppressive chronic stressors, short-term acute stressors have immunoenhancing effects. Here, we quantify leukocyte infiltration within a surgical sponge to elucidate the kinetics, magnitude, subpopulation, and chemoattractant specificity of an acute stress-induced increase in leukocyte trafficking to a site of immune activation. Mice acutely stressed before sponge implantation showed 200-300% higher neutrophil, macrophage, natural killer cell, and T cell infiltration than did nonstressed animals. We also quantified the effects of acute stress on lymphotactin- (LTN; a predominantly lymphocyte-specific chemokine), and TNF-- (a proinflammatory cytokine) stimulated leukocyte infiltration. An additional stress-induced increase in infiltration was observed for neutrophils, in response to TNF-, macrophages, in response to TNF- and LTN, and natural killer cells and T cells in response to LTN. These results show that acute stress initially increases trafficking of all major leukocyte subpopulations to a site of immune activation. Tissue damage-, antigen-, or pathogen-driven chemoattractants subsequently determine which subpopulations are recruited more vigorously. Such stress-induced increases in leukocyte trafficking may enhance immunoprotection during surgery, vaccination, or infection, but may also exacerbate immunopathology during inflammatory (cardiovascular disease or gingivitis) or autoimmune (psoriasis, arthritis, or multiple sclerosis) diseases. chemokine | psychophysiological stress | surgical sponge | wound healing | lymphotactin

Seismic site characterization of an urban dedimentary basin, Livermore Valley, California: Site tesponse, basin-edge-induced surface waves, and 3D simulations

Science.gov (United States)

Hartzell, Stephen; Leeds, Alena L.; Ramirez-Guzman, Leonardo; Allen, James P.; Schmitt, Robert G.

2016-01-01

Thirty‐two accelerometers were deployed in the Livermore Valley, California, for approximately one year to study sedimentary basin effects. Many local and near‐regional earthquakes were recorded, including the 24 August 2014 Mw 6.0 Napa, California, earthquake. The resulting ground‐motion data set is used to quantify the seismic response of the Livermore basin, a major structural depression in the California Coast Range Province bounded by active faults. Site response is calculated by two methods: the reference‐site spectral ratio method and a source‐site spectral inversion method. Longer‐period (≥1  s) amplification factors follow the same general pattern as Bouguer gravity anomaly contours. Site response spectra are inverted for shallow shear‐wave velocity profiles, which are consistent with independent information. Frequency–wavenumber analysis is used to analyze plane‐wave propagation across the Livermore Valley and to identify basin‐edge‐induced surface waves with back azimuths different from the source back azimuth. Finite‐element simulations in a 3D velocity model of the region illustrate the generation of basin‐edge‐induced surface waves and point out strips of elevated ground velocities along the margins of the basin.

Insights into the Structure of Intrastrand Cross-Link DNA Lesion-Containing Oligonucleotides: G[8-5m]T and G[8-5]C from Molecular Dynamics Simulations

Czech Academy of Sciences Publication Activity Database

Dumont, E.; Dršata, Tomáš; Guerra, C. F.; Lankaš, Filip

2015-01-01

Roč. 54, č. 5 (2015), s. 1259-1267 ISSN 0006-2960 R&D Projects: GA ČR(CZ) GA14-21893S Institutional support: RVO:61388963 Keywords : hydrogen bonds * abasic sites * duplex DNA Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.876, year: 2015

Development and evaluation of human AP endonuclease inhibitors in melanoma and glioma cell lines

DEFF Research Database (Denmark)

Mohammed, M Z; Vyjayanti, V N; Laughton, C A

2011-01-01

Modulation of DNA base excision repair (BER) has the potential to enhance response to chemotherapy and improve outcomes in tumours such as melanoma and glioma. APE1, a critical protein in BER that processes potentially cytotoxic abasic sites (AP sites), is a promising new target in cancer. In the....... In the current study, we aimed to develop small molecule inhibitors of APE1 for cancer therapy....

Update History of This Database - KOME | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available List Contact us KOME Update History of This Database Date Update contents 2014/10/22 The URL of the whole da...site is opened. 2003/07/18 KOME ( http://cdna01.dna.affrc.go.jp/cDNA/ ) is opened. About This Database Dat...abase Description Download License Update History of This Database Site Policy | Contact Us Update History of This Database - KOME | LSDB Archive ...

Assessment of earthquake-induced tsunami hazard at a power plant site

International Nuclear Information System (INIS)

Ghosh, A.K.

2008-01-01

This paper presents a study of the tsunami hazard due to submarine earthquakes at a power plant site on the east coast of India. The paper considers various sources of earthquakes from the tectonic information, and records of past earthquakes and tsunamis. Magnitude-frequency relationship for earthquake occurrence rate and a simplified model for tsunami run-up height as a function of earthquake magnitude and the distance between the source and site have been developed. Finally, considering equal likelihood of generation of earthquakes anywhere on each of the faults, the tsunami hazard has been evaluated and presented as a relationship between tsunami height and its mean recurrence interval (MRI). Probability of exceedence of a certain wave height in a given period of time is also presented. These studies will be helpful in making an estimate of the tsunami-induced flooding potential at the site

The role of HERC2 and RNF8 ubiquitin E3 ligases in the promotion of translesion DNA synthesis in the chicken DT40 cell line

DEFF Research Database (Denmark)

Mohiuddin, Mohammed; Kobayashi, Shunsuke; Keka, Islam Shamima

2016-01-01

immediately after exposure to UV while retaining proficient post-replicative gap filling. These mutants are both proficient in mono-ubiquitination of PCNA. Taken together, these results suggest that HERC2 and RNF8 promote TLS past abasic sites and UV-lesions at or very close to stalled replication forks....

Enoxaparin-induced skin necrosis at injection site after total knee arthroplasty

Directory of Open Access Journals (Sweden)

Max Haffner, BS

2018-03-01

Full Text Available Enoxaparin is a widely used low-molecular-weight heparin for perioperative thromboembolic prophylaxis. Enoxaparin-induced skin necrosis in the setting of arthroplasty has been rarely reported in the literature with varying outcomes and management decisions. Our patient developed skin necrosis at his injection site and thrombocytopenia 10 days following left total knee arthroplasty surgery and after receiving subcutaneous Lovenox injections postoperatively. The patient was started on an alternative anticoagulation based on a high suspicion for heparin-induced thrombocytopenia and the wound was monitored without surgical debridement. Our case highlights the key clinical management decisions when facing this potentially life-threatening adverse reaction. Keywords: Lovenox, Enoxaparin, Skin necrosis, Adverse reaction, Arthroplasty

pSort search result - KOME | LSDB Archive [Life Science Database Archive metadata

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Full Text Available switchLanguage; BLAST Search Image Search Home About Archive Update History Data ...name: kome_psort_search_result.zip File URL: ftp://ftp.biosciencedbc.jp/archive/kome/LATEST/kome_psort_searc...abase Description Download License Update History of This Database Site Policy | Contact Us pSort search result - KOME | LSDB Archive ...

External main-induced events in relation to nuclear power plant siting

International Nuclear Information System (INIS)

1981-01-01

This safety Guide recomments procedures and provides information for use in implementing that part of the code of safety in Nuclear Power Plant Siting (IAEA Safety Series No. 50-C-S) which concerns man-induced events external to the plant, up to the evaluation of corresponding design basis parameters. Like the code, the Guide forms part of the IAEA's programme, referred to as the NUSS programme, for establishing codes of practice and safety Guides relating to land-based stationary thermal neutron power plants

Database Description - PSCDB | LSDB Archive [Life Science Database Archive metadata

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Full Text Available abase Description General information of database Database name PSCDB Alternative n...rial Science and Technology (AIST) Takayuki Amemiya E-mail: Database classification Structure Databases - Protein structure Database...554-D558. External Links: Original website information Database maintenance site Graduate School of Informat...available URL of Web services - Need for user registration Not available About This Database Database Descri...ption Download License Update History of This Database Site Policy | Contact Us Database Description - PSCDB | LSDB Archive ...

Phloem development in nematode-induced feeding sites: The implications of auxin and cytokinin

Directory of Open Access Journals (Sweden)

Birgit eAbsmanner

2013-07-01

Full Text Available Sedentary plant parasitic nematodes such as root-knot nematodes and cyst nematodes induce giant cells or syncytia, respectively, in their host plant’s roots. These highly specialized structures serve as feeding sites from which exclusively the nematodes withdraw nutrients. While giant cells are symplastically isolated and obtain assimilates by transporter-mediated processes syncytia are massively connected to the phloem by plasmodesmata. To support the feeding sites and the nematode during their development, phloem is induced around syncytia and giant cells. In the case of syncytia the unloading phloem consists of sieve elements and companion cells and in the case of root knots it consists exclusively of sieve elements. We applied immunohistochemistry to identify the cells within the developing phloem that responded to auxin and cytokinin. Both feeding sites themselves did not respond to either hormone. We were able to show that in root knots an auxin response precedes the differentiation of these auxin responsive cells into phloem elements. This process appears to be independent of B-type Arabidopsis response regulators. Using additional markers for tissue identity we provide evidence that around giant cells protophloem is formed and proliferates dramatically. In contrast, the phloem around syncytia responded to both hormones. The presence of companion cells as well as hormone-responsive sieve elements suggests that metaphloem development occurs. The implication of auxin and cytokinin in the further development of the metaphloem is discussed.

Update History of This Database - KAIKOcDNA | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available switchLanguage; BLAST Search Image Search Home About Archive Update History Data ...List Contact us KAIKOcDNA Update History of This Database Date Update contents 2014/10/20 The URL of the dat... database ( http://sgp.dna.affrc.go.jp/EST/ ) is opened. About This Database Database Description Download License Update Hi...story of This Database Site Policy | Contact Us Update History of This Database - KAIKOcDNA | LSDB Archive ... ...abase maintenance site is changed. 2014/10/08 KAIKOcDNA English archive site is opened. 2004/04/12 KAIKOcDNA

Interstrand cross-linking implies contrasting structural consequences for DNA: insights from molecular dynamics

Czech Academy of Sciences Publication Activity Database

Bignon, E.; Dršata, Tomáš; Morell, C.; Lankaš, Filip; Dumont, E.

2017-01-01

Roč. 45, č. 4 (2017), s. 2188-2195 ISSN 0305-1048 R&D Projects: GA ČR(CZ) GA14-21893S Institutional support: RVO:61388963 Keywords : abasic sites * duplex DNA * mechanical properties Subject RIV: BO - Biophysics OBOR OECD: Biophysics Impact factor: 10.162, year: 2016 https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/gkw1253

Influence of the complexity of radiation-induced DNA damage on enzyme recognition

International Nuclear Information System (INIS)

Palmer, Philip

2002-01-01

Ionising radiation is unique in inducing DNA clustered damage together with the simple isolated lesions. Understanding how these complex lesions are recognised and repaired by the cell is key to understanding the health risks associated with radiation exposure. This study focuses on whether ionising radiation-induced complex single-strand breaks (SSB) are recognised by DNA-PK and PARP, and whether the complexity of DSB influence their ligation by either DNA ligase lV/XRCC4 (LX) complex or T4 DNA ligase. Plasmid DNA, irradiated in aqueous solution using sparsely ionising γ-rays and densely ionising α-particles produce different yields of complex DNA damages, used as substrates for in vitro DNA-PK and PARP activity assays. The activity of DNA-PK to phosphorylate a peptide was determined using HF19 cell nuclear extracts as a source of DNA-PK. PARP ADP-ribosylation activity was determined using purified PARP enzyme. The activation of DNA-PK and PARP by irradiated DNA is due to SSB and not the low yield of DSB (linear plasmid DNA <10%). A ∼2 fold increase in DNA-PK activation and a ∼3-fold reduction in PARP activity seen on increasing the ionising density of the radiation (proportion of complex damage) are proposed to reflect changes in the complexity of SSB and may relate to damage signalling. Complex DSB synthesised as double-stranded oligonucleotides, with a 2 bp 5'-overhang, and containing modified lesions, 8-oxoguanine and abasic sites, at known positions relative to the termini were used as substrates for in vitro ligation by DNA ligase IV/XRCC4 or T4 ligase. The presence of a modified lesion 2 or 3 bp but not 4 bp from the 3'-termini and 2 or 6 bp from the 5'-termini caused a drastic reduction in the extent of ligation. Therefore, the presence of modified lesions near to the termini of a DSB may compromise their rejoining by non-homologous end-joining (NHEJ) involving the LX complex. (author)

Database Description - AcEST | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available abase Description General information of database Database name AcEST Alternative n...hi, Tokyo-to 192-0397 Tel: +81-42-677-1111(ext.3654) E-mail: Database classificat...eneris Taxonomy ID: 13818 Database description This is a database of EST sequences of Adiantum capillus-vene...(3): 223-227. External Links: Original website information Database maintenance site Plant Environmental Res...base Database Description Download License Update History of This Database Site Policy | Contact Us Database Description - AcEST | LSDB Archive ...

Increased 3H-spiperone binding sites in mesolimbic area related to methamphetamine-induced behavioral hypersensitivity

International Nuclear Information System (INIS)

Akiyama, K.; Sato, M.; Otsuki, S.

1982-01-01

The specific 3 H-spiperone binding to membrane homogenates of the striatum, mesolimbic area, and frontal cortex was examined in two groups of rats pretreated once daily with saline or 4 mg/kg of methamphetamine (MAP) for 14 days. At 7 days following cessation of chronic pretreatment, all rats received an injection of 4 mg/kg of MAP and were decapitated 1 hr after the injection. In the chronic saline-pretreatment group, the single administration of MAP induced significant changes in the number (Bmax) of specific 3 H-spiperone binding sites (a decrease in the striatum and an increase in the mesolimbic area and frontal cortex), but no significant changes in the affinity (KD) in any brain area. The chronic MAP pretreatment markedly augmented the changes in Bmax in the striatum and mesolimbic area. The increase in specific 3 H-spiperone binding sites in the mesolimbic area is discussed in relation to MAP-induced behavioral hypersensitivity

On-bead fluorescent DNA nanoprobes to analyze base excision repair activities

International Nuclear Information System (INIS)

Gines, Guillaume; Saint-Pierre, Christine; Gasparutto, Didier

2014-01-01

Graphical abstract: -- Highlights: •On magnetic beads fluorescent enzymatic assays. •Simple, easy, non-radioactive and electrophoresis-free functional assay. •Lesion-containing hairpin DNA probes are selective for repair enzymes. •The biosensing platform allows the measurement of DNA repair activities from purified enzymes or within cell free extracts. -- Abstract: DNA integrity is constantly threatened by endogenous and exogenous agents that can modify its physical and chemical structure. Changes in DNA sequence can cause mutations sparked by some genetic diseases or cancers. Organisms have developed efficient defense mechanisms able to specifically repair each kind of lesion (alkylation, oxidation, single or double strand break, mismatch, etc). Here we report the adjustment of an original assay to detect enzymes’ activity of base excision repair (BER), that supports a set of lesions including abasic sites, alkylation, oxidation or deamination products of bases. The biosensor is characterized by a set of fluorescent hairpin-shaped nucleic acid probes supported on magnetic beads, each containing a selective lesion targeting a specific BER enzyme. We have studied the DNA glycosylase alkyl-adenine glycosylase (AAG) and the human AP-endonuclease (APE1) by incorporating within the DNA probe a hypoxanthine lesion or an abasic site analog (tetrahydrofuran), respectively. Enzymatic repair activity induces the formation of a nick in the damaged strand, leading to probe's break, that is detected in the supernatant by fluorescence. The functional assay allows the measurement of DNA repair activities from purified enzymes or in cell-free extracts in a fast, specific, quantitative and sensitive way, using only 1 pmol of probe for a test. We recorded a detection limit of 1 μg mL −1 and 50 μg mL −1 of HeLa nuclear extracts for APE1 and AAG enzymes, respectively. Finally, the on-bead assay should be useful to screen inhibitors of DNA repair activities

Tissue characteristics of high- and low-incidence plutonium-induced osteogenic sarcoma sites in life-span beagles

International Nuclear Information System (INIS)

Miller, S.C.; Jee, W.S.S.; Smith, J.M.; Wronski, T.J.

1986-01-01

On the basis of information gathered from the 239 Pu life-span study in beagles at the University of Utah, the tissue features were found to be characteristic of high-incidence bone-tumor sites compared to low-incidence sites included more hematopoietic tissues in the bone marrow; greater trabecular bone mass; greater bone remodeling rates; greater mineral apposition rates; greater density and activity of bone surface cells; greater density of putative bone-cell precursors; greater initial uptake of plutonium on bone surfaces; and greater marrow vascular volumes and a venous sinusoidal bed. Although most of these studies are not yet complete, the information being collected should contribute to our understanding of the mechanisms of radiation-induced osteogenic sarcomas. This should aid in predicting the types and characteristics of osseous tissues where radiation-induced osteogenic sarcomas may arise in humans. 25 refs., 4 figs., 3 tabs

Database Description - ASTRA | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available abase Description General information of database Database name ASTRA Alternative n...tics Journal Search: Contact address Database classification Nucleotide Sequence Databases - Gene structure,...3702 Taxonomy Name: Oryza sativa Taxonomy ID: 4530 Database description The database represents classified p...(10):1211-6. External Links: Original website information Database maintenance site National Institute of Ad... for user registration Not available About This Database Database Description Dow

2,3,7,8-Tetrachlorodibenzo-p-dioxin modulates estradiol-induced aldehydic DNA lesions in human breast cancer cells through alteration of CYP1A1 and CYP1B1 expression.

Science.gov (United States)

Chen, Shou-Tung; Chen, Dar-Ren; Fang, Ju-Pin; Lin, Po-Hsiung

2015-05-01

Many genes responsible for the bioactivation of endogenous estrogen to reactive quinonoid metabolites, including cytochrome P450 (CYP) 1A1, 1A2, and 1B1, are well-known target genes of the aryl hydrocarbon receptor agonist 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The purpose of this research was to investigate the roles of TCDD-mediated altered gene expression in the induction of aldehydic DNA lesions (ADLs) by 17β-estradiol (E2) in human MDA-MB-231 and MCF-7 breast cancer cells. We demonstrated that increases in the number of oxidant-mediated ADLs, including abasic sites and aldehydic base/sugar lesions, were detected in MDA-MB-231 cells exposed to E2. The DNA-damaging effects of E2 in MDA-MB-231 cells were prevented by pretreatment of cells with TCDD. In contrast, we did not observe statistically significant increases in the number of ADLs in MCF-7 cells exposed to E2. However, with TCDD pretreatment, an approximately twofold increase in the number of ADLs was detected in MCF-7 cells exposed to E2. TCDD pretreatment induces disparity in the disposition of E2 to reactive quinonoid metabolites and the subsequent formation of oxidative DNA lesions through alteration of CYP1A1 and CYP1B1 expression in human breast cancer cells.

The formation of double-strand breaks at multiply damaged sites is driven by the kinetics of excision/incision at base damage in eukaryotic cells

International Nuclear Information System (INIS)

Kozmin, S.G.; Sedletska, Y.; Reynaud-Angelin, A.; Sage, E.; Kozmin, S.G.; Sedletska, Y.; Reynaud-Angelin, A.; Sage, E.; Gasparutto, D.

2009-01-01

It has been stipulated that repair of clustered DNA lesions may be compromised, possibly leading to the formation of double-strand breaks (DSB) and, thus, to deleterious events. Using a variety of model multiply damaged sites (MDS), we investigated parameters that govern the formation of DSB during the processing of MDS. Duplexes carrying MDS were inserted into replicative or integrative vectors, and used to transform yeast Saccharomyces cerevisiae. Formation of DSB was assessed by a relevant plasmid survival assay. Kinetics of excision/incision and DSB formation at MDS was explored using yeast cell extracts. We show that MDS composed of two uracils or abasic sites, were rapidly incised and readily converted into DSB in yeast cells. In marked contrast, none of the MDS carrying opposed oG and hU separated by 38 bp gave rise to DSB, despite the fact that some of them contained preexisting single-strand break (a 1-nt gap). Interestingly, the absence of DSB formation in this case correlated with slow excision/incision rates of lesions. We propose that the kinetics of the initial repair steps at MDS is a major parameter that direct towards the conversion of MDS into DSB. Data provides clues to the biological consequences of MDS in eukaryotic cells. (authors)

Evidence for a high and a low sensitivity receptor site for radiation-induced vomiting in the dog

International Nuclear Information System (INIS)

Harding, R.K.; Hugenholtz, H.; Kucharczyk, J.

1987-01-01

Ablation of the area postrema (AP) has been shown to prevent radiation-induced vomiting in the dog (induced by 6-8 By /sup 60/Co γ). In other species (cat, monkey) a higher dose of radiation is necessary to elicit emesis and visceral deafferentation has been more effective than AP ablation in preventing vomiting. The AP and the dorsal motor nucleus of the vagus (DMV), an important visceral afferent terminus, are anatomically adjacent structures in the medulla oblongata. Aggressive AP ablation could affect the DMV. The author's removed the AP in 6 dogs. Previous work showed this surgery prevented vomiting following exposure to 6-8 Gy. To prove the lesion did not invade the underlying DMV, the animals were given a hypertonic saline gavage, to elicit vomiting via visceral receptors. Animals were later submitted to 15-20 Gy. Those with proved visceral afferent connections vomited. They conclude that the AP is the most sensitive site for the stimulation of radiation-induced vomiting and that a less sensitive visceral site also exists. Differences in the relative sensitivities of these 2 systems may account for published species differences

Ca-site substitution induced a metal-insulator transition in manganite CaMnO3

International Nuclear Information System (INIS)

Sousa, D.; Nunes, M.R.; Silveira, C.; Matos, I.; Lopes, A.B.; Melo Jorge, M.E.

2008-01-01

A systematic study of the A-site doping in Mn(IV)-rich perovskite manganites Ca 1-x Ho x MnO 3 , over a large homogeneity range (0.1 ≤ x ≤ 0.4), has been performed. A significant increase in the lattice parameters indicated the presence of mixed valence state of Mn: Mn 3+ and Mn 4+ . The substitution of calcium by holmium also induces strong changes in the electrical properties. We found that small Ho concentration produces an important decrease in the electrical resistivity and induces an electrical transition, the temperature corresponding to the metal-insulator transition (T MI ) shifts with the holmium content. This electrical behavior is attributed to the Mn 3+ ions content and a charge order effect

Radiation affects binding of Fpg repair protein to an abasic site containing DNA

Czech Academy of Sciences Publication Activity Database

Gillard, N.; Běgusová, Marie; Castaing, B.; Spotheim-Maurizot, M.

2004-01-01

Roč. 162, č. 5 (2004), s. 566-571 ISSN 0033-7587 R&D Projects: GA AV ČR IAA1048103 Institutional research plan: CEZ:AV0Z1048901 Keywords : ionizing radiation * DNA * protein komplex Subject RIV: BO - Biophysics Impact factor: 3.208, year: 2003

Adaptation to Alkylation Damage in DNA Measured by the comet Assay

Czech Academy of Sciences Publication Activity Database

Angelis, Karel; McGuffie, M.; Menke, M.; Schubert, I.

2000-01-01

Roč. 36, - (2000), s. 146-150 ISSN 0893-6692 R&D Projects: GA ČR GA203/98/0682; GA ČR GA204/97/0154 Grant - others:-(XC) P2020702; -(DE) Schu 951/5-1; a-(DE) 436TSE17/7/98 Institutional research plan: CEZ:AV0Z5038910 Keywords : abasic (AP) sites * AP-endonuclease Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.278, year: 2000

Database Description - ConfC | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available abase Description General information of database Database name ConfC Alternative name Database...amotsu Noguchi Tel: 042-495-8736 E-mail: Database classification Structure Database...s - Protein structure Structure Databases - Small molecules Structure Databases - Nucleic acid structure Database... services - Need for user registration - About This Database Database Description Download License Update History of This Database... Site Policy | Contact Us Database Description - ConfC | LSDB Archive ...

Database Description - PLACE | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available abase Description General information of database Database name PLACE Alternative name A Database...Kannondai, Tsukuba, Ibaraki 305-8602, Japan National Institute of Agrobiological Sciences E-mail : Databas...e classification Plant databases Organism Taxonomy Name: Tracheophyta Taxonomy ID: 58023 Database...99, Vol.27, No.1 :297-300 External Links: Original website information Database maintenance site National In...- Need for user registration Not available About This Database Database Descripti

Background studies: human-induced effects on the evolution of shallow land burial sites for radioactive waste disposal

International Nuclear Information System (INIS)

1987-11-01

This report presents the results of a programme of background research on the human-induced effects on the long term evolution of shallow disposal sites for low level radioactive wastes. The work is intended to support development and use of the TIME2 simulation code. Within the context of climatic change up to the next glacial maximum three areas are addressed: planning and legislative control over site usage, biosphere state changes and intrusion. An appendix presents a discussion of some planning aspects of radioactive waste disposal. (author)

Molecular characterization of a complex site-specific radiation-induced DNA double-strand break

International Nuclear Information System (INIS)

Datta, K.; Dizdaroglu, M.; Jaruga, P.; Neumann, R.D.; Winters, T.A.

2003-01-01

Radiation lethality is a function of radiation-induced DNA double-strand breaks (DSB). Current models propose the lethality of a DSB to be a function of its structural complexity. We present here for the first time a map of damage associated with a site-specific double-strand break produced by decay of 125 I in a plasmid bound by a 125 I-labeled triplex forming oligonucleotide ( 125 I-TFO). The E. coli DNA repair enzymes, endonuclease IV (endo IV), endonuclease III (endo III), and formamidopyrimidine-DNA glycosylase (Fpg), which recognize AP sites, and pyrimidine and purine base damage respectively, were used as probes in this study. 125 I-TFO bound plasmid was incubated with and without DMSO at -80 deg C for 1 month. No significant difference in DSB yield was observed under these conditions. A 32 base pair fragment from the upstream side of the decay site was isolated by restriction digestion and enzymatically probed to identify damage sites. Endo IV treatment of the 5'-end labeled upper strand indicated clustering of AP sites within 3 bases downstream and 7 bases upstream of the targeted base. Also, repeated experiments consistently detected an AP site 4 bases upstream of the 125 Itarget base. This was further supported by complementary results with the 3'-end labeled upper strand. Endo IV analysis of the lower strand also shows clustering of AP sites near the DSB end. Endo III and Fpg probing demonstrated that base damage is also clustered near the targeted break site. DSBs produced in the absence of DMSO displayed a different pattern of enzyme sensitive damage than those produced in the presence of DMSO. Identification of specific base damage types within the restriction fragment containing the DSB end was achieved with GC/MS. Base damage consisted of 8-hydroguanine, 8-hydroxyadenine, and 5-hydroxycytosine. These lesions were observed at relative yields of 8-hydroguanine and 5-hydroxycytosine to 8-hydroxyadenine of 7.4:1 and 4.7:1, respectively, in the absence

Full-Length Sequence of Mouse Acupuncture-Induced 1-L (Aig1l Gene Including Its Transcriptional Start Site

Directory of Open Access Journals (Sweden)

Mika Ohta

2011-01-01

Full Text Available We have been investigating the molecular efficacy of electroacupuncture (EA, which is one type of acupuncture therapy. In our previous molecular biological study of acupuncture, we found an EA-induced gene, named acupuncture-induced 1-L (Aig1l, in mouse skeletal muscle. The aims of this study consisted of identification of the full-length cDNA sequence of Aig1l including the transcriptional start site, determination of the tissue distribution of Aig1l and analysis of the effect of EA on Aig1l gene expression. We determined the complete cDNA sequence including the transcriptional start site via cDNA cloning with the cap site hunting method. We then analyzed the tissue distribution of Aig1l by means of northern blot analysis and real-time quantitative polymerase chain reaction. We used the semiquantitative reverse transcriptase-polymerase chain reaction to examine the effect of EA on Aig1l gene expression. Our results showed that the complete cDNA sequence of Aig1l was 6073 bp long, and the putative protein consisted of 962 amino acids. All seven tissues that we analyzed expressed the Aig1l gene. In skeletal muscle, EA induced expression of the Aig1l gene, with high expression observed after 3 hours of EA. Our findings thus suggest that the Aig1l gene may play a key role in the molecular mechanisms of EA efficacy.

Artificial neural network for on-site quantitative analysis of soils using laser induced breakdown spectroscopy

Energy Technology Data Exchange (ETDEWEB)

El Haddad, J. [Univ. Bordeaux, LOMA, UMR 5798, F-33400 Talence (France); CNRS, LOMA, UMR 5798, F-33400 Talence (France); Villot-Kadri, M.; Ismaël, A.; Gallou, G. [IVEA Solution, Centre Scientifique d' Orsay, Bât 503, 91400 Orsay (France); Michel, K.; Bruyère, D.; Laperche, V. [BRGM, Service Métrologie, Monitoring et Analyse, 3 avenue Claude Guillemin, B.P 36009, 45060 Orléans Cedex (France); Canioni, L. [Univ. Bordeaux, LOMA, UMR 5798, F-33400 Talence (France); CNRS, LOMA, UMR 5798, F-33400 Talence (France); Bousquet, B., E-mail: bruno.bousquet@u-bordeaux1.fr [Univ. Bordeaux, LOMA, UMR 5798, F-33400 Talence (France); CNRS, LOMA, UMR 5798, F-33400 Talence (France)

2013-01-01

Nowadays, due to environmental concerns, fast on-site quantitative analyses of soils are required. Laser induced breakdown spectroscopy is a serious candidate to address this challenge and is especially well suited for multi-elemental analysis of heavy metals. However, saturation and matrix effects prevent from a simple treatment of the LIBS data, namely through a regular calibration curve. This paper details the limits of this approach and consequently emphasizes the advantage of using artificial neural networks well suited for non-linear and multi-variate calibration. This advanced method of data analysis is evaluated in the case of real soil samples and on-site LIBS measurements. The selection of the LIBS data as input data of the network is particularly detailed and finally, resulting errors of prediction lower than 20% for aluminum, calcium, copper and iron demonstrate the good efficiency of the artificial neural networks for on-site quantitative LIBS of soils. - Highlights: ► We perform on-site quantitative LIBS analysis of soil samples. ► We demonstrate that univariate analysis is not convenient. ► We exploit artificial neural networks for LIBS analysis. ► Spectral lines other than the ones from the analyte must be introduced.

Artificial neural network for on-site quantitative analysis of soils using laser induced breakdown spectroscopy

International Nuclear Information System (INIS)

El Haddad, J.; Villot-Kadri, M.; Ismaël, A.; Gallou, G.; Michel, K.; Bruyère, D.; Laperche, V.; Canioni, L.; Bousquet, B.

2013-01-01

Nowadays, due to environmental concerns, fast on-site quantitative analyses of soils are required. Laser induced breakdown spectroscopy is a serious candidate to address this challenge and is especially well suited for multi-elemental analysis of heavy metals. However, saturation and matrix effects prevent from a simple treatment of the LIBS data, namely through a regular calibration curve. This paper details the limits of this approach and consequently emphasizes the advantage of using artificial neural networks well suited for non-linear and multi-variate calibration. This advanced method of data analysis is evaluated in the case of real soil samples and on-site LIBS measurements. The selection of the LIBS data as input data of the network is particularly detailed and finally, resulting errors of prediction lower than 20% for aluminum, calcium, copper and iron demonstrate the good efficiency of the artificial neural networks for on-site quantitative LIBS of soils. - Highlights: ► We perform on-site quantitative LIBS analysis of soil samples. ► We demonstrate that univariate analysis is not convenient. ► We exploit artificial neural networks for LIBS analysis. ► Spectral lines other than the ones from the analyte must be introduced

Radiolysis of DNA-protein complexes

Energy Technology Data Exchange (ETDEWEB)

Begusova, Marie [Department of Radiation Dosimetry, Nuclear Physics Institute, Na Truhlarce 39/64, CZ-18086, Prague 8 (Czech Republic)]. E-mail: begusova@ujf.cas.cz; Gillard, Nathalie [Centre de Biophysique Moleculaire, CNRS, rue Charles-Sadron, F-45071 Orleans Cedex 2 (France); Sy, Denise [Centre de Biophysique Moleculaire, CNRS, rue Charles-Sadron, F-45071 Orleans Cedex 2 (France); Castaing, Bertrand [Centre de Biophysique Moleculaire, CNRS, rue Charles-Sadron, F-45071 Orleans Cedex 2 (France); Charlier, Michel [Centre de Biophysique Moleculaire, CNRS, rue Charles-Sadron, F-45071 Orleans Cedex 2 (France); Spotheim-Maurizot, Melanie [Centre de Biophysique Moleculaire, CNRS, rue Charles-Sadron, F-45071 Orleans Cedex 2 (France)

2005-02-01

We discuss here modifications of DNA and protein radiolysis due to the interaction of these two partners in specific complexes. Experimental patterns of frank strand breaks (FSB) and alkali revealed breaks (ARB) obtained for DNA lac operator bound to the lac repressor and for a DNA containing an abasic site analog bound to the formamidopyrimidine-DNA glycosylase are reported. Experimental data are compared to predicted damage distribution obtained using the theoretical model RADACK.

Radiolysis of DNA-protein complexes

International Nuclear Information System (INIS)

Begusova, Marie; Gillard, Nathalie; Sy, Denise; Castaing, Bertrand; Charlier, Michel; Spotheim-Maurizot, Melanie

2005-01-01

We discuss here modifications of DNA and protein radiolysis due to the interaction of these two partners in specific complexes. Experimental patterns of frank strand breaks (FSB) and alkali revealed breaks (ARB) obtained for DNA lac operator bound to the lac repressor and for a DNA containing an abasic site analog bound to the formamidopyrimidine-DNA glycosylase are reported. Experimental data are compared to predicted damage distribution obtained using the theoretical model RADACK

Database Description - GETDB | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available abase Description General information of database Database name GETDB Alternative n...ame Gal4 Enhancer Trap Insertion Database DOI 10.18908/lsdba.nbdc00236-000 Creator Creator Name: Shigeo Haya... Chuo-ku, Kobe 650-0047 Tel: +81-78-306-3185 FAX: +81-78-306-3183 E-mail: Database classification Expression... Invertebrate genome database Organism Taxonomy Name: Drosophila melanogaster Taxonomy ID: 7227 Database des...riginal website information Database maintenance site Drosophila Genetic Resource

Tolerance to LSD and DOB induced shaking behaviour: differential adaptations of frontocortical 5-HT(2A) and glutamate receptor binding sites.

Science.gov (United States)

Buchborn, Tobias; Schröder, Helmut; Dieterich, Daniela C; Grecksch, Gisela; Höllt, Volker

2015-03-15

Serotonergic hallucinogens, such as lysergic acid diethylamide (LSD) and dimethoxy-bromoamphetamine (DOB), provoke stereotype-like shaking behaviour in rodents, which is hypothesised to engage frontocortical glutamate receptor activation secondary to serotonin2A (5-HT2A) related glutamate release. Challenging this hypothesis, we here investigate whether tolerance to LSD and DOB correlates with frontocortical adaptations of 5-HT2A and/or overall-glutamate binding sites. LSD and DOB (0.025 and 0.25 mg/kg, i.p.) induce a ketanserin-sensitive (0.5 mg/kg, i.p., 30-min pretreatment) increase in shaking behaviour (including head twitches and wet dog shakes), which with repeated application (7× in 4 ds) is undermined by tolerance. Tolerance to DOB, as indexed by DOB-sensitive [(3)H]spiroperidol and DOB induced [(35)S]GTP-gamma-S binding, is accompanied by a frontocortical decrease in 5-HT2A binding sites and 5-HT2 signalling, respectively; glutamate-sensitive [(3)H]glutamate binding sites, in contrast, remain unchanged. As to LSD, 5-HT2 signalling and 5-HT2A binding, respectively, are not or only marginally affected, yet [(3)H]glutamate binding is significantly decreased. Correlation analysis interrelates tolerance to DOB to the reduced 5-HT2A (r=.80) as well as the unchanged [(3)H]glutamate binding sites (r=.84); tolerance to LSD, as opposed, shares variance with the reduction in [(3)H]glutamate binding sites only (r=.86). Given that DOB and LSD both induce tolerance, one correlating with 5-HT2A, the other with glutamate receptor adaptations, it might be inferred that tolerance can arise at either level. That is, if a hallucinogen (like LSD in our study) fails to induce 5-HT2A (down-)regulation, glutamate receptors (activated postsynaptic to 5-HT2A related glutamate release) might instead adapt and thus prevent further overstimulation of the cortex. Copyright © 2014 Elsevier B.V. All rights reserved.

Long-Range Electrostatics-Induced Two-Proton Transfer Captured by Neutron Crystallography in an Enzyme Catalytic Site.

Science.gov (United States)

Gerlits, Oksana; Wymore, Troy; Das, Amit; Shen, Chen-Hsiang; Parks, Jerry M; Smith, Jeremy C; Weiss, Kevin L; Keen, David A; Blakeley, Matthew P; Louis, John M; Langan, Paul; Weber, Irene T; Kovalevsky, Andrey

2016-04-11

Neutron crystallography was used to directly locate two protons before and after a pH-induced two-proton transfer between catalytic aspartic acid residues and the hydroxy group of the bound clinical drug darunavir, located in the catalytic site of enzyme HIV-1 protease. The two-proton transfer is triggered by electrostatic effects arising from protonation state changes of surface residues far from the active site. The mechanism and pH effect are supported by quantum mechanics/molecular mechanics (QM/MM) calculations. The low-pH proton configuration in the catalytic site is deemed critical for the catalytic action of this enzyme and may apply more generally to other aspartic proteases. Neutrons therefore represent a superb probe to obtain structural details for proton transfer reactions in biological systems at a truly atomic level. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

On-bead fluorescent DNA nanoprobes to analyze base excision repair activities

Energy Technology Data Exchange (ETDEWEB)

Gines, Guillaume; Saint-Pierre, Christine; Gasparutto, Didier, E-mail: didier.gasparutto@cea.fr

2014-02-17

Graphical abstract: -- Highlights: •On magnetic beads fluorescent enzymatic assays. •Simple, easy, non-radioactive and electrophoresis-free functional assay. •Lesion-containing hairpin DNA probes are selective for repair enzymes. •The biosensing platform allows the measurement of DNA repair activities from purified enzymes or within cell free extracts. -- Abstract: DNA integrity is constantly threatened by endogenous and exogenous agents that can modify its physical and chemical structure. Changes in DNA sequence can cause mutations sparked by some genetic diseases or cancers. Organisms have developed efficient defense mechanisms able to specifically repair each kind of lesion (alkylation, oxidation, single or double strand break, mismatch, etc). Here we report the adjustment of an original assay to detect enzymes’ activity of base excision repair (BER), that supports a set of lesions including abasic sites, alkylation, oxidation or deamination products of bases. The biosensor is characterized by a set of fluorescent hairpin-shaped nucleic acid probes supported on magnetic beads, each containing a selective lesion targeting a specific BER enzyme. We have studied the DNA glycosylase alkyl-adenine glycosylase (AAG) and the human AP-endonuclease (APE1) by incorporating within the DNA probe a hypoxanthine lesion or an abasic site analog (tetrahydrofuran), respectively. Enzymatic repair activity induces the formation of a nick in the damaged strand, leading to probe's break, that is detected in the supernatant by fluorescence. The functional assay allows the measurement of DNA repair activities from purified enzymes or in cell-free extracts in a fast, specific, quantitative and sensitive way, using only 1 pmol of probe for a test. We recorded a detection limit of 1 μg mL{sup −1} and 50 μg mL{sup −1} of HeLa nuclear extracts for APE1 and AAG enzymes, respectively. Finally, the on-bead assay should be useful to screen inhibitors of DNA repair

Site-specific analysis of UV-induced cyclobutane pyrimidine dimers in nucleotide excision repair-proficient and -deficient hamster cells: Lack of correlation with mutational spectra

International Nuclear Information System (INIS)

Vreeswijk, Maaike P.G.; Meijers, Caro M.; Giphart-Gassler, Micheline; Vrieling, Harry; Zeeland, Albert A. van; Mullenders, Leon H.F.; Loenen, Wil A.M.

2009-01-01

Irradiation of cells with UVC light induces two types of mutagenic DNA photoproducts, i.e. cyclobutane pyrimidine dimers (CPD) and pyrimidine (6-4) pyrimidone photoproducts (6-4PP). To investigate the relationship between the frequency of UV-induced photolesions at specific sites and their ability to induce mutations, we quantified CPD formation at the nucleotide level along exons 3 and 8 of the hprt gene using ligation-mediated PCR, and determined the mutational spectrum of 132 UV-induced hprt mutants in the AA8 hamster cell line and of 165 mutants in its nucleotide excision repair-defective derivative UV5. In AA8 cells, transversions predominated with a strong strand bias towards thymine-containing photolesions in the non-transcribed strand. As hamster AA8 cells are proficient in global genome repair of 6-4PP but selectively repair CPD from the transcribed strand of active genes, most mutations probably resulted from erroneous bypass of CPD in the non-transcribed strand. However, the relative incidence of CPD and the positions where mutations most frequently arose do not correlate. In fact some major damage sites hardly gave rise to the formation of mutations. In the repair-defective UV5 cells, mutations were almost exclusively C > T transitions caused by photoproducts at PyC sites in the transcribed strand. Even though CPD were formed at high frequencies at some TT sites in UV5, these photoproducts did not contribute to mutation induction at all. We conclude that, even in the absence of repair, large variations in the level of induction of CPD at different sites throughout the two exons do not correspond to frequencies of mutation induction.

The homing of bone marrow MSCs to non-osseous sites for ectopic bone formation induced by osteoinductive calcium phosphate

Science.gov (United States)

Song, Guodong; Habibovic, Pamela; Bao, Chongyun; Hu, Jing; van Blitterswijk, Clemens A.; Yuan, Huipin; Chen, Wenchuan; Xu, Hockin H.K.

2013-01-01

Osteoinductive biomaterials are promising for bone repair. There is no direct proof that bone marrow mesenchymal stem cells (BMSCs) home to non-osseous sites and participate in ectopic bone formation induced by osteoinductive bioceramics. The objective of this study was to use a sex-mismatched beagle dog model to investigate BMSC homing via blood circulation to participate in ectopic bone formation via osteoinductive biomaterial. BMSCs of male dogs were injected into female femoral marrow cavity. The survival and stable chimerism of donor BMSCs in recipients were confirmed with polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH). Biphasic calcium phosphate (BCP) granules were implanted in dorsal muscles of female dogs. Y chromosomes were detected in samples harvested from female dogs which had received male BMSCs. At 4 weeks, cells with Y-chromosomes were distributed in the new bone matrix throughout the BCP granule implant. At 6 weeks, cells with Y chromosomes were present in newly mineralized woven bone. TRAP positive osteoclast-like cells were observed in 4-week implants, and the number of such cells decreased from 4 to 6 weeks. These results show that osteoprogenitors were recruited from bone marrow and homed to ectopic site to serve as a cell source for calcium phosphate-induced bone formation. In conclusion, BMSCs were demonstrated to migrate from bone marrow through blood circulation to non-osseous bioceramic implant site to contribute to ectopic bone formation in a canine model. BCP induced new bone in muscles without growth factor delivery, showing excellent osteoinductivity that could be useful for bone tissue engineering. PMID:23298780

Assessing the induced seismicity by hydraulic fracturing at the Wysin site (Poland)

Science.gov (United States)

Ángel López Comino, José; Cesca, Simone; Kriegerowski, Marius; Heimann, Sebastian; Dahm, Torsten; Mirek, Janusz; Lasocky, Stanislaw

2017-04-01

Induced seismicity related to industrial processes including shale gas and oil exploitation is a current issues that implies enough reasons to be concerned. Hydraulic fracturing usually induces weak events. However, scenarios with larger earthquakes are possible, e.g. if the injected fluids alter friction conditions and trigger the failure of neighbouring faults. This work is focused on a hydrofracking experiment monitored in the framework of the SHEER (SHale gas Exploration and Exploitation induced Risks) EU project at the Wysin site, located in the central-western part of the Peribaltic synclise of Pomerania, Poland. A specific network setup has been installed combining surface installation with three small-scale arrays and a shallow borehole installation. The fracking operations were carried out in June and July 2016 at a depth 4000 m. The monitoring has been operational before, during and after the termination of hydraulic fracturing operations. We apply a recently developed automated full waveform detection algorithm based on the stacking of smooth characteristic function and the identification of high coherence in the signals recorded at different stations. The method was tested with synthetic data and different detector levels yielding values of magnitude of completeness around 0.1. An unsupervised detection catalogue is generated with real data for a time period May-September 2016. We identify strong temporal changes (day/night) of the detection performance. A manual revision of the detected signals reveals that most detections are associated to local and regional seismic signals. Only two events could be assigned to the volume potentially affected by the fracking operations.

The Spleen Is an Ideal Site for Inducing Transplanted Islet Graft Expansion in Mice.

Directory of Open Access Journals (Sweden)

Takeshi Itoh

Full Text Available Alternative islet transplantation sites have the potential to reduce the marginal number of islets required to ameliorate hyperglycemia in recipients with diabetes. Previously, we reported that T cell leukemia homeobox 1 (Tlx1+ stem cells in the spleen effectively regenerated into insulin-producing cells in the pancreas of non-obese diabetic mice with end-stage disease. Thus, we investigated the spleen as a potential alternative islet transplantation site. Streptozotocin-induced diabetic C57BL/6 mice received syngeneic islets into the portal vein (PV, beneath the kidney capsule (KC, or into the spleen (SP. The marginal number of islets by PV, KC, or SP was 200, 100, and 50, respectively. Some plasma inflammatory cytokine levels in the SP group were significantly lower than those of the PV group after receiving a marginal number of islets, indicating reduced inflammation in the SP group. Insulin contents were increased 280 days after islet transplantation compared with those immediately following transplantation (p<0.05. Additionally, Tlx1-related genes, including Rrm2b and Pla2g2d, were up-regulated, which indicates that islet grafts expanded in the spleen. The spleen is an ideal candidate for an alternative islet transplantation site because of the resulting reduced inflammation and expansion of the islet graft.

The homing of bone marrow MSCs to non-osseous sites for ectopic bone formation induced by osteoinductive calcium phosphate.

NARCIS (Netherlands)

Song, G.; Habibovic, Pamela; Bao, Chongyun; Hu, J.; van Blitterswijk, Clemens; Yuan, Huipin; Chen, W.; Xu, H.H.K.

2013-01-01

Osteoinductive biomaterials are promising for bone repair. There is no direct proof that bone marrow mesenchymal stem cells (BMSCs) home to non-osseous sites and participate in ectopic bone formation induced by osteoinductive bioceramics. The objective of this study was to use a sex-mismatched

Induction of Abasic Sites by the Drinking-Water Mutagen MX in Salmonella TA100

Science.gov (United States)

Mutagen X (MX) is a chlorinated furanone that accounts for more of the mutagenic activity of drinking water than any other disinfection by-product. It is one of the most potent base-substitution mutagens in the Salmonella (Ames) mutagenicity assay, producing primarily GC to TA mu...

Benzodiazepines have high-affinity binding sites and induce melanogenesis in B16/C3 melanoma cells.

OpenAIRE

Matthew, E; Laskin, J D; Zimmerman, E A; Weinstein, I B; Hsu, K C; Engelhardt, D L

1981-01-01

We found that two markers of differentiation, tyrosinase (monophenol, dihydroxyphenylalanine:oxygen oxidoreductase, EC 1.14.18.1) activity and melanin synthesis, are induced by diazepam in B16/C3 mouse melanoma cells. We also demonstrated high-affinity binding sites for [3H]diazepam in these cells by radioreceptor assay, and we visualized binding to the cell surface by fluorescence microscopy with a benzodiazepine analog conjugated to a fluorescein-labeled protein. Our studies also showed tha...

Induced Seismicity at the UK "Hot Dry Rock" Test Site for Geothermal Energy Production

Science.gov (United States)

Li, Xun; Main, Ian; Jupe, Andrew

2018-03-01

In enhanced geothermal systems (EGS), fluid is injected at high pressure in order to stimulate fracturing and/or fluid flow through otherwise relatively impermeable underlying hot rocks to generate power and/or heat. The stimulation induces micro-earthquakes whose precise triggering mechanism and relationship to new and pre-existing fracture networks are still the subject of some debate. Here we analyse the dataset for induced micro-earthquakes at the UK "hot dry rock" experimental geothermal site (Rosemanowes, Cornwall). We quantify the evolution of several metrics used to characterise induced seismicity, including the seismic strain partition factor and the "seismogenic index". The results show a low strain partition factor of 0.01% and a low seismogenenic index indicating that aseismic processes dominate. We also analyse the spatio-temporal distribution of hypocentres, using simple models for the evolution of hydraulic diffusivity by (a) isotropic and (b) anisotropic pore-pressure relaxation. The principal axes of the diffusivity or permeability tensor inferred from the spatial distribution of earthquake foci are aligned parallel to the present-day stress field, although the maximum permeability is vertical, whereas the maximum principal stress is horizontal. Our results are consistent with a triggering mechanism that involves (a) seismic shear slip along optimally-oriented pre-existing fractures, (b) a large component of aseismic slip with creep (c) activation of tensile fractures as hydraulic conduits created by both the present-day stress field and by the induced shear slip, both exploiting pre-existing joint sets exposed in borehole data.

A test of the hypothesis that impact-induced fractures are preferred sites for later tectonic activity

Science.gov (United States)

Solomon, Sean C.; Duxbury, Elizabeth D.

1987-01-01

Impact cratering has been an important process in the solar system. The cratering event is generally accompanied by faulting in adjacent terrain. Impact-induced faults are nearly ubiquitous over large areas on the terrestrial planets. The suggestion is made that these fault systems, particularly those associated with the largest impact features are preferred sites for later deformation in response to lithospheric stresses generated by other processes. The evidence is a perceived clustering of orientations of tectonic features either radial or concentric to the crater or basin in question. An opportunity exists to test this suggestion more directly on Earth. The terrestrial continents contain more than 100 known or probable impact craters, with associated geological structures mapped to varying levels of detail. Prime facie evidence for reactivation of crater-induced faults would be the occurrence of earthquakes on these faults in response to the intraplate stress field. Either an alignment of epicenters with mapped fault traces or fault plane solutions indicating slip on a plane approximately coincident with that inferred for a crater-induced fault would be sufficient to demonstrate such an association.

Divergent expression of cytokinin biosynthesis, signaling and catabolism genes underlying differences in feeding sites induced by cyst and root-knot nematodes.

Science.gov (United States)

Dowd, Carola D; Chronis, Demosthenis; Radakovic, Zoran S; Siddique, Shahid; Schmülling, Thomas; Werner, Tomáš; Kakimoto, Tatsuo; Grundler, Florian M W; Mitchum, Melissa G

2017-10-01

Cyst and root-knot nematodes are obligate parasites of economic importance with a remarkable ability to reprogram root cells into unique metabolically active feeding sites. Previous studies have suggested a role for cytokinin in feeding site formation induced by these two types of nematodes, but the mechanistic details have not yet been described. Using Arabidopsis as a host plant species, we conducted a comparative analysis of cytokinin genes in response to the beet cyst nematode (BCN), Heterodera schachtii, and the root-knot nematode (RKN), Meloidogyne incognita. We identified distinct differences in the expression of cytokinin biosynthesis, catabolism and signaling genes in response to infection by BCN and RKN, suggesting differential manipulation of the cytokinin pathway by these two nematode species. Furthermore, we evaluated Arabidopsis histidine kinase receptor mutant lines ahk2/3, ahk2/4 and ahk3/4 in response to RKN infection. Similar to our previous studies with BCN, these lines were significantly less susceptible to RKN without compromising nematode penetration, suggesting a requirement of cytokinin signaling in RKN feeding site formation. Moreover, an analysis of ahk double mutants using CycB1;1:GUS/ahk introgressed lines revealed contrasting differences in the cytokinin receptors mediating cell cycle activation in feeding sites induced by BCN and RKN. © 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd.

One-site versus two-site phacotrabeculectomy: a prospective randomized study

Directory of Open Access Journals (Sweden)

Moschos MM

2015-08-01

Full Text Available Marilita M Moschos,1 Irini P Chatziralli,2 Michael Tsatsos3 1First Department of Ophthalmology, University of Athens, 2Second Department of Ophthalmology, Ophthalmiatrion Athinon, Athens, Greece; 3Department of Ophthalmology, Cambridge University Hospital NHS, Cambridge, UK Purpose: The purpose of this study is to compare the efficacy and safety of one-site and two-site combined phacotrabeculectomy with foldable posterior chamber intraocular lens implantation.Methods: Thirty-four patients (41 eyes with glaucoma and cataract were randomly assigned to undergo either a one-site (22 eyes or a two-site (19 eyes combined procedure. One-site approach consisted of a standard superior phacotrabeculectomy with a limbus-based conjunctival flap, while two-site approach consisted of a clear cornea phacoemulsification and a separate superior trabeculectomy with a limbus-based conjunctival flap.Results: Mean follow-up period was 54 months (standard deviation [SD] 2.3. Mean preoperative intraocular pressure (IOP in the one-site group was 21.3 mmHg (SD 2.8 and in the two-site group was 21.8 mmHg (SD 3.0 (P>0.1. Mean postoperative IOP significantly decreased in both groups compared to the preoperative level and was 15.6 mmHg (SD 3.5 in the one-site group and 14.9 mmHg (SD 2.7 in the two-site group. Three months later, the difference between the two groups was not statistically significant (P=0.058. The one-site group required significantly more medications than the two-site group (P=0.03. Best-corrected visual acuity (BCVA improved similarly in both groups, but there was less postoperative (induced astigmatism in the two-site group in a marginal statistical level (P=0.058. Intra- and postoperative complications were comparable in the two groups.Conclusion: Both techniques yielded similar results concerning final BCVA and IOP reduction. However, the two-site group had less induced astigmatism and a better postoperative IOP control with less required

Multiple sites and actions of gabapentin-induced relief of ongoing experimental neuropathic pain.

Science.gov (United States)

Bannister, Kirsty; Qu, Chaoling; Navratilova, Edita; Oyarzo, Janice; Xie, Jennifer Yanhua; King, Tamara; Dickenson, Anthony H; Porreca, Frank

2017-12-01

Gabapentin (GBP) is a first-line therapy for neuropathic pain, but its mechanisms and sites of action remain uncertain. We investigated GBP-induced modulation of neuropathic pain following spinal nerve ligation (SNL) in rats. Intravenous or intrathecal GBP reversed evoked mechanical hypersensitivity and produced conditioned place preference (CPP) and dopamine (DA) release in the nucleus accumbens (NAc) selectively in SNL rats. Spinal GBP also significantly inhibited dorsal horn wide-dynamic-range neuronal responses to a range of evoked stimuli in SNL rats. By contrast, GBP microinjected bilaterally into the rostral anterior cingulate cortex (rACC), produced CPP, and elicited NAc DA release selectively in SNL rats but did not reverse tactile allodynia and had marginal effects on wide-dynamic-range neuronal activity. Moreover, blockade of endogenous opioid signaling in the rACC prevented intravenous GBP-induced CPP and NAc DA release but failed to block its inhibition of tactile allodynia. Gabapentin, therefore, can potentially act to produce its pain relieving effects by (a) inhibition of injury-induced spinal neuronal excitability, evoked hypersensitivity, and ongoing pain and (b) selective supraspinal modulation of affective qualities of pain, without alteration of reflexive behaviors. Consistent with previous findings of pain relief from nonopioid analgesics, GBP requires engagement of rACC endogenous opioid circuits and downstream activation of mesolimbic reward circuits reflected in learned pain-motivated behaviors. These findings support the partial separation of sensory and affective dimensions of pain in this experimental model and suggest that modulation of affective-motivational qualities of pain may be the preferential mechanism of GBP's analgesic effects in patients.

Database Description - SKIP Stemcell Database | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available List Contact us SKIP Stemcell Database Database Description General information of database Database name SKIP Stemcell Database...rsity Journal Search: Contact address http://www.skip.med.keio.ac.jp/en/contact/ Database classification Human Genes and Diseases Dat...abase classification Stemcell Article Organism Taxonomy Name: Homo sapiens Taxonomy ID: 9606 Database...ks: Original website information Database maintenance site Center for Medical Genetics, School of medicine, ...lable Web services Not available URL of Web services - Need for user registration Not available About This Database Database

Mechanism of cluster DNA damage repair in response to high-atomic number and energy particles radiation

Energy Technology Data Exchange (ETDEWEB)

Asaithamby, Aroumougame, E-mail: Aroumougame.Asaithamy@UTsouthwestern.edu [Division of Molecular Radiation Biology, Department of Radiation Oncology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390 (United States); Chen, David J., E-mail: David.Chen@UTsouthwestern.edu [Division of Molecular Radiation Biology, Department of Radiation Oncology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390 (United States)

2011-06-03

Low-linear energy transfer (LET) radiation (i.e., {gamma}- and X-rays) induces DNA double-strand breaks (DSBs) that are rapidly repaired (rejoined). In contrast, DNA damage induced by the dense ionizing track of high-atomic number and energy (HZE) particles is slowly repaired or is irreparable. These unrepaired and/or misrepaired DNA lesions may contribute to the observed higher relative biological effectiveness for cell killing, chromosomal aberrations, mutagenesis, and carcinogenesis in HZE particle irradiated cells compared to those treated with low-LET radiation. The types of DNA lesions induced by HZE particles have been characterized in vitro and usually consist of two or more closely spaced strand breaks, abasic sites, or oxidized bases on opposing strands. It is unclear why these lesions are difficult to repair. In this review, we highlight the potential of a new technology allowing direct visualization of different types of DNA lesions in human cells and document the emerging significance of live-cell imaging for elucidation of the spatio-temporal characterization of complex DNA damage. We focus on the recent insights into the molecular pathways that participate in the repair of HZE particle-induced DSBs. We also discuss recent advances in our understanding of how different end-processing nucleases aid in repair of DSBs with complicated ends generated by HZE particles. Understanding the mechanism underlying the repair of DNA damage induced by HZE particles will have important implications for estimating the risks to human health associated with HZE particle exposure.

Mechanism of cluster DNA damage repair in response to high-atomic number and energy particles radiation

International Nuclear Information System (INIS)

Asaithamby, Aroumougame; Chen, David J.

2011-01-01

Low-linear energy transfer (LET) radiation (i.e., γ- and X-rays) induces DNA double-strand breaks (DSBs) that are rapidly repaired (rejoined). In contrast, DNA damage induced by the dense ionizing track of high-atomic number and energy (HZE) particles is slowly repaired or is irreparable. These unrepaired and/or misrepaired DNA lesions may contribute to the observed higher relative biological effectiveness for cell killing, chromosomal aberrations, mutagenesis, and carcinogenesis in HZE particle irradiated cells compared to those treated with low-LET radiation. The types of DNA lesions induced by HZE particles have been characterized in vitro and usually consist of two or more closely spaced strand breaks, abasic sites, or oxidized bases on opposing strands. It is unclear why these lesions are difficult to repair. In this review, we highlight the potential of a new technology allowing direct visualization of different types of DNA lesions in human cells and document the emerging significance of live-cell imaging for elucidation of the spatio-temporal characterization of complex DNA damage. We focus on the recent insights into the molecular pathways that participate in the repair of HZE particle-induced DSBs. We also discuss recent advances in our understanding of how different end-processing nucleases aid in repair of DSBs with complicated ends generated by HZE particles. Understanding the mechanism underlying the repair of DNA damage induced by HZE particles will have important implications for estimating the risks to human health associated with HZE particle exposure.

Tiron Inhibits UVB-Induced AP-1 Binding Sites Transcriptional Activation on MMP-1 and MMP-3 Promoters by MAPK Signaling Pathway in Human Dermal Fibroblasts.

Directory of Open Access Journals (Sweden)

Jing Lu

Full Text Available Recent research found that Tiron was an effective antioxidant that could act as the intracellular reactive oxygen species (ROS scavenger or alleviate the acute toxic metal overload in vivo. In this study, we investigated the inhibitory effect of Tiron on matrix metalloproteinase (MMP-1 and MMP-3 expression in human dermal fibroblast cells. Western blot and ELISA analysis revealed that Tiron inhibited ultraviolet B (UVB-induced protein expression of MMP-1 and MMP-3. Real-time quantitative PCR confirmed that Tiron could inhibit UVB-induced mRNA expression of MMP-1 and MMP-3. Furthermore, Tiron significantly blocked UVB-induced activation of the MAPK signaling pathway and activator protein (AP-1 in the downstream of this transduction pathway in fibroblasts. Through the AP-1 binding site mutation, it was found that Tiron could inhibit AP-1-induced upregulation of MMP-1 and MMP-3 expression through blocking AP-1 binding to the AP-1 binding sites in the MMP-1 and MMP-3 promoter region. In conclusion, Tiron may be a novel antioxidant for preventing and treating skin photoaging UV-induced.

Spectroscopic study of site selective DNA damage induced by intense soft X-rays

CERN Document Server

Fujii, K

2003-01-01

To investigate the mechanisms of DNA damage induced by direct photon impact, we observed the near edge X-ray absorption fine structures (NEXAFS) of DNA nucleobases using monochromatic synchrotron soft X-rays around nitrogen and oxygen K-shell excitation regions. Each spectrum obtained has unique structure corresponding to pi* excitation of oxygen or nitrogen 1s electron. These aspects open a way of nucleobase-selective photo-excitation in a DNA molecule using high resolution monochromatized soft X-rays. From the analysis of polarization-dependent intensities of the pi* resonance peak, it is clarified that adenine, guanine an uracil form orientated surface structure. Furthermore from the direct measurement of positive ions desorbed from photon irradiated DNA components, it is revealed that the sugar moiety is a fragile site in a DNA molecule. (author)

Temperature and electrolyte optimization of the α-hemolysin latch sensing zone for detection of base modification in double-stranded DNA.

Science.gov (United States)

Johnson, Robert P; Fleming, Aaron M; Jin, Qian; Burrows, Cynthia J; White, Henry S

2014-08-19

The latch region of the wild-type protein pore α-hemolysin (α-HL) constitutes a sensing zone for individual abasic sites (and furan analogs) in double-stranded DNA (dsDNA). The presence of an abasic site or furan within a DNA duplex, electrophoretically captured in the α-HL vestibule and positioned at the latch region, can be detected based on the current blockage prior to duplex unzipping. We investigated variations in blockage current as a function of temperature (12-35°C) and KCl concentration (0.15-1.0 M) to understand the origin of the current signature and to optimize conditions for identifying the base modification. In 1 M KCl solution, substitution of a furan for a cytosine base in the latch region results in an ∼ 8 kJ mol(-1) decrease in the activation energy for ion transport through the protein pore. This corresponds to a readily measured ∼ 2 pA increase in current at room temperature. Optimal resolution for detecting the presence of a furan in the latch region is achieved at lower KCl concentrations, where the noise in the measured blockage current is significantly lower. The noise associated with the blockage current also depends on the stability of the duplex (as measured from the melting temperature), where a greater noise in the measured blockage current is observed for less stable duplexes. Copyright © 2014 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Drug-induced sleep endoscopy in the identification of obstruction sites in patients with obstructive sleep apnea: a systematic review.

Science.gov (United States)

Viana, Alonço da Cunha; Thuler, Luiz Claudio Santos; Araújo-Melo, Maria Helena de

2015-01-01

Obstructive sleep apnea syndrome has multifactorial causes. Although indications for surgery are evaluated by well-known diagnostic tests in the awake state, these do not always correlate with satisfactory surgical results. To undertake a systematic review on endoscopy during sleep, as one element of the diagnosis routine, aiming to identify upper airway obstruction sites in adult patients with OSAS. By means of electronic databases, a systematic review was performed of studies using drug-induced sleep endoscopy to identify obstruction sites in patients with OSAS. Ten articles were selected that demonstrated the importance of identifying multilevel obstruction, especially in relation to retrolingual and laryngeal collapse in OSAS. DISE is an additional method to reveal obstruction sites that have not been detected in awake patients. Copyright © 2015 Associação Brasileira de Otorrinolaringologia e Cirurgia Cérvico-Facial. Published by Elsevier Editora Ltda. All rights reserved.

Downregulation of hPMC2 imparts chemotherapeutic sensitivity to alkylating agents in breast cancer cells.

Science.gov (United States)

Krishnamurthy, Nirmala; Liu, Lili; Xiong, Xiahui; Zhang, Junran; Montano, Monica M

2015-01-01

Triple negative breast cancer cell lines have been reported to be resistant to the cyotoxic effects of temozolomide (TMZ). We have shown previously that a novel protein, human homolog of Xenopus gene which Prevents Mitotic Catastrophe (hPMC2) has a role in the repair of estrogen-induced abasic sites. Our present study provides evidence that downregulation of hPMC2 in MDA-MB-231 and MDA-MB-468 breast cancer cells treated with temozolomide (TMZ) decreases cell survival. This increased sensitivity to TMZ is associated with an increase in number of apurinic/apyrimidinic (AP) sites in the DNA. We also show that treatment with another alkylating agent, BCNU, results in an increase in AP sites and decrease in cell survival. Quantification of western blot analyses and immunofluorescence experiments reveal that treatment of hPMC2 downregulated cells with TMZ results in an increase in γ-H2AX levels, suggesting an increase in double strand DNA breaks. The enhancement of DNA double strand breaks in TMZ treated cells upon downregulation of hPCM2 is also revealed by the comet assay. Overall, we provide evidence that downregulation of hPMC2 in breast cancer cells increases cytotoxicity of alkylating agents, representing a novel mechanism of treatment for breast cancer. Our data thus has important clinical implications in the management of breast cancer and brings forth potentially new therapeutic strategies.

Shape-selective recognition of DNA abasic sites by metallohelices: inhibition of human AP endonuclease 1

Czech Academy of Sciences Publication Activity Database

Malina, Jaroslav; Scott, P.; Brabec, Viktor

2015-01-01

Roč. 43, č. 11 (2015), s. 5297-5306 ISSN 1362-4962 R&D Projects: GA ČR(CZ) GAP205/11/0856 Institutional support: RVO:68081707 Keywords : BASE EXCISION-REPAIR * CYTOSINE OPPOSITE * BINDING Subject RIV: BO - Biophysics

Ground Combat Training Squadron Complex Final Environmental Assessment

Science.gov (United States)

2011-08-01

Squadron E nvironm ental A ssessm ent Page 1-6 Eglin A ir Force Base, FL Final Figure 1-2. R esources N ot C arried Forw ard for D etailed A...Base, FL Final Figure 3-1. W ater R esources w ithin or near the Proposed A ction or A lternative L ocations Legend c::J Project Sites 1 00...bat T raining Squadron E nvironm ental A ssessm ent Page 3-14 Eglin A ir Force Base, FL Final Figure 3-3. B iological R esources w ithin or

DNA translocation by human uracil DNA glycosylase: the case of single-stranded DNA and clustered uracils.

Science.gov (United States)

Schonhoft, Joseph D; Stivers, James T

2013-04-16

Human uracil DNA glycosylase (hUNG) plays a central role in DNA repair and programmed mutagenesis of Ig genes, requiring it to act on sparsely or densely spaced uracil bases located in a variety of contexts, including U/A and U/G base pairs, and potentially uracils within single-stranded DNA (ssDNA). An interesting question is whether the facilitated search mode of hUNG, which includes both DNA sliding and hopping, changes in these different contexts. Here we find that hUNG uses an enhanced local search mode when it acts on uracils in ssDNA, and also, in a context where uracils are densely clustered in duplex DNA. In the context of ssDNA, hUNG performs an enhanced local search by sliding with a mean sliding length larger than that of double-stranded DNA (dsDNA). In the context of duplex DNA, insertion of high-affinity abasic product sites between two uracil lesions serves to significantly extend the apparent sliding length on dsDNA from 4 to 20 bp and, in some cases, leads to directionally biased 3' → 5' sliding. The presence of intervening abasic product sites mimics the situation where hUNG acts iteratively on densely spaced uracils. The findings suggest that intervening product sites serve to increase the amount of time the enzyme remains associated with DNA as compared to nonspecific DNA, which in turn increases the likelihood of sliding as opposed to falling off the DNA. These findings illustrate how the search mechanism of hUNG is not predetermined but, instead, depends on the context in which the uracils are located.

Background estimation of cosmic-ray induced neutrons in Chooz site water veto tank for possible future Ricochet Deployment

Science.gov (United States)

Silva, James

2017-09-01

The Ricochet experiment seeks to measure Coherent (neutral-current) Elastic Neutrino-Nucleus Scattering (CE νNS) using metallic superconducting and germanium semi-conducting detectors with sub-keV thresholds placed near a neutrino source such as the Chooz Nuclear Reactor Complex. In this poster, we present an estimate of the flux of cosmic-ray induced neutrons, which represent an important background in any (CE νNS) search, based on reconstructed cosmic ray data from the Chooz Site. We have simulated a possible Ricochet deployment at the Chooz site in GEANT4 focusing on the spallation neutrons generated when cosmic rays interact with the water tank veto that would surround our detector. We further simulate and discuss the effectiveness of various shielding configurations for optimizing the background levels for a future Ricochet deployment.

Tracking Human-Induced Landscape Disturbance at the Nasca Lines UNESCO World Heritage Site in Peru with COSMO-SkyMed InSAR

Directory of Open Access Journals (Sweden)

Francesca Cigna

2018-04-01

Full Text Available The “Lines and Geoglyphs of Nasca and Palpa” in Peru are among the most well-known UNESCO World Heritage Sites globally, and an exemplar of site where heritage assets cannot be separated from their natural and anthropogenic environment. The site is exposed to interactions with natural processes, as well as human presence. In this work, 3-m resolution synthetic aperture radar (SAR StripMap HIMAGE HH-polarised scenes acquired by the X-band COSMO-SkyMed constellation are exploited to track two events of human-induced landscape disturbance that occurred in December 2014 and January 2018. Pre-, cross-, and post-event interferometric SAR (InSAR pairs characterised by small temporal and normal baselines allow the detection of temporal decorrelation associated with the two events, the extent and time reference of which match with online photographic and video evidence, published literature, web news, and press releases by the Ministry of Culture in Peru. Further elements enhancing the understanding of the 2018 event come from 10-m resolution Sentinel-2B satellite data that reveal the occurrence of apparent changes of surface reflectance due to uncovering of the light grey-yellow clay underneath the darker pebble constituting the fragile surface of the Pampa de Jumana. This scientific study confirms that SAR imagery archives, such as those being built by COSMO-SkyMed for Nasca, prove valuable for the retrospective analysis and digital recording of human-induced landscape disturbance events from space. These archives therefore act as essential sources of geospatial information on the conservation history of heritage sites and assets.

ANTHROPOGENIC POLLEN INDICATORS (API FROM ARCHAEOLOGICAL SITES AS LOCAL EVIDENCE OF HUMAN-INDUCED ENVIRONMENTS IN THE ITALIAN PENINSULA

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A. M. Mercuri

2013-04-01

Full Text Available Pollen data from twenty-six archaeological sites are reviewed to investigate the development of human-induced environments through the presence of selected Anthropogenic Pollen Indicators (API. The sites are located in six Italian regions - Veneto, Emilia Romagna, Tuscany, Basilicata, Calabria, and Sicily - and in the Republic of San Marino. Their chronology spans from the Bronze to the Renaissance ages, from approximately 4200 to 500 years BP. The API which are common in these sites are properly considered important markers of human activity and anthropization in the Mediterranean area. The most frequent API taxa in pollen spectra are seven: Artemisia, Centaurea, Cichorieae and Plantago are ubiquitous and therefore they have the major relevance, followed by cereals and Urtica, and by Trifolium type. The spread of plants producing these pollen grains is sometimes marked by high percentage values in pollen spectra. Pollen records show that, as expected, cereals and wild synanthropic herbs were widespread near archaeological sites but local differences are evident. Ecological and chrono-cultural reasons may be at the base of the observed differences. In general, the synanthropic plants well represent the xeric environments that developed as a result of the continuous human pressure and changes in soil compositions. These changes have occurred especially during the mid and late Holocene.

Retina, Retinol, Retinal and the Natural History of Vitamin A as a Light Sensor

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Hui Sun

2012-12-01

Full Text Available Light is both the ultimate energy source for most organisms and a rich information source. Vitamin A-based chromophore was initially used in harvesting light energy, but has become the most widely used light sensor throughout evolution from unicellular to multicellular organisms. Vitamin A-based photoreceptor proteins are called opsins and have been used for billions of years for sensing light for vision or the equivalent of vision. All vitamin A-based light sensors for vision in the animal kingdom are G-protein coupled receptors, while those in unicellular organisms are light-gated channels. This first major switch in evolution was followed by two other major changes: the switch from bistable to monostable pigments for vision and the expansion of vitamin A’s biological functions. Vitamin A’s new functions such as regulating cell growth and differentiation from embryogenesis to adult are associated with increased toxicity with its random diffusion. In contrast to bistable pigments which can be regenerated by light, monostable pigments depend on complex enzymatic cycles for regeneration after every photoisomerization event. Here we discuss vitamin A functions and transport in the context of the natural history of vitamin A-based light sensors and propose that the expanding functions of vitamin A and the choice of monostable pigments are the likely evolutionary driving forces for precise, efficient, and sustained vitamin A transport.

Pre-steady-state fluorescence analysis of damaged DNA transfer from human DNA glycosylases to AP endonuclease APE1.

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Kuznetsova, Alexandra A; Kuznetsov, Nikita A; Ishchenko, Alexander A; Saparbaev, Murat K; Fedorova, Olga S

2014-10-01

DNA glycosylases remove the modified, damaged or mismatched bases from the DNA by hydrolyzing the N-glycosidic bonds. Some enzymes can further catalyze the incision of a resulting abasic (apurinic/apyrimidinic, AP) site through β- or β,δ-elimination mechanisms. In most cases, the incision reaction of the AP-site is catalyzed by special enzymes called AP-endonucleases. Here, we report the kinetic analysis of the mechanisms of modified DNA transfer from some DNA glycosylases to the AP endonuclease, APE1. The modified DNA contained the tetrahydrofurane residue (F), the analogue of the AP-site. DNA glycosylases AAG, OGG1, NEIL1, MBD4(cat) and UNG from different structural superfamilies were used. We found that all DNA glycosylases may utilise direct protein-protein interactions in the transient ternary complex for the transfer of the AP-containing DNA strand to APE1. We hypothesize a fast "flip-flop" exchange mechanism of damaged and undamaged DNA strands within this complex for monofunctional DNA glycosylases like MBD4(cat), AAG and UNG. Bifunctional DNA glycosylase NEIL1 creates tightly specific complex with DNA containing F-site thereby efficiently competing with APE1. Whereas APE1 fast displaces other bifunctional DNA glycosylase OGG1 on F-site thereby induces its shifts to undamaged DNA regions. Kinetic analysis of the transfer of DNA between human DNA glycosylases and APE1 allows us to elucidate the critical step in the base excision repair pathway. Copyright © 2014 Elsevier B.V. All rights reserved.

cGAS-Mediated Innate Immunity Spreads Intercellularly through HIV-1 Env-Induced Membrane Fusion Sites.

Science.gov (United States)

Xu, Shuting; Ducroux, Aurélie; Ponnurangam, Aparna; Vieyres, Gabrielle; Franz, Sergej; Müsken, Mathias; Zillinger, Thomas; Malassa, Angelina; Ewald, Ellen; Hornung, Veit; Barchet, Winfried; Häussler, Susanne; Pietschmann, Thomas; Goffinet, Christine

2016-10-12

Upon sensing cytoplasmic retroviral DNA in infected cells, cyclic GMP-AMP (cGAMP) synthase (cGAS) produces the cyclic dinucleotide cGAMP, which activates STING to trigger a type I interferon (IFN) response. We find that membrane fusion-inducing contact between donor cells expressing the HIV envelope (Env) and primary macrophages endogenously expressing the HIV receptor CD4 and coreceptor enable intercellular transfer of cGAMP. This cGAMP exchange results in STING-dependent antiviral IFN responses in target macrophages and protection from HIV infection. Furthermore, under conditions allowing cell-to-cell transmission of HIV-1, infected primary T cells, but not cell-free virions, deliver cGAMP to autologous macrophages through HIV-1 Env and CD4/coreceptor-mediated membrane fusion sites and induce a STING-dependent, but cGAS-independent, IFN response in target cells. Collectively, these findings identify an infection-specific mode of horizontal transfer of cGAMP between primary immune cells that may boost antiviral responses, particularly in infected tissues in which cell-to-cell transmission of virions exceeds cell-free infection. Copyright © 2016 Elsevier Inc. All rights reserved.

Histone peptide AKRHRK enhances H2O2-induced DNA damage and alters its site specificity

International Nuclear Information System (INIS)

Midorikawa, Kaoru; Murata, Mariko; Kawanishi, Shosuke

2005-01-01

Histone proteins are involved in compaction of DNA and the protection of cells from oxygen toxicity. However, several studies have demonstrated that the metal-binding histone reacts with H 2 O 2 , leading to oxidative damage to a nucleobase. We investigated whether histone can accelerate oxidative DNA damage, using a minimal model for the N-terminal tail of histone H4, CH 3 CO-AKRHRK-CONH 2 , which has a metal-binding site. This histone peptide enhanced DNA damage induced by H 2 O 2 and Cu(II), especially at cytosine residues, and induced additional DNA cleavage at the 5'-guanine of GGG sequences. The peptide also enhanced the formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine and ESR spin-trapping signal from H 2 O 2 and Cu(II). Cyclic redox reactions involving histone-bound Cu(II) and H 2 O 2 , may give rise to multiple production of radicals leading to multiple hits in DNA. It is noteworthy that the histone H4 peptide with specific sequence AKRHRK can cause DNA damage rather than protection under metal-overloaded condition

Selection and investigation of sites for the disposal of radioactive wastes in hydraulically induced subsurface fractures

International Nuclear Information System (INIS)

Sun, R.J.

1982-01-01

Injection of intermediate-level radioactive wastes (specific activity of less than 6 x 10 3 μCi/mL, consisting mainly of radionuclides, such as strontium and cesium, having half-lives of less than 50 years) mixed with cement into a thick shale formation is a promising and feasible disposal method. Hydraulic fracturing provides openings in the shale to accommodate the wastes. Ion exchange and radionuclide-adsorption materials can be added to the grout during mixing to further increase the radionuclide-retaining capacity of the grout. After solidification of the grout, the injected wastes become an integral part of the shale formation, and therefore the wastes will remain at depth and in place as long as the injection zone is not subjected to erosion and dissolution. Problems concerning safety of the disposal method are (1) the potential for inducing vertical fractures, (2) phase separation during and after the injections, (3) the reliability of methods for determining the orientation of induced fractures, (4) the possibility of triggering earthquakes, and (5) radionuclides being leached and transported by ground water. Theoretical considerations about inducing nearly horizontal bedding-plane fractures in shale are discussed, as are field procedures for site selection, safety, and the monitoring and operation of radioactive waste disposal. Case histories are used as examples to demonstrate the application of the theory and techniques of field operations

New Reactor Siting in Finland, Hanhikivi Site in Pyhaejoki - STUK preliminary safety assessment

International Nuclear Information System (INIS)

Nevalainen, Janne

2013-01-01

STUK has performed a preliminary assessment of the Decision-in-Principle on the Fennovoima application. A variety of factors must be considered in the selection of a site, including effects of the site on the plant design and the effects of the plant on the site environment. These include external hazards, both natural and human-induced. Since this is a new site, an extensive siting process is followed, that can include an EIA. A site survey is performed to identify candidate sites, after investigating a large region and rejecting unsuitable sites. The remaining sites are then screened and compared on the basis of safety and other considerations to select one or more preferred sites. Natural hazards include geology, seismology, hydrology and meteorology. Offshore ice will be a particular hazard for this plant, since the site is on average only 1.5 m above sea level. The design basis earthquake corresponds to a return frequency of 100,000 years, with 50 % confidence. The existing sites in southern Finland used a design peak ground acceleration of 0.1 g with the ground response spectrum maximum at 10 Hz. The candidate sites in northern Finland will require a peak ground acceleration of 0.2 g with the ground response spectrum maximum at 25 Hz

Subsurface imaging of water electrical conductivity, hydraulic permeability and lithology at contaminated sites by induced polarization

Science.gov (United States)

Maurya, P. K.; Balbarini, N.; Møller, I.; Rønde, V.; Christiansen, A. V.; Bjerg, P. L.; Auken, E.; Fiandaca, G.

2018-05-01

At contaminated sites, knowledge about geology and hydraulic properties of the subsurface and extent of the contamination is needed for assessing the risk and for designing potential site remediation. In this study, we have developed a new approach for characterizing contaminated sites through time-domain spectral induced polarization. The new approach is based on: (1) spectral inversion of the induced polarization data through a reparametrization of the Cole-Cole model, which disentangles the electrolytic bulk conductivity from the surface conductivity for delineating the contamination plume; (2) estimation of hydraulic permeability directly from the inverted parameters using a laboratory-derived empirical equation without any calibration; (3) the use of the geophysical imaging results for supporting the geological modelling and planning of drilling campaigns. The new approach was tested on a data set from the Grindsted stream (Denmark), where contaminated groundwater from a factory site discharges to the stream. Two overlapping areas were covered with seven parallel 2-D profiles each, one large area of 410 m × 90 m (5 m electrode spacing) and one detailed area of 126 m × 42 m (2 m electrode spacing). The geophysical results were complemented and validated by an extensive set of hydrologic and geologic information, including 94 estimates of hydraulic permeability obtained from slug tests and grain size analyses, 89 measurements of water electrical conductivity in groundwater, and four geological logs. On average the IP-derived and measured permeability values agreed within one order of magnitude, except for those close to boundaries between lithological layers (e.g. between sand and clay), where mismatches occurred due to the lack of vertical resolution in the geophysical imaging. An average formation factor was estimated from the correlation between the imaged bulk conductivity values and the water conductivity values measured in groundwater, in order to

Increased expression of aphidicolin-induced common fragile sites in Tourette syndrome: The key to understand the genetics of comorbid phenotypes?

Energy Technology Data Exchange (ETDEWEB)

Gericke, G.S.; Simonic, I.; Cloete, E.; Becker, P.J. [Univ. of Pretoria (South Africa)

1996-02-16

In a comparison of 80 common aphidicolin-induced fragile sites (FS) between 26 DSM-IV Tourette syndrome (TS) and 24 control individuals, the mean of the summed break frequencies following mild aphidicolin pretreatment was significantly higher in TS individuals than in controls (P < 0.001). Other breakpoints encountered during this study, i.e., random breaks, breaks corresponding to rare FS, and breakpoints recorded by others but not listed as common FS according to the Chromosome Coordinating Meeting were listed as category II breakpoints. By using the most significantly different mean FS breakage figures between TS and control individuals, further stepwise discriminant analysis allowed identification of TS individuals from only a few sites in both the common FS and category II breakpoint groups. Future research needs to focus on confirmation of altered common fragile site expression in association with behavioral variation, whether expression of certain discriminatory sites concurs with specific comorbid disorder expression; the nature of the molecular alterations at these FS and the implications of a genomic instability phenotype for the mapping of a primary TS gene or genes. 45 refs., 1 fig., 2 tabs.

Exterior Site Occupancy Infers Chloride-Induced Proton Gating in a Prokaryotic Homolog of the ClC Chloride Channel

Science.gov (United States)

Bostick, David L.; Berkowitz, Max L.

2004-01-01

The ClC family of anion channels mediates the efficient, selective permeation of Cl− across the biological membranes of living cells under the driving force of an electrochemical gradient. In some eukaryotes, these channels are known to exhibit a unique gating mechanism, which appears to be triggered by the permeant Cl− anion. We infer details of this gating mechanism by studying the free energetics of Cl− occupancy in the pore of a prokaryotic ClC homolog. These free energetics were gleaned from 30 ns of molecular dynamics simulation on an ∼133,000-atom system consisting of a hydrated membrane embedded StClC transporter. The binding sites for Cl− in the transporter were determined for the cases where the putative gating residue, Glu148, was protonated and unprotonated. When the glutamate gate is protonated, Cl− favorably occupies an exterior site, Sext, to form a queue of anions in the pore. However, when the glutamate gate is unprotonated, Cl− cannot occupy this site nor, consequently, pass through the pore. An additional, previously undetected, site was found in the pore near the outer membrane that exists regardless of the protonation state of Glu148. Although this suggests that, for the prokaryotic homolog, protonation of Glu148 may be the first step in transporting Cl− at the expense of H+ transport in the opposite direction, an evolutionary argument might suggest that Cl− opens the ClC gate in eukaryotic channels by inducing the conserved glutamate's protonation. During an additional 20 ns free dynamics simulation, the newly discovered outermost site, Sout, and the innermost site, Sint, were seen to allow spontaneous exchange of Cl− ions with the bulk electrolyte while under depolarization conditions. PMID:15345547

Evolution of Metal(Loid) Binding Sites in Transcriptional Regulators

Energy Technology Data Exchange (ETDEWEB)

Ordonez, E.; Thiyagarajan, S.; Cook, J.D.; Stemmler, T.L.; Gil, J.A.; Mateos, L.M.; Rosen, B.P.

2009-05-22

Expression of the genes for resistance to heavy metals and metalloids is transcriptionally regulated by the toxic ions themselves. Members of the ArsR/SmtB family of small metalloregulatory proteins respond to transition metals, heavy metals, and metalloids, including As(III), Sb(III), Cd(II), Pb(II), Zn(II), Co(II), and Ni(II). These homodimeric repressors bind to DNA in the absence of inducing metal(loid) ion and dissociate from the DNA when inducer is bound. The regulatory sites are often three- or four-coordinate metal binding sites composed of cysteine thiolates. Surprisingly, in two different As(III)-responsive regulators, the metalloid binding sites were in different locations in the repressor, and the Cd(II) binding sites were in two different locations in two Cd(II)-responsive regulators. We hypothesize that ArsR/SmtB repressors have a common backbone structure, that of a winged helix DNA-binding protein, but have considerable plasticity in the location of inducer binding sites. Here we show that an As(III)-responsive member of the family, CgArsR1 from Corynebacterium glutamicum, binds As(III) to a cysteine triad composed of Cys{sup 15}, Cys{sup 16}, and Cys{sup 55}. This binding site is clearly unrelated to the binding sites of other characterized ArsR/SmtB family members. This is consistent with our hypothesis that metal(loid) binding sites in DNA binding proteins evolve convergently in response to persistent environmental pressures.

Inacessible Andean sites reveal land-use induced stabilisation of soil organic carbon

Science.gov (United States)

Heitkamp, Felix; Maqsood, Shafique; Sylvester, Steven; Kessler, Michael; Jungkunst, Hermann

2015-04-01

Human activity affects properties and development of ecosystems across the globe to such a degree that it is challenging to get baseline values for undisturbed ecosystems. This is especially true for soils, which are affected by land-use history and hold a legacy of past human interventions. Therefore, it is still largely unknown how soil would have developed "naturally" and if processes of organic matter stabilisation would be different in comparison to managed soils. Here, we show undisturbed soil development, i.e., the processes of weathering and accumulation of soil organic carbon (SOC), by comparing pristine with grazed sites in the high Andes (4500 m) of southern Peru. We located study plots on a large ledge (0.2 km²) that is only accessible with mountaineering equipment. Plots with pristine vegetation were compared to rangeland plots that were constantly under grazing management for at least four millennia. All "state factors"; climate, potential biota, topography, parent material and time; besides "land-use" were, therefore, identical. Vegetation change, induced by grazing management, led to lower vegetation cover of the soil, thereby increasing soil surface temperatures and soil acidification. Both factors increased weathering in rangeland soils, as indicated by the presence of pedogenic oxides, especially amorphous Al-(oxy)hydroxides (oxalate-extractable Al). Higher losses of base cations (K, Na, Ca) and lower pH-values were related to a low base saturation of exchange sites in rangelands. Therefore, rangeland soils were classified as Umbrisol, whereas soils under pristine vegetation were classified as Phaeozeme. All profiles were rich in SOC (100 to 126 g kg-1) with no significant differences in concentrations or stocks. SOC of rangeland soils was, however, less available for microorganisms (proportion of microbial C on SOC: 1.8 vs. 0.6% in pristine and rangeland soils, respectively) and showed higher stability against thermal degradation. Reasons for

Thermally induced A'-A site exchange in novel layered perovskites Ag2[Ca1.5M3O10] (M = Nb, Ta).

Science.gov (United States)

Bhuvanesh, Nattamai S P; Woodward, Patrick M

2002-12-04

We have synthesized and characterized new layered perovskites Ag2[A1.5M3O10] (A = Ca, M = Nb, Ta), from their lithium analogues, by soft-chemical ion exchange. These oxides show topotactic irreversible thermally induced A'-A site exchange, resulting in Ag1.1Ca0.9[Ca0.6Ag0.9M3O10], conferred from our high-temperature X-ray and ionic conductivity studies. The latter phases are the first compounds where Ag+ ions reside in both A' and A sites in layered perovskites. The absence of similar phase transition for A = Sr suggests that these transitions strongly depend on the size, charge, and the coordination preference of A' and A cations. This result provides a new synthetic tool for modifying the occupation of the 12-coordinate A site of layered perovskites using soft chemical routes.

Implications for the formation of abasic sites following modification of polydeoxycytidylic acid by acrolein in vitro

International Nuclear Information System (INIS)

Smith, R.A.; Sysel, I.A.; Tibbels, T.S.; Cohen, S.M.

1988-01-01

Polydeoxycytidylic acid (poly dC) was incubated with excess acrolein. A Nensorb 20 nucleic acid purification cartridge was used to bind the polymeric material in the poly dC/acrolein reaction mixture. The non-polymeric material eluted from this column had a UV absorbance four times higher than that of the control. The flourescence spectrum of the eluted material did not correspond to that of unmodified cytosine. Separate aliquots of the reaction mixture were digested to deoxynucleotide 3 ' -monophosphates by incubation with micrococcal nuclease and spleen phosphodiesterase. The products were converted to 3 2P-labelled deoxynucleotide 3 ' ,5-biphosphates by incubation with T4 polynucleotide kinase and excess [γ- 3 2P]ATP. The ' -monophosphate was selectively removed by incubation with nuclease P1. Two dimensional thin-layer chromatography (TLC) on polyethyleneimine cellulose (PEI)-cellulose and detection of 3 2P-labeled deoxynucleotide 5 ' -monophosphates by autoradiography failed to provide evidence for the formation of an acrolein adduct of deoxycytidine 5'-monophosphate. When acrolein-modified deoxycytidine 5 ' -monophosphate, was detected. These data show that acrolein-modified deoxycytidine 3 ' -monophosphates are substrates for 3 2P labeling by T4 polynucleotide kinase and are stable under the assay conditions employed

Detection of endonuclease III- and 8-oxoguanine glycosylase-sensitive base modifications in γ-irradiated DNA and cells by the aldehyde reactive probe (ARP) assay

International Nuclear Information System (INIS)

Mohsin Ali, M.; Kurisu, Satofumi; Yoshioka, Yoshihiro; Terato, Hiroaki; Ohyama, Yoshihiko; Ide Hiroshi; Kubo, Kihei

2004-01-01

Ionizing radiation generates diverse DNA lesions that differentially induce cell death and mutations. In the present study, calf thymus DNA (400 μg/ml) and HeLa cells were irradiated by 60 Co γ-rays, and abasic (AP) sites and endonuclease (Endo) III- and 8-oxoguanine glycosylase (hOGG1)-sensitive base modifications in DNA were quantitated by the aldehyde reactive probe (ARP) assay. The irradiation of calf thymus DNA in phosphate buffer generated 91 Endo III- and 100 hOGG1-sensitive base modifications and 110 AP sites per 10 6 base pairs (bp) per Gy. The yield of the lesions in Tris buffer was 41- to 91-fold lower than that in phosphate, demonstrating a radioprotective effect of Tris. The HeLa cell chromosomal DNA contained 12 Endo III- and 3.8 hOGG1-sensitive base modifications and less than 1 AP sites per 10 6 bp as endogenous damage, and their level was increased by irradiation. The yields of the damage at 1 Gy (roughly equivalent to the lethal dose of HeLa cells [1.6-1.8 Gy]) were 0.13 Endo III, 0.091 hOGG1, and 0.065 AP sites per 10 6 bp, showing that irradiation with a lethal dose brought about only a marginal increase in base damage relative to an endogenous one. A comparison of the present data with those reported for DNA strand breaks supports the primary importance of double-strand breaks and clustered lesions as lethal damages formed by ionizing radiation. (author)

Tracking the Chemical Transformations at the Brønsted Acid Site upon Water-Induced Deprotonation in a Zeolite Pore

Energy Technology Data Exchange (ETDEWEB)

Vjunov, Aleksei; Wang, Meng; Govind, Niranjan; Huthwelker, Thomas; Shi, Hui; Mei, Donghai; Fulton, John L.; Lercher, Johannes A.

2017-10-23

We report the structural changes induced by Brønsted acidic site deprotonation in a zeolite with MFI structure as a function of temperature up to 430°C using in situ Al K-edge X-ray absorption fine structure spectroscopy (XAFS). At ambient conditions, the protons are present as hydrated hydronium ions (H3O+(H2O)n) that are ion-paired to the anionic, Al tetrahedral (T) site. At elevated temperatures, loss of water molecules hydrating the hydronium ions leads to an unstable free hydronium ion that disso-ciates to form the hydroxylated T-site. The formation of this (-O3)-Al-(OH-) species leads to the elongation of one of the four Al-O bonds and causes significant distortion of the tetrahedral symmetry about the Al atom. This distortion leads to the appearance of new pre-edge features in the Al K-edge X-ray absorption near edge structure (XANES) spectra. The pre-edge peak assignment is confirmed by time-dependent density functional theory calculation of the XANES spectrum. The XANES spectra are also sensitive to solutes or solvent that are in proximity to the T-site. A second structural transition occurs at about the same temperature, namely the conversion of a minor fraction of extra-framework octahedral Al present in the sample at ambient conditions to a tetrahedral species through the de-coordination of H2O-ligands. Both IR spectroscopy and thermogravimetric analysis (TGA) are further used to confirm the overall chemical transformation of the T-site.

Geo-Proxy-Based Site Classification for Regional Zonation of Seismic Site Effects in South Korea

Directory of Open Access Journals (Sweden)

Chang-Guk Sun

2018-02-01

Full Text Available Seismic site effects and topographic effects related to ground motion occur during an earthquake due to site-specific geotechnical or geological characteristics, including the geological or geographical structure and the characteristics of near-surface sub-soil layers. Site-specific site effects due to geological conditions have been confirmed in recent earthquake events. Earthquake-induced damage has mainly occurred at accumulated soft soil layers under basins or along coasts and rivers. An alternative method has recently been proposed for evaluating regional seismic site effects and amplification factors using digital elevation models (DEM. High-quality DEMs at high resolutions may be employed to resolve finer-scale variations in topographic gradients and consequently, correlated site response parameters. Because there are many regions in South Korea lacking borehole datasets, which are insufficient for site classification only using borehole datasets, a DEM-based proxy for seismic zonation can be effective. Thus, in this study, geo-proxy-based site classification was proposed based on empirical correlations with site response parameters and conducted for regional zonation of seismic site effects to identify the amplification of characteristics in the western metropolitan areas of South Korea, depending on the site-specific geo-spatial conditions.

An AP endonuclease 1-DNA polymerase beta complex: theoretical prediction of interacting surfaces.

Directory of Open Access Journals (Sweden)

Alexej Abyzov

2008-04-01

Full Text Available Abasic (AP sites in DNA arise through both endogenous and exogenous mechanisms. Since AP sites can prevent replication and transcription, the cell contains systems for their identification and repair. AP endonuclease (APEX1 cleaves the phosphodiester backbone 5' to the AP site. The cleavage, a key step in the base excision repair pathway, is followed by nucleotide insertion and removal of the downstream deoxyribose moiety, performed most often by DNA polymerase beta (pol-beta. While yeast two-hybrid studies and electrophoretic mobility shift assays provide evidence for interaction of APEX1 and pol-beta, the specifics remain obscure. We describe a theoretical study designed to predict detailed interacting surfaces between APEX1 and pol-beta based on published co-crystal structures of each enzyme bound to DNA. Several potentially interacting complexes were identified by sliding the protein molecules along DNA: two with pol-beta located downstream of APEX1 (3' to the damaged site and three with pol-beta located upstream of APEX1 (5' to the damaged site. Molecular dynamics (MD simulations, ensuring geometrical complementarity of interfaces, enabled us to predict interacting residues and calculate binding energies, which in two cases were sufficient (approximately -10.0 kcal/mol to form a stable complex and in one case a weakly interacting complex. Analysis of interface behavior during MD simulation and visual inspection of interfaces allowed us to conclude that complexes with pol-beta at the 3'-side of APEX1 are those most likely to occur in vivo. Additional multiple sequence analyses of APEX1 and pol-beta in related organisms identified a set of correlated mutations of specific residues at the predicted interfaces. Based on these results, we propose that pol-beta in the open or closed conformation interacts and makes a stable interface with APEX1 bound to a cleaved abasic site on the 3' side. The method described here can be used for analysis in

Tannic acid and chromic chloride-induced binding of protein to red cells: a preliminary study of possible binding sites and reaction mechanisms.

Science.gov (United States)

Hunt, A F; Reed, M I

1990-07-01

The binding mechanisms and binding sites involved in the tannic acid and chromic chloride-induced binding of protein to red cells were investigated using the binding of IgA paraprotein to red cells as model systems. Inhibition studies of these model systems using amino acid homopolymers and compounds (common as red cell membrane constituents) suggest that the mechanisms involved are similar to those proposed for the conversion of hide or skin collagen to leather, as in commercial tanning. These studies also suggest that tannic acid-induced binding of IgA paraprotein to red cells involves the amino acid residues of L-arginine, L-lysine, L-histidine, and L-proline analogous to tanning with phenolic plant extracts. The amino acid residues of L-aspartate, L-glutamate and L-asparagine are involved in a similar manner in chronic chloride-induced binding of protein to red cells.

Pinpointing Phosphorylation Sites: Quantitative Filtering and a Novel Site-specific x-Ion Fragment

DEFF Research Database (Denmark)

Kelstrup, Christian D; Hekmat, Omid; Francavilla, Chiara

2011-01-01

Phosphoproteomics deals with the identification and quantification of thousands of phosphopeptides. Localizing the phosphorylation site is however much more difficult than establishing the identity of a phosphorylated peptide. Further, recent findings have raised doubts of the validity of the site......-phase phosphate rearrangement reactions during collision-induced dissociation (CID) and used these spectra to devise a quantitative filter that by comparing signal intensities of putative phosphorylated fragment ions with their nonphosphorylated counterparts allowed us to accurately pinpoint which fragment ions...... contain a phosphorylated residue and which ones do not. We also evaluated higher-energy collisional dissociation (HCD) and found this to be an accurate method for correct phosphorylation site localization with no gas-phase rearrangements observed above noise level. Analyzing a large set of HCD spectra...

Genome-wide identification of hypoxia-inducible factor-1 and -2 binding sites in hypoxic human macrophages alternatively activated by IL-10.

Science.gov (United States)

Tausendschön, Michaela; Rehli, Michael; Dehne, Nathalie; Schmidl, Christian; Döring, Claudia; Hansmann, Martin-Leo; Brüne, Bernhard

2015-01-01

Macrophages (MΦ) often accumulate in hypoxic areas, where they significantly influence disease progression. Anti-inflammatory cytokines, such as IL-10, generate alternatively activated macrophages that support tumor growth. To understand how alternative activation affects the transcriptional profile of hypoxic macrophages, we globally mapped binding sites of hypoxia-inducible factor (HIF)-1α and HIF-2α in primary human monocyte-derived macrophages prestimulated with IL-10. 713 HIF-1 and 795 HIF-2 binding sites were identified under hypoxia. Pretreatment with IL-10 altered the binding pattern, with 120 new HIF-1 and 188 new HIF-2 binding sites emerging. HIF-1 binding was most prominent in promoters, while HIF-2 binding was more abundant in enhancer regions. Comparison of ChIP-seq data obtained in other cells revealed a highly cell type specific binding of HIF. In MΦ HIF binding occurred preferentially in already active enhancers or promoters. To assess the roles of HIF on gene expression, primary human macrophages were treated with siRNA against HIF-1α or HIF-2α, followed by genome-wide gene expression analysis. Comparing mRNA expression to the HIF binding profile revealed a significant enrichment of hypoxia-inducible genes previously identified by ChIP-seq. Analysis of gene expression under hypoxia alone and hypoxia/IL-10 showed the enhanced induction of a set of genes including PLOD2 and SLC2A3, while another group including KDM3A and ADM remained unaffected or was reduced by IL-10. Taken together IL-10 influences the DNA binding pattern of HIF and the level of gene induction. Copyright © 2014 Elsevier B.V. All rights reserved.

ZnO nanopowder induced light scattering for improved visualization of emission sites in carbon nanotube films and arrays

Science.gov (United States)

Meško, Marcel; Ou, Qiongrong; Matsuda, Takafumi; Ishikawa, Tomokazu; Veis, Martin; Antoš, Roman; Ogino, Akihisa; Nagatsu, Masaaki

2009-06-01

We report on ZnO nanopowder induced light scattering for improved visualization of emission sites in carbon nanotube films and arrays. We observed a significant reduction of the internal multiple light scattering phenomena, which are characteristic for ZnO micropowders. The microsized grains of the commercially available ZnO:Zn (P 15) were reduced to the nanometre scale by pulsed laser ablation at an oxygen ambient pressure of 10 kPa. Our investigations show no crystalline change and no shift of the broad green emission peak at 500 nm for the ZnO nanopowder. For the application in field emission displays, we demonstrate the possibility of achieving cathodoluminescence with a fine pitch size of 100 µm of the patterned pixels without requiring additional electron beam focusing and without a black matrix. Moreover, the presented results show the feasibility of employing ZnO nanopowder as a detection material for the phosphorus screen method, which is able to localize emission sites of carbon nanotube films and arrays with an accuracy comparable to scanning anode field emission microscopy.

ZnO nanopowder induced light scattering for improved visualization of emission sites in carbon nanotube films and arrays

International Nuclear Information System (INIS)

Mesko, Marcel; Ou Qiongrong; Matsuda, Takafumi; Ishikawa, Tomokazu; Ogino, Akihisa; Nagatsu, Masaaki; Veis, Martin; Antos, Roman

2009-01-01

We report on ZnO nanopowder induced light scattering for improved visualization of emission sites in carbon nanotube films and arrays. We observed a significant reduction of the internal multiple light scattering phenomena, which are characteristic for ZnO micropowders. The microsized grains of the commercially available ZnO:Zn (P 15) were reduced to the nanometre scale by pulsed laser ablation at an oxygen ambient pressure of 10 kPa. Our investigations show no crystalline change and no shift of the broad green emission peak at 500 nm for the ZnO nanopowder. For the application in field emission displays, we demonstrate the possibility of achieving cathodoluminescence with a fine pitch size of 100 μm of the patterned pixels without requiring additional electron beam focusing and without a black matrix. Moreover, the presented results show the feasibility of employing ZnO nanopowder as a detection material for the phosphorus screen method, which is able to localize emission sites of carbon nanotube films and arrays with an accuracy comparable to scanning anode field emission microscopy.

Siting of research reactors

International Nuclear Information System (INIS)

1987-01-01

The purpose of this document is to develop criteria for siting and the site-related design basis for research reactors. The concepts presented in this document are intended as recommendations for new reactors and are not suggested for backfitting purposes for facilities already in existence. In siting research reactors serious consideration is given to minimizing the effects of the site on the reactor and the reactor on the site and the potential impact of the reactor on the environment. In this document guidance is first provided on the evaluation of the radiological impact of the installation under normal reactor operation and accident conditions. A classification of research reactors in groups is then proposed, together with a different approach for each group, to take into account the relevant safety problems associated with facilities of different characteristics. Guidance is also provided for both extreme natural events and for man-induced external events which could affect the safe operation of the reactor. Extreme natural events include earthquakes, flooding for river or coastal sites and extreme meteorological phenomena. The feasibility of emergency planning is finally considered for each group of reactors

Alkyladenine DNA glycosylase (Aag) in somatic hypermutation and class switch recombination.

Science.gov (United States)

Longerich, Simonne; Meira, Lisiane; Shah, Dharini; Samson, Leona D; Storb, Ursula

2007-12-01

Somatic hypermutation (SHM) and class switch recombination (CSR) of immunoglobulin (Ig) genes require the cytosine deaminase AID, which deaminates cytosine to uracil in Ig gene DNA. Paradoxically, proteins involved normally in error-free base excision repair and mismatch repair, seem to be co-opted to facilitate SHM and CSR, by recruiting error-prone translesion polymerases to DNA sequences containing deoxy-uracils created by AID. Major evidence supports at least one mechanism whereby the uracil glycosylase Ung removes AID-generated uracils creating abasic sites which may be used either as uninformative templates for DNA synthesis, or processed to nicks and gaps that prime error-prone DNA synthesis. We investigated the possibility that deamination at adenines also initiates SHM. Adenosine deamination would generate hypoxanthine (Hx), a substrate for the alkyladenine DNA glycosylase (Aag). Aag would generate abasic sites which then are subject to error-prone repair as above for AID-deaminated cytosine processed by Ung. If the action of an adenosine deaminase followed by Aag were responsible for significant numbers of mutations at A, we would find a preponderance of A:T>G:C transition mutations during SHM in an Aag deleted background. However, this was not observed and we found that the frequencies of SHM and CSR were not significantly altered in Aag-/- mice. Paradoxically, we found that Aag is expressed in B lymphocytes undergoing SHM and CSR and that its activity is upregulated in activated B cells. Moreover, we did find a statistically significant, albeit low increase of T:A>C:G transition mutations in Aag-/- animals, suggesting that Aag may be involved in creating the SHM A>T bias seen in wild type mice.

Drought-induced legacy effects in wood growth across the Eastern and Midwestern U.S. are mediated by site climate, tree age, and drought sensitivity

Science.gov (United States)

Kannenberg, S.; Maxwell, J. T.; Pederson, N.; D'Orangeville, L.; Phillips, R.

2017-12-01

While it is widely known that drought reduces carbon (C) uptake in temperate forests, tree growth can also remain stagnant post-drought despite favorable climatic conditions. While such "legacy effects" are well established, the degree to which these effects depend on species identity or variability in site conditions is poorly quantified. We sought to uncover how site, species, climate, and tree age interact to affect the presence and magnitude of legacy effects in temperate trees following drought. To do this, we assembled dendrochronological records of 18 common species across 94 sites in Eastern and Midwestern U.S. forests and quantified drought-induced changes in wood growth in the year of the drought (hereafter "drought sensitivity") and the years after the drought (i.e., legacy effects). We predicted that species particularly prone to hydraulic damage (e.g., oaks) would have the least drought sensitivity yet experience larger legacy effects, and that this effect would be exacerbated at arid sites. Across all species and sites, wood growth was reduced by 14% in the year of the drought and by 7% post-drought. Surprisingly, legacy effects were smaller for oak species and larger across species known to be more drought sensitive (e.g. tulip poplar, maple, birch). As a result, we observed a positive relationship between a species' drought sensitivity and that species' legacy effect. These legacy effects were similar in size across a range of drought severities. Surprisingly, legacy effects were smaller in more arid sites - contrary to previous investigations in dryland ecosystems - perhaps indicating the role of adaptation in mediating a tree's recovery from drought. In addition, many species actually decreased the size of their legacy effects as they aged, despite no change in drought responses. Our results run contrary to our predictions, as species with the greatest drought sensitivity had the least ability to recover, and that younger mesic forests- not arid

An immunochemical approach to the study of DNA damage and repair

International Nuclear Information System (INIS)

Wallace, S.S.; Erlanger, B.F.

1992-05-01

The overall objective of this project has been to develop immunochemical methods to quantitate unique DNA base damages in order to facilitate studies on radiation-induced damage production and repair. Specifically, we have been using antibodies raised to damaged bases to quantitate unique lesions in model systems in order to evaluate their potential biological consequences. Our approach has been to synthesize modified nucleotides or nucleosides, conjugate them to protein carriers, and use the conjugates as immunogens in rabbits or to prepare monoclonal antibodies. We have been studying damages that are stable radiolysis products found in X-irradiated DNA and thus of potential biological consequence. Our aim is to build an in vitro and in vivo data base on the interactions between model DNA lesions and such cellular enzymes as DNA polymerases and repair endonucleases. Initial studies have focused on pyrimidine ring saturation products (thymine glycol.and dihydrothymine), products resulting from ring fragmentation or base loss (urea, Β-ureidoisobutyric acid, abasic sites), 7-hydro-8-oxopurines, and more recently, cytosine radiolysis products. These modified bases serve as useful models for examining the potential lethal and/or mutagenic (carcinogenic) effects of the products of DNA radiolysis

Cold-Induced Thermogenesis and Inflammation-Associated Cold-Seeking Behavior Are Represented by Different Dorsomedial Hypothalamic Sites: A Three-Dimensional Functional Topography Study in Conscious Rats.

Science.gov (United States)

Wanner, Samuel P; Almeida, M Camila; Shimansky, Yury P; Oliveira, Daniela L; Eales, Justin R; Coimbra, Cândido C; Romanovsky, Andrej A

2017-07-19

In the past, we showed that large electrolytic lesions of the dorsomedial hypothalamus (DMH) promoted hypothermia in cold-exposed restrained rats, but attenuated hypothermia in rats challenged with a high dose of bacterial lipopolysaccharide (LPS) in a thermogradient apparatus. The goal of this study was to identify the thermoeffector mechanisms and DMH representation of the two phenomena and thus to understand how the same lesion could produce two opposite effects on body temperature. We found that the permissive effect of large electrolytic DMH lesions on cold-induced hypothermia was due to suppressed thermogenesis. DMH-lesioned rats also could not develop fever autonomically: they did not increase thermogenesis in response to a low, pyrogenic dose of LPS (10 μg/kg, i.v.). In contrast, changes in thermogenesis were uninvolved in the attenuation of the hypothermic response to a high, shock-inducing dose of LPS (5000 μg/kg, i.v.); this attenuation was due to a blockade of cold-seeking behavior. To compile DMH maps for the autonomic cold defense and for the cold-seeking response to LPS, we studied rats with small thermal lesions in different parts of the DMH. Cold thermogenesis had the highest representation in the dorsal hypothalamic area. Cold seeking was represented by a site at the ventral border of the dorsomedial nucleus. Because LPS causes both fever and hypothermia, we originally thought that the DMH contained a single thermoregulatory site that worked as a fever-hypothermia switch. Instead, we have found two separate sites: one that drives thermogenesis and the other, previously unknown, that drives inflammation-associated cold seeking. SIGNIFICANCE STATEMENT Cold-seeking behavior is a life-saving response that occurs in severe systemic inflammation. We studied this behavior in rats with lesions in the dorsomedial hypothalamus (DMH) challenged with a shock-inducing dose of bacterial endotoxin. We built functional maps of the DMH and found the strongest

Site-specific induction of nuclear anomalies (apoptotic bodies and micronuclei) by carcinogens in mice

International Nuclear Information System (INIS)

Ronen, A.; Heddle, J.A.

1984-01-01

The usefulness of nuclear anomalies (NA) as a short-term test for indication of carcinogens in the mouse colon has been suggested previously by experiments in which colon-specific carcinogens induced NA in the colon, whereas non-colon carcinogens were, in general, impotent in that organ. We have extended this work to other sites in the digestive tract of female C57BL/6 mice treated with gamma-rays, 1,2-dimethylhydrazine dihydrochloride, or N-methylnitrosourea. Each agent induced NA at all of the sites examined. The frequency of NA at different times after treatment depended upon both the agent used and the site examined. 1,2-Dimethylhydrazine dihydrochloride (which is known to induce tumors predominantly in the colon) induces NA with the highest efficiency (relative to gamma-rays) in the descending colon. N-Methylnitrosourea (which induces tumors mainly in the forestomach) induces NA with the highest efficiency in the forestomach. These results further support the usefulness of the assay in that the frequency of NA produced at the various sites by 1,2-dimethylhydrazine dihydrochloride and N-methylnitrosourea correlates with that found in the carcinogenicity studies

Evaluation of ground deformations induced by the 1999 Kocaeli earthquake (Turkey) at selected sites on shorelines

Science.gov (United States)

Aydan, Ömer; Ulusay, Reşat; Atak, Veysel Okan

2008-03-01

The Kocaeli earthquake ( M w = 7.4) of 17 August 1999 occurred in the Eastern Marmara Region of Turkey along the North Anadolu Fault and resulted in a very serious loss of life and property. One of the most important geotechnical issues of this event was the permanent ground deformations because of both liquefaction and faulting. These deformations occurred particularly along the southern shores of İzmit Bay and Sapanca Lake between the cities of Yalova and Adapazarı in the west and east, respectively. In this study, three sites founded on delta fans, namely Değirmendere Nose, Yeniköy tea garden at Seymen on the coast of İzmit Bay, and Vakıf Hotel site on the coast of Sapanca Lake were selected as typical cases. The main causes of the ground deformations at these sites were then investigated. Geotechnical characterization of the ground, derivation of displacement vectors from the pre- and post-earthquake aerial photographs, liquefaction assessments based on field performance data, and analyses carried out using the sliding body method have been fundamental in this study. The displacement vectors determined from photogrammetric evaluations conducted at Değirmendere and Seymen showed a combined movement of faulting and liquefaction. But except the movements in the close vicinity of shorelines, the dominant factor in this movement was faulting. The results obtained from the analyses suggested that the ground failure at Değirmendere was a submarine landslide mainly because of earthquake shaking rather than liquefaction. On the other hand, the ground failures at the Yeniköy tea garden on the coast of Seymen and the hotel area in Sapanca town resulted from liquefaction-induced lateral spreading. It was also obtained that the ground deformations estimated from the sliding body method were quite close to those measured by aerial photogrammetry technique.

Topic I: Induced changes in hydrology at low-level radioactive waste repository sites: A section in Safe disposal of radionuclides in low-level radioactive-waste repository sites; Low-level radioactive-waste disposal workshop, U.S. Geological Survey, July 11-16, 1987, Big Bear Lake, Calif., Proceedings (Circular 1036)

Science.gov (United States)

Prudic, David E.; Dennehy, Kevin F.; Bedinger, Marion S.; Stevens, Peter R.

1990-01-01

Engineering practices, including the excavation of trenches, placement of waste, nature of waste forms, backfilling procedures and materials, and trench-cover construction and materials at low-level radioactive-waste repository sites greatly affect the geohydrology of the sites. Engineering practices are dominant factors in eventual stability and isolation of the waste. The papers presented relating to Topic I were discussions of the hydrogeologic setting at existing low-level radioactive-waste repository sites and changes in the hydrology induced by site operations. Papers summarizing detailed studies presented at this workshop include those at sites near Sheffield, Ill.; Oak Ridge National Laboratory, Tenn.; West Valley, N.Y.; Maxey Flats, Ky.; Barnwell, S.C.; and Beatty, Nev. 

Mutation of the inhibitory ethanol site in GABAA ρ1 receptors promotes tolerance to ethanol-induced motor incoordination.

Science.gov (United States)

Blednov, Yuri A; Borghese, Cecilia M; Ruiz, Carlos I; Cullins, Madeline A; Da Costa, Adriana; Osterndorff-Kahanek, Elizabeth A; Homanics, Gregg E; Harris, R Adron

2017-09-01

Genes encoding the ρ1/2 subunits of GABA A receptors have been associated with alcohol (ethanol) dependence in humans, and ρ1 was also shown to regulate some of the behavioral effects of ethanol in animal models. Ethanol inhibits GABA-mediated responses in wild-type (WT) ρ1, but not ρ1(T6'Y) mutant receptors expressed in Xenopus laevis oocytes, indicating the presence of an inhibitory site for ethanol in the second transmembrane helix. In this study, we found that ρ1(T6'Y) receptors expressed in oocytes display overall normal responses to GABA, the endogenous GABA modulator (zinc), and partial agonists (β-alanine and taurine). We generated ρ1 (T6'Y) knockin (KI) mice using CRISPR/Cas9 to test the behavioral importance of the inhibitory actions of ethanol on this receptor. Both ρ1 KI and knockout (KO) mice showed faster recovery from acute ethanol-induced motor incoordination compared to WT mice. Both KI and KO mutant strains also showed increased tolerance to motor impairment produced by ethanol. The KI mice did not differ from WT mice in other behavioral actions, including ethanol intake and preference, conditioned taste aversion to ethanol, and duration of ethanol-induced loss of righting reflex. WT and KI mice did not differ in levels of ρ1 or ρ2 mRNA in cerebellum or in ethanol clearance. Our findings indicate that the inhibitory site for ethanol in GABA A ρ1 receptors regulates acute functional tolerance to moderate ethanol intoxication. We note that low sensitivity to alcohol intoxication has been linked to risk for development of alcohol dependence in humans. Copyright © 2017 Elsevier Ltd. All rights reserved.

Structure of the Dispase Autolysis-inducing Protein from Streptomyces mobaraensis and Glutamine Cross-linking Sites for Transglutaminase.

Science.gov (United States)

Fiebig, David; Schmelz, Stefan; Zindel, Stephan; Ehret, Vera; Beck, Jan; Ebenig, Aileen; Ehret, Marina; Fröls, Sabrina; Pfeifer, Felicitas; Kolmar, Harald; Fuchsbauer, Hans-Lothar; Scrima, Andrea

2016-09-23

Transglutaminase from Streptomyces mobaraensis (MTG) is an important enzyme for cross-linking and modifying proteins. An intrinsic substrate of MTG is the dispase autolysis-inducing protein (DAIP). The amino acid sequence of DAIP contains 5 potential glutamines and 10 lysines for MTG-mediated cross-linking. The aim of the study was to determine the structure and glutamine cross-linking sites of the first physiological MTG substrate. A production procedure was established in Escherichia coli BL21 (DE3) to obtain high yields of recombinant DAIP. DAIP variants were prepared by replacing four of five glutamines for asparagines in various combinations via site-directed mutagenesis. Incorporation of biotin cadaverine revealed a preference of MTG for the DAIP glutamines in the order of Gln-39 ≫ Gln-298 > Gln-345 ∼ Gln-65 ≫ Gln-144. In the structure of DAIP the preferred glutamines do cluster at the top of the seven-bladed β-propeller. This suggests a targeted cross-linking of DAIP by MTG that may occur after self-assembly in the bacterial cell wall. Based on our biochemical and structural data of the first physiological MTG substrate, we further provide novel insight into determinants of MTG-mediated modification, specificity, and efficiency. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Site-specific genome editing for correction of induced pluripotent stem cells derived from dominant dystrophic epidermolysis bullosa.

Science.gov (United States)

Shinkuma, Satoru; Guo, Zongyou; Christiano, Angela M

2016-05-17

Genome editing with engineered site-specific endonucleases involves nonhomologous end-joining, leading to reading frame disruption. The approach is applicable to dominant negative disorders, which can be treated simply by knocking out the mutant allele, while leaving the normal allele intact. We applied this strategy to dominant dystrophic epidermolysis bullosa (DDEB), which is caused by a dominant negative mutation in the COL7A1 gene encoding type VII collagen (COL7). We performed genome editing with TALENs and CRISPR/Cas9 targeting the mutation, c.8068_8084delinsGA. We then cotransfected Cas9 and guide RNA expression vectors expressed with GFP and DsRed, respectively, into induced pluripotent stem cells (iPSCs) generated from DDEB fibroblasts. After sorting, 90% of the iPSCs were edited, and we selected four gene-edited iPSC lines for further study. These iPSCs were differentiated into keratinocytes and fibroblasts secreting COL7. RT-PCR and Western blot analyses revealed gene-edited COL7 with frameshift mutations degraded at the protein level. In addition, we confirmed that the gene-edited truncated COL7 could neither associate with normal COL7 nor undergo triple helix formation. Our data establish the feasibility of mutation site-specific genome editing in dominant negative disorders.

Cluster analysis for the probability of DSB site induced by electron tracks

Energy Technology Data Exchange (ETDEWEB)

Yoshii, Y. [Biological Research, Education and Instrumentation Center, Sapporo Medical University, Sapporo 060-8556 (Japan); Graduate School of Health Sciences, Hokkaido University, Sapporo 060-0812 (Japan); Sasaki, K. [Faculty of Health Sciences, Hokkaido University of Science, Sapporo 006-8585 (Japan); Matsuya, Y. [Graduate School of Health Sciences, Hokkaido University, Sapporo 060-0812 (Japan); Date, H., E-mail: date@hs.hokudai.ac.jp [Faculty of Health Sciences, Hokkaido University, Sapporo 060-0812 (Japan)

2015-05-01

To clarify the influence of bio-cells exposed to ionizing radiations, the densely populated pattern of the ionization in the cell nucleus is of importance because it governs the extent of DNA damage which may lead to cell lethality. In this study, we have conducted a cluster analysis of ionization and excitation events to estimate the number of double-strand breaks (DSBs) induced by electron tracks. A Monte Carlo simulation for electrons in liquid water was performed to determine the spatial location of the ionization and excitation events. The events were divided into clusters by using the density-based spatial clustering of applications with noise (DBSCAN) algorithm. The algorithm enables us to sort out the events into the groups (clusters) in which a minimum number of neighboring events are contained within a given radius. For evaluating the number of DSBs in the extracted clusters, we have introduced an aggregation index (AI). The computational results show that a sub-keV electron produces DSBs in a dense formation more effectively than higher energy electrons. The root-mean square radius (RMSR) of the cluster size is below 5 nm, which is smaller than the chromatin fiber thickness. It was found that this size of clustering events has a high possibility to cause lesions in DNA within the chromatin fiber site.

Chemically induced DNA hypomethylation in breast carcinoma cells detected by the amplification of intermethylated sites

International Nuclear Information System (INIS)

Sadikovic, Bekim; Haines, Thomas R; Butcher, Darci T; Rodenhiser, David I

2004-01-01

Compromised patterns of gene expression result in genomic instability, altered patterns of gene expression and tumour formation. Specifically, aberrant DNA hypermethylation in gene promoter regions leads to gene silencing, whereas global hypomethylation events can result in chromosomal instability and oncogene activation. Potential links exist between environmental agents and DNA methylation, but the destabilizing effects of environmental exposures on the DNA methylation machinery are not understood within the context of breast cancer aetiology. We assessed genome-wide changes in methylation patterns using a unique methylation profiling technique called amplification of intermethylated sites (AIMS). This method generates easily readable fingerprints that represent the investigated cell line's methylation profile, based on the differential cleavage of DNA with methylation-specific isoschisomeric restriction endonucleases. We validated this approach by demonstrating both unique and reoccurring sites of genomic hypomethylation in four breast carcinoma cell lines treated with the cytosine analogue 5-azacytidine. Comparison of treated with control samples revealed individual bands that exhibited methylation changes, and these bands were excized and cloned, and the precise genomic location individually identified. In most cases, these regions of hypomethylation coincided with susceptible target regions previously associated with chromosome breakage, rearrangement and gene amplification. Similarly, we observed that acute benzopyrene exposure is associated with altered methylation patterns in these cell lines. These results reinforce the link between environmental exposures, DNA methylation and breast cancer, and support a role for AIMS as a rapid, affordable screening method to identify environmentally induced DNA methylation changes that occur in tumourigenesis

Seismically-induced soil amplification at the DOE Paducah Gaseous Diffusion Plant site

International Nuclear Information System (INIS)

Sykora, D.W.; Haynes, M.E.

1991-01-01

A site-specific earthquake site response (soil amplification) study is being conducted for the Department of Energy (DOE), Paducah Gaseous Diffusion Plant (PGDP). This study is pursuant to an upgraded Final Safety Analysis Report in accordance with requirements specified by DOE. The seismic hazard at PGDP is dominated by the New Madrid Seismic Zone. Site-specific synthetic earthquake records developed by others were applied independently to four soil columns with heights above baserock of about 325 ft. The results for the 1000-year earthquake event indicate that the site period is between 1.0 and 1.5 sec. Incident shear waves are amplified at periods of motion greater than 0.15 sec. The peak free-field horizontal acceleration, occurring at very low periods, is 0.28 g. 13 refs., 13 figs

Seismically-induced soil amplification at the DOE Paducah Gaseous Diffusion Plant Site

International Nuclear Information System (INIS)

Sykora, D.W.; Hynes, M.E.; Brock, W.R.; Hunt, R.J.; Shaffer, K.E.

1991-01-01

A site-specific earthquake site response (soil amplification) study is being conducted for the Department of Energy (DOE), Paducah Gaseous Diffusion Plant (PGDP). This study is pursuant to an upgraded Final Safety Analysis Report in accordance with requirements specified by DOE. The seismic hazard at PGDP is dominated by the New Madrid Seismic Zone. Site-specific synthetic earthquake records developed by others were applied independently to four soil columns with heights above baserock of about 325 ft. The results for the 1000-year earthquake event indicate that the site period is between 1.0 and 1.5 sec. Incident shear waves are strongly amplified at periods of motion greater than 0.3 sec. The peak free-field horizontal acceleration, occurring at very low periods, is 0.28 g

An investigation of Laser Induced Breakdown Spectroscopy for use as a control in the laser removal of rock from fossils found at the Malapa hominin site, South Africa

CSIR Research Space (South Africa)

Roberts, DE

2012-07-01

Full Text Available Laser Induced Breakdown Spectroscopy (LIBS) was used to study the spectra from fossils and surrounding rock recovered from the Cradle of Mankind site at Malapa, South Africa. The objective was to find a suitable spectral line(s), specific to fossils...

Tourism-Induced Livelihood Changes at Mount Sanqingshan World Heritage Site, China

Science.gov (United States)

Su, Ming Ming; Wall, Geoffrey; Xu, Kejian

2016-05-01

Although tourism has the potential to improve the wellbeing of residents, it may also disrupt livelihood systems, social processes, and cultural traditions. The livelihood changes at three rural villages at Mount Sanqingshan World Heritage Site, China, are assessed to determine the extent to which tourism strategies are contributing to local livelihoods. A sustainable livelihood framework is adopted to guide the analysis. The three villages exhibit different development patterns due to institutional, organizational, and location factors. New strategies involving tourism were constructed and incorporated into the traditional livelihood systems and they resulted in different outcomes for residents of different villages. Village location, including the relationship to the site tourism plan, affected the implications for rural livelihoods. High dependence on tourism as the single livelihood option can reduce sustainability. Practical implications are suggested to enhance livelihood sustainability at such rural heritage tourism sites.

Tourism-Induced Livelihood Changes at Mount Sanqingshan World Heritage Site, China.

Science.gov (United States)

Su, Ming Ming; Wall, Geoffrey; Xu, Kejian

2016-05-01

Although tourism has the potential to improve the wellbeing of residents, it may also disrupt livelihood systems, social processes, and cultural traditions. The livelihood changes at three rural villages at Mount Sanqingshan World Heritage Site, China, are assessed to determine the extent to which tourism strategies are contributing to local livelihoods. A sustainable livelihood framework is adopted to guide the analysis. The three villages exhibit different development patterns due to institutional, organizational, and location factors. New strategies involving tourism were constructed and incorporated into the traditional livelihood systems and they resulted in different outcomes for residents of different villages. Village location, including the relationship to the site tourism plan, affected the implications for rural livelihoods. High dependence on tourism as the single livelihood option can reduce sustainability. Practical implications are suggested to enhance livelihood sustainability at such rural heritage tourism sites.

Procedural justice in wind facility siting: Recommendations for state-led siting processes

International Nuclear Information System (INIS)

Ottinger, Gwen; Hargrave, Timothy J.; Hopson, Eric

2014-01-01

Evidence suggests that state control of wind facility siting decisions fosters new project development more effectively than local control, yet the literature suggests that affected citizens tend to be more fairly represented in local siting processes. We argue that successful renewable energy policy must satisfy both the need for new project development and the obligation to procedural justice. To suggest how it can do so, we analyze existing state- and county-level siting processes in Washington state, finding that both fall short on measures of procedural justice. To overcome this limitation and address the tension between procedural justice and project development, we then propose a collaborative governance approach to wind facility siting, in which state governments retain ultimate authority over permitting decisions but encourage and support local-level deliberations as the primary means of making those decisions. Such an approach, we argue, would be more just, facilitate wind development by addressing community concerns constructively and result in better projects through the input of diverse stakeholders. - Highlights: • States have made wind energy development a priority. • Local opposition to new projects could hinder future wind energy development. • Procedural justice is necessary to resolve local issues and ensure timely wind facility siting. • Both state- and county-led siting processes fall short with respect to criteria for procedural justice, though local processes have some advantages. • States could instead induce counties, developers to engage in deliberation

Lipopolysaccharide-induced inhibition of transcription of tlr4 in vitro is reversed by dexamethasone and correlates with presence of conserved NFκB binding sites

Energy Technology Data Exchange (ETDEWEB)

Bonin, Camila P., E-mail: mila_bonin@yahoo.com.br [Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo 05508-900 (Brazil); Baccarin, Raquel Y.A., E-mail: baccarin@usp.br [Department of Clinics, School of Veterinary Medicine, University of São Paulo, São Paulo 05508-900 (Brazil); Nostell, Katarina, E-mail: katarina.nostell@slu.se [Department of Clinical Sciences, Swedish University of Agricultural Sciences, Box 7054, 750 07 Uppsala (Sweden); Nahum, Laila A., E-mail: laila@nahum.com.br [Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, Belo Horizonte 30190-002 (Brazil); Faculdade Infórium de Tecnologia, Belo Horizonte 30130-180 (Brazil); Fossum, Caroline, E-mail: caroline.fossum@bvf.slu.se [Department of Biomedicine and Veterinary Public Health, Section for Immunology, Swedish University of Agricultural Sciences, BMC, Box 588, SE 751 23 Uppsala (Sweden); Camargo, Maristela M. de, E-mail: mmcamar@usp.br [Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo 05508-900 (Brazil)

2013-03-08

Highlights: ► Chimpanzees, horses and humans have regions of similarity on TLR4 and MD2 promoters. ► Rodents have few regions of similarity on TLR4 promoter when compared to primates. ► Conserved NFkB binding sites were found in the promoters of TLR4 and MD2. ► LPS-induced inhibition of TLR4 transcription is reversed by dexamethasone. ► LPS-induced transcription of MD2 is inhibited by dexamethasone. -- Abstract: Engagement of Toll-like receptor 4 (TLR4) by lipopolysaccharide (LPS) is a master trigger of the deleterious effects of septic shock. Horses and humans are considered the most sensitive species to septic shock, but the mechanisms explaining these phenomena remain elusive. Analysis of tlr4 promoters revealed high similarity among LPS-sensitive species (human, chimpanzee, and horse) and low similarity with LPS-resistant species (mouse and rat). Four conserved nuclear factor kappa B (NFκB) binding sites were found in the tlr4 promoter and two in the md2 promoter sequences that are likely to be targets for dexamethasone regulation. In vitro treatment of equine peripheral blood mononuclear cells (eqPBMC) with LPS decreased transcripts of tlr4 and increased transcription of md2 (myeloid differentiation factor 2) and cd14 (cluster of differentiation 14). Treatment with dexamethasone rescued transcription of tlr4 after LPS inhibition. LPS-induced transcription of md2 was inhibited in the presence of dexamethasone. Dexamethasone alone did not affect transcription of tlr4 and md2.

Determination and analysis of site-specific 125I decay-induced DNA double-strand break end-group structures.

Science.gov (United States)

Datta, Kamal; Weinfeld, Michael; Neumann, Ronald D; Winters, Thomas A

2007-02-01

End groups contribute to the structural complexity of radiation-induced DNA double-strand breaks (DSBs). As such, end-group structures may affect a cell's ability to repair DSBs. The 3'-end groups of strand breaks caused by gamma radiation, or oxidative processes, under oxygenated aqueous conditions have been shown to be distributed primarily between 3'-phosphoglycolate and 3'-phosphate, with 5'-phosphate ends in both cases. In this study, end groups of the high-LET-like DSBs caused by 125I decay were investigated. Site-specific DNA double-strand breaks were produced in plasmid pTC27 in the presence or absence of 2 M DMSO by 125I-labeled triplex-forming oligonucleotide targeting. End-group structure was assessed enzymatically as a function of the DSB end to serve as a substrate for ligation and various forms of end labeling. Using this approach, we have demonstrated 3'-hydroxyl (3'-OH) and 3'-phosphate (3'-P) end groups and 5'-ends (> or = 42%) terminated by phosphate. A 32P postlabeling assay failed to detect 3'-phosphoglycolate in a restriction fragment terminated by the 125I-induced DNA double-strand break, and this is likely due to restricted oxygen diffusion during irradiation as a frozen aqueous solution. Even so, end-group structure and relative distribution varied as a function of the free radical scavenging capacity of the irradiation buffer.

Site-specific effects of apolipoprotein E expression on diet-induced obesity and white adipose tissue metabolic activation.

Science.gov (United States)

Hatziri, Aikaterini; Kalogeropoulou, Christina; Xepapadaki, Eva; Birli, Eleni; Karavia, Eleni A; Papakosta, Eugenia; Filou, Serafoula; Constantinou, Caterina; Kypreos, Kyriakos E

2018-02-01

Apolipoprotein E (APOE) has been strongly implicated in the development of diet induced obesity. In the present study, we investigated the contribution of brain and peripherally expressed human apolipoprotein E3 (APOE3), the most common human isoform, to diet induced obesity. In our studies APOE3 knock-in (Apoe3 knock-in ), Apoe-deficient (apoe -/- ) and brain-specific expressing APOE3 (Apoe3 brain ) mice were fed western-type diet for 12week and biochemical analyses were performed. Moreover, AAV-mediated gene transfer of APOE3 to apoe -/- mice was employed, as a means to achieve APOE3 expression selectively in periphery, since peripherally expressed APOE does not cross blood brain barrier (BBB) or blood-cerebrospinal fluid barrier (BCSFB). Our data suggest a bimodal role of APOE3 in visceral white adipose tissue (WAT) mitochondrial metabolic activation that is highly dependent on its site of expression and independent of postprandial dietary lipid deposition. Our findings indicate that brain APOE3 expression is associated with a potent inhibition of visceral WAT mitochondrial oxidative phosphorylation, leading to significantly reduced substrate oxidation, increased fat accumulation and obesity. In contrast, peripherally expressed APOE3 is associated with a notable shift of substrate oxidation towards non-shivering thermogenesis in visceral WAT mitochondria, leading to resistance to obesity. Copyright © 2017 Elsevier B.V. All rights reserved.

Safety Review Services, Site Review Services and IRRS

International Nuclear Information System (INIS)

Yllera, Javier

2010-01-01

The selection and the evaluation of the site for a nuclear power plant are crucial parts of establishing a nuclear power programme and can be significantly affected by costs, public acceptance and safety considerations. Siting is the process of selecting a suitable site for a facility. This is area containing the plant, defined by a boundary and under effective control of the Plant Management. For safety related issues comparison within topics is generally quite straightforward. For example, sites with relatively higher seismic hazard would be penalized in comparison with those in more stable areas. The site for the NPP is generally chosen at a relatively ‘aseismic’ part of the country. This generally means that well known seismogenic sources are more than at least 50 kms from the site. The proposed sites for nuclear installations shall be examined with respect to the frequency and the severity of natural and human induced events and phenomena that could affect the safety of the installation. The Events unconnected with the operation of a facility or activity which could have an effect on the safety of the facility or activity. The relationship between the site and the design for the nuclear installation shall be examined to ensure that the radiological risk to the public and the environment arising from releases defined by the source terms is acceptably low. The Nuclear Regulatory Authority should issue a document that sets out the technical safety and security criteria against which the Site Permit Application for a new NPP will be reviewed. The objective of the Site Safety Review Services (SSRS) is provided upon request from a Member State. An independent review and assessment of the site and nuclear installation safety in relation to external natural and man induced hazards. This is to make recommendations on additional analysis or plant modifications to be carried out in order to comply with the IAEA Safety Standards and to enhance safety

Microwave-induced activation of additional active edge sites on the MoS2 surface for enhanced Hg0 capture

Science.gov (United States)

Zhao, Haitao; Mu, Xueliang; Yang, Gang; Zheng, Chengheng; Sun, Chenggong; Gao, Xiang; Wu, Tao

2017-10-01

In recent years, significant effort has been made in the development of novel materials for the removal of mercury from coal-derived flue gas. In this research, microwave irradiation was adopted to induce the creation of additional active sites on the MoS2 surface. The results showed that Hg0 capture efficiency of the adsorbent containing MoS2 nanosheets being microwave treated was as high as 97%, while the sample prepared via conventional method only showed an efficiency of 94% in its first 180 min testing. After the adsorbent was treated by microwave irradiation for 3 more times, its mercury removal efficiency was still noticeably higher than that of the sample prepared via conventional method. Characterization of surface structure of the MoS2 containing material together with DFT study further revealed that the (001) basal planes of MoS2 crystal structure were cracked into (100) edge planes (with an angle of approximately 75°) under microwave treatment, which subsequently resulted in the formation of additional active edge sites on the MoS2 surface and led to the improved performance on Hg0 capture.

DNA damage and radical reactions: Mechanistic aspects, formation in cells and repair studies

International Nuclear Information System (INIS)

Cadet, J.; Ravanat, J.L.; Carell, T.; Cellai, L.; Chatgilialoglu, Ch.; Gimisis, Th.; Miranda, M.; O'Neill, P.; Robert, M.

2008-01-01

Several examples of oxidative and reductive reactions of DNA components that lead to single and tandem modifications are discussed in this review. These include nucleophilic addition reactions of the one-electron oxidation-mediated guanine radical cation and the one-electron reduced intermediate of 8-bromo-purine 2'-de-oxy-ribo-nucleosides that give rise to either an oxidizing guanine radical or related 5',8-cyclo-purine nucleosides. In addition, mechanistic insights into the reductive pathways involved in the photolyase induced reversal of cyclo-buta-cli-pyrimidine and pyrimidine (6-4) pyrimidone photoproducts are provided. Evidence for the occurrence and validation in cellular DNA of (OH) · radical degradation pathways of guanine that have been established in model systems has been gained from the accurate measurement of degradation products. Relevant information on biochemical aspects of the repair of single and clustered oxidatively generated damage to DNA has been gained from detailed investigations that rely on the synthesis of suitable modified probes. Thus the preparation of stable carbocyclic derivatives of purine nucleoside containing defined sequence oligonucleotides has allowed detailed crystallographic studies of the recognition step of the base damage by enzymes implicated in the base excision repair (BER) pathway. Detailed insights are provided on the BER processing of non-double strand break bi-stranded clustered damage that may consist of base lesions, a single strand break or abasic sites and represent one of the main deleterious classes of radiation-induced DNA damage. (authors)

Lack of spontaneous and radiation-induced chromosome breakage at interstitial telomeric sites in murine scid cells.

Science.gov (United States)

Wong, H-P; Mozdarani, H; Finnegan, C; McIlrath, J; Bryant, P E; Slijepcevic, P

2004-01-01

Interstitial telomeric sites (ITSs) in chromosomes from DNA repair-proficient mammalian cells are sensitive to both spontaneous and radiation-induced chromosome breakage. Exact mechanisms of this chromosome breakage sensitivity are not known. To investigate factors that predispose ITSs to chromosome breakage we used murine scid cells. These cells lack functional DNA-PKcs, an enzyme involved in the repair of DNA double-strand breaks. Interestingly, our results revealed lack of both spontaneous and radiation-induced chromosome breakage at ITSs found in scid chromosomes. Therefore, it is possible that increased sensitivity of ITSs to chromosome breakage is associated with the functional DNA double-strand break repair machinery. To investigate if this is the case we used scid cells in which DNA-PKcs deficiency was corrected. Our results revealed complete disappearance of ITSs in scid cells with functional DNA-PKcs, presumably through chromosome breakage at ITSs, but their unchanged frequency in positive and negative control cells. Therefore, our results indicate that the functional DNA double-strand break machinery is required for elevated sensitivity of ITSs to chromosome breakage. Interestingly, we observed significant differences in mitotic chromosome condensation between scid cells and their counterparts with restored DNA-PKcs activity suggesting that lack of functional DNA-PKcs may cause a defect in chromatin organization. Increased condensation of mitotic chromosomes in the scid background was also confirmed in vivo. Therefore, our results indicate a previously unanticipated role of DNA-PKcs in chromatin organisation, which could contribute to the lack of ITS sensitivity to chromosome breakage in murine scid cells. Copyright 2003 S. Karger AG, Basel

Application of time domain induced polarization to the mapping of lithotypes in a landfill site

Directory of Open Access Journals (Sweden)

A. Gazoty

2012-06-01

Full Text Available A direct current (DC resistivity and time domain induced polarization (TDIP survey was undertaken at a decommissioned landfill site situated in Hørløkke, Denmark, for the purpose of mapping the waste deposits and to discriminate important geological units that control the hydrology of the surrounding area. It is known that both waste deposits and clay have clear signatures in TDIP data, making it possible to enhance the resolution of geological structures compared to DC surveys alone.

Four DC/TDIP profiles were carried out crossing the landfill, and another seven profiles in the surroundings provide a sufficiently dense coverage of the entire area. The whole dataset was inverted using a 1-D laterally constrained inversion scheme, recently implemented for TDIP data, in order to use the entire decay curves for reconstructing the electrical parameters of the soil in terms of the Cole-Cole polarization model.

Results show that it is possible to resolve both the geometry of the buried waste body and key geological structures. In particular, it was possible to find a silt/clay lens at depth that correlates with the flow direction of the pollution plume spreading out from the landfill and to map a shallow sandy layer rich in clay that likely has a strong influence on the hydrology of the site. This interpretation of the geophysical findings was constrained by borehole data, in terms of geology and gamma ray logging. The results of this study are important for the impact of the resolved geological units on the hydrology of the area, making it possible to construct more realistic scenarios of the variation of the pollution plume as a function of the climate change.

Relations between rainfall–runoff-induced erosion and aeolian deposition at archaeological sites in a semi-arid dam-controlled river corridor

Science.gov (United States)

Collins, Brian D.; Bedford, David; Corbett, Skye C.; Fairley, Helen C.; Cronkite-Ratcliff, Collin

2016-01-01

Process dynamics in fluvial-based dryland environments are highly complex with fluvial, aeolian, and alluvial processes all contributing to landscape change. When anthropogenic activities such as dam-building affect fluvial processes, the complexity in local response can be further increased by flood- and sediment-limiting flows. Understanding these complexities is key to predicting landscape behavior in drylands and has important scientific and management implications, including for studies related to paleoclimatology, landscape ecology evolution, and archaeological site context and preservation. Here we use multi-temporal LiDAR surveys, local weather data, and geomorphological observations to identify trends in site change throughout the 446-km-long semi-arid Colorado River corridor in Grand Canyon, Arizona, USA, where archaeological site degradation related to the effects of upstream dam operation is a concern. Using several site case studies, we show the range of landscape responses that might be expected from concomitant occurrence of dam-controlled fluvial sand bar deposition, aeolian sand transport, and rainfall-induced erosion. Empirical rainfall-erosion threshold analyses coupled with a numerical rainfall–runoff–soil erosion model indicate that infiltration-excess overland flow and gullying govern large-scale (centimeter- to decimeter-scale) landscape changes, but that aeolian deposition can in some cases mitigate gully erosion. Whereas threshold analyses identify the normalized rainfall intensity (defined as the ratio of rainfall intensity to hydraulic conductivity) as the primary factor governing hydrologic-driven erosion, assessment of false positives and false negatives in the dataset highlight topographic slope as the next most important parameter governing site response. Analysis of 4+ years of high resolution (four-minute) weather data and 75+ years of low resolution (daily) climate records indicates that dryland erosion is dependent on short

A novel human AP endonuclease with conserved zinc-finger-like motifs involved in DNA strand break responses

Science.gov (United States)

Kanno, Shin-ichiro; Kuzuoka, Hiroyuki; Sasao, Shigeru; Hong, Zehui; Lan, Li; Nakajima, Satoshi; Yasui, Akira

2007-01-01

DNA damage causes genome instability and cell death, but many of the cellular responses to DNA damage still remain elusive. We here report a human protein, PALF (PNK and APTX-like FHA protein), with an FHA (forkhead-associated) domain and novel zinc-finger-like CYR (cysteine–tyrosine–arginine) motifs that are involved in responses to DNA damage. We found that the CYR motif is widely distributed among DNA repair proteins of higher eukaryotes, and that PALF, as well as a Drosophila protein with tandem CYR motifs, has endo- and exonuclease activities against abasic site and other types of base damage. PALF accumulates rapidly at single-strand breaks in a poly(ADP-ribose) polymerase 1 (PARP1)-dependent manner in human cells. Indeed, PALF interacts directly with PARP1 and is required for its activation and for cellular resistance to methyl-methane sulfonate. PALF also interacts directly with KU86, LIGASEIV and phosphorylated XRCC4 proteins and possesses endo/exonuclease activity at protruding DNA ends. Various treatments that produce double-strand breaks induce formation of PALF foci, which fully coincide with γH2AX foci. Thus, PALF and the CYR motif may play important roles in DNA repair of higher eukaryotes. PMID:17396150

Gag induces the coalescence of clustered lipid rafts and tetraspanin-enriched microdomains at HIV-1 assembly sites on the plasma membrane.

Science.gov (United States)

Hogue, Ian B; Grover, Jonathan R; Soheilian, Ferri; Nagashima, Kunio; Ono, Akira

2011-10-01

The HIV-1 structural protein Gag associates with two types of plasma membrane microdomains, lipid rafts and tetraspanin-enriched microdomains (TEMs), both of which have been proposed to be platforms for HIV-1 assembly. However, a variety of studies have demonstrated that lipid rafts and TEMs are distinct microdomains in the absence of HIV-1 infection. To measure the impact of Gag on microdomain behaviors, we took advantage of two assays: an antibody-mediated copatching assay and a Förster resonance energy transfer (FRET) assay that measures the clustering of microdomain markers in live cells without antibody-mediated patching. We found that lipid rafts and TEMs copatched and clustered to a greater extent in the presence of membrane-bound Gag in both assays, suggesting that Gag induces the coalescence of lipid rafts and TEMs. Substitutions in membrane binding motifs of Gag revealed that, while Gag membrane binding is necessary to induce coalescence of lipid rafts and TEMs, either acylation of Gag or binding of phosphatidylinositol-(4,5)-bisphosphate is sufficient. Finally, a Gag derivative that is defective in inducing membrane curvature appeared less able to induce lipid raft and TEM coalescence. A higher-resolution analysis of assembly sites by correlative fluorescence and scanning electron microscopy showed that coalescence of clustered lipid rafts and TEMs occurs predominantly at completed cell surface virus-like particles, whereas a transmembrane raft marker protein appeared to associate with punctate Gag fluorescence even in the absence of cell surface particles. Together, these results suggest that different membrane microdomain components are recruited in a stepwise manner during assembly.

Site and strand specificity of UVB mutagenesis in the SUP4-o gene of yeast

International Nuclear Information System (INIS)

Armstrong, J.D.; Kunz, B.A.

1990-01-01

DNA sequencing was used to characterize 208 mutations induced in the SUP4-o tRNA gene of the yeast Saccharomyces cerevisiae by UVB (285-320 nm) radiation. The results were compared to those for an analysis of 211 SUP4-o mutations induced by 254-nm UVC light. In each case, greater than 90% of the mutations were single base-pair changes but G.C----A.T transitions predominated and accounted for more of the mutations induced by UVB than UVC. Double substitutions, single base-pair deletions, and more complex events were also recovered. However, UVB induced 3-fold more tandem substitutions than UVC and nontandem double events were detected only after irradiation with UVC. Virtually all induced substitutions occurred at sites where the pyrimidine of the base pair was part of a dipyrimidine sequence. Although the site specificities were consistent with roles for cyclobutane dimers and pyrimidine-pyrimidone(6-4) lesions in mutation induction, preliminary photoreactivation data implicated cyclobutane dimers as the major form of premutational DNA damage for both agents. Intriguingly, there was a preference for both UVB- and UVC-induced mutations to occur at sites where the dipyrimidine was on the transcribed strand

Final work plan: Expedited Site Characterization of the IES Industries, Inc., Site at Marshalltown, Iowa. Ames Expedited Site Characterization Project, Version 1.0

Energy Technology Data Exchange (ETDEWEB)

1994-04-04

The overall goal of the Ames Laboratory Expedited Site Characterization (ESC) project is to evaluate and promote both innovative and state-of-the-practice site characterization and/or monitoring technologies. This will be accomplished by fielding both types of technologies together in the context of an expedited site characterization. The first site will be at a former manufactured gas plant (FMGP) in Marshalltown, Iowa. The project will field three areas of technology: geophysical, analytical, and data fusion. Geophysical technologies are designed to understand the subsurface geology to help predict fate and transport of the target contaminants. Analytical technologies/methods are designed to detect and quantify the target contaminants. Data fusion technology consists of software systems designed to rapidly integrate or fuse all site information into a conceptual site model that then becomes the decision making tool for the site team to plan subsequent sampling activity. Not all of the contaminants present can be located at the action level. Polynuclear aromatic hydrocarbons (PAHs) are the signature organics associated with the coal tar activities that took place at the site. As a result, PAHs were selected as the target compounds. Screening analytical instruments and nonintrusive geophysical techniques will be fielded to qualitatively map the spatial contaminant distribution. Soil gas surveys, immunoassay testing (IMA), innovative optical techniques, and passive organic sorbent sensors will be deployed along with the geophysical methods. Gas chromatography/mass spectrometry (GC/MS) instruments and a cone penetrometer system equipped with a laser-induced fluorescence (LIF) probe will quantitatively map the action level edges of the PAH plume(s). Samples will be taken both by the cone penetrometer test system (CPT) and the Geoprobe {reg_sign} sampler system.

S1-sensitive sites in DNA after γ-irradiation

International Nuclear Information System (INIS)

Martin-Bertram, H.

1981-01-01

DNA from γ-irradiated T 1 bacteriophages was analyzed for 'single-stranded' sites by cleavage with S1 nuclease from Aspergillus oryzae as lesion probe. The ratio of 'S1-sensitive sites' to the amount of radiation-induced single-strand breaks was about one. Presumably these 'denatured' sites were associated with single-strand breaks. The subsequent check for the persistence of 'single-stranded' sites within the DNA molecule by thermokinetics demonstrated a strong affinity of the nuclease to its substrate, the single-stranded lesion, and a perfect excision. It is assumed that the direct absorption of radiation energy in the DNA gives rise to the formation of such bulky lesions. (Auth.)

The Influence of Heritage Sites as Filming Locations on Tourists’ Decisions to Visit Sites and Their Perceptions of Them. Case Study: Game of Thrones

Directory of Open Access Journals (Sweden)

Emily Bowyer

2017-10-01

Full Text Available This paper provides an overview of the authors’ master thesis and addresses the effects of World Heritage Sites and heritage sites which are used as filming locations on visitor perceptions of a site and their decision to visit a site. Film-induced tourism is becoming increasingly popular and it is important to assess its impacts on World Heritage Sites and heritage sites used as locations. The integration of the different aspects of heritage and filming at a site including elements and the communication between all the different parties involved are also addressed. The case study used is the popular television series Game of Thrones focusing on various locations in Northern Ireland and Dubrovnik, Croatia. The paper aims to provide a starting platform for future research on heritage sites used as filming locations and the possible impacts that this may have.

NPP Siting in Western Part of Java Island Indonesia: Regional Analysis Stage

International Nuclear Information System (INIS)

Sastratenaya, A.S.; Yuliastuti

2011-01-01

Full text of publication follows: Considering that Banten and West Java Provinces are dense regions of industry, therefore they require a large amount of electricity. Nuclear power plant is one option to be considered to anticipate the future electricity demand. To support the program, it is needed to look for some potential locations through NPP siting. The siting should meet the requirement of safety, safety aspects of the natural external events, human induced external events, public and environmental safety. Site selection is performed in several stages, where each stage has specific assessment criteria. Siting is commenced with pre-survey activity to obtain several interest areas, the activity covers a wide area but the used data is very limited and only apply general criteria. The following activities after pre survey are site survey consisting of (1) regional analysis, (2) site screening, and (3) comparison and ranking stages. The objective of regional analysis is to obtain potential sites in the study area of 150 km radius from each interest area by using both general and specific criteria. The potential sites then screened to obtain selected candidate sites by using more detailed secondary data as well as survey activities such as geophysical investigation, a few of drilling, etc., within the radius of 50 km from each potential site. All the selected candidate sites are then compared and ranked to obtain preferred candidate site. Site evaluation is the next step to evaluate all site-specific parameter to obtain design basis parameters and as the basis for preparing site permit document. This paper presents the methodology and result of regional analysis stage. The objective of the activity is to obtain potential sites in the north coast of West Java and Banten Provinces by considering fourteen study aspects which could be categorize into safety related aspects, non-safety related aspect and public education. However, this paper only considers the safety

Selectivity of externally facing ion-binding sites in the Na/K pump to alkali metals and organic cations.

Science.gov (United States)

Ratheal, Ian M; Virgin, Gail K; Yu, Haibo; Roux, Benoît; Gatto, Craig; Artigas, Pablo

2010-10-26

The Na/K pump is a P-type ATPase that exchanges three intracellular Na(+) ions for two extracellular K(+) ions through the plasmalemma of nearly all animal cells. The mechanisms involved in cation selection by the pump's ion-binding sites (site I and site II bind either Na(+) or K(+); site III binds only Na(+)) are poorly understood. We studied cation selectivity by outward-facing sites (high K(+) affinity) of Na/K pumps expressed in Xenopus oocytes, under voltage clamp. Guanidinium(+), methylguanidinium(+), and aminoguanidinium(+) produced two phenomena possibly reflecting actions at site III: (i) voltage-dependent inhibition (VDI) of outwardly directed pump current at saturating K(+), and (ii) induction of pump-mediated, guanidinium-derivative-carried inward current at negative potentials without Na(+) and K(+). In contrast, formamidinium(+) and acetamidinium(+) induced K(+)-like outward currents. Measurement of ouabain-sensitive ATPase activity and radiolabeled cation uptake confirmed that these cations are external K(+) congeners. Molecular dynamics simulations indicate that bound organic cations induce minor distortion of the binding sites. Among tested metals, only Li(+) induced Na(+)-like VDI, whereas all metals tested except Na(+) induced K(+)-like outward currents. Pump-mediated K(+)-like organic cation transport challenges the concept of rigid structural models in which ion specificity at site I and site II arises from a precise and unique arrangement of coordinating ligands. Furthermore, actions by guanidinium(+) derivatives suggest that Na(+) binds to site III in a hydrated form and that the inward current observed without external Na(+) and K(+) represents cation transport when normal occlusion at sites I and II is impaired. These results provide insights on external ion selectivity at the three binding sites.

On-Site Radon Detection of Mining-induced Fractures from Overlying Strata to the Surface: A Case Study of the Baoshan Coal Mine in China

Directory of Open Access Journals (Sweden)

Wei Zhang

2014-12-01

Full Text Available Large-scale longwall mining of shallow coal seams may cause mining-induced fractures that can project completely through to the surface. This could lead to a series of mine safety and environmental issues, further deteriorating the already fragile ecological environment in the Western mining areas in China. Therefore, an accurate and effective understanding of the spatiotemporal evolution law of mining-induced fractures in overlying strata and its relationship to upper aquifers is critical. In this paper, the application of the geophysical-chemical properties of radon in mining engineering is explored as a potential solution to the shortcomings of existing surveying methods. A radioactive measurement method is proposed for the detection of the development of mining-induced fractures from overlying strata to the surface in the Baoshan Coal Mine (BCM. The on-site test indicated that the first weighting step is approximately 60 m, the average periodic weighting step is approximately 20 m, and the influence coverage of the advanced abutment pressure is approximately 30 m. The presented method could be used as an indirect technical support to increase the safety of coal mining by acting as a simple, fast, and reliable method of detecting mining-induced fractures in overlying strata.

The impact of locally multiply damaged sites (LMDS) induced by ionizing radiation in mammalian cells

International Nuclear Information System (INIS)

Averbeck, D.; Boucher, D.

2006-01-01

Monte Carlo calculations have shown that ionising radiations produce a specific type of clustered cell damage called locally multiply damaged sites or LMDS. These lesions consist of closely positioned single-strand breaks, (oxidative) base damage and DNA double-strand breaks (DSB) in between one helical turn of DNA. As specific markers of radiation-induced damage these lesions are likely to condition biological responses and are thus of great interest for radiation protection. Calculations indicate that there should be more LMDS induced by high than by low LET radiation, and they should be absent in un-irradiated cells. Processes like K-shell activation and local Auger electron emission can be expected to add complex DSB or LMDS, producing significant chromosomal damage. In the discussion of the specificity of ionising radiation in comparison to other genotoxic agents, many arguments have been put forward that these lesions should be particularly deleterious for living cells. Complex lesions of that type should represent big obstacles for DNA repair and give rise to high lethality. Moreover, cellular attempts to repair them could accentuate harm, leading to mutations, genetic instability and cancer. In vitro experiments with oligonucleotides containing an artificially introduced set of base damage and SSB in different combinations have shown that depending on the close positioning of the damage on DNA, repair enzymes, and even whole cell extracts, are unable to repair properly and may stimulate mis-repair. Pulsed field gel electrophoresis (PFGE) in conjunction with enzymatic treatments has been used to detect LMDS in mammalian cells after high and low LET radiation. In order to further define the importance of LMDS for radiation induced cellular responses, we studied the induction of LMDS as a function of radiation dose and dose rate in mammalian cells (CHO and MRC5) using 137 Cs gamma-radiation. Using PFGE and specific glycosylases to convert oxidative damage into

The impact of locally multiply damaged sites (LMDS) induced by ionizing radiation in mammalian cells

Energy Technology Data Exchange (ETDEWEB)

Averbeck, D.; Boucher, D. [Institut Curie-Section de Recherche, UMR2027 CNRS, LCR-V28 du CEA, Centre Universitaire, 91405 Orsay Cedex (France)

2006-07-01

Monte Carlo calculations have shown that ionising radiations produce a specific type of clustered cell damage called locally multiply damaged sites or LMDS. These lesions consist of closely positioned single-strand breaks, (oxidative) base damage and DNA double-strand breaks (DSB) in between one helical turn of DNA. As specific markers of radiation-induced damage these lesions are likely to condition biological responses and are thus of great interest for radiation protection. Calculations indicate that there should be more LMDS induced by high than by low LET radiation, and they should be absent in un-irradiated cells. Processes like K-shell activation and local Auger electron emission can be expected to add complex DSB or LMDS, producing significant chromosomal damage. In the discussion of the specificity of ionising radiation in comparison to other genotoxic agents, many arguments have been put forward that these lesions should be particularly deleterious for living cells. Complex lesions of that type should represent big obstacles for DNA repair and give rise to high lethality. Moreover, cellular attempts to repair them could accentuate harm, leading to mutations, genetic instability and cancer. In vitro experiments with oligonucleotides containing an artificially introduced set of base damage and SSB in different combinations have shown that depending on the close positioning of the damage on DNA, repair enzymes, and even whole cell extracts, are unable to repair properly and may stimulate mis-repair. Pulsed field gel electrophoresis (PFGE) in conjunction with enzymatic treatments has been used to detect LMDS in mammalian cells after high and low LET radiation. In order to further define the importance of LMDS for radiation induced cellular responses, we studied the induction of LMDS as a function of radiation dose and dose rate in mammalian cells (CHO and MRC5) using {sup 137}Cs gamma-radiation. Using PFGE and specific glycosylases to convert oxidative damage

Environmental radioactivity survey in site studies

International Nuclear Information System (INIS)

Segovia, N.; Gaso, M.I.; Tejera, A.; Tamez, E.

1989-01-01

Systematic monitoring of Rn in soil, background gamma levels and meteorological measurements was conducted at the site of a nuclear research center. The data cover a time period of several years. The radon in soil gas fluctuations is influenced by seasonal meteorological changes and by local geophysical parameters and they range between 1000 Bq/m 3 and 18000 Bq/m 3 . The observed gamma levels show geographic stability; some seasonal fluctuations were observed. Data analysis indicates that no significant changes occured at the site over the measuring period and radiation levels and meteorological changes are moderate inducing no ecological impact on the surroundings. (author) 8 refs.; 5 figs.; 2 tabs

Bacterial natural transformation by highly fragmented and damaged DNA

DEFF Research Database (Denmark)

Overballe-Petersen, Søren; Harms, Klaus; Orlando, Ludovic Antoine Alexandre

2013-01-01

for microbes, but not as potential substrate for bacterial evolution. Here, we show that fragmented DNA molecules (≥20 bp) that additionally may contain abasic sites, cross-links, or miscoding lesions are acquired by the environmental bacterium Acinetobacter baylyi through natural transformation. With uptake......DNA molecules are continuously released through decomposition of organic matter and are ubiquitous in most environments. Such DNA becomes fragmented and damaged (often DNA is recognized as nutrient source...... of DNA from a 43,000-y-old woolly mammoth bone, we further demonstrate that such natural transformation events include ancient DNA molecules. We find that the DNA recombination is RecA recombinase independent and is directly linked to DNA replication. We show that the adjacent nucleotide variations...

Site-Specific Phosphorylation of Ikaros Induced by Low-Dose Ionizing Radiation Regulates Cell Cycle Progression of B Lymphoblast Through CK2 and AKT Activation

Energy Technology Data Exchange (ETDEWEB)

Cho, Seong-Jun; Kang, Hana [KHNP Radiation Health Institute, Korea Hydro & Nuclear Power Co, Seoul (Korea, Republic of); Kim, Min Young [Department of Molecular Biology, College of Natural Sciences, Pusan National University, Busan (Korea, Republic of); Lee, Jung Eun; Kim, Sung Jin; Nam, Seon Young; Kim, Ji Young; Kim, Hee Sun [KHNP Radiation Health Institute, Korea Hydro & Nuclear Power Co, Seoul (Korea, Republic of); Pyo, Suhkneung [College of Pharmacy, Sungkyunkwan University, Suwon, Gyeonggi-do (Korea, Republic of); Yang, Kwang Hee, E-mail: kwangheey@khnp.co.kr [KHNP Radiation Health Institute, Korea Hydro & Nuclear Power Co, Seoul (Korea, Republic of)

2016-04-01

Purpose: To determine how low-dose ionizing radiation (LDIR) regulates B lympho-proliferation and its molecular mechanism related with Ikaros, transcription factor. Methods and Materials: Splenocytes and IM-9 cells were uniformly irradiated with various doses of a {sup 137}Cs γ-source, and cell proliferation was analyzed. To determine the LDIR-specific phosphorylation of Ikaros, immunoprecipitation and Western blot analysis were performed. To investigate the physiologic function of LDIR-mediatied Ikaros phosphorylation, Ikaros mutants at phosphorylation sites were generated, and cell cycle analysis was performed. Results: First, we found that LDIR enhances B lymphoblast proliferation in an Ikaros-dependent manner. Moreover, we found that LDIR elevates the phosphorylation level of Ikaros protein. Interestingly, we showed that CK2 and AKT are involved in LDIR-induced Ikaros phosphorylation and capable of regulating DNA binding activity of Ikaros via specific phosphorylation. Finally, we identified LDIR-specific Ikaros phosphorylation sites at S391/S393 and showed that the Ikaros phosphorylations at these sites control Ikaros's ability to regulate G1/S cell cycle progression. Conclusion: Low-dose ionizing radiation specifically phosphorylates Ikaros protein at Ser 391/393 residues to regulate cell cycle progression in B lymphoblast.

Site-Specific Phosphorylation of Ikaros Induced by Low-Dose Ionizing Radiation Regulates Cell Cycle Progression of B Lymphoblast Through CK2 and AKT Activation.

Science.gov (United States)

Cho, Seong-Jun; Kang, Hana; Kim, Min Young; Lee, Jung Eun; Kim, Sung Jin; Nam, Seon Young; Kim, Ji Young; Kim, Hee Sun; Pyo, Suhkneung; Yang, Kwang Hee

2016-04-01

To determine how low-dose ionizing radiation (LDIR) regulates B lympho-proliferation and its molecular mechanism related with Ikaros, transcription factor. Splenocytes and IM-9 cells were uniformly irradiated with various doses of a (137)Cs γ-source, and cell proliferation was analyzed. To determine the LDIR-specific phosphorylation of Ikaros, immunoprecipitation and Western blot analysis were performed. To investigate the physiologic function of LDIR-mediatied Ikaros phosphorylation, Ikaros mutants at phosphorylation sites were generated, and cell cycle analysis was performed. First, we found that LDIR enhances B lymphoblast proliferation in an Ikaros-dependent manner. Moreover, we found that LDIR elevates the phosphorylation level of Ikaros protein. Interestingly, we showed that CK2 and AKT are involved in LDIR-induced Ikaros phosphorylation and capable of regulating DNA binding activity of Ikaros via specific phosphorylation. Finally, we identified LDIR-specific Ikaros phosphorylation sites at S391/S393 and showed that the Ikaros phosphorylations at these sites control Ikaros's ability to regulate G1/S cell cycle progression. Low-dose ionizing radiation specifically phosphorylates Ikaros protein at Ser 391/393 residues to regulate cell cycle progression in B lymphoblast. Copyright © 2016 Elsevier Inc. All rights reserved.

Site-Specific Phosphorylation of Ikaros Induced by Low-Dose Ionizing Radiation Regulates Cell Cycle Progression of B Lymphoblast Through CK2 and AKT Activation

International Nuclear Information System (INIS)

Cho, Seong-Jun; Kang, Hana; Kim, Min Young; Lee, Jung Eun; Kim, Sung Jin; Nam, Seon Young; Kim, Ji Young; Kim, Hee Sun; Pyo, Suhkneung; Yang, Kwang Hee

2016-01-01

Purpose: To determine how low-dose ionizing radiation (LDIR) regulates B lympho-proliferation and its molecular mechanism related with Ikaros, transcription factor. Methods and Materials: Splenocytes and IM-9 cells were uniformly irradiated with various doses of a "1"3"7Cs γ-source, and cell proliferation was analyzed. To determine the LDIR-specific phosphorylation of Ikaros, immunoprecipitation and Western blot analysis were performed. To investigate the physiologic function of LDIR-mediatied Ikaros phosphorylation, Ikaros mutants at phosphorylation sites were generated, and cell cycle analysis was performed. Results: First, we found that LDIR enhances B lymphoblast proliferation in an Ikaros-dependent manner. Moreover, we found that LDIR elevates the phosphorylation level of Ikaros protein. Interestingly, we showed that CK2 and AKT are involved in LDIR-induced Ikaros phosphorylation and capable of regulating DNA binding activity of Ikaros via specific phosphorylation. Finally, we identified LDIR-specific Ikaros phosphorylation sites at S391/S393 and showed that the Ikaros phosphorylations at these sites control Ikaros's ability to regulate G1/S cell cycle progression. Conclusion: Low-dose ionizing radiation specifically phosphorylates Ikaros protein at Ser 391/393 residues to regulate cell cycle progression in B lymphoblast.

Production site of radiation-induced pulmonary fibrosis

International Nuclear Information System (INIS)

Song Liangwen; Cui Xuemei; Gao Yabing; Yang Ruibiao; Xia Guowei; Wang Dewen

1997-01-01

Production site development and alterations of early pulmonary fibrosis were studied. Single irradiation was made at right thorax of rats with 0, 15 and 30 Gy of γ-irradiation, respectively. The rats were divided into three groups which were sacrificed 1, 3, 5 months post irradiation. Hydroxyproline in lungs was measured by biochemical method. Pulmonary type I and III collagens were measured by polarization method. Distribution of angiotensin II (A II) in pulmonary tissues was displayed by immunohistochemical method. Extent of pulmonary fibrosis relatively increased with irradiation dose and time elapse after irradiation. Ratio of type I to type III collagens increased with increasing fibrosis. Proliferating collagen fibers mainly came from fibroblasts of pulmonary bronchial and arterial adventitia, and extended into pulmonary parenchyma. Meanwhile, type I collagen substituted for type III collagen in interstitium of pulmonary alveoli. A II was positive for fibroblasts and macrophages in pulmonary interstitium. Irradiation can stimulate fibroblasts in interstitium proliferation, and type I collagen substitutes for type III collagen. Expression and synthesis of A II in interstitium may promote the course of pulmonary fibrosis

Gag Induces the Coalescence of Clustered Lipid Rafts and Tetraspanin-Enriched Microdomains at HIV-1 Assembly Sites on the Plasma Membrane ▿

Science.gov (United States)

Hogue, Ian B.; Grover, Jonathan R.; Soheilian, Ferri; Nagashima, Kunio; Ono, Akira

2011-01-01

The HIV-1 structural protein Gag associates with two types of plasma membrane microdomains, lipid rafts and tetraspanin-enriched microdomains (TEMs), both of which have been proposed to be platforms for HIV-1 assembly. However, a variety of studies have demonstrated that lipid rafts and TEMs are distinct microdomains in the absence of HIV-1 infection. To measure the impact of Gag on microdomain behaviors, we took advantage of two assays: an antibody-mediated copatching assay and a Förster resonance energy transfer (FRET) assay that measures the clustering of microdomain markers in live cells without antibody-mediated patching. We found that lipid rafts and TEMs copatched and clustered to a greater extent in the presence of membrane-bound Gag in both assays, suggesting that Gag induces the coalescence of lipid rafts and TEMs. Substitutions in membrane binding motifs of Gag revealed that, while Gag membrane binding is necessary to induce coalescence of lipid rafts and TEMs, either acylation of Gag or binding of phosphatidylinositol-(4,5)-bisphosphate is sufficient. Finally, a Gag derivative that is defective in inducing membrane curvature appeared less able to induce lipid raft and TEM coalescence. A higher-resolution analysis of assembly sites by correlative fluorescence and scanning electron microscopy showed that coalescence of clustered lipid rafts and TEMs occurs predominately at completed cell surface virus-like particles, whereas a transmembrane raft marker protein appeared to associate with punctate Gag fluorescence even in the absence of cell surface particles. Together, these results suggest that different membrane microdomain components are recruited in a stepwise manner during assembly. PMID:21813604

Hydrogen solution in tetrahedral or octahedral interstitial sites in Al

International Nuclear Information System (INIS)

Zeng, C.A.; Hu, J.P.; Ouyang, C.Y.

2011-01-01

Highlights: → The physical nature of the site preference for H solution in BCC Al is revealed. → The site preference is result of competition between Al-H bonding interaction and local lattice distortion. → The Al-H bonding interaction lowers the solution energy while the local lattice distortion increases the solution energy. - Abstract: It is reported that H atoms prefer to stay at interstitial (defect) sites with larger space in most metals. However, H atom prefers to occupy tetrahedral interstitial sites (T-site) that provide smaller space than octahedral sites (O-site) in Al. This paper studied the H-Al interactions from first principles calculations. Through analysis of the H-induced electronic states and the local atomic relaxations, we show that H-Al bonding interaction is stronger for T-site H, which is in favor of the solution energy. On the other hand, larger local atomic distortion is observed around the T-site H, which increases the total energy.

Tension-induced binding of semiflexible biopolymers

Science.gov (United States)

Benetatos, Panayotis; von der Heydt, Alice; Zippelius, Annette

2015-03-01

We investigate theoretically the effect of polymer tension on the collective behaviour of reversible cross-links. We use a model of two parallel-aligned, weakly-bending wormlike chains with a regularly spaced sequence of binding sites subjected to a tensile force. Reversible cross-links attach and detach at the binding sites with an affinity controlled by a chemical potential. In a mean-field approach, we calculate the free energy of the system and we show the emergence of a free energy barrier which controls the reversible (un)binding. The tension affects the conformational entropy of the chains which competes with the binding energy of the cross-links. This competition gives rise to a sudden increase in the fraction of bound sites as the polymer tension increases. The force-induced first-order transition in the number of cross-links implies a sudden force-induced stiffening of the effective stretching modulus of the polymers. This mechanism may be relevant to the formation and stress-induced strengthening of stress fibers in the cytoskeleton. We acknowledge support by the Deutsche Forschungsgemeinschaft (DFG) via grant SFB-937/A1.

Fusion proteins of HIV-1 envelope glycoprotein gp120 with CD4-induced antibodies showed enhanced binding to CD4 and CD4 binding site antibodies

Energy Technology Data Exchange (ETDEWEB)

Chen, Weizao, E-mail: chenw3@mail.nih.gov [Protein Interactions Group, Frederick National Laboratory for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702 (United States); Feng, Yang [Protein Interactions Group, Frederick National Laboratory for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702 (United States); Wang, Yanping [Protein Interactions Group, Frederick National Laboratory for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702 (United States); The Basic Research Program, Science Applications International Corporation-Frederick, Inc., National Cancer Institute, National Institutes of Health, Frederick, MD 21702 (United States); Zhu, Zhongyu; Dimitrov, Dimiter S. [Protein Interactions Group, Frederick National Laboratory for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702 (United States)

2012-09-07

Highlights: Black-Right-Pointing-Pointer Some recombinant HIV-1 gp120s do not preserve their conformations on gp140s. Black-Right-Pointing-Pointer We hypothesize that CD4i antibodies could induce conformational changes in gp120. Black-Right-Pointing-Pointer CD4i antibodies enhance binding of CD4 and CD4bs antibodies to gp120. Black-Right-Pointing-Pointer CD4i antibody-gp120 fusion proteins could have potential as vaccine immunogens. -- Abstract: Development of successful AIDS vaccine immunogens continues to be a major challenge. One of the mechanisms by which HIV-1 evades antibody-mediated neutralizing responses is the remarkable conformational flexibility of its envelope glycoprotein (Env) gp120. Some recombinant gp120s do not preserve their conformations on gp140s and functional viral spikes, and exhibit decreased recognition by CD4 and neutralizing antibodies. CD4 binding induces conformational changes in gp120 leading to exposure of the coreceptor-binding site (CoRbs). In this study, we test our hypothesis that CD4-induced (CD4i) antibodies, which target the CoRbs, could also induce conformational changes in gp120 leading to better exposed conserved neutralizing antibody epitopes including the CD4-binding site (CD4bs). We found that a mixture of CD4i antibodies with gp120 only weakly enhanced CD4 binding. However, such interactions in single-chain fusion proteins resulted in gp120 conformations which bound to CD4 and CD4bs antibodies better than the original or mutagenically stabilized gp120s. Moreover, the two molecules in the fusion proteins synergized with each other in neutralizing HIV-1. Therefore, fusion proteins of gp120 with CD4i antibodies could have potential as components of HIV-1 vaccines and inhibitors of HIV-1 entry, and could be used as reagents to explore the conformational flexibility of gp120 and mechanisms of entry and immune evasion.

Model synthetic complexes of the hydrogenase with different protonation sites; Complexes synthetiques modeles de l'hydrogenase avec differents sites de protonation

Energy Technology Data Exchange (ETDEWEB)

Capon, J.F.; Gloaguen, F.; Morvan, D.; Schollhammer, Ph.; Talarmin, J.; Yaouanc, J.J. [Universite de Bretagne Occidentale, UMR CNRS 6521, Chimie, Electrochimie Moleculaires et Chimie Analytique, Faculte des Sciences, 29 - Brest (France)

2005-07-01

The data obtained until now seem to indicate that the hydrogen production by hydrogenases induces a proton-hydride coupling. In taking the structures of theses enzymes active sites (determined by X-ray diffraction) as a basis, it can be thought that this proton-hydride coupling is facilitated by the juxtaposition of two protonation sites, the metallic center M and the basic group of an E ligand of the coordination sphere. Contrarily to the supposed running of the hydrogenases enzymes, the homogeneous catalysts of the protons reduction, described in the literature, present a reactivity which is either on an alone metallic site or on a metal-metal bond. This work deals then with the preparation of complexes having two juxtaposed protonation sites. Some iron dinuclear compounds have been synthesized and their properties studied. (O.M.)

Effect of flood-induced chemical load on filtrate quality at bank filtration sites

Science.gov (United States)

Ray, C.; Soong, T.W.; Lian, Y.Q.; Roadcap, G.S.

2002-01-01

Riparian municipal wells, that are located on riverbanks, are specifically designed to capture a portion of the river water through induced infiltration. Runoff from agricultural watersheds is found to carry enormous amounts of pesticides and nitrate. While the risk of contamination for a vast majority of sites with small-capacity vertical wells is low, potential exists for medium to large capacity collector wells to capture a fraction of the surface water contaminants during flood. Prior monitoring and current modeling results indicate that a small-capacity (peak pumpage 0.0315 m3/s) vertical bank filtration well may not be affected by river water nitrate and atrazine even during flood periods. For a medium capacity (0.0875-0.175 m3/s) hypothetical collector well at the same site, potential exists for a portion of the river water nitrate and atrazine to enter the well during flood periods. Various combinations of hydraulic conductivity of the riverbed or bank material were used. For nitrate, it was assumed either no denitrification occurred during the period of simulation or a half-life of 2 years. Equilibrium controlled sorption (organic carbon partition coefficient of 52 ml/g) and a half-life of between 7.5 and 15 weeks were considered for atrazine. Combinations of these parameters were used in various simulations. Peak concentrations of atrazine or nitrate in pumped water could vary from less than 1% to as high as 90% of that in the river. It was found that a combination of river stage, pumping rates, hydraulic properties of the riverbed and bank, and soil/pesticide properties could affect contaminant entry from river water to any of these wells. If the hydraulic conductivity of the bed and bank material were low, atrazine would not reach the pumping well with or without sorption and degradation. However, for moderately low permeable bank and bed materials, some atrazine from river water could enter a hypothetical collector well while pumping at 0.0875 m3/s. It

Peroxynitrite modified DNA presents better epitopes for anti-DNA autoantibodies in diabetes type 1 patients.

Science.gov (United States)

Tripathi, Prashant; Moinuddin; Dixit, Kiran; Mir, Abdul Rouf; Habib, Safia; Alam, Khursheed; Ali, Asif

2014-07-01

Peroxynitrite (ONOO(-)), formed by the reaction between nitric oxide (NO) and superoxide (O2(-)), has been implicated in the etiology of numerous disease processes. Peroxynitrite interacts with DNA via direct oxidative reactions or via indirect radical-mediated mechanism. It can inflict both oxidative and nitrosative damages on DNA bases, generating abasic sites, resulting in the single strand breaks. Plasmid pUC 18 isolated from Escherichiacoli was modified with peroxynitrite, generated by quenched flow process. Modifications incurred in plasmid DNA were characterized by ultraviolet and fluorescence spectroscopy, circular dichroism, HPLC and melting temperature studies. Binding characteristics and specificity of antibodies from diabetes patients were analyzed by direct binding and inhibition ELISA. Peroxynitrite modification of pUC 18 plasmid resulted in the formation of strand breaks and base modification. The major compound formed when peroxynitrite reacted with DNA was 8-nitroguanine, a specific marker for peroxynitrite induced DNA damage in inflamed tissues. The concentration of 8-nitroguanine was found to be 3.8 μM. Sera from diabetes type 1 patients from different age groups were studied for their binding to native and peroxynitrite modified plasmid. Direct binding and competitive-inhibition ELISA results showed higher recognition of peroxynitrite modified plasmid, as compared to the native form, by auto-antibodies present in diabetes patients. The preferential recognition of modified plasmid by diabetes autoantibodies was further reiterated by gel shift assay. Experimentally induced anti-peroxynitrite-modified plasmid IgG was used as a probe to detect nitrosative lesions in the DNA isolated from diabetes patients. Copyright © 2014 Elsevier Inc. All rights reserved.

Impairment of Neutrophil Migration to Remote Inflammatory Site during Lung Histoplasmosis

Directory of Open Access Journals (Sweden)

Alexandra I. Medeiros

2015-01-01

Full Text Available Histoplasma capsulatum (Hc induces a pulmonary disease in which leukotrienes promote activation and recruitment of effectors cells. It is also well-recognized that leukotriene B4 (LTB4 and platelet-activating factor (PAF induce leukocyte recruitment to inflammatory sites. We investigated the impact of pulmonary Hc infection on PMN migration to a remote inflammatory site. Our results show that pulmonary Hc infection impairs LTB4- or PAF-stimulated PMN recruitment to air pouch. Yet, remote inflammation did not modify PMN numbers in the bronchoalveolar lavage fluid (BALF of Hc-infected mice. Interestingly, the concomitant administration of PAF and LTB4 receptor antagonists inhibited PMN recruitment to both BALF and the remote site, demonstrating cooperation between both mediators. Along that line, our results show that PAF-elicited PMN chemotaxis was abrogated in 5-lipoxygenase-deficient animals. These results suggest caution in the indiscriminate use of anti-inflammatory drugs during infectious diseases.

Comparison of snoring sounds between natural and drug-induced sleep recorded using a smartphone.

Science.gov (United States)

Koo, Soo Kweon; Kwon, Soon Bok; Moon, Ji Seung; Lee, Sang Hoon; Lee, Ho Byung; Lee, Sang Jun

2018-08-01

Snoring is an important clinical feature of obstructive sleep apnea (OSA), and recent studies suggest that the acoustic quality of snoring sounds is markedly different in drug-induced sleep compared with natural sleep. However, considering differences in sound recording methods and analysis parameters, further studies are required. This study explored whether acoustic analysis of drug-induced sleep is useful as a screening test that reflects the characteristics of natural sleep in snoring patients. The snoring sounds of 30 male subjects (mean age=41.8years) were recorded using a smartphone during natural and induced sleep, with the site of vibration noted during drug-induced sleep endoscopy (DISE); then, we compared the sound intensity (dB), formant frequencies, and spectrograms of snoring sounds. Regarding the intensity of snoring sounds, there were minor differences within the retrolingual level obstruction group, but there was no significant difference between natural and induced sleep at either obstruction site. There was no significant difference in the F 1 and F 2 formant frequencies of snoring sounds between natural sleep and induced sleep at either obstruction site. Compared with natural sleep, induced sleep was slightly more irregular, with a stronger intensity on the spectrogram, but the spectrograms showed the same pattern at both obstruction sites. Although further studies are required, the spectrograms and formant frequencies of the snoring sounds of induced sleep did not differ significantly from those of natural sleep, and may be used as a screening test that reflects the characteristics of natural sleep according to the obstruction site. Copyright © 2017 Elsevier B.V. All rights reserved.

HIV integration sites and implications for maintenance of the reservoir.

Science.gov (United States)

Symons, Jori; Cameron, Paul U; Lewin, Sharon R

2018-03-01

To provide an overview of recent research of how HIV integration relates to productive and latent infection and implications for cure strategies. How and where HIV integrates provides new insights into how HIV persists on antiretroviral therapy (ART). Clonal expansion of infected cells with the same integration site demonstrates that T-cell proliferation is an important factor in HIV persistence, however, the driver of proliferation remains unclear. Clones with identical integration sites harbouring defective provirus can accumulate in HIV-infected individuals on ART and defective proviruses can express RNA and produce protein. HIV integration sites differ in clonally expanded and nonexpanded cells and in latently and productively infected cells and this influences basal and inducible transcription. There is a growing number of cellular proteins that can alter the pattern of integration to favour latency. Understanding these pathways may identify new interventions to eliminate latently infected cells. Using advances in analysing HIV integration sites, T-cell proliferation of latently infected cells is thought to play a major role in HIV persistence. Clonal expansion has been demonstrated with both defective and intact viruses. Production of viral RNA and protein from defective viruses may play a role in driving chronic immune activation. The site of integration may determine the likelihood of proliferation and the degree of basal and induced transcription. Finally, host factors and gene expression at the time of infection may determine the integration site. Together these new insights may lead to novel approaches to elimination of latently infected cells.

Seismic activity parameters of the Finnish potential repository sites

International Nuclear Information System (INIS)

Saari, J.

2000-10-01

Posiva Oy has started a project for estimating the possible earthquake induced rock movements on the deposition holes containing canisters of spent nuclear fuel. These estimates will be made for the four investigation sites, Romuvaara, Kivetty, Olkiluoto and Haestholmen. This study deals with the current and future seismicity associated with the above mentioned sites. Seismic belts that participate the seismic behaviour of the studied sites have been identified and the magnitude-frequency distributions of these belts have been estimated. The seismic activity parameters of the sites have been deduced from the characteristics of the seismic belts in order to forecast the seismicity during the next 100,000 years. The report discusses the possible earthquakes induced by future glaciation. The seismic interpretation seems to indicate that the previous postglacial faults in Finnish Lapland have been generated in compressional environment. The orientation of the rather uniform compression has been NW-SE, which coincide with the current stress field. It seems that, although the impact of postglacial crustal rebound must have been significant, the impact of plate tectonics has been dominant. A major assumption of this study has been that future seismicity will generally resemble the current seismicity. However, when the postglacial seismicity is concerned, the magnitude-frequency distribution is likely different and the expected maximum magnitude will be higher. Maximum magnitudes of future postglacial earthquakes have been approximated by strain release examinations. Seismicity has been examined within the framework of the lineament maps, in order to associate the future significant earthquakes with active fault zones in the vicinity of the potential repository sites. (orig.)

Soil biological attributes in arsenic-contaminated gold mining sites after revegetation.

Science.gov (United States)

Dos Santos, Jessé Valentim; de Melo Rangel, Wesley; Azarias Guimarães, Amanda; Duque Jaramillo, Paula Marcela; Rufini, Márcia; Marra, Leandro Marciano; Varón López, Maryeimy; Pereira da Silva, Michele Aparecida; Fonsêca Sousa Soares, Cláudio Roberto; de Souza Moreira, Fatima Maria

2013-12-01

Recovery of arsenic contaminated areas is a challenge society faces throughout the world. Revegetation associated with microbial activity can play an essential role in this process. This work investigated biological attributes in a gold mining area with different arsenic contents at different sites under two types of extant revegetation associated with cover layers of the soil: BS, Brachiaria sp. and Stizolobium sp., and LEGS, Acacia crassicarpa, A. holosericea, A. mangium, Sesbania virgata, Albizia lebbeck and Pseudosamanea guachapele. References were also evaluated, comprising the following three sites: B1, weathered sulfide substrate without revegetation; BM, barren material after gold extraction and PRNH (private reserve of natural heritage), an uncontaminated forest site near the mining area. The organic and microbial biomass carbon contents and substrate-induced respiration rates for these sites from highest to lowest were: PRNH > LEGS > BS > B1 and BM. These attributes were negatively correlated with soluble and total arsenic concentration in the soil. The sites that have undergone revegetation (LEGS and BS) had higher densities of bacteria, fungi, phosphate solubilizers and ammonium oxidizers than the sites without vegetation. Principal component analysis showed that the LEGS site grouped with PRNH, indicating that the use of leguminous species associated with an uncontaminated soil cover layer contributed to the improvement of the biological attributes. With the exception of acid phosphatase, all the biological attributes were indicators of soil recovery, particularly the following: microbial carbon, substrate-induced respiration, density of culturable bacteria, fungi and actinobacteria, phosphate solubilizers and metabolic quotient.

In vitro gap-directed translesion DNA synthesis of an abasic site involving human DNA polymerases epsilon, lambda, and beta

Czech Academy of Sciences Publication Activity Database

Villani, G.; Hübscher, U.; Gironis, N.; Parkkinen, S.; Pospiech, H.; Shevelev, Igor; di Cicco, G.; Markennen, E.; Syvaaja, J.E.; Le Gac, N.T.

2011-01-01

Roč. 286, č. 37 (2011), s. 32094-32104 ISSN 0021-9258 Grant - others:Academy of Finland(FI) 106986; Academy of Finland(FI) 123082 Institutional research plan: CEZ:AV0Z50520514 Keywords : DNA damage * DNA polymerase * DNA repair * DNA replication * DNA -protein interaction Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.773, year: 2011

Single-stranded γPNAs for in vivo site-specific genome editing via Watson-Crick recognition.

Science.gov (United States)

Bahal, Raman; Quijano, Elias; McNeer, Nicole A; Liu, Yanfeng; Bhunia, Dinesh C; Lopez-Giraldez, Francesco; Fields, Rachel J; Saltzman, William M; Ly, Danith H; Glazer, Peter M

2014-01-01

Triplex-forming peptide nucleic acids (PNAs) facilitate gene editing by stimulating recombination of donor DNAs within genomic DNA via site-specific formation of altered helical structures that further stimulate DNA repair. However, PNAs designed for triplex formation are sequence restricted to homopurine sites. Herein we describe a novel strategy where next generation single-stranded gamma PNAs (γPNAs) containing miniPEG substitutions at the gamma position can target genomic DNA in mouse bone marrow at mixed-sequence sites to induce targeted gene editing. In addition to enhanced binding, γPNAs confer increased solubility and improved formulation into poly(lactic-co-glycolic acid) (PLGA) nanoparticles for efficient intracellular delivery. Single-stranded γPNAs induce targeted gene editing at frequencies of 0.8% in mouse bone marrow cells treated ex vivo and 0.1% in vivo via IV injection, without detectable toxicity. These results suggest that γPNAs may provide a new tool for induced gene editing based on Watson-Crick recognition without sequence restriction.

The responsibility of business enterprises to restore access to essential public service at resettlement sites

NARCIS (Netherlands)

van der Ploeg, Lidewij; Vanclay, Francis; Lourenço, Ivo; Hesselman, Marlies; Hallo de Wolf, Antenor; Toebes, Brigit

2017-01-01

This chapter examines the provision of essential public services in resettlement sites associated with project induced displacement. Restoring and improving access to essential public services in resettlement sites is an important aspect of livelihood restoration of affected peoples. Project

Orientational ordering and site structure of SiF4 trapped in rare gas solids

Science.gov (United States)

Jones, Llewellyn H.; Swanson, Basil I.; Ekberg, Scott A.

1984-12-01

The infrared spectrum of the ν3 mode of SiF4 shows only one site with marked site-symmetry splitting accompanied by pronounced (>95%) orientational ordering. The results indicate a site symmetry of C3v with the SiF4 molecule occupying a four-atom substitutional site with a cubic-close-packed environment. In krypton and xenon matrices, however, SiF4 occupies a three-atom substitutional site. The fact that SiF4 shows only one dominant site when present in argon matrices of UF6 indicates that one of the two UF6 sites has a local hexagonal close packed environment induced by short range stacking faults.

Chemical-induced Vitiligo

Science.gov (United States)

Harris, John E.

2016-01-01

Synopsis Chemical-induced depigmentation of the skin has been recognized for over 75 years, first as an occupational hazard but then extending to those using household commercial products as common as hair dyes. Since their discovery, these chemicals have been used therapeutically in patients with severe vitiligo to depigment their remaining skin and improve their appearance. The importance of recognizing this phenomenon was highlighted during an outbreak of vitiligo in Japan during the summer of 2013, when over 16,000 users of a new skin lightening cosmetic cream developed skin depigmentation at the site of contact with the cream and many in remote areas as well. Depigmenting chemicals appear to be analogs of the amino acid tyrosine that disrupt melanogenesis and result in autoimmunity and melanocyte destruction. Because chemical-induced depigmentation is clinically and histologically indistinguishable from non-chemically induced vitiligo, and because these chemicals appear to induce melanocyte autoimmunity, this phenomenon should be known as “chemical-induced vitiligo”, rather than less accurate terms that have been previously used. PMID:28317525

SiteChar. Characterisation of European CO2 storage. Deliverable 8.4. Quantitative social site characterisations

Energy Technology Data Exchange (ETDEWEB)

Brunsting, S.; Mastop, E.A. [ECN Policy Studies, Energy research Centre of the Netherlands ECN, Amsterdam (Netherlands); Kaiser, M.; Zimmer, R. [Unabhaengiges Institut fuer Umweltfragen UfU, Berlin (Germany)

2013-06-15

This report describes the results of the last stage of the in-depth social site characterisation activities at two prospective CCS sites as part of the SiteChar project: a CCS onshore site and a CCS offshore site. The onshore site is the Zalecze and Zuchlow site application (Poland - WP5) and the offshore site is the North Sea Moray Firth site (UK - WP3). This deliverable describes the results from a repeated quantitative measurement of local awareness, knowledge, and perceptions of CCS at both sites using representative surveys. For comparison and discussion of all SiteChar WP8 results we refer to the final summary report D8.5. The 2nd survey showed some interesting results. First of all, awareness of CCS was still very low. While in the UK around half of the respondents had at least heard of local plans for CCS, in Poland this was only 21%. It seems that awareness in the UK was mostly induced by specific plans in the area that were abandoned in the course of the SiteChar project. Second, it seems that on the whole the local publics were rather positive about CCS. Most respondents expected a positive impact of CCS on the region. In the UK, arguments for that were mainly economic, while in Poland arguments were mainly related to environmental concerns. Although there are some worries about risks of leakage, especially at the onshore site in Poland, people think that authorities will properly regulate CCS and monitor the safety of CCS. Expectations were mostly that it would be good for the country and that it will help reach international targets for CO2 reduction and buy time to develop renewable energy. Respondents seemed uncertain about the costs of using CCS and whether the technique is ready for widespread use. Especially in Poland people seemed to agree that CCS is essential for tackling climate change. Most differences between the two sites may be attributed to the proximity of the site to the local community. The Polish site is onshore and therefore much

Repair of radiation-induced heat-labile sites is independent of DNA-PKcs, XRCC1 or PARP

Energy Technology Data Exchange (ETDEWEB)

Stenerl& #246; w, Bo; Karlsson, Karin H.; Radulescu, Irina; Rydberg, Bjorn; Stenerlow, Bo

2008-04-29

Ionizing radiation induces a variety of different DNA lesions: in addition to the most critical DNA damage, the DSB, numerous base alterations, SSBs and other modifications of the DNA double-helix are formed. When several non-DSB lesions are clustered within a short distance along DNA, or close to a DSB, they may interfere with the repair of DSBs and affect the measurement of DSB induction and repair. We have previously shown that a substantial fraction of DSBs measured by pulsed-field gel electrophoresis (PFGE) are in fact due to heat-labile sites (HLS) within clustered lesions, thus reflecting an artifact of preparation of genomic DNA at elevated temperature. To further characterize the influence of HLS on DSB induction and repair, four human cell lines (GM5758, GM7166, M059K, U-1810) with apparently normal DSB rejoining were tested for bi-phasic rejoining after gamma irradiation. When heat-released DSBs were excluded from the measurements the fraction of fast rejoining decreased to less than 50% of the total. However, neither the half-times of the fast (t{sub 1/2} = 7-8 min) or slow (t{sub 1/2} = 2.5 h) DSB rejoining were changed significantly. At t=0 the heat-released DSBs accounted for almost 40% of the DSBs, corresponding to 10 extra DSB/cell/Gy in the initial DSB yield. These heat-released DSBs were repaired within 60-90 min in all tested cells, including M059K cells treated with wortmannin or DNA-PKcs defect M059J cells. Furthermore, cells lacking XRCC1 or Poly(ADP-ribose) polymerase-1 (PARP-1) rejoined both total DSBs and heat-released DSBs similar to normal cells. In summary, the presence of heat-labile sites have a substantial impact on DSB induction yields and DSB rejoining rates measured by pulsed-field gel electrophoresis, and HLS repair is independent of DNA-PKcs, XRCC1 and PARP.

Cold-induced vasoconstriction at forearm and hand skin sites: the effect of age.

Science.gov (United States)

Kingma, B R M; Frijns, A J H; Saris, W H M; van Steenhoven, A A; van Marken Lichtenbelt, W D

2010-07-01

During mild cold exposure, elderly are at risk of hypothermia. In humans, glabrous skin at the hands is well adapted as a heat exchanger. Evidence exists that elderly show equal vasoconstriction due to local cooling at the ventral forearm, yet no age effects on vasoconstriction at hand skin have been studied. Here, we tested the hypotheses that at hand sites (a) elderly show equal vasoconstriction due to local cooling and (b) elderly show reduced response to noradrenergic stimuli. Skin perfusion and mean arterial pressure were measured in 16 young adults (Y: 18-28 years) and 16 elderly (E: 68-78 years). To study the effect of local vasoconstriction mechanisms local sympathetic nerve terminals were blocked by bretylium (BR). Baseline local skin temperature was clamped at 33 degrees C. Next, local temperature was reduced to 24 degrees C. After 15 min of local cooling, noradrenaline (NA) was administered to study the effect of neural vasoconstriction mechanisms. No significant age effect was observed in vasoconstriction due to local cooling at BR sites. After NA, vasoconstriction at the forearm showed a significant age effect; however, no significant age effect was found at the hand sites. [Change in CVC (% from baseline): Forearm Y: -76 +/- 3 vs. E: -60 +/- 5 (P forearm, elderly did not show a blunted response to local cooling and noradrenaline at hand skin sites. This indicates that at hand skin the noradrenergic mechanism of vasoconstriction is maintained with age.

A novel topical protectant for the prevention of β-radiation induced moist desquamation

International Nuclear Information System (INIS)

Ma, L.; Wilcock, S.; Rezvani, M.; Hsia, C.

2003-01-01

Full text: Effective therapies for the prevention of radiation-induced skin burns that could be readily deployed under a nuclear accident or nuclear terrorism scenario are urgently needed. In this report we describe the efficacy of a novel radioprotectant (DMZ911) in a model of b-radiation induced moist desquamation (MD) in pig skin. DMZ911 is a nitroxide-based topical cream that effectively delivers the nitroxide into viable skin cells. Stable nitroxide compounds have been shown to be effective against both X-ray and ?-ray-induced damage in vivo and in vitro. A pig skin model of β-radiation-induced MD was employed in this study. Exposure to 30 Gy was used to induce skin lesions involving >80% moist desquamation in prescribed test sites on flank skin of female Large White pigs. DMZ911 or placebo was applied to various test sites 2 hours prior to radiation exposure. Lesions were scored based on the area of the test site containing 50% MD (severe) as determined by clinical assessment using blinded observers. Treatment with DMZ911 resulted in a 31% net reduction in MD when compared to placebo treated sites following an 8-week study period. This reduction was observed whether all sites or only those with severe MD were considered. Skin damage (as indicated by MD) from radiation exposure was significantly reduced by 31% (p = 0.05) following pretreatment with the novel topical radioprotectant DMZ911. This observation suggests that skin lesion development from radiation-induced oxidative damage cascades may be successfully inhibited by treatment with DMZ911. This topical therapeutic agent represents a novel treatment for nuclear radiation induced skin injury. DMZ911 may have unique applications in radiation oncology, cosmetic and therapeutic UV, laser, glycolic and dermabrasion procedures

New concept of siting in the 21st century

International Nuclear Information System (INIS)

Nakayama, Katsuma; Otsubo, Fumiharu; Kumagai, Shigeru

1995-01-01

The New Concept of Siting in the 21st Century -- the theme of this paper -- is the concept of a new siting technology which combines the application of man - made islands (offshore), underground (steep mountains along the sea), and quaternary ground with the symbiosis with local communities. It is intended not only for promotion of the construction of nuclear power plants but also for the development of a 'new community', including the inducement of enterprises to the construction site and the promotion of new industries at the site. In studying the new concept, it is necessary to select model areas, discuss design and construction methods, and make a rough calculation of construction cost. In addition, we plan to estimate economic effects of the new concept on the construction site area. During 1993, we discussed appropriate physical conditions (topographic feature, geological structure, etc.) and geographical conditions (population density, national parks, fishing right, etc.) for the application of the siting technology giving consideration to the concept symbiosis with local communities. At present, the estimations of construction cost and future effects of new siting technology on local communities are carried on to study the new concept of siting in the 21st century. (author)

The oxidative potential and biological effects induced by PM10 obtained in Mexico City and at a receptor site during the MILAGRO Campaign

International Nuclear Information System (INIS)

Quintana, Raul; Serrano, Jesus; Gomez, Virginia; Foy, Benjamin de; Miranda, Javier; Garcia-Cuellar, Claudia; Vega, Elizabeth; Vazquez-Lopez, Ines; Molina, Luisa T.; Manzano-Leon, Natalia; Rosas, Irma; Osornio-Vargas, Alvaro R.

2011-01-01

As part of a field campaign that studied the impact of Mexico City pollution plume at the local, sub-regional and regional levels, we studied transport-related changes in PM 10 composition, oxidative potential and in vitro toxicological patterns (hemolysis, DNA degradation). We collected PM 10 in Mexico City (T 0 ) and at a suburban-receptor site (T 1 ), pooled according to two observed ventilation patterns (T 0 → T 1 influence and non-influence). T 0 samples contained more Cu, Zn, and carbon whereas; T 1 samples contained more of Al, Si, P, S, and K (p 4 -2 increased in T 1 during the influence periods. Oxidative potential correlated with Cu/Zn content (r = 0.74; p 1 PM 10 induced greater hemolysis and T 0 PM 10 induced greater DNA degradation. Influence/non-influence did not affect oxidative potential nor biological effects. Results indicate that ventilation patterns had little effect on intrinsic PM 10 composition and toxicological potential, which suggests a significant involvement of local sources. - Highlights: → Transport-related changes in PM 10 composition, oxidative potential and in vitro toxicity were studied. → Cu, Zn, and carbon levels were predominant in urban PM 10 ; receptor site PM 10 was rich in soil elements. → SO 4 -2 was the only component increased in PM 10 from the receptor during the influence periods. → PM 10 oxidative potential correlates with Cu/Zn content but not with studied biological effects. → Ventilation patterns had little effect on PM 10 composition and toxicity. - Mexico City ventilation patterns had little effect on the intrinsic PM 10 composition and toxicological potential, which suggests a significant involvement of local sources as opposed to downwind transport.

Curcumin-induced histone acetylation inhibition improves stress-induced gastric ulcer disease in rats.

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He, Ping; Zhou, Renmin; Hu, Guorui; Liu, Zhifeng; Jin, Yu; Yang, Guang; Li, Mei; Lin, Qian

2015-03-01

Curcumin is known to possess anti‑inflammatory properties. Despite the fact that curcumin is known to be a strong inhibitor of H+, K+‑ATPase activity, the mechanism underlying the curcumin‑induced inhibition of the transcription of the H+, K+‑ATPase α subunit in gastric mucosal parietal cells remains unclear. The present study investigated the possible mechanism by which curcumin inhibits stomach H+, K+‑ATPase activity during the acute phase of gastric ulcer disease. A rat model of stress‑induced gastric ulcers was produced, in which the anti‑ulcer effects of curcumin were examined. Curcumin‑induced inhibition of the H+, K+‑ATPase promoter via histone acetylation, was verified using a chromatin immunoprecipitation assay. The results showed that curcumin improved stress‑induced gastric ulcer disease in rats, as demonstrated by increased pH values and reduced gastric mucosal hemorrhage and ulcer index. These effects were accompanied by a significant reduction in the level of histone H3 acetylation at the site of the H+, K+‑ATPase promoter and in the expression of the gastric H+,K+‑ATPase α subunit gene and protein. In conclusion, curcumin downregulated the acetylation of histone H3 at the site of the H+, K+‑ATPase promoter gene, thereby inhibiting the transcription and expression of the H+, K+‑ATPase gene. Curcumin was shown to have a preventive and therapeutic effect in gastric ulcer disease.

An investigation of Laser Induced Breakdown Spectroscopy for use as a control in the laser removal of rock from fossils found at the Malapa hominin site, South Africa

International Nuclear Information System (INIS)

Roberts, D.E.; Plessis, A. du; Steyn, J.; Botha, L.R.; Pityana, S.; Berger, L.R.

2012-01-01

Laser Induced Breakdown Spectroscopy (LIBS) was used to study the spectra from fossils and surrounding rock recovered from the Cradle of Mankind site at Malapa, South Africa. The objective was to find a suitable spectral line(s), specific to fossils, which could be used as a control signal to limit damage to fossils during high speed laser removal of the encasing rock. The calcified clastic matrix (rock) encasing the fossils was found to emit a variety of complex LIBS spectra. Nevertheless, it was found possible to distinguish fossils in a single LIBS pulse, and without significant damage to the fossil, using spectral lines of neutral phosphorus. - Highlights: ► LIBS used to discriminate fossils from rock as potential processing control mechanism. ► 2 million year old fossils from Malapa hominin site found to be high in phosphorus. ► Rock spectral lines from silicon, iron and manganese, but no phosphorus. ► Holds great promise for process control in laser preparation of fossils. ► Also promising for accurate identification of fossils at excavation sites.

Site characterization and performance assessment for a low-level radioactive waste management site in the American Southwest

International Nuclear Information System (INIS)

Shott, G.J.; Sully, M.J.; Muller, C.J.; Hammermeister, D.P.; Ginanni, J.M.

1995-01-01

The Area 5 Radioactive Waste Management Site located in southern Nevada, has been used for the disposal of low-level radioactive waste since 1961. The site is located in the Mohave Desert of the American Southwest, an extremely arid region receiving as little as 0.1 m/yr of precipitation. Site characterization studies have measured the physical, hydrologic, and geochemical properties of core samples collected from 10 shallow boreholes and 3 deep boreholes that extend through the unsaturated zone to the uppermost aquifer. Results indicate that the unsaturated zone consists of 240 m of dry alluvial sediments and is remarkably uniform with respect to most physical parameters. Measurements of saturated hydraulic conductivity with depth showed no evidence of trends, layering, or anisotropy. Parameters for hydraulic functions were not highly variable and exhibited little trend with depth. Water potential profiles indicate that water movement in the upper alluvium is upward, except immediately following a precipitation event. Below the evaporative zone, the liquid flux was downward and of the same order of magnitude as the upward thermal vapor flux induced by the geothermal gradient. The extreme climatic conditions at the site reduce or eliminate many radionuclide release and transport mechanisms. Downward transport of radionuclides to the uppermost aquifer appears unlikely under current climatic conditions. Important radionuclide transport pathways appear to be limited to upward diffusion and advection of gases and biologically-mediated transport. Conceptual models of disposal site performance have been developed based on site characterization studies. The limited transport pathways and limited land use potential of the site provide reasonable assurance that regulatory performance objectives can be met

UVB-induced systemic immunosuppression: role of mast cells and histamine

International Nuclear Information System (INIS)

Hart, P.H.; Grimbaldeston, M.A.; Finlay-Jones, J.J.

1999-01-01

Full text: UVB radiation (290-320 nm) is immunosuppressive by multiple mechanisms allowing the outgrowth of UV-induced tumours in both mouse and man. Furthermore, patients with non-melanoma skin cancers have a higher risk of death from other cancers which could be explained by UV-induced immunomodulation. The mechanism(s) of suppression by UVB depend on whether the sensitising antigen is applied to the irradiated site ('local') or to non-irradiated sites ('systemic'). In the former, the activity of UV-induced TNFα is important as it affects the migration of Langerhans cells to draining lymph nodes. In contrast, histamine from dermal mast cells is critical to the early events by which UVB can suppress systemic immune responses. The prevalence of dermal mast cells in 7 strains and substrains of mice correlates directly with their susceptibility to UVB-induced systemic immunosuppression. Furthermore, mast cell depleted mice (Wf/Wf) are resistant to UVB-induced systemic immunomodulation. However, they become susceptible after reconstitution of the site to be irradiated with bone marrow derived mast cell precursors. The mice also gain susceptibility to cis-urocanic acid-induced systemic immunomodulation. There is considerable evidence that histamine is the mast cell product critical to downstream immunosuppressive events. Firstly, physiological concentrations of histamine suppress contact hypersensitivity responses. Secondly, histamine receptor antagonists halve UVB-induced systemic immunosuppression. Thirdly, mice with different UVB-susceptibilities are equally susceptible to histamine-induced immunosuppression, and finally, histamine can suppress contact hypersensitivity responses in Wf/Wf mice. We suggest that histamine may be immunomodulatory by multiple pathways. Histamine can induce the production of immunosuppressive prostanoids from keratinocytes. A lymphocyte-derived, histamine-induced suppressor factor was reported in the 1970's. More recently histamine has

Mitochondrial DNA maintenance is regulated in human hepatoma cells by glycogen synthase kinase 3β and p53 in response to tumor necrosis factor α.

Science.gov (United States)

Vadrot, Nathalie; Ghanem, Sarita; Braut, Françoise; Gavrilescu, Laura; Pilard, Nathalie; Mansouri, Abdellah; Moreau, Richard; Reyl-Desmars, Florence

2012-01-01

During chronic liver inflammation, up-regulated Tumor Necrosis Factor alpha (TNF-α) targets hepatocytes and induces abnormal reactive oxygen species (ROS) production responsible for mitochondrial DNA (mtDNA) alterations. The serine/threonine Glycogen Synthase Kinase 3 beta (GSK3β) plays a pivotal role during inflammation but its involvement in the maintenance of mtDNA remains unknown. The aim of this study was to investigate its involvement in TNF-α induced mtDNA depletion and its interrelationship with p53 a protein known to maintain mtDNA copy numbers. Using quantitative polymerase chain reaction (qPCR) we found that at 30 min in human hepatoma HepG2 cells TNF-α induced 0.55±0.10 mtDNA lesions per 10 Kb and a 52.4±2.8% decrease in mtDNA content dependent on TNF-R1 receptor and ROS production. Both lesions and depletion returned to baseline from 1 to 6 h after TNF-α exposure. Luminol-amplified chemiluminescence (LAC) was used to measure the rapid (10 min) and transient TNF-α induced increase in ROS production (168±15%). A transient 8-oxo-dG level of 1.4±0.3 ng/mg DNA and repair of abasic sites were also measured by ELISA assays. Translocation of p53 to mitochondria was observed by Western Blot and co-immunoprecipitations showed that TNF-α induced p53 binding to GSK3β and mitochondrial transcription factor A (TFAM). In addition, mitochondrial D-loop immunoprecipitation (mtDIP) revealed that TNF-α induced p53 binding to the regulatory D-loop region of mtDNA. The knockdown of p53 by siRNAs, inhibition by the phosphoSer(15)p53 antibody or transfection of human mutant active GSK3βS9A pcDNA3 plasmid inhibited recovery of mtDNA content while blockade of GSK3β activity by SB216763 inhibitor or knockdown by siRNAs suppressed mtDNA depletion. This study is the first to report the involvement of GSK3β in TNF-α induced mtDNA depletion. We suggest that p53 binding to GSK3β, TFAM and D-loop could induce recovery of mtDNA content through mtDNA repair.

Mitochondrial DNA maintenance is regulated in human hepatoma cells by glycogen synthase kinase 3β and p53 in response to tumor necrosis factor α.

Directory of Open Access Journals (Sweden)

Nathalie Vadrot

Full Text Available During chronic liver inflammation, up-regulated Tumor Necrosis Factor alpha (TNF-α targets hepatocytes and induces abnormal reactive oxygen species (ROS production responsible for mitochondrial DNA (mtDNA alterations. The serine/threonine Glycogen Synthase Kinase 3 beta (GSK3β plays a pivotal role during inflammation but its involvement in the maintenance of mtDNA remains unknown. The aim of this study was to investigate its involvement in TNF-α induced mtDNA depletion and its interrelationship with p53 a protein known to maintain mtDNA copy numbers. Using quantitative polymerase chain reaction (qPCR we found that at 30 min in human hepatoma HepG2 cells TNF-α induced 0.55±0.10 mtDNA lesions per 10 Kb and a 52.4±2.8% decrease in mtDNA content dependent on TNF-R1 receptor and ROS production. Both lesions and depletion returned to baseline from 1 to 6 h after TNF-α exposure. Luminol-amplified chemiluminescence (LAC was used to measure the rapid (10 min and transient TNF-α induced increase in ROS production (168±15%. A transient 8-oxo-dG level of 1.4±0.3 ng/mg DNA and repair of abasic sites were also measured by ELISA assays. Translocation of p53 to mitochondria was observed by Western Blot and co-immunoprecipitations showed that TNF-α induced p53 binding to GSK3β and mitochondrial transcription factor A (TFAM. In addition, mitochondrial D-loop immunoprecipitation (mtDIP revealed that TNF-α induced p53 binding to the regulatory D-loop region of mtDNA. The knockdown of p53 by siRNAs, inhibition by the phosphoSer(15p53 antibody or transfection of human mutant active GSK3βS9A pcDNA3 plasmid inhibited recovery of mtDNA content while blockade of GSK3β activity by SB216763 inhibitor or knockdown by siRNAs suppressed mtDNA depletion. This study is the first to report the involvement of GSK3β in TNF-α induced mtDNA depletion. We suggest that p53 binding to GSK3β, TFAM and D-loop could induce recovery of mtDNA content through mtDNA repair.

Identification of serine 348 on the apelin receptor as a novel regulatory phosphorylation site in apelin-13-induced G protein-independent biased signaling.

Science.gov (United States)

Chen, Xiaoyu; Bai, Bo; Tian, Yanjun; Du, Hui; Chen, Jing

2014-11-07

Phosphorylation plays vital roles in the regulation of G protein-coupled receptor (GPCR) functions. The apelin and apelin receptor (APJ) system is involved in the regulation of cardiovascular function and central control of body homeostasis. Here, using tandem mass spectrometry, we first identified phosphorylated serine residues in the C terminus of APJ. To determine the role of phosphorylation sites in APJ-mediated G protein-dependent and -independent signaling and function, we induced a mutation in the C-terminal serine residues and examined their effects on the interaction between APJ with G protein or GRK/β-arrestin and their downstream signaling. Mutation of serine 348 led to an elimination of both GRK and β-arrestin recruitment to APJ induced by apelin-13. Moreover, APJ internalization and G protein-independent ERK signaling were also abolished by point mutation at serine 348. In contrast, this mutant at serine residues had no demonstrable impact on apelin-13-induced G protein activation and its intracellular signaling. These findings suggest that mutation of serine 348 resulted in inactive GRK/β-arrestin. However, there was no change in the active G protein thus, APJ conformation was biased. These results provide important information on the molecular interplay and impact of the APJ function, which may be extrapolated to design novel drugs for cardiac hypertrophy based on this biased signal pathway. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Stress induces pain transition by potentiation of AMPA receptor phosphorylation.

Science.gov (United States)

Li, Changsheng; Yang, Ya; Liu, Sufang; Fang, Huaqiang; Zhang, Yong; Furmanski, Orion; Skinner, John; Xing, Ying; Johns, Roger A; Huganir, Richard L; Tao, Feng

2014-10-08

Chronic postsurgical pain is a serious issue in clinical practice. After surgery, patients experience ongoing pain or become sensitive to incident, normally nonpainful stimulation. The intensity and duration of postsurgical pain vary. However, it is unclear how the transition from acute to chronic pain occurs. Here we showed that social defeat stress enhanced plantar incision-induced AMPA receptor GluA1 phosphorylation at the Ser831 site in the spinal cord and greatly prolonged plantar incision-induced pain. Interestingly, targeted mutation of the GluA1 phosphorylation site Ser831 significantly inhibited stress-induced prolongation of incisional pain. In addition, stress hormones enhanced GluA1 phosphorylation and AMPA receptor-mediated electrical activity in the spinal cord. Subthreshold stimulation induced spinal long-term potentiation in GluA1 phosphomimetic mutant mice, but not in wild-type mice. Therefore, spinal AMPA receptor phosphorylation contributes to the mechanisms underlying stress-induced pain transition. Copyright © 2014 the authors 0270-6474/14/3413737-10$15.00/0.

Radiation-induced base substitution mutagenesis in single-stranded DNA phage M13

International Nuclear Information System (INIS)

Brandenburger, A.; Godson, G.N.; Glickman, B.W.; Sluis, C.A. van

1981-01-01

To elucidate the relative contributions of targeted and untargeted mutations to γ and UV radiation mutagenesis, the DNA sequences of 174 M13 revertant phages isolated from stocks of irradiated or unirradiated amber mutants grown in irradiated (SOS-induced) or unirradiated (non-induced) host bacteria, have been determined. Differences in the spectra of base change mutations induced in the various conditions were apparent, but no obvious specificity of mutagenesis was detected. In particular, under the present conditions, pyrimidine dimers did not seem to be the principal sites of UV-induced base substitution mutagenesis, suggesting that such mutagenesis occurs at the sites of lesions other than pyrimidine dimers, or is untargeted. (U.K.)

Training increases the concentration of [3H]ouabain-binding sites in rat skeletal muscle

DEFF Research Database (Denmark)

Kjeldsen, K; Richter, Erik; Galbo, H

1986-01-01

]ouabain-binding-site concentration in the diaphragm, but in the heart ventricles, the K+-dependent 3-O-methylfluorescein phosphatase activity increased by 20% (P less than 0.001). Muscle inactivity induced by denervation, plaster immobilisation or tenotomy reduced the [3H]ouabain-binding-site concentration by 20-30% (P less than 0...

Inducible nucleosome depletion at OREBP-binding-sites by hypertonic stress.

Directory of Open Access Journals (Sweden)

Edith H Y Tong

Full Text Available BACKGROUND: Osmotic Response Element-Binding Protein (OREBP, also known as TonEBP or NFAT5, is a unique transcription factor. It is hitherto the only known mammalian transcription factor that regulates hypertonic stress-induced gene transcription. In addition, unlike other monomeric members of the NFAT family, OREBP exists as a homodimer and it is the only transcription factor known to bind naked DNA targets by complete encirclement in vitro. Nevertheless, how OREBP interacts with target DNA, also known as ORE/TonE, and how it elicits gene transcription in vivo, remains unknown. METHODOLOGY: Using hypertonic induction of the aldose reductase (AR gene activation as a model, we showed that OREs contained dynamic nucleosomes. Hypertonic stress induced a rapid and reversible loss of nucleosome(s around the OREs. The loss of nucleosome(s was found to be initiated by an OREBP-independent mechanism, but was significantly potentiated in the presence of OREBP. Furthermore, hypertonic induction of AR gene was associated with an OREBP-dependent hyperacetylation of histones that spanned the 5' upstream sequences and at least some exons of the gene. Nevertheless, nucleosome loss was not regulated by the acetylation status of histone. SIGNIFICANCE: Our findings offer novel insights into the mechanism of OREBP-dependent transcriptional regulation and provide a basis for understanding how histone eviction and transcription factor recruitment are coupled.

Measuring site occupancy: a new perspective on cysteine oxidation.

Science.gov (United States)

Rogowska-Wrzesinska, Adelina; Wojdyla, Katarzyna; Williamson, James; Roepstorff, Peter

2014-10-01

Site occupancy is an extremely important aspect of quantification of protein modifications. Knowing the degree of modification of each oxidised cysteine residue is critical to understanding the biological role of these modifications. Yet modification site occupancy is very often overlooked, in part because there are very few analytical tools that allow such measurements. Here we present a new strategy, which provides quantitative analysis of cysteine S-nitrosylation (SNO) and S-sulfenylation (SOH) simultaneously at the resolution of single cysteine and allows for determination of relative oxidation occupancy of the modification site. We show that, on one hand, heavily modified cysteines are not necessarily involved in the response to oxidative stress. On the other hand residues with low modification level can be dramatically affected by mild oxidative imbalance. We make use of high resolution mass spectrometry. The method relies on differential reduction of "total" cysteines, SNO cysteines and SOH cysteines with TCEP, sodium ascorbate and sodium arsenite respectively followed by iodoTMT(TM) alkylation. Enrichment of iodoTMT(TM)-containing peptides is performed using anti-TMT antibody. In vivo model of mild oxidative stress in Escherichia coli is used. To induce endogenous SNO bacteria were grown anaerobically in minimal media supplemented with fumarate or nitrate. Short-term treatment with submilimolar levels of hydrogen peroxide were used to induce SOH. We have quantified 114 SNO/SOH modified peptides corresponding to 90 proteins. Only 6 modified peptides changed significantly under mild oxidative stress. Quantitative information allowed us to determine relative modification site occupancy of each identified modified residue and pin point heavily modified ones. The method proved to be precise and sensitive enough to detect and quantify endogenous levels of oxidative stress on proteome-wide scale and brings a new perspective on the role of the modification site

Activation of the Arabidopsis membrane-bound transcription factor bZIP28 is mediated by site-2 protease, but not site-1 protease.

Science.gov (United States)

Iwata, Yuji; Ashida, Makoto; Hasegawa, Chisa; Tabara, Kazuki; Mishiba, Kei-Ichiro; Koizumi, Nozomu

2017-08-01

The unfolded protein response (UPR) is a homeostatic cellular response conserved in eukaryotic cells to alleviate the accumulation of unfolded proteins in the endoplasmic reticulum (ER). Arabidopsis bZIP28 is a membrane-bound transcription factor activated by proteolytic cleavage in response to ER stress, thereby releasing its cytosolic portion containing the bZIP domain from the membrane to translocate into the nucleus where it induces the transcription of genes encoding ER-resident molecular chaperones and folding enzymes. It has been widely recognized that the proteolytic activation of bZIP28 is mediated by the sequential cleavage of site-1 protease (S1P) and site-2 protease (S2P). In the present study we provide evidence that bZIP28 protein is cleaved by S2P, but not by S1P. We demonstrated that wild-type and s1p mutant plants produce the active, nuclear form of bZIP28 in response to the ER stress inducer tunicamycin. In contrast, tunicamycin-treated s2p mutants do not accumulate the active, nuclear form of bZIP28. Consistent with these observations, s2p mutants, but not s1p mutants, exhibited a defective transcriptional response of ER stress-responsive genes and significantly higher sensitivity to tunicamycin. Interestingly, s2p mutants accumulate two membrane-bound bZIP28 fragments with a shorter ER lumen-facing C-terminal domain. Importantly, the predicted cleavage sites are located far from the canonical S1P recognition motif previously described. We propose that ER stress-induced proteolytic activation of bZIP28 is mediated by the sequential actions of as-yet-unidentified protease(s) and S2P, and does not require S1P. © 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd.

Risk Factors of Suicidal Ideations and Attempts in Talented, At-Risk Girls

Science.gov (United States)

Hull-Blanks, Elva E.; Kerr, Barbara A.; Robinson Kurpius, Sharon E.

2004-01-01

The purpose of the present study was to investigate the relationships among suicidality, substance use, self-esteem, family structure, and eight personality characteristics (harm avoidance, impulsivity, aggression, social recognition, cognitive structure, succorance, abasement, and achievement) with 337 talented, at-risk, adolescent girls. Results…

Alpha 4 integrin directs virus-activated CD8+ T cells to sites of infection

DEFF Research Database (Denmark)

Christensen, Jan Pravsgaard; Andersson, E C; Scheynius, A

1995-01-01

This article examines the role of VLA-4 in directing lymphocytes to sites of viral infection using the murine lymphocytic choriomeningitis virus infection (LCMV) as the model system. This virus by itself induces little or no inflammation, but in most mouse/virus strain combinations a potent T cell...... response is induced, which is associated with marked CD8+ cell-mediated inflammation. Two expressions of LCMV-induced inflammation were studied: meningitis induced by intracerebral infection and adoptive transfer of virus-specific delayed-type hypersensitivity. Our previous studies have shown that LCMV...

Endogenously generated plasmin at the vascular wall injury site amplifies lysine binding site-dependent plasminogen accumulation in microthrombi.

Directory of Open Access Journals (Sweden)

Tomasz Brzoska

Full Text Available The fibrinolytic system plays a pivotal role in the regulation of hemostasis; however, it remains unclear how and when the system is triggered to induce thrombolysis. Using intra-vital confocal fluorescence microscopy, we investigated the process of plasminogen binding to laser-induced platelet-rich microthrombi generated in the mesenteric vein of transgenic mice expressing green fluorescent protein (GFP. The accumulation of GFP-expressing platelets as well as exogenously infused Alexa Fluor 568-labeled Glu-plasminogen (Glu-plg on the injured vessel wall was assessed by measuring the increase in the corresponding fluorescence intensities. Glu-plg accumulated in a time-dependent manner in the center of the microthrombus, where phosphatidylserine is exposed on platelet surfaces and fibrin formation takes place. The rates of binding of Glu-plg in the presence of ε-aminocaproic acid and carboxypeptidase B, as well as the rates of binding of mini-plasminogen lacking kringle domains 1-4 and lysine binding sites, were significantly lower than that of Glu-plg alone, suggesting that the binding was dependent on lysine binding sites. Furthermore, aprotinin significantly suppressed the accumulation of Glu-plg, suggesting that endogenously generated plasmin activity is a prerequisite for the accumulation. In spite of the endogenous generation of plasmin and accumulation of Glu-plg in the center of microthrombi, the microthrombi did not change in size during the 2-hour observation period. When human tissue plasminogen activator was administered intravenously, Glu-plg further accumulated and the microthrombi were lysed. Glu-plg appeared to accumulate in the center of microthrombi in the early phase of microthrombus formation, and plasmin activity and lysine binding sites were required for this accumulation.

Effects of Edge on-Site Potential in a Honeycomb Topological Magnon Insulator

Science.gov (United States)

Pantaleón, Pierre A.; Xian, Yang

2018-06-01

While the deviation of the edge on-site potential from the bulk values in a magnonic topological honeycomb lattice leads to the formation of edge states in a bearded boundary, this is not the case for a zigzag termination, where no edge state is found. In a semi-infinite lattice, the intrinsic on-site interactions along the boundary sites generate an effective defect and this gives rise to Tamm-like edge states. If a nontrivial gap is induced, both Tamm-like and topologically protected edge states appear in the band structure. The effective defect can be strengthened by an external on-site potential, and the dispersion relation, velocity and magnon density of the edge states all become tunable.

Cryotherapy-induced milia en plaque: case report and literature review.

Science.gov (United States)

Beutler, Bryce David; Cohen, Philip R

2014-12-12

Cryotherapy-induced milia is a rarely described cutaneous reaction that may occur in patients who have received cryotherapy with liquid nitrogen. Cryotherapy-induced milia is characterized by 1-2 millimeter white dermal cysts that develop at the healed cryotherapy site. Milia en plaque, an erythematous plaque containing numerous milia, has not previously been described following treatment of a skin lesion with liquid nitrogen cryotherapy. We describe a man who developed cryotherapy-induced milia en plaque after receiving cryotherapy to his dorsal hand for the treatment of an actinic keratosis. We also summarize the potential complications of cryotherapy, the differential diagnosis of milia en plaque, and therapeutic interventions for this lesion. The features of a man with cryotherapy-induced milia en plaque are presented. Using PubMed, the following terms were searched and relevant citations assessed: cryosurgery, cryotherapy, hypothermia, milia, milia en plaque, and Wolf's isotopic response. In addition, the literature on cryotherapy-induced milia and cryotherapy-induced milia en plaque is reviewed. Our patient developed cryotherapy-induced milia en plaque shortly after his cryotherapy site had healed. Some of the asymptomatic cystic dermal lesions had spontaneously resolved when a lesional biopsy was performed to confirm the diagnosis. The diagnosis, natural course, and potential treatments were discussed with the patient. Subsequent management was to observe the area; at follow-up examination, the remainder of the milia had also spontaneously resolved. Cryotherapy-induced milia is a benign condition characterized by the development of small white dermal cystic lesions that develop at a healed liquid nitrogen cryotherapy site. The lesions may appear individually or as milia en plaque. While the mechanism of pathogenesis is unknown, we postulate that the condition is an example of Wolf's isotopic response, in which a new, unrelated skin disease develops at the

Spinal cord stimulation suppresses bradycardias and atrial tachyarrhythmias induced by mediastinal nerve stimulation in dogs.

Science.gov (United States)

Cardinal, René; Pagé, Pierre; Vermeulen, Michel; Bouchard, Caroline; Ardell, Jeffrey L; Foreman, Robert D; Armour, J Andrew

2006-11-01

Spinal cord stimulation (SCS) applied to the dorsal aspect of the cranial thoracic cord imparts cardioprotection under conditions of neuronally dependent cardiac stress. This study investigated whether neuronally induced atrial arrhythmias can be modulated by SCS. In 16 anesthetized dogs with intact stellate ganglia and in five with bilateral stellectomy, trains of five electrical stimuli were delivered during the atrial refractory period to right- or left-sided mediastinal nerves for up to 20 s before and after SCS (20 min). Recordings were obtained from 191 biatrial epicardial sites. Before SCS (11 animals), mediastinal nerve stimulation initiated bradycardia alone (12 nerve sites), bradycardia followed by tachyarrhythmia/fibrillation (50 sites), as well as tachyarrhythmia/fibrillation without a preceding bradycardia (21 sites). After SCS, the number of responsive sites inducing bradycardia was reduced by 25% (62 to 47 sites), and the cycle length prolongation in residual bradycardias was reduced. The number of responsive sites inducing tachyarrhythmia was reduced by 60% (71 to 29 sites). Once elicited, residual tachyarrhythmias arose from similar epicardial foci, displaying similar dynamics (cycle length) as in control states. In the absence of SCS, bradycardias and tachyarrhythmias induced by repeat nerve stimulation were reproducible (five additional animals). After bilateral stellectomy, SCS no longer influenced neuronal induction of bradycardia and atrial tachyarrhythmias. These data indicate that SCS obtunds the induction of atrial arrhythmias resulting from excessive activation of intrinsic cardiac neurons and that such protective effects depend on the integrity of nerves coursing via the subclavian ansae and stellate ganglia.

Crystal structure of the apurinic/apyrimidinic endonuclease IV from Mycobacterium tuberculosis.

Science.gov (United States)

Zhang, Wei; Xu, Yueyang; Yan, Mengrong; Li, Shanshan; Wang, Huiying; Yang, Haitao; Zhou, Weihong; Rao, Zihe

2018-03-25

Endonuclease IV is a typical endonuclease of the apurinic-apyrimidinic (AP) or abasic endonuclease superfamily. It repairs damaged DNA through base excision repair by cleaving the DNA backbone immediately 5' of an AP site. In Mycobacterium tuberculosis, endonuclease IV is the major AP endonuclease. This enzyme is absent from mammalian cells, making it an attractive target for anti-tuberculosis drug development. In this study, the structure of the recombinant endonuclease IV from M. tuberculosis (MtbEndo IV) was determined at a high resolution of 1.18 Å. MtbEndo IV was found to have a classical α8β8-fold TIM barrel with loops on its surface connecting the α-helices and β-strands that constitute a groove for DNA binding. Three zinc ions were identified at the active site. A comparison between the structures of MtbEndo IV and Escherichia coli End IV suggested that Gln32 of MtbEndo IV may plays a role in regulating substrate binding. Copyright © 2018. Published by Elsevier Inc.

Modification of DNA radiolysis by DNA-binding proteins: Structural aspects

International Nuclear Information System (INIS)

Davidkova, M.; Stisova, V.; Goffinont, S.; Gillard, N.; Castaing, B.; Spotheim-Maurizot, M.

2006-01-01

Formation of specific complexes between proteins and their cognate DNA modulates the yields and the location of radiation damage on both partners of the complex. The radiolysis of DNA-protein complexes is studied for: (1) the Escherichia coli lactose operator-repressor complex, (2) the complex between DNA bearing an analogue of an abasic site and the repair protein Fpg of Lactococcus lactis. Experimental patterns of DNA damages are presented and compared to predicted damage distribution obtained using an improved version of the stochastic model RADACK. The same method is used for predicting the location of damages on the proteins. At doses lower than a threshold that depends on the system, proteins protect their specific binding site on DNA while at high doses, the studied complexes are disrupted mainly through protein damage. The loss of binding ability is the functional consequence of the amino-acids modification by OH . radicals. Many of the most probably damaged amino acids are essential for the DNA-protein interaction and within a complex are protected by DNA. (authors)

Salivary glands act as mucosal inductive sites via the formation of ectopic germinal centers after site-restricted MCMV infection.

Science.gov (United States)

Grewal, Jasvir S; Pilgrim, Mark J; Grewal, Suman; Kasman, Laura; Werner, Phillip; Bruorton, Mary E; London, Steven D; London, Lucille

2011-05-01

We investigated the hypothesis that salivary gland inoculation stimulates formation of ectopic germinal centers (GCs), transforming the gland into a mucosal inductive site. Intraglandular infection of mice with murine cytomegalovirus (MCMV; control: UV-inactivated MCMV) induces salivary gland ectopic follicles comprising cognate interactions between CD4(+) and B220(+) lymphocytes, IgM(+) and isotype-switched IgG(+) and IgA(+) B cells, antigen presenting cells, and follicular dendritic cells. B cells coexpressed the GC markers GCT (57%) and GL7 (52%), and bound the lectin peanut agglutinin. Lymphoid follicles were characterized by a 2- to 3-fold increase in mRNA for CXCL13 (lymphoid neogenesis), syndecan-1 (plasma cells), Blimp-1 (plasma cell development/differentiation), and a 2- to 6-fold increase for activation-induced cytidine deaminase, PAX5, and the nonexcised rearranged DNA of an IgA class-switch event, supporting somatic hypermutation and class-switch recombination within the salivary follicles. Intraglandular inoculation also provided protection against a systemic MCMV challenge, as evidenced by decreased viral titers (10(5) plaque-forming units to undetectable), and restoration of normal salivary flow rates from a 6-fold decrease. Therefore, these features suggest that the salivary gland participates in oral mucosal immunity via generation of ectopic GCs, which function as ectopic mucosal inductive sites.

Mn fraction substitutional site and defects induced magnetism in Mn-implanted 6H-SiC

Energy Technology Data Exchange (ETDEWEB)

Bouziane, K., E-mail: Khalid.bouziane@uir.ac.ma [Pôle Energies Renouvelables et Etudes Pétrolières, Université Internationale de Rabat, 11000 – Salé el Jadida, Technopolis (Morocco); Al Azri, M.; Elzain, M. [Department of Physics, College of Science, Sultan Qaboos University, P.O. Box 36, Al-Khodh 123 (Oman); Chérif, S.M. [LSPM (CNRS-UPR 3407), Université Paris, 13-Nord, 99, Avenue Jean Baptiste Clément, 93430 Villetaneuse (France); Mamor, M. [Equipe MSISM, Faculté Poly-Disciplinaire, B.P. 4162 Safi, Université Cadi Ayyad, Marrakech (Morocco); Declémy, A. [Institut P’, CNRS – Université de Poitiers – ENSMA, UPR 3346, SP2MI – Téléport 2, 11 boulevard Marie et Pierre Curie, BP 30179, F-86962 Futuroscope Chasseneuil Cedex (France); Thomé, L. [CSNSM-Orsay, Bât. 108, Université d’Orsay, F-91405 Orsay (France)

2015-05-25

Highlights: • Shallow Mn-implanted 6H-SiC crystal. • Correlation between Mn-substitutional site concentration and magnetism. • Correlation between defects nature surrounding Mn site and magnetism. • Correlation of magnetism in Mn-doped SiC to Mn at Si sites and vacancy-related defect. - Abstract: n-type 6H-SiC (0 0 0 1) single crystal substrates were implanted with three fluences of manganese (Mn{sup +}) ions: 5 × 10{sup 15}, 1 × 10{sup 16} and 5 × 10{sup 16} cm{sup −2} with implantation energy of 80 keV at 365 °C to stimulate dynamic annealing. The samples were characterized using Rutherford backscattering channeling spectroscopy (RBS/C), high-resolution X-ray diffraction technique (HRXRD), and Superconducting Quantum Interference Device (SQUID) techniques. Two main defect regions have been identified using RBS/C spectra fitted with the McChasy code combined to SRIM simulations. Intermediate defects depth region is associated with vacancies (D{sub V}) and deeper defect (D{sub N}) essentially related to the Si and C interstitial defects. The defect concentration and the maximum perpendicular strain exhibit similar increasing trend with the Mn{sup +} fluence. Furthermore, the amount of Mn atoms at Si substitutional sites and the corresponding magnetic moment per Mn atom were found to increase with increasing Mn fluence from 0.7 μ{sub B} to 1.7 μ{sub B} and then collapsing to 0.2 μ{sub B}. Moreover, a strong correlation has been found between the magnetic moment and the combination of both large D{sub V}/D{sub N} ratio and high Mn at Si sites. These results are corroborated by our ab initio calculations considering the most stable configurations showing that besides the amount of Mn substituting Si sites, local vacancy-rich environment is playing a crucial role in enhancing the magnetism.

Local site effect of soil slope based on microtremor measurement in Samigaluh, Kulon Progo Yogyakarta

Science.gov (United States)

Prabowo, U. N.; Amalia, A. F.; Wiranata, F. E.

2018-03-01

This paper investigated soil slope-local site effect of earthquake inducing landslide by using microtremor Horizontal to Vertical Spectral Ratio (HVSR) method. Microtremor measurements of 15 sites which were recorded for 45 minutes at each site were carried out in Ngargosari village, Samigaluh, Kulon Progo-Indonesia. Microtremor analysis using HVSR method was performed using Geopsy software. HVSR method resulted in predominant frequency values that ranges between 2,77 to 13,82 Hz and amplification factors varied from 0,46 to 5,70. The predominant frequency is associated with the depth of bedrock and the amplification factor reflects the geological condition of soil (sedimentary layer). The soil vulnerability index (Kg) varied from 0,08 to 5,77 and the higher value (Kg>3,4) in the south of the research area was identified as the weak zone of earthquake inducing landslide.

An investigation of Laser Induced Breakdown Spectroscopy for use as a control in the laser removal of rock from fossils found at the Malapa hominin site, South Africa

Energy Technology Data Exchange (ETDEWEB)

Roberts, D.E., E-mail: troberts@csir.co.za [CSIR National Laser Centre, PO Box 395, Meiring Naude Road, Pretoria 0001 (South Africa); Plessis, A. du [CSIR National Laser Centre, PO Box 395, Meiring Naude Road, Pretoria 0001 (South Africa); University of Stellenbosch, Private Bag X1, Matieland, Stellenbosch (South Africa); Steyn, J.; Botha, L.R.; Pityana, S. [CSIR National Laser Centre, PO Box 395, Meiring Naude Road, Pretoria 0001 (South Africa); Berger, L.R. [Institute for Human Evolution, School of GeoSciences, University of Witwatersrand, Private Bag 3, Wits 2050 (South Africa)

2012-07-15

Laser Induced Breakdown Spectroscopy (LIBS) was used to study the spectra from fossils and surrounding rock recovered from the Cradle of Mankind site at Malapa, South Africa. The objective was to find a suitable spectral line(s), specific to fossils, which could be used as a control signal to limit damage to fossils during high speed laser removal of the encasing rock. The calcified clastic matrix (rock) encasing the fossils was found to emit a variety of complex LIBS spectra. Nevertheless, it was found possible to distinguish fossils in a single LIBS pulse, and without significant damage to the fossil, using spectral lines of neutral phosphorus. - Highlights: Black-Right-Pointing-Pointer LIBS used to discriminate fossils from rock as potential processing control mechanism. Black-Right-Pointing-Pointer 2 million year old fossils from Malapa hominin site found to be high in phosphorus. Black-Right-Pointing-Pointer Rock spectral lines from silicon, iron and manganese, but no phosphorus. Black-Right-Pointing-Pointer Holds great promise for process control in laser preparation of fossils. Black-Right-Pointing-Pointer Also promising for accurate identification of fossils at excavation sites.

[Effects of surface roughness of bone cements on histological characteristics of induced membranes].

Science.gov (United States)

Liu, Hai-Xiao; Xu, Hua-Zi; Zhang, Yu; Hu, Gang; Shen, Yue; Cheng, Xiao-Jie; Peng, Lei

2012-08-01

To explore surface roughness of bone cement and surround tissue on histological characteristic of induced membranes. Bone cements with smooth and rough surface were implanted in radius bone defect, intramuscular and subcutaneous sites of rabbits, and formed induced membranes. Membranes were obtained and stained (HE) 6 weeks later. Images of membrane tissue were obtained and analyzed with an automated image analysis system. Five histological parameters of membranes were measured with thickness,area,cell density,ECM density and microvessel density. Double factor variance analysis was used to evaluate the effect of the two factors on histological characteristics of induced membranes. Membranes can be induced by each kind of bone cement and at all the three tissue sites. In histological parameters of thickness,area and micro vessel,there were significant differences among the membranes induced at different tissue sites (P = 0.000, P = 0.000, P = 0.000); whereas, there were no significant differences in histological parameters of cell density and ECM density (P = 0.734, P = 0.638). In all five histological parameters of membranes, there were no significant differences between the membranes induced by bone cements with different surface roughness (P = 0.506, P = 0.185, P = 0.883, P = 0.093, P = 0.918). Surround tissue rather than surface roughness of bone cements can affect the histological characteristics of induced membranes. The fibrocystic number, vascularity, mechanical tension and micro motion of the surround tissue may be closely correlated with the histological characteristics of induced membranes.

Catalytic zinc site and mechanism of the metalloenzyme PR-AMP cyclohydrolase.

Science.gov (United States)

D'Ordine, Robert L; Linger, Rebecca S; Thai, Carolyn J; Davisson, V Jo

2012-07-24

The enzyme N(1)-(5'-phosphoribosyl) adenosine-5'-monophosphate cyclohydrolase (PR-AMP cyclohydrolase) is a Zn(2+) metalloprotein encoded by the hisI gene. It catalyzes the third step of histidine biosynthesis, an uncommon ring-opening of a purine heterocycle for use in primary metabolism. A three-dimensional structure of the enzyme from Methanobacterium thermoautotrophicum has revealed that three conserved cysteine residues occur at the dimer interface and likely form the catalytic site. To investigate the functions of these cysteines in the enzyme from Methanococcus vannielii, a series of biochemical studies were pursued to test the basic hypothesis regarding their roles in catalysis. Inactivation of the enzyme activity by methyl methane thiosulfonate (MMTS) or 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) also compromised the Zn(2+) binding properties of the protein inducing loss of up to 90% of the metal. Overall reaction stoichiometry and the potassium cyanide (KCN) induced cleavage of the protein suggested that all three cysteines were modified in the process. The enzyme was protected from DTNB-induced inactivation by inclusion of the substrate N(1)-(5'-phosphoribosyl)adenosine 5'-monophosphate; (PR-AMP), while Mg(2+), a metal required for catalytic activity, enhanced the rate of inactivation. Site-directed mutations of the conserved C93, C109, C116 and the double mutant C109/C116 were prepared and analyzed for catalytic activity, Zn(2+) content, and reactivity with DTNB. Substitution of alanine for each of the conserved cysteines showed no measurable catalytic activity, and only the C116A was still capable of binding Zn(2+). Reactions of DTNB with the C109A/C116A double mutant showed that C93 is completely modified within 0.5 s. A model consistent with these data involves a DTNB-induced mixed disulfide linkage between C93 and C109 or C116, followed by ejection of the active site Zn(2+) and provides further evidence that the Zn(2+) coordination site involves the

Alternative right ventricular pacing sites.

Science.gov (United States)

Łuciuk, Dariusz; Łuciuk, Marek; Gajek, Jacek

2015-01-01

The main adverse effect of chronic stimulation is stimulation-induced heart failure in case of ventricular contraction dyssynchrony. Because of this fact, new techniques of stimulation should be considered to optimize electrotherapy. One of these methods is pacing from alternative right ventricular sites. The purpose of this article is to review currently accumulated data about alternative sites of cardiac pacing. Medline and PubMed bases were used to search English and Polish reports published recently. Recent studies report a deleterious effect of long term apical pacing. It is suggested that permanent apical stimulation, by omitting physiological conduction pattern with His-Purkinie network, may lead to electrical and mechanical dyssynchrony of heart muscle contraction. In the long term this pathological situation can lead to severe heart failure and death. Because of this, scientists began to search for some alternative sites of cardiac pacing to reduce the deleterious effect of stimulation. Based on current accumulated data, it is suggested that the right ventricular outflow tract, right ventricular septum, direct His-bundle or biventricular pacing are better alternatives due to more physiological electrical impulse propagation within the heart and the reduction of the dyssynchrony effect. These methods should preserve a better left ventricular function and prevent the development of heart failure in permanent paced patients. As there is still not enough, long-term, randomized, prospective, cross-over and multicenter studies, further research is required to validate the benefits of using this kind of therapy. The article should pay attention to new sites of cardiac stimulation as a better and safer method of treatment.

The sequence specificity of UV-induced DNA damage in a systematically altered DNA sequence.

Science.gov (United States)

Khoe, Clairine V; Chung, Long H; Murray, Vincent

2018-06-01

The sequence specificity of UV-induced DNA damage was investigated in a specifically designed DNA plasmid using two procedures: end-labelling and linear amplification. Absorption of UV photons by DNA leads to dimerisation of pyrimidine bases and produces two major photoproducts, cyclobutane pyrimidine dimers (CPDs) and pyrimidine(6-4)pyrimidone photoproducts (6-4PPs). A previous study had determined that two hexanucleotide sequences, 5'-GCTC*AC and 5'-TATT*AA, were high intensity UV-induced DNA damage sites. The UV clone plasmid was constructed by systematically altering each nucleotide of these two hexanucleotide sequences. One of the main goals of this study was to determine the influence of single nucleotide alterations on the intensity of UV-induced DNA damage. The sequence 5'-GCTC*AC was designed to examine the sequence specificity of 6-4PPs and the highest intensity 6-4PP damage sites were found at 5'-GTTC*CC nucleotides. The sequence 5'-TATT*AA was devised to investigate the sequence specificity of CPDs and the highest intensity CPD damage sites were found at 5'-TTTT*CG nucleotides. It was proposed that the tetranucleotide DNA sequence, 5'-YTC*Y (where Y is T or C), was the consensus sequence for the highest intensity UV-induced 6-4PP adduct sites; while it was 5'-YTT*C for the highest intensity UV-induced CPD damage sites. These consensus tetranucleotides are composed entirely of consecutive pyrimidines and must have a DNA conformation that is highly productive for the absorption of UV photons. Crown Copyright © 2018. Published by Elsevier B.V. All rights reserved.

Site specific information in site selection

International Nuclear Information System (INIS)

Aeikaes, T.; Hautojaervi, A.

1998-01-01

The programme for the siting of a deep repository for final disposal of spent nuclear fuel was started already in 1983 and is carried out today by Posiva Oy which continues the work started by Teollisuuden Voima Oy. The programme aims at site selection by the end of the year 2000. The programme has progressed in successive interim stages with defined goals. After an early phase for site identification, five sites were selected in 1987 for preliminary site characterisation. Three of these were selected and judged to be best suited for the more detailed characterisation in 1992. An additional new site was included into the programme based on a separate feasibility study in the beginning of 1997. Since the year 1983 several safety assessments together with technical plans of the facility have been completed. When approaching the site selection the needs for more detailed consideration of the site specific properties in the safety assessment have been increased. The Finnish regulator STUK has published a proposal for general safety requirements for the final disposal of spent nuclear fuel in Finland. This set of requirements has been projected to be used in conjunction of the decision making by the end 2000. Based on the site evaluation all sites can provide a stable environment and there is evidence that the requirements for the longevity of the canister can be fulfilled at each site. In this manner the four candidate sites do not differ too much from each other. The main difference between the sites is in the salinity of the deep groundwater. The significance of differences in the salinity for the long-term safety cannot be defined yet. The differences may contribute to the discussion of the longevity of the bentonite buffer and also to the modelling of the groundwater flow and transport. The use of the geosphere as a transport barrier is basically culminated on the questions about sparse but fast flow routes and 'how bad channeling can be'. To answer these questions

Fluorescent-light-induced lethality and DNA repair in normal and xeroderma pigmentosum fibroblasts

International Nuclear Information System (INIS)

Ritter, M.A.; Williams, J.R.

1981-01-01

Cell survival and induction of endonuclease-sensitive sites in DNA were measured in human fibroblast cells exposed to fluorescent light or germicidal ultraviolet light. Cells from a xeroderma pigmentosum patient were hypersensitive to cell killing by fluorescent light, although less so than for germicidal ultraviolet light. Xeroderma pigmentosum cells were deficient in the removal of fluorescent light-induced endonuclease sites that are probably pyrimidine dimers, and both the xeroderma pigmentosum and normal cells removed these sites with kinetics indistinguishable from those for ultraviolet light-induced sites. A comparison of fluorescent with ultraviolet light data demonstrates that there are markedly fewer pyrimidine dimers per lethal event for fluorescent than for ultraviolet light, suggesting a major role for non-dimer damage in fluorescent lethality. (Auth.)

Induced-fit Mechanism for Prolyl Endopeptidase

Energy Technology Data Exchange (ETDEWEB)

Li, Min; Chen, Changqing; Davies, David R.; Chiu, Thang K. (NIH); (LSU); (Chinese Aca. Sci.)

2010-11-15

Prolyl peptidases cleave proteins at proline residues and are of importance for cancer, neurological function, and type II diabetes. Prolyl endopeptidase (PEP) cleaves neuropeptides and is a drug target for neuropsychiatric diseases such as post-traumatic stress disorder, depression, and schizophrenia. Previous structural analyses showing little differences between native and substrate-bound structures have suggested a lock-and-key catalytic mechanism. We now directly demonstrate from seven structures of Aeromonus punctata PEP that the mechanism is instead induced fit: the native enzyme exists in a conformationally flexible opened state with a large interdomain opening between the {beta}-propeller and {alpha}/{beta}-hydrolase domains; addition of substrate to preformed native crystals induces a large scale conformational change into a closed state with induced-fit adjustments of the active site, and inhibition of this conformational change prevents substrate binding. Absolute sequence conservation among 28 orthologs of residues at the active site and critical residues at the interdomain interface indicates that this mechanism is conserved in all PEPs. This finding has immediate implications for the use of conformationally targeted drug design to improve specificity of inhibition against this family of proline-specific serine proteases.

Spectroscopic link between adsorption site occupation and local surface chemical reactivity

DEFF Research Database (Denmark)

Baraldi, A.; Lizzit, S.; Comelli, G.

2004-01-01

rules, from which adsorption sites are directly determined. Theoretical calculations rationalize the results for transition metal surfaces in terms of the energy shift of the d-band center of mass and this proves that adsorbate-induced SCL shifts provide a spectroscopic measure of local surface...

Base-repair substitutions alter the site-specific mutagenicity of UV and MNNG in the SUP4-o gene of the yeast

International Nuclear Information System (INIS)

Kunz, B.A.; Ayre, B.G.; Downes, A.M.T.; Kohalmi, S.E.; McMaster, C.R.; Peters, M.G.

1989-01-01

Yeast strains carrying SUP4-ogenes that habe base-pair substitutions at hotspots for UV or MNNG mutagenesis were treated with these agents. In both cases, the induced mutation frequencies were substantially reduced. Furthermore, specific substitutions at positions in SUP4-o that had not been mutated by MNNG resulted in the recovery of MNNG-induced mutations at these sites. These results demonstrate that base-pair identity is an important factor determining the site-specific mutagenicity of UV and MNNG in yeast. For UV, our findings suggest that the type of lesion that is induced, but not flanking DNA sequences, plays a role in specifying mutability at the sites examined. In contrast, DNA sequence context seems to be an important factor for MNNG mutagenesis. (author). 19 refs.; 3 tabs

Subsurface imaging of water electrical conductivity, hydraulic permeability and lithology at contaminated sites by induced polarization

DEFF Research Database (Denmark)

Maurya, P. K.; Balbarini, Nicola; Møller, I.

2018-01-01

At contaminated sites, knowledge about geology and hydraulic properties of the subsurface and extent of the contamination is needed for assessing the risk and for designing potential site remediation. In this study, we have developed a new approach for characterizing contaminated sites through time...... geological logs. On average the IP-derived and measured permeability values agreed within one order of magnitude, except for those close to boundaries between lithological layers (e.g. between sand and clay), where mismatches occurred due to the lack of vertical resolution in the geophysical imaging...

Removal of N-linked glycosylations at acidic pH by PNGase a facilitates hydrogen/deuterium exchange mass spectrometry analysis of N-Linked glycoproteins

DEFF Research Database (Denmark)

Jensen, Pernille Foged; Comamala, Gerard; Trelle, Morten Beck

2016-01-01

for analysis of the conformational dynamics of N-linked glycoproteins that utilizes the enzyme PNGase A for deglycosylation of labeled peptic N-linked glycopeptides at HDX quench conditions, i.e., acidic pH and low temperature. PNGase A-based deglycosylation is thus performed after labeling (post...

Induced Seismicity at the UK "Hot Dry Rock" Test Site for Geothermal Energy Production

OpenAIRE

Li, Xun; Main, Ian; Jupe, Andrew

2018-01-01

In enhanced geothermal systems (EGS), fluid is injected at high pressure in order to stimulate fracturing and/or fluid flow through otherwise relatively impermeable underlying hot rocks to generate power and/or heat. The stimulation induces micro-earthquakes whose precise triggering mechanism and relationship to new and pre-existing fracture networks are still the subject of some debate. Here we analyse the dataset for induced micro-earthquakes at the UK “hot dry rock” experimental geothermal...

Occurrence and elimination of sites sensitive to UV-endonuclease in UV-irradiated E. coli cells

Energy Technology Data Exchange (ETDEWEB)

Kleibl, K; Sedliakova, M [Slovenska Akademia Vied, Bratislava (Czechoslovakia). Vyskumny Ustav Onkologicky

1979-01-01

The occurrence and elimination of sites sensitive to the endonucleolytic action of crude extract from M. luteus (Es sites) were studied in both the parental and daughter DNA of E. coli B/r Hcr/sup +/ irradiated either with lethal fluence only (LF) or with inducing and lethal fluence (IF+LF); after the lethal fluence protein synthesis could either take place or it was inhibited by chlorampehnicol (CAP). The data obtained showed that in the wild type UV-irradiated cells Es sites could be eliminated from their DNA molecules either through pyrimidine dimer excision or through the modification of dimers on replication. It appears that DNA repair takes place most efficiently in cells irradiated with IF+LF and postincubated with CAP; in these conditions cells are supplied with inducible proteins, and enough time for DNA repair is provided before the division of irradiated cells is resumed.

D-penicillamine induced degenerative dermopathy

Directory of Open Access Journals (Sweden)

Sujay Khandpur

2015-01-01

Full Text Available D-penicillamine interferes with elastin and collagen metabolism and produces several cutaneous and multi-systemic side-effects. We present two cases of Wilson′s disease who on long-term penicillamine therapy developed drug-induced degenerative dermopathy manifesting as skin fragility over pressure sites and cutis laxa-like changes.

Site organization and site arrangement

International Nuclear Information System (INIS)

Boissonnet, B.; Macqueron, J.F.

1976-01-01

The present paper deals with criteria for the choice of a production unit or power plant site, the organization and development of a site in terms of its particular characteristics and takes into account personnel considerations in site organizations as well as the problem of integrating the architecture into the environment. (RW) [de

UV light-induced cyclobutane pyrimidine dimers are mutagenic in mammalian cells

International Nuclear Information System (INIS)

Protic-Sabljic, M.; Tuteja, N.; Munson, P.J.; Hauser, J.; Kraemer, K.H.; Dixon, K.

1986-01-01

We used a simian virus 40-based shuttle vector plasmid, pZ189, to determine the role of pyrimidine cyclobutane dimers in UV light-induced mutagenesis in monkey cells. The vector DNA was UV irradiated and then introduced into monkey cells by transfection. After replication, vector DNA was recovered from the cells and tested for mutations in its supF suppressor tRNA marker gene by transformation of Escherichia coli carrying a nonsense mutation in the beta-galactosidase gene. When the irradiated vector was treated with E. coli photolyase prior to transfection, pyrimidine cyclobutane dimers were removed selectively. Removal of approximately 90% of the pyrimidine cyclobutane dimers increased the biological activity of the vector by 75% and reduced its mutation frequency by 80%. Sequence analysis of 72 mutants recovered indicated that there were significantly fewer tandem double-base changes and G X C----A X T transitions (particularly at CC sites) after photoreactivation of the DNA. UV-induced photoproducts remained (although at greatly reduced levels) at all pyr-pyr sites after photoreactivation, but there was a relative increase in photoproducts at CC and TC sites and a relative decrease at TT and CT sites, presumably due to a persistence of (6-4) photoproducts at some CC and TC sites. These observations are consistent with the fact that mutations were found after photoreactivation at many sites at which only cyclobutane dimers would be expected to occur. From these results we conclude that UV-induced pyrimidine cyclobutane dimers are mutagenic in DNA replicated in monkey cells

Bronchial or Laryngeal Obstruction Induced by Exercise?

Directory of Open Access Journals (Sweden)

Ayoub Bey

2017-06-01

Full Text Available A child suspected of exercise-induced laryngeal obstruction and asthma is examined by laryngoscopy and respiratory resistance (Rrs after exercise challenge. Immediately at exercise cessation, the visualized adduction of the larynx in inspiration is reflected in a paroxystic increase in Rrs. While normal breathing has apparently resumed later on during recovery from exercise, the pattern of Rrs in inspiration is observed to reoccur following a deep breath or swallowing. The procedure may thus help diagnosing the site of exercise-induced obstruction when laryngoscopy is not available and identify re-inducers of laryngeal dysfunction.

Energy transfer at the active sites of heme proteins

International Nuclear Information System (INIS)

Dlott, D.D.; Hill, J.R.

1995-01-01

Experiments using a picosecond pump-probe apparatus at the Picosecond Free-electron Laser Center at Stanford University, were performed to investigate the relaxation of carbon monoxide bound to the active sites of heme proteins. The significance of these experiments is two-fold: (1) they provide detailed information about molecular dynamics occurring at the active sites of proteins; and (2) they provide insight into the nature of vibrational relaxation processes in condensed matter. Molecular engineering is used to construct various molecular systems which are studied with the FEL. We have studied native proteins, mainly myoglobin obtained from different species, mutant proteins produced by genetic engineering using recombinant DNA techniques, and a variety of model systems which mimic the structures of the active sites of native proteins, which are produced using molecular synthesis. Use of these different systems permits us to investigate how specific molecular structural changes affect dynamical processes occurring at the active sites. This research provides insight into the problems of how different species needs are fulfilled by heme proteins which have greatly different functionality, which is induced by rather small structural changes

Investigation of anatomical anomalies in Hanford Site mule deer

Energy Technology Data Exchange (ETDEWEB)

Tiller, B.L.; Cadwell, L.L.; Poston, T.M. [and others

1997-03-01

Rocky Mountain mule deer (Odocoileus hemionus hemionus), common residents of the Hanford Site, are an important part of the shrub-steppe ecosystem as well as being valued for aesthetics and hunting. Because mule deer have been protected from hunting on the Site for 50 years, the herd has developed unique population characteristics, including a large number of old animals and males with either large or atypically developed antlers, in contrast to other herds in the semi-arid regions of the Northwest. Hanford Site mule deer have been studied since 1991 because of the herd`s unique nature and high degree of public interest. A special study of the mule deer herd was initiated in 1993 after observations were made of a relatively large number of male deer with atypical, velvet-covered antlers. This report specifically describes our analyses of adult male deer found on the Site with atypical antlers. The report includes estimates of population densities and composition; home ranges, habitat uses, and dietary habits; natural and human-induced causes of mortality; and the herd`s overall health and reproductive status.

Investigation of anatomical anomalies in Hanford Site mule deer

International Nuclear Information System (INIS)

Tiller, B.L.; Cadwell, L.L.; Poston, T.M.

1997-03-01

Rocky Mountain mule deer (Odocoileus hemionus hemionus), common residents of the Hanford Site, are an important part of the shrub-steppe ecosystem as well as being valued for aesthetics and hunting. Because mule deer have been protected from hunting on the Site for 50 years, the herd has developed unique population characteristics, including a large number of old animals and males with either large or atypically developed antlers, in contrast to other herds in the semi-arid regions of the Northwest. Hanford Site mule deer have been studied since 1991 because of the herd's unique nature and high degree of public interest. A special study of the mule deer herd was initiated in 1993 after observations were made of a relatively large number of male deer with atypical, velvet-covered antlers. This report specifically describes our analyses of adult male deer found on the Site with atypical antlers. The report includes estimates of population densities and composition; home ranges, habitat uses, and dietary habits; natural and human-induced causes of mortality; and the herd's overall health and reproductive status

The Earliest Chinese Proto-Porcelain Excavated from Kiln Sites: An Elemental Analysis

OpenAIRE

Li, Yu; Zhang, Bin; Cheng, Huansheng; Zheng, Jianming

2015-01-01

In June 2012, the Piaoshan kiln site was excavated in Huzhou, Zhejiang Province, which hitherto proved to be the earliest known Chinese proto-porcelain kiln. Judging from the decorative patterns of unearthed impressed stoneware and proto-porcelain sherds, the site was determined to date to the late Xia (c. 2070-c. 1600 BC), the first dynasty of China. Here, we report on proton-induced X-ray emission analyses of 118 proto-porcelain and 35 impressed stoneware sherds from Piaoshan and five subse...

Interpreting sperm DNA damage in a diverse range of mammalian sperm by means of the two-tailed comet assay

Science.gov (United States)

Cortés-Gutiérrez, Elva I.; López-Fernández, Carmen; Fernández, José Luis; Dávila-Rodríguez, Martha I.; Johnston, Stephen D.; Gosálvez, Jaime

2014-01-01

Key Concepts The two-dimensional Two-Tailed Comet assay (TT-comet) protocol is a valuable technique to differentiate between single-stranded (SSBs) and double-stranded DNA breaks (DSBs) on the same sperm cell.Protein lysis inherent with the TT-comet protocol accounts for differences in sperm protamine composition at a species-specific level to produce reliable visualization of sperm DNA damage.Alkaline treatment may break the sugar–phosphate backbone in abasic sites or at sites with deoxyribose damage, transforming these lesions into DNA breaks that are also converted into ssDNA. These lesions are known as Alkali Labile Sites “ALSs.”DBD–FISH permits the in situ visualization of DNA breaks, abasic sites or alkaline-sensitive DNA regions.The alkaline comet single assay reveals that all mammalian species display constitutive ALS related with the requirement of the sperm to undergo transient changes in DNA structure linked with chromatin packing.Sperm DNA damage is associated with fertilization failure, impaired pre-and post- embryo implantation and poor pregnancy outcome.The TT is a valuable tool for identifying SSBs or DSBs in sperm cells with DNA fragmentation and can be therefore used for the purposes of fertility assessment. Sperm DNA damage is associated with fertilization failure, impaired pre-and post- embryo implantation and poor pregnancy outcome. A series of methodologies to assess DNA damage in spermatozoa have been developed but most are unable to differentiate between single-stranded DNA breaks (SSBs) and double-stranded DNA breaks (DSBs) on the same sperm cell. The two-dimensional Two-Tailed Comet assay (TT-comet) protocol highlighted in this review overcomes this limitation and emphasizes the importance in accounting for the difference in sperm protamine composition at a species-specific level for the appropriate preparation of the assay. The TT-comet is a modification of the original comet assay that uses a two dimensional electrophoresis to

The role of nicotinic acetylcholine and opioid systems of the ventral orbital cortex in modulation of formalin-induced orofacial pain in rats.

Science.gov (United States)

Yousofizadeh, Shahnaz; Tamaddonfard, Esmaeal; Farshid, Amir Abbas

2015-07-05

Nicotinic acetylcholine and opioid receptors are involved in modulation of pain. In the present study, we investigated the effects of microinjection of nicotinic acetylcholine and opioid compounds into the ventral orbital cortex (VOC) on the formalin-induced orofacial pain in rats. For this purpose, two guide cannulas were placed into the left and right sides of the VOC of the brain. Orofacial pain was induced by subcutaneous injection of a diluted formalin solution (50μl, 1.5%) into the right vibrissa pad and face rubbing durations were recorded at 3-min blocks for 45min. Formalin produced a marked biphasic pain response (first phase: 0-3min and second phase: 15-33min). Epibatidine (a nicotinic receptor agonist) at doses of 0.05, 0.1 and 0.2μg/site, morphine (an opioid receptor agonist) at doses of 0.5, 1 and 2μg/site and their sub-analgesic doses (0.025μg/site epibatidine with 0.25μg/site morphine) combination treatment suppressed the second phase of pain. The antinociceptive effect induced by 0.2μg/site of epibatidine, but not morphine (2μg/site), was prevented by 2μg/site of mecamylamine (a nicotinic receptor antagonist). Naloxone (an opioid receptor antagonist) at a dose of 2μg/site prevented the antinociceptive effects induced by 2μg/site of morphine and 0.2μg/site of epibatidine. No above-mentioned chemical compounds affected locomotor activity. These results showed that at the VOC level, epibatidine and morphine produced antinociception. In addition, opioid receptor might be involved in epibatidine-induced antinociception, but the antinociception induced by morphine was not mediated through nicotinic acetylcholine receptor. Copyright © 2015 Elsevier B.V. All rights reserved.

PHWR safety: design, siting and construction

International Nuclear Information System (INIS)

Sharma, V.K.

2002-01-01

In all activities associated with NPPs viz. siting, design, construction, commissioning and operation, safety is given overriding importance. The safety design principles of PHWRs are based on defence-in-depth approach, physical and functional separation between process and safety systems and also among various safety systems, redundancy to meet single failure criteria and postulation of a number of design basis events for which the plant must be designed. Apart from engineered safety systems, PHWRs have inherent characteristics which contribute to safety. In siting of a NPP, it is required to ensure that the given site does not pose undue radiological hazard to public and the environment both during normal operation as well as during and following an accident condition. For this purpose, all site related external events, both natural and man induced, are assessed for their effect on the plant and are considered as part of the design basis. Possible radiological impact of the NPP on environment and surrounding population is assessed and ensured to be within acceptable limits. During construction phase, it is essential that the NPP be built in accordance with design intent and with required quality of workmanship to ensure that the NPP will remain safe during all states of operation. This is achieved through careful execution and QA activities encompassing all aspects of component fabrication at manufacturer works, civil construction, site erection, assembly, and commissioning. Future trends in nuclear safety will continue to be based on existing principles which have proved to be sound. These will be further strengthened by features such as increasing use of passive means of performing safety functions and a more explicit treatment of severe accidents. (author)

Effects of Low-Level Autonomic Stimulation on Prevention of Atrial Fibrillation Induced by Acute Electrical Remodeling

Directory of Open Access Journals (Sweden)

Yubi Lin

2013-01-01

Full Text Available Background. Rapid atrial pacing (RAP can induce electrical and autonomic remodeling and facilitate atrial fibrillation (AF. Recent reports showed that low-level vagosympathetic nerve stimulation (LLVNS can suppress AF, as an antiarrhythmic effect. We hypothesized that LLVNS can reverse substrate heterogeneity induced by RAP. Methods and Results. Mongrel dogs were divided into (LLVNS+RAP and RAP groups. Electrode catheters were sutured to multiple atrial sites, and LLVNS was applied to cervical vagosympathetic trunks with voltage 50% below the threshold slowing sinus rate by ⩽30 msec. RAP induced a significant decrease in effective refractory period (ERP and increase in the window of vulnerability at all sites, characterized by descending and elevated gradient differences towards the ganglionic plexi (GP sites, respectively. The ERP dispersion was obviously enlarged by RAP and more significant when the ERP of GP-related sites was considered. Recovery time from AF was also prolonged significantly as a result of RAP. LLVNS could reverse all these changes induced by RAP and recover the heterogeneous substrate to baseline. Conclusions. LLVNS can reverse the electrical and autonomic remodeling and abolish the GP-central gradient differences induced by RAP, and thus it can recover the homogeneous substrate, which may be the underlying mechanism of its antiarrhythmic effect.

Geological-Hydrological Site Evaluation for NPP Planning

Energy Technology Data Exchange (ETDEWEB)

Faust, Brigitte; Mini, Paolo [Nordostsschweizerische Kraftwerke AG NOK, Parkstrasse 23, 5401 Baden (Switzerland)

2008-07-01

NOK is investigating the potential replacement of the current NPP in Beznau. In order to meet the requirements with respect to a general licence application, geological, seismological, and geotechnical engineering, but also hydrological boundary conditions have been defined. These conditions define the nature of necessary investigations to obtain the geological, seismic, geotechnical and hydrological data themselves forming the basis to determine the site suitability. Viability has to be provided that a NPP can be built and operated at the proposed site without compromising public health, safety and environment. The collected data are also the basis for the design of all safety relevant structures, systems, and components. For example, the latter have to withstand the effects of natural phenomena such as earthquakes and human induced impact such as airplane crash without loosing their capability to perform the assigned safety functions. (authors)

Geological-Hydrological Site Evaluation for NPP Planning

International Nuclear Information System (INIS)

Faust, Brigitte; Mini, Paolo

2008-01-01

NOK is investigating the potential replacement of the current NPP in Beznau. In order to meet the requirements with respect to a general licence application, geological, seismological, and geotechnical engineering, but also hydrological boundary conditions have been defined. These conditions define the nature of necessary investigations to obtain the geological, seismic, geotechnical and hydrological data themselves forming the basis to determine the site suitability. Viability has to be provided that a NPP can be built and operated at the proposed site without compromising public health, safety and environment. The collected data are also the basis for the design of all safety relevant structures, systems, and components. For example, the latter have to withstand the effects of natural phenomena such as earthquakes and human induced impact such as airplane crash without loosing their capability to perform the assigned safety functions. (authors)

First-principles study on ferromagnetism in double perovskite Sr2AlTaO6 doped with Cu or Zn at B sites

Science.gov (United States)

Li, Y. D.; Wang, C. C.; Guo, Y. M.; Yu, Y.; Lu, Q. L.; Huang, S. G.; Li, Q. J.; Wang, H.; Cheng, R. L.; Liu, C. S.

2018-05-01

The possibilities of ferromagnetism induced by nonmagnetic dopants (Cu, Zn) in double perovskite Sr2AlTaO6 at B sites are investigated by density functional theory. Calculations reveal that substitutions at Ta-site tend to form high spin electronic configurations and could induce ferromagnetism which can be attributed to the hole-mediated p- d hybridization between Cu (or Zn) eg states and the neighboring O 2p states. The dopants preferably substitute at Al-site and adopt low spin electronic structures. Due to the smaller hole concentration and weaker covalent intensity, Sr2AlTaO6 with dopants at Al-site exhibits p-type metallic semiconductors without spin polarization.

GLYX-13, a NMDA receptor glycine-site functional partial agonist, induces antidepressant-like effects without ketamine-like side effects.

Science.gov (United States)

Burgdorf, Jeffrey; Zhang, Xiao-lei; Nicholson, Katherine L; Balster, Robert L; Leander, J David; Stanton, Patric K; Gross, Amanda L; Kroes, Roger A; Moskal, Joseph R

2013-04-01

Recent human clinical studies with the NMDA receptor (NMDAR) antagonist ketamine have revealed profound and long-lasting antidepressant effects with rapid onset in several clinical trials, but antidepressant effects were preceded by dissociative side effects. Here we show that GLYX-13, a novel NMDAR glycine-site functional partial agonist, produces an antidepressant-like effect in the Porsolt, novelty induced hypophagia, and learned helplessness tests in rats without exhibiting substance abuse-related, gating, and sedative side effects of ketamine in the drug discrimination, conditioned place preference, pre-pulse inhibition and open-field tests. Like ketamine, the GLYX-13-induced antidepressant-like effects required AMPA/kainate receptor activation, as evidenced by the ability of NBQX to abolish the antidepressant-like effect. Both GLYX-13 and ketamine persistently (24 h) enhanced the induction of long-term potentiation of synaptic transmission and the magnitude of NMDAR-NR2B conductance at rat Schaffer collateral-CA1 synapses in vitro. Cell surface biotinylation studies showed that both GLYX-13 and ketamine led to increases in both NR2B and GluR1 protein levels, as measured by Western analysis, whereas no changes were seen in mRNA expression (microarray and qRT-PCR). GLYX-13, unlike ketamine, produced its antidepressant-like effect when injected directly into the medial prefrontal cortex (MPFC). These results suggest that GLYX-13 produces an antidepressant-like effect without the side effects seen with ketamine at least in part by directly modulating NR2B-containing NMDARs in the MPFC. Furthermore, the enhancement of 'metaplasticity' by both GLYX-13 and ketamine may help explain the long-lasting antidepressant effects of these NMDAR modulators. GLYX-13 is currently in a Phase II clinical development program for treatment-resistant depression.

Ab initio prediction of mutation-induced cryptic splice-site activation and exon skipping

Czech Academy of Sciences Publication Activity Database

Divina, Petr; Kvitkovicova, Andrea; Buratti, E.; Vorechovsky, I.

2009-01-01

Roč. 17, č. 6 (2009), s. 759-765 ISSN 1018-4813 Institutional research plan: CEZ:AV0Z50520514 Keywords : mutation * cryptic splice site * exon skipping Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.564, year: 2009

Imatinib mesylate is effective in children with chronic myelogenous leukemia in late chronic and advanced phase and in relapse after stem cell transplantation

NARCIS (Netherlands)

Millot, F; Guilhot, J; Nelken, B; Leblanc, T; De Bont, ES; Bekassy, AN; Gadner, H; Sufliarska, S; Stary, J; Gschaidmeier, H; Guilhot, F; Suttorp, M

A multicentric phase 2 study was conducted to determine the efficiency and the tolerance of imatinib mesylate in children with chronic myelogenous leukemia (CML) in advanced phase of the disease, in relapse after stem cell transplantation, or in case of failure to an interferon a-based regimen. In

Hydrazine and hydroxylamine as probes for O2-reduction site of mitochondrial cytochrome c oxidase.

Science.gov (United States)

Kubota, T; Yoshikawa, S

1993-01-01

Reactions of hydrazine and hydroxylamine with bovine heart cytochrome c oxidase in the fully reduced state were investigated under anaerobic conditions following the visible-Soret spectral change. Hydrazine gave a sharp band at 575 nm with 20% decrease in the alpha band at 603 nm, and hydroxylamine induced a 2 nm blue-shift for the alpha band without any clear splitting. The Soret band at 443 nm was decreased significantly in intensity, with the concomitant appearance of a shoulder with hydrazine or a peak with hydroxylamine, both near 430 nm. The dependence on pH of the affinity of these reagents for the enzyme indicates that only the deprotonated forms of these reagents bind to the enzyme, suggesting a highly hydrophobic environment of the haem ligand-biding site. These spectral changes were largely removed by addition of cyanide or CO. However, detailed analysis of these spectral changes indicates that hydrazine perturbs the shape of the spectral change induced by cyanide and hydroxylamine perturbs that induced by CO. These results suggest that these aldehyde reagents bind to haem a3 iron as well as to a second site which is most likely to be the formyl group on the haem periphery, and that these two sites bind these reagents anti-cooperatively with each other. PMID:8389138

Provocative Endoscopy to Identify Bleeding Site in Upper Gastrointestinal Bleeding: A Novel Approach in Transarterial Embolization.

Science.gov (United States)

Kamo, Minobu; Fuwa, Sokun; Fukuda, Katsuyuki; Fujita, Yoshiyuki; Kurihara, Yasuyuki

2016-07-01

This report describes a novel approach to endoscopically induce bleeding by removing a clot from the bleeding site during angiography for upper gastrointestinal (UGI) hemorrhage. This procedure enabled accurate identification of the bleeding site, allowing for successful targeted embolization despite a negative initial angiogram. Provocative endoscopy may be a feasible and useful option for angiography of obscure bleeding sites in patients with UGI arterial hemorrhage. Copyright © 2016 SIR. Published by Elsevier Inc. All rights reserved.

Longterm effects of cardiac mediastinal nerve cryoablation on neural inducibility of atrial fibrillation in canines.

Science.gov (United States)

Leiria, Tiago Luiz Luz; Glavinovic, Tamara; Armour, J Andrew; Cardinal, René; de Lima, Gustavo Glotz; Kus, Teresa

2011-04-26

In canines, excessive activation of select mediastinal nerve inputs to the intrinsic cardiac nervous system induces atrial fibrillation (AF). Since ablation of neural elements is proposed as an adjunct to circumferential pulmonary vein ablation for AF, we investigated the short and long-term effects of mediastinal nerve ablation on AF inducibility. Under general anesthesia, in 11 dogs several mediastinal nerve sites were identified on the superior vena cava that, when stimulated electrically during the atrial refractory period, reproducibly initiated AF. Cryoablation of one nerve site was then performed and inducibility retested early (1-2 months post Cryo; n=7) or late (4 months post Cryo; n=4). Four additional dogs that underwent a sham procedure were retested 1 to 2 months post-surgery. Stimulation induced AF at 91% of nerve sites tested in control versus 21% nerve sites early and 54% late post-ablation (both P<0.05). Fewer stimuli were required to induce AF in controls versus the Early Cryo group; this capacity returned to normal values in the Late Cryo group. AF episodes were longer in control versus the Early or Late Cryo groups. Heart rate responses to vagal or stellate ganglion stimulation, as well as to local nicotine infusion into the right coronary artery, were similar in all groups. In conclusion, focal damage to intrinsic cardiac neuronal inputs causes short-term stunning of neuronal inducibility of AF without major loss of overall adrenergic or cholinergic efferent neuronal control. That recovery of AF inducibility occurs rapidly post-surgery indicates the plasticity of intrathoracic neuronal elements to focal injury. Copyright © 2011 Elsevier B.V. All rights reserved.

Investigation of Trap Sites and Their Roles in Organic Triphenylamine-Based Photorefractive Materials

OpenAIRE

Tsujimura, Sho

2016-01-01

Organic photorefractive (PR) materials have been studied during the last quarter-century, and they have recently received much attention due to their updatable features that allow them to be used in dynamic holographic devices. However, understanding bulk trap sites that drive the PR effect (by inducing a space-charge field) remains a critical issue. In general, the trap site behavior can be controlled from the energetic point of view; however, bulk devices contain not only the energy trap si...

Impaired recovery of brain muscarinic receptor sites following an adaptive down-regulation induced by repeated administration of diisopropyl fluorophosphate in aged rats

International Nuclear Information System (INIS)

Pintor, A.; Fortuna, S.; De Angelis, S.; Michalek, H.

1990-01-01

Potential age-related differences in the recovery rate of brain cholinesterase activity (ChE) and muscarinic acetylcholine receptor binding sites (mAChRs) following reduction induced by repeated treatment with diisopropyl fluorophosphate (DFP) were evaluated in Sprague-Dawley rats. Male 3- and 24-month old rats were s.c. injected with DFP on alternate days for 2 weeks and killed 48 hr and 7, 14, 21, 28 and 35 days after the last treatment. In the hippocampus and striatum, but not in the cerebral cortex, of control rats there as a significant age-related decline of ChE activity and maximal density of 3H-QNB binding sites (Bmax). The repeated administration of DFP during the first week caused a syndrome of cholinergic stimulation both in aged and young rats. The syndrome was more pronounced, in terms of intensity and duration in aged than in young animals resulting in 40 and 12% mortality, respectively; during the second week the syndrome attenuated in the two age-groups. The percentage inhibition of brain ChE at the end of DFP treatment did not differ between young and surviving aged rats. The down-regulation of mACRs was present in the three brain regions of both young and age rats (from 20 to 40%). Factorial analysis of variance showed significant differences for age, recovery rate, and significant interaction between age and recovery rate, both for ChE and mAChRs in young rats the three brain areas

Impaired recovery of brain muscarinic receptor sites following an adaptive down-regulation induced by repeated administration of diisopropyl fluorophosphate in aged rats

Energy Technology Data Exchange (ETDEWEB)

Pintor, A.; Fortuna, S.; De Angelis, S.; Michalek, H. (Istituto Superiore di Sanita, Rome (Italy))

1990-01-01

Potential age-related differences in the recovery rate of brain cholinesterase activity (ChE) and muscarinic acetylcholine receptor binding sites (mAChRs) following reduction induced by repeated treatment with diisopropyl fluorophosphate (DFP) were evaluated in Sprague-Dawley rats. Male 3- and 24-month old rats were s.c. injected with DFP on alternate days for 2 weeks and killed 48 hr and 7, 14, 21, 28 and 35 days after the last treatment. In the hippocampus and striatum, but not in the cerebral cortex, of control rats there as a significant age-related decline of ChE activity and maximal density of 3H-QNB binding sites (Bmax). The repeated administration of DFP during the first week caused a syndrome of cholinergic stimulation both in aged and young rats. The syndrome was more pronounced, in terms of intensity and duration in aged than in young animals resulting in 40 and 12% mortality, respectively; during the second week the syndrome attenuated in the two age-groups. The percentage inhibition of brain ChE at the end of DFP treatment did not differ between young and surviving aged rats. The down-regulation of mACRs was present in the three brain regions of both young and age rats (from 20 to 40%). Factorial analysis of variance showed significant differences for age, recovery rate, and significant interaction between age and recovery rate, both for ChE and mAChRs in young rats the three brain areas.

10 CFR 72.94 - Design basis external man-induced events.

Science.gov (United States)

2010-01-01

... 10 Energy 2 2010-01-01 2010-01-01 false Design basis external man-induced events. 72.94 Section 72... WASTE Siting Evaluation Factors § 72.94 Design basis external man-induced events. (a) The region must be examined for both past and present man-made facilities and activities that might endanger the proposed...

Spatial and seasonal heterogeneity of atmospheric particles induced reactive oxygen species in urban areas and the role of water-soluble metals

International Nuclear Information System (INIS)

Gali, Nirmal Kumar; Yang, Fenhuan; Jiang, Sabrina Yanan; Chan, Ka Lok; Sun, Li; Ho, Kin-fai; Ning, Zhi

2015-01-01

Adverse health effects are associated with exposure to atmospheric particulate matter (PM), which carry various chemical constituents and induce both exogenous and endogenous oxidative stress. This study investigated the spatial and seasonal variability of PM-induced ROS at four sites with different characteristics in Hong Kong. Cytotoxicity, exogenous and endogenous ROS was determined on a dose and time dependent analysis. Large spatial variation of ROS was observed with fine PM at urban site showing highest ROS levels while coarse PM at traffic site ranks the top. No consistent seasonal difference was observed for ROS levels among all sites. The highly heterogeneous distribution of PM-induced ROS demonstrates the differential capability of PM to produce oxidative stress, and the need to use appropriate metrics as surrogates of exposure instead of PM mass in epidemiologic studies. Several transition metals were found associated with ROS by different degree illustrating the complexity of mechanisms involved. - Highlights: • Adverse health effects are associated with size segregated atmospheric PM. • Seasonal and spatial variability of PM induced ROS determined in Hong Kong city. • Coarse PM ranks top in ROS generation on per volume and mass basis. • Traffic site demonstrated as source of potent inducer of cell toxicity. • No consistent seasonal difference observed for fine and coarse PM. - Heterogeneous PM-induced ROS distribution was observed in a city. Several water-soluble metals were associated with the ROS generation but with different degree from different sites

The Earliest Chinese Proto-Porcelain Excavated from Kiln Sites: An Elemental Analysis.

Science.gov (United States)

Li, Yu; Zhang, Bin; Cheng, Huansheng; Zheng, Jianming

2015-01-01

In June 2012, the Piaoshan kiln site was excavated in Huzhou, Zhejiang Province, which hitherto proved to be the earliest known Chinese proto-porcelain kiln. Judging from the decorative patterns of unearthed impressed stoneware and proto-porcelain sherds, the site was determined to date to the late Xia (c. 2070-c. 1600 BC), the first dynasty of China. Here, we report on proton-induced X-ray emission analyses of 118 proto-porcelain and 35 impressed stoneware sherds from Piaoshan and five subsequent kiln sites in the vicinity. Using principal components analysis on the major chemical compositions, we reveal the relationships between impressed stoneware and proto-porcelain samples from the six kiln sites. The sherds from different sites have distinctive chemical profiles. The results indicate that the raw materials were procured locally. We find a developmental tendency for early glazes towards mature calcium-based glaze. It is most likely that woody plant ashes with increased calcia-potash ratios were applied to the formula.

The Earliest Chinese Proto-Porcelain Excavated from Kiln Sites: An Elemental Analysis.

Directory of Open Access Journals (Sweden)

Yu Li

Full Text Available In June 2012, the Piaoshan kiln site was excavated in Huzhou, Zhejiang Province, which hitherto proved to be the earliest known Chinese proto-porcelain kiln. Judging from the decorative patterns of unearthed impressed stoneware and proto-porcelain sherds, the site was determined to date to the late Xia (c. 2070-c. 1600 BC, the first dynasty of China. Here, we report on proton-induced X-ray emission analyses of 118 proto-porcelain and 35 impressed stoneware sherds from Piaoshan and five subsequent kiln sites in the vicinity. Using principal components analysis on the major chemical compositions, we reveal the relationships between impressed stoneware and proto-porcelain samples from the six kiln sites. The sherds from different sites have distinctive chemical profiles. The results indicate that the raw materials were procured locally. We find a developmental tendency for early glazes towards mature calcium-based glaze. It is most likely that woody plant ashes with increased calcia-potash ratios were applied to the formula.

The oxidative potential and biological effects induced by PM{sub 10} obtained in Mexico City and at a receptor site during the MILAGRO Campaign

Energy Technology Data Exchange (ETDEWEB)

Quintana, Raul [Instituto Nacional de Cancerologia, Mexico City (Mexico); Serrano, Jesus [Facultad de Ciencias, Universidad Nacional Autonoma de Mexico, Mexico City (Mexico); Gomez, Virginia [Instituto de Quimica, Universidad Nacional Autonoma de Mexico, Mexico City (Mexico); Foy, Benjamin de [Earth and Atmospheric Sciences, Saint Louis University, St. Louis, MO (United States); Miranda, Javier [Instituto de Fisica, Universidad Nacional Autonoma de Mexico, Mexico City (Mexico); Garcia-Cuellar, Claudia [Instituto Nacional de Cancerologia, Mexico City (Mexico); Vega, Elizabeth [Instituto Mexicano del Petroleo, Mexico City (Mexico); Vazquez-Lopez, Ines [Instituto Nacional de Cancerologia, Mexico City (Mexico); Molina, Luisa T. [Molina Center for Energy and the Environment, CA (United States); Massachusetts Institute of Technology, Cambridge, MA (United States); Manzano-Leon, Natalia [Instituto Nacional de Cancerologia, Mexico City (Mexico); Rosas, Irma [Centro de Ciencias de la Atmosfera, Universidad Nacional Autonoma de Mexico, Mexico City (Mexico); Osornio-Vargas, Alvaro R., E-mail: osornio@ualberta.ca [Instituto Nacional de Cancerologia, Mexico City (Mexico); Department of Paediatrics, University of Alberta, 1048 RTF, 8308 114 St, Edmonton, AB T6G 2V2 (Canada)

2011-12-15

As part of a field campaign that studied the impact of Mexico City pollution plume at the local, sub-regional and regional levels, we studied transport-related changes in PM{sub 10} composition, oxidative potential and in vitro toxicological patterns (hemolysis, DNA degradation). We collected PM{sub 10} in Mexico City (T{sub 0}) and at a suburban-receptor site (T{sub 1}), pooled according to two observed ventilation patterns (T{sub 0} {yields} T{sub 1} influence and non-influence). T{sub 0} samples contained more Cu, Zn, and carbon whereas; T{sub 1} samples contained more of Al, Si, P, S, and K (p < 0.05). Only SO{sub 4}{sup -2} increased in T{sub 1} during the influence periods. Oxidative potential correlated with Cu/Zn content (r = 0.74; p < 0.05) but not with biological effects. T{sub 1} PM{sub 10} induced greater hemolysis and T{sub 0} PM{sub 10} induced greater DNA degradation. Influence/non-influence did not affect oxidative potential nor biological effects. Results indicate that ventilation patterns had little effect on intrinsic PM{sub 10} composition and toxicological potential, which suggests a significant involvement of local sources. - Highlights: > Transport-related changes in PM{sub 10} composition, oxidative potential and in vitro toxicity were studied. > Cu, Zn, and carbon levels were predominant in urban PM{sub 10}; receptor site PM{sub 10} was rich in soil elements. > SO{sub 4}{sup -2} was the only component increased in PM{sub 10} from the receptor during the influence periods. > PM{sub 10} oxidative potential correlates with Cu/Zn content but not with studied biological effects. > Ventilation patterns had little effect on PM{sub 10} composition and toxicity. - Mexico City ventilation patterns had little effect on the intrinsic PM{sub 10} composition and toxicological potential, which suggests a significant involvement of local sources as opposed to downwind transport.

Site-specific programming of the host epithelial transcriptome by the gut microbiota

DEFF Research Database (Denmark)

Sommer, Felix; Nookaew, Intawat; Sommer, Nina

2015-01-01

BACKGROUND: The intestinal epithelium separates us from the microbiota but also interacts with it and thus affects host immune status and physiology. Previous studies investigated microbiota-induced responses in the gut using intact tissues or unfractionated epithelial cells, thereby limiting....... The microbial impact on host gene expression was highly site specific, as epithelial responses to the microbiota differed between cell fractions. Specific transcriptional regulators were enriched in each fraction. In general, the gut microbiota induced a more rapid response in the colon than in the ileum...

Inducing autophagy

DEFF Research Database (Denmark)

Harder, Lea M; Bunkenborg, Jakob; Andersen, Jens S.

2014-01-01

catabolism, which has recently been found to induce autophagy in an MTOR independent way and support cancer cell survival. In this study, quantitative phosphoproteomics was applied to investigate the initial signaling events linking ammonia to the induction of autophagy. The MTOR inhibitor rapamycin was used...... as a reference treatment to emphasize the differences between an MTOR-dependent and -independent autophagy-induction. By this means 5901 phosphosites were identified of which 626 were treatment-specific regulated and 175 were coregulated. Investigation of the ammonia-specific regulated sites supported that MTOR...

78 FR 28503 - Approval and Promulgation of Air Quality Implementation Plans; Indiana; Lake and Porter Counties...

Science.gov (United States)

2013-05-15

...: FRL-9812-4] Approval and Promulgation of Air Quality Implementation Plans; Indiana; Lake and Porter...). ACTION: Direct final rule. SUMMARY: EPA is approving Indiana's request to revise the Lake and Porter... approving new MOVES2010a-based budgets for the Lake and Porter County, Indiana 1997 8-hour ozone maintenance...

Justice according to Nsoyenyoni: An analysis of conflict resolution in ...

African Journals Online (AJOL)

Readers of the anthology AbaseGuswini LeZothamlilo may wonder if Nsoyenyoni's justice is jungle justice or something of his own making. Justice, punishment and revenge are discussed as concepts to establish their full meanings. This article also explores circumstances where Nsoyenyoni is involved in trying to level the ...

Earthquakes and associated topics in relation to nuclear power plant siting

International Nuclear Information System (INIS)

1991-01-01

This Safety Guide was prepared as part of the Agency's programme for establishing Codes and Safety Guides relating to nuclear power plants. The main purpose of the text is to provide guidance on the determination of the design basis ground motions for a nuclear power plant at a chosen site and on the determination of the potential for surface faulting at that site. Additionally, the Guide discusses other permanent displacement phenomena (liquefaction, slope instability, subsidence and collapse) and introduces the topic of seismically induced flooding. Volcanic activity is not dealt with except in connection with tsunamis. 55 refs

Induced resistance: an enhancement of basal resistance?

NARCIS (Netherlands)

Vos, M. de; Robben, C.; Pelt, J.A. van; Loon, L.C. van; Pieterse, C.M.J.

2002-01-01

Upon primary pathogen attack, plants activate resistance mechanisms at the site of infection. Besides this so-called basal resistance, plants have also the ability to enhance their defensive capacity against future pathogen attack. There are at least two types of biologically induced resistance.

Hypoxia-induced pulmonary vascular remodeling: cellular and molecular mechanisms.

Science.gov (United States)

Stenmark, Kurt R; Fagan, Karen A; Frid, Maria G

2006-09-29

Chronic hypoxic exposure induces changes in the structure of pulmonary arteries, as well as in the biochemical and functional phenotypes of each of the vascular cell types, from the hilum of the lung to the most peripheral vessels in the alveolar wall. The magnitude and the specific profile of the changes depend on the species, sex, and the developmental stage at which the exposure to hypoxia occurred. Further, hypoxia-induced changes are site specific, such that the remodeling process in the large vessels differs from that in the smallest vessels. The cellular and molecular mechanisms vary and depend on the cellular composition of vessels at particular sites along the longitudinal axis of the pulmonary vasculature, as well as on local environmental factors. Each of the resident vascular cell types (ie, endothelial, smooth muscle, adventitial fibroblast) undergo site- and time-dependent alterations in proliferation, matrix protein production, expression of growth factors, cytokines, and receptors, and each resident cell type plays a specific role in the overall remodeling response. In addition, hypoxic exposure induces an inflammatory response within the vessel wall, and the recruited circulating progenitor cells contribute significantly to the structural remodeling and persistent vasoconstriction of the pulmonary circulation. The possibility exists that the lung or lung vessels also contain resident progenitor cells that participate in the remodeling process. Thus the hypoxia-induced remodeling of the pulmonary circulation is a highly complex process where numerous interactive events must be taken into account as we search for newer, more effective therapeutic interventions. This review provides perspectives on each of the aforementioned areas.

A PCV2 vaccine based on genotype 2b is more effective than a 2a-based vaccine to protect against PCV2b or combined PCV2a/2b viremia in pigs with concurrent PCV2, PRRSV and PPV infection.

Science.gov (United States)

Opriessnig, Tanja; O'Neill, Kevin; Gerber, Priscilla F; de Castro, Alessandra M M G; Gimenéz-Lirola, Luis G; Beach, Nathan M; Zhou, Lei; Meng, Xiang-Jin; Wang, Chong; Halbur, Patrick G

2013-01-07

The predominant genotype of porcine circovirus (PCV) in the pig population today is PCV2b yet PCV2a-based commercial vaccines are considered effective in protecting against porcine circovirus associated disease. The objective of this study was to compare the ability of PCV2a- and PCV2b-based vaccines to control PCV2b viremia in a challenge model that mimics the U.S. field situation. Sixty-three pigs were randomly assigned to one of eight groups. Sixteen pigs were vaccinated with an experimental live-attenuated chimeric PCV1-2a vaccine based on genotype 2a and another 16 pigs with a chimeric PCV1-2b vaccine based on genotype 2b. Challenge was done 28 days post vaccination (dpv) using PCV2b (or a combination of PCV2a and PCV2b), porcine reproductive and respiratory syndrome virus (PRRSV), and porcine parvovirus (PPV) to mimic what commonly occurs in the field. The experiment was terminated 21 days post challenge (dpc) or 49dpv. Pigs vaccinated with the chimeric PCV1-2b vaccine had significantly higher levels of PCV1-2b viremia and shedding of the PCV1-2b vaccine virus in feces and nasal secretions but also a more robust humoral immune response as evidenced by significantly higher ELISA S/P ratios compared to the PCV1-2a vaccination. Regardless of challenge, the PCV1-2b vaccination significantly reduced the prevalence and amount of PCV2 viremia compared to the PCV1-2a vaccination. Interestingly, in the non-vaccinated pigs concurrent PCV2a infection resulted in clinical disease and increased macroscopic lung lesions compared to pigs challenged with PCV2b alone, further supporting the idea that concurrent PCV2a/PCV2b infection is necessary for optimal PCV2 replication. Copyright © 2012 Elsevier Ltd. All rights reserved.

Specificity of mutations induced by carbon ions in budding yeast Saccharomyces cerevisiae

International Nuclear Information System (INIS)

Matuo, Youichirou; Nishijima, Shigehiro; Hase, Yoshihiro; Sakamoto, Ayako; Tanaka, Atsushi; Shimizu, Kikuo

2006-01-01

To investigate the nature of mutations induced by accelerated ions in eukaryotic cells, the effects of carbon-ion irradiation were compared with those of γ-ray irradiation in the budding yeast Saccharomyces cerevisiae. The mutational effect and specificity of carbon-ion beams were studied in the URA3 gene of the yeast. Our experiments showed that the carbon ions generated more than 10 times the number of mutations induced by γ-rays, and that the types of base changes induced by carbon ions include transversions (68.7%), transitions (13.7%) and deletions/insertions (17.6%). The transversions were mainly G:C → T:A, and all the transitions were G:C → A:T. In comparison with the surrounding sequence context of mutational base sites, the C residues in the 5'-AC(A/T)-3' sequence were found to be easily changed. Large deletions and duplications were not observed, whereas ion-induced mutations in Arabidopsis thaliana were mainly short deletions and rearrangements. The remarkable feature of yeast mutations induced by carbon ions was that the mutation sites were localized near the linker regions of nucleosomes, whereas mutations induced by γ-ray irradiation were located uniformly throughout the gene

Early Site Permit Demonstration Program: Siting Guide, Site selection and evaluation criteria for an early site permit application

International Nuclear Information System (INIS)

1993-01-01

In August 1991, the Joint Contractors came to agreement with Sandia National Laboratories (SNL) and the Department of Energy (DOE) on a workscope for the cost-shared Early Site Permit Demonstration Program. One task within the scope was the development of a guide for site selection criteria and procedures. A generic Siting Guide his been prepared that is a roadmap and tool for applicants to use developing detailed siting plans for their specific region of the country. The guide presents three fundamental principles that, if used, ensure a high degree of success for an ESP applicant. First, the site selection process should take into consideration environmentally diverse site locations within a given region of interest. Second, the process should contain appropriate opportunities for input from the public. Third, the process should be applied so that it is clearly reasonable to an impartial observer, based on appropriately selected criteria, including criteria which demonstrate that the site can host an advanced light water reactor (ALWR). The Siting Guide provides for a systematic, comprehensive site selection process in which three basic types of criteria (exclusionary, avoidance, and suitability) are presented via a four-step procedure. It provides a check list of the criteria for each one of these steps. Criteria are applied qualitatively, as well as presented numerically, within the guide. The applicant should use the generic guide as an exhaustive checklist, customizing the guide to his individual situation

Current and emerging techniques for contaminant mapping and data visualization at DNAPL sites

DEFF Research Database (Denmark)

Wealthall, Gary; Durant, Neal; Grosen, Bernt

methods will be discussed in the context of contaminant delineation, remediation design, technology verification and regulatory acceptance. We present a range of site investigation tools, based on the principle of combined lines of evidence and the premise that a single technique is not available to fully......Recent advances in the development of contaminant specific site investigation tools has significantly enhanced our ability to characterize the spatial architecture of DNAPL source zones in both sedimentary and fractured bedrock environments. The application of innovative site characterization...... value. These methods are generally of lower cost, produce vertical profiles of real-time data and are primarily used to identify site areas that require further investigation. Examples of such "decision quality" methods are drive point tools, such as laser-induced fluorescence profiling and membrane...

Characteristics of the volatile organic compounds -- Arid Integrated Demonstration Site

International Nuclear Information System (INIS)

Last, G.V.; Lenhard, R.J.; Bjornstad, B.N.; Evans, J.C.; Roberson, K.R.; Spane, F.A.; Amonette, J.E.; Rockhold, M.L.

1991-10-01

The Volatile Organic Compounds -- Arid Integrated Demonstration Program (VOC-Arid ID) is targeted at demonstration and testing of technologies for the evaluation and cleanup of volatile organic compounds and associated contaminants at arid DOE sites. The initial demonstration site is an area of carbon tetrachloride (CCl 4 ) contamination located near the center of the Hanford Site. The movement of CCl 4 and other volatile organic contaminants in the subsurface is very complex. The problem at the Hanford Site is further complicated by the concurrent discharge of other waste constituents including acids, lard oil, organic phosphates, and transuranic radionuclides. In addition, the subsurface environment is very complex, with large spatial variabilities in hydraulic properties. A thorough understanding of the problem is essential to the selection of appropriate containment, retrieval, and/or in situ remedial technologies. The effectiveness of remedial technologies depends on knowing where the contaminants are, how they are held up in a given physical and chemical subsurface environment; and knowing the physical, chemical, and microbiological changes that are induced by the various remedial technologies

Processing of radiation-induced clustered DNA damage generates DSB in mammalian cells

International Nuclear Information System (INIS)

Gulston, M.K.; De Lara, C.M.; Davis, E.L.; Jenner, T.J.; O'Neill, P.

2003-01-01

Full text: Clustered DNA damage sites, in which two or more lesions are formed within a few helical turns of the DNA after passage of a single radiation track, are signatures of DNA modifications induced by ionizing radiation in mammalian cell. With 60 Co-radiation, the abundance of clustered DNA damage induced in CHO cells is ∼4x that of prompt double strand breaks (DSB) determined by PFGE. Less is known about the processing of non-DSB clustered DNA damage induced in cells. To optimize observation of any additional DSB formed during processing of DNA damage at 37 deg C, xrs-5 cells deficient in non-homologous end joining were used. Surprisingly, ∼30% of the DSB induced by irradiation at 37 deg C are rejoined within 4 minutes in both mutant and wild type cells. No significant mis-repair of these apparent DSB was observed. It is suggested that a class of non-DSB clustered DNA damage is formed which repair correctly within 4 min but, if 'trapped' prior to repair, are converted into DSB during the lysis procedure of PFGE. However at longer times, a proportion of non-DSB clustered DNA damage sites induced by γ-radiation are converted into DSB within ∼30 min following post-irradiation incubation at 37 deg C. The corresponding formation of additional DSB was not apparent in wild type CHO cells. From these observations, it is estimated that only ∼10% of the total yield of non DSB clustered DNA damage sites are converted into DSB through cellular processing. The biological consequences that the majority of non-DSB clustered DNA damage sites are not converted into DSBs may be significant even at low doses, since a finite chance exists of these clusters being formed in a cell by a single radiation track

High-throughput screening platform for engineered nanoparticle-mediated genotoxicity using CometChip technology.

Science.gov (United States)

Watson, Christa; Ge, Jing; Cohen, Joel; Pyrgiotakis, Georgios; Engelward, Bevin P; Demokritou, Philip

2014-03-25

The likelihood of intentional and unintentional engineered nanoparticle (ENP) exposure has dramatically increased due to the use of nanoenabled products. Indeed, ENPs have been incorporated in many useful products and have enhanced our way of life. However, there are many unanswered questions about the consequences of nanoparticle exposures, in particular, with regard to their potential to damage the genome and thus potentially promote cancer. In this study, we present a high-throughput screening assay based upon the recently developed CometChip technology, which enables evaluation of single-stranded DNA breaks, abasic sites, and alkali-sensitive sites in cells exposed to ENPs. The strategic microfabricated, 96-well design and automated processing improves efficiency, reduces processing time, and suppresses user bias in comparison to the standard comet assay. We evaluated the versatility of this assay by screening five industrially relevant ENP exposures (SiO2, ZnO, Fe2O3, Ag, and CeO2) on both suspension human lymphoblastoid (TK6) and adherent Chinese hamster ovary (H9T3) cell lines. MTT and CyQuant NF assays were employed to assess cellular viability and proliferation after ENP exposure. Exposure to ENPs at a dose range of 5, 10, and 20 μg/mL induced dose-dependent increases in DNA damage and cytotoxicity. Genotoxicity profiles of ZnO>Ag>Fe2O3>CeO2>SiO2 in TK6 cells at 4 h and Ag>Fe2O3>ZnO>CeO2>SiO2 in H9T3 cells at 24 h were observed. The presented CometChip platform enabled efficient and reliable measurement of ENP-mediated DNA damage, therefore demonstrating the efficacy of this powerful tool in nanogenotoxicity studies.

Clustered DNA lesion repair in eukaryotes: Relevance to mutagenesis and cell survival

Energy Technology Data Exchange (ETDEWEB)

Sage, Evelyne [Institut Curie, Bat. 110, Centre Universitaire, 91405 Orsay (France); CNRS UMR3348, Bat. 110, Centre Universitaire, 91405 Orsay (France); Harrison, Lynn, E-mail: lclary@lsuhsc.edu [Department of Molecular and Cellular Physiology, LSUHSC-S, 1501 Kings Highway, Shreveport, LA 71130 (United States)

2011-06-03

A clustered DNA lesion, also known as a multiply damaged site, is defined as {>=}2 damages in the DNA within 1-2 helical turns. Only ionizing radiation and certain chemicals introduce DNA damage in the genome in this non-random way. What is now clear is that the lethality of a damaging agent is not just related to the types of DNA lesions introduced, but also to how the damage is distributed in the DNA. Clustered DNA lesions were first hypothesized to exist in the 1990s, and work has progressed where these complex lesions have been characterized and measured in irradiated as well as in non-irradiated cells. A clustered lesion can consist of single as well as double strand breaks, base damage and abasic sites, and the damages can be situated on the same strand or opposing strands. They include tandem lesions, double strand break (DSB) clusters and non-DSB clusters, and base excision repair as well as the DSB repair pathways can be required to remove these complex lesions. Due to the plethora of oxidative damage induced by ionizing radiation, and the repair proteins involved in their removal from the DNA, it has been necessary to study how repair systems handle these lesions using synthetic DNA damage. This review focuses on the repair process and mutagenic consequences of clustered lesions in yeast and mammalian cells. By examining the studies on synthetic clustered lesions, and the effects of low vs high LET radiation on mammalian cells or tissues, it is possible to extrapolate the potential biological relevance of these clustered lesions to the killing of tumor cells by radiotherapy and chemotherapy, and to the risk of cancer in non-tumor cells, and this will be discussed.

Site Closure Strategy Model for Creosote Site

International Nuclear Information System (INIS)

Coll, F.R.; Gray, D.R.

2009-01-01

In conjunction with RCRA site corrective action at an active wood preserving facility, a risk-based site closure strategy was developed and incorporated the performance of a dense non-aqueous phase liquid (DNAPL) source recovery remedy, a monitored natural attenuation (MNA) remedy for dissolved phase groundwater, and institutional controls. Innovative creosote DNAPL source recovery has been undertaken at the Site since 1998. Pooled creosote DNAPL is present 90 feet below ground within a transmissive sand and gravel aquifer with a saturated thickness of approximately 80 feet. The creosote DNAPL source is situated on the property boundary of the site and has generated a 1/2 mile off-site dissolved phase plume, creating significant NAPL management and remedial technology verification issues. To date, over 120,000 gallons of creosote DNAPL have been recovered from the subsurface utilizing a modified circulation well technology. A mass discharge flux protocol was developed to serve as a major performance metrics for the continuation of source removal efforts and to support the application of monitored natural attenuation as an associated remedial technology for groundwater. The mass removal success has supported the MNA remedy for dissolved phase groundwater and the associated development of institutional controls. The enacted site management strategy outlines the current and future risk management activities for the Site and represents an appropriate site closure strategy for the Site. (authors)

Systemic administration of kainic acid induces selective time dependent decrease in [125I]insulin-like growth factor I, [125I]insulin-like growth factor II and [125I]insulin receptor binding sites in adult rat hippocampal formation

International Nuclear Information System (INIS)

Quirion, R.; Chabot, J.-G.; Dore, S.; Seto, D.; Kar, S.

1997-01-01

Administration of kainic acid evokes acute seizure in hippocampal pathways that results in a complex sequence of functional and structural alterations resembling human temporal lobe epilepsy. The structural alterations induced by kainic acid include selective loss of neurones in CA1-CA3 subfields and the hilar region of the dentate gyrus followed by sprouting and permanent reorganization of the synaptic connections of the mossy fibre pathways. Although the neuronal degeneration and process of reactive synaptogenesis have been extensively studied, at present little is known about means to prevent pathological conditions leading to kainate-induced cell death. In the present study, to address the role of insulin-like growth factors I and II, and insulin in neuronal survival as well as synaptic reorganization following kainate-induced seizure, the time course alterations of the corresponding receptors were evaluated. Additionally, using histological preparations, the temporal profile of neuronal degeneration and hypertrophy of resident astroglial cells were also studied. [ 125 I]Insulin-like growth factor I binding was found to be decreased transiently in almost all regions of the hippocampal formation at 12 h following treatment with kainic acid. The dentate hilar region however, exhibited protracted decreases in [ 125 I]insulin-like growth factor I receptor sites throughout (i.e. 30 days) the study. [ 125 I]Insulin-like growth factor II receptor binding sites in the hippocampal formation were found to be differentially altered following systemic administration of kainic acid. A significant decrease in [ 125 I]insulin-like growth factor II receptor sites was observed in CA1 subfield and the pyramidal cell layer of the Ammon's horn at all time points studied whereas the hilar region and the stratum radiatum did not exhibit alteration at any time. A kainate-induced decrease in [ 125 I]insulin receptor binding was noted at all time points in the molecular layer of the

Two cases of occupational allergic contact dermatitis from a cycloaliphatic epoxy resin in a neat oil

DEFF Research Database (Denmark)

Jensen, Charlotte D; Andersen, Klaus E

2003-01-01

to a neat oil used in metal processing. Patch testing revealed that the relevant contact allergen was a cycloaliphatic epoxy resin, 1,2-cyclohexanedicarboxylic acid, bis(oxiranylmethyl) ester, added to the oil as a stabilizer. None of the patients had positive reactions to the bisphenol A-based epoxy resin...... product is essential....

Site control technique for quantum dots using electron beam induced deposition

Energy Technology Data Exchange (ETDEWEB)

Iizuka, Kanji; Jung, JaeHun; Yokota, Hiroshi [Nippon Institute of Technology, 4-1 Gakuendai, Miyashiro, Minami-saitama, Saitama 3458501 (Japan)

2014-05-15

To develop simple and high throughput sit definition technique for quantum dots (QDs), the electron beam induced deposition (EBID) method was used as desorption guide of phosphorus atoms form InP substrate. As the results one or a few indium (In) droplets (DLs) were created in the carbon grid pattern by thermal annealing at a temperature of 450°C for 10 min in the ultra high vacuum condition. The size of In DLs was larger than QDs, but arsenide DLs by molecular beam in growth chamber emitted wavelength of 1.028μm at 50K by photoluminescence measurement.

Site control technique for quantum dots using electron beam induced deposition

International Nuclear Information System (INIS)

Iizuka, Kanji; Jung, JaeHun; Yokota, Hiroshi

2014-01-01

To develop simple and high throughput sit definition technique for quantum dots (QDs), the electron beam induced deposition (EBID) method was used as desorption guide of phosphorus atoms form InP substrate. As the results one or a few indium (In) droplets (DLs) were created in the carbon grid pattern by thermal annealing at a temperature of 450°C for 10 min in the ultra high vacuum condition. The size of In DLs was larger than QDs, but arsenide DLs by molecular beam in growth chamber emitted wavelength of 1.028μm at 50K by photoluminescence measurement

Combination of pentafluorophenylhydrazine derivatization and isotope dilution LC-MS/MS techniques for the quantification of apurinic/apyrimidinic sites in cellular DNA.

Science.gov (United States)

Li, Jie; Leung, Elvis M K; Choi, Martin M F; Chan, Wan

2013-05-01

Apurinic/apyrimidinic (AP) sites are common DNA lesions arising from spontaneous hydrolysis of the N-glycosidic bond and base-excision repair mechanisms of the modified bases. Due to the strong association of AP site formation with physically/chemically induced DNA damage, quantifying AP sites provides important information for risk assessment of exposure to genotoxins and oxidative stress. However, rigorous quantification of AP sites in DNA has been hampered by technical problems relating to the sensitivity and selectivity of existing analytical methods. We have developed a new isotope dilution liquid chromatography-coupled tandem mass spectrometry (LC-MS/MS) method for the rigorous quantification of AP sites in genomic DNA. The method entails enzymatic digestion of AP site-containing DNA by endo- and exonucleases, derivatization with pentafluorophenylhydrazine (PFPH), addition of an isotopically labeled PFPH derivative as internal standard, and quantification by LC-MS/MS. The combination of PFPH derivatization with LC-MS/MS analysis on a triple quadrupole mass spectrometer allows for sensitive and selective quantification of AP sites in DNA at a detection limit of 6.5 fmol, corresponding to 4 AP sites/10(9) nt in 5 μg of DNA, which is at least ten times more sensitive than existing analytical methods. The protocol was validated by AP site-containing oligonucleotides and applied in quantifying methyl methanesulfonate-induced formation of AP sites in cellular DNA.

Study of site layout in the Rokkasho site

International Nuclear Information System (INIS)

Sato, Kazuyoshi; Tamura, Kousaku; Yagenji, Akira; Sekiya, Shigeki; Takahashi, Hideo; Neyatani, Yuzuru; Uehara, Masaharu; Motohashi, Keiichi; Hashimoto, Masayoshi; Ogino, Shunji; Nagamatsu, Nobuhide

2006-03-01

The Final Design Report (FDR) of the International Thermonuclear Experimental Reactor (ITER) was published on July 2001 as a summary of the Engineering Design Activity (EDA). After the EDA, site dependent design has been investigated for the invitation of ITER toward Rokkasho Site (Iyasakadai area) in Aomori prefecture. This report describes the results of site layout of major buildings and structures of ITER in the Rokkasho-Site. The data of the ground near the site and the results of site dependent design in Japan were applied to this study. Through this study, the most appropriate site layout has been constructed with satisfaction of following conditions. (1) Bedrock level at the tokamak complex building is relatively high and it can be reduced the cost of excavation and foundation work. (2) Total amount of excavation soil for site preparation is minimized and the flexibility of the layout is ensured with flat ground level. (3) Accessibility of human and equipments, reduction of noise and vibration to the environment can be obtained. Total length of ducts and piping between buildings in site is minimized. (author)

Temporally-controlled site-specific recombination in zebrafish.

Directory of Open Access Journals (Sweden)

Stefan Hans

Full Text Available Conventional use of the site-specific recombinase Cre is a powerful technology in mouse, but almost absent in other vertebrate model organisms. In zebrafish, Cre-mediated recombination efficiency was previously very low. Here we show that using transposon-mediated transgenesis, Cre is in fact highly efficient in this organism. Furthermore, temporal control of recombination can be achieved by using the ligand-inducible CreER(T2. Site-specific recombination only occurs upon administration of the drug tamoxifen (TAM or its active metabolite, 4-hydroxy-tamoxifen (4-OHT. Cre-mediated recombination is detectable already 4 or 2 hours after administration of TAM or 4-OHT, demonstrating fast recombination kinetics. In addition, low doses of TAM allow mosaic labeling of single cells. Combined, our results show that conditional Cre/lox will be a valuable tool for both, embryonic and adult zebrafish studies. Furthermore, single copy insertion transgenesis of Cre/lox constructs suggest a strategy suitable also for other organisms.

Theory for site-site pair distribution functions of molecular fluids. II. Approximations for the Percus--Yevick site-site direct correlation functions

International Nuclear Information System (INIS)

Johnson, E.

1977-01-01

A theory for site-site pair distribution functions of molecular fluids is derived from the Ornstein-Zernike equation. Atom-atom pair distribution functions of this theory which were obtained by using different approximations for the Percus-Yevick site-site direct correlation functions are compared

Effects of site substitution and metal ion addition on doped manganites

CERN Document Server

Pradhan, A K; Roul, B K; Sahu, D R; Muralidhar, M

2002-01-01

We report transport, magnetization and transmission electron microscopy studies of the effects of A-and B-site substitution, and the addition of metal ions such as Pt, Ag and Sr, on doped ABO sub 3 perovskites, where A = La, Pr etc and B = Mn. Disorder induced by such substitution changes the behaviour of the charge-ordered (CO) state significantly. A-and B-site substitution suppresses the CO phase due to size mismatch and disorder produced by inhomogeneity. On the other hand, addition of metal ions such as Pt and Ag improves several colossal-magnetoresistance properties significantly due to microstructural effects and enhanced current percolation through grain boundaries.

A ligand peptide motif selected from a cancer patient is a receptor-interacting site within human interleukin-11.

Directory of Open Access Journals (Sweden)

Marina Cardó-Vila

Full Text Available Interleukin-11 (IL-11 is a pleiotropic cytokine approved by the FDA against chemotherapy-induced thrombocytopenia. From a combinatorial selection in a cancer patient, we isolated an IL-11-like peptide mapping to domain I of the IL-11 (sequence CGRRAGGSC. Although this motif has ligand attributes, it is not within the previously characterized interacting sites. Here we design and validate in-tandem binding assays, site-directed mutagenesis and NMR spectroscopy to show (i the peptide mimics a receptor-binding site within IL-11, (ii the binding of CGRRAGGSC to the IL-11R alpha is functionally relevant, (iii Arg4 and Ser8 are the key residues mediating the interaction, and (iv the IL-11-like motif induces cell proliferation through STAT3 activation. These structural and functional results uncover an as yet unrecognized receptor-binding site in human IL-11. Given that IL-11R alpha has been proposed as a target in human cancer, our results provide clues for the rational design of targeted drugs.

SitesIdentify: a protein functional site prediction tool

Directory of Open Access Journals (Sweden)

Doig Andrew J

2009-11-01

Full Text Available Abstract Background The rate of protein structures being deposited in the Protein Data Bank surpasses the capacity to experimentally characterise them and therefore computational methods to analyse these structures have become increasingly important. Identifying the region of the protein most likely to be involved in function is useful in order to gain information about its potential role. There are many available approaches to predict functional site, but many are not made available via a publicly-accessible application. Results Here we present a functional site prediction tool (SitesIdentify, based on combining sequence conservation information with geometry-based cleft identification, that is freely available via a web-server. We have shown that SitesIdentify compares favourably to other functional site prediction tools in a comparison of seven methods on a non-redundant set of 237 enzymes with annotated active sites. Conclusion SitesIdentify is able to produce comparable accuracy in predicting functional sites to its closest available counterpart, but in addition achieves improved accuracy for proteins with few characterised homologues. SitesIdentify is available via a webserver at http://www.manchester.ac.uk/bioinformatics/sitesidentify/

Glancing up or down: Mood management and selective social comparisons on social networking sites.

NARCIS (Netherlands)

Johnson, B.K.; Knobloch-Westerwick, S.

2014-01-01

Social networking sites (SNS) provide opportunities for mood management through selective exposure. This study tested the prediction that negative mood fosters self-enhancing social comparisons to SNS profiles. Participants were induced into positive or negative moods and then browsed manipulated

A gratuitous β-Lactamase inducer uncovers hidden active site dynamics of the Staphylococcus aureus BlaR1 sensor domain.

Science.gov (United States)

Frederick, Thomas E; Peng, Jeffrey W

2018-01-01

Increasing evidence shows that active sites of proteins have non-trivial conformational dynamics. These dynamics include active site residues sampling different local conformations that allow for multiple, and possibly novel, inhibitor binding poses. Yet, active site dynamics garner only marginal attention in most inhibitor design efforts and exert little influence on synthesis strategies. This is partly because synthesis requires a level of atomic structural detail that is frequently missing in current characterizations of conformational dynamics. In particular, while the identity of the mobile protein residues may be clear, the specific conformations they sample remain obscure. Here, we show how an appropriate choice of ligand can significantly sharpen our abilities to describe the interconverting binding poses (conformations) of protein active sites. Specifically, we show how 2-(2'-carboxyphenyl)-benzoyl-6-aminopenicillanic acid (CBAP) exposes otherwise hidden dynamics of a protein active site that binds β-lactam antibiotics. When CBAP acylates (binds) the active site serine of the β-lactam sensor domain of BlaR1 (BlaRS), it shifts the time scale of the active site dynamics to the slow exchange regime. Slow exchange enables direct characterization of inter-converting protein and bound ligand conformations using NMR methods. These methods include chemical shift analysis, 2-d exchange spectroscopy, off-resonance ROESY of the bound ligand, and reduced spectral density mapping. The active site architecture of BlaRS is shared by many β-lactamases of therapeutic interest, suggesting CBAP could expose functional motions in other β-lactam binding proteins. More broadly, CBAP highlights the utility of identifying chemical probes common to structurally homologous proteins to better expose functional motions of active sites.

Use of molecular modeling and site-directed mutagenesis to define the structural basis for the immune response to carbohydrate xenoantigens

Directory of Open Access Journals (Sweden)

Yew Margaret

2007-03-01

Full Text Available Abstract Background Natural antibodies directed at carbohydrates reject porcine xenografts. They are initially expressed in germline configuration and are encoded by a small number of structurally-related germline progenitors. The transplantation of genetically-modified pig organs prevents hyperacute rejection, but delayed graft rejection still occurs, partly due to humoral responses. IgVH genes encoding induced xenoantibodies are predominantly, not exclusively, derived from germline progenitors in the VH3 family. We have previously identified the immunoglobulin heavy chain genes encoding VH3 xenoantibodies in patients and primates. In this manuscript, we complete the structural analysis of induced xenoantibodies by identifying the IgVH genes encoding the small proportion of VH4 xenoantibodies and the germline progenitors encoding xenoantibody light chains. This information has been used to define the xenoantibody/carbohydrate binding site using computer-simulated modeling. Results The VH4-59 gene encodes antibodies in the VH4 family that are induced in human patients mounting active xenoantibody responses. The light chain of xenoantibodies is encoded by DPK5 and HSIGKV134. The structural information obtained by sequencing analysis was used to create computer-simulated models. Key contact sites for xenoantibody/carbohydrate interaction for VH3 family xenoantibodies include amino acids in sites 31, 33, 50, 57, 58 and the CDR3 region of the IgVH gene. Site-directed mutagenesis indicates that mutations in predicted contact sites alter binding to carbohydrate xenoantigens. Computer-simulated modeling suggests that the CDR3 region directly influences binding. Conclusion Xenoantibodies induced during early and delayed xenograft responses are predominantly encoded by genes in the VH3 family, with a small proportion encoded by VH4 germline progenitors. This restricted group can be identified by the unique canonical structure of the light chain, heavy

Specificity of mutations induced by carbon ions in budding yeast Saccharomyces cerevisiae

Energy Technology Data Exchange (ETDEWEB)

Matuo, Youichirou [Graduate School of Engineering, Osaka University, Yamada-oka 2-1, Suita, Osaka 565-0871 (Japan); Nishijima, Shigehiro [Graduate School of Engineering, Osaka University, Yamada-oka 2-1, Suita, Osaka 565-0871 (Japan); Hase, Yoshihiro [Radiation-Applied Biology Division, Quantum Beam Science Directorate, Japan Atomic Energy Agency (JAEA), Watanuki-machi 1233, Takasaki, Gunma 370-1292 (Japan); Sakamoto, Ayako [Radiation-Applied Biology Division, Quantum Beam Science Directorate, Japan Atomic Energy Agency (JAEA), Watanuki-machi 1233, Takasaki, Gunma 370-1292 (Japan); Tanaka, Atsushi [Radiation-Applied Biology Division, Quantum Beam Science Directorate, Japan Atomic Energy Agency (JAEA), Watanuki-machi 1233, Takasaki, Gunma 370-1292 (Japan); Shimizu, Kikuo [Radioisotope Research Center, Osaka University, Yamada-oka 2-4, Suita, Osaka 565-0871 (Japan)]. E-mail: shimizu@rirc.osaka-u.ac.jp

2006-12-01

To investigate the nature of mutations induced by accelerated ions in eukaryotic cells, the effects of carbon-ion irradiation were compared with those of {gamma}-ray irradiation in the budding yeast Saccharomyces cerevisiae. The mutational effect and specificity of carbon-ion beams were studied in the URA3 gene of the yeast. Our experiments showed that the carbon ions generated more than 10 times the number of mutations induced by {gamma}-rays, and that the types of base changes induced by carbon ions include transversions (68.7%), transitions (13.7%) and deletions/insertions (17.6%). The transversions were mainly G:C {sup {yields}} T:A, and all the transitions were G:C {sup {yields}} A:T. In comparison with the surrounding sequence context of mutational base sites, the C residues in the 5'-AC(A/T)-3' sequence were found to be easily changed. Large deletions and duplications were not observed, whereas ion-induced mutations in Arabidopsis thaliana were mainly short deletions and rearrangements. The remarkable feature of yeast mutations induced by carbon ions was that the mutation sites were localized near the linker regions of nucleosomes, whereas mutations induced by {gamma}-ray irradiation were located uniformly throughout the gene.

On-site inspection: A brief overview and bibliography of techniques pertinent to assessing suspected nuclear test sites

International Nuclear Information System (INIS)

Carrigan, C.R.

1993-03-01

The purpose of this report is to provide a brief overview and bibliography of those techniques that may have application for the evaluation of a site to determine if a high energy release event is nuclear in nature. This effort is motivated by recognition of the changing world political climate and the perception that low yield and non-proliferation issues will grow in importance as countries become increasingly involved as signators to treaties that are intended to limit the development and testing of nuclear weapons. Along with an increasing interest in such issues is the awareness of the need to implement improved capabilities for treaty monitoring programs that must deal with assessing suspicious occurrences of high energy release events. In preparing this report, it is recognized that monitoring can take two main forms. The first involves the resolution of unidentified events detected by seismic and satellite National Technical Means. Events of an indeterminate nature could occur world-wide and could induce tension in neighboring countries. If an on-site measurement capability were available, a monitoring team could be sent to the suspected site of an event to take measurements that could confirm or disprove the occurrence of a clandestine nuclear test. The second monitoring form is the confirmation that a clandestine event is not masked by a declared event. For example, a large mining explosion could mask a decoupled nuclear explosion. On-site measurements before and during the test could confirm that a clandestine event did not occur and could provide assurance that the party carrying out the explosion is not taking advantage of clandestine testing opportunities. 48 refs

Formation of radiation induced chromosome aberrations: involvement of telomeric sequences and telomerase

International Nuclear Information System (INIS)

Pirzio, L.

2004-07-01

As telomeres are crucial for chromosome integrity; we investigated the role played by telomeric sequences in the formation and in the transmission of radio-induced chromosome rearrangements in human cells. Starting from interstitial telomeric sequences (ITS) as putative region of breakage, we showed that the radiation sensitivity is not equally distributed along chromosomes and. is not affected by ITS. On the contrary, plasmid integration sites are prone to radio-induced breaks, suggesting a possible integration at sites already characterized by fragility. However plasmids do not preferentially insert at radio-induced breaks in human cells immortalized by telomerase. These cells showed remarkable karyotype stability even after irradiation, suggesting a role of telomerase in the genome maintenance despite functional telomeres. Finally, we showed that the presence of more breaks in a cell favors the repair, leading to an increase of transmissible rearrangements. (author)

Dietary flavonoid kaempferol inhibits glucocorticoid-induced bone loss by promoting osteoblast survival.

Science.gov (United States)

Adhikary, Sulekha; Choudhary, Dharmendra; Ahmad, Naseer; Karvande, Anirudha; Kumar, Avinash; Banala, Venkatesh Teja; Mishra, Prabhat Ranjan; Trivedi, Ritu

2018-02-13

Kaempferol, a dietary flavonoid found in fruits and vegetables, has been reported to reverse osteopenic condition in ovariectomized rats. Because kaempferol is endowed with osteogenic activity, the aim of this study was to determine whether it has a beneficial effect on glucocorticoid (GC)-induced bone loss. Adult female rats were divided into four groups as control (vehicle; distilled water), methylprednisolone (MP; 5 mg•kg•d, subcutaneously), MP + kaempferol (5 mg•kg•d, oral), and MP + human parathyroid 1-34 (30 µg/kg, 5 times/wk, subcutaneously) and treated for 4 wk. To study the antagonizing effect of kaempferol on GC-induced inhibition of fracture healing, drill-hole injury was performed on control and GC-treated rats. An oral dose of kaempferol was given for 14 d to observe the effect on callus formation at the site of injury. After treatment, bones were collected for further analysis. GC was associated with a decreased bone mineral density and impaired bone microarchitecture parameters. Consumption of kaempferol induced bone-sparing effects in GC-induced osteopenic condition. Additionally, improved callus formation at site of drill injury in femur diaphysis was observed with kaempferol consumption in animals on GC. Consistent with the in vivo data, kaempferol elicited a higher expression of osteogenic markers in vitro and antagonized the apoptotic effect of dexamethasone on calvarial osteoblasts. These results suggested that kaempferol reduced GC-induced bone loss and enhanced bone regeneration at fractured site, thus emphasizing the positive role of flavonoids on bone health. Copyright © 2018 Elsevier Inc. All rights reserved.

Effect of melatonin on methamphetamine- and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced dopaminergic neurotoxicity and methamphetamine-induced behavioral sensitization.

Science.gov (United States)

Itzhak, Y; Martin, J L; Black, M D; Ali, S F

1998-06-01

Methamphetamine (METH)- and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced dopaminergic neurotoxicity is thought to be associated with the formation of free radicals. Since evidence suggests that melatonin may act as a free radical scavenger and antioxidant, the present study was undertaken to investigate the effect of melatonin on METH- and MPTP-induced neurotoxicity. In addition, the effect of melatonin on METH-induced locomotor sensitization was investigated. The administration of METH (5 mg kg(-1) x 3) or MPTP (20 mg kg(-1) x 3) to Swiss Webster mice resulted in 45-57% depletion in the content of striatal dopamine and its metabolites, 3,4-dihydroxyphenylacetic acid and homovanillic acid, and 57-59% depletion in dopamine transporter binding sites. The administration of melatonin (10 mg kg(-1)) before each of the three injections of the neurotoxic agents (on day 1), and thereafter for two additional days, afforded a full protection against METH-induced depletion of dopamine and its metabolites and dopamine transporter binding sites. In addition, melatonin significantly diminished METH-induced hyperthermia. However, the treatment with melatonin had no significant effect on MPTP-induced depletion of the dopaminergic markers tested. In the set of behavioral experiments, we found that the administration of 1 mg kg(-1) METH to Swiss Webster mice for 5 days resulted in marked locomotor sensitization to a subsequent challenge injection of METH, as well as context-dependent sensitization (conditioning). The pretreatment with melatonin (10 mg kg(-1)) prevented neither the sensitized response to METH nor the development of conditioned locomotion. Results of the present study indicate that melatonin has a differential effect on the dopaminergic neurotoxicity produced by METH and MPTP. Since it is postulated that METH-induced hyperthermia is related to its neurotoxic effect, while regulation of body temperature is unrelated to MPTP-induced neurotoxicity or METH-induced

Photoacoustic imaging of angiogenesis in subdermal islet transplant sites

Science.gov (United States)

Shi, Wei; Pawlick, Rena; Bruni, Antonio; Rafiei, Yasmin; Pepper, Andrew R.; Gala-Lopez, Boris; Choi, Min; Malcolm, Andrew; Zemp, Roger J.; Shapiro, A. M. James

2016-03-01

Exogenous insulin administration is the mainstay treatment therapy for patients with Type-1 diabetes mellitus (T1DM). However, for select patients, clinical islet transplantation is an alternative therapeutic treatment. In this procedure, islets are transplanted into the hepatic portal vein, and despite improved success within the last decade, obstacles are still associated with this approach. It has been discovered that the subcutaneous space may be an effective alternative site for islet transplantation, and may provide advantages of easy access and potential for simple monitoring. The ability to monitor islet viability and the transplant microenvironment may be key to future success in islet transplantation. A subcutaneous device-less technique has been developed to facilitate angiogenesis in the islet transplant site, however, a method for monitoring the potential engraftment site have yet to be explored fully. Here we demonstrate the ability to track angiogenesis in mice with 1, 2, 3 and 4 weeks post-catheter implant on both sides of the abdomen using a FujiFilm VisualSonics Vevo-LAZR system. Quantitative analysis on vessel densities exhibited gradual vessel growth successfully induced by catheter implantation. Our study demonstrates the ability of employing photoacoustic and micro-ultrasound imaging to track angiogenesis around the catheter site prior to islet transplantation.

Quantum pumping induced by disorder in one dimension

Energy Technology Data Exchange (ETDEWEB)

Qin, Jihong [Department of Physics, University of Science and Technology Beijing, Beijing 100083 (China); Guo, Huaiming, E-mail: hmguo@buaa.edu.cn [Department of Physics, Beihang University, Beijing 100191 (China)

2016-07-01

The topological property in one dimension is protected by symmetry. Based on a concrete model, we study the effect of disorder preserving or breaking the symmetry and show the nature of symmetry protecting in the one dimensional topological phase. A stable quantum pumping can be constructed within the topological model. It is shown that an integer charge is pumped across a periodic chain in a cyclic process. Furthermore we find that not only the quantum pumping is stable to on-site disorder, but also can be induced by it. These results may be realized experimentally using quasicrystals. - Highlights: • We study the effect of disorder preserving or breaking the symmetry. • We show that an integer charge is pumped across a periodic chain in a cyclic process. • Not only the quantum pumping is stable to on-site disorder, but also can be induced by it.

Localisation of aphidicolin-induced break points in Holstein-Friesian cattle (Bos taurus using RBG-banding

Directory of Open Access Journals (Sweden)

Mernies Beatriz

2002-11-01

Full Text Available Abstract Fragile sites (FS seem to play a role in genome instability and may be involved in karyotype evolution and chromosome aberrations. The majority of common fragile sites are induced by aphidicolin. Aphidicolin was used at two different concentrations (0.15 and 0.30 μM to study the occurrence of FS in the cattle karyotype. In this paper, a map of aphidicolin induced break points and fragile sites in cattle chromosomes was constructed. The statistical analysis indicated that any band with three or more breaks was significantly damaged (P r = 0.54. On the contrary, 21 FS were identified on negative R bands while 9 FS were located on positive R bands.

Integrin activation dynamics between the RGD-binding site and the headpiece hinge.

Science.gov (United States)

Puklin-Faucher, Eileen; Vogel, Viola

2009-12-25

Integrins form mechanical links between the extracellular matrix and the cytoskeleton. Although integrin activation is known to be regulated by an allosteric conformational change, which can be induced from the extracellular or intracellular end of the molecule, little is known regarding the sequence of structural events by which signals propagate between distant sites. Here, we reveal with molecular dynamics simulations of the FnIII(10)-bound alpha(V)beta(3) integrin headpiece how the binding pocket and interdomain betaA/hybrid domain hinge on the distal end of the betaA domain are allosterically linked via a hydrophobic T-junction between the middle of the alpha1 helix and top of the alpha7 helix. The key results of this study are: 1) that this T-junction is induced by ligand binding and hinge opening, and thus displays bidirectionality; 2) that formation of this junction can be accelerated by ligand-mediated force; and 3) how formation of this junction is inhibited by Ca(2+) in place of Mg(2+) at the site adjacent to the metal ion-dependent adhesion site ("ADMIDAS"). Together with recent experimental evidence that integrin complexes can form catch bonds (i.e. become strengthened under force), as well as earlier evidence that Ca(2+) at the ADMIDAS results in lower binding affinity, these simulations provide a common structural model for the dynamic process by which integrins become activated.

Early Site Permit Demonstration Program: Siting Guide, Site selection and evaluation criteria for an early site permit application. Revision 1

Energy Technology Data Exchange (ETDEWEB)

1993-03-24

In August 1991, the Joint Contractors came to agreement with Sandia National Laboratories (SNL) and the Department of Energy (DOE) on a workscope for the cost-shared Early Site Permit Demonstration Program. One task within the scope was the development of a guide for site selection criteria and procedures. A generic Siting Guide his been prepared that is a roadmap and tool for applicants to use developing detailed siting plans for their specific region of the country. The guide presents three fundamental principles that, if used, ensure a high degree of success for an ESP applicant. First, the site selection process should take into consideration environmentally diverse site locations within a given region of interest. Second, the process should contain appropriate opportunities for input from the public. Third, the process should be applied so that it is clearly reasonable to an impartial observer, based on appropriately selected criteria, including criteria which demonstrate that the site can host an advanced light water reactor (ALWR). The Siting Guide provides for a systematic, comprehensive site selection process in which three basic types of criteria (exclusionary, avoidance, and suitability) are presented via a four-step procedure. It provides a check list of the criteria for each one of these steps. Criteria are applied qualitatively, as well as presented numerically, within the guide. The applicant should use the generic guide as an exhaustive checklist, customizing the guide to his individual situation.

Preliminary siting characterization Salt Disposition Facility - Site B

International Nuclear Information System (INIS)

Wyatt, D.

2000-01-01

A siting and reconnaissance geotechnical program has been completed in S-Area at the Savannah River Site in South Carolina. This program investigated the subsurface conditions for the area known as ''Salt Disposition Facility (SDF), Site B'' located northeast of H-Area and within the S-Area. Data acquired from the Site B investigation includes both field exploration and laboratory test data

Testicular regulation of neuronal glucose and monocarboxylate transporter gene expression profiles in CNS metabolic sensing sites during acute and recurrent insulin-induced hypoglycemia.

Science.gov (United States)

Vavaiya, Kamlesh V; Paranjape, Sachin A; Briski, Karen P

2007-01-01

Recurrent insulin-induced hypoglycemia (RIIH) impairs glucose counter-regulatory function in male humans and rodents and, in the latter, diminishes neuronal activation in CNS structures that monitor metabolic homeostasis, including the lateral hypothalamic area (LHA) and dorsal vagal complex (DVC). We investigated whether habituated neuronal reactivity in CNS sensing sites to hypoglycemia is correlated with modified monocarboxylate and/or glucose uptake by using quantitative real-time RT-PCR to analyze neuronal monocarboxylate transporter (MCT2) and glucose transporter variant (GLUT and GLUT4) gene expression profiles in the microdissected LHA, ventromedial nucleus hypothalamus (VMH), and DVC after one or multiple insulin injections. Because orchidectomy (ORDX) maintains uniform glycemic responses to RIIH in male rats, we also examined whether regional gene response patterns are testes dependent. In the intact male rat DVC, MCT2, GLUT3, and GLUT4 gene expression was not altered by acute hypoglycemia but was enhanced by RIIH. MCT2 and GLUT3 mRNA levels in the ORDX rat DVC did not differ among groups, but GLUT4 transcripts were progressively increased by acute and recurrent hypoglycemia. Precedent hypoglycemia decreased or increased basal MCT2 and GLUT4 gene expression, respectively, in the intact rat LHA; LHA GLUT3 transcription was augmented by RIIH in intact rats only. Acute hypoglycemia suppressed MCT2, GLUT3, and GLUT4 gene expression in the intact rat VMH, a response that was abolished by RIIH. In ORDX rats, VMH gene transcript levels were unchanged in response to one dose of insulin but were selectively diminished during RIIH. These data demonstrate site-specific, testes-dependent effects of acute and recurrent hypoglycemia on neuronal metabolic substrate transporter gene expression in characterized rat brain metabolic sensing loci and emphasize the need to assess the impact of potential alterations in glucose and lactate uptake during RIIH on general and

Automatic web site authoring with SiteGuide

NARCIS (Netherlands)

de Boer, V.; Hollink, V.; van Someren, M.W.; Kłopotek, M.A.; Przepiórkowski, A.; Wierzchoń, S.T.; Trojanowski, K.

2009-01-01

An important step in the design process for a web site is to determine which information is to be included and how the information should be organized on the web site’s pages. In this paper we describe ’SiteGuide’, a tool that automatically produces an information architecture for a web site that a

The allosteric site regulates the voltage sensitivity of muscarinic receptors.

Science.gov (United States)

Hoppe, Anika; Marti-Solano, Maria; Drabek, Matthäus; Bünemann, Moritz; Kolb, Peter; Rinne, Andreas

2018-01-01

Muscarinic receptors (M-Rs) for acetylcholine (ACh) belong to the class A of G protein-coupled receptors. M-Rs are activated by orthosteric agonists that bind to a specific site buried in the M-R transmembrane helix bundle. In the active conformation, receptor function can be modulated either by allosteric modulators, which bind to the extracellular receptor surface or by the membrane potential via an unknown mechanism. Here, we compared the modulation of M 1 -Rs and M 3 -Rs induced by changes in voltage to their allosteric modulation by chemical compounds. We quantified changes in receptor signaling in single HEK 293 cells with a FRET biosensor for the G q protein cycle. In the presence of ACh, M 1 -R signaling was potentiated by voltage, similarly to positive allosteric modulation by benzyl quinolone carboxylic acid. Conversely, signaling of M 3 -R was attenuated by voltage or the negative allosteric modulator gallamine. Because the orthosteric site is highly conserved among M-Rs, but allosteric sites vary, we constructed "allosteric site" M 3 /M 1 -R chimeras and analyzed their voltage dependencies. Exchanging the entire allosteric sites eliminated the voltage sensitivity of ACh responses for both receptors, but did not affect their modulation by allosteric compounds. Furthermore, a point mutation in M 3 -Rs caused functional uncoupling of the allosteric and orthosteric sites and abolished voltage dependence. Molecular dynamics simulations of the receptor variants indicated a subtype-specific crosstalk between both sites, involving the conserved tyrosine lid structure of the orthosteric site. This molecular crosstalk leads to receptor subtype-specific voltage effects. Copyright © 2017 Elsevier Inc. All rights reserved.

Conserved structural chemistry for incision activity in structurally non-homologous apurinic/apyrimidinic endonuclease APE1 and endonuclease IV DNA repair enzymes.

Energy Technology Data Exchange (ETDEWEB)

Tsutakawa, Susan E.; Shin, David S.; Mol, Clifford D.; Izum, Tadahide; Arvai, Andrew S.; Mantha, Anil K.; Szczesny, Bartosz; Ivanov, Ivaylo N.; Hosfield, David J.; Maiti, Buddhadev; Pique, Mike E.; Frankel, Kenneth A.; Hitomi, Kenichi; Cunningham, Richard P.; Mitra, Sankar; Tainer, John A.

2013-03-22

Non-coding apurinic/apyrimidinic (AP) sites in DNA form spontaneously and as DNA base excision repair intermediates are the most common toxic and mutagenic in vivo DNA lesion. For repair, AP sites must be processed by 5' AP endonucleases in initial stages of base repair. Human APE1 and bacterial Nfo represent the two conserved 5' AP endonuclease families in the biosphere; they both recognize AP sites and incise the phosphodiester backbone 5' to the lesion, yet they lack similar structures and metal ion requirements. Here, we determined and analyzed crystal structures of a 2.4 ? resolution APE1-DNA product complex with Mg(2+) and a 0.92 Nfo with three metal ions. Structural and biochemical comparisons of these two evolutionarily distinct enzymes characterize key APE1 catalytic residues that are potentially functionally similar to Nfo active site components, as further tested and supported by computational analyses. We observe a magnesium-water cluster in the APE1 active site, with only Glu-96 forming the direct protein coordination to the Mg(2+). Despite differences in structure and metal requirements of APE1 and Nfo, comparison of their active site structures surprisingly reveals strong geometric conservation of the catalytic reaction, with APE1 catalytic side chains positioned analogously to Nfo metal positions, suggesting surprising functional equivalence between Nfo metal ions and APE1 residues. The finding that APE1 residues are positioned to substitute for Nfo metal ions is supported by the impact of mutations on activity. Collectively, the results illuminate the activities of residues, metal ions, and active site features for abasic site endonucleases.

Changes in the surface electronic states of semiconductor fine particles induced by high energy ion irradiation

Energy Technology Data Exchange (ETDEWEB)

Yamaki, Tetsuya; Asai, Keisuke; Ishigure, Kenkichi [Tokyo Univ. (Japan); Shibata, Hiromi

1997-03-01

The changes in the surface electronic states of Q-sized semiconductor particles in Langmuir-Blodgett (LB) films, induced by high energy ion irradiation, were examined by observation of ion induced emission and photoluminescence (PL). Various emission bands attributed to different defect sites in the band gap were observed at the initial irradiation stage. As the dose increased, the emissions via the trapping sites decreased in intensity while the band-edge emission developed. This suggests that the ion irradiation would remove almost all the trapping sites in the band gap. The low energy emissions, which show a multiexponential decay, were due to a donor-acceptor recombination between the deeply trapped carriers. It was found that the processes of formation, reaction, and stabilization of the trapping sites would predominantly occur under the photooxidizing conditions. (author)

An approach to rehabilitation objectives. Example of hydrocarbon polluted sites; Approche des objectifs de rehabilitation. Exemple des sites pollues par des hydrocarbures

Energy Technology Data Exchange (ETDEWEB)

Chevallier, B [Union Francaise des Industries Petrolieres (UFIP), 75 - Paris (France)

1997-12-31

The UFIP (French Petroleum Industry Association) studies concerning polluted sites and soils and their rehabilitation are presented: characterization of the admissible maximum fuel or gas oil content in the ground, the issue of time effects, and the risks it may induce for public health; study on the various pollutant impacts on the environment (underground waters, biotope...); approach to the development of a dynamical model describing these phenomena and their influence on man, in order to derive consistent and reasonable decontamination objectives

An approach to rehabilitation objectives. Example of hydrocarbon polluted sites; Approche des objectifs de rehabilitation. Exemple des sites pollues par des hydrocarbures

Energy Technology Data Exchange (ETDEWEB)

Chevallier, B. [Union Francaise des Industries Petrolieres (UFIP), 75 - Paris (France)

1996-12-31

The UFIP (French Petroleum Industry Association) studies concerning polluted sites and soils and their rehabilitation are presented: characterization of the admissible maximum fuel or gas oil content in the ground, the issue of time effects, and the risks it may induce for public health; study on the various pollutant impacts on the environment (underground waters, biotope...); approach to the development of a dynamical model describing these phenomena and their influence on man, in order to derive consistent and reasonable decontamination objectives

The value of DCIP geophysical surveys for contaminated site investigations

DEFF Research Database (Denmark)

Balbarini, Nicola; Rønde, Vinni Kampman; Maurya, Pradip Kumar

an old factory site by combining traditional geological, hydrological, and contaminant concentration data with DCIP surveys. The plume consisted of xenobiotic organic compounds and inorganics. The study assesses benefits and limitations of DCIP geophysics for contaminated site investigations. A 3D......Geophysical methods are increasingly being used in contaminant hydrogeology to map lithology, hydraulic properties, and contaminant plumes with a high ionic strength. Advances in the Direct Current resistivity and Induced Polarization (DCIP) method allow the collection of high resolution three...... water and below the streambed. Surface DCIP surveys supported the characterization of the spatial variability in geology, hydraulic conductivity and contaminant concentration. Though DCIP data interpretation required additional borehole data, the DCIP survey reduced the number of boreholes required...

Hypoxia-inducible factor-1α induces multidrug resistance protein in colon cancer

Directory of Open Access Journals (Sweden)

Lv Y

2015-07-01

Full Text Available Yingqian Lv, Shan Zhao, Jinzhu Han, Likang Zheng, Zixin Yang, Li Zhao Department of Oncology, The Second Hospital, Hebei Medical University, Shijiazhuang, Hebei Province, People’s Republic of China Abstract: Multidrug resistance is the major cause of chemotherapy failure in many solid tumors, including colon cancer. Hypoxic environment is a feature for all solid tumors and is important for the development of tumor resistance to chemotherapy. Hypoxia-inducible factor (HIF-1α is the key transcription factor that mediates cellular response to hypoxia. HIF-1α has been shown to play an important role in tumor resistance; however, the mechanism is still not fully understood. Here, we found that HIF-1α and the drug resistance-associated gene multidrug resistance associated protein 1 (MRP1 were induced by treatment of colon cancer cells with the hypoxia-mimetic agent cobalt chloride. Inhibition of HIF-1α by RNA interference and dominant-negative protein can significantly reduce the induction of MRP1 by hypoxia. Bioinformatics analysis showed that a hypoxia response element is located at -378 to -373 bp upstream of the transcription start site of MRP1 gene. Luciferase reporter assay combined with mutation analysis confirmed that this element is essential for hypoxia-mediated activation of MRP gene. Furthermore, RNA interference revealed that HIF-1α is necessary for this hypoxia-driven activation of MRP1 promoter. Importantly, chromatin immunoprecipitation analysis demonstrated that HIF-1α could directly bind to this HRE site in vivo. Together, these data suggest that MRP1 is a downstream target gene of HIF-1α, which provides a potential novel mechanism for HIF-1α-mediated drug resistance in colon cancer and maybe other solid tumors as well. Keywords: hypoxia, hypoxia-inducible factor-1α, multidrug resistance associated protein, transcriptional regulation, chemotherapy tolerance

The Analysis of the Relationship between Well Being and the Levels of Emotional Abuse of the Teacher Candidates Perceive

Science.gov (United States)

Sakar, Zuleyha; Akca, Figen; Bozkurt, Aysegul

2017-01-01

Emotional abuse can be defined as a large behavior pattern which the individual is exposed to and can be compiled as, the abasement, violent attitude, exclusion, continuous criticism, unfulfilled need for love and making someone feel worthless. From this point of view the effect on the well being is a key concern about the individuals exposed to…

Two Years of Site Diversity Measurements in Guam, USA

Science.gov (United States)

Acosta, Roberto J.; Morse, J.; Zemba, M.; Nessel, J.

2012-01-01

As NASA communication networks upgrade to higher frequencies, such as Ka-Band, atmospherically induced attenuation can become significant. This attenuation is caused by rain, clouds and atmospheric gases (oxygen and water vapor), with rain having the most noticeable effects. One technique to circumvent the increase in attenuation is to operate two terminals separated by a distance that exceeds the average rain cell size. The fact that rain cells are of finite size can then be exploited by rerouting the signal to the terminal with the strongest link. This technique, known as site diversity, is best suited for climates that have compact (less than 2km) and intense rain cells such as in Guam. In order to study the potential diversity gain at the Tracking and Data Relay Satellite (TDRS) Remote Ground Terminal (GRGT) complex in Guam a site test interferometer (STI) was installed in May of 2010. The STI is composed of two terminals with a 900m baseline that observe the same unmodulated beacon signal broadcast from a geostationary satellite (e.g., UFO 8). The potential site diversity gain is calculated by measuring the difference in signal attenuation seen at each terminal. Over the two years of data collection the cumulative distribution function (CDF) of the site diversity gain shows a better than 3 dB improvement for 90% of the time over standard operation. These results show that the use of site diversity in Guam can be very effective in combating rain fades.

XRCC1 suppresses somatic hypermutation and promotes alternative nonhomologous end joining in Igh genes.

Science.gov (United States)

Saribasak, Huseyin; Maul, Robert W; Cao, Zheng; McClure, Rhonda L; Yang, William; McNeill, Daniel R; Wilson, David M; Gearhart, Patricia J

2011-10-24

Activation-induced deaminase (AID) deaminates cytosine to uracil in immunoglobulin genes. Uracils in DNA can be recognized by uracil DNA glycosylase and abasic endonuclease to produce single-strand breaks. The breaks are repaired either faithfully by DNA base excision repair (BER) or mutagenically to produce somatic hypermutation (SHM) and class switch recombination (CSR). To unravel the interplay between repair and mutagenesis, we decreased the level of x-ray cross-complementing 1 (XRCC1), a scaffold protein involved in BER. Mice heterozygous for XRCC1 showed a significant increase in the frequencies of SHM in Igh variable regions in Peyer's patch cells, and of double-strand breaks in the switch regions during CSR. Although the frequency of CSR was normal in Xrcc1(+/-) splenic B cells, the length of microhomology at the switch junctions decreased, suggesting that XRCC1 also participates in alternative nonhomologous end joining. Furthermore, Xrcc1(+/-) B cells had reduced Igh/c-myc translocations during CSR, supporting a role for XRCC1 in microhomology-mediated joining. Our results imply that AID-induced single-strand breaks in Igh variable and switch regions become substrates simultaneously for BER and mutagenesis pathways.

Plastid ribosome pausing is induced by multiple features and is linked to protein complex assembly

DEFF Research Database (Denmark)

Gawroński, Piotr; Jensen, Poul Erik; Karpinski, Stanislaw

2018-01-01

Many mRNAs contain pause sites that briefly interrupt the progress of translation. Specific features that induce ribosome pausing have been described; however, their individual contributions to pause-site formation, and the overall biological significance of ribosome pausing, remain largely uncle...

Aluminium allergy and granulomas induced by vaccinations for children

DEFF Research Database (Denmark)

Andersen, Rosa Marie O; Zachariae, Claus; Johansen, Jeanne Duus

2014-01-01

Vaccination with aluminium-adsorbed vaccines can induce aluminium allergy with persistent itching subcutaneous nodules at the injection site - vaccination granulomas. In this article we give an overview of childhood aluminium-adsorbed vaccines available in Denmark. Through literature studies we...

A novel subcutaneous site of islet transplantation superior to the liver.

Science.gov (United States)

Yasunami, Yohichi; Nakafusa, Yuki; Nitta, Naoyoshi; Nakamura, Masafumi; Goto, Masafumi; Ono, Junko; Taniguchi, Masaru

2018-03-08

Islet transplantation is an attractive treatment for patients with insulin-dependent diabetes mellitus, and currently the liver is the favored transplantation site. However, an alternative site is desirable because of the low efficiency of hepatic transplantation, requiring 2-3 donors for a single recipient, and because the transplanted islets cannot be accessed or retrieved. We developed a novel procedure of islet transplantation to the inguinal subcutaneous white adipose tissue (ISWAT) of mice and described functional and morphological characteristics of transplanted syngeneic islets. Also, it was determined whether islet allograft rejection in the ISWAT can be prevented by immunosuppressive agents. Furthermore, it was examined whether human islets function when grafted in this particular site of immune-deficient mice. In this site, transplanted islets are engrafted as clusters and function to reverse STZ-induced diabetes in mice. Importantly, transplanted islets can be visualized by CT and are easily retrievable, and allograft rejection is preventable by blockade of co-stimulatory signals. Of much importance, the efficiency of islet transplantation in this site is superior to the liver, in which hyperglycemia of diabetic recipient mice is ameliorated after transplantation of 200 syngeneic islets (the islet number yielded from 1 mouse pancreas) to the ISWAT but not to the liver. Furthermore, human islets transplanted in this particular site function to reverse diabetes in immune-deficient mice. Thus, the ISWAT is superior to the liver as the site of islet transplantation, which may lead to improved outcome of clinical islet transplantation.

Site description of Forsmark at completion of the site investigation phase. SDM-Site Forsmark

Energy Technology Data Exchange (ETDEWEB)

2008-12-15

The Swedish Nuclear Fuel and Waste Management Co., SKB, has undertaken site characterisation in two different areas, Forsmark and Laxemar-Simpevarp, in order to identify a suitable location for a geological repository of spent nuclear fuel according to the KBS-3 method. The site investigations have been conducted in campaigns, punctuated by data freezes. After each data freeze, the site data have been analysed and modelling has been carried out with the overall purpose to develop a site descriptive model (SDM). The site descriptive model is used by repository engineering to design the underground facility and to develop a repository layout adapted to the site. It is also essential for safety assessment, since the model is the only source for site-specific input. Another important use of the site descriptive model is in the environmental impact assessment. An SDM is an integrated model for geology, thermal properties, rock mechanics, hydrogeology, hydrogeochemistry, bedrock transport properties and a description of the surface system. The site descriptive model compiled in the current report, SDM-Site, presents an integrated understanding of the Forsmark area at the completion of the surface-based investigations, which were conducted at Forsmark during the period 2002 to 2007. It also provides a summary of the abundant underlying data and the discipline-specific models that support the site understanding. The description relies heavily on background reports that address, in particular, details in data analyses and modelling in the different disciplines. The Forsmark area is located in northern Uppland within the municipality of Oesthammar, about 120 km north of Stockholm. The candidate area for site investigation is located along the shoreline of Oeregrundsgrepen, within the north-western part of a major tectonic lens that formed between 1.87 and 1.85 billion years ago during the Svecokarelian orogeny. The candidate area is approximately 6 km long and 2 km wide. The

Site description of Laxemar at completion of the site investigation phase. SDM-Site Laxemar

Energy Technology Data Exchange (ETDEWEB)

2009-12-15

The Swedish Nuclear Fuel and Waste Management Company (SKB) has undertaken site characterisation in two different areas, Forsmark and Laxemar-Simpevarp, in order to identify a suitable location for a geological repository of spent nuclear fuel according to the KBS-3 method. The site investigations have been conducted in campaigns, punctuated by data freezes. After each data freeze, the site data have been analysed and modelling has been carried out with the overall purpose to develop a site descriptive model (SDM). The site descriptive model is used by repository engineering to design the underground facility and to develop a repository layout adapted to the site. It is also essential for safety assessment, since the SDM is the only source for site-specific input. Another important use of the site descriptive model is in the environmental impact assessment. An SDM is an integrated model of geology, thermal properties, rock mechanics, hydrogeology, hydrogeochemistry, bedrock transport properties and a description of the surface system. The site descriptive model compiled in the current report, SDM-Site Laxemar, presents an integrated understanding of the Laxemar-Simpevarp area (with special emphasis on the Laxemar subarea) at the completion of the surface-based investigations, which were conducted during the period 2002 to 2007. A summary is also provided of the abundant underlying data and the discipline specific models that support the site understanding. The description relies heavily on background reports that address, in particular, details of the data analyses and modelling of the different disciplines. The Laxemar-Simpevarp area is located in the province of Smaaland within the municipality of Oskarshamn, about 230 km south of Stockholm. The candidate area for site investigation is located along the shoreline of the strait of Kalmarsund, within a 1.8 billion year old suite of well preserved bedrock belonging to the Transscandinavian Igneous Belt formed during

Site description of Laxemar at completion of the site investigation phase. SDM-Site Laxemar

International Nuclear Information System (INIS)

2009-12-01

The Swedish Nuclear Fuel and Waste Management Company (SKB) has undertaken site characterisation in two different areas, Forsmark and Laxemar-Simpevarp, in order to identify a suitable location for a geological repository of spent nuclear fuel according to the KBS-3 method. The site investigations have been conducted in campaigns, punctuated by data freezes. After each data freeze, the site data have been analysed and modelling has been carried out with the overall purpose to develop a site descriptive model (SDM). The site descriptive model is used by repository engineering to design the underground facility and to develop a repository layout adapted to the site. It is also essential for safety assessment, since the SDM is the only source for site-specific input. Another important use of the site descriptive model is in the environmental impact assessment. An SDM is an integrated model of geology, thermal properties, rock mechanics, hydrogeology, hydrogeochemistry, bedrock transport properties and a description of the surface system. The site descriptive model compiled in the current report, SDM-Site Laxemar, presents an integrated understanding of the Laxemar-Simpevarp area (with special emphasis on the Laxemar subarea) at the completion of the surface-based investigations, which were conducted during the period 2002 to 2007. A summary is also provided of the abundant underlying data and the discipline specific models that support the site understanding. The description relies heavily on background reports that address, in particular, details of the data analyses and modelling of the different disciplines. The Laxemar-Simpevarp area is located in the province of Smaaland within the municipality of Oskarshamn, about 230 km south of Stockholm. The candidate area for site investigation is located along the shoreline of the strait of Kalmarsund, within a 1.8 billion year old suite of well preserved bedrock belonging to the Transscandinavian Igneous Belt formed during

Site description of Forsmark at completion of the site investigation phase. SDM-Site Forsmark

International Nuclear Information System (INIS)

2008-12-01

The Swedish Nuclear Fuel and Waste Management Co., SKB, has undertaken site characterisation in two different areas, Forsmark and Laxemar-Simpevarp, in order to identify a suitable location for a geological repository of spent nuclear fuel according to the KBS-3 method. The site investigations have been conducted in campaigns, punctuated by data freezes. After each data freeze, the site data have been analysed and modelling has been carried out with the overall purpose to develop a site descriptive model (SDM). The site descriptive model is used by repository engineering to design the underground facility and to develop a repository layout adapted to the site. It is also essential for safety assessment, since the model is the only source for site-specific input. Another important use of the site descriptive model is in the environmental impact assessment. An SDM is an integrated model for geology, thermal properties, rock mechanics, hydrogeology, hydrogeochemistry, bedrock transport properties and a description of the surface system. The site descriptive model compiled in the current report, SDM-Site, presents an integrated understanding of the Forsmark area at the completion of the surface-based investigations, which were conducted at Forsmark during the period 2002 to 2007. It also provides a summary of the abundant underlying data and the discipline-specific models that support the site understanding. The description relies heavily on background reports that address, in particular, details in data analyses and modelling in the different disciplines. The Forsmark area is located in northern Uppland within the municipality of Oesthammar, about 120 km north of Stockholm. The candidate area for site investigation is located along the shoreline of Oeregrundsgrepen, within the north-western part of a major tectonic lens that formed between 1.87 and 1.85 billion years ago during the Svecokarelian orogeny. The candidate area is approximately 6 km long and 2 km wide. The

Eel calcitonin binding site distribution and antinociceptive activity in rats

International Nuclear Information System (INIS)

Guidobono, F.; Netti, C.; Sibilia, V.; Villa, I.; Zamboni, A.; Pecile, A.

1986-01-01

The distribution of binding site for [ 125 I]-eel-calcitonin (ECT) to rat central nervous system, studied by an autoradiographic technique, showed concentrations of binding in the diencephalon, the brain stem and the spinal cord. Large accumulations of grains were seen in the hypothalamus, the amygdala, in the fasciculus medialis prosencephali, in the fasciculus longitudinalis medialis, in the ventrolateral part of the periventricular gray matter, in the lemniscus medialis and in the raphe nuclei. The density of grains in the reticular formation and in the nucleus tractus spinalis nervi trigemini was more moderate. In the spinal cord, grains were scattered throughout the dorsal horns. Binding of the ligand was displaced equally by cold ECT and by salmon CT(sCT), indicating that both peptides bind to the same receptors. Human CT was much weaker than sCT in displacing [ 125 I]-ECT binding. The administration of ECT into the brain ventricles of rats dose-dependently induced a significant and long-lasting enhancement of hot-plate latencies comparable with that obtained with sCT. The antinociceptive activity induced by ECT is compatible with the topographical distribution of binding sites for the peptide and is a further indication that fish CTs are active in the mammalian brain

Mouse papillomavirus infections spread to cutaneous sites with progression to malignancy.

Science.gov (United States)

Cladel, Nancy M; Budgeon, Lynn R; Cooper, Timothy K; Balogh, Karla K; Christensen, Neil D; Myers, Roland; Majerciak, Vladimir; Gotte, Deanna; Zheng, Zhi-Ming; Hu, Jiafen

2017-09-25

We report secondary cutaneous infections in the mouse papillomavirus (MmuPV1)/mouse model. Our previous study demonstrated that cutaneous MmuPV1 infection could spread to mucosal sites. Recently, we observed that mucosal infections could also spread to various cutaneous sites including the back, tail, muzzle and mammary tissues. The secondary site lesions were positive for viral DNA, viral capsid protein and viral particles as determined by in situ hybridization, immunohistochemistry and transmission electron microscopy analyses, respectively. We also demonstrated differential viral production and tumour growth at different secondarily infected skin sites. For example, fewer viral particles were detected in the least susceptible back tissues when compared with those in the infected muzzle and tail, although similar amounts of viral DNA were detected. Follow-up studies demonstrated that significantly lower amounts of viral DNA were packaged in the back lesions. Lavages harvested from the oral cavity and lower genital tracts were equally infectious at both cutaneous and mucosal sites, supporting the broad tissue tropism of this papillomavirus. Importantly, two secondary skin lesions on the forearms of two mice displayed a malignant phenotype at about 9.5 months post-primary infection. Therefore, MmuPV1 induces not only dysplasia at mucosal sites such as the vagina, anus and oral cavity but also skin carcinoma at cutaneous sites. These findings demonstrate that MmuPV1 mucosal infection can be spread to cutaneous sites and suggest that the model could serve a useful role in the study of the viral life cycle and pathogenesis of papillomavirus.

Application of NUSS siting guidelines for nuclear power plants in India

International Nuclear Information System (INIS)

Soman, S.D.; Sarma, T.P.

1985-01-01

The nuclear power profile in India envisages generation of 10,000 MW(e) by the year 2000. In the selection of sites for this purpose, safety considerations based on the NUSS siting code were developed, keeping in mind the conditions existing in the country. The paper reviews the similarity of approach between NUSS documents on siting and guidelines being adopted in India, given different site characteristics such as population distribution, seismicity, flooding and external man-induced events. However, in a few aspects, difficulties were experienced in adopting NUSS requirements in toto. This was primarily because of differences in the national systems, lack of instrumentally recorded data, and an inadequate scientific investigational base in fields such as seismology and hydrology. This paper discusses two such examples and how the difficulties have been overcome in arriving at guidelines which would be in the same spirit as the siting recommendations of the NUSS programme. The first example relates to identification of capable fault and the second concerns the flooding arising out of failure of upstream dams. In addition, the inadequacy and inappropriateness of some of the models given in the NUSS siting guides are also discussed. The paper also considers the current status of accident evaluation of PHWR (235 MW(e)) units being built in the country. In the light of this evaluation, the siting criteria and the emergency measures planned at the sites are reviewed for their compatibility. The emergency measures planned take full cognizance of the ICRP recommendations related to the protection of population in the event of a major reactor accident. (author)

Richness and composition of gall-inducing arthropods at Coiba National Park, Panama

OpenAIRE

Nieves-Aldrey, José Luis; Enrique Medianero, Alicia Ibáñez

2008-01-01

Interest in studying galls and their arthropods inducers has been growing rapidly in the last two decades. However, the Neotropical region is probably the least studied region for gall-inducing arthropods. A study of the richness and composition of gall-inducing arthropods was carried out at Coiba National Park in the Republic of Panama. Field data come from samples obtained between August 1997 and September 1999, with three (two-week long) more intensive samplings. Seventeen sites, represent...

On-site and off-site activities

International Nuclear Information System (INIS)

Martin, H.D.

1986-01-01

Design principles for NPP training programs. Effects of NPP contracts. Effects of domestic industrial activities. The role of international bodies. Requirements for on-site training. Training abroad, technical, financial and social aspects. Training center on-site, an evaluation. (orig.)

Region 9 NPL Sites (Superfund Sites 2013)

Science.gov (United States)

NPL site POINT locations for the US EPA Region 9. NPL (National Priorities List) sites are hazardous waste sites that are eligible for extensive long-term cleanup under the Superfund program. Eligibility is determined by a scoring method called Hazard Ranking System. Sites with high scores are listed on the NPL. The majority of the locations are derived from polygon centroids of digitized site boundaries. The remaining locations were generated from address geocoding and digitizing. Area covered by this data set include Arizona, California, Nevada, Hawaii, Guam, American Samoa, Northern Marianas and Trust Territories. Attributes include NPL status codes, NPL industry type codes and environmental indicators. Related table, NPL_Contaminants contains information about contaminated media types and chemicals. This is a one-to-many relate and can be related to the feature class using the relationship classes under the Feature Data Set ENVIRO_CONTAMINANT.

On-Site or Off-Site Renewable Energy Supply Options?

DEFF Research Database (Denmark)

Marszal, Anna Joanna; Heiselberg, Per; Jensen, Rasmus Lund

2012-01-01

The concept of a Net Zero Energy Building (Net ZEB) encompasses two options of supplying renewable energy, which can offset energy use of a building, in particular on-site or off-site renewable energy supply. Currently, the on-site options are much more popular than the off-site; however, taking...... into consideration the limited area of roof and/or façade, primarily in the dense city areas, the Danish weather conditions, the growing interest and number of wind turbine co-ops, the off-site renewable energy supply options could become a meaningful solution for reaching ‘zero’ energy goal in the Danish context...... five technologies, i.e., two on-site options: (1) photovoltaic, (2) micro combined heat and power, and three off-site options: (1) off-site windmill, (2) share of a windmill farm and (3) purchase of green energy from the 100% renewable utility grid. The results indicate that in case of the on...

Vasomotor response to cold stimulation in human capsaicin-induced hyperalgesic area.

Science.gov (United States)

Pud, Dorit; Andersen, Ole Kaeseler; Arendt-Nielsen, Lars; Eisenberg, Elon; Yarnitsky, David

2005-07-01

Cooling the skin induces sympathetically driven vasoconstriction, with some vasoparalytic dilatation at the lowest temperatures. Neurogenic inflammation, on the other hand, entails vasodilatation. In this study we investigated the balance between vasoconstriction and vasodilatation in an area of experimentally induced secondary hyperalgesia (2 degrees HA), in response to low-temperature stimulations. Fourteen healthy volunteers were exposed to three 30-s long cold stimuli (20, 10, and 0 degrees C) applied, at three adjacent sites, before (baseline) and 8 min after intradermal injection of 50 microg capsaicin to the volar forearm. The cold stimuli were applied distally to the injection site within the 2 degrees HA. Blood flux (BF) and skin temperatures were measured at four different regions (proximally, and distally to the capsaicin injection and at the 0, 10, and 20 degrees C thermode sites) all within the 2 degrees HA. The vascular measurements were conducted five times. Results showed a marked increase in BF after baseline cold stimulation (Peffect (elevated BF) was found following the capsaicin injection compared with baseline for all regions (Pcooled area was dilated by 450+/-5.1%; The vasoconstrictive effect for the 10 and 20 degrees C did not overcome the capsaicin vasodilatation, but did reduce it, with dilatation of 364+/-7.0% and 329+/-7.3%, respectively. For 0 degrees C, a dilatation of 407+/-6.5% was seen. It is concluded that in this experimental model, and potentially in the equivalent clinical syndromes, vasodilatation induced by the inflammation is only slightly reduced by cold stimulation such that it is still dominant, despite some cold-induced vasoconstriction.

Structure of Dioclea virgata lectin: relations between carbohydrate binding site and nitric oxide production

International Nuclear Information System (INIS)

Delatorre, P.; Gadelha, C.A.A.; Santi-Gadelha, T.; Nobrega, R.B.; Rocha, B.A.M.; Nascimento, K.S.; Naganao, C.S.; Sampaio, A.H.; Cavada, B.S.; Pires, A.F.; Assreuy, A.M.S.

2012-01-01

Full text: Lectins are proteins/glycoproteins with at least one noncatalytic domain binding reversibly to specific monosaccharides or oligosaccharides. By binding to carbohydrate moieties on the cell surface, lectins participate in a range of cellular processes without changing the properties of the carbohydrates involved. The lectin of Dioclea virgata (DvirL), both native and complexed with X-man, was submitted to X-ray diffraction analysis and the crystal structure was compared to that of other Diocleinae lectins in order to better understand differences in biological proper- ties, especially with regard to the ability of lectins to induce nitric oxide (NO) production. The DvirL diffraction analysis revealed that both the native crystal and the X-Man-complexed form are orthorhombic and belong to space group I222. The cell parameters were: a=65.4 , b=86.6 and c=90.2 (native structure), and a=61.89 , b=87.67 and c=88.78 (X-Man-complexed structure). An association was observed between the volume of the carbohydrate recognition domain (CRD), the ability to induce NO production and the relative positions of Tyr12, Arg228 and Leu99. Thus, differences in biological activity induced by Diocleinae lectins are related to the configuration of amino acid residues in the carbohydrate binding site and to the structural conformation of subsequent regions capable of influencing site-ligand interactions. In conclusion, the ability of Diocleinae lectins to induce NO production depends on CRD configuration. (author)

Flow-induced vibration -- 1994. PVP-Volume 273

International Nuclear Information System (INIS)

Au-Yang, M.K.; Fujita, K.

1994-01-01

Flow-induced vibration is a subject of practical interest to many engineering disciplines, including the power generation, process, and petrochemical industries. In the nuclear industry, flow-induced vibration reaches a higher level of concern because of safety issues and the huge cost associated with down time and site repair. Not surprisingly, during the last 25 years a tremendous amount of effort has been spent in the study of flow-induced vibration phenomena related to nuclear plant components, notably nuclear steam generator tube banks and nuclear fuel bundles. Yet, in spite of this concentrated effort, the industry is still not free from flow-induced vibration-related problems. This explains why in this volume almost half of the papers address the issue of cross-flow induced vibration in tube bundles, with applications to the nuclear steam generator and nuclear fuel bundles in mind. Unlike 10 or 15 years ago, when flow-induced vibration studies almost always involved experimentation and empirical studies, the advent of high-speed computers has enabled numerical calculation and simulation of this complex phenomenon to take place. Separate abstracts were prepared for 27 papers in this volume

NMDA receptor glycine modulatory site in the ventral tegmental area regulates the acquisition, retrieval, and reconsolidation of cocaine reward memory.

Science.gov (United States)

Zhou, Shuang-jiang; Xue, Li-fen; Wang, Xue-yi; Jiang, Wen-gao; Xue, Yan-xue; Liu, Jian-feng; He, Yin-yin; Luo, Yi-xiao; Lu, Lin

2012-05-01

Accumulating clinical and preclinical studies have shown that the memories of the rewarding effects of drugs and their paired cues may contribute to relapse and persistent cocaine use. Glutaminergic actions in the ventral tegmental area (VTA) have been shown to regulate the rewarding effect of drugs and conditioned responses to drug-associated cues, but the role of the VTA in the acquisition, retrieval, and reconsolidation of cocaine cues is not yet known. In the present study, we used 7-chlorothiokynurenic acid (7-CTKA), an N-methyl-D-aspartate (NMDA) receptor glycine modulatory site antagonist with no rewarding effects, to examine the role of the NMDA receptor glycine modulatory site in the acquisition, retrieval, and reconsolidation of cocaine-related reward memory using the conditioned place preference (CPP) paradigm. Separate groups of Sprague-Dawley rats were trained to acquire cocaine-induced CPP. Vehicle or 7-CTKA was microinjected into the VTA or substantia nigra (SN) (5 μg/μl) at different time points: 10 min before each CPP training session (acquisition), 10 min before the reactivation of CPP (retrieval), and immediately after the reactivation of CPP (reconsolidation). Cocaine-induced CPP was retested 24 h and 1 and 2 weeks after 7-CTKA administration. 7-CTKA microinjected into the VTA, but not SN, significantly impaired the acquisition, retrieval, and reconsolidation of cocaine-induced CPP without affecting cocaine-induced locomotion. Our findings suggest that the NMDA receptor glycine modulatory site in the VTA plays a major role in cocaine reward memory, and NMDA receptor glycine site antagonists may be potential pharmacotherapies for the management of relapse.

Mapping the Binding Site for Escitalopram and Paroxetine in the Human Serotonin Transporter Using Genetically Encoded Photo-Cross-Linkers

DEFF Research Database (Denmark)

Rannversson, Hafsteinn; Andersen, Jacob; Bang-Andersen, Benny

2017-01-01

amber codon suppression in hSERT to encode the photo-cross-linking unnatural amino acid p-azido-l-phenylalanine into the suggested high- and low-affinity binding sites. We then employ UV-induced cross-linking with azF to map the binding site of escitalopram and paroxetine, two prototypical selective...... serotonin reuptake inhibitors (SSRIs). We find that the two antidepressant drugs exclusively cross-link to azF incorporated at the high-affinity binding site of hSERT, while cross-linking is not observed at the low-affinity binding site. Combined with previous homology models and recent structural data on h...

Off-site interaction effect in the Extended Hubbard Model with the SCRPA method

International Nuclear Information System (INIS)

Harir, S; Bennai, M; Boughaleb, Y

2007-01-01

The self consistent random phase approximation (SCRPA) and a direct analytical (DA) method are proposed to solve the Extended Hubbard Model (EHM) in one dimension (1D). We have considered an EHM including on-site and off-site interactions for closed chains in 1D with periodic boundary conditions. The comparison of the SCRPA results with the ones obtained by a DA approach shows that the SCRPA treats the problem of these closed chains in a rigorous manner. The analysis of the nearest-neighbour repulsion effect on the dynamics of our closed chains shows that this repulsive interaction between the electrons of the neighbouring atoms induces supplementary conductivity, since, the SCRPA energygap vanishes when these closed chains are governed by a strong repulsive on-site interaction and intermediate nearest-neighbour repulsion

Earthquake-induced liquefaction in Ferland, Quebec

International Nuclear Information System (INIS)

Tuttle, M.; Seeber, L.

1991-02-01

Detailed geological investigations are under way at a number of liquefaction sites in the Ferland-Boilleau valley, Quebec, where sand boils, ground cracks and liquefaction-related damages to homes were documented immediately following the Ms=6.0, Mblg=6.5 Saguenay earthquake of November 25, 1988. To date, results obtained from these subsurface investigations of sand boils at two sites in Ferland, located about 26 km from the epicentre, indicate that: the Saguenay earthquake induced liquefaction in late-Pleistocene and Holocene sediments which was recorded as sand dikes, sills and vents in near-surface sediments and soils; earthquake-induced liquefaction and ground failure have occurred in this area at least three times in the past 10,000 years; and, the size and morphology of liquefaction features and the liquefaction susceptibility of source layers of the features may be indicative of the intensity of ground shaking. These preliminary results are very promising and suggest that with continued research liquefaction features will become a useful tool in glaciated terrains, such as northeastern North America, for determining not only the timing and location but also the size of past earthquakes

Poisoning Experiments Aimed at Discriminating Active and Less-Active Sites of Silica-Supported Tantalum Hydride for Alkane Metathesis

KAUST Repository

Saggio, Guillaume

2010-10-04

Only 50% of the silica-supported tantalum hydride sites are active in the metathesis of propane. Indeed, more than 45% of the tantalum hydride can be eliminated by a selective oxygen poisoning of inactive sites with no significant decrease in the global turnover. Conversely, cyclopentane induces no such selective poisoning. Hence, the active tantalum hydride sites that show greater resistance to oxygen poisoning correspond to the νTa-H bands of higher wavenumbers, particularly that at 1860cm-1. These active tantalum hydride sites should correspond to tris- or monohydride species relatively far from silica surface oxygen atoms. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A novel functional site of extracellular matrix metalloproteinase inducer (EMMPRIN) that limits the migration of human uterine cervical carcinoma cells.

Science.gov (United States)

Sato, Takashi; Watanabe, Mami; Hashimoto, Kei; Ota, Tomoko; Akimoto, Noriko; Imada, Keisuke; Nomizu, Motoyoshi; Ito, Akira

2012-01-01

EMMPRIN (extracellular matrix metalloproteinase inducer)/CD147, a membrane-bound glycoprotein with two extracellular loop domains (termed loops I and II), progresses tumor invasion and metastasis by increasing the production of matrix metalloproteinase (MMP) in peritumoral stoma cells. EMMPRIN has also been associated with the control of migration activity in some tumor cells, but little is known about how EMMPRIN regulates tumor cell migration. In the present study, EMMPRIN siRNA suppressed the gene expression and production of EMMPRIN in human uterine cervical carcinoma SKG-II cells. An in vitro scratch wound assay showed enhancement of migration of EMMPRIN-knockdown SKG-II cells. In addition, the SKG-II cell migration was augmented by adding an E. coli-expressed human EMMPRIN mutant with two extracellular loop domains (eEMP-I/II), which bound to the cell surface of SKG-II cells. However, eEMP-I/II suppressed the native EMMPRIN-mediated augmentation of proMMP-1/procollagenase-1 production in a co-culture of the SKG-II cells and human uterine cervical fibroblasts, indicating that the augmentation of SKG-II cell migration resulted from the interference of native EMMPRIN functions by eEMP-I/II on the cell surface. Furthermore, a systematic peptide screening method using nine synthetic EMMPRIN peptides coding the loop I and II domains (termed EM1-9) revealed that EM9 (170HIENLNMEADPGQYR184) facilitated SKG-II cell migration. Moreover, SKG-II cell migration was enhanced by administration of an antibody against EM9, but not EM1 which is a crucial site for the MMP inducible activity of EMMPRIN. Therefore, these results provide novel evidence that EMMPRIN on the cell surface limits the cell migration of human uterine cervical carcinoma cells through 170HIENLNMEADPGQYR184 in the loop II domain. Finally, these results should provide an increased understanding of the functions of EMMPRIN in malignant cervical carcinoma cells, and could contribute to the development of

Rivastigmine alleviates experimentally induced colitis in mice and rats by acting at central and peripheral sites to modulate immune responses.

Directory of Open Access Journals (Sweden)

Helena Shifrin

Full Text Available The cholinergic anti-inflammatory system and α7 nicotinic receptors in macrophages have been proposed to play a role in neuroimmunomodulation and in the etiology of ulcerative colitis. We investigated the ability of a cholinesterase (ChE inhibitor rivastigmine, to improve the pathology of ulcerative colitis by increasing the concentration of extracellular acetylcholine in the brain and periphery. In combination with carbachol (10 µM, rivastigmine (1 µM significantly decreased the release of nitric oxide, TNF-α, IL-1β and IL-6 from lipopolysaccharide-activated RAW 264.7 macrophages and this effect was abolished by α7 nicotinic receptor blockade by bungarotoxin. Rivastigmine (1 mg/kg but not (0.5 mg/kg, injected subcutaneously once daily in BALB/c mice with colitis induced by 4% dextran sodium sulphate (DSS, reduced the disease activity index (DAI by 60% and damage to colon structure. Rivastigmine (1 mg/kg also reduced myeloperoxidase activity and IL-6 by >60%, and the infiltration of CD11b expressing cells by 80%. These effects were accompanied by significantly greater ChE inhibition in cortex, brain stem, plasma and colon than that after 0.5 mg/kg. Co-administration of rivastigmine (1 mg/kg with the muscarinic antagonist scopolamine significantly increased the number of CD11b expressing cells in the colon but did not change DAI compared to those treated with rivastigmine alone. Rivastigmine 1 and 2 mg given rectally to rats with colitis induced by rectal administration of 30 mg dintrobezene sulfonic acid (DNBS also caused a dose related reduction in ChE activity in blood and colon, the number of ulcers and area of ulceration, levels of TNF-α and in MPO activity. The study revealed that the ChE inhibitor rivastigmine is able to reduce gastro-intestinal inflammation by actions at various sites at which it preserves ACh. These include ACh released from vagal nerve endings that activates alpha7 nicotinic receptors on circulating macrophages

Nuclear power plant site evaluation using site population-meterology factor

International Nuclear Information System (INIS)

Rho, B.H.; Kang, C.H.

1982-01-01

In this paper, as a site evaluation technique, SPNF(Site Population Neteorology Factor) which is modified from SPF(Site Population Factor) of the USNRC model, is defined from site population and meteorology data in order to consider the radiological impacts to the population at large from the atmospheric dispersion of the radioactive effluents released during routine plant operation as well as accidental conditions. The SPMF model proved its propriety from the comparison of SPMF and SPF for Kori site. The relative suitability of Korean sites to the U.S. sites have been also examined using SPF. (Author)

Ectopic expression of AID in a non-B cell line triggers A:T and G:C point mutations in non-replicating episomal vectors.

Directory of Open Access Journals (Sweden)

Tihana Jovanic

Full Text Available Somatic hypermutation (SHM of immunoglobulin genes is currently viewed as a two step process initiated by the deamination of deoxycytidine (C to deoxyuridine (U, catalysed by the activation induced deaminase (AID. Phase 1 mutations arise from DNA replication across the uracil residue or the abasic site, generated by the uracil-DNA glycosylase, yielding transitions or transversions at G:C pairs. Phase 2 mutations result from the recognition of the U:G mismatch by the Msh2/Msh6 complex (MutS Homologue, followed by the excision of the mismatched nucleotide and the repair, by the low fidelity DNA polymerase eta, of the gap generated by the exonuclease I. These mutations are mainly focused at A:T pairs. Whereas in activated B cells both G:C and A:T pairs are equally targeted, ectopic expression of AID was shown to trigger only G:C mutations on a stably integrated reporter gene. Here we show that when using non-replicative episomal vectors containing a GFP gene, inactivated by the introduction of stop codons at various positions, a high level of EGFP positive cells was obtained after transient expression in Jurkat cells constitutively expressing AID. We show that mutations at G:C and A:T pairs are produced. EGFP positive cells are obtained in the absence of vector replication demonstrating that the mutations are dependent only on the mismatch repair (MMR pathway. This implies that the generation of phase 1 mutations is not a prerequisite for the expression of phase 2 mutations.

Uracil excision repair in Mycobacterium tuberculosis cell-free extracts.

Science.gov (United States)

Kumar, Pradeep; Bharti, Sanjay Kumar; Varshney, Umesh

2011-05-01

Uracil excision repair is ubiquitous in all domains of life and initiated by uracil DNA glycosylases (UDGs) which excise the promutagenic base, uracil, from DNA to leave behind an abasic site (AP-site). Repair of the resulting AP-sites requires an AP-endonuclease, a DNA polymerase, and a DNA ligase whose combined activities result in either short-patch or long-patch repair. Mycobacterium tuberculosis, the causative agent of tuberculosis, has an increased risk of accumulating uracils because of its G + C-rich genome, and its niche inside host macrophages where it is exposed to reactive nitrogen and oxygen species, two major causes of cytosine deamination (to uracil) in DNA. In vitro assays to study DNA repair in this important human pathogen are limited. To study uracil excision repair in mycobacteria, we have established assay conditions using cell-free extracts of M. tuberculosis and M. smegmatis (a fast-growing mycobacterium) and oligomer or plasmid DNA substrates. We show that in mycobacteria, uracil excision repair is completed primarily via long-patch repair. In addition, we show that M. tuberculosis UdgB, a newly characterized family 5 UDG, substitutes for the highly conserved family 1 UDG, Ung, thereby suggesting that UdgB might function as backup enzyme for uracil excision repair in mycobacteria. Copyright © 2011 Elsevier Ltd. All rights reserved.

Identification of ATM Protein Kinase Phosphorylation Sites by Mass Spectrometry.

Science.gov (United States)

Graham, Mark E; Lavin, Martin F; Kozlov, Sergei V

2017-01-01

ATM (ataxia-telangiectasia mutated) protein kinase is a key regulator of cellular responses to DNA damage and oxidative stress. DNA damage triggers complex cascade of signaling events leading to numerous posttranslational modification on multitude of proteins. Understanding the regulation of ATM kinase is therefore critical not only for understanding the human genetic disorder ataxia-telangiectasia and potential treatment strategies, but essential for deciphering physiological responses of cells to stress. These responses play an important role in carcinogenesis, neurodegeneration, and aging. We focus here on the identification of DNA damage inducible ATM phosphorylation sites to understand the importance of autophosphorylation in the mechanism of ATM kinase activation. We demonstrate the utility of using immunoprecipitated ATM in quantitative LC-MS/MS workflow with stable isotope dimethyl labeling of ATM peptides for identification of phosphorylation sites.

Reduction of GABA/sub B/ receptor binding induced by climbing fiber degeneration in the rat cerebellum

International Nuclear Information System (INIS)

Kato, K.; Fukuda, H.

1985-01-01

When the rat cerebellar climbing fibers degenerated, as induced by lesioning the inferior olive with 3-acetylpyridine (3-AP), GABA/sub B/ receptor binding determined with 3 H-(+/-)baclofen was reduced in the cerebellum but not in the cerebral cortex of rats. Computer analysis of saturation data revealed two components of the binding sites, and indicated that decrease of the binding in the cerebellum was due to reduction in receptor density, mainly of the high-affinity sites, the B/sub max/ of which was reduced to one-third that in the control animals. In vitro treatment with 3-AP, of the membranes prepared from either the cerebellum or the cerebral cortex, induced no alteration in the binding sites, thereby indicating that the alteration of GABA/sub B/ sites induced by in vivo treatment with 3-AP is not due to a direct action of 3-AP on the receptor. GABA/sub A/ and benzodiazepine receptor binding labelled with 3 H-muscimol and 3 H-diazepam, respectively, in both of brain regions was not affected by destruction of the inferior olive. These results provide evidence that some of the GABA/sub B/ sites but neither GABA/sub A/ nor benzodiazepine receptors in the cerebellum are located at the climbing fiber terminals. 28 references, 4 figures, 2 tables

Motility and chemotaxis mediate the preferential colonization of gastric injury sites by Helicobacter pylori.

Directory of Open Access Journals (Sweden)

Eitaro Aihara

2014-07-01

Full Text Available Helicobacter pylori (H. pylori is a pathogen contributing to peptic inflammation, ulceration, and cancer. A crucial step in the pathogenic sequence is when the bacterium first interacts with gastric tissue, an event that is poorly understood in vivo. We have shown that the luminal space adjacent to gastric epithelial damage is a microenvironment, and we hypothesized that this microenvironment might enhance H. pylori colonization. Inoculation with 106 H. pylori (wild-type Sydney Strain 1, SS1 significantly delayed healing of acetic-acid induced ulcers at Day 1, 7 and 30 post-inoculation, and wild-type SS1 preferentially colonized the ulcerated area compared to uninjured gastric tissue in the same animal at all time points. Gastric resident Lactobacillus spp. did not preferentially colonize ulcerated tissue. To determine whether bacterial motility and chemotaxis are important to ulcer healing and colonization, we analyzed isogenic H. pylori mutants defective in motility (ΔmotB or chemotaxis (ΔcheY. ΔmotB (10(6 failed to colonize ulcerated or healthy stomach tissue. ΔcheY (10(6 colonized both tissues, but without preferential colonization of ulcerated tissue. However, ΔcheY did modestly delay ulcer healing, suggesting that chemotaxis is not required for this process. We used two-photon microscopy to induce microscopic epithelial lesions in vivo, and evaluated accumulation of fluorescently labeled H. pylori at gastric damage sites in the time frame of minutes instead of days. By 5 min after inducing damage, H. pylori SS1 preferentially accumulated at the site of damage and inhibited gastric epithelial restitution. H. pylori ΔcheY modestly accumulated at the gastric surface and inhibited restitution, but did not preferentially accumulate at the injury site. H. pylori ΔmotB neither accumulated at the surface nor inhibited restitution. We conclude that bacterial chemosensing and motility rapidly promote H. pylori colonization of injury sites

Motility and Chemotaxis Mediate the Preferential Colonization of Gastric Injury Sites by Helicobacter pylori

Science.gov (United States)

Aihara, Eitaro; Closson, Chet; Matthis, Andrea L.; Schumacher, Michael A.; Engevik, Amy C.; Zavros, Yana; Ottemann, Karen M.; Montrose, Marshall H.

2014-01-01

Helicobacter pylori (H. pylori) is a pathogen contributing to peptic inflammation, ulceration, and cancer. A crucial step in the pathogenic sequence is when the bacterium first interacts with gastric tissue, an event that is poorly understood in vivo. We have shown that the luminal space adjacent to gastric epithelial damage is a microenvironment, and we hypothesized that this microenvironment might enhance H. pylori colonization. Inoculation with 106 H. pylori (wild-type Sydney Strain 1, SS1) significantly delayed healing of acetic-acid induced ulcers at Day 1, 7 and 30 post-inoculation, and wild-type SS1 preferentially colonized the ulcerated area compared to uninjured gastric tissue in the same animal at all time points. Gastric resident Lactobacillus spp. did not preferentially colonize ulcerated tissue. To determine whether bacterial motility and chemotaxis are important to ulcer healing and colonization, we analyzed isogenic H. pylori mutants defective in motility (ΔmotB) or chemotaxis (ΔcheY). ΔmotB (106) failed to colonize ulcerated or healthy stomach tissue. ΔcheY (106) colonized both tissues, but without preferential colonization of ulcerated tissue. However, ΔcheY did modestly delay ulcer healing, suggesting that chemotaxis is not required for this process. We used two-photon microscopy to induce microscopic epithelial lesions in vivo, and evaluated accumulation of fluorescently labeled H. pylori at gastric damage sites in the time frame of minutes instead of days. By 5 min after inducing damage, H. pylori SS1 preferentially accumulated at the site of damage and inhibited gastric epithelial restitution. H. pylori ΔcheY modestly accumulated at the gastric surface and inhibited restitution, but did not preferentially accumulate at the injury site. H. pylori ΔmotB neither accumulated at the surface nor inhibited restitution. We conclude that bacterial chemosensing and motility rapidly promote H. pylori colonization of injury sites, and thereby biases

Noise-induced hearing loss in small-scale metal industry in Nepal.

Science.gov (United States)

Whittaker, J D; Robinson, T; Acharya, A; Singh, D; Smith, M

2014-10-01

There has been no previous research to demonstrate the risk of noise-induced hearing loss in industry in Nepal. Limited research on occupational noise-induced hearing loss has been conducted within small-scale industry worldwide, despite it being a substantial and growing cause of deafness in the developing world. The study involved a cross-sectional audiometric assessment, with questionnaire-based examinations of noise and occupational history, and workplace noise level assessment. A total of 115 metal workers and 123 hotel workers (control subjects) were recruited. Noise-induced hearing loss prevalence was 30.4 per cent in metal workers and 4.1 per cent in hotel workers, with a significant odds ratio of 10.3. Except for age and time in occupation, none of the demographic factors were significant in predicting outcomes in regression analyses. When adjusted for this finding, and previous noise-exposed occupations, the odds ratio was 13.8. Workplace noise was significantly different between the groups, ranging from 65.3 to 84.7 dBA in metal worker sites, and from 51.4 to 68.6 dBA in the control sites. Metal workers appear to have a greater risk of noise-induced hearing loss than controls. Additional research on occupational noise-induced hearing loss in Nepal and small-scale industry globally is needed.

Integrin Activation Dynamics between the RGD-binding Site and the Headpiece Hinge*

Science.gov (United States)

Puklin-Faucher, Eileen; Vogel, Viola

2009-01-01

Integrins form mechanical links between the extracellular matrix and the cytoskeleton. Although integrin activation is known to be regulated by an allosteric conformational change, which can be induced from the extracellular or intracellular end of the molecule, little is known regarding the sequence of structural events by which signals propagate between distant sites. Here, we reveal with molecular dynamics simulations of the FnIII10-bound αVβ3 integrin headpiece how the binding pocket and interdomain βA/hybrid domain hinge on the distal end of the βA domain are allosterically linked via a hydrophobic T-junction between the middle of the α1 helix and top of the α7 helix. The key results of this study are: 1) that this T-junction is induced by ligand binding and hinge opening, and thus displays bidirectionality; 2) that formation of this junction can be accelerated by ligand-mediated force; and 3) how formation of this junction is inhibited by Ca2+ in place of Mg2+ at the site adjacent to the metal ion-dependent adhesion site (“ADMIDAS”). Together with recent experimental evidence that integrin complexes can form catch bonds (i.e. become strengthened under force), as well as earlier evidence that Ca2+ at the ADMIDAS results in lower binding affinity, these simulations provide a common structural model for the dynamic process by which integrins become activated. PMID:19762919

Superfund and Toxic Release Inventory Sites - MDC_ContaminatedSite

Data.gov (United States)

NSGIC Local Govt | GIS Inventory — A point feature class of open DERM Contaminated sites - see phase code for status of site. Contaminated sites identifies properties where environmental contamination...

Systematic Analysis of Splice-Site-Creating Mutations in Cancer.

Science.gov (United States)

Jayasinghe, Reyka G; Cao, Song; Gao, Qingsong; Wendl, Michael C; Vo, Nam Sy; Reynolds, Sheila M; Zhao, Yanyan; Climente-González, Héctor; Chai, Shengjie; Wang, Fang; Varghese, Rajees; Huang, Mo; Liang, Wen-Wei; Wyczalkowski, Matthew A; Sengupta, Sohini; Li, Zhi; Payne, Samuel H; Fenyö, David; Miner, Jeffrey H; Walter, Matthew J; Vincent, Benjamin; Eyras, Eduardo; Chen, Ken; Shmulevich, Ilya; Chen, Feng; Ding, Li

2018-04-03

For the past decade, cancer genomic studies have focused on mutations leading to splice-site disruption, overlooking those having splice-creating potential. Here, we applied a bioinformatic tool, MiSplice, for the large-scale discovery of splice-site-creating mutations (SCMs) across 8,656 TCGA tumors. We report 1,964 originally mis-annotated mutations having clear evidence of creating alternative splice junctions. TP53 and GATA3 have 26 and 18 SCMs, respectively, and ATRX has 5 from lower-grade gliomas. Mutations in 11 genes, including PARP1, BRCA1, and BAP1, were experimentally validated for splice-site-creating function. Notably, we found that neoantigens induced by SCMs are likely several folds more immunogenic compared to missense mutations, exemplified by the recurrent GATA3 SCM. Further, high expression of PD-1 and PD-L1 was observed in tumors with SCMs, suggesting candidates for immune blockade therapy. Our work highlights the importance of integrating DNA and RNA data for understanding the functional and the clinical implications of mutations in human diseases. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Repeated exposure to Lutzomyia intermedia sand fly saliva induces local expression of interferon-inducible genes both at the site of injection in mice and in human blood.

Science.gov (United States)

Weinkopff, Tiffany; de Oliveira, Camila I; de Carvalho, Augusto M; Hauyon-La Torre, Yazmin; Muniz, Aline C; Miranda, Jose Carlos; Barral, Aldina; Tacchini-Cottier, Fabienne

2014-01-01

During a blood meal, Lutzomyia intermedia sand flies transmit Leishmania braziliensis, a parasite causing tegumentary leishmaniasis. In experimental leishmaniasis, pre-exposure to saliva of most blood-feeding sand flies results in parasite establishment in absence of any skin damages in mice challenged with dermotropic Leishmania species together with saliva. In contrast, pre-immunization with Lu. intermedia salivary gland sonicate (SGS) results in enhanced skin inflammatory exacerbation upon co-inoculation of Lu. intermedia SGS and L. braziliensis. These data highlight potential unique features of both L. braziliensis and Lu. intermedia. In this study, we investigated the genes modulated by Lu. intermedia SGS immunization to understand their potential impact on the subsequent cutaneous immune response following inoculation of both SGS and L. braziliensis. The cellular recruitment and global gene expression profile was analyzed in mice repeatedly inoculated or not with Lu. intermedia. Microarray gene analysis revealed the upregulation of a distinct set of IFN-inducible genes, an immune signature not seen to the same extent in control animals. Of note this INF-inducible gene set was not induced in SGS pre-immunized mice subsequently co-inoculated with SGS and L. braziliensis. These data suggest the parasite prevented the upregulation of this Lu. intermedia saliva-related immune signature. The presence of these IFN-inducible genes was further analyzed in peripheral blood mononuclear cells (PBMCs) sampled from uninfected human individuals living in a L. braziliensis-endemic region of Brazil thus regularly exposed to Lu. intermedia bites. PBMCs were cultured in presence or absence of Lu. intermedia SGS. Using qRT-PCR we established that the IFN-inducible genes induced in the skin of SGS pre-immunized mice, were also upregulated by SGS in PBMCs from human individuals regularly exposed to Lu. intermedia bites, but not in PBMCs of control subjects. These data demonstrate

GHGT-10 : Assessing the integrity of fault- and top seals at CO2 storage sites

NARCIS (Netherlands)

Orlic, B.; Heege J.H. ter; Wassing, B.

2011-01-01

Induced stress changes due to CO2 injection into geological reservoirs can mechanically damage bounding fault- and top seals creating preferential pathways for CO2 migration from the containment or trigger existing faults causing seismic activity at storage sites. In this paper we present

Umatilla Satellite and Release Sites Project : Final Siting Report.

Energy Technology Data Exchange (ETDEWEB)

Montgomery, James M.

1992-04-01

This report presents the results of site analysis for the Umatilla Satellite and Release Sites Project. The purpose of this project is to provide engineering services for the siting and conceptual design of satellite and release facilities for the Umatilla Basin hatchery program. The Umatilla Basin hatchery program consists of artificial production facilities for salmon and steelhead to enhance production in the Umatilla River as defined in the Umatilla master plan approved in 1989 by the Northwest Power Planning Council. Facilities identified in the master plan include adult salmon broodstock holding and spawning facilities, facilities for recovery, acclimation, and/or extended rearing of salmon juveniles, and development of river sites for release of hatchery salmon and steelhead. The historic and current distribution of fall chinook, summer chinook, and coho salmon and steelhead trout was summarized for the Umatilla River basin. Current and future production and release objectives were reviewed. Twenty seven sites were evaluated for the potential and development of facilities. Engineering and environmental attributes of the sites were evaluated and compared to facility requirements for water and space. Site screening was conducted to identify the sites with the most potential for facility development. Alternative sites were selected for conceptual design of each facility type. A proposed program for adult holding facilities, final rearing/acclimation, and direct release facilities was developed.

Cold-induced vasoconstriction at forearm and hand skin sites: the effect of age

OpenAIRE

Kingma, B.R.M.; Frijns, A.J.H.; Saris, W.H.M.; Steenhoven, van, A.A.; Marken Lichtenbelt, van, W.D.

2010-01-01

During mild cold exposure, elderly are at risk of hypothermia. In humans, glabrous skin at the hands is well adapted as a heat exchanger. Evidence exists that elderly show equal vasoconstriction due to local cooling at the ventral forearm, yet no age effects on vasoconstriction at hand skin have been studied. Here, we tested the hypotheses that at hand sites (a) elderly show equal vasoconstriction due to local cooling and (b) elderly show reduced response to noradrenergic stimuli. Skin perfus...

Site decontamination

International Nuclear Information System (INIS)

Bicker, A.E.

1981-01-01

Among the several DOE sites that have been radiologically decontaminated under the auspices of the Nevada Operations Office are three whose physical characteristics are unique. These are the Tatum Dome Test Site (TDTS) near Hattiesburg, Mississippi; a location of mountainous terrain (Pahute Mesa) on the Nevada Test Site; and the GNOME site near Carlsbad, New Mexico. In each case the contamination, the terrain, and the climate conditions were different. This presentation includes a brief description of each site, the methods used to perform radiological surveys, the logistics required to support the decontamination (including health physics and sample analysis), and the specific techniques used to reduce or remove the contamination

Annotation Method (AM): SE36_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE36_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE14_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE14_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE33_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE33_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE12_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE12_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE20_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE20_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE28_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE28_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE11_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE11_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE17_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE17_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE10_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE10_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE25_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE25_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE30_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE30_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE16_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE16_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE29_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE29_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE35_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE35_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE13_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE13_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE26_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE26_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE27_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE27_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE34_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE34_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE15_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE15_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE31_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE31_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Annotation Method (AM): SE32_AM1 [Metabolonote[Archive

Lifescience Database Archive (English)

Full Text Available abase search. Peaks with no hit to these databases are then selected to secondary s...earch using exactMassDB and Pep1000 databases. After the database search processes, each database hits are m...SE32_AM1 PowerGet annotation A1 In annotation process, KEGG, KNApSAcK and LipidMAPS are used for primary dat

Effect of membrane hyperpolarization induced by a K+ channel opener on histamine-induced Ca2+ mobilization in rabbit arterial smooth muscle.

Science.gov (United States)

Watanabe, Y; Suzuki, A; Suzuki, H; Itoh, T

1996-03-01

1. The role of membrane hyperpolarization on agonist-induced contraction was investigated in intact and alpha-toxin-skinned smooth muscles of rabbit mesenteric artery by use of the ATP-sensitive K+ channel opener, (-)-(3S,4R)-4-(N-acetyl-N-hydroxyamino)-6-cyano-3,4-dihydro-2,2- dimethyl-2H-1-benzopyran-3-ol (Y-26763), and either histamine (Hist) or noradrenaline (NA). 2. Hist (3 microM) and NA (10 microM) both produced a phasic, followed by a tonic increase in intracellular Ca2+ concentration ([Ca2+]i) and force. Y-26763 (10 microM) potently inhibited the NA-induced phasic and tonic increase in [Ca2+]i and force. In contrast, Y-26763 attenuated the Hist-induced phasic increase in [Ca2+]i and force but had almost no effect on the tonic response. However, ryanodine-treatment of muscles in order to inhibit the function of intracellular Ca2+ storage sites altered the action of Y-26763 which now attenuated the Hist-induced tonic increase in [Ca2+]i and force in a concentration-dependent manner (at concentrations > 1 microM). Glibenclamide (10 microM) attenuated the inhibitory action of Y-26763. 3. Hist (3 microM) depolarized the smooth muscle cells to the same extent as NA (10 microM). In the absence of either agonist, Y-26763 (over 30 nM) hyperpolarized the membrane and glibenclamide inhibited this hyperpolarization. Y-26763 (10 microM) almost abolished the NA-induced membrane depolarization, but only slightly attenuated the Hist-induced membrane depolarization in which the delta (delta) value (the difference before and after application of Hist) was not modified by any concentration of Y-26763. In ryanodine-treated smooth muscle cells, Y-26763 hyperpolarized the membrane and potently inhibited the membrane depolarization induced by Hist. 4. In ryanodine-treated muscle, Y-26763 had no measurable effect on the Hist-induced [Ca2+]i-force relationship. Y-26763 also had no apparent effect on the myofilament Ca(2+)-sensitivity in the presence of Hist in alpha

Incident laser modulation of a repaired damage site with a rim in fused silica rear subsurface

Institute of Scientific and Technical Information of China (English)

Li Li; Xiang Xia; Zu Xiao-Tao; Yuan Xiao-Dong; He Shao-Bo; Jiang Xiao-Dong; Zheng Wan-Guo

2012-01-01

Local CO2 laser treatment has proved to be an effective method to prevent the 351-nm laser-induced damage sitesin a fused silica surface from exponentially growing,which is responsible for limiting the lifetime of optics in high fluence laser systems.However,the CO2 laser induced ablation crater is often surrounded by a raised rim at the edge,which can also result in the intensification of transmitted ultraviolet light that may damage the downstream optics.In this work,the three-dimensional finite-difference time-domain method is developed to simulate the distribution of electrical field intensity in the vicinity of the CO2 laser mitigated damage site located in the exit subsurface of fused silica.The simulated results show that the repaired damage sites with raised rims cause more notable modulation to the incident laser than those without rims.Specifically,we present a theoretical model of using dimpled patterning to control the rim structure around the edge of repaired damage sites to avoid damage to downstream optics.The calculated results accord well with previous experimental results and the underlying physical mechanism is analysed in detail.

Downregulation of a tumor suppressor RECK by hypoxia through recruitment of HDAC1 and HIF-1alpha to reverse HRE site in the promoter.

Science.gov (United States)

Lee, Kyung Ju; Lee, Kwang Youl; Lee, You Mie

2010-05-01

Reversion-inducing cysteine-rich protein with Kazal motifs (RECK) is a tumor suppressor and the suppression of RECK is induced by Ras or Her-2/neu oncogenes. However, regulation of RECK under hypoxic microenvironment is largely unknown. Here, we identified that hypoxia significantly downregulates RECK mRNA and protein expression using semiquantitative RT-PCR, real-time RT-PCR and western blot analysis. This repression was reversed by the HDAC inhibitor, trichostatin A (TSA) and HIF-1 inhibitor, YC-1. Hypoxia-induced downregulation of RECK was abolished by knockdown of HDAC1 and HIF-1alpha with respective small interfering RNAs (siRNAs), whereas overexpression of HDAC1 and HIF-1alpha suppressed RECK expression similar to the level under hypoxic conditions. Transfection of a deletion mutant of the second reverse HRE (rHRE2, -2345 to -2333) site of RECK promoter completely removed RECK suppression under hypoxia, indicating that the rHRE2 site is responsible for the inhibition of RECK. Chromatin immunoprecipitation and DNA affinity precipitation assays demonstrated that HDAC1 and HIF-1alpha were recruited to the rHRE2 region of RECK promoter under hypoxic conditions, but the treatment of TSA or YC-1 inhibited their binding to the rHRE2 site. Moreover, TSA and YC-1 inhibited hypoxia-induced cancer cell migration, invasion and MMPs secretion. Taken together, we can conclude that hypoxia induces RECK downregulation through the recruitment of HDAC1 and HIF-1alpha to the rHRE2 site in the promoter and the inhibition of hypoxic RECK silencing would be a therapeutic and preventive target for early tumorigenesis. Copyright 2010 Elsevier B.V. All rights reserved.

Histamine ameliorates spatial memory deficits induced by MK-801 infusion into ventral hippocampus as evaluated by radial maze task in rats

Institute of Scientific and Technical Information of China (English)

Li-sha XU; Li-xia YANG; Wei-wei HU; Xiao YU; Li MA; Lu-ying LIU; Er-qing WEI; Zhong CHEN

2005-01-01

Aim: To investigate the role of histamine in memory deficits induced by MK-801 infusion into the ventral hippocampus in rats. Methods: An 8-arm radial maze (4arms baited) was used to assess spatial memory. Results: Bilateral ventral intrahippocampal (ih) infusion of MK-801 (0.3 μg/site), an N-methyl-D-aspartate (NMDA) antagonist, impaired the retrieval process in both working memory and reference memory. Intrahippocampal injection of histamine (25 or 50 ng/site) or intraperitoneal (ip) injection of histidine (25, 50 or 100 mg/kg) markedly ameliorated the spatial memory deficits induced by MK-801. Both the histamine H1 antagonist pyrilamine (0.5 or 1.0 μg/site, ih) and the H2 antagonist cimetidine (2.5 μg/site,ih) abolished the ameliorating effect of histidine (100 mg/kg, ip) on reference memory deficits, but not that on working memory deficits induced by MK-801. Conclusion:The results indicate that histamine in the ventral hippocampus can ameliorate MK-801-induced spatial memory deficits, and that histamine's effect on reference memory is mediated by postsynaptic histamine H1 and H2 receptors.

Bedrock hydrogeology Forsmark. Site descriptive modelling, SDM-Site Forsmark

Energy Technology Data Exchange (ETDEWEB)

Follin, Sven (SF GeoLogic AB, Taeby (Sweden))

2008-12-15

The Swedish Nuclear Fuel and Waste Management Company (SKB) has conducted site investigations at two different locations, the Forsmark and Laxemar-Simpevarp areas, with the objective of siting a final repository for spent nuclear fuel according to the KBS-3 concept. Site characterisation should provide all data required for an integrated evaluation of the suitability of the investigated site and an important component in the characterisation work is the development of a hydrogeological model. The hydrogeological model is used by repository engineering to design the underground facility and to develop a repository layout adapted to the site. It also provides input to the safety assessment. Another important use of the hydrogeological model is in the environmental impact assessment. This report presents the understanding of the hydrogeological conditions of the bedrock at Forsmark reached following the completion of the surface-based investigations and provides a summary of the bedrock hydrogeological model and the underlying data supporting its development. It constitutes the main reference on bedrock hydrogeology for the site descriptive model concluding the surface-based investigations at Forsmark, SDM-site, and is intended to describe the hydraulic properties and hydrogeological conditions of the bedrock at the site and to give the information essential for demonstrating understanding

DNA lability induced by nimustine and ramustine in rat glioma cells.

Science.gov (United States)

Mineura, K; Fushimi, S; Itoh, Y; Kowada, M

1988-01-01

The DNA labile sites induced by two nitrosoureas, nimustine (ACNU) and ramustine (MCNU) synthesised in Japan, have been examined in highly reiterated DNA sequences of rat glioma cells. Reiterated fragments of 167 and 203 base pairs (bp), obtained after Hind III and Hae III restriction endonuclease digestion of rat glioma cells DNA, were used as target DNA sequences to determine the labile sites. In vitro reaction with ACNU and MCNU resulted in scission products corresponding to the locations of guanine. Subsequent piperidine hydrolysis produced more frequent breaks of the phosphodiester bonds at guanine positions, thus forming alkali-labile sites. Images PMID:3236017

Social Media Sites

Science.gov (United States)

Media Sites Site Registration Contact Us Search AF.mil: Home > AF Sites > Social Media Sites Social Media Welcome to the Air Force social media directory! The directory is a one-stop shop of official Air Force social media pages across various social media sites. Social media is all about

Spatial and seasonal heterogeneity of atmospheric particles induced reactive oxygen species in urban areas and the role of water-soluble metals.

Science.gov (United States)

Gali, Nirmal Kumar; Yang, Fenhuan; Jiang, Sabrina Yanan; Chan, Ka Lok; Sun, Li; Ho, Kin-fai; Ning, Zhi

2015-03-01

Adverse health effects are associated with exposure to atmospheric particulate matter (PM), which carry various chemical constituents and induce both exogenous and endogenous oxidative stress. This study investigated the spatial and seasonal variability of PM-induced ROS at four sites with different characteristics in Hong Kong. Cytotoxicity, exogenous and endogenous ROS was determined on a dose and time dependent analysis. Large spatial variation of ROS was observed with fine PM at urban site showing highest ROS levels while coarse PM at traffic site ranks the top. No consistent seasonal difference was observed for ROS levels among all sites. The highly heterogeneous distribution of PM-induced ROS demonstrates the differential capability of PM to produce oxidative stress, and the need to use appropriate metrics as surrogates of exposure instead of PM mass in epidemiologic studies. Several transition metals were found associated with ROS by different degree illustrating the complexity of mechanisms involved. Copyright © 2015 Elsevier Ltd. All rights reserved.

Induced seismicity and the potential for liability under U.S. law

Science.gov (United States)

Cypser, Darlene A.; Davis, Scott D.

1998-04-01

Research by seismologists over the past 30+ years has firmly established that some human activities induce seismicity. Sometimes induced seismicity causes injuries to people or property. The activities which induce seismicity generally involve extraction of energy, or natural resources, or the disposal of wastes. As the human population increases these extraction and disposal activities will increase in number of sites and intensity of effort as the demands become greater and the resources scarcer. With these increases the number and severity of damaging induced earthquakes is likely to increase. Induced seismicity may cause injuries by vibrations or by seismically induced ground failure. In either case compensation for injuries caused by induced seismicity should be paid for by the inducer. In the United States the inducer of damaging seismicity can be made to pay for the harm caused. Liability for damage caused by vibrations can be based on several legal theories: trespass, strict liability, negligence and nuisance. Our research revealed no cases in which an appellate court has upheld or rejected the application of tort liability to an induced earthquake situation. However, there are numerous analogous cases that support the application of these legal theories to induced seismicity. Vibrations or concussions due to blasting or heavy machinery are sometimes viewed as a `trespass' analogous to a physical invasion. In some states activities which induce earthquakes might be considered `abnormally dangerous' activities that require companies engaged in them to pay for injuries the quakes cause regardless of how careful the inducers were. In some circumstances, a court may find that an inducer was negligent in its site selection or in maintenance of the project. If induced seismicity interferes with the use or enjoyment of another's land, then the inducing activity may be a legal nuisance, even if the seismicity causes little physical damage. In most states of the

Chronic idiopathic thrombocytopenic purpura (ITP): site of platelet sequestration and results of splenectomy

International Nuclear Information System (INIS)

Gugliotta, L.; Guarini, A.; Motta, M.R.; Bachetti, G.; Tura, S.; Isacchi, G.; Ciccone, F.; Lattarini, C.; Mazzucconi, M.G.; Mandelli, F.; Baccarani, M.

1981-01-01

51 Cr-platelet kinetics study was performed in 197 patients with chronic ITP after corticosteroid therapy had failed to induce a long lasting remission. The incidence of splenic, spleno-hepatic, hepatic and diffuse platelet sequestration site was 58%, 17%, 6% and 19%, respectively. Splenic and spleno-hepatic sequestration sites were more frequent in patients less than 30 years old and in patients with a platelet count lower than 50 x 10 9 /l. 111 patients were splenectomized shortly after the study. Normalization of the platelet count was obtained more frequently in patients with splenic and spleno-hepatic sequestration than in the others. Labelled platelet sequestration site was the best predictor of the outcome of splenectomy. Platelet kinetics is a non-invasive investigation that should be performed early after the diagnosis of chronic ITP in all patients eligible for splenectomy. (author)

BDP-30, a systemic resistance inducer from Boerhaavia diffusa L ...

Indian Academy of Sciences (India)

2015-01-11

Jan 11, 2015 ... exogenous application of chemicals such as salicylic acid or its synthetic ...... be able to induce a mobile signal that takes effect both at site of application, and ... thesis of its gene and expression in Escherichia coli. J. Biol.

Empirical Ground Motion Characterization of Induced Seismicity in Alberta and Oklahoma

Science.gov (United States)

Novakovic, M.; Atkinson, G. M.; Assatourians, K.

2017-12-01

We develop empirical ground-motion prediction equations (GMPEs) for ground motions from induced earthquakes in Alberta and Oklahoma following the stochastic-model-based method of Atkinson et al. (2015 BSSA). The Oklahoma ground-motion database is compiled from over 13,000 small to moderate seismic events (M 1 to 5.8) recorded at 1600 seismic stations, at distances from 1 to 750 km. The Alberta database is compiled from over 200 small to moderate seismic events (M 1 to 4.2) recorded at 50 regional stations, at distances from 30 to 500 km. A generalized inversion is used to solve for regional source, attenuation and site parameters. The obtained parameters describe the regional attenuation, stress parameter and site amplification. Resolving these parameters allows for the derivation of regionally-calibrated GMPEs that can be used to compare ground motion observations between waste water injection (Oklahoma) and hydraulic fracture induced events (Alberta), and further compare induced observations with ground motions resulting from natural sources (California, NGAWest2). The derived GMPEs have applications for the evaluation of hazards from induced seismicity and can be used to track amplitudes across the regions in real time, which is useful for ground-motion-based alerting systems and traffic light protocols.

Computer simulations of rare earth sites in glass: experimental tests and applications to laser materials

International Nuclear Information System (INIS)

Weber, M.J.

1984-11-01

Computer simulations of the microscopic structure of BeF 2 glasses using molecular dynamics are reviewed and compared with x-ray and neutron diffraction, EXAFS, NMR, and optical measurements. Unique information about the site-to-site variations in the local environments of rare earth ions is obtained using optical selective excitation and laser-induced fluorescence line-narrowing techniques. Applications and limitations of computer simulations to the development of laser glasses and to predictions of other static and dynamic properties of glasses are discussed. 35 references, 2 figures, 2 tables

Generation of induced pluripotent stem cells from the pig

Science.gov (United States)

The value of stem cells has become increasingly evident in recent years with the advent of genetic engineering tools that allow site-specific modifications to the genome. The use of stem cells to induce modifications has several potential benefits for the livestock industry including improving anim...

Site operations

International Nuclear Information System (INIS)

House, W.B.; Ebenhack, D.G.

1989-01-01

This chapter is a discussion of the management and operations practices used at the Barnwell Waste Management Facility in Barnwell, SC. The following topics are discussed: (1) Waste receiving and inspection, including manifest and certificates of compliance, radiological surveys, disposition of nonconforming items, and decontamination and disposition of secondary waste streams; (2) Waste disposal, including Title 10 CFR 61 requirements, disposal area evaluations, shipment offloading, container emplacement, and radiation protection; (3) Trench closure, including trench backfilling, trench capping, and permanent markers; (4) Site maintenance and stabilization, including trench maintenance, surface water management, and site closure activities; (5) Site monitoring programs, including operational monitoring, and environmental monitoring program; (6) Personnel training and qualifications, including basic training program, safety training program, special skills training, and physical qualifications; (7) Records management, including waste records, personnel training records, personnel dosimetry records, site monitoring records, trench qualification and construction records, and site drawings and stabilization records; (8) Site security; (9) Emergency response plans; and (10) Quality assurance

The abdominal skin of female Sprague-Dawley rats is more sensitive than the back skin to drug-induced phototoxicity.

Science.gov (United States)

Kuga, Kazuhiro; Yasuno, Hironobu; Sakai, Yumi; Harada, Yumiko; Shimizu, Fumi; Miyamoto, Yumiko; Takamatsu, Yuki; Miyamoto, Makoto; Sato, Keiichiro

2017-11-01

In vivo phototoxicity studies are important to predict drug-induced phototoxicity in humans; however, a standard methodology has not established. To determine differences in sensitivity to drug-induced phototoxicity among various skin sites, we evaluated phototoxic reactions in the back and abdominal skin of female Sprague-Dawley rats orally dosed with phototoxic drugs (pirfenidone, 8-methoxysoraren, doxycycline, and lomefloxacin) or a non-phototoxic drug (gatifloxacin) followed by solar-simulated light irradiation comprising 18J/cm 2 ultraviolet A. Tissue reactions were evaluated by macroscopic and microscopic examination and immunohistochemistry for γ-H2AX, and tissue concentrations of pirfenidone, doxycycline, and lomefloxacin were measured by tandem mass spectrometry. In addition, the thicknesses of the skin layers at both sites were measured in drug-naïve rats. The abdominal skin showed more severe reactions to all phototoxic drugs than the back skin, whereas the minimal erythema dose in drug-naïve rats and skin concentrations of each drug were comparable between the sites. Furthermore, histopathological lesions and γ-H2AX-positive cells in the abdominal skin were detected in deeper layers than in the back skin. The stratum corneum and dermis in the abdominal skin were significantly thinner than in the back skin, indicating a difference in the depth of light penetration and potentially contributing to the site differences observed in sensitivity to phototoxicity. Gatifloxacin did not induce any phototoxic reactions at either site. In conclusion, the abdominal skin is more sensitive to drug-induced phototoxicity than the back skin and may represent a preferable site for irradiation in this rat phototoxicity model. Copyright © 2017 Elsevier Inc. All rights reserved.

Infrared Thermography in Serotonin-Induced Itch Model in Rats

DEFF Research Database (Denmark)

Jasemian, Yousef; Gazerani, Parisa; Dagnæs-Hansen, Frederik

2012-01-01

The study validated the application of infrared thermography in a serotonin-induced itch model in rats since the only available method in animal models of itch is the count of scratching bouts. Twenty four adult Sprague-Dawley male rats were used in 3 experiments: 1) local vasomotor response...... with no scratching reflex was investigated. Serotonin elicited significant scratching and lowered the local temperature at the site of injection. A negative dose-temperature relationship of serotonin was found by thermography. Vasoregulation at the site of serotonin injection took place in the absence of scratching...

Intrapulmonary haematoma induced by Swan-Ganz-catherisation

International Nuclear Information System (INIS)

Supra, D.; Jend, H.H.; Oesterreich, F.U.

1989-01-01

A case of circumscribed intrapulmonary haematoma induced by a balloon-tipped catheter is presented in a 56 year old patient with coronary heart disease. The bleeding site appeared as a pulmonary circular focus or coin-shaped lesion on the chest radiograph. The authors discuss possible causes and risk factors of intrapulmonary haematomas and differential diagnosis of pulmonary coin-shaped lesions. (orig.) [de

Prediction of allosteric sites on protein surfaces with an elastic-network-model-based thermodynamic method.

Science.gov (United States)

Su, Ji Guo; Qi, Li Sheng; Li, Chun Hua; Zhu, Yan Ying; Du, Hui Jing; Hou, Yan Xue; Hao, Rui; Wang, Ji Hua

2014-08-01

Allostery is a rapid and efficient way in many biological processes to regulate protein functions, where binding of an effector at the allosteric site alters the activity and function at a distant active site. Allosteric regulation of protein biological functions provides a promising strategy for novel drug design. However, how to effectively identify the allosteric sites remains one of the major challenges for allosteric drug design. In the present work, a thermodynamic method based on the elastic network model was proposed to predict the allosteric sites on the protein surface. In our method, the thermodynamic coupling between the allosteric and active sites was considered, and then the allosteric sites were identified as those where the binding of an effector molecule induces a large change in the binding free energy of the protein with its ligand. Using the proposed method, two proteins, i.e., the 70 kD heat shock protein (Hsp70) and GluA2 alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor, were studied and the allosteric sites on the protein surface were successfully identified. The predicted results are consistent with the available experimental data, which indicates that our method is a simple yet effective approach for the identification of allosteric sites on proteins.

Female site-specific transposase-induced recombination: a high-efficiency method for fine mapping mutations on the X chromosome in Drosophila.

OpenAIRE

Marcus, Jeffrey M

2003-01-01

P-element transposons in the Drosophila germline mobilize only in the presence of the appropriate transposase enzyme. Sometimes, instead of mobilizing completely, P elements will undergo site-specific recombination with the homologous chromosome. Site-specific recombination is the basis for male recombination mapping, since the male germline does not normally undergo recombination. Site-specific recombination also takes place in females, but this has been difficult to study because of the obs...

Superconducting correlations in the one-band Hubbard model with intermediate on-site and weak attractive intersite interactions

International Nuclear Information System (INIS)

Jain, K.P.; Ramakumar, R.; Chancey, C.C.

1990-01-01

In this paper, we analyze a simple extended Hubbard model with an intermediate on-site interaction (both repulsive and attractive) and a weak intersite attractive interaction. Following Hubbard decoupling approximations and introducing Hubbard subband operators, we obtain a generalized gap function for singlet s-wave pairing that explicitly depends on the Hubbard subband energies. For the on-site repulsive-interaction case, we find that the superconductivity is not destroyed in the intermediate-interaction regime, contrary to the prediction of a Hartree-Fock mean-field treatment. The essential consequence of the on-site repulsion is the formation of the Hubbard subbands separated by the Mott-Hubbard gap, and it is within these subbands that pairing induced by the intersite interaction occurs. For the attractive on-site interaction case, the on-site pairing amplitude is found to be proportional to the bandwidth, and the gap function has contributions from both on-site and intersite pairing. The relevance of the model to high-temperature superconductivity is discussed

Structure-based stabilization of HIV-1 gp120 enhances humoral immune responses to the induced co-receptor binding site.

Directory of Open Access Journals (Sweden)

Barna Dey

2009-05-01

Full Text Available The human immunodeficiency virus type 1 (HIV-1 exterior envelope glycoprotein, gp120, possesses conserved binding sites for interaction with the primary virus receptor, CD4, and also for the co-receptor, generally CCR5. Although gp120 is a major target for virus-specific neutralizing antibodies, the gp120 variable elements and its malleable nature contribute to evasion of effective host-neutralizing antibodies. To understand the conformational character and immunogenicity of the gp120 receptor binding sites as potential vaccine targets, we introduced structure-based modifications to stabilize gp120 core proteins (deleted of the gp120 major variable regions into the conformation recognized by both receptors. Thermodynamic analysis of the re-engineered core with selected ligands revealed significant stabilization of the receptor-binding regions. Stabilization of the co-receptor-binding region was associated with a marked increase in on-rate of ligand binding to this site as determined by surface plasmon resonance. Rabbit immunization studies showed that the conformational stabilization of core proteins, along with increased ligand affinity, was associated with strikingly enhanced humoral immune responses against the co-receptor-binding site. These results demonstrate that structure-based approaches can be exploited to stabilize a conformational site in a large functional protein to enhance immunogenic responses specific for that region.

Confidence assessment. Site-descriptive modelling SDM-Site Laxemar

International Nuclear Information System (INIS)

2009-06-01

The objective of this report is to assess the confidence that can be placed in the Laxemar site descriptive model, based on the information available at the conclusion of the surface-based investigations (SDM-Site Laxemar). In this exploration, an overriding question is whether remaining uncertainties are significant for repository engineering design or long-term safety assessment and could successfully be further reduced by more surface-based investigations or more usefully by explorations underground made during construction of the repository. Procedures for this assessment have been progressively refined during the course of the site descriptive modelling, and applied to all previous versions of the Forsmark and Laxemar site descriptive models. They include assessment of whether all relevant data have been considered and understood, identification of the main uncertainties and their causes, possible alternative models and their handling, and consistency between disciplines. The assessment then forms the basis for an overall confidence statement. The confidence in the Laxemar site descriptive model, based on the data available at the conclusion of the surface based site investigations, has been assessed by exploring: - Confidence in the site characterization data base, - remaining issues and their handling, - handling of alternatives, - consistency between disciplines and - main reasons for confidence and lack of confidence in the model. Generally, the site investigation database is of high quality, as assured by the quality procedures applied. It is judged that the Laxemar site descriptive model has an overall high level of confidence. Because of the relatively robust geological model that describes the site, the overall confidence in the Laxemar Site Descriptive model is judged to be high, even though details of the spatial variability remain unknown. The overall reason for this confidence is the wide spatial distribution of the data and the consistency between

Confidence assessment. Site-descriptive modelling SDM-Site Laxemar

Energy Technology Data Exchange (ETDEWEB)

2008-12-15

The objective of this report is to assess the confidence that can be placed in the Laxemar site descriptive model, based on the information available at the conclusion of the surface-based investigations (SDM-Site Laxemar). In this exploration, an overriding question is whether remaining uncertainties are significant for repository engineering design or long-term safety assessment and could successfully be further reduced by more surface-based investigations or more usefully by explorations underground made during construction of the repository. Procedures for this assessment have been progressively refined during the course of the site descriptive modelling, and applied to all previous versions of the Forsmark and Laxemar site descriptive models. They include assessment of whether all relevant data have been considered and understood, identification of the main uncertainties and their causes, possible alternative models and their handling, and consistency between disciplines. The assessment then forms the basis for an overall confidence statement. The confidence in the Laxemar site descriptive model, based on the data available at the conclusion of the surface based site investigations, has been assessed by exploring: - Confidence in the site characterization data base, - remaining issues and their handling, - handling of alternatives, - consistency between disciplines and - main reasons for confidence and lack of confidence in the model. Generally, the site investigation database is of high quality, as assured by the quality procedures applied. It is judged that the Laxemar site descriptive model has an overall high level of confidence. Because of the relatively robust geological model that describes the site, the overall confidence in the Laxemar Site Descriptive model is judged to be high, even though details of the spatial variability remain unknown. The overall reason for this confidence is the wide spatial distribution of the data and the consistency between

Probabilistic siting analysis of nuclear power plants emphasizing atmospheric dispersion of radioactive releases and radiation-induced health effects

International Nuclear Information System (INIS)

Savolainen, Ilkka

1980-01-01

A presentation is made of probabilistic evaluation schemes for nuclear power plant siting. Effects on health attributable to ionizing radiation are reviewed, for the purpose of assessment of the numbers of the most important health effect cases in light-water reactor accidents. The atmospheric dispersion of radioactive releases from nuclear power plants is discussed, and there is presented an environmental consequence assessment model in which the radioactive releases and atmospheric dispersion of the releases are treated by the application of probabilistic methods. In the model, the environmental effects arising from exposure to radiation are expressed as cumulative probability distributions and expectation values. The probabilistic environmental consequence assessment model has been applied to nuclear power plant site evaluation, including risk-benefit and cost-benefit analyses, and the comparison of various alternative sites. (author)

Modeling Shear Induced Von Willebrand Factor Binding to Collagen

Science.gov (United States)

Dong, Chuqiao; Wei, Wei; Morabito, Michael; Webb, Edmund; Oztekin, Alparslan; Zhang, Xiaohui; Cheng, Xuanhong

2017-11-01

Von Willebrand factor (vWF) is a blood glycoprotein that binds with platelets and collagen on injured vessel surfaces to form clots. VWF bioactivity is shear flow induced: at low shear, binding between VWF and other biological entities is suppressed; for high shear rate conditions - as are found near arterial injury sites - VWF elongates, activating its binding with platelets and collagen. Based on parameters derived from single molecule force spectroscopy experiments, we developed a coarse-grain molecular model to simulate bond formation probability as a function of shear rate. By introducing a binding criterion that depends on the conformation of a sub-monomer molecular feature of our model, the model predicts shear-induced binding, even for conditions where binding is highly energetically favorable. We further investigate the influence of various model parameters on the ability to predict shear-induced binding (vWF length, collagen site density and distribution, binding energy landscape, and slip/catch bond length) and demonstrate parameter ranges where the model provides good agreement with existing experimental data. Our results may be important for understanding vWF activity and also for achieving targeted drug therapy via biomimetic synthetic molecules. National Science Foundation (NSF),Division of Mathematical Sciences (DMS).

Condensation induced water hammer safety

International Nuclear Information System (INIS)

Gintner, M.A.

1997-01-01

Condensation induced water hammer events in piping systems can cause catastrophic steam system failures which can result in equipment damage, personal injury, and even death. As an industry, we have learned to become accustomed to the ''banging'' that we often hear in our steam piping systems, and complacent in our actions to prevent it. It is unfortunate that lives are lost needlessly, as this type of water hammer event is preventable if one only applies some basic principles when operating and maintaining their steam systems. At the U. S. Department of Energy's Hanford Site where I work, there was one such accident that occurred in 1993 which took the life of a former co-worker and friend of mine. Hanford was established as part of the Manhattan Project during World War II. it is a 560 square mile complex located along the banks of the Columbia River in Southeastern Washington State. For almost 45 years, hanford's mission was to produce weapons grade plutonium for our nations defense programs. Today, Hanford no longer produces plutonium, but is focused on site clean-up and economic diversification. Hanford still uses steam for heating and processing activities, utilizing over 20 miles of piping distribution systems similar to those found in industry. Although these aging systems are still sound, they cannot stand up to the extreme pressure pulses developed by a condensation induced water hammer

Alkali-labile sites and post-irradiation effects in single-stranded DNA induced by H radicals

International Nuclear Information System (INIS)

Lafleur, M.V.M.; Heuvel, N. van; Woldhuis, J.; Loman, H.

1978-01-01

Single-stranded phiX174 DNA in aqueous solutions has been irradiated in the absence of oxygen, under conditions in which H radicals react with the DNA. It was shown that H radical reactions result in breaks, which contribute approximately 10 per cent inactivation. Further, two types of alkali-labile sites were formed. One was lethal and gave rise to single-strand breaks by alkali and was most probably identical with post-irradiation heat damage and contributed about 33 per cent to the inactivation mentioned above. The other consisted of non-lethal damage, partly dihydropyrimidine derivatives, and was converted to lethal damage by alkali. This followed from experiments in which the DNA was treated with osmium-tetroxide, which oxidized thymine to 5,6-dihydroxydihydrothymine. Treatment with alkali of this DNA gave the same temperature dependence as found for the non-lethal alkali-labile sites in irradiated DNA. A similar temperature dependence was found for dihydrothymine and irradiated pyrimidines with alkali. (author)

Systematic Analysis of Splice-Site-Creating Mutations in Cancer

Directory of Open Access Journals (Sweden)

Reyka G. Jayasinghe

2018-04-01

Full Text Available Summary: For the past decade, cancer genomic studies have focused on mutations leading to splice-site disruption, overlooking those having splice-creating potential. Here, we applied a bioinformatic tool, MiSplice, for the large-scale discovery of splice-site-creating mutations (SCMs across 8,656 TCGA tumors. We report 1,964 originally mis-annotated mutations having clear evidence of creating alternative splice junctions. TP53 and GATA3 have 26 and 18 SCMs, respectively, and ATRX has 5 from lower-grade gliomas. Mutations in 11 genes, including PARP1, BRCA1, and BAP1, were experimentally validated for splice-site-creating function. Notably, we found that neoantigens induced by SCMs are likely several folds more immunogenic compared to missense mutations, exemplified by the recurrent GATA3 SCM. Further, high expression of PD-1 and PD-L1 was observed in tumors with SCMs, suggesting candidates for immune blockade therapy. Our work highlights the importance of integrating DNA and RNA data for understanding the functional and the clinical implications of mutations in human diseases. : Jayasinghe et al. identify nearly 2,000 splice-site-creating mutations (SCMs from over 8,000 tumor samples across 33 cancer types. They provide a more accurate interpretation of previously mis-annotated mutations, highlighting the importance of integrating data types to understand the functional and the clinical implications of splicing mutations in human disease. Keywords: splicing, RNA, mutations of clinical relevance

Siting guidelines and their role in repository site selection

International Nuclear Information System (INIS)

Hanlon, C.L.

1985-01-01

The first requirement of the Nuclear Waste Policy Act was for the Secretary of Energy to issue general guidelines for siting repositories. The guidelines were to specify detailed geologic considerations that would be the primary criteria for the selection of sites in various host rocks, as well as factors that would qualify or disqualify any site from development as a repository. These guidelines were clearly intended to provide not only the framework for the siting program but also the stimulus for establishing effective communication and consultation among the parties involved in the program. The Act further required that the guidelines be a factor in the development of all future decision documents of the Office of Civilian Radioactive Waste Management, including the environmental assessments that would accompany the nomination of sites for characterization, the site-characterization plans that are to be prepared before the sinking of exploratory shafts at any candidate site, and the environmental impact statement that is to support the recommendation of a site for development as a repository. More than two years after its passage, the intention of the Act for the guidelines has been realized. Concurred in by the Nuclear Regulatory Commission on June 22, 1984, and issued by the Department in November 1984, the guidelines include postclosure technical guidelines that apply to conditions governing the long-term performance of the repository system; preclosure technical guidelines that apply to conditions governing the siting, construction, operation, and closure of the repository; and system guidelines whose objective is to ensure that the regulatory requirements of the Environmental Protection Agency and the Nuclear Regulatory Commission are met

Comparative conventional- and quantum dot-labelling strategies for LPS binding site detection in Arabidopsis thaliana mesophyll protoplasts

Directory of Open Access Journals (Sweden)

Londiwe Siphephise Mgcina

2015-05-01

Full Text Available Lipopolysaccharide (LPS from Gram-negative bacteria is recognized as a microbe-associated molecular pattern (MAMP and not only induces an innate immune response in plants, but also stimulates the development of characteristic defense responses. However, identification and characterization of a cell surface LPS-receptor/binding site, as described in mammals, remains elusive in plants. As an amphiphilic, macromolecular lipoglycan, intact LPS potentially contains three MAMP-active regions, represented by the O-polysaccharide chain, the core and the lipid A. Binding site studies with intact labelled LPS were conducted in Arabidopsis thaliana protoplasts and quantified using flow cytometry fluorescence changes. Qdots, which allow non-covalent, hydrophobic labelling were used as a novel strategy in this study and compared to covalent, hydrophilic labelling with Alexa 488. Affinity for LPS-binding sites was clearly demonstrated by concentration-, temperature- and time-dependent increases in protoplast fluorescence following treatment with the labelled LPS. Moreover, this induced fluorescence increase was convincingly reduced following pre-treatment with excess unlabeled LPS, thereby indicating reversibility of LPS binding. Inhibition of the binding process is also reported using endo- and exocytosis inhibitors. Here, we present evidence for the anticipated presence of LPS-specific binding sites in Arabidopsis protoplasts, and furthermore propose Qdots as a more sensitive LPS-labelling strategy in comparison to the conventional Alexa 488 hydrazide label for binding studies.

SiteGuide: An example-based approach to web site development assistance

NARCIS (Netherlands)

Hollink, V.; de Boer, V.; van Someren, M.; Filipe, J.; Cordeiro, J.

2009-01-01

We present ‘SiteGuide’, a tool that helps web designers to decide which information will be included in a new web site and how the information will be organized. SiteGuide takes as input URLs of web sites from the same domain as the site the user wants to create. It automatically searches the pages

Differential active site loop conformations mediate promiscuous activities in the lactonase SsoPox.

Directory of Open Access Journals (Sweden)

Julien Hiblot

Full Text Available Enzymes are proficient catalysts that enable fast rates of Michaelis-complex formation, the chemical step and products release. These different steps may require different conformational states of the active site that have distinct binding properties. Moreover, the conformational flexibility of the active site mediates alternative, promiscuous functions. Here we focused on the lactonase SsoPox from Sulfolobus solfataricus. SsoPox is a native lactonase endowed with promiscuous phosphotriesterase activity. We identified a position in the active site loop (W263 that governs its flexibility, and thereby affects the substrate specificity of the enzyme. We isolated two different sets of substitutions at position 263 that induce two distinct conformational sampling of the active loop and characterized the structural and kinetic effects of these substitutions. These sets of mutations selectively and distinctly mediate the improvement of the promiscuous phosphotriesterase and oxo-lactonase activities of SsoPox by increasing active-site loop flexibility. These observations corroborate the idea that conformational diversity governs enzymatic promiscuity and is a key feature of protein evolvability.

Distinct lipid a moieties contribute to pathogen-induced site-specific vascular inflammation.

Directory of Open Access Journals (Sweden)

Connie Slocum

2014-07-01

Full Text Available Several successful pathogens have evolved mechanisms to evade host defense, resulting in the establishment of persistent and chronic infections. One such pathogen, Porphyromonas gingivalis, induces chronic low-grade inflammation associated with local inflammatory bone loss and systemic inflammation manifested as atherosclerosis. P. gingivalis expresses an atypical lipopolysaccharide (LPS structure containing heterogeneous lipid A species, that exhibit Toll-like receptor-4 (TLR4 agonist or antagonist activity, or are non-activating at TLR4. In this study, we utilized a series of P. gingivalis lipid A mutants to demonstrate that antagonistic lipid A structures enable the pathogen to evade TLR4-mediated bactericidal activity in macrophages resulting in systemic inflammation. Production of antagonistic lipid A was associated with the induction of low levels of TLR4-dependent proinflammatory mediators, failed activation of the inflammasome and increased bacterial survival in macrophages. Oral infection of ApoE(-/- mice with the P. gingivalis strain expressing antagonistic lipid A resulted in vascular inflammation, macrophage accumulation and atherosclerosis progression. In contrast, a P. gingivalis strain producing exclusively agonistic lipid A augmented levels of proinflammatory mediators and activated the inflammasome in a caspase-11-dependent manner, resulting in host cell lysis and decreased bacterial survival. ApoE(-/- mice infected with this strain exhibited diminished vascular inflammation, macrophage accumulation, and atherosclerosis progression. Notably, the ability of P. gingivalis to induce local inflammatory bone loss was independent of lipid A expression, indicative of distinct mechanisms for induction of local versus systemic inflammation by this pathogen. Collectively, our results point to a pivotal role for activation of the non-canonical inflammasome in P. gingivalis infection and demonstrate that P. gingivalis evades immune

The Escherichia coli Tus-Ter replication fork barrier causes site-specific DNA replication perturbation in yeast.

Science.gov (United States)

Larsen, Nicolai B; Sass, Ehud; Suski, Catherine; Mankouri, Hocine W; Hickson, Ian D

2014-04-07

Replication fork (RF) pausing occurs at both 'programmed' sites and non-physiological barriers (for example, DNA adducts). Programmed RF pausing is required for site-specific DNA replication termination in Escherichia coli, and this process requires the binding of the polar terminator protein, Tus, to specific DNA sequences called Ter. Here, we demonstrate that Tus-Ter modules also induce polar RF pausing when engineered into the Saccharomyces cerevisiae genome. This heterologous RF barrier is distinct from a number of previously characterized, protein-mediated, RF pause sites in yeast, as it is neither Tof1-dependent nor counteracted by the Rrm3 helicase. Although the yeast replisome can overcome RF pausing at Tus-Ter modules, this event triggers site-specific homologous recombination that requires the RecQ helicase, Sgs1, for its timely resolution. We propose that Tus-Ter can be utilized as a versatile, site-specific, heterologous DNA replication-perturbing system, with a variety of potential applications.

Feedback from performance assessment to site characterisation. The SITE-94 example

International Nuclear Information System (INIS)

Dverstorp, B.; Geier, J.

1999-01-01

Interaction and information exchange between site characterisation and performance assessment are key features of any successful radioactive waste management programme. Some examples are presented of the types of feedback that can be offered from performance assessment to site characterisation, based on SKI's most recent performance assessment project, SITE-94. SITE-94 in Sweden was an assessment of a hypothetical repository for spent nuclear fuel, based on real data gathered in the surface-based investigation of the Aespoe Hard Rock Laboratory site. Examples of feedback are given concerning quality control of data and site investigation procedures, identification of key parameters for the performance assessment, use of models for planning and evaluation of a site investigation, data sampling strategies, and guidance on future priorities for further development of site investigation methods. Because site characterisation serves multiple purposes, including provision of data for repository design and construction, it must account for and compromise among requirements from several lines of analyses in the performance assessment. (author)

Effect of morphine-induced antinociception is altered by AF64A-induced lesions on cholinergic neurons in rat nucleus raphe magnus.

Science.gov (United States)

Abe, Kenji; Ishida, Kota; Kato, Masatoshi; Shigenaga, Toshiro; Taguchi, Kyoji; Miyatake, Tadashi

2002-11-01

To examine the role of cholinergic neurons in the nucleus raphe magnus (NRM) in noxious heat stimulation and in the effects of morphine-induced antinociception by rats. After the cholinergic neuron selective toxin, AF64A, was microinjected into the NRM, we examined changes in the antinociceptive threshold and effects of morphine (5 mg/kg, ip) using the hot-plate (HP) and tail-flick (TF) tests. Systemic administration of morphine inhibited HP and TF responses in control rats. Microinjection of AF64A (2 nmol/site) into the NRM significantly decreased the threshold of HP response after 14 d, whereas the TF response was not affected. Morphine-induced antinociception was significantly attenuated in rats administered AF64A. Extracellular acetylcholine was attenuated after 14 d to below detectable levels in rats given AF64A. Naloxone (1 microg/site) microinjected into control rat NRM also antagonized the antinociceptive effect of systemic morphine. These findings suggest that cholinergic neuron activation in the NRM modulates the antinociceptive effect of morphine simultaneously with the opiate system.

Soil organic matter status in forest soils - possible indicators for climate change induced site shifts

Science.gov (United States)

Koch, Nadine; Thiele-Bruhn, Sören

2010-05-01

The quantity and quality of soil organic matter (SOM) and SOM pools and thus the soil properties related to carbon sequestration and water retention are not constant but exhibit considerable variation through changing climate. In total changes in soil fertility and an increase in plant stress are expected. This is relevant for northwest Europe as well and may have economic and social impacts since functions of forests for wood production, groundwater recharge, soil protection and recreation might be affected. The study is done by comparative investigation of selected sites at four watersheds that represent typical forest stands in the region of Luxembourg and South West Germany. The aim is to identify SOM storage and stability in forest soils and its dependence on site properties and interaction with tree stand conditions. According to state of the art fractionation schemes functional C pools in forest soils and their stabilization mechanisms are investigated. In particular, distribution patterns are determined depending on location, tree stand and climatic conditions. Aim is to identify characteristics of SOM stability through fractionation of SOM according to density, particle size and chemical extractability and their subsequent analytical characterization. So far, reasons about the origin, composition and stabilization mechanisms underlying the different SOM pools are not fully understood. Presented are different patterns of distribution of SOM in relation to land use and site conditions, as well as similarities and differences between the different forest soils and results in addition to passive OM pool, which is mainly responsible for long-term stabilization of carbon in soils. These are aligned with selected general' soil properties such as pH, CEC and texture.

Nonequivalence of alpha-bungarotoxin binding sites in the native nicotinic receptor molecule

International Nuclear Information System (INIS)

Conti-Tronconi, B.M.; Tang, F.; Walgrave, S.; Gallagher, W.

1990-01-01

In the native, membrane-bound form of the nicotinic acetylcholine receptor (M-AcChR) the two sites for the cholinergic antagonist alpha-bungarotoxin (alpha-BGT) have different binding properties. One site has high affinity, and the M-AcChR/alpha-BGT complexes thus formed dissociate very slowly, similar to the complexes formed with detergent-solubilized AcChR (S-AcChR). The second site has much lower affinity (KD approximately 59 +/- 35 nM) and forms quickly reversible complexes. The nondenaturing detergent Triton X-100 is known to solubilize the AcChR in a form unable, upon binding of cholinergic ligands, to open the ion channel and to become desensitized. Solubilization of the AcChR in Triton X-100 affects the binding properties of this second site and converts it to a high-affinity, slowly reversible site. Prolonged incubation of M-AcChR at 4 degrees C converts the low-affinity site to a high-affinity site similar to those observed in the presence of Triton X-100. Although the two sites have similar properties when the AcChR is solubilized in Triton X-100, their nonequivalence can be demonstrated by the effect on alpha-BGT binding of concanavalin A, which strongly reduces the association rate of one site only. The Bmax of alpha-BGT to either Triton-solubilized AcChR or M-AcChR is not affected by the presence of concanavalin A. Occupancy of the high-affinity, slowly reversible site in M-AcChR inhibits the Triton X-100 induced conversion to irreversibility of the second site. At difference with alpha-BGT, the long alpha-neurotoxin from Naja naja siamensis venom (alpha-NTX) binds with high affinity and in a very slowly reversible fashion to two sites in the M-AcChR. We confirm here that Triton-solubilized AcChR or M-AcChR binds in a very slowly reversible fashion the same amount of alpha-NTX

Impact of MicroRNA Levels, Target-Site Complementarity, and Cooperativity on Competing Endogenous RNA-Regulated Gene Expression.

Science.gov (United States)

Denzler, Rémy; McGeary, Sean E; Title, Alexandra C; Agarwal, Vikram; Bartel, David P; Stoffel, Markus

2016-11-03

Expression changes of competing endogenous RNAs (ceRNAs) have been proposed to influence microRNA (miRNA) activity and thereby regulate other transcripts containing miRNA-binding sites. Here, we find that although miRNA levels define the extent of repression, they have little effect on the magnitude of the ceRNA expression change required to observe derepression. Canonical 6-nt sites, which typically mediate modest repression, can nonetheless compete for miRNA binding, with potency ∼20% of that observed for canonical 8-nt sites. In aggregate, low-affinity/background sites also contribute to competition. Sites with extensive additional complementarity can appear as more potent, but only because they induce miRNA degradation. Cooperative binding of proximal sites for the same or different miRNAs does increase potency. These results provide quantitative insights into the stoichiometric relationship between miRNAs and target abundance, target-site spacing, and affinity requirements for ceRNA-mediated gene regulation, and the unusual circumstances in which ceRNA-mediated gene regulation might be observed. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Permeability Estimation Directly From Logging-While-Drilling Induced Polarization Data

DEFF Research Database (Denmark)

Fiandaca, G.; Maurya, P.K.; Balbarini, Nicola

2018-01-01

In this study we present the prediction of permeability from time‐domain spectral induced polarization (IP) data, measured in boreholes on undisturbed formations using the El‐log logging‐while‐drilling technique. We collected El‐log data and hydraulic properties on unconsolidated Quaternary...... and Miocene deposits in boreholes at three locations at a field site in Denmark, characterized by different electrical water conductivity and chemistry. The high vertical resolution of the El‐log technique matches the lithological variability at the site, minimizing ambiguity in the interpretation originating...

Site characterization plan:

International Nuclear Information System (INIS)

1988-01-01

The Yucca Mountain site in Nevada is one of three candidate sites for the first geologic repository for radioactive waste. On May 28, 1986, it was recommended for detailed study in a program of site characterization. This site characterization plan (SCP) has been prepared in accordance with the requirements of the Nuclear Waste Policy Act to summarize the information collected to date about the geologic conditions at the site;to describe the conceptual designs for the repository and the waste package;and to present the plans for obtaining the geologic information necessary to demonstrate the suitability of the site for repository, to design the repository and the waste package, to prepare an environmental impact statement, and to obtain from the US Nuclear Regulatory Commission (NRC) an authorization to construct the repository. This introduction begins with a brief section on the process for siting and developing a repository, followed by a discussion of the pertinent legislation and regulations. A description of site characterization is presented next;it describes the facilities to be constructed for the site characterization program and explains the principal activities to be conducted during the program. Finally, the purpose, content, organizing principles, and organization of this site characterization plan are outlined, and compliance with applicable regulations is discussed

A New Aedes (Finlaya) of the Niveus-Subgroup (Diptera: Culicidae)

Science.gov (United States)

1987-01-01

specimens. bRange (Mode). TNTC-Too numerous to count. Table 3. Number of branches for setae of the fourth-instar larva of Aedes (Finlaya...2) 1,2(l) 1,2(2) 1,2(l) 1 2-4(2) 2,3(2) 7-ll(7) 1 1 1,2(l) aBased on counts from at least 10 setae on the holotype and 8 other specimens. bRange

Preliminary analysis of the potential for thermally-induced rock fracture around high-level waste containers

International Nuclear Information System (INIS)

Ratigan, J.L.

1976-01-01

The major results are: the development of parametric formulations relating the potential for thermally induced fracturing in the high-level radioactive waste repository concept to the elastic and thermal properties of the site rock and the depth of the excavation, and the recognition of a need to determine the actual ''failure envelope'' for any potential site rock in the laboratory and adjust the parametric relations appropriately. Analysis of five rock types indicated that none would experience elastic/brittle failure due to the thermal stresses induced by the introduction of a 5 kW heat source. However, the rock strengths and elastic properties are laboratory values and not in situ values

Future vegetation types and related main processes for Olkiluoto site

International Nuclear Information System (INIS)

Haapanen, R.

2007-07-01

This working report summarizes current knowledge of the land up-lift induced vegetation succession and future vegetation types on Olkiluoto Island and its surroundings. The report is based on generic literature and site-specific studies concerning Olkiluoto Island. Current vegetation on Olkiluoto Island and typical succession lines on different soil types are described, as well as main factors affecting the succession. Most relevant materials on hand are listed. Some problems and possible areas to be emphasized before using the data in modelling work are pointed out. (orig.)

Analysis of elemental composition of porcelains unearthed from Waguantan kiln site by PIXE-RBS

Science.gov (United States)

Zhou, Z.; Zhang, K.; Xia, C. D.; Liu, M. T.; Zhu, J. J.; An, Z.; Bai, B.

2015-03-01

A method combining proton-induced X-ray emission spectrometry (PIXE) and Rutherford backscattering spectrometry (RBS) was used to determine the composition of 61 porcelain shards from the Yuan Dynasty (1271-1368 A.D.) unearthed from the Waguantan kiln site at Tianzhu County in Guizhou Province, China. Based on our previous experimental setup, an electron gun device with a LaB6 crystal cathode was installed to solve the problem created when the incident proton beams generated electric charge accumulations on the surfaces of the insulating porcelain samples, which induced a large bremsstrahlung background. The use of the electron gun has largely eliminated the large bremsstrahlung background and has therefore improved the detection limits for elements, especially for trace elements, and made it possible to determine the origin of the porcelains based on the trace elements. Major and trace elemental compositions of the porcelain bodies and glazes measured by PIXE and RBS were analyzed by the factor analysis method. The factor analysis showed that a few pieces of porcelain with a style similar to the porcelain of the Longquan kiln among the unearthed porcelains from the Waguantan kiln site did not have obvious differences in elemental compositions from other remaining porcelains unearthed from the Waguantan kiln site, indicating that the pieces of unearthed porcelain with the Longquan kiln style did in fact belong to the product fired locally by imitating the model of the Longquan celadon with local raw materials. This result therefore indicated that the Longquan kiln technology that originated from the Five Dynasties (907-960 A.D.) had been propagated to the Waguantan kiln site of Guizhou Province in the Yuan Dynasty.

US -endorphin-(1-27) is a naturally occurring antagonist to etorphine-induced analgesia

Energy Technology Data Exchange (ETDEWEB)

Nicolas, P.; Li, C.H.

1985-05-01

The potent opioid peptide US -endorphin is found in the brain and pituitary with two related fragments, US -endorphin-(1-27) and US -endorphin-(1-26). The fragments, retain substantial opioid-receptor binding activity but are virtually inactive analgesically. US -Endorphin-(1-27) inhibits US -endorphin-induced and etorphine-induced analgesia when coinjected intracerebroventricularly into mice. Antagonism by competition at the same site(s) is suggested from parallel shifts of the dose-response curves of etorphine or US -endorphin in the presence of US -endorphin-(1-27). Its potency is 4-5 times greater than that of the opiate antagonist naloxone. US -Endorphin-(1-26) does not antagonize the antinociceptive action of etorphine or US -endorphin in doses up to 500 pmol per animal.

SiteFind: A software tool for introducing a restriction site as a marker for successful site-directed mutagenesis

Directory of Open Access Journals (Sweden)

Evans Paul M

2005-12-01

Full Text Available Abstract Background Site-directed mutagenesis is a widely-used technique for introducing mutations into a particular DNA sequence, often with the goal of creating a point mutation in the corresponding amino acid sequence but otherwise leaving the overall sequence undisturbed. However, this method provides no means for verifying its success other than sequencing the putative mutant construct: This can quickly become an expensive method for screening for successful mutations. An alternative to sequencing is to simultaneously introduce a restriction site near the point mutation in manner such that the restriction site has no effect on the translated amino acid sequence. Thus, the novel restriction site can be used as a marker for successful mutation which can be quickly and easily assessed. However, finding a restriction site that does not disturb the corresponding amino acid sequence is a time-consuming task even for experienced researchers. A fast and easy to use computer program is needed for this task. Results We wrote a computer program, called SiteFind, to help us design a restriction site within the mutation primers without changing the peptide sequence. Because of the redundancy of genetic code, a given peptide can be encoded by many different DNA sequences. Since the list of possible restriction sites for a given DNA sequence is not always obvious, SiteFind automates this task. The number of possible sequences a computer program must search through increases exponentially as the sequence length increases. SiteFind uses a novel "moving window" algorithm to reduce the number of possible sequences to be searched to a manageable level. The user enters a nucleotide sequence, specifies what amino acid residues should be changed in the mutation, and SiteFind generates a list of possible restriction sites and what nucleotides must be changed to introduce that site. As a demonstration of its use, we successfully generated a single point mutation

Hair Barrette Induced Cochlear Implant Receiver Stimulator Site Infection with Extrusion

Directory of Open Access Journals (Sweden)

Trung N. Le

2015-01-01

Full Text Available Background. Cochlear implant infections and extrusion are uncommon but potentially devastating complications. Recent literature suggests conservative management can be employed. Local measures inclusive of aggressive surgical debridement with vascularized flaps and parenteral antibiotics represent a viable option and often permit device salvage. However, explantation should be considered if there is evidence of systemic, intracranial, or intractable infection. Method. A Case report and literature review. Case Report. This case illustrates a complicated local wound infection associated with cochlear implantation due to transcutaneous adherence of a ferrous hair barrette to a cochlear implant magnet. Reconstruction of computed tomography (CT data with 3D volume rendering significantly improved the value of the images and facilitated patient counseling as well as operative planning. Conclusion. Cochlear implant infections can be associated with foreign bodies. CT images are beneficial in the evaluation of cochlear implant complications. 3D CT images provide a comprehensive view of the site of interest, displaying the relationship of the hardware to the skull and soft tissues, while minimizing associated artifacts. Cochlear implant patients should consider use of nonmetallic hair devices.

Copper(II) Binding Sites in N-Terminally Acetylated α-Synuclein: A Theoretical Rationalization.

Science.gov (United States)

Ramis, Rafael; Ortega-Castro, Joaquín; Vilanova, Bartolomé; Adrover, Miquel; Frau, Juan

2017-08-03

The interactions between N-terminally acetylated α-synuclein and Cu(II) at several binding sites have been studied with DFT calculations, specifically with the M06 hybrid functional and the ωB97X-D DFT-D functional. In previous experimental studies, Cu(II) was shown to bind several α-synuclein residues, including Met1-Asp2 and His50, forming square planar coordination complexes. Also, it was determined that a low-affinity binding site exists in the C-terminal domain, centered on Asp121. However, in the N-terminally acetylated protein, present in vivo, the Met1 site is blocked. In this work, we simplify the representation of the protein by modeling each experimentally found binding site as a complex between an N-terminally acetylated α-synuclein dipeptide (or several independent residues) and a Cu(II) cation, and compare the results with a number of additional, structurally analogous sites not experimentally found. This way of representing the binding sites, although extremely simple, allows us to reproduce experimental results and to provide a theoretical rationale to explain the preference of Cu(II) for certain sites, as well as explicit geometrical structures for the complexes formed. These results are important to understand the interactions between α-synuclein and Cu(II), one of the factors inducing structural changes in the protein and leading to aggregated forms of it which may play a role in neurodegeneration.

The Open Form Inducer Approach for Structure-Based Drug Design.

Directory of Open Access Journals (Sweden)

Daniel Ken Inaoka

Full Text Available Many open form (OF structures of drug targets were obtained a posteriori by analysis of co-crystals with inhibitors. Therefore, obtaining the OF structure of a drug target a priori will accelerate development of potent inhibitors. In addition to its small active site, Trypanosoma cruzi dihydroorotate dehydrogenase (TcDHODH is fully functional in its monomeric form, making drug design approaches targeting the active site and protein-protein interactions unrealistic. Therefore, a novel a priori approach was developed to determination the TcDHODH active site in OF. This approach consists of generating an "OF inducer" (predicted in silico to bind the target and cause steric repulsion with flexible regions proximal to the active site that force it open. We provide the first proof-of-concept of this approach by predicting and crystallizing TcDHODH in complex with an OF inducer, thereby obtaining the OF a priori with its subsequent use in designing potent and selective inhibitors. Fourteen co-crystal structures of TcDHODH with the designed inhibitors are presented herein. This approach has potential to encourage drug design against diseases where the molecular targets are such difficult proteins possessing small AS volume. This approach can be extended to study open/close conformation of proteins in general, the identification of allosteric pockets and inhibitors for other drug targets where conventional drug design approaches are not applicable, as well as the effective exploitation of the increasing number of protein structures deposited in Protein Data Bank.

Remaining Sites Verification Package for the 128-B-3 Burn Pit Site, Waste Site Reclassification Form 2006-058

Energy Technology Data Exchange (ETDEWEB)

L. M. Dittmer

2006-11-17

The 128-B-3 waste site is a former burn and disposal site for the 100-B/C Area, located adjacent to the Columbia River. The 128-B-3 waste site has been remediated to meet the remedial action objectives specified in the Remaining Sites ROD. The results of verification sampling demonstrated that residual contaminant concentrations do not preclude any future uses and allow for unrestricted use of shallow zone soils. The results of sampling at upland areas of the site also showed that residual contaminant concentrations are protective of groundwater and the Columbia River.

Site characterization plan:

International Nuclear Information System (INIS)

1988-01-01

The Yucca Mountain site in Nevada is one of three candidate sites for the first geologic repository for radioactive waste. On May 28, 1986, it was recommended for detailed study in a program of site characterization. This site characterization plan (SCP) has been prepared in acordance with the requirements of the Nuclear Waste Policy Act to summarize the information collected to date about the geologic conditions at the site;to describe the conceptual designs for the repository and the waste package and to present the plans for obtaining the geologic information necessary to demonstrate the suitability of the site for a repository, to design the repository and the waste package, to prepare an environmental impact statement, and to obtain from the US Nuclear Regulatory Commission (NRC) an authorization to construct the repository. This introduction begins with a brief section on the process for siting and eveloping a repository, followed by a discussion of the pertinent legislation and regulations. A description of site characterization is presented next;it describes the facilities to be constructed for the site characterization program and explains the principal activities to be conducted during the program. Finally, the purpose, content, organizing prinicples, and organization of this site characterization plan are outlined, and compliance with applicable regulations is discussed. 880 refs., 130 figs., 25 tabs

On multi-site damage identification using single-site training data

Science.gov (United States)

Barthorpe, R. J.; Manson, G.; Worden, K.

2017-11-01

This paper proposes a methodology for developing multi-site damage location systems for engineering structures that can be trained using single-site damaged state data only. The methodology involves training a sequence of binary classifiers based upon single-site damage data and combining the developed classifiers into a robust multi-class damage locator. In this way, the multi-site damage identification problem may be decomposed into a sequence of binary decisions. In this paper Support Vector Classifiers are adopted as the means of making these binary decisions. The proposed methodology represents an advancement on the state of the art in the field of multi-site damage identification which require either: (1) full damaged state data from single- and multi-site damage cases or (2) the development of a physics-based model to make multi-site model predictions. The potential benefit of the proposed methodology is that a significantly reduced number of recorded damage states may be required in order to train a multi-site damage locator without recourse to physics-based model predictions. In this paper it is first demonstrated that Support Vector Classification represents an appropriate approach to the multi-site damage location problem, with methods for combining binary classifiers discussed. Next, the proposed methodology is demonstrated and evaluated through application to a real engineering structure - a Piper Tomahawk trainer aircraft wing - with its performance compared to classifiers trained using the full damaged-state dataset.

COSMID: A Web-based Tool for Identifying and Validating CRISPR/Cas Off-target Sites

Directory of Open Access Journals (Sweden)

Thomas J Cradick

2014-01-01

Full Text Available Precise genome editing using engineered nucleases can significantly facilitate biological studies and disease treatment. In particular, clustered regularly interspaced short palindromic repeats (CRISPR with CRISPR-associated (Cas proteins are a potentially powerful tool for modifying a genome by targeted cleavage of DNA sequences complementary to designed guide strand RNAs. Although CRISPR/Cas systems can have on-target cleavage rates close to the transfection rates, they may also have relatively high off-target cleavage at similar genomic sites that contain one or more base pair mismatches, and insertions or deletions relative to the guide strand. We have developed a bioinformatics-based tool, COSMID (CRISPR Off-target Sites with Mismatches, Insertions, and Deletions that searches genomes for potential off-target sites (http://crispr.bme.gatech.edu. Based on the user-supplied guide strand and input parameters, COSMID identifies potential off-target sites with the specified number of mismatched bases and insertions or deletions when compared with the guide strand. For each site, amplification primers optimal for the chosen application are also given as output. This ranked-list of potential off-target sites assists the choice and evaluation of intended target sites, thus helping the design of CRISPR/Cas systems with minimal off-target effects, as well as the identification and quantification of CRISPR/Cas induced off-target cleavage in cells.

Nevada Test Site Environmental Report 2005, Attachment A - Site Description

International Nuclear Information System (INIS)

Cathy A. Wills

2006-01-01

This appendix to the ''Nevada Test Site Environmental Report 2005'', dated October 2006 (DOE/NV/11718--1214; DOE/NV/25946--007) expands on the general description of the Nevada Test Site (NTS) presented in the Introduction. Included are subsections that summarize the site?s geological, hydrological, climatological, and ecological setting. The cultural resources of the NTS are also presented. The subsections are meant to aid the reader in understanding the complex physical and biological environment of the NTS. An adequate knowledge of the site's environment is necessary to assess the environmental impacts of new projects, design and implement environmental monitoring activities for current site operations, and assess the impacts of site operations on the public residing in the vicinity of the NTS. The NTS environment contributes to several key features of the site which afford protection to the inhabitants of adjacent areas from potential exposure to radioactivity or other contaminants resulting from NTS operations. These key features include the general remote location of the NTS, restricted access, extended wind transport times, the great depths to slow-moving groundwater, little or no surface water, and low population density. This appendix complements the annual summary of monitoring program activities and dose assessments presented in the main body of this report

Nevada Test Site Environmental Report 2005, Attachment A - Site Description

Energy Technology Data Exchange (ETDEWEB)

Cathy A. Wills

2006-10-01

This appendix to the ''Nevada Test Site Environmental Report 2005'', dated October 2006 (DOE/NV/11718--1214; DOE/NV/25946--007) expands on the general description of the Nevada Test Site (NTS) presented in the Introduction. Included are subsections that summarize the site?s geological, hydrological, climatological, and ecological setting. The cultural resources of the NTS are also presented. The subsections are meant to aid the reader in understanding the complex physical and biological environment of the NTS. An adequate knowledge of the site's environment is necessary to assess the environmental impacts of new projects, design and implement environmental monitoring activities for current site operations, and assess the impacts of site operations on the public residing in the vicinity of the NTS. The NTS environment contributes to several key features of the site which afford protection to the inhabitants of adjacent areas from potential exposure to radioactivity or other contaminants resulting from NTS operations. These key features include the general remote location of the NTS, restricted access, extended wind transport times, the great depths to slow-moving groundwater, little or no surface water, and low population density. This appendix complements the annual summary of monitoring program activities and dose assessments presented in the main body of this report.

Siting Practices and Site Licensing Process for New Reactors in Canada

International Nuclear Information System (INIS)

Vos, Marcel de

2011-01-01

'Siting' in Canada is composed of Site Evaluation and Site Selection. As outlined in CNSC Regulatory Document RD-346 Site Evaluation for New Nuclear Power Plants (based on IAEA NS-R-3), prior to the triggering of the Environmental Assessment (EA) and licensing processes, the proponent is expected to use a robust process to characterize proposed sites over the full life cycle of the facility, and then develop a fully documented defense of the site selection case. This case forms the backbone for submissions in support of the EA and the application for a License to Prepare Site which will be reviewed by the CNSC and other applicable federal authorities. The Environmental Assessment process and License to Prepare Site in Canada do not require a proponent to select a specific design; however, CNSC does not accept a 'black box' approach to siting. CNSC balances the level of design information required with the extent of safety assurance desired for any designs being contemplated for the proposed site. Nevertheless, the design information submitted must be sufficient to justify the site as suitable for all future licensing stages. The depth of plant design information contributes significantly to the credibility of the applicant's case for both the EA and application for License to Prepare Site. The review process utilizes an assessment plan with defined review stages and timelines. The outcome of these reviews is a series of recommendations to a federal government appointed Joint Review Panel (which also serves as a panel of the 'Commission') which, following public hearings, renders a decision regarding the EA, and subsequently, the application for a License to Prepare Site. (author)

The Mn site in Mn-doped GaAs nanowires: an EXAFS study

International Nuclear Information System (INIS)

D’Acapito, F; Rovezzi, M; Boscherini, F; Jabeen, F; Bais, G; Piccin, M; Rubini, S; Martelli, F

2012-01-01

We present an EXAFS study of the Mn atomic environment in Mn-doped GaAs nanowires. Mn doping has been obtained either via the diffusion of the Mn used as seed for the nanowire growth or by providing Mn during the growth of Au-induced wires. As a general finding, we observe that Mn forms chemical bonds with As but is not incorporated in a substitutional site. In Mn-induced GaAs wires, Mn is mostly found bonded to As in a rather disordered environment and with a stretched bond length, reminiscent of that exhibited by MnAs phases. In Au-seeded nanowires, along with stretched MnAs coordination, we have found the presence of Mn in a MnAu intermetallic compound. (paper)

Different critical perinatal periods and hypothalamic sites of oestradiol action in the defeminisation of luteinising hormone surge and lordosis capacity in the rat.

Science.gov (United States)

Sakakibara, M; Deura, C; Minabe, S; Iwata, Y; Uenoyama, Y; Maeda, K-I; Tsukamura, H

2013-03-01

Female rats show a gonadotrophin-releasing hormone (GnRH)/luteinising hormone (LH) surge in the presence of a preovulatory level of oestrogen, whereas males do not because of brain defeminisation during the developmental period by perinatal oestrogen converted from androgen. The present study aimed to identify the site(s) of oestrogen action and the critical period for defeminising the mechanism regulating the GnRH/LH surge. Animals given perinatal treatments, such as steroidal manipulations, brain local implantation of oestradiol (E(2) ) or administration of an NMDA antagonist, were examined for their ability to show an E(2) -induced LH surge at adulthood. Lordosis behaviour was examined to compare the mechanisms defeminising the GnRH/LH surge and sexual behaviour. A single s.c. oestradiol-benzoate administration on either the day before birth (E21), the day of birth (D0) or day 5 (D5) postpartum completely abolished the E(2) -induced LH surge at adulthood in female rats, although the same treatment did not inhibit lordosis. Perinatal castration on E21 or D0 partially rescued the E2-induced LH surge in genetically male rats, whereas castration from E21 to D5 totally rescued lordosis. Neonatal E(2) implantation in the anterior hypothalamus including the anteroventral periventricular nucleus (AVPV)/preoptic area (POA) abolished the E(2) -induced LH surge in female rats, whereas E(2) implantation in the mid and posterior hypothalamic regions had no inhibitory effect on the LH surge. Lordosis was not affected by neonatal E(2) implantation in any hypothalamic regions. In male rats, neonatal NMDA antagonist treatment rescued lordosis but not the LH surge. Taken together, these results suggest that an anterior hypothalamic region such as the AVPV/POA region is a perinatal site of oestrogen action where the GnRH/LH regulating system is defeminised to abolish the oestrogen-induced surge. The mechanism for defeminisation of the GnRH/LH surge system might be different from

SITE-94. Site specific base data for the performance assessment

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